Objective To study the difference in detection rate of Mycobacterium tuberculosis in diffenent types of sumples.MethodsThe sputum,bronchial fluid,blood of 52 patients with clinical diagnosis of patients with pulmonary tuberculosis were tested with fluorescence quantitative PCR.Results45 cases were identiified in sputum and 51 cases were identified in bronehial perfusate and 15case were identified in blood.?2 was 4.875(P

Objective To develop an rapid, visuable method to detect the genotypes of HBV.Methods According to the published full-length sequence with known genotypes in GenBank, the specific primer and biotin-lablled probe were designed. We established a nucleic acid strip method for genotyping hepatitis B virus(HBV) isolates among 150 HBV infected patients and 20 healthy controls, and compared the results with those obtained by real-time PCR. Results There was 34.00% genotype B , 61.33% genotype C, 4.00% genotype B/C in 150 samples, respectively, while the remaining 0.67% unknown. The results for this assay were high comparable to the method of real-time PCR. Conclusion With the similar sensitivity and specifity when compared with real-time PCR, this rapid method is suitable to the clinical setting.

Objective To establish a rapid, sensitive, and specific quantitative method to detect hepatitis C virus. Methods A primer set targeting HCV 5'UTR was designed. The isothermal amplification was performed by the Bst DNA polymerase and AMV reverse transcriptase, under the temperature of 60℃ for 60 min. The signal was monitored by SYBR Green Ⅰ. Results One hundred and twenty positive serum samples, confirmed by the real-time PCR. All were detected by the isothermal amplification, while 110 healthy subjects' samples were negative by the both methods. The lower detect limit was determined to 10 IU/ml HCV-RNA, by the assay on serial dilutions of the quality control standards obtained from clinical investigation center of MOH. Conclusion A real time reverse loop-mediated isothermal amplification method was developed to detect HCV, with the characteristic of rapidity, high sensitivity and specificity.

0.05). Conclusions E 3/3 and the prevalence of ? 3 allele were significantly higher in Shandong elderly population.

Objective To investigate the distribution of hepatitis B virus genotypes in Hangzhou area and preliminarily identify and evaluate the applied characteristics of oligonuleotides(oligo)microarray for genotyping hepatitis B virus(HBV).Methods HBV PCR products were hybridized with oligonucleotide probes,which were prestablized on the chip.The hybridized results were colorized.According to the hybridization signal and the corresponding probe sequence,HBV genotype was determined.Results Of the 106 HBV DNA positive patients 37(34.91%)were genotype B,and 69(65.09%)were genotype C,No genotype A,E and F were found in the studied subjects.Conclusions HBV genotype B and C exsited in Hangzhou No HBV genotype A,D,E and F were found in the studied subjects.We should take further investigation for HBV genotypes by enlarging study population.The advantages of this assay are sensitive accurate,fast and economical when using it for HBV genotype test comparing with other relative methods.

Objective To investigate the distribution and virologic characteristics of HBV genotypes,and possible association with the severity of liver disease.Methods HBV genotype was determined,using oligonuleotides(oligo) microarray method.Results 37patients were differentiated as genotype B,69 genotype C.The clinical manifestations demonstrated that the serum lever of HBVDNA and HBeAg positive rate in patients of genotype C was 7.02?1.26 and 60.87% higher than 5.62?1.02and 32.43% in those of genotype B,the serum HBeAb positive rate in patients of genotype B was 67.57% higher than 39.13% in those of genotype C.The occurrence rates of those developed to chronic hepatitis B liver cirrhosis and hepatoma in those of genotype C were 46.38%,30.44% and 11.59% highter than that of 40.54%,13.51%and 8.11% in those of genotype B.Patients with genotype B were much younger than those with genotype C.Conclusions Genotype B and C exsited in Hangzhou,genotype C is a predominated genotype.The serum leverl of HBVDNA in those of genotype C is highter than that of genotype B.The positive rate of HBeAg in those of genotype B is highter than that of genotype C.The damage to liver induced by genotype C is severe than that of genotype B.

Objective To develop a rapid, accurate, specific method to detect causative agent of hand, foot and mouth disease (HFMD). Methods Specific primers and probe were designed based on highly conserved VP1 region of enterovirus 71, coxsackie virus A16 and enterovirus. The sensitivity and specificity of the real-time RT-PCR was evaluated with 35 stool samples collected from pediatric patients with suspected HFMD and 20 clinical samples from health pediatric patients. Results Out of 35 clinical samples from suspected HFMD, 35 samples were identified as positive for enterovirus, 25 clinical samples were identified as positive for enterovirus 71, 8 clinical samples were identified as positive for coxsackie virus A16, among which 3 clinical samples were identified as positive for enterovirus 71 and coxsackie virus A16. The clinical diagnostic accordance rate is 85.71%. Out of 20 clinical samples from normal pediatric patients, 5 clinical samples were identified as positive for enterovirus, 20 clinical samples were negative for enterovirns 71 and coxsackie virus AI6. Conclusion Our results indicate real-time RT-PCR offers a rapid, sensitive, specific and cheap method to detect pathogen of HFMD from clinical specimens.

Objective To investigate the association of primary liver cancer(PLC)with the mutations of HBV precore and basic core promoter(BCP)genes.Methods The serum markers of hepatitis B and the quantities of serum HBV DNA were detected in 144 HBsAg-positive PLC patients.The precore and BCP gene mutations in patients with HBeAg-negtive and HBV DNA-positive were detected by real-time PCR.One hundred and twenty chronic hepatitis B(CHB)patients were randomly selected to serve as the conol.Results There were 46(3 1.94%)patients with HBeAg-positive and 98(68.06%)patients with HBeAg-negative.In 98 HBeAg-negative patients,56(57.14%)were HBV DNA-positive,in which 43 (76.79%)were with precore 1896 gene mutations,50(89.29%)were with BCP1762/1764 gene mutations.and 38(67.86%)were with both gene mutations.Precore 1896 and BCP1762/1764 gene mutation rates in PLC patients were much higher than those in CHB patients(χ2=9.36 and 5.77,P＜0.05).Conclusion PLC may be associated with the mutations of HBV precore anti BCP genes.

Objective To investigate the clinical efficacy of warming and unblocking acupuncture in treating scapulocostal syndrome.Method Eighty patients with scapulocostal syndrome meeting the inclusion criteria were allocated by simple randomization to warming and unblocking acupuncture and conventional acupuncturegroups, 40 cases each.The warming and unblocking acupuncture group received warming and unblocking acupuncture at points Tianzong(SI11), Jianjing(GB21)and Dazhui(GV14)on the affected side and affected chest Huatuo jiaji(Ex-B2)points and uniform reinforcing-reducing acupuncture at points Quchi(LI11), Hegu(LI4)and Waiguan(TE5).The conventional acupuncture group received uniform reinforcing-reducing acupuncture at the same points as those in the warming and unblocking acupuncture group after arrival of qi.Both groupswere treated once every other day, 10 days as a course, for a total of one course.Pain severity was assessed using the Pain Visual Analogue Scale(VAS) in the two groups of patients before and after treatment.Theclinical therapeutic effects were evaluated by follow-up at one month after the completion of treatment.Result The total efficacy rate was 95.0% (38/40) in the warming andunblocking acupuncture group and 80.0% (32/40) in the conventional acupuncture group; there was a statistically significant difference (P<0.05).The PainVisual Analogue Scale(VAS) score was significantly lower in the two groups of patients after treatment than before treatment; there was a statistically significant difference (P<0.05).The VAS score was decreased more in the warming and unblocking acupuncture group after treatment and at the follow-up one month after the completion of treatment (P<0.05).Conclusion Warming and unblocking acupuncture has clinically a better therapeutic effect onscapulocostal syndrome.

BACKGROUND:The chemical compositions and structure of calcium phosphate bone cement are similar to those of human bone, which can fil the bone colapse caused by fracture and induce osteogenesis, but its degradation rate is slow. OBJECTIVE:To evaluate the biocompatibility of the calcium phosphate cement/fibrin glue and the feasibility of repairing radius defects. METHODS: In vitro cytotoxicity experiment: Mouse fibroblasts were cultured in the calcium phosphate bone cement/fibrin glue extracts, phenol solution, and RPMI 1640 culture medium containing 10% fetal bovine serum, respectively, to detect the cytotoxicity grade. Hemocompatibility experiment: Calcium phosphate bone cement/fibrin glue extracts, normal saline and distiled water were respectively added into the rabbit anticoagulation, to detect the hemolytic rate. Forty-five New Zealand white rabbits were enroled and modeled into bilateral radius defects, folowed by randomly alotted into three groups: blank control group without any intervention, experimental and control groups were given the implantation with calcium phosphate bone cement/fibrin glue and autologous radius, respectively. X-ray, histology, bone mineral density and biomechanical test were performed at postoperative 4, 8 and 16 weeks. RESULTS AND CONCLUSION:The toxicity grade of the calcium phosphate cement/fibrin glue was 0 to 1. The hemolytic rate of the calcium phosphate cement/fibrin glue was 3.15%. At 16 weeks postoperatively, X-ray showed that in the experimental and control groups, the fracture line disappeared completely, pulp cavity was recanalized, and in plastic completely. Histology showed that the reconstructed bone trabecular was obvious, plate layer of bone was mature, and medulary cavity recanalization appeared in the control group; there were a large number of new grid-shaped woven bone tissues growing into the material in the experimental group, with overt degradation, and degradation rate was in paralel to bone ingrowth. The bone density, the maximum load, maximum stress and failure energy in the experimental and control groups were significantly higher than those in the blank control group (P < 0.05), and al above indicators showed no significant differences between the experimental and control groups. These results manifest that the calcium phosphate bone cement/fiber protein glue composite material holding a good biocompatibility can promote bone tissue regeneration for bone defect repair, achieving similar curative effect with autologous bone transplantation.