Aberrant histone modification and inflammatory cytokine production of peripheral CD4+ T cells in patients with oral lichen planus.

BACKGROUNDS: To investigate alterations in histone modification and histone deacetylases (HDACs) in patients with oral lichen planus (OLP), and to evaluate correlations with inflammatory cytokine production. METHODS: Global histone H3/H4 acetylation and HDAC activity in CD4+ T cells from 23 patients with OLP and 10 healthy control subjects were examined using spectrophotometry. The mRNA levels of eight members of four classes of HDAC genes were measured by real-time quantitative polymerase chain reaction. Forty cytokines involved in inflammation were examined with a cytokine array. The correlation between histone modification and cytokine production was analyzed. RESULTS: Global histone H3 hypo-acetylation was observed in OLP patients. Patients with OLP had significantly higher HDACs activity,and higher HDAC6 and HDAC7 mRNA level compared with the controls. Of the 40 cytokines in the cytokine array, eight were significantly increased in OLP patients: interleukin (IL)-4, IL-8, IL-1ra, tumor necrosis factor receptor II (TNFR II), macrophage inflammatory protein 1b (MIP-1b), fibrosis-associated tissue inhibitors of metalloproteinase 1 (TIMP)-1, monocyte chemotactic protein 1 (MCP-1), and eotaxin-2. In the OLP group, the acetylation level of histone H3 was negatively correlated with IL-4 and MCP-1 production, and the expression of HDAC6 mRNA was positively correlated with MCP-1 production. In the non-erosive subgroup, acetylation of histone H3 was negatively correlated with IL-4, IL-16, and TIMP-2 production. In the erosive OLP subgroup, the expression of HDAC7 mRNA was positively correlated with MIP-1a production. CONCLUSION: Aberrant histone modification of CD4+ T cells in peripheral blood could occur in OLP patients, and possibly affects inflammatory cytokine production.

Effects of PTEN gene silencing on invasion and EMT in oral squamous carcinoma Tca8113 cells.

AIM: To investigate the impact of silencing of the PTEN gene using siRNA on the invasion, proliferation, cell cycle, and epithelial-mesenchymal transition of the Tca8113 cell line. METHODS: The established Tca8113 cell model with siRNA interference to silence the PTEN gene was used. The transfection efficiency was examined by RT-qPCR and Western blot analysis. CCK-8 assay was utilized to analyze the proliferation of Tca8113 cells and cell invasion was evaluated using a transwell assay. The cell cycle distribution was analyzed by flow cytometry. The protein expression levels of the epithelial-mesenchymal transition (EMT) markers E-cadherin and Vimentin and the EMT-related proteins ß-catenin and TGF-ß1 were analyzed by Western blot. RESULTS: The expression level of PTEN was significantly reduced in the PTEN-siRNA group. The invasiveness and proliferation rate of Tca8113 cells in the PTEN-siRNA group were significantly greater than those of the control and negative control groups. The expression levels of E-cadherin and ß-catenin were reduced, whereas the expression levels of vimentin and TGFß-1 were elevated in the PTEN-siRNA group compared with those of control and negative groups. These results were significantly different. CONCLUSION: The silencing of PTEN by siRNA increased the proliferation and promoted cell invasion of Tca8113 cells. PTEN gene silencing may accelerate the EMT in Tca8113 cells.

Upregulation of PTEN suppresses invasion in Tca8113 tongue cancer cells through repression of epithelial-mesenchymal transition (EMT).

We previously discovered that the expression of the tumor suppressor phosphatase and tensin homolog (PTEN) was downregulated in the majority patients with tongue squamous cell carcinoma (TSCC). The aim of this study was to investigate the role of PTEN overexpression in the regulation of epithelial-mesenchymal transition (EMT) of the tongue squamous carcinoma cell line Tca8113 as well as explore the underlying mechanism. GV230 (containing the PTEN gene) and empty vectors were transfected into Tca8113 cells. After stable transfection, the messenger RNA (mRNA) and protein levels of PTEN were validated using quantitative real-time PCR (qPCR) and Western blot analysis. The growth and cell cycle were analyzed using Cell Counting Kit-8 (CCK-8) and flow cytometry, respectively. The invasion ability was measured with a transwell assay. The effects of PTEN overexpression on EMT and Hedgehog signaling were assessed by comparing Tca8113-PTEN cells with control and negative control cell groups. We found that PTEN expression was significantly upregulated after transfection. Meanwhile, upregulated PTEN inhibited the proliferation and invasion of Tca8113 cells. In addition, we observed changes in the EMT- and Hedgehog-associated proteins. These data demonstrated that PTEN upregulation could reduce invasion by inhibiting the process of EMT in Tca8113 cells, which might be related to the Hedgehog signaling pathway.

Effect of comprehensive nursing intervention on the outcomes of in vitro fertilization in patients with polycystic ovary syndrome: A randomized controlled study.

OBJECTIVE: To explore the effects of comprehensive nursing intervention on in vitro fertilization (IVF) and pregnancy outcomes in patients with polycystic ovary syndrome (PCOS). METHOD: A total of 130 patients with PCOS admitted to our hospital from April 2021 to March 2023 were selected as the research subjects. They were evenly divided according to a random number table method. The control group received routine care for the patients, while the study group received comprehensive care for the patients. The IVF, pregnancy outcomes, negative emotional changes, serum and follicular fluid (FF) amyloid-related protein and C-reactive protein (CRP) levels of the 2 groups of patients were compared. RESULT: The data on IVF rate and pregnancy rate in the study group were significantly better than those in the control group (P < .05). The SAS and SDS scores of the study group patients after intervention were significantly lower than those of the control group (P < .05). After intervention, the levels of serum and FF amyloid associated protein and CRP in the study group were significantly lower than those in the control group (P < .05). CONCLUSION: Patients with PCOS who receive comprehensive care can increase their probability of IVF, improve their pregnancy outcomes, and have a positive significance in reducing negative emotions.

Novel susceptibility genes were found in a targeted sequencing of stroke patients with or without depression in the Chinese Han population.

BACKGROUND: Both stroke and depression are multi-factorial diseases, with both genetic and environmental factors likely to participate in their pathogenesis. Post stroke depression (PSD) is a common complication after stroke leading to poor functional outcome, increased physical disability and mortality. Although several genes have been associated with PSD, the genetic basis of PSD remains poorly understood. METHOD: A 2-stage candidate gene study by targeted sequencing was conducted involving stroke patients with or without depression and health controls. In the discovery stage (121 PSD, 131 non-PSD and 639 HC), logistic regression was used to test associations respectively in PSD and non-PSD groups. In the replication stage (200 PSD, 218 non-PSD and 983 HC), 54 selected SNPs were again genotyped in an independent cohort. Fixed-effects inverse variance-weighted meta-analysis was used in the combined samples. RESULTS: The study identified 2 novel genes associated with PSD [HTR3D (rs55674402, p = 0.002512, odds ratio (OR) = 0.7431); NEUROG3 (rs144643855, p = 0.00325, OR = 0.6523)] and 3 risk SNPs in one risk gene associated with non-PSD [PIK3C2B (rs17406271, p = 0.0006801, OR = 1.446; rs2271419, p = 0.0005836, OR = 1.497; rs2271420, p = 0.001031, OR = 1.431)] in the Chinese sample. NEUROG3 shows highest expression level in hippocampus. Functional enrichment analysis shows that susceptibility genes for PSD are mostly enriched in chemical synaptic transmission and regulation of lipid synthetic process. LIMITATIONS: The sample size was not sufficient to reach a genome-wide p value level. To overcome this shortage, some unique strategies were applied during the selection of SNPs for replication. Secondly, the age, gender composition and depressive severity between two stages were not well-matched. Different sample sources should be blamed, and to minimizing the influence, gender was corrected as co-variant in logistic regression. CONCLUSION: This study identified that HTR3D and NEUROG3 were linked with the susceptibility of PSD and PIK3C2B with stroke in the Chinese Han population. Further replication of these findings in a larger and better matched sample is warranted. Functional analysis suggests that the pathogenesis of PSD may be implicated in 5-HT synaptic transmission, neural plasticity and lipid metabolism, and therapeutic interventions targeting these pathways may be effective approaches for PSD treatment.

[Expressions of PTEN and FHIT in oral squamous cell carcinoma and their relations with cyclin D1].

OBJECTIVE: To study the expressions of PTEN, FHIT and cyclin D1 in normal oral mucosa and oral squamous cell carcinoma (OSCC), and analyze the relationship between PTEN/FHIT and cyclin D1. METHODS: Immunohistochemical staining with SP methods was used to detect the expression of PTEN, FHIT and cyclin D1 in OSCC tissues in 62 cases and normal oral mucosa in 12 cases. RESULTS: The positivity rates of PTEN and FHIT were both 100% (12/12) in the normal oral mucosa, while in 62 cases of OSCC, 24.2% (15/62) and 17.7% (11/62) were negative for (or with low expression of) PTEN and FHIT, respectively. In normal oral mucosa, 91.7% (11/12) cases had negative or low cyclin D1 expression, while 53.2% (33/62) of the OSCC cases were positive for cyclin D1 expression. In all the cases, when PTEN and FHIT were strongly expressed, 37.8% (28/74) of the cases had negative or low expression of cyclin D1, including 11 normal cases. CONCLUSION: The tumor suppressor genes PTEN and FHIT play a role in the pathogenesis of OSCC, and PTEN and FHIT can down-regulate the expression of cyclin D1.

[Expression and significance of fragile histidine triad in oral cancer and precancerous lesions].

OBJECTIVE: To detect the expression of fragile histidine triad (FHIT) in oral cancer and oral precancerous lesions and investigate the relationship between the FHIT expression and the histopathological changes. METHODS: Immunohistochemical staining by SP methods was utilized to detect the expression of FHIT in 64 cases of oral squamous cell carcinoma, 39 oral precancerous lesions and 12 normal oral mucosa specimens. RESULTS: The positivity rate of FHIT in normal oral membrane was 100% (12/12), and medium and high FHIT expression levels were detected in oral precancerous lesions but without significant difference from that in normal oral mucosa. In 64 oral squamous cell carcinoma specimens, 3 were negative for FHIT expression, 8 had low expression, and the other 43 had moderate and high expression. The rate of negative or low FHIT expression in the oral squamous cell carcinoma was 17% (11/64), which was significantly different from that in the normal oral mucosa and oral precancerous lesions, but the rate was not correlated to the differentiation of the cancer. CONCLUSION: The tumor suppressor gene FHIT plays a role in the pathogenesis of oral squamous cell carcinoma.

[Expressions of FHIT and cyclin D1/CDK4 in oral cancer and oral precancerous lesions].

OBJECTIVE: To detect the expressions of fragile histidine triad (FHIT) and cyclin D1/CDK4 in oral squamous cell carcinoma (OSCC) and oral precancerous lesions and investigate the relationship between the expressions and the histopathological changes. METHODS: Immunohistochemical staining by SP methods was utilized to detect the expression of FHIT and cyclin D1/CDK4 in 64 cases of OSCC, 39 oral precancerous lesions and 12 normal oral mucosa specimens. RESULTS: The rate of the negative or low FHIT expression in OSCC was 17% (11/64), which was remarkably lower than that in normal oral membrane and oral precancerous lesions (P<0.01). Significantly higher levels of cyclin D1/CDK4 expressed in OSCC than in normal oral membrane and precancerous lesions (P<0.01). There was no significant correlation between FHIT and cyclin D1/CDK4, and positive correlation between cyclin D1 and CDK4 was observed. CONCLUSION: FHIT, cyclin D1 and CDK4 may play a role in the pathogenesis of OSCC, and FHIT can down-regulate the expression of cyclin D1.

[The expression of E-cadherin, vimentin, ß-catenin, transforming growth factor-ß1 in oral squamous cell carcinomas].

OBJECTIVE: To investigate the expression of E-cadherin, vimentin, ß-catenin and transforming growth factor-ß1 (TGF-ß1) in oral squamous cell carcinomas (OSCC). METHODS: Eighty-nine cases of OSCC and 20 cases of normal oral mucosa were collected. Then the 89 cases of OSCC were classified as grade I, II, III. The semiquantitative method was used to calculated the positive intensity and positive rate. The relationship between the OSCC differentiation and the four biomarkers was analyzed. RESULTS: The median of E-cadherin was 9.00 in the normal tissue, 9.00, 6.00 and 6.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-4.211, P=0.000). The median of vimentin was 0.00 in the normal tissue, 0.00, 0.00 and 4.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-3.675, P=0.000). The median of ß-catenin was 9.00 in the normal tissue, 3.00, 4.00 and 3.00 in OSCC I, II and III, respectively. There was significant difference between the normal group and OSCC group (Z=-6.300, respectively. There was significant difference between the normal group and OSCC group (Z=-3.329, P=0.000). E-cadherin expression was positively correlated to ß-catenin expression (r=0.327, P=0.002), negtively correlated to vimentin expression (r=-0.386, P=0.001) and positively correlated to TGF-ß1 expression (r=-0.304, P=0.004). Vimentin expression was positively correlated to TGF-ß1 expression (r=0.401, P=0.000). CONCLUSIONS: E-cadherin and ß-catenin in OSCC had a down-regulated expression, while the vimentin has an up-regulated expression.

[Expression of caspase-8, receptor interacting protein and nuclear factor-kappaBp65 in oral lichen planus].

OBJECTIVE: To investigate the expressions of caspase-8, receptor interacting protein (RIP) and nuclear factor (NF)-kappaBp65 in oral lichen planus (OLP) and their relationship with cell apoptosis. METHODS: Immunohistochemical technique with SP method was used to detect the expressions of caspase-8, RIP and NF-kappaBp65 in 30 OLP cases and 15 normal oral mucosa specimens. Terminal deoxynucleotidyl transferase-mediated nucleotide shift enzyme (TdT) mediated d-UTP end labeling (TUNEL) was used for detecting the cell apoptotic index (AI) in 15 OLP cases and 5 nomal oral mucosa specimens. RESULTS: Compared with the control group, the AI of epithelial cells (6.76 +/- 2.32) increased and the AI of lymphocytes (1.75 +/- 0.74)decreased in OLP (P < 0.01). The positive rate of caspase-8, RIP and NF-kappaBp65 of epithelial cells were 97% (29/30), 87% (26/30) and 93% (28/30) respectively, significantly higher in OLP than in normal control (P < 0.05). The positive rate of caspase-8, RIP and NF-kappaBp65 of lymphocytes were 100% (30/30, 90% (27/30) and 80% (24/30) respectively, significantly higher in OLP than in normal control (P < 0.01). A positive correlation was also observed between NF-kappaBp65 expression of lymphocytes and AI of epithelial cells. CONCLUSIONS: Accelerated apoptosis of the keratinocytes and inhibition of lymphocyte apoptosis may coexist and contribute to the formation and progression of OLP. The over expression of caspase-8, RIP and NF-kappaBp65 in OLP may play a role in the pathogenesis of OLP.

[Expressions of receptor-interacting protein and caspase-8 in oral squamous cell carcinoma and oral precancerous lesions].

OBJECTIVE: To detect the expressions of receptor-interacting protein (RIP) and caspase-8 and investigate their roles in oral squamous cell carcinoma (OSCC) and oral precancerous lesions. METHODS: SABC immunohistochemical methods were used to detect the expressions of RIP and caspase-8 in 22 specimens of OSCC, 14 specimens of oral lichen planus (OLP), 14 specimens of oral leukoplakia (OLK) and 10 specimens of normal oral mucosa (NOM). RESULTS: The rate of weak or negative expression of RIP in normal mucosa was 50% (5/10). The rates of weak and positive expression of RIP in OLP, OLK and OSCC were 75% (36/50), and the rate of positive and strong expression of RIP was 63.7% (14/22) in OSCC, significantly higher that in the others groups (P<0.05). The rates of weak, positive and strong positive expression of caspase-8 in NOM, OLP, OLK and OSCC were 80% (8/10), 100% (14/14), 85.7% (12/14), and 100% (22/22), respectively. CONCLUSION: Both RIP and caspase-8 may play important roles in the occurrence and progression of OSCC and oral precancerous lesions.

[Expressions of NF--kappaBp65, TRAF2, cyclinD1 and their association with cell apoptosis in oral lichen planus].

OBJECTIVE: To examine the expression and distribution of NF-kappaBp65, TRAF2, and cyclinD1 and their association with cell apoptosis in oral lichen planus (OLP). METHODS: Sixty OLP patients were divided into erosion-atrophy group (n=30) and non-erosion group (n=30) according to their clinical features. Immunohistochemistry with SP method was used to detect the expressions of NF-kappaBp65, TRAF2, cyclinD1 in the 60 OLP and 40 normal oral mucosa (control) specimens. TUNEL assay of randomly selected specimens from 10 normal and 15 OLP cases was performed to detect the cell apoptotic index (AI). RESULTS: Compared with the control group, OLP group showed significantly increased AI of the epithelial cells (67.32-/+18.99) and decreased AI of the lymphocytes (34.12-/+9.89) (P<0.05). In the OLP group, the positivity rates for NF-kappaBp65, TRAF2, and cyclin D1 in the epithelial cells (85.00%, 76.67% and 71.67%, respectively) and in the lymphocytes (91.67%, 86.67% and 70.00%, respectively) were all significantly higher than those in the control group (P<0.05). NF-kappaBp65 expression was significantly increased in the lamina propria in the non-erosion OLP group as compared to the erosion-atrophy group. A positive correlation was noted between lymphocyte NF-kappaBp65 expression and AI of the epithelial cells, but an inverse correlation found between lymphocyte NF-kappaBp65 expression and the AI of the lymphocytes. Lymphocyte TRAF2 and cyclin D1expressions were also inversely correlated to lymphocyte AI. There was a positive correlation between TRAF2 and cyclin D1 expressions and the expression NF-kappaBp65 in the epithelial cells and lymphocytes in OLP. CONCLUSIONS: Accelerated apoptosis of the keratinocytes and inhibition of lymphocyte apoptosis may coexist to contribute to the formation and progression of OLP. NF-kappaBp65 expression, particularly its abnormal nuclear expression, may play a partial role in the pathogenesis of OLP.

[Expression of tumor suppressor gene PTEN, PIP3 and cyclin D1 in oral squamous cell carcinoma and their correlations].

OBJECTIVE: To detect the expression of PTEN, PIP3 and cyclin D1 in oral squamous cell carcinoma and precancerous lesions and analyze their correlation. METHODS: Immunohistochemistry SP method was used to detect the expression of PTEN, PIP3 and cyclin D1 in 63 cases of oral squamous cell carcinoma, 29 cases of simple hyperplasia, 33 cases of dysplasia, and 25 cases of normal oral mucosa. RESULTS: The negative or low expression of PTEN in oral squamous cell carcinoma was 25%, which was remarkably lower than that in other groups. The positive expression of PIP3 in simple hyperplasia, dysplasia and oral squamous cell carcinoma was 66%, 64%, and 76% respectively, which were much higher than those in normal oral mucosa. The positive expression of cyclin D1 in oral squamous cell carcinoma was 49%, which was significantly higher than that in other groups. The negative correlation between PTEN with PIP3, cyclin D1 and the positive correlation between PIP3 and cyclin D1 were observed. CONCLUSIONS: PTEN may play a role in the oncogenesis of oral squamous cell carcinoma, and PTEN may down-regulate the expression of PIP3, and then down-regulate the expression of cyclin D1, which leads to the suppression of cell growth.