1.Effect of Serum Containing Zhenwutang on Apoptosis of Myocardial Mast Cells and Mitochondrial Autophagy
Wei TANG ; Meiqun ZHENG ; Xiaolin WANG ; Zhiyong CHEN ; Chi CHE ; Zongqiong LU ; Jiashuai GUO ; Xiaomei ZOU ; Lili XU ; Lin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):11-21
ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ (AngⅡ) and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment, AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h, and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 (CCK-8) to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of B-type natriuretic peptide (BNP) in cell culture supernatant, and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently, the experiment was divided into a blank group (20% blank serum), a model group (20% blank serum + 5×10-5 mol·L-1 AngⅡ), low-, medium-, and high-dose (5%, 10% and 20%) serum containing Zhenwutang groups, an autophagy inhibitor group (1×10-4 mol·L-1 3-MA), and autophagy inducer group (1×10-7 mol·L-1 rapamycin). The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe (Lyso Tracker) and mitochondrial probe (Mito Tracker) co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax), and cytochrome C (Cyt C) in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase (Parkin), phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1), and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3, cleaved Caspase-9, Bax, Bcl-2, and Cyt C in apoptosis-related pathways, phosphorylated ubiquitin ligase (p-Parkin), phosphorylated PTEN-induced kinase 1 (p-PINK1), p62, and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways, and the change of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10-5 mol·L-1, the cell activity was the lowest, and there was no cytotoxicity. At this concentration, the surface area of cardiomyocytes was significantly increased (P<0.01), and the content of BNP in the supernatant of culture medium was significantly increased (P<0.05). Therefore, AngⅡ with a concentration of 5×10-5 mol·L-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group, the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate (P<0.01) and significantly decreased mitochondrial membrane potential (P<0.01). The results of immunofluorescence co-localization showed that compared with the blank group, the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group, the relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 in the model group was significantly up-regulated (P<0.01), while the relative mRNA expression of Bcl-2, Parkin, and PINK1 was significantly down-regulated (P<0.01). In addition, the relative protein expression of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 was significantly up-regulated (P<0.01). The LC3Ⅱ/Ⅰ was significantly decreased, and the relative protein expression of Bcl-2, p-Parkin, p-PINK1, and Beclin1 was significantly down-regulated (P<0.01). Compared with the model group, the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate (P<0.01), and the decrease ratio of mitochondrial membrane potential is significantly lowered (P<0.01) in a dose-dependent manner. Additionally, both red and green fluorescence spots became more in these groups. In the 3-MA group, the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 was significantly down-regulated (P<0.05, P<0.01), while that of Bcl-2, Parkin, and PINK1 was significantly up-regulated (P<0.01). In the serum containing Zhenwutang groups, the relative protein expression levels of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 were significantly down-regulated (P<0.05,P<0.01). The LC3Ⅱ/Ⅰ was significantly increased, and the relative protein expression levels of Bcl-2, p-Parkin, p-PINK1, and Beclin1 were significantly up-regulated (P<0.01). ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.
2.Effect Evaluation of Multidisciplinary Collaborative Diagnosis and Treatment Model for Children with Brain Injury
Xiongwu YU ; Yunli ZHOU ; Zhiyong DING ; Chaohong WANG ; Zeyi XIE ; Hongna LU ; Hua JIN
Journal of Kunming Medical University 2024;45(1):156-162
Objective To summarize the experience of multi-disciplinary team(MDT)in the pediatric department of Qujing Maternal and Child Health Hospital,and to evaluate the effectiveness of MDT on neonatal brain injury.Methods The clinical data of children with brain injury and treated in the pediatrics department of Qujing Maternal and Child Health Hospital from November 2019 to April 2023 were collected.The children with brain injury and treated from October 2019 to June 2020 were regarded as the non-MDT group,and the children with brain injury and treated from July 2020 to April 2023 were regarded as the MDT group for comparative analysis.Chi-square test/t-test was used to compare and analyze the clinical data of the two groups.Results Among the 890 cases of pediatric brain injury,there were 519 males and 371 females.The median and quartiles of the age distribution for the two groups were as follows:MDT group 2.00(0.82,5.00)years and non-MDT group 1.00(1.00,4.00)years.Craniocerebral injury was the main type of brain injury in both groups,in addition,among children with craniocerebral injury and intracranial hemorrhage,the cure rate of MDT group was higher than that of non-MDT group,and the difference was statistically significant(P<0.05).Among the 405 children in MDT group,154(38.0%)underwent the surgery,while among the 485 children in non-MDT group,121(24.9%)underwent the surgery.The difference was statistically significant(P<0.05).23.2% of children in MDT group were in critical condition during the hospitalization,which was significantly lower than that in non-MDT group(30.5%),and the difference was statistically significant(P<0.05).The unhealed rate of MDT group(2.0%)was also significantly lower than that of non-MDT group(5.6%),the cure rate of MDT group(40.5%)was significantly higher than that of non-MDT group(34.4%),and there was a statistically significant difference(P<0.05).The expense of treatment,medicine and sanitary materials in MDT group were lower than those in non-MDT group,and the differences were statistically significant(P<0.05).The multivariate Logistic regression model analysis of the cure rate of children with brain injury showed that the MDT model could effectively improve the cure rate of children with brain injury(RR = 1.513,95% CI = 1.134-2.020).The results of multiple linear regression model analysis showed that there was no statistical difference in the effect of MDT on the actual hospitalization days of children(P>0.05).Conclusion Using MDT model to diagnose and treat children with brain injury is helpful to improve the cure rate,reduce the risk of children's disease aggravation,and achieve the significant therapeutic effects in children with brain injury.MDT model is worth popularizing and applying in children with brain injury.
3.Visualization analysis on research progress and trends of Ziziphi Spinosae Semen from 2000 to 2022
Tong QU ; Ning LI ; Hui REN ; Wenjing LU ; Xiaomin CUI ; Jing HU ; Zhiyong CHEN ; Hong ZHANG
China Pharmacist 2024;27(2):242-254
Objective To explore the research hotspots and development trends of Ziziphi Spinosae Semen in the past 20 years,and to provide reference for related research.Methods Literatures on Ziziphi Spinosae Semen were searched from January 1,2000 to December 31,2022 in CNKI,Web of Science Core Collection Database.VOS viewer software was used to visually analyze the citation frequency,research institutions and keyword hotspots of English literatures.CiteSpace software was used to visually analyze research institutions,authors,emergence keywords and keyword overlap time of Chinese literatures,and Microsoft Excel 2021 software was used to analyze of annual publication trends and publication volume of Chinese and English literatures,and download frequency of Chinese literatures.Results A total of 4 872 Chinese and 128 English literatures were included,with an overall upward trend in the number of annual publications.The research institutions with the highest number of publications in Chinese and English were Shandong University of Traditional Chinese Medicine and Tianjin University of Commerce,and the authors with the highest number of publications were DU Chenhui and XIE Junbo,respectively.The most frequent keywords in Chinese literatures were"Ziziphi Spinosae Semen","composed"and"application of compound therapy",and in English literatures were"performance","oxidative stress".Conclusion From 2000 to 2022,the research hotspots of Ziziphi Spinosae Semen mainly focused on the chemical composition,pharmacological effects and clinical application analysis,compatibility research,formulation and preparation.Quality control and evaluation of Ziziphi Spinosae Semen,and the research on the mechanism of preventing and treating insomnia with Ziziphi Spinosae Semen may become future research directions.
4.Changes of coagulation function and other indicators of the thawed FFP and FLP at 2-6℃
Jie PAN ; Xiangyun YAN ; Zhiyong LU ; Danhong WANG ; Qianqian CHEN ; Hongjie CHEN ; Yuting RUAN
Chinese Journal of Blood Transfusion 2024;37(9):1047-1051
【Objective】 To observe the changes of coagulation factor activity and protein content of the thawed fresh frozen plasma (FFP) and fresh liquid plasma (FLP) during storage at 2-6℃, and to provide reference for exploring the appropriate storage time of FFP at 2-6℃ after thawing. 【Methods】 The small-thaw group and the large-thaw group were respectively detected for the activity of coagulation factor FⅤ (FⅤ∶C) and FⅧ(FⅧ∶C), and the levels of fibrinogen (Fib), total protein (TP) and albumin (Alb) in TTP at 1, 2, 3, 4, 5, 6 and 7 days after thawing. And the FLP was detected for FⅤ∶C, FⅧ∶C, Fib, TP and Alb at 1, 2, 3, 4, 5, 6, 7 days and 1, 6, 11, 16, 21, 26 and 31 days after preparation, respectively. 【Results】 In FFP group, FⅧ∶C decreased gradually with the prolongation of storage time after melting (P<0.05), and decreased by 37.4% and 47.6% respectively in the two groups on the 7th day. There was no statistical difference in FⅤ∶C, Fib, TP and Alb (P>0.05). In FLP group, FⅧ∶C decreased gradually with the prolongation of storage time after melting (P<0.05). There was no statistical difference in FⅤ∶C in 7-day storage group (P>0.05), but it decreased gradually in 31-day storage group (P<0.05). There was no statistical difference in Fib, TP and Alb (P>0.05). 【Conclusion】 Although FⅧ∶C is decreased in thawed FFP stored at 2-6℃ for 7 days, it is still about 52.4%, which should be able to play a normal role in clinical practice.
5.Effectiveness and Safety of Tigecycline Combined with Cefoperazone-sulbactam Sodium in the Treatment of Multi-/extensively-drug Resistant Acinetobacter baumannii-associated Central Nervous System Infection:A Meta-analysis
Taojunjin LU ; Mingjuan ZHAO ; Wei WANG ; Zhiyong PAN ; Qin HU ; Yirong LI ; Zhiqiang LI
Herald of Medicine 2024;43(1):131-136
Objective To evaluate the efficacy and safety of tigecycline combined with cefoperazone-sulbactam sodium in the treatment of multi-/extensively-drug resistant Acinetobacter baumannii(MDRAB/XDRAB)associated central nervous system(CNS)infection,and to provide clinical evidence for antibiotic treatment of MDRAB/XDRAB-related intracranial disease.Methods The Wanfang Data Knowledge Service Platform,Chinese Biomedical Literature Database,VIP Chinese Science and Technology Journal Full-text Database,China National Knowledge Infrastructure(CNKI),Pubmed,Embase database,and Cochrane Library were searched to extract the literature of randomized controlled studies on tigecycline and cefoperazone sulbactam in the treatment of MDRAB/XDRAB CNS infection until September 1st,2022.The included studies were assessed for quality using the Cochrane Collaboration Risk of Bias assessment tool,and valid data were extracted and meta-analyzed using RevMan5.4 software.Results A total of 184 articles were screened and 4 Chinese RCTs were finally included,with a sample size of 267 cases.Meta-analysis showed that the overall efficacy of combination therapy for MDRAB/XDRAB CNS infection was better than monotherapy[OR = 4.30,95%CI =(1.93,9.58),P<0.01].Combination therapy had a better bacterial clearance[OR=4.20,95%CI=(2.08,8.48),P<0.01].And combination therapy resulted in a lower incidence of adverse effects[OR= 0.19,95%CI =(0.05,0.67),P<0.05].There was no apparent difference in cure rate between combination therapy and monotherapy(P>0.05).Conclusion Current evidence suggests that tigecycline combined with cefoperazone-sulbactam sodium may have better clinical efficacy and safety than monotherapy for MDRAB/XDRAB CNS infections.Limited by the number and quality of included studies,needs to be verified by more and higher-quality studies.
6.Prevalence of Lying Flat behaviors and its association with depressive symptoms among college students from three provinces and cities in China
Chinese Journal of School Health 2024;45(6):844-848
Objective:
To understand the prevalence of Lying Flat behaviors and its association with depressive symptoms among Chinese college students, so as to provide a scientific basis for promoting the physical and mental health development of adolescents.
Methods:
From July to October 2023, three universities were selected through convenient sampling from Jiangxi Province, Liaoning Province and Beijing City, respectively. Selfdesigned questionnaire links were distributed on campus to collect basic information and Lying Flat behaviors among college students, and the Patient Health Questionnaire-9 (PHQ-9) was utilized to screen for students with depressive symptoms. Finally, a total of 4 225 valid questionnaires were obtained. Chisquare was used to compare of report rates of Lying Flat behaviors across different demographic characteristics. Ordered Logistic regression analysis was used to explore the association between Lying Flat behaviors and depressive symptoms, with Z test used to assess variations in the strength of associations.
Results:
The reporting rates of academic, life, and social Lying Flat were 32.7%, 17.8% and 17.5%, respectively. And 6.7% of the participants were found of all three Lying Flat behaviors simultaneously.Among college students with three Lying Flat behaviors, the constituent ratios of no, mild, moderate and above depressive symptoms were 9.9%, 30.5% and 59.6%, respectively. Additionally, college students who had three Lying Flat behaviors were more likely to show mild, moderate and above depressive symptoms [OR(95%CI)=2.49(1.60-3.87), 7.69(5.01-11.79), P<0.01].
Conclusions
Academic Lying Flat behavior is most prevalent among college students. Academic, life and social Lying Flat behaviors are all significantly positively correlated with depressive symptoms. Attention should be paid to the Lying Flat behaviors and college students psychological health conditions to promote their physical and mental health development.
7.Long non-coding RNA AW112010 improves insulin resistance in adipocytes of aging mice through the miR-204/POU2F2 axis
Rui WANG ; Shuwen WANG ; Yifan ZHANG ; Yaqi HU ; Qi YUAN ; Yuan WEN ; Xiaoling CHEN ; Ting LU ; Ying ZHENG ; Zhiyong LIN ; Mengzhen XUE ; Yaqi WANG ; Fangqi XIA ; Leiqi ZHU ; Chengfu YUAN
Chinese Journal of Endocrinology and Metabolism 2024;40(1):44-52
Objective:To investigate whether long non-coding RNA(lncRNA) AW112010 can improve insulin resistance in aging adipocytes through the miR-204/POU2F2 signaling pathway.Methods:In vivo experiment: C57BL/6 mice were divided into young control group(4 months old) and aging model group(18 months old) based on body weight. The expression levels of AW112010, miR-204-5p, POU2F2, aging related indicators(p16, p21), and insulin signaling pathway genes [insulin receptor(INSR), insulin receptor substrate 1(IRS1), phosphatidylinositol kinase(PI3K), protein kinase B(AKT)] in epididymal adipose tissue were detected using real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting. In vitro experiment: Using adriamycin(ADR) to induce 3T3-L1 aging adipocyte model, β-gal staining was used to observe cellular senescence, and miR-204 inhibitor and miR-204 mimic small interfering RNA were successfully constructed and transfected into 3T3-L1 adipocytes. Results:RT-qPCR and Western blot results showed that compared with the young group, the expression of AW112010 in the adipose tissue of aging mice was increased, while the expression of miR-204-5p was decreased. The expressions of POU2F2, p16, and p21 in the adipose tissue of aging mice were increased, while the expressions of INSR, IRS1, PI3K, GLUT4 mRNA and protein were decreased. The β-gal stainging results showed that the number of 3T3-L1 senescent adipocytes induced by ADR was significantly increased, and the expression levels of AW112010, POU2F2, p16, and p21 in ADR-induced senescent adipocytes were increased compared with the control group, while the expression levels of miR-204-5p, INSR, IRS1, PI3K, GLUT4 were decreased, and remaining glucose in the culture medium was increased. Compared with control, overexpression of miR-204 resulted in decreased expressions of aging indicators p16, p21, and target gene POU2F2 while the expressions of INSR and GLUT4 were increased.Conclusion:Upregulation of lncRNA AW112010 in adipocytes of aging mice may induce insulin resistance by targeting miR-204-5p/POU2F2/IRS1.
8.Research progress on the mechanism of gut microbiota participating in diabetes nephropathy
Fei XU ; Jin CHEN ; Yuhan LU ; Zhiyong LI
Journal of Pharmaceutical Practice and Service 2024;42(5):181-184,197
With the increasing prevalence of diabetes,the prevention and treatment of diabetes nephropathy have become a worldwide problem.The molecular mechanism of the occurrence and development of diabetes nephropathy is still unclear,but many studies in recent years have shown that gut microbiota plays an important role in the progress on diabetes nephropathy.The research progress on the mechanism of gut microbiota participating in diabetes nephropathy was reviewed in this article.
9.Experts consensus on standard items of the cohort construction and quality control of temporomandibular joint diseases (2024)
Min HU ; Chi YANG ; Huawei LIU ; Haixia LU ; Chen YAO ; Qiufei XIE ; Yongjin CHEN ; Kaiyuan FU ; Bing FANG ; Songsong ZHU ; Qing ZHOU ; Zhiye CHEN ; Yaomin ZHU ; Qingbin ZHANG ; Ying YAN ; Xing LONG ; Zhiyong LI ; Yehua GAN ; Shibin YU ; Yuxing BAI ; Yi ZHANG ; Yanyi WANG ; Jie LEI ; Yong CHENG ; Changkui LIU ; Ye CAO ; Dongmei HE ; Ning WEN ; Shanyong ZHANG ; Minjie CHEN ; Guoliang JIAO ; Xinhua LIU ; Hua JIANG ; Yang HE ; Pei SHEN ; Haitao HUANG ; Yongfeng LI ; Jisi ZHENG ; Jing GUO ; Lisheng ZHAO ; Laiqing XU
Chinese Journal of Stomatology 2024;59(10):977-987
Temporomandibular joint (TMJ) diseases are common clinical conditions. The number of patients with TMJ diseases is large, and the etiology, epidemiology, disease spectrum, and treatment of the disease remain controversial and unknown. To understand and master the current situation of the occurrence, development and prevention of TMJ diseases, as well as to identify the patterns in etiology, incidence, drug sensitivity, and prognosis is crucial for alleviating patients′suffering.This will facilitate in-depth medical research, effective disease prevention measures, and the formulation of corresponding health policies. Cohort construction and research has an irreplaceable role in precise disease prevention and significant improvement in diagnosis and treatment levels. Large-scale cohort studies are needed to explore the relationship between potential risk factors and outcomes of TMJ diseases, and to observe disease prognoses through long-term follw-ups. The consensus aims to establish a standard conceptual frame work for a cohort study on patients with TMJ disease while providing ideas for cohort data standards to this condition. TMJ disease cohort data consists of both common data standards applicable to all specific disease cohorts as well as disease-specific data standards. Common data were available for each specific disease cohort. By integrating different cohort research resources, standard problems or study variables can be unified. Long-term follow-up can be performed using consistent definitions and criteria across different projects for better core data collection. It is hoped that this consensus will be facilitate the development cohort studies of TMJ diseases.
10.α2-macroglobulin alleviates glucocorticoid-induced avascular necrosis of the femoral head in mice by promoting proliferation, migration and angiogenesis of vascular endothelial cells
Qi ZHU ; Yunxiang LU ; You PENG ; Jiale HE ; Zeyu WEI ; Zhiyong LI ; Yuxian CHEN
Journal of Southern Medical University 2024;44(4):712-719
Objective To explore the mechanism underlying the protective effect of α2-macroglobulin (A2M) against glucocorticoid-induced femoral head necrosis. Methods In a human umbilical vein endothelial cell (HUVEC) model with injuries induced by gradient concentrations of dexamethasone (DEX;10-8-10-5 mol/L), the protective effects of A2M at 0.05 and 0.1 mg/mL were assessed by examining the changes in cell viability, migration, and capacity of angiogenesis using CCK-8 assay, Transwell and scratch healing assays and angiogenesis assay. The expressions of CD31 and VEGF-A proteins in the treated cells were detected using Western blotting. In BALB/c mouse models of avascular necrosis of the femoral head induced by intramuscular injections of methylprednisolone, the effects of intervention with A2M on femoral trabecular structure, histopathological characteristics, and CD31 expression were examined with Micro-CT, HE staining and immunohistochemical staining. Results In cultured HUVECs, DEX treatment significantly reduced cell viability, migration and angiogenic ability in a concentration- and time-dependent manner (P<0.05), and these changes were obviously reversed by treatment with A2M in positive correlation with A2M concentration (P<0.05). DEX significantly reduced the expression of CD31 and VEGF-A proteins in HUVECs, while treatment with A2M restored CD31 and VEGF-A expressions in the cells (P<0.05). The mouse models of femoral head necrosis showed obvious trabecular damages in the femoral head, where a large number of empty lacunae and hypertrophic fat cells could be seen and CD31 expression was significantly decreased (P<0.05). A2M treatment of the mouse models significantly improved trabecular damages, maintained normal bone tissue structures, and increased CD31 expression in the femoral head (P<0.05). Conclusion A2M promotes proliferation, migration, and angiogenesis of DEX-treated HUVECs and alleviates methylprednisolone-induced femoral head necrosis by improving microcirculation damages and maintaining microcirculation stability in the femoral head.


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