Objective:To investigate the prospective association between lifestyle and the risk for all-cause mortality in middle-aged and elderly residents in China.Methods:The data from China Health and Retirement Longitudinal Study were used. Baseline information about the lifestyle were collected through questionnaire survey and physical measurements, and the mortality data were obtained through surveys conducted at 2-3 year intervals. A total of 5 436 study participants were included. A comprehensive lifestyle including smoking, alcohol consumption, sleep, BMI and physical activity was constructed, and a multivariate-adjusted Cox proportional hazards regression model was used to estimate the association between lifestyle and the risk for all-cause mortality.Results:During the follow-up of average 8.2 years, 695 deaths were recorded. The comprehensive lifestyle score was linearly associated with the risk for all-cause mortality. Compared with the study participants with comprehensive lifestyle score of 0-1, those with score of 2-5 all had lower risk for all-cause mortality, with HRs of 0.78 (95% CI: 0.62-0.98), 0.56 (95% CI: 0.44-0.72), 0.36 (95% CI:0.27-0.48), and 0.33 (95% CI: 0.21-0.52), respectively. The results of Cox proportional hazards regression model analysis of single lifestyle showed that compared with those with unhealthy lifestyles, the HRs of all-cause mortality for study participants who never smoked, had moderate alcohol consumption, had appropriate night sleep, maintained healthy body weight and kept active physical activity were 0.70 (95% CI: 0.57-0.84), 0.76 (95% CI: 0.64-0.90), 0.79 (95% CI: 0.67-0.94), 0.73 (95% CI: 0.62-0.87), and 0.68 (95% CI: 0.58-0.80), respectively. Conclusions:Keeping healthy lifestyles can significantly reduce the risk for all-cause mortality in middle-aged and elderly residents China. The higher the healthy lifestyle level, the lower the risk for all-cause mortality.

Objective To investigate the effects and mechanisms of supplemented Buyang Huanwu decoction(BYHWD)on zinc home-ostasis and reproductive function in oligoasthenozoospermia(OAS)rats.Methods As many as 60 male SD rats were randomly divided into control,OAS,L-carnitine(0.323 g/kg),BYHWD low-dose(BYHWD-L,19.5 g/kg),BYHWD medium-dose(BYHWD-M,39 g/kg),and BYHWD high-dose(BYHWD-H,78 g/kg)groups,with 10 rats in each group.Except for the control group,all other groups were admini-stered adenine by gavage to establish an OAS model.After successful construction of the model,each group was treated by gavage with the corresponding drug doses or physiological saline once daily for 4 weeks.The general condition of the rats was recorded,testicular and epididymal coefficients were calculated,and sperm water volume and vitality were measured.Enzyme linked immunosorbent assay(ELISA)was used to measure the levels of serum testosterone(T),estradiol(E2),follicle-stimulating hormone(FSH),luteinizing hormone(LH),prolactin(PRL),as well as the levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor α(TNF-α)and zinc containing enzymes alcohol dehydrogenase(ADH),alkaline phosphatase(ALP),and lactate dehydrogenase(LDH)in testicular tissue.The zinc con-tent in testicular tissue were also measured.Pathological changes in testicular tissue were observed using hematoxylin eosin(HE)staining.Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear factor kappa-B(NF-κB)protein expression in testicu-lar tissue were identified using immunohistochemistry.Results Compared to the control group,the OAS group exhibited significantly lower testicular and epididymal coefficients,sperm motility,sperm count,serum T and E2 levels(P＜0.05)and significantly higher FSH,LH,and PRL levels(P＜0.05).Furthermore,the OAS group showed significantly lower testicular tissue zinc content and levels of zinc containing enzymes ADH,ALP,and LDH(P＜0.05)than the control group.Conversely,IL-1β,IL-6,and TNF-α levels,along with protein expressions of TLR4,MyD88,and NF-κB were significantly higher in the OAS group than in the control group(P＜0.05).Compared to the OAS group,each drug intervention group showed improvements in testicular and epididymal coefficients,sperm motility,sperm count,T and E2 levels,testicular zinc content and zinc containing enzymes(ADH,ALP,and LDH)levels.Furthermore,the intervention groups demonstrated reduced FSH,LH,PRL,IL-1β,IL-6,and TNF-α levels,and TLR4/MyD88/NF-κB signaling pathway protein expression(P＜0.05).Conclusion BYHWD improves the reproductive function of OAS rats,potentially by regulating zinc homeostasis and TLR4/MyD88/NF-κB signaling pathway,consequently reducing the inflammatory response.

Objective:To investigate the prospective association between lifestyle and the risk for all-cause mortality in middle-aged and elderly residents in China.Methods:The data from China Health and Retirement Longitudinal Study were used. Baseline information about the lifestyle were collected through questionnaire survey and physical measurements, and the mortality data were obtained through surveys conducted at 2-3 year intervals. A total of 5 436 study participants were included. A comprehensive lifestyle including smoking, alcohol consumption, sleep, BMI and physical activity was constructed, and a multivariate-adjusted Cox proportional hazards regression model was used to estimate the association between lifestyle and the risk for all-cause mortality.Results:During the follow-up of average 8.2 years, 695 deaths were recorded. The comprehensive lifestyle score was linearly associated with the risk for all-cause mortality. Compared with the study participants with comprehensive lifestyle score of 0-1, those with score of 2-5 all had lower risk for all-cause mortality, with HRs of 0.78 (95% CI: 0.62-0.98), 0.56 (95% CI: 0.44-0.72), 0.36 (95% CI:0.27-0.48), and 0.33 (95% CI: 0.21-0.52), respectively. The results of Cox proportional hazards regression model analysis of single lifestyle showed that compared with those with unhealthy lifestyles, the HRs of all-cause mortality for study participants who never smoked, had moderate alcohol consumption, had appropriate night sleep, maintained healthy body weight and kept active physical activity were 0.70 (95% CI: 0.57-0.84), 0.76 (95% CI: 0.64-0.90), 0.79 (95% CI: 0.67-0.94), 0.73 (95% CI: 0.62-0.87), and 0.68 (95% CI: 0.58-0.80), respectively. Conclusions:Keeping healthy lifestyles can significantly reduce the risk for all-cause mortality in middle-aged and elderly residents China. The higher the healthy lifestyle level, the lower the risk for all-cause mortality.

Objective To investigate the effects and mechanisms of supplemented Buyang Huanwu decoction(BYHWD)on zinc home-ostasis and reproductive function in oligoasthenozoospermia(OAS)rats.Methods As many as 60 male SD rats were randomly divided into control,OAS,L-carnitine(0.323 g/kg),BYHWD low-dose(BYHWD-L,19.5 g/kg),BYHWD medium-dose(BYHWD-M,39 g/kg),and BYHWD high-dose(BYHWD-H,78 g/kg)groups,with 10 rats in each group.Except for the control group,all other groups were admini-stered adenine by gavage to establish an OAS model.After successful construction of the model,each group was treated by gavage with the corresponding drug doses or physiological saline once daily for 4 weeks.The general condition of the rats was recorded,testicular and epididymal coefficients were calculated,and sperm water volume and vitality were measured.Enzyme linked immunosorbent assay(ELISA)was used to measure the levels of serum testosterone(T),estradiol(E2),follicle-stimulating hormone(FSH),luteinizing hormone(LH),prolactin(PRL),as well as the levels of interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor α(TNF-α)and zinc containing enzymes alcohol dehydrogenase(ADH),alkaline phosphatase(ALP),and lactate dehydrogenase(LDH)in testicular tissue.The zinc con-tent in testicular tissue were also measured.Pathological changes in testicular tissue were observed using hematoxylin eosin(HE)staining.Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),and nuclear factor kappa-B(NF-κB)protein expression in testicu-lar tissue were identified using immunohistochemistry.Results Compared to the control group,the OAS group exhibited significantly lower testicular and epididymal coefficients,sperm motility,sperm count,serum T and E2 levels(P＜0.05)and significantly higher FSH,LH,and PRL levels(P＜0.05).Furthermore,the OAS group showed significantly lower testicular tissue zinc content and levels of zinc containing enzymes ADH,ALP,and LDH(P＜0.05)than the control group.Conversely,IL-1β,IL-6,and TNF-α levels,along with protein expressions of TLR4,MyD88,and NF-κB were significantly higher in the OAS group than in the control group(P＜0.05).Compared to the OAS group,each drug intervention group showed improvements in testicular and epididymal coefficients,sperm motility,sperm count,T and E2 levels,testicular zinc content and zinc containing enzymes(ADH,ALP,and LDH)levels.Furthermore,the intervention groups demonstrated reduced FSH,LH,PRL,IL-1β,IL-6,and TNF-α levels,and TLR4/MyD88/NF-κB signaling pathway protein expression(P＜0.05).Conclusion BYHWD improves the reproductive function of OAS rats,potentially by regulating zinc homeostasis and TLR4/MyD88/NF-κB signaling pathway,consequently reducing the inflammatory response.

ObjectiveTo screen out the transcriptomes related to the intervention of Wuzi Yanzongwan on the spermatogenic function of semi-castrated male mice， and to explore its potential mechanism in the intervention of the progress of low spermatogenic function. MethodBalb/c mice were randomly divided into sham-operated group， model group， testosterone propionate group（0.2 mg·kg^-1·d^-1， intramuscular injection） and Wuzi Yanzongwan group（1.56 g·kg^-1·d^-1， intragastric administration） according to body weight， with 12 mice in each group. The right testicle and epididymis were extracted from the model group and the drug administration group to construct the semi-castrated model of low spermatogenic function， while the fur and the right scrotum of the sham-operated group were only cut and immediately sterilized and sutured. At the end of the intervention， hematoxylin-eosin（HE） staining was used to observe the histopathology of testis， enzyme-linked immunosorbent assay（ELISA） was used to detect the levels of serum testosterone（T）， luteinizing hormone（LH） and follicle stimulating hormone（FSH）. The sperm count and motility of epididymis were measured by automatic sperm detector of small animal. Transcriptomic microarray technology was used to detect the mRNA expression level of testicular tissue in each group， the transcriptome of genes related to the regulation of Wuzi Yanzongwan was screened， and three mRNAs were selected for Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） to verify the transcriptome data. Through the annotation analysis of Gene Ontology（GO） and the signaling pathway analysis of Kyoto Encyclopedia of Genes and Genomes（KEGG）， the related functions of drugs regulating transcriptome were analyzed. ResultCompared with the sham-operated group， the testicular tissue of mice in the model group showed spermatogenic injury， contraction and vacuolization of the seminiferous tubules， reduction of spermatogenic cells at all levels， widening of the interstitial space， obstruction of spermatogonial cell development and other morphological abnormalities， and serum T significantly decreased， LH significantly increased（P<0.01）， and FSH elevated but no statistically significant difference， the count and vitality of epididymal sperm significantly decreased（P<0.01）. There were 882 differentially expressed mRNAs in the testicular tissues， of which 565 were up-regulated and 317 were down-regulated. Cluster analysis showed that these differentially expressed mRNA could effectively distinguish between the sham-operated group and the model group. Compared with the model group， the damage to testicular tissue in the Wuzi Yanzongwan group was reduced， the structure of the seminiferous tubules was intact， vacuolization was reduced， and the number of spermatogenic cells at all levels was significantly increased and arranged tightly. The serum T significantly increased， LH significantly decreased（P<0.01）， and FSH decreased but the difference was not statistically significant. The count and vitality of sperm in the epididymis were significantly increased（P<0.01）. Moreover， Wuzi Yanzongwan could regulate 159 mRNA levels in the testes of semi-castrated mice， of which 32 were up-regulated and 127 were down-regulated， and the data of the transcriptome assay was verified to be reliable by Real-time PCR. GO and KEGG analysis showed that the transcriptome functions regulated by Wuzi Yanzongwan were involved in the whole cell cycle process of sperm development such as sex hormone production of interstitial cells in testis， renewal， differentiation， metabolism， apoptosis and signal transduction of spermatogenic cells， and were closely related to the biological behaviors of signaling pathways such as spermatogenic stem cell function， endoplasmic reticulum protein processing and metabolic program. ConclusionWuzi Yanzongwan can effectively improve the low spermatogenic function of semi-castrated male mice， and its mechanism may be related to the regulation of testicular transcriptional regulatory network， the synthesis of sex hormones in testicular interstitial cells， the function of spermatogenic stem cells， the whole cell cycle process of spermatogenesis， as well as the expression of endoplasmic reticulum protein processing and metabolic program related genes transcription.