RESUMO
Pigs are considered as ideal donors for xenotransplantation because they have many physiological and anatomical characteristics similar to human beings. However, antibody-mediated immunity, which includes both natural and induced antibody responses, is a major challenge for the success of pig-to-primate xenotransplantation. Various genetic modification methods help to tailor pigs to be appropriate donors for xenotransplantation. In this study, we applied transcription activator-like effector nuclease (TALEN) to knock out the porcine α-1, 3-galactosyltransferase gene GGTA1, which encodes Gal epitopes that induce hyperacute immune rejection in pig-to-human xenotransplantation. Meanwhile, human leukocyte antigen-G5 gene HLA-G5, which acts as an immunosuppressive factor, was co-transfected with TALEN into porcine fetal fibroblasts. The cell colonies of GGTA1 biallelic knockout with positive transgene for HLA-G5 were chosen as nuclear donors to generate genetic modified piglets through a single round of somatic cell nuclear transfer. As a result, we successfully obtained 20 modified piglets that were positive for GGTA1 knockout (GTKO) and half of them expressed the HLA-G5 protein. Gal epitopes on the cell membrane of GTKO/HLA-G5 piglets were completely absent. Western blotting and immunofluorescence showed that HLA-G5 was expressed in the modified piglets. Functionally, the fibroblasts from the GTKO/HLA-G5 piglets showed enhanced resistance to complement-mediated lysis ability compared with those from GTKO-only or wild-type pigs. These results indicate that the GTKO/HLA-G5 pigs could be a valuable donor model to facilitate laboratory studies and clinics for xenotransplantation.
Assuntos
Animais , Humanos , Animais Geneticamente Modificados , Técnicas de Inativação de Genes , Antígenos HLA , Técnicas de Transferência Nuclear , Suínos , Transplante HeterólogoRESUMO
Objective@#To investigate the risk factors of prolonged postoperative length of stay (LOS) in patients with gastric cancer.@*Methods@#A retrospective study was performed on 2033 patients who underwent curative surgery in Department of Pancreatic-gastric, Cancer Hospital, Chinese Academy of Medical Sciences. Eightieth percentile of postoperative hospital stay for all patients was taken as the cutoff, the patients were divided into the normal group (1 532 patients) and the extended group (501 patients). Logistic regression model was used to determine the risk factors related to prolonged postoperative LOS in gastric cancer.@*Results@#A total of 417 cases with postoperative complications were recruited in extended group. The five highest complications accounting for prolonged LOS were: abdominal infection in 69 cases (13.77%), pleural effusion in 60 cases (11.98%), anastomotic leakage in 43 cases (8.58%), poor wound healing in 34 cases (6.79%), and bleeding in 25 cases (4.99%). Logistic regression analysis showed that age (OR=1.348, P=0.013), surgical procedure (OR=2.712, P<0.001), extent of resection (OR=2.035, P<0.001), degree of incision healing(OR=4.867, P<0.001), and perioperative blood transfusion (OR=1.711, P<0.001) were independent risk factors for prolonged postoperative LOS.@*Conclusions@#The most common complication associated with prolonged postoperative LOS for gastric cancer patients is abdominal infection. Age, surgical procedure, extent of resection, degree of incision healing, and perioperative blood transfusion are the independent risk factors for prolonged postoperative LOS.
RESUMO
Objective To investigate the effect of sevoflurane anesthesia on the expression of hippocampal apolipoprotein E (ApoE) mRNA of rats.Methods Sixty healthy male Sprague-Dawley rats,weighing 300-400 g,aged 15 weeks,were randomly divided into 2 groups (n=30 each) using a random number table:50％ oxygen group (group C) and 3.2％ sevoflurane group (group S).The rats in group C inhaled 50％ oxygen for 2 h,while those in group S inhaled 3.2％ sevoflurane in 50％ oxygen for 2 h.Morris water maze test was carried out before anesthesia and at 24 and 72 h after anesthesia.At 2,24 and 72 h after anesthesia,hippocampal specimens were obtained for determination of the expression of ApoE mRNA by RT-PCR.Results Compared with group C,no significant change was found in the escape latency,time of staying at the original platform quadrant,frequency of crossing the original platform,and swimming distance before and after anesthesia,and the expression of ApoE mRNA was up-regulated after anesthesia in group S.There was no significant difference in the escape latency,time of staying at the original platform quadrant,frequency of crossing the original platform,and swimming distance before and after anesthesia,and expression of ApoE mRNA at each time point after anesthesia between the two groups.Conclusion Cognitive dysfunction induced by sevoflurane has no relationship with the up-regulated expression of ApoE mRNA in the hippocampus of rats.
RESUMO
Objective To evaluate the effects of repeated sevoflurane anesthesia on long-term cognitive function in lactating rats.Methods Twenty-four healthy Spragne-Dawley rats,aged 7 days,weighing 14-17 g,were randomly divided into 3 groups (n =8 each):control group (group C),2.6 ％ sevoflurane group (group S1),and 1.5％ sevoflurane group (group S2).At 7,14 and 21 days after birth,2.6％ and 1.5％ sevoflurane and carrier gas were inhaled for 1 h in groups S1,S2 and C,respectively.Visible plafform trial was carried out on 28 and 29 days after birth,and the swimming speed of the rats was recorded.Place navigation test was performed on 32-36 days after birth,and the escape latency was recorded.Spatial probe test was carried out on 36 days after birth,and the time spent in the platform quadrant,swimming distance and the number of times crossing the platform quadrant was recorded.Results Compared with group C,the escape latency was significantly prolonged in groups S1 and S2 (P ＜ 0.05).The escape latency was significantly longer in group S1 than in group S2 (P ＜0.05).There were no significant differences between the three groups in the swimming speed,time spent in the platform zone,swimming distance and the number of times crossing the platform quadrant (P ＞ 0.05).Conclusion Repeated inhalation of sevoflurane during lactation can impair the long-term declarative memory,which is concentration-related,while it has no effects on the associative learning in rats.