Mitochondrial transplantation as a promising therapy for mitochondrial diseases

Mitochondrial diseases are a group of inherited or acquired metabolic disorders caused by mitochondrial dysfunction which may affect almost all the organs in the body and present at any age. However, no satisfactory therapeutic strategies have been available for mitochondrial diseases so far. Mitochondrial transplantation is a burgeoning approach for treatment of mitochondrial diseases by recovery of dysfunctional mitochondria in defective cells using isolated functional mitochondria. Many models of mitochondrial transplantation in cells, animals, and patients have proved effective via various routes of mitochondrial delivery. This review presents different techniques used in mitochondrial isolation and delivery, mechanisms of mitochondrial internalization and consequences of mitochondrial transplantation, along with challenges for clinical application. Despite some unknowns and challenges, mitochondrial transplantation would provide an innovative approach for mitochondrial medicine.

Clinical efficacy of atezolizumab plus bevacizumab combined with local therapy for the treat-ment of recurrent hepatocellular carcinoma before liver transplantation / 中华消化外科杂志

Downstaging treatment by local therapy combined with systemic therapy before liver transplantation for patients with recurrent hepatocellular carcinoma (HCC) can control tumor progression and reduce tumor burden, which resulting in reducing the push-out rate of patients during the waiting period for liver transplantation, providing an oncological observation window, enabling patients of beyond Milan criteria downstaged with better survival benefit. The authors introduce the clinical experience of a case with recurrent HCC of beyond Milan criteria who under-went liver transplantation after receiving atezolizumab plus bevacizumab combined with local therapy. Results show the patient achieving pathological complete remission without postoperative rejection and obtaining a good prognosis with life status improved.

Transplante hepático: variação dos níveis sanguíneos de tacrolimo e desfechos de sobrevida, rejeição e óbito / Liver transplantation: tacrolimus blood levels variation and survival, rejection and death outcomes

ABSTRACT BACKGROUND: Immunosuppressive drugs have important role in transplant of solid grafts, it aim avoid episodes of acute and chronic rejection and improving graft survival and patient survival. In Brazil, in 2016, liver transplantation was the third most frequent, with 1,880 transplants performed, of which 150 in Rio Grande do Sul. Several studies evaluated the association between variability in blood levels of immunosuppressive tacrolimus and late acute cellular graft rejection. OBJECTIVE: To investigate the association of tacrolimus blood levels with clinical outcomes late acute cellular rejection, death, patient survival and graft survival in patients undergoing liver transplantation. METHODS: This is a retrospective longitudinal study including patients submitted to adult liver transplantation by the Liver Transplantation Group in the Santa Casa de Misericórdia Hospital of Porto Alegre, from January 2006 to January 2013, and who used tacrolimus as immunosuppressive therapy. RESULTS: Of the 127 patients included in the study, the majority were male (70.1%), 52-60 years old (33.9%) at the transplant. The most frequent causes of liver transplantation in this series were hepatitis C virus and hepatocellular carcinoma (24.4%) and alcohol (15.7%). Thirteen patients had late acute cellular rejection (10.2%); of these, three had two episodes. Regarding severity classification, seven patients had mild late acute cellular rejection. The mean time of rejection after liver transplantation was 14 months (ranging from 8 to 33 months). Overall survival was 8.98 years. Regarding tacrolimus blood levels, 52 patients with a variation ≥2 standard deviations were identified. Of these patients, eight had rejection; however, the association was not significant (P=0.146). A significant association was found between variation ≥2 standard deviations in tacrolimus blood levels and death (P=0.023) and survival (P=0.019). Regarding 5-year follow-up of graft survival, being two standard deviations above increases by 2.26 times the risk of transplanted graft loss, and for each unit of increase of standard deviation of tacrolimus blood levels there is a two-fold increase in the risk of graft loss in 5 years. CONCLUSION: Increased risk of graft loss associated with increased standard deviations of tacrolimus blood levels may indicate the need for more rigorous and prospective monitoring of tacrolimus blood levels.

RESUMO CONTEXTO: Os imunossupressores desempenham importante papel no transplante de órgãos sólidos, com o objetivo de evitar a rejeição aguda e crônica, aumentando o tempo de sobrevida do órgão e do paciente. No Brasil, em 2016, o transplante de fígado foi o 3° mais frequente, com um número de 1.880 transplantes, sendo 150 realizados no Rio Grande do Sul. OBJETIVO: Investigar a associação da variação dos níveis sanguíneos de tacrolimo com os desfechos clínicos, rejeição celular aguda tardia, óbito, sobrevida de paciente e enxerto em pacientes submetidos ao transplante hepático. MÉTODOS: Trata-se de um estudo longitudinal retrospectivo, no qual foram incluídos os pacientes submetidos ao transplante hepático adulto pelo grupo de transplante hepático na Irmandade Santa Casa de Misericórdia de Porto Alegre, no período de janeiro de 2006 a janeiro de 2013, e que fizeram o uso de tacrolimo como terapia imunossupressora. RESULTADOS: Dos 127 pacientes incluídos no estudo, a maioria era do gênero masculino (70,1%), caucasiana (86,4%), com idade entre 52 e 60 anos (33,9%). As associações de causas mais frequentes para transplante hepático foram vírus da hepatite C, carcinoma hepatocelular (24,4%) e álcool (15,7%). Um total de treze pacientes apresentaram rejeição celular aguda tardia (10,2%); destes, três tiveram dois episódios. O tempo médio de rejeição após o transplante hepático foi de 14 meses, variando de 8 a 33 meses. A sobrevida global foi de 8,98 anos. Em relação aos níveis sanguíneos de tacrolimo, foram identificados 52 pacientes com uma variação maior ou igual a dois desvios-padrão. Destes pacientes, oito tiveram rejeição, contudo, a associação não foi significativa (P=0,146). Foi encontrada uma associação significativa entre a variação maior ou igual a dois desvios-padrão nos níveis sanguíneos de tacrolimo com óbito (P=0,023) e sobrevida (P=0,019). Em relação ao acompanhamento de sobrevida do enxerto em cinco anos, estar dois desvios-padrão acima aumenta em 2,26 vezes o risco de perda do enxerto transplantado, e a cada unidade de aumento de desvio-padrão dos níveis sanguíneos de tacrolimo há um aumento de duas vezes no risco de perda do enxerto transplantado em 5 anos. CONCLUSÃO: O aumento do risco da perda do enxerto associado ao aumento da variação dos níveis sanguíneos de tacrolimo pode indicar a necessidade do acompanhamento mais rigoroso e prospectivo dos níveis sanguíneos de tacrolimo.

Innate immunology research of glutaraldehyde-treated xenogenic blood vessels / 中国免疫学杂志

Objective:To conduct a systematic study of the immunologic response of rats to transplanted glutaraldehyde ( GA)-treated porcine blood vessels in vivo.Methods: The experiment was divided into two groups:fresh group and glutaraldehyde-treated group.Twenty cases of fresh and glutaraldehyde-treated porcine pulmonary arteries were subcutaneously embedded in rats.We compared the changes using HE staining and immunohistochemistry.Results:HE staining showed that there were stronger expression on day 12 and day 30 in the fresh group than that in the glutaraldehyde group.There were similar results in morphology in CD68,C3,IgG.The results of integral optical density ( IOD) in immunohistochemistry showed that IOD started rising from day 4 and got the peak on day 12 or day 30 and or fell on day 60.Conclusion: Innate immunity played an important role in the research on xenogenic immunological rejection mechanism.The immunogenicity of glutaraldehyde-treated xenogenic blood vessels is lower than that in fresh blood vessels.However there is still immunogenicity in glutaraldehyde-treated xenogenic blood vessels.We will explore better ways to obviously weaken the rejection.

Research advances in corneal newborn lymphatic vessel and corneal transplantation rejection / 国际眼科杂志(Guoji Yanke Zazhi)

Corneal newborn lymphatic vessels construct the afferent arc of corneal immunological reaction, which play important role in immune response. The corneal transplantation rejection rate rises due to the emergence of new lymphatic vessel which breaks the immunologic mechanism. With the founding of specific marker of lymphatic endothelial cells and research advancing of growth factor of lymphatic vessels, the mechanism, therapy and prevention of corneal immunological rejection reaction of corneal lymphatic vessel have been studied intensively. The graft survival rate has been greatly improved through inhibiting newborn lymphatic vessel.

A novel noninvasive method to detect rejection after heart transplantation

Prompt and accurate detection of rejection prior to pathological changes after organ transplantation is vital for monitoring rejections. Although biopsy remains the current gold standard for rejection diagnosis, it is an invasive method and cannot be repeated daily. Thus, noninvasive monitoring methods are needed. In this study, by introducing an IL-2 neutralizing monoclonal antibody (IL-2 N-mAb) and immunosuppressants into the culture with the presence of specific stimulators and activated lymphocytes, an activated lymphocyte-specific assay (ALSA) system was established to detect the specific activated lymphocytes. This assay demonstrated that the suppression in the ALSA test was closely related to the existence of specific activated lymphocytes. The ALSA test was applied to 47 heart graft recipients and the proliferation of activated lymphocytes from all rejection recipients proven by endomyocardial biopsies was found to be inhibited by spleen cells from the corresponding donors, suggesting that this suppression could reflect the existence of activated lymphocytes against donor antigens, and thus the rejection of a heart graft. The sensitivity of the ALSA test in these 47 heart graft recipients was 100%; however, the specificity was only 37.5%. It was also demonstrated that IL-2 N-mAb was indispensible, and the proper culture time courses and concentrations of stimulators were essential for the ALSA test. This preliminary study with 47 grafts revealed that the ALSA test was a promising noninvasive tool, which could be used in vitro to assist with the diagnosis of rejection post-heart transplantation.

Expression of Cytokines in Acute Heart Transplantation Rejection / 华中科技大学学报（医学）（英德文版）

The expression and changes of local cytokines network were detected in heart transplantation in rats, so as to determine the role of cytokines in the acute rejection of rats of heart transplantation. Allografts were divided into 4 groups (n=12 in each group): group A (control), group B (IL-2 monoclonal antibody-treated), group C (CsA-treated) and group D (IL-2 monoclonal antibody+CsA-treated). Hearts from DA rats were transplanted into a cervical location in Wistar recipients. The local expression of IL-1β, IL-2, CD25, IL-4, IL-5, IL-6, IL-10, TNFα and INFγ was detected at day 1, 3, 5, 7, 9, 11 and 14 by reverse transcription polymerase chain reaction. The results showed that the survival time of allografts was 8.3±1.7, 29.2±7.1 (P＜0.05), 26.4±5.7 (P＜0.05) and 55.0±10.6 (P＜0.01) days respectively in groups A, B, C and D. The expression of IL-1β, IL-4, IL-10and IFNγ was up-regulated, and that of IL-2, CD25, IL-5, IL-6 and TNFα was significantly inhibited in group A; The expression of IL-1β, IL-5, IL-6, IL-10 and IFNγ was up-regulated, and that of IL-2,IL-4 and TNFα was significantly down-regulated in group B; The expression of IL-1β, IL-2, CD25,IL-5, TNFα and IFNγ was up-regulated, and that of IL-4, IL-6 and IL-10 was significantly down-regulated in group C; The expression of IL-14, Il-5, IL-6 and Il-10 was up-regulated, and that of IL-1β, IL-2, CD25, TNFα and IFNγ was significantly down-regulated in group D. In conclusion,cytokines play an important role in the development of acute transplantation rejection. Different cytokines play different roles in different local environments.

Preventing acute rejection after liver transplantation via blockading B7/CD28 costimulatory pathway in rats / 中国现代普通外科进展

Objective:To observe the effect of CTLA-4Ig on preventing acute rejection and the role of costimulatory molecule B7-1 and B7-2 via the model of orthotopic liver transplantation(ROLT)in rats.Methods:Build the model of ROLT(DA-Lewis).All rats were divided into 2 groups randomly.Group A was control group.Group B was injected with CTLA4-Ig into abdominal cavity after 48 hours of operation.Animals were sacrificed on the day 3,5,7,10 after ROLT to detect the changes of serum biochemistry(ALT,TBIL and DBIL).Liver grafts were collected for measuring the expression of B7-1 and B7-2 mRNA by RT-PCR.Results:1)Compared with the control group,the expression of B7-1 and B7-2 mRNA increased significantly(P

Role of Apoptosis in Acute Rejection of Pancreaticoduodenal Transplantation in Rats / 中国普外基础与临床杂志

Objective To investigate the roles of cell apoptosis and the gene expressions of Fas, FasL, bcl-2 and bax in acute rejection of pancreaticoduodenal transplantation and to evaluate the function of duodenum biopsy for early detection of rejection in rats. Methods Wistar and SD rats were divided into two groups: ①Wistar rats that underwent allogenic pancreaticoduodenal transplantation from the organs of SD rats; ②Wistar rats that received homogenic transplantation. The grafts were then harvested on day 3, 5 and 7 after the transplantation, and all graft samples were observed with HE staining and TUNEL was also used to detect apoptotic cells. The expressions of Fas, FasL, bcl-2 and bax were measured by immunochemical method. According to Nakhleh’s score, pathologic features of transplanted pancreas and duodenum were ranged from one to three scores in order. Results The percentage of same or different scores between the pathological scores of pancreas and duodenum in allogenic pancreaticoduodenal transplantation group were 61.1% (11/18) and 38.9% (7/18) respectively, and there were 6 specimens of pancreatic tissue got higher scores with only one higher score for duodenum. There were significant differences of histopathologic rejection scores and apoptotic indices between the two groups, respectively (P

Effects of p38 Mitogen-Activated Protein Kinase on Apoptosis of Small Intestinal Epithelial Cells after Transplantation in Rats / 中国普外基础与临床杂志

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

The Expression Patterns of CD44 and CD45RB on Peripheral Blood T Lymphocytes in the Rejection of Allogeneic Skin Transplantation in Mice / 대한이식학회지

PURPOSE: Until now, the rejection was diagnosed through a biopsy, but this method of diagnosis reflected the advanced tissue damage of the transplanted organ and contained the innate problem of being invasive. Activation of T lymphocytes, which occurs before the overt tissue damage has a pivotal role in rejection. In relation, our research attempted to evaluate the viability of analyzing the surface antigens of the peripheral blood activated T lymphocytes in mice after skin transplantation as a noninvasive and early diagnostic tool for diagnosis of rejection. METHODS: After the mouse's skin was transplanted, the expression patterns of activated T lymphocyte markers, CD44 and CD45RB were analyzed along with T lymphocyte markers, CD3, CD4, and CD8 using flow cytometry. The skins from the tails of allogeneic Balb/c (H2(d)) mice and syngeneic C57BL/6J mice were transplanted to C57BL/6J (H2(b)) mice as test and control groups, respectively. Peripheral blood, which was sampled from the tail every other day from day 3 to day 15 was stained with anti-CD44 (or CD45RB), anti-CD4 (or CD8) and anti-CD3 monoclonal antibodies simultaneously, and analyzed by 3-color FACS. Repeated ANOVA test and Mann-Whitey test were used to analyze the differences between the expression patterns of peripheral blood T lymphocyte surface antigen in the control and test groups (SPSS 8.0). RESULTS: Rejection occurred only in the test group from day 8 to day 13 (median: day 10). Although the proportions of CD3(+)lymphocytes (CD3(+)%), CD4(+)lymphocytes (CD4(+) %), and CD8 lymphocytes (CD8(+)%) showed no difference between the control and test groups, the total number of peripheral blood lymphocytes and the number of CD3(+)lymphocytes (CD3(+)) and CD8(+)lymphocytes (CD8(+)) decreased more sharply in the control group after day 7. The proportion and the number of CD44 CD3(+)lymphocytes, CD44 CD4(+)lymphocytes, and CD44(+) CD4(+) CD3(+)lymphocytes began to increase after day 7, to peak on day 11, and then to decrease, showing a significant difference from those of the control group. The proportion and number of CD44(+) CD3(+)lymphocytes, in particular, showed the most significant difference among these significant markers. The proportion and number of CD44(+) CD8(+) lymphocytes and CD44(+) CD8(+) CD3(+)lymphocytes showed similar trends to those of CD44(+) CD3(+) or CD44(+) CD4(+), but the differences between the subset proportions in control and test groups were statistically insignificant. No significant difference was observed in any subsets of the CD45RB antigen. CONCLUSION: CD44(+)CD3(+) lymphocytes representing activated T lymphocytes increased significantly compared to the control group during the rejection period of skin transplantation. The analysis of the expression patterns of surface antigen CD44 on peripheral blood T lymphocytes using flow cytometry is sensitive, safe, easily repeatable, and controllable, and, therefore, can be considered a promising tool for the diagnosis of rejection. However, the clear change in CD44 occurred between day 9 and day 13, when rejection was observed grossly. Therefore, it is regarded more useful as a screening test or follow-up indicator rather than as an early diagnostic tool.

Continuous observation to rejection following small bowel transplantation in inbred rats / 中国现代普通外科进展

Objective: To explore the values for rejection research of the models through observing rejection rulers following small bowel transplantation(SBT)in Inbred strain rats.Mothods: Heterotopic SBT models were established with Inbred strain F344/RT11 and BN/RT1n by microsurgical technique according to Monchik method,including Isotransplantation(F344→F344,n=8)and allotransplantation(BN→F344, n=8).Results: 1.The mean living time was over 30 days in IT x group ,which was 12 days in AT x group.2.The general conditions have no significant difference on various time points of postoperative day(POD)in IT x group,while pathological features of grafts were concided with diagnosis of nonspecific slight inflammation on POD3 and were the same as the normal grafts on POD 0?5?7 and 9.3.Significant difference of IL-2R? chain and ?-INF mRNA transcription was earlier than pathological feature appearance of rejection.4.The general conditions and pathological features of grafts were concided with diagnosis of mild,moderate and severe actute rejection in AT x group on POD 5?7and 9 respectively.Conclusion:Inbred strain BN to F344 combination is fitter for research on rejection mechanism than F344 to Wistar combination and Outbred strain Wistar to SD combination.

Expression of ICAM-1 and VCAM-1 in human chronic renal allograft rejection / 中华器官移植杂志

To study the mechanism of human chronic renal allograft rejection, kidney tissues were taken from 16 patients with chronic renal allograft rejection and from 5 healthy subjects, and underwent the frazed section staining for ICAM-1 and VCAM-1 to anti-ICAM-1 and anti-VCAM-1 respectively by using immunohistochemistry(ABC).The results showed that there were differ-ent distribution of ICAM-1 and VCAM-1 expression in nomal kidney and renal allograft during chronic rejection.It was suggested that ICAM-1 and VCAM-1 might play an important role in the pathogenesis of human chronic renal allograft rejection.