Donor-specific antibodies, glomerulitis, and human leukocyte antigen B eplet mismatch are risk factors for peritubular capillary C4d deposition in renal allografts / 中华医学杂志(英文版)

BACKGROUND@#The complement system plays an important role in the immune response to transplantation, and the diagnostic significance of peritubular capillary (PTC) C4d deposition (C4d+) in grafts is controversial. The study aimed to fully investigate the risk factors for PTC C4d+ and analyze its significance in biopsy pathology of kidney transplantation.@*METHODS@#This retrospective study included 124 cases of kidney transplant with graft biopsy and donor-specific antibody (DSA) testing from January 2017 to December 2019 in a single center. The effects of recipient pathological indicators, eplet mismatch (MM), and DSAs on PTC C4d+ were examined using univariate and multivariate logistic regression analyses.@*RESULTS@#In total, 35/124 (28%) were PTC C4d+, including 21 with antibody-mediated rejection (AMR), eight with renal tubular injury, three with T cell-mediated rejection, one with glomerular disease, and two others. Univariate analysis revealed that DSAs (P < 0.001), glomerulitis (P < 0.001), peritubular capillaritis (P < 0.001), and human leukocyte antigen (HLA) B eplet MM (P = 0.010) were the influencing factors of PTC C4d+. According to multivariate analysis, DSAs (odds ratio [OR]: 9.608, 95% confidence interval [CI]: 2.742-33.668, P < 0.001), glomerulitis (OR: 3.581, 95%CI: 1.246-10.289, P = 0.018), and HLA B eplet MM (OR: 1.166, 95%CI: 1.005-1.353, P = 0.042) were the independent risk factors for PTC C4d+. In receiver operating characteristic curve analysis, the area under the curve was increased to 0.831 for predicting PTC C4d+ when considering glomerulitis, DSAs, and HLA B eplet MM. The proportions of HLA I DSAs and PTC C4d+ in active antibody-mediated rejection were 12/17 and 15/17, respectively; the proportions of HLA class II DSAs and PTC C4d+ in chronic AMR were 8/12 and 7/12, respectively. Furthermore, the higher the PTC C4d+ score was, the more serious the urinary occult blood and proteinuria of recipients at the time of biopsy.@*CONCLUSIONS@#PTC C4d+ was mainly observed in AMR cases. DSAs, glomerulitis, and HLA B eplet MM are the independent risk factors for PTC C4d+.

Comparison of nighttime and daytime operation on outcomes of kidney transplant with deceased donors: a retrospective analysis / 中华医学杂志(英文版)

BACKGROUND@#Kidney transplant is always emergent operations and frequently need to be performed at nighttime to reduce cold ischemia time (CIT). Previous studies have revealed that fatigue and sleep deprivation can result in adverse consequences of medical procedures. This study aimed to evaluate whether nighttime operation has adverse impact on kidney transplant.@*METHODS@#A retrospective analysis of recipients accepted kidney transplant from deceased donors in one center from 2014 to 2016 was performed. Daytime transplant was defined as operation started after 8 AM or ended before 8 PM and nighttime operation was defined as operation ended after 8 PM or started before 8 AM. The incidences of complications such as delayed graft function, acute rejection, surgical complications and nosocomial infections were compared between 2 groups. Student's t-test was used to analyze continuous variables such as serum creatinine (Scr) at 1-year of post-transplant. The Chi-square test was used to analyze categorical variables. Differences in recipients and graft survival were analyzed using Kaplan-Meier methodology and log-rank tests.@*RESULTS@#Among the 443 recipients, 233 (52.6%) were classified into the daytime group and the others 210 (47.4%) were in the nighttime group. The 1-year survival rate of recipients was similar for the recipients in the daytime and nighttime groups (95.3% vs. 95.2%, P = 0.981). Although the 1-year graft survival rate in the nighttime group was slightly superior to that in the daytime group, the difference was not significant (92.4% vs. 88.4%, P = 0.164). Furthermore, Scr and incidence of complications were also not significantly different between the 2 groups.@*CONCLUSIONS@#Our results suggested that operation time of kidney transplant with short CIT has no significant impact on the outcome of kidney transplant. Nighttime operation of kidney transplant with short CIT could be postponed to the following day to alleviate the burden on medical staffs and avoid the potential risk.

Predictive Score Model for Delayed Graft Function Based on Hypothermic Machine Perfusion Variables in Kidney Transplantation / 中华医学杂志(英文版)

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 assessment.@*Methods@#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 model.@*Results@#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 quality of DCD kidneys, and it is potentially useful for physicians in making optimal decisions about the organs donated.

Beneficial Effect of Moderately Increasing Hypothermic Machine Perfusion Pressure on Donor after Cardiac Death Renal Transplantation / 中华医学杂志(英文版)

Background@#Vascular resistance and flow rate during hypothermic 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, 2015. HMP pressure was increased from 30 to 40 mmHg (1 mmHg = 0.133 kPa) 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 group. Among patients with DGF, kidney function recovery time and DGF duration were ameliorated in the IP group. Multivariate 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% CI: 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 DGF. Cox proportional hazards analysis showed that terminal resistance (hazard ratio: 2.06, 95% CI: 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.

Early Immunosuppressive Exposure of Enteric-Coated-Mycophenolate Sodium Plus Tacrolimus Associated with Acute Rejection in Expanded Criteria Donor Kidney Transplantation / 中华医学杂志(英文版)

<p><b>Background</b>Immunosuppressive agents are still inefficient in preventing biopsy-proven acute rejection (BPAR) after expanded criteria donor (ECD) kidney transplantation. The aim of this study was to investigate the relationships between early immunosuppressive exposure and the development of BPAR.</p><p><b>Methods</b>We performed a retrospective study of 58 recipients of ECD kidney transplantation treated with enteric-coated-mycophenolate sodium, tacrolimus (Tac), and prednisone. The levels of mycophenolic acid-area under the curve (MPA-AUC) and Tac Cwere measured at the 1 week and the 1 month posttransplant, respectively. The correlation was assessed by multivariate logistic regression.</p><p><b>Results</b>The occurrence rates of BPAR and antibody-mediated rejection were 24.1% and 10.3%, respectively. A low level of MPA-AUC at the 1 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-AUClevel was <30 mg·h·L at the 1 week (15.0% vs. 44.4%) or the Tac Cwas <4 ng/ml at the 1 month posttransplant (33.3% vs. 21.6%). Multivariable logistic regression analysis showed that the MPA-AUC at the 1 week (OR: 0.842, 95% CI: 0.784-0.903) and the Tac Cat the 1 month (OR: 0.904, 95% CI: 0.822-0.986) had significant inverse correlation with BPAR (P < 0.05).</p><p><b>Conclusions</b>Low-level exposure of MPA and Tac Cin the early weeks posttransplant reflects an increased acute rejection risk, which suggested that MPA-AUC <30 mg·h·L and Tac C <4 ng/ml should be avoided in the first few weeks after transplantation.</p>

Predictive Score Model for Delayed Graft Function Based on Easily Available Variables before Kidney Donation after Cardiac Death / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>How to evaluate the quality of donation after cardiac death (DCD) kidneys has become a critical problem in kidney transplantation in China. Hence, the aim of this study was to develop a simple donor risk score model to evaluate the quality of DCD kidneys before DCD.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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 four risk categories of increasing severity (scores 0-4, 5-9, 10-14, and 15-28).</p><p><b>CONCLUSIONS</b>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.</p>

Polyglycolic Acid Fibrous Scaffold Improving Endothelial Cell Coating and Vascularization of Islet / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Improving islet graft revascularization has become a crucial task for prolonging islet graft survival. Endothelial 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 islet. However, the traditional method of EC coating of islets has low efficiency in vitro. This 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 graft.</p><p><b>METHODS</b>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 angiogenesis.</p><p><b>RESULTS</b>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 culture. Stimulation 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 ECs-coating. For 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 (Tie-2) gene increased in PGA scaffold group than that in control group by real-time reverse transcription-polymerase chain reaction analysis.</p><p><b>CONCLUSIONS</b>These results demonstrate that the efficiency of EC coating of islets was successfully increased by culturing ECs on a PGA scaffold. This method enhances the function, survival, and vascularization of isolated islets in vitro and in vivo.</p>

Open surgical insertion of Tenkchoff straight catheter without guide wire / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To compare the clinical outcomes of open surgical peritoneal dialysis catheter (PDC) insertion with guide wire and the outcomes of PDC insertion without guide wire.</p><p><b>METHODS</b>Data of the patients receiving open surgical Tenkchoff straight catheter insertion in our department from January 2005 to January 2011 were retrospectively analyzed. The 117 patients in whom PDC insertion was conducted with the guidance of guide wire were enrolled into group A, and the 121 cases receiving PDC insertion without guide wire were enrolled into group B. The incidences of post-operative complications (catheter obstruction, catheter displacement, bloody dialysate, and dialysate leakage), catheter survival, and patient survival rates were compared between the 2 groups.</p><p><b>RESULTS</b>The baseline characteristics (gender, age, body mass index, prothrombin time, activated partial thromboplastin time, platelet count, serum creatinine, follow-up time, primary diseases, and outcomes) of the 2 groups were comparable (all P>0.05). In post-operative complications, only the incidence of early bloody dialysate showed significant difference, being 16.2% in group A and 7.4% in group B (P=0.04). Catheter and patient survival rates were not significantly different between the two groups. Overweight patients showed a higher incidence of catheter obstruction compared with normal weight patients [16.0% (4/25) vs.3.3% (7/213), P=0.02], but no differences in post-operative complications were found among overweight patients between the 2 groups.</p><p><b>CONCLUSIONS</b>Open surgical Tenkchoff straight catheter insertion without guide wire does not lead to higher risk of post-operative complications and catheter removal. It may be an alternative option when guide wire is not available.</p>

Preventive effect of poria cocos on acute rejection of renal transplantation in rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Poria cocos (Pcs) in preventing acute rejection of rats after renal transplantation and its mechanism.</p><p><b>METHODS</b>Rat orthotopic renal transplantation model was performed with Wistar rat as donor and SD rat as donee. All donees were divided into 4 groups, 10 in each group, before transplantation. They were treated respectively with normal saline 5 mL x kg(-1) x d(-1) (A), Pcs 25 mg x kg(-1) x d(-1) (B), Pcs 50 mg x kg(-1) x d(-1) (C) and ciclosporin A (CsA) 5 mg x kg(-1) x d(-1) (D) by intragastric administration. The renal allograft survival time (ST) was recorded, and the serum levels of creatinine (SCr), interleukin-2 (IL-2), gamma-interferon (gamma-IFN), CD4+, CD8+ lymphocytes percentage, CD4+/CD8+ ratio, as well as the pathologic changes were observed one week after transplantation.</p><p><b>RESULTS</b>ST of the renal graft in Groups C and D was significantly longer with pathologic change evidently less than those in Groups A and B (P<0.01), and the ST in Group C was shorter that in Group D (P<0.05). Changes of renal function and urine volume were identified to the pathological change of graft, the initiating time of renal dysfunction was later in Groups C and D than that in Groups A and B. Serum levels of IL-2, IFN-gamma and CD4+ percentage in Group C were significantly lower than those in Groups A and B, but higher than those in Group D respectively (P<0.05 or P<0.01), while CD8+ percentage in Group C was significantly lower than that in Group A (P<0.05), but insignificantly different to that in Groups B and D (P>0.05).</p><p><b>CONCLUSION</b>Pcs shows good dosage-dependent effect in suppressing acute rejection of renal transplantation, but the effect is inferior to that of CsA.</p>

Construction and identification of Ksp-cadherin-Gpx1-Klk1 expression vector / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a Gpx1 and klk1 recombinant vector containing the kidney-specific promoter Ksp-cadherin.</p><p><b>METHODS</b>Human Gpx1, Klk1 and Ksp-cadherin cDNAs were amplified with PCR and inserted in a stepwise manner into the expressive vector pIRES-EGFP to construct the recombinant vector Ksp-cadherin-Gpx1-Klk1. The constructed vector was verified with restriction enzyme digestion and sequence analysis.</p><p><b>RESULTS AND CONCLUSION</b>The recombinant expression vector Ksp-cadherin-Gpx1-Klk1 was constructed and identified successfully, which provides a potent tool for preparing transgenic animals to investigate gene therapy for ischemia-reperfusion injury in kidney transplantation.</p>

Application of kushenin on patients with chronic hepatitis C after renal transplantation / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of kushenin in treating patients with chronic hepatitis C after renal transplantation.</p><p><b>METHODS</b>Fifty-five patients were randomly assigned by lottery to the treatment group (29 cases) and control group (26 cases). The same immunosuppression therapy was given to all patients in both groups. Patients in the treatment group were treated with kushenin 0.6 g once a day, while those in the control group were treated with conventional liver protective agents such as vitamins. The treatment duration of both groups was 3 months. The incidences of serious hepatitis and acute rejection reaction, serum biochemistry parameters including indicators of liver and kidney functions, hepatic fibrosis index, and serum HCV-RNA were compared between the two groups.</p><p><b>RESULTS</b>(1) The incidence of serious hepatitis in the treatment group and the control group was 3.45% (1/29 cases) and 11.54% (3/26 cases), respectively, which was insignificantly different between the two groups (P=0.335). (2) The incidence of acute rejection in the treatment group was 6.90% (2/29 cases) and that in the control group was 7.69% (2/26 cases), showing insignificant difference (P=0.335). (3) The differences in serum alanine aminotransferase (ALT), direct bilirubin (DBIL), hyaluronic acid (HA), propeptide collagen type III (PC III), laminin (LN), collagen type IV (Col IV) levels between the two groups were insignificant before transplantation (P>0.05), while the above-mentioned parameters in the treatment group were significantly lower than those in the control group after transplantation (P<0.05). The difference in serum creatinine (SCr) and endogenous creatinine clearance rate (CCr) between the two groups was insignificant before and after transplantation (P>0.05). (4) The negative conversion rate of HCV-RNA in the treatment group was 31.03% (9/29 cases), significantly higher than the value of 11.54% (3/26 cases) in the control group after transplantation (P<0.05). (5) The levels of serum ALT and DBIL in patients with HCV-RNA converted to negative were significantly lower than those with still-positive HCV-RNA (P<0.05).</p><p><b>CONCLUSIONS</b>Kushenin has a certain effect on inhibiting the proliferation of HCV, protecting liver cells, and anti-liver fibrosis. On the other hand, it has no obvious influence on renal allograft function. Thus, the drug is clinically safe and effective for use in treating patients with chronic hepatitis C after renal transplantation.</p>

Directional differentiation of murine CD117+ hemopoietic stem cells into immature dendritic cells and their identification / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a stable method for obtaining large quantity of highly purified immature dendritic cells (imDCs) in vitro, and identify the morphology, function and surface markers of the cells.</p><p><b>METHODS</b>CD117(+) hemopoietic stem cells (HSCs) were isolated and purified from the bone marrow of healthy C57 mice by magnetic affinity cell sorting. After cell expansion by treatment with stem cell factor (SCF) and interleukin-3 (IL-3), the HSCs were induced for directional differentiation into imDCs by treatment with GM-CSF, IL-4 and IL-10. The imDCs obtained were identified by morphological and functional observation under inverted microscope, scanning electron microscope and transmission electron microscope, followed by detection of the expressions of the surface markers using flow cytometry.</p><p><b>RESULTS</b>After 3, 5 and 7 days of culture in the presence of SCF+IL-3, the cells were expanded by 10.34-/+1.43, 22.65-/+2.71 and 54.39-/+3.08 folds, respectively. The HSCs were successfully induced to differentiate into imDCs with phagocytotic activity. The dendrites of the imDCs were short small, and appearing spinous. The expressions of surface markers were detected from the cells showing the phenotype of CD11c(+), I-A/I-E(low), CD40(-), CD80(-), CD86(-).</p><p><b>CONCLUSION</b>The method described allows steadily acquisition of large quanty of highly purified imDCs and of their effective identification in vitro.</p>

Impacts of kidney transplantation on erectile function and its contributing factors evaluation / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the impacts of kidney transplantation on erectile function and analyse its contributing factors.</p><p><b>METHODS</b>In order to evaluate the severity of erectile dysfunction (ED), a total of 250 married male kidney transplant recipients (KTR) with functioning graft were assessed with the International Index of Erectile Function (IIEF) questionnaire. Data of clinical characteristics, medical and sexual history and laboratory examination were collected. Univariate and multivariate logistic regression analyses were carried out to determine which have independent impacts on erectile function.</p><p><b>RESULTS</b>The investigation was accomplished in 84.8% of the KTRs. There was no significant difference in ED incidence before and after renal transplantation (53.8% vs. 44.3%, P > 0.05). According to the IIEF score, erectile function improved in 43.9% of the KTRs, remained unchanged in 42.9%, and deteriorated in 13.2%, as compared with pre-transplantation. Logistic regression analysis showed that significant and independent influencing factors in erectile function were age, hemoglobin level, presence of DM and/or peripheral neuropathy and iterative transplantations, and their relative risks were 3.01, 2.01, 3.15, 3.89 and 2.67, respectively.</p><p><b>CONCLUSION</b>ED is highly prevalent among KTRs and its pathogenesis is multifactorial. Age, presence of DM and/or peripheral neuropathy, hemoglobin level and iterative transplantations were chief contributing factors in erectile function.</p>

Co-expression of sCD40LIg and CTLA4Ig mediated by adenovirus prolonged mouse skin allograft survival / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

<p><b>OBJECTIVE</b>To investigate the role of simultaneous blockade of CD40/CD40L and B7/CD28 pathways in the immune tolerance via co-expression of sCD40LIg and CTLA4Ig mediated by replication-defective adenovirus.</p><p><b>METHODS</b>Ad-sCD40LIg-IRES(2)-CTLA4Ig, replication-defective adenovirus co-expressing sCD40LIg and CTLA4Ig, was constructed and identified. The co-expression of sCD40LIg and CTLA4Ig was evaluated with confocal laser scanning microscope and Western blotting. Skin transplantations of C57BL/6 to BALB/c mice were performed. PBS, Ad-Shuttle-CMV and Ad-sCD40LIg-IRES(2)-CTLA4Ig were administered. Skin graft survival was monitored and the mRNA expression of both genes was evaluated in the skin allografts.</p><p><b>RESULTS</b>Ad-sCD40LIg-IRES(2)-CTLA4Ig was constructed successfully and identified. The co-expression of sCD40LIg and CTLA4Ig was identified with confocal laser scanning microscopy and Western blotting. Compared to the skin graft mean survival time (MST) of non-treated group ((5.75+/-0.71) d) or Ad-Shuttle-CMV-treated group ((5.50+/-0.53) d), the skin graft MST was dramatically prolonged in the Ad-sCD40LIg-IRES(2)-CTLA4Ig-treated group ((16.38+/-1.19) d, P<0.001). The mRNA expression of both genes was detected.</p><p><b>CONCLUSION</b>Ad-sCD40LIg-IRES(2)-CTLA4Ig, a replication-defective adenovirus carrying genes encoding sCD40LIg and CTLA4Ig, was constructed. Simultaneous blockade of CD40/CD40L and B7/CD28 costimulatory pathway mediated by replication-defective adenovirus significantly prolonged skin allograft survival in mice.</p>

Cryopreservation of microencapsulated rat islets / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of alginate-polylysine-alginate (APA) microcapsules in protecting rat islet cells in cryopreservation.</p><p><b>METHOD</b>Purified rat islet cells microencapsulated with APA and free islet cells were cryopreserved for one month and then thawed for culture in RPMI 1640 overnight. The morphology of the cells was observed and their function assessed by stimulated insulin release test.</p><p><b>RESULT</b>APA microcapsulation protected the fragile islets from freezing damage by increasing the recovery rate of the cells from 68.6%+/-2.9% to 94.7%+/-1.4% (P<0.05). After incubation with high glucose (16.7 mmol/L) solution, the insulin release from the encapsulated cells after cryopreservation significantly increased in comparison with that of the nonencapsulated cells (22.6+/-1.8 mU/L vs 11.7+/-1.5 mU/L, P<0.05). In high glucose solution containing theophylline, the calculated stimulation index of the encapsulated cells was about 3 times that of the nonencapsulated cells.</p><p><b>CONCLUSION</b>APA microencapsulation may significantly increase the post-thaw recovery and improve the function for cryopreserved rat islets.</p>

Recombinant adenovirus-mediated human cytosolic glutathione peroxidase gene transfection protects vascular endothelial cells from oxidative damage / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the protective effect of recombinant adenovirus-mediated human cytosolic glutathione peroxidase (hCGPx) gene transfection on vascular endothelial cells ECV304 from oxidative damage.</p><p><b>METHODS</b>pGEM-T Easy Vector containing hCGPx cDNA and recombinant adenovirus shuttle plasmid pACCMV-pLpA were used to construct the shuttle plasmid pACCMV-hCGPx for cotransfection of 293 cells with pJM17, thereby to obtain the recombinant adenovirus AdCMV-hCGPx. Cultured ECV304 cells were transfected with AdCMV-hCGPx for 24, 48 and 72 h, respectively, with the cells transfected with the empty vector serving as control, and hCGPx gene expression was then examined in the transfected cells. The transfected cell viability and apoptotic cell ratio were evaluated after treatment of the cells with H(2)O(2).</p><p><b>RESULTS</b>The expression ratio of hCGPx gene was significantly higher in the AdCMV-hCGPx-transfected cells than in those with empty vector transfection (P<0.01). The hCGPx gene-transfected cells showed significantly higher viability and significantly lower apoptotic ratio than the control cells following challenge with H(2)O(2)-induced oxidative damage.</p><p><b>CONCLUSION</b>hCGPx gene transfer mediated by recombinant adenovirus protects the vascular endothelial cells from oxidative damage in vitro, possibly due to the antioxidative and apoptosis-inhibiting effect of hCGPx.</p>

Isolation and culture of adult Sertoli cells and their effects on the function of co-cultured allogeneic islets in vitro / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Globally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered 'nurse cells' in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro.</p><p><b>METHODS</b>Adult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets' function when they were co-cultured for 21 days in vitro.</p><p><b>RESULTS</b>In cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P > 0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P < 0.01).</p><p><b>CONCLUSIONS</b>A method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets' function when they were co-cultured in vitro. They could be a helper cell in islet transplantation.</p>

Application of Danshen injection on early stage of renal transplantation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Danshen injection (DSI) on early stage of renal transplantation.</p><p><b>METHODS</b>One hundred and twelve patients in early stage after renal transplantation were allocated in the treated group, they were treated by conventional treatment with DSI 60 ml given additionally once a day for 10 days. And 109 patients who received conventional treatment alone after renal transplantation at the corresponding period were allocated in the control group. Indexes in the two groups, including volume of urine, serum creatinine (SCr), endogenous creatinine clearance rate, incidence of delayed graft function and acute rejection reaction, blood viscosity (BV), platelet aggregation rate (PAR) as well as the blood flow resistance in graft measured by color Doppler ultrasonography.</p><p><b>RESULTS</b>The urinary volume and endogenous creatinine clearance rate in the treated group were significantly higher, but levels of SCr, incidence of renal function recovery retardation, BV, PAR and blood flow resistance in graft were significantly lower than those in the control group (P < 0.05). The difference of incidence of acute rejection reaction between the two groups was insignificant (P > 0.05).</p><p><b>CONCLUSION</b>DSI can improve blood microcirculation, decrease the incidence of renal function recovery retardation, these effects are helpful for recovery of renal function after renal transplantation.</p>

Efficacy of Hushen Gujing (HSGJ) in preventing chronic allograft nephropathy in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the effect of HSGJ on chronic allograft nephropathy (CAN) using standard rat model of CAN.</p><p><b>METHOD</b>Renal transplantation was performed with Fisher rats as donors and Lewis rats as recipients. All the recipients were randomly divided into control group and medication groups (high and low dosage of HSGJ, fed every other day). After 16 weeks of treatment, renal function and the histological alteration of CAN were measured. The expression of the TGFbeta1 mRNA in the allograft was evaluated by real-time PCR.</p><p><b>RESULT</b>The content of 24 h urine protein and the level of serum creatinine in the medication groups were significantly decreased (P < 0.01) as compared with control group, whereas the creatinine clearance was increased (P < 0.01). The degree of glomerular sclerosis and the Banff score of medication groups were lower than the control group respectively (P < 0.01), in consistent with decreased expression of the TGF 1mRNA.</p><p><b>CONCLUSION</b>HSGJ can prevent the chronic allograft nephropathy and the mechanism may be related with its influence on the expression of the TGFbeta1.</p>