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Transplantation of en-bloc kidneys and single kidney from pediatric donors to pediatric recipients: 11 cases report

Liang ZHU ; Wenyu ZHAO ; Li ZENG ; Lei ZHANG ; Fanyuan ZHU ; Youhua ZHU

Chinese Journal of Organ Transplantation.2014;35(5):287-289. doi:10.3760/cma.j.issn.0254-1785.2014.05.008

Objective To investigate the therapeutic principle of en-bloc kidney and single kidney transplantation from pediatric donors to pediatric recipients.Method A retrospective analysis of 11 pediatric kidney transplants into pediatric recipients was performed.The age of donors and recipients was 33 days to 48 months,and (9.1 ± 3.4) years (4.6 14.3 years) respectively.Result During the follow-up period of 1 to 22 months,the patient survival rate was 100％.Complications included delayed graft function in 1 case (managed by peritoneal dialysis),urine leak in 2 cases (treated by reoperation),hydronephrosis in 2 cases (treated by extracting ureteral catheter) and vascular thrombosis in 1 case.Due to thrombosis,one graft was lost.Of the remaining 10 recipients,all had excellent long-term function.At the last follow-up,their serum creatinine levels were 65.5 ±13.6 μmol/L (49-83μmol/L),and transplanted renal ultrasound examination showed no abnormality.Conclusion Kidney grafts from pediatric donors can be successfully transplanted to pediatric recipients,but the therapeutic principle is different from that in adult kidney transplantation.

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The analysis of anti-major histocompatibility complex class Ⅰ-related chain A(MICA) specific antibodies testing results in multicenter and its clinical applications

Xiaoni YUAN ; Jun HE ; Jianquan HOU ; Xiaojing BAO ; Chao XU ; Yang LI ; Jingcheng MIAO

Chinese Journal of Organ Transplantation.2014;35(5):290-294. doi:10.3760/cma.j.issn.0254-1785.2014.05.009

Objective To research the consistency of testing results with three different antimajor histocompatibility complex class Ⅰ-related chain A(MICA) specific antibody reagents in order to evaluate their clinical application's value.Method An collaborative study of 18 laboratories was undertaken at the 16th International HLA and Irnmunogenetics Workshop.Total of 16 sera(4 batchs)were tested for anti-MICA antibodies by Luminex method with three different reagents (Kit-A,-B and -C).Result Anti-MICA antibodies were found in 15 sera,except one sera(no.S04) ; No.S10 sera showed positive results in all the laboratories.The anti-MICA antibodies were divided into MICA-G1 group (MICA01,02,07,12,17 and 18) and MICA-G2 group (MICA 04,06,08/27,09 and 19).MICA-G1 group specific antibodies were detected in 5 sera with Kit-A and-B reagent; but there were false-positive results of anti-MICA08/27 and MICA19 antibodies in this 5 sera with Kit-C.MICA-G2group specific antibodies can be detected in other 5 sera with Kit-A and-B,But the MICA specific antibodies testing gave different results with Kit-A,-B and-C in all the last 5 sera samples.Testing of MICA08/27 showed highest consistency results (86.67％,13/15) with Kit-A,-B and-C; and testing of MICA19 showed lowest consistency results (40％,6/15) with this 3 reagents.There were 80％ consistency results of anti-MICA specific antibodies in 13 sera with Kit-B.Conclusion There are the same effect to judgment positive or negative result for anti-MICA antibodies with 3 different reagents,but the results of anti-MICA specific antibodies are not the same.Therefore,it's better to use two or more reagents to test anti-MICA specific antibodies,or choose reagent with wide detection range.

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Establishment of orthotopic liver transplantation model in pigs without veno-venous bypass

Junfeng HAN ; Wei GAO ; Lei LIU ; Yuan SHI ; Mei DING ; Zhongyang SHEN

Chinese Journal of Organ Transplantation.2014;35(5):310-312. doi:10.3760/cma.j.issn.0254-1785.2014.05.013

Objective To summarize the experience of orthotopic liver transplantation in pigs without veno-venous bypass.Method In general,Bama miniature pigs were used as both the donors and the recipients.Suprahepatic inferior vena cava and portal vein anastomosis was performed with running prolene sutures.After completion of the portal vein anastomosis,the graft was reperfused.Infrahepatic inferior vena cava anastomosis and hepatic artery anastomosis were performed in a similar fashion.Finally,the common bile duct was reconstructed.Result For all of the transplant procedures,the average cold ischemic time was 356.3 ± 66.4 min and anhepatic time 22.5 ± 2.6 min,and the average operative time was 185.7 ± 24.8 min.During the anhepatic phase,the central venous pressure (CVP) and the mean arterial pressure (MAP) were significantly lower than those at baseline (P＜ 0.05).Heart rate (HR),on the other hand,was increased significantly during the anhepatic phase (P＜0.05).By the time the portal vein and the hepatic artery were reperfused,and CVP and MAP were gradually elevated,and HR gradually reduced.All receptors were successfully extubated and awake after surgeries.On the third postoperative day they began to eat.All receptors survived during the intraoperative period,and the survival rate was 93.8％ (15/16) on the fifth postoperative day.One receptor was died on the third postoperative day due to abdominal infection.Conclusion This model has satisfactory stability and reproducibility.Without using any vasoactive substances,to maintain the MAP beyond 50 mmHg in the anhepatic phase and the short anhepatic time are important to perform successful liver transplantation.

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Dynamic monitoring of donor specific antibodies in living-relative renal transplantation and early intervention

Yu CHEN ; Jiangyan LI ; Xiaoyan LOU ; Hao WANG ; Lei ZHANG ; Youhua ZHU

Chinese Journal of Organ Transplantation.2014;35(5):267-269. doi:10.3760/cma.j.issn.0254-1785.2014.05.003

Objective To analyze the clinical application of HLA donor specific antibodies (DSAs) detected by Luminex single antigen beads,and to discuss the impact of early intervention on renal function.Method In 64 cases of living-relative renal transplantation,DSA was detected using a Luminex single antigen assay before and after transplantation.The positive recipients were given large doses of intravenous irnmunoglobulin (IVIG) and increased doses of mycophenolate mofetil (MMF).The relationship between DSA and renal function was analyzed.Result DSA was negative in all recipients before transplantation.Ten cases of DSA positive recipients were found in HLA mismatch after transplantation.After the intervention,two cases of DSA positive recipients became negative,immunofluorescence intensity was decreased by more than 50％ in 6 cases,and no significant reduction was found in the other two cases.Antibody-mediated rejection (AMR) occurred in two cases of intervention ineffective recipients after 3 to 6 months and the renal function was impaired.Conclusion Dynamic monitoring of DSA using Luminex single antigen beads may timely predict changes of renal function.Early application of large doses of IVIG and increasing doses of MMF can reduce the incidence of AMR.

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The clinical pattern and treatment of 30 early renal allograft recipients with interstitial pneumonia

Xiaoxiao WANG ; Song CHEN ; Jun YU ; Lan ZHU ; Zhengbin LIN ; Gang CHEN

Chinese Journal of Organ Transplantation.2014;35(5):282-286. doi:10.3760/cma.j.issn.0254-1785.2014.05.007

Objective To investigate the clinical pattern,therapeutic principle and influencing factors of interstitial pneumonia in renal allograft recipients.Method The general information,clinical manifestation,treatment and outcomes of 30 recipients with interstitial pneumonia after renal transplantation from Nov.2006 to Dec.2013 were analyzed retrospectively.Result Twenty-nine of 30 patients developed interstitial pneumonia between 2 to 6 months post-transplant.The total course of the pneumonia lasted for 34.9 ± 7.5 days on average.Chest CT scans were used to monitor severity of interstitial pneumonia each week.The mean duration between the onset to the fastigium of pneumonitis was approximately 14.8 ± 1.9 days.The mean duration of the fastigium lasted for 7.3 ±3.6 days,after that the patients usually started to recover.Deteriorated chest CT scan findings and long terms of the fastigium usually indicated poor outcomes.The mean duration of the recovery period was 13.1 ± 3.7 days.After adjusted administration of methylprednisolone,antibiotics,antifungal agents,nutritional support as well as immunosuppressive drugs,23 patients with mild and moderate pneumonia by the chest CT scans were cured and discharged.However,4 of the 7 patients with severe pneumonia by the chest CT scans died.Conclusion The progression of interstitial pneumonia after renal transplantation is characterized by a more consistent regularity.After adjusted administration of methylprednisolone,antibiotics,antifungal agents,nutritional support as well as immunosuppressive drugs,renal allograft recipients with interstitial pneumonia could obtain a good therapeutic effect without over-treatment.

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Correlation between miR-494 and TH17 cell differentiation in murine cardiac transplant rejection

Shu HAN ; Youhua ZHU ; Liming WANG ; Li ZENG ; Shangxi FU ; Xueyang ZHENG

Chinese Journal of Organ Transplantation.2014;35(5):295-299. doi:10.3760/cma.j.issn.0254-1785.2014.05.010

Objective To investigate correlation between microRNA (miR-494) and TH 17 cell differentiation in murine cervical heterotopic cardiac transplant model.Method The heterotopic cardiac transplant models of Balb/c→C57BL/6 mice were established as experimental group,and those of C57BL/6→C57BL/6 mice as control group.Real time-polymerase chain reaction(PCR) was used to detect miR-494 and interleukin(IL)-17A mRNA expression in the grafts.CD4+ T cells,CD8+ T cells and CD45+ myeloid cells were isolated from the grafts,and miR-494 and IL-17 mRNA expression was detected.In vitro,lymphocytes in the spleen from C57BL/6 mice were harvested,and CD4+ T cells were isolated with MACS and then stimulated to TH 1,TH 2,TH 17,Treg subset cells.The expression of IL-17A mRNA and miR-494 in different T subsets was examined by Reverse transcription-polymerase chain reaction(RT-PCR).Result Two grafts from each study group were harvested on the 7th day post-transplantation.In experimental group,the IL-17A mRNA expression was increased,while the expression of miR-494 was decreased as compared with control group with the difference being significant between two groups.The expression of IL-17A rnRNA in CD4+ T cells of the grafts was significantly increased,while that expression of miR-494 was decreased.In vitro,the expression of miR-494 in TH 17 cells was significantly lower than that in TH 1,TH 2 and Treg cells.Conclusion miR-494 is related closely to TH 17 cells differentiation in the transplant rejection,which may play a role in transplant rejection through regulating TH 17 cells.

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Measurement of ABO blood group antibody levels using renal cortical tubular epithelial cells of cynomolgus monkey

Lu WANG ; Junxiang WANG ; Xiaoxiao WANG ; Song CHEN ; Gang CHEN

Chinese Journal of Organ Transplantation.2014;35(5):300-305. doi:10.3760/cma.j.issn.0254-1785.2014.05.011

Objective To investigate the possibility of measuring ABO blood group antibody levels using renal cortical tubular epithelial cells (RCTECs) of cynomolgus monkey.Method The primary RCTECs were isolated from cynomolgus monkey kidneys and identified by Western blotting and flow cytometry (FACS).FACS was applied to detect the expression of ABO histo-blood group antigens on RCTECs,the binding of blood group antibodies in human serum to RCTECs,and to compare the difference of measuring ABO blood group antibody levels between using human erythrocytes and RCTECs as target cells.Result The majority of cells derived from the kidney cortex were renal tubular epithelial cells.39.90％-73.80％ of RCTECs were found to express the ABO histoblood group antigens with the capability to bind the human blood group antibodies specifically.The expression level of ABO histo-blood group antigen on RCTECs was not influenced by long-term cryopreservation,and the percentage of B antigen positive cells was 39.90％ before frozen storage and 39.65％ after a 10-month cryopreservation respectively.The normal serum and allergic serum presensitized to synthetic A antigen-keyhole limpet hemocyanin (KLH-A) of cynomolgus monkey were used to compare the abilities of RCTECs and human erythrocytes to detect antibody titers.No agglutination was observed in RCTECs test group.The mean fluorescence intensity (MFI) values of antibody were highest when the dilution fold was lowest (1 ∶ 16) and gradually decreased with increased serum dilution in both serum groups.MFI fell towards baseline value at 1 ∶ 128 dilution in normal serum group while at 1∶8 192 dilution in allergic serum group.Between 1 ∶ 16 and 1∶8 192 dilutions,MFIs of normal serum group were all lower than those of allergic serum.In human red cells test group,obvious agglutination appeared at high concentrations of antibodies,and MFIs reached the peak at 1∶64 dilution in normal serum group and at 1∶32 dilution in allergic serum group,but fluctuated irregularly thereafter.Between 1∶64 and 1∶512 dilutions,MFIs of normal serum group were all higher than those of allergic serum.Data above showed that MFI values in RCTECs group could reflect the levels of blood group antibodies more exactly.Conclusion The RCTECs of cynomolgus monkey express ABO tissue-blood group antigens and would be more suitable to be used to measure ABO blood group antibody levels by FACS.

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Conspecific mesenchymal stem cells infusion for immune thrombocytopenia and the impact on CD4+CD25highCD127low regulatory T cells

Min XU ; Guoqiang LIU ; Xia ZHAO ; Guang LU ; Huifang DING ; Liang WANG ; Caifeng SUN

Chinese Journal of Organ Transplantation.2014;35(5):306-309. doi:10.3760/cma.j.issn.0254-1785.2014.05.012

Objective To evaluate the effect of the conspecific bone marrow mesenchymal stem cells (MSCs) infusion on the platelets count and the ratio of CD4+ CD25highCD127low regulatory T (Treg) cells in mice with immune thrombocytopenia and the mechanisms.Method ITP mice models were induced by daily intraperitoneal injection of 200 μL phosphate buffer solution [containing 2 μg rat anti-platelet membrane CD41 antibody (MWReg30)] into female Balb/c mice.MSCs were got from male mice.Then different number of MSCs was injected into ITP mice through the tail veins.After 5,7 and 14 days,the number of blood platelets was counted and the ratio of Treg cells was detected by flow cytometry,and compared with those in the healthy mice.Result Twenty-four h after injection of CD41 antibody,platelet counts were reduced sharply to the lowest point,which was about a quarter of the normal level.Then ITP mice models were induced successfully.Platelet counts were increased after the injection of MSCs.On 7th day after injection of MSCs,the platelet counts were significantly higher than those in control mice,and the greater the degree of injection dosage,the greater the elevated platelets (P＜0.05 for all).The ratio of Treg cells in ITP mice models was significantly lower than in the normal mice.The ratio of peripheral blood Treg cells in ITP mice was increased after injection of MSCs and the higher the dose,the greater the effect (P＜0.05 for all) but did not reach the normal level.Conclusion The conspecific bone marrow MSCs infusion can increase the platelet counts in mice with ITP,which may be related to the increase of CD4+ CD25highCD127low Treg cells.

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The prognostic value of combined analysis of KLF4 expression and the Milan criteria in hepatocellular carcinoma patients after orthotopic liver transplantation

Hongcheng SUN ; Zhihai PENG

Chinese Journal of Organ Transplantation.2014;35(6):333-336. doi:10.3760/cma j.issn.0254-1785.2014.06.004

Objective To evaluate the prognostic value of combined analysis of the biomarker Krüppel-like factor 4 (KLF4) and the Milan criteria in hepatocellular carcinoma (HCC) patients treated by orthotopic liver transplantation (OLT).Method The clinicopathological data and outcome of the recruited 105 HCC patients undergoing OLT from October 2001 to April 2009 were retrospectively analyzed.KLF4 expression in HCC and paired non-tumor tissue was detected by immonohistochemistry and confirmed by Western blotting analysis.Five-year overall survival (OS) and recurrence-free survival (RFS) rate and survival curves of the grouped patients were calculated and plotted by Kaplan-Meier method.Result The level of KLF4 expression was lower in HCC than that in paired non-tumor tissue (P＜0.05).KLF4 expression was significantly lower in poorly differentiated HCC than that in well-moderately differentiated HCC (P =0.008).Loss of KLF4 was an independent risk factor for predicting tumor recurrence and survival of HCC after OLT (HR =0.459 and 5.42,respectively,P＜0.001).The level of KLF4 expression could not differentiate the OS and RFS rate in the patients with tumors meeting the Milan criteria,whereas the OS and RFS rate in the patients with tumors exceeding the Milan criteria differentiated according to KLF4 expression.The patients with tumors beyond the Milan criteria and exhibiting moderate to high KLF4 expression had unexpectedly favorable 5-year OS (91.7％) and RFS (70.5％) rate.Conclusion KLF4 is a useful biomarker for prognostication of HCC patients undergoing OLT.Integrated use of KLF4 biomarker and the Milan criteria improves accurate prediction of survival and tumor recurrence for HCC patients after OLT.

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Experience of donation after citizens death for lung transplant: report of 19 cases

Haiying XU ; Wenjun MAO ; Jingyu CHEN ; Xiaoling LIU ; Wenjuan ZHOU

Chinese Journal of Organ Transplantation.2014;35(7):396-400. doi:10.3760/cma.j.issn.0254-1785.2014.07.004

Objective To summarize the experience of harvesting and using the lungs from donation after citizens death.Method From November 2007 to December 2013,19 cases of potential donation after brain death (DBD) and donation after cardiac death (DCD) were evaluated,including 9 cases of DCD and 10 cases of DBD.All the patients received the tests of sputum culture,bedside bronchoscopes,chest X rays,and blood gas analysis.After clear evaluation,3 cases of DCD and 3 cases of DBD were discharged from the group for bilateral inflammatory infiltration and poor oxygenation index,and the rest one case of DCD was precluded due to long warm ischemic time (＞60 min).The donor lungs from remaining 12 cases were harvested successfully after the declaration of brain death or cardiac death.The donors suitable for the transplant procedure were transported to our transplant center.Result Twelve lung transplants were performed successfully,including 10 cases of bilateral lung transplantation and 2 cases of right single lung transplantation.Two patients was complicated with severe infection and died of sepsis postoperatively,and the remaining 10 patients all recovered uneventfully with dramatic improvement of pulmonary function.During the follow-up period,all the patients lived an active life style with high quality of life.The mean survival time was 34.7 months (4-60 months).Conclusion Lung transplantation using DCD and DBD can be successfully performed after adequate preoperative evaluation of donor lung and abundant preparation for donor harvesting.

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