Overexpression of miR-125 inhibits proliferation and invasion of nasopharyngeal carcinoma cells by down-regulating FGF-2 / 国际肿瘤学杂志

Objective:To explore the expression of miR-125 in nasopharyngeal carcinoma tissues and the possible regulatory mechanism of biological characteristics of tumor cells.Methods:Thirty cases of carcinoma and paracancerous tissues from patients with nasopharyngeal carcinoma admitted to Southern Hospital of Sixth People′s Hospital Affiliated to Shanghai Jiaotong University from June 2018 to June 2019 were collected. The expressions of miR-125 and fibroblast growth factor 2 (FGF-2) mRNA were detected by fluorescent quantitative PCR. Nasopharyngeal carcinoma CNE-2 cells were transfected by miR-125 mimic (miR-125 mimic group), and negative control group was set (NC group). Transwell chamber assay was used to determine cell invasion ability, scratch healing assay was used to determine cell migration ability, WST-1 assay was used to assess cell viability, flow cytometry and electron microscopy were used respectively to detect apoptosis and autophagy, dual luciferase reporter assay was used to analyze the target of miR-125, and Western blotting was used to detect the expressions of related proteins.Results:The relative expression of miR-125 mRNA in nasopharyngeal carcinoma tissues was 0.692±0.316, which was significantly lower than 1.501±0.748 in the adjacent tissues ( t=5.242, P<0.001). The relative expression of FGF-2 mRNA in nasopharyngeal carcinoma tissues was 1.317±0.552, which was significantly higher than 0.783±0.241 in the adjacent tissues ( t=7.360, P<0.001). The miR-125 mRNA expression of CNE-2 cells in the miR-125 mimic group was 4.091±0.145, which was significantly higher than 0.993±0.137 in the NC group ( t=85.062, P<0.001). The proliferative activities of CNE-2 cells in the miR-125 mimic group at 48 and 96 h after transfection were significantly lower than those in the NC group (0.891±0.214 vs. 1.295±0.245, t=6.802, P<0.001; 0.934±0.208 vs. 1.488±0.269, t=8.924, P<0.001). The number of transmembrane cells and cell migration rate of the miR-125 mimic group were 36 000±3 820 and (39.4±6.5)%, which were significantly lower than 74 000±7 500 and (102.7±10.6)% of the NC group ( t=24.728, P<0.001; t=27.883, P<0.001). The apoptosis rate of CNE-2 cells in the miR-125 mimic group was (22.5±1.4)%, which was significantly higher than that in the NC group (1.4±0.5)% ( t=77.740, P<0.001). The relative expression of the apoptotic protein Bax in the miR-125 mimic group was 0.983±0.158, which was significantly higher than that in the NC group (0.418±0.122; t=15.503, P<0.001), and the relative expression of Bcl-2 was 0.688±0.174, which was significantly lower than that of the NC group (1.013±0.109; t=8.670, P<0.001). Autophagy was observed in miR-125 overexpressing CNE-2 cells by electron microscopy, and the relative expression ratio of autophagy protein LC3Ⅱ/LC3Ⅰ in the miR-125 mimic group was 2.517±0.209, which was significantly higher than 1.238±0.135 in the NC group ( t=28.156, P<0.001). The expression of FGF-2 protein in the miR-125 mimic group was 0.504±0.118, which was significantly lower than 1.228±0.134 in the NC group ( t=22.210, P<0.001). The double luciferase report confirmed FGF-2 as the target gene of miR-125. At 12, 24, 48 and 96 h after the transfection, the cell proliferative activities of CNE-2 cells co-transfected by FGF-2 gene plasmid and miR-125 mimic were significantly higher than those of CNE-2 cells transfected by miR-125 mimic (all P<0.05), and the apoptosis rate was significantly lower than that of CNE-2 cells transfected by miR-125 mimic [(6.2±1.5)% vs. (17.6±2.4)%, t=22.062, P<0.001]. Conclusion:The expression of miR-125 is down-regulated in nasopharyngeal carcinoma tissues. Overexpression of miR-125 may inhibit the proliferation, migration and invasion of nasopharyngeal carcinoma CNE-2 cells and promote the apoptosis and autophagy of CNE-2 cells by down-regulating FGF-2 expression.

Formula Optimization of Aprepitant Nanocapsules by Central Composite Design-response Surface Method-ology / 中国药师

Objective:To optimize the formula of aprepitant nanocapsules. Methods: The central composite design–response surface methodology was used. The amounts of hydroxg propyl cellulose(HPC-SL) and sodium dodecyl sulfate(SDS) were set as the independent variables;the dissolution of aprepitant capsules at 15min and 30min, and the dissolution after accelerated at 40℃ and 75% RH for 6 months were set as the dependent variables. Quadratic polynomial mathematic models were used to evaluate the relation-ship between the independent and the dependent variables. According to the mathematic models,an effect graph was drawn. The opti-mized formula was chosen from the overlap of the contour graphs of the dependent variables. The similarity of in vitro dissolution curve was evaluated by using f2factor. Results:The correlation coefficient of quadratic polynomial mathematic model and the reliability was high. The measured values of the optimized formula were within the expected ranges. Conclusion: Aprepitant nanocapsules with the optimized formula by central composite design-response surface methodology meet the requirements. The results can provide evidence for the next industrial production.

Telmisartan reduces retina vessel endothelial cell apoptosis via upregulating retinal ACE2-Ang-(1-7)-Mas axis in spontaneous hypertensive rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the effects of angiotensin II (Ang II) antagonist telmisartan on retina vessel endothelial cell apoptosis and its impact on the ACE2-Ang-(1-7)-Mas axis in spontaneous hypertensive rats (SHR).</p><p><b>METHODS</b>Thirty-six SHR 16 week-old were randomly divided into 3 groups (n = 12 each): SHR, SHRT (telmisartan 10 mg · kg-1 · d-1 by gastric gavage) and SHRTA group (telmisartan 10 mg · kg-1 · d-1 by gastric gavage plus intravenous injection of A-779 0.5 mg · kg-1 · d-1), twelve WKY rats served as normotensive control group. Systolic blood pressure was measured at pre-treatment and 8 weeks later. After 8 weeks, rats were sacrificed, the expression of ACE2 and Mas in retina were analyzed by qRT-PCR, Western blot and Immunohistochemistry, the Ang-(1-7) concentration in serum was measured by ELISA. Specimens were obtained and stained by hematoxylin and eosin, and the morphology of retina vessel was observed. Apoptosis of vessel endothelial cells were determined by using terminal deoxynucleotidyl transferase mediated dUTP nick end labeling method.</p><p><b>RESULTS</b>The systolic blood pressure of SHR, SHRT and SHRTA groups at baseline were significantly higher than age-matched WKY group (all P < 0.01). Eight weeks later, the systolic blood pressure group was significantly lower in SHRT group than in the SHR group (P < 0.01), this effect was partly reversed in SHRTA group. The retinal ACE2 mRNA and protein expression was significantly lower in SHR group than in WKY and SHRT groups (P < 0.01), which was similar between SHRT group and SHRTA group (P > 0.05). The retinal Mas mRNA and protein expression were significantly lower in SHR group compared to WKY and SHRT groups (all P < 0.01), which was significantly lower in SHRTA group than in the SHRT group (P < 0.05). ELISA results showed that serum Ang-(1-7) protein level was significantly lower in SHR group than in WKY group and SHRT group (both P < 0.05), which was lower in SHRTA group compared to SHRT group. Retinal vessel endothelial cell apoptosis was higher in SHR group than in WKY group, which could be reduced by cotreatment with telmisartan and this beneficial effect could be reversed by A-779.</p><p><b>CONCLUSION</b>Telmisartan can reduce retinal vessel endothelial cell apoptosis via upregulating the ACE2-Ang-(1-7)-Mas axis.</p>

Application of Mimics three-dimensional imaging in percutaneous left atrial appendage closure with Watchman system / 中华心血管病杂志

<p><b>OBJECTIVE</b>The three-dimensional (3D) structure of left atrial appendage (LAA) in atrial fibrillation patients were reconstructed by Mimics 3D imaging system, aiming at guiding for selection of both the size and location of the closure devices and making preliminary risk assessment of LAA closure with Watchman system.</p><p><b>METHODS</b>Inclusion criteria were: ten voluntary patients with both atrial fibrillation and indication for LAA closure aging from 40 to 85 years old with contraindication for oral anticoagulants or unwillingness to take long-term oral anticoagulation therapy from May to December 2014. 3D reconstruction of LAA was preoperatively made by Mimics 3D imaging system. With the Mimics 3D reconstruction model and the results of both transesophageal echocardiography (TEE) and LAA radiography, the size and location for the closure device were chosen. The devices were planted at the ostium of the LAA.</p><p><b>RESULTS</b>Ten atrial fibrillation patients were enrolled (average age: (66.3±11.9) years old) and all successfully implanted with the Watchman LAA closure devices. Nine of them were with non-valvular atrial fibrillation with average CHADS2-VAS score (3.2±1.7) and HAS-BLED score (2.7±1.6). The rest one was a valvular atrial fibrillation patient with the history of the percutaneous balloon mitral valvuloplasty (PBMV) without surgical indications of mitral valve replacement (MVR). There was no blood leakage around the device by regular postoperative TEE and LAA radiography examinations. There were no complications of bleeding, embolism, or stroke through both at peri-operative period and at 1 month follow-up post procedure.</p><p><b>CONCLUSION</b>Preoperative Mimics 3D reconstruction of LAA by Mimics 3D imaging system among atrial fibrillation patients provides essential information guiding the successful LAA closures.</p>

Association of GIRK4 gene polymorphisms with essential hypertension in obese ethnics Uygur from southern Xinjiang / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of polymorphisms of G protein-coupled inwardly-rectifying potassium channels 4 (GIRK4) gene with essential hypertension in ethnic Uygurs from southern Xinjiang.</p><p><b>METHODS</b>A total of 1194 (461 males and 733 females) Uygur residents aged 30 to 70 and with a body mass index (BMI) over 18.5 kg/m(2) were selected from Hetian region. All of the subjects have received questionnaire survey, physical examination, biochemical analysis and blood pressure measurement. They were divided into hypertensive group and normotensive group. Genotyping by the TaqMan polymerase chain reaction method was performed for 4 common single nucleotide polymorphisms (rs4937391, rs2604204, rs6590357 and rs1122149), and a case-control study was carried out.</p><p><b>RESULTS</b>Genotype distributions of rs4937391, rs2604204, rs6590357 and rs1122149 in both groups were in Hardy-Weinberg equilibrium (P> 0.05). The average systolic blood pressure of CC genotype of rs11221497 single nucleotide polymorphism (SNP)[(132.69± 26.9) mmHg)] was higher than the CG genotype [(127.4± 22.7) mmHg] and GG genotype [(121.1± 26.3) mmHg]. There has a significantly difference in average systolic and diastolic blood pressures between CC and GG genotypes (P< 0.05). A case-control association analysis revealed that the rs11221497 SNP was in association with essential hypertension with the dominant model [P< 0.05, OR= 0.67 (0.49-0.93)]. Haplotype analysis indicated that H6(C-G-C-G) was significantly more common in normotensive group than hypertensive group (P= 0.001).</p><p><b>CONCLUSION</b>The rs11221497 SNP of the GIRK4 gene is associated with essential hypertension in ethnic Uygur population in Xinjiang.</p>

Association of SOCS3 gene polymorphisms with insulin resistance in Xinjiang Uygur population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To assess the association of suppressor of cytokine signaling 3 (SOCS3) gene polymorphisms with insulin resistance (IR) in Xinjiang Uygur population.</p><p><b>METHODS</b>A cross-sectional study on metabolic diseases, e.g., obesity, was conducted on ethnic Uygurs in Hetian, Xinjiang of China. A total of 1292 Uygur individuals were enrolled. The sample size for IR subjects [homeostasis model assessment for insulin resistance (HOMA-IR) ≥ 2.96] was 323, whereas that for non-IR controls was 969 (HOMA-IR < 2.96). Representative variations were selected from database and genotyped with TaqMan polymerase chain reaction method. For the relatively isolated population from a homogeneous environment, a case-control study was conducted to assess the association between variations of SOCS3 gene and IR.</p><p><b>RESULTS</b>A significant difference in genotype distribution of rs4969168 was detected between IR and control groups in males (chi square =7.216, P=0.027). Although the IR-related quantitative phenotypes did not significantly differ between individuals with GG, AG and AA genotypes of rs4969168 in the overall, male and female population (P > 0.05), the mean of body mass index and the median of fasting insulin increased in individuals with GG, AG, AA genotypes in males. Haplotype 2 (rs12953258C-rs4969168A-rs9914220C) was significantly associated with a higher prevalence of IR in males (P=0.023). Logistic regression analysis indicated that AG genotype of rs4969168 variation is a protective factor for IR in males (OR=1.772, 95% confidence interval: 1.081-2.906, P=0.023).</p><p><b>CONCLUSION</b>Our study suggested that the rs4969168 polymorphism of SOCS3 gene is associated with IR in ethnic Uygur population from Xinjiang, China.</p>

Relationship between SOCS-3 gene polymorphism and abnormal glucose metabolism in the Xinjiang Uygur population / 中华内分泌代谢杂志

Objective To investigate the relationship between SOCS-3 gene polymorphism and abnormal glucose metabolism in Xinjiang Uygur population.Methods According to different plasma glucose levels,1232individuals in Xinjiang Hetian area were divided into 3 groups,451 patients with pre-diabetes,252 patients with type 2 diabetes mellitus,and 529 healthy people as normal controls.Based on HapMap,the polymorphisms rs12953258,rs4969168,rs9914220 were selected as haplotypes,tagging SNP (htSNP) sufficiently covering the genetic variation of the whole gene.Associations of rs12953258,rs4969168,and rs9914220 within SOCS-3 with abnormal glucose metabolism in three groups of Xinjiang Uygur population were examined ; The genotype and allele frequencies and relative clinic data were compared among groups.Results The type 2 diabetes mellitus study with 451 individuals showed that the homozygosity for the C allele of rs12953258 polymorphism of SOCS-3 was associated with increased diabetes risk(OR=1.756,95％ CI 1.168-2.640).In addition,association between rs4969168 or rs9914220 and abnormal glucose metabolism in the Xinjiang Uygur population was not found.Age,total cholesterol,and body mass index were risk factors of diabetes mellitus,total cholesterol and low high density lipoprotein-cholesterol were risk factors of pre-diabetes in Uygur people.Conclusions The C allele of rs12953258 polymorphism of SOCS-3 gene may be an independent risk factor for abnormal glucose metabolism in Xinjiang Uygur population.

Association between the regulator of G-protein signaling 2 gene 1891-1892del TC polymorphism with hypertension in Xinjiang Kazakh population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of the 1891-1892del TC polymorphism of regulator of G-protein signalling 2 (RGS2) gene with hypertension in Xinjiang Kazakh population.</p><p><b>METHODS</b>The case-control study was performed in 444 cases and 489 controls. The genotypes of the individuals in the 1891-1892del TC locus were identified by TaqMan method.</p><p><b>RESULTS</b>A significant association was observed between the1891-1892TC insertion/deletion polymorphism with hypertension in men (OR=1.698, P=0.03) and in the total population (OR=1.32, P=0.044). The mean systolic blood pressure and serum uric acid levels of the ID+DD carriers were significantly higher than that of the II carriers (adjusted, P=0.04 and P<0.01).</p><p><b>CONCLUSION</b>The results suggest that the D allele of the 1891-1892TC insertion/deletion locus of the RGS2 gene might be an independent risk factor for hypertension in Xinjiang Kazakhs; and the polymorphism may have some influence on serum uric acid level in this population.</p>

Determination of Paeoniflorin in Yifu Capsules by HPLC / 中成药

Objective: To establish a method of HPLC for determination of paeoniflorin in Yifu Capsules. Methods: HPLC analysis was carried out on ALLIMA C 18 column with acetonitrile water(15:85) as a mobile phase and detection wavelength at 230nm. Results: The linear range was 0.0798～0.7980?g (?=0.9998,n=6), and the average recovery was 99.00%(RSD=2.17%,n=5). Conclusion: The method is simple, accurate,highly sensitive and reproducible. It can be used for determination of paeoniflorin in Yifu Capsules.