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.Improvement effects of poria acid on insulin resistance in rats with polycystic ovary syndrome and its mechanism
Hong TANG ; Linxia LI ; Yu HUA ; Xiaomei JIANG ; Shiwen ZHANG ; Jinrong FU
China Pharmacy 2024;35(1):57-62
OBJECTIVE To study the improvement effects of poria acid on insulin resistance in rats with polycystic ovary syndrome (PCOS) and its mechanism. METHODS One hundred and twenty-six female rats were randomly separated into blank group, PCOS group, poria acid low-dose group (8.33 mg/kg), pachymic acid high-dose group (33.32 mg/kg), ethinylestradiol cyproterone group (positive control group, 0.34 mg/kg), recombinant rat high mobility group protein B1 protein (rHMGB1) group (8 μg/kg), and poria acid high dose+rHMGB1 group (33.32 mg/kg poria acid+8 μg/kg rHMGB1), with 18 rats in each group. Except for the blank group, the rats in all other groups were given Letrozole suspension intragastrically to construct the PCOS model. After successful modeling, administration was performed once a day for 4 weeks. After medication, the fasting blood glucose and fasting insulin levels, and insulin resistance index (HOMA-IR) were measured in rats; the levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone (T) in rat serum, and the levels of interleukin-1β (IL-1β) and tumor necrosis factor- α (TNF- α) in ovarian tissue were detected; ovarian coefficients of rats were calculated; the pathological changes of ovarian tissue were observed; the expressions of HMGB1, receptor for advanced glycosylation elaine_ tanghong@sina.com end product (RAGE) and phosphorylated nuclear factor κB p65 (p-NF-κB p65) proteins were determined in ovarian tissue of rats. RESULTS Compared with the blank group, the pathological injury of ovarian tissue of rats in the PCOS group was serious, the levels of fasting blood glucose and fasting insulin, HOMA-IR and ovarian coefficient were increased, the levels of serum LH and T were increased, while the levels of FSH were decreased; the levels of IL-1β and TNF-α, the expressions of HMGB1, RAGE and p-NF-κB p65 protein in ovarian tissue were increased, with statistical significance (P<0.05). Compared with the PCOS group, pathological damage of ovarian tissue was reduced in poria acid low-dose and high-dose groups and ethinylestradiol cyproterone group, and fasting blood glucose, fasting insulin levels, HOMA-IR and ovarian coefficient were decreased; serum LH and T levels were decreased, while FSH levels were increased; the levels of IL-1β and TNF-α and the expressions of HMGB1, RAGE and p-NF-κB p65 protein in ovarian tissue were decreased, with statistical significance (P<0.05). The trend of corresponding indexes in rHMGB1 group was opposite to the above (P<0.05). Compared with poria acid high-dose group, the changes of the above indexes were reversed significantly in poria acid high-dose+rHMGB1 group (P<0.05). CONCLUSIONS Poria acid may improve insulin resistance and inhibit inflammatory reaction in PCOS rats by inhibiting HMGB1/ RAGE pathway.
3.Status quo of preterm infants′ feeding difficulties at weaning and self-feeding transition stage and its influencing factors
Die CHEN ; Wentao PENG ; Mengyan TANG ; Xiaomei LIU
Chinese Journal of Child Health Care 2024;32(1):21-25
【Objective】 To investigate the prevalence and influencing factors of feeding difficulties in preterm infants at weaning and self-feeding transition stage, so as to provide a scientific basis for the management of feeding preterm infants. 【Methods】 Preterm infants at corrected age of 6 - 24 months were recruited from the Department of Child Health of five maternal and child health hospital of Chengdu from April to May 2021, and were surveyed by using the Chinese Version of the Montreal Children Hospital Feeding Scale (MCF-FS) and the self-designed questionnaire on the influencing factors of feeding difficulties. Then the status quo of feeding difficulties and its influencing factors were analyzed. 【Results】 The prevalence rate of feeding difficulties in 231 preterm infants was 32%. Among them, the prevalence rate of mild, moderate and severe feeding difficulties was 15.2%, 7.8% and 9.1%, respectively. Binary Logistic stepwise regression analysis indicated that food allergy (OR=4.253, 95%CI: 1.430 - 12.649), anxious mood of caregivers (OR=6.064, 95%CI: 2.998 - 12.268), tease or chase during eating(OR=2.873, 95%CI: 1.382 - 5.970), recreational activities at eating (OR=2.328, 95%CI: 1.115 - 4.860), and forced feeding (OR=2.772, 95%CI: 1.239 - 6.198) were positively associated with feeding difficulty of preterm infants(P<0.05). 【Conclusion】 Feeding difficulties in the weaning and self-feeding transition period of preterm infants are prevalent, so the guidance should focus on premature infants with food allergy, anxious caregivers and improper feeding behaviors, and appropriate interventions should be taken to promote scientific feeding.
4.Research progress in food neophobia in children
Xiao FU ; Mengyan TANG ; Xiaomei LIU
Chinese Journal of Child Health Care 2024;32(1):68-72
Food neophobia is one of the distinctive feeding disorders in children. It affects children′s physical growth and neurological development, resulting in negative impacts on their eating behaviors and habits. This review elucidates the concept and origin of food neophobia, and summarizes the prevalence, associated factors and effective interventions according to previous literature, in order to provide suggestions and guidance for the prevention and early intervention of children with food neophobia.
5.Comparison of the predictive value of multiparametric MRI and prostate-specific membrane antigen PET/CT for pelvic lymph node metastasis in prostate cancer
Wei TANG ; Yi CAI ; Yongxiang TANG ; Xiaoping YI ; Xiaomei GAO ; Lin QI
Chinese Journal of Surgery 2024;62(11):1008-1015
Objective:To compare the diagnostic performance of multiparametric magnetic resonance imaging (mpMRI) and prostate-specific membrane antigen (PSMA) PET/CT in detecting pelvic lymph node metastasis in prostate cancer.Methods:This is a retrospective case series study. A retrospective analysis was conducted on the data of 115 prostate cancer patients who underwent both mpMRI and PSMA PET/CT before undergoing radical prostatectomy and extended pelvic lymph node dissection at the Department of Urology, Xiangya Hospital, Central South University, between March 2020 and September 2023. The age ( M(IQR)) was 67(10) years (range: 45 to 84 years), and the body mass index was 24(4) kg/m 2 (range: 18 to 30 kg/m 2). Pathological and imaging data were obtained from the patients. Lymph node pathology results were used as the gold standard to evaluate the diagnostic performance of mpMRI and PSMA PET/CT for detecting pelvic lymph node metastasis in PCa through diagnostic evaluation tests. Comparisons between groups were performed using independent samples t-test, Mann-Whitney U test, or χ2 test. Results:The positive rate for detecting pelvic lymph node metastasis was 18.3% (21/115) with mpMRI and 25.2% (29/115) with PSMA PET/CT. The pathological positive rate for lymph nodes was 28.7% (33/115). In patient-based analysis, the diagnostic sensitivity of PSMA PET/CT was significantly higher than that of mpMRI (63.6% vs. 30.3%, χ2=7.36, P=0.007). In lesion-based analysis, both the sensitivity and positive predictive value of PSMA PET/CT were significantly higher than those of mpMRI (sensitivity: 68.0% vs. 21.6%, χ2=42.20, P<0.01; positive predictive value: 50.0% vs. 23.1%, χ2=7.54, P=0.006). Conclusions:PSMA PET/CT and mpMRI both demonstrates good specificity in predicting pelvic lymph node metastasis in prostate cancer. However, PSMA PET/CT is significantly superior to mpMRI in terms of sensitivity and the detection rate of pathologically positive lymph nodes.
6.Clinical characteristics and genetic analysis of four children with Rotor syndrome
Yanfang TAN ; Wenxian OUYANG ; Tao JIANG ; Lian TANG ; Hui ZHANG ; Ying YU ; Xiaomei QIN ; Shuangjie LI
Chinese Journal of Medical Genetics 2024;41(6):715-719
Objective:To explore the characteristics of SLCO1B1/ SLCO1B3 gene variants among children with Rotor syndrome (RS). Methods:Four children who were admitted to the Department of Hepatology of Hunan Children′s Hospital between January 2019 and January 2022 were selected as the study subjects. Trio-whole exome sequencing was carried out for the four families, and gel electrophoresis was used to verify an insertional variant of long-interspersed element-1 (LINE-1).Results:Genetic testing has identified three variants of the SLCO1B1 gene, including c. 1738C>T (p.R580*), c. 757C>T (p.R253*) and c. 1622A>C (p.Q541P), and two variants of the SLCO1B3 gene, including c. 481+ 22insLINE-1 and c. 1747+ 1G>A among the children. Three of them were found to harbor homozygous variants of the SLCO1B1/ SLCO1B3 genes, and one has harbored compound heterozygous variants. Sanger sequencing confirmed the existence of all variants, and gel electrophoresis has confirmed the existence of the LINE-1 insertional variant of about 6 kb within intron 6 of the SLCO1B3 gene in all children. Conclusion:The pathogenesis of the RS among the four children may be attributed to the variants of the SLCO1B1/ SLCO1B3 genes. The LINE-1 insertion variant of the SLCO1B3 gene may be common among Chinese RS patients.
7.Research progress on the etiology and diagnosis and treatment of anti NMDAR encephalitis
Zhenhong XU ; Xin LING ; Xiaomei DUAN ; Beisha TANG
Journal of Chinese Physician 2024;26(7):990-994
Anti N-methyl-D-aspartate receptor (NMDAR) encephalitis is a rare autoimmune encephalitis, and its diverse etiology and clinical manifestations have attracted attention. However, the etiology and pathogenesis of anti NMDAR encephalitis are not yet fully understood, and its diagnosis and treatment still have certain limitations. This review mainly explores the environmental genetic factors that drive the occurrence of diseases and the biological indicators that are beneficial for diagnosis. It also elaborates on the current immune regulation and emerging treatment pathways, points out potential targeted interventions, and looks forward to the challenges and future development directions they face.
8.Common Malignant Tumors in the Reproductive System of Chinese Women:Disease Burden During 1990-2019 and Prediction of Future Trend
Nan ZHENG ; Jun TANG ; Xiaomei REN
Acta Academiae Medicinae Sinicae 2024;46(1):25-32
Objective To analyze the trends of disease burden of cervical cancer,uterine cancer,and ovarian cancer among Chinese women from 1990 to 2019,and to provide a basis for formulating precise preven-tion and control measures in China.Methods The global disease burden data in 2019 were used to describe the changes in indicators such as incidence,mortality,years of life lost due to premature mortality(YLL),years lived with disability(YLD),and disability-adjusted life year(DALY)of cervical,uterine,and ovarian canc-ers in China from 1990 to 2019.Furthermore,the Bayesian age-period-cohort model was adopted to predict the incidence and mortality of the cancers from 2020 to 2030.Results From 1990 to 2019,the incidence rates and mortality of cervical,uterine,and ovarian cancers in Chinese women showed an upward trend,and the age-standardized incidence rate of ovarian cancer increased the most(0.78% ).In 2019,the incidence of cervical cancer and uterine cancer concentrated in the women of 55-59 years old,and ovarian cancer mainly occurred in the women of 70-74 years old.The DALY,YLL,and YLD of cervical,uterine,and ovarian cancers all pres-ented varying degrees of growth at all ages.The Bayesian age-period-cohort model predicted that from 2020 to 2030,the incidence and mortality of cervical cancer in China showed a decreasing trend,while those of uterine cancer and ovarian cancer showed an increasing trend.There was no significant change in the age with high inci-dence of the three cancers.Conclusions From 1990 to 2019,the overall disease burden of cervical,uterine,and ovarian cancers in China increased,while the disease burden of cervical cancer decreased after 2020.It is recom-mended that the efforts should be doubled for the prevention and control of cervical,uterine,and ovarian cancers.
9.Status and associated factors of the knowledge and health education needs about sexual abuse of middle school students in rural areas
TANG Zhengyan, ZHANG Yongai, LI Xiaomei
Chinese Journal of School Health 2024;45(7):969-973
Objective:
To explore the status and associated factors of the knowledge and health education needs about child sexual abuse of middle school students by sex, so as to provide a basis for targeted safety education of preventing child sexual abuse.
Methods:
From June to July 2023, 1 256 junior high school students from 2 middle schools in southern area of Shaanxi Province were selected by a convenience sampling method. A selfdesigned general information questionnaire, Childrens Sexual Assault Cognition Questionnaire, and Health Education Needs Questionnaire were used for questionnaire survey. Group comparisons were conducted using ttests, analysis of variance, and Chisquare tests. Multiple linear regression analysis was used to explore the factors related to cognitive scores of sexual assault, and binary Logistic regression analysis was used to explore the factors related to the willingness of middle school students to receive sexual assault safety education.
Results:
The correct response rate of the participants for knowledge of sexual assault cognition (CSA) was (80.97±12.09)%. For each item, the correct response rate ranged 46.7%-97.2%. The correct rate (78.98±12.23)% of male students knowledge of CSA was lower than female students (83.17±11.55)% statistically (t=-6.23, P<0.01). The factors influencing the participants knowledge of CSA included gender (β=0.16), mothers occupation (β=0.07) and experience of CSA safety education (β=0.10) (P<0.05). Most students (92.7%) were willing to receive CSA education. Girls (OR=1.72, 95%CI=1.06-2.77) and students who previously received CSA education (OR=6.98, 95%CI=4.44-10.96) had more willingness to receive CSA education (P<0.05). A total of 71.8% of middle school students preferred their parents as educators, while 43.4% of students chose onsite instruction as the preferred method of CSA education.
Conclusions
Middle school students have a strong willingness to accept CSA safety education. Knowledge of CSA and willingness to receive CSA safety education are related to gender and previous educational experience.
10.Analysis of the molecular epidemiological characteristics of carbapenem-resistant Klebsiella pneumoniae in a hospital in Hunan Province
Xingwang NING ; Yongxue TANG ; Siyu WANG ; Xiaomei WANG ; Huibin ZHU ; Xiaobing XIE ; Qingyu LIU
Chinese Journal of Preventive Medicine 2024;58(7):1041-1047
To examine the molecular epidemiology of carbapenem-resistant Klebsiella pneumoniae (CRKP) and investigate the horizontal transmission of blaKPC and blaNDM genes for the prevention and treatment of CRKP. A total of 49 clinically isolated CRKP strains were retrospectively analyzed from January to December 2022 at The First Hospital of Hunan University of Chinese Medicine. Phenotypic screening was performed using modified carbapenem inactivation assay (mCIM) and EDTA-carbapenem inactivation assay (eCIM). Polymerase chain reaction (PCR) was utilized to identify carbapenem resistance genes, β-lactamase resistance genes, and virulence genes, while multi-locus sequence analysis (MLST) was employed to assess the homology of CRKP strains. Conjugation experiments were conducted to infer the horizontal transmission mechanism of blaKPC and blaNDM genes. The results showed that the study included 49 CRKP strains, with 44 carrying blaKPC and 8 carrying blaNDM, Three strains were identified as blaKPC+ blaNDM-CRKP. In this study, 28 out of 49 CRKP strains (57.2%) were found to carry virulence genes. Additionally, one CRKP strain tested positive in the string test and was found to carry both Aerobactin and rmpA virulence genes. MLST results revealed a total of 5 ST types, with ST11 being predominant (41/49, 83.7%). Successful conjugation was observed in all 3 blaKPC-CRKP strains, while only 1 out of 3 blaNDM-CRKP strains showed successful conjugation. The transconjugant exhibited significantly reduced susceptibility to imipenem and cephalosporin antibiotics. In conclusion, the resistance mechanism of CRKP in this study is primarily attributed to the production of KPC enzymes, along with the presence of multiple β-lactamase resistance genes. Additionally, there is a local prevalence of hv-CRKP and blaKPC+ blaNDM-CRKP. blaKPC and blaNDM can be horizontally transmitted through plasmids, with varying efficiency among different strains.


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