Objective To amplify the 16S RNA fragments of 7 clinically isolated strains of Brucella spp. by PCR-RFLP technique, so as to provide experimental basis for the studies on diagnostics, genetics and epidemiology of Brucella spp. Methods According to the gene sequence of ATCC 25840 standard strain in GenBank, special primers for the 16S RNA conservative area in the Brucella spp. were designed. DNA extraction and PCR amplification of the 16S RNA fragments were performed with the 7 isolated strains. PCR products were then sequenced and RFLP analysis was conducted with appropriate restricted enzymes to study the homology and the mutation sites in those strains. Meanwhile, the clinical data of infected patients were retrospectively analyzed to evaluate the relationship between the clinical features and genotypes of Brucella infection. Results The amplified target fragments were about 1500bp in length and consistent with what was expected. The sequencing and homology analysis showed a 98.88% homology and 11 mutation sites among the 7 isolated strains. Four genotypes were identified by RFLP. Retrospective analysis of the clinical data indicated that no obvious relationship existed between the genotypes and the clinical features. Conclusions Amplifying 16S RNA fragments by PCR technique is a feasible method to make an early diagnosis of Brucella infection. The 7 clinically isolated strains are different in genotypes and 16S RNA fragment is a highly conservative fragment in bacterial genome with some mutations. The research provides evidence for the genetics and epidemiology of brucellosis.

Calcium Channel Blockers ; pharmacology ; Cell Survival ; drug effects ; Drug Antagonism ; Humans ; Kidney ; cytology ; drug effects ; Lead ; toxicity

Adult ; Genotype ; Humans ; Male ; Middle Aged ; Peptidyl-Dipeptidase A ; blood ; genetics ; Pneumoconiosis ; blood ; genetics ; Polymorphism, Single Nucleotide

Adult ; Aged ; Carcinoma, Squamous Cell ; surgery ; Esophagus ; surgery ; Humans ; Laryngeal Neoplasms ; surgery ; Laryngectomy ; Male ; Middle Aged ; Phonation ; Postoperative Period ; Reconstructive Surgical Procedures ; methods ; Speech, Alaryngeal ; Trachea ; surgery

Objective To analyze the mutations of GATA4 gene in Han Chinese patients with atrioventricular septal defect(AVSD)and investigate the association between GATA4 gene and pathogenesis. Methods Ninety-four Han Chinese patients with AVSD were recruited,including 23 patients with Down syndrome and 71 patients without. One hundred healthy age-matched Han children were used as the control. Blood samples were drawn. Encoding region and flanking introns of GATA4 gene were amplified using polymerase chain reaction. The mutations were detected by DNA fragment sequences analysis. Results Three novel missense mutations(c.106C ＞ G,p.P36A;c.259C ＞ T, p.P87S;c.504C ＞ A,p.D168E)of the GATA4 gene were identified in three patients with complete AVSD without Down syndrome,and a fourth novel missense mutation(c.1079A ＞ G,p.E360G)was noted in a patient with complete AVSD and Down syndrome. A polymorphism of the GATA4 gene(c.G99T,p.A33A)from six patients was detected. Conclusions The GATA4 gene might be involved in the etiology of AVSD by functional changes resulting from gene mutation. The low incidence of GATA4 gene mutations in patients with AVSD with or without Down syndrome might suggest that AVSD is a polygenetic disorder.

Objective To monitor the constituents and resistant tendency of bacterial pathogens isolated from diarrheal patients in our hospital form 1994 to 2005 to offer the basis for guiding epidemiologic study,vaccination research and clinical treatment. Methods Enteric pathogenic bacteria were cultured and identified to species,group and serotype with biochemical and serologic methods and the susceptibility of bacteria to antimicrobial agents were tested. Results Enteric pathogenic bacteria were isolated predominantly in male patients and mainly in children and youngsters. It reached a peak from July to September every year. Shigella spp.(75.11%) was the most frequendy isolated pathogens and followed by Vibrio spp.(12.7%),Salmonella spp.(6.28%),Aeromonas spp.(4.43%) and Escherichia coli(1.25%).During the period from 1994 to 2005,diarrheal pathogens had a trend of decrease especially Shigella spp.and Salmonella spp.. Of the 6329 isolates of Shigella spp., 75.62% was S. flexneri and S.soanei,S.dysenteriae and S. boydii constituted 23.98%,0.22% and 0.01% respectively.The sensitivity of different species,group or serotype to different antimicrobial agents was not the same.S.flexneri and Aeromonas spp. were highly resistant to most of antibiotics. However, S.sonnei and Vibrio spp.had good susceptibility to antibiotics tested except trimethoprim/sulfamethoxazole and ampicillin. Conclusion There are many species and serotypes of enteric pathogenic bacteria causing infective diarrhea and the distribution changes gradually in Beijing. The resistance rate of enteric pathogenic bacteria to antibiotics is not the same in different species and serotypes.so strict surveillance iS always needed.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Epstein-Barr Virus Infections ; Female ; Herpesvirus 4, Human ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; pathology ; virology ; Lymphoma, T-Cell, Peripheral ; drug therapy ; pathology ; Neoplasms, Second Primary ; drug therapy ; pathology ; Prednisone ; therapeutic use ; Vincristine ; therapeutic use

Genetic transformation is an effective method to improve breeding objective traits of orchids. However, there is little information about genetic transformation of Cymbidium sinensis. Rhizomes from shoot-tip culture of C. sinensis cv. 'Qijianbaimo' were used to establish a practical transformation protocol of C. sinensis. Pre-culture time, concentration and treating methods of acetosyringone, concentration of infection bacteria fluid (OD600), infection time, and co-culture time had significant effects on β-glucuronidase (GUS) transient expression rate of C. sinensis cv. 'Qijianbaimo' rhizome. The GUS transient expression rate of rhizome was the highest (11.67%) when rhizomes pre-cultured for 39 d were soaked in bacterium suspension (OD600 = 0.9) supplemented with 200 μmol/L acetosyringone for 35 min, followed by culturing on co-culture medium supplemented with 200 μmol/L acetosyringone for 7 d. Under this transformation conditions, 3 transgenic plantlets, confirmed by GUS histochemical assay and PCR, were obtained from 400 regenerated plantlets, and the genetic transformation rate was 0.75%. This proved that it was feasible to create new cultivars by the use of Agrobacterium-mediated genetic transformation in C. sinense.
Agrobacterium ; Coculture Techniques ; Genetic Engineering ; Glucuronidase ; Orchidaceae ; genetics ; Plants, Genetically Modified ; genetics ; Polymerase Chain Reaction ; Transformation, Genetic

Adult ; Antigens, CD20 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; CD79 Antigens ; metabolism ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; Male ; PAX5 Transcription Factor ; metabolism ; Young Adult

AIM: To investigate the early changes of retinal function in diabetic patients detected by multifocal electroretinogram (mfERG).METHODS: The first-order kernel responses of mfERG were recorded from eyes of 33 normal control subjects,63 diabetic patients without retinopathy and 43 diabetic patients with background retinopathy. The response densities and implicit times of N1 and P1 were compared among the control, diabetic patients without retinopathy and diabetic patients with retinopathy.RESULTS: The response densities of N1 and P1 in central 3 rings were reduced significantly in diabetic eyes with and without retinopathy. And the implicit times of N1 and P1 were delayed significantly only in diabetic eyes with retinopathy.CONCLUSION: mfERG can detect the early changes of retinal function quantitatively in diabetic patients. Analysis of response densities and implicit times of N1 and P1 can reflect the progress of local retinal dysfunction in diabetes.