器官移植

2016 Vol. 7, No. 3

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Editorial
Experimental Researches
Clinical Researches
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Editorial

Progress on the ultrasonic diagnosis of biliary complications after liver transplantation

2016, 7(3): 167-170. doi: 10.3969/j.issn.1674-7445.2016.03.001

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Experimental Researches

Effect of B cell-derived TGF-β on regulating antibody-induced immune tolerance after transplantation

Zhao Gaoping, Yang Maozhu, Li Shurong, Wei Lingling, Deng Shaoping

2016, 7(3): 171-176. doi: 10.3969/j.issn.1674-7445.2016.03.002

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Objective To investigate the regulatory function of transforming growth factor(TGF)-β in transplantation immune tolerance induced by anti-CD45RB antibody or in combination with anti-Tim-1 antibody. Methods Mouse models with allogeneic cardiac and pancreatic islet transplantation were established. The animals with allogeneic cardiac transplantation were treated with anti-CD45RB antibody and (or) anti-TGF-β antibody and those with pancreatic islet transplantation were handled with anti-CD45RB antibody in combination with anti-Tim-1 antibody supplemented with/without anti-TGF-β antibody. The long-term survival of transplant graft was observed. B cells from recipients which tolerated by pancreas islet transplantation for long time, were isolated and subsequently infused into the mouse models with islet cell transplantation. The survival of pancreas islet transplant graft was observed after anti-TGF-β antibody therapy. After double-antibody treatment in vitro for 14 d, B cell from recipients were mingled with lipopolysaccharide (LPS) and cultured overnight. The percentage of CD19⁺LAP⁺ cell was quantitatively measured by flow cytometer analysis. Results Six among 9 mice with allogeneic cardiac transplantation survived for a long term after anti-CD45RB antibody therapy. All cardiac grafts developed transplant rejection if use of anti-TGF-β antibody was supplemented. In mouse models with islet cell transplantation, combined use of anti-CD45RB and anti-Tim-1 antibodies induced long-term immune tolerance in 90% of transplant grafts. No transplant graft survived for a long period after anti-TGF-β antibody was delivered. Following B cell infusion, 8 of 9 pancreas islet grafts induced no transplant rejection, whereas yielded transplant rejection immediately after supplementing anti-TGF-β antibody. B cells were stimulated and cultured in vitro. Flow cytometry analysis revealed the proportion of CD19⁺LAP⁺ cells in B cells was significantly elevated (P < 0.0001). Conclusions Immune tolerance induced by anti-CD45RB antibody or in combination with anti-Tim-1 antibody depends on TGF-β, probably regulated by regulatory B cells.

Intraportal injection of human umbilical cord blood-derived mesenchymal stem cells combined with dopamine-2 receptor antagonist accelerates the repairing of bile duct in rabbit models with ischemia-type biliary lesion

Hong Chunhong, Zhao Hui, Chen Yunhao, Zheng Bowen, Gu Shijie, Ren Jie, Yi Shuhong

2016, 7(3): 177-181. doi: 10.3969/j.issn.1674-7445.2016.03.003

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Objective To evaluate the effect of intraportal injection of mesenchymal stem cells (MSC) combined with dopamine-2 receptor antagonist (eticlopride) upon accelerating the repairing of bile duct in rabbit models with ischemia-type biliary lesion (ITBL). Methods Twenty-four New Zealand rabbits were randomly divided into four groups (6 in each group) including group A (sham surgery group), group B (ITBL group), group C (MSC group) and group D (MSC+ eticlopride group). The changes in the liver function parameters were monitored before surgery and 1 d, 4 d, 1-, 2-, 3-and 4-week after operation, respectively. At postoperative 4 weeks, cholangiography and ultrasound angiography were performed and liver tissue was sampled for pathological examination. Results Compared with group A, the levels of postoperative liver function parameters in the groups B, C and D were significantly elevated to varying degree(all P < 0.05). Compared with group B, the liver function parameters in the groups C and D were considerably declined (both P < 0.05), especially the group D. The result of biliary tract ultrasound revealed that the ratio of portal vein diameter/bile duct diameter in groups A to D was (3.2±0.3):1, (1.2±0.2):1, (2.1±0.1):1 and (3.0±0.3):1, respectively. The most severe cholangiectasis was observed in group B, relatively slight cholangiectasis in group C and basically normal outcomes in group D. Ultrasound angiography suggested that the most significant improvement in blood supply of the bile duct walls was noted in group D. Pathological examination revealed that the severity of bile duct lesion was the slightest in group D. Conclusions Intraportal injection of MSC in combination with dopamine-2 receptor antagonist can significantly accelerate the repairing of bile tract lesion in rabbit models with ITBL.

Establishment of rat models with in situ continuous perfusion of kidney

Zhang Su, Wang Cheng, Yang Jun, Luo You, Xiong Hu, Fu Shengjun, Ma Baoliang, Yang Li

2016, 7(3): 182-186. doi: 10.3969/j.issn.1674-7445.2016.03.004

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Objective To explore an approach for establishing rat models with in situ continuous renal perfusion, providing a convenient and reliable animal model for fundamental research of renal transplantation. Methods According to the perfusion access, 30 female rats were randomly divided into three groups. In the right renal artery access group (n=10), wound suture under microscope was not required. In the abdominal aorta access group (n=10), the wound of abdominal aorta puncture was sutured under microscope. In the control group (n=10), the opening of the left renal artery was sutured under microscope, and bilateral kidneys were restored by different methods without affecting systemic circulation. The ovarian vein was utilized as the perfusion output. The success rates of continuous kidney perfusion and 2 h reperfusion were statistically compared among three groups. Results In the right renal artery access group, the success rate of perfusion was 80% and 100% for 2 h reperfusion. In the abdominal aorta access group, the perfusion success rate achieved 100% and 80% for 2 h reperfusion. In the control group, the success rate of perfusion was calculated as 80% and 50% for 2 h reperfusion. Conclusions Three types of rat models with in situ continuous kidney perfusion are successfully established. For microsurgery beginners, the technique of right renal artery access is most simple and reliable, followed by abdominal aorta access approach. For the surgeons with microsurgery skills, static cryopreservation of the right kidney and continuous perfusion preservation of the left kidney are highly recommended.

Research on cardiac injury of brain death in Wuzhishan miniature pig models and its mechanism

Chen Yuchao, Teng Liang, Wang Qingqing, Cao Jinglin, Dou Jian

2016, 7(3): 187-190,209. doi: 10.3969/j.issn.1674-7445.2016.03.005

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Objective To evaluate the effect and mechanism of brain death on cardiac injury in pig models established by gradually increasing intracranial pressure. Methods Sixteen Wuzhishan miniature pigs were randomly divided into the experimental and sham surgery groups. In the experimental group, animal models with brain death were established by increasing intracranial pressure. In the sham surgery group, merely cranial catheterization was performed. The changes in the electrocardiogram(ECG) were monitored before and after surgery. The variations of the serum levels of creatine kinase (CK), creatine kinase isoenzyme (CK-MB) and lactate dehydrogenase (LDH) were quantitatively measured before and at 0, 3, 6, 12 h after surgery Results Along with the duration of brain death, decline of ECG amplitude and elevation in T-wave amplitude were observed. At 0, 3 and 6 h after surgery, the levels of CK in the experimental group were significantly higher than those in the sham surgery group (all P < 0.05). At 6 and 12 h after surgery, the CK-MB levels in the experimental group were considerably higher than those in the sham surgery group (all P < 0.05). At postoperative 0, 3, 6 and 12 h, the levels of CK and CK-MB in the experimental group were enhanced compared with preoperative levels (all P < 0.05). However, the LDH levels at 0, 6 and 12 h after surgery were significantly lower than preoperative levels in the sham surgery group (all P < 0.05) Conclusions Brain death pig models present with specific changes in ECG under the status of brain death, probably related to the myocardial damage.

Establishment of cytometric bead assay combined detection on Cys C and NGAL and its application in CMV-infected patients after renal transplantation

Ma Xihui, Huang Haiyan, Gao Yu, Bi Lili, Kong Xiangrui, Liu Yisong, Xiao Li, Shi Bingyi

2016, 7(3): 191-195. doi: 10.3969/j.issn.1674-7445.2016.03.006

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Objective To establish the method of cytometric bead assay in combined detection of cystatin C (Cys C) and neutrophil gelatinase-associated lipocalin (NGAL) and to preliminarily investigate the changes in the serum levels of Cys C and NGAL in cytomegalovirus (CMV)-infected patients after renal transplantation. Methods The experimental method was established as following. The effect and stability of microsphere capsulation were evaluated. The detection range of Cys C and NGAL concentration was improved. The concentrations of biotinylated antibody and PE-streptavidin were quantitatively measured. The correlation coefficient of the standard curve was calculated. The serum levels of Cys C and NGAL in the control (n=10, healthy person), negative CMV (n=10, CMV-pp65 negative patients after renal transplantation) and positive CMV groups (n=20, CMV-pp65 positive patients after renal transplantation) were detected using this method. Results The method of cytometric bead assay in combined detection of Cys C and NGAL was established successfully. In the control, negative and positive CMV groups, the serum levels of NGAL were 8 759(1 465) pg/ml, 10 472(856)pg/ml and 12 817(4 533)pg/ml with statistical significance (all P < 0.001). No statistical significance was noted in terms of Cys C concentration among three groups (all P > 0.05). Conclusions Combined application of cytometric bead assay detects the standard curve of Cys C and NGAL with high relevance and reproducibility. Preliminary results demonstrates that NGAL concentration is significantly elevated in CMV-infected patients following renal transplantation, hinting that CMV active infection probably leads to acute renal tubular injury.

Clinical Researches

Comparison of application value of three pulmonary infection scoring systems in pulmonary infection after renal transplantation

Li Zhibin, Zhang Geng, Ma Shuaijun, Liu Kepu, Ruan Dongli, Qin Weijun, Yuan Jianlin

2016, 7(3): 196-200. doi: 10.3969/j.issn.1674-7445.2016.03.007

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Objective To compare the performance of PSI, CURB-65 and SMART-COP systems in evaluating the severity and predicting the prognosis of pulmonary infection after renal transplantation. Methods Clinical data of 71 patients with pulmonary infection after renal transplantation in Xijing Hospital from January 2009 to June 2015 were retrospectively analyzed. All patients were divided into the severe (n=27) and mild infection groups (n=44). According to the risk-stratification criteria of three scoring systems, all patients were assigned into the low-risk group and moderate-high risk group. The fatality rate, utilization rate of respirator, occupancy rate of intensive care unit (ICU) and length of hospital stay were statistically compared among different risk groups of three scoring systems. The differences among the scores of three systems were compared between the severe and mild infection groups. The efficacy of three scoring systems in predicting the fatality rate of pulmonary infection patients was assessed by receiver operating characteristic (ROC) curve. Results By using three scoring systems, the fatality rate, utilization rate of respirator and occupancy rate of ICU in the moderate-high risk groups were significantly higher than those in the low-risk group (all P < 0.05). The length of hospital stay in the moderate-high risk group was significantly longer than that in the low-risk group (P < 0.05). The scores of PSI, CURB-65 and SMART-COP systems in the severe infection group were considerably higher than those in the mild infection group (all P < 0.05). The optimal cut-off scores of PSI, CURB-65 and SMART-COP systems were 75, 1.5 and 3.5, respectively. The sensitivity, specificity and AUC of PSI, CURB-65 and SMART-COP systems in predicting the fatality rate were calculated as 0.929, 0.890, 0.909; 0.857, 0.772, 0.844; 0.929, 0.860, 0.941, respectively. Conclusions PSI, CURB-65 and SMART-COP systems can assess the severity of pulmonary infection and predict the fatality rate after renal transplantation. SMART-COP possesses the highest accuracy in predicting the fatality rate. PSI is most sensitive but difficult to implement. CURB-65 is simple and convenient to apply in clinical practice.

Effect of training of organ donation after citizen's death on cognition state of medical staff

Huang Liting, Yang Shunliang

2016, 7(3): 201-204. doi: 10.3969/j.issn.1674-7445.2016.03.008

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Objective To investigate the cognition state of medical staff about organ donation after citizen's death and evaluate the effect of professional training. Methods Nine rounds of professional training courses were delivered to the medical staff from Organ Procurement Organization (OPO) of Fuzhou General Hospital of Nanjing Military Command form March to December 2014. Using cluster sampling method, a self-designed questionnaire was utilized to evaluate the cognition state regarding organ donation after citizen's death before and 1 week after professional training. In total, the recovery rate of valid questionnaires was 95.2%(514/540), and the effect of training program was assessed. Results In terms of the knowledge related to organ donation after citizen's death, the mean scores before and after training were (15.0±3.1) and (19.0±3.2) respectively, and there was significant difference (P=0.0001). Regarding the occupation attitude towards organ donation after citizen's death, the support rate was 69.3% before training, considerably enhanced up to 87.9% after training program(P=0.01). Conclusions Medical staff possess certain knowledge of organ donation after citizen's death. Professional training can elevate their cognition state, enhance their understanding of relevant policy and knowledge and increase the support rate of organ donation after citizen's death.

Analysis on risk factors of pulmonary infection after renal transplantation from organ donation after citizen's death

Zhang Pengpeng, Ming Yingzi

2016, 7(3): 205-209. doi: 10.3969/j.issn.1674-7445.2016.03.009

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Objective To investigate the risk factors of pulmonary infection after renal transplantation from organ donation after citizen's death. Methods Clinical data of 125 donors and 210 recipients admitted to Organ Transplantation Center, the Third Xiangya Hospital of Central South Hospital from April 2010 to December 2014 were retrospectively analyzed. All recipients were divided into the pulmonary infection (n=37) and non-pulmonary infection groups (n=173) according to the incidence of pulmonary infection after renal transplantation. General data and clinical prognosis of the recipients with pulmonary infection were evaluated. Risk factors of the donors (gender, age, maintenance therapy time, infection history) and those of the recipients [gender, age, smoking history, diabetes mellitus history, preoperative levels of hemoglobin, leukocyte, percentage of neutrophilic leukocyte, albumin and serum creatinine, postoperative presence of acute rejection, use of biological agents and preventive application of ganciclovir and compound sulfamethoxazole (SMZ)] were subject to single factor analysis and multi-variate Logistic regression analysis to identify the independent risk factors of pulmonary infection. Results and conclusions Among 37 patients with pulmonary infection, 8 died due to progression to severe pulmonary infection. The independent risk factors of pulmonary infection following renal transplantation from organ donation after citizen's death include maintenance therapy time and infection history for the donors, smoking history, diabetes mellitus history and preventive use of ganciclovir or SMZ for the recipients.

Patent ductus arteriosus ligation and simultaneous double-lung transplantation in the treatment of patent ductus arteriosus and Eisenmenger syndrome

Liu Lei, Ye Shugao, Zheng Mingfeng, Ji Yong, Chen Jingyu

2016, 7(3): 210-214. doi: 10.3969/j.issn.1674-7445.2016.03.010

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Objective To investigate the feasibility of patent ductus arteriosus(PDA) ligation and simultaneous double-lung transplantation in the treatment of PDA and Eisenmenger syndrome and its perioperative treatment scheme. Methods On June 27, 2014, one patient with PDA complicated with Eisenmenger syndrome underwent ligation of PDA and simultaneous bilateral sequential lung transplantation assisted under extracorporeal membrane oxygenation (ECMO) in Wuxi People's Hospital. The lung sample was excised strictly according to the standard procedures and perfused in modified low potassium dextran (LPD) solution, which was prepared by our hospital. Surgical procedures were performed precisely and properly. Intraoperative cold ischemia time of the left lung was 3.3 h and 6.0 h for the right lung. Immunosuppressive regimen of tacrolimus (FK506), mycophenolate mofetil (MMF) and adrenal cortical hormone was adopted. Results ECMO support was discontinued when the patient had stable circulation and favorable oxygenation at postoperative 36 h. The tracheal intubation was successfully removed at 3 d after surgery. The patient suffered from acute rejection at postoperative 7 d and relieved after treatment. At postoperative 14-22 d, sputum bacterial culture yielded positive outcomes and recovered after sensitive antibiotics therapy. The patient survived at 1 year after surgery. The parameters of blood gas analysis and lung function were significantly improved. Chest CT scan displayed bilateral transplant lung explicitly and the diameter of pulmonary arterial trunk was dramatically shortened. Cardiac ultrasound examination revealed complete closure of ductus arteriosus with no residual shunt. Conclusions PDA ligation combined with simultaneous allogeneic double-lung transplantation is a feasible approach in the treatment of Eisenmenger syndrome with right-to-left shunt. Favorable lung excision and protection, precise surgical operation and appropriate postoperative management determine the surgical success.

Experience of quality evaluation and function maintenance of donor liver from organ donation after citizen's death

Fu Binsheng, Yi Shuhong, Tang Hui, Yi Huimin, Meng Wei, Zhang Tong, Jiang Nan, Li Hua, Yang Yang, Chen Guihua

2016, 7(3): 215-218. doi: 10.3969/j.issn.1674-7445.2016.03.011

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Objective To summarize the experience of quality evaluation and function maintenance of donor liver from organ donation after citizen's death. Methods Clinical data of 93 donors and 87 patients undergoing liver transplantation in the Third Affiliated Hospital of Sun Yat-sen University form July 2012 to June 2015 were retrospectively analyzed. The experience of quality evaluation and function maintenance of donor liver was briefly introduced. The procurement and utilization of donor organs and the recovery of recipients undergoing donor liver transplantation were summarized. Results The mean warm ischemia time of donor organ was 8 min (1-12 min). The organ procurement surgery was successfully performed with an average duration of 32 min (20-52 min). In total, 93 donor livers were obtained, 87 utilized for liver transplantation and 6 discarded (3 cases of severe fatty liver disease, 2 of hepatic fibrosis and 1 of hepatic tearing injury). Two patients underwent combined liver and kidney transplantation. The function of liver graft recovered well. Merely 1 case presented with primary liver graft non-function. Conclusions Early and accurate evaluation and maintenance of donor livers from organ donation after citizen's death can enhance the quality of organ graft and improve the transplantation efficacy.

Comparison of safety and tolerance of CEUS, CECT and CEMRI for detection of focal liver lesions

Liu Jia, Li Yuhong, Yin Tinghui, Zou Minhong, Wu Lili, Zheng Rongqin

2016, 7(3): 219-221, 242. doi: 10.3969/j.issn.1674-7445.2016.03.012

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Objective To compare the safety and tolerance of contrast-enhanced ultrasound (CEUS), contrast-enhanced CT (CECT) and contrast-enhanced MRI (CEMRI) in the detection of focal liver lesions(FLLs). Methods Clinical data of 42 patients undergoing hepatic CEUS and CECT or CEMRI in the Third Affiliated Hospital of Sun Yat-sen University during July 2015 were collected. Among them, 37 received CEUS and CECT, and 36 underwent CEUS and CEMRI simultaneously. The contrast agent-induced adverse reaction was observed within 30 min after imaging detection. The severity of discomforts during examination was assessed by visual analog scales (VAS). Results The incidence of adverse reaction caused by CEUS and CEMRI contrast agents was lower than that by the CECT contrast agent (0 vs. 11%). The VAS score of patients undergoing CEUS was significantly lower than those of their counterparts receiving CECT or CEMRI (both P < 0.05). Conclusions CEUS is a preferential option for the detection of FLLs with higher safety and better tolerance.

Comparison of clinical efficacy between tauroursodeoxycholic acid and ursodesoxycholic acid on liver function recovery after liver transplantation

Wang Yue, Zhu Zhijun, Sun Liying, Wei Lin, Qu Wei, Liu Ying, Zeng Zhigui

2016, 7(3): 222-226. doi: 10.3969/j.issn.1674-7445.2016.03.013

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Objective To compare the clinical efficacy between tauroursodeoxycholic acid and ursodesoxycholic acid on liver function recovery following liver transplantation. Methods Twenty eight recipients undergoing orthotopic liver transplantation in Beijing Friendship Hospital Affiliated to Capital Medical University from 2006 to 2012 were recruited in this investigation. All patients were orally administered with ursodesoxycholic acid for 3 months and subsequently received oral administration of tauroursodeoxycholic acid for 6 months. The changes in the levels of total bile acid (TBA), total bilirubin (TB), direct bilirubin (DB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (GGT) were statistically compared before and 1, 2, 3, 4, 5, 6 and 9 months after enrollment. At 3 and 9 months following enrollment, the components of bile acid were determined in 5 cases to compare the changes between the hydrophobic and hydrophilic bile acid. Results Within 3 months after oral administration of ursodesoxycholic acid, all parameters related to liver function were elevated. Compared with the level at 3 months after enrollment, the level of TBA was significantly decreased at 5 and 9 months after enrollment. The level of ALT declined at 5 and 6 months. The concentration of AST was decreased at 4 and 5 months. The largest decrease in the level of ALP was noted at 9 months. The content of GGT was found to decline at 6 and 9 months (all P < 0.05). After oral administration of tauroursodeoxycholic acid, a higher level of hydrophilic bile acid was detected in the patients' bile acid, dominated by tauroursodeoxycholic acid. Conclusions Tauroursodeoxycholic acid exerts higher clinical efficacy on promoting the secretion of bile and protecting liver function compared with ursodesoxycholic acid after liver transplantation.