ObjectiveTo study the mechanism of differentiation of mesenchymal stem cells(MSCs) into neuron-like cells in vitro.MethodsMSCs of Wistar rats were separated and cultured, and then induced with DMSO and BHA in vitro. The specific marking proteins of neurons, glia and neural stem cells were detected before preinduction, at 24h after preinduction, at 6h, 24h, and 48h after neuronal induction.ResultsAfter the inducement, many MSCs turned into bipolar,multipolar and taper,and then intersected as network structure. Nestin was strong positive at 6h after neuronal induction, and decreased at 24h, 48h after the induction. NeuN was present at 6 h after neuronal induction, and increased at 24h, 48h after the induction.ConclusionMSCs can be induced into neural stem cells(NSCs) at first, and then differentiate into neuron-like cells in vitro.

OBJECTIVE: To evaluate and compare outcomes effects of Chinese recognition materials in post-lingually deafened patients with cochlear implant. METHOD: Thirty-two post-lingually deafened cochlear implant users participated in the study. Each one of them was given a series of speech recognition tests including the speech recognition score of monosyllable words, spondee words, Chinese BKB sentences and MHINT in quiet, as well as speech recognition threshold of HOPE corpus in babble noise. Speech recognition scores and thresholds were compared among these test materials using histograms , scatter diagrams and statistical methods. RESULT: Recognition scores for spondee words, Chinese BKB sentences and MHINT were affected by ceiling effects, with 1,14 and 4 cases scored 100% respectivelty. Meanwhile, 17, 26 and 14 cases scored more than 85% correspondingly. On the other hand, speech recognition scores for monosyllable words in quiet and speech recognition threshold for HOPE corpus in babble noise were not affected. CONCLUSION For a considerable part of post-lingually deafened patients using cochlear implant, some Chinese speech recognition materials have demonstrated different degrees of ceiling effects. While speech recognition scores for monosyllable words in quiet and recognition threshold for HOPE corpus in noise can evaluate the performance of speech recognition abilitymore objectively.
Cochlear Implantation ; Cochlear Implants ; Humans ; Language ; Noise ; Speech Perception ; Treatment Outcome

Objective To explore the current status of home-based rehabilitation of patients who accepted reconstruction of anterior cruciate ligament in the early stage,so as to analyze its correlation with their joint function recovery in early.Methods Fifty-four consecutive patients underwent arthroscopic reconstruction of anterior cruciate ligament were recruited.A self-designed questionnaire was used to collect demographic data,the implementation of rehabilitation,concerns about the exercises,and function status of the knee were observed in outpatient after their surgeries two weeks.Results Only 25 patients (46.3％) had a satisfied function recovery of the knee. Control rate of muscle activity was 33.3％,and control rate of range of movement exercises was 29.6％. Varying degrees of concern to rehabilitation was founded in patients,and the most concern was afraid of pain (55.6％),improper practice due to lacking of guiding (51.9％),hang on due to boring during rehabilitation process (37.0％). Early functional recovery of patients was influenced by the control rate of rehabilitation.Conclusions Current status of early functional recovery of patients who accepted reconstruction of anterior crnciate ligament in the early stage is far from satisfied.More attention should be paid to home-based rehabilitation.It should be improved by strengthening the guidance of rehabilitation in-hospital and follow-up in outpatient.

The systematic treatment based on gemcitabine plus cisplatin is recommended as the current standard chemotherapy for unresectable or metastatic biliary tract cancers.However,the exact benefits from the recognized regime are still dismal.We thus elicit this study in an attempt to analyze whether targeted therapy coupled with various chemotherapy could produce improvement of survival benefits.The clinical trials were searched electronically from databases till July 2016 published in English and Chinese.Nine hundred and sixty-four patients from 7 trials were identified in our analysis.The overall analysis achieved a significantly higher overall response rate (ORR) among the patients treated with targeted drugs plus chemotherapy than chemotherapy alone (OR=1.87;95％ CI:1.37-2.57;P=0.000),but failed in the overall progression-free survival (PFS) [mean difference (MD)=0.63;95％ CI:-0.45-1.72;P=0.26] and overall survival (OS) (MD=-0.67;95％ CI:-2.54-1.20;P=0.49).In the sub analysis,better ORR was obtained with the addition of EGFR (OR=1.75;95％ CI:1.20-2.56;P=0.004) and VEGFR (OR=2.5;95％ CI:1.28-4.87;P=0.007) targeted therapy.Furthermore,the sub analysis of EGFR target showed an significant improvement on PFS (MD=l.36;95％ CI:0.29-2.43;P=0.01).No significant differences were observed in the incidences ofneutropenia (OR=1.37;95％ CI:0.89-2.12),thrombocytopenia (OR=l.40;95％ CI:0.83-2.39),anemia (OR=l.21;95％ CI:0.62-2.38),peripheral neuropathy (OR=1.52;95％ CI:0.81-2.88),increased AST/ALT (OR=l.40;95％ CI:0.82-2.39) as well as fatigue (OR=1.65;95％ CI:0.96-2.84) in either of the treatment groups.In conclusion,better ORR associated with chemotherapy combined with targeted therapy (both targeting EGFR and VEGF) is found in the present mcta-analysis without the cost of increased unacceptable toxicities,but regretfully not for the OS.The sub-analysis of targeting EGFR instead of VEGF obtains a superior PFS.Otherwise,there is no statistically significant difference in the overall PFS between the combination regime and chemotherapy alone.Given the paucity of favorable data,we need further studies to characterize optimal targeted agents to confirm the potential value to biliary tract cancer.

To characterize the genomic sequence and arrangement of WU polyomavirus (WU virus) identified in clinical specimens collected from children with acute respiratory infections in Beijing, China, the sequences of capsid proteins VP1, VP2, and the large tumor antigen (LTAg), as well as the 5'-terminal sequence of WU virus, were amplified from the clinical specimen with ID number of BJF5276 which was determined as WU virus positive by PCR amplification. The PCR amplicons were sequenced, and genomic sequence analysis was performed by using the software DNAStar. In addition, VP2 coding-region sequences were amplified from other 21 clinical specimens identified as WU virus positive to investigate the gene diversity of WU virus. The genomic sequence of WU virus BJF5276 with accession number of HQ218321 in GenBank was 5,229 base pairs in length with 3 major coding domain sequences (CDS) sited on one strand coding for capsid proteins VP2, VP3 and VP1, and two CDS sited on the complementary strand coding for small tumor antigen (STAg) and LTAg; These 22 VP2 CDS sequences including 5 sequences submitted to GenBank were compared with 64 corresponding sequences downloaded from GenBank by MegAlign of DNAStar software, indicated that these sequences coming from children in Beijing shared high homology (over 98.8%) with those from GenBank. Phylogenetic analysis of these VP2 CDS by using Neighbor-joining (NJ) analyses with 2,000 bootstraps (Mega 4.0) showed that 20 sequences out of 22 belonged to clade Ia, and other 2 of them belonged to clade III, including 1 clustered in IIIa and 1 in a novel cluster proposed as IIIc. In conclusion, the genomic sequence of WU polyomavirus detected from clinical specimens from children in Beijing is closely related to other WU polyomaviruses in the feature of genomic coding region arrangement. Overall variation of VP2 CDS was very low, and there were different clades circulating in Beijing with a dominant clade Ia, which is different from dominated Ib circulating in other parts of the world reported previously, and a novel clade IIIc was proposed.
Acute Disease ; Child, Preschool ; China ; Female ; Genome, Viral ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Polyomavirus ; classification ; genetics ; isolation & purification ; Respiratory Tract Infections ; virology ; Viral Proteins ; genetics

BACKGROUNDThe DFNB1 locus, which contains the gap junction beta-2 (GJB2) and gap junction beta-6 (GJB6) genes, plays a key role in the nonsyndromic and sporadic hearing impairment. Mutations of DFNB1 result in autosomal recessive nonsyndromic hearing impairment (ARNSHI). Previous researches have identified mutations in GJB2 and GJB6, but single nucleotide polymorphisms (SNPs) of DFNB1 locus have not been studied. So we chose five SNPs to evaluate whether there is difference between deafness people and normal-hearing people in Han Chinese.

METHODSFive SNPs in the DFNB1 region were examined using a case-control association study between cases with sporadic hearing impairment and controls with normal hearing. The HWEsoft and SHEsis softwares were used to analyze the results.

RESULTSSingle-locus association analysis showed a positive association for three SNPs: rs9315400, rs2274084 and 235delC. When we compared the distributions of the haplotypes, we also found significant differences between cases and controls in the haplotype combination of rs2274084 and rs2274083 (chi(2) = 12.978, df = 3, global P = 0.004719).

CONCLUSIONSThe haplotypes composed of rs2274084 and rs2274083 suggested that C-C may be a risk haplotype for the sporadic hearing impairment while T-T may be protective against hearing impairment. From that point of view, we can conclude that the SNPs of DFNB1 locus also plays an important role in sporadic hearing impairment cases.

Adolescent ; Child ; Connexin 26 ; Connexin 30 ; Connexins ; genetics ; Female ; Haplotypes ; Hearing Loss ; genetics ; Humans ; Male ; Polymorphism, Single Nucleotide

The aim of this study was to obtain the capsid protein VP2 of human bocavirus (HBoV) identified in Beijing recently and construct virus-like particles (VLPs) in insect cells for further study of this virus. The full-length VP2 gene of HBoV from BJ3722 was inserted into the baculovirus expression transfer vector (pFastBac 1) to obtain the recombinant Bacmid, and generation of recombinant baculoviruses was followed by transfection of the recombinant Bacmid into insect cells. Then the recombinant VP2 protein was recognized by SDS-PAGE using Coomassie-blue staining and Western blot using hyper-immune serum against VP2 of HBoV from rabbit. The recombinant baculoviruses were harvested and amplified to gain large amounts of viruses with high titers to infect insect cells at a multiplicity of infection (MOI) of 0. 5. After 7-10 days or 4-5 days of the infection, the supernatants of culture or the cell lysates treated with lysing solution were harvested, and ultracentrifuged twice through 40% sucrose cushion to obtain purified VLPs, which were followed by Western blot and IFA for VLPs' composition and specificity analysis, by electron microscopy for VLPs' morphologic structure. The recombinant VP2 protein with molecular weight of approximately 61 kD expressed in recombinant baculoviruses was recognized by SDS-PAGE using Coomassie-blue staining and Western blot. The presence of VP2 on VLPs was demonstrated by Western blot and IFA from samples collected during the purification of VLPs from the supernatants of culture or the cell lysates, and the expression of VP2 in insect cells led to the formation of VLPs which formed the typical icosahedral appearance of parvoviruses with a diameter of approximately 20 nm. In conclusion, the recombinant baculoviruses were constructed, the HBoV VP2 protein was expressed in insect cells with high specific antigenicity and VLPs was formed successfully.
Animals ; Blotting, Western ; Capsid Proteins ; genetics ; metabolism ; Cell Line ; Electrophoresis, Polyacrylamide Gel ; Fluorescent Antibody Technique, Indirect ; Human bocavirus ; genetics ; metabolism ; Microscopy, Electron, Transmission ; Polymerase Chain Reaction ; Spodoptera ; Virion ; genetics ; metabolism ; ultrastructure

Dioscin has a wide range of biological effects and broad application prospects. However the studies concerning the toxicology and mechanism of dioscin is small. This article is to study the hepatotoxicity of dioscin and the effect of dioscin treatment on expression of aryl hydrocarbon receptor (AhR) mRNA and CYP1A mRNA and protein in HepG2 cells in vitro. Dioscin 0.5-32 µmol · L(-1) exposed to HepG2 cells for 12 h, cell viability was examined by CCK-8 assay and the release rate of lactate dehydrogenase (LDH) was to evaluate cell membrane damage. HepG2 cells morphologic changes were quantified by inverted Microscope, and the effect on production of reactive oxygen species (ROS) was detected by flow cytometry. The mRNA expression of CYP1A and AhR was evaluated by RT-RCR. The protein expression of CYP1A1 was detected by western blot. The cell viability was significantly inhibited after HepG2 cells were exposed to dioscin 0.5-32 µmol · L(-1). Compared with the control, the LDH release rate and ROS were significantly increased. The expression of CYPlA and AhR mRNA was increased. The expression of CYP1Al protein was increased after dioscin treatment, and resveratrol, an AhR antagonist, could downregulate the expression of CYP1A1. It follows that large doses dioscin has potential hepatotoxicity. The possible mechanism may be dioscin can active aryl hydrocarbon receptor (AhR) and induce the expression of CYP1A.
Cell Survival ; drug effects ; Chemical and Drug Induced Liver Injury ; etiology ; Cytochrome P-450 CYP1A1 ; genetics ; Diosgenin ; analogs & derivatives ; toxicity ; Hep G2 Cells ; Humans ; L-Lactate Dehydrogenase ; secretion ; RNA, Messenger ; analysis ; Reactive Oxygen Species ; metabolism ; Receptors, Aryl Hydrocarbon ; genetics

OBJECTIVETo investigate the life style, genetic and occupational risk factors of metabolic syndrome (MS) among policemen.

METHODS1:4 matched case-control study was used, based on physical examination data of Tianjin Policemen in 2010, 708 patients with MS were randomly selected as cases, which were matched with 2832 healthy controls on the basis of sex and age (+/- 1 year). An epidemiological investigations on the past exposure status of several possible risk factors was conducted, and the data were analyzed with conditional logistic regression.

RESULTSFifteen factors related to exposure were identified for MS through univariate conditional logistic regression analysis. Multivariate conditional logistic regression analysis suggested that, seven factors, such as family history of hypertension (OR = 2.406, 95% CI: 1.946-2.975), family history of diabetes (OR = 1.301, 95% CI: 1.043-1.623), smoking (OR = 1.357, 95%CI: 1.010-1.823), snoring (OR = 1.268, 95% CI: 1.043-1.543), work intensity (OR = 4.603, 95% CI: 3.767-5.623), occupational stressful events (OR = 1.524, 95% CI: 1.209-1.922), security policemen (OR = 1.453, 95% CI: 1.127-1.872) and criminal investigation policemen (OR = 2.792, 95% CI: 2.168-3.596), could significantly increase the risk of disease development, but dairy products (OR = 0.782, 95% CI: 0.619-0.989) was a protect factor for MS. The results from population attributable risk factors analysis showed that the control of smoking, snoring, work intensity, occupational stressful events can decreased the risk of MS to 16.26%, 11.71%, 56.87% and 8.97%, respectively.

CONCLUSIONMetabolic syndrome has became a significant public health problem among policemen, it's necessary to take measures on life style, occupational risk factors for reducing the incidence of MS, and improving the health level among policemen.

Adult ; Case-Control Studies ; Factor Analysis, Statistical ; Humans ; Logistic Models ; Male ; Metabolic Syndrome ; epidemiology ; genetics ; psychology ; Middle Aged ; Occupational Health ; Police ; Risk Factors ; Young Adult

OBJECTIVETo investigate molecular epidemiologic features of rotaviruses circulating in Shanghai, China.

METHODSStool samples were collected from 1230 hospitalized children with community-acquired and nosocomially acquired diarrhea in Children's Hospital Affiliated to Fudan University between November 1, 1999 and December 31, 2001. Polyacrylamide gel electrophoresis (PAGE) was used to detect rotavirus genomic RNA and identify electropherotypes of group A rotavirus RNAs. Reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify full length VP7 gene and dot blot hybridization was performed to identify rotavirus G serotypes using digoxigenin-labelled variable regions from VP7 genes as probes. These probes were amplified by PCR from recombinant plasmids containing full length G1, G2, G3 and G4 VP7 genes from rotavirus field strains detected in Beijing and digoxigenin labelled dUTP was integrated into the PCR products. The Kruskal-Wallis analysis of variance was employed to analyze whether there were significant differences in variables.

RESULTSOut of 1230 samples investigated, 493 (40.1%) were group A rotavirus gene positive by PAGE, among which 397 (80.5%) showed long electropherotypes, 55 (11.2%) showed short electropherotypes, 18 (3.7%) showed mixed electropherotypes which suggested that the children were co-infected by rotaviruses with different electropherotypes, 23 (4.7%) were non-typable because of degradation of some of the genomic RNA fragments. No group B or group C rotavirus was found. RT-PCRs were performed for 328 fecal specimens containing sufficient rotavirus RNAs and VP7 gene products were obtained from 254 (77.4%) samples. Dot blot hybridization showed serotype G1 accounted for 55.5% (141) of these samples, serotype G3 accounted for 27.6% (70), serotype G2 accounted for 9.4% (24), co-infection by 2 rotaviruses with different G types accounted for 6.3% (16), only 1 G4 was detected and 2 were non-typable. The genomic RNA patterns of all G2 strains were short and those of G1, G3 and G4 strains were long. There were no statistically significant differences for age distribution and clinical manifestations among those infants and children infected by rotaviruses with different G serotypes.

CONCLUSIONGroup A rotavirus is the major pathogen for diarrhea in infants and children in Shanghai during the period of Nov. 1999 to Dec. 2001. Rotaviruses with long electropherotype were dominant during these years. Serotypes G1 to G3 constituted 98.8% of all 254 strains tested, and G1 was the most common serotype followed by G3 and G2, whereas serotype G4 was seldom found. Some of the children were co-infected by rotaviruses with different G serotypes. Clinical manifestations were not related to the infecting rotavirus with different G serotypes.

Age Factors ; Antigens, Viral ; Capsid Proteins ; genetics ; metabolism ; Child, Preschool ; China ; epidemiology ; Data Collection ; Dysentery ; epidemiology ; etiology ; Electrophoresis, Polyacrylamide Gel ; Feces ; virology ; Female ; Humans ; Infant ; Male ; RNA, Viral ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Rotavirus ; classification ; genetics ; Rotavirus Infections ; complications ; epidemiology ; virology ; Serotyping