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 investigate the anti-inflammatory effect of electroacupuncture in mice with chronic obstructive pul-monary disease(COPD)and its underlying mechanisms.METHODS Forty C57BL/6 mice were randomly divided into normal group,model group,electroacupuncture(EA)group,vagotomy group,and vagotomy+EA group,with 8 mice in each group.Except for the normal group,all groups were exposed to cigarette smoke for 12 weeks to establish a COPD model.After model establishment,the vagotomy group and the vagotomy+EA group underwent left cervical vagotomy before EA.EA treatment was performed at the Feishu(BL13)and Zusanli(ST36)acupoints once daily for 20 minutes for a total of 14 days.After EA,the pulmonary ventilation function of the mice was detected by a pulmonary function analysis system;lung tissue pathology was observed by HE staining;the levels of inter-leukin(IL)-6,tumor necrosis factor-α(TNF-α),IL-10,and transforming growth factor-β(TGF-β)in lung tissue were detected by ELISA;the expression of CD86 and CD206 proteins in lung tissue was detected by Western blot;the distribution of F4/80+CD86+(M1 type)and F4/80+CD206+(M2 type)cells in lung tissue was determined by flow cytometry;the expression of CD86 and CD206 in lung tissue was observed by immunofluorescence.RESULTS Compared with the normal group,the model group showed significantly decreased lung function(P<0.01),obvious lung pathological damage,increased M1 proportion,IL-6,TNF-α,CD86 content and expression(P<0.05,P<0.01),and decreased M2 proportion,IL-10,TGF-β,CD206 content and expression(P<0.01).Compared with the model group,the EA group showed varying degrees of improvement in lung function and pathology;the M1 proportion,IL-6,TNF-α,and CD86 were reduced(P<0.05,P<0.01),while the M2 proportion,IL-10,TGF-β,and CD206 were increased(P<0.05,P<0.01).The vagotomy group showed worsened lung function and pathology,increased IL-6,TNF-α,and CD86 content and expression(P<0.05,P<0.01),and decreased IL-10,TGF-β,and CD206 content and expression(P<0.05,P<0.01).Compared with the EA group,the vagotomy+EA group showed increased M1 proportion,IL-6,TNF-α,and CD86 content and expression(P<0.01),and decreased M2 proportion,IL-10,TGF-β,and CD206 content and expression(P<0.05,P<0.01).CONCLUSION EA at Feishu(BL13)and Zusanli(ST36)acupoints can improve lung function and pulmonary inflammation in COPD mice,promoting the polarization of alveolar macrophages from M1 to M2,which is mediated by the vagus nerve.

Objective:To investigate the mediating effect of work engagement on the relationship between nurses′ perception of authentic leadership and their innovative behavior, providing insights for enhancing nurses′ innovative practices.Methods:A convenience sampling method was employed to recruit registered nurses from 37 hospitals in Anhui Province between January and March 2022. The Authentic Leadership Questionnaire, Utrecht Work Engagement Scale and Nurse Innovative Behavior Scale were utilized to conduct a cross-sectional survey. SPSS 25.0 was used to analyze the correlations among nurses′ perception of authentic leadership, work engagement, and innovative behavior, while AMOS 26.0 was employed to assess the mediating effect of work engagement between nurses′ perception of authentic leadership and innovative behavior.Results:A total of 3 200 questionnaires were distributed, and 2 994 valid questionnaires were collected. Among the 2 994 participating nurses, 2 907 (97.1%) were female and 87 (2.9%) were male, 1 002 (33.5%) aged 22-25 years, 1 374 (45.9%) aged 26-39 years, 467 (15.6%) aged 40-49 years, and 151 (5.0%) aged 50 years or older. The total scores of the Authentic Leadership Questionnaire, Utrecht Work Engagement Scale and Nurse Innovative Behavior Scale were (68.23 ± 13.15), (51.49 ± 11.65) and (38.28 ± 8.35) points, respectively. Pearson correlation analysis revealed significant positive correlations between nurses′ perception of authentic leadership and work engagement ( r=0.473, P<0.01), between nurses′ perception of authentic leadership and innovative behavior ( r=0.530, P<0.01), and between work engagement and innovative behavior ( r=0.553, P<0.01). The mediating effect analysis indicated that work engagement partially mediates the relationship between nurses′ perception of authentic leadership and innovative behavior ( β=0.18, P<0.01), with a mediation effect ratio of 34.62%. Conclusions:Nurses′ perception of authentic leadership can indirectly influence their innovative behavior through work engagement. It is recommended that nursing managers prioritize the enhancement of authentic leadership and foster work engagement among nurses to stimulate their innovative behavior.

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.

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.

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 To investigate the effects and underlying mechanisms of Yiqi Huoxue Decoction(YQHX)on mitochon-drial midzone division and peripheral fission in myocardial tissue after myocardial infarction(MI).METHODS A total of 48 male SPF-grade C57BL/6N mice were randomly divided into a sham-operated group(Sham,n=12)and a left anterior descending coronary ar-tery ligation MI model(n=36).After MI surgery,mice deemed to have successfully developed the model were randomly divided into a model group(MI,n=12),a YQHX group(n=12),and an empagliflozin group(EMPA,n=12)based on echocardiographic results.Four weeks after infarction,cardiac function and structural changes were comprehensively evaluated using echocardiography imaging,serum myocardial injury biomarkers,and hematoxylin-eosin(HE)staining.Transmission electron microscopy(TEM)was employed to observe mitochondrial ultrastructural,morphological,and quantitative changes at the peri-infarct zone.Myocardial mitochondria and cytoplas-mic fractions were isolated from myocardial tissue using a mitochondrial extraction kit,and the spatial expression changes of mitochon-drial fission-related proteins in both mitochondria and cytoplasm of the peri-infarct myocardium were analyzed by Western blot.These proteins included dynamin-related protein 1(Drp1),its phosphorylated form at serine 616(P-Drp1-Ser616),mitochondrial fission fac-tor(MFF),and mitochondrial fission protein 1(Fis1).RESULTS Compared with the MI group,mice in the YQHX group exhibited sig-nificantly increased left ventricular ejection fraction(LVEF)and left ventricular fractional shortening(LVFS)(P<0.000 1),as well as decreased left ventricular internal dimension-diastole(LVIDd)and left ventricular end-systolic diameter(LVIDs)(P<0.05,P<0.01),suggesting improved cardiac function.Additionally,serum levels of lactate dehydrogenase(LDH)and creatine kinase-MB(CK-MB)were significantly reduced in the YQHX group(P<0.05,P<0.001),indicating cardio-protective effects of YQHX against ischemic in-jury.HE staining showed that YQHX improved cellular morphology,suggesting structural improvement.TEM showed that YQHX sig-nificantly improved mitochondrial swelling and reduced mitochondrial fragmentation in the marginal zone of myocardial infarction,thereby preserving mitochondrial ultrastructure.Furthermore,Western blot showed that YQHX treatment significantly downregulated P-Drp1-Ser616 expression(P<0.05)in the cytoplasm.Interestingly,YQHX treatment significantly downregulated mitochondrial Fis1 expression(P<0.05),thereby inhibiting peripheral mitochondrial fission.Meanwhile,YQHX treatment significantly increased MFF ex-pression in mitochondria(P<0.01),which may promote mitochondrial midzone fission.CONCLUSION YQHX improves cardiac structure and function after MI,potentially by promoting myocardial mitochondrial midzone fission and inhibiting mitochondrial periph-eral fission in ischemic cardiomyocytes.

OBJECTIVE To investigate the anti-inflammatory effect of electroacupuncture in mice with chronic obstructive pul-monary disease(COPD)and its underlying mechanisms.METHODS Forty C57BL/6 mice were randomly divided into normal group,model group,electroacupuncture(EA)group,vagotomy group,and vagotomy+EA group,with 8 mice in each group.Except for the normal group,all groups were exposed to cigarette smoke for 12 weeks to establish a COPD model.After model establishment,the vagotomy group and the vagotomy+EA group underwent left cervical vagotomy before EA.EA treatment was performed at the Feishu(BL13)and Zusanli(ST36)acupoints once daily for 20 minutes for a total of 14 days.After EA,the pulmonary ventilation function of the mice was detected by a pulmonary function analysis system;lung tissue pathology was observed by HE staining;the levels of inter-leukin(IL)-6,tumor necrosis factor-α(TNF-α),IL-10,and transforming growth factor-β(TGF-β)in lung tissue were detected by ELISA;the expression of CD86 and CD206 proteins in lung tissue was detected by Western blot;the distribution of F4/80+CD86+(M1 type)and F4/80+CD206+(M2 type)cells in lung tissue was determined by flow cytometry;the expression of CD86 and CD206 in lung tissue was observed by immunofluorescence.RESULTS Compared with the normal group,the model group showed significantly decreased lung function(P<0.01),obvious lung pathological damage,increased M1 proportion,IL-6,TNF-α,CD86 content and expression(P<0.05,P<0.01),and decreased M2 proportion,IL-10,TGF-β,CD206 content and expression(P<0.01).Compared with the model group,the EA group showed varying degrees of improvement in lung function and pathology;the M1 proportion,IL-6,TNF-α,and CD86 were reduced(P<0.05,P<0.01),while the M2 proportion,IL-10,TGF-β,and CD206 were increased(P<0.05,P<0.01).The vagotomy group showed worsened lung function and pathology,increased IL-6,TNF-α,and CD86 content and expression(P<0.05,P<0.01),and decreased IL-10,TGF-β,and CD206 content and expression(P<0.05,P<0.01).Compared with the EA group,the vagotomy+EA group showed increased M1 proportion,IL-6,TNF-α,and CD86 content and expression(P<0.01),and decreased M2 proportion,IL-10,TGF-β,and CD206 content and expression(P<0.05,P<0.01).CONCLUSION EA at Feishu(BL13)and Zusanli(ST36)acupoints can improve lung function and pulmonary inflammation in COPD mice,promoting the polarization of alveolar macrophages from M1 to M2,which is mediated by the vagus nerve.

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.