Objective:To explore the effects of NOD2 gene on the proliferation and apoptosis of tongue squamous cell carcinoma Tca8113 cells.Methods:NOD2 expression vector(NOD2-pEZ-M29)and NOD2-shRNA vector were established,then were trans-fected into Tca8113 cells respectively.Expressions of HBD-2 and NOD2 in the cells was detected by RT-PCR and Western blot.Cell proliferation was examined by MTT assay and apoptosis by flow cytometry at 48h post transfection.Results:Compared with the control group,the expression of NOD2 and HBD-2 in NOD2-pEZ-M29 transfection group was significantly higher and markedly lower in NOD2-shRNA group.The proliferation rate of Tca8113 cells was markedly lower in NOD2-pEZ-M29 transfection group and signifi-cantly higher in NOD2-shRNA group while the apoptosis rate was significantly higher in NOD2-pEZ-M29 transfection group and sig-nificantly lower in NOD2-shRNA group.Conclusion:In Tca8113 cells NOD2 expression was positively correlated with HBD-2 ex-pression.NOD2 gene may promote the apoptosis,inhibit the proliferation of Tca8113 cells.

Objective:To develop a method for the detection of 10 single nucleotide polymorphisms(SNPs) of Nm23 gene,and to explore the genotypic and allelic distributions of the 10 SNPs in Chinese Hans population in Wuhan.Methods: Two hundreds healthy subjects ,115 men and 85 women included ,were enrolled as DNA sample donors.The real time TaqMan-MGB genotyping assay was used for the determination of the 10 SNPs selected ,and the results were validated by direct gene sequencing.Results:The method established could accurately and quickly screen the genotypes of the 10 SNPs of Nm23/NDPK gene.Distribution frequencies of the 10 SNPs were similar to these in other researches as well as these of HCB.All the loci follow the Hardy-Weinberg equilibrium.Highly linkage disequilibriums were found between rs 16949649 and rs7207370 , rs16949649 and rs34214448 , rs2159359 and rs2302254 , rs2159359 and rs8075231 ,rs2159359 and rs2041296 ,as well as rs2159359 and rs8071647 ,respectively.Four Tag SNPs:rs34214448 , rs2302254 ,rs11868380 and rs2318785 were initially selected by Heploview software.Conclusion:The method established for SNP gen-otyping can meet the needs for rapid analysis of Nm 23 gene polymorphisms ,and may have great values in investigating the association between gene polymorphisms and diseases as well as adverse drug reactions.

Objective:To establish a method for the detection of 8 single nucleotide polymorphisms (SNPs) of RAC1 gene,and to analyze the genotypic and allelic distributions of the 8 SNPs in healthy Chinese Han population.Methods:The real-time fluorescence TaqMan-MGB probes allele classification technology was used for the determination of the 8 SNPs in 150 cases of healthy volunteers in Hubei China ,and the results were validated by direct gene sequencing.Results: The method established in this study can accurately screen the genotypes of the 8 SNPs of human RAC1 gene.Highly linkage disequilibriums were found between rs 10951982 and rs9374 , rs702482 and 836488 ,respectively.All the 8 sits were in accordance with Hardy-Weinberg equilibrium.Six Tag-SNPs were selected by Heploview software:rs10951982 ,rs6954996 ,rs6951997 ,r1s2977 ,rs702482 and rs702483.The MAFs of the 8 SNPs in our study were close to the MAFs in CHB and Asian in Hap Map database .Significant distribution differences were also observed in different races.Conclusion:No significant differences are observed in this study in healthy Chinese Han population.But differences are found when compared with the data of other races in Hap Map.

AIM:Mitochondrial DNA (mtDNA) copy number variation (CNV), which reflects the oxidant-induced cell damage, has been observed in a wide range of human diseases .However, whether it correlates with heart failure , which is closely related to oxidative stress, has never been elucidated before .We aimed to systematically investigate the association between leukocyte mtDNA CNV and heart failure risk and prognosis .METHODS: A total of 1 700 hospitalized patients with heart failure and 1 700 age-and gender-matched community population were consecutively enrolled in this observational study , as well as 1 638 ( 96.4%) patients were fol-lowed prospectively for a median of 17 months (12~24 months).The relative mtDNA copy number in leukocyte of peripheral blood or cardiac tissue was measured in triplicate by quantitative real-time PCR method .RESULTS:Patients with heart failure possessed much lower relative mtDNA copy number compared with control subjects (P<0.01), especially for the patients with ischemic etiology (P<0.01).Patients with lower mtDNA copy number exhibited 1.7 times higher risk of heart failure ( P<0.01).Long-term follow-up (median 17 months) showed that decreased mtDNA copy number was significant associated with both increased cardiovascular deaths (P<0.01) and cardiovascular rehospitalization (P<0.01).After adjusted for the conventional risk factors and medications , lower mtDNA copy number were still significantly associated with 50% higher cardiovascular mortality (P <0.05).CONCLUSION:
mtDNA copy number depletion is an independent risk factor for heart failure and predicted higher risk of cardiovascular deaths in patients with heart failure .

Background: The onset and progression of type 1 diabetes mellitus (T1DM) is closely related to autoimmunity. Effective monitoring of the immune system and developing targeted therapies are frontier fields in T1DM treatment. Currently, the most available tissue that reflects the immune system is peripheral blood mononuclear cells (PBMCs). Thus, the aim of this study was to identify key PBMC biomarkers of T1DM. Methods: Common differentially expressed genes (DEGs) were screened from the Gene Expression Omnibus (GEO) datasets GSE9006, GSE72377, and GSE55098, and PBMC mRNA expression in T1DM patients was compared with that in healthy participants by GEO2R. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and protein-protein interaction (PPI) network analyses of DEGs were performed using the Cytoscape, DAVID, and STRING databases. The vital hub genes were validated by reverse transcription-polymerase chain reaction using clinical samples. The disease-gene-drug interaction network was built using the Comparative Toxicogenomics Database (CTD) and Drug Gene Interaction Database (DGIdb). Results: We found that various biological functions or pathways related to the immune system and glucose metabolism changed in PBMCs from T1DM patients. In the PPI network, the DEGs of module 1 were significantly enriched in processes including inflammatory and immune responses and in pathways of proteoglycans in cancer. Moreover, we focused on four vital hub genes, namely, chitinase-3-like protein 1 (CHI3L1), C-X-C motif chemokine ligand 1 (CXCL1), matrix metallopeptidase 9 (MMP9), and granzyme B (GZMB), and confirmed them in clinical PBMC samples. Furthermore, the disease-gene-drug interaction network revealed the potential of key genes as reference markers in T1DM. Conclusion These results provide new insight into T1DM pathogenesis and novel biomarkers that could be widely representative reference indicators or potential therapeutic targets for clinical applications.

Objective: : Ischemia and hemorrhage of pituitary adenomas (PA) caused important clinical syndrome. However, the differences on clinical characteristics and surgical outcomes between these two kinds apoplexy were less reported. Methods: : A retrospective analysis was made of patients with pituitary apoplexy between January 2013 and June 2018. Baseline and clinical characteristics before surgery were reviewed. All patients underwent transsphenoidal surgery and were followed up at least 1 year. Results: : Total 67 cases (5.8%) among 1147 pituitary tumor patients were enrolled, which consisted of 28 (~2.4%) ischemic PA and 39 (~3.4%) hemorrhagic PA. There were more male patients in the ischemic group compared with hemorrhagic group (78.6% vs 53.8%, p=0.043). However, the mean age, tumor size and functional tumor ratio were significant higher in the hemorrhagic group. Headache was more common in ischemic PA (82.1%) than that of hemorrhagic PA (51.3%, p=0.011). Magnetic resonance imaging findings found that mucosal thickening and enhancement of the sphenoid sinus was observed in 15 ischemic PA patients (n=27, 55.6%), but none in patients with hemorrhagic PA (n=38, p<0.0001). It was worth noting that the rate of pre-surgical hypopituitarism in ischemic PA patients were seemed higher than that in hemorrhagic PA patients, but not significant. The two groups got a total tumor resection rate at 94.1% and 92.9%, independently. No significant difference on the operative time, blood loss in operation and complications in perioperative period was observed in two groups. After operation, cranial nerve symptoms recovered to normal at 81.8% of ischemic PA patients and 82.6% of hemorrhagic PA patients. Importantly, the incidence of postoperative hypopituitarism partially decreased in both groups, among which the rate of hypothyroidism in ischemic PA patients significantly decreased from 46.4% to 18.5% (p=0.044). Conclusion : Patients with ischemic PA presented different clinical characteristics to the hemorrhagic ones. Transsphenoidal surgery should be considered for the patients with neuro-ophthalmic deficits and might benefit for pituitary function recovery of the apoplectic adenoma patients, especially pituitary thyroid axis in ischemic PA patients.

Objective:To observe the effects of glucagon-like peptide-1 receptor agonist(GLP-1RA) combined with metformin on glucolipid metabolism and reproductive function in overweight/obesity polycystic ovarian syndrome(PCOS) patients.Methods:Retrospectively analyzed changes in clinical parameters of body measurements, glucolipid metabolism, menstrual cycle, hormones, and polycystic ovary in 200 overweight/obese PCOS Patients who received 12 weeks of treatment(liraglutide+ metformin or exenatide+ metformin) in the Department of Endocrinology at the Second Affiliated Hospital of Army Medical University from July 2017 to July 2022.Results:In terms of metabolism improvement, body weight, body mass index, waist circumference, hip circumference, waist-hip ratio, glutamic-oxalacetic transaminase, γ-glutamyltransferase, glycosylated hemoglobin, fasting blood glucose, insulin(including fasting, 30, 60, 120, and 180 minutes), homeostasis model assessment for insulin resistance, area under curve-insulin, triglyceride, and total cholesterol were significantly decreased after treatment( P<0.05 or P<0.01 or P<0.001). In terms of reproductive function, testosterone, luteinizing hormone, follicle stimulating hormone/luteinizing hormone and free androgen index were decreased( P<0.001), while sex hormone-binding globulin was increased( P<0.01). There were no significant differences in progesterone, prolactin, follicle-stimulating hormone and dehydroepiandrosterone sulfate compared with before treatment( P>0.05). The proportion of subjects with regular menstrual cycle increased from 23.53% to 57.52%( P<0.05). The proportion of subjects with polycystic ovarian changes decreased from 65.30% to 50.32%, and the proportion with dominant follicles increased from zero to 18.30%( P<0.05 or P<0.01). Some patients(25.49%) experienced adverse reactions such as nausea, diarrhea, abdominal distension, and vomiting after medication. Conclusion:The combination of GLP-1RA and metformin effectively improves glucose lipid metabolism disorder and reproductive dysfunction in overweight/obese PCOS patients.