Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective To evaluate the expression of Toll-like receptor(TLR)on the monocyte- derived dendritic cells(DC)from chronic hepatitis B(CHB)patients and to analyze the expression pro- file and significance of the TLR such as TLR3,TLR4,TLR?,TLR8 and TLRg,which are associat- ed with immune response to viral infection.Methods Peripheral blood mononuclear cell(PBMC) centrifugated by the hydroxyethyl starch(HES)centrifugation were cultured and induced into DC by granulocyte-maerophage colony stimulating factor(GM-CSF)and interleukin-4(IL-4),and their mor- phology and phenotype were detected by the inverted microscope and flow cytometry respectively. Monocyte-derived DC were obtained from 10 chronically hepatitis B virus(HBV)-infected patients and 15 healthy volunteers.TLR3,TLR4,TLR7,TLRS,TLR9 expression on immature and mature DC were analyzed by FACS Calibur.DC was pulsed with HBcAg on day 3 and 5,then DC maturation and ability to process HBcAg and to stimulate autogeneic T cells were evaluated.Results Monocyte- derived DC developed different TLR expression patterns as they went through different maturation stages.TLR7,TLR8 expressions on immature DC and TLR3,TLR7 expressions on mature DC were lower in CHB than in control(for TLR7,TLR8 expression on immature DC:75.9%,1.0%vs 98.4%,15.4%,P

Background and Objective: With the application of laparoscopy,laparoscopic gastrectomy for the treatment of patients with early gastric cancer has been performed,but the safety and effectiveness of this method needs to be explored.This study evaluated the safety and effectiveness of laparoscopy-assisted and conventional open distal gastrectomy for patients with early gastric cancer.Methods: A search of MEDLINE,EMBASE,the Chinese Biomedical Database(CBM),and Cochrane Central Register of Controlled Trials(CENTRAL)identified all the randomized clinical trials that compared laparoscopy-assisted gastrectomy with open distal gastrectomy for patients with early gastric cancer published in the last 10 years.Quality assessment was done on each trial and relevant data were extracted from qualified trials.Meta-analysis was performed using RevMan 4.2.2 software(Cochrane).Results: Six randomized controlled trials(RCTs)involving 218patients were included.Comparing laparoscopic resection with open resection,results showed less estimated blood loss[WMD(weighted mean difference):-121.86; 95% CI(confidence interval):-145.61,-98.11 ;P<0.001],earlier postoperative first flatus(WMD:-0.95; 95 % CI:-1.09,-0.81 ; P<0.001),and shorter durations of hospital stays(WMD:-2.27;95% CI:-3.47,-1.06; P=0.0002),but longer surgery times(WMD:58.71 ; 95 % CI: 52.69,64.74; P<0.001)and fewer lymph nodes dissected(WMD:-3.64; 95% CI:-5.80,-1.47; P=0.001).There was no significant difference between the two groups in postoperative complications[OR(odds ratio): 0.57; 95% CI: 0.31,1.03; P=0.06].Conclusion: The short-term outcome of laparoscopy-assisted distal gastrectomy for patients with early gastric cancer is superior to the open procedure,but its long-term outcome should be proven by further outcomes of RCTs.

OBJECTIVETo study the strategy of the treatment for dislocation of cervical vertebra.

METHODSThe clinical data of 39 cases with dislocation of cervical vertebra were analyzed. Among them,29 were male and 10 were female. The average age was 40 years old (range from 6 to 74 years old). Segment of dislocation: 15 cases in C(1,2), 1 case in C(3,4), 9 cases in C(4,5), 9 cases in C(5,6), 5 cases in C(6,7). Spinal injury according to Frankel grade, 9 cases were A grade,8 were B, 5 were C, 8 were D, 8 were E, 1 case had radicular symptom. Thirty-two cases were early and rapidly treated with traction (progressive weight). Seventeen cases were treated with operation.

RESULTSTraction-reduction was successful in 90% of patients. According to Frankel grade, 32 cases averagely improved 0.63 grades. Six cases of severe spinal injury accompany with interlocking of zygopophysis died.

CONCLUSIONInspecting weight of traction is important in rapid traction-reduction for dislocation of cervical vertebra. The choice of surgical treatment depends on the degree of reduction, the result of MRI,the grade of spinal trauma and the status of patients.

Adolescent ; Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Child ; Female ; Humans ; Joint Dislocations ; surgery ; therapy ; Male ; Middle Aged ; Traction ; Treatment Outcome ; Young Adult

OBJECTIVESTo observe immunogenicity of new DNA vaccine encoding for hepatitis B virus core antigen (HBcAg).

METHODSA new DNA vaccine (pSW3891/HBc) encoding for hepatitis B virus core antigen was constructed using plasmid pSW3891 which can be used in human. Control and experiment groups of Balb/c mice were immunized with pSW3891 or pSW3891/HBc by gene gun. Anti-HBc in sera of mice was tested by ELISA (enzyme linked immune sorbent assay). Specific cytotoxicity of cytotoxic T lymphocyte (CTL) of mice was detected by LDH release assay.

RESULTSpSW3891/HBc can express in 293T cell line effectively. Mice immunized with pSW3891/HBc showed strong anti-HBc response and specific high cytotoxicity of CTL.

CONCLUSIONpSW3891/HBc induced significantly humoral and cellular immune responses in Balb/c mice.

Animals ; Hepatitis B Antibodies ; blood ; Hepatitis B Core Antigens ; genetics ; immunology ; Hepatitis B Vaccines ; immunology ; Immunity, Cellular ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccines, DNA ; immunology

Objective To study on epidemiologic characteristics of human plague cases in Sanjiangyuan Area,and provide theoretical basis to work out the preventive measures.Methods Based upon the epidemiology information from the human plague case data bank of Qinghai Province,human plague data were analyzed retrospectively in Sanjiangyun Area by sorting,verifying and summing up of these data,including some of case file and monitoring data.Results Except for 12 years in the period of 1960 to 2006,there were human plague cases happened every year.The morbidity occurred mainly in 12 counties of 4 states,including Yushu,Guolou,Huangnan and Hainan,and Tanggula Town of Geermu City,a total of 85 human plague episodes were occurred,resulting 238 onsets,134 deaths,and a matality rate of 56.30%.The sources of infection were respectively Himalayan mormot 27.31%(65/238),artiodactyls 14.71%(32/238),carnivora 2.10%(5/238),Lagomorpha 0.42%(1/238),the pneumonic plague patient 49.16%(117/238),and biting of flea 6.30%(15/238).The prevalent season was from May all the way to November,the peak-months were August and September.After October,the sheep as the source of infection initiating human being plague accounted for 23.53%.Among the clinical types,the most prevalent type was pneumonic type(61.34%),and the rest,glandular type(17.23%),septic type(16.81%)and other types(4.62%),but the first plague case in each epidemic was mainly the glandular plague.Conclusions In recent years,the tendency of human plague prevalence increases in Sanjiangyuan Area,it is urgent to improve and adjust the prevention and treatment measures in time.

This study was aimed to investigate the distribution feature of HLA-DR/DQ gene linkage disequilibrium in Chinese Han population and to improve the accuracy of HLA matching results. Genotyping of HLA-DR and HLA-DQ gene locus was performed using PCR-SSP typing in Chinese Han population receiving kidney transplantation. The results showed that there were 29 new linkage combinations in 1799 patients, in which DR13-DQ5, DR11-DQ8 and DR8-DQ8 were discovered for 11, 8 and 7 times respectively while DR9-DQ8, DR12-DQ6 and DR14-DQ4 were both discovered for 6 times. The linkage disequilibrium parameters of these haplotypes were negative, showing that these linkages were uncommon. It is concluded that this study not only enriches the classical HLA-DR/DQ linkage combinations, but also indicates the national relevance of combination distribution, and it has great importance in improving the accuracy of HLA matching experiments and reducing unnecessary repeated work.
Adult ; Asian Continental Ancestry Group ; genetics ; Gene Frequency ; Genotype ; HLA-DQ Antigens ; genetics ; HLA-DR Antigens ; genetics ; Haplotypes ; Humans ; Kidney Transplantation ; Linkage Disequilibrium

OBJECTIVETo determine and compare the content of luteolin-7-O-beta-D-glucoside and apigenin-7-O-beta-D-glucoside in Flos Chrysanthemi from different collection time, sources, grades and processes.

METHODThe contents were determined by RP-HPLC. Zorbax SB C18 column (4.6 mm x 250 mm, 5 microm) was used as analysis column, the mobile phase was acetonitrile-pH 2.0 phosphate buffer solution with gradient elution, the detector was set at 338 nm.

RESULTThe contents of two components changed at some degree in Flos Chrysanthemi from different collection time, different plant sites or with different grades, while the contents varied obviously among Flos Chrysanthemi from different source and different sorts. No obvious difference was found in Flos Chrysanthemi from different year.

CONCLUSIONThe contents of two components were influenced by process, plane site, source and sorts, especially by source and sorts.

Apigenin ; analysis ; China ; Chrysanthemum ; chemistry ; Drug Compounding ; Ecosystem ; Flowers ; chemistry ; Glucosides ; analysis ; Luteolin ; analysis ; Plants, Medicinal ; chemistry ; Seasons

Objective To examine the association between self-assessed somatotypes and depression related symptoms.Methods The study sample included 5555 students aged 9 to 18 years who had attended the Physical Fitness and Health Surveillance of Chinese School Students(2010)in Chizhou city,Anhui province.Association between self-assessed somatotypes,body mass index(BMI)and depression symptom were examined.Results There was a slight consistency between self-assessed somatotypes and BMI in both boys and girls(Kappa=0.217,P=0.000; Kappa=0.203,P=0.000).Significant difference in the prevalence of depression was found among weight misperception groups in both genders(x2=145.223,P=0.000).The prevalence of umderestimation of somatotypes was significantly higher in boys than in girls,while the result was reversive on the prevalence of overestimation.Additionally,the prevalence of depression was higher in girls than in boys(x2=5.199,P=0.023).Through logistic regression,data showed that self-assessed somatotypes and miscalculated groups were significantly associated with depressive symptoms.Compared to the group that self-assessment as being normal,those students when self-assessed as being slim,overweight or obesity were more likely to be depressive,with odds ratio(ORs)as 1.255(95％CI:1.066-1.478),1.538(95％CI:1.275-1.856)and 1.713(95％ CI:1.035-2.834),respectively.Overestimated and underestimated somatotypes appeared to be risk factors causing symptoms of depression(OR=1.705,95％CI:1.382-2.105; OR=1.241,95％CI:1.059-1.454).Conclusion Slight consistency was found between self-assessed somatotypes and BMI,while the misjudged somatotypes were the risk factor related to depressive symptoms.It was suggested that life skills education should be carried out as preventive intervention approach,to improve the physical and mental health well-being of children and adolescents.

Objective To investigate the association between psychological sub-healthy status and risks on injuries and related predictors among adolescents in China,in order to develop reasonable intervention programs.Methods Adolescents were selected from middle schools and colleges in eight cities.Baseline status on sub-healthy that related to psycho-pathology was measured using the Multidimensional Sub-health Questionnaire of Adolescents (MSQA).Data on Self- or unintentional injuries were collected using a standardized injury surveillance questionnaire every 3 months,for three times (T0,T1,T2 and T3).Results A total of 12 113 students completed the study.Prevalence rates related to self-injury among boys and girls were 19.4％ and 19.9％,respectively.The rates of having more than 4 types of self-injuries in boys and girls were 4.7％ and 4.2％,respectively.Boys had a significantly higher unintentional injury rate than that of girls (62.2％ vs.57.3％ ).The differences in the rates of having more than 4 types of unintentional injury between boys (3.8％) and girls (3.4％) were also statistically significant.Rates on self- and unintentional-injuries increased in T1,T2 and T3 when the psychological symptoms had an increase.Data from Multiple regression analysis demonstrated that the baseline psychological sub-healthy status was a predictive factor for higher risk of self- and unintentional- injuries at each of the follow-up waves.Conclusion Sub-healthy status of psycho-pathology was associated with an increased risk on self-and unintentional injuries among adolescents.Integrative psycho-pathological and behavioralintervention programs might help in reducing the risk of injuries among the Chinese adolescents.