Aim To screen out the antigenic sequences from HCV core protein random peptide libraries displayed on phage and to explore a new way to screen the viral antigens. Methods The anti-HCV core antibody-positive serum was used to screen antigenic peptides from the HCV core protein random peptide libraries displayed on phage for 4 rounds. Detection of numbers of positive clones, positive rate of insertion of HCV random DNA and positive rate of hybridization with HCV core probes were used to evaluate the screening effects. The DNA sequences of 7 selected clones with positive hybridization were determined and analysed. Results Six out of 7 sequences are HCV core protein sequences, in which 5 were perfectly displayed,and one was possibly displayed. These sequences included several major HCV core antigenic epitopes. The remaining one was E.coli nrfa gene. Conclusion The phage display technique can be applied to study the viral antigenic peptides with the advantages of simple, accuracy and rapidity.

To improve 3-ketosteroid-△1-dehydrogenase(KSDH) activity and the transformation level for androst-4-ene-3,17-dione, 3-ketosteroid-△1-dehydrogenase gene(ksdD) from Arthrobacter simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55kDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110?0.5mU and 15?0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for steroid medicines production.

Social communication disorders and language disorders are commonly found in children with autism spectrum disorders,and it is also one of the indicators for assessing the severity of the syndrome. This study analyzed the characteristics of language difficulties and social communication disorders in children with ASD. Then,seven evidence-based treatment methods of language rehabilitation for children with ASD were introduced,including Comprehensive Behavioral Treatment for Young Children,Pivotal Response Treatment,Natural Teaching Strategies,Language Training(Production),Scripting,Story-based Interventions and Social Skills Package. The interventions recommended by American Speech-LanguageHearing Association(ASHA) were also introduced. It is of great significance to provide the early intervention of language disorders for children with ASD. Community-based and family-based intervention models should be promoted,and parent training should be actively carried out,so that children with ASD can obtain language rehabilitation during the critical period of language learning.

OBJECTIVE Liver cancer is one of the most common causes of cancer related deaths worldwide, specially, in China. Hinesol, extracted from Atractylodeslance a(Thunb.) DC. has been proved that has anti-cancer effect in leukemia in vitro and in vivo.However,it has been not well under-stood in liver cancer cells.METHODS Cell proliferation,apoptosis,cell cycle and invasion were performed to investigate the anti-liver cancer effect of hinesol in SMMC-7721 and LM3 by MTT assay,flow cytometry and scratch assay.Western blot was used to research the potential mechanism.RESULTS We revealed that hinesol suppresses cell proliferation and invasion,prompts population of G1 phase,induces apop-tosis in dose-dependent manner in SMMC-7721 and LM3 cells.Western blot data showed that hinesol could inhibits the expression of cyclin-D1, Bcl-2 and Bax, and inhibited phosphorylation of MEK and ERK, down-regulated the expressions of NF-κB p65 and phosphor-p65 in nucleus. The results indicated that hinesol reduces cell proliferation via arresting cell cycle at G1 phase and induces apoptosis.Further-more,western blot showed that hinesol inhibited phosphorylation of MEK and ERK,down-regulated the expressions of NF-κB p65 and phosphor-p65 in nucleus.CONCLUSION Our results demonstrate that hinesolreduces cell proliferation via arresting cell cycle at G1 phase and induces apoptosis, it has potent anti-cancer effect against liver cancer cells via down-regulation of MEK/ERK and NF-κB pathway,and indicate that hinesol is a potential liver cancer drug for further research.

The selective side-chain cleavage of phytosterol to 4-androstene-3,17-dione(4-AD)and 1,4-androstadiene-3,17-dione(ADD)by Mycobacterium sp.was described.Because of the similarity in chemical structure between 4-AD and ADD,it is difficult to separate them from the fermentation broth.So far,it has been verified that the ADD can be produced by dehydrogenation of 4-AD.In this reaction,3-Ketosteriod-1-Dehydrogenase(ksdD)plays an important role.The gene knocking out method was used to solve the problem.Partial sequence of ksdD was obtained by PCR which was 631bp in length.Then,a targeting vector pUC19-MK was constructed,which was electroporate into the original strain Mycobacterium neoauru.The method of homologous recombination was used to knock out ksdD gene located in the chromosome of Mycobacterium neoauru.In this way,ksdD would lose its enzyme activity.In the result,5 transformants were screened.The experiments of steroid transformation by the transformants were carried out.The productivity of 4-AD reached 17.52% after 144h,which is 192% higher than the original strain.Meanwhile,the productivity of ADD reached 6.12%,which is 89.9% lower than the original strain.

To improve 3-ketosteroid-△1-dehydrogenase (KSDH) activity and the transformation level for androst-4-ene-3,17-dione,3-ketosteroid-△1 -dehydrogenase gene (ksdD) from Arthrobacter simplex was cloned into plasmid pWB980 and expressed in B. subtilis WB600 under the control of promoter P43. The molecular weight of expressed enzyme was about 55kDa by SDS-PAGE analysis. The activitities assayed by spectrophotometrical method of intracellular and extracellular soluble enzyme were 110 ± 0.5mU and 15 ± 0.6mU per milligram of protein respectively. The transformation rate of androst-4-ene-3,17-dione by the B. subtilis recombinant cells was 45.3%. Compared with Arthrobacter simplex, the enzyme activity of KSDH expressed in B. subtilis was improved about 30 fold, and the transformation level of androst-4-ene-3,17-dione by the B.subtilis recombinant cells was improved about 10 fold. The recombinant B. subtilis cells used in biotransformation of steroids provided a new way for steroid medicines production.

Antibodies, Monoclonal ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; genetics ; metabolism ; Humans ; Immunohistochemistry ; methods ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; diagnosis ; genetics ; metabolism ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND: At present, finite element analysis can be used to judge intertrochanteric fractures, but mostly limited in the distribution of stress. Finite element model of various intertrochanteric fractures has not been reported in detail.OBJECTIVE: To build various types of intertrochanteric fracture models with Hypermesh 14.0 and LS-DYNA software to simulate the falling-induced external force on proximal femur, and to evaluate the effect of models, and to analyze the biomechanical mechanism of intertrochanteric fractures. METHODS: Normal side CT image data of one case of elderly intertrochanteric fracture were collected and imported into Mimics software to establish the proximal femur geometric models, were then analyzed and operated by LZ-DYNA solver after imported into Geomagic studio 2013 and Hypermesh 14.0 for smoothing and meshing. Before analysis, the material parameters were set, the boundary conditions were confirmed, and given the loading parameters. The operating results were checked in Hyper View. RESULTS AND CONCLUSION: (1) The distribution of stress of proximal femur exactly matched to the previous study. EvansⅠtype intertrochanteric fracture model was obtained under continuous shear stresses, and six types of fractures were obtained by adjusting the load. (2) These results manifest that based on the Hypermesh 14.0 and LS-DYNA software, the finite element can well simulate the intertrochanteric fractures, and shear stress plays an important role in intertrochanteric fractures, which can provide experimental basis for the prevention and treatment of intertrochanteric fractures.

OBJECTIVETo determine the relationship between the extent of the damage and clinical data in Alzheimer's disease (AD).

METHODSTwenty-two patients with AD and twenty-two controls received MR-diffusion tensor scanning. The fractional anisotropy (FA) values of white matter in AD patients were measured respectively in parietal lobe and the genu of corpus callosum. Independent-samples t-test for non-paired data was used to test differences between AD and controls for FA values. Correlation analysis was applied to reveal the correlations between FA values in each region and the MMSE, FOM, RVR, BD and DS scores.

RESULTSPositive correlations were found between FA values in left parietal lobe and FOM/DS, and between FA values in genu of corpus callosum and MMSE scores.

CONCLUSIONSIn AD, the MR-DTI can reflect the relationship between the degree of white matter abnormalities and the cognitive impairment.

Aged ; Alzheimer Disease ; diagnosis ; pathology ; psychology ; Anisotropy ; Cerebral Cortex ; pathology ; Cognition ; Diffusion Magnetic Resonance Imaging ; Female ; Humans ; Male ; Middle Aged ; Wechsler Scales

To synthesize and evaluate antimicrobial activity of novel heterocyclic compounds, the corresponding title aroyl ureas have been synthesized by the reaction of 2-amino-5-(3-pyridyl)-1, 3, 4-thiadiazole with aroyl isocyanates. Their antimicrobial activities in vitro were tested by disk diffusion methods and broth microdilution according to M-27A protocol recommended by NCCLS. Twelve new compounds were obtained, and their structures were confirmed by MS, IR, 1H NMR and elemental analysis. The biological screening tests showed that most of the compounds have some antifungal activities in vitro. Aroyl ureas incorporating pyridyl thiadiazole ring may be developed as novel antifungal candidate drugs and are worthwhile to be further studied.
Antifungal Agents ; chemical synthesis ; chemistry ; pharmacology ; Candida albicans ; drug effects ; Microbial Sensitivity Tests ; Molecular Structure ; Phenylurea Compounds ; chemical synthesis ; chemistry ; pharmacology ; Thiadiazoles ; chemical synthesis ; chemistry ; pharmacology