1.The influence of swallowing autoantigen on differentiation,development and function of the bone marrow-derived DC in vitro
Jun XIE ; Youkun LIN ; Hong LUO
Chinese Journal of Immunology 2000;0(09):-
Objective:To observe the changes in differentiation,development and function of DCs after mouse bone marrow-derived dendritic cells devouring autoantigen in vitro,to study its impact on the immune status.Methods:Mouse bone marrow cells were cultured in GM-CSF+IL-4 to generate DCs,and the purity of DCs was detected by fluorescence microscope and flow cytometry.It was identified whether DCs could swallow autoantigen by immunohistochemistry and transmission electron microscope;and the changes in CD86,MHCⅡ on DCs after swallowing autoantigen were observed by flow cytometry.The changes in proliferation after the mouse spleen lymphocytes were cultured with the same genetic background of DCs were measured by MTT.The amounts of IL-4,IFN-? secreted by the mouse spleen lymphocytes after stimulation with DCs were measured by ELISA.Results:The purity of DCs was 90.6%.DCs could swallow autoantigen,which induced differentiation dysplasia of the DCs in vitro.Swallowing autoantigen had negative effect on the maturation as well as the potentiation to stimulate proliferation of the mouse spleen lymphocytes with the same genetic background.Secretion of IFN-? by stimulated lymphocytes decreased,but secretion of IL-4 promoted.Conclusion:The DCs that swallow autoantigen could inhibit imDCs maturation,but promote the immune tolerance and humoral immunity.
4.Expression of PTTG and its relationship with bFGF and MVD in esophageal carcinoma
Jialong GUO ; Chengyi LIN ; Shunqing ZUO ; Jun ZHANG ; Bingsheng XIE
Journal of Chinese Physician 2008;10(9):1162-1165
Objective To study the expressions of PTTG and bFGF proteins and their relationship with microvessels density(MVD)in esophageal carcinoma.Methods Immunohistochemical SP method was used to detect the expression of PTTG and bFGF proteins in 48 esophageal carcinoma tissues and the same para-cancerous tissues.MVD was evaluated by immunohistochemieal staining with antibody CD34.Results The positive rate of PTTG and bFGF was 68.8%(33/48)and 70.8%(34/48)respectively.Rate of PTTG protein expression in esophageal carcinoma tissues was significantly higher than that in para-cancerous tissues(8.3%and 12.5%,P<0.05).The positive rate 0f PTTG,bFGF and MVD was correlated with lymph node metastasis and TNM stage.There was no relationship with age,sex,tumor size MVD(P<0.05).Conclusion PTTG and bFGF are over-expressed in esophageal carcinoma.Increased PTTG may play an important role in carcinogenesis and development of esophageal carcinoma by promoting the expression of bFGF protein which may induce an angiogenesis.
5.Effects of transforming growth factor-β2 on human Tenon fibroblasts transformation and scarring after glaucoma filtration surgery
Xiao-yan, ZHU ; Lei, LI ; Guang-jun, XIAN ; Hai-jun, LI ; Yan, TAN ; Lin, XIE
Chinese Journal of Experimental Ophthalmology 2013;(3):215-219
Background Research showed that transforming growth factor-β2 (TGF-β2) promotes scar formation.But its mechanism in scarring after glaucoma filtration surgery is worthy of studying.Objective This study was to investigate the effect of TGF-β2 on myofibroblast transition of human Tenon fibroblasts (HTFs) and scarring after glaucoma filtration surgery.Methods Tenon capsular tissue was obtained from 3 patients with strabismus during the surgery and was incubated in DMEM with 10% fetal bovine serum (FBS).The cells were collected and passaged in the free-serum medium for 24 hours,and then 1,2,5,10,20 μg/L TGF-β2 was added into the medium respectively,to induce the transformation of HTFs,and 2 μg/L or 5 μg/L TGF-β2 was used to treat the HTFs for 6,24,48 and 72 hours.The control group was not treated with TGF-β2.The expressions of α-smooth muscle actin (α-SMA) and phosphorylation of the signaling proteins (pSmad2) in HTFs were detected by Western blot assay.The expressions of α-SMA and F-actin were located by cell immunofluorescine technique under the confocal immunofluorescence microscopy.Cell contractility was determined by collagen gel contraction assays.This study was approved by Ethic Committee of Institute of Surgery Research of Daping Hospital,and informed consent was obtained from each patient or custodian initial of the study.Results The expression of α-SMA protein in the HTFs was increased significantly after the treatment of TGF-β2 in comparison with the control group and reached a peak at 24-48 hours.The α-SMA expression was gradually weakened in the 10 μg/L TGF-β2 groups.Little of α-SMA and F-actin were expressed in the control group.However,strong staining for α-SMA and F-actin were observed in the 1,2 and 5 μg/L TGF-β2 groups and then the staining weakened at the concentration of 10 μg/L.In addition,pSmad2 showed a stronger expression in the 2 μg/L TGF-β2 group than that in the PBS group and FBS group,with the strongest expression in 30 minutes through 2 hours.The untreated gel contracted (78.00±3.13)% from its initial size,and contraction in the 1,2,5,10 μg/L TGF-β2 group were (63.88±1.78)%,(20.69±0.65)%,(19.49-±0.54)%,(16.24±0.84) %,respectively,TGF-β2 increased HTFs contraction significantly (Fgroup =859.400,P =0.000).Conclusions TGF-β2 can induce transdifferentiation of Tenon fibroblast into myofibroblast and increase cell contractility,with a concentration-dependent and time-dependent pattern to an extent.It may be the mechanism of scar formation after glaucoma filter surgery.
6.Indomethacin for prevention of post-ERCP pancreatitis and hyperamylasemia
Tao LIN ; Yi ZHOU ; Jun WANG ; Ru YAO ; Li JIANG ; Rutang FANG ; Suli WANG ; Yan XIE
Chinese Journal of Digestive Endoscopy 2012;29(4):185-187
Objective To investigate the preventive effect of Indomethacin for post-ERCP pancreatitis and hyperamylasemia.Methods A total of 600 patients,who were undergoing ERCP,were randomly divided into 3 groups to receive anal Indomethacin (n=200),intravenous octreotide (n=200) or no special medication (n=200) before ERCP.The level of serum amylase before and 24h after ERCP were measured,and the rate of acute pancreatitis and hyperamylasemia after ERCP were assessed.Results Serum amylase levels before ERCP of all groups were normal.The mean serum amylase level of Indomethacin group (101.3±77.7 U/L) after ERCP was significantly lower than those of octreotide group ( 176.6±138.3 U/L,P =0.040 ]and control group (227.2±264.9 U/L,P=0.048),while there was no difference between octreotide group and control group ( P>0.05 ).The incidence of post-ERCP pancreatitis in Indomethacin group (2.5%) was significantly lower than that of control group (9.5%,P=0.003),while there was no difference between octreotide group (4.5%) and control group ( P=0.05 ).The incidence of hyperamylasemia after ERCP in Indomethacin group (5.5%) was significantly lower than that of control group ( 13.5%,P=0.006 ),while there was no difference between octreotide group (10.0%) and control group ( P>0.05 ).Conctusion Anal administration of Indomethacin before ERCP can effectively reduce the incidence of acute pancreatitis and hyperamylasemia after ERCP.
7.Correlation of the right ventricular systolic function indexes evaluated by two-and three-dimensional echocardiography with physical development in normal children
Jun GAO ; Mingxing XIE ; Qing Lü ; Lingyun FANG ; Lin HE ; Li ZHANG ; Wei LI
Chinese Journal of Ultrasonography 2012;21(9):737-741
Objective To evaluate the right ventricular global systolic function by real-time three dimensional echocardiography (RT-3DE) and two dimensional echocardiography (2DE) in normal children,and investigate their correlation with age,height,weight,body surface area (BSA) and cardiac cycle time.Methods In 192 normal children,2DE right ventricular systolic function indexes were obtained at the apical four chamber view,including:tricuspid annular lateral wall peak systolic excursion (TAPSE),tricuspid annular peak systolic velocity (TRV) and right ventricular systolic area shortening (RVa).Full volume imaging of right ventricle was obtained at the parasternal four-chamber view near the apex by RT-3DE.The right ventricular ejection fraction (RVEF) was analyzed by TomTec RV-Function.The correlation analysis was performed between RT-3DE and 2DE indexes.The correlation and regression analysis on RT-3DE and 2DE indexes with above physical development indexes were also studied.At the same time 192 normal children were divided into five groups by age.2DE and RT-3DE indexes were compared with each other among five age groups.Results RVEF had no statistical correlation with all 2DE indexes (P >0.05).The correlation analysis on RT-3DE and 2DE indexes with physical development indexes demonstrated that TAPSE,TRV had strong positive correlation with the above indexes (P =0.000),while RVa had no statistical correlation with the above indexes (P >0.05).RVEF had weak positive correlation with cardiac cycle time r =0.180,P =0.014),but it had no statistical correlation with the other indexes (P >0.05).Stepwise regression analysis showed that the significant factor for TAPSE and TRV was height (R2 =0.508,P =0.000)and BSA (R2 =0.130,P =0.000)separately.In five age groups RVEF and RVa remained constant (P >0.05).However for TAPSE and TRV significant difference were found in part of comparisons between groups(P <0.05).Conclusions Although RVEF assessed by RT-3DE was slightly influenced by heart rate,it was independent of age,height,weight,BSA.It was a stable index for evaluating right ventricular systolic function among different age and physical development population.
8.Prevention of acute rejection of renal allograft in sensitized recipients
Yawang TANG ; Wen SUN ; Lei ZHANG ; Jun LIN ; Zelin XIE ; Ye TIAN
International Journal of Surgery 2010;37(1):43-46
objective To evaluate the influence of HLA matching and new immunosuppressants on pre-venting acute rejection of renal allograft in sensitized recipients. Methods 751 recipients underwent renal transplantation were enrolled in this study including 46 sensitized recipients (study group) with PRA be-tween 10%-90% and 705 non-sensitized recipients (control group) with PRA less than 10% pretransplant. All patients in the study group received induction course (ATG 100 mg/d, 5-7 d) plus triple-immunosup-pressive therapy including FK506 + MMF + steroid. The rate of acute rejection and delayed graft function after renal transplantation was analyzed. The influence of HLA matching on preventing acute rejection was al-so evaluated. Results The acute rejection rate in the study group and control group was 30.43% and 19. 57%, respectively, (P < 0.05). The rate of delayed graft function was 60.86% in the study group, signifi-cantly higher than that of the control group (11.87%). There was no statistically difference of one-year pa-tient / graft survival rotes between the two groups. The average serum creatinin levels at one-year posttrans-plantation were similar between the two groups (130 mmol/dl in the study group and 125 mmol/di in the control group). The average loci of HLA matching in the study group (4.2) was significantly higher than that in the control group (2.8). The acute rejection rate in the study group was significantly higher when lo-ci of HLA mismatch ranging from 2-4 compared with loci of HLA mismatch less than 2. The acute rejection rate was significantly higher in the highly sensitized recipients (PRA ranging from 50% -90% pretmnsplant) than that in the less sensitized (PRA ranging from 10% to 20% pretransplant) in the study group. Patients with higher PRA level posttransplantation were prone to developing acute rejection. Conclusion HLA matching and new immunosuppressants can reduce the incidence of acute rejection in sensitized recipi-ents and increase the survival rate of patients and allografts.
9.Randomised clinical trial on rh-Endo combined with FOLFOX4 regimen as an adjuvant therapy for stage Ⅱ and Ⅱ colorectal cancer patients
Zhihua XIE ; Dejian DAI ; Lin ZHONG ; Yi YI ; Jun FU ; Zhijin ZHANG ; Yuhao ZHANG
Chinese Journal of General Surgery 2013;28(10):758-762
Objective To evaluate the long-term efficacy of recombinant human endostatin (rh-Endo) combined with FOLFOX4 as an adjuvant treatment for patients of stage Ⅱ and Ⅲ colorectal cancer.Methods Eligible patients were randomly assigned to receive FOLFOX4 or FOLFOX4 plus rh-Endo regimen in which patients receiving 7.5 mg/m2 Ⅳ on day 1-7,repeated every 2 weeks,to a total of 12 cycles in 6 months.Results A total of 197 eligible patients were accrued in this research with 105 patients in the control group and 92 patients in the experimental arm.Median follow-up period was 42 months.The baseline characteristics distributed were balanced by treatment.Rh-Endo combined with FOLFOX4 regimen resulted in significant improvement on DFS compared to FOLFOX4 regimen for patients with stage Ⅲ colon cancer (HR =0.19,95% CI0.05-0.75,P =0.0124),and with a 34% improvement on 3-year DFS and 81% reduced recurrence.Although rh-Endo combined with FOLFOX4 regimen failed to make significant difference on DFS in the whole (HR =0.75,95% CI 0.31-1.83,P =0.5589),it was also observed a 17% improveiment on 3-year DFS.No statistical significant difference on DFS was observed in patients with stage Ⅱ disease.Conclusions Rh-Endo combined with FOLFOX4 regimen significantly improved the disease-free survival for patients with stage Ⅲ colorectal cancer,indicating that patients with stage Ⅲ disease,but not stage Ⅱ disease,can benefit from FOLFOX4 plus rh-Endo regimen in adjuvant treatment.
10.The safety and slow-release effect of chitosan-nanoparticle on the transforming growth factor-β receptor Ⅱ aptamer
Xia, CHEN ; Lei, LI ; Guang-jun, XIAN ; Wei, WANG ; Xiao-yan, ZHU ; Lin, XIE
Chinese Journal of Experimental Ophthalmology 2013;(4):352-357
Background Our previous study demonstrated that the aptamer S58 specifically targeted transforming growth factor-β receptor Ⅱ (TβRⅡ) and inhibited the transdifferentiation of human Tenon capsule fibroblasts (HTFs) mediated by transforming growth factor-β (TGF-β).Chitosan-nanoparticles (CS-NP) are good drug carriers,but the efficacy and safety of CS-NP/aptamer complexes deserve attention.Objective The aim of this study was to synthesize a novel CS-NP/aptamer complex called CS (S58)-NP and investigate its properties and applicability.Methods Human Tenon capsule tissue was obtained from patients during strabismus surgery,and HTFs were cultured and passaged using the explant culture method.The fourth to tenth generations of cells were used in the experiment.Different concentrations of CS-NP were used to prepare the CS(S58)-NP by the ionic cross-linking method with a surface charge rate (N/P) for S58 of 10,20,30 or 40.The particle size and Zeta potential were measured by the Zeta analyzer.The shape and distribution of CS (S58)-NP particles were examined under the scanning electron microscope.The binding of CS-NP with S58 and resistance of CS (S58)-NP to DNase Ⅰ were examined by agarose gel eletrophoresis.The release rate of S58 from CS (S58)-NP in PBS was quantitatively analyzed by a ultraviolet spectrophotometer.The cytotoxicity of CS(S58)-NP to HTFs was evaluated by detecting the production of lactate dehydrogenase (LDH).Results The Zeta analyzer showed that the particle size of CS (S58)-NP was 130-270 nm and its electric potential ranged from + 16 to +28 mV.The CS (S58)-NP particles appeared spherical with an even distribution under the scanning electron microscope.The mean encapsulation efficiency of CS(S58)-NP was 88.9%,89.3%,91.7% or 90.5%,respectively,when the N/P was 10,20,30 or 40.After being encapsuled by CS-NP,S58 could resist the degradation from DNase I.Its total releasing level in PBS increased with the lapse of time,with a maximum releasing speed at 24 to 36 hours.The total releasing level reached 100% at 96 hours.With increaseing concentrations of CS(S58)-NP,the relative releasing level of LDH in HTFs suspension gradually elevated with a significant difference among the groups (F =588.018,P =0.000),with the highest released LDH level at 50 nmol/L of CS(S58)-NP (12.853% ±0.375%).Conclusions CS-NP provides a protective and slow-releasing effect on the S58 aptamer.CS (S58)-NP shows a good biocompatibility with HTFs with a low cytotoxicity at a concentration of <50 nmol/L.CS(S58)-NP could be used to inhibit TGF-β induced transdifferentiation of HTFs in the future.