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BACKGROUND: Searching for a substitute donor corneal is a hotspot for treating fungal corneal ulcer. OBJECTIVE: To Investigate the clinical effect of bio-engineering cornea and donor cornea on treating fungal corneal ulcer. METHODS: Forty-four cases (44 eyes) of fungal corneal in General Hospital of Northern Theater Command were enrolled, and were randomized into two groups, followed by underwent lamellar keratoplasty using acellular porcine corneal matrix (bio-engineering group, n=22) and human donor cornea (donor group, n=22). The patients were followed up for 12 months. The control rate of infection, visual acuity, graft transparency, epithelizatlon time and complications were observed In both groups. The study was approved by the Ethical Committee of General Hospital of Northern Theater Command, approval No. K(2018)05. RESULTS AND CONCLUSION: (1) The control rate of Infection showed no significant difference between two groups (91%, 91%, P > 0.05). (2) The visual acuity in both groups was improved with time. The visual acuity in the donor group was significantly better than that in the bio-engineering group at 12 months after surgery (P 0.05). (4) The epithelization time showed no significant difference [(6.6±2.0) days, (6.7±1.9) days, P > 0.05]. (5) There was no significant difference In the Incidence of delayed healing of corneal epithelium, rejection reaction of graft, neovascularization, or recurrence between two group (P > 0.05). The rate of graft dissolved In the bio-engineering group was significantly higher than that In the donor group (32%, 8%, P < 0.05). (6) In summary, bio-engineering cornea used for lamellar keratoplasty holds significant efficacy, high safety and good prognosis in the treatment of fungal cornea ulcer, which may as substitute when donor cornea is deficient.
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@#Keratoplasty is routinely used to treat end-stage corneal diseases. However, immune-mediated graft rejection remains the major cause of surgical failure. Organ transplant rejection is often due to the directional migration and homing of inflammatory cells to lymphoid tissues and local inflammatory sites, which is regulated by various adhesion molecules and chemokines. Regulatory T cells play a key role in immune regulation and are essential for maintaining peripheral tolerance. Recent studies have revealed that regulatory T cells play important roles in preventing organ transplant rejection and the development of autoimmune diseases. This review will summarize the recent research on the induction of ocular immune privilege by regulatory T cells, with special focus on how regulatory T cells mediate tolerance in the eye and clinical potential of modulating these mechanisms during corneal transplantation.
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Objective To summarize the practice experience of establishing a stable abdominal heart transplantation model combined with tail vein injection in mice. Methods In the preliminary experiment, 50 pairs of donor and recipient Kunming mice received isotransplantation, 40 pairs of donor and recipient C57BL/6J mice underwent isotransplantation. In the formal experiment, 10 pairs of donor and recipient C57BL/6J mice received isotransplantation, 30 pairs of Balb/c mice as the donor and C57BL/6J mice as the recipient received allotransplantation. The time of each step of the heart transplantation (including harvesting and dressing of the donor heart, vascular anastomosis of the recipient, etc.) was recorded. The duration of transplanted heart beat and the survival time of the recipient was observed daily after operation. The time required for tail vein injection in the transplanted mice was recorded. Pathological examination of the transplanted heart was performed at 30 d after isotransplantation (n=5) and 7 d after allotransplantation (n=5). Results In the formal experiment, the success rate of heart transplantation was 90%. The harvesting and dressing time of donor heart was (13.9±0.6) min. The cold ischemia time of the recipient was (14.2±1.2) min. The vascular anastomosis time was (34.2±3.1) min. The total operation time was (86.6±5.4) min. Postoperatively, the transplanted heart of the mice undergoing isotransplantation survived longer than 100 d. Pathological examination at postoperative 30 d demonstrated only a slight amount of inflammatory cell infiltration. The survival time of the mice receiving allotransplantation was (7.2±0.5) d due to rejection reaction. At postoperative 7 d, pathological examination showed a large quantity of inflammatory cells infiltrating into the myocardium, manifested with acute cellular rejection. The success rate reached 90% after over 200 times of tail vein injection. Conclusions In this study, a stable mouse abdominal heart transplantation model is successfully established. The mouse models in the preliminary experiment can be utilized for tail vein injection.
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Objective To investigate the effect of human CD47 (hCD47) in inducing the immune tolerance of human macrophages to porcine endothelial cells. Methods The porcine iliac endothelial cell (PIEC) transfected with pCDH-hCD47-FLAG plasmid was assigned into the pCDH-hCD47 group, PIEC transfected with pCDH-FLAG empty vector plasmid was assigned into the pCDH group, PIEC transfected with hCD47-dN was assigned into the pCDH-hCD47-dN group and human umbilical vein endothelial cell (HUVEC) was assigned into the positive control group. The cells were co-cultured with human macrophages to detect and analyze the phosphorylation of signal regulatory protein α (SIRPα) and the killing effect of human macrophages on PIEC. Furthermore, porcine arteriae endothelial cell (PAEC) was isolated from GT－/－ and GT－/－/ hCD 47 gene editing pigs to analyze the phosphorylation of SIRPα and the killing effect of human macrophages on PAEC. Results The pCDH group cells could not induce the phosphorylation of SIRPα, whereas the pCDH-hCD47 group cells could activate the phosphorylation of SIRPα after 10 min co-culture with human macrophages, and the degree of phosphorylation of SIRPα was increased with the prolongation of the co-culture time. The pCDH-hCD47-dN group cells failed to activate the phosphorylation of SIRPα. Human macrophages exerted significant effect on killing the pCDH group cells. The pCDH-hCD47 group cells could evidently inhibit the killing effect of human macrophages (P < 0.05), whereas the pCDH-hCD47-dN cells failed to suppress the killing effect of human macrophages. GT－/－-PAEC could not activate the phosphorylation of SIRPα after co-culture with human macrophages. However, GT－/－/hCD47-PAEC significantly activated the phosphorylation of SIRPα after co-culture with human macrophages. Human macrophages exerted significant killing effect on GT－/－-PAEC, and GT－/－/hCD47-PAEC could obviously inhibit the killing effect of human macrophages (P < 0.05). Conclusions The expression of hCD47 in the porcine endothelial cells can inhibit the killing effect of human macrophages on endothelial cells by activating the phosphorylation of SIRPα.
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Objective To investigate the effects of adoptive reinfusion of regulatory T cell (Treg) on the recovery of islet function and graft survival time after islet allograft transplantation. Methods The diabetic model was established using C57BL/6 mice as recipients, and Balb/c mice were chosen as donors for islet allografts transplantation beneath the renal capsule. The recipient mice were divided into 3 groups and 3 mice in each group according to different processing Methods: Treg experiment group (Treg group, 1×106 Treg cells were injected via tail vein at 1 d before operation), positive control group [sirolimus (SRL) group, SRL at a dose of 300 μg/(kg·d) was intragastrically given every day from 1 d before operation] and blank control group (control group, an equivalent volume of normal saline was intragastrically given every day from 1 d before operation). Enzyme-linked immune absorbent assay (ELISA) was used to detect the changes of blood glucose and C-peptide in mice within 14 days after transplantation. In vivo imaging technique was used to dynamically monitor the survival of mice within 14 days after transplantation. Results In each group, the blood glucose levels at postoperative 3 d were significantly decreased compared with those before transplantation (all P < 0.001). At postoperative 1 d, the C-peptide levels showed an explosive rise to varying degree in each group. At postoperative 3 d, the C-peptide levels in each group were significantly higher than that before operation (all P < 0.001). At the end of the observation period at 14 d after operation, the C-peptide levels in the SRL and Treg groups were (427±50) pmol/L and (833±57) pmol/L, relatively higher than that in the control group. But the blood glucose levels were (14.5±0.5) mmol/L and (12.1±0.6) mmol/L, significantly lower than that in the control group (all P < 0.001). Compared with the SRL group, the explosive release amount of C-peptide was significantly lower, the declining trend was more remarkably stable, and the C-peptide level was considerably higher in the Treg group at the end of the observation period (all P < 0.001). At postoperative 14 d, the grafts were almost completely apoptotic in the control group, over 50% of the grafts survived in the SRL group, and over 80% of the grafts survived in the Treg group. Conclusions Adoptive reinfusion of Treg cells can effectively protect islet grafts, prolong the survival time of grafts, and maintain the normal levels of blood glucose and C-peptide in the recipient mice.
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Objective To investigate the effect and mechanism of all-trans retinoic acid (ATRA) on the cyclosporin (CsA)-induced proliferation and apoptosis of glomerular mesangial cells in rat models. Methods The glomerular mesangial cells induced by different doses of CsA were treated with different doses of ATRA. MTT assay was carried out to detect cell proliferation. Hoechst 33258 fluorescent staining was adopted to observe the morphology of the apoptotic cells. Flow cytometry was conducted to detect the cellular apoptosis rate. Immunofluorescent staining was employed to quantitatively measure the expression level of mitochondria-derived pro-apoptotic Smac protein. Results Compared with the control group, administration of CsA at a dose of 0.5 μg/mL and above could suppress cellular proliferation, and use of CsA at a dose of 1.0 μg/mL and above could induce cellular apoptosis. The expression level of Smac protein was significantly up-regulated by CsA administration with a dose and time dependence (all P<0.05).Compared with the CsA group, combined administration of CsA and ATRA exerted a more significant inhibitory effect on cellular proliferation. Supplement of ATRA could significantly inhibit glomerular mesangial cellular apoptosis induced by CsA and down-regulate the expression of Smac protein with a dose dependence (both P<0.05). Conclusions CsA can inhibit cellular proliferation, induce cellular apoptosis and up-regulate the expression of Smac protein of glomerular mesangial cells. ATRA is capable of suppressing glomerular mesangial cellular apoptosis induced by CsA, which is probably mediated by the Smac signaling pathway.
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Objective To compare the clinical efficacy between pediatric ABO-incompatible liver transplantation (ILT) and ABO-compatible liver transplantation (CLT) by Meta-analysis. Methods Relevant studies published until May 2017 were electronically retrieved from PubMed, Embase, Cochrane library, China national knowledge internet (CNKI),Wanfang and VIP databases. According to the inclusion and exclusion criteria, the publications eligible were screened and clinical data were extracted. Meta-analysis was performed using the random or fixed effect model analyses with Review Manager 5.3 statistical software. Results Eleven retrospective cohort studies in English were selected. Meta-analysis demonstrated that the postoperative 1-year survival rate of the recipients in the ILT group was significantly lower than that in the CLT group [odds ratio (OR)=0.64, 95% confidence interval (CI) 0.49-0.83, P=0.0007)]. In the ILT group, the incidence of postoperative rejection reactions was considerably higher compared with that in the CLT group (OR=1.96,95% CI 1.03-3.72, P=0.04). No statistical significance was observed in the postoperative 3- and 10-year survival rate of the recipients, 1-, 3- and 10-year survival rate of the graft and the incidence of postoperative biliary tract complications between two groups (all P>0.05). Conclusions Compared with their CLT counterparts, the 1-year survival rate of the ILT recipients is lower, whereas the incidence of rejection reactions is higher. However, the long-term survival rate of both the recipient and graft and the incidence of biliary tract complications do not significantly differ between CLT and ILT. ILT is a relatively ideal option for emergent patients or those lacking of liver graft with compatible blood group for a long period of time.
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Objective To evaluate the effect of extracorporeal photopheresis upon the expression levels of interleukin (IL)-12p70 and T helper cell (Th) 1/Th2-like cytokines in splenic lymphocytes of mouse models undergoing skin transplantation. Methods The C57BL/6 mice were used as the donors and BALB/c mice as the recipients to establish mouse models with skin allograft. The splenic lymphocytes in the C57BL/6 and BALB/c mice (CSP and BSP) were isolated and treated with 8-methoxypsoralen combined long-wave ultraviolet (PUVA-SP). According to the components of intravenous infusion into the recipients, all experimental animals were randomly divided into the PUVA-BSP, PUVA-CSP, BSP, CSP and phosphate buffer solution (PBS) control groups (n=12 for each group). The mice were injected with PUVA-BSP, PUVA-CSP, BSP, CSP or PBS via the caudal vein at preoperative 7 d, upon the day of surgery and at postoperative 7 d, respectively. The apoptosis of the splenic lymphocytes was observed after PUVA treatment. The expression levels of IL-12p70 and Th1/Th2-like cytokines in the peripheral blood of the recipients were quantitatively measured. Results After the skin transplantation, the expression levels of IL-12p70 in the peripheral blood of mice in the PUVA-BSP and PUVA-CSP groups were significantly down-regulated compared with those in the BSP, CSP and PBS control groups (all P<0.01). In the PUVA-BSP and PUVA-CSP groups, the expression levels of Th1-like cytokine IL-2, interferon (IFN)-γwere dramatically lower than those in the BSP, CSP and PBS control groups (all P<0.01). The expression levels of Th2-like cytokine IL-10 in the PUVA-BSP and PUVA-CSP groups were significantly up-regulated compared with those in the BSP, CSP and PBS control groups (all P<0.01). Conclusions Infusion of PUVA-SP at a sufficient dose can induce the low expression level of IL-12p70 and drive the incidence of Th2 immune deviation in the recipient BALB/c mice.
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Objective To systematic evaluation the therapeutic effects on patients with ABO-incompatibility liver transplantation (ILT),and compare the curative effect with ABO-compatible liver transplantation (CLT). Methods The literatures of comparison in clinical efficacy between ILT and CLT were collected at home and abroad by computer search in PubMed database,Embase database,Cochrane database,Medline database,Web of science database,CNKI,Wanfang database,VIP database,et al,and the quality of literatures were accessed. Meta analysis was carried out by fixed effect model and random effect model with RevMan5.3 software. Results A total of 18 papers were included. The results of Meta analysis showed that there was no significant difference in the survival rates of recipient between ILT group and CLT group at 1 ,3 and 5 years after operation (all P>0.05 ). Compared with CLT group,the survival rates of grafts were significantly decreased in ILT group at 1 ,3 and 5 years after operation,and the difference was statistically significant (all P<0.05 ). The incidences of postoperative biliary complication and acute rejection in ILT group were significantly higher than those in CLT group,the difference was statistically significant (both P<0.05 ). Conclusions Compared with CLT, the curative effect of ILT is weaker but still can be used as a new choice for critical condition of the recipient or waiting for the donor liver for a long time.
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Objective To assess the value of two-dimentional strain/strain rate (2D-S/SR) imaging compared with endomyocardial biopsy (EMB) in detecting acute rejection (AR) reaction after heart transplantation (HT). Methods Twenty-five patients who required HT underwent echocardiography within 12 h after EMB. Ten patients of grade 0 AR were regarded as group A, 8 patients with grades Ⅰa-Ⅰb were considered as group B, 7 patients with grades ≥Ⅱa were considered as group C. Thirty age-matched normal controls were considered as group D. The longitudinal, radial and circumferential stain and strain rate of different ventricular wall in systolic were measured and compared among groups. Results Compared with the group D, longitudinal stain (LS) of basal-septum, circumferential strain rate (CSR) of ant-septum at the level of mitral valve, radial strain (RS) of all segments at the level of mitral valve reduced in groups A, B and C. Conclusion The RS of 2D-S/SR is an effective and sensitive technique for the detection of acute rejection, and longitudinal stain can be used as an early index of AR for grade ≥Ⅱ after HT.
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Objective This study was to detect the expression and level of vascular endothelial growth factor(VEGF)in the acute rejection reaction of orthotopic liver transplantation in rats,which attempted to prove whether VEGF is the key molecule mediating the inter-permeate and inter-enhancement mediated by cells between Angiogenesis and inflammation reaction Methods Expressions of VEGF in plasm in liver and spleen were detected using immunohistochemical staining.The levels of VEGF were measured with enzyme linked immunosorbent assay(ELISA).Results The expression of VEGF in liver,spleen and the level of VEGF in plasma in experiment group were higher than that in the control group(P＜0.05),which were the highest in two days after operation.Conclusion VEGF may play a significant role in the acute rejection reaction of orthotopic liver transplantation in rats.There was a close relation between the expression and level of VEGF in liver,spleen and survival time of graft.VEGF was a kind factor which is expressed in early stage.
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Objective To explore the effects of p38 mitogen-activated protein kinase (MAPK) on apoptosis of small intestinal epithelial cells after transplantation in rats. Methods Small intestinal transplantation was performed in SD and Wistar rats. The recipients were divided into three groups: isograft group (Wistar→Wistar group), allograft group (SD→Wistar group) and allograft+cyclosporine A group (SD→Wistar+CsA group). The grafts were harvested on day 1, 3, 5 and 7 after operation. All graft samples were subjected to histological examination. The apoptosis of graft epithelial cells was detected by TUNEL method. p38 MAPK was measured by Western-blotting method and serum TNF-? was determined by ELISA. Results Mild, moderate and severe rejection reaction occurred in the SD→Wistar group, it was showed that the number of apoptotic cells increased with the severity of the rejection reaction by TUNEL. In SD→Wistar group, the numbers of apoptotic cells were significantly higher than those of the other two groups (P0.05). The expression of p38 MAPK and the level of serum TNF-? were positively correlated with apoptosis in small intestinal rejection after transplantation (r=0.875, P
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Heme oxygenase(HO) is a rate-limiting enzyme that catalyzes heme degradation.It catabolizes heme into 3 products: carbon monoxide(CO),free iron and bilirubin,which play an important role in keeping the steady state of the organism.In recent years,some studies show that the promotion of HO-1 expression can reduce ischemia-reperfusion injury of an organ transplant and regulate the transplantation immunological rejection.It has an important application prospect in liver transplantation.
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A heterologous corneal endothelial transplantation was attempted using human endothelial cells and a Lewis rat penetrating keratoplasty model. Cultured human endothelial cells were seeded to a Lewis rat cornea, which was denuded of its endothelium. When grafted into the syngeneic Lewis rat, the graft remained clear for at least five days, and then became opaque and edematous because of immune rejection reaction. In contrast, corneas denuded of their endothelium became opaque and edematous immediately after transplantation. These results demonstrate that transplanted endothelial cells have enough antigens to induce rejection reaction even though they have the functional capacity to deturge the cornea.
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Animals , Female , Humans , Rats , Endothelium, Corneal/cytology , Graft Rejection/immunology , Major Histocompatibility Complex/immunology , Rats, Inbred Lew , Transplantation, HeterologousABSTRACT
An orthotopic penetrating keratoplasty model was developed in the rat. An oversized (0.5 mm) graft was used and 8 interrupted sutures were applied. These sutures were not removed. Eleven grafts out of 13 were rejected by the 3rd week in the disparate group (Brown Norway rat to Lewis rat transplantation group), which was characterized by edema, opacity, and neovascularization. All grafts remained clear in the syngeneic group (Lewis rat to Lewis rat transplantation group). Immunohistochemical examination was performed. This model seems to be a reliable and reproducible one to evaluate rejection reaction in corneal transplantation.