1.In vitro study of immunocompatibility of humanized genetically modified pig erythrocytes with human serum
Leijia CHEN ; Mengyi CUI ; Xiangyu SONG ; Kai WANG ; Zhibo JIA ; Liupu YANG ; Yanghui DONG ; Haochen ZUO ; Jiaxiang DU ; Dengke PAN ; Wenjing XU ; Hongbo REN ; Yaqun ZHAO ; Jiang PENG
Organ Transplantation 2024;15(3):415-421
Objective To investigate the differences and the immunocompatibility of wild-type (WT), four-gene modified (TKO/hCD55) and six-gene modified (TKO/hCD55/hCD46/hTBM) pig erythrocytes with human serum. Methods The blood samples were collected from 20 volunteers with different blood groups. WT, TKO/hCD55, TKO/hCD55/hCD46/hTBM pig erythrocytes, ABO-compatible (ABO-C) and ABO-incompatible (ABO-I) human erythrocytes were exposed to human serum of different blood groups, respectively. The blood agglutination and antigen-antibody binding levels (IgG, IgM) and complement-dependent cytotoxicity were detected. The immunocompatibility of two types of genetically modified pig erythrocytes with human serum was evaluated. Results No significant blood agglutination was observed in the ABO-C group. The blood agglutination levels in the WT and ABO-I groups were higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (all P<0.001). The level of erythrocyte lysis in the WT group was higher than those in the ABO-C, TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups. The level of erythrocyte lysis in the ABO-I group was higher than those in the TKO/hCD55 and TKO/hCD55/hCD46/hTBM groups (both P<0.01). The pig erythrocyte binding level with IgM and IgG in the TKO/hCD55 group was lower than those in the WT and ABO-I groups. The pig erythrocyte binding level with IgG and IgM in the TKO/hCD55/hCD46/hTBM group was lower than that in the WT group and pig erythrocyte binding level with IgG was lower than that in the ABO-I group (all P<0.05). Conclusions The immunocompatibility of genetically modified pig erythrocytes is better than that of wild-type pigs and close to that of ABO-C pigs. Humanized pig erythrocytes may be considered as a blood source when blood sources are extremely scarce.
2.Research report of living donor kidney harvesting in Bama miniature pigs with six gene modified
Yong XU ; Xiangyu SONG ; Heng’en WANG ; Shujun YANG ; Zhibo JIA ; Hao WEI ; Shengfeng CHEN ; Mengyi CUI ; Yanling REN ; Jiang PENG ; Shengkun SUN
Organ Transplantation 2024;15(2):229-235
Objective To summarize the experience and practical value of living donor kidney harvesting in Bama miniature pigs with six gene modified. Methods The left kidney of Bama miniature pigs with six gene modified was obtained by living donor kidney harvesting technique. First, the ureter was occluded, and then the inferior vena cava and abdominal aorta were freed. During the harvesting process, the ureter, renal vein and renal artery were exposed and freed in sequence. The vascular forceps were used at the abdominal aorta and inferior vena cava, and the renal artery and vein were immediately perfused with 4℃ renal preservation solution, and stored in ice normal saline for subsequent transplantation. Simultaneously, the donor abdominal aorta and inferior vena cava gap were sutured. The operation time, blood loss, warm and cold ischemia time, postoperative complications and the survival of donors and recipients were recorded. Results The left kidney of the genetically modified pig was successfully harvested. Intraoperative bleeding was 5 mL, warm ischemia time was 45 s, and cold ischemia time was 2.5 h. Neither donor nor recipient pig received blood transfusion, and urinary function of the kidney transplanted into the recipient was recovered. The donor survived for more than 8 months after the left kidney was resected. Conclusions Living donor kidney harvesting is safe and reliable in genetically modified pigs. Branch blood vessels could be processed during kidney harvesting, which shortens the process of kidney repair and the time of cold ischemia. Living donor kidney harvesting contributes to subsequent survival of donors and other scientific researches.
3.Predictive value of cardiopulmonary exercise test in the postoperative complications in patients with esophageal cancer: A retrospective cohort study
Mengyi REN ; Jin LI ; Ming ZHANG ; Miao ZHANG ; Min GAO ; Wei CHEN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(08):1148-1155
Objective To explore the predictive value of cardiopulmonary exercise test (CPET) combined with clinical indexes in the postoperative complications. Methods The clinical data and CPET data (including lung function) of patients undergoing radical esophagectomy in Xuzhou Central Hospital from January 2018 to March 2022 were collected. Univariate analysis and multivariate logistic regression analysis were used to analyze the meaningful evaluation index for the occurrence of postoperative complications. Results A total of 77 patients with esophageal cancer were included, including 59 (76.6%) males and 18 (23.4%) females aged 47-80 years. There were 42 (54.5%) patients in the non-complication group and 35 (45.5%) patients in the complication group. Univariate analysis results showed that the occurrence of postoperative complications was significantly correlated with age, body mass index (BMI), smoking index, tumor stage, the length of postoperative hospital stay, peak work rate (WRpeak), peak kilogram oxygen uptake (VO2peak/kg), the ventilatory equivalent for carbon dioxide slope (VE/VCO2 slope), forced expiratory volume in the first second/forced vital capacity (FEV1/FVC) and maximum expiratory flow rate (MMEF) (P<0.05). The results of multivariate logistic regression analysis showed that BMI [OR=1.35, 95%CI (1.03, 1.77), P=0.031], peakVO2/kg [OR=0.64, 95%CI (0.45, 0.93), P=0.018], oxygen uptake-anaerobic threshold (ATVO2) [OR=0.66, 95%CI (0.44, 0.98), P=0.044] and VE/VCO2 slope [OR=1.49, 95%CI (1.10, 2.02), P=0.011] were the related indexes of complications after radical resection of esophageal cancer. The sensitivity of BMI, VO2peak/kg, ATVO2/kg and VE/VCO2 slope in predicting postoperative complications was 82.10%, and the specificity was 87.44%, 95%CI (0.744, 0.955). Conclusion BMI, VO2peak/kg, ATVO2/kg and VE/VCO2 slope can be used as predictors for postoperative complications of esophageal cancer.
4.Vascularization of vascular endothelial growth factor and collagen I modified beta-tricalcium phosphate porous scaffolds
Mengyi LU ; Yi REN ; Wanqing HU ; Yuan GUI ; Longcheng ZHANG
Chinese Journal of Tissue Engineering Research 2014;(12):1839-1845
BACKGROUND:The auditory ossicle chain reconstruction is stil an important method to treat conductive deafness. Although a great variety of materials have been applied, the blood supply of otosteon after the implantation is ignored. Moreover, there is no real bone formed.
OBJECTIVE:To observe the angiogenesis of vascular endothelial growth factor and col agen I modifiedβ-tricalcium phosphate porous scaffold which is implanted into the otocyst of guinea pig.
METHODS:Total y 60 guinea pigs were randomly divided into experimental group (vascular endothelial growth factor and col agen I modifiedβ-tricalcium phosphate porous scaffold), col agen I control group (col agen I modifiedβ-tricalcium phosphate porous scaffold) and blank control group (β-tricalcium phosphate porous scaffold). The guinea pigs were executed under anesthesia at weeks 1, 2, 3, 4 respectively. The surface of scaffolds was observed by scanning electron microscopy. The angiogenesis of scaffolds were observed by hematoxylin-eosin staining and CD34 immunohistochemistry staining, and then the microvascular density was counted. The osteogenesis of the scaffolds was observed by toluidine blue staining.
RESULTS AND CONCLUSION:Endothelial cel proliferation and lumen formation could be observed after 1 week in the experimental group, and the angiogenesis reach the peak after 3 weeks with traffic branches formedbetween micropores. In the other two groups, the lumen formed at 2 weeks but no traffic branches were visible. The sprouting of new blood vessels in the pores were observed more in the experimental group than the other two groups (P<0.05). The adherence and proliferation of cel s could be examined in the surface and pores of the scaffold by scanning electron microscope. After 4 weeks, the osteogenesis could be observed by toluidine blue staining, especial y in the experimental group. These findings suggest that the vascular endothelial growth factor and col agen I modifiedβ-tricalcium phosphate porous scaffold can realize an effective vascularization in the environment of guinea pigs’ middle ear. What’s more, the scaffold also can promote bone formation.

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