BACKGROUND:Forskolin,a diterpenoid natural compound extracted from Coleus forskohlii,has a crucial regulatory role in skeletal muscle repair.However,the regulatory role of forskolin on myogenic differentiation of C2C12 skeletal muscle cells has not been fully explored.OBJECTIVE:To explore the effects of forskolin on the differentiation of C2C12 myoblast cell line and probe into the underlying molecular mechanisms.METHODS:C2C12 cells were treated with 0,0.1,0.25,0.5,1,5,10 and 20 μmol/L forskolin during growth,and cell proliferation was detected by cell counting kit-8 and qRT-PCR.C2C12 cells were treated with 0,0.25,0.5 and 1 μmol/L forskolin during the induction of myogenic differentiation.Immunofluorescence staining and qRT-PCR were used to detect C2C12 cells differentiation.Western blot was used to detect the expression level of myogenic differentiation-related signaling pathway proteins.RESULTS AND CONCLUSION:(1)The viability of C2C12 cells was decreased and cell proliferation was inhibited after treatment with high concentrations(＞1 μmol/L)of forskolin.(2)The qRT-PCR results showed that forskolin up-regulated the expression of Myh2,Myh4,Myomaker,but down-regulated the expression of Myh7 compared with the 0 μmol/L group,when C2C12 cells were differentiated for 4 days.Immunofluorescence staining results showed that the fusion index and myotube diameter of C2C12 cells were increased after forskolin treatment,and the number of myotubes was also increased.(3)Western blot results showed that the phosphorylated extracellular signal-regulated kinase 1/2 expression was inhibited;however,the phosphorylated protein kinase B was promoted after treatment with forskolin.The protein expression level of the myogenic differentiation transcription factor Myogenin was significantly up-regulated after treatment with forskolin.The above results demonstrate that forskolin may promote myogenic differentiation of C2C12 skeletal muscle cells through the extracellular signal-regulated kinase 1/2 and protein kinase B signaling pathway.

BACKGROUND:Forskolin,a diterpenoid natural compound extracted from Coleus forskohlii,has a crucial regulatory role in skeletal muscle repair.However,the regulatory role of forskolin on myogenic differentiation of C2C12 skeletal muscle cells has not been fully explored.OBJECTIVE:To explore the effects of forskolin on the differentiation of C2C12 myoblast cell line and probe into the underlying molecular mechanisms.METHODS:C2C12 cells were treated with 0,0.1,0.25,0.5,1,5,10 and 20 μmol/L forskolin during growth,and cell proliferation was detected by cell counting kit-8 and qRT-PCR.C2C12 cells were treated with 0,0.25,0.5 and 1 μmol/L forskolin during the induction of myogenic differentiation.Immunofluorescence staining and qRT-PCR were used to detect C2C12 cells differentiation.Western blot was used to detect the expression level of myogenic differentiation-related signaling pathway proteins.RESULTS AND CONCLUSION:(1)The viability of C2C12 cells was decreased and cell proliferation was inhibited after treatment with high concentrations(＞1 μmol/L)of forskolin.(2)The qRT-PCR results showed that forskolin up-regulated the expression of Myh2,Myh4,Myomaker,but down-regulated the expression of Myh7 compared with the 0 μmol/L group,when C2C12 cells were differentiated for 4 days.Immunofluorescence staining results showed that the fusion index and myotube diameter of C2C12 cells were increased after forskolin treatment,and the number of myotubes was also increased.(3)Western blot results showed that the phosphorylated extracellular signal-regulated kinase 1/2 expression was inhibited;however,the phosphorylated protein kinase B was promoted after treatment with forskolin.The protein expression level of the myogenic differentiation transcription factor Myogenin was significantly up-regulated after treatment with forskolin.The above results demonstrate that forskolin may promote myogenic differentiation of C2C12 skeletal muscle cells through the extracellular signal-regulated kinase 1/2 and protein kinase B signaling pathway.

Objective This study examined changes in serum growth hormone(GH)and insulin-like growth factor 1(IGF-1)levels,as well as insulin resistance,in patients with type 2 diabetes mellitus(T2DM)across different stages of diabetic kidney disease(DKD).It explored the relationship between these factors and the occurrence and progression of DKD,with the aim of identifying new targets for the prevention and treatment of DKD.Methods In this cross-sectional study,T2DM patients aged 18-80 years were selected from the Department of Endocrinology,The Second Affiliated Hospital of Xi'an Jiaotong University,from November 30,2019 to March 30,2021.Their serum GH and IGF-1 levels were tested.Results The serum GH level increased gradually in the groups,and the difference was statistically significant(P＜0.05).The level of serum IGF-1 decreased first and then increased among the groups,but with no significant difference(P＞0.05).HOMA-IR showed no significant correlation among the groups(P＞0.05),but HOMA-β gradually decreased among the groups,with statistically significant difference(P＜0.05).Multivariate Logistic regression analysis showed that the increase of serum GH level was closely related to the progression of T2DM to early DKD,and that the increase of serum IGF-1 level might be related to the deterioration of renal function in patients with T2DM at DKD stage.HOMA-IR and HOMA-β were not found to have correlation with the occurrence or development of DKD in T2DM patients.Conclusion In patients with T2DM,elevated serum GH level is closely associated with the occurrence and progression of early DKD.The increase of serum GH level may promote the occurrence of early DKD in T2DM,and the increase of serum IGF-1 level may be related to the deterioration of renal function in clinical DKD stage.In this study,we did not find correlation of HOMA-IR and HOMA-β with the occurrence or development of DKD in T2DM patients.

Objectives:To investigate the clinical and cardiopulmonary functional characteristics of cardiopulmonary-phenotype idiopathic pulmonary arterial hypertension(IPAH)patients in comparison with classical IPAH patients and pulmonary hypertension patients associated with chronic lung disease(CLD-PH).Methods:In this retrospective study,data were collected from 30 patients with classical IPAH,20 cardiopulmonary phenotype IPAH patients,and 20 patients with CLD-PH,who were hospitalized in the Department of Respiratory and Critical Care Medicine at Beijing Chaoyang Hospital from November 2017 to February 2025.Pulmonary hypertension was diagnosed via right heart catheterization and all patients underwent pulmonary function tests,chest computed tomography(CT),echocardiography,and patients were followed up to 5 years.Results:Compared to classical IPAH patients,both cardiopulmonary phenotype IPAH patients and CLD-PH patients exhibited later onset age,higher proportions of World Health Organization(WHO)functional class Ⅲ-Ⅳ,males,and smokers(all P<0.05).The cardiopulmonary-phenotype IPAH patients also had higher rates of coronary artery disease and diabetes compared to classical IPAH patients(all P<0.05).Physiologically,the cardiopulmonary-phenotype IPAH patients showed reduced diffusing capacity for carbon monoxide(DLCO)and partial pressure of oxygen(PaO2),along with higher rates of emphysema and pulmonary fibrosis compared to classical IPAH(all P<0.05).In contrast,CLD-PH patients had lower mean pulmonary arterial pressure(mPAP),pulmonary vascular resistance(PVR),and pulmonary artery wedge pressure(PAWP),as well as reduced forced expiratory volume in the first second(FEV1%predicted),FEV1/forced vital capacity(FVC)ratio,and DLCO.However,CLD-PH patients demonstrated higher tricuspid annular plane systolic excursion(TAPSE),peak systolic velocity of the tricuspid annulus(S`),and partial pressure of carbon dioxide(PaCO2),along with increased rates of emphysema and pulmonary fibrosis(all P<0.05).Compared with LCD-PH,cardiopulmonary-phenotype IPAH patients had higher mPAP,PVR,FEV1%predicted,FEV1/FVC ratio,PAWP,and systolic pulmonary artery pressure,but lower DLCO,cardiac output,TAPSE,S′,and PaCO2(all P<0.05).No statistical difference was observed in PaO2 between these two groups.All cardiopulmonary-phenotype IPAH patients and classical IPAH patients received targeted medications,55%of CLD-PH patients did not receive targeted therapy,45%received monotherapy only(compared with cardiopulmonary-phenotype IPAH patients and classical IPAH groups,P<0.05).The 1,2,and 5-year survival rates were 79.2%,62.2%,and 46.7%,respectively in patients with cardiopulmonary-phenotype IPAH,100%,94.4%,and 94.4%,respectively in classic IPAH patients,and 92.9%,77.4%and 77.4%,respectively in patients with CLD-PH.Survival rates for cardiopulmonary-phenotype IPAH patients were significantly lower than those of classical IPAH and CLD-PH patients(log-rank P=0.008).Conclusions:IPAH cardiopulmonary phenotype patients are older,predominantly male,and often have a smoking history(median 30 pack-years).They exhibit severe hypoxemia,markedly reduced DLCOc,preserved spirometry,and severe pulmonary hypertension and lower survival rate.

Objectives:To investigate the clinical and cardiopulmonary functional characteristics of cardiopulmonary-phenotype idiopathic pulmonary arterial hypertension(IPAH)patients in comparison with classical IPAH patients and pulmonary hypertension patients associated with chronic lung disease(CLD-PH).Methods:In this retrospective study,data were collected from 30 patients with classical IPAH,20 cardiopulmonary phenotype IPAH patients,and 20 patients with CLD-PH,who were hospitalized in the Department of Respiratory and Critical Care Medicine at Beijing Chaoyang Hospital from November 2017 to February 2025.Pulmonary hypertension was diagnosed via right heart catheterization and all patients underwent pulmonary function tests,chest computed tomography(CT),echocardiography,and patients were followed up to 5 years.Results:Compared to classical IPAH patients,both cardiopulmonary phenotype IPAH patients and CLD-PH patients exhibited later onset age,higher proportions of World Health Organization(WHO)functional class Ⅲ-Ⅳ,males,and smokers(all P<0.05).The cardiopulmonary-phenotype IPAH patients also had higher rates of coronary artery disease and diabetes compared to classical IPAH patients(all P<0.05).Physiologically,the cardiopulmonary-phenotype IPAH patients showed reduced diffusing capacity for carbon monoxide(DLCO)and partial pressure of oxygen(PaO2),along with higher rates of emphysema and pulmonary fibrosis compared to classical IPAH(all P<0.05).In contrast,CLD-PH patients had lower mean pulmonary arterial pressure(mPAP),pulmonary vascular resistance(PVR),and pulmonary artery wedge pressure(PAWP),as well as reduced forced expiratory volume in the first second(FEV1%predicted),FEV1/forced vital capacity(FVC)ratio,and DLCO.However,CLD-PH patients demonstrated higher tricuspid annular plane systolic excursion(TAPSE),peak systolic velocity of the tricuspid annulus(S`),and partial pressure of carbon dioxide(PaCO2),along with increased rates of emphysema and pulmonary fibrosis(all P<0.05).Compared with LCD-PH,cardiopulmonary-phenotype IPAH patients had higher mPAP,PVR,FEV1%predicted,FEV1/FVC ratio,PAWP,and systolic pulmonary artery pressure,but lower DLCO,cardiac output,TAPSE,S′,and PaCO2(all P<0.05).No statistical difference was observed in PaO2 between these two groups.All cardiopulmonary-phenotype IPAH patients and classical IPAH patients received targeted medications,55%of CLD-PH patients did not receive targeted therapy,45%received monotherapy only(compared with cardiopulmonary-phenotype IPAH patients and classical IPAH groups,P<0.05).The 1,2,and 5-year survival rates were 79.2%,62.2%,and 46.7%,respectively in patients with cardiopulmonary-phenotype IPAH,100%,94.4%,and 94.4%,respectively in classic IPAH patients,and 92.9%,77.4%and 77.4%,respectively in patients with CLD-PH.Survival rates for cardiopulmonary-phenotype IPAH patients were significantly lower than those of classical IPAH and CLD-PH patients(log-rank P=0.008).Conclusions:IPAH cardiopulmonary phenotype patients are older,predominantly male,and often have a smoking history(median 30 pack-years).They exhibit severe hypoxemia,markedly reduced DLCOc,preserved spirometry,and severe pulmonary hypertension and lower survival rate.

Objective This study examined changes in serum growth hormone(GH)and insulin-like growth factor 1(IGF-1)levels,as well as insulin resistance,in patients with type 2 diabetes mellitus(T2DM)across different stages of diabetic kidney disease(DKD).It explored the relationship between these factors and the occurrence and progression of DKD,with the aim of identifying new targets for the prevention and treatment of DKD.Methods In this cross-sectional study,T2DM patients aged 18-80 years were selected from the Department of Endocrinology,The Second Affiliated Hospital of Xi'an Jiaotong University,from November 30,2019 to March 30,2021.Their serum GH and IGF-1 levels were tested.Results The serum GH level increased gradually in the groups,and the difference was statistically significant(P＜0.05).The level of serum IGF-1 decreased first and then increased among the groups,but with no significant difference(P＞0.05).HOMA-IR showed no significant correlation among the groups(P＞0.05),but HOMA-β gradually decreased among the groups,with statistically significant difference(P＜0.05).Multivariate Logistic regression analysis showed that the increase of serum GH level was closely related to the progression of T2DM to early DKD,and that the increase of serum IGF-1 level might be related to the deterioration of renal function in patients with T2DM at DKD stage.HOMA-IR and HOMA-β were not found to have correlation with the occurrence or development of DKD in T2DM patients.Conclusion In patients with T2DM,elevated serum GH level is closely associated with the occurrence and progression of early DKD.The increase of serum GH level may promote the occurrence of early DKD in T2DM,and the increase of serum IGF-1 level may be related to the deterioration of renal function in clinical DKD stage.In this study,we did not find correlation of HOMA-IR and HOMA-β with the occurrence or development of DKD in T2DM patients.

The COVID-19 epidemic has spread to the whole world for three years and has had a serious impact on human life, health and economic activities. China's epidemic prevention and control has gone through the following stages: emergency unconventional stage, emergency normalization stage, and the transitional stage from the emergency normalization to the "Category B infectious disease treated as Category B" normalization, and achieved a major and decisive victory. The designated hospitals for prevention and control of COVID-19 epidemic in Tianjin has successfully completed its tasks in all stages of epidemic prevention and control, and has accumulated valuable experience. This article summarizes the experience of constructing a hospital infection prevention and control system during the "Category B infectious disease treated as Category A" period in designated hospital. The experience is summarized as the "Cluster" hospital infection prevention and control system, namely "three rings" outside, middle and inside, "three districts" of green, orange and red, "three things" before, during and after the event, "two-day pre-purification" and "two-director system", and "one zone" management. In emergency situations, we adopt a simplified version of the cluster hospital infection prevention and control system. In emergency situations, a simplified version of the "Cluster" hospital infection prevention and control system can be adopted. This system has the following characteristics: firstly, the system emphasizes the characteristics of "cluster" and the overall management of key measures to avoid any shortcomings. The second, it emphasizes the transformation of infection control concepts to maximize the safety of medical services through infection control. The third, it emphasizes the optimization of the process. The prevention and control measures should be comprehensive and focused, while also preventing excessive use. The measures emphasize the use of the least resources to achieve the best infection control effect. The fourth, it emphasizes the quality control work of infection control, pays attention to the importance of the process, and advocates the concept of "system slimming, process fattening". Fifthly, it emphasizes that the future development depends on artificial intelligence, in order to improve the quality and efficiency of prevention and control to the greatest extent. Sixth, hospitals need to strengthen continuous training and retraining. We utilize diverse training methods, including artificial intelligence, to ensure that infection control policies and procedures are simple. We have established an evaluation and feedback mechanism to ensure that medical personnel are in an emergency state at all times.

Objective:To investigate the relationship between intramuscular adipose tissue index (IATI) calculated from computed tomography images at transverse process of the first lumbar and all-cause mortality in maintenance dialysis patients, and to provide a reference for improving the prognosis in these patients.Methods:It was a multicenter retrospective cohort study. The clinical data of patients who received maintenance hemodialysis or peritoneal dialysis treatment from January 1, 2017 to December 31, 2019 in 4 grade Ⅲ hospitals including Zhongda Hospital Affiliated to Southeast University, Taizhou People's Hospital Affiliated to Nanjing Medical University, Affiliated Hospital of Yangzhou University, and the Third Affiliated Hospital of Soochow University were retrospectively collected. IATI was calculated by low attenuation muscle (LAM) density/skeletal muscle density. The receiver-operating characteristic curve was used to determine the optimal cut-off value of IATI, and the patients were divided into high IATI group and low IATI group according to the optimal cut-off value. The differences of baseline clinical data and measurement parameters of the first lumbar level between the two groups were compared. The follow-up ended on December 23, 2022. The endpoint event was defined as all-cause mortality within 3 years. Kaplan-Meier survival curve and log-rank test were used to analyze the survival rates and the differences between the two groups. Multivariate Cox regression analysis models were used to analyze the association between IATI and the risk of all-cause mortality in maintenance dialysis patients. Multivariate logistic regression analysis model was used to analyze the influencing factors of high IATI.Results:A total of 478 patients were eligibly recruited in this study, with age of (53.55±13.19) years old and 319 (66.7%) males, including 365 (76.4%) hemodialysis patients and 113 (23.6%) peritoneal dialysis patients. There were 376 (78.7%) patients in low IATI (<0.42) group and 102 (21.3%) patients in high IATI (≥0.42) group. The proportion of age ≥ 60 years old ( χ2=24.746, P<0.001), proportion of diabetes mellitus ( χ2=5.570, P=0.018), fasting blood glucose ( t=-2.145, P=0.032), LAM density ( t=-3.735, P<0.001), LAM index ( t=-7.072, P<0.001), and LAM area/skeletal muscle area ratio ( Z=-9.630, P<0.001) in high IATI group were all higher than those in low IATI group, while proportion of males ( χ2=11.116, P<0.001), serum albumin ( Z=2.708, P=0.007) and skeletal muscle density ( t=12.380, P<0.001) were lower than those in low IATI group. Kaplan-Meier survival analysis showed that the 3-years overall survival rate of low IATI group was significantly higher than that in high IATI group (Log-rank χ2=19.188, P<0.001). Multivariate Cox regression analysis showed that IATI<0.42 [<0.42/≥0.42, HR(95% CI): 0.50 (0.31-0.83), P=0.007] was an independent protective factor of all-cause mortality, and age ≥60 years old [ HR (95% CI): 2.61 (1.60-4.23), P<0.001], diabetes mellitus [ HR (95% CI): 1.71 (1.06-2.78), P=0.029] and high blood neutrophil/lymphocyte ratio [ HR (95% CI): 1.04 (1.00-1.07), P=0.049] were the independent risk factors of all-cause mortality in maintenance dialysis patients. Stepwise Cox regression analysis showed that IATI<0.42 was still an independent protective factor of all-cause mortality in maintenance dialysis patients [<0.42/≥0.42, HR (95% CI): 0.45 (0.27-0.76), P=0.003]. Multivariate logistic regression analysis showed that low skeletal muscle density [ OR (95% CI): 0.84 (0.81-0.88), P<0.001] and high serum triglyceride [ OR (95% CI): 1.39 (1.07-1.82), P=0.015] were the independent influencing factors of IATI≥0.42. Conclusion:IATI<0.42 of the first lumbar level is an independent protective factor of all-cause mortality in maintenance dialysis patients. Localized myosteatosis within high-quality skeletal muscle may reduce the risk of all-cause mortality in these patients.

Bluetongue virus is an arbovirus that seriously harms ruminants such as sheep,this study aims to investigate the molecular mechanism of bluetongue virus infection and host cell interferon antiviral immune response.The study was conducted to characterize the mRNA expression of inter-feron pathway genes by real-time fluorescence quantitative PCR,as well as Western blot analysis of MDA5,TRAF3,RIG-Ⅰ,and TBK1 protein expression in BHK-21 cells induced by BTV with a multiplicity of infections(MOI)of 1 for 18,24,and 36 h.The results showed that the most pro-nounced changes in the expression of interferon signaling pathway genes were observed at 24 h of induction,the gene mRNA expression levels of the IFN-α,IFN-β,RIG-Ⅰ,TBK1,MDA5,VISA,and TRAF3 genes were upregulated.However,the mRNA expression levels of IKKε and TRAF6 genes were downregulated.At the protein level,MDA5 and TBK1 proteins were upregulated while RIG-1 and TRAF3 proteins were downregulated,which showed that BTV infection induces a typeⅠ interferon immune response in BHK-21 cells.This study lays the foundation for further exploring the antiviral immunity mechanism of IFN-Ⅰ signaling pathway regulatory genes in host cells infected with BTV infection.

Objective:To determine the prevalence and risk factors of constipation in Hangzhou urban areas by analyzing data from a recent epidemiological survey.Methods:From August 2022 to June 2023, a cross-sectional survey was conducted on 43 communities on eight streets in suburbs of Hangzhou, including Wangjiang, Sijiqing, Puyan, Changhe, Ziyang, Xiaoshanchengxiang, Zhuantang, and Banshan. Written questionnaires were filled out face to face by community doctors. The prevalence of constipation was investigated using a combination of the Bristol stool typing method and the Rome IV criteria. The risk factors for constipation were analyzed using multivariate logistic regression analysis.Results:The study cohort comprised 10,479 participants, 5551 of whom were male (53.0%) and 4928 female (47.0%). The overall prevalence of constipation was 10.6% (1107/10,479). The prevalence was 11.0% (613/5551) for male and 10.0% (494/4928) for female participants; this difference is not statistically significant ( P>0.05).There were no significant differences in the prevalence of constipation between different categories of body mass index ( P>0.05). The prevalence of constipation was highest in individuals aged 70 years and above, accounting for 15.4% (264/1717) of all cases. There were significant differences in the prevalence of constipation between different occupations, marital statuses, and educational levels ( P<0.001). Patients with hypertension or diabetes mellitus had a higher prevalence of constipation than those without these conditions, those who had been taking long-term medication had a higher prevalence of constipation than those who did not, parents of children with constipation had a higher prevalence of constipation than those with children who did not have constipation, and there were other significant differences in various variables ( P<0.001). Multivariate logistic regression analysis of modifiable factors revealed that eating less fruit (OR=1.284, 95%CI: 1.061–1.553, P=0.010), liking spicy and salty food (OR=1.234, 95%CI: 1.039–1.466, P=0.016), sleeping for too long (OR=1.644, 95%CI: 1.260–2.144, P<0.001), irregular sleep patterns (OR=1.370, 95%CI: 1.127–1.665, P=0.002) and minimal exercise (OR=1.388, 95% CI: 1.168–1.649, P<0.001) were all risk factors for constipation; whereas mix diets (OR=0.709,95%CI:0.608–0.826, P<0.001), meat diets (OR=0.604,95%CI: 0.495–0.736, P<0.001), moderate alcohol consumption (OR=0.659, 95% CI: 0.534–0.812, P<0.001), less or no drinking (OR=0.523, 95% CI: 0.428–0.638, P<0.001), and non-smoking (OR=0.819, 95%CI:0.674–0.995, P<0.001) protected against constipation. We found no association between dietary preferences and the risk of constipation (refined grains: OR=1.147, 95%CI:0.944–1.393, P=0.167; no specific preference: OR=0.935, 95%CI:0.783–1.117, P=0.459). The rate of agreement between self-evaluation of constipation by the respondents and objective criteria was higher for negatives (95.8%) than for positives (38.0%). Conclusion:The prevalence of constipation is relatively high in the urban population of Hangzhou. A good lifestyle and diet can significantly reduce its incidence. There should be more emphasis on preventive education, promotion of prevention of constipation, and advocacy for a healthy lifestyle.