1.Residual cell types,molecular expression profiles and quality assessment of in vitro cultured human thymic slices
Wanqing GUAN ; Guihua LUO ; Jingxuan HAN ; Qun XIANG ; Yunfei AN ; Lu ZHAO ; Jianhong MI ; Zeqing FENG ; Yuzhang WU
Journal of Army Medical University 2025;47(9):893-903
Objective To analyze the composition and function of residual cells in pre-transplantation human thymic slices by single-cell transcriptomics sequencing(scRNA-seq),and established a quality assessment method for thymic slices based on the expression levels of molecular markers in the culture supernatant.Methods The discarded thymus from 18 patients with congenital heart disease undergoing surgical treatment in Department of Cardiothoracic Surgery of Children's Hospital Affiliated to Chongqing Medical University from May 2023 to January 2024 were collected and prepared into thymic slices.After the slices were cultured in vitro for 14 d,scRNA-seq was employed to identify the residual cell types,and gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis was performed to analyze the biological function of the residual cells.Then based on the literature concerning thymic slice culture,the molecular markers indicating thymocyte function were screened out.ELISA was applied to detect the changes in protein levels of molecular markers in the supernatant.Receiver operating characteristic(ROC)curve was plotted and assess the value of the molecular markers in the supernatant in evaluating the quality of thymic slices with area under the curve(AUC).Then,the qualified and unqualified thymic slices determined by our obtained molecular markers were transplanted subcutaneously into male nude mice(6~8 weeks old,weighing 14~17 g),respectively,and the male nude mice without transplantation of the thymic slices served as control group.Flow cytometry and histologic analysis were utilized to observe the immune reconstitution after transplantation.Results ① scRNA-seq identified 11 cell types in thymic slices,dominated with epithelial cells,fibroblasts,and T cells.GO and KEGG enrichment analysis showed that epithelial cells were involved in enrichment entries related to chemotaxis,epithelial cell development,cell matrix adhesion and tight junction;fibroblasts were involved in enrichment entries related to extracellular matrix,epithelial cell proliferation,negative regulation of cell migration,and regulation of actin cytoskeleton;T cells were mainly related to T cell differentiation,regulation of T cell activation,T cell apoptosis,and T cell receptor signaling.② Molecular markers,CCL19,CCL21,CXCL12,CXCL16,IL16 and SELL were identified to indicate thymocyte function.Compared with the levels of the first day,the protein secretions of CCL19,CCL21,CXCL12 and CXCL16 were significantly increased during in vitro culture(P<0.05),while the protein secretions of IL16 and L-selectin(protein form of SELL)were significantly decreased(P<0.05).The combined predictor Pre1 from subset of cytokines(IL16 and L-selectin)had the highest value in the quality assessment of thymic slices after 1 d of culture(AUC=0.883),and the combined predictor Pre2 from subset of cytokines(CCL19,CCL21,CXCL12 and CXCL16)had the highest value in the quality assessment after 14 d of culture(AUC=0.948).③ Transplantation in nude mice indicated that the qualified thymic slices could develop to thymus structure in vivo,and effectively increase the proportion of T cells in peripheral blood(P<0.01),while the unqualified thymic slices could not obtain the reconstitution of T cell development.Conclusion The main residual component cells in thymic slices are epithelial cells,fibroblasts and T cells.IL16 and L-selectin can be used as potential indicators to determine the quality of donor thymic samples.CCL19,CCL21,CXCL12 and CXCL16 can effectively evaluate the quality of thymic slices before transplantation.
2.Simultaneous Analysis of Microcystins,Cylindrospermopsin,Anatoxin,and Nodularin in Lake Water by Liquid Chromatography-Tandem Mass Spectrometry
Guanxiang YUAN ; Qing LUO ; Guihua LIU ; Xiaoyun QIN ; Honghe LIU ; Zhaoying LÜ ; Jie JIANG
Journal of Sichuan University (Medical Sciences) 2025;56(5):1189-1196
Objective To establish a method for simultaneous determination of trace levels of microcystins,cylindrospermopsin,anatoxin,and nodularin in lake water based on liquid chromatography-tandem mass spectrometry(LC-MS/MS).Methods After being adjusted to alkaline conditions and mixed with six internal standards,the water samples were enriched using dual HLB and ENVI-Carb cartridges.The eluates were then evaporated under nitrogen,reconstituted,and subjected to instrumental analysis.Both water and acetonitrile containing 0.1%formic acid were used as mobile phases.An ACQUITY UPLC? BEH C18 column(150 mm×2.1 mm,1.7 μm)was selected to separate the target cyanotoxins.Multiple reaction monitoring was applied for data acquisition,and quantification was accomplished using internal standard methods.Results Within certain concentration ranges,all 14 cyanotoxins examined in the study showed good linearity,with all correlation coefficients greater than 0.998.When the water volume was 100 mL,the limits of detection and quantification for the 14 cyanotoxins were 0.1-0.9 ng/L and 0.3-2.9 ng/L,respectively,and spiked recoveries and relative standard deviations were 81.7%-132.9%and 1.2%-14.9%,respectively.In the 10 lake water samples analyzed,cylindrospermopsin,anatoxin-α,and multiple microcystins were detected.Conclusion The method developed in the study has high-throughput capacity,as well as high sensitivity,accuracy,and reliability.The method can be applied in the simultaneous detection of microcystins,cylindrospermopsin,anatoxin,and nodularin in lake water.
3.Research on the Path of Integrating the Life View of "Human Life is Most Important" in Traditional Chinese Medicine into Medical Ethics Education for Medical Students
Yanhong ZHOU ; Jing OUYANG ; Yingbo LI ; Jing ZHANG ; Ying HOU ; Guihua LUO
Chinese Medical Ethics 2024;35(3):332-337
The life view of "human life is the most important" in Traditional Chinese Medicine (TCM) originates from the people-oriented theory of Chinese traditional culture, the benevolence thought of Confucianism, the rebirth thought of Taoism and the compassion thought of Buddhism. Taking the life view of "human life is the most important" as the starting point, it has formed medical ethics thoughts such as the social responsibility of saving the world, the medical practice purpose of treating all patients equally, the meticulous working attitude, the diligent and tireless learning attitude, and the exploration spirit of widely exploring the medical roots, which still has important educational significance. Therefore, the life view of "human life is the most important" should be integrated into the medical ethics education of medical students, and the medical ethics literacy of medical students should be improved through the following ways: strengthen the vocational ideal education of saving the dying and healing the wounded, and enhance the sense of social responsibility of medical students; consolidate the theme of life first classroom education and improve the professional identity of medical students; enrich and innovate the practice forms of protecting life, pay attention to the practicality of medical ethics education.
4.Clinical application of split liver transplantation: a single center report of 203 cases
Qing YANG ; Shuhong YI ; Binsheng FU ; Tong ZHANG ; Kaining ZENG ; Xiao FENG ; Jia YAO ; Hui TANG ; Hua LI ; Jian ZHANG ; Yingcai ZHANG ; Huimin YI ; Haijin LYU ; Jianrong LIU ; Gangjian LUO ; Mian GE ; Weifeng YAO ; Fangfei REN ; Jinfeng ZHUO ; Hui LUO ; Liping ZHU ; Jie REN ; Yan LYU ; Kexin WANG ; Wei LIU ; Guihua CHEN ; Yang YANG
Chinese Journal of Surgery 2024;62(4):324-330
Objective:To investigate the safety and therapeutic effect of split liver transplantation (SLT) in clinical application.Methods:This is a retrospective case-series study. The clinical data of 203 consecutive SLT, 79 living donor liver transplantation (LDLT) and 1 298 whole liver transplantation (WLT) performed at the Third Affiliated Hospital of Sun Yat-sen University from July 2014 to July 2023 were retrospectively analyzed. Two hundred and three SLT liver grafts were obtained from 109 donors. One hundred and twenty-seven grafts were generated by in vitro splitting and 76 grafts were generated by in vivo splitting. There were 90 adult recipients and 113 pediatric recipients. According to time, SLT patients were divided into two groups: the early SLT group (40 cases, from July 2014 to December 2017) and the mature SLT technology group (163 cases, from January 2018 to July 2023). The survival of each group was analyzed and the main factors affecting the survival rate of SLT were analyzed. The Kaplan-Meier method and Log-rank test were used for survival analysis.Results:The cumulative survival rates at 1-, 3-, and 5-year were 74.58%, 71.47%, and 71.47% in the early SLT group, and 88.03%, 87.23%, and 87.23% in the mature SLT group, respectively. Survival rates in the mature SLT group were significantly higher than those in the early SLT group ( χ2=5.560, P=0.018). The cumulative survival rates at 1-, 3- and 5-year were 93.41%, 93.41%, 89.95% in the LDLT group and 87.38%, 81.98%, 77.04% in the WLT group, respectively. There was no significant difference among the mature SLT group, the LDLT group and the WLT group ( χ2=4.016, P=0.134). Abdominal hemorrhage, infection, primary liver graft nonfunction,and portal vein thrombosis were the main causes of early postoperative death. Conclusion:SLT can achieve results comparable to those of WLT and LDLT in mature technology liver transplant centers, but it needs to go through a certain time learning curve.
5.Immune reconstitution efficacy of human thymic slices transplanted to different parts of nude mice
Wanqing GUAN ; Guihua LUO ; Jingxuan HAN ; Jianhong MI ; Zeqing FENG ; Yuzhang WU
Immunological Journal 2024;40(2):113-121
This study was designed to explore the immune reconstitution efficacy of human thymic slices transplanted into renal capsule,subcutaneous or muscle of nude mice,and further explore the optimal location of heterotopic transplantation.The thymus tissue discarded from congenital heart disease patients was made into 0.5-1 mm thick tissue sections and cultured in vitro to remove immune cells.H&E staining and immunohistochemical staining were used to assess the residual tissue structure and cell types in thymic slices,while quantitative PCR methods were used to assess the function of residual cells in thymic slices.Then thymic slices were transplanted into the renal capsule,subcutaneous or muscle of nude mice,and the immune reconstitution efficacy was compared by flow cytometry and histology.Data showed that after 14 days of culture in vitro,the clearance rate of T lymphocytes in the thymic slices was more than 90%,and the epithelial cell network structure of the tissue was intact,while a large number of macrophages,dendritic cells and endothelial cells remained.Quantitative PCR results showed that the gene expression levels of epithelial cell markers and secreted cytokines in cultured thymic slices could be effectively maintained.Flow cytometry showed that at 16 weeks after transplantation,the proportion of T cells in peripheral blood of mice in different transplantation groups were significantly increased,whereas the proportion of T cells in muscle group was the highest.In situ histological examination showed that the regeneration of thymus tissue was detected at all three transplant sites.In addition,the graft detection rate was 40%in the renal capsule group,60%in the subcutaneous group and 100%in the musclegroup.In conclusion,the human thymic slices cultured in vitro for 14 days retain a complete thymic matrix microenvironment.Transplantation of human thymic slices can effectively reconstruct the ratio of T cells in nude mice,and the muscle is the most effective transplantation site.
6.Clinical application of split liver transplantation: a single center report of 203 cases
Qing YANG ; Shuhong YI ; Binsheng FU ; Tong ZHANG ; Kaining ZENG ; Xiao FENG ; Jia YAO ; Hui TANG ; Hua LI ; Jian ZHANG ; Yingcai ZHANG ; Huimin YI ; Haijin LYU ; Jianrong LIU ; Gangjian LUO ; Mian GE ; Weifeng YAO ; Fangfei REN ; Jinfeng ZHUO ; Hui LUO ; Liping ZHU ; Jie REN ; Yan LYU ; Kexin WANG ; Wei LIU ; Guihua CHEN ; Yang YANG
Chinese Journal of Surgery 2024;62(4):324-330
Objective:To investigate the safety and therapeutic effect of split liver transplantation (SLT) in clinical application.Methods:This is a retrospective case-series study. The clinical data of 203 consecutive SLT, 79 living donor liver transplantation (LDLT) and 1 298 whole liver transplantation (WLT) performed at the Third Affiliated Hospital of Sun Yat-sen University from July 2014 to July 2023 were retrospectively analyzed. Two hundred and three SLT liver grafts were obtained from 109 donors. One hundred and twenty-seven grafts were generated by in vitro splitting and 76 grafts were generated by in vivo splitting. There were 90 adult recipients and 113 pediatric recipients. According to time, SLT patients were divided into two groups: the early SLT group (40 cases, from July 2014 to December 2017) and the mature SLT technology group (163 cases, from January 2018 to July 2023). The survival of each group was analyzed and the main factors affecting the survival rate of SLT were analyzed. The Kaplan-Meier method and Log-rank test were used for survival analysis.Results:The cumulative survival rates at 1-, 3-, and 5-year were 74.58%, 71.47%, and 71.47% in the early SLT group, and 88.03%, 87.23%, and 87.23% in the mature SLT group, respectively. Survival rates in the mature SLT group were significantly higher than those in the early SLT group ( χ2=5.560, P=0.018). The cumulative survival rates at 1-, 3- and 5-year were 93.41%, 93.41%, 89.95% in the LDLT group and 87.38%, 81.98%, 77.04% in the WLT group, respectively. There was no significant difference among the mature SLT group, the LDLT group and the WLT group ( χ2=4.016, P=0.134). Abdominal hemorrhage, infection, primary liver graft nonfunction,and portal vein thrombosis were the main causes of early postoperative death. Conclusion:SLT can achieve results comparable to those of WLT and LDLT in mature technology liver transplant centers, but it needs to go through a certain time learning curve.
7.Analysis of the current state of physical activity levels and influencing factors in lung cancer patients a month post-surgery
Qiaoqiao MA ; Jing LUO ; Guihua HAO ; Huxing CAO ; Ting WANG ; Shajing FAN ; Min TANG ; Cuiwen TIAN ; Lili HOU
Chinese Journal of Nursing 2024;59(16):1979-1987
Objective To investigate the current postoperative physical activity levels of lung cancer patients after a month and to analyze the influencing factors.Methods From January to December 2023,a total of 1,195 lung cancer patients from 16 hospitals in eastem,central and westem China were selected as the survey subjects by the convenient sampling method.A cross-sectional survey was conducted by General Information Questionnaires,Health-Related Physical Fitness Surveys,MD Anderson Symptom Inventory,Hospital Anxiety and Depression Scale,Social Support Rating Scale,Self-Rated Abilities for Health Practices Scale,and the International Physical Activity Questionnaire.Results 36 questionnaires with missing data exceeding 10%and 13 duplicate questionnaires were excluded,resulting in the analysis of 1,146 questionnaires.After a month postoperatively,lung cancer patients with moderate and low levels of physical activity were 732(63.9%)and 204(17.8%),respectively.Multivariate regression analysis showed that the influencing factors included residence,average monthly family income,regional lymph node staging,family history of lung cancer,grip strength,hemoglobin,and depression score(P<0.05).Conclusion The physical activity level of lung cancer patients a month after surgery is influenced by a combination of sociodemographic,disease-related,health-related physical fitness,and psychological factors.Clinical healthcare professionals should accurately assess the physical activity levels of lung cancer patients,identify those with insufficient physical activity,and develop and implement personalized intervention strategies based on their physical condition and potential influencing factors to enhance their physical activity levels and improve their quality of life.
8.Application progress of artificial intelligence in the study of speech disorders in Parkinson′s disease
Zhaoxia WEI ; Lihua LI ; Qinglu LUO ; Guihua TANG
Chinese Journal of Neurology 2024;57(11):1259-1263
Parkinson′s disease (PD) is a neurodegenerative disorder that significantly impacts speech and voice, leading to hypokinetic dysarthria, a motor speech disorder. The advent of artificial intelligence (AI) has opened new avenues for the assessment, diagnosis, and rehabilitation of PD-associated speech impairments. This review explores the application of AI in the study of PD speech disorders, focusing on automated speech analysis, machine learning algorithms, and the development of speech pathology databases. This review also discusses the methodologies and technologies employed, such as speech signal processing, feature extraction techniques, classification algorithms, and the symptoms they can detect, including voice quality, articulation, speech rate, and prosody. The role of AI in early diagnosis, disease progression monitoring, treatment evaluation, and remote rehabilitation is highlighted. The review concludes with a discussion on the potential and challenges of AI in this field and recommendations for future research.
9.Research on the Path of Integrating the Life View of "Human Life is Most Important" in Traditional Chinese Medicine into Medical Ethics Education for Medical Students
Yanhong ZHOU ; Jing OUYANG ; Yingbo LI ; Jing ZHANG ; Ying HOU ; Guihua LUO
Chinese Medical Ethics 2022;35(3):332-337
The life view of "human life is the most important" in Traditional Chinese Medicine (TCM) originates from the people-oriented theory of Chinese traditional culture, the benevolence thought of Confucianism, the rebirth thought of Taoism and the compassion thought of Buddhism. Taking the life view of "human life is the most important" as the starting point, it has formed medical ethics thoughts such as the social responsibility of saving the world, the medical practice purpose of treating all patients equally, the meticulous working attitude, the diligent and tireless learning attitude, and the exploration spirit of widely exploring the medical roots, which still has important educational significance. Therefore, the life view of "human life is the most important" should be integrated into the medical ethics education of medical students, and the medical ethics literacy of medical students should be improved through the following ways: strengthen the vocational ideal education of saving the dying and healing the wounded, and enhance the sense of social responsibility of medical students; consolidate the theme of life first classroom education and improve the professional identity of medical students; enrich and innovate the practice forms of protecting life, pay attention to the practicality of medical ethics education.
10. Characteristics of patients visiting emergency department by different ways of transportation
Dong LUO ; Tao CHENG ; Mingming DONG ; Fang SHEN ; Guihua YANG ; Yu CAO ; Rong YAO
Chinese Journal of General Practitioners 2019;18(9):847-850
Objective:
To explore the characteristics of patients visiting emergency department by different ways of transportation.
Methods:
The medical information of 23 039 patients visiting emergency department from January 2017 to December 2017 was obtained through the Hospital Information System (HIS). There were 20 856 patients visiting emergency arranged by themselves (self-visiting group), and 2 183 patients sent by ambulance (ambulance group). The characteristics of two groups were compared.
Results:
The proportion of young patients in the self-visit group was the highest (45.97%, 9 587/20 856), and the proportion of elderly patients in the ambulance group was the highest (40.04%, 874/2 183). Throughout the year, the number of patients who came to the hospital by themselves was the highest in the summer, that by ambulance was the highest in the winter, while that was the lowest in spring for both groups. There was no significant fluctuation in the number of patients who came to the hospital during the week. In the self-visited group, the daily peak hours were 18:00-02:00 at night and the ambulance group were 10:00-18:00. The number of visits for top 5 system diseases were trauma, digestive system, other diseases, respiratory system and urinary system diseases in self-visiting group; and were nervous system, trauma, digestive system, respiratory system and circulatory system diseases in ambulance group. The hospitalization rates of self-visiting group and ambulance group were 14.41% (3 006/20 856) and 76.64% (1 564/2 183), respectively.
Conclusion
The emergency patients with different visiting modes have different characteristics. The information would be useful for rationally allocate emergency resources, optimize the treatment process, improve the emergency capacity and ensure the safety of patients.

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