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.Infection and its prophylaxis in renal transplanted patients after Basiliximab induction therapy
Linlin MA ; Zelin XIE ; Yawang TANG ; Ye TIAN ; Wen SUN ; Hongbo GUO ; Jun LIN ; Lei ZHANG
Chinese Journal of Organ Transplantation 2011;32(4):205-208
Objective To investigate the incidence of infection and the effect of anti-infection prophylaxis in renal transplanted patients after Basiliximab induction therapy. Methods A total of 204patients who have received renal transplantation and Basiliximab induction therapy from January 1,2001 to December 31, 2010 in our hospital have been retrospective analysed in this study. These patients were divided into a prophylaxis group (118 cases) with Ganciclovir + Sulfadiazine +Trimethoprim therapy and a control group (86 cases) without any anti-infection prophylaxis.Furthermore, 440 transplanted patients in the same peroid without any induction therapy were also analysed. They were also devided into two groups: an anti-infection prophylaxis group (206 cases)and a control group (234 cases) without any anti-infection prophylaxis. Results In the prophylaxis group with Basiliximab induction therapy, there were 23 patients (19. 5 %, 23/118)experienced hospitalization due to infection, 3 cases (13. 0 %,3/23) among them were severe infection, and 3patients (13.0 %, 3/23) died from vital infection. In the non-prophylaxis control group with Basiliximab induction therapy, 27 patients (31.4 %, 27/86) had infection complication, 7 patients (25.9 % ,7/27) among them were severe infection, and 4 patients(14. 8 % ,4/27)died. The incidence of infection between the above two groups is significantly different (P<0. 05). In the prophylaxis group without induction therapy, the incidence of infection was 15.0 % (31/206), there were no severe infection cases but 7 patients (22. 6 %, 7/31) died from infection. In the non-prophylaxis control group without induction therapy, the incidence of infection was 12. 8 % (30/234), 3 cases among them were severe infection(10. 0 %,3/30)and 5 patients died from infection (16. 7 %, 5/30).The incidence of infection in Basiliximab induced patients without anti-infection prophylaxis is significantly higher than that in patients without induction therapy and anti-infection prophylaxis (31.4 % vs. 12.8 %,P<0.01). Conclusion Basiliximab induction therapy increased the risk of infection, but not the rate of mortality. It is necessary to give anti-infection prophylaxis in renal transplanted patients with Basiliximab induction therapy.
7.Evaluation of right ventricular global and regional volume and systolic function in normal children by real-time three-dimensional echocardiography
Jun GAO ; Mingxing XIE ; Qing Lü ; Lingyun FANG ; Lin HE ; Li ZHANG ; Wei LI
Chinese Journal of Ultrasonography 2011;20(6):461-465
Objective To evaluate right ventricular(RV) global and regional volume and systolic function by real-time three-dimensional echocardiography (RT-3DE) in normal children with different age.Methods One hundred and ninty-two normal children were divided into five groups by age:group Ⅰ,<1 years old,32 cases;group Ⅱ,≥1 years old-<3 years old,46 cases;group Ⅲ,≥3 years old-<6 years old,36 cases;group Ⅳ,≥6 years old-<9 years old,41 cases;group Ⅴ,≥9 years old-<14 years old,37 cases.Full volume imaging of RV was obtained at the parasternal four-chamber view near the apex by RT-3DE.RT-3DE data set were analyzed off-line by TomTec RV-Function.RV were divided into three parts:inflow,body,outflow.The measurements were including RV global and regional end-diastolic volume (EDV),end-systolic volume (ESV),ejection fraction (EF) and the ratio of regional parts EDV to RVEDV. The volume and systolic function were compared in three regional parts.RV global and regional parts EF and the ratio of regional parts EDV to RVEDV were also compared in five age groups.Correlation analysis and curve estimation were studied on RV global and regional EDV with age and physical development indexes.Results In the comparison of three regional parts:inflow EDV and EF were higher than outflow and body parts (P<0.05).No significant different was found between outflow EDV and body EDV (P>0.05),however,outflow EF was significant higher than body EF(P<0.05).The comparison of RV global and regional EF in five age groups were no statistical different (P>0.12). The ratio of regional parts EDV to RVEDV remained constant in five age groups(P>0.58).Correlation analysis showed the global and regional RV volume were strongly correlated with age,height,weight and BSA (r>0.77,P=0.000).The best correlation was found with BSA (r>0.83,P=0.000).Curve estimation demonstrated that the relationship of RV global and regional EDV with age and physical development indexes could be best expressed by power model,the best matched model were found with BSA.Conclusions Among three regional parts of RV,inflow and outflow parts volume contraction were the two main contribution factors for RV function.In childhood RV volume didn't increase linearly with age and physical development indexes,but in an exponential model.
8.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.
9.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.
10.Effect of Okam on Airway Inflammation in Asthmatic Mouse
zheng-hai, QU ; ning, XIE ; xiao-mei, LIU ; rong-jun, LIN
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To explore the effect of Okam on airway inflammation in asthmatic mouse.Methods Thirty-two SPF grade Kunming Strain mice were randomly divided into positive control group,glucocorticoid inhalation group,Okam group and negative control group with 8 mice in each group.The mice were sensitized and repeatedly challenged with ovalbumin(OVA) to establish the models of chronic asthma.The glucocorticoid group were given Budesonide(200 ?g) and saline everyday by inhalation,the Okam group were given 50 mg/kg Okam by gavage,and the positive group had saline at the same time,the negative control group received saline at all stages.The inflammation of the lung tissue were scored underwent HE staining.Bronchoalveolar lavage fluid(BALF) cell count and differential were studied,and interferon-?(IFN-?),interleukin-4(IL-4) in BALF were determined by enzyme-linked immunosorbent assay(ELISA).Results There were no inflammatory cell infiltrate of bronchiole in the negative control group.Inflammatory infiltration of lung tissue were obvious in the positive control group.Inflammatory infiltration of lung tissue lightened obviously in the Budesonide and Okam groups.The total cell number,Eosinophils(EOS) and IL-4 level in BALF,and the score of the lung tissue in Okam group were all markedly lower than those in positive control group(t=5.942,7.089,7.078 Pa0.05),IFN-? lower(t=4.275 P