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.

Apical microsurgery is accurate and minimally invasive, produces few complications, and has a success rate of more than 90%. However, due to the lack of awareness and understanding of apical microsurgery by dental general practitioners and even endodontists, many clinical problems remain to be overcome. The consensus has gathered well-known domestic experts to hold a series of special discussions and reached the consensus. This document specifies the indications, contraindications, preoperative preparations, operational procedures, complication prevention measures, and efficacy evaluation of apical microsurgery and is applicable to dentists who perform apical microsurgery after systematic training.
Microsurgery/standards* ; Humans ; Apicoectomy ; Contraindications, Procedure ; Tooth Apex/diagnostic imaging* ; Postoperative Complications/prevention & control* ; Consensus ; Treatment Outcome

Objective To investigate the epidemiological status and molecular characteristics of diarrheagenic Escherichia coli(DEC)infection in Xiangshan County,providing a basis for its infection surveillance and outbreak control.Methods The real-time fluores-cence quantitative PCR(qRT-PCR)was used to detect virulence genes in isolated E.coli strains.The identified DEC strains were per-formed whole genome sequencing(WGS).The sequencing data were utilized for bacterial resistance gene analysis,multilocus sequence typing(MLST),and core genome multilocus sequence typing(cgMLST).Results A total of 29 DEC strains were isolated,with a detection rate of 9.8%.Enteroaggregative E.coli(EAEC)and enterotoxigenic E.coli(ETEC)were the main strains,accounting for 51.7%and 31.1%,respectively,while enteropathogenic E.coli(EPEC)had a relatively low proportion(17.2%).Drug resistance gene analysis showed that these strains were generally resistant to multiple antibiotics such as fluoroquinolones and carbapenems.MLST typing identified 19 sequence types(STs),including one novel ST type(STcpEC1).Among them,ST117,ST2178,ST31,and ST48 were dominant types.ETEC strains were mainly concentrated in the clonal complex CC-10,while the molecular typing of EAEC and EPEC strains was more dispersed.cgMLST analysis further confirmed the genetic characteristics of different DEC strains.Conclusion DEC infections in Xiangshan County are mainly caused by EAEC,ETEC,and EPEC.These strains have abundant resistance genes.Among them,the molecular typing of EAEC and EPEC is relatively dispersed,while ETEC is concentrated in the clonal complex CC-10.

AIM:This study aims to investigate the expression and prognostic value of ubiquitin-conjugating enzyme E2C(UBE2C)in hepatocellular carcinoma(HCC),and to explore its impact on cancer stemness and the regulato-ry mechanisms.METHODS:(1)The TCGA and GEO databases were used to analyze UBE2C mRNA expression levels in HCC tissues and their correlation with prognosis using bioinformatics techniques,and these findings were further vali-dated in postoperative specimens from 107 HCC patients.The GEPIA database was used to analyze the correlation be-tween UBE2C and steroid receptor coactivator(SRC).(2)The CCK8,colony formation,wound healing,and spheroid formation assays were used to assess the effects of UBE2C on HCC cell proliferation,migration,and stemness.The inter-action between UBE2C and signal transducer and activator of transcription 3(STAT3),as well as its regulatory mecha-nism,was examined by Western blot and co-immunoprecipitation assays.(3)The subcutaneous xenograft model in nude mice was employed to validate the role of UBE2C in tumor growth in vivo.RESULTS:(1)Bioinformatics analysis re-vealed that UBE2C expression was significantly up-regulated in HCC tissues compared with adjacent normal tissues(P<0.05 in TCGA,and P<0.01 in GEO).Consistently,analysis of HCC specimens confirmed that high UBE2C expression was associated with shortened overall survival and disease-free survival of HCC patients(P<0.05).Furthermore,analysis using the GEPIA database revealed a positive correlation between UBE2C and SRC(P<0.01).(2)In vitro experiments demonstrated that UBE2C significantly promotes the proliferation and migration of HCC cells.Based on these findings,we presumed that UBE2C may regulate the phosphorylation of STAT3 at the Y705 site by modulating SRC activity,thereby in-fluencing the stemness characteristics of tumor cells.(3)In vivo experiments further confirmed that UBE2C inhibition sig-nificantly suppressed tumor growth.CONCLUSION:The UBE2C promoted proliferation and migration of HCC cells and regulated the stemness of HCC cells by interacting with STAT3.

Objective:To summarize the best evidence for oral nutrition management of patients with inflammatory bowel disease (IBD) and to evaluate its effectiveness.Methods:From May 2023 to March 2024, IBD patients, the medical and nursing staff in the Department of Proctology, Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine, were selected as study subjects using convenience sampling method. Evidence-based nursing was used to summarize the best evidence for oral nutrition management of IBD patients, formulate review indicators, develop best evidence-based practice programs for clinical application through status review and barrier factor analysis. Effectiveness was evaluated at the system level, medical and nursing staff level, and patient level.Results:After evidence-based practice, a multidisciplinary management team and management system for oral nutrition was established at the system level. At the medical and nursing staff level, the oral nutrition knowledge score increased from [50.00 (40.00, 60.00) ] to [100.00 (85.00, 100.00) ], with a statistically significant difference ( P<0.01), and the implementation rate of the three review indicators improved from 0 to 100.00%. At the patient level, oral nutrition knowledge and attitude scores improved from [10.00 (8.00, 12.00) ] and [35.00 (31.00, 40.00) ] to [16.00 (16.00, 16.00) ] and [48.00 (45.00, 48.00) ], respectively, with statistically significant differences ( P<0.01), and the implementation rate of each of the review indicators rose above 80.00% except for indicator 6. The patients' nutrition indicators, including body mass index, waist circumference, left upper arm midpoint circumference and left hand grip strength were improved, and the difference was statistically significant ( P<0.05) . Conclusions:The implementation of evidence-based practice programs can improve the standardization of clinical work of healthcare professionals, enhance patients' nutrition knowledge and attitude, and promote their rational dietary behaviors, thus improving their nutrition status and quality of life.

Objective To investigate the epidemiological status and molecular characteristics of diarrheagenic Escherichia coli(DEC)infection in Xiangshan County,providing a basis for its infection surveillance and outbreak control.Methods The real-time fluores-cence quantitative PCR(qRT-PCR)was used to detect virulence genes in isolated E.coli strains.The identified DEC strains were per-formed whole genome sequencing(WGS).The sequencing data were utilized for bacterial resistance gene analysis,multilocus sequence typing(MLST),and core genome multilocus sequence typing(cgMLST).Results A total of 29 DEC strains were isolated,with a detection rate of 9.8%.Enteroaggregative E.coli(EAEC)and enterotoxigenic E.coli(ETEC)were the main strains,accounting for 51.7%and 31.1%,respectively,while enteropathogenic E.coli(EPEC)had a relatively low proportion(17.2%).Drug resistance gene analysis showed that these strains were generally resistant to multiple antibiotics such as fluoroquinolones and carbapenems.MLST typing identified 19 sequence types(STs),including one novel ST type(STcpEC1).Among them,ST117,ST2178,ST31,and ST48 were dominant types.ETEC strains were mainly concentrated in the clonal complex CC-10,while the molecular typing of EAEC and EPEC strains was more dispersed.cgMLST analysis further confirmed the genetic characteristics of different DEC strains.Conclusion DEC infections in Xiangshan County are mainly caused by EAEC,ETEC,and EPEC.These strains have abundant resistance genes.Among them,the molecular typing of EAEC and EPEC is relatively dispersed,while ETEC is concentrated in the clonal complex CC-10.

AIM:This study aims to investigate the expression and prognostic value of ubiquitin-conjugating enzyme E2C(UBE2C)in hepatocellular carcinoma(HCC),and to explore its impact on cancer stemness and the regulato-ry mechanisms.METHODS:(1)The TCGA and GEO databases were used to analyze UBE2C mRNA expression levels in HCC tissues and their correlation with prognosis using bioinformatics techniques,and these findings were further vali-dated in postoperative specimens from 107 HCC patients.The GEPIA database was used to analyze the correlation be-tween UBE2C and steroid receptor coactivator(SRC).(2)The CCK8,colony formation,wound healing,and spheroid formation assays were used to assess the effects of UBE2C on HCC cell proliferation,migration,and stemness.The inter-action between UBE2C and signal transducer and activator of transcription 3(STAT3),as well as its regulatory mecha-nism,was examined by Western blot and co-immunoprecipitation assays.(3)The subcutaneous xenograft model in nude mice was employed to validate the role of UBE2C in tumor growth in vivo.RESULTS:(1)Bioinformatics analysis re-vealed that UBE2C expression was significantly up-regulated in HCC tissues compared with adjacent normal tissues(P<0.05 in TCGA,and P<0.01 in GEO).Consistently,analysis of HCC specimens confirmed that high UBE2C expression was associated with shortened overall survival and disease-free survival of HCC patients(P<0.05).Furthermore,analysis using the GEPIA database revealed a positive correlation between UBE2C and SRC(P<0.01).(2)In vitro experiments demonstrated that UBE2C significantly promotes the proliferation and migration of HCC cells.Based on these findings,we presumed that UBE2C may regulate the phosphorylation of STAT3 at the Y705 site by modulating SRC activity,thereby in-fluencing the stemness characteristics of tumor cells.(3)In vivo experiments further confirmed that UBE2C inhibition sig-nificantly suppressed tumor growth.CONCLUSION:The UBE2C promoted proliferation and migration of HCC cells and regulated the stemness of HCC cells by interacting with STAT3.

Objective:To summarize the best evidence for oral nutrition management of patients with inflammatory bowel disease (IBD) and to evaluate its effectiveness.Methods:From May 2023 to March 2024, IBD patients, the medical and nursing staff in the Department of Proctology, Zhongshan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine, were selected as study subjects using convenience sampling method. Evidence-based nursing was used to summarize the best evidence for oral nutrition management of IBD patients, formulate review indicators, develop best evidence-based practice programs for clinical application through status review and barrier factor analysis. Effectiveness was evaluated at the system level, medical and nursing staff level, and patient level.Results:After evidence-based practice, a multidisciplinary management team and management system for oral nutrition was established at the system level. At the medical and nursing staff level, the oral nutrition knowledge score increased from [50.00 (40.00, 60.00) ] to [100.00 (85.00, 100.00) ], with a statistically significant difference ( P<0.01), and the implementation rate of the three review indicators improved from 0 to 100.00%. At the patient level, oral nutrition knowledge and attitude scores improved from [10.00 (8.00, 12.00) ] and [35.00 (31.00, 40.00) ] to [16.00 (16.00, 16.00) ] and [48.00 (45.00, 48.00) ], respectively, with statistically significant differences ( P<0.01), and the implementation rate of each of the review indicators rose above 80.00% except for indicator 6. The patients' nutrition indicators, including body mass index, waist circumference, left upper arm midpoint circumference and left hand grip strength were improved, and the difference was statistically significant ( P<0.05) . Conclusions:The implementation of evidence-based practice programs can improve the standardization of clinical work of healthcare professionals, enhance patients' nutrition knowledge and attitude, and promote their rational dietary behaviors, thus improving their nutrition status and quality of life.

Objective To investigate the characteristics of mismatch negativity(MMN)in patients with total deafness of sudden hearing loss with different prognosis,and to assess whether MMN is associated with the progno-sis of sudden hearing loss patients.Methods Fifty-one cases of sudden hearing loss with unilateral total deafness were selected for pure tone audiometry(PTA)and auditory MMN examination before treatment,and 26 subjects with mornal hearing were included as the normal group.After routine treatment for 10 days according to the guide-lines for sudden hearing loss,PT A examination was performed to determine their recovery status,and they were di-vided into ineffective group and effective group according to prognosis.The characteristics of MMN latency and am-plitude in the effective group and the ineffective group were observed and compared with the normal control group.Results The MMN waveform was elicited normally in 51 patients with sudden hearing loss and 26 cases in the nor-mal group,of which 29 were in effective group and 22 were in ineffective group.There were significant statistical differences in MMN latency between the patients group and the normal group(P＜0.05).Anova showed no signifi-cant difference in MMN latency between the invalid group and the effective group(P＞0.05),and there was no sig-nificant difference in amplitude among the three groups(P＞0.05).Conclusion The MMN latency of sudden hear-ing loss patients was signifanty shortened.There were no differences in MMN latency and amplitude between the in-effective group and the effective group.