In this study,a series of related indicators were investigated via flow cytometry,enzyme-linked immu-no sorbent assay (ELISA)and quantitative real time PCR (Q-PCR)technology to assess the in vitro differentia-tion of human Th17 cells.Human peripheral blood mononuclear cells (PBMCs)were purified from fresh human blood using gradient centrifugation and then the Th17 cells were induced with different cytokines (IL-1β,IL-6, TGF-βand IL-23)at different induction time (1,2,3,4 d)to compare the effects on Th17 cell differentiation un-der these conditions.The experiment data showed that IL-1β,IL-6,TGF-βor IL-23 alone play a promotion role in the Th17 differentiation and combination of IL-1β,IL-6,TGF-βand IL-23 could induce efficient human Th17 cell differentiation in vitro to achieve the best.Further optimization of the induction time found that the Th17 cell dif-ferentiation efficiency gradually increased with the extension of the time;howerver,when culturing for 3 d,it reached the peak number and then decreased in regardless of the time increasion.Finally the optimal condition of in vitro polarization of human Th17 cells was established,and the purified PBMCs were cultured with anti-CD3 and anti-CD28 as the basal conditions,and co-culturing with IL-1β,IL-6,TGF-βand IL-23 for 3 d to effectively induce the differentiation of Th17 cells.The inducing efficiency is significantly higher than in normal control.At the optimal condition,we observed the Th17 cell differentiation frequency (CD4 +IL17A +)was increased to nearly 10% through flow cytometry analysis and the secretion level of IL-17A in cell supernatants was also detec-ted to reach 3 ng/mL using ELISA methods.In addition,gene expression of IL-17A were determined by quantitativereal-time PCR using pre-designed primers by the comparative method of relative quantitation (ΔΔCt)and β-actin gene was used as an internal control for sample normalization.The results showed that the expression of IL-17A mRNA could be increased about 15 times with IL-1β,IL-6,TGF-βand IL-23 co-culturing for 3 d.The protocolof efficient human Th17 cell differentiation we presented in this paper is simple,rapid and easy to be repeated.This study provides an effective detection platform for the research of Th17 cell function and development ofrelated drugs targeting Th17 cells for autoimmune disease treatment.

Objective To study the biological effects of (Janus Kinase-Signal transducers and activators of transcription,AK-STATs)path-way in hair cell apoptosis by analyzing the expression level of signal transducers and activators of transcription (STATS) of apoptotic HEI-OC1 cells line induced by oxidative stress injury.Methods Using different poisoning concentrations (0,20,40,60μM) of(Tert-Butyl hydroperoxide,T-BHP) built the apoptotic HEI-OC1 cells model while 0 μM was used as the control group.The apoptosis level at different poisoning concentration groups was detected by Annexin V-FITC/PI and the expression level of STATs mRNA by real-time quantitative PCR detecting system.Results The apoptosis rates in different contamination groups(0,20,40,60 μM) were 3.9％,7.4％,32.0％,and 91.2％,respectively.Compared with control group,STAT1 mRNA,STAT3 mRNA,STATS mRNA,and STAT6 mRNA expression levels in different poisoning concentration groups declined significantly (F=5 534.302,P＜0.01;F=146.038,P＜0.01;F=685.929,P＜0.01;F=516.11,P＜ 0.01).No statistically significant differences were found among the poisoning concentration groups (P＞0.05).Compared with the control group,the STAT2 mRNA expression levels changed significantly (F=1 259.148,P＜ 0.01).The STAT2 mRNA expression level in 20 μM contamination group decreased while 40,60 μM contamination group increased significantly (P＜0.01).No expression of STAT4 in HEI-OC1 cell was noted.Conclusion The JAK-STAT2 signal pathway has the biological effects on leading oxidative stress injury apoptosis and JAK-STAT1/STAT3/STAT5/STAT6 has the biological effect of inhibit oxidative stress injury apoptosis.

Objective To evaluate the effectiveness of perioperative fasting abbreviation in traumatic patients undergoing selective surgeries.Methods The traumatic patients undergoing selective surgeries from November 2016 to January 2017 at our department were selected for this prospective cohort study.They were divided into an intervention group (69 patients) and a control group (121 patients) according to the wards where they stayed.The intervention group was fasted for solids 6 hours prior to surgery and received oral solution with maltodextrin 2 hours prior to surgery.After surgery,they were allowed to drink liquids as soon as they were awakened.Normal food was allowed 2 hours later.The control group was fasted for either liquids or solids the night before surgery.After surgery,the patients who had received brachial plexus block only were allowed liquids with no limitation while the other patients were allowed liquids 6 hours after surgery and then were free for solids and liquids if no discomfort was observed.The time periods for preoperative liquids and solids fasting and for postoperative intake of liquids and solids were recorded and compared between the 2 groups.The perioperative well-beings (including anxiety,thirst,hunger,nausea,fatigue,dizziness,sweating and stomach discomfort) and serum glucose levels were compared between the 2 groups.Adverse reactions were observed.Results The preoperative fasting time for liquids for the intervention group (4.5 ± 2.9 hours) was significantly shorter than that for the control group (14.3 ±3.9 hours) (P ＜ 0.05).The preoperative fasting time for solids for the intervention group (17.6 ± 3.0 hours) were significantly longer than that for the control group (16.1 ±3.8 hours) (P ＜ 0.05).The postoperative fasting time periods for both liquids [1 (0,3) h] and solids [2 (1,4) h] for the intervention group were significantly shorter than those for the control group [6(6,6) h] hours and [6(6,6) h] (P ＜ 0.05).Compared with the control group,the perioperative anxiety,thirst,hunger,nausea,fatigue,dizziness and stomach discomfort were significantly improved in the intervention group (P ＜ 0.05).The average serum glucose level was similar in both groups upon admission (P ＜ 0.05);it was significantly higher in the intervention group immediately before surgery (P ＜ 0.05) but was gradually decreased after surgery until there was no significant difference between the 2 groups (P ＞ 0.05).No major adverse reaction was observed in either group.Conclusion The protocol of perioperative fasting abbreviation may be safe and feasible in traumatic patients for selective surgeries,showing benefits of decreased anxiety,thirst,hunger,nausea,fatigue,dizziness and stomach discomfort.