Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Hepatitis B ; epidemiology ; Humans ; Infant ; Middle Aged ; Risk Factors ; Seroepidemiologic Studies ; Young Adult

Objective To study the exact function of human gene 5 transactivated by pre-S1 protein of hepatitis B virus(PS1TP5)by investigating the gene expression of PS1TP5 in yeast cells. Methods Reverse transcription-polymerase chain reaction(RT-PCR)was performed to amplify the gene of PS1TP5 using the mRNA of HepG2 cells as template and the gene was cloned into pGEM-T vector.The gene of PS1TP5 was cut from pGEM-T-PS1TP5 vector and cloned into yeast expressive plasmid pGBKT7,then pGBKT7-PS1TP5 was transformed into yeast cell AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) and Western hybridization.Results PS1TP5 gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The results of SDS-PAGE and Western assay showed that the relative molecular weight of the expressed product was about 36 950,and PS1TP5 protein existed in yeast cells.Conclusion The findings suggest that PS1TP5 can be successfully expressed in yeast cell.

OBJECTIVETo evaluate the incidence of postoperative hemorrhage in children undergoing adenoidectomy, and to discuss its possible causes.

METHODSIncluded in this study were children who underwent adenoid and/or tonsil surgery at Shenzhen Children's Hospital between January 2004 and November 2009. The change of hemoglobin (Hb) and hematocrit (Hct) were retrospectively analysed. The blood loss was estimated by the change of Hct.

RESULTSThere were 2078 cases that accomplished the inclusion criteria in the period of study. Ten children bled 0.5 - 4.0 hours after surgery, without superfluous hemorrhage during the operation and post-tonsillectomy. This represented an incidence of 0.48%of immediate postoperative haemorrhage among the 2078 procedures analyzed. Statistical differences were found between boys (0.21%) and girls (1.10%, χ² = 5.597, P < 0.05). The change of Hb and Hct was positively correlated (r = 0.95, P < 0.01), the blood loss was positively correlated with the bleeding time (r = 0.66, P < 0.05). The causes of postoperative hemorrhage were coagulation system deficits, chronic nasopharyngitis, deficient hemostasis and immoderate ravage. To control the postoperative hemorrhage, 2 postnasal packing under topical anaesthesia and 8 electrocautery under general anaesthesia were applied.

CONCLUSIONSPoor operative technique and deficient hemostasis are the major causes of primary hemorrhage. Prompt operation to control the postoperative bleeding should be done 2 hours after bleeding under general anesthesia in order to avoid severe complications.

Adenoidectomy ; adverse effects ; Adolescent ; Child ; Child, Preschool ; Female ; Hematocrit ; Hemoglobins ; analysis ; Humans ; Infant ; Male ; Postoperative Hemorrhage ; etiology ; Retrospective Studies ; Tonsillectomy ; adverse effects

OBJECTIVETo investigate the health and work status of traffic policemen in a city, and analyze the effect of the occupation on health of them.

METHODSThe prevalent investigation was used to investigate the health status of on-the-job traffic policemen and the effect of the occupation on the health of traffic policemen was studied.

RESULTSAmong 4486 traffic policemen, 1604 working outside. The average time of working outside was (12.0 +/- 7.9) years and the average time of standing was (7.0 +/- 1.7) hours/day. The incidence of naso-pharyngitis, arthropathy, photosensitive dermatitis, heat stroke and chilblain, noise-induced deafness in the outside-traffic policemen were higher than that of the policemen working indoors. Eye diseases were higher in those who worked outside for 20-25 years. The varicosis of the lower extremity was higher in those who stood outside over 4 hours every day. The average period of onset of disease was 7-8 years.

CONCLUSIONThe occupational stress of outside-traffic policemen was bigger in the city. The environmental pollution and the bad condition are the risky factors of 8 kinds of diseases among traffic policemen. The average period of onset of the disease is 7-8 years.

Adult ; China ; Female ; Health Status ; Humans ; Male ; Middle Aged ; Occupational Diseases ; epidemiology ; Occupational Exposure ; Occupational Health ; Police ; Urban Population ; Young Adult

BACKGROUNDThe hepatitis B virus (HBV) genome includes S, C, P and X regions. The S region is divided into four subregions of pre-pre-S, pre-S1, pre-S2 and S. PS1TP5 (human gene 5 transactivated by pre-S1 protein of HBV) is a novel target gene transactivated by the pre-S1 protein that has been screened with a suppression subtractive hybridization technique in our laboratory (GenBank accession: AY427953). In order to investigate the biological function of the PS1TP5 protein, we performed a yeast two-hybrid system 3 to screen proteins from a human leukocyte cDNA library interacting with the PS1TP5 protein.

METHODSThe reverse transcription polymerase chain reaction (RT-PCR) was performed to amplify the gene of PS1TP5 from the mRNA of HepG2 cells and the gene was then cloned into the pGEM-T vector. After being sequenced and analyzed with Vector NTI 9.1 and NCBI BLAST software, the target gene of PS1TP5 was cut from the pGEM-T vector and cloned into a yeast expression plasmid pGBKT7, then "bait" plasmid pGBKT7-PS1TP5 was transformed into the yeast strain AH109. The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting hybridization. After expression of the pGBKT7-PS1TP5 fusion protein in the AH109 yeast strain was accomplished, a yeast two-hybrid screening was performed by mating AH109 with Y187 containing a leukocyte cDNA library plasmid. The mated yeast was plated on quadruple dropout medium and assayed for alpha-gal activity. The interaction between the PS1TP5 protein and the proteins obtained from positive colonies was further confirmed by repeating the yeast two-hybrid screen. After extracting and sequencing of plasmids from blue colonies we carried out a bioinformatic analysis.

RESULTSForty true positive colonies were selected and sequenced, full length sequences were obtained and we searched for homologous DNA sequences from GenBank. Among the 40 positive colonies, 23 coding genes with known functions were obtained, including Homo sapien leukocyte adhesion protein p150, 95, interleukin 2 receptor gamma chain, PALM2-AKAP2 protein (PALM2-AKAP2), eukaryotic translation initiation factor 4A, beta-2-microglobin, solute carrier family 9 (sodium/hydrogen exchanger), calreticulin, asialoglycoprotein receptor 1 (ASGR1), MHC class II lymphocyte antigen, cytochrome c oxidase subunit 1, lymphocyte antigen 86 (LY86) and lymphocyte cytosolic protein 1. One novel gene with unknown function was found and named as PS1TP5BP1. After being electronically spliced, it was deposited in GenBank (accession number: DQ471327).

CONCLUSIONSGenes of proteins interacting with PS1TP5 were successfully screened from leukocyte cDNA library. These results suggested that PS1TP5 was closely correlated with immunoregulation, carbohydrate metabolism, signal transduction, the formation of hepatic fibrosis and initiation and development of tumors and also brought some new clues for further studying the biological functions of the pre-S1 protein.

Amino Acid Sequence ; Base Sequence ; DNA, Complementary ; chemistry ; Gene Library ; Hepatitis B Surface Antigens ; genetics ; physiology ; Humans ; Leukocytes ; metabolism ; Molecular Sequence Data ; Plasmids ; Protein Interaction Mapping ; Protein Precursors ; genetics ; physiology ; Recombinant Fusion Proteins ; biosynthesis ; Transcriptional Activation ; Two-Hybrid System Techniques ; Yeasts ; genetics

Abstract: Our previous work revealed berberine can significantly enhance the susceptibility of fluconazole against fluconazole-resistant Candida albicans, which suggested that berberine has synergistic antifungal activity with fluconazole. Preliminary SAR of berberine needs to be studied for the possibility of investigating its target and SAR, improving its drug-likeness, and exploring new scaffold. In this work, 13-substitutited benzyl berberine derivatives and N-benzyl isoquinoline analogues were synthesized and characterized by 1H NMR and MS. Their synergetic activity with fluconazole against fluconazole-resistant Candida albicans was evaluated in vitro. The 13-substitutited benzyl berberine derivatives 1a-1e exhibited comparable activity to berberine, which suggested that the introduction of functional groups to C-13 can maintain its activity. The N-benzyl isoquinolines, which were designed as analogues of berberine with its D ring opened, exhibited lower activity than berberine. However, compound 2b, 2c, and 4b showed moderate activity, which indicated that berberine may be deconstructed to new scaffold with synergistic antifungal activity with fluconazole. The results of our research may be helpful to the SAR studies on its other biological activities.
Antifungal Agents ; pharmacology ; Berberine ; pharmacology ; Candida albicans ; drug effects ; Drug Resistance, Fungal ; Drug Synergism ; Fluconazole ; pharmacology ; Isoquinolines ; pharmacology ; Microbial Sensitivity Tests

OBJECTIVETo evaluate the effects of prevention and control strategies on hepatitis A.

METHODSSurveillance data on hepatitis A from 1990 to 2006 in Tianjin was analyzed, and the coverage rate of hepatitis A vaccine among targeted population was estimated, to compare the anti-HAV IgG level of children younger than 15 years old in 1999 and in 2005.

RESULTSResults showed that a) the morbidity of hepatitis A decreased from 25.26/10(5) in 1990 to 0.82/10(5) in 2006; b) the ratio of hepatitis A in viral hepatitis decreased from 30.43% in 1990 to 1.05% in 2006; c) the estimated coverage rate was 72.7%; d) the positive rate of anti-HAV among children younger than 15 years old in 2005 was distinctly higher than that in 1999.

CONCLUSIONPositive results showed that it was successful to use hepatitis A vaccine as the strategy to prevent and control hepatitis A in the past five years in Tianjin.

Adolescent ; Child ; Child, Preschool ; China ; epidemiology ; Hepatitis A ; epidemiology ; prevention & control ; Hepatitis A Vaccines ; administration & dosage ; Hepatitis Antibodies ; blood ; Humans ; Immunization Programs ; Incidence ; Infant ; Population Surveillance

OBJECTIVETo analyze tracheal intubation and respiratory treatment in the critical severe acute respiratory syndrome (SARS) patients.

METHODSReview and analyze tracheal intubation and respiratory treatment in critical SARS patients in intensive care unit (ICU).

RESULTSThree of thirteen patients had been intubated or received tracheotomy before they entered into ICU, the other patients received treatment of nasal cannula or oxygen mask. With the development of the disease, two patients had been intubated because of respiratory failure or tracheotomy. Tracheal intubation was twice made in two patients in order to replace tracheal tubes.

CONCLUSIONSThe patient should be intubated or received tracheotomy if non-invasive respiratory support has no effect. Standard protection could protect medical staff from infection under tracheal intubation.

Adult ; Aged ; Aged, 80 and over ; Critical Care ; Evaluation Studies as Topic ; Female ; Humans ; Infectious Disease Transmission, Patient-to-Professional ; prevention & control ; Intubation, Intratracheal ; methods ; Male ; Middle Aged ; Severe Acute Respiratory Syndrome ; therapy ; transmission ; Tracheostomy

OBJECTIVETo investigate the biological functions of TTG1A in liver fibrosis.

METHODSYeast two-hybrid system was used to screen proteins associated with TTG1A. Briefly, the coding sequence of TTG1A was cloned into pGBKT7 vector, and the recombinant plasmid was transformed into yeast cells AH109 ( a type), then these cells were mated with yeast cells Y187 (a type) transformed with human leukocyte cDNA library plasmid pACT2. The obtained diploid yeast cells were plated on synthetic dropout nutrient medium containing X-alpha-gal for double selection. The plasmids from positive colonies were transformed into E.coli and sequenced.

RESULTSThe recombinant yeast expression vector pGBKT7-TTG1A was successfully constructed. Nineteen TTG1A binding proteins, including Homo sapiens major histocompatibility complex, class II DP beta 1 (HLA-DPb1), Homo sapiens ribosomal protein L30 (RPL30), Homo sapiens nucleophosmin Homo sapiens nucleobindin 2 (NUCB2), Homo sapiens ash2, variant Gaucher disease and variant metachromatic leukodystrophy, MORF4L1, Homo sapiens ubiquitin-conjugating enzyme E2L3 (UBE2L3), APOA1, Homo sapiens lectin, and galectin 1, were identified.

CONCLUSIONSThis study may help to elucidate the molecular function of TTG1A.

Carrier Proteins ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Library ; Genes, Regulator ; Genetic Vectors ; Hepatic Stellate Cells ; Humans ; Liver Cirrhosis ; genetics ; Oligonucleotide Array Sequence Analysis ; Plasmids ; genetics ; Ribosomal Proteins ; genetics ; Transcriptional Activation ; Transforming Growth Factor beta1 ; genetics ; Two-Hybrid System Techniques ; Yeasts ; genetics

OBJECTIVETo study the inhibitory effect of T-2 toxin on the expression of aggrecan and collagen II in chondrocytes and the protection of selenium against this effect.

METHODSHuman chondrocytes cultured in vitro were treated with T-2 toxin at different concentrations for varied time periods (1-5 days), and the cell viability was measured by MTT assay. Aggrecan expression was detected by toluidine blue staining and collagen II expression by immunostaining using monoclonal antibody of collagen. Aggrecan and collagen II mRNA expressions were measured by semiquantitative RT-PCR.

RESULTST-2 toxin dose- and time-dependently affected chondrocyte viability within the concentration range of 0.001-2 mg/L, the prolonged treatment time further enhanced the dose dependence of the inhibitory effect. T-2 toxin lowered aggrecan and collagen II synthesis in the chondrocytes and reduced their mRNA expressions. Selenium could partly attenuate the inhibitory effects of T-2 toxin on aggrecan mRNA expression, but showed no such effect against T-2-induced collagen II expression.

CONCLUSIONT-2 toxin can obviously inhibit aggrecan and collagen II synthesis in human chondrocytes, and selenium can partly antagonize the inhibitory effects of T-2 toxin on aggrecan.

Aggrecans ; biosynthesis ; genetics ; Cells, Cultured ; Chondrocytes ; cytology ; metabolism ; Collagen Type II ; biosynthesis ; genetics ; Dose-Response Relationship, Drug ; Fetus ; Humans ; Protective Agents ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Selenium ; pharmacology ; T-2 Toxin ; toxicity