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.

ObjectiveTo evaluate the feasibility and value of mini-clinical evaluation exercise (Mini-CEX) in clinical neurology practice.MethodsNinety-four interns were randomly divided into observation group and control group,students in control group were teached and managed in accordance with existing management while those in observation group were evaluated by teachers after the 1 st,2nd and 3rd week.At the end of clinical practice,all the students( including students in control group and observation group)were cross assessed by teachers based on the methods mentioned above.Results The time to complete the assessment was about 25 - 40 min.The scores of nervous system examination at the end of the training were significant different between observation group and control group and the scores of diagnosis and treatment on the basis of examination were also significant different between observation group and control group ( P ＜ 0.05 ).ConclusionThe Mini-CEX assessment and feedback to promote teaching effect is feasible in the practice process of neurology,it can make up for the deficiency of current examination.

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.

Objective To investigate the relationship between plasma homocysteine (Hey) level and ankylosing spondylitis (AS).To analyze the association between the NS,N10 methylenetetrahydrofolate reductase (MTFHR) gene polymorphism and AS.Methods One hundred patients with AS and 60 healthy controls were included in the study.The plasma Hey level was examined by enzyme-linked immunoadsorbent assay and MTHFR gene polymorphism was analyzed by the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP).Results Compared with heahhy controls,the plasma Hey level in AS patients was significantly higher than that of the controls (P＜0.01).There was no significant difference in the frequen-cies of MTHFR genotype and alleles between AS and the controls (P＞0.05),But the ratio of T/T genotype mutation was different between AS and the controls (P＜0.05).The plasma Hey level of T/T genotype was significantly higher than that of C/T or C/C genotype in AS and the controls (P＜0.01).Logisticalregression analysis indicated that Hey was an independent risk factor for AS (P＜0.01,0R=4.582,95%CI=1.984～10.585).Conclusion The plasma homocysteine level is significantly increased in AS patients.Hyperhomo-cysteinemia is an independent risk factor for AS.MTHFR T/T genotype mutation is an important mechanism of hyperhomocysteinemia and may be related with AS.

objective:To evaluate the sensitivity and predictive value of grey scale and power Doppler ultrasound assessment of bone erosionin disease activity in patients with early rheumatoid arthritis (Ra).
Methods:Fifty-six patients with early Ra underwent blinded sequential clinical, laboratory and ultrasound assessments, and at the same time 20 of these patients underwent X-ray and enhanced MRi. For each patient, 28-joint disease activity score (DaS28), erythrocyte sedimentation rate (eSR), C reactive protein (CRP) and health assessment questionnaire (haQ) were recorded. The presence of bone erosion and synovitis was investigated in 28 joints by gray-scale and power Doppler ultrasonography. The ultrasound joint count and index for active synovitis with power Doppler signal were calculated.
Results:The number of bone erosions detected by ultrasonography was 5.7 times that of X-ray, while both MRi and ultrasonography were consistent (91.5%). The number of synovitis detected by ultrasonography was 1.6 times as much as by physical examination, and consistent MRi (95.7%). PDUS parameters demonstrated a highly significant correlation with DaS28, eSR and CRP, while a negative correlation with haQ.
Conclusion:Grey scale and power Doppler ultrasonography is a sensitive and reliable method to assess bone erosion and inflammatory activity in early Ra. PDUS findings may have a predictive value in disease activity.

Objective To study the efficacy of azithromycin in the treatment of acute bronchitis. Methods The 300 patients with acute bronchitis treated by our hospital from August 2016 to February 2017 were randomly divided into the control group and the experimental group, each with 150 patients. The control group was treated with penicillin and azithromycin was treated by the experimental group. Comparative analysis was made on the clinical indicators of the experimental group and the control group. Results After the corresponding treatment, the improvement time of cough sputum in the experimental group was (4.19 ± 0.54), and the improvement time was (4.32 ± 0.65). The improvement time of cough sputum in the control group was (6.29 ± 0.87), and the improvement of pharyngeal pain was (5.32 ± 0.76) days. However, the improvement time of cough expectorus and pharyngeal pain in the experimental group was significantly shorter than the control group, the differences were statistically significant (P＜0.05). Of the patients in the experimental group, 6 patients were not effective, 14 patients improved, 70 patients showed positive results and 60 patients recovered. In the control group, the total number of cases was 110, 15 were invalid, 25 were improved, 60 were developed and 50 were cured. However, the efficiency of the experimental group was 86.67 %, significantly higher than that in the control group (73.33 %), the differences were statistically significant (P＜0.05). The clinical recovery time of the patients in the experimental group was (5.65±1.19), which was significantly shorter than that of the control group (6.89±1.46), the differences were statistically significant (P＜0.05). Conclusion The clinical efficacy of azithromycin in the treatment of acute bronchitis is better, which can significantly improve the treatment efficiency, relieve the patient’s related symptoms, and have the significance of clinical promotion.

Objective To observe the effect of resveratrol on the apoptosis and expressions of bal-2 and bax protein in articular chondrecytes of rabbits experimental osteoarthritis (OA) model,and further explore the mechanisms of resveratrol in the treatment of OA.Methods Thirty Newzealand rabbits were randomly divided into 5 groups：group A (normal control group),group B (model control group),group C (resveratrol intervention high dosage group),group D(resveratrol intervention middle dosage group),group E (resveratrel intervention low dosage group).The model of OA was established with Hulth's modeling method in group B,C,D,E.Four weeks later,groups A and B received intragastric administration of distilled water containing 0.1% DMSO daily and group C,D,E received intragastric administration of resveratrol solution daily (concentration was 60 mg/ml) in different dosages for 6 weeks.Daily dosages of group C,D,E were 120,60,30 mg·kg-1·d-1,respectively.Then the rabbits were sacrificed and the cartilage sections of right femoral medial condyle were analyzed by immunohistochemistry for bcl-2 and bax,TUNEL for apoptosis.Results ① The apoptosis rates of chondrocytes in group B,C,D,E were significantly higher than those in group A (P＜0.01).The apoptosis rates of chandrocytes in group C,D,E were decreased compared with those in group B (P＜0.05).②The positive rates of bcl-2 and bax expression in chondrocytes in group B were significantly higher than those in group A (P＜0.01),but the ratio of the positive rate of bcl-2 expression to that of bax in group B was lower than that in group A (P＜0.01).The positive rates of bcl-2 expression in chondrocytes in group C,D,E were much higher compared with those in group B (P＜0.01).The positive rotes of bax expression in chondrocytes in group C,D,E were lower compared with those in group B (P＜0.01).The ratio of the positive rate of bcl-2 expression to that of bax was increased in group C,D,E compared with group B (P＜0.01).Conclusion Resveratrol can suppress the excessive apoptosis of chondrocytes in experimental OA by up-regulating the expression of bcl-2 while down-regulating the expression of bax and improving the ratio of bcl-2 to bax .Suppressing the excessive apoptosis of chondrocytes in experimental OA may be one of the mechanisms for resveratroi's effect in the treatment of osteoarthritis.

Digoxin plays a part in healing of congestive heart failure in clinic. Its therapeutic dose is very approximate to toxic dose and even they overlap each other sometimes. There are many influencing factors on blood drug level of digoxin. Pharmacodynamics and pharmacokinetics varies with different individuality. It is indispensable to detecting blood drug level in order to treat disease and prevent intoxication. Integrating with the detecting-methods of blood drug level of digoxin home and broad, characteristic of many methods are summarized from sensitivity, linearity range, cross-reaction and precision. These methods include radio immunoassay, enzyme immunoassay, chemiluminescence immunoassay, fluorescence immunoassay and HPLC-MS-MS. These methods are popular for their specialized ascendancy. The cost of radio immunoassay is low. Enzyme immunoassay has good specificity. Sensitivity and stability of chemiluminescence immunoassay is very excellent. Fluorescence polarization immunoassay is sensitive and convenient. HPLC-MS-MS has high resolution and good specificity. One of the development tendencies is to combine two or more methods in detecting the blood drug level of digoxin which contribute to these methods integrated use.
Chemistry Techniques, Analytical ; methods ; Chromatography, High Pressure Liquid ; Digoxin ; blood ; Enzyme-Linked Immunosorbent Assay ; Fluorescence Polarization Immunoassay ; Fluoroimmunoassay ; Humans ; Radioimmunoassay ; Reproducibility of Results ; Tandem Mass Spectrometry

When the glomerular mesangial cells of rats were cultured in vitro at high glucose concentration,the activity of extraeellular signal-regulated kinase(ERK),the expression of transforming growth factor-?_1(TGF-?_1)mRNA and the content of typeⅣcollagen in the supematant were higher than those at normal glucose concentration.These effects were inhibited by fluvastatin.The results showed that the activation of ERK signal transduction pathways appeared to play a role in the onset and progression of diabetic nephropathy. Furthermore,fluvastatin could protect the kidney by inhibiting ERK signal transduction pathway and TGF-?_1 expression.

OBJECTIVETo study the genotype distribution of extended-spectrum beta-lactamases (ESBLs) in ESBLs-producing Escherichia coli (E. coli) isolates from posthepatitic cirrhosis' patients with bloodstream infection.

METHODSE. coli were isolated in bloodstream from patients with posthepatitic cirrhosis between January and December in 2011. The strains were identified by VITEK-II. The antibiol susceptibility tests were performed with K-B method. beta-lactamases genes were detected multi-PCR, PCR, sequence and blast.

RESULTSA total of 79 non-duplicate clinical isolates of E coli were consecutively collected from liver cirrhosis' patients with bloodstream infection. There were 20 isolates produced TEM-1 type beta-lactamases and 1 isolate produced SHV-1 typebeta-lactamases. 40 clinical isolates were detected to produce CTX-M type ESBLs, there were 20 CTX-M-1 group and 26 CTX-M-9 group, including 6 stains habouring both CTX-M-1 and CTX-M-9 group. Eight CTX-M genotypes were confirmed by sequencing of the PCR products, including CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, CTX-M-28, CTX-M-31, CTX-M-65 and CTX-M-79.

CONCLUSIONCTX-M genotype ESBLs was the most popular extended-spectrum beta-lactamases in E. coli isolated from liver cirrhosis' patients with bloodstream infection. The CTX-M-14 is the dominant epidemic type.

Bacteremia ; microbiology ; Cross Infection ; microbiology ; Drug Resistance, Bacterial ; Escherichia coli ; drug effects ; enzymology ; genetics ; isolation & purification ; Escherichia coli Infections ; microbiology ; Escherichia coli Proteins ; genetics ; Genotype ; Hospitalization ; statistics & numerical data ; Humans ; Liver Cirrhosis ; therapy ; Microbial Sensitivity Tests ; beta-Lactamases ; genetics ; metabolism