OBJECTIVETo study the autophagy activity between rat bone marrow stem cells (BMSCs) neural differentiation in order to explore the mechanism involve in this process.

METHODSBMSCs were passed by 3 generation, then was induced with the revulsant 2% (DMSO) + 200 µmol/L (BHA), NSE expression was detected by immunocytochemical stain, the mRNA expression of autophagy associated genes L3B, Beclinl, Atg5, Atg7, Atg10 were detected by RT-PCR, the autophagy protein LC3B was examined by Western blot and flow cytometry analysis.

RESULTSBMSCs were passed by 3 generation, the purity of BMSCs could reach more than 90%, the morphology of cells were like fibroblasts, after the revulsant 2% DMSO + 200 µmol/L BRA induced, cells were extended long neurites, like nerve cells, positive rate of NSE staining was (83±5) %, RT-PCR results showed that the expression of autophagy associated genes LC3B, Beclinl, Atg5, Atg7 Atg0 were rised after BMSCs neural differentiation, Western blot analysis showed that the LC3B-II protein expression was increased after neural differentiation and the MFI of L3B was highten by flow cytometry.

CONCLUSIONAutophagy is increased after rat BMSC neural differentiation.

Animals ; Autophagy ; Cell Differentiation ; Cells, Cultured ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; Neurons ; cytology ; Rats

OBJECTIVETo observe the clinical efficacy on pain in postmenopausal osteoporosis treated with the warm acupuncture therapy and discuss its effect mechanism.

METHODSNinety cases of postmenopausal osteoporosis were randomized into a warm acupuncture group, an electroacupuncture group and a medication group, 30 cases in each group. In the warm acupuncture group and the electroacupuncture group, Dazhu (BL 11), Shenshu (BL 23) and Xuanzhong (GB 39) were selected bilaterally and stimulated with the warm acupuncture and electroacupuncture therapies respectively, once a day for 30 days totally. In the medication group, caltrate-D tablets were prescribed, 600 mg, once a day for 30 days totally. The changes in the bone density T value, visual analogue scale (VAS) score, serum insulin like growth factor 1 (IGF-1), interleukin 6 (IL-6) and tumor necrosis factor (TNF-alpha) were observed before and after treatment in the three groups.

RESULTS(1) The bone density T value in the patients of postmenopausal osteoporosis did not change obviously after 30 days treatment with the three therapies; (2) VAS score was all reduced after treatment, in which, the result in the warm acupuncture group was the most obvious (6.73 +/- 0.24 before treatment vs 4.43 +/- 0.26 after treatment). The value after treatment in the warm acupuncture group was different significantly as compared with the electroacupuncture group (5.13 +/- 0.31) and the medication group (5.17 +/- 0.33, both P < 0.05). (3) The level of serum IGF-1 was improved after treatment in the warm acupuncture therapy [(119.5 +/- 20.1) ng/mL before treatment vs (156.5 +/- 23.9) ng/mL after treatment], which was more apparent as compared with the electroacupuncture group [(136.3 +/- 24.5) ng/mL] and the medication group [(127.7 +/- 22.1) ng/mL, all P < 0.05]. Concerning to reducing the levels of IL-6 and TNF-alpha in serum, the results in the warm acupuncture group were superior to the other two groups (all P < 0.05).

CONCLUSIONThe warm acupuncture therapy achieves the significant efficacy on pain in postmenopausal osteoporosis, which could be related to increasing the level of IGF-1, decreasing the levels of IL-6 and TNF-alpha, promoting bone formation and inhibting bone absorption.

Acupuncture Therapy ; methods ; Bone Density ; Female ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Interleukin-6 ; blood ; Middle Aged ; Osteoporosis, Postmenopausal ; blood ; physiopathology ; therapy ; Pain Management ; Tumor Necrosis Factor-alpha ; blood

In order to demonstrate PTB bind to HPRE,reverse transcription,PCR-mediated detection,were used.HepG2.2.15 cell line and HBs-HPRE transient expression cells were adopted to identify PTB function in HBV life cycle.The results showed that PTB could directly bind to HPRE RNA.Functional analysis indicated that PTB could inhibit the expression of HBs antigen and this inhibition was in a dose-dependent manner in HepG2.2.15 cells.Higher expression of HBs in cells transfected pcDNA3-HBs-HPRE comparing with pcDNA3-HBs,and this high expression could also be inhibited by PTB.The data demonstrated that PTB inhibits HBs expression by interacting with HPRE.

OBJECTIVETo investigate the effects of formaldehyde inhalation on the morphological damage, and Glu, GABA and NOS contents in olfactory bulb and hippocampus of rats.

METHODSTwenty SD rats were equally divided into two groups: rats in the control group inhaled fresh air, while the animals in experimental group were exposed to the air containing formaldehyde (12.5 mg/m(3), 4 h/d) for 7 days. Then rats were sacrificed and frozen sections of olfactory bulb and hippocampus were prepared. The morphological changes were examined and the Glu, GABA and NOS contents were detected using Nissl-staining, immunohistochemistry and Western blot, respectively.

RESULTCompared with the control group, there was a significant confusion and shrink of neuron morphology in experimental group, the number and staining intensity of Glu and NOS positive cells and protein contents were reduced. The protein expression of GABA was also decreased in the formaldehyde group.

CONCLUSIONFormaldehyde inhalation can cause a severe morphological damage of olfactory bulb and hippocampus in SD rats,which may further impair memory and learning ability through the reduction of Glu, GABA and NOS expression.

Animals ; Formaldehyde ; toxicity ; Glutamic Acid ; metabolism ; Hippocampus ; drug effects ; metabolism ; pathology ; Inhalation Exposure ; Learning ; drug effects ; Neurons ; drug effects ; metabolism ; pathology ; Nitric Oxide Synthase ; metabolism ; Olfactory Bulb ; drug effects ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; gamma-Aminobutyric Acid ; metabolism

OBJECTIVETo observe whether H. pylori inoculated by oral route could arrive in livers and cause liver inflammation as an independent etiological factor.

METHODSC57BL/6 mice were orally inoculated with H. pylori SS1 strains and fed for 8 months. H. pylori colonization and pathologic consequences were studied in the liver and gallbladder tissues of the mice; the blood, liver tissue and gastric mucosa were obtained and cultured for H. pylori growth; The bacterial DNA extracted from the liver, bile and blood was examined by nested PCR for H. pylori genes. 16S rRNA PCR amplicons were sequenced and compared with the sequencing results of 16S rRNA PCR amplicons of the bacteria cultured from gastric mucosa and the inoculated H. pylori SS1.

RESULTSThe bacterial DNA extracted from the liver, bile and blood of the infected mice was detected for H. pylori genes by nested PCR. Six of the 15 samples were positive (40%) in the liver, 6 of 10 samples in the bile (60%), and 2 of 10 samples in the blood (20%). Sequencing results of 16S rRNA PCR products of the livers showed 100% homogeneity when compared with the cultured H. pylori from gastric mucosa and inoculated H. pylori SS1. H. pylori was found in 4 liver tissues of the 15 infected mice (26.7%) and 6 in the gallbladders (40%). Infiltrations of lymphocyte cells along hepatic sinusoids and a lower degree infiltration around interlobular arteries and veins were observed; ballooning degeneration was also observed in some hepatocytes.

CONCLUSIONH. pylori inoculated by oral route could arrive in the liver and cause inflammation as an independent etiological factor. The routes which the microorganisms took to reach the livers may involve hematogenous and/or biliary system dissemination.

Animals ; Helicobacter Infections ; Helicobacter pylori ; pathogenicity ; Liver ; microbiology ; Male ; Mice ; Mice, Inbred C57BL ; Rats

OBJECTIVETo assess the prevalence of pre-hypertension and hypertension as well as the association with overweight/obesity in Shanghai adolescents.

METHODSSchool children aged 11 - 17 years in four schools in Shanghai were included in this survey. All students were visited in May 2010 at school by trained nurses or physicians who administered a questionnaire and carried out anthropometric measurements. Anthropometric measurements included height, weight, heart rate and blood pressure. Repeat measurements were performed in school children with elevated blood pressure within one month. The pre-hypertension and hypertension was defined on the basis of the 2004 National High Blood Pressure Education Program Working Group definitions. The overweight and obesity was defined using cutoff points recommended by Working Group of Obesity, China (WGOC).

RESULTSThere were 4175 school children aged 11 - 17 years [mean (15.0 ± 1.9) years, 52.3% (n = 2183) girls and 47.7% (n = 1992) boys]. There were 72.5% (n = 3025) participants with normal blood pressure [ girls (n = 1666) 76.3% and boys (n = 1361) 68.3%], 18.0% (n = 750)participants with pre-hypertension [14.2% (n = 310) for girls and 22.1% (n = 440) for boys], 8.3% (n = 346) participants with stage 1 hypertension [8.2% (n = 179) for girls and 8.3% (n = 165) for boys] and 1.3% (n = 54) participants with stage 2 hypertension [1.3% (n = 28) for girls and 1.3% (n = 26) for boys]. The prevalence of overweight and obesity was 6.7% (n = 147) and 3.0% (n = 66) for girls and 13.7% (n = 273) and 6.3% (n = 125) for boys. After adjusting for gender, age, parental history of hypertension and physical activities, multivariable logistic regression analysis showed that the overweight [adjusted odds ratio and 95% confidence interval: 1.42 (1.16 - 1.75)] and obesity [adjusted odds ratio and 95% confidence interval: 2.35 (1.78 - 3.11)] were independent predictors of elevated blood pressure in this cohort.

CONCLUSIONSThe prevalence of elevated blood pressure is common in adolescents in Shanghai. Overweight and obesity are closely associated with the elevated blood pressure in this cohort.

Adolescent ; Child ; China ; epidemiology ; Epidemiological Monitoring ; Female ; Humans ; Hypertension ; epidemiology ; Male ; Obesity ; epidemiology ; Overweight ; epidemiology ; Prevalence ; Risk Factors

OBJECTIVETo screen and identify proteins that interact with hepatitis C virus NS3 by means of T7-phage display system.

METHODSHepatitis C virus NS3 was expressed by prokaryotic expression and used as a selected molecule to biopan the T7 select human liver cDNA library; the selected positive clones were identified using DNA sequencing and analyzed with BLAST program in GenBank.

RESULTSAfter BLAST analysis in all the positive clones, the proteins which interacted with the hepatitis C virus NS3 were found to be serpin peptidase inhibitor, clade A, member 1 (SERPINA1) and cyclophilin-LC.

CONCLUSIONT7-phage display system is a convenient, rapid and effective method for screening interacting proteins. The proteins thus selected will provide an important means for studying the pathogenesis and carcinogenesis of HCV.

Cell Line ; Gene Library ; Hepacivirus ; metabolism ; Humans ; Peptide Library ; Protein Interaction Mapping ; methods ; Viral Fusion Proteins ; genetics ; isolation & purification ; metabolism ; Viral Nonstructural Proteins ; genetics ; metabolism

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Cytotoxicity, Immunologic ; Dendritic Cells ; physiology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; Immunophenotyping ; K562 Cells ; Leukemia ; drug therapy ; immunology ; Lymphocyte Culture Test, Mixed

OBJECTIVETo establish a stable cell line that can replicate hepatitis B virus (HBV) DNA carrying the reverse transcriptase sequence derived from a clinical isolate.

METHODSNested PCR was used to amplify the HBV DNA fragment from the serum. The fragment was cloned into a plasmid that can support HBV replication in vitro by fragment substitution reaction (FSR), followed by the cloning of the neomycin expressing fragment downstream from HBV DNA. G418 selection was conducted after the transfection of HepG2 cells with the recombinant DNA. Real-time PCR and enzyme linked immunosorbent assay (ELISA) were used to screen stable cell lines that can replicate HBV DNA, and the replication of HBV DNA by the cell line was confirmed by using Southern blot analysis.

RESULTSFragment nt55-1654 amplified from the serum DNA was substituted to the plasmid pLL, generating the plasmid p11. The neomycin fragment was cloned into p11, leading to the plasmid p11-neo, and p11-neo was confirmed to be HBV-replication-competent. A stable cell line named 3-10 that can replicate HBV DNA was obtained.

CONCLUSIONSA stable cell line was established that can replicate HBV DNA carrying the reverse transcriptase sequence derived from a clinical isolate. Real-time PCR plus ELISA may help to rapidly screen out stable cell lines replicating HBV DNA.

Cell Line ; Cloning, Molecular ; DNA Replication ; DNA, Viral ; biosynthesis ; Genetic Vectors ; Hep G2 Cells ; Hepatitis B virus ; genetics ; Hepatocytes ; cytology ; virology ; Humans ; Plasmids ; RNA-Directed DNA Polymerase ; genetics ; Virus Replication ; genetics

Objective To analyze the podiatric conditions of athletes with intellectual disabilities.Methods From May eleventh to 17th, 2015,779 athletes who participated in the sixth National Special Olympics were enrolled partly.Their lower limbs and feet were evaluated with Fit Feet Healthy Athletes Screening Form.Results Common podiatric conditions found in Special Olympics athletes included flat feet (52.2% in left side and 51.7% in right side), over pronation (56.1% in left side and 56.9% in right side), callus (44.7%), wrong nail cut (46.6%),ingrown nail(21.1%),and cross over toes(6.8%),etc.Conclusion Special Olympics athletes have podiatric structural,functional, nail and skin problems,which may relate to intellectual disabilities.