Objective: To analyze the impact of parental sports concept and behavior on physical activity in preschool children, so as to provide a foundation for future guidance on fostering childrens physical activity within the family context. Methods: From November to December 2020, a clustered convenience sampling method was employed to conduct surveys, and a total of 283 children were selectal from one kindergarten each in Beijing, Shenyang, and Xian. Participating children wore ActiGraph GT9X accelerometers continuously for one week to collect data on different intensity levels of physical activity. Physical Activity afterschool Questionnaire for Preschooler (P-PAQ) was utilized to assess parental sports concept and behavior. The gender differences in physical activity level and physical activity compliance rate were analyzed by using ttest, Mann-Whitney U test, and Chisquare test; and the relationship between parental exercise concepts and behaviors and physical activity of preschool children was analyzed using Spearman rank correlation analysis and multiple linear regression analysis. Results: Parental sports concept was significantly positively correlated with average daily moderatetovigorous physical activity (MVPA) and total physical activity (TPA) in children (r=0.12-0.16, P<0.05). Parental sports behavior was significantly positively correlated with childrens average daily TPA (r=0.25, P<0.05). Multiple linear regression revealed that parental sports concept was positively correlated with average daily MVPA and TPA in both boys and girls (B=0.65-0.83), while parental sports behavior only was positively correlated with boys average daily MVPA and TPA (B=0.24-0.25)(P<0.05). Conclusions Parental sports concept and behavior can impact physical activity levels in preschool children, exhibiting gender differences. Future guidance on physical activity in family upbringing should consider both parental sports concept and behavior, and pay attention to the influence of childrens gender.

Objective: To establish the norm of the Physical Activity afterschool Questionnaire for Preschooler(P-PAQ) in urban areas of China, so as to provide a basis for graded guidance from the family perspective and to improve children s physical activity levels. Methods: From October 2020 to January 2021, 6 267 children aged 3-6 years old were recruited from 40 kindergartens in eight cities across six major administrative regions by stratified cluster sampling, and the P-PAQ initially developed by the researchers of this study were completed by the primary caregivers. The questionnaire was administered to collect data relating to the amount of physical activity undertaken by the preschoolers, and the norm was determined by quartiles. Data relating to parental concepts of sports and parental behavior were assessed by calculating mean scores in order to establish the norm. Results: Among preschoolers in urban areas, the M(P 25 ，P 75 ) of total physical activity time (min/day), moderate-to-vigorous physical activity time (min/day), outdoor time (min/day) and screen time (min/day) on school days outside kindergarten and on weekends were 84 (54,120), 22 (8,40), 12 (0,24) and 18 (6,30), and 170 (115,240), 60 (30,95), 90 (35,120) and 30 (20,60), respectively. When the score of parents sports concept and behavior (total score of 40) were≥34, 29-<34, 24-<29, <24, it was defined as four levels about above medium, medium, lower medium and lower, respectively. And for two dimensions，when the score of parental sports concept were ≥19, 17-<19, 15-<17, <15，and the score of parental behaviors were ≥16, 12-<16, 8-<12, <8, it was defined as four levels about upper medium, medium, lower medium and lower, respectively. Conclusion The norm of extracurricular activities among preschool children in Chinese cities has good representativeness and appropriate threshold values, which could provide a valuable reference for early assessment, as well as guidance in relation to out-of-school physical activity behaviors among children aged 3-6 years old.

Objective:To clarify the effect and related factors of antiviral therapy on the change of esophageal varices in patients with hepatitis B virus-related cirrhosis.Methods:Fifty-two cases with hepatitis B virus-related cirrhosis who underwent endoscopy before and after antiviral therapy were selected from prospective cohorts. Patients were divided into three groups: no, mild, and moderate-severe based on the degree of esophageal varices. The changes in the severity of esophageal varices in each group were compared after antiviral therapy. Clinical characteristics (platelet, liver and kidney function, liver stiffness, and virological response) of patients with different regressions were analyzed. Measurement data were analyzed by independent sample t-test, one-way ANOVA, Mann-Whitney U test and Kruskal-Wallis H test, and Chi-Square test was used for count data.Results:All patients received entecavir-based antiviral therapy. The median treatment time was 3.1 (2.5-4.4) years. The proportion of patients without esophageal varices increased from 30.8% to 51.9%, the proportion of mild esophageal varices decreased from 40.4% to 30.8%, and the proportion of patients with moderate-to-severe esophageal varices decreased from 28.8% to 17.3% ( χ2=14.067, P=0.001). A total of 40.4% of patients had esophageal varices regression, and 13.5% had esophageal varices progression. The progression rate was significantly higher in patients with moderate-severe esophageal varices than patients with mild and no esophageal varices ( χ2=28.126, P<0.001), and 60.0% of patients with moderate-severe esophageal varices still remained in moderate-severe state after antiviral treatment. Baseline platelet count and 5-year mean change rates were significantly lower in patients with progressive moderate-to-severe esophageal varices than in those without progression (+3.3% vs. +34.1%, Z=7.00, P=0.027). Conclusion:After effective antiviral treatment, 40.4% of patients with hepatitis B virus-related cirrhosis combined with esophageal varices has obtained esophageal varices regression, but those with moderate to severe esophageal varices still have a considerable risk of progression while receiving mono antiviral treatment only. Thrombocytopenia and without significant improving are the clinical signs of progression risk after receiving antiviral treatment.

Objective:To investigate the related mechanism of miRNA-34a (miR-34a) reverses cisplatin resistance of osteosarcoma through targeted inhibition of high mobility group box 1 (HMGB1).Methods:The MG-63 cisplatin-resistant cell line (MG-63/DDP) was constructed by using dose escalation and intermittent action, and then the successfully constructed MG-63/DDP cells were treated with different concentrations of cisplatin (0, 1, 5, 10, 20, 30 μg/ml), and CCK-8 method was used to detect cell survival rate. The MG-63/DDP cells were transfected respectively and then randomly divided into two groups: the miR-34a overexpression vector group and the miR-34a empty expression vector (miR-34a-NC) group. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression level of miR-34a. Transfected cells were treated with different concentrations of cisplatin (0, 1, 5, 10, 20, 30 μg/ml), and CCK-8 method was used to detect cell survival rate, flow cytometry was used to detect cell apoptosis. The dual luciferase reporter gene experiment was used to verify whether miR-34a regulated the expression of HMGB1, and Western blotting method was used to detect the HMGB1 protein expression level of the transfected cells. MG-63/DDP cells were transfected respectively and then randomly divided into two groups: HMGB1 gene silencing vector (si-HMGB1) group and its negative control vector (si-NC) group. Western blotting method was used to detect HMGB1 protein expression level and CCK-8 method was used to detect cell survival rate.Results:The MG-63/DDP cell line was successfully constructed. The survival rate of MG-63/DDP cells was higher than that of MG-63 cells when cells were treated with different concentrations of cisplatin (all P < 0.01), and half inhibitory concentration ( IC50) value of MG-63/DDP cells and MG-63 cells on cisplatin was 25.80 μg/ml and 0.47 μg/ml, respectively. qRT-PCR results showed that the relative expression level of miR-34a in MG-63/DDP cells was lower than that in MG-63 cells (0.46±0.04 vs. 1.02±0.05, t = 15.14, P < 0.01); compared with miR-34a-NC group, the relative expression of miR-34a in MG-63/DDP cells was increased in miR-34a overexpression vector group (1.67±0.09 vs. 1.00±0.02, t = -12.58, P < 0.01). Cell survival rate of miR-34a overexpression vector group and miR-34a-NC group was decreased with the increase in the concentration of cisplatin; cell survival rate of miR-34a overexpression vector group was lower than that of miR-34a-NC group when cells were treated with different concentrations of 5- 30 μg/ml cisplatin (all P < 0.01). The apoptotic rate of MG-63/DDP cells in miR-34a-NC group and miR-34a overexpression vector group was (25.1±1.7)% and (42.3±2.3)%, respectively when cells were treated with 20 μg/ml cisplatin; and in miR-34a overexpression vector group, MG-63/DDP cells had a higher rate of apoptosis ( P < 0.01). The dual luciferase reporter gene experiment results showed that compared with miR-34a-NC group, miR-34a overexpression vector group could inhibit the luciferase activity of PGL3- wild-type-HMGB1, and the difference was statistically significant ( t = 6.37, P < 0.01), while miR-34a overexpression vector group had no significant inhibitory effect on the luciferase activity of PGL3- mutant-HMGB1 ( t = 0.35, P = 0.74). The relative expression level of HMGB1 protein in miR-34a overexpression vector group was lower than that in miR-34a-NC group (0.43±0.02 vs. 1.00±0.14, t = 6.98, P < 0.01). Compared with si-NC group, the relative expression level and IC50 value of HMGB1 protein in si-HMGB1 group were reduced (all P < 0.05). Conclusion:Overexpression of miR-34a can enhance the chemosensitivity of osteosarcoma cells MG-63/DDP to cisplatin, and its mechanism may be related to the inhibition of HMGB1 expression.

Objective: To investigate the effect and mechanism of AMP-activated protein kinase α (AMPKα) over-expression on proliferation, migration, invasion and epithelial mesenchymal transition (EMT) of bladder cancer T24 cells. Methods: A bladder cancer T24 cells over-expressing AMPKα was established and divided into T24 group, pc-DNA group and pc-AMPKα group according to different plasmid transfection. Western blotting was used to verify the over-expression ofAMPKα and detect the expressions of EMT-related proteins and EMT pathway-related molecules. Hoechst staining was used to detect apoptosis of transfected T24 cells. CCK8 assay was used to detect cell proliferation. Cell scratch test was used to detect cell migration. Transwell assay was used to detect cell invasion. Results: The bladder cancer cell line T24 over-expressingAMPKα was successfully constructed. Compared with the T24 group and the pc-DNA group, the level of E-cadherin in the pc-AMPKα group was significantly increased (P<0.01) while the levels of Vimentin and N-cadherin were significantly decreased (all P<0.01), and the activities of P38 and STAT3 which related to EMT pathway were significantly inhibited (all P<0.01); cell proliferation, migration and invasion were significantly decreased while cell apoptosis was obviously enhanced (all P<0.01). Conclusion: Over-expression of AMPKα can inhibit the activity of EMT pathway-related molecules, which leads to obvious apoptosis, limited proliferation, reduced invasion and migration of bladder cancer T24 cells, and accompanied by the reversal of EMT.

Objective To evaluate the potential value of next-generation sequencing ( NGS) as a diagnostic method for listeria infection in central nervous system ( CNS ) . Methods To identify the potentially pathogenic microorganisms ( PPMs) of the five patients with CNS infection in the Department of Neurology, People's Hospital of Zhengzhou University from June 2017 to November 2017, blood or cerebrospinal fluid ( CSF ) was detected not only using common laboratory tests , including routine , biochemical, cytologic, culture and Gram-staining methods, etc, but also using the BGISEQ-100 sequencing platform to identify the PPMs of CSF .Concomitantly , we collected concurrent clinical data , then performed a comprehensive analysis of their clinical , laboratory , and auxiliary examination data .Results Of the five cases (male :female: 1:4, age ranges: 26 -61 years), the disease mainly occurred in summer.Three patients received immunosuppressants treatment before infection , and three patients had gastrointestinal syndrome in the prodromal period .Infection of CNS led to fever , headache and meningeal irritation sign in all the patients.The medical imaging examinations showed the invasion of meninges , brainstem, and the periventricular gray matter.The cell count of CSF was more than 500 ×106/L.NGS techniques showed listeria genome sequence ranged from 57 to 2611, and the coverage of listeria varied from 0.23％ to 14.00％, and PCR results supported the existence of listeria .Conclusion NGS is beneficial to make the diagnosis of listeria infection in CNS , and can help guide early treatment .

Objective To analyze the clinical and electrophysiological features in a family with spinal muscular atrophy (SMA), and assess the probable causative gene mutations for the family. Methods To identify the nosogenesis of the proband with weakness and atrophy in the double lower proximal limbs, clinical data of his 12 family members were collected, and the proband and his mother were selected for clinical examinations, including laboratory tests, electromyogram (EMG), F-wave, H-reflex, X-ray of the spine and double lower limbs, brain and spinal cord magnetic resonance imaging, etc. Moreover, human whole exome sequencing was performed on blood sample from the proband, then its deleterious effects were assessed according to the Standards and guidelines for the interpretation of sequence variants, a joint consensus recommendation of the American College of Medical Genomics (ACMG) and the Association for Molecular Pathology (AMP). Subsequently, the strong pathogenic mutation was validated by Sanger sequencing. Results Familial investigation showed seven of 12 family members presented with weakness in the double lower proximal limbs. Among them, three had the main manifestation of atrophy in the double lower proximal limbs, one had high arched foot as the main presentation, and the others had weakness in the double lower proximal limbs. EMG studies showed the abnormal results in the anterior horn of the spinal cord. The strong pathogenic mutation in DYNC1H1 gene (exon8, c.2327C＞T, p.P776L) was identified from the proband according to ACMG and AMP guidelines. Sanger sequencing revealed six patients had this variant and it was passed mainly from his maternal grandmother. Conclusions A pathogenic mutation of the DYNC1H1 p.P776L in six Chinese pedigrees which cosegregated with SMA was identified. There existed individual differences in clinical presentations. This finding may have important implications for the study of SMA in Chinese patients.