Objective To evaluate the strictness and self-consistency of the charge nurses in the nurses examination using venipunctnre trocar operation as research object.Methods FACETS,the polyhedral Rasch software was used in the examination.Results The strictness and self-consistency of charge nurses were evidently different.Conclusions The variable strictness and self-consistency of the charge nurses in the examination leads to unreliability of the results.Developing a scientific and reliable evaluation system is essential to improve the ability of the teaching nurses and the student nurses as well as the hospital nursing quality.

Objective To investigate the inhibitory effects of overexpression of PTEN on renal epithelial-mesenchymal trans-differentiation induced by TGF-β1,and the signaling transduction mechanism.Methods HKC cells were transfected with GFP-PTEN via lipofectAMINE2000.The efficiency of transfection was detected by fluorescence microscope.The expression of PTEN protein and mRNA in the translected cells was detected by Western blot and RT-PCR respectively.The experiment was divided into four groups:normal group,TGF-β1 stimulation group,GFP-PTEN+TGF-β1 group and empty vector+TGF-β1 group.The expression of E-cadherin,a-SMA,Akt and p-Akt was detected by Western blot.Results Most ceils transfected with GFP-PTEN expressed GFP.The expression of PTEN protein and mRNA was strongly increased when HKC cells were transfected with GFP-PTEN(all P<0.05).In both TGF-β1 stimulation group and empty vector+TGF-β1 group,the expression level of E-cadherin was lower(all P<0.05),while that of p-Akt and a-SMA was higher than in normal group(both P<0.05).The expression level of p-Akt and a-SMA in GFP-PTEN+TGF-β1 group was Iower(both P<0.05),while that of E-cadherin was higher than in TGF-β1 stimulation group and empty vector+TGF-β1 group(both P<0.05).The expression of Akt was similar in the four groups.Conclusion Overexpression of PTEN can inhibit renal epithelial-mesenehymal trans-differentiation induced by TGF-β1 through suppressing the activation of PI3K/Akt signal pathway.

ObjectiveTo study the morphology, normal values and lateral asymmetry of Chinese calcarine sulcus on MRI. Methods High-resolution and transverse MRI were obtained from 40 female volunteers. Brainvisa software was used to reconstruct the calcarine sulcus and measure its average length, depth and width automatically. Results The posterior branch of calcarine sulcus showed six types in the median sagittal plane: bifurcation(32.50%), single peak(25.00%), flat (16.25%), S-shaped (15.00%), double peak(7.50%) and other shape (3.75%); its location had three types: inferior(72.50%), middle(21.25%)and superior(6.25%). The depth of left calcarine sulcus was (15.24±2.67)mm, and the right one was (16.97±3.25)mm, which revealed great statistical significance (P＜0.000 1). The width of left calcarine sulcus was (3.14±0.91)mm, and it was (3.19± 0.83)mm in the right side. The bottom length of calcarine sulcus: the left was (86.47±16.85)mm, the right was (83.62±17.10)mm. The top length of calcarine sulcus: the left was (70.52±12.40)mm, the right was (64.90±15.17)mm. There were not statistical significance in width, bottom length and top length between left and right calcarine sulci. More than half of the end part of calcarine sulci turned to the lateral surface of cerebral hemisphere.Totally 63 cases (78.75%) were found with prominent calcar avis. Conclusion Significant difference of depth between left and right calcarine sulcus of female was found. Three-dimensional reconstruction is an effective method to study the anatomy of calcarine sulcus.

Objective To construct eukaryotic enhanced green fluorescent protein (EGFP) expressing recombinant plasmid, pEGFP-C1-hnRNP A1, which contains coding sequence of human hnRNP A1 (heterogeneous nuclear ribonucleo-protein A1), and to perform cellular localization analysis of EGFP tagged hnRNP A1 under stress. Methods Total RNA was isolated from HeLa cell used for synthesis of first-strand cDNAs using reverse primers that are specific for the 3′-un-translated region of hnRNP A1. hnRNP A1 gene fragments were then amplified by touch-down PCR from those cDNAs and inserted into pEGFP-C1 fluorescent bearing vector through EcoRⅠ/BamHⅡdouble enzyme digestion and T4 DNA Ligase connection. The recombinant pEGFP-C1-hnRNP A1 plasmid was transfected into HeLa cells and green fluorescent tagged fusion proteins was examined by Western blot and confocal fluorescence microscopy. Co-localization of EGFP-hnRNP A 1 with poly (A)+mRNA (the marker of the stress granules), or DCP1a (the marker of processome) were detected by RNA fluores-cence in situ hybridization and immunofluorescence. Results The pEGFP-C1-hnRNP A1 was sequenced and digested cor-rectly by restriction single/double enzyme. The green fluorescent fusion protein was also detected in transfected HeLa cell by Western blot and confocal fluorescence microscopy. EGFP-hnRNP A1 co-localizes with poly(A)+mRNA, but not DCP1a. Conclusion Recombinant eukaryotic plasmid of pEGFP-C1-hnRNP A1 was constructed successfully and expressed effec-tively. EGFP tagged hnRNP A1 takes part in forming stress granules.

Aged ; Carcinoma, Small Cell ; genetics ; Case-Control Studies ; Cytochrome P-450 CYP2E1 ; genetics ; Female ; Humans ; Lung Neoplasms ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

Suspension cultures cell of Sorbus aucuparia (SASC) was used as materials, the changes of physiological and biochemical indexes of SASC after treatment with yeast extract (YE) were detected, and the synthetic mechanism of secondary metabolites in SASC treated with YE was preliminarily explored. The results were as follows: under the assay conditions, SASC was induced to synthesize five biphenyl compounds, and these compounds content changed differently with induction time prolonging; YE treatment inhibited cell growth, the culture medium pH was gradually reduced after treatment; water-soluble protein content showed a trend of slow decline, which was significantly increased in YE treatment group (YE group) compared with the control group (CK group), the maximum relative content was 147.76% in contrast with CK group; both YE group and CK group were extracellular Ca2+ flow influx, but the YE group flow was significantly slow than CK group. The results indicate that YE induced the cells in a stress state, which was not conducive to the growth of cells and forced the cells to synthesize biphenyl compounds against external stress; water-soluble protein may serve as intracellular enzymes involved in the synthesis of compounds regulation; Ca2+ may as signal molecule mediate cell signal transduction respond to YE stress.
Cell Culture Techniques ; instrumentation ; methods ; Culture Media ; chemistry ; metabolism ; Saccharomyces cerevisiae ; chemistry ; Secondary Metabolism ; Sorbus ; growth & development ; metabolism

OBJECTIVETo compare the shear bond strength of the fractured anterior teeth reattached by two different adhesive materials.

METHODSForty crown fractured anterior modes were divided into two groups randomly, with 20 in each group. Group A were reattached by Clearfil SE Bond and Clearfil AP-X, while group B were reattached by Clearfil S3 Bond and Clearfil AP-X. Then the specimens were submitted to an axial compression test in a universal testing machine until tooth fractured. The strength was recorded.

RESULTSThe mean shear bond strength of group A and group B was (324.32 +/- 65.91) N and (263.08 +/- 55.88) N, separately. The mean shear bond strength of group A was statistically higher than group B(t = 3.17, P = 0.000).

CONCLUSIONThe shear bond strength of two-step adhesive Clearfil SE Bond is higher than one-step adhesive Clearfil S3 Bond for the reattachment of fractured anterior teeth.

Adhesives ; Composite Resins ; Dental Bonding ; Dental Stress Analysis ; Dentin-Bonding Agents ; Humans ; Methacrylates ; Resin Cements ; Shear Strength ; Tooth Fractures

OBJECTIVETo clone a novel gene expressed specifically in human embryonic stem cell and to analyze its characteristics.

METHODSBased on an expression sequence tags(EST) CF948547 which expressed specifically in human embryonic stem cell, the full-length cDNA sequence of a novel gene was cloned by using bioinformatic and molecular biological technique. Its expression profile was analyzed by reverse transcription-polymerase chain reaction(RT-PCR), and subcellular location was determined by enhanced green fluorescent protein (EGFP) eukaryotic expression system.

RESULTSA novel gene HPESCRG1(homo sapiens pluripotent embryonic stem cell-related gene) was cloned successfully. Its GenBank accession number was AY283672. Its cDNA length was 1395 bp. It comprised 9 exons and 8 introns, and its opening reading frame was 250-1146 bp. Its chromosomal mapping was located in 3q13.13, and the putative protein contained 297 amino acids. The theoretical molecular weight of the putative protein was 33 784 and the isoelectric point was 9.35. The protein primary structure of this gene contained a SAP motif and it was subcellularly located in nuclei. Expression analysis showed that this gene was expressed specifically in human ES cells, but not expressed in the adult human tissues, the multiple tissues of embryo aborted in over 5 months' pregnancy, the differentiated cells of HESC-1, and the human mesenchymal stem cells (hMSCs) and human embryonic fibrocytes (hEFCs).

CONCLUSIONHPESCRG1 was found to be a novel gene expressed specifically in human ES cell, which might be related to self-renewal of human ES cell and maintaining its undifferentiated state.

Amino Acid Sequence ; Base Sequence ; Cell Line ; Cloning, Molecular ; DNA, Complementary ; analysis ; Embryo, Mammalian ; Exons ; Expressed Sequence Tags ; Gene Expression ; Humans ; Introns ; Molecular Sequence Data ; Molecular Weight ; Nuclear Proteins ; Open Reading Frames ; genetics ; Proteins ; genetics ; metabolism ; Stem Cells ; cytology ; metabolism

OBJECTIVE: To determine the carrier rate for 21 inherited metabolic diseases among a Chinese population of childbearing age. METHODS: A total of 897 unrelated healthy individuals (including 143 couples) were recruited, and DNA was extracted from their peripheral blood samples. Whole exome sequencing (WES) was carried out to screen potential variants among 54 genes associated with 21 inherited metabolic diseases. Pathogenic and likely pathogenic variants and unreported loss-of-function variants were analyzed. RESULTS: One hundred fourty types of pathogenic/likely pathogenic variants (with an overall number of 183) and unreported loss-of-function variants were detected, which yield a frequency of 0.20 per capita. A husband and wife were both found to carry pathogenic variants of the SLC25A13 gene and have given birth to a healthy baby with the aid of preimplantation genetic diagnosis. The detected variants have involved 40 genes, with the most common ones including ATP7B, SLC25A13, PAH, CBS and MMACHC. Based on the Hardy-Weinberg equilibrium, the incidence of the 21 inherited metabolic diseases in the population was approximately 1/1100, with the five diseases with higher incidence including citrullinemia, methylmalonic acidemia, Wilson disease, glycogen storage disease, and phenylketonuria. CONCLUSION This study has preliminarily determined the carrier rate and incidence of 21 inherited metabolic diseases among a Chinese population of childbearing age, which has provided valuable information for the design of neonatal screening program for inherited metabolic diseases. Pre-conception carrier screening can provide an important measure for the prevention of transmission of Mendelian disorders in the population.
Asians/genetics* ; China ; Exome ; Female ; Humans ; Infant, Newborn ; Metabolic Diseases/genetics* ; Mitochondrial Membrane Transport Proteins/genetics* ; Oxidoreductases/genetics* ; Whole Exome Sequencing

BALB/c mice were immunized with purified White spot syndrome virus (WSSV).Six monoclonal antibody cell lines were selected by ELISA with VP28 protein expressed in E.coll in vitro neutralization experiments showed that 4 of them could inhibit the virus infection in crayfish.Westernblot suggested that all these monoclonal antibodies were against the conformational structure of VP28.The monoclonal antibody 7B4 was labeled with colloidal gold particles and used to locate the VP28 on virus envelope by immunogold labeling.These monoclonal antibodies could be used to develop immunological diagnosis methods for WSSV infection.