Objective To investigate the activity of HBV pre-S2 protein on thioredoxin reductase 1 (TXNRD1) gene promoter. Methods TXNRD1 gene promoter DNA sequence was identified in GenBank by bioinformatics and amplified from HepG2 genome by polymerase chain reaction (PCR). The amplified product was cloned into pCAT3-Basic reporter vector,named pCAT3-TXNRD1p. The HepG2 cells were transfected by pCAT3-TXNRD1p,and then co-tranfected by pCAT3-TXNRD1p and pcDNA3.1(-)-preS2 plasmids. The choloraphenical acetyltransferase(CAT)activity was assessed by enzyme linked immunosorbent assay(ELISA). Results The results indicate that HepG2 cells transfected by pCAT3-TXNRD1p had higher activity of CAT than that transfected by pCAT3-Basic. The expression of CAT in HepG2 cells co-transfected by pCAT3-TXNRD1p and pcDNA3.1(-)-preS2 was 2.2 times higher than that with pCAT3-TXNRD1p. Conclusions The TXNRD1 gene promoter identified in this study has transcription activity and HBV pre-S2 protein can transactivate the expression of TXNRD1 gene.

Objective To screen promoter binding protein of cyclin B2 by using human liver cDNA library, and investigate the expression and regulation mechanism of cyclin B2 gene. Methods By using cyclin B2 biotinylated promoter DNA as the selective molecule, the T7select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment, and the DNA fragment was cloned into pGEM-Teasy vector. Results Sequence analysis was performed in 20 positive plaques, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 6 coding sequences were obtained, all of which were known ones. Conclusion Cyclin B2 promoter binding proteins were screened. The results will be useful for further study the expression and regulation mechanism of cyclin B2.

Objective To clone and identify human genes transactivated by homo sapiens HCV FTP2 by constructing a cDNA subtractive library with suppression subtractive hybridization tech- nique.Methods Suppression subtractive hybridization(SSH)and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV FTP2.The mRNA was iso- lated from HepG2 cells transfected pcDNA3.1(-)-HCV FTP2 and pcDNA3.1(-)empty vector re- spectively,and SSH method was employed to analyze the differentially expressed DNA sequence be- tween the two groups.After digestion with restriction enzyme Rsa I,small-size cDNAs were ob- tained.Then tester cDNA was divided into two groups and ligated to the specific adaptor I and adap- tor 2 respectively.After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR and then was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain DH5?.Futhermore,the cDNA was se- quenced and analyzed in GenBank with Blast search after PCR.Results The subtractive library of genes transactivated by HCV FTP2 was constructed successfully.The amplified library contains 71 positive clones.Colony PCR shows that 56 clones contain 200～1000 hp inserts.Sequence analysis was performed in 24 clones randomly,and the full length sequences were obtained with bioinformatics method.Altogether 20 coding sequences in total were obtained,consisting of 19 known and 1 un- known.Conclusion The obtained sequences may be target genes transactivated by HCV FTP2,and some genes coding proteins involved in cell cycle regulation,metabolism and cell apoptosis.

Objective To study the exact function of HBeBP4A so as to investigate the gene expression of HBeBP4A in yeast cell.Methods Reverse transcription polymerase chain reaction(RT-PCR)was employed to amplify the gene of HBeBP4A from recombinant plasmids pcDNA 3.1/myc-HisA-HBeBP4A,and the gene was cloned into pGEM-T vector.The gene of HBeBP4A was cut from pGEM-T-HBeBP4A vector and cloned into yeast expressive plasmid pGBKT7,and pGBKT7-HBeBP4A was then transformed into yeast AH109.The yeast protein was isolated and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and Western blotting hybridization.Results HBeBP4A gene was successfully amplified and identified by DNA sequencing.The digested fragment was cloned into pGBKT7 vector and transformed into yeast cell AH109.The SDS-PAGE and Western blotting assay showed that the relative molecular weight of the expressed product was about 61.37kD,and HBeBP4A protein existed in yeast cells.Conclusion The findings suggested that HBeBP4A was successfully expressed into yeast system.

Objective To clone a new human gene 5 trans-activated by pre-S1 protein of hepatitis B virus (HBV), PS1TP5, and explore its structure and function by bioinformatics analysis. Methods PS1TP5 was amplified by reverse transcription-polymerase chain reaction (RT-PCR) technique by using HepG2 cDNA as template and inserted into pGEM-T vector by TA cloning. Recombinant eukaryotic expression vector pcDNA TM 3.1/myc-His A-PS1TP5 had been constructed by subcloning, followed by restriction enzyme digestion analysis and sequencing. Bioinformatic methods were used to analyze its possible physical and chemical characters, structure, and function. Results PS1TP5 was successfully amplified and cloned into pGEM-T and pcDNA TM 3.1/myc-His A vector by RT-PCR from HepG2 cDNA. The new gene had been confirmed by sequencing after PCR identification and restriction enzyme digestion and named as PS1TP5 because of its trans-active function. The sequence for the PS1TP5 gene had been deposited into GenBank, the accession number was AY427953. Bioinformatics analysis showed that its ORF was 438bp and translated a protein of 145 aa. Conclusion A new gene-PS1TP5 has been recognized, and its recombinant eukaryotic expression vector (pcDNA TM 3.1/myc-His A-PS1TP5) has been constructed. These results will certainly bring some new clues for the study of the biological function of new gene and pathogenesis of chronic hepatitis B.

Objective To evaluate the relationship of central aortic pressure (CAP) with atherosclerosis and left ventricular function in elderly patients with essential hypertension. Methods A total of 155 elderly hypertensive patients were divided into two groups: aged 60-79 years group (n = 71) and aged 80-95 years group (n= 84). Central aortic waveforms were generated using pulse wave analysis, then CAP and augmentation index (AI) were determined. Auto-survey atherosclerosis apparatus was applied to examine brachial-ankle pulse wave velocity (baPWV), ankle-brachial index (ABI) and toe-brachial index (TBI). Interventricular septal thickness at end diastole (IVSd), left ventricular end-diastolic dimension (LVDd), left ventricular posterior wall thickness at end diastole (LVPWd), relative left ventricle thickness (RLVT), left ventricular mass index (LVMI), Ejection fraction(EF) slope, left ventricle ejection fraction (LVEF) and fractional shortening (FS) were measured by the two-dimensional echoeardiography. Results Systolic pressure (SBP), pulse pressure (PP), CAP, AI and baPWV were significantly higher in aged 80-95 years group than in aged 60-79 years group (all P<0.05), ABI and TBI were significantly lower oppositely (both P<0. 01). IVSd, LVPWd, RLVT and LVMI were all significantly higher and EF slope was lower in aged 80-95 years group than in aged 60-79 years group (all P<0. 057. There were no significant differences in LVDd, LVEF and FS between the two groups (both P>0. 05). CAP had positive association with PP, AI and baPWV (r=0. 505,0. 284,all P<0.01). After adjustment for age, gender, smoking, body mass index, fasting blood sugar, creatinine, uric acid, cholesterol, triglyceride, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol, there was no significant relationship between CAP and ABI or TBI (both P>0. 05). There was also positive association of CAP with IVSd, LVPWd, RLVT, LVMI, while negative associations of CAP with EF slope (all P<0. 01). There were no significant relationship between CAP and LVEF, FS, LVDd (all P> 0.05). Conclusions CAP and degree of artherosclerosis increase with aging in elderly patients with essential hypertension, which contributes to left ventricular hypertrophy and the decreased diastolic function. CAP helps to make an early diagnosis of or screening arteriosclerosis, and it is an important forecast factor for cardiovascular disease.

OBJECTIVE:To establish a HPLC method for determining the contents of rhein and emodin in Jianfei Jiangzhi tablets.METHODS:Stationary phase was Waters Symmetry C 18 (250mm?4.6mm;5?m),mobile phase was menthol-0.01%H 3 PO 4 (90∶10),detection wavelength440nm,flow rate1.0ml/min and column temperature30℃.RESULTS:There was good linearity in range of0.108?g～0.468?g for rhein and in range of0.180?g～0.480?g for emodin.The average recoveries of rhein and emodin were99.7%and98.6%respectively;RSD were1.93%and2.19%respectively.CONCLUSION:This method is rapid and accurate.It can be applied to effective control of the quality of Jianfei Jiangzhi tablet.

The authors report one case of primary hepatic neuroendocrine carcinoma associated with multiple liver metastases. A patient was a 41-year-old female. In 2010, B-ultrasound examination revealed that there were multiple space-occupying lesions in the liver, and hepatic hemangiomas was considered to be the diagnosis. Then, the patient was followed up regularly. In Aug. 2013, B-ultrasound examination indicated that the hepatic lesions were significantly enlarged. Multi-detector CT scanning and MRI examination were performed, and still the diagnosis of multiple hepatic hemangiomas was suggested. On CT and MRI the lesion presented as a well-circumscribed hypervascular tumor with “fast-in and slow-out” enhancement pattern. On MRI, the lesion was characterized by multiple nodules. Needle biopsy was carried out, and the pathological and immunohistochemical diagnosis was metastatic neuroendocrine tumor. Systemic examination did not find the primary lesion. Therefore, primary hepatic neuroendocrine carcinoma associated with intra- hepatic metastases was diagnosed. The patient was treated with transcatheter arterial chemoembolization. The drugs used were 100 mg Oxaliplatin+one bottle of gelatin sponge particles(300-500μm)+10 ml iodized oil, and micro-pump infusion of 100 mg oxaliplatin(99 mg/h) through catheter was also employed. Clinically, primary hepatic neuroendocrine carcinoma is extremely rare. In combination with the medical literatures, the authors attempt to make a preliminary discussion on the clinical characteristics, differential diagnosis, treatment and prognosis of primary hepatic neuroendocrine carcinoma.

Objective To investigate work stress state of nursing home staff and to analyze the influential factors affecting work stress.Methods Stratified random sampling method was adopted to investigate 180 nursing assistants by self-designed questionnaire,Work Stress Scale(WSS),Simplified Coping Style Questionnaire and internal-external locus of control scale investigation.Results Totally 174 valid questionnaires were collected.Variance analysis revealed there were significant differences of age and other factors.Stepwise multiple regression analysis showed five variables entered the equation,including workexpenence education level number of the cared ages work scheduline coping stule and explain 56.7％ of the total variable.Conclusion Nurse managers should pay attention to and improve achievement sense and mental adjustment ability of nursing assistants,to build a well-organized support system to reduce the workload.

Objective To observe the rate-adaptive response of dual sensor pacemakers integrated with activity sensor and minute ventilation sensor.Methods Fifty patients with chronotrepic incompetence were implanted with pacemakers of integrated sensor.The patients with pacemakers implanted were arranged to take upstairs test,downstairs test and tapping test under the monitor of integrated sensor,minute ventilation sensor and activity sensor respectirely.The rate-adaptive responses were tracked down every 15 seconds with remote heart rate monitor.The results were compared with the control group patients with normal sinal chronotropic function(n=15).Results In activity sensor group,pacing rates were significantly higher than the normal control group in downstairs test and tapping test.In minute ventilation sensor group,pacing rates increasement was significantly slower in the beginning of upstairs test.Pacing rates of integrated group were similar to the normal control group in upstairs and downstairs tests.Conclusion The rate-adaptive pacing response of dual sensor integrated with activity sensor and minute ventilation sensor,which is mostly close to the normal chronotropic response,better than that of single sensor.