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

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

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

OBJECTIVETo investigate the clinical effect of the nervus cutaneus femoris posterior pedicle flap on repairing large soft tissue defects at the heel or inferior segment of the shank.

METHODSTotally 14 cases were followed up for 8-22 months (mean 15.5 months) to observe the clinical effects of nervus cutaneus femoris posterior pedicle flap on repairing large soft tissue defects of the heel or inferior segment of the shank. Among them, there were 3 patients afflicted with infection and cutaneous defects in the middle and inferior segment of the shank after internal fixation of open fracture, 4 patients with soft tissue defects of the ankle and uncovered tendo calcaneus, and 7 patients with soft tissue defects of the heel and exposed calcaneus.

RESULTSThe flaps survived well in 13 cases and partial necrosis occurred in 1 case that was thereafter cured with changing dressing. Various extents of pain and stiffness of the knee joints were present in all cases and disappeared through 1-8 weeks' (mean 3.2 weeks) functional exercises. The last follow-up showed that all the flaps kept good texture and satisfactory appearance.

CONCLUSIONSThe nervus cutaneus femoris posterior pedicle flap, having the advantages of simple surgical procedures, anastomosing the nerves and restoring the sensation of recipient site, can be used for recovering large soft tissue defects of the shank and ankle.

Adolescent ; Adult ; Aged ; Female ; Heel ; surgery ; Humans ; Leg ; surgery ; Male ; Middle Aged ; Soft Tissue Injuries ; surgery ; Surgical Flaps

OBJECTIVETo analyze the clinical feature of temporomandibular joint (TMJ) ankylosis and to investigate the advantage and disadvantage of all kind of surgery methods of TMJ ankylosis for supplying references to the treatment of TMJ ankylosis.

METHODSAetiology, sex, age at time of treatment, clinical features, radiographic findings, surgical treatment was reviewed and analyzed.

RESULTSTrauma (66.86%, n = 117) and infection (24.57%, n = 43) were the primary causes of TMJ ankylosis. The 10 approximately 19 years age group (44.57%, n =78) occupied the highest frequency in five age groups, one hundred and fourteen (65.14%, n = 114) of the patients presented with lateral ankylosis. They were carried out different surgery method according to their difference of age, aetiology, and so on.

CONCLUSIONSThe person at 10 approximately 19 years old had the more feasibility of TMJ ankylosis than others. Trauma was the commonest cause of ankylosis. Different surgical method should be choose according to the different patient.

Adolescent ; Adult ; Ankylosis ; surgery ; Child ; Child, Preschool ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Osteotomy ; Retrospective Studies ; Temporomandibular Joint Disorders ; surgery ; Young Adult

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.