Objective To construct the yeast expression vector of a new gene transactivated by hepatitis B virus X protein (XTP11), and to explore the feasibility of cloning the hepatocyte proteins interacting with XTP11 protein by yeast two hybrid system. Methods The XTP11 gene was amplified by polymerase chain reaction (PCR) with specific primers, and the amplified fragment was subcloned into the Nco I/BamH I sites (5′ ends) of yeast expression vector pGBKT7,which was named as pGBKT7(-)-XTP11 encoding fusion proteins of full length of XTP11 and DNA binding domain of yeast protein GAL4. The pGBKT7 (-)-XTP11 plasmid was transformed in the yeast cells AH109 and the expression of XTP11 protein in yeast cells was detected by Western blotting assay. Then the yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp and SD/-Trp-His-Ade) containing X-?-gal, and their autonomous activation was tested. Results The yeast expression vector of XTP11 gene was constructed and the fusion protein in yeast cells was examined. The yeast cells transformed with pGBKT7-XTP11 plasmid could grew well on both of the media, and turned blue on medium containing X-?-gal for the production X-?-galactosidase. This phenomenon suggested that XTP11 protein acted to substitute the functional yeast protein GAL4 activation domain (AD) to activate transcription of reporter genes (ADE2, HIS3, MEL1 and LacZ). Conclusion The XTP11 protein fused to the GAL4 DNA-binding domain functioned as a transcriptional activation domain in yeast, and the transcriptional activation function of XTP11 protein in yeast might restrain researchers to use yeast two hybrid system to clone hepatocyte proteins interacting with XTP11 protein.

OBJECTIVE:To investigate the use of prophylactic use of antibiotics for special diagnosis and treatment in our hos-pital,and to provide reference for rational use of antibiotics. METHODS:The patients receiving special diagnosis and treatment were collected from our hospital during Jan.-Aug. 2016. The prophylactic application of antibiotics was analyzed statistically. RE-SULTS:Among 149 included patients,there were 62 cases of prophylactic use of antibiotics with utilization rate of 41.61％. Among 62 patients receiving antibiotics,5 cases were given medicine orally and others were via injection. Nine categories 18 types of antibiotics were used,involving 69 times;cephalosporin were used in 49 times(71.01％),among which top 3 types in the list of use sequence were cefodizime,cefotetan and cefazolin;prophylactic use of antibiotics was not found in angiography(including coronary artery) and stent implantation. Utilization rate of antibiotics for other diagnosis and treatment was more than 50％. The prophylactic use of antibiotics was complex in ureteroscopy and cystoscopy,shock wave lithotripsy;the selection of drug types started from the high level. There were 59 cases of single drug(95.16％)and 3 cases of two-drug(4.84％). Total of 41 cases were given medicine 0.5-2 h before surgery or at the beginning of anaesthesia (66.13％). Medication time of 18 cases was ≤24 h (29.03％);that of 11 cases was >24-48 h(17.74％). CONCLUSIONS:The prophylactic use of antibiotics is relatively standard in angiography(including coronary artery),stent implantation and implantation of peritoneal dialysis tube in our hospital. There are some problems during special diagnosis and treatment,such as the indication of drug delivery is not strict;the selection of drug types starts from the high level;medication timing is not suitable;medication course is excessively long.

Objective To observe the effect of Yiqi Fuzhi Granule on activating blood circulation and removing blood stasis in rat models of multiple infarctional dementia.Methods Internal carotid artery injection method was used to establish rat models of multiple infarctional dementia.The changes of blood rheology,blood platelet aggregation,thromboplastin time,plasmozyme time and kaolin partial thromboplastin time before and after operation were observed.Results Yiqi Fuzhi Granule could markedly improve blood rheology in rat models of multiple infarctional dementia and reduce blood viscosity and blood platelet aggregati on rate,the effect being obvious before operation.Conclusion Yiqi Fuzhi Granule has a good action of activating blood circulation and removing blood stasis.

OBJECTIVE To study effective and convenient method for paraffin oil sterilization.METHODS By using carrier qualitative germicidal test,to compare pressure steam sterilization,dry heat sterilization and cobalt-60(gamma)-ray radiation sterilization to test the sterilizing effect and operating procedure.RESULTS Pressure steam sterilization was unable to achieve 100% sterilizing effect,whether we extended the time or use the intermittent(sterilization).After dry heat or radiation sterilization processes,no microorganism was found.CONCLUSIONS Effect of sterilization with dry heat or radiation sterilization is trustable,but its packing,operation and equipment are requested strictly,and pressure steam sterilization may be not good for paraffin oil.

Objective:To observe the efficacy of polyene phosphatidyl choline combined with Hugan tablets in the treatment of drug-induced liver injury. Methods:Totally 80 cases of drug-induced liver injury were randomly divided into the observation group (40 cases )and the control group(40 cases). The observation group was given polyene phosphatidyl choline injections 697. 5mg+5%glucose injections 250ml,ivd,qd,combined with Hugan tablets 1. 44g,po,tid,the control group was given polyene phosphatidyl choline injections 697. 5mg+5%glucose injections 250ml,ivd,qd,and the symptomatic treatment in the two groups was the same. The two groups were treated for 20d. Alanine aminotransferase ( ALT), aspartate aminotransferase ( AST), total bilirubin ( TBIL), glutamyl transpeptidase(GGT) and alkaline phosphatase(ALP)were detected respectively before and after the treatment, and the therapeutic effects were compared between the two groups. Results: After the treatment,the total effective rate in the observation group was 97. 5%,which was significantly higher than that in the control group(P <0. 01),Symptoms,signs and biochemical indices in the two groups were improved obviously. The decrease of ALT,AST and GGT in the observation group after the treatment was obviously lower than that in the control group(P<0. 05). Conclusion: Polyene phosphatidylcholine injections combined with Hugan tablets in the treatment of liver injury is better than polyene phosphatidyl choline, which is worthy of clinical application.

Objective To investigate Th1/Th2 pattern of response in psoriasis and to study the effects of CTLA4Ig on Th1/Th2 shift in psoriasis. Methods The levels of IL-2, IFN-?and IL-4 were assessed by ELISA in the supernatant of cultured peripheral blood mononuclear cells (PBMCs) from 33 patients with psoriasis vulgaris and 20 healthy blood donors. Cytokine synthesis was induced by activation with phytohemagglutinin (PHA) and staphylococcal enterotoxin B (SEB) with or without CTLA4Ig. Results Levels of IL-2, IFN-?and IL-4 were markedly increased by PBMCs from psoriatic patients incubated with SEB (P

Objective To investigate the status of expression and gene polymorphism of cytotoxic T lymphocyte-associated molecule4(CTLA4)on peripheral blood mononuclear cells(PBMCs)from patients with psoriasis.Methods Expression of CTLA4mRNA/antigen and polymorphism of CTLA4gene were analyzed in33and133patients with psoriasis confirmed clinically and/or pathologically,respectively.Expression of CT-LA4mRNA and antigen was detected by in situ hybridization and immmunohistochemistry.CTLA4exon1and3'untranslated region were amplified by polymerase chain reaction(PCR)and the amplified products were identified by single-stand conformation polymorphism(SSCP),restriction fragment length polymorphism(RFLP)or sequencing.Results Expression of both mRNA and antigen of CTLA4was significantly weaker on PBMCs induced by staphytococcal enterotoxin B(SEB)in patients with psoriasis than that in normal con-trols,without a regular pattern.The guanine49on exon1in association with fragment106bp of3'untranslat-ed region was shown to be linked to the susceptibility of psoriasis.Conclusion Defective translation and ex-pression of CTLA4take place in patients with psoriasis,which is possibly related to the polymorphism of CT-LA4.Our results suggest that CTLA4may be one of the candidate genes which cause autoimmunity in psoria-sis.

Objective To observe effects of problem-based learning(PBL) combined with case-based study (CBS)teaching mode in clinical teaching in intensive care unit(ICU). Methods Students from ICU of Fangcun Branch of Guangdong Provincial Hospital of Traditional Chinese Medicine from March 2011 to February 2012 were divided into treatment group (31 cases, with PBL combined with CBS teaching mode) and control group (34 cases, with the traditional teaching mode). Scores of two groups were compared and analyzed and questionnaire survey was conducted in treatment group. Data were processed by SPSS 13.0 statistical software. Measurement data were compared by t-test and data of heterogeneous variance were compared by rank-sum test. Results Treatment group achieved better scores ((85.26 ±5.96) in theory examination and (80.59 ±7.33) in case-analyzing examination) compared with those of control group ((79.17 ±7.31) in theory examination and (76.02 ±9.27) in case-analyzing examination)(P<0.05). PBL combined with CBS teaching mode can stimulate learning interests but it was lack of systematization; most students did not volunteer to speak. Conclusion PBL combined with CBS teaching mode has certain advantages in ICU clinical teaching.

Objective By detecting the expression levels of Foxp3 in CD4+CD25+ regulatory T cells of peripheral blood mononuclear cells (PBMCs) in rheumatoid arthritis (RA),and the Foxp3 gene promoter region methylation to explore its role in the pathogenesis of RA.Methods Twenty-five RA patients and 10 healthy controls were selected,and the PBMCs were extracted by density gradient centrifugation.Foxp3 expression levels of CD4+CD25+ regulatory T cells were detected by flow cytometry.The real-time fluorescence quantitative PCR assay was used to detect the Foxp3 mRNA expression in PBMCs; and bisulfate processing gene sequencing was used to determinethe differences in Foxp3 gene promoter sequence methylation level of PBMCs.The comparison between groups was analyzed using one-way ANOVA; two sets of qualitative data were compared using Fisher's exact test.Results The expression levels of Foxp3 mRNA in the CD4+CD25+regulatory T cells of active RA patients (2.31±0.25) was significantly lower than inactive RA group (3.68±0.26) and healthy controls (5.67±0.34),the difference was statistically significant (P＜0.05).The Foxp3 mRNA expression level in inactive RA group was lower than that of the healthy controls (P＜0.05).Foxp3 promoter region-67,-74 sites of methylation level in PBMCs of RA patients (46％) was significantly higher than that of the healthy controls (6％).Conclusion Reduction in the number of CD4+CD25+ regulatory T cells may be involved in the pathogenesis of RA and Foxp3 gene promoter methylation levels plays a key role in this process.

OBJECTIVETo determine the mutational profile and clinical implications of the viral reverse-transcriptase (rt)A 181T mutation in hepatitis B virus (HBV) through population-based analysis of clinical samples.

METHODSSerum samples from 3, 013 patients who visited The 302 Hospital (Beijing, China) were investigated.HBV DNA was extracted and HBV mutations and genotypes were determined by direct sequencing.Recombinant plasmids harboring the rtA181T/sW172* mutant or wild type sequence were constructed and transfected into the HepG2 cell line. The levels of HBsAg in culture supernatants were compared and statistically analyzed.

RESULTSThe incidence of rtA181T across the study population was 4.1% (165/3, 013), and most of the rtAl 81T-positive patients had received adefovir and/or lamivudine.Forty percent (66/165) of the rtA 181T cases were single mutants and treatment responsive, 46.1% (76/165) included the adefovir-resistant mutation rtA 181 V/N236T, 12.1% (20/165) included the lamivudine-resistant mutation rtM204V/rtM2041, and 1.8% (3/165) included multidrug-resistant mutations.Interestingly, 73.9% (122/165) of the rtA181T-positive samples were detected with co-existing wild-type nucleotides at the site. The rates of HBV/C to HBV/B were 92.1% to 7.9% in the rtA181T-positive patients, but 82.1% to 17.9% in the rtA181T-negative paticnts (P less than 0.01).Almost all (98.2%; 129/165) of the rtA181T led to sW172*, while only 1.8% of the rtA181T (3/165) led to sW172L or sW172S.HBsAg secretion in vitro was reduced from the rtA181T/ sW172* strain, but there was no significant difference observed in the average serum HBsAg and HBV DNA levels of patients who carried or did not carry the mutant.

CONCLUSIONThe HBV rtA181T mutation is closely associated with adefovir and lamivudine exposure.rtA181T may led to sW172*, culminating in suppression of HBsAg secretion.However, co-existence of the mutant with wild-type sequences was common among our patient population, suggesting that the mutation had little impact on serum HBsAg and HBV DNA levels across the clinical study population.

Adenine ; analogs & derivatives ; Antiviral Agents ; China ; Genotype ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Humans ; Lamivudine ; Mutation ; Organophosphonates