1.TBX1 inhibits the activation of MAPK and PI3K signaling pathway and reduces the proliferation of colorectal cancer cells thorough activating PARK2
Jingni ZHANG ; Tong HUANG ; Mengge BAI ; Jianling WANG ; Man ZHAO
Cancer Research and Clinic 2024;36(11):808-816
Objective:To investigate the effect of the transcription factor TBX1 on the proliferation of colorectal cancer cells and to explore potential molecular mechanisms.Methods:The mRNA and protein levels of TBX1 in colorectal cancer cell lines HCT116, RKO, SW480, HT29, and LOVO were detected by using reverse transcription quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Colorectal cancer cell lines HCT116 and SW480 cells with low TBX1 expression were transfected with either a pcDNA3.1 plasmid containing TBX1 mimics (TBX1 overexpression group) or an empty pcDNA3.1 plasmid (the control group). LOVO cells with high TBX1 expression were transfected with small interfering RNA (siRNA) targeting TBX1 including si-TBX1-8604A, si-TBX1-8604B, and a negative control siRNA (si-NC), which were treated as si-TBX1-8604A group, si-TBX1-8604B group, and si-NC group. qRT-PCR was used to detect the expressions of transcriptional level TBX1 and PARK2, and Western blot was used to detect the protein levels of TBX1, PARK2, and key factors in the MAPK and PI3K signaling pathways. Methyl Thiazolyl Tetrazolium (MTT) assay and cell colony formation assay were used to detect the cell proliferation. Combining literatures and the JASPAR database, 2 binding sites of TBX1 in the PARK2 promoter region were predicted. Chromatin immunoprecipitation assay was employed to verify the binding sites of TBX1 to PARK2 in HCT116 and SW480 cells. Dual luciferase reporter gene assay was used to verify the targeting relationship between TBX1 and PARK2. The expression of TBX1 and PARK2 in colon cancer tissues was analyzed by using the Cancer Genome Atlas (TCGA) database (September 2023).Results:High TBX1 expression in HCT116 and SW480 cells transfected with TBX1 mimics plasmid was confirmed by qRT-PCR and Western blot, while TBX1 expression was successfully knocked down in LOVO cells transfected with siRNA targeting TBX1. MTT assay indicated that the absorbance values for HCT116 cells in TBX1 overexpression group on d1, d3, d5, and d7 after inoculation, and for SW480 cells on d3, d5, and d7 after inoculation were lower than those in the control group, and the differences were statistically significant (all P < 0.01). LOVO cells in the si-TBX1-8604A group and si-TBX1-8604B group exhibited higher absorbance values than the si-NC group on d1, d3, d5, and d7 after inoculation, and the differences were statistically significant (all P < 0.05). Cell colony formation assay revealed that after 14 d, the colony number of HCT116 cells [(387±9) vs. (843±13)] and SW480 cells [(413±9) vs. (931±15)] in TBX1 overexpression group was lower than that in the control group, and the differences were statistically significant (all P < 0.05). The colony number of LOVO cells in the si-TBX1-8604A group and si-TBX1-8604B group was (493±77) and (470±32), respectively, which was higher than that in the si-NC group (349±26), and the differences were statistically significant (all P < 0.05). The protein relative expression levels of p-ERK and p-AKT S473 in HCT116 and SW480 cells in TBX1 overexpression group were lower than those in the control group, while protein relative expression levels of p-ERK and p-AKT S473 in LOVO cells in the si-TBX1-8604A group and si-TBX1-8604B group were higher than those in the si-NC group, and the differences were statistically significant (all P < 0.05). The relative expression level of PARK2 mRNA in HCT116 and SW480 cells (all P < 0.01) and the protein level in the overexpression group were higher than those in the control group. Chromatin immunoprecipitation assay demonstrated that the enrichment times of TBX1 binding to 2 sites of PARK2 intron in HCT116 and SW480 cells in TBX1 overexpression group were higher than that in the control group, and the differences were statistically significant (all P < 0.05). Dual luciferase reporter gene assay showed that the relative luciferase activity of HCT116 and SW480 cells co-transfected with pcDNA3.1 plasmid containing TBX1 mimics and pGL3 plasmid containing PARK2 mimics was higher than that of cells co-transfected with empty pcDNA3.1 and pGL3 plasmids, co-transfected with empty pcDNA3.1 plasmid and pGL3 plasmid containing PARK2 mimics, co-transfected with pcDNA3.1 plasmid containing TBX1 mimics and empty pGL3 plasmid, and the differences were statistically significant (all P < 0.05). Spearman analysis showed that there was a positive correlation between transcriptional level TBX1 and PARK2 in colon cancer tissues (288 cases) in TCGA database ( r = 0.226, P < 0.001); and the relative expression level of PARK2 mRNA in colon cancer tissues (383 cases) was lower than that in normal intestinal tissues (50 cases), and the difference was statistically significant ( P < 0.001). Conclusions:Elevated expression of transcriptional factor TBX1 inhibits the proliferation of colorectal cancer cells, potentially by activating the downstream target gene PARK2 and inhibiting the phosphorylation of ERK and AKT in the MAPK and PI3K signaling pathways, ultimately affecting the activation of these pathways.
2.Quantitative Evaluation of Elderly Health Policies in China Based on PMC Index Model
Anli WANG ; Jianling WANG ; Wenting ZHANG ; Haibo LUO ; Tingxiao WEN ; Feilong YANG
Journal of Medical Informatics 2024;45(11):36-42
Purpose/Significance The paper discusses the construction of the current elderly health policy system by using policy text min-ing and quantitative evaluation,and provides references for future policy adjustment and optimization.Method/Process Policies on elderly health issued by the relative departments from 2019 to 2023 are analyzed.The ROST text data mining tool is used to extract high-frequency words and build semantic networks.Gephi is utilized to visualize social network relationships for text analysis.Quantitative analysis of these policies is conducted using the PMC index model.Results/Conclusion The high-frequency words such as"national","society","knowl-edge","resources",and"guarantee"are identified as key areas in elderly health policies,which are interconnected within the social network.The overall trend of the elderly health policy formulation is positive.It is suggested to improve the accuracy and adaptability of the policy,clari-fy the time node and core elements of the policy,and enhance the strategic analysis of the function and content of the policy.
3.Teprenone alleviates LPS-induced inflammatory response and cardiac dysfunction through E3 ubiquitin ligase CHIP
Liting XU ; Yingwen LIU ; Jianling LI ; Wan LIN ; Miao WANG ; Lei YU ; Xue ZHANG ; Hang LI ; Huadong WANG ; Xiuxiu LÜ ; Yiyang WANG
Chinese Journal of Pathophysiology 2024;40(5):862-871
AIM:To explore the therapeutic effect of teprenone(geranylgeranylacetone,GGA)on lipopolysac-charide(LPS)-induced cardiac dysfunction and its mechanism.METHODS:(1)Eight-week-old male C57BL/6 wild-type mice and carboxyl terminus of heat shock protein 70(HSP70)-interacting protein(CHIP)gene knockout mice were randomly divided into control group,LPS group,LPS+GGA group and GGA group,with 8 mice in each group.The model was established by intraperitoneal injection of LPS(25 mg/kg),and 1 h after LPS stimulation,mice were given intraperito-neal injection of GGA(100 mg/kg).The technique of high-resolution ultrasonography system was used to evaluate the car-diac function of mice.The serum of mice from each group were collected to detect the levels of creatine kinase-MB(CK-MB)and lactate dehydrogenase(LDH).HE staining was performed to observe histological changes of cardiac tissues.ELISA was used to detect the levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)in cardiac tissues.West-ern blot was used to detect the protein levels of HSP70,CHIP,karyopherin-α 2(KPNA2),myeloperoxidase(MPO),vas-cular cell adhesion molecule(VCAM),intercellular cell adhesion molecule(ICAM),and nuclear factor-κB(NF-κB)in cardiac tissues.(2)In vitro cell inflammation model was established using mouse myocardial cells HL-1 stimulated with LPS.ELISA was used to detect the levels of TNF-α and IL-6 in cell supernatants.Western blot was used to detect the pro-tein expression levels of HSP70,CHIP,and KPNA2 in myocardial cells.Immunofluorescence staining was performed to observe the content of nuclear NF-κB.RESULTS:(1)GGA effectively improved cardiac function of LPS-stimulated mice,significantly increased ejection fraction and left ventricular fractional shortening(P<0.01),reduced serum levels of CK-MB and LDH(P<0.01),and alleviated myocardial injury.(2)GGA significantly reduced the release of TNF-α and IL-6 caused by LPS(P<0.01),as well as nuclear translocation of NF-κB,decreased the levels of KPNA2,MPO,VCAM and ICAM in cardiac tissues,and increased the levels of HSP70 in cardiac tissues and cells(P<0.01).(3)In CHIP knockout myocardial cells and mice,GGA failed to inhibit LPS-induced inflammatory response and lost its effect on im-proving cardiac function.CONCLUSION:The protective effect of GGA against LPS-caused cardiac dysfunction of mice is related to increasing expression of HSP70 and promoting CHIP activation,which inhibits the translocation of NF-κB into nucleus and suppresses inflammatory factor release.CHIP knockout abolishes the effects of GGA on reducing LPS-induced inflammatory response and myocardial injury.
4.Research progress on the role of SHP-2 in tumor-associated macrophages
Xueliang WU ; Jianchun FAN ; Fei GUO ; Qi ZHANG ; Jun XUE ; Ximo WANG ; Guangyuan SUN ; Jianling LIU ; Lei HAN ; Shuquan GAO
Chinese Journal of Comparative Medicine 2024;34(1):171-176
Tumor-associated macrophages(TAMs)are the predominant cell group in the tumor microenvironment(TME)and are the most important regulatory cells of immune system suppression and tumor cell proliferation in TIME.Src homology-2 domain-containing protein tyrosine phosphatase 2(SHP-2)is a non-receptor protein tyrosine phosphatase that plays an important role in the transmission of signals from the cell surface to the nucleus.SHP-2 is a key intracellular regulatory factor mediating cell proliferation and differentiation and is involved in a variety of growth factor and cytokine signaling pathways linking the cell surface to the nucleus.Recent studies have shown that SHP-2 is a key enzyme in determining the function of TAMs,but because of its variable function,it plays different or even opposite roles in different solid TMEs.This paper reviews the function of SHP-2 in TAMs and related solid tumors to provide a comprehensive reference for tumor immunity and targeted therapy research.
5.Fatigue status and influencing factors of heavy-duty commercial truck drivers
Jianjiang ZHANG ; Shaoying WANG ; Siping CHEN ; Jianling LIU
Journal of Environmental and Occupational Medicine 2024;41(7):789-795
Background Fatigue driving is an important cause of road traffic accidents in modern society, and the fatigue condition of heavy-duty commercial truck drivers has attracted widespread attention. Research on the fatigue status and influencing factors of heavy-duty commercial truck drivers in China is relatively rare at present. Objective To analyze the main characteristics of fatigue among heavy-duty commercial truck drivers and the impacts of factors such as working hours, insomnia, and occupational burnout on their fatigue status. Methods Using cluster sampling method, a cross-sectional study was conducted from July to August 2023, enrolling heavy-duty commercial truck drivers in long-distance freight logistics markets (stations) located in three administrative regions of W City, Xinjiang Uygur Autonomous Region. A self-designed questionnaire was used to collect demographic and occupational characteristics of heavy-duty commercial truck drivers, and the Chinese versions of Fatigue Scale-14 (FS-14), Athens Insomnia Scale (AIS), and Maslach Burnout Inventory General Survey (MBI-GS) were used to evaluate their fatigue, insomnia, and occupational burnout status, respectively. Mann-Whitney U test and Kruskal-Walls H test were used to compare intergroup differences, and Spearman correlation coefficient was used to analyze the correlation between variables. Hierarchical regression models were used to study the impacts of selected variables on fatigue status. Results This study obtained 311 valid questionnaires, with a valid recovery rate of 88.86% (311/350). The physical fatigue, mental fatigue, and total fatigue scores of the survey subjects in M (P25, P75) were 3.00 (2.00, 4.00), 2.00 (1.00, 3.00), and 5.00 (4.00, 6.00), respectively. The comparison results showed that, except for smoking, there were statistically significant differences in total fatigue scores between different groups of age, marital status, number of children, educational level, service length of freight transportation, average daily working time, and average monthly income (P<0.05). The difference in total fatigue score among the groups without sleep disorders, with suspected insomnia, and with insomnia was statistically significant (P<0.001). The difference in total fatigue score among the groups without occupational burnout, with moderate occupational burnout, and with severe occupational burnout was also statistically significant (P<0.001). Positive correlations were found between insomnia score and scores of physical fatigue (rs=0.507), mental fatigue (rs=0.547), and total fatigue (rs=0.618) (P<0.001). Hierarchical regression models revealed that having more children, extended daily working hours, insomnia, and increased scores of decreased personal accomplishment were negative factors affecting the fatigue status of heavy-duty commercial truck drivers (P<0.05), and the final regression equation was: total fatigue score=7.579+0.581×number of children+0.916×average daily working time+0.434×score of AIS+0.754×score of reduced personal accomplishment. Conclusion The fatigue status of heavy-duty commercial truck drivers is not optimistic. An increase in the number of children, extended daily working hours, severe insomnia symptoms, and increased scores of decreased personal accomplishment associate with their worse fatigue status.
6.The practice of the cultivation strategy of incremental training of modern clinical thinking ability to cultivate clinical post competency of "5 + 3" integration students
Fengming YUE ; Dong YANG ; Shuming LU ; Quan BAO ; Weisheng ZHANG ; Jianling DU ; Zhijun DUAN
Chinese Journal of Medical Education Research 2023;22(2):168-172
At present, some "5+3" integration students have different levels of understanding and application problems in various stages, such as role transformation, professional knowledge and technology, communication ability and humanistic care ability, clinical thinking and evidence-based medicine concepts, clinical research thinking, learning and work attitude. This research will permeate and run through the training of "5+3" integrated students' diagnostic and therapeutic operation ability through the training of modern clinical thinking oriented by post competency, and integrate humanistic care, evidence-based medicine, learning attitude, working attitude, and attitude towards patients in the whole process to gradually complete the comprehensive training goal of clinical thinking oriented by post competency + diagnostic and therapeutic operation ability.
7.Genotyping and traceability analysis of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau by MLVA
Li MA ; Xuefei ZHANG ; Hongmei XUE ; Aiping ZHANG ; Lingling REN ; Teng QI ; Yuanbo ZHAO ; Jianling WANG ; Xuxin YANG ; Jiquan LI
Chinese Journal of Endemiology 2023;42(4):269-273
Objective:To observe multiple locus variable-number tandem repeat analysis (MLVA) typing of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province, and to explore the relationship between the strains and strains previous isolated from Qinghai Province. Methods:Blood samples of Himalayan marmot were collected in Qinghai-Tibet Plateau of Qinghai Province from March 2019 to October 2020. Pathogens were isolated and cultured from Brucella antibody positive samples identified by using the rose bengal test (RBT). Conventional biological methods and molecular biological methods (BCSP31-PCR and AMOS-PCR) were used for strain identification. At the same time, MLVA method was used to genotype the isolated strains, and cluster analysis was used to analyze the genetic relationships between the strains based on the genotype of 70 Brucella isolated from different hosts in Qinghai Province. Results:A total of 1 466 blood samples of Himalayan marmot were collected from Qinghai-Tibet Plateau. Two strains of Brucella were isolated and cultured from 64 RBT-positive samples, named QH2013054 and QH2013062, respectively. They were identified as Brucella ovis biotype Ⅲ by conventional and molecular biological methods. The MLVA genotyping results showed that QH2013054 and QH2013062 were different at the Bru16 locus, indicating different MLVA genotypes. Cluster analysis showed that strain QH2013054 had the same MLVA genotype as 7 strains, among which 6 strains were from 3 farmers and 3 sheep from the same family in Gonghe County, and 1 strain was from a farmer in Menyuan Hui Autonomous County. The strain QH2013062 had the same MLVA genotype as 4 strains, including 3 strains from 3 farmers in Menyuan Hui Autonomous County and 1 strain from a farmer in Tu Autonomous County of Huzhu. Conclusions:The strains of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province have the same MLVA genotype as some strains of Brucella isolated from humans and sheep in Qinghai Province. It is speculated that the host humans, sheep and Himalayan marmot in Qinghai-Tibet Plateau may have a common source of infection.
8.Rehabilitation exercise attitudes and compliance of patients undergoing coronary artery bypass grafting and their influencing factors
Jing ZHANG ; Jiashuai ZHANG ; Jianling DING ; Jianshuang TAO ; Xiuxiu TAO ; Yanli BEN
Chinese Journal of Modern Nursing 2023;29(12):1569-1575
Objective:To explore the status quo of exercise rehabilitation attitudes and compliance in patients undergoing coronary artery bypass grafting (CABG), and analyze the influencing factors, so as to provide evidence for improving the compliance of rehabilitation exercise in CABG patients.Methods:From January to August 2022, convenience sampling was used to select 228 patients with CABG in three ClassⅢ Grade A hospitals in Urumqi City as the survey subject. One month after surgery, a questionnaire survey was conducted using the General Information and Disease Information Questionnaire, the Questionnaire of Rehabilitation Exercises (QRE), the Chinese version of the Tampa Scale of Kinesiophobia for the Heart (TSK Heart), and the Chinese version of the Multidimensional Fatigue Symptom Inventory-Short Form (MFSI-SF-C). This study conducted a latent category analysis of rehabilitation exercise attitudes and compliance among CABG patients, and identified the influencing factors of their latent categories through Logistic regression. A total of 220 valid questionnaires were collected, with an effective recovery rate of 96.49% (220/228) .Results:The fitting results of latent category analysis showed that the rehabilitation exercise attitudes and compliance of 220 CABG patients could be divided into 4 latent categories, which were respectively named as: excellent attitude-excellent compliance group, good attitude-single exercise group, moderate attitude-poor compliance group, poor attitude-poor compliance group. Logistic regression showed that education level, nationality, pain, exercise fear and fatigue were independent influencing factors for the latent categories of exercise rehabilitation attitudes and compliance in CABG patients ( P<0.05) . Conclusions:Postoperative exercise rehabilitation attitudes and compliance in CABG patients have 4 latent categories, and patient education level, nationality, pain, exercise fear and fatigue are all factors that affect compliance. Medical and nursing workers should formulate targeted interventions based on influencing factors to improve the compliance of exercise rehabilitation in patients with CABG.
9.Research progress and enlightenment of evaluation of long-term care service quality at home and abroad
Yanqiu WENG ; Jianling HAO ; Wenyao CHEN ; Lingjuan ZHANG
Chinese Journal of Modern Nursing 2023;29(2):161-164
The aging of the population has led to a sharp increase in the demand of the elderly for long-term care services. How to ensure the quality of long-term care services is a hot topic of widespread concern in society. In developed countries, long-term care services have been carried out early, a perfect service quality evaluation system has been formed, and the evaluation results have been effectively used. Long-term care services in China started late, there is no unified evaluation standard for the quality of long-term care services, and the evaluation indicators are diversified. It is necessary to explore a long-term care service quality evaluation system with Chinese characteristics based on foreign experience, covering the whole process of long-term care services. In addition, core indicators need to be selected to reflect the quality of long-term care services, strengthen the application of evaluation results, improve the effectiveness of long-term care service quality evaluation, and promote the sustainable development of long-term care in China.
10.Construction of evaluation criteria for long-term care ability of the elderly
Jing LIU ; Xiaoying LU ; Jianling HAO ; Wenyao CHEN ; Yanqiu WENG ; Lingjuan ZHANG
Chinese Journal of Modern Nursing 2023;29(2):165-169
Objective:To explore the construction of evaluation criteria for long-term care ability of the elderly suitable for China's national conditions.Methods:The evaluation criteria were initially established through literature research and field semi-structured interviews. Through the Delphi method, 15 experts were consulted and the evaluation criteria for the long-term care ability of the elderly were finally established.Results:The effective rate of the two rounds of expert consultation questionnaires was 100%, and the expert authority coefficients were 0.91 and 0.92 respectively. The evaluation criteria for long-term care ability of the elderly were established, including two first-level indicators of self-care ability and disease severity, four second-level indicators of abilities of daily living, mental state and social participation, perception and communication, and disease severity, as well as 39 third-level indicators including recumbent status, cognitive function, level of consciousness, illness and so on.Conclusions:The evaluation criteria for long-term care ability of the elderly are scientific and reliable, which can provide a reference for the classification of long-term care services for the disabled elderly in China.

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