器官移植

Abstract:
Objective To investigate the effect of micro ribonucleic acid (miRNA)-155 on two subtypes of regulatory T cell (Treg): induced Treg (iTreg) and natural Treg (nTreg). Methods Naïve T cells and nTreg were isolated from peripheral blood mononuclear cell(PBMC) of healthy donors by magnetic cell sorting. Cells were divided into 3 groups during culture, including control group(naïve T cells were cultured with the presence of interleukin-2), iTreg group(naïve T cells were cultured with the presence of interleukin-2 and transforming growth factor-β) and nTreg group(nTreg cells was cultured with interleukin-2). Each group was divided into 3 subgroups(none, scramble or miRNA-155 antagomir subgroup, 3 wells in each subgroup). Expression level of miRNA-155 gene of none subgroup in 3 groups was detected by low density chip analysis method. The levels of surface marker CD25, Foxp3, CD127 of each subgroup in 3 groups were detected by flow cytometry. The percentage of CD4⁺ CD25⁺Foxp3⁺SOCS1⁺ Treg and suppressive function of Treg of each subgroup in 3 groups were also detected by flow cytometry. Results Compared with control group and iTreg group, the expression level of miRNA-155 was significantly lower and SOCS1 was significantly higher in nTreg group(all in P<0.05). After the addition of miRNA-155 antagomir, no significant change was observed in the important surface markers of Treg like Foxp3, CD25, CD127. Compared with control group and iTreg group, the expression of SOCS1 in nTreg group increased significantly (both in P<0.05). The expression level of miRNA-155 of none subgroup in iTreg group was lower. The expression of SOCS1 increased after the miRNA-155 was inhibited by antagomir(miRNA-155 antagomir subgroup). In iTreg group, the suppressive function of Treg in miRNA-155 antagomir subgroup was higher than that in none subgroup at the ratio of 1:8, 1:16 and 1:32(all in P<0.05). Conclusion Antagonism of miRNA-155 in vitro has no significant effect on the suppression function of nTreg, but can increase the SOCS1 expression level and suppression in vitro of iTreg.

Experimental study on the Caspase-3 siRNA decreases apoptosis in stable cell lines of bone marrow mesenchymal stem cells

Liu Libao, Huang Lei, Zhang Junhang, Liao Hongying, Hu Ailing, Xu Shilei, Hua Ping

2014, 5(5): 283-288. doi: 10.3969/j.issn.1674-7445.2014.05.005

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Abstract:
Objective To explore a method of using Caspase-3 small interference ribonucleic acid (siRNA) in vitro to decrease the apoptosis in stable cell lines of bone marrow mesenchymal stem cell (MSC) in order to establish MSC cell lines with stable expression. Methods Virus particles containing Caspase-3 siRNA were generated in 293FT cells by retroviral packaging system, then were transfected into MSC of rats. And the transfected cells were screened and cultured to get the stable MSC lines. According to the characteristics of retrovirus, the stable expression of the cell lines was identified by Western blot and real-time fluorescent quantitative PCR (RT-QPCR). The apoptosis of stable cell lines and normal MSC were detected by terminal dexynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) and their differences were compared. Results Compared with the normal cells, the caspase-3 protein expression and mRNA content of stable MSC lines were significantly reduced. In the same condition in vitro, the apoptosis quantity of stable MSC lines significantly decreased compared with the normal cells. Conclusion Stable cell lines of MSC with stable expression of Caspase-3 siRNA can be obtained by retroviral packaging system. The apoptosis quantity of stable MSC Lines significantly decreased compared with the normal cells.

Comparison on liver transplantation rejection among various strains combination of rats

Ye Zhijian, Yang Zhongmeng, Wang Xiaomin

2014, 5(5): 289-293. doi: 10.3969/j.issn.1674-7445.2014.05.006

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Abstract:
Objective To compare the rejection of liver transplantation in different strains combination of rats. Methods Four transplantation models of various strain combinations were established including the control group SD (donor) to SD (recipient) (group A) and the experimental groups Wistar to SD(group B), Lewis to BN(group C) and DA to Lewis(group D). Orthotopic liver transplantation model of rats were constructed by Kamada's two cuff technique. And serum hepatic functions in 4 groups of recipient rats at 10^th day after liver transplantation were compared, which was including alanine aminotransferase (ALT), total bilirubin (TB) and albumin (Alb). Meanwhile the serum levels of interleukin(IL)-2 and IL-10, rejection activity index(RAI) at 10^th day after liver transplantation and the average survival time of recipient rats were investigated. Results Compared with group A and B, the serum levels of ALT and TB of group C and D increased obviously, while Alb decreased significantly. Compared with group C, TB of group D increased obviously, while Alb decreased remarkably. All of the differences were statistically significant (all in P<0.05). Compared with group A, the serum levels of IL-2 and IL-10 in group B, C and D all elevated significantly, and IL-2/IL-10 in group C and D increased remarkably too (all in P<0.05). Compared with group B, IL-2 in group C and D increased significantly, and IL-2/IL-10 in group D also elevated significantly. Compared with group C, both IL-2 and IL-2/IL-10 in group D increased significantly. All the differences were statistically significant (all in P<0.05). Hepatic histopathological examination showed that obvious rejection did not occur in the recipient rats of group A, which rejection activity index (RAI) was scored 1.8±0.7. The RAI score of Group B,C and D were 3.1±1.3, 6.9±0.8 and 8.8±0.5, which belonged to mild or vague rejection, moderate to severe rejection and severe rejection respectively. All the RAI scores comparison among and between each group had statistical differences (all in P<0.05). The average survival time of recipient rats in group A, B, C and D were (119.3±1.9) d, (116.9±8.3) d, (53.4±6.1) d and (12.1±2.4) d respectively. There was no statistically significant difference between group A and B, while the comparison between any other group was statistically significant (all in P<0.05). Conclusion In four different rats strains combination, the rejection is most serious in the model of DA (donor) to Lewis (recipient), moderate in Lewis to BN, mildest and near to tolerance in Wistar to SD.

Relationship between the expression of SDF-1, ICAM-1 and renal tubular necrosis score in rats with renal ischemic reperfusion injury

Ma Shuaijun, Zhang Geng, Cao Zhiqiang, Liu Kepu, Li Zhibin, Ruan Dongli, Yang Xiaojian, Yuan Jianlin

2014, 5(5): 294-298. doi: 10.3969/j.issn.1674-7445.2014.05.007

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Abstract:
Objective To investigate the relationship between the expression of stromal cell derived factor (SDF)-1, intercellular adhesion molecule (ICAM)-1 and renal tubular necrosis score in rats with renal ischemic reperfusion injury (IRI). Methods Sixty Sprague-Dawley (SD) rats were randomly divided into operation group and sham operation group with 30 rats in each group. Then each group was divided into 6 subgroups (1 h,6 h,12 h,24 h, 48 h or 72 h) according to different time of measurement after operation with 5 rats in each subgroup. Renal IRI mode1s were built in rats of operation group. The bilateral renal arteries were dissected in rats of sham operation group and then the incision was sutured. The renal function, renal tubular necrosis score and the variation of SDF-1, ICAM-1 expression in renal tissues at different time points were measured. The relationship between the expression of SDF-1, ICAM-1 in renal tissues and renal tubular necrosis score in rats of operation group was analyzed by Pearson correlation analysis. Results The levels of urea nitrogen (BUN) and serum creatinine (Scr) after operation in each subgroup of operation group were significantly higher than those before operation and those in the corresponding subgroups of sham operation group(all in P<0.05). They increased significantly 12 h after operation and reached the peak at 48 h after operation. The renal tubular necrosis score in operation group increased gradually over time (all in P<0.05). The highest renal tubular necrosis score was in the 48 h operation subgroup (P<0.05). Compared with those in 1 h operation subgroup, the expression of SDF-1, ICAM-1 in rats' renal tissues of 6 h operation subgroup began to increase significantly, and they reached the peak at 48 h after operation and began to drop down at 72 h after operation. The expression of SDF-1, ICAM-1 in rats renal tissues in operation group were positively correlated with the levels of BUN, Scr and renal tubular necrosis score at different time points after operation (r=0.614, 0.662, 0.751;0.640, 0.703, 0.785;P<0.05). Conclusion When rat's renal tissue develops IRI, the expression of SDF-1, ICAM-1, BUN, Scr and renal tubular necrosis score increased.The expression of SDF-1, ICAM-1 are positively correlated with BUN, Scr and renal tubular necrosis score. The increased expression of SDF-1, ICAM-1 can serve as an indicator of the severity of renal IRI.

Clinical Researches

Effect of two biological immunologic induction therapies on renal transplant recipients

Li Jian, Xu Yahong, Guo Yu, Ma Xiaoping, Lu Yi, Li Yangbo, Jia Zhigang, Zhao Qihua, Luo Shunwen, Chen Ping

2014, 5(5): 299-303. doi: 10.3969/j.issn.1674-7445.2014.05.008

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Abstract:
Objective To evaluate efficacy and safety of two different biologicals in immunologic induction therapy on renal transplant recipients. Methods Clinical data of 78 renal transplant recipients who received biological immunologic induction therapy in Department of Urology of the 452^nd Hospital of Chinese People's Liberation Army from June 2008 to April 2013,were retrospectively analyzed. According to different biological immunologic induction therapy, the recipients were divided into two groups, monoclonal antibody group(group A, n=35, received treatment of basiliximab) and polyclonal antibody group [group B, n=43, received treatment of antithymocyte globulin(ATG)]. And the other recipients who didn't receive immunologic induction therapy in the hospital during the corresponding period were chosen as control group(group C, n=32). The survival rates of recipient and kidney in 3 groups at 12^th weeks after transplantation were analyzed. The levels of serum creatinine(Scr) of recipients in 3 groups were monitored at 7, 14, 30, 60 d after transplantation. The occurrence rates of complications including acute rejection, delayed graft function and infection were compared among 3 groups. Results At 12^th weeks after transplantation, the survival rates of recipient and kidney in 3 groups were 100% and 100% in group A, 97.7% and 97.7% in group B, 100% and 96.9% in group C. There was no significant difference among 3 groups(all in P>0.05). Compared with group C, the Scr levels in group A and B decreased significantly at 7, 14 d after transplantation(all in P<0.05). Compared with group C, the incidence rates of acute rejection decreased in group A and B(all in P<0.05). There was no significant difference in the incidence rates of delayed graft function among 3 groups(all in P>0.05). The incidence rate of infection in group B was significantly higher than those in group A and C(all in P<0.05). Conclusion Immunologic induction therapy is safe and effective in the application on renal transplant recipients.

Value of ordinary color Doppler flow imaging in the diagnosis of hepatic artery thrombosis in early stage after pediatric liver transplantation

Xu Hui, Ren Xiuyun, Yue Yang, Wu Fengdong, Li Wei, Chen Xinguo, Shen Zhongyang

2014, 5(5): 304-307. doi: 10.3969/j.issn.1674-7445.2014.05.009

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Abstract:
Objective To study the value of ordinary color Doppler flow imaging (CDFI) in the diagnosis of hepatic artery thrombosis(HAT) in early stage after pediatric liver transplantation(PLT). Methods Clinical data of 55 children undergoing PLT in the General Hospital of Chinese People's Armed Police Forces from April 2005 to May 2014 were analyzed retrospectively. Conventional hemodynamic monitoring was performed in all cases in the early stage after operation by CDFI. No intrahepatic arterial blood flow signal was observed by CDFI multi-angle examination and HAT was suspected. Computed tomography angiography (CTA) and surgical exploration were performed in the suspected HAT patients. The sensitivity and specificity of ordinary CDFI in diagnosing HAT were calculated by Chiss statistical software. Results Among the 55 PLT recipients, 3 cases were suspected as HAT by CDFI, including 2 cases of HAT in the main hepatic artery, 1 cases of HAT in the right hepatic artery. All cases were confirmed by CTA or surgical exploration. The incidence of HAT was 5% (3/55). No false positive HAT was observed. The sensitivity and the specificity of CDFI in diagnosing HAT after PLT were both 1.0, and false positive rate was 0. Conclusion Ordinary CDFI is the preferred and main method for hemodynamic examination after PLT. The experienced sonographer for transplantation can improve the accuracy in diagnosing HAT.

Research on the oral nucleoside analogues monotherapy in the prevention of hepatitis B recurrence of patients after liver transplantation

Chen Huanwei, Deng Feiwen, Zhen Zuojun

2014, 5(5): 308-313. doi: 10.3969/j.issn.1674-7445.2014.05.010

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Abstract:
Objective To explore the efficacy and safety of the oral nucleoside analogues monotherapy in the prevention of hepatitis B recurrence of patients after liver transplantation (LT). Methods Clinical data of 32 patients with hepatitis B-related disease undergoing LT in the First People's Hospital of Foshan in Guangdong Province from October 1999 to April 2014 were retrospectively analyzed. The patients were divided into two phase groups according to the source of donors. Phase 1 was from October 1999 to September 2007 when 6 patients receiving LT of non-heart-beating donors. The serum hepatitis B virus(HBV) markers of 6 donor livers were all negative. The serum hepatitis B surface antigen (HBsAg) and hepatitis B core antibody (anti-HBc) of the recipients before operation were all positive, including 2 cases combined with Hepatitis B e antigen (HBeAg) positive, 1 case combined with hepatitis B virus e antibody (anti-HBe) positive. The serum HBV deoxyribonucleic acid (DNA) of 5 recipients before LT were over 1 000 copies/ml and 1 case were below 1 000 copies/ml. All the patients in phase 1 group were given lamivudine (100 mg/d) monotherapy orally for the prevention of hepatitis B recurrence after LT. Phase 2 was from November 2011 to April 2014 when 26 patients receiving LT of donation after cardiac death including 1 case of combined liver-kidney transplantation. Six of the donor livers were with serum HBsAg positive and 20 cases negative. Fifteen cases were with hepatitis B surface antibody (anti-HBs) positive and 2 cases with HBeAg positive, 14 cases with anti-HBc positive and 5 with anti-HBe positive. The serum HBV DNA of 11 recipients before LT were over 500 copies/ml and 15 cases below 500 copies/ml. Twenty-five cases were given entecarvir 0.5 mg/d and 1 case telbivudine 600 mg/d monotherapy orally for the prevention of hepatitis B recurrence. Results The median follow-up time of the recipients in group phase 1 was 104 months. The serum HBsAg and HBV DNA were both negative in all recipients and no hepatitis B recurrence was observed till now. The median follow-up time of the recipients in group phase 2 was 50 weeks. Twenty cases received donor livers of negative HBsAg, in which 1 case had transient positive HBsAg 39 weeks after LT and became negative later. The patient receiving combined liver-kidney transplantation suffered hepatitis B recurrence 28 weeks after LT but HBV DNA was observed negative. No hepatitis B recurrence was observed in the 15 cases receiving donor livers of positive anti-HBc. Six cases receiving donor livers of positive HBsAg failed to become negative HBsAg after LT. All the follow-up recipients survived. No HBV DNA replication was observed in the recipients after LT. No adverse reaction of related nucleoside analogues was observed. Conclusion It is effective and safe to use nucleoside analogues monotherapy for the prevention of hepatitis B recurrence in patients after LT.

Short Article

2014, 5(5): 314-317,320. doi: 10.3969/j.issn.1674-7445.2014.05.011

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Abstract: