Objective: Tongue squamous cell carcinoma (TSCC) is a prominent type of oral cancer. Despite the numerous research studies on SCC and microRNAs (miRs), the relation between TSCC and miR-135b-5p is poorly discussed. This experiment aims to find out the possible effect of miR-135b-5p on TSCC with the network of its downstream genes.
Study Design: TSCC tissues and adjacent normal tissues were harvested. Then, expression of miR-135b-5p and AT-rich interactive domain‑containing protein 1A gene (ARID1A) and the phosphatidyl inositol 3-kinase/protein kinase B (PI3K/AKT) pathway was analyzed. After the transfection of miR-135b-5p inhibitor and its negative control into TSCC cells, functional assays were employed to measure cell proliferation, apoptosis, and cycle. Next, the target relation between miR-135b-5p and ARID1A was confirmed. In addition, the fact that miR-135b-5p promoted TSCC development via mediating ARID1A was demonstrated by functional rescue experiment.
Results: miR-135b-5p was upregulated in TSCC tissues and cells, while ARID1A was suppressed (p< 0.05). Silenced miR-135b-5p discouraged TSCC cell proliferation, improved apoptosis, induced cell cycle arrest, and increased ARID1A expression while inactivating the PI3K/AKT axis (p<0.05). Furthermore, knockdown of ARID1A reversed the impacts on TSCC cell proliferation and apoptosis exerted by silencing miR-135b-5p.
Conclusion: This research supported that silenced miR-135b-5p impeded TSCC proliferation and apoptosis by promoting ARID1A and inactivating the PI3K/AKT axis, which may provide some indications for TSCC alleviation.
Keywords: apoptosis; ARID1A; ARID1A protein, human; carcinoma, squamous cell; cell line, tumor; cell proliferation; drug resistance, neoplasm; microRNA-135b-5p; microRNAs; PI3K/AKT pathway; neoplasm metastasis; neoplastic stem cells; proliferation; protein binding; tongue; tongue squamous cell carcinoma
BACKGROUND: Sequential Epstein-Barr virus (EBV)–positive B cell lymphoma to the initial diagnosis of angioimmunoblastic T cell lymphoma (AITL) is very rare, the exact mechanism and standard therapy of which is still being explored. CASE: A 50-year-old man was admitted to our hospital in January 2014 with a three-week history of enlargement of multiple lymph nodes. His initial pathological evaluation indicated AILT. The reactivation of EBV was observed during the immunosuppression therapy for AITL, accompanied by onset of subcutaneous nodules proven to be EBV-positive diffuse large B cell lymphoma (DLBCL) based on the pathological findings of rebiopsy. The patient was successfully treated with chidamide, a histone deacetylase (HDAC) inhibitor, and rituximab.
Conclusion: The sufficient surveillance for serum EBV and repeat biopsy is necessary for patients with AITL, and this treatment modality may become an active option.
Keywords: angioimmunoblastic T cell lymphoma, Epstein-Barr virus, HDAC inhibitor, non-Hodgkin lymphoma, peripheral T cell lymphoma
Objective: To probe into the influence of miR-21 on the proliferation as well as apoptosis of oral squamous cell carcinoma (OSCC) and its causative role.
Study Design: We adopted microarray for detecting the differentially expressed genes in OSCC tumor tis-sues and paracancerous tissues. We assessed the link of miR-21 expression with tumor size, lymph node metastasis, and tumor differentiation. We employed CCK-8 and EdU assay for detecting the impact of miR-21 inhibitor and miR-21 mimic on Cal-27 cell proliferation, as well as TUNEL and AnnexinV-FITC/PI double staining for detecting miR-21 expression on cell apoptosis. We forecasted the possible target of miR-21 via TargetScan, as well as detected the interaction of miR-21 with PTEN via luciferase reporter experiment. The function of miR-21 expression in PTEN signaling pathway was monitored via western blot. We constructed PTEN overexpression plasmid and conducted rescue experiment to evaluate overexpressed PTEN on miR-21–induced proliferation.
Results: Microarray and RT-qPCR indicated that miR-21 expression increased demonstrably in OSCC. Subsequently, statistical analysis showed that miR-21 expression was plainly correlated with tumor size, lymph node metastasis, tumor differentiation, and smoking history. CCK-8 and EdU method exhibited that miR-21 mimics manifestly promoted Cal-27 cell proliferation, while miR-21 inhibitor blatantly inhibited Cal-27 cell proliferation. TUNEL and V-FITC/PI double staining assay showed that miR-21 inhibitor conspicuously promoted Cal-27 cell apoptosis. CCK-8 and EdU assay exhibited that overexpressed PTEN abolished the pro-proliferation influence of miR-21 mimic. TUNEL and V-FITC/PI experiments pointed out that knocking down PTEN abrogated the pro-apoptosis impact of miR-21 inhibitor.
Conclusion: miR-21 contributes to OSCC cell proliferation via targeting PTEN and inhibits its apoptosis.
Keywords: Akt/PKB signaling pathway; apoptosis; biomarkers, tumor; carcinoma, squamous cell; cell line, tumor; cell proliferation; microRNAs; miR-21; miRNA-21; mouth neoplasms; oral cancer; oral squamous cell carcinoma; proliferation; real time PCR
Objective: To analyze the sonographic features of different histopathological subtypes of borderline ovarian tumors (BOTs) confirmed by pathology, and to study the ultrasound performances of various types in borderline ovarian tumors.
Study Design: Retrospective analysis was performed on the pathological results and ultrasound projection findings of 129 patients diagnosed as BOTs by ultrasound department of our hospital from January 2012 to November 2019. All patients were confirmed by surgical pathology and scanned consecutively by the investigators using transabdominal or transvaginal ultrasound examination.
Results: Serous borderline tumors (SBOTs) were observed, and the prevalence rate (53%) was significantly higher than that of other subtypes, and the probability of bilateral lesions was higher (40%). The sonogram often showed ultrasound features of papillary neoplasm in the lesion and good internal echo (p<0.05). Mucinous borderline ovarian tumors (MBOTs) were mostly unilateral lesions (86%). The prevalence was second only to SBOTs. Histomorphological examinations were divided into gastrointestinal-type and endocervical-type. Among them, the gastrointestinal type of MBOTs were mostly unilateral, and their incidence was higher than that of endocervical-type of MBOTs. Compared with other pathological subtypes, the gastrointestinal type is more likely to show the sonographic characteristics of huge space occupying in the pelvic and abdominal cavity (mean diameter >10 cm), polycystic, multiple septums, and poor internal echo (p<0.05). The ultrasonographic features of the endocervical-type of MBOTs were similar to those of SBOTs. Compared with gastrointestinal type, the sonographic images showed smaller lesion diameter, less septal or cyst, and more papillary excrescences in the tumor (p<0.05). The borderline clear cell tumor is the intermediate transition between the clear cell adenofibroma and the clear cell carcinoma. The clinical manifestations are diverse and lack specificity. The histology of sonography was mainly solid, and the multiple microcapsules were honeycomb-like. It can also be shown as cystic. Among the 169 patients with BOTs, 20 cases of SBOTs, 17 cases of MBOTs, and 10 cases of other rare subtypes were complicated with other diseases or multiple subtypes. This study did not find significant ultrasonic characteristics were used for distinguish them from other subtypes.
Conclusion: BOTs is a common disease in women during the reproductive period. It is characterized by the development of malignant tumors. Its clinical and pathological subtypes are complex and diverse. It leads many doctors to use the terms “large pelvic mass” and “solid ovarian mass” for diagnosis because of their lack of experience and understanding.
Keywords: adenocarcinoma, mucinous; adenocarcinoma, serous; borderline ovarian tumors; diagnostic imaging; ovarian neoplasms; papillary neoplasms; prognosis; transvaginal ultrasound, ultrasonography
Objective: The association between telomerase reverse transcriptase (TERT) promoter mutation and outcome of melanoma is unclear and controversial. We aim to conduct a meta-analysis and investigate whether the TERT promoter mutation is a prognostic factor of melanoma.
Study Design: Appropriate studies were searched in 3 databases: PubMed, Web of Science, and Embase. Pooled hazard ratios (HRs) were counted through random effects model.
Results: Heterogeneity was moderate in overall survival (OS) (I2=43.7%, p=0.059) and low in disease-free survival (DFS) (I2=0.0%, p=0.587). Sensitivity analysis indicated that the removal of any of the study did not affect the final results. Evidence for publication bias was not found (Begg’s test, p=0.281; Egger’s test, p=0.078). The pooled OS HRs from combined effects analysis was determined (HR 1.07; 95% CI 0.83–1.39, p=0.585), together with the pooled HRs of DFS (HR 1.65; 95% CI 1.02–2.66, p=0.042). TERT promoter mutation predicted a good outcome in meta-static melanoma patients (HR 0.66; 95% CI 0.46–0.96, p=0.042). The pooled HRs of combined mutation in TERT promoter and BRAF (HR 6.27; 95% CI 2.7–14.58, p=0.000) predicted a bad outcome in melanoma patients.
Conclusion: TERT promoter mutation significantly predicted poor DFS outcome but, on the contrary, predicted a good outcome in metastatic melanoma patients. The combined TERT promoter and BRAF mutation was a significant independent factor of OS in melanoma patients.
Keywords: melanoma; meta-analysis; mutation; prognosis; promoter regions, genetic; skin neoplasms; telomerase; TERT promoter mutation; TERT protein, human
Objective: Ischemia-reperfusion (I/R) leads to reactive oxygen species formation and cell death in kidney tissue with injury and organ transplantation. Simvastatin (SIM) is an antioxidant, anti-inflammatory, and anticoagulant agent. Alterations in I/R-induced acute kidney injury model with SIM treatment were analyzed.
Study Design: Wistar rats (n=28) were grouped into Sham, Ischemia, I/R, and I/R+SIM treated. Left rat kidney renal vessels were clamped for 60 minutes for ischemia, and the I/R group had 6 hours of reperfusion. 10 mg/kg SIM was given orally for 28 days. MDA, GSH, and MPO were analyzed. Kidney tissues were paraffin embedded, and primary antibodies TNF-α and caspase-3 were applied for immunohistochemistry.
Results: In the I/R group, intense inflammatory cell infiltration around the vessels and necrosis in the glomerular structures were observed. In the treated group, proximal and distal tubular cells were found to be close to normal. Immunoexpression of caspase-3 in the ischemia group was positive in degenerative glomeruli. In the treated group, TNF-α expression was negative in the glomerular structures. MDA and MPO levels were significantly increased in ischemia and I/R.
Conclusion: We suggest that SIM treatment improved kidney tissue structure and function in a model of I/R injury.
Keywords: caspase-3; immunohistochemistry; ischemia/reperfusion; kidney; MPO; simvastatin
Objective: The prognostic indictors of age-related poor outcomes in patients with acute myeloid leukemia (AML) are still controversial. The aim of this work was to provide comprehensive insights into the effect of different hemocytes and to investigate the association between age and clinical features in adult patients with AML.
Study Design: A retrospective study was performed to determine the role of age in the therapeutic outcomes of AML. A total of 166 newly diagnosed adult patients’ data from January 2015 to November 2019 in Zhongshan Hospital of Xiamen University were collected and analyzed.
Results: Older patients presented a poorer prognosis (p=0.001) with shorter overall survival, which is served as age-related outcomes. Binary logistic regression demonstrated that cytogenetic risk (OR=4.508, 95% CI 2.733–7.435), leukocyte (OR=7.410, 95% CI 1.139–5.910), and bone marrow blast cells (OR=3.261, 95% CI 1.075–5.615) were independent indictors for age-related prognosis. In addition, Kaplan-Meier curve also revealed that the above factors were associated with overall survival (all p values <0.001).
Conclusion: Cytogenetic risk, leukocyte, and bone marrow blast cells are dominant factors which account for the age-related poor outcomes and shorter overall survival in AML.
Keywords: acute myeloid leukemia, adult, cytogenetic risk, hemocyte, leukemia, overall survival
BACKGROUND: Sequential Epstein-Barr virus (EBV)–positive B cell lymphoma to the initial diagnosis of angioimmunoblastic T cell lymphoma (AITL) is very rare, the exact mechanism and standard therapy of which is still being explored. CASE: A 50-year-old man was admitted to our hospital in January 2014 with a three-week history of enlargement of multiple lymph nodes. His initial pathological evaluation indicated AILT. The reactivation of EBV was observed during the immunosuppression therapy for AITL, accompanied by onset of subcutaneous nodules proven to be EBV-positive diffuse large B cell lymphoma (DLBCL) based on the pathological findings of rebiopsy. The patient was successfully treated with chidamide, a histone deacetylase (HDAC) inhibitor, and rituximab.
Conclusion: The sufficient surveillance for serum EBV and repeat biopsy is necessary for patients with AITL, and this treatment modality may become an active option.
Keywords: angioimmunoblastic T cell lymphoma, Epstein-Barr virus, HDAC inhibitor, non-Hodgkin lymphoma, peripheral T cell lymphoma
Objective: To probe into the influence of miR-21 on the proliferation as well as apoptosis of oral squamous cell carcinoma (OSCC) and its causative role.
Study Design: We adopted microarray for detecting the differentially expressed genes in OSCC tumor tis-sues and paracancerous tissues. We assessed the link of miR-21 expression with tumor size, lymph node metastasis, and tumor differentiation. We employed CCK-8 and EdU assay for detecting the impact of miR-21 inhibitor and miR-21 mimic on Cal-27 cell proliferation, as well as TUNEL and AnnexinV-FITC/PI double staining for detecting miR-21 expression on cell apoptosis. We forecasted the possible target of miR-21 via TargetScan, as well as detected the interaction of miR-21 with PTEN via luciferase reporter experiment. The function of miR-21 expression in PTEN signaling pathway was monitored via western blot. We constructed PTEN overexpression plasmid and conducted rescue experiment to evaluate overexpressed PTEN on miR-21–induced proliferation.
Results: Microarray and RT-qPCR indicated that miR-21 expression increased demonstrably in OSCC. Subsequently, statistical analysis showed that miR-21 expression was plainly correlated with tumor size, lymph node metastasis, tumor differentiation, and smoking history. CCK-8 and EdU method exhibited that miR-21 mimics manifestly promoted Cal-27 cell proliferation, while miR-21 inhibitor blatantly inhibited Cal-27 cell proliferation. TUNEL and V-FITC/PI double staining assay showed that miR-21 inhibitor conspicuously promoted Cal-27 cell apoptosis. CCK-8 and EdU assay exhibited that overexpressed PTEN abolished the pro-proliferation influence of miR-21 mimic. TUNEL and V-FITC/PI experiments pointed out that knocking down PTEN abrogated the pro-apoptosis impact of miR-21 inhibitor.
Conclusion: miR-21 contributes to OSCC cell proliferation via targeting PTEN and inhibits its apoptosis.
Keywords: Akt/PKB signaling pathway; apoptosis; biomarkers, tumor; carcinoma, squamous cell; cell line, tumor; cell proliferation; microRNAs; miR-21; miRNA-21; mouth neoplasms; oral cancer; oral squamous cell carcinoma; proliferation; real time PCR
Objective: To analyze the sonographic features of different histopathological subtypes of borderline ovarian tumors (BOTs) confirmed by pathology, and to study the ultrasound performances of various types in borderline ovarian tumors.
Study Design: Retrospective analysis was performed on the pathological results and ultrasound projection findings of 129 patients diagnosed as BOTs by ultrasound department of our hospital from January 2012 to November 2019. All patients were confirmed by surgical pathology and scanned consecutively by the investigators using transabdominal or transvaginal ultrasound examination.
Results: Serous borderline tumors (SBOTs) were observed, and the prevalence rate (53%) was significantly higher than that of other subtypes, and the probability of bilateral lesions was higher (40%). The sonogram often showed ultrasound features of papillary neoplasm in the lesion and good internal echo (p<0.05). Mucinous borderline ovarian tumors (MBOTs) were mostly unilateral lesions (86%). The prevalence was second only to SBOTs. Histomorphological examinations were divided into gastrointestinal-type and endocervical-type. Among them, the gastrointestinal type of MBOTs were mostly unilateral, and their incidence was higher than that of endocervical-type of MBOTs. Compared with other pathological subtypes, the gastrointestinal type is more likely to show the sonographic characteristics of huge space occupying in the pelvic and abdominal cavity (mean diameter >10 cm), polycystic, multiple septums, and poor internal echo (p<0.05). The ultrasonographic features of the endocervical-type of MBOTs were similar to those of SBOTs. Compared with gastrointestinal type, the sonographic images showed smaller lesion diameter, less septal or cyst, and more papillary excrescences in the tumor (p<0.05). The borderline clear cell tumor is the intermediate transition between the clear cell adenofibroma and the clear cell carcinoma. The clinical manifestations are diverse and lack specificity. The histology of sonography was mainly solid, and the multiple microcapsules were honeycomb-like. It can also be shown as cystic. Among the 169 patients with BOTs, 20 cases of SBOTs, 17 cases of MBOTs, and 10 cases of other rare subtypes were complicated with other diseases or multiple subtypes. This study did not find significant ultrasonic characteristics were used for distinguish them from other subtypes.
Conclusion: BOTs is a common disease in women during the reproductive period. It is characterized by the development of malignant tumors. Its clinical and pathological subtypes are complex and diverse. It leads many doctors to use the terms “large pelvic mass” and “solid ovarian mass” for diagnosis because of their lack of experience and understanding.
Keywords: adenocarcinoma, mucinous; adenocarcinoma, serous; borderline ovarian tumors; diagnostic imaging; ovarian neoplasms; papillary neoplasms; prognosis; transvaginal ultrasound, ultrasonography
Objective: The association between telomerase reverse transcriptase (TERT) promoter mutation and outcome of melanoma is unclear and controversial. We aim to conduct a meta-analysis and investigate whether the TERT promoter mutation is a prognostic factor of melanoma.
Study Design: Appropriate studies were searched in 3 databases: PubMed, Web of Science, and Embase. Pooled hazard ratios (HRs) were counted through random effects model.
Results: Heterogeneity was moderate in overall survival (OS) (I2=43.7%, p=0.059) and low in disease-free survival (DFS) (I2=0.0%, p=0.587). Sensitivity analysis indicated that the removal of any of the study did not affect the final results. Evidence for publication bias was not found (Begg’s test, p=0.281; Egger’s test, p=0.078). The pooled OS HRs from combined effects analysis was determined (HR 1.07; 95% CI 0.83–1.39, p=0.585), together with the pooled HRs of DFS (HR 1.65; 95% CI 1.02–2.66, p=0.042). TERT promoter mutation predicted a good outcome in meta-static melanoma patients (HR 0.66; 95% CI 0.46–0.96, p=0.042). The pooled HRs of combined mutation in TERT promoter and BRAF (HR 6.27; 95% CI 2.7–14.58, p=0.000) predicted a bad outcome in melanoma patients.
Conclusion: TERT promoter mutation significantly predicted poor DFS outcome but, on the contrary, predicted a good outcome in metastatic melanoma patients. The combined TERT promoter and BRAF mutation was a significant independent factor of OS in melanoma patients.
Keywords: melanoma; meta-analysis; mutation; prognosis; promoter regions, genetic; skin neoplasms; telomerase; TERT promoter mutation; TERT protein, human
Objective: Ischemia-reperfusion (I/R) leads to reactive oxygen species formation and cell death in kidney tissue with injury and organ transplantation. Simvastatin (SIM) is an antioxidant, anti-inflammatory, and anticoagulant agent. Alterations in I/R-induced acute kidney injury model with SIM treatment were analyzed.
Study Design: Wistar rats (n=28) were grouped into Sham, Ischemia, I/R, and I/R+SIM treated. Left rat kidney renal vessels were clamped for 60 minutes for ischemia, and the I/R group had 6 hours of reperfusion. 10 mg/kg SIM was given orally for 28 days. MDA, GSH, and MPO were analyzed. Kidney tissues were paraffin embedded, and primary antibodies TNF-α and caspase-3 were applied for immunohistochemistry.
Results: In the I/R group, intense inflammatory cell infiltration around the vessels and necrosis in the glomerular structures were observed. In the treated group, proximal and distal tubular cells were found to be close to normal. Immunoexpression of caspase-3 in the ischemia group was positive in degenerative glomeruli. In the treated group, TNF-α expression was negative in the glomerular structures. MDA and MPO levels were significantly increased in ischemia and I/R.
Conclusion: We suggest that SIM treatment improved kidney tissue structure and function in a model of I/R injury.
Keywords: caspase-3; immunohistochemistry; ischemia/reperfusion; kidney; MPO; simvastatin
Objective: The prognostic indictors of age-related poor outcomes in patients with acute myeloid leukemia (AML) are still controversial. The aim of this work was to provide comprehensive insights into the effect of different hemocytes and to investigate the association between age and clinical features in adult patients with AML.
Study Design: A retrospective study was performed to determine the role of age in the therapeutic outcomes of AML. A total of 166 newly diagnosed adult patients’ data from January 2015 to November 2019 in Zhongshan Hospital of Xiamen University were collected and analyzed.
Results: Older patients presented a poorer prognosis (p=0.001) with shorter overall survival, which is served as age-related outcomes. Binary logistic regression demonstrated that cytogenetic risk (OR=4.508, 95% CI 2.733–7.435), leukocyte (OR=7.410, 95% CI 1.139–5.910), and bone marrow blast cells (OR=3.261, 95% CI 1.075–5.615) were independent indictors for age-related prognosis. In addition, Kaplan-Meier curve also revealed that the above factors were associated with overall survival (all p values <0.001).
Conclusion: Cytogenetic risk, leukocyte, and bone marrow blast cells are dominant factors which account for the age-related poor outcomes and shorter overall survival in AML.
Keywords: acute myeloid leukemia, adult, cytogenetic risk, hemocyte, leukemia, overall survival
Osteoblasts remotely supply lung tumors with cancer-promoting SiglecFhigh neu...Gul Muneer
Systemic cross-talk between lung tumors and bones
Bone marrow–derived myeloid cells can accumulate within tumors and foster
cancer outgrowth. Local immune-neoplastic interactions have been intensively
investigated, but the contribution of the systemic host environment to tumor growth
remains poorly understood. Here, we show in mice and cancer patients (n = 70) that
lung adenocarcinomas increase bone stromal activity in the absence of bone
metastasis. Animal studies reveal that the cancer-induced bone phenotype involves
bone-resident osteocalcin-expressing (Ocn+) osteoblastic cells. These cells promote
cancer by remotely supplying a distinct subset of tumor-infiltrating SiglecFhigh
neutrophils, which exhibit cancer-promoting properties. Experimentally reducing
Ocn+ cell numbers suppresses the neutrophil response and lung tumor outgrowth.
These observations posit osteoblasts as remote regulators of lung cancer and
identify SiglecFhigh neutrophils as myeloid cell effectors of the osteoblast-driven
protumoral response
Proteogenomic analysis of human colon cancer reveals new therapeutic opportun...Gul Muneer
We performed the first proteogenomic study on a prospectively collected colon cancer cohort. Comparative proteomic and phosphoproteomic analysis of paired tumor and normal adjacent tissues produced a catalog of colon cancer-associated proteins and phosphosites, including known and putative new biomarkers, drug targets, and cancer/testis antigens. Proteogenomic integration not only prioritized genomically inferred targets, such as copy-number drivers and mutation-derived neoantigens, but also yielded novel findings. Phosphoproteomics data associated Rb phosphorylation with increased proliferation and decreased apoptosis in colon cancer, which explains why this classical tumor suppressor is amplified in colon tumors and suggests a rationale for targeting Rb phosphorylation in colon cancer. Proteomics identified an association between decreased CD8 T cell infiltration and increased glycolysis in microsatellite instability-high (MSI-H) tumors, suggesting glycolysis as a potential target to overcome the resistance of MSI-H tumors to immune checkpoint blockade. Proteogenomics presents new avenues for biological discoveries and therapeutic development.
Anti-lymphangiogenic properties of mTOR inhibitors in head and neck squamous ...Enrique Moreno Gonzalez
Tumor dissemination to cervical lymph nodes via lymphatics represents the first step in the metastasis of head and neck squamous cell carcinoma (HNSCC) and is the most significant predictor of tumor recurrence decreasing survival by 50%. The lymphatic suppressing properties of mTOR inhibitors are not yet well understood.
Osteoblasts remotely supply lung tumors with cancer-promoting SiglecFhigh neu...Gul Muneer
Systemic cross-talk between lung tumors and bones
Bone marrow–derived myeloid cells can accumulate within tumors and foster
cancer outgrowth. Local immune-neoplastic interactions have been intensively
investigated, but the contribution of the systemic host environment to tumor growth
remains poorly understood. Here, we show in mice and cancer patients (n = 70) that
lung adenocarcinomas increase bone stromal activity in the absence of bone
metastasis. Animal studies reveal that the cancer-induced bone phenotype involves
bone-resident osteocalcin-expressing (Ocn+) osteoblastic cells. These cells promote
cancer by remotely supplying a distinct subset of tumor-infiltrating SiglecFhigh
neutrophils, which exhibit cancer-promoting properties. Experimentally reducing
Ocn+ cell numbers suppresses the neutrophil response and lung tumor outgrowth.
These observations posit osteoblasts as remote regulators of lung cancer and
identify SiglecFhigh neutrophils as myeloid cell effectors of the osteoblast-driven
protumoral response
Proteogenomic analysis of human colon cancer reveals new therapeutic opportun...Gul Muneer
We performed the first proteogenomic study on a prospectively collected colon cancer cohort. Comparative proteomic and phosphoproteomic analysis of paired tumor and normal adjacent tissues produced a catalog of colon cancer-associated proteins and phosphosites, including known and putative new biomarkers, drug targets, and cancer/testis antigens. Proteogenomic integration not only prioritized genomically inferred targets, such as copy-number drivers and mutation-derived neoantigens, but also yielded novel findings. Phosphoproteomics data associated Rb phosphorylation with increased proliferation and decreased apoptosis in colon cancer, which explains why this classical tumor suppressor is amplified in colon tumors and suggests a rationale for targeting Rb phosphorylation in colon cancer. Proteomics identified an association between decreased CD8 T cell infiltration and increased glycolysis in microsatellite instability-high (MSI-H) tumors, suggesting glycolysis as a potential target to overcome the resistance of MSI-H tumors to immune checkpoint blockade. Proteogenomics presents new avenues for biological discoveries and therapeutic development.
Anti-lymphangiogenic properties of mTOR inhibitors in head and neck squamous ...Enrique Moreno Gonzalez
Tumor dissemination to cervical lymph nodes via lymphatics represents the first step in the metastasis of head and neck squamous cell carcinoma (HNSCC) and is the most significant predictor of tumor recurrence decreasing survival by 50%. The lymphatic suppressing properties of mTOR inhibitors are not yet well understood.
Diagonsis of cancer through saliva.pptxZaidAhmad42
Human saliva is an ideal body fluid for developing non-invasive diagnostics. Saliva contains naturally-occurring nanoparticles with unique structural and biochemical characteristics.
Role of human papillomavirus and tumor suppressor genesishita1994
Oral cancer is synonymous to Squamous Cell Carcinoma (SCC) of oral mucosal origin that accounts for more than 90% of all malignant presentations at the aforementioned anatomical sites.
More than 300,000 new cases worldwide are being diagnosed with oral SCC (OSCC) annually.
Approximately, 30,000 (US) & 40,000(EUROPE).
Oral cancer is estimated by the WHO to be the 8th most common cancer worldwide.
In India & other Asian countries, oral & oropharyngeal carcinomas (OCs) comprise up to half of all malignancies, with this particularly high prevalence being attributed to the influence of carcinogens & region-specific epidemiological factors, especially tobacco & betel quid chewing.
Sex-Based Difference in Gene Alterations and Biomarkers in Anal Squamous Cell...semualkaira
anal squamous cell carcinoma (ASCC) is a relatively rare malignancy ac-counting for about 2-3% of all the gastrointestinal tumors. The standard of treatment for localized disease is chemoradiotherapy
Sex-Based Difference in Gene Alterations and Biomarkers in Anal Squamous Cell...semualkaira
anal squamous cell carcinoma (ASCC) is a relatively rare malignancy ac-counting for about 2-3% of all the gastrointestinal tumors. The standard of treatment for localized disease is chemoradiotherapy. Several studies reported a sex disparity
in ASCC prognosis showing a better survival for female compared
to men. Methods: we examined 1,380 patients with ASCC who received comprehensive genomic profiling as part of routine clinical
care and present key
Flavin-Containing Dimethylaniline Monooxygenase 5 Drives Malignancies in Hepa...semualkaira
Hepatic microsomes play an important role in drug metabolism,
but the potential biological functions of hepatic microsome-containing proteins in Hepatocellular Carcinoma (HCC) remain unclear. Here, we used HCC and corresponding adjacent Non-Tumor
(NT) tissues to isolate hepatic microsomes and then performed
RNA high-throughput sequencing
Flavin-Containing Dimethylaniline Monooxygenase 5 Drives Malignancies in Hepa...semualkaira
Hepatic microsomes play an important role in drug metabolism, but the potential biological functions of hepatic microsome-con- taining proteins in Hepatocellular Carcinoma (HCC) remain un- clear. Here, we used HCC and corresponding adjacent Non-Tumor (NT) tissues to isolate hepatic microsomes and then performed RNA high-throughput sequencing. After screening, flavin-con- taining dimethylaniline monooxygenase (FMO5) showed a significantly high expression level and was associated with poor prognosis in patients with HCC.
International Journal of Engineering and Science Invention (IJESI)
Similar to Silenced microRNA-135b-5p Inhibits Tongue Squamous Cell Carcinoma Proliferation and Apoptosis by Regulating ARID1A and the PI3K/AKT Axis (19)
Objective: To investigate the protective effect of lo- sartan, an angiotensin II type 1 receptor blocker with antioxidative effect on intestinal ischemia-reperfusion (I/R) injury in rats, against inflammation and apoptotic development.
Study Design: Forty male Wistar albino rats with a mean weight of 200–250 g each were divided into 4 groups: (1) Sham operation (laparotomy only, sham surgical preparation including isolation of the superior mesenteric artery [SMA] without occlusion), (2) Ischemia model with SMA closure for 2 hours, (3) I/R group (2 hours of ischemia followed by 3-hour reperfusion (SMA occlusion for 120 minutes followed by 240 minutes reperfusion), and (4) Losartan group (2 hours of ischemia, 40 mg/kg losartan was administered to the animals; losartan was dissolved in 1 mL distilled water and administered intraperitoneally after 2 hours of ischemia). Malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) levels were examined in jejunum tissue.
Results: Losartan treatment reduced the I/R-induced increase in MDA levels in the gut. Statistically, while SOD, CAT, and GSH activities decreased significantly in the I/R group, they increased in the I/R+Losartan group. Villus loss and increase in inflammation after ischemia persisted after reperfusion. Losartan treatment played a role in the reduction of inflammation and apoptosis and in the regulation of TNF-α and caspase-9 activity.
Conclusion: It has been thought that losartan in I/R may reduce mucosal damage and cell apoptosis in the direction of inflammation and may stabilize caspase-9 activity by inhibiting TNF-α stimulus.
Keywords: caspase-9, ischemia, ischemia/reperfusion, rat, reperfusion injury, TNF-α, tumor necrosis factor-alpha
Objective: In order to reduce complications accompanied with dental implant restoration, this study strives to prepare a novel sealant and lubricant that can be used in dental implant systems as well as to evaluate its characteristics.
Study Design: Chitosan (CS), β-glycerophosphate pentahydrate (β-GP), and nano silver (nAg) were used to prepare thermosensitive hydrogel. According to the different volume ratios of CS to β-GP, 3 experimental groups were established, namely 16/4, 13/7, and 10/10 groups. Their morphology, composition, and chemical properties were analyzed via SEM, EDS, and FTIR. In addition, the effect of the hydrogel on the stability of dental implant-abutment connection was investigated by removal torque test combined with dynamic cyclic loading experiment. The maximum fracture load was measured under different lubricating conditions by electronic universal testing machine. The cytotoxicity and in vitro antibacterial effect of the hydrogel were examined respectively by CCK-8 test and the spread plate method.
Results: The CS/β-GP/nAg thermosensitive hydro-gel was successfully prepared in this study, which was found to be a porous structure through SEM. The removal torque test and the dynamic cyclic loading experiment showed that the removal torque of the experimental group was greater than that of the control group. Furthermore, the single load-to-fracture test indicated that the 16/4 group had the greatest maximum bearing load. The in vitro cytotoxicity test using rat bone marrow stromal cells (rBMSCs) and human gingival fibroblast cells (hGFCs) showed no cytotoxicity in all 3 groups. The 3 experimental groups had obvious antibacterial effects against E. coli, S. aureus, and P. gingivalis.
Conclusion: A nontoxic antibacterial CS/β-GP/nAg thermosensitive hydrogel for lubricating purpose was successfully fabricated. When the volume ratio of CS to β-GP was 16/4, this thermosensitive hydrogel demonstrated better sealing and lubricating abilities and had a positive influence on the reliability of dental implant-abutment connection.
Keywords: abutment, dental implant, dental implant restoration, dental sealant, lubrication, thermosensitive hydrogel
Objective: To investigate the bond strength of resin-modified glass ionomer enhanced with bioactive glass (Activa BioActive-Base/Liner) to composite resin using different dental adhesive systems.
Study Design: In this study, Activa BioActive-Base/Liner (ABA/BL) was placed in cylindrical cavities formed in acrylic blocks. In blocks divided into 6 groups according to the adhesive system to be applied, two-step etch-and-rinse Gluma 2 Bond (Heraeus Kulzer, Germany), one-step self-etch Gluma Self Etch (Heraeus Kulzer), universal system Gluma Universal (Heraeus Kulzer), two-step self-etch Clearfil SE Protect (Kuraray, Japan), one-step self-etch Clearfil S3 Bond Plus (Kuraray), and universal system Clearfil S3 Bond Universal (Kuraray) adhesive systems were applied on ABA/BL. After composite resin (3M ESPE Filtek Ultimate) was applied to the prepared surfaces, the specimens were placed in a universal test device and shear bond strength test was determined. Fracture types were evaluated using a stereomicroscope and scanning electron microscope. Data were analyzed by Shapiro-Wilk, two-way ANOVA, Kruskal-Wallis, and Post-Hoc Multiple Comparisons tests.
Results: In terms of bond strength values, the highest bond value was seen in the two-step self-etch (Clearfil SE Protect) group, and the lowest bond strength value was seen in the universal system (Clearfil S3 Bond Universal) group. There was no statistically significant difference between the adhesive agent groups in terms of bond strength values (p>0.05).
Conclusion: It is thought that choosing the two-step self-etch technique as an adhesive system when resin-modified glass ionomer enhanced with bioactive glass (ABA/BL) is used as the pulp capping/base material will be more appropriate in terms of bond strength.
Keywords: adhesive systems, bioactive materials, bond strength, cariostatic agents, composite resins, dental materials, fluorides, glass ionomer, glass ionomer cements, materials testing, vital pulp therapy
Objective: To evaluate the results of the effect of nebivolol on tibial bone defect and graft application in new bone development in the rat.
Study Design: Thirty Wistar albino rats were divided into 3 groups. In the Control group, tibia bone defect was created without any treatment. In the Defect+ Graft group, allograft treatment was performed by forming a 6 mm tibial bone defect. In the Defect+Graft+ Nebivolol group, alloplastic bone graft was placed in the calvarial bone defect and then nebivolol (0.34 mg/mL solution/day) treatment was intraperitoneally applied for 28 days.
Results: Histopathological examination revealed inflammation in the defect area, congestion in the vessels, degeneration in collagen fibers, and an increase in osteoclast cells. There was an increase in inflammation and blood vessel structure in graft application, and osteoblastic activity matrix formation after reorganization nebivolol application in collagen fibers. Osteonectin expression was positive in the collagen fiber and matrix, starting in the Graft group, in osteoblasts, whereas in the Nebivolol group, osteoblasts increased in osteocytes and new bone formation.
Conclusion: Nebivolol is thought to have a positive effect on osteoinductive bone growth factors and contribute to the cell-matrix interaction, in addition to the supporting effect of the graft with its antioxidative effect.
Keywords: allograft; bone; bone regeneration; disease models, animal; nebivolol; orthopedic procedures; osteonectin; rats; tibia; tibial defect
Objective: To investigate the effects of nicorandil and tirofiban on no-reflow and postoperative outcome in patients with acute coronary syndrome (ACS) undergoing percutaneous coronary intervention.
Study Design: A total of 438 patients with ACS diagnosed by the second Hospital of Shanxi Medical University from January 2019 to December 2020 were divided into two groups: nicorandil group (n=223) and tirofiban group (n=215). The nicorandil group was injected with 2 mg nicorandil 2 mm before coronary occlusion before balloon dilation, and the tirofiban group received 10 μg/kg intravenous injection during operation. Measurement of thrombolysis grade (thrombolysis in myocardial infarction [TIMI]), corrected TIMI frame count, and major adverse vascular events were recorded 30 days after operation in patients with ACS.
Results: Both nicorandil and tirofiban could improve the TIMI grade, and TIMI grade 3 blood flow was obtained in 190 cases (85.2%) and 175 cases (81.4%), respectively. There was no significant difference in the incidence of major adverse cardiac events (14.3% vs. 13.5%, score 0.13).
Conclusion: Intracoronary use of nicorandil in patients with ACS can improve coronary perfusion, but the improvement of prognosis needs further study.
Keywords: coronary perfusion, myocardial infarction, nicorandil, no-reflow phenomenon, percutaneous coronary intervention, repercussion
Objective: To identify interstitial cells of Cajal (ICC) in the common bile duct of Kunming mice.
Study Design: Common bile ducts obtained from the Kunming mice were prepared for immunohistochemical investigations using the c-kit antibody. Immunoelectron microscopy was used to detect the expression of c-kit in the ICC of the common bile duct. Transmission electron microscopy showed ultrastructure of ICC in the murine bile duct. Reverse transcription–polymerase chain reaction (RT-PCR) and western blot were used to confirm the expression of mRNA specific for the c-kit gene and production of c-kit protein in the Kunming mice common bile duct.
Results: Immunohistochemistry revealed that ICC in the murine common bile duct are c-kit positive and the ICC are located in the tela submucosa and the tunica muscularis of the murine common bile duct and do not connect with each other. Immunoelectron microscopy confirmed the expression of Kit by ICC in the murine common bile duct. Transmission electron microscopy showed that ICC in the murine common bile duct have long processes, abundant mitochondria, plenty of smooth endoplasmic reticulum (sER), a lot of lysosomes, and dense bodies. The caveolae of ICC are distinctive. At the same time, RT-PCR indicated that the Kunming mice common bile duct expressed mRNA specific for the c-kit gene, and western blot analysis showed the evidence of production of c-kit protein in the Kunming mice common bile duct.
Conclusion: ICC are found in the Kunming mice common bile duct, which is likely to lead to the development of motility study of the common bile duct.
Keywords: common bile duct; electron microscopy; immuno-electron microscopy; interstitial cells of Cajal; intestines; smooth muscle; tyrosine kinase receptor (c-kit)
Objective: To study the effects of resveratrol in neuronal structures in traumatic brain injury (TBI).
Study Design: Thirty rats were categorized as (1) control group (n=10), saline solution administered i.p. for 14 days, (2) TBI group (n=10), trauma induced by weight-drop model on brain, and (3) TBI+Resveratrol group (n=10), 15 minutes after injury the rats were given resveratrol (10 μmoL/kg/i.p.) for 14 days. At the end of the experiment the cerebellum was excised for routine paraffin tissue protocol. Blood samples were tested for serum biochemical markers (MDA, SOD, CAT, and GSH-x).
Results: SOD, GPx, and CAT values were lowest in the TBI group. MDA and histological scores of dilations in vessels, inflammation, degeneration in neurons, apoptosis in microglia, ADAMTS8, and GFAP expressions were highest in the TBI group. Sections of the control group showed normal cerebellar histology. The trauma group showed degenerated ganglion layer, pyknotic and apoptotic Purkinje cell nuclei. Vascular thrombus was seen in the substantia alba and substantia grisea. In the Trauma+Resveratrol group, most pa- thologies observed in the TBI group were improved. In the control group, GFAP protein was expressed in granular cells, axons, dendrites, Purkinje cells, and microglia cells. In the trauma group, increased GFAP expression was observed in glial processes, neurons, and Purkinje cells. In the Trauma+Resveratrol group, GFAP was expressed in molecular layer and glial processes. In the control group, ADAMTS-4 activity was observed in granulosa layer, glial cells, and Purkinje cells. In the trauma group, ADAMTS-4 expression was positive in Purkinje cells and glial cells. In the Trauma+ Resveratrol group, ADAMTS-4 was expressed in Purkinje cells, granular cells, and glial cells.
Conclusion: GFAP and ADAMTS-4 proteins may be involved in regeneration of damaged astroglial cells and other glial cells, Purkinje cells, and synaptic extensions. We suggest that antioxidative drugs such as resveratrol may be alternative target agents in neurological disease.
Keywords: ADAMTS-4, brain, cerebellum, GFAP, rat, resveratrol, traumatic brain injury
Objective: To evaluate the antibacterial effects of 4 different cavity disinfectants on Streptococcus mutans, Lactobacillus acidophilus, and Enterococcus faecalis bacteria in different time periods.
Study Design: The antibacterial effects of Cavity Cleanser, Tubulicid Red Label, Chloraxid 2%, and Oxygenated Water cavity disinfectant solutions on E. faecalis (ATCC 29212), S. mutans (ATCC 25175), and L. acidophilus (RSKK 03037) bacterial strains were evaluated by disk diffusion method. In the study where vancomycin antibiogram disc constituted the positive control group, physiological saline solution was used as the negative control group. Standard, sterile, blank antibiogram discs of 5 mm in diameter, in which 15 μL of each material were added, were placed on agar plates at 2.5–3 cm intervals. The inhibition zone diameters formed around the discs that were left to incubate for 24–48 hours at 37°C were measured in millimeters. Statistical analysis of the data was performed using one-way analysis of variance, Kolmogorov-Smirnov, Levene, and Bonferroni tests.
Results: At the end of the study the solutions tested showed a statistically significant antibacterial effect on all bacterial strains used (p<0.05). Cavity Cleanser disinfectant containing 2% chlorhexidine showed the highest antibacterial effect on S. mutans and L. acidophilus, and benzalkonium-containing Tubulicid Red disinfectant on E. faecalis.
Conclusion: The antibacterial effect of all cavity disinfectants used in the study was found to be higher at the end of the 48th hour than at the end of the 24th hour, but there was no statistically significant difference (p>0.05).
Keywords: antibacterial agents; antibacterial effect; cavity disinfectants; chlorhexidine; contamination; dental caries; disinfection; disc diffusion; gram-negative bacteria; gram-positive bacteria
Objective: To investigate the changes in the retina due to deltamethrin toxicity and the process in cell inflammation and apoptosis.
Study Design: Sixteen Wistar albino rats were randomly divided into two groups as control (n=8) and deltamethrin (n=8) groups. Saline was given to the control group, and 0.5 mL of 5 mg/kg deltamethrin was given to the deltamethrin group for 14 days each. Blood was collected for biochemical analysis. Retinal tissue was processed for histological examination.
Results: Compared to the control group, MDA levels were high while GSH and CAT levels were low in the deltamethrin group. Histopathological analysis showed spaces between the pigment epithelium, irregularity in the delimiting membrane, degenerated ganglion, cone and bacillus cell, pyknotic nuclei, thinned inner limitation membrane, and thickened vascular wall. The control group showed FAS expression in the pigment layer limiting membranes, in the nuclei of many cone and bacillus cells, and ganglion cells in the control group sections. In the deltamethrin group, FAS expression was observed in the inner and outer limiting membranes of the pigment epithelium, cone and bacillus cells, and ganglion cell nuclei. In the control group, negative NOS expression in the pigment epithelium and outer limiting membranes, internal limitation membrane, and ganglion cells in the cone and bacillus cell nuclei were observed. In the deltamethrin group, NOS expression was positive in the pigment epithelium, cone and bacillus, and ganglion cell nuclei.
Conclusion: We suggest that deltamethrin toxicity induced apoptotic process due to increased inflammation in the retina and may cause visual impairment as a result of neural damage.
Keywords: deltamethrin, FAS, insecticides, NOS, nitric oxide synthase, retina
Objective: To investigate the immunohistochemical staining of hypoxia-inducible factor 1-alpha (HIF-1α) and Ki-67 expression in the placenta of pregnant women with placenta previa and placenta accreta.
Study Design: Thirty placentas (10 normotensive, 10 placenta previa, and 10 placenta accreta) were processed for routine histological tissue processing. The biochemical parameters of patients were recorded. Placentas were stained with hematoxylin-eosin and HIF-1α and Ki-67 immunostaining.
Results: Normal histology was observed in placentas of normotensive pregnant women. Placenta previa sections showed increased syncytial knots, intervillous hemorrhage, fibrin accumulation, and hyalinization. In placenta accreta sections, increased syncytial nodes, vascular dilation/congestion, fibrin accumulation, and hyalinization were observed. Normotensive placentas showed no HIF-1α expression. In placenta previa tissues, high HIF-1α expression was observed in vascular endothelial cells, villous stromal cells, and syncytial knots. High HIF-1α expression was recorded in villous stromal cells and cytotrophoblast cells in placenta accreta. In normotensive placental tissues, no Ki-67 expression was observed. In placenta previa sections, high Ki-67 expression was observed mostly in root villi stromal cells and some endothelial cells. High Ki-67 expression was observed mostly in villi stromal cells of placenta accreta.
Conclusion: It is thought that HIF-1α is an important regulatory gene in the development of villus in trophoblast invasion such as placenta accreta and previa, while Ki-67 will play a key role in the development of abnormal placenta with its stimulating effect on inflammatory cell development and angiogenesis in accreta and preeclampsia.
Objective: A spinal cord injury (SCI) is damage to the spinal cord either from trauma, loss of its normal blood supply, or compression from tumor or infection. In this study we focused on alterations in the bladder tissue with angiogenic and apoptotic aspects after spinal cord injury.
Study Design: Twenty Wistar Albino rats were categorized as control and SCI groups. At T7-T9 vertebras, a steel rod was dropped from 10 cm to create a spinal cord injury under anesthesia. Rats were decapitated and spinal tissue was processed to measure malondialdehyde (MDA), glutathione (GSH), and myeloperoxidase (MPO).
Results: MDA, MPO, epithelial degeneration, vascular dilation, inflammation, VEGF, and APAF-1 expressions in the SCI group were statistically higher than those in the control group. GSH content of the SCI group was statistically lower than that in the control group. In the hematoxylin-eosin–stained sections of the control group, normal histology was observed in bladder tissue. In the SCI group, degeneration epithelial cells, thinned epithelium, increased fibrosis, dilated and congested blood vessels, and hyperplastic endothelial cells were observed. In the control group, VEGF expression was slightly observed in some epithelial cells and vascular cells. In the SCI group, VEGF expression was increased in inflammatory and vascular endothelial cells. For APAF-1 expression, the control group showed no expression. In the SCI group, APAF-1 expression was positive in degenerated epithelial cells and connective tissue cells.
Conclusion: It is thought that the urination reflex was affected due to increased inflammation in the bladder tissue, leading to alterations in the regulation and function of the muscles.
Objective: To investigate the effect of sildenafil on reducing the impact of hepatic ischemia/reperfusion (HIR) injury established by Pringle maneuver on the heart of rats.
Study Design: Forty Wistar albino rats were divided into 4 groups: Sham (laparotomy only), Control (laparotomy following sildenafil application), IR (ischemia/reperfusion injured by HIR), and IR+SIL (injured by HIR following sildenafil application). Ischemia was developed by clamping the hepatoduodenal ligament for 30 minutes; then reperfusion was applied for 30 minutes. Sildenafil (single dose of 50 mg/kg) was administered by oral gavage for 15 minutes before ischemia. Blood samples of rats were collected from Sham and Control groups at 60 minutes and from IR and IR+SIL groups at 30 minutes after initiation of reperfusion for biochemical analysis. Meanwhile, heart tissues were sampled for biochemical analysis. Malondialdehyde (MDA) and total antioxidant capacity (TAC) in serum samples and TAC, total oxidative capacity (TOC), and oxidative stress index in heart tissues were examined biochemically.
Results: Serum MDA levels were elevated significantly in the IR and IR+SIL groups as compared to the sham group. Sildenafil treatment inhibited MDA increase considerably in the IR+SIL group as compared to the IR group. Serum TAC levels were elevated significantly in the sildenafil and control groups (compared with sham groups) and in the IR+SIL group (compared with the IR group). TAC levels detected in heart tissue increased significantly in the IR group as compared to the sham group; however, sildenafil treatment had no effect on this increase.
Conclusion: Heart tissue was affected by HIR. It was revealed that sildenafil treatment may prevent the oxidative stress via increasing serum TAC levels in both control and IR+SIL groups.
Objective: To examine the oropharynx of patients with ectodermal dysplasia showing maxillary retrusion and mandibular protrusion with a short and concave facial structure using cone-beam computed tomography method. Ectodermal dysplasia refers to the congenital disorder defined by the abnormal development of the structure originating from the ectoderm.
Study Design: In order to examine the oropharynx airway, measurements and statistical evaluations were made in 3 levels in sagittal and transversal directions on three-dimensional cone beam computed tomography images obtained from 14 individuals divided into 2 groups as Ectodermal Dysplasia group (n=7) and Control group (n=7).
Results: As a result of statistical analysis, no statistically significant difference was found between the groups at any level or direction in metric measurements performed on all 3 planes taken at the sagittal and transversal levels (p>0.05).
Conclusion: Our findings on ectodermal dysplasia are similar to Class III malpositions that show similarity with ectodermal dysplasia.
Objective: Diabetic nephropathy is one of the most serious complications of diabetes mellitus. It develops in approximately one-third of diabetic patients, years after the onset of metabolic abnormalities.
Study Design: The biopsy specimens were evaluated with the focus on light microscopy. The aim of our study was to reveal differences in the details and the frequency of occurrence of individual histomorphological changes in diabetic nephropathy and other glomerulonephritides.
Results: Diabetic nephropathy accounted for 14 out of 82 analyzed biopsies. Isolated thickening of the glomerular basement membrane was not present in any case, but along with some degree of mesangial expansion, hypercellularity or glomerulosclerosis was seen in 12 out of 14 findings of diabetic nephropathy. In other glomerular diseases, mesangial changes, but without glomerular basement membrane thickening, were the most frequent findings. In addition to glomerular lesions, some of the tubular, interstitial, and vascular changes were seen in 13 out of 14 patients with diabetic nephropathy. In other glomerulonephritides the combination of all these changes was a rare finding.
Conclusion: There are cases where immunofluorescence and electron microscopy cannot be performed or their results are not helpful. In such cases we must rely on light microscopic histomorphological changes.
Flu Vaccine Alert in Bangalore Karnatakaaddon Scans
As flu season approaches, health officials in Bangalore, Karnataka, are urging residents to get their flu vaccinations. The seasonal flu, while common, can lead to severe health complications, particularly for vulnerable populations such as young children, the elderly, and those with underlying health conditions.
Dr. Vidisha Kumari, a leading epidemiologist in Bangalore, emphasizes the importance of getting vaccinated. "The flu vaccine is our best defense against the influenza virus. It not only protects individuals but also helps prevent the spread of the virus in our communities," he says.
This year, the flu season is expected to coincide with a potential increase in other respiratory illnesses. The Karnataka Health Department has launched an awareness campaign highlighting the significance of flu vaccinations. They have set up multiple vaccination centers across Bangalore, making it convenient for residents to receive their shots.
To encourage widespread vaccination, the government is also collaborating with local schools, workplaces, and community centers to facilitate vaccination drives. Special attention is being given to ensuring that the vaccine is accessible to all, including marginalized communities who may have limited access to healthcare.
Residents are reminded that the flu vaccine is safe and effective. Common side effects are mild and may include soreness at the injection site, mild fever, or muscle aches. These side effects are generally short-lived and far less severe than the flu itself.
Healthcare providers are also stressing the importance of continuing COVID-19 precautions. Wearing masks, practicing good hand hygiene, and maintaining social distancing are still crucial, especially in crowded places.
Protect yourself and your loved ones by getting vaccinated. Together, we can help keep Bangalore healthy and safe this flu season. For more information on vaccination centers and schedules, residents can visit the Karnataka Health Department’s official website or follow their social media pages.
Stay informed, stay safe, and get your flu shot today!
Title: Sense of Taste
Presenter: Dr. Faiza, Assistant Professor of Physiology
Qualifications:
MBBS (Best Graduate, AIMC Lahore)
FCPS Physiology
ICMT, CHPE, DHPE (STMU)
MPH (GC University, Faisalabad)
MBA (Virtual University of Pakistan)
Learning Objectives:
Describe the structure and function of taste buds.
Describe the relationship between the taste threshold and taste index of common substances.
Explain the chemical basis and signal transduction of taste perception for each type of primary taste sensation.
Recognize different abnormalities of taste perception and their causes.
Key Topics:
Significance of Taste Sensation:
Differentiation between pleasant and harmful food
Influence on behavior
Selection of food based on metabolic needs
Receptors of Taste:
Taste buds on the tongue
Influence of sense of smell, texture of food, and pain stimulation (e.g., by pepper)
Primary and Secondary Taste Sensations:
Primary taste sensations: Sweet, Sour, Salty, Bitter, Umami
Chemical basis and signal transduction mechanisms for each taste
Taste Threshold and Index:
Taste threshold values for Sweet (sucrose), Salty (NaCl), Sour (HCl), and Bitter (Quinine)
Taste index relationship: Inversely proportional to taste threshold
Taste Blindness:
Inability to taste certain substances, particularly thiourea compounds
Example: Phenylthiocarbamide
Structure and Function of Taste Buds:
Composition: Epithelial cells, Sustentacular/Supporting cells, Taste cells, Basal cells
Features: Taste pores, Taste hairs/microvilli, and Taste nerve fibers
Location of Taste Buds:
Found in papillae of the tongue (Fungiform, Circumvallate, Foliate)
Also present on the palate, tonsillar pillars, epiglottis, and proximal esophagus
Mechanism of Taste Stimulation:
Interaction of taste substances with receptors on microvilli
Signal transduction pathways for Umami, Sweet, Bitter, Sour, and Salty tastes
Taste Sensitivity and Adaptation:
Decrease in sensitivity with age
Rapid adaptation of taste sensation
Role of Saliva in Taste:
Dissolution of tastants to reach receptors
Washing away the stimulus
Taste Preferences and Aversions:
Mechanisms behind taste preference and aversion
Influence of receptors and neural pathways
Impact of Sensory Nerve Damage:
Degeneration of taste buds if the sensory nerve fiber is cut
Abnormalities of Taste Detection:
Conditions: Ageusia, Hypogeusia, Dysgeusia (parageusia)
Causes: Nerve damage, neurological disorders, infections, poor oral hygiene, adverse drug effects, deficiencies, aging, tobacco use, altered neurotransmitter levels
Neurotransmitters and Taste Threshold:
Effects of serotonin (5-HT) and norepinephrine (NE) on taste sensitivity
Supertasters:
25% of the population with heightened sensitivity to taste, especially bitterness
Increased number of fungiform papillae
Couples presenting to the infertility clinic- Do they really have infertility...Sujoy Dasgupta
Dr Sujoy Dasgupta presented the study on "Couples presenting to the infertility clinic- Do they really have infertility? – The unexplored stories of non-consummation" in the 13th Congress of the Asia Pacific Initiative on Reproduction (ASPIRE 2024) at Manila on 24 May, 2024.
Ozempic: Preoperative Management of Patients on GLP-1 Receptor Agonists Saeid Safari
Preoperative Management of Patients on GLP-1 Receptor Agonists like Ozempic and Semiglutide
ASA GUIDELINE
NYSORA Guideline
2 Case Reports of Gastric Ultrasound
Prix Galien International 2024 Forum ProgramLevi Shapiro
June 20, 2024, Prix Galien International and Jerusalem Ethics Forum in ROME. Detailed agenda including panels:
- ADVANCES IN CARDIOLOGY: A NEW PARADIGM IS COMING
- WOMEN’S HEALTH: FERTILITY PRESERVATION
- WHAT’S NEW IN THE TREATMENT OF INFECTIOUS,
ONCOLOGICAL AND INFLAMMATORY SKIN DISEASES?
- ARTIFICIAL INTELLIGENCE AND ETHICS
- GENE THERAPY
- BEYOND BORDERS: GLOBAL INITIATIVES FOR DEMOCRATIZING LIFE SCIENCE TECHNOLOGIES AND PROMOTING ACCESS TO HEALTHCARE
- ETHICAL CHALLENGES IN LIFE SCIENCES
- Prix Galien International Awards Ceremony
Acute scrotum is a general term referring to an emergency condition affecting the contents or the wall of the scrotum.
There are a number of conditions that present acutely, predominantly with pain and/or swelling
A careful and detailed history and examination, and in some cases, investigations allow differentiation between these diagnoses. A prompt diagnosis is essential as the patient may require urgent surgical intervention
Testicular torsion refers to twisting of the spermatic cord, causing ischaemia of the testicle.
Testicular torsion results from inadequate fixation of the testis to the tunica vaginalis producing ischemia from reduced arterial inflow and venous outflow obstruction.
The prevalence of testicular torsion in adult patients hospitalized with acute scrotal pain is approximately 25 to 50 percent
Recomendações da OMS sobre cuidados maternos e neonatais para uma experiência pós-natal positiva.
Em consonância com os ODS – Objetivos do Desenvolvimento Sustentável e a Estratégia Global para a Saúde das Mulheres, Crianças e Adolescentes, e aplicando uma abordagem baseada nos direitos humanos, os esforços de cuidados pós-natais devem expandir-se para além da cobertura e da simples sobrevivência, de modo a incluir cuidados de qualidade.
Estas diretrizes visam melhorar a qualidade dos cuidados pós-natais essenciais e de rotina prestados às mulheres e aos recém-nascidos, com o objetivo final de melhorar a saúde e o bem-estar materno e neonatal.
Uma “experiência pós-natal positiva” é um resultado importante para todas as mulheres que dão à luz e para os seus recém-nascidos, estabelecendo as bases para a melhoria da saúde e do bem-estar a curto e longo prazo. Uma experiência pós-natal positiva é definida como aquela em que as mulheres, pessoas que gestam, os recém-nascidos, os casais, os pais, os cuidadores e as famílias recebem informação consistente, garantia e apoio de profissionais de saúde motivados; e onde um sistema de saúde flexível e com recursos reconheça as necessidades das mulheres e dos bebês e respeite o seu contexto cultural.
Estas diretrizes consolidadas apresentam algumas recomendações novas e já bem fundamentadas sobre cuidados pós-natais de rotina para mulheres e neonatos que recebem cuidados no pós-parto em unidades de saúde ou na comunidade, independentemente dos recursos disponíveis.
É fornecido um conjunto abrangente de recomendações para cuidados durante o período puerperal, com ênfase nos cuidados essenciais que todas as mulheres e recém-nascidos devem receber, e com a devida atenção à qualidade dos cuidados; isto é, a entrega e a experiência do cuidado recebido. Estas diretrizes atualizam e ampliam as recomendações da OMS de 2014 sobre cuidados pós-natais da mãe e do recém-nascido e complementam as atuais diretrizes da OMS sobre a gestão de complicações pós-natais.
O estabelecimento da amamentação e o manejo das principais intercorrências é contemplada.
Recomendamos muito.
Vamos discutir essas recomendações no nosso curso de pós-graduação em Aleitamento no Instituto Ciclos.
Esta publicação só está disponível em inglês até o momento.
Prof. Marcus Renato de Carvalho
www.agostodourado.com
Pulmonary Thromboembolism - etilogy, types, medical- Surgical and nursing man...VarunMahajani
Disruption of blood supply to lung alveoli due to blockage of one or more pulmonary blood vessels is called as Pulmonary thromboembolism. In this presentation we will discuss its causes, types and its management in depth.
Report Back from SGO 2024: What’s the Latest in Cervical Cancer?bkling
Are you curious about what’s new in cervical cancer research or unsure what the findings mean? Join Dr. Emily Ko, a gynecologic oncologist at Penn Medicine, to learn about the latest updates from the Society of Gynecologic Oncology (SGO) 2024 Annual Meeting on Women’s Cancer. Dr. Ko will discuss what the research presented at the conference means for you and answer your questions about the new developments.
Knee anatomy and clinical tests 2024.pdfvimalpl1234
This includes all relevant anatomy and clinical tests compiled from standard textbooks, Campbell,netter etc..It is comprehensive and best suited for orthopaedicians and orthopaedic residents.
ARTIFICIAL INTELLIGENCE IN HEALTHCARE.pdfAnujkumaranit
Artificial intelligence (AI) refers to the simulation of human intelligence processes by machines, especially computer systems. It encompasses tasks such as learning, reasoning, problem-solving, perception, and language understanding. AI technologies are revolutionizing various fields, from healthcare to finance, by enabling machines to perform tasks that typically require human intelligence.
2. ating ARID1A was demonstrated by functional rescue
experiment.
RESULTS: miR-135b-5p was upregulated in TSCC
tissues and cells, while ARID1A was suppressed (p<
0.05). Silenced miR-135b-5p discouraged TSCC cell
proliferation, improved apoptosis, induced cell cycle
arrest, and increased ARID1A expression while inac-
tivating the PI3K/AKT axis (p<0.05). Furthermore,
knockdown of ARID1A reversed the impacts on TSCC
cell proliferation and apoptosis exerted by silencing
miR-135b-5p.
CONCLUSION: This research supported that silenced
miR-135b-5p impeded TSCC proliferation and apoptosis
by promoting ARID1A and inactivating the PI3K/AKT
axis, which may provide some indications for TSCC
alleviation. (Anal Quant Cytopathol Histpathol 2021;
43:151–160)
Keywords: apoptosis; ARID1A; ARID1A protein,
human; carcinoma, squamous cell; cell line, tu-
mor; cell proliferation; drug resistance, neoplasm;
microRNA-135b-5p; microRNAs; PI3K/AKT path-
way; neoplasm metastasis; neoplastic stem cells;
proliferation; protein binding; tongue; tongue squa-
mous cell carcinoma.
Tongue squamous cell carcinoma (TSCC) refers to
a prevailing type of oral SCC which is character-
ized by frequent metastasis of lymph node.1 Alter-
ations of tumor microenvironment, immune cells,
and checkpoints destruct immune order, making
tumors more evasive to medical observation.2 As
an oral cancer with high aggressiveness, TSCC
takes responsibility for about 41% of oral cancers.3
A mounting range of population at or under the
age of 40, young adults and women are especial-
ly at risk of being susceptible to TSCC globally.4
The pathogenesis of TSCC is tightly associated
with tobacco abuse, excessive alcohol, and infec-
tion with human papillomavirus.5 Immune thera-
pies such as durvalumab, pembrolizumab, atezo
lizumab, and nivolumab currently appear to be
promising and efficient treatments for TSCC.2
Although there are plenty of technologies, TSCC
remains a fatal malignancy in humans.3 In this
circumstance, we undertook a microRNA (miR)-
based approach to understand the underlying
mechanism in TSCC development in order to de-
velop novel intervention strategies.
To learn more about the biomarkers during
TSCC progression would be conducive to early di-
agnosis, proper treatment, and constructive prog-
nosis.6 miRs are suggested to be reliable biomark-
ers in cancer mitigation with their ability of varying
from oncogenes to tumor suppressors in various
tumor progression.7 As major modulators in piv-
otal biological activities, miRs also significantly
participate in cutaneous SCC (CSCC), making them
indispensable biomarkers in other SCC malignan-
cies.1,8 Although there are multiple discussions
about the protective or detrimental impacts of
different miRs on TSCC, the exact role of miR-
135b-5p in this type of cancer remains to be elu
cidated. In fact, in esophageal SCC (ESCC), miR-
135b-5p is already ranked as a classifier whose col-
laboration with clinicopathological prognostic tar-
gets could motivate the accuracy to prognosis pre-
diction.9 miR-135b is highly expressed in several
CSCC cell lines.10 Furthermore, evidence has sug-
gested that miR-135b-5p overexpression predicts
a poor survival time in ESCC patients.11 As an
oncogene in a large number of other severe can-
cers, including pancreatic cancer, gastric cancer,
and breast cancer, miR-135b-5p gives a push to
tumor metastasis and growth by suppressing its
target genes.12-14 It is also worth mentioning that
miR-135b upregulates a downstream factor to en-
courage colony formation, angiogenesis, and sur-
vival of head and neck SCC (HNSCC).15 Accord-
ing to the evidence mentioned above, we make a
hypothesis that miR-135b-5p may play an import-
ant role in TSCC development through interacting
with other genes.
Materials and Methods
Ethics Statement
This study was approved and supervised by
the ethics committee of First Affiliated Hospital,
Shihezi University School of Medicine. All of the
subjects signed the informed consent. The proto-
col was also approved by the Institutional Animal
Care and Use Committee of First Affiliated Hospi-
tal, Shihezi University School of Medicine.
Clinical Samples
A total of 52 subjects with TSCC who were di-
agnosed and received tumor resection at First
Affili
ated Hospital, Shihezi University School of
Medicine, from February 2016 to February 2019
were recruited in this study, with their TSCC
tumor tissues and adjacent normal tissues har-
vested. Inclusion criteria were that no patient
should have received radiotherapy, chemotherapy,
or any other targeted therapies before surgery,
152 Analytical and Quantitative Cytopathology and Histopathology®
Guo et al
3. and all the tissues should have been removed into
−80°C liquid nitrogen as soon as the surgery was
performed.
Cell Cultivation and Transient Transfection
Following a previous research, normal human
oral keratinocytes and TSCC cell lines, Cal27, SCC4,
and UM1 (American Type Culture Collection
[ATCC], Rockville, Maryland) were all cultured in
Dulbecco’s Modified Eagle medium (Gibco, Grand
Island, New York), consisting of fetal bovine se-
rum (HyClone, Logan, Utah), penicillin, and strep-
tomycin.16
Inhibitor negative control (NC), miR-135b-5p
inhibitor, small interfere (si)-NC or si-AT-rich
interactive domain‑containing protein 1A gene
(ARID1A) (GenePharma Co., Ltd. Shanghai, China)
were respectively transfected into Cal27 cells by
a Lipofectamine 2000 (Invitrogen, Thermo Fisher
Scientific Inc., Waltham, Massachusetts, USA) at
50 nM at 5×106 cells. After 48 hours of transfec-
tion, the remaining operations were conducted.17
Cell Counting Kit-8 (CCK-8) Method
Cells (2.5×104 cells/mL) were seeded into 96-well
plates and cultivated in a 37°C incubator with 5%
CO2 at 0 hour, 24 hours, 48 hours, and 72 hours,
respectively. Cells were incubated with 10 μL
CCK-8 (Dojindo Laboratories, Kumamoto, Japan)
in each well for 2 hours. After that, the optical den-
sity at 450 nm was observed.
Flow Cytometry
Cells were harvested and then washed twice with
phosphate-buffered saline (PBS). The cell cycle
assessment went as follows: cells were treated in
4°C 70% ethyl alcohol for 2 hours, washed by PBS,
cultured with dye buffer, interacted with propidi-
um iodide, and supplemented with RNase A with
out exposure to the light for 30 minutes. Then the
cells were evaluated via fluorescence-activated cell
sorting (FACS) by flow cytometry (AceaBio, San
Diego, California, USA), and cell apoptosis was
analyzed in accordance with the instructions of
Annexin V-FITC apoptosis detection kit (Beyotime
Biotechnology Co. Ltd., Shanghai, China). FACS
was also applied in cell apoptosis analysis.
Reverse Transcription–Quantitative Polymerase
Chain Reaction (RT-qPCR)
Trizol (Takara Biotechnology Ltd., Dalian, China)
was utilized to extract the total RNA in tissues
and cells to determine the concentration and purity
of RNA. RNA was reversely transcribed to cDNA
using PrimeScript RT kits (Takara Biotechnolo-
gy). Fluorescence qPCR was conducted with the
ABI PRISM 7300 system (Applied Biosystems, Inc.,
Carlsbad, California, USA) and the SYBR Premix
Ex Taq kits (Takara Biotechnology). PCR condi-
tions were as follows: pre-denaturating at 94°C
for 4 minutes (30 cycles of denaturating at 94°C
for 30 seconds, annealing at 59°C, and extension
at 72°C for 1 minute), and then extension at 72°C
for 5 minutes. And 2-ΔΔCt method was applied for
expression calculation of genes with glyceralde-
hyde 3-phosphate dehydrogenase (GAPDH) or U6
as the internal reference.18 The primers (Table I)
used were designed at Sangon Biotech Co., Ltd.
(Shanghai, China).
Western Blot Analysis
Cells were collected, and their proteins were
extracted using radio-immunoprecipitation assay.
Then, protein concentration was detected by bi-
cinchoninic acid protein assay kits (Beyotime).
Then, the total proteins were transferred onto the
polyvinylidene fluoride membranes (Amersham
Pharmacia Biotech, Piscataway, New Jersey, USA)
after being separated by 8% sodium dodecyl sul-
fate polyacrylamide gel electrophoresis. Subse-
quently, the membranes were sealed by 5% skim
milk powder for 2 hours and then incubated with
the primary antibodies: ARID1A (1:2000, ab264171),
GAPDH (1:1000, ab8245) (both from Abcam Inc.,
Cambridge, Massachusetts, USA), protein kinase
B (AKT) (1:1000, AF6261), p-AKT (1:1000, AF016)
(both from Affinity Biosciences, Cincinnati, Ohio,
USA), phosphatidyl inositol 3-kinase (PI3K) (1:1000,
CY5355), and p-PI3K (1:1000, CY6427) (both from
Volume 43, Number 4/August 2021 153
miR-135b-5p in TSCC Progression
Table I Primers Sequence of RT-qPCR
Gene Sequence (5’-3’)
miR-135b-5p F: 5’-GGTATGGCTTTTCATTCCT-3’
R: 5’-GCGAGCACAGAATTAATACGAC-3’
U6 F: 5’-GCTTCGGCAGCACATATACTAAAAT-3’
R: 5’-CGCTTCACGAATTTGCGTGTCAT-3’
ARID1A F: 5’-ACTCCATGGGGAGCTAGGT-3’
R: 5’-CACCCATGGGGTTTATGCCT-3’
GAPDH F: 5’-ACCCAGAAGACTGTGGATGG-3’
R: 5’-TCTAGACGGCAGGTCAGGTC-3’
ARID1A = AT-rich interactive domain‑containing protein 1A gene,
F = forward, GAPDH = glyceraldehyde 3-phosphate dehydrogenase,
miR = microRNA, R = reverse, RT-qPCR = reverse transcription–
quantitative polymerase chain reaction.
4. Abways, Shanghai, China) at 4°C overnight. Next,
the membranes were washed with tris-buffered
saline–tween buffer 3 times and cultured with
horse-radish peroxidase labeled goat anti-rabbit
immunoglobulin G antibody (1:2000; ab6721) (Ab
cam) for 1 hour. In addition, proteins were detect-
ed by strengthened electrochemiluminescence.
Dual Luciferase Reporter Gene Assay
Bioinformatics software was performed to deter-
mine the binding site between miR-135b-5p and
ARID1A 3’ untranslated region (3’UTR), on which
ARID1A wild-type (WT) and ARID1A mutant-type
(MUT) were constructed. 293T cells (ATCC) at
logarithmic growth phase were seeded into 96-
well plates. When the cell density approached 70%,
ARID1A-WT or ARID1A-MUT was mixed with
mimic NC and miR-135b-5p mimic (GenePharma)
and then co-transfected into Cal27 cells. After 48
hours transfection, cells were collected and lysed.
Luciferase activity was observed using luciferase
assay kits (BioVision, San Francisco, California,
USA) and a GloMax 20/20 Luminometer fluo-
rescence detector (Promega, Madison, Wisconsin,
USA). The experiment was repeated 3 times.
Statistical Analysis
SPSS 21.0 (IBM Corp., Released 2012, IBM SPSS
Statistics for Windows, Version 21.0. Armonk, New
York, USA) was employed to analyze data. The
data were shown in mean±standard deviation.
The t test was used for analyzing comparisons be-
tween two groups, one-way analysis of variance
(ANOVA) for comparing different groups, and
Tukey’s multiple comparisons test for pairwise
comparisons after ANOVA. The p value was at-
tained using a two-tailed test and p<0.05 referred
to significant difference.
Results
miR-135b-5p Is Highly Expressed in TSCC Tumor
Tissues and Cells
As the results of RT-qPCR represented, miR-135b-
5p expression was overtly higher in tissues of the
TSCC patients than in those of the adjacent nor-
mal tissues (p<0.05) (Figure 1A). Besides, com-
pared to that of the normal human oral keratino-
cytes, miR-135b-5p was strongly expressed in the
TSCC cell lines, Cal27, SCC4, and UM1, among
which the Cal27 cells showed the mostly differen-
tial expression (all p<0.05) (Figure 1B). Thus, Cal27
was selected in the following steps.
Silenced miR-135b-5p Sabotages TSCC Cell
Proliferation and Improves Apoptosis
miR-135b-5p was silenced in Cal27 cells, with the
results of RT-qPCR suggesting a successful trans-
fection (p<0.05) (Figure 2A). As it was observed
154 Analytical and Quantitative Cytopathology and Histopathology®
Guo et al
Figure 1 miR-135b-5p is highly expressed in TSCC tumor tissues and cells. (A–B) miR-135b-5p expression in TSCC tissues (A) and
TSCC cell lines (B) detected by RT-qPCR. Data are expressed as mean±standard deviation. The t test was used for pairwise comparison,
and one-way ANOVA was used to determine statistical significance. Tukey’s multiple comparisons test was applied for post hoc test.
**p<0.01. Repetition=3.
5. by CCK-8 method (Figure 2B) and flow cytometry
(Figure 2C–D), silenced miR-135b-5p could sup-
press TSCC cell proliferation, promote apoptosis,
and induce cell cycle arrest (all p<0.05).
miR-135b-5p Targets ARID1A
To further explore the mechanism of miR-135b-5p
in TSCC, a bioinformatics site (https:/
/cm.jefferson.
edu/rna22/Precomputed/) was employed to re-
veal that there were many gene-shared binding
sites with miR-135b-5p, such as ARID1A, PROX1,
FAM76A, SCMH1, and RUNX3. According to a
literature, ARID1A degradation led to a rapid de-
velopment of TSCC.19 Moreover, mice with con-
Volume 43, Number 4/August 2021 155
miR-135b-5p in TSCC Progression
Figure 2 Silenced miR-135b-5p sabotages TSCC cell proliferation and improves apoptosis. (A) miR-135b-5p expression in Cal27 cells
assessed using RT-qPCR. (B) Cal27 cell proliferation measured by CCK-8 method. (C–D) Cal27 apoptosis (C) and cell cycle (D) evaluated
through flow cytometry. Data are expressed as mean±standard deviation. The t test was used for pairwise comparison. **p<0.01.
Repetition=3.
6. ditional ARID1A knockout were susceptible to
TSCC or esophageal SCC.20 Therefore, ARID1A
was applied in dual luciferase reporter gene as-
say, from which we learned that as compared with
that transfected with mimic NC luciferase activi-
ty of ARID1A 3’UTR WT vector transfected with
miR-135b-5p mimic was significantly decreased,
while that of ARID1A 3’UTR MUT vector exhib-
ited no difference (p<0.05) (Figure 3A). On the
other hand, miR-135b-5p underexpression en-
hanced ARID1A expression in TSCC cells (p<0.05)
(Figure 3B). Then, as it was unveiled by RT-qPCR
and western blot analysis, ARID1A expression
was greatly quenched in TSCC tumor tissues
156 Analytical and Quantitative Cytopathology and Histopathology®
Guo et al
Figure 3 miR-135b-5p targets ARID1A in TSCC. (A) Binding site between miR-135b-5p and ARID1A 3’UTR as well as the luciferase
activity of ARID1A-WT and ARID1A-MUT co-transfected with mimic NC or miR-135b-5p mimic into 293T cells. (B) miR-135b-5p
expression in Cal27 cells detected by RT-qPCR and western blot analysis after silencing miR-135b-5p. (C–D) ARID1A expression in TSCC
tissues (C) and TSCC cell lines (D) evaluated by RT-qPCR and western blot analysis. Data are expressed as mean±standard deviation.
The t test was used for pairwise comparison, and one-way ANOVA was used to determine statistical significance. Tukey’s multiple
comparisons test was applied for post hoc test. *p<0.05, **p<0.01. Repetition=3.
7. and cells (all p<0.05) (Figure 3C–D). Simply put,
miR-135b-5p targeted ARID1A in TSCC.
ARID1A Knockdown Reverses the Inhibited TSCC
Cell Proliferation and Promoted Apoptosis Induced
by Silencing miR-135b-5p
miR-135b-5p and ARID1A were degraded in Cal27
cells, and RT-qPCR and western blot analysis
found that, compared with the miR-135b-5p in-
hibitor+si-NC group, the miR-135b-5p inhibitor+
si-ARID1A had declined ARID1A expression (p<
0.05) (Figure 4A).
According to CCK-8 method (Figure 4B) and
flow cytometry (Figure 4C), the miR-135b-5p in-
hibitor+si-ARID1A had elevated TSCC cell prolif-
eration and decreased apoptosis as compared with
the miR-135b-5p inhibitor+si-NC group (p<0.05).
Silenced miR-135b-5p Inactivates the PI3K/AKT Axis
by Regulating ARID1A
A previous research supported that the PI3K/AKT
axis activation encouraged gastric carcinoma cell
proliferation and invasion.21 It was also discov-
ered that the PI3K/AKT axis would by upregu-
lated by ARID1A knockout in pancreatic cancer.22
Western blot analysis was performed to assess the
PI3K/AKT axis activation of TSCC cells in each
group, and it found that silencing miR-135b-5p
contributed to the downregulation of the PI3K/
AKT axis; while ARID1A underexpression result
ed in activation of the PI3K/AKT axis (all p<0.05)
(Figure 5).
Discussion
TSCC represented one of the indomitable oral
cancers despite the clinical progress made during
the last few decades.23 It was discovered that miRs
with aberrant expression worked as tumor pro
motors or suppressors in oral tumorigenesis by
actively participating in cell activities such as dif
ferentiation, proliferation, survival, and death, in-
dicating their valuable role in oral carcinoma di-
agnosis and prognosis.24 miR-135b-5p experienced
an evident activation in individuals with ESCC.11
In the current study we discussed the mechanism
of miR-135b-5p and ARID1A in biological pro-
cesses of TSCC with the involvement of the PI3K/
AKT pathway. Consequently, we found silenced
miR-135b-5p blocked TSCC cell growth by nega-
tively modulating ARID1A and inactivating the
PI3K/AKT axis.
The first and foremost finding in this experi-
Volume 43, Number 4/August 2021 157
miR-135b-5p in TSCC Progression
Figure 4 ARID1A knockdown reverses the inhibited TSCC cell proliferation and promoted apoptosis induced by silencing miR-135b-5p.
(A) ARID1A expression in Cal27 cells measured using RT-qPCR and western blot analysis. (B) Cal27 cell proliferation determined by
CCK-8 method. (C) Cal27 cell apoptosis evaluated by flow cytometry. Data are expressed as mean±standard deviation. The t test was
used for pairwise comparison. **p<0.01. Repetition=3.
8. ment was that miR-135b-5p expression was ele-
vated in TSCC. Similarly, miR-135b-5p expressed
higher in tissues from patients with oral cancer
than those from the healthy volunteers.25 Highly
expressed miR-135b-5p in HNSCC motivated cell
colony formation and biological behaviors.15 On
the other hand, silenced miR-135b-5p quenched
TSCC proliferation, improved apoptosis, and in-
duced cell cycle arrest. In ESCC, miR-135b-5p ap-
peared to be related to the patients’ survival rate,
lymph node metastasis, and tumor differentia-
tion.11 miR-135b knockdown resulted in the slow-
down in cellular invasion and activity in CSCC.10
A previous literature documented that in SCCs,
the fact that apoptosis was strengthened was
beneficial to the repression of potential cancer de-
velopment and enhancement of tumor sensitiza-
tion to radiotherapy and chemotherapy.26 Besides,
downregulation of miR-135b encouraged the
growth of apoptosis factors,27 which indicated the
negative relation between miR-135b and apoptosis.
Importantly, we noticed that miR-135b-5p target-
ed ARID1A. ARID1A referred to a subunit gene
which was likely to mutate in different kinds of
cancers whose pathogenesis could be easily trig-
gered by the absence of ARID1A as this gene was
potent in maintaining enhancer activity and nor-
mal gene expression.28 ARID1A was reversely mod-
ulated by another oncogene in HNSCC so as to
amplify the stemness and carcinogenicity and con-
tribute to a poor survival rate of HNSCC.29 Like-
wise, as a target gene of a tumor promoter gene
in gastric cancer, ARID1A knockout could lead
to cancer gene invasion and growth.30 Besides,
ARID1A was able to enhance apoptosis to ensure
pancreatic cancer would be more sensitive to the
radiotherapy.22 That is to say, miR-135b-5p inhibi
tion was largely associated with different oral
malignancies with the involvement with ARID1A.
ARID1A knockdown reversed the sabotaged
TSCC cell proliferation and promoted apoptosis
caused by silencing miR-135b-5p, as revealed by
functional assays. As a famous and popular tumor
suppressor, ARID1A sabotaged cancer stem cells,
which was identified as the major cause of tumor
progression, and reduced the chemoresistance in
SCCs, thus generating a favorable prognosis.20 Put
in another way, stabilized expression of ARID1A
curbed SCC development and chemoresistance.19
It was reported that ARID1A loss brought about
growth of colorectal cancer via stimulating apop
tosis.31 In addition, silenced miR-135b-5p inacti-
vated the PI3K/AKT axis by negatively regulating
ARID1A. Data have supported that a lot of can-
cers, ranging from ovarian cancer to lung cancer,
witnessed AKT upregulation, which boosted tu-
158 Analytical and Quantitative Cytopathology and Histopathology®
Guo et al
Figure 5 Silenced miR-135b-5p inactivates the PI3K/AKT axis by regulating ARID1A. Protein levels of AKT, p-AKT, PI3K, and p-PI3K
of the PI3K/AKT axis in Cal27 cells were measured by western blot analysis. Data are expressed as mean±standard deviation. One-way
ANOVA was used to determine statistical significance. Tukey’s multiple comparisons test was applied for post hoc test. **p<0.01.
Repetition=3.
9. mor cell activity and mobility.32 According to a
recent study, the PI3K/AKT axis inhibitor con-
tributed to a better therapy for multiple cancers,
hinting at the detrimental role that the PI3K/
AKT axis played in human tumors.33 Importantly,
the PI3K/AKT axis and dysregulation of cell cycle
were both important players in ESCC progres-
sion.34 According to Jia et al, miR-135b directly
mediated the PI3K/AKT axis, an axis related to
tumor, to enhance colorectal cancer adhesion,
migration, and angiogenesis.35 Moreover, in pan-
creatic cancer with ARID1A knockout, the PI3K/
AKT axis was activated to reduce the efficiency of
regular radiotherapy.22
The main limitation of this study was that me
tastases and the prognosis of the 52 TSCC patients
were not analyzed. All in all, our study implied
that suppression of miR-135b-5p could suppress
TSCC cell growth by targeting ARID1A and inac-
tivating the PI3K/AKT pathway. These results are
promising in the prospect of promoting future al-
leviation of TSCC, thereby providing references for
optimizing individual treatment for patients with
TSCC. Though our findings offer therapeutic impli-
cation to TSCC therapy, the experimental results
and effective application into clinical practice still
need in-depth validation and exploration.
Ethical Approval
This study is approved by Ethics Committee of the
First Affiliated Hospital, Shihezi University School
of Medicine (2019-11-15,2019-089-01).
Consent for Publication
Informed consent was obtained from all individual
participants included in the study.
Availability of Data and Material
The datasets used or analyzed during the current
study are available from the corresponding author
on reasonable request.
Author Contributions
Guarantor of integrity of the entire study: Chao
Guo
Study concepts: Changxue Li
Study design: Chao Guo, Wenjing Yi
Definition of intellectual content: Chao Guo, Wen-
jing Yi, Changxue Li
Literature research: Wenjing Yi, Bin Sun, Xiaoyu
Zha
Clinical studies: Wenjing Yi, Xiaoxue Chen
Experimental studies: Chao Guo, Wenjing Yi, Bin
Sun, Xiaoyu Zha
Data acquisition: Xiaoxue Chen, Wenjing Yi
Data analysis: Xiaoyu Zha, Xiaoxue Chen
Statistical analysis: Bin Sun
Manuscript preparation: Chao Guo
Manuscript editing: Chao Guo, Zheng Zhou
Manuscript review: Zheng Zhou
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