1.Expression of HLA-G5 in healthy people and the recipients of renal and liver transplantation
Li XIAO ; Bingyi SHI ; Ming CAI
Medical Journal of Chinese People's Liberation Army 1982;0(01):-
Objective To investigate the expression of human leukocyte antigen-G5 (HLA-G5) in healthy Chinese people and the recipients of renal and liver transplantation. The regulating mechanism of the expression of HLA-G5 was discussed by comparing the expression of HLA-G5 in the healthy people with that in renal and liver transplantation recipients. Furthermore, the changing regularity with time was studied by kinesis supervising the expression of HLA-G5 in renal and liver transplantation recipients. Methods The peripheral blood samples (3ml) from 30 health people, 50 recipients of liver transplantation (liver function was stable 3 months after liver transplantation) and 50 recipients of renal transplantation (renal function was stable 3 months after renal transplantation) were collected. Peripheral blood samples were also collected in same amount from 33 recipients of renal and liver transplantation before operation and 1, 4 and 12 weeks and 1 year after operation. The HLA-G5 of all serum samples was analyzed by ELISA. Results For 30 healthy people, the OD value of HLA-G5 in 28 people was below 0.5, for which the contents were defined as 0.0ng/ml according to standard and the contents for the other 2 people were 8ng/ml and 9ng/ml, respectively. 16 of 50 recipients undergone liver transplantation were positive for the expression of HLA-G5, the positive ratio was 32%. The contents in 4 recipients were higher than 30ng/ml. 10 of 50 recipients of renal transplantation were positive in the expression of HLA-G5, the positive ratio was 20%. The contents in one recipient were higher than 25ng/ml. The average contents in sera of healthy people, recipients of liver or renal transplantation were 0.56?0.20ng/ml, 8.34?1.50ng/ml and 3.26?0.25ng/ml, respectively. For 33 recipients of liver or renal transplantation, the expression of HLA-G5 was detected by ELISA, and it was found that one recipient the expression of HLA-G5 was positive before operation and within 1 week after operation; expression of HLA-G5 was positive in 4 recipients within 4 weeks after operation; expression of HLA-G5 was positive within 12 weeks after operation in 12 recipients; and the expression of HLA-G5 was positive within 1 year after operation in 11 recipients. Conclusion The expression of HLA-G5 in healthy people is low. There are correlation between the expression of HLA-G5 and immunotolerance to transplants. In minor rejection condition after transplantation, there are different expression levels of HLA-G5, and it is higher after liver transplantation than!renal transplantation. The time for expression of HLA-G5 corresponds with the time for mRNA of HLA-G5 transcription into protein, and it is about 15-60 days, with 60 days as the peak time.
2.Role of 5-hydroxy trptamine 2A receptor in biliary fibrosis after liver transplantation in rats
Liping CHEN ; Yawen WANG ; Li XIAO ; Wenmei FAN ; Bingyi SHI
Chinese Journal of Organ Transplantation 2014;35(9):555-560
Objective To evaluate the role of 5-hydroxy trptarine 2A receptor (5-HT2A) in the pathogenesis of biliary fibrosis after liver transplantation in rats.Method Rats were randomly divided into control group Ⅰ and control group Ⅱ[(supplied livers were preserved for 1 or 12 h),ketanserin group (recipients of control group Ⅱ were intraperitoneally injected with ketanserin 24 h postoperatively at the dosage of 5 mg · kg-1 · day-1),and sham group (rats were subjected to transverse laparotomy and closure without manipulation of the liver).During 4-week observation period,serum biliary enzymes,5-HT content in the liver,the expression of fibrosis-related genes,cholangiocytes proliferation and biliary fibrosis were evaluated.Result Compared with the sham group,the serum ALP,GGT,TBil and 5-HT contents in the liver homogenate were increased on the postoperative day 1 (POD1) and then restored to the normal level.There was slight proliferation of bile duct epithelial cells on POD3 in the control group Ⅰ,with fewer collagen fibers and α-sooth muscle actin (α-SMA)-positive myofibroblasts in the portal area.The expression of matrix metalloproteinase-2 (MMP-2) and procollagen α1-mRNA in graft livers was not significantly increased in the control group Ⅰ.To the contrast,the control group Ⅱ demonstrated high levels of serotonin in the liver homogenate and enhanced serum biliary enzymes.Active cholangiocytes proliferation was triggered on POD3 and remained higher than in the control group Ⅰ and the sham group.The control group Ⅱ showed a large number of α-SMA-positive myofibroblasts and collegan fibers at the postoperative week 4.In parallel,the major profibrogenic transcripts MMP2 and procollagen α1 were significantly increased at 2nd,and 4th week postoperation in the control group Ⅱ.Importantly,we also found that ketanserin relieved the signs of biliary fibrosis at 4th week postoperation in 5-HT2A group by the demonstration of reduced collagen fibers and a-SMA-positive myofibroblast in the portal area,as well as the decrease in the fibrosis-related gene expression.In addition to the lower cholangiocytes proliferation,serum levels of biliary enzymes including GGT,ALP and TBil in 5-HT2A group were significantly decreased at 4th week postoperation as compared with the control group Ⅱ.Conclusion Selective 5-HT2A receptor antagonist,Ketanserin retards biliary fibrosis progression posttransplantation,suggesting that 5-HT2A receptor is a potential therapeutic target for ischemia-related biliary fibrosis after DCD liver transplantation.
3.Clinical characteristics and management of acute myocardial infarction after kidney transplantation
Liping CHEN ; Xiaoling ZHANG ; Li XIAO ; Yeyong QIAN ; Bingyi SHI
Chinese Journal of Organ Transplantation 2013;34(10):604-606
Objective To explore the clinical characteristics and management of acute myocardial infarction (AMI) early after kidney transplantation (<3 months).Method Five cases of AMI early posttransplantation among 122 kidney transplant recipients from June 2011 to December 2012 were retrospectively reviewed.Results Of 5 AMI patients,there were 2 cases within one week postoperatively,one case at 11 th day postoperation,and the other two at 29th day and 46th day after operation respectively.Acute left heart failure was complicated in 3 cases within first two weeks.All the AMI patients had elevated TnⅠ levels which declined subsequently.The climax of TnⅠ levels in all the 5 AMI patients were above 5 ng/mL,and more than 20 ng/mL in two AMI patients within one week.Given by symptomatic and supportive treatment,antiplatelet and anticoagulation therapies and cardioprotective medications,all the five AMI patients were improved.Low molecular heparin was additionally administrated to the 2 cases within first week according to the severe conditions.New emerged small volume of hematocele was proved by ultrasound after 3 days and low molecular heparin was ceased.All the 5 patients survived and neither thrombolysis nor percutaneous coronary intervention therapy was given to them.Conclusion In addition to general prevention against AMI in kidney recipients with high risk factors,managing anemia and hypertensiorn,and improving graft function and systematic status are also important to decrease the risk of AMI.Moreover,cardioprotective therapy including antiplatelet therapies,beta-blockers,angiotensin-converting enzyme inhibitors (ACEI)/angiotensin-2 receptor blockers and statins,which are recommended to the general population with AMI,will also profit to the kidney transplant recipients with AMI.However,aggressive intervention therapies might be more prudent to be used in this population.
4.Human CMV Infection and Change in T Lymphocyte Subpopulation in Organ Recipients
Jian BAI ; Bingyi SHI ; Li XIAO ; Meng TANG ; Xiangke PEI ; You LIU
Chinese Journal of Nosocomiology 2006;0(09):-
0.05).The CD4+ lymphocyte of them was(53.37?4.38)% and(29.25?9.38)%,respectively,and the difference between two groups was significant(P
5.Expression and significance of soluble LAIR in sera of renal transplantation recipients with cytomegalovirus pneumonitis
Yubo ZHAO ; Chunyan WANG ; Bingyi SHI ; Boquan JIN ; Li XIAO ; Ruipeng HOU ; Zhouli LI
Chinese Journal of Urology 2008;(11):752-754
Objective To study the relationship of soluble LAIR (sCD305 and CD3060) expression in recipient serum with cytomegalovirus (CMV) pneumonitis after renal transplantation. Methods Nineteen serum specimens from recipients were divided into CMV pneumonitis group (n=10) and control group (n=9). Then the concentrations of sCD305 and CD3060 were quantitated with sandwich ELISA. The data were analyzed by using student t test. Results sCD305 was skewness distributed in both 2 groups, was 0.000-3.039 μg/L in CMV pneumonitis group and 0.000-8.375 μg/L in con-trol group. CD3060 was skewness distributed in CMV pneumonitis group and the concentration was 0.000-0.017μg/L. CD3060 was mormally distributed in control group and the concentration was 0.046±0.035 μg/L. There was significant difference of CD3060 (P=0.000) concentrations and no sig-nificant difference of sCD305(P=0.316) concentrations in 2 groups, respectively. Conclusions The concentration of CD3060 is low in CMV pneumonitis patients. The combination of CMV PP65 antigen detection and CD3060 detection is helpful for the early and precise diagnosis of CMV pneumonitis in renal transplantation patients.
6.Isolation and biological characteristics of rat umbilical cord mesenchymal stem cells
Kuili LIU ; Bingyi SHI ; Dezhong LIU ; Jiangang JIN ; Haibin LI ; Yingchang SHI ; Kai FENG ; Li XIAO
Chinese Journal of Tissue Engineering Research 2010;14(10):1743-1748
BACKGROUND:There are many studies concerning rat bone marrow mesenchymal stem cells for immune tolerance following transplantation and tissue repair.However,there are no reports on umbilical cord mesenchymal stem cells(UCMSCs).OBJECTIVE:To establish a method of separating mesenchymal stem cells(MSCs)from rat umbilical cord,and to study its biological characteristics.METHODS:MSCs were separated from rat umbilical cord with enzyme method and tissue mass method,and then incubated in DMEM-LG medium.Cell morphology was observed under an inverted microscope.Growth curves of cells were drawn using cell counting.Cell cycle and surface antigen were detected with flow cytometry.Adipogenic differentiation and osteogenic differentiation were tested by immunohistochemistry.RESULTS AND CONCLUSION:Both of the two methods could obtain plenty of MSCs from rat umbilical cord.Primary culture showed that the efficiency of enzyme method was higher than tissue mass method.Passage time of the former was about 10 days and the latter was 14 days.The passage time of latter except primary culture was the same.Immunophenotype analysis showed that MSCs from rat umbilical cord expressed adhesion molecule and stromal cell markers,CD90 and CD106,but did not express hematopoietic cell markers,CD34 and CD45.In vitro induction test verified that rat UCMSCs have the potentials of adipogenic and osteogenic differentiation.
7.Clinical study of Simulect in renal allograft recipients
Xuren XIAO ; Jianhua AO ; Zhilian MIN ; Xiaoda TANG ; Bingyi SHI ; Jianyu LING
Chinese Journal of Urology 2001;0(11):-
Objective To evaluate the efficacy and safety of Simulect for the prevention of acute rejection in renal allograft recipients receiving Neoral-based immunosuppressive regimen. Methods A prospective,multicenter and open-label clinical trial was conducted from March to October 2001.A total of 33 patients [20 men and 13 women; age range,18-63 years;mean age,(42.6?11.6) years] who received first kidney allograft were enrolled.Thirty-two cases had panel-reactive antibody
8.Determination of cytokines in peripheral blood by cytometric bead array in kidney transplantation recipients
Li XIAO ; Bingyi SHI ; Xiuyun HE ; Xiaoguang XU ; Yong HAN ; Wenqiang ZHOU ; Haiyan HUANG ; Yu GAO
Chinese Journal of Laboratory Medicine 2010;33(12):1128-1132
Objective To measure the cytokines levels in peripheral blood from kidney transplantation recipients by using cytometric bead array and to analyze their change and the clinical significance in pre- and post- kidney transplantation, inducting with basiliximab and graft rejection. Methods A total of 72 renal transplantation recipients were divided into two groups, kidney function stable group(n =53) and acute rejection group (n = 19). And they were also grouped by induction with basiliximab or not,32 in basiliximab group and 40 in without basilixmab group. The levels of IFN-γ, TNF-α, IL-10, IL-5,IL-4, IL-2 were measured by cytometric bead array in peripheral blood of 72 kidney transplantation recipients and 30 healthy donors at differential time. The data was analyzed according to the following grouping:donors and recipients, kidney function stable group and acute rejection group post transplantation and with or without basiliximab group. Results The levels of TNF-α, IL-10, IL-5, IL-4, IL-2 in recipients before transplantation were ( 1.65 ±0. 10) ,(2. 55 ±0. 19) ,( 1.88 ±0. 14) ,(1.85 ±0. 12) ,(2. 12 ±0. 09) ng/L,respectively. While they were (3.04 ±0. 17), (3.33 ±0. 26), (4.03 ±0.25), (2.73 ±0. 16), (4.03 ±0. 26) ng/L respectively in healthy donors. There was statistical significance between the two groups ( t =6. 890, 2. 375, 7. 851,3.955,7.153, P<0. 01, <0. 05, <0.01, <0.01, <0.01). While the level of IFN-γ in recipients before transplantation was (2. 50 ±0. 18) ng/L,compared with (3. 00 ±0. 24) ng/L in healthy donors. There was no statistical significance between the two groups( t = 1. 625, P > 0. 05 ). The levels of IFN-γ and IL-10 in kidney function stable group were (2. 71 ± 0. 11 ) ng/L and (3.91 ± 0. 52) ng/L,while they were ( 3.30 ± 0. 36 ) ng/L and ( 12. 01 ± 5.35 ) ng/L in acute rejection group. There were statistical dirrerences between the two groups ( t = 5. 061, 11. 465, P < 0. 01, < 0. 05 ). Before induction with basiliximab, the levels of IFN-γ, TNF-α, IL-10 in recipients were (2.90 ±0. 21 ), ( 1.67 ±0. 12),(2. 45 ± 0. 16) ng/L respectively. But they were ( 2. 78 ± 0. 17 ), ( 1.58 ± 0. 07 ), ( 2. 77 ± 0. 24 ) ng/L respectively after induction with basiliximab, which showed significantly different ( t = 5. 605, 6.011,4. 126, P <0. 01, <0. 01, <0. 05). Four weeks after kidney transplantation in recipients with basiliximab,the levels of IFN-γ, IL-10, IL-4 were (2. 90 ± 0. 31 ), (9. 08 ± 0. 16), (2. 73 ± 0. 11 ) ng/L. While they were (3.28 ±0. 11 ), (4. 17 ±0. 21 ), (2. 11 ±0. 20) ng/L respectively in recipients without basiliximab induction, which were significantly different from those with basiliximab induction (t = 4. 268,4. 263,3.762, P <0. 01, <0. 01, < 0. 05 ). Conclusions Six kinds of cytokines can be measured by cytometric bead array simultaneously and accurately. The data suggests that the detection of multiple cytokines in kidney transplantation recipients by cytometric bead array can provide more guidance for clinical diagnosis and therapy.
9.Correlation of HLA-G expression with CMV active infection following kidney transplantation
Li XIAO ; Bingyi SHI ; Yu GAO ; Xiuyun HE ; Xiaoguang XU ; Haiyan HUANG ; Wenqiang ZHOU ; Yong HAN
Chinese Journal of Organ Transplantation 2011;32(9):534-538
ObjectiveTo determine the correlation of human leukocyte antigen-G (HLA-G)expression with CMV active infection after kidney transplantation. MethodsA total of 215 first-time kidney transplantation recipients in one transplantation center were divided into CMV ( + ) group and CMV ( - ) group according to whether they had active CMV infection. mhla-g1 expression on leukocytes was analyzed by flow cytometry. The concentrations of soluble HLA-G5 were detected by using ELISA. The sHLA-G5 cutoff levels by ROC curve was employed to predict the active CMV infection. The expression of sHLA-G5 mRNA and protein in leukocytes was analyzed by using RTPCR and Western blotting respectively. Immunohistochemical staining was used to detect the HLA-G expression in kidney biopsies of 12 cases. ResultsThe expression of mHLA-G1 in peripheral blood was low in both CMV ( + ) group and CMV ( - ) group. Also when CMV-PP65 was positive, there was no significant change in mHLA-G1. In CMV ( + ) group, the proportion of CD14+ mHLA-G1 +cells[(45. 53 ± 17.32)%]in peripheral blood was increased as compared with that in CMV (-)group[(10. 22 ± 5.78)%]. The expression of sHLA-G5 was increased significantly in CMV ( + )group. The optimal cutoff value of sHLA-G5 predicting the active CMV infection was 202. 9 μg/L,with high diagnostic accuracy. HLA-G was positive in the kidney tissue of 10 patients out of 12 patients with active CMV infection. Both RT-PCR and Western blot analysis showed that sHLA-G5 was significantly higher in CMV ( + ) group than that in CMV ( - ) group. ConclusionROC curve analysis of sHLA-G5 with the cutoff value of 202. 9 μg/L can be used to predict the active CMV infection. The HLA-G levels in peripheral blood were significantly increased and HLA-G expression in the tubular epithelial cells of the graft could be a protection mechanism of the kidney function.
10.The correlation of HLA-G expression with AR and CMV active infection after kidney transplantation
Li XIAO ; Bingyi SHI ; Yu GAO ; Xiuyun HE ; Xiaoguang XU ; Haiyan HUANG ; Wenqiang ZHOU ; Yong HAN
Chinese Journal of Organ Transplantation 2011;32(10):584-587
Objective To study the correlation of HLA-G levels with acute rejection and CMV active infection post-kidney transplantation.Methods A total of 132 initial kidney transplantation recipients were divided into kidney function stable group (F),acute rejection group (AR),CMV group according to whether they had active CMV infection and acute rejection.Forty-one healthy donors served as control group (H).HLA-G levels and mRNA expression were analyzed by using flow cytometry,ELISA,RT-PCR and Western blotting.Immunohistochemical staining was used to detect the HLA-G expression in kidney biopsies.Results The expression levels of mHLA-G1 were low in all 4 groups pre-transplantation.Only CMV group had significantly more CD14+ mHLA-G1+ cells post-transplantation (P<0.05).sHLA-G5 levels were higher in F group than in H group (P<0.05),but there was no significant difference among other groups pre-transplantation (P>0.05).sHLA-G5 levels were increased significantly in CMV group as compared with F group (P<0.05),and those in F group were higher than in H and AR groups (P<0.05).Renal tissue biopsies from 21 renal transplantation recipients with AR indicated that HLA-G5 was expressed negatively in 17 patients,positively in 3 patients and 1 weakly positively.HLA-G was positive in the kidney tissue of 9 patients out of 9 patients with active CMV infection.In total 132 recipients,AR incidence was significantly lower in CMV ( + ) group (7.1 %,2/28) than that in CMV ( - ) group (24.0 %,25/104).Conclusion The sHLA-G5 may contribute to predict AR and CMV active infection; AR and CMV active infection may be correlation with immune balance in kidney transplantation recipients.