ObjectiveTo explore the effects of transmembrane protein 16B （TMEM16B） on brain edema and blood-brain barrier after cerebral ischemia-reperfusion injury in rats. MethodsTMEM16B overexpression and knockdown was performed by adeno-associated virus （AAV）， and then adult male Sprague-Dawley rats were subjected to middle cerebral artery occlusion （MCAO）. Rats were randomly divided into Sham group， MCAO group， AAV no-load group， TMEM16B overexpression group and TMEM16B knockdown group. Modified neurological severity scores， adhesive removal test and cylinder test were used to evaluate neurologic function. The ultrastructure of ischemic brain tissue was observed by transmission electron microscope. Brain water content was reflected by dry wet weight ratio of brain tissue. The expressions of TMEM16B， aquaporin4 （AQP4）， Claudin5 and zonula occludens-1 （ZO-1） were investigated by immunofluorescence and Western blot. ResultsCompared with the AAV no-load group， the sensory and motor functions of rats in TMEM16B overexpression group were significantly impaired. Mitochondria were swollen； mitochondrial cristae and tight junctions disappeared. The brain water content was higher in overexpression group. The expression of TMEM16B and AQP4 increased while the expression of Claudin5 and ZO-1 decreased （all P<0.05）. Compared with the AAV no-load group， the rats in TMEM16B knockdown group showed some recovery in motor function. The mitochondrial cristae and structure were clear， and the basement membrane was partially blurred. The brain water content was lower in knockdown group. The protein levels of TMEM16B and AQP4 were lower while the levels of Claudin5 and ZO-1 were higher in TMEM16B knockdown group than in AAV no-load group （all P<0.05）. ConclusionAn increase in TMEM16B expression aggravates brain edema and blood-brain barrier damage in rats after cerebral ischemia-reperfusion injury， while a decrease in TMEM16B expression alleviates brain edema and protects the blood-brain barrier.

Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

Objective: To explore the correlation among picky eating levels in preschool children, parental self-efficacy and parenting stress. Methods: A convenience sampling method was employed to conduct an electronic questionnaire survey among 459 children aged 3-6 years and their parents from five kindergartens in Urumqi in November 2023. The survey included a general information questionnaire, the Children s Eating Behavior Questionnaire (CEBQ), the Parenting Sense of Competence Scale (PSOC), and the Parenting Stress Index-Short Form (PSI-SF). The Mann-Whitney U-test was used for twogroup comparisons, and the Kruskal-Wallis H-test was applied for multi-group comparisons. Spearman correlation analysis was conducted to examine the relationships between children s picky eating levels and parenting selfefficacy as well as parenting stress. Results: The picky eating score of preschool children was 10.00 (4.00), and the parenting self-efficacy score was 58.00 (12.00), both indicating a moderate level. The parenting stress score was 75.00 (16.00), reflecting a moderately low level. Spearman correlation analysis showed that children s picky eating levels were negatively correlated with the total score of parenting self-efficacy ( r =-0.28) and positively correlated with the total score of parenting stress( r =0.25)( P <0.01). Conclusions Picky eating levels of preschool children are closely associated with parenting self-efficacy and parenting stress. Picky eating behaviors in children can be reduced by implementing various effective measures to enhance parenting self-efficacy and alleviate parenting stress.

Following the core outcome set standards for development(COS-STAD), this study aims to construct core outcome set(COS) for clinical research on traditional Chinese medicine(TCM) treatment of simple obesity. Firstly, a comprehensive review was conducted on the randomized controlled trial(RCT) and systematic review(SR) about TCM treatment of simple obesity that were published in Chinese and English databases to collect reported outcomes. Additional outcomes were obtained through semi-structured interviews with patients and open-ended questionnaire surveys for clinicians. All the collected outcomes were then merged and organized as an initial outcome pool, and then a preliminary list of outcomes was formed after discussion by the working group. Subsequently, two rounds of Delphi surveys were conducted with clinicians, methodology experts, and patients to score the importance of outcomes in the list. Finally, a consensus meeting was held to establish the COS for clinical research on TCM treatment of simple obesity. A total of 221 RCTs and 12 SRs were included, and after integration of supplementary outcomes, an initial outcome pool of 141 outcomes were formed. Following discussions in the steering advisory group meeting, a preliminary list of 33 outcomes was finalized, encompassing 9 domains. Through two rounds of Delphi surveys and a consensus meeting, the final COS for clinical research on TCM treatment of simple obesity was determined to include 8 outcomes: TCM symptom scores, body mass index(BMI), waist-hip ratio, waist circumference, visceral fat index, body fat rate, quality of life, and safety, which were classified into 4 domains: TCM-related outcomes, anthropometric measurements, quality of life, and safety. This study has preliminarily established a COS for clinical research on TCM treatment of simple obesity. It helps reduce the heterogeneity in the selection and reporting of outcomes in similar clinical studies, thereby improving the comparability of research results and the feasibility of meta-analysis and providing higher-level evidence support for clinical practice.
Humans ; Obesity/therapy* ; Medicine, Chinese Traditional ; Randomized Controlled Trials as Topic ; Treatment Outcome ; Drugs, Chinese Herbal/therapeutic use*

This study aims to explore the mechanism of Buyang Huanwu Decoction(BHD) in promoting angiogenesis after oxygen-glucose deprivation/reoxygenation(OGD/R) of mouse brain microvascular endothelial cell line(brain-derived Endothelial cells.3, bEnd.3) based on the caveolin-1(Cav1)/Yes-associated protein 1(YAP1)/hypoxia-inducible factor-1α(HIF-1α) signaling pathway. Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used to analyze the blood components of BHD. The cell counting kit-8(CCK-8) method was used to detect the optimal intervention concentration of drug-containing serum of BHD after OGD/R injury of bEnd.3. The lentiviral transfection method was used to construct a Cav1 silent stable strain, and Western blot and polymerase chain reaction(PCR) methods were used to verify the silencing efficiency. The control bEnd.3 cells were divided into a normal group(sh-NC control group), an OGD/R model + blank serum group(sh-NC OGD/R group), and an OGD/R model + drug-containing serum group(sh-NC BHD group). Cav1 silent cells were divided into an OGD/R model + blank serum group(sh-Cav1 OGD/R group) and an OGD/R model + drug-containing serum group(sh-Cav1 BHD group). The cell survival rate was detected by the CCK-8 method. The cell migration ability was detected by a cell migration assay. The lumen formation ability was detected by an angiogenesis assay. The apoptosis rate was detected by flow cytometry, and the expression of YAP1/HIF-1α signaling pathway-related proteins in each group was detected by Western blot. Finally, co-immunoprecipitation was used to verify the interaction between YAP1 and HIF-1α. The results showed astragaloside Ⅳ, formononetin, ferulic acid, and albiflorin in BHD can all enter the blood. The drug-containing serum of BHD at a mass fraction of 10% may be the optimal intervention concentration for OGD/R-induced injury of bEnd.3 cells. Compared with the sh-NC control group, the sh-NC OGD/R group showed significantly decreased cell survival rate, cell migration rate, mesh number, node number, and lumen length, significantly increased cell apoptotic rate, significantly lowered phosphorylation level of YAP1 at S127 site, and significantly elevated nuclear displacement level of YAP1 and protein expression of HIF-1α, vascular endothelial growth factor(VEGF), and vascular endothelial growth factor receptor 2(VEGFR2). Compared with the same type of OGD/R group, the sh-NC BHD group and sh-Cav1 BHD group had significantly increased cell survival rate, cell migration rate, mesh number, node number, and lumen length, a significantly decreased cell apoptotic rate, a further decreased phosphorylation level of YAP1 at S127 site, and significantly increased nuclear displacement level of YAP1 and protein expression of HIF-1α, VEGF, and VEGFR2. Compared with the sh-NC OGD/R group, the sh-Cav1 OGD/R group exhibited significantly decreased cell survival rate, cell migration rate, mesh number, node number, and lumen length, a significantly increased cell apoptotic rate, a significantly increased phosphorylation level of YAP1 at S127 site, and significantly decreased nuclear displacement level of YAP1 and protein expression of HIF-1α, VEGF, and VEGFR2. Compared with the sh-NC BHD group, the sh-Cav1 BHD group showed significantly decreased cell survival rate, cell migration rate, mesh number, node number, and lumen length, a significantly increased cell apoptotic rate, a significantly increased phosphorylation level of YAP1 at the S127 site, and significantly decreased nuclear displacement level of YAP1 and protein expression of HIF-1α, VEGF, and VEGFR2. YAP1 protein was present in the protein complex precipitated by the HIF-1α antibody, and HIF-1α protein was also present in the protein complex precipitated by the YAP1 antibody. The results confirmed that the drug-containing serum of BHD can increase the activity of YAP1/HIF-1α pathway in bEnd.3 cells damaged by OGD/R through Cav1 and promote angiogenesis in vitro.
Drugs, Chinese Herbal/pharmacology* ; Animals ; Mice ; Signal Transduction/drug effects* ; Glucose/metabolism* ; Caveolin 1/genetics* ; Hypoxia-Inducible Factor 1, alpha Subunit/genetics* ; YAP-Signaling Proteins ; Oxygen/metabolism* ; Endothelial Cells/metabolism* ; Cell Line ; Adaptor Proteins, Signal Transducing/genetics* ; Neovascularization, Physiologic/drug effects* ; Cell Hypoxia/drug effects* ; Angiogenesis

Traditional four examinations of traditional Chinese medicine （TCM） are based on the symptoms and signs of patients， which are the advantages of TCM but also have shortcomings. Chronic kidney disease has the characteristics of insidiousness， long-term， deficiency and variability during its onset， which are difficult to be intervened in time based on only symptoms， therefore it is necessary to extend the application of the four examinations in the diagnosis and treatment process of chronic kidney disease. Based on the background of the continuous development of TCM syndrome differentiation techniques， this article proposed the extension and application strategies of the traditional four examinations in chronic kidney disease， including the incorporation of microscopic syndrome differentiation to identify the causes of kidney disease and prevent symptom deterioration； the utilization of accurate examination information enhanced by artificial intelligence for controlling development of existing disease； the integration of disease differentiation and syndrome differentiation to summarize clinical rules towards using constant to measure variation； and the establishment of a kidney disease database for the storage of four examinations information to prevent recurrence after recovery. The four above extension and application strategies can be used to achieve the long-term management and treatment effects of timely and early diagnosis， dynamic observation of the condition， accurate application of intervention， and strengthened prognosis assessment in the diagnosis and treatment of chronic kidney disease， and expand the advantages of TCM in the prevention and treatment of chronic kidney disease.

OBJECTIVE:High-intensity interval training(HIIT)and moderate-intensity continuous training(MICT)can improve the quality of life of patients with clinical chronic diseases,but their application effects and regulatory factors in adults with insufficient physical activity are still unclear.This study aimed to explore the application effects and regulatory factors of HIIT and MICT on the quality of life of adults with insufficient physical activity. METHODS:A systematic literature search was conducted in databases including Web of Science Core Collection,Medline(EBSCO Host),PubMed,and Cochrane Library.The search time limit was from the establishment of each database to September 2023.The types of included literature were randomized controlled trials,and the research subjects were physically inactive adults.RevMan 5.4 software and the GRADE evidence evaluation framework were used to assess the quality of the included literature.Main effects pooling of random effects models was performed using R Studio(version 4.2.0).Subgroup analyses,regression analyses,and sensitivity analyzes were used to explore the sources of study heterogeneity and moderators. RESULTS:(1)Thirty-two randomized controlled trials of moderate to high quality were included,involving 2 083 physically inactive adults(HIIT group n=474;MICT group n=708;control group n=901).(2)Compared with the non-training control group,HIIT[Hedges'g=0.61;95%confidence interval(CI):0.40-0.83;I2=45%]and MICT(Hedges'g=0.66;95%CI:0.25-1.08;I2=89%)significantly improved the quality of life.Direct comparison studies of HIIT and MICT found no significant differences in the quality of life(Hedges'g=-0.01;95%CI:-0.23-0.21;I2=0%).(3)Subgroup analysis showed that HIIT and MICT were more effective in improving the physical components of the quality of life(HIIT:Hedges'g=0.82 vs.0.75;MICT:Hedges'g=0.74 vs.0.55),while cycling had a better trend in improving overall quality of life(HIIT:Hedges'g=0.74 vs.0.36;MICT:Hedges'g=1.08 vs.0.52).(4)Additionally,regression analysis did not identify any significant moderators(P>0.05 for all factors).(5)None of the above meta-analyses found publication bias(Egger test P>0.05). CONCLUSION:(1)Moderate to high level evidence shows that both HIIT and MICT can improve the quality of life of adults with insufficient physical activity,and the intervention effects between the two are similar.Therefore,when choosing between these two options,it is necessary to comprehensively consider factors such as time economy,scheduling flexibility,and application feasibility to formulate a personalized exercise plan.(2)This study recommends that when applying HIIT,a low-volume protocol(for example,5 groups each time,1 minute each),3 times/week,and ride at 80%-95%of the maximum heart rate is used to achieve the theoretical best improvement effect.(3)Although MICT improves the quality of life,there is insufficient evidence that increasing exercise duration brings additional benefits.Therefore,this study recommends that when MICT is conducted,it should be carried out more than three times a week,with each training duration controlled between 25 and 60 minutes,and cycling at 50%-75%of the maximum heart rate,in order to achieve the theoretically expected best improvement effect.

Smart healthcare plays an important role in easing the strain on medical resources and improving the continuity of chronic disease management. This study analysed the iatrogenic risks from the intrinsic attributes and the external environment of smart healthcare, including doctor-patient conflict risk, technical operation risk, information leakage risk, humanistic absence risk, legal risk, regulatory risk and ethical risk. Based on the " structure process result" model, suggestions were proposed to optimize the construction of a smart healthcare platform for chronic diseases, improve the legal system and industry standards, strengthen talent cultivation and capacity building, establish an integrated regulatory system, and regularly evaluate the effectiveness of chronic disease management. These suggestions provided references for creating a healthy, orderly, and safe smart healthcare environment for chronic disease patients.

Objective:To investigate the association between hearing loss and the type of indoor fuel applications in Chinese middle-aged and elderly people through longitudinal cohort study.Methods:Data were obtained from the China Health and Retirement Longitudinal Study (CHARLS), including adults aged 45 years and older enrolled in 2011, with follow-up for cooking and heating analyses extending to 2018 and 2015, respectively. The study calculated the incidence of hearing loss based on an indoor cooking or heating fuel type and expressed in terms of per 100 person-years. The Cox proportional hazard model was used to assess the association between solid fuel use and hearing loss, and covariates such as gender, education, and economy were controlled. We also analyzed the impact of indoor fuel type and its switching on hearing loss.Results:A total of 6, 772 participants using household fuels for cooking (2011-2018) and 4, 618 for heating (2011-2015) were included. Those using solid fuels for cooking [(58.0±8.2) years] and heating [(58.1±8.5) years] were generally slightly older than that of those who used clean fuels. In the cooking analysis, the overall incidence of hearing loss was higher among solid fuel users compared to clean fuel users (Clean fuel: 2.6 cases per 100 person-years; solid fuel: 3.6 cases per 100 person-years; the difference between the two was statistically significant, P<0.05). However, no significant difference was observed in the heating analysis ( P>0.05). Further classification of fuel-type use revealed that the incidence of hearing loss was the highest among people who had been using solid fuels consistently. Compared to the clean fuel group, the fully adjusted hazard ratio (HR) was 1.5 (95% CI: 1.3-1.7) in the cooking analysis and 1.5 (95% CI: 1.1-2.0) in the heating analysis. Compared with using clean fuels, switching from clean fuels to solid fuels increased the risk of hearing loss both during cooking and heating processes. Conclusion:In the CHARLS database, individuals who use solid fuels for indoor cooking and heating are older than those who use clean fuels. Compared with clean fuel use, the use of solid fuels increases the risk of hearing loss in middle-aged and elderly people. Reducing the use of solid fuels, choosing clean fuels as substitutes for solid fuels, and avoiding the switch from clean fuels to solid fuels will help protect the hearing health of middle-aged and elderly individuals.

Researchers commonly use cyclization recombination enzyme/locus of X-over P1 (Cre/loxP) technology-based conditional gene knockouts of model mice to investigate the functional roles of genes of interest in Sertoli and Leydig cells within the testis. However, the shortcomings of these genetic tools include high costs, lengthy experimental periods, and limited accessibility for researchers. Therefore, exploring alternative gene silencing techniques is of great practical value. In this study, we employed adeno-associated virus (AAV) as a vector for gene silencing in Sertoli and Leydig cells. Our findings demonstrated that AAV serotypes 1, 8, and 9 exhibited high infection efficiency in both types of testis cells. Importantly, we discovered that all three AAV serotypes exhibited exquisite specificity in targeting Sertoli cells via tubular injection while demonstrating remarkable selectivity in targeting Leydig cells via interstitial injection. We achieved cell-specific knockouts of the steroidogenic acute regulatory ( Star ) and luteinizing hormone/human chorionic gonadotropin receptor (Lhcgr) genes in Leydig cells, but not in Sertoli cells, using AAV9-single guide RNA (sgRNA)-mediated gene editing in Rosa26-LSL-Cas9 mice. Knockdown of androgen receptor ( Ar ) gene expression in Sertoli cells of wild-type mice was achieved via tubular injection of AAV9-short hairpin RNA (shRNA)-mediated targeting. Our findings offer technical approaches for investigating gene function in Sertoli and Leydig cells through AAV9-mediated gene silencing.
Animals ; Male ; Leydig Cells/metabolism* ; Mice ; Dependovirus/genetics* ; Sertoli Cells/metabolism* ; Gene Silencing ; Genetic Vectors ; Testis/cytology*