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 To explore the efficacy of platelet-rich plasma （PRP） intrauterine infusion combined with Gushen antai pills in the treatment of recurrent abortion （RSA） and its impacts on endometrial receptivity and hormone levels in patients. METHODS A total of 108 patients with RSA treated in our hospital from January 2021 to January 2024 were selected and evenly divided into control group and study group using the random number table method， with 54 cases in each group. On the basis of conventional treatment， patients in the control group were administered with Gushen antai pills， while patients in the study group received PRP intrauterine infusion combined with Gushen antai pills. The clinical efficacy， TCM syndrome scores before and after treatment， endometrial receptivity [endometrial thickness （EST）， spiral artery resistance index （RI）， and pulsatility index （PI）]， hormone [progesterone （P）， estradiol （E2）， and human chorionic gonadotropin （HCG）] levels， as well as pregnancy outcomes， were compared between the two groups. Additionally， the occurrence of adverse reactions in both groups was recorded. RESULTS The total effective rate （91.44% vs. 81.48%） and infant live birth rate （96.30% vs. 83.33%） of the study group were significantly higher than those of the control group （P＜0.05）. Following treatment， various TCM syndrome scores and total score， spiral artery RI and PI levels in both groups were markedly lower than those in the same groups before treatment， with the study group showing significantly lower levels than the control group（P＜0.05）. Conversely， the P， E2， HCG and EST levels in both groups were significantly higher than those in the same groups before treatment， and the study group exhibited notably higher levels than the control group （P＜0.05）. There was no statistically significant difference in the total incidence of adverse reactions between the two groups （P＞0.05）. CONCLUSIONS PRP intrauterine infusion combined with Gushen antai pills has good clinical efficacy in the treatment of RSA. It can improve TCM syndromes， enhance endometrial receptivity， regulate hormone levels and improve pregnancy outcomes， and it is highly safe.

Fungal infection is an increasingly serious clinical problem,especially in immunocompromised patients.Fungi,such as Candida albicans,Aspergillus,and Cryptococcus,can form biofilm,which enhance their drug re-sistance and immune evasion ability,thus become major obstacles to the treatment of infection.Biofilm formation is not limited to human body,it can also form on the surface of medical devices,leading to chronic and recurrent infec-tion.This article elaborates the biofilm formation mechanisms and hazards caused by different fungi such as Candi-da albicans and Aspergillus,emphasizes immune suppression,chronic diseases,and medical devices as high-risk factors for biofilm formation.Fungi can form biofilm through multiple stages such as adhesion,proliferation,extra-cellular matrix(ECM)construction,and cell dispersion,and enhance drug resistance based on physical barriers and gene regulation.Although existing antifungal agents are effective in infection caused by planktonic fungi,their treatment efficacy on fungal biofilm is limited.Therefore,the article explores new treatment strategies,including non-pharmacological therapies such as photodynamic therapy and electric field therapy,as well as targeted gene edi-ting and the application of new biomaterials.It is necessary to strengthen the combination of basic research and clini-cal applications,develop efficient and low toxicity treatment scheme to improve the successful treatment rate for patients.

Objective Prepubertal mice are administered subcutaneously with letrozole sustained-release pellets behind the neck and treated with a high-fat diet to establish a mouse model of polycystic ovary syndrome(PCOS).The liver transcriptomes of the model mice are compared with those of the placebo control mice to investigate the underlying mechanisms of liver involvement in the pathogenesis of PCOS.Methods A customized 2 mg dose of letrozole sustained-release pellets with a 40-day release period was used.The control placebo and letrozole pellets were implanted subcutaneously in the dorsal cervical region of 3-4-week-old C57BL/6J mice(8 mice per group)to establish the control group and letrozole-induced PCOS model group.Both groups were treated with a high-fat diet starting the day after administration.The modeling period lasted for 5 weeks,during which body weight and 24-hour food intake were monitored in each group every week.When samples were collected,liver weight was recorded.Pathological changes in ovarian and hepatic tissues were examined by hematoxylin-eosin(HE)staining,while hepatic lipid deposition was observed by Oil Red O staining.The extent of macrophage infiltration in the liver was evaluated via F4/80 immunohistochemical staining,and hepatic fibrosis levels were observed by Masson's trichrome staining.Transcriptomic sequencing was performed to analyze differentially expressed genes(DEGs)in liver tissues between the control and model groups,followed by enrichment analysis of significant DEGs.Quantitative real-time fluorescent quantitative PCR(qPCR)was subsequently used to validate the expression of significant DEGs in liver tissues of both groups.Results Compared with the control group,the model group which received subcutaneous letrozole sustained-release pellets combined with a high-fat diet exhibited significantly increased body weight(P＜0.001),prominent polycystic ovarian morphology,and significantly decreased liver-to-body weight ratio(P＜0.05).However,no significant changes were observed in absolute liver weight(P＞0.05),hepatic histomorphology,or lipid deposition.Transcriptome sequencing identified 119 upregulated and 217 downregulated DEGs in the liver tissues of letrozole-treated mice,which were predominantly enriched in pathways related to cholesterol and steroid biosynthesis,steroid hormone metabolism,and inflammatory responses.qPCR validation demonstrated that mRNA expression of HSD3B2 and HMGCR was significantly upregulated in liver(P＜0.01),while mRNA expression of IL4,CCL2 and COL1A1 was downregulated(P＜0.05)in the model group compared with the control group.However,Masson's trichrome staining and F4/80 immunohistochemical analysis showed no significant changes in hepatic fibrosis or macrophage infiltration.Conclusion Subcutaneous administration of letrozole sustained-release pellets combined with a high-fat diet successfully establishes a mouse model of PCOS.The model mice exhibited significant changes in hepatic gene expression.Liver may contribute to PCOS pathogenesis through regulating cholesterol and steroid metabolism.

Objectives:Financial pressure could have severe consequences for research hospitals.Studying the financial statement information and management measures of"top"hospitals in the US.can provide references for research hospitals in China to ensure long-term financial sustainability and improve competitiveness.Methods:The latest audited financial statements of Mayo Clinic,Cleveland Clinic,The Johns Hopkins Health System,and Mass General Brigham in the United States were collected,descriptive statistical analysis was employed.Results:The"top"four medical institution have a good asset composition,moderate debt ratios,large investment,with net assets continue to increase.They have diversified sources of revenue,with specialists being its key asset,human resource expense accounting for 57％～58％of total expenses,pursuing technological leadership.When facing financial pressure,the first choice is to improve operational efficiency and continue to reinvest in core missions.Conclusion:The practice of four"top"research medical institutions in the US.has provide new perspective for research hospitals in China to ensure long-term financial sustainability and improve market competitiveness.

Objective:To observe whether electroacupuncture(EA)can inhibit activation of JAK1/STAT3 signaling pathway and improve pulmonary inflammation in chronic obstructive pulmonary disease(COPD)by increasing expression of SOCS3.Methods:Atotal of 60 mice were randomly divided into normal group,model(COPD)group,COPD+EA group,COPD+si-SOCS3 group,COPD+si-SOCS3 NC group,COPD+si-SOCS3+EA group,with 10 mice in each group.Mice COPD model was replicated by simple cigarette smoking for three months.After modeling,SOCS3 siRNA was administered to lungs of mice in COPD+si-SOCS3 group and COPD+si-SOCS3+EA group,SOCS3 siRNA NC was administered to COPD+si-SOCS3 NC group.24 h after the first SOCS3 siRNA ad-ministration,mice in COPD+EA and COPD+si-SOCS3+EA groups were treated with EA in"Feishu"and"Zusanli",once every other day,30 min/times for 14 days.Lung function of mice in each group was detected;lung pathological changes were observed by HE staining;IL-6,IL-1β and TNF-α levels in mice broncho alveolar lavage fluid were detected by ELISA;protein expressions of SOCS3,JAK1,STAT3,p-JAK1 and p-STAT3 were detected by Western blot;SOCS3,JAK1 and STAT3 mRNA were detected by RT-PCR.Results:Compared with normal group,mice in COPD group showed decreased lung function,thickened alveolar walls,congestion and edema between tissues,significantly increased levels of inflammatory cytokines IL-6,TNF-α and IL-1β,decreased SOCS3 protein expres-sion,p-JAK1/JAK1 and p-STAT3/STAT3 significantly increased,JAK1 and STAT3 mRNA increased,SOCS3 mRNA decreased(P<0.05).Compared with COPD group,the above indexes of COPD+EA group were improved(P<0.05),and the above indexes of COPD+si-SOCS3 group were more serious(P<0.05),and COPD+si-SOCS3+EA group was improved compared with COPD+si-SOCS3 group(P<0.05).Conclusion:EA can inhibit overactivation of JAK1/STAT3 signaling pathway by up-regulating SOCS3 expression,and thus improve pulmonary inflammation in COPD.

Objective:Under the background of the National Health Commission's recent notice to focus on preventing and resolving risks in the economic operation of public hospitals,guaranteeing the healthy and sustainable operation of public hospitals,and the need for localities to pay close attention to the key risks of public hospitals'failure to meet expenditures and revenues,long-term debt,and salary arrearage,the study of the methods used by the Mayo Clinic in the US to maintain solid growth in its financial performance over the long term may serve as a lesson for the high-quality development of China's research-based hospitals.Methods:Mayo Clinic's audited consolidated financial annual reports and related information for the years 2021-2022 and 2022-2023 were collected,and descriptive statistics were analyzed for the financial statements for the years 2021-2023.Results:Mayo Clinic's overall financial position,results of operations,and ability to generate cash from operations for 2021-2023 are strong.Conclusion:The composition of diversified revenue sources helps Mayo Clinic's financial performance to sustain solid growth.This is driven by the values of putting patients'needs and interests first,bringing together a large pool of talented individuals,and adopting a teamwork approach to efficiently deliver a quality experience that exceeds patients'expectations.It is recommended that Chinese research hospitals emphasize international healthcare,patent transfer and investment,and community donations to diversify their revenue streams,which will help to maintain robust financial performance.

During the 14th Five-Year Plan period, the population of China has turned into a stage of negative growth, the fertility rate gradually declined, and the infertility rate gradually increased. The most effective treatment for infertility is assisted reproductive technology (ART), which has brought hope for fertility to many patients. However, the current promotion of the utilization of ART is still fraught with many obstacles. In this context, focusing on the realistic challenges of ART on its development in China, we will take a problem-oriented approach, utilize the relevant policies of ART in foreign countries as references, and put forward policy recommendations for the further implementation of ART, such as coordinating the balanced distribution of ART resources in the region, strengthening policy safeguards for ART, enhancing the institutional functioning of professional associations in medical ethics, and standardizing the application of ART treatments.