2.Condition of human Th17 ceII differentiation induced in vitro
Chen WANG ; Wei GUO ; Xin WANG ; Dongmei YU ; Yuheng WANG ; Wenbing YAO
Journal of China Pharmaceutical University 2016;(1):106-111
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.
3.Effects of sevoflurane postconditioning on myocardial ischemia-reperfusion injury in patients undergoing coronary artery bypass grafting with cardiopulmonary bypass
Chunxia SHI ; Lihuan LI ; Yuntai YAO ; Xin WANG ; Min SONG ; Cuntao YU ; Yingmao RUAN
Chinese Journal of Anesthesiology 2010;30(12):1431-1434
Objective To investigate the effects of sevoflurane postconditioning on myocardial ischemiareperfusion(I/R)injury in patients undergoing coronary artery bypass grafting(CABG)with cardiopulmonary bypass(CPB).Methods Forty NYHA Ⅰ -Ⅲ patients of both sexes,aged 55-64 yr,with BMI < 30 kg/m2,scheduled for CABG under CPB,were randomly divided into 2 groups(n = 20): control group(group C)and sevoflurane postconditioning group(group S).Anesthesia was induced with midazolam and/or etomidate,fentanyl and rocuronium.Patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with iv infusion of propefol and intermittent iv injection of fentanyl and pipecuronium.In group S,2% sevoflurane was inhaled continuously for 15 min immediately after aortic unclamping.After anesthesia induction,before CPB,10 min after the end of CPB,at the end of operation,and 6 and 24 h after operation,MAP,HR,CVP,mean pulmonary arterial pressure,pulmonary arterial wedge pressure,CO and S(v)O2 were recorded,and CI,SVI,systemic vascular resistance index and pulmonary vascular resistance index were calculated.Blood samples were taken from central vein before aortic clamping,at 6 h of reperfusion and 24 h after operation for determination of plasma creatine kinase(CK),creatine kinase isoenzyme(CK- M B)and lactate dehydrogenase(LDH)activities and tropenin I(TnI)concentrations.Myocardial tissues were obtained from right auricle before aortic clamping and at the end of CPB for observation of the ultrastructure and the severity of myocardial injury was assessed.Results There was no significant difference in hemodynamics and parameters of cardiac function between the two groups(P > 0.05).Compared with group C,plasma CK-MB and LDH activities at 6 h of reperfusion and plasma CK activity and TnI concentrations at 24 h after operation were significantly decreased and the myocardial injury was significantly reduced after the end of CPB in group S(P < 0.05).Conclusion Sevoflurane postconditioning can protect myocardium against I/R injury induced by CPB in patients undergoing CABG.
4.An analysis on the prevalence and risk factors of post-transplant diabetes mellitus after renal transplantation
Minling CHEN ; Yao ZHANG ; Mingxiang YU ; Ming XU ; Jian GAO ; Xin GAO
Chinese Journal of Endocrinology and Metabolism 2013;29(9):750-755
Objective To explore the incidence of post-transplant diabetes mellitus (PTDM) by means of fasting plasma glucose (FPG) and other associated risk factors in patients surviving for more than 1 year after renal transplantation.Methods A total of 428 non-diabetic patients,who underwent kidney transplantation from 1 January,1993 to 31 December,2008,were followed up in order to ascertain the prevalence of PTDM after transplantation and other associated risk factors by means of FPG.Results Of the 428 patients,87 developed PTDM (20.3%) within a mean follow-up of (5.65 ± 3.68) years after renal transplantation.The onset of PTDM occurred in 57 patients (65.5% of total PTDM) primarily within the first year after transplantation.Univariate analysis showed that older age,body mass index (BMI),smoking history,family history of diabetes mellitus,deceased donor transplantation,hepatitis C virus infection,cytomegalovirus infection,FPG before transplantation as well as 1 week after transplantation,total cholesterol and triglyceride before transplantation,switching from cyclosporine to tacrolimus(FK506),and peak plasma concentration of cyclosporine in the first 6 and 12 months were associated with the onset of PTDM.The prevalence of PTDM was markedly elevated in the group who has cyclosporine converted to FK506 (P<0.05),but not in the group with cyclosporine converted to rapamycin.By multivariate analysis,FPG before transplantation,age,BMI,and deceased donor transplantation were independently associated with the onset of PTDM.Conclusions There is high incidence of PTDM in patients following renal transplantation; and early diagnosis,treatment as well as prevention are mandatory.
5.Effects of delivery mode on postpartum fecal incontinence and urinary incontinence in Chinese women
Hongxia ZHANG ; Xin YANG ; Hairong YAO ; Rui WANG ; Xiaohua LI ; Haiyang YU ; Huixia YANG ; Yue DONG
Chinese Journal of Perinatal Medicine 2011;14(10):598-602
Objective To investigate the prevalence and associated factors of fecal incontinence (FI) and urinary incontinence (UI) in postpartum Chinese women.Methods Questionnaires about FI and UI symptoms were completed via telephone interviews conducted within six months after delivery.Multi-variant Logistic analysis was applied for relation between delivery mode and FI or UI.Results (1) Two thousand and twelve postpartum women were admitted into this study,among which 14 (0.7%) had FI within 6 months after delivery.Logistic regression analysis showed that FI was significantly associated with forceps delivery (OR =20.09,95 % CI:3.64-110.90,P =0.000),and mediolateral episiotomy (OR=6.11,95% CI:1.29-28.80,P=0.024).(2) Among the 2012 women,the prevalence of UI,stress urinary incontinence (SUI),urgent urinary incontinence (UUI)and mixed urinary incontinence (MUD was 10.04% (n=202),8.15% (n=164),0.94% (n=19)and 0.94 % (n =19),respectively.Logistic regression analysis found that SUI prevalence was related to maternal age (OR =1.07,95% CI:1.04-1.11,P =0.000),maternal weight before delivery (OR=1.04,95% CI:1.02-1.06,P=0.001),neonatal head circumference (OR=1.20,95% CI:1.05-1.39,P =0.010),mediolateral episiotomy (OR =4.96,95 % CI:3.05-8.07,P =0.0005 ),spontaneous vaginal delivery (OR=5.22,95% CI:2.53-10.76,P=0.000) and forceps delivery (OR=9.20,95% CI:4.07-20.79,P=0.000).UUI was related to maternal weight before delivery (OR=1.51,95% CI:1.12-2.05,P=0.008).MUI was related to maternal weight before delivery (OR=1.06,95% CI:1.00-1.11,P=0.049),duration of second stage of labor (OR=1.01,95% CI:1.00-1.03,P =0.010),mediolateral episiotomy (OR =7.76,95% CI:1.42-42.52,P=0.017) and forceps delivery (OR=15.21,95% CI:1.61-143.44,P=0.018).(3) The prevalence of SUI was higher at 4 days and 42 days after delivery (7.95% and 9.10%).Conclusions (1) F1 and UI prevalence is lower in this study than in other reports.(2) Vaginal delivery is a risk factor for women's FI and UI,especially forceps delivery and mediolateral episiotomy.(3) Maternal age,pre-delivery weight,newborn head circumference,spontaneous vaginal delivery,forceps delivery and mediolateral episiotomy might increase the risk of UI.
6.Investigation on bibliotherapy in nursing care of perioperative children
Hanping GUO ; Chunhua YU ; Hua ZHANG ; Tianhong LI ; Cuiping XIAO ; Xin CHEN ; Wenyan YAO
Chinese Journal of Practical Nursing 2010;26(26):6-9
Objective To investigate the effect of bibliotherapy in soothing the postoperative pain in children and relieving the perioperative anxiety of children and their care givers. Methods Hospitalized children and their care givers from August 2007 to March 2009 were studied. 153 cases from August 2007 to May 2008 were assigned to the control group and the other 153 cases to the intervention group. Routine surgical nursing were applicated in the control group by introduction of perioperative nursing procedures.Bibliotherapy were applicated in the intervention group on the basis of the control group-using "bibliotherapy materials for hospitalized children" which was designed by ourselves and correspond with the theory of bibliotherapy to interfere in the 153 cases in the intervention group. The variance of preoperative anxiety of care givers and perceptions of postoperative pain of children between the two groups were compared with scales of mYPAS, STAI and FLACC and Wong- Baker Facial Scale. Results The scores of mYPAS in children of the intervention group and the control group was (35.875+4.441)and(46.796+8.606 )respectively and the variance was significant. The scores of STAI in care givers of the two groups was(38.125+4.371 )and (49.901 +7.420) respectively and revealed significant variance. The scores of Wong-Baker and FLACC in children of both groups 1 hour after operation were compared and revealed no statistical significance. The scores of Wong-Baker and FLACC in children at 6 hours and 24 hours postoperative were compared subjectively and objectively and revealed statistic significance. Conclusions Bibliotherapy can ameliorate the anxiety level of both children and their care givers, relieve the perception of postoperative pain in children and improve their comforts. Bibliotherapy thus conduces to the recovery of postoperative children.
7.Protective effect of dl-praeruptorin A on focal cerebral ischemia in mice
Wushuang YANG ; Bogang TENG ; Lichao YANG ; Yu ZHOU ; Yao WANG ; Xin JIN
Chinese Journal of Biochemical Pharmaceutics 2010;31(2):118-121
purpose To investigate the protective effect and character of dl-praeruptorin A(Pd-Ia)on focal cerebral ischemia in mice.Methods Transient focal cerebral ischemia in mice WaS induced by middle cerebral artery occlusion for 1.5 h.Pd-Ia was administered intraperitoneally either with multiple doses(1,5 and 10ms/ks)at 0.5 h before ischemia or single dose(5 ms/kg)at 0.5 h and 1 h before ischemic,the same time of ischemia,the same time of reperfusion,or 0.5 h and 1 h after reperfusion respectively.Neurological deficit score,infarct volume,brain edema,the activities of SOD and the contents of MDA were determined.Results Pretreatment with multiple doses(5 and 10 ms/ks)of Pd-Ia at 0.5 h before ischemia or single dose(5 mg/kg)of Pd-Ia at 0.5 h before ischemia,at the same time of ischemic,at the same time of reperfusion and 0.5 h after reperfusion significantly attenuated neurological deficit score,decreased infarct volume and alleviated brain edema,and the treatment at the time of reperfusion had the most marked effect.Pd-Ia(5 or 10 ms/ks)can significantly enhance the activities of SOD and lower the contents MDA.Conclusion dl-praeruptorin A has a neuroprotective effect on the injury in the acute phase of transient focal cerebral ischemia in mice,with optimal doses of 5 ms/ks and the optimal therapeutic time point of the same time of reperfusion.
8.Comparing effects of U50488H, prazosin and/or propranolol on cardiac hypertrophy induced by NE in rat.
Gui-jun WANG ; Yu-sheng YAO ; Hong-xin WANG
Chinese Journal of Applied Physiology 2010;26(1):82-85
OBJECTIVETo demonstrate the inhibitory effect of kappa-opioid receptor activation by U50488H on hypertrophy induced by NE in cultured neonatal rat cardiac myocytes and compare its effect with that of prazosin and propranolol.
METHODSThe cellular proliferation was determined with crystal violet staining. The protein content was assayed with Lowry's method. The cardiomyocytes volumes were measured by computer photograph analysis system. The protein synthesis was assayed with [3H]-lencine incorporation method.
RESULTS(1) NE significantly induced the increase of protein content, [3H]-leucine incorporation and cell size without a concomitant increase in cell number in low serum medium. OThese responses were partially suppressed by prazosin or propranolol alone and completely abolished by both in combination. U50488H significantly inhibited the NE-induced increase of protein content, [3H]-leucine incorporation and cell size. The inhibitory effects of U50488H on NE-induced cardiac hypertrophy were greater than either prazosin or propranolol, but comparable to combination of both.
CONCLUSIONNE, acting via both alpha1- and beta-adrenergic pathway, stimulates myocyte hypertrophy. Stimulating kappa-opioid receptor significantly inhibits NE-induced cardiac hypertrophy, which may be related with alpha1- and beta1-adrenergic pathway.
3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer ; pharmacology ; Adrenergic alpha-1 Receptor Antagonists ; pharmacology ; Adrenergic beta-Antagonists ; pharmacology ; Animals ; Animals, Newborn ; Cardiomegaly ; chemically induced ; pathology ; prevention & control ; Cell Enlargement ; drug effects ; Cells, Cultured ; Female ; Male ; Myocytes, Cardiac ; cytology ; Norepinephrine ; Prazosin ; pharmacology ; Propranolol ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, kappa ; agonists