Objective: To analyze the relationship of gross motor skills and perceptual motor abilities with physical activity levels in preschool children in a Beijing kindergarten, so as to provide a reference for promoting the development of motor competence. Methods: From September 2018 to March 2021, preschoolers aged 4-5 years were selected using convenience sampling method from an urban kindergarten in Beijing. The Test of Gross Motor Development-Third Version(TGMD-3) was used to assess basic preschoolers s gross motor skills ( n =152). The Pictorial Scale of Perceived Movement Skill Competence(PMSC) was used to evaluate perceptual motor skills ( n =151). Accelerometers (Actigraph GT3X) were used to record physical activity levels ( n =52). Data were analyzed using one way analysis of variance (ANOVA) and Pearson correlation coefficients. Results: The mean scores for gross motor skills and perceptual motor abilities were (38.76±13.48) and (35.49±6.50), respectively. The moderate to vigorous physical activity(MVPA) level was(52.60±27.44) minutes per day. No statistically significant correlations were found between gross motor skills, perceptual motor abilities MVPA among boys, girls or the overall group ( r =-0.20 to 0.25, all P >0.05). However, Boys locomotor skills, overall children s locomotor skills, and boys gross motor skills were all positively correlated with MVPA( r =0.34-0.45, all P <0.05). Conclusion There is a correlation between locomotor skills and physical activity levels in 4 to 5-year-old children.

Objective:To verify and compare the correlation and diagnostic efficacy of international bowel ultrasound segmental activity score（IBUS-SAS），bowel ultrasound score（BUSS），simple ultrasound score for Crohn's disease（SUS-CD），and simple ultrasound score（Simple-US）with endoscopic disease activity in Crohn's disease（CD）patients. To provide external validation of the diagnostic efficacy of intestinal ultrasound（IUS）score and theoretical basis for clinical selection of optimal IUS score.Methods:A total of 160 patients with clinical diagnosis of CD combined with IUS and intestinal endoscopy were retrospectively analyzed in the Second Affiliated Hospital of Zhejiang University School of Medicine from January 2022 to August 2024. IUS parameters were measured and scored with IBUS-SAS，SUS-CD，BUSS and Simple-US scores. Endoscopic SES-CD was used to evaluate intestinal disease activity in patients without history of intestinal resection，and Rutgeerts score was used to evaluate intestinal disease activity in patients with history of intestinal resection. Endoscopic remission in patients with CD was defined as SES-CD < 3 or Rutgeerts score i0 and i1，mild endoscopic disease activity was defined as 7 > SES-CD≥3 or Rutgeerts score = 2，moderate endoscopic disease activity was defined as 15 > SES-CD≥7 or Rutgeerts score i3，severe endoscopic disease activity was defined as SES-CD≥15 or Rutgeerts i4. The correlation and diagnostic efficacy of IBUS-SAS，SUS-CD，BUSS and Simple-US scores with endoscopic disease activity in patients with CD were compared and analyzed.Results:IUS scores including IBUS-SAS，SUS-CD，BUSS and Simple-US were significantly correlated with endoscopic intestinal disease activity，SES-CD and Rutgeerts scores in CD patients（ r s = 0.706，0.492，0.502，0.526；0.825，0.581，0.584，0.603；0.541，0.434，0.437，0.467；all P＜0.05）. Among them，IBUS-SAS showed better correlation than the other three IUS scores. ROC curve showed that IBUS-SAS，SUS-CD，BUSS，and Simple-US had high predictive values for endoscopic disease activity and endoscopic disease moderate-severe activity in patients with CD（AUC = 0.886，0.748，0.730，0.756；all P＜0.05）. The diagnostic efficacy of IBUS-SAS on the presence of endoscopic disease activity in patients with CD was significantly higher than those of the other three IUS scores . Conclusions:IBUS-SAS，SUS-CD，BUSS and Simple-US are significantly correlated with the endoscopic results of on intestinal disease activity of CD，and have high predictive values for intestinal disease activity status，among which IBUS-SAS is superior to the other three IUS scores. It is recommended that IBUS-SAS be used first to evaluate intestinal disease activity in patients with CD.

Objective To investigate the effects of TEK receptor tyrosine kinase(Tie2)L914F mutation on the biological behavior of vascular endothelial cells and the changes of related signaling pathways in patients with venous malformations.Methods Gene sequen-cing was used to detect Tie2-L914F mutations in venous malformations.HE staining and immunohistochemical staining were used to de-tect the expression of platelet derived growth factor subunit B gene(PDGFB)and α-smooth muscle actin(α-SMA)in venous malfor-mations caused by the mutation.Lentivirus overexpressing Tie2-wild type(Tie2-WT),Tie2-L914F and Tie2-GFP(green fluorescent protein,GFP)infected human umbilical vein endothelial cells(HUVECs).Real-time fluorescence quantitative PCR was used to detect the expression level of Tie2 in endothelial cells expressing exogenous Tie2 and Flag-tagged protein Tie2 protein was detected by western blotting to verify transfection efficiency.The proliferation,apoptosis,migration and tube-forming ability of the cells were determined by CCK-8,flow cytometry,Transwell migration assay and Matrigel matrix gel tube-forming assay.Western blotting was used to detect the expression levels of Protein Kinase B(PKB/AKT),FOXO1 and their phosphorylation,and the expression level of PDGFB was detec-ted by ELISA.Results The number of patients with venous malformations with L914F mutations was about 33.3％.HE and immuno-histochemical staining showed that the expressions of PDGFB and α-SMA were significantly down-regulated in venous malformation tis-sues with Tie2-L914F mutation,and were positively correlated with the decrease in cell coverage of the tube wall.Compared with Tie2-WT endothelial cells,the apoptosis number of Tie2-L914F endothelial cells was significantly reduced,while the proliferation and mi-gration ability was significantly increased,and the tube-forming ability was significantly decreased.Western blotting and ELISA showed that the phosphorylation levels of AKT and FOXO1 downstream of Tie2 signaling pathway in endothelial cells expressing Tie2-L914F were significantly increased,and the expression level of PDGFB was significantly decreased.Conclusion In venous malformations,Tie2-L914F mutation may downregulate the expression of PDGFB through AKT signaling pathway,which affects the biological behavior of vascular endothelial cells.

Objective:To verify and compare the correlation and diagnostic efficacy of international bowel ultrasound segmental activity score（IBUS-SAS），bowel ultrasound score（BUSS），simple ultrasound score for Crohn's disease（SUS-CD），and simple ultrasound score（Simple-US）with endoscopic disease activity in Crohn's disease（CD）patients. To provide external validation of the diagnostic efficacy of intestinal ultrasound（IUS）score and theoretical basis for clinical selection of optimal IUS score.Methods:A total of 160 patients with clinical diagnosis of CD combined with IUS and intestinal endoscopy were retrospectively analyzed in the Second Affiliated Hospital of Zhejiang University School of Medicine from January 2022 to August 2024. IUS parameters were measured and scored with IBUS-SAS，SUS-CD，BUSS and Simple-US scores. Endoscopic SES-CD was used to evaluate intestinal disease activity in patients without history of intestinal resection，and Rutgeerts score was used to evaluate intestinal disease activity in patients with history of intestinal resection. Endoscopic remission in patients with CD was defined as SES-CD < 3 or Rutgeerts score i0 and i1，mild endoscopic disease activity was defined as 7 > SES-CD≥3 or Rutgeerts score = 2，moderate endoscopic disease activity was defined as 15 > SES-CD≥7 or Rutgeerts score i3，severe endoscopic disease activity was defined as SES-CD≥15 or Rutgeerts i4. The correlation and diagnostic efficacy of IBUS-SAS，SUS-CD，BUSS and Simple-US scores with endoscopic disease activity in patients with CD were compared and analyzed.Results:IUS scores including IBUS-SAS，SUS-CD，BUSS and Simple-US were significantly correlated with endoscopic intestinal disease activity，SES-CD and Rutgeerts scores in CD patients（ r s = 0.706，0.492，0.502，0.526；0.825，0.581，0.584，0.603；0.541，0.434，0.437，0.467；all P＜0.05）. Among them，IBUS-SAS showed better correlation than the other three IUS scores. ROC curve showed that IBUS-SAS，SUS-CD，BUSS，and Simple-US had high predictive values for endoscopic disease activity and endoscopic disease moderate-severe activity in patients with CD（AUC = 0.886，0.748，0.730，0.756；all P＜0.05）. The diagnostic efficacy of IBUS-SAS on the presence of endoscopic disease activity in patients with CD was significantly higher than those of the other three IUS scores . Conclusions:IBUS-SAS，SUS-CD，BUSS and Simple-US are significantly correlated with the endoscopic results of on intestinal disease activity of CD，and have high predictive values for intestinal disease activity status，among which IBUS-SAS is superior to the other three IUS scores. It is recommended that IBUS-SAS be used first to evaluate intestinal disease activity in patients with CD.

Objective To explore the clinical value of apolipoprotein C1(APOC1)and Klotho expression levels in predicting the prognosis of patients with renal cancer after laparoscopic partial nephrectomy.Methods Eighty patients diagnosed as renal cell carcinoma and underwent laparoscopic partial nephrectomy were collected as the study subjects.According to the prognosis,patients were separated into the good prognosis group(61 cases)and the poor prognosis group(19 cases).Seventy-eight healthy individuals underwent physical examination were collected as the control group.The general clinical data of the subjects were collected,and serum levels of APOC1 and Klotho were tested and analyzed in three groups.Pearson method was applied to analyze the correlation between serum APOC1 and Klotho levels in patients.Cox regression was applied to analyze factors affecting the prognosis of patients.Receiver operating characteristic(ROC)curve was applied to analyze the predictive efficacy of APOC1 and Klotho levels on the prognosis of patients.Results There were significant differences in clinical stage and pathological grade between the poor prognosis group and the good prognosis group(P＜0.05).Compared with the control group,the serum APOC1 levels were significantly increased in the good prognosis group and the poor prognosis group(P＜0.05),while the Klotho expression levels were obviously reduced(P＜0.05).And the serum APOC1 level in the control group,the poor prognosis group and the good prognosis group was increased successively(P＜0.05),while the serum Klotho level was obviously decreased successively(P＜0.05).The serum APOC1 expression level of patients was negatively correlated with Klotho level(r=-0.577,P＜0.001).The Cox regression results showed that decreased expression level of Klotho and elevated expression level of APOC1,pathologic grading 2 and clinical staging stage Ⅱ were all independent risk factors for poor prognosis(P＜0.05).The area under the curve(AUC)of serum APOC1 and Klotho levels,and their combined application in predicting poor postoperative prognosis in renal cancer patients was 0.863,0.850 and 0.953,respectively,and the clinical value of combination of the two in predicting the prognosis of patients was superior to that of APOC1 and Klotho alone.Conclusion Patients with poor prognosis after retroperitoneal laparoscopic partial nephrectomy have a obvious increase in serum levels of APOC1 and a obvious decrease in serum level of Klotho.The combination of the two has high clinical significance in predicting the prognosis of patients with retroperitoneal laparoscopic partial nephrectomy.

Objective To investigate the effects of TEK receptor tyrosine kinase(Tie2)L914F mutation on the biological behavior of vascular endothelial cells and the changes of related signaling pathways in patients with venous malformations.Methods Gene sequen-cing was used to detect Tie2-L914F mutations in venous malformations.HE staining and immunohistochemical staining were used to de-tect the expression of platelet derived growth factor subunit B gene(PDGFB)and α-smooth muscle actin(α-SMA)in venous malfor-mations caused by the mutation.Lentivirus overexpressing Tie2-wild type(Tie2-WT),Tie2-L914F and Tie2-GFP(green fluorescent protein,GFP)infected human umbilical vein endothelial cells(HUVECs).Real-time fluorescence quantitative PCR was used to detect the expression level of Tie2 in endothelial cells expressing exogenous Tie2 and Flag-tagged protein Tie2 protein was detected by western blotting to verify transfection efficiency.The proliferation,apoptosis,migration and tube-forming ability of the cells were determined by CCK-8,flow cytometry,Transwell migration assay and Matrigel matrix gel tube-forming assay.Western blotting was used to detect the expression levels of Protein Kinase B(PKB/AKT),FOXO1 and their phosphorylation,and the expression level of PDGFB was detec-ted by ELISA.Results The number of patients with venous malformations with L914F mutations was about 33.3％.HE and immuno-histochemical staining showed that the expressions of PDGFB and α-SMA were significantly down-regulated in venous malformation tis-sues with Tie2-L914F mutation,and were positively correlated with the decrease in cell coverage of the tube wall.Compared with Tie2-WT endothelial cells,the apoptosis number of Tie2-L914F endothelial cells was significantly reduced,while the proliferation and mi-gration ability was significantly increased,and the tube-forming ability was significantly decreased.Western blotting and ELISA showed that the phosphorylation levels of AKT and FOXO1 downstream of Tie2 signaling pathway in endothelial cells expressing Tie2-L914F were significantly increased,and the expression level of PDGFB was significantly decreased.Conclusion In venous malformations,Tie2-L914F mutation may downregulate the expression of PDGFB through AKT signaling pathway,which affects the biological behavior of vascular endothelial cells.

Objective To explore the clinical value of apolipoprotein C1(APOC1)and Klotho expression levels in predicting the prognosis of patients with renal cancer after laparoscopic partial nephrectomy.Methods Eighty patients diagnosed as renal cell carcinoma and underwent laparoscopic partial nephrectomy were collected as the study subjects.According to the prognosis,patients were separated into the good prognosis group(61 cases)and the poor prognosis group(19 cases).Seventy-eight healthy individuals underwent physical examination were collected as the control group.The general clinical data of the subjects were collected,and serum levels of APOC1 and Klotho were tested and analyzed in three groups.Pearson method was applied to analyze the correlation between serum APOC1 and Klotho levels in patients.Cox regression was applied to analyze factors affecting the prognosis of patients.Receiver operating characteristic(ROC)curve was applied to analyze the predictive efficacy of APOC1 and Klotho levels on the prognosis of patients.Results There were significant differences in clinical stage and pathological grade between the poor prognosis group and the good prognosis group(P＜0.05).Compared with the control group,the serum APOC1 levels were significantly increased in the good prognosis group and the poor prognosis group(P＜0.05),while the Klotho expression levels were obviously reduced(P＜0.05).And the serum APOC1 level in the control group,the poor prognosis group and the good prognosis group was increased successively(P＜0.05),while the serum Klotho level was obviously decreased successively(P＜0.05).The serum APOC1 expression level of patients was negatively correlated with Klotho level(r=-0.577,P＜0.001).The Cox regression results showed that decreased expression level of Klotho and elevated expression level of APOC1,pathologic grading 2 and clinical staging stage Ⅱ were all independent risk factors for poor prognosis(P＜0.05).The area under the curve(AUC)of serum APOC1 and Klotho levels,and their combined application in predicting poor postoperative prognosis in renal cancer patients was 0.863,0.850 and 0.953,respectively,and the clinical value of combination of the two in predicting the prognosis of patients was superior to that of APOC1 and Klotho alone.Conclusion Patients with poor prognosis after retroperitoneal laparoscopic partial nephrectomy have a obvious increase in serum levels of APOC1 and a obvious decrease in serum level of Klotho.The combination of the two has high clinical significance in predicting the prognosis of patients with retroperitoneal laparoscopic partial nephrectomy.

Bone diseases, such as osteoporosis and osteoarthritis, have emerged as pressing public health concerns requiring immediate attention and resolution. Cellular therapy and tissue engineering techniques are among the most promising therapeutic approaches for such conditions. Human induced pluripotent stem cells (hiPSCs) possess remarkable capacity for indefinite self-renewal in vitro and the ability to differentiate into all somatic cell types originating from the three germ layers, thereby making them a promising source of osteoblasts. Consequently, it is crucial to establish a well-delineated system for osteogenic differentiation of hiPSCs in vitro, with the aim to generate osteoblast-like cells that conform to clinical application standards. Numerous research teams have achieved substantial advancements in both the direct osteogenic differentiation of hiPSCs and the indirect pathway via mesenchymal stem cells. In this article, we provide a comprehensive review of these two osteogenic differentiation pathways and their current applications, with the aim of serving as a valuable reference for bone regeneration technologies. Current research efforts have relied on embryoid body formation and monolayer induction methods utilizing biomaterials to develop a system that facilitates in vitro culture and osteogenic differentiation of hiPSCs. However, the existing research is primarily constrained by unclear system components and low efficiency. Therefore, the development of a stepwise and three-dimensional induction system based on stringent regulation by specific compounds is a primary research direction for the future.

BACKGROUND: The addition of growth factiors is commonly applied to improve the osteogenic differentiation of stem cells. However, for human pluripotent stem cells (hPSCs), their complex differentiation processes result in the unknown effect at different stages. In this study, we focused on the widely used bone forming peptide-1 (BFP-1) and investigated the effect and mechanisms of its addition on the osteogenic induction of hPSCs as a function of the supplementation period. METHODS: Monolayer-cultured hPSCs were cultured in osteogenic induction medium for 28 days, and the effect of BFP-1 peptide addition at varying weeks was examined. After differentiation for varying days (0, 7, 14, 21 and 28), the differentiation efficiency was determined by RT–PCR, flow cytometry, immunofluorescence, and alizarin red staining assays. Moreover, the expression of marker genes related to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7. RESULTS: Peptide treatment during the first week promoted the generation of mesoderm cells and mesenchymal-like cells from hiPSCs. Then, the upregulated expression of osteogenesis marker genes/proteins was detected in both hESCs and hiPSCs during subsequent inductions with BFP-1 peptide treatment. Fortunately, further experimental design confirmed that treating the BFP-1 peptide during 7–21 days showed even better performance for hESCs but was ineffective for hiPSCs. CONCLUSION The differentiation efficiency of cells could be improved by determining the optimal treatment period.Our study has great value in maximizing the differentiation of hPSCs by adding osteogenesis peptides based on the revealed mechanisms and promoting the application of hPSCs in bone tissue regeneration.

0bjective:To observe The effects of 12-week Tai Chi Cardiac Rehabilitation Programme(TCCRP)and conven-tional exercise rehabilitation program(CERP)on inflammatory factors and adhesion molecules in patients with coronary heart disease(CHD).Method:Forty-six patients with CHD were jointly recruited from Anzhen Community Service Center at Bei-jing Chaoyang District and Wanjie Rehabilitation Hospital at Shandong Zibo from Oct.2020 to Nov.2021,and were randomly divided into the TCCRP group(n=23)and the CERP group as the control group(n=23)ac-cording to a 1∶1 ratio.Thirty-four patients completed the whole program,14 patients in the TCCRP group and 20 patients in the CERP group.All program were 60 min per time,3 times a week for a total of 12 weeks,including 4 weeks of in-hospital rehabilitation and 8 weeks of remote on-line home-based rehabilita-tion.Inflammatory factors and adhesion molecules were detected before and after the intervention.Result:After 12 weeks of intervention,C-reactive protein(CRP),interleukin 6(IL-6),vascular cell adhe-sion molecule(VCAM-1),and intercell adhesion molecule(ICAM-1)all decreased significantly(P＜0.01).The anti-inflammatory factor interleukin 10(IL-10)increased significantly in the TCCRP group(P＜0.0l).There was no significant difference between the two groups before the intervention(P＞0.05),and the VCAM-1 in the TCCRP group was significantly lower than the CERP group after the intervention(P＜0.05).Conclusion:In patients with coronary heart disease,the 12-week TCCRP and CERP intervention can effectively reduce the levels of pro-inflammatory factors CRP,IL-6 and adhesion molecules ICAM-1 and VCAM-1.The TC-CRP can effectively improve the level of anti-inflammatory factor IL-10 and reduce the expression of adhesion molecule VCAM-1 compared with CERP,which indicate a more positive effect on regulating the balance between pro-inflammatory and anti-inflammatory factors and improving the anti-inflammatory and anti-adhesion ability.