OBJECTIVETo investigate the apoptosis regulation on hepatoma cells by HBx between genotype B and C.

METHODSGenotype B and C HBx gene fragments were amplified and inserted into green fluorescent protein (GFP) eukaryotic expression vector pEGFP-C1 to construct recombinant pGFP-XB and pGFP-XC. The pEGFP-C1, pGFP-XB and pGFP-XC were introduced into Bel-7402 cells by Fugene HD to obtain Bel-7402 cells expressing GFP. The transcription and expression of HBx gene were demonstrated by RT-PCR and Western Blot analysis. Bel-7402, Bel-7402/GFP, Bel-7402/GFP-XB, Bel-7402/GFP-XC cells were treated with adriamycin (2.5 microg/ml), and the apoptosis of the cells was determined by trypan blue exclusion, and flow cytometry analysis.

RESULTSRT-PCR and Western Blot analysis showed that HBx genes of genotypes B and C were transcribed and expressed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Trypan blue exclusion showed adriamycin induced time-dependent cell death in Bel-7402, Bel-7402/GFP cells while no significant cell death was observed in Bel-7402/GFP-XB, Bel-7402/GFP-XC cells. Flow cytometry analysis indicated that no significant differences of apoptosis rates of Bel-7402/GFP-XB (3.87%) and of Bel7402/GFP-XC (4.01%) were observed (P > 0.05), moreover, no significant differences of Bel-7402/ GFP-XB (3.87%), Be17402/GFP-XC (4.01%) and of the untreated cells. Apoptosis rates in Bel-7402/GFP-XB (3.87%), Bel-7402/GFP-XC (4.01%) cells were significantly lower than those in Bel-7402 (27.05%) and Bel-7402/GFP (29.14%) cells at 48 hours after the adriamycin treatment (P < 0.01).

CONCLUSIONSBel-7402 cell lines expressing GFP, GFP-XB and GFP-XC fusion proteins were successfully established. HBV X protein blocks adriamycin-induced apoptosis of Bel-7402 cells. There is no difference between HBx of genotype B and C in inhibiting apoptosis induced by adriamycin.

Antiviral Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Hepatitis B ; drug therapy ; physiopathology ; virology ; Hepatitis B virus ; classification ; drug effects ; genetics ; metabolism ; Humans ; Trans-Activators ; genetics ; metabolism

Consensus Development Conferences as Topic ; Endoscopy ; Europe ; Humans ; Nose Neoplasms ; diagnosis ; surgery ; Skull Base Neoplasms ; diagnosis ; surgery

AIM:To determine Radix et Rhizoma Rhici in Ganning Granula(Radix et Rhizoma Rhei,Rhizoma Chuanxiong). METHODS: In order to determine five anthraquinone aglycones of Radix et Rhizoma Rhei,the HPLC system consisted of Nucleosil ODS column,methand-0.1% phosphoric acid(85∶15) mixture as mobile phase,with detection wavelength at 430 nm. RESULTS: HPLC showed that the linearity ranges were as follows: aloe-emodin concentration was at the range of 0.041 6-0.291 2 ?g,the rhein concentration was at the range of 0.037 1-0.259 8 ?g,the emodin concentration was at the range of 0.045 8-0.320 3 ?g,the chrysophanol concentration was at the range of 0.049 60.347 2 ?g,the physcion concentration was at the range of 0.020 6-0.144 5 ?g.The average recoveries of five anthraquinone aglycones were between 96.6%-98.4%.CONCLUSION: The method is simple,accurate,sensible and with good repeatability and can be used for quality control of Ganning Granule.

Objective To explore the reasons why patients with Keshan disesse complicated with hypertension and their interaction in Fuyu County, Heilongjiang Province. Methods Fifty-three patients with Keshan disease were investigated in January, April and July in 2007. Blood pressure was measured and the risk factors of hypertension were investigated. According to the diagnostic criteria of hypertension, patients were divided into hypertension group and non-hypertension group, and then the risk factors of hypertension, as well as the course of Keshan disease, were compared between the two groups. The risk factors include age, gender, family history of hypertension, salt intake in diet, smoking, drinking and obesity. Results The age of hypertension group[(57.83±8.89)years] was significantly higher than that of non-hypertension group [(51.53 ± 9.43)years, t = 2.3630, P < 0.05) ;while the course of Keshan disease in non-hypertension group [(31.63 ± 8.66)years] was notably longer than that in hypertension group [(25.08±11.41)years, t = 2.0224, P < 0.05] ;No statistically significant difference in gender, family history of hypertension, salt intake in diet, smoking, drinking and obesity was observed between the two groups(χ2 = 0.0072,0.1779,0.0029,0.1555,0.119,0.7679, all P > 0.05). Conclusions Age might be an important factor in patients with Keshan disease accompanied by hypertension, and the role of other risk factors of hypertension should not be overlooked;whether Keshan disease and hypertension can affect each other needs further investigation.

Calcaneal fracture is a kind of common injury of foot. Calcaneal fracture could be treated with conservative therapy, open reduction and internal fixation (ORIF), minimal invasive poking reduction and Kirschner wire fixation, arthrodesis or external fixation and other treatment methods. Open reduction and internal fixation as the main treatment method of calcaneus fracture, especially for intra-articular calcaneal fractures, as Sanders type II and III fractures, could gain satisfactory effect. But whether bone grafting should be taken is controversial in Sanders type IV fractures, because soft tissue complications are disadvantages of ORIF. The treatment of minimally invasive percutaneous poking reduction and internal fixation for calcaneal fractures has been paid more and more attention. Because of the differ of the classification of calcaneal fractures, indications of poking reduction and fixation methods, efficacy evaluation standard of choose, the clinical efficacy is different. Because of the complex treatment process, various treatment methods have their own advantages and disadvantages. Through consulting relevant literatures of domestic and abroad in recent years, comprehensive analysis is made and new progress of the calcaneal fractures treatment is summarized.
Calcaneus ; injuries ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Minimally Invasive Surgical Procedures

Adult ; Aged ; Aged, 80 and over ; Coal Mining ; Humans ; Lung Neoplasms ; complications ; microbiology ; Male ; Methicillin Resistance ; Microbial Sensitivity Tests ; Middle Aged ; Pneumoconiosis ; complications ; microbiology ; Staphylococcus aureus ; drug effects ; isolation & purification

The potential of Rhizobium sp. N613 to produce the exopolysaccharide (REPS) was studied in this paper. Using an orthogonal design in a flask-shaker culture system, the fermentation medium and conditions of synthesizing REPS were optimized. Based on these results, the fermentation kinetic parameters were obtained in the batch fermentation with a 10L fermentor. The REPS yield of 11.31g/L was achieved by metabolic regulation during 40 h fed-batch fermentation. Transplanted tumor models of sarcoma 180 in mice were used to evaluate the anti-tumor effect. The result of anti-tumor activities showed that inhibition rate was 53.40%, when dose of REPS was 5mg/kg. These results indicate that REPS has the following properties: the short duration of fermentation, the high yield, the low cost, the effective immunocompetence and thickening. Thus, REPS has the value of development and application.

The potential of a Sinorhizobium fredii strain producing a copolymer polyhydroxyalkanoate(PHA)from glucose and sodium decanoate substrates was studied in this paper.Using orthogonal design in a flask-shaker culture system,the culture medium,some culture conditions and vital regulation conditions for polymer synthesis were optimized.These optimized results were applied into further studies in two-stage fed-batch with a 10L fermentor.The whole culture process consisted of two stages,that is,the cell growth and the copolymer production.The first stage was for the cell growing to a desired biomass and the second was for the copolymer synthesis.For producing PHA polymers,the selected 8 mM sodium decanoate was added into the broth by adopting a two-step adding method for avoiding of foaming when the biomass had approached 28.5g/L dry cell.The maximum P(HB-HH)production could be 17.55 g/L with a monomer ratio of 79.4%(W/W)3-HB and 20.6%(W/W)3-HH.The molecule constitute of the copolymer is poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)[P(HB-HH)] and its molecular weight(MW)is 1.4?105D.The results demonstrated that the employed S.fredii strain could be a potential candidate for industrial production of the copolymer.The fermentation parameters acquired in the experimental system offered some valuable references for studying large-scale production of the copolymer.

Four screening methods, colorimetric assay, blood-plate hemolysis method, surface tension activ- ity and oil spreading technique were introduced to isolate strains producing bio-demulsifiers from 6 different bacteria source samples. The results of various screening methods were evaluated in this paper. Seventeen demulsifying strains were obtained, which are qualified in demulsification test of kerosene model emulsions. Among them, 5 strains showed high demulsifying ability, achieving 70% plus demulsifying ratio within 24 hours. Petroleum-contaminated soil, excess sludge from biological process treating oilfield produced water and sludge from municipal wastewater treatment plant were the best among all tested sources. Due to the determination limit, the colorimetric assay and blood-plate hemolysis method are not competent to screen bio-demulsifiers strains. The measurement of surface tension and oil spreading method were easy but accu- rate methods to isolate bio-demulsifiers strains. Although demulsification test of model emulsion is an effec- tive technique to target strains with the capability of breaking emulsions, it is sophisticated and time-con- suming. Thus it is recommended to use surface tension and oil spreading methods in pre-screening and vali- date the results in demulsification test with kerosene model emulsions.

Objective:To determine the law of dose variation in nasopharyngeal carcinoma patients treated with helical tomotherapy by observing the difference between the actual and planned doses of targets and at-risk organs in these patients. Methods:Ten naso-pharyngeal carcinoma patients were treated with helical tomotherapy. Each single dose distribution and the corresponding computed tomography (CT) images were transmitted to the commercial software MIMvista 6.50, which was used to perform deformable image registration on the CT images. The sum of the actual dose was then obtained by accumulating the single doses. The image-guided actu-al dose was denoted as Plan 2, and the initial plan was called Plan 1. The dose-volume histogram of the dose distribution of targets and at-risk organs in Plans 1 and 2 were compared. Results:The D98 and D95 doses of the planning gross target volume (PGTV) in Plan 2 were significantly lower than those in Plan 1. Compared with Plan 1, the Dmean and D50 doses of double parotids in Plan 2 were higher by 42.23%and 63.82%(P<0.001, P=0.001) on average, respectively, for the left parotid, as well as by 38.64%and 66.76%(P=0.002, 0.004), respectively, for the right parotid. The D2 dose of spinal cord in Plan 2 was significantly higher than that in Plan 1. The doses were higher by 16.49%on average (P=0.026). Conclusion:To achieve precise and individually adaptive radiotherapy, correcting the error between anatomy and placement during tomotherapy for nasopharyngeal carcinoma patients is necessary.