器官移植

Abstract:
Objective To investigate whether FAM234A is a novel non-Gal antigen in xenotransplantation. Methods Cynomolgus monkeys were immunized with primary porcine aorta endothelial cells (PAEC) from α-1, 3-galactosyltransferase gene-knockout (GTKO) minipigs.The serum samples of immunized monkeys were incubated with PAEC.After immunization, the monkey anti-pig cell antibodies were able to recognize and bind with the unknown antigens on the PAEC surface.Flow cytometry was adopted to determine the mean fluorescence intensity (MFI) of monkey anti-pig cell antibodies binding with the PAEC surface to characterize the content of PAEC surface antigen.Lentivirus-mediated short hairpin ribonucleic acid (shRNA) was utilized to knock down the FAM234A gene in PAEC to detect whether the content of the cell-bound monkey anti-pig cell antibody was reduced and determine if this gene was a potential transplant antigen. Results The non-Gal antigen in PAEC could trigger a large quantity of monkey anti-pig cell antibodies in monkeys.After knock down of the FAM234A gene with shRNA in the PAEC of the GTKO miniature pigs, the monkey IgG antibody binding with PAEC was declined. Conclusions FAM234A is a novel non-Gal antigen in xenotransplantation.

Effect of common immunosuppressants on Tfr cells and Breg cells

Yu Tao, Chen Wen, Liu Zhijia, Huang Haiyan, Xiao Li, Shi Bingyi

2018, 9(5): 360-364，404. doi: 10.3969/j.issn.1674-7445.2018.05.007

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Abstract:
Objective To investigate the effect of common immunosuppressants on the follicular regulatory T cells (Tfr cells) and regulatory B cells (Breg cells). Methods Peripheral blood samples were collected from 6 healthy volunteers (5 mL per case).The lymphocytes were isolated by human lymphocyte isolation solution and cultured.The cells were divided into the control group (without supplement of immunosuppressants), ciclosporin group (250 ng/mL), tacrolimus group (10 ng/mL) and sirolimus group (10 ng/mL).The cells were stimulated with corresponding immunosuppressants for 48 h, respectively.The proportion of Breg and Tfr cells in the lymphocytes in each group was detected by flow cytometry.The content of interleukin (IL)-21 in the supernatants of each group was detected by enzyme-linked immunosorbent assay (ELISA). Results Compared with the control group, the proportion of Breg cells in the lymphocytes in the ciclosporin, tacrolimus and sirolimus groups was all significantly increased (all P < 0.05).In the ciclosporin group and tacrolimus group, the proportion of Tfr cells in the lymphocytes was both significantly elevated (both P < 0.05), whereas the proportion of Tfr cells in the lymphocytes in the sirolimus group was significantly decreased (P < 0.05).ELISA demonstrated that compared with the control group, the IL-21 content in the supernatants in the ciclosporin and tacrolimus groups was both significantly decreased (both P < 0.05).Use of sirolimus exerted no significant effect upon the secretion of IL-21 (P > 0.05). Conclusions Ciclosporin and tacrolimus can inhibit the immune response of B cells.

Effect of warm ischemia time of donor liver from donation after cardiac death on transplant liver in rats

Huang Zhaoyu, Wu Ruichao, Ran Jianghua, Liu Junhan, Zhang Li, Shi Junli, Zhou Tao, Zheng Kepu, Li Wang, Zhang Xibing, Li Yuzhi

2018, 9(5): 365-371. doi: 10.3969/j.issn.1674-7445.2018.05.008

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Abstract:
Objective To evaluate the effect of warm ischemia time of donor liver from donation after cardiac death (DCD) on transplant liver in rats. Methods The rat models of orthotopic liver transplantation of DCD donor liver were established.According to the time of cardiac arrest before obtaining the donor livers, 54 recipient rats were evenly divided into three groups:control group (W0 group, no warm ischemia, n=18), 10 min warm ischemia group (W10 group, n=18) and 20 min warm ischemia group (W20 group, n=18).At 1, 3, and 7 d after operation, serum liver function [alanine aminotransferase (ALT) and aspartate aminotransferase (AST)] were quantitatively measured in rats of each group, liver cells and its organelles were observed by electron microscopy and the expression levels of cytochrome C (Cyt C) and apoptosis inducing factor (AIF) proteins in liver tissues were detected by Western blot in rats of each group. Results The serum levels of ALT and AST in each group were significantly increased at 1 d after liver transplantation along with the prolongation of warm ischemia time.Compared with the W0 group, the serum levels of ALT and AST in the W10 and W20 groups were increased significantly (all P < 0.05).At postoperative 3 d, the serum levels of ALT and AST in each group were slightly decreased.Compared with the W0 group, the serum levels of ALT and AST in the W10 and W20 groups were significantly higher (all P < 0.05).At 7 d after operation, the serum levels of ALT and AST in each group were elevated again.Compared with the W0 group, the serum levels of ALT and AST in the W10 and W20 groups were remarkably increased (all P < 0.05).Electron microscope showed that along with the prolongation of warm ischemia time, the degree of hepatocyte edema was gradually aggravated, the endoplasmic reticular expansion was steadily increased and the degree of mitochondrial damage was aggravated, especially in the W20 group.At postoperative 1 d, the expression level of AIF and Cyt C proteins in the liver tissues did not significantly differ among three groups (all P > 0.05).At 3 and 7 d after operation, compared with the W0 group, the expression levels of AIF and Cyt C proteins were significantly up-regulated in the W10 and W20 groups (all P < 0.05), and the expression levels of AIF and Cyt C proteins in the W20 group were significantly higher than those in the W10 group (both P < 0.05). Conclusions The warm ischemia of the DCD donor liver mainly leads to liver cell injury.Along with the extension of warm ischemia time, significant changes occur in the morphology of mitochondria and other organelles and the expression level of mitochondrial apoptosis-related proteins in transplanted liver cells.

Experimental study of protective effect of hypothermic reperfusion on renal ischemia-reperfusion injury in rabbits

Ding Handong, Liao Guiyi, Zhong Jinbiao, Zhao Fei

2018, 9(5): 372-378. doi: 10.3969/j.issn.1674-7445.2018.05.009

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Abstract:
Objective To explore a novel method for establishing rabbit models with renal ischemia-reperfusion injury (IRI) under hypothermic, normothermic and hyperthermic environments and evaluate the effect of the reperfusion under hypothermic environment on renal IRI in rabbits. Methods Sixty healthy New Zealand rabbits were randomly and evenly divided into five groups:control group (A group, n=12), sham operation group (B group, n=12), hypothermic reperfusion group (C group, n=12), normothermic reperfusion group (D group, n=12) and hyperthermic reperfusion group (E group, n=12).The serum creatinine (Scr) and blood urea nitrogen (BUN) levels were measured daily within postoperative 7 d.The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in the renal tissues were measured at postoperative 1 d.Hematoxylin-eosin (HE) staining was performed to observe the histopathological changes at postoperative 1 d.Terminal deoxynucleotidyl transferase (TdT)-dUTP nick-end labeling (TUNEL staining) was adopted to detect the cellular apoptosis at postoperative 1 d. Results At 1 d after operation, the Scr and BUN levels in C, D and E groups were significantly higher than those in A and B groups (all P < 0.01).Compared with C group, the Scr and BUN levels were more notably increased in D and E groups (both P < 0.05).Within 7 d after surgery, the Scr and BUN levels tended to decline in C, D and E groups.Compared with D and E groups, the Scr and BUN levels were significantly lower in C group (both P < 0.05).Compared with A and B groups, the MDA contents were considerably increased, whereas the SOD activities were significantly decreased in C, D and E groups (all P < 0.01).Compared with C group, the MDA content was more significantly enhanced, whereas the SOD activity was remarkably decreased in D and E groups (both P < 0.01).At postoperative 1 d, the pathological examination of the renal tissues demonstrated that the renal morphology was normal in A and B groups, whereas significant renal damage was observed in D and E groups.Compared with D and E groups, the severity of renal injury was less in C group.TUNEL staining revealed that the quantity of positive renal tubular epithelial cells was significantly increased in D and E groups, and positive cells were also seen in the lumen.The quantity of positive cells in C group was significantly lower than those in D and E groups. Conclusions The reperfusion rabbit models under different temperature environments can be established by covering the kidney by ice soil and persistent administration of 37 ℃ and 40 ℃ saline.Hypothermic reperfusion exerts a protective effect on renal IRI.

Original Articles·Clinical Researches

CRRT combined with immunoabsorption and plasma exchange in treatment of AMR complicated with multiple organ failure after renal transplantation

Sun Kai, Liu Yongbao

2018, 9(5): 379-384. doi: 10.3969/j.issn.1674-7445.2018.05.010

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Abstract:
Objective To investigate the clinical efficacy of continuous renal replacement therapy (CRRT) combined with immunoabsorption and plasma exchange in the treatment of antibody-mediated rejection (AMR) complicated with multiple organ failure after renal transplantation. Methods Twenty-eight patients diagnosed with AMR complicated with multiple organ failure after renal transplantation were randomly divided into the control group (n=12) and treatment group (n=16).All patients were given with methylprednisolone 0.8 g/d shock therapy for 5 d, oral intake of ciclosporin 1.5 mg/(kg·d), CRRT, cardiotonics, diuretics and other symptomatic treatments.In the treatment group, immunoabsorption and plasma exchange were delivered on the 7^th and 10^th d after treatment.The overall conditions were statistically compared between two groups.The changes of renal function [blood urea nitrogen (BUN), serum creatinine (Scr)] and blood electrolyte before and after treatment were compared between two groups.The results of histopathological examination and immunohistochemical staining of the renal tissues before and after treatment were analyzed in two groups. Results Among 28 patients, 24 cases were clinically cured, including 14 in the treatment group and 10 in the control group.The symptoms of 3 patients were deteriorated, including 2 in the treatment group and 1 in the control group.One patient in the control group died of cerebral hemorrhage.After 7 d and 12 d treatment, the levels of BUN and Scr significantly differed between the treatment group and control group (P < 0.05-0.01).The levels of blood sodium and potassium significantly differed between two groups before and after treatment (P < 0.05-0.01).Pathological examination of renal tissue demonstrated that after 12 d treatment, the number of lymphocytes infiltrated in the transplanted kidney tissues was reduced and the degree of vasculitis was alleviated in two groups.Compared with the control group, the pathological scores of glomerulus, tubulointerstitial and blood vessels were significantly lower in the treatment group (all P < 0.01).Immunohistochemical staining revealed that after 12 d treatment, the number of ED1-positive monocytes in the transplanted kidney tissues was declined in both groups, especially in the treatment group.Following 12 d treatment, the ratio of CD4⁺/CD8⁺ cells in renal tubulointerstitial in the treatment group was significantly lower than that in the control group (P < 0.01). Conclusions CRRT combined with immunoabsorption and plasma exchange is an effective method to treat AMR complicated with multiple organ failure following renal transplantation.

Clinical analysis of the influence of gender on blood concentration of mycophenolic acid in recipients undergoing renal transplantation

Zeng Weisheng, Li Zhen, Song Mi, Zhang Changsheng, Zhu Min, Zeng Yunjun

2018, 9(5): 385-389. doi: 10.3969/j.issn.1674-7445.2018.05.011

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Abstract:
Objective To investigate the influence of gender on the blood concentration of mycophenolic acid (MPA) in the recipients after renal transplantation. Methods A total of 115 recipients receiving living donor renal transplantation were recruited and divided into the S1 (61 male cases) and S2 groups (54 female cases).After the operation, triple immunosuppressive regimen of tacrolimus (2 mg for each time, twice daily) + mycophenolate mofetil (MMF, 0.75 g for each time, twice daily) + prednisone was administered for at least one week.The trough blood concentration of MPA was statistically compared between two groups at 1-, 2-, 3-week, 1-, 2- and 3-month after drug administration, and the incidence of adverse reactions were analyzed between two groups. Results At 1-, 2-, 3-week, 1-, 2- and 3-month after drug administration, the trough blood concentration of MPA in the S1 group was significantly lower than that in the S2 group (all P < 0.05).At different time points after drug administration, the ratio of the mean trough blood concentration of MPA between the S1 and S2 groups was relatively stable and fluctuated between 0.71 and 0.84.The percentage of recipients with trough blood concentration of MPA < 1.0 μg/mL in the S1 group was high, especially at 1 week (44%) or 2 week (20%) after drug administration.In the S2 group, the proportion of recipients with trough blood concentration of MPA > 3.5 μg/mL was high with the fluctuations ranging from 30% to 78%.In the S1 group, 4 cases suffered from acute rejection, in whom 3 patients had graft failure due to acute rejection.In the S2 group, 7 patients presented with gastrointestinal disorder and 3 had pulmonary infection. Conclusions Gender exerts certain influence on the trough blood concentration of MPA in the recipients after renal transplantation.The trough blood concentration of MPA in the female recipients is significantly higher than that in the male recipients.

Transverse rectus abdominis myocutaneous flap transplantation with lymphatic tissues for breast reconstruction in treatment of upper limb lymphedema:report of one case with literature review

Chen Ru, Luan Jie, Mu Dali, Liu Chunjun, Xin Minqiang, Mu Lan

2018, 9(5): 390-394. doi: 10.3969/j.issn.1674-7445.2018.05.012

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Abstract:
Objective To summarize the clinical experience of breast reconstruction using transverse rectus abdominis myocutaneous flap transplantation with lymphatic tissues in the treatment of the upper limb lymphedema. Methods One patient presenting with lymphedema of the right upper limb at 7 years after radical mastectomy for breast cancer was admitted to the Plastic Surgery Hospital of the Chinese Academy of Medical Sciences.She was diagnosed with axillary lymphatic flow disorders by preoperative lymphography.The transverse rectus abdominis myocutaneous flap with lymphatic tissue was used for chest wall repair, breast reconstruction and lymphatic flow reconstruction, and thoroughly relieved the severe axillary contracture scars.The circumferences of bilateral upper limbs of the patient were monitored postoperatively for long term. Results After the operation, severe edema occurred in the transplant skin flap.The postoperative drainage volume reached 755 mL/d for 5 d, accompanied by multiple postoperative complications, such as atrial fibrillation, hypoproteinemia, hypokalemia and delayed incisional healing.After timely drainage and standardized comprehensive treatment, excellent postoperative outcomes were obtained.The circumference of the affected upper limb was gradually decreased.The patient was stabilized at postoperative 3 weeks.Excellent surgical efficacy was achieved during 5-year follow-up. Conclusions Transverse rectus abdominis myocutaneous flap transplantation with lymphatic tissues for breast reconstruction is an efficacious treatment of the upper limb lymphedema after radical mastectomy for breast camcer.