1.Application of xenotransplantation in clinical practice
Shengkun SUN ; Shujun YANG ; Hao WEI ; Haihong YANG ; Jing LU ; Jiang PENG
Organ Transplantation 2024;15(2):200-206
Organ transplantation is the optimal treatment for end-stage organ failure. Nevertheless, organ shortage is a global problem, which limits further development of organ transplantation. Recent research shows that genetically modified pig may become a realistic alternative source of clinical organ transplantation donor. Xenotransplantation may serve as one of the effective measures to resolve the problem of organ shortage. Since 2021, 2 cases of living xenotransplantation and 6 cases of xenotransplantation in brain death recipients have been performed worldwide, and phase Ⅰ clinical trial of xenotransplantation has been launched, and the results have exceeded expectations. Therefore, in this article, recent clinical trial results of xenotransplantation in living and brain death recipients were retrospectively analyzed, and scientific, technical and ethical issues related to clinical research of xenotransplantation were illustrated, hoping to provide reference for clinical research of xenotransplantation in China and promote the development of xenotransplantation in clinical practice.
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.Significance and successful construction of stable transfection of MH7A cells with lower TRAF2 expression using lentiviral vector
Luying CHEN ; Liping JIANG ; Weikang WANG ; Shujun ZUO ; Jiajie KUAI ; Yang MA ; Chenchen HAN ; Wei WEI
Acta Universitatis Medicinalis Anhui 2024;59(2):193-199
Objective To construct a stable synovial cell line MH7A from rheumatoid arthritis(RA)patients using lentiviral vectors that interfere with the expression of tumor necrosis factor receptor associated factor 2(TRAF2),and to study the role of TNF-α-TRAF2 signaling in MH7A abnormal proliferation.Methods Based on the design principles of human TRAF2 gene sequence and shRNA sequence,three pairs of TRAF2 shRNA interference se-quences were designed and synthesized.The primers were annealed by PCR,and a linear vector was obtained by double enzyme digestion PLKO.1-puro.The linearized vector was connected to the annealed primers through Solu-tion I,and the connected products were introduced into receptive cells.The plates were coated,and positive colo-nies were selected for sequencing.Three different recombinant plasmids of PLKO.1-TRAF2-shRNA lentivirus were constructed,and lentivirus packaging plasmids was used to package logarithmic growth phase HEK 293T cells.Vi-rus solution was collected to infect MH7A cells.At the same time,puromycin was used to screen MH7A stable transgenic strains with low TRAF2 expression.CCK-8 method,Western blot,and qPCR were used to detect the proliferation function of MH7A induced by TNF-α and low expression of TRAF2,as well as downstream signal TRAF2,P65 protein expression and mRNA levels.Results PLKO.1-TRAF2-shRNA(1),PLKO.1-TRAF2-shR-NA(2),and PLKO.1-TRAF2-shRNA(3)lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were successfully constructed.The three TRAF2-shRNA lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were respectively introduced into the lentivirus packaging plasmid of HEK 293T to obtain virus solution.After infecting MH7A cells with the virus solution,they were treated with puromycin(2.00 μ G/mL)screening and obtaining MH7A stable transgenic plants after 2 days.Through qPCR and Western blot results,it was found that the expression of TRAF2 mRNA and protein in PLKO.1-TRAF2-shRNA(1)MH7A stably transfected cells was significantly reduced compared to the negative control group.The results of CCK-8 and Western blot showed that after knocking down TRAF2 in MH7A,the proliferation of MH7A cells with low TRAF2 expression induced by TNF-α and the phosphorylation level of P65 were significantly reduced.Conclusion A sta-ble transgenic strain of PLKO.1-TRAF2-shRNA(1)MH7A cells was successfully constructed to investigate the role of TNF-α-TRAF2 signal activation in mediating abnormal proliferation of RA synovial cells.
4.Construction and validation of a prediction model for public acceptance of kidney xenotransplantation in China
Shujun YANG ; Hao WEI ; Jiang PENG ; Mengyi CUI ; Panfeng SHANG ; Shengkun SUN
Organ Transplantation 2024;15(1):102-111
Objective To explore the public attitude towards kidney xenotransplantation in China by constructing and validating the prediction model based on xenotransplantation questionnaire. Methods A convenient sampling survey was conducted among the public in China with the platform of Wenjuanxing to analyze public acceptance of kidney xenotransplantation and influencing factors. Using random distribution method, all included questionnaires (n=2 280) were divided into the training and validation sets according to a ratio of 7:3. A prediction model was constructed and validated. Results A total of 2 280 questionnaires were included. The public acceptance rate of xenotransplantation was 71.3%. Multivariate analysis showed that gender, marital status, resident area, medical insurance coverage, religious belief, vegetarianism, awareness of kidney xenotransplantation and whether on the waiting list for kidney transplantation were the independent influencing factors for public acceptance of kidney xenotransplantation (all P<0.05). The area under the curve (AUC) of receiver operating characteristic (ROC) of the prediction model in the training set was 0.773, and 0.785 in the validation set. The calibration curves in the training and validation sets indicated that the prediction models yielded good prediction value. Decision curve analysis (DCA) suggested that the prediction efficiency of the model was high. Conclusions In China, public acceptance of kidney xenotransplantation is relatively high, whereas it remains to be significantly enhanced. The prediction model based on questionnaire survey has favorable prediction efficiency, which provides reference for subsequent research.
5.Expression and prognostic factors analysis of CYFRA21-1, CEA, and Ki67 in non-small cell lung cancer
Jianyun PAN ; Yien HUANG ; Shujun HONG ; Shaohan FANG ; Jingwei LIU ; Weiqiang CHEN ; Gaojian PAN ; Jie JIANG ; Guojun GENG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(05):682-688
Objective To explore the value of preoperative detection of soluble fragments of cytokeratin-19 (CYFRA21-1), carcinoembryonic antigen (CEA), and postoperative detection of nuclear proliferation associated antigen Ki67 in prognostic evaluation of non-small cell lung cancer patients. Methods The clinicopathological data and follow-up results of patients with non-small cell lung cancer treated in the Department of Thoracic Surgery of the First Affiliated Hospital of Xiamen University in 2017 were collected. CYFRA21-1>3.39 ng/mL was defined as positive, and CEA>5 ng/mL was defined as positive. The receiver operating characteristic curve (ROC curve) of Ki67 expression level was drawn. The maximum area under the curve (AUC) was the cutoff value of Ki67 expression level, and the Ki67 expression level greater than its cutoff value was defined as positive. Cox regression analysis was used to determine the independent risk factors for poor prognosis in patients with non-small cell lung cancer. Results Finally 248 patients were collected, including 125 males and 123 females, with a median age of 61 years (ranging from 30 to 81 years) at the time of surgery. Univariate analysis showed that positive CYFRA21-1, high expression of Ki67, positive CEA, age≥60 years at operation, distant metastasis, lymph node metastasis, maximum tumor diameter>3 cm, and TNM stage Ⅲ were associated with poor prognosis in patients with non-small cell lung cancer. When combined detection of preoperative tumor markers and postoperative Ki67, the prognosis of all negative patients was the best, and that of all positive patients was the worst. Cox regression analysis showed that positive CEA+positive CYFRA21-1+high expression of Ki67 was an independent risk factor for poor prognosis in patients with non-small cell lung cancer (P<0.05). Conclusion The combined detection of preoperative serum CYFRA21-1, CEA, and postoperative Ki67 has important value in evaluating the prognosis of non-small cell lung cancer patients.
6.Bibliometrics-based visualization analysis of xenotransplantation(2014-2023)
Shujun YANG ; Hao WEI ; Jiang PENG ; Panfeng SHANG ; Shengkun SUN
Organ Transplantation 2024;15(5):816-823
Objective To analyze the research progress and development trend of xenotransplantation in China.Methods Literatures related to xenotransplantation in Chinese and English were searched from CNKI and Web of Science Core Collection databases from January 1,2014 to December 31,2023.Visualization analysis of the research progress,hot topics and research frontiers in this field was performed by CiteSpace software and R language from the aspects of number of publications,keywords,distribution of journal origin,core author cooperation network,main research institutions,and citation of key literatures,etc.Results Chinese scholars published 684 articles in Chinese and 624 articles in English,ranking 2nd worldwide.The top 3 Chinese journals regarding the number of publications were Organ Transplantation,Practical Journal of Organ Transplantation(Electronic Version)and Acta Laboratorium Animalis Scientia Sinica.The top 3 English journals were Xenotransplantation,Frontiers in Immunology and Scientific Reports.The top 5 authors regarding the number of publications in Chinese were Pan Dengke,Cai Zhiming,Mou Lisha,Li Xiao and Dou Kefeng.The top 5 authors regarding the number of publications in English were Mou Lisha,Cai Zhiming,Dai Yifan,Wang Yi and Pan Dengke.High-frequency Chinese keywords included xenotransplantation,immune rejection,transplantation,xenogenic,animal model,xenogeneic bone,liver transplantation,gene editing,kidney transplantation,burn and bone transplantation.High-frequency English keywords were expression,xenotransplantation,cells,transplantation and survival.Conclusions In recent years,certain research achievement and global influence have been obtained in the field of xenotransplantation in China,involving a variety of disciplines and a wide range of research contents.However,cooperation among different institutions is still lacking.In the future,cross-regional and interdisciplinary communication and cooperation should be prompted.Resource sharing and integration should be strengthened to maintain rapid and steady advancement in this field.
7.A systematic review of the current nomenclature and classification of congenital auricular malformations
Wenkang LUAN ; Shujun FAN ; Dongwen JIANG ; Chenhao MA ; Leren HE
Chinese Journal of Plastic Surgery 2024;40(10):1127-1136
The nomenclature of congenital auricular malformations is complex and confusing, which brings great inconvenience to clinical work and scientific research. This study comprehensively summarized and sorted out the existing Chinese and English names of congenital auricular malformations, providing reference basis for the scientific classification and clinical treatment of congenital auricular malformations. Sixty-seven papers on the topic of congenital auricle malformations were collected from databases such as CNKI, Wanfang Database, VIP Database, PubMed, Web of Science, and ScienceDirect by using the PRISMA method. The two core issues of morphological and anatomical naming and classification of congenital auricle malformations were sorted and summarized based on these articles. In this study, the existing Chinese and English names of congenital auricular malformations were summarized, and a preliminary classification was made at the morphological and anatomical levels, and the embryological origin of the external auricle and its substructures was elaborated. Based on the result of this study, a comprehensive analysis can be conducted on the malformations of the ear structures and their embryological origins in order to explore the etiology of congenital ear malformations. It is expected that scientific and standardized naming and classification method can be developed, and reasonable and effective standardized treatment can be formulated for congenital auricular malformations based on this study.
8.A systematic review of the current nomenclature and classification of congenital auricular malformations
Wenkang LUAN ; Shujun FAN ; Dongwen JIANG ; Chenhao MA ; Leren HE
Chinese Journal of Plastic Surgery 2024;40(10):1127-1136
The nomenclature of congenital auricular malformations is complex and confusing, which brings great inconvenience to clinical work and scientific research. This study comprehensively summarized and sorted out the existing Chinese and English names of congenital auricular malformations, providing reference basis for the scientific classification and clinical treatment of congenital auricular malformations. Sixty-seven papers on the topic of congenital auricle malformations were collected from databases such as CNKI, Wanfang Database, VIP Database, PubMed, Web of Science, and ScienceDirect by using the PRISMA method. The two core issues of morphological and anatomical naming and classification of congenital auricle malformations were sorted and summarized based on these articles. In this study, the existing Chinese and English names of congenital auricular malformations were summarized, and a preliminary classification was made at the morphological and anatomical levels, and the embryological origin of the external auricle and its substructures was elaborated. Based on the result of this study, a comprehensive analysis can be conducted on the malformations of the ear structures and their embryological origins in order to explore the etiology of congenital ear malformations. It is expected that scientific and standardized naming and classification method can be developed, and reasonable and effective standardized treatment can be formulated for congenital auricular malformations based on this study.
9.Preliminary report of perioperative monitoring of six-gene-edited pig-to-cynomolgus monkey kidney xenotransplantation
Shujun YANG ; Hao WEI ; Yong XU ; Heng'en WANG ; Xiangyu SONG ; Zhibo JIA ; Jiang PENG ; Mengyi CUI ; Boyao YANG ; Leijia CHEN ; Aitao GUO ; Xiaoli ZHANG ; Dengke PAN ; Jiaxiang DU ; Panfeng SHANG ; Shengkun SUN
Organ Transplantation 2023;14(4):521-
Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K+ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β2-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week.
10.Spatio-temporal aggregation of hemorrhagic fever with renal syndrome in Shandong Province from 2017 to 2020
Qing DUAN ; Yufang XING ; Zengqiang KOU ; Xiaomei ZHANG ; Bo PANG ; Xueying TIAN ; Yuwei ZHANG ; Wenji ZHAI ; Zhiqiang WANG ; Xiaolin JIANG ; Shujun DING
Chinese Journal of Endemiology 2022;41(9):715-721
Objective:To study the epidemiological characteristics and spatio-temporal aggregation of hemorrhagic fever with renal syndrome (HFRS) in Shandong Province, and to provide reference for formulating reasonable prevention and control strategies.Methods:Retrospective analysis was used to collect HFRS surveillance data and confirmed case data in Shandong Province from 2017 to 2020 in the "China Disease Prevention and Control Information System Infectious Disease Surveillance System". Geoda 1.18 software was used for global and local spatial autocorrelation analysis, SaTScan 9.6 software was used for spatio-temporal scanning analysis, and ArcGis 10.7 software was used for map drawing and visual display.Results:A total of 3 753 cases of HFRS were reported in Shandong Province from 2017 to 2020, including 56 deaths. The annual incidence rate was 1.26/100 000, 1.22/100 000, 0.75/100 000 and 0.53/100 000, respectively, with an average annual incidence rate of 0.94/100 000. The incidence of HFRS was obviously seasonal, mainly concentrated in autumn and winter from October to December, accounting for 50.41% (1 892/3 753). The age of onset was mainly 30-59 years old, accounting for 61.68% (2 315/3 753). The male to female ratio was 2.76 ∶ 1.00 (2 756 ∶ 997). The occupation distribution was mainly farmers, accounting for 81.99% (3 077/3 753). The global spatial autocorrelation analysis showed that HFRS showed spatial aggregation areas in each year from 2017 to 2020 (Moran' I = 0.38, 0.33, 0.59, 0.46, Z = 7.47, 7.23, 10.69, 8.66, P < 0.001). The local spatial autocorrelation analysis showed that "high-high" aggregation areas were mainly concentrated in central and southeast of Shandong Province, while "low-low" aggregation areas were mainly concentrated in northwest of Shandong Province. Spatio-temporal scanning analysis revealed 1 type Ⅰ agglomerations and 2 type Ⅱ aggregation areas. The type Ⅰ aggregation areas occurred from October to November 2018, covering 22 counties (districts) of 5 cities in Qingdao, Yantai, Weifang, Weihai and Rizhao. The first type Ⅱ aggregation area occurred from October to November 2017, involving 23 counties (districts) of 8 cities in Jinan, Zibo, Zaozhuang, Weifang, Jining, Tai 'an, Rizhao and Linyi. The second type Ⅱ aggregation area occurred in Jinxiang County, Jining City from February to March 2017. Conclusion:The incidence of HFRS in Shandong Province from 2017 to 2020 has obvious spatio-temporal aggregation, and the hot spots are concentrated in central and southeast of Shandong Province, which should be regarded as a key area for prevention and control of HFRS.

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