Objective:To investigate the clinicopathological features, genetic characteristics, and differential diagnosis of glomangiomatosis with uncertain malignant potential.Methods:Two cases of glomangiomatosis with uncertain malignant potential were collected at Henan Provincial People′s Hospital from 2013 and 2023. Immunohistochemistry and next generation sequencing (DNA-seq) were used to detect the related protein and gene variation. Patients were followed up.Results:Case 1 was male, 34 years old; and case 2 was female, 28 years old. Both had tumor recurrence in the original site. There were multiple nodules at right calf and ankle, involving superficial subcutaneous tissue and deep interfascicular muscles; some nodules were borderless and painful. Microscopically, the tumor was nodular with fibrous pseudocapsule, some had indistinct borders and diffuse infiltration to the surrounding adipose tissue. The tumor cells were round to ovoid with inconspicuous nucleoli, partly surrounding small irregularly dilated thin-walled blood vessels. The recurrent tumors showed epithelioid morphology in some of the tumor cells, with eosinophilic cytoplasm, some apparent nucleoli, mild to moderate nuclear atypia, and brisk mitotic figures. Focally, perimuscular cell differentiation was noted. The small lesion showed intravascular tumor thrombus. NGS revealed BRAF V600E mutation in case 1, and BRAF V600E mutation combined with PDGFRB gene amplification in case 2.Conclusions:Glomangiomatosis with uncertain malignant potential is a rare variant of glomus tumor. It has a unique growth pattern morphologically, BRAF V600E mutation, and invasive biological behavior.

BACKGROUND:Bone marrow mesenchymal stem cells in patients with osteoporosis show significant senescence and decreased activity and osteogenic differentiation.miR-212-3p inhibits osteogenic differentiation of human bone marrow mesenchymal stem cells.However,its regulation of senescence of bone marrow mesenchymal stem cells and its mechanism remain unclear.OBJECTIVE:To investigate the effect of miR-212-3p on senescence of bone marrow mesenchymal stem cells by targeting mitogen-activated protein kinase 3 (MAPK3) and its mechanism.METHODS:Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro,and the third generation was collected for the following experiments:(1) Cultured in two groups:The control group was added with complete culture medium,and the model group was added with complete culture medium containing H2O2.After 72 hours of culture,β-galactosidase activity,miR-212-3p and MAPK3 mRNA expression,as well as MAPK3,p16,and p21 protein expression were detected.(2) Cultured in three groups:control group,inhibitor control group,and miR-212-3p inhibitor group.After transfection for 24 hours,miR-212-3p,mRNA and protein expression of MAPK3 were detected.(3) Dual luciferase reporter gene combined with qRT-PCR and western blot assay were used to verify the targeting regulation of miR-212-3p and MAPK3.(4) Cultured in different groups:control inhibitor group,miR-212-3p inhibitor group,miR-212-3p inhibitor+interference control group,and miR-212-3p inhibitor+MAPK3 interference group.After transfection for 24 hours,MAPK protein and mRNA expression levels in cells were detected.They were divided into control group,H2O2 group,H2O2+control inhibitor group,H2O2+miR-212-3p inhibitor group,H2O2+miR-212-3p inhibitor+interference control group,and H2O2+miR-212-3p inhibitor+MAPK3 interference group.Cells were transfected for 24 hours and then cultured with H2O2 for 72 hours.Aging-related β-galactosidase activity and p16 and p21 protein expression were detected.RESULTS AND CONCLUSION:(1) Compared with the control group,β-galactosidase activity,miR-212-3p mRNA expression and p16,p21 protein expression were increased in the model group (P<0.05),while MAPK3 mRNA and protein expression levels were decreased (P<0.05).(2) Compared with the control group,the mRNA expression of miR-212-3p was decreased (P<0.05),and the mRNA and protein expression levels of MAPK3 were increased (P<0.05) in miR-212-3p inhibitor group.(3) Double luciferase reporter gene experiment confirmed that MAPK3 was the downstream target gene of miR-212-3p.(4) Compared with the control inhibitor group,the mRNA and protein expression levels of MAPK3 were increased in miR-212-3p inhibitor group (P<0.05).Compared with the miR-212-3p inhibitor group,the mRNA and protein expression levels of MAPK3 in the miR-212-3p inhibitor+MAPK3 interference group were decreased (P<0.05).Compared with H2O2+control inhibitor group,β-galactosidase activity in H2O2+miR-212-3p inhibitor group was decreased (P<0.05).Compared with H2O2+miR-212-3p inhibitor group,β-galactosidase activity in H2O2+miR-212-3p inhibitor+MAPK3 interference group was higher than that in H2O2+miR-212-3p inhibitor group (P<0.05).Compared with the H2O2+control inhibitor group,the protein expression levels of p16 and p21 in the H2O2+miR-212-3p inhibitor group were decreased (P<0.05).Compared with H2O2+miR-212-3p inhibitor group,the protein expression levels of p16 and p21 in H2O2+miR-212-3p inhibitor+MAPK3 interference group were increased (P<0.05).(5) To conclude,downregulation of miR-212-3p inhibits the senescence of rat bone marrow mesenchymal stem cells,and its mechanism of action may be achieved by targeting up-regulation of MAPK3 expression.

Objective:To investigate the clinicopathological features, genetic characteristics, and differential diagnosis of glomangiomatosis with uncertain malignant potential.Methods:Two cases of glomangiomatosis with uncertain malignant potential were collected at Henan Provincial People′s Hospital from 2013 and 2023. Immunohistochemistry and next generation sequencing (DNA-seq) were used to detect the related protein and gene variation. Patients were followed up.Results:Case 1 was male, 34 years old; and case 2 was female, 28 years old. Both had tumor recurrence in the original site. There were multiple nodules at right calf and ankle, involving superficial subcutaneous tissue and deep interfascicular muscles; some nodules were borderless and painful. Microscopically, the tumor was nodular with fibrous pseudocapsule, some had indistinct borders and diffuse infiltration to the surrounding adipose tissue. The tumor cells were round to ovoid with inconspicuous nucleoli, partly surrounding small irregularly dilated thin-walled blood vessels. The recurrent tumors showed epithelioid morphology in some of the tumor cells, with eosinophilic cytoplasm, some apparent nucleoli, mild to moderate nuclear atypia, and brisk mitotic figures. Focally, perimuscular cell differentiation was noted. The small lesion showed intravascular tumor thrombus. NGS revealed BRAF V600E mutation in case 1, and BRAF V600E mutation combined with PDGFRB gene amplification in case 2.Conclusions:Glomangiomatosis with uncertain malignant potential is a rare variant of glomus tumor. It has a unique growth pattern morphologically, BRAF V600E mutation, and invasive biological behavior.

BACKGROUND:Bone marrow mesenchymal stem cells in patients with osteoporosis show significant senescence and decreased activity and osteogenic differentiation.miR-212-3p inhibits osteogenic differentiation of human bone marrow mesenchymal stem cells.However,its regulation of senescence of bone marrow mesenchymal stem cells and its mechanism remain unclear.OBJECTIVE:To investigate the effect of miR-212-3p on senescence of bone marrow mesenchymal stem cells by targeting mitogen-activated protein kinase 3 (MAPK3) and its mechanism.METHODS:Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro,and the third generation was collected for the following experiments:(1) Cultured in two groups:The control group was added with complete culture medium,and the model group was added with complete culture medium containing H2O2.After 72 hours of culture,β-galactosidase activity,miR-212-3p and MAPK3 mRNA expression,as well as MAPK3,p16,and p21 protein expression were detected.(2) Cultured in three groups:control group,inhibitor control group,and miR-212-3p inhibitor group.After transfection for 24 hours,miR-212-3p,mRNA and protein expression of MAPK3 were detected.(3) Dual luciferase reporter gene combined with qRT-PCR and western blot assay were used to verify the targeting regulation of miR-212-3p and MAPK3.(4) Cultured in different groups:control inhibitor group,miR-212-3p inhibitor group,miR-212-3p inhibitor+interference control group,and miR-212-3p inhibitor+MAPK3 interference group.After transfection for 24 hours,MAPK protein and mRNA expression levels in cells were detected.They were divided into control group,H2O2 group,H2O2+control inhibitor group,H2O2+miR-212-3p inhibitor group,H2O2+miR-212-3p inhibitor+interference control group,and H2O2+miR-212-3p inhibitor+MAPK3 interference group.Cells were transfected for 24 hours and then cultured with H2O2 for 72 hours.Aging-related β-galactosidase activity and p16 and p21 protein expression were detected.RESULTS AND CONCLUSION:(1) Compared with the control group,β-galactosidase activity,miR-212-3p mRNA expression and p16,p21 protein expression were increased in the model group (P<0.05),while MAPK3 mRNA and protein expression levels were decreased (P<0.05).(2) Compared with the control group,the mRNA expression of miR-212-3p was decreased (P<0.05),and the mRNA and protein expression levels of MAPK3 were increased (P<0.05) in miR-212-3p inhibitor group.(3) Double luciferase reporter gene experiment confirmed that MAPK3 was the downstream target gene of miR-212-3p.(4) Compared with the control inhibitor group,the mRNA and protein expression levels of MAPK3 were increased in miR-212-3p inhibitor group (P<0.05).Compared with the miR-212-3p inhibitor group,the mRNA and protein expression levels of MAPK3 in the miR-212-3p inhibitor+MAPK3 interference group were decreased (P<0.05).Compared with H2O2+control inhibitor group,β-galactosidase activity in H2O2+miR-212-3p inhibitor group was decreased (P<0.05).Compared with H2O2+miR-212-3p inhibitor group,β-galactosidase activity in H2O2+miR-212-3p inhibitor+MAPK3 interference group was higher than that in H2O2+miR-212-3p inhibitor group (P<0.05).Compared with the H2O2+control inhibitor group,the protein expression levels of p16 and p21 in the H2O2+miR-212-3p inhibitor group were decreased (P<0.05).Compared with H2O2+miR-212-3p inhibitor group,the protein expression levels of p16 and p21 in H2O2+miR-212-3p inhibitor+MAPK3 interference group were increased (P<0.05).(5) To conclude,downregulation of miR-212-3p inhibits the senescence of rat bone marrow mesenchymal stem cells,and its mechanism of action may be achieved by targeting up-regulation of MAPK3 expression.

Objective To propose strategies for developing clinical predictive models,aiming to assist researchers in conducting standardized clinical prediction model studies.Methods Literature review was conducted to summarize the operational steps and content for developing clinical predictive models.Then,a methodological framework was summarized and refined through expert consultation.Results The 11-step methodological framework for developing clinical predictive models was obtained by synthesizing the experience of 456 clinical predictive modeling studies and expert consultation,and the details were analyzed and elaborated.Conclusions This study presents methodological strategies and recommendations for the development of clinical predictive models,intended to serve as a guide for researchers.

Objective To evaluate the clinical application value of portal vein implantation pump for chemotherapy in patients with intrahepatic cholangiocarcinoma (ICC) after radical surgery.Methods The clinical data of 97 patients with ICC who underwent radical surgery in Henan People's Hospital from June 2012 to June 2016 were retrospectively analyzed.Results Among the 97 patients,14 patients received portal venous pump chemotherapy (portal group),33 patients received peripheral venous chemotherapy (peripheral group),and 50 patients did not receive postoperative chemotherapy (control group).There were no statistically significant differences in gender and age between the three groups.The results of survival analysis indicated that the disease-free survival (DFS) period and overall survival (OS) time in the portal group and the peripheral group were significantly better than that in control group (both P ＜ 0.05).In addition,despite the lack of statistical significance (P ＞ 0.05),for the control of intrahepatic metastasis,portal vein pump chemotherapy was better than that of systemic chemotherapy via peripheral vein,and almost all side effects of chemotherapy in the portal group were lower than those in the peripheral group.Conclusion Portal vein pump chemotherapy can improve the prognosis of intrahepatic bile duct patients,especially for the control of intrahepatic metastasis,and can reduce systemic side effects of chemotherapy.

Bone marrow mesenchymal stem cells (BMSCs) have a strong self-renewal ability and multi-directional differentiation potential, so they are ideal cells for cell transplantation, and are widely used in the treatment of diabetes, osteoarthritis and other diseases. BMSCs could increase insulin and C peptide levels in patients by a variety of mechanisms, and a good blood glucose control could be achieved without exogenous insulin therapy. Recently, a number of clinical studies confirmed the safety and effectiveness of BMSCs transplantation in the treatment of diabetes. BMSCs transplantation has also achieved good results for treating complications of diabetes, such as diabetic nephropathy, diabetic peripheral vascular disease, diabetic cardiomyopathy, diabetic bone disease and others. However, due to the lack of standardized transplantation regimens, BMSCs transplantation has been limited in the clinical application. This review summarized the status and research progress of BMSCs transplantation in the treatment of diabetes mellitus and its complications.

Objective:To explore Toll-like receptor 2 (TLR2) and TLR4 polymorphism in Han people from Hunan region and its association with coronary atherosclerotic heart disease.Methods:Sanger sequence and statistical analysis were performed to identify the polymorphism of TLR2 and TLR4 genes in 347 unrelated Hunan Han subjects,including 180 healthy people (control group) and 167 patients with coronary atherosclerotic heart disease (coronary atherosclerotic heart disease group).Results:There was no significant difference in the genotype frequency and allelic frequency for TLR2 SNP2258G＞A and TLR4 SNP896A＞G between the 2 groups (P＞0.05),while there was significant difference in the TLR4 SNP1196C＞T between the 2 groups (P＜0.05).Conclusion:TLR4 SNP 1196C ＞T polymorphism is associated with coronary atherosclerotic heart disease in Chinese Han populationin in Hunan region.

Objective To analyze the genetic characteristics of measles virus in Sichuan province in 2015.Methods Measles virus was isolated from throat swab specimens collected from suspected measles cases.Nucleotide sequences coding for C terminus of nucleoprotein were amplified by RT-PCR and sequenced for phylogenetic analysis.Results 398 measles virus isolates were obtained from 971 throat swab specimens.Phylogenetic analysis showed that 397 belonged to H1 genotype,the sequences homologies were 96.2％-98.0％ compared with H1 genotype reference strain;and all those isolates separated into 2 clusters in phylogenetic tree with the average nucleotide divergence of 2.0％.1 isolate belonged to A genotype,and shared 98.0％ nucleotide homology compared with A genotype reference strain.Conclusions H1 genotype virus remained predominant circulating in Sichuan Province.There was no much genetic variation in N gene.But a minor nucleotide divergence existed between different clusters,leading to 2 transmission chains.