Objective To study the effects of Genistein - magnetic - nanoparticles at different concentrations on the growth and ex-pression of focal adhesion kinase (FAK) in human hepatocellular carcinoma cell line HcpG2. Methods Activities of HepG2 cells treated by Genistein - magnetic - nanoparticles were examined by MTT assay. The expression of FAK mRNA and protein was respectively detected by immunohistochemistry staining and reversed transcriptase polymerase chain reaction (RT - PCR) . Results Genistein - magnetic -nanoparticles at a concentration of 10 -80mg/L inhibited the proliferation of HepG2 cells, with obvious concentration - dapendent and time - dependent effect relationships (P < 0.05). After treatment with various concentrations of Genistein - magnetic - nanoparticles for 48h, the relative level of FAKmRNA of Genistein - magnetic - nanoparticles and control groups was 1.242 ± 0.031,1.213 ± 0.443,0.834 ± 0.044,0.652 ± 0.039,0.446 ± 0.041, and 1.443 ± 0.053 (F = 21.97 ,P < 0.05), respectively. The expression of FAK protein in the cells was decreased, which showed an obvious a concentration - effect relationship. Conclusion Genistein - magnetic - nanoparticles can reduce the mRNA and protein expressions of FAK and inhibit the proliferation of HepG2 cells.

Objective To study the effects of Genistein-magnetic-nanoparticles at different concentrations on the growth and expression of focal adhesion kinase(FAK) in human hepatocellular carcinoma cell line HepG2.Methods Activities of HepG2 cells treated by Genistein-magnetic-nanoparticles were examined by MTT assay.The expression of FAK mRNA and protein was respectively detected by immunohistochemistry staining and reversed transcriptase polymerase chain reaction(RT-PCR).Results Genistein-magnetic-nanoparticles at a concentration of 10-80mg/L inhibited the proliferation of HepG2 cells,with obvious concentration-dapendent and time-dependent effect relationships(P

Objective To investigate the effects of capsaicin on the invasion ability of human large cell carcinoma NCI-H460 and the expressions of E-cadherin and Snail;To discuss the possible mechanisms of anti-non-small cell lung cancer.Methods NCI-H460 cells were cultured in vitro and treated with capsaicin at various concentrations, and no capsacin-treated group was set as control group. Effects of capsaicin on NCI-H460 apoptosis, its invasion ability, and the changes in protein expressions of E-cadherin and Snail were evaluated by Hoechst33342 nuclear staining assay, Transwell chamber invasion assay, and Western blot respectively. Results Compared with the control group, Hoechst33342 nuclear staining assay showed that capsaicin could induce NCI-H460 cell apoptosis (P<0.05);Transwell invasion in vitro results showed that capsaicin could significantly inhibit invasion of penetrating cells (P<0.05);Western blot analysis showed that E-cadherin expression level was significantly elevated and snail expression level significantly decreased (P<0.05).Conclusion Capsaicin can induce NCI-H460 cell apoptosis. Decrease the Snail expression and stimulate E-cadherin expression so as to inhibit the invasion ability of NCI-H460, which may be one of its mechanisms of anti-non-small cell lung cancer.

Objective To investigate the pathogenic characteristics of Vibrio cholera strains isola-ted from Hubei province in 2012 , and to identify the source of infection by analyzing their genetic correla-tions.Methods The biochemical identification , toxin gene detection and drug susceptibility test were car-ried out to analyze a total of 35 Vibrio cholera strains isolated from three epidemic areas .Comparison of ge-nomic DNA fingerprints and cluster analysis among isolates of Vibrio cholera was conducted by using pulsed-field gel electrophoresis ( PFGE ) .Results All of the 35 strains were Vibrio cholera O139 , of which 71.42%were toxic strains.The drug resistance rates of Vibrio cholera strains to tetracycline, cotrimoxazole and rifampincin were 57.14%, 88.57%and 80.00%, respectively.Analysis of genomic DNA fingerprints of the isolates showed highly similar with similarity values ranging from 80%-100%.Most of the strains iso-lated from the same epidemic area fell into the same one cluster with 100% homology in genome Only a strain isolated from turtle in Jingzhou area was belong to a different cluster .Conclusion The Vibrio cholera O139 strains were the dominant strains causing the outbreaks of cholera in Hubei province in 2012 .Most of them were toxigenic strains .A large majority of the strains had developed resistance to tetracycline , cotri-moxazole and rifampincin , but all strains showed high susceptibility to ceftriaxone and imipenem .Vibrio cholera strains isolated from the same epidemic area were mainly belonged to the same one cluster , sugges-ting the same source of infection .However, the strains varied among different epidemic area .Follow-up in-vestigations of three outbreaks of cholera in this study were all associated with food infection .Therefore , more attention should be paid to food sanitation and safety measurement .Although a non-toxigenic strain iso-lated from turtle was not associated with the epidemic of cholera , surveillance for seafood and aquatic prod-ucts would still be necessary .

Objective To investigate the change of serum cystatin C (CC) level and its clinical significance in elderly patients with obstructive sleep apnea hypopnea syndrome (OSAHS),and to evaluate the diagnostic value of serum CC for renal impairment in elderly OSAHS patients.Methods Elderly snoring subjects (age ≥60 years) undergoing sleep apnea monitoring in our sleep laboratory were recruited.Patients were divided into simple snoring group,mild OSAHS group,moderate OSAHS group and severe OSAHS group according to the diagnostic criteria of American academy of sleep medicine.Anthropometric characteristics,lipid profiles,blood glucose,creatinine,estimated glomerular filtration rate (eGFR) and serum CC were compared between groups after age,gender and basic diseases were adjusted.The independent risk factors associated with serum CC were analyzed by Spearman's rank correlation and stepwise linear regression.Results Totally 89 elderly patients [70males and 29 females,aged (68.4±6.3) years in average] were collected.21 subjects were grouped as simple snoring,14 subjects as mild OSAHS,21 subjects as moderate OSAHS and 33 subjects as severe OSAHS.There were no differences in age,gender,basic diseases (hypertension and diabetes),lipid profiles,creatinine and eGFR among groups.The levels of body mass index,systolic blood pressure,diastolic blood pressure and serum CC were higher in severe OSAHS group than in simple snoring group [(28.3±3.9) vs.(24.6±5.9),(136.55±16.14) mm Hgvs.(122.43±16.64) mm Hg,(89.18±11.92) mm Hgvs.(78.10±9.55) mm Hg,(0.95±0.22) mg/L vs.(0.76±0.13)mg/L,respectively,all P＜0.05].Serum CC was significantly correlated with apnea hypopnea index (AHI),oxygen desaturation index,creatinine,eGFR,high sensitive C reactive protein (r=0.397,0.347,0.275,－0.354,0.254 respectively,all P％0.05).Regression analysis showed that AHI was an independent factor for serum CC (β=0.449,P＜ 0.001).Conclusions Serum CC level is increased in elderly patients with severe OSAHS.AHI is an independent factor for the increase of serum CC.Serum CC can be as an index for the early impairment of renal function in patients with OSAHS.

Objective To evaluate the efficacy of eucalyptol-limonene-pinene enteric capsule on stable chronic obstructive pulmonary disease (COPD) in the elderly.Methods A total of 120 patients with stable COPD were randomized into ELP group (n=63) with conventional treatment combined ELP and control group (n=57) with conventional treatment without ELP.Primary outcome was the number of exacerbations per year, and secondly outcomes were lung function parameters and St.George's Respiratory Questionnaire (SGRQ).Results After 1 year of treatment, numbers of exacerbations per patient per year were lower in ELP group than in control group (0.70 times/y vs.1.21 times/y, Z=-3.887, P=0.000).The proportion of exacerbation-free patients were 25.4％(16/63) in ELP group and 5.3％ (3/57) in control group (x2 =9.103, P=0.003).The differences in the forced vital capacity (FVC), forced expiratory volume in one second (FEV1), and FEV1/ FVC ratio (FEV1％) between 5 different time points measurements were statistically significant in both groups (ELP group: F=15.429, 36.389, 33.278, respectively, all P=0.000;control group: F=54.238, 94.213 and 83.774, respectively, all P=0.000).Difference in FEV1/FVC ratio between 5 different time point was statistically significant in control group(F=2.766, P=0.043), but not in ELP group(F=0.861, P=0.451).After treatment, SGRQ score was decreased in both group, and difference in symptom score was statistically significant between the two groups (t=2.109, P =0.037).The adverse reactions were found in 3 cases in ELP group, and in 2 cases in control group with no statistically significant difference between the two groups [3 (4.8％) vs.2 (3.5％), x2 =0.013, P=0.732].Conclusions Long-term oral administration of eucalyptol-limonene-pinene enteric capsule can significantly decrease exacerbation frequency, improve quality of life, delay the deterioration in lung function, and have good safety in elderly patients with stable COPD.

Objective To clarify the age patterns and types of differences so as to provide reference on prevention and interventions of hand,foot and mouth disease (HFMD) cases,in Hubei province.Methods We collected the HFMD case information of Hubei province from the Chinese National Notifiable Infectious Disease Reporting System in 2009-2015 while the information on pathogens from the laboratory monitoring system of Center for Disease Control and Prevention at all levels in Hubei province.All the data were stratified by age,disease severity,laboratory confirmation status,and serotypes of enterovirus.Results There were 495 783 reported HFMD cases from 2009 to 2015,in Hubei province,of which 1 045 were severe with 99 fatal.The annual notification rate was 1 231.0/106.HFMD cases were concentrated mainly in 0.5-5 year olds,with highest severity and mortality seen in 6-11 month-olds.The predominated pathogen in mild laboratory-confirmed cases each year,in order during 2009-2015 as:EV71,Cox A16,Cox A16,Cox A16,EV71,Cox A16 and other EV.HFMD showed semiannual peaks in April-June,November-December,and with more cases in the even years than in the odd years.Conclusions Children aged 0.5 to 5 years with 6 to 11 month-olds in particular,were the focused groups of attention in Hubei province.Our findings provided evidence for the improvement on monitoring program.Targeted intervention approaches should be strengthened to reduce the mortality and morbidity of HFMD in the province.

Objective:To investigate the factors affecting the time taken for B cell reconstitution after rituximab (RTX) treatment in children with steroid-sensitive nephrotic syndrome.Methods:This was a retrospective cohort study. The clinical data of 42 children with SSNS who received treatment with RTX in Department of Nephrology, Rheumatology and Immunology, Children′s Hospital Affiliated to Zhengzhou University between December 2019 and May 2023 were analyzed retrospectively. The data of demographics, immunosuppressant treatment and laboratory tests such as CD19 +B cell count, urinary protein quantification were collected. The patients were divided into 2 groups, the early B cell reconstruction group and the late reconstruction group based on the average time of B cell reconstruction. A multivariate logistic regression model was used to analyze the factors impacting the timing of B cell reconstruction, and the predictive value of these factors was assessed by plotting the receiver operating characteristic (ROC) curve. Results:There were 42 children, with 35 males and 7 females. They were aged 3.5 (2.2, 5.9) years at the onset of PNS and (8.4±3.3) years at their first RTX treatment. The time for B cell reconstitution was (152±53) d. There were 20 children in the early reconstruction group and 22 children in the late reconstruction group. There were no statistically significant differences (all P>0.05) between the 2 groups in terms of the cumulative dose of steroids within 1 year before receiving RTX infusion (0.29 (0.16, 0.50) vs. 0.29 (0.19, 0.46) mg/(kg·d)), the percentage of children using tacrolimus before RTX (65%(13/20) vs. 45%(10/22)) and cumulative doses (0.04 (0.03, 0.05) vs. 0.03 (0.03, 0.06) mg/(kg·d)), the steroid doses at the time of RTX infusion (0.73 (0.49, 0.90) vs. 0.71 (0.58, 0.89) mg/(kg·d)), the percentage of children using tacrolimus at the initial RTX infusion (50% (10/20) vs. 41% (9/22)) and the doses (0.03 (0.02, 0.04) vs. 0.02 (0.01, 0.04) mg/(kg·d)), the discontinuation time of tacrolimus post-RTX infusion (71 (42, 91) vs. 64 (42, 91) d). A multivariate analysis revealed a correlation ( OR=0.26, 95% CI 0.10-0.68, P=0.006) between B cell count following the second RTX infusion and the time taken for B cell reconstruction. The area under the ROC curve for B cell count after the RTX infusion in predicting the time to B cell reconstruction was 0.89 (95% CI 0.78-0.99, P<0.001) and the cut-off value was 0.925×10 6/L. Conclusions:The time of B cell reconstruction is not influenced by the previous or concurrent use of tacrolimus, regardless of its duration and the dosage of steroid and tacrolimus prior to the RTX infusion. Insteadly, the peripheral blood B cell count (0.925×10 6/L) following the second RTX infusion for SSNS is identified as an independent predictor of reconstruction time, allowing for a more precise prediction and early intervention to maintain disease remission.

Objective:To summarize the clinical and genetic characteristics, treatment and prognosis of four children with Steroid-resistant nephrotic syndrome (SRNS) due to variants of TRPC6 gene. Methods:Clinical data of four children with SRNS admitted to Children′s Hospital Affiliated to Zhengzhou University between May 2020 and August 2022 were collected. Peripheral blood samples were collected from the children and their parents, and whole exome sequencing was carried out. Sanger sequencing was used to verify the pathogenicity of the candidate variants among the children and their parents.Results:All of the four children were found to harbor heterozygous variants of the TRPC6 gene, including c. 523C>T (p.R175W), c. 1327T>A (p.F443I), c. 430G>C (p.E144Q) (unreported previously), and c. 523C>T (p.R175W), which were all missense variants. Two of the children have shown a simple type, whilst two have shown a nephritis type, none had extrarenal phenotype. Comprehensive renal pathology of three children revealed focal segmental glomerulosclerosis (FSGS). Two children were treated with steroids combined with calcineurin inhibitors (CNIs), among whom one showed significant improvement in symptoms. Conclusion:Discoveries of the novel c. 430G>C variant and the new SRNS phenotype of the c. 1327T>A variant have expanded the mutational and phenotypic spectrum of the TRPC6 gene, which has provided a reference for clinical diagnosis and genetic counseling for the families.

Transient receptor potential cation channel 6（TRPC6）gene is mainly expressed in renal podiocytes.Its variation can lead to steroid-resistant nephrotic syndrome（SRNS），and the specific pathogenesis is not clear.These children have poor response to hormones and immunosuppressants，with lack of specific therapeutic drugs，and poor prognosis.In recent years，it has been found that some drugs can slow down disease progression by inhibiting the expression of TRPC6 gene or its downstream signaling pathway，and the discovery of TRPC6 protein-specific blockers may be the hope of treating such children.This review focuses on the pathogenesis of SRNS induced by TRPC6 gene variation in children and the research progress of drug therapy.