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Small intestinal submucosa improves islet survival and function during in vitro culture

World Journal of Gastroenterology 2005年第46期11卷 7378-7383页

作者： Xiao-Hui tian Wu-Jun Xue Xiao-Ming Ding Xin-Lu Pang Yan Teng pu-xun tian Xin-Shun Feng Center of Renal Transplantation The First Hospital of Xi'an Jiaotong UniversityXi'an 710061 Shaanxi Province China

AIM： To evaluate the recovery and function of isolated rat pancreatic islets during in vitro culture with small intestinal submucosa （SIS）. METHODS： Pancreatic islets were isolated from Wistar rats by standard surgical procurement followed by intraductal collagenase distension, mechanical dissociation and Euroficoll purification. purified islets were cultured in plates coated with multilayer SIS （SIS-treated group） or without multilayer SIS （standard cultured group） for 7 and 14 d in standard islet culture media of RPMI 1640. After isolation and culture, islets from both experimental groups were stained with dithizone and counted. Recovery of islets was determined by the ratio of counts after the culture to the yield of islets immediately following islet isolation. Viability of islets after the culture was assessed by the glucose challenge best with low （2.7 mmol/L） and high glucose （16.7 mmol/L） solution supplemented with 50 mmol/L 3-isobutyl-1- methylxanthine （IBMX） solution. Apoptosis of islet cells after the culture was measured by relative quantification of histone-complexed DNA fragments using ELISA. RESULTS： After 7 or 14 d of in vitro tissue culture, the recovery of islets in SIS-treated group was significantly higher than that cultured in plates without SIS coating. The recovery of islets in SIS-treated group was about twice more than that of in the control group. In SIS treated group, there was no significant difference in the recovery of islets between short- and long-term periods of culture （95.8±1.0% vs 90.8±1.5%, P〉0.05）. When incubated with high glucose （16.7 mmol/L） solution, insulin secretion in SIS-treated group showed a higher increase than that in control group after 14 d of culture （20.7±1.1 mU/L vs 11.8±1.1 mU/L, P〈0.05）. When islets were placed in high glucose solution containing IBMX, stimulated insulin secretion was higher in SlS-treated group than in control group. Calculated stimulation index of SlS-treated group was about 23 times of

关键词： Islet culture Islet survival Islet function Small intestinal submucosa

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Predictive Score Model for Delayed Graft Function Based on Hypothermic Machine Perfusion Variables in Kidney Transplantation

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Chinese Medical Journal 2018年第22期131卷 2651-2657页

作者： Chen-Guang Ding Yang Li Xiao-Hui tian Xiao-Jun Hu pu-xun tian Xiao-Ming Ding He-Li Xiang Jin Zheng Wu-Jun Xue Department of Kidney Transplantation Nephropathy HospitalThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anShaanxi 710061China Institute of Organ Transplantation Xi'an Jiaotong UniversityXi'anShaanxi 710061China

Background: Hypothermic machine perfusion(HMP) is being used more often in cardiac death kidney transplantation; however, the significance of assessing organ quality and predicting delayed graft function(DGF) by HMP parameters is still controversial. Therefore,we used a readily available HMP variable to design a scoring model that can identify the highest risk of DGF and provide the guidance and advice for organ allocation and DCD kidney ***: From September 1, 2012 to August 31, 2016, 366 qualified kidneys were randomly assigned to the development and validation cohorts in a 2:1 distribution. The HMP variables of the development cohort served as candidate univariate predictors for DGF. The independent predictors of DGF were identified by multivariate logistic regression analysis with a P < 0.05. According to the odds ratios(ORs) value, each HMP variable was assigned a weighted integer, and the sum of the integers indicated the total risk score for each kidney. The validation cohort was used to verify the accuracy and reliability of the scoring ***: HMP duration(OR = 1.165, 95% confidence interval [CI ]: 1.008–1.360, P = 0.043), resistance(OR = 2.190, 95%CI: 1.032–10.20, P < 0.001), and flow rate(OR = 0.931, 95% CI: 0.894–0.967, P = 0.011) were the independent predictors of identified DGF. The HMP predictive score ranged from 0 to 14, and there was a clear increase in the incidence of DGF, from the low predictive score group to the very high predictive score group. We formed four increasingly serious risk categories(scores 0–3, 4–7, 8–11, and 12–14)according to the frequency associated with the different risk scores of DGF. The HMP predictive score indicates good discriminative power with a c?statistic of 0.706 in the validation cohort, and it had significantly better prediction value for DGF compared to both terminal flow(P = 0.012) and resistance(P = 0.006).Conclusion: The HMP predictive score is a good noninvasive tool for assessing the qu

关键词： Delayed Graft Function Donation after Cardiac Death Hypothermic Machine Perfusion Kidney Transplantation

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Comparison of the characteristics of macrophages derived from murine spleen, peritoneal cavity, and bone marrow

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2017年第12期18卷 1055-1063页

作者： Yan-long ZHAO pu-xun tian Feng HAN Jin ZHENG Xin-xin XIA Wu-jun XUE Xiao-ming DING Chen-guang DING Department of Kidney Transplantation Hospital of Nephropathythe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'an 710061China Institute of Organ Transplantation Xi'an Jiaotong UniversityXi'an 710061China Department of Traditional Chinese Medicine the First Affiliated Hospital of Xi'an Jiaotong UniversityXi'an 710061China

Macrophages have a diverse set of functions based upon their activation states. The activation states, including resting（M0） and polarizing（M1 and M2） states, of macrophages derived from the mouse bone marrow, spleen, and peritoneal cavity（BMs, SPMs, and PCMs, respectively） were compared. We evaluated the macrophage yield per mouse and compared the surface markers major histocompatibility complex（MHC） Ⅱ and CD86 by flow cytometry. The relative mRNA levels of tumor necrosis factor-α（TNF-α）, interleukin（IL）-1β, mannose receptor（MR）, and Ym1 in the M0, M1, and M2 states were also compared using real-time polymerase chain reaction（PCR） analysis. Bone marrow yielded the most macrophages with the best homogeneity, but they were polarized toward the M2 phenotype. All three types of macrophages had the capacity to polarize into the M1 and M2 states, but SPMs had a stronger capacity to polarize into M1. The three types of macrophages showed no differences in their capacity to polarize into the M2 state. Therefore, the three types of macrophages have distinct characteristics regardless of their resting or polarizing states. Although bone marrow can get large amounts of homogeneous macrophages, the macrophages cannot replace tissue-derived macrophages.

关键词： Macrophage Murine splenocytes Murine bone marrow Peritoneal cavity

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Allograft rejection-related gene expression in the endothelial cells of renal transplantation recipients after cytomegalovirus infection

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第11期10卷 820-828页

作者： Yang LI Hang YAN Wu-jun XUE pu-xun tian Xiao-ming DING Xiao-ming PAN Xin-shun FENG Xiao-hui tian He-li XIANG Jun HOU Department of Renal Transplantation Center of Nephropathy the First Affiliated Hospital School of Medicine Xi 'an Jiaotong University Xi 'an 710061 China The two authors contributed equally to this work

Objective： To explore the effects of cytomegalovirus （CMV） infection on rejection-related gene expression in the endothelial cells of renal transplantation recipients. Methods： Endothelial cells （ECs） were cultured and stimulated by a variety of factors： A, normal control group; B, inactivated human cytomegalovirus （HCMV） infection group; C, HCMV infection group; D, HCMV supematant infection group; and E, ganciclovir HCMV group. Expression of intercellular adhesion molecule-1 （ICAM-1） and major histocompability complex （MHC） class I and class II antigens was detected by flow cytometry （FCM） and immuno- histochemistry. Results： We found characteristic CMV-infected ECs in this study. There were no significant differences among groups A, B and D （P〉0.05）. Although the expression levels of ICAM-1 were not significantly different between groups C and E （P〉0.05）, the ICAM-1 expression in these two groups was significantly higher than that in group A （P〈0.05）. ICAM-1 expression was detected in groups C and E, while there was no expression in groups A, B and D. Furthermore, there was no significant difference of ICAM-1 mRNA expression between groups C and E （P〉0.05）. Human leucocyte antigen （HLA）-ABC expression was detected in all the groups, while HLA-DR expression was only detected in groups C and E. There were no significant dif- ferences of HLA-ABC and HLA-DR expression among groups A, B and D （P〉0.05）. However, the HLA-ABC and HLA-DR expression levels in groups C and D were higher than those of the remaining groups previously reported （P〈0.05）. Meanwhile, the HLA-ABC and HLA-DR expression levels in group E were lower than those of group C （P〈0.05）. Conclusion： CMV could up-regulate the expression levels of ICAM-1 and MHC antigens, which was closely related to allograft rejection.

关键词： Renal transplantation Cytomegalovirus (CMV) Rejection Intercellular adhesion molecule-1 (ICAM-1) Major histocompability complex (MHC)

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Polyglycolic Acid Fibrous Scaffold Improving Endothelial Cell Coating and Vascularization of Islet

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Chinese Medical Journal 2017年第7期130卷 832-839页

作者： Yang Li Ping Fan Xiao-Ming Ding Xiao-Hui tian Xin-Shun Feng Hang Yan Xiao-Ming Pan pu-xun tian Jin Zheng Chen-Guang Ding Wu-Jun Xue Department of Renal Transplantation Center of Nephrology The First Affiliated Hospital of Xi'an Jiaotong University School of Medicine Xi'an Shaanxi 710061 China Department of Rheumatism and Immunology The First Affiliated Hospital of Xi'an Jiaotong University School of Medicine Xi'an Shaanxi 710061 China

Background：Improving islet graft revascularization has become a crucial task for prolonging islet graft *** cells （ECs） are the basis of new microvessels in an isolated islet,and EC coating has been demonstrated to improve the vascularization and survival of an ***,the traditional method of EC coating of islets has low efficiency in *** study was conducted to evaluate the effect of a polyglycolic acid （PGA） scaffold on the efficiency of islet coating by ECs and the angiogenesis in the coated islet ***：A PGA fibrous scaffold was used for EC coating of islet culture and was evaluated for its efficiency of EC coating on islets and islet graft ***：In in vitro experiments,we found that apoptosis index of ECs-coating islet in PGA group （27% ± 8%） was significantly lower than that in control group （83% ± 20%,P 〈 0.05） after 7 days *** index was significantly greater in the PGA group than in the control group at day 7 after ECs-coating （2.07 ± 0.31 vs.1.80 ± 0.23,P 〈 0.05）.vascular endothelial growth factor （VEGF） level in the PGA group was significantly higher than the coating in the control group after 7 days culture （52.10 ± 13.50 ng/ml vs.16.30 ± 8.10 ng/ml,P 〈 0.05）.Because of a tight,circumvallated,adhesive and three-dimensional growth microenvironment,islet cultured in a PGA scaffold had higher coating efficiency showing stronger staining intensity of enzyme than those in the control group after 14 days of culture following *** in vivo study,PGA scaffold significantly prolonged the average survival time of EC-coated islet graft after transplantation compared with control group （15.30 ± 5.60 days vs.8.30 ± 2.45 days,P 〈 0.05）.The angiogenesis and area of survived grafts were more in the PGA group compared with the control group by measuring the mean microvessel density （8.60 ± 1.21/mm2 vs.5.20 ± 0.87/mm2,P 〈 0.05）.In addition,expression of VEGF and tyrosin-protein kinase receptor

关键词： Endothelial Cells Islet Polyglycolic Acid Fibrous Revascularization

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Beneficial Effect of Moderately Increasing Hypothermic Machine Perfusion Pressure on Donor after Cardiac Death Renal Transplantation

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Chinese Medical Journal 2018年第22期131卷 2676-2682页

作者： Chen-Guang Ding pu-xun tian Xiao-Ming Ding He-Li Xiang Yang Li Xiao-Hui tian Feng Han Qian-Hui Tai Qian-Long Liu Jin Zheng Wu-Jun Xue Department of Renal Transplantation Nephropathy HospitalThe First Affiliated Hospital of Xi'an Jiaotong UniversityXi'anShaanxi 710061China Institute of Organ Transplantation Xi'an Jiaotong UniversityXi'anShaanxi 710061China

Background:Vascular resistance and flow rate during hypotherrnic machine perfusion (HMP)of kidneys is correlated with graft function. We aimed to determine the effects of increasing HMP pressure versus maintaining the initial pressure on kidney transplantation outcomes. Methods:We retrospectively reviewed the data of 76 primary transplantation patients who received HMP-preserved kidneys from 48 donors after cardiac death between September 1,2013,and August 31,*** pressure was increased from 30 to 40mmHg (1mmHg =0.133kPa)in kidneys with poor flow and/or vascular resistance (increased pressure [IP]group;36 patients);otherwise,the initial pressure was maintained (constant pressure group;40 patients).Finally,the clinical characteristics and transplantation outcomes in both groups were assessed. Results:Delayed graft function (DGF)incidence,1-year allograft,patient survival,kidney function recovery time,and serum creatinine level on day 30 were similar in both groups,with improved flow and resistance in the IP *** patients with DGF,kidney function recovery time and DGF duration were ameliorated in the IP *** logistic regression analysis revealed that donor hypertension (odds ratio [OR]:1.43,95%confidence interval [CI]:1.02-2.06,P =0.035),donor terminal serum creatinine (OR:1.27,95%C7:1.06-1.62,P =0.023),warm ischemic time (OR:3.45,95%CI:1.97-6.37,P =0.002),and terminal resistance (OR:3.12,95%CI:1.76-6.09,P =0.012)were independent predictors of *** proportional hazards analysis showed that terminal resistance (hazard ratio:2.06,95%C1:1.32-5.16,P =0.032)significantly affected graft survival. Conclusion:Increased HMP pressure improves graft perfusion but does not affect DGF incidence or 1-year graft survival.

关键词： Delayed Graft Function Donor after Cardiac Death Hypothermic Machine Perfusion Kidney Transplantation

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Early Immunosuppressive Exposure of Enteric-Coated-Mycophenolate Sodium Plus Tacrolimus Associated with Acute Rejection in Expanded Criteria Donor Kidney Transplantation

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Chinese Medical Journal 2018年第11期131卷 1302-1307页

作者： Chen-Guang Ding Li-Zi Jiao Feng Han He-Li Xiang pu-xun tian Xiao-Ming Ding Xiao-Ming Pan Xiao-Hui tian Yang Li Jin Zheng Wu-Jun Xue Department of Renal Transplantation Nephropathy Hospital The First Affiliated Hospital of Xi'an Jiaotong University Xi'an Shaanxi 710061 China Institute of Organ Transplantation Xi'an Jiaotong University Xi'an Shaanxi 710061 China

Background： lmmunosuppressive agents are still inefficient in preventing biopsy-proven acute rejection （BPAR） alter expanded criteria donor （ECD） kidney transplantation. The aim of this study was to investigate the relationships between early imrnunosuppressive exposure and the development of BPAR. Methods： We performed a retrospective study of 58 recipients of ECD kidney transplantation treated with enteric-coated-mycophenolate sodium, tacrolirnus （Tac）, and prednisone. The levels of mycophenolic acid-area under the curve （MPA-AUC）0-12h and Tac C0 were measured at the 1st week and the 1st month posttransplant, respectively. The correlation was assessed by multivariate logistic regression. Results： The occurrence rates of BPAR and antibody-mediated rejection were 24.1% and 10.3%, respectively. A low level of MPA-AUC0-12h at the 1st week posttransplant was found in BPAR recipients （38.42 ± 8.37 vs. 50.64 ± 13.22, P 〈 0.01）. In addition, the incidence of BPAR was significantly high （P 〈 0.05） when the MPA-AUC0-12h level was 〈30 mg·h-1·L-1 at the 1st week （ 15.0% vs. 44.4%） or the Tac C0 was 〈4 ng/ml at the 1 st month posttransplant （33.3% vs. 21.6%）. Multivariable logistic regression analysis showed that the MPA-AUC 0-12 h at the 1st week （OR： 0.842, 95% CI： 0.784-0.903） and the Tac C0 at the 1st month （OR： 0.904, 95% C7： 0.822-0.986） had significant inverse correlation with BPA R （ P 〈 0.05 ）. Conclusions： Low-level exposure of MPA and Tac C0 in the early weeks posttransplant reflects an increased acute rejection risk, which suggested that MPA-AUC0-12h 〈30 mg·h-1·L -1 and Tac C0 〈4 ng/ml should be avoided in the first few weeks alter transplantation.

关键词： Enteric-Coated-Mycophenolate Sodium Tacrolimus Acute Rejection Expanded Criteria Donor Kidney Transplantation

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Predictive Score Model for Delayed Graft Function Based on Easily Available Variables before Kidney Donation after Cardiac Death

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Chinese Medical Journal 2017年第20期130卷 2429-2434页

作者： Chen-Guang Ding Qian-Hui Tai Feng Han Yang Li Xiao-Hui tian pu-xun tian Xiao-Ming Ding Xiao-Ming Pan Jin Zheng He-Li Xiang Wu-Jun Xue Department of Renal Transplantation Nephropathy Hospital The First Affiliated Hospital of Xi'an Jiaotong University Xi'an Shaanxi 710061 China Institute of Organ Transplantation Xi'an Jiaotong University Xi'an Shaanxi 710061 China

Background： How to evaluate the quality of donation after cardiac transplantation in China. Hence, the aim of this study was to develop kidneys before DCD. death （DCD） kidneys has become a critical problem in kidney a simple donor risk score model to evaluate the quality of DCD Methods： A total of 543 qualified kidneys were randomized in a 2：1 manner to create the development and validation cohorts. The donor variables in the development cohort were considered as candidate univariate predictors of delayed graft function （DGF）. Multivariate logistic regression was then used to identify independent predictors of DGF with P 〈 0.05. Date from validation cohort were used to validate the donor scoring model. Results： Based on the odds ratios, eight identified variables were assigned a weighted integer; the sum of the integer was the total risk score for each kidney. The donor risk score, ranging from 0 to 28, demonstrated good discriminative power with a C-statistic of 0.790. Similar results were obtained from validation cohort with C-statistic of 0.783. Based on the obtained frequencies of DGF in relation to different risk scores, we formed tour risk categories of increasing severity （scores 04, 5 9, 10-14, and 15 28）. Conclusions： The scoring model might be a good noninvasive tool for assessing the quality of DCD kidneys before donation and potentially useful for physicians to make optimal decisions about donor organ offers.

关键词： Delayed Graft Function Donation after Cardiac Death Kidney Transplantation Predictive Score

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Magnetic cell sorting and flow cytometry sorting methods for the isolation and function analysis of mouse CD^4+ CD25^+ Treg cells

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第12期10卷 928-932页

作者： Hang YAN Chen-guang DING pu-xun tian Guan-qun GE Zhan-kui JIN Li-ning JIA Xiao-ming DING Xiao-ming PAN Wu-jun XUE Department of Renal Transplantation Center of Nephropathy the First Affiliated Hospital Medicine College Xi'an Jiaotong University Xi'an 710061 China

Objective: In this paper we compared the two methods of cell sorting (magnetic cell sorting and flow cytometry sorting) for the isolation and function analysis of mouse CD4+ CD25+ regulatory T (Treg) cells, in order to inform further studies in Treg cell function. Methods: We separately used magnetic cell sorting and flow cytometry sorting to identify CD4+ CD25+ Treg cells. After magnetic cell separation, we further used flow cytometry to analyze the purity of CD4+ CD25+ Treg cells, trypan blue staining to detect cell viability, and propidium iodide (PI) staining to assess the cell viability. We detected the immune inhibition of CD4+ CD25+ Treg cells in the in vitro proliferation experiments. Results: The results showed that compared to flow cytometry sorting, magnetic cell sorting took more time and effort, but fewer live cells were obtained than with flow cytometry sorting. The CD4+ CD25+ Treg cells, however, obtained with both methods have similar immunosuppressive capacities. Conclusion: The result suggests that both methods can be used in isolating CD4+ CD25+ Treg cells, and one can select the best method according to specific needs and availability of the methodologies.

关键词： CD4+ CD25+ Treg cells Flow cytometry sorting Magnetic cell sorting

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The clinical value of enzyme-multiplied immunoassay technique monitoring the plasma concentrations of cyclosporine A after renal transplantation

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Journal of Pharmaceutical Analysis 2011年第2期1卷 139-142页

作者： Xiao-Hui Luo wu-Jun Xue pu-xun tian Xiao-Ming Ding Hang Yan He-Li Xiang Yang Li Department of Renal Transplant Center of Nephropathy the First Affiliated Hospital Medical School of Xi'an Jiaotong University Xi'an 710061 China.

The feasibility and the clinical value of the enzyme-multiplied immunoassay technique （EMIT） monitoring of blood concentrations of cyclosporine A （CsA） in patients treated with CsA were investigated after kidney transplantation. The validation method was performed to the EMIT determination of CsA blood concentration, the CsA whole blood trough concentrations （Co） of patients in different time periods after renal transplantation were monitored, and combined with the clinical complications, the statistical results were analyzed and compared. EMIT was precise, accurate and stable, also with a high quality control. The mean postoperative blood concentration of CsA was as follows：〈1 month, （281.4± 57.9）ng/mL; 2 - 3 months, （264.5 ± 41.2） ng/mL; 4 - 5 months, （236.4 ± 38.9） ng/mL; 6 - 12 months, （206.5± 32.6）ng/mL; 〉12 months, （185.6± 28.1）ng/mL. The toxic reaction rate of CsA blood concentration within the recommended therapeutic concentration was 14.1%, significantly lower than that of the none-recommended dose group （37.2%）（P〈0.05）; the transplantation rejection rate was 4.4%, significantly lower than that of the none- recommended dose group （22.5%）（P〈0.05）. Using EMIT to monitor the blood concentration of CsA as the routine laboratory method is feasible, and is able to reduce the CsA toxicity and rejection significantly, leading to achieving the desired therapeutic effect.

关键词： enzyme-multiplied immunoassay technique renal transplantation cyclosporin A blood concentration monitoring

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