1.Construction and Functional Validation of GTKO/hCD55 Gene-Edited Xenotransplant Donor Pigs
Jiaoxiang WANG ; Lu ZHANG ; Shuhan CHEN ; Deling JIAO ; Heng ZHAO ; Taiyun WEI ; Jianxiong GUO ; Kaixiang XU ; Hongjiang WEI
Laboratory Animal and Comparative Medicine 2025;45(4):379-392
Objective To develop GTKO (α-1,3-galactosyltransferase gene-knockout, GTKO)/hCD55 (human CD55) gene-edited xenotransplant donor pigs and verify their function. Methods In this study, CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated nuclease 9), PiggyBac transposon technology and somatic cell nuclear transfer technology were used to construct GTKO/hCD55 gene-edited Diannan miniature pigs. The phenotype and function of GTKO/hCD55 pigs were analyzed by Sanger sequencing, real-time fluorescence quantitative PCR, flow cytometry, immunofluorescence, bisulfite sequencing, antigen-antibody binding assays, and complement-dependent cytotoxicity assays. Results After transfection of PX458 and PiggyBac gene editing vectors into wild-type fetal pig fibroblasts, 48 single-cell colonies were obtained through puromycin drug screening. Two single-cell colonies were selected for somatic cell nuclear transfer, resulting in two fetal pigs at 33 days of gestation. The GGTA1(α-1,3-galactosyltransferase) genotypes of fetal pig F01 were -17 bp and wild type (WT), while the GGTA1 genotypes of fetal pig F02 were -26 bp/+2 bp and -3 bp. The hCD55 mRNA expression levels of both fetal pigs were significantly higher than those of WT pigs (P<0.01). The fetal pig F02 was selected as the donor cell source for recloning, 11 surviving piglets were obtained, all identified as GTKO/hCD55 gene-edited pigs. These pigs showed absence of α-Gal antigen expression, but weak or no expression of hCD55 was observed. Methylation analysis of the hCD55 gene's CpG island showed hypermethylation in kidney tissue lacking hCD55 expression, whereas it was not methylated or partially methylated in kidney tissue expressing hCD55. Moreover, codon optimization of the CpG island of the hCD55 gene to reduce CG content could achieve stable expression of the hCD55 gene. In addition, antigen-antibody binding experiment showed that the amount of human IgM binding to GTKO/hCD55 gene-edited pig fibroblasts was significantly lower than that of WT pigs (P<0.01). Complement-dependent cytotoxicity experiment showed that the survival rate of fibroblasts in GTKO/hCD55 pigs was significantly higher than that in WT pigs (P<0.01). Conclusion This study demonstrates the successful generation of GTKO/hCD55 gene-edited xenotransplant donor pigs. Methylation-induced gene silencing of the hCD55 gene can be effectively avoided by reducing the CG content of the CpG island through codon optimization. This study provides a reference for the development of xenotransplant donor pigs and guides subsequent research on xenotransplantation.
2.Location specificity of the acupoints selected by the syndrome differentiation of acupuncture and moxibustion in weight loss: the possibility of "local weight loss" and "general regulation" based on complex network analysis.
Jiahui XIE ; Shuhan ZHANG ; Yufei WANG ; Mingye GU ; Bin XU ; Tiancheng XU
Chinese Acupuncture & Moxibustion 2025;45(11):1681-1688
OBJECTIVE:
To explore the characteristics and rules of the locations of acupoints selected in weight loss after syndrome differentiation of acupuncture and moxibustion, and provide quantitative evidence for the location specificity of acupoint selection in weight loss.
METHODS:
Clinical research articles on acupoint selection based on syndrome differentiation of acupuncture and moxibustion in weight loss were retrieved in China National Knowledge Infrastructure (CNKI), from the inception to September 20th, 2024, and the data about acupoints and differentiated syndromes were extracted. Based on graph theory, the acupoint-syndrome network was established and its topological parameters such as node degree, value of betweenness centrality, description length and number of community were calculated.
RESULTS:
①The description length of the limbs was 4 255.592, and that of the trunk was 3 274.312. The information contained in the acupoint-syndrome network for the acupoints on the limbs was greater than that for those on the trunk. ②The value of betweenness centrality and node degree showed a nonlinear relationship, with R²of 0.812 1 for the trunk and 0.321 8 for the limbs. The values of betweenness centrality for the acupoints on the trunk were uniformly distributed, and the difference among these values was much smaller than that for the acupoints on the limbs. It suggested that the distance from each trunk acupoint to network center was similar, and the importance among these acupoints to network was similar, while the importance among acupoints located on the limbs was different significantly. ③The frequency proportion of acupoints on the trunk showed uniform distribution among different syndromes, while that of some acupoints located on the limbs such as Taichong (LR3), Neiting (ST44) and Taixi (KI3) presented a correlation with the syndromes.
CONCLUSION
In weight loss with acupuncture and moxibustion, the correlation between the limb acupoints and syndromes is more diverse and specific than that between the trunk acupoints and syndromes. The differences in acupoint selection for simple obesity treated with acupuncture and moxibustion are mostly reflected in the acupoints on the four limbs rather than those on the trunk. It provides an approach to acupoint selection for "local weight loss" and "general regulation" in treatment with acupuncture and moxibustion.
Humans
;
Acupuncture Points
;
Moxibustion
;
Weight Loss
;
Acupuncture Therapy
;
Obesity/physiopathology*
3.Expert consensus on visualized tele-round and quality control management based on the improvement of clinical practice ability
Wanhong YIN ; Xiaoting WANG ; Ran ZHOU ; Dawei LIU ; Yan KANG ; Yaoqing TANG ; Xiaochun MA ; Jianguo LI ; Zhenjie HU ; Haitao ZHANG ; Wei HE ; Lixia LIU ; Wenjin CHEN ; Ran ZHU ; Jun WU ; Hongmin ZHANG ; Lina ZHANG ; Wenzhao CHAI ; Shihong ZHU ; Wangbin XU ; Rongqing SUN ; Xiangyou YU ; Tianjiao SONG ; Ying ZHU ; Hong REN ; Ai SHANMU ; Qing ZHANG ; Wei FANG ; Xiuling SHANG ; Liwen LYU ; Shuhan CAI ; Xin DING ; Heng ZHANG ; Guang FENG ; Lipeng ZHANG ; Bo HU ; Dong ZHANG ; Weidong WU ; Feng SHEN ; Xiaojun YANG ; Zhenguo ZENG ; Qibing HUANG ; Xueying ZENG ; Tongjuan ZOU ; Milin PENG ; Yulong YAO ; Mingming CHEN ; Hui LIAN ; Jingmei WANG ; Yong LI ; Feng QU ; Gang YE ; Rongli YANG ; Xiukai CHEN ; Suwei LI ; Juxiang WANG ; Yangong CHAO
Chinese Journal of Internal Medicine 2025;64(2):101-109
Turning to critical illness is a common stage of various diseases and injuries before death. Patients usually have complex health conditions, while the treatment process involves a wide range of content, along with high requirements for doctor′s professionalism and multi-specialty teamwork, as well as a great demand for time-sensitive treatments. However, this is not matched with critical care professionals and the current state of medical care in China. Telemedicine, which shortens the distance of medical professionals and the gap of disease diagnosis and treatments in various regions through electronic information, can effectively solve the current problem. Therefore, there is an urgent need to develop a standardized, high-quality visualization telemedicine round system .Therefore, experts have been organized to search domestic and foreign literature on telemedicine round for critically ill patients and to form this consensus based on clinical experiences so as to further improve the level of critical care treatments in regions.
4.Risk factor and prognosis of critically ill patients infected with Acinetobacter baumanni
Naobei YE ; Pan ZHANG ; Jian REN ; Hongxia WANG ; Xingyu QIN ; Haonan SUN ; Shuhan XU ; Ruiqin ZHANG
International Journal of Laboratory Medicine 2025;46(10):1173-1179,1184
Objective To analyze the risk factors of critically ill patients infected with Acinetobacter bau-mannii(AB)and carbapenem resistant Acinetobacter baumannii(CRAB).Methods From January 2022 to June 2023,the data of Intensive Care Unit(ICU)patients admitted to Second Hospital of Shanxi Medical Uni-versity in Shanxi Province were collected.According to whether they were infected with AB,the patients were divided into an observation group and a control group(98 cases each).The observation group was further di-vided into a carbapenem sensitive Acinetobacter baumannii(CSAB)group(72 cases)and a CRAB group(26 cases).Mann-Whitney U test,chi-square test and other univariate and multivariate binary Logistic regression were used to analyze the risk factors of AB and CRAB infection for critically ill patients.The prognosis was analyzed by Kaplan Meier survival analysis.Results Long stay in ICU,previous use of carbapenem drugs and high Acute Physiology and Health Evaluation(APACHE Ⅱ)score were independent risk factors for AB sus-ceptibility(P<0.05),while the independent risk factors for CRAB susceptibility were invasive ventilation and delayed surgery(P<0.01).In addition,CRAB infection,COVID-19 and shock was risk factors for death in critically ill patients,and invasive ventilation,indwelling drainage tube and operation could reduce the risk of death in critically ill patients(P<0.05).Conclusion ICU stay time,APACHE Ⅱ score,previous use of car-bapenem drugs and invasive ventilation increase the risk of AB and CRAB infection in critically ill patients.In-vasive ventilation,indwelling drainage and early surgery could reduce the risk of death from AB and CRAB in-fection in critically ill patients.
5.Effect of Lianpu Yin on Improvement of Duodenal Microinflammation in FD Rats and Its Mechanism via NLRP3 Activation
Yang ZHANG ; Wenliang LYU ; Shuhan ZHOU ; Ningfeng MAO ; Jiawei HE ; Yi ZHAO ; Zixuan XU ; Linlin LIU ; Xueyan WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(6):1693-1698
Objective To investigate the effect of Lianpu Yin on duodenal microinflammation in rats with functional dyspepsia(FD)by regulating NLRP3 activation.Methods Wistar rats were randomly divided into blank group and model group.FD rats were reconstructed by iodoacetamide method(2%sucrose solution containing 0.1%iodoacetamide),and the model was verified.FD model rats were randomly divided into model group,Lianpu Yin group and Moxapride group by random number expression method.After a period of two weeks of administration,measurements were taken to determine the body mass,three-hour food consumption,as well as the rates of gastric emptying and intestinal propulsion.The pathological structure of duodenal tissue was observed by HE staining.The serum levels of IL-1β and IL-18 were quantified using the enzyme-linked immunosorbent assay(ELISA)method.The expression levels of NLRP3 and Caspase-1 in each group were detected by Western blot.Expression levels of NLRP3 and Caspase-1 proteins were detected by immunofluorescence.Results Compared with the blank group,body weight,food intake at 3 h,gastric emptyand intestinal propulsion rate in model group were significantly decreased(P<0.01),and inflammatory infiltration of duodenum tissue appeared in the model group.Meanwhile,the expressions of NLRP3 and Caspase-1 proteins,as well as the levels of IL-1β and IL-18 in the duodenal tissue of the model group,showed significant increasing(P<0.05).Compared with the model group,rats in the Lianpu Yin and Moxapride groups displayed significant increasing in body weight,gastric emptying rate,and intestinal propulsion rate(P<0.01).Additionally,inflammatory infiltration of duodenum tissue reduced in these groups.Furthermore,NLRP3 and Caspase-1 protein expressions,as well as IL-1β and IL-18 levels,significantly decreased in the Lianpu Yin and Moxapride groups compared to the model group(P<0.05).Conclusion Lianpu Yin can treat FD rats by inhibiting duodenal microinflammation and then restoring gastrointestinal motility,which may be related to the abnormal activation of NLRP3 inflammasome.
6.To Explore the Biological Connotation of Dampness-Heat Syndrome of Spleen and Stomach Based on the Correspondence Between Syndrome and Prescription under the Mode of Combining Disease and Syndrome
Hailin YAN ; Wenliang LYU ; Jing XU ; Shuhan ZHOU ; Qinghua GAO ; Siyi ZHANG ; Hanlin ZHANG ; Lin YU ; Xiaohui XU
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(6):1501-1508
Dampness-heat syndrome of the spleen and stomach refers to the evil dampness-heat invading the spleen and stomach,the abnormal rise and fall of the middle jiaoqi machinery,dampness-heat depression and steaming,the physiological dysfunction of the spleen and stomach,with abdominal distention and distention,nausea and lethargy of the limbs,poor loose stool,red tongue and yellow greasy fur,and slippery pulse as the main symptoms of the syndrome,most common in digestive system diseases,among which chronic gastritis is the first.This paper summarized the research results in the past decade and related fields.In the mode of combination of disease and syndrome,based on the principle of corresponding prescription and syndrome,combined with the etiology and pathogenesis evolution of spleen and stomach damp-heat syndrome,the biological connotation of spleen and stomach damp-heat syndrome was explained from various aspects such as inflammation and immune disorders,gastrointestinal motivity disorders,water and humidity loss,endoplasmic reticulum function,and micro-ecological disorders.Enrich the research of the essence of syndrome.
7.Effect of Lianpu Yin on Improvement of Duodenal Microinflammation in FD Rats and Its Mechanism via NLRP3 Activation
Yang ZHANG ; Wenliang LYU ; Shuhan ZHOU ; Ningfeng MAO ; Jiawei HE ; Yi ZHAO ; Zixuan XU ; Linlin LIU ; Xueyan WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(6):1693-1698
Objective To investigate the effect of Lianpu Yin on duodenal microinflammation in rats with functional dyspepsia(FD)by regulating NLRP3 activation.Methods Wistar rats were randomly divided into blank group and model group.FD rats were reconstructed by iodoacetamide method(2%sucrose solution containing 0.1%iodoacetamide),and the model was verified.FD model rats were randomly divided into model group,Lianpu Yin group and Moxapride group by random number expression method.After a period of two weeks of administration,measurements were taken to determine the body mass,three-hour food consumption,as well as the rates of gastric emptying and intestinal propulsion.The pathological structure of duodenal tissue was observed by HE staining.The serum levels of IL-1β and IL-18 were quantified using the enzyme-linked immunosorbent assay(ELISA)method.The expression levels of NLRP3 and Caspase-1 in each group were detected by Western blot.Expression levels of NLRP3 and Caspase-1 proteins were detected by immunofluorescence.Results Compared with the blank group,body weight,food intake at 3 h,gastric emptyand intestinal propulsion rate in model group were significantly decreased(P<0.01),and inflammatory infiltration of duodenum tissue appeared in the model group.Meanwhile,the expressions of NLRP3 and Caspase-1 proteins,as well as the levels of IL-1β and IL-18 in the duodenal tissue of the model group,showed significant increasing(P<0.05).Compared with the model group,rats in the Lianpu Yin and Moxapride groups displayed significant increasing in body weight,gastric emptying rate,and intestinal propulsion rate(P<0.01).Additionally,inflammatory infiltration of duodenum tissue reduced in these groups.Furthermore,NLRP3 and Caspase-1 protein expressions,as well as IL-1β and IL-18 levels,significantly decreased in the Lianpu Yin and Moxapride groups compared to the model group(P<0.05).Conclusion Lianpu Yin can treat FD rats by inhibiting duodenal microinflammation and then restoring gastrointestinal motility,which may be related to the abnormal activation of NLRP3 inflammasome.
8.To Explore the Biological Connotation of Dampness-Heat Syndrome of Spleen and Stomach Based on the Correspondence Between Syndrome and Prescription under the Mode of Combining Disease and Syndrome
Hailin YAN ; Wenliang LYU ; Jing XU ; Shuhan ZHOU ; Qinghua GAO ; Siyi ZHANG ; Hanlin ZHANG ; Lin YU ; Xiaohui XU
World Science and Technology-Modernization of Traditional Chinese Medicine 2025;27(6):1501-1508
Dampness-heat syndrome of the spleen and stomach refers to the evil dampness-heat invading the spleen and stomach,the abnormal rise and fall of the middle jiaoqi machinery,dampness-heat depression and steaming,the physiological dysfunction of the spleen and stomach,with abdominal distention and distention,nausea and lethargy of the limbs,poor loose stool,red tongue and yellow greasy fur,and slippery pulse as the main symptoms of the syndrome,most common in digestive system diseases,among which chronic gastritis is the first.This paper summarized the research results in the past decade and related fields.In the mode of combination of disease and syndrome,based on the principle of corresponding prescription and syndrome,combined with the etiology and pathogenesis evolution of spleen and stomach damp-heat syndrome,the biological connotation of spleen and stomach damp-heat syndrome was explained from various aspects such as inflammation and immune disorders,gastrointestinal motivity disorders,water and humidity loss,endoplasmic reticulum function,and micro-ecological disorders.Enrich the research of the essence of syndrome.
9.Expert consensus on visualized tele-round and quality control management based on the improvement of clinical practice ability
Wanhong YIN ; Xiaoting WANG ; Ran ZHOU ; Dawei LIU ; Yan KANG ; Yaoqing TANG ; Xiaochun MA ; Jianguo LI ; Zhenjie HU ; Haitao ZHANG ; Wei HE ; Lixia LIU ; Wenjin CHEN ; Ran ZHU ; Jun WU ; Hongmin ZHANG ; Lina ZHANG ; Wenzhao CHAI ; Shihong ZHU ; Wangbin XU ; Rongqing SUN ; Xiangyou YU ; Tianjiao SONG ; Ying ZHU ; Hong REN ; Ai SHANMU ; Qing ZHANG ; Wei FANG ; Xiuling SHANG ; Liwen LYU ; Shuhan CAI ; Xin DING ; Heng ZHANG ; Guang FENG ; Lipeng ZHANG ; Bo HU ; Dong ZHANG ; Weidong WU ; Feng SHEN ; Xiaojun YANG ; Zhenguo ZENG ; Qibing HUANG ; Xueying ZENG ; Tongjuan ZOU ; Milin PENG ; Yulong YAO ; Mingming CHEN ; Hui LIAN ; Jingmei WANG ; Yong LI ; Feng QU ; Gang YE ; Rongli YANG ; Xiukai CHEN ; Suwei LI ; Juxiang WANG ; Yangong CHAO
Chinese Journal of Internal Medicine 2025;64(2):101-109
Turning to critical illness is a common stage of various diseases and injuries before death. Patients usually have complex health conditions, while the treatment process involves a wide range of content, along with high requirements for doctor′s professionalism and multi-specialty teamwork, as well as a great demand for time-sensitive treatments. However, this is not matched with critical care professionals and the current state of medical care in China. Telemedicine, which shortens the distance of medical professionals and the gap of disease diagnosis and treatments in various regions through electronic information, can effectively solve the current problem. Therefore, there is an urgent need to develop a standardized, high-quality visualization telemedicine round system .Therefore, experts have been organized to search domestic and foreign literature on telemedicine round for critically ill patients and to form this consensus based on clinical experiences so as to further improve the level of critical care treatments in regions.
10.Progress in animal models of sick sinus syndrome
Ran SUN ; Guanzhen XU ; Yue LIU ; Yingying SUN ; Shuhan ZHANG ; Huiying BO ; Yantong WU ; Ping HOU
Acta Laboratorium Animalis Scientia Sinica 2024;32(9):1198-1206
Sick sinus syndrome(SSS)refers to damage to the sinoatrial node and its surrounding tissues,which leads to excitation and conduction dysfunction of the sinoatrial node,Resultsing in arrhythmia diseases.A better understanding of the pathogenesis of SSS is required to provide a basis for its treatment,including establishing an animal model that can simulate human sinus node dysfunction.In this paper,we review the animal selection,the principles and method of modeling,and the evaluation method and detection indicators of the models,to provide a basis for further studies of the pathogenesis of SSS.

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