AIM:To investigate the mechanism of Piezo1 channel activation promoting the increase in intra-cellular Ca2+ concentration([Ca2+]i)of rat coronary artery smooth muscle cells(CASMCs).METHODS:The primary CASMCs of SD rats were cultured,and the expression and subcellular localization of Piezo1 in the cells were observed by immunofluorescence staining.The Piezo1 and stromal interaction molecule 1(STIM1)in CASMCs were knocked down by siRNA transfection,and the expression levels of the proteins were detected by Western blot.Utilizing laser confocal mi-croscopy,the change of[Ca2+]i in CASMCs was detected by Fluo-4 AM fluorescent probes.RESULTS:It was confirmed by immunofluorescence staining that the expression of Piezo1 existed in primary rat CASMCs.Immunofluorescence staining also showed that Piezo1 was co-located with sarco-/endoplasmic reticulum Ca2+-ATPase 2(SERCA2),mitochondrial outer membrane protein TOM20 and nuclear membrane protein lamin B1.Western blot results showed that the protein expres-sion levels of STIM1 and Piezo1 were significantly down-regulated by siRNA transfection(P<0.05).Compared with con-trol group,Yoda1,the agonist of Piezo1,could increase the extracellular Ca2+ influx of CASMCs(P<0.01).However,the Ca2+ influx mediated by Yoda1 was not affected by the inhibition of L-type calcium channels.Treatment with Yoda1 in-creased the intracellular Ca2+ release of CASMCs(P<0.01).However,inhibition of calcium channels on endoplasmic re-ticulum,ryanodine receptor and inositol 1,4,5-triphosphate receptor,did not affect intracellular Ca2+ release mediated by Yoda1.After the Ca2+ in endoplasmic reticulum was emptied using thapsigargin(TG),Yoda1 also mediated the Ca2+ re-lease of other organelles in CASMCs(P<0.01).After inhibition of L-type calcium channels,treatment with store-operated calcium channel(SOCC)inhibitor BTP2 or knockdown of STIM1 led to the decrease in extracellular Ca2+ influx of CASMCs mediated by Yoda1(P<0.01).Treatment with TG increased the release of Ca2+ from the endoplasmic reticulum of CASMCs after knockdown of Piezo1(P<0.05),but the extracellular Ca2+ influx mediated by TG was not affected.After inhibition of L-type calcium channels and SOCC,knockdown of Piezo1 led to the decreases in intracellular Ca2+ release and extracellular Ca2+ influx induced by Yoda1(P<0.01).CONCLUSION:The Piezo1 agonist orchestrates the influx of extracellular Ca2+ by activating Piezo1 channels on the cell membrane and inducing the indirect activation of SOCC.More-over,it facilitates the release of Ca2+ from organelles.Consequently,these pathways synergistically elevate the[Ca2+]i of rat CASMCs.

AIM:To investigate the influence of fat mass and obesity-associated(Fto)gene on the aberrant contraction of aortic smooth muscle in diabetes mellitus(DM)mice,and to explore the mechanism of Fto gene underlying the calcium regulation.METHODS:Smooth muscle-specific Fto gene knockout(FtoSMKO)mice were generated using Cre-loxP technology.The experiment involved 3 groups of mice:wild-type(WT)group,DM model group and FtoSMKO-DM group,with 15 mice in each group.In DM group and FtoSMKO-DM group,type 1 DM was induced by intraperitoneal injec-tion of streptozotocin.The mice in WT group were injected with equal volume of citric acid-sodium citrate buffer solution.The influences of different drugs on the contraction responses of aortic smooth muscle in mice were analyzed using a multi-myograph system.The expression level of FTO protein in the aortic tissues was detected by Western blot.RESULTS:(1)Compared with WT mice,the expression levels of FTO protein in the aortic tissues of DM mice were significantly in-creased(P＜0.01).(2)The expression level of FTO protein in smooth muscle was significantly decreased after knockout of Fto gene(P＜0.01).Compared with WT group,the mice in DM group exhibited a significant decrease in body weight and a marked increase in fasting blood glucose level(P＜0.05).There were no noticeable differences in body weight or fasting blood glucose level between FtoSMKO-DM group and DM group(P＞0.05).(3)The contraction responses of aortic smooth muscle in DM group were substantially increased by phenylephrine compared with WT group.Specifically,vaso-constriction responses mediated by non-L-type calcium channels and store-operated calcium channels(SOCC)were signifi-cantly enhanced in DM group.In addition,the responses mediated by inositol 1,4,5-trisphosphate receptors(IP3R),which facilitate calcium release from the sarcoplasmic reticulum,were significantly enhanced.However,the responses mediated by caffeine-activated ryanodine receptors(RyR),which also facilitate calcium release from the sarcoplasmic re-ticulum,were significantly inhibited(P＜0.05).(4)Compared with DM group,the phenylephrine-induced contraction re-sponses of aortic smooth muscle in FtoSMKO-DM group were greatly weakened(P＜0.05).In particular,the vasoconstriction responses mediated by non-L-type calcium channels and SOCC in FtoSMKO-DM group were greatly suppressed(P＜0.05),while those mediated by caffeine-activated RyR were dramatically boosted(P＜0.05).However,IP3R-mediated responses were not affected(P＞0.05).CONCLUSION:Smooth muscle-specific Fto gene knockout suppresses contractile hyperre-sponsiveness in the aortic smooth muscle of DM mice,which may be attributed to involvement of FTO protein in calcium regulation in the vascular smooth muscle.

OBJECTIVE To investigate the effects of Codonopsis pilosula polysaccharide （CPP） regulating the nuclear factor- erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） signaling pathway on gastric mucosal injury in rats with chronic atrophic gastritis （CAG）. METHODS Rats were randomly divided into control group， model group， CPP low-dose， medium-dose and high-dose groups （CPP 10， 20， 40 mg/kg）， and ML385 group （Nrf2 inhibitor ML385 30 mg/kg+CPP 40 mg/kg）， 10 rats per group. CAG rat model was established using N-methyl-N′- nitro-N-nitrosoguanidine combined with irregular diet， then they were given drugs for consecutive 6 weeks. HE staining was used to observe the pathological changes in gastric tissue morphology； the levels of serum gastrin （GAS）， motilin （MTL）， pepsin （PP）， as well as tumor necrosis factor-α （TNF-α）， interleukin-8 （IL-8） malondialdehyde （MDA） and superoxide dismutase （SOD） in gastric mucosal tissue were detected； TUNEL assay was used to observe gastric mucosal tissue cell apoptosis； immunohistochemical assay was adopted to observe the expressions of Nrf2 and recombinant Bcl2 associated X protein （Bax） in gastric mucosal tissue； Western blot was used to detect the expressions of Nrf2， HO-1， Bax and Bcl-2 proteins in gastric mucosal tissue. RESULTS Compared with the control group， the gastric mucosal tissue was damaged； the levels of GAS， MTL， PP and SOD， and the protein expressions of Nrf2， HO-1 and Bcl-2 were significantly reduced in model group （P＜0.05）， while the levels of MDA， TNF-α and IL-8， the cell apoptosis index， and the protein expression of Bax were significantly increased （P＜0.05）. Compared with model group， CPP low-dose， medium-dose and high- dose groups showed varying degrees of improvement in gastric mucosal histopathology； the levels of the quantitative indicators were significantly reversed （P＜0.05）. Nrf2 inhibitor ML385 significantly attenuated the improvement effect of high-dose CPP on the above indicators in CAG rats （P＜0.05）. CONCLUSIONS CPP can improve gastric mucosal injury in CAG rats， and inhibit oxidative stress， inflammatory response， and cell apoptosis. The mechanism of action may be related to the activation of Nrf2/HO-1 signaling pathway.

Objective To study the effects of different pellet feed hardness on the growth and reproduction,feed utilization rate,and environmental dust in laboratory mice.Methods One hundred of fifty 50 3-week-old SPF-grade C57BL/6JGpt and 150 ICR laboratory mice were randomly divided into three groups,with an equal number of males and females.They were fed diets with different hardness of 18.62 kg,23.15 kg,and 27.89 kg.Body weight,feed utilization rate,and dust levels in cages were recorded and calculated for mice aged 3-10 weeks.Forty-five 6-week-old male mice and ninety 4-week-old female mice from each strain were randomly divided into three groups and fed pellet feeds with three different hardness levels.After 2 weeks of adaptation to the same hardness feed,the mice were paired at a 1:2 male-to-female ratio and monitored for reproductive data for 3 months.Results At the age of 4 weeks,the body weight of male C57BL/6JGpt mice in 23.15 kg group was significantly higher than that in the 18.62 kg and 27.89 kg groups(P＜0.01),and the body weight of females in the 18.62 kg group was significantly higher than that in the 27.89 kg group(P＜0.05).There was no significant difference in body weight among ICR mice aged 3-10 weeks across different feed hardness groups(P＞0.05).For both strains,feed utilization rate for males was higher than that for females across different feed hardness groups at all weeks of age(P＜0.01).Compared to the 27.89 kg group,both the 18.62 kg and 23.15 kg groups showed a significant increase in the 50-mesh dust levels in cages for both strains aged 4-8 weeks(except for 7-week-old C57BL/6JGpt mice)(P＜0.05).For both C57BL/6JGpt and ICR mice,there was no significant difference in basic reproductive performance such as interval between the first litter and the monthly production index among the three feed hardness groups during the experimental period(P＞0.05).However,the monthly production index of C57BL/6JGpt mice first increased and then decreased with the increase of feed hardness,while that of ICR mice increased with increasing feed hardness,though these differences were not statistically significant(P＞0.05).Conclusion Different strains and genders had different tolerance to feed hardness.C57BL/6JGpt mice are more adapted to lower hardness feeds,while ICR mice are better suited to slightly higher hardness feeds.

Objective:To summarize the best evidence of exercise management of tumor-associated sarcopenia and provide reference for medical personnel to formulate individualized exercise intervention plans.Methods:BMJ Best Practice, UpToDate, Cochrane Library, Embase, PubMed, CINAHL, China Biology Medicine disc, China National Knowledge Infrastructure, Wanfang database, National Guideline Clearinghouse, Scottish Intercollegiate Guidelines Network, and other databases were searched by computer for literature on exercise intervention for patients with tumor-associated sarcopenia, including clinical decision making, guidelines, expert consensus, evidence summary and systematic reviews. The search period was from the establishment of the databases to April 23, 2023. Two researchers independently conducted literature quality evaluation, evidence extraction, and integration.Results:A total of 12 articles were included, including 4 guidelines, 3 expert consensus papers, and 5 systematic reviews. A total of 32 pieces of evidence were summarized from five aspects, including exercise evaluation, exercise timing, exercise plan, exercise management, and evaluation indicators.Conclusions:This study summarizes the best evidence for exercise intervention in patients with tumor-associated sarcopenia and suggestes that medical staff should apply the proof according to the clinical context and the patient's wishes.

Objective To investigate the influence of serum cystatin C (Cys-C) and hypersensitivity C-reactive protein (hs-CRP) levels on the 3-year survival of patients undergoing maintenance hemodialysis (MHD). Methods A total of 358 patients with chronic renal failure who underwent MHD at the Second Affiliated Hospital of Nantong University from April 2011 to October 2020 were selected as study subjects. General clinical data, pre-dialysis laboratory test indicators, and echocardiographic indicators 3 months after dialysis were recorded. The survival status of patients after 3 years of dialysis was followed up, and the general clinical data, pre-dialysis laboratory test indicators, and echocardiographic indicators 3 months after dialysis were compared between surviving and deceased patients. Univariate and multivariate Cox regression analyses were performed to screen influencing factors of 3-year survival in MHD patients. Results At the 3-year follow-up, of the 302 MHD patients' 203 survived, and 99 died. Statistically significant differences were observed in age, primary disease, diabetes status, congestive heart failure, statin use, antiplatelet drug use, diuretic use, dialysis mode, estimated glomerular filtration rate (eGFR) and gamma-glutamyl transferase (GGT), alkaline phosphatase (AKP), total bilirubin (TBIL), β₂-microglobulin (β₂-MG), creatinine (Cr), low-density lipoprotein cholesterol (LDL-C), hypersensitive C-reactive protein (hs-CRP), and serum phosphorus (P) levels between surviving patients and deaths(P < 0.05 or P < 0.01). Univariate Cox regression results showed that age of MHD patients, primary disease (diabetic nephropathy, hypertensive nephropathy, polycystic kidney disease, others), comorbidities(diabetes, congestive heart failure, other cardiovascular diseases), drug use (statins, antiplatelet drugs), dialysis method (hemodialysis, hemodialysis + perfusion), laboratory test indicators [GGT, AKP, TBIL, total bile acid (TBA), albumin (ALB), Cr, Cys-C, eGFR, apolipoprotein A (APO-A), hs-CRP]were all influential factors of 3-year survival rate of MHD patients. Multivariate Cox regression analysis revealed that age, primary disease, other cardiovascular diseases, dialysis mode, AKP, ALB, TBIL, Cys-C, and hs-CRP were significant influencing factors for the survival of patients with chronic renal failure (P < 0.05). Conclusion Serum Cys-C and hs-CRP levels before MHD in patients with chronic renal failure may be associated with their 3-year survival after dialysis treatment. High serum Cys-C may reduce the risk of poor clinical prognosis, while high serum hs-CRP may increase the risk of poor clinical prognosis.

ObjectiveThis study aimed to investigate the HIV genotypic subtypes and molecular transmission clusters among men who have sex with men （MSM） with newly reported HIV infections in Dehong Dai and Jingpo Autonomous Prefecture （Dehong Prefecture）， Yunnan Province， China， between 2010 and 2019. The study aimed to identify potential high-risk transmitters and provide reference data for screening， management， and intervention of infection sources. MethodsPlasma samples from newly reported HIV-positive MSM individuals in Dehong Prefecture between 2010 and 2019 were collected. The viral pol gene fragments were amplified， sequenced， and genotyped. Genetic distances （GD） between pairwise sequences were analyzed and calculated. MEGA 7.0 and Gephi were used for phylogenetic and molecular transmission network analysis. ResultsA total of 159 newly reported HIV infections among MSM were included in the study， with successful genotyping of 100 cases. Nine HIV-1 subtypes were identified， with the most prevalent being CRF01_AE subtype （52%）， followed by CRF07_BC subtype （31%）， CRF55_01B subtype （10%）， and others （7%）. Cluster analysis revealed a total network access rate of 67%， forming three transmission clusters. CRF01_AE subtype formed two transmission clusters with 38 and 3 infected individuals， while CRF07_BC subtypes formed one transmission cluster with 26 infected individuals. The transmission network within the CRF01_AE clusters exhibited a more complex relationship. Significant differences in educational level were observed between the two main transmission clusters. ConclusionThe predominant HIV subtypes among newly reported MSM cases in Dehong Prefecture between 2010 and 2019 were CRF01_AE and CRF07_BC. Significant cultural differences are observed between the main transmission clusters. Continued monitoring of genotypic subtypes and targeted interventions within transmission clusters are warranted.

Objective:To investigate the effect of volatile organic compounds (VOCs) exposure in the atmosphere on the risk of daily death from stroke in Guangzhou.Methods:Daily average concentrations of twelve atmospheric VOCs, meteorological factors, and daily deaths for stroke and its subtypes (including ischemic and hemorrhagic stroke) in Guangzhou from 2020 to 2021 were collected. The time-series Poisson generalized additive model was established to analyze the relationship between daily average concentrations of atmospheric VOCs and daily mortality from a stroke on different lag days. The season, gender, and age group further performed stratification analysis.Results:Toluene and n-pentane were associated with a higher mortality risk from stroke and its subtypes. For each interquartile range ( IQR) increment in toluene concentration at lag0- 1 days, the RRs for mortality from stroke and hemorrhagic stroke were 1.060 (95% CI: 1.036-1.085) and 1.071 (95% CI: 1.030-1.113), respectively. For each IQR increment in n-pentane concentration, the RR for mortality from ischemic stroke was 1.064 (95% CI: 1.030-1.099). The effect estimates of VOCs may be higher during the cold season and among women and people aged ≥75 years. For each IQR increment in toluene concentration, the RRs for mortality risk of stroke in the cold season and women were 1.099 (95% CI: 1.056-1.143) and 1.085 (95% CI: 1.050-1.120), respectively. For n-pentane, the RR for death risk of stroke in people aged ≥75 years old was 1.072 (95% CI: 1.036-1.109). Results of sensitivity analysis showed that the effect estimates fluctuated less when PM 2.5 and O 3 were separately introduced for the two-pollutant model, as well as changing the degrees of freedom for covariates. Conclusions:This study suggests that VOCs may be an independent risk factor for daily mortality from stroke. Moreover, Toluene presented the most significant health impact.

Severe asthma is "asthma which requires treatment with high dose inhaled corticosteroids (ICS) plus a second controller (and/or systemic corticosteroids) to prevent it from becoming 'uncontrolled’ or which remains 'uncontrolled’ despite this therapy." The state of control was defined by symptoms, exacerbations and the degree of airflow obstruction. Therefore, for the diagnosis of severe asthma, it is important to have evidence for a diagnosis of asthma with an assessment of its severity, followed by a review of comorbidities, risk factors, triggers and an assessment of whether treatment is commensurate with severity, whether the prescribed treatments have been adhered to and whether inhaled therapy has been properly administered. Phenotyping of severe asthma has been introduced with the definition of a severe eosinophilic asthma phenotype characterized by recurrent exacerbations despite being on high dose ICS and sometimes oral corticosteroids, with a high blood eosinophil count and a raised level of nitric oxide in exhaled breath. This phenotype has been associated with a Type-2 (T2) inflammatory profile with expression of interleukin (IL)-4, IL-5, and IL-13. Molecular phenotyping has also revealed non-T2 inflammatory phenotypes such as Type-1 or Type-17 driven phenotypes. Antibody treatments targeted at the T2 targets such as anti-IL5, anti-IL5Rα, and anti-IL4Rα antibodies are now available for treating severe eosinophilic asthma, in addition to anti-immunoglobulin E antibody for severe allergic asthma. No targeted treatments are currently available for non-T2 inflammatory phenotypes. Long-term azithromycin and bronchial thermoplasty may be considered. The future lies with molecular phenotyping of the airway inflammatory process to refine asthma endotypes for precision medicine.

ObjectiveTo determine the trend and influencing factors of HIV subtypes in newly reported HIV/AIDS cases in Dehong Dai and Jingpo Autonomous Prefecture （Dehong Prefecture） from 2017 to 2019. MethodsRNA extraction was conducted among newly reported HIV/AIDS cases in Dehong Prefecture from 2017 to 2019 whose plasma volume was more than 200 μL. The gag， env and pol genes were amplified by using RT-PCR and then sequenced to determine the subtypes. ResultsA total of 3 287 HIV-infected cases were newly reported in Dehong from 2017 to 2019. The HIV gag， env and pol genotypes were determined in 1 813 cases. The major subtypes were subtype C （28.4%，515/1 813）， recombination form BC （22.0%，398/1 813） and CRF_01AE （18.1%，329/1 813）. Furthermore， the proportion of subtype B， subtype C and CRF01_AE decreased over years， whereas 01/BC， CRF07_BC and CRF08_BC increased over years in both Chinese and Burmese patients （χ²=75.212，P<0.001）. There were significant differences in gender， age， marital status， ethnicity， educational level and transmission route among Chinese and Burmese HIV-infected cases with diverse HIV genotypes （all P<0.05）. ConclusionHIV subtypes in Dehong change over time， which demonstrates that the proportion of BC recombinant subtypes and unique recombinant subtypes increased significantly.