OBJECTIVETo investigate the relationship between penA/ponA and penicillin resistance of Neisseria gonorrhoeae.

METHODSAgar dilution method was used to determine the minimum inhibitory concentrations (MICs) of the strains. Polymerase chain reaction-single stand conformation polymerphism (PCR-SSCP) and PCR- restriction fragment length polymorphism (RFLP) were used to detect the mutations in ponA and penA genes, which encoding the penicillin binding protein-1 and -2 (PBP1 and PBP2), respectively.

RESULTSAll the 80 N. gonorrhoeae isolates had a D345 insertion detected in penA while 93.7% of N. gonorrhoeae isolates having a point mutation Leu421 --> Pro in ponA. Most of the penicillinase producing N. gonorrhoeae (PPNG) strains possessed the mutations in ponA and penA.

CONCLUSIONOur data suggested that the plasmid and chromosome mediated penicillin-resistance conjugately increased the level of resistance.

Amino Acid Sequence ; Bacterial Proteins ; genetics ; Base Sequence ; DNA Mutational Analysis ; DNA, Bacterial ; Genes, Bacterial ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Mutagenesis, Insertional ; Mutation ; Neisseria gonorrhoeae ; drug effects ; genetics ; Penicillin Resistance ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational

OBJECTIVEUsing molecular epidemiology methods to investigate relationship between genotypes and drug-resistance of neisseria (N.) gonorrhoeae in Shanghai area.

METHODSA random amplified polymorphic DNA (RAPD) fingerprint method at the molecular level was used to differentiate the strains which were isolated from the outpatients of sexually transmitted disease clinics. The sensitivity to antibiotic of the 78 N. gonorrhoeae strains on 9 different antibiotics was tested and the relationship between different genotypes and phenotypes was studied.

RESULTSSelected RAPD primer could give out a group of amplification polymerase chain reaction bands with some main segments common to all the N. gonorrhoeae strains tested and some segments were different among the N. gonorrhoeae strains. All the 78 N. gonorrhoeae strains could be classified as three different groups (I, II and III). The strains could also be distinguished as four types (A, B, C and D) according to drug-resistance status. Using correspondence analysis method, the relationship between the three genotypes and four resistance types could be identified.

CONCLUSIONRAPD fingerprint seemed a useful genotyping method and could be used for molecular epidemiological studies.

Adolescent ; Adult ; Anti-Bacterial Agents ; pharmacology ; China ; epidemiology ; DNA Fingerprinting ; DNA, Bacterial ; genetics ; Drug Resistance, Bacterial ; Female ; Genotype ; Gonorrhea ; epidemiology ; microbiology ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Molecular Epidemiology ; Neisseria gonorrhoeae ; classification ; drug effects ; genetics ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo explore the role of toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in myocardial ischemia/reperfusion injury (MI/RI) by observing the dynamic expression changes at mRNA and protein levels early after myocardial ischemia/reperfusion (I/ R).

METHODSThe Wistar rats were randomly divided into Sham and I/R group (n = 42), and killed according to different reperfusion time (1, 2, 4, 6, 12, 24 h and 7 d). Structural and morphous changes of myocytes were observed under optical microscope. The mRNA and protein levels of TLR2 and TLR4 were detected using real-time PCR (RT-PCR). Monocyte chemokine protein-1 (MCP-1) and interleukine-6 (IL-6) mRNA levels were measured by reverse transcriptase-polymerase chain reaction (rt-PCR).

RESULTS(1) With the extension of reperfusion time, the myocardial infarct size increased smoothly, and reached the plateau at 4 h, then stayed in the platform. After reperfusion for 7 d, the ventricular had been remodeled. (2) At the beginning of reperfusion, myocardial structure showed no significant change in Sham group, but had different degrees of injury in I/R group. In rats of the group reperfused for 7 d the left ventricular remodeling could be visible. (3) Compared to sham group,TIR2, TLR4, MCP-1, IL-6 mRNA level were increased in myocardium in I/R group. TLR2 and TLR4 both peaked at 4 h of reperfusion, IL6 peaked at 6 h, followed by a gradually decrease. TLR4 and IL-6 mRNA levels rose again at 7 d. MCP-1 level in I/R group remained fairly with sham group at the beginning of reperfusion, and markedly elevated at 7 d.

CONCLUSIONExpression of TLRs mRNA in myocardium during early after myocardial ischemia/reperfusion increased rapidly and activated TLRs might play an important role in MI/RI through promoting the generation of inflammatory factors. At the late reperfusion, TLRs levels raise again and the expression of inflammatory factors increase once again, Those may probably affect the remodeling of ventricular, and injure myocardial structure and function.

Animals ; Chemokine CCL2 ; metabolism ; Disease Models, Animal ; Interleukin-6 ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; Rats ; Rats, Wistar ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism

OBJECTIVETo set up random amplified polymorphic DNAs (RAPD) method in genotyping Neisseria gonorrhoeae on DNA level, and to explore its use to trace the source of infection.

METHODSFour different pretreatments were used to extract the Neisseria gonorrhoeae genomic DNA with its advantages and disadvantages compared. Arbitrary sequence was then used to amplify the genomic DNA of Neisseria gonorrhoeae and RAPD fingerprint maps was applied to distinct the Neisseria gonorrhoeae strains. Finally, RAPD fingerprint of Neisseria gonorrhoeae strain between patient and his/her sexual partner was compared.

RESULTSCetyltrimethylammonium bromide method was classical in extracting genomic DNA, and could get integrated genomic DNA and good fingerprint maps, since main segments were common to all the Neisseria gonorrhoeae but some were different among strains so that the fingerprint of different Neisseria gonorrhoeae were distinctive. However, fingerprint maps of Neisseria gonorrhoeae collected from sex partners were quite similar.

CONCLUSIONBased on genomic levels, effective fingerprint maps could be identified and to classify the Neisseria gonorrhoeae into different genotypes. RAPD fingerprint maps could be used to trace the source of infection.

DNA Fingerprinting ; DNA, Bacterial ; analysis ; Genotype ; Humans ; Neisseria gonorrhoeae ; classification ; genetics ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo explore the relationship between myeloperoxidase (MPO) 129 A/G promoter polymorphisms and the severity of coronary artery disease (CAD).

METHODSThe study enrolled 267 patients who had been diagnosed as coronary artery diseases by coronary angiography. The serum MPO activity was detected by colorimetric method. PCR-RFLP method was used to decide the genotypes of the patients. The severity of CAD was evaluated by the numbers of stenotic coronary arteries and the Gensini scores respectively.

RESULTSThe MPO 129 locus G and A alleles frequency were 0.893 and 0.107, respectively. No significant difference was observed in serum MPO activity between different genotypes (P > 0.05). The distribution of genotypes in different Gensini score groups had no significant difference (P > 0.05). The patients with GG genotypes were prone to develop mutivessel diseases.

CONCLUSIONNo significant correlation exists in MPO 129 locus polymorphism and serum MPO activity. The MPO 129 locus polymorphism isn't a reasonable predict factor of CAD severity.

Adult ; Aged ; Aged, 80 and over ; Coronary Angiography ; Coronary Artery Disease ; blood ; genetics ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Peroxidase ; blood ; genetics ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Risk Factors ; Severity of Illness Index

Objective Both epidemiologic characteristics and transmission mode of a varicella outbreak among schoolchildren in Shenzhen city were studied and related control measures were discussed.Methods Case definition was established.Case-finding and face-to-face investigation were conducted,followed by analysis on distributions of time,place and persons of the outbreak.Association between possible modes of transmission and the outbreak was explored.Retrospective cohort study was carried out.Results The overall attack rate of this outbreak was 4.4％ (122 of 2742).The attack rate among primary schoolchildren (8.3％,118/1419) was higher than that in the middle-school children (0.3％,4/1323).There were no statistically significant differences on the attack rates between male and female students.A total of 22 classes from the 9 grades were affected by this outbreak and the aggregation of varicella cases was found in classes.The highest attack rates was found in the students of fifth grade (23.7％),followed by from the third grade (13.4％).The main transmission mode appeared to be close personal contact.The outbreak,with four peaks of incidence,lasted 72 days.Data from the investigation suggested that the primary case was introduced into the school during military training involved by the students.Activities related to physical examination for all the schoolchildren seemed to have contributed to the spread of this disease among classes.Delayed isolation of cases appeared to be the major factor causing the spread of disease among classes.Intensive off-school activities or free community bus for children did not seem to be associated with the spreading of the disease.52.5％ (64/122) of the cases had received one-dose of varicella vaccine,with the median of onset after the vaccination as 7 years.The results demonstrated that one single-dose vaccine or long vaccination period were factors related to the insufficient immunity that causing the outbreak of disease.Conclusion Varicella outbreak has been one of the most grievous public health problems in schools,posting challenges on the implementation of isolation measures,in particular.Once the chain of transmission is in place,it is difficult to make the universal measures effective within a short period.

OBJECTIVETo evaluate the risk of human infection after the outbreak of avian influenza H5N1 in animals, and probe the possibility for virus transmission.

METHODSBy means of field epidemiological study, molecular epidemiology, serology and emergency surveillance, persons who had ever closely contacted with sick or dead poultry were observed. While, the RT-PCR and gene sequencing method were used to detect H5 nucleic acid from environmental swabs from 4 epidemic spots, and hemagglutination inhibition assay was also used to detect H5 antibody.

RESULTSOf 22 environmental swabs detected from 4 epidemic spots, one was positive for H5 nucleic acid, and the homogeneity was 95.9% as compared with H5N1 virus A/China/GD01/2006 (H5N1) found in Guangzhou in 2006 by gene sequence analysis. 62 environmental swabs from live poultry stalls of food markets near epidemic spot were detected negative. Six of 68 blood samples of contacts were positive for H9 antibody, and all were negative for H5 antibody. 68 throat swabs of contacts were detected negative for H5 nucleic acid. No close contact was found abnormal after 7 days medical observation. 337 influenza-like cases were reported in emergency surveillance, and no suspicious case was found.

CONCLUSIONThe current outbreak of H5N1 avian influenza in water fowls has not yet caused further transmission, and human avian influenza case has not been observed. It indicates that the ability of H5N1 virus to transmit to human is not strong yet, and the risk of human infection for H5N1 is still low.

Animals ; Antibodies, Viral ; blood ; China ; epidemiology ; Disease Outbreaks ; Ducks ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; pathogenicity ; Influenza in Birds ; epidemiology ; transmission ; Influenza, Human ; epidemiology ; transmission ; Risk Assessment

Adolescent ; Adult ; Aged ; Child ; Echinococcosis, Hepatic ; complications ; surgery ; Female ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the expression of the human mammoglobin (hMAM) mRNA in bone marrow and its clinical significance in the breast cancer patient.

METHODSExpression of hMAM mRNA was detected using nested reverse transcription polymerase chain reaction (RT-PCR) in the bone marrow aspiration sample from 75 breast cancer patients, 15 patients with benign breast lesions and 8 healthy volunteers as control. The possible correlation of hMAM mRNA expression with clinico-pathological parameters and related molecular markers such as Ki67, p53 and VEGF were analyzed.

RESULTSThe sensitivity of RT-PCR in this series reached 10(-6). The hMAM mRNA was found to be positively expressed by RT-PCR in 21 of 75 breast cancer patients with a positive rate of 28.0%. However, hMAM mRNA expression was not detected in the bone marrow aspiration samples from patients with benign breast lesions and healthy volunteers. The hMAM mRNA expression was positively correlated with axillary nodal involvement and progesterone receptor (PR) status (P < 0.05) as well as Ki67 expression in breast cancer tissue (chi2 = 4.936, P = 0.026), but not with age, tumor size, clinical stage, or estrogen receptor (ER) status (P > 0.05).

CONCLUSIONRT-PCR is quite sensitive and has a high specificity in detecting the presence of hMAM mRNA in the bone marrow from breast cancer patients. Thereupon, hMAM mRNA may be useful as a molecular biomarker in detecting disseminated tumor cells (DTC) in the bone marrow of breast cancer patients. Positive hMAM mRNA expression result may have an impact upon therapeutic recommendations and patients' prognostic judgement.

Adult ; Aged ; Biomarkers, Tumor ; genetics ; Bone Marrow ; metabolism ; pathology ; Breast ; metabolism ; pathology ; Breast Neoplasms ; genetics ; pathology ; Breast Neoplasms, Male ; genetics ; pathology ; Carcinoma, Ductal, Breast ; genetics ; pathology ; Female ; Fibroadenoma ; genetics ; pathology ; Humans ; Ki-67 Antigen ; genetics ; Lymphatic Metastasis ; Male ; Mammaglobin A ; Middle Aged ; Neoplasm Proteins ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Progesterone ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Uteroglobin ; genetics

BACKGROUNDMany studies have shown the superior efficacy of budesonide (BUD)/formoterol (FORM) maintenance and reliever therapy, but still lack evidence of its efficacy in Chinese asthma patients in a relative large patient-group. We finished this research to compare BUD/FORM maintenance and reliever therapy and high-dose salmeterol (SALM)/fluticasone (FP) maintenance plus an as-needed short-acting β(2)-agonist in Chinese patients with persistent uncontrolled asthma. This was a post hoc analysis based on a 6-month, multicenter, randomized, double-blind study (NCT00242775).

METHODSA total of 222 eligible asthma patients from nine centers in China were randomized to either BUD/FORM+as-needed BUD/FORM (160/4.5 µg/inhalation) (640/18 µg/d; n = 111), or SALM/FP+as-needed terbutaline (0.4 mg/inhalation) (100/1000 µg/d; n = 111). The primary endpoint was time to first severe exacerbation while secondary endpoints included various measures of pulmonary function, symptom control and quality-of-life.

RESULTSTime to first severe exacerbation over six months was lower with the BUD/FORM than with the SALM/FP treatment (risk ratio = 0.52, 95%CI 0.22 - 1.22), but the difference did not achieve statistical significance (P = 0.13). The cumulative number of severe exacerbations in the BUD/FORM group was lower than in the SALM/FP group (7.2% vs. 13.5%; risk ratio = 0.45, P = 0.028). BUD/FORM produced significantly better improvements in reliever use, cumulative mild exacerbations, symptom-free days (%), and morning/evening peak expiratory flow (PEF) than SALM/FP (P < 0.05 in all cases). The two groups achieved similar improvements in their time to first mild exacerbation, forced expiratory volume in one second (FEV(1)), asthma control questionnaire and asthma symptom scores, and percentage of nights with awakening(s). Both treatments were well tolerated.

CONCLUSIONSIn Chinese patients with persistent asthma, BUD/FORM decreased severe and mild exacerbations, decreased reliever use, increased symptom-free days, and improved morning/evening PEF compared with SALM/FP. There were no significant differences in time to first severe exacerbation or other assessments regarding daily asthma control between BUD/FORM and SALM/FP. BUD/FORM was more effective in this Chinese sub-group than in the total cohort involved in the original study.

Adolescent ; Adult ; Aged ; Asthma ; complications ; drug therapy ; physiopathology ; Budesonide ; administration & dosage ; adverse effects ; Double-Blind Method ; Ethanolamines ; administration & dosage ; adverse effects ; Female ; Forced Expiratory Volume ; Formoterol Fumarate ; Humans ; Male ; Middle Aged