1.Effects of the serum in shock rats and shenfu injection on the endothelial cell protein C receptor
Yueyue HUANG ; Jingye PAN ; Guanyang LIN ; Jinbo ZHANG ; Xiaole ZHANG ; Haifeng JIN ; Zhiqiang LI ; Jie CHEN ; Jihong MA
Chinese Journal of Emergency Medicine 2011;20(2):168-172
Objective To investigate the effects of the serum from rats with hemorrhagic shock and Shenfu injection, on the expression of endothelial cell protein C receptor (EPCR) in human umbilical vein endothelial cells (HUVECs) cultured with rat serum. Method The soluble endothelial protein C receptor (sEPCR) in supernatant, the expression of EPCR mRNA and protein level of EPCR in HUVECs were detected by using enzyme linked immunosorbent assay ( ELISA), reverse transcription polymerase chain reaction (RT-PCR), and western bloting (WB) in normal control group, hemorrhagic shock serum (3 h, 12 h, 24h, 72 h) group, and Shenfu-treated (3 h, 12 h, 24 h, 72 h) group, respectively. Results The mean levels of sEPCR and the expression of EPCR mRNA were significantly higher in hemorrhagic shock serum (12 h, 24 h) group, and Shenfu -treated(24 h)group than those in normal control group (all P <0.01 ),the mean levels of sEPCR and the expressions of EPCR mRNA were significantly higher in Shenfu-treated ( 12 h) group than those in normal control group ( all P <0. 05 ), while the levels of protein were lower in hemorrhagic shock serum ( 12 h, 24 h) group and in Shenfu-treated(24 h)group than those in normal control group ( both P <0.01 ), and the level of EPCR protein was lower in Shenfu-treated( 12 h) group than that in normal control group ( P < 0. 05) . The mean levels of sEPCR and the expressions of EPCR mRNA were significantly lower in Shenfu-treated ( 12 h, 24 h) group than those in hemorrhagic shock serum ( 12 h,24 h) group (all P <0.05), while the levels of EPCR protein were higher in Shenfu-treated ( 12 h, 24 h)group than those in hemorrhagic shock serum ( 12 h, 24 h) group ( P < 0.05 ). Conclusions These data suggest that Shenfu injectio could affect the expression of EPCR mRNA and the level of EPCR protein, thereby it might be effective in prevention of development of hemorrhagic shock.
2.lncRNA XIST regulates the malignant biological behaviors of colorectal cancer HCT-8 cells through miR-32-5p/EZH2 molecular axis
WU Ruiping ; CHEN Guanyang ; CHEN Zhikang
Chinese Journal of Cancer Biotherapy 2019;26(12):1363-1370
Objective: To explore the mechanism of lncRNA XIST (XIST) regulating the biological behaviors of colorectal cancer HCT-8 cells via miR-32-5p/EZH2 (enhancer of Zeste homolog 2) axis. Methods:Atotal of 28 pairs of cancer tissues and corresponding para-cancerous tissues form colorectal cancer patients with complete clinical data were collected from the Colorectal and Anal Surgery, Xiangya Hospital of Central South University during July 2014 and August 2018. The expression levels of lncRNA XIST and miR-325p in colorectal cancer tissues and cell lines were detected by qPCR. The targeted relationship between lncRNA XIST, miR-32-5p and EZH2 was verified by dual luciferase reporter gene, and the expression level of EZH2 was further detected by WB. The proliferation, migration and apoptosis of HCT-8 cells were detected by CCK-8, Transwell and flow cytometry with Annexin V-FITC/PI staining, respectively. Results: lncRNAXIST was highly expressed in colorectal cancer tissues and cell lines with the highest expression in HCT-8 cells (P<0.05 or P<0.01). Dual luciferase reporter gene assay validated that lncRNA XIST negatively regulated miR-32-5p (P<0.05), and EZH2 was a target gene of miR-32-5p. Knockdown of lncRNAXIST inhibited proliferation and migration and induced apoptosis of HCT-8 cells (P<0.05 or P<0.01). Further experiments demonstrated that knockdown of lncRNA XIST up-regulated the expression of miR-32-5p and further down-regulated the expression level of EZH2, thereby inhibiting the proliferation and migration of HCT-8 cells and inducing apoptosis. Conclusion: lncRNAXIST promotes proliferation, migration and inhibits apoptosis of HCT-8 cells via miR-325p/EZH2 axis.
3.Comparison of various prediction models in the effect of laparoscopic sleeve gastrectomy on type 2 diabetes mellitus in the Chinese population 5 years after surgery
Chengyuan YU ; Liang WANG ; Guangzhong XU ; Guanyang CHEN ; Qing SANG ; Qiqige WUYUN ; Zheng WANG ; Chenxu TIAN ; Nengwei ZHANG
Chinese Medical Journal 2024;137(3):320-328
Background::The effect of bariatric surgery on type 2 diabetes mellitus (T2DM) control can be assessed based on predictive models of T2DM remission. Various models have been externally verified internationally. However, long-term validated results after laparoscopic sleeve gastrectomy (LSG) surgery are lacking. The best model for the Chinese population is also unknown.Methods::We retrospectively analyzed Chinese population data 5 years after LSG at Beijing Shijitan Hospital in China between March 2009 and December 2016. The independent t-test, Mann–Whitney U test, and chi-squared test were used to compare characteristics between T2DM remission and non-remission groups. We evaluated the predictive efficacy of each model for longterm T2DM remission after LSG by calculating the area under the curve (AUC), sensitivity, specificity, Youden index, positive predictive value (PPV), negative predictive value (NPV), and predicted-to-observed ratio, and performed calibration using Hosmer–Lemeshow test for 11 prediction models. Results::We enrolled 108 patients, including 44 (40.7%) men, with a mean age of 35.5 years. The mean body mass index was 40.3 ± 9.1 kg/m 2, the percentage of excess weight loss (%EWL) was (75.9 ± 30.4)%, and the percentage of total weight loss (% TWL) was (29.1 ± 10.6)%. The mean glycated hemoglobin A1c (HbA1c) level was (7.3 ± 1.8)% preoperatively and decreased to (5.9 ± 1.0)% 5 years after LSG. The 5-year postoperative complete and partial remission rates of T2DM were 50.9% [55/108] and 27.8% [30/108], respectively. Six models, i.e., "ABCD", individualized metabolic surgery (IMS), advanced-DiaRem, DiaBetter, Dixon et al’s regression model, and Panunzi et al’s regression model, showed a good discrimination ability (all AUC >0.8). The "ABCD" (sensitivity, 74%; specificity, 80%; AUC, 0.82 [95% confidence interval [CI]: 0.74–0.89]), IMS (sensitivity, 78%; specificity, 84%; AUC, 0.82 [95% CI: 0.73–0.89]), and Panunzi et al’s regression models (sensitivity, 78%; specificity, 91%; AUC, 0.86 [95% CI: 0.78–0.92]) showed good discernibility. In the Hosmer–Lemeshow goodness-of-fit test, except for DiaRem ( P <0.01), DiaBetter ( P <0.01), Hayes et al ( P = 0.03), Park et al ( P = 0.02), and Ramos-Levi et al’s ( P <0.01) models, all models had a satifactory fit results ( P >0.05). The P values of calibration results of the "ABCD" and IMS were 0.07 and 0.14, respectively. The predicted-to-observed ratios of the "ABCD" and IMS were 0.87 and 0.89, respectively. Conclusion::The prediction model IMS was recommended for clinical use because of excellent predictive performance, good statistical test results, and simple and practical design features.