Objective To investigate status of macrolide resistance and determine molecular mechanisms in Mycoplasma pneumoniae.Nethods All of 370 throat swab specimens were cultured to isolate Mycoplasma pneumoniae.Mycoplasma pneumoniae isolates were identified by nested PCR for specific 16SrRNA gene.Antibiotic susceptibility test was done to identify acrolide resistant strains.23SrRNA gene wag amplified by nested PCR followed by direct automatic sequencing method.The DNA sequences were compared to the sequence of Mycoplasma pneumoniae M129(accession no.X68422)to find molecular mechanisms of drug resistance.Results Fifty clinical strains were isolated from 370 specimens.Of 50 strains.4 strains were susceptible to macrulide,46 strains were macrolide resistant with the percentage of 92%.MICs of resistant strains to erythromycin.Azithromycin and josamycin were elevated.The sequence of 23SrRNA gene in 4 Susceptible strains and the reference strain FH was identical to Mycoplagma pneumoniae gene in GenBank.46 resistant strains arbored a point mutation respectively,among them,40 strains had all A to G transition at position 2063.1 strain had an A to C transition at position 2063,the other five strains showed an A to G transition at position 2064.Conclusions Macrolide resistance in Mycoplasma pneumoniae iS very serious health conceru.The point mutation in 23SrRNA.Xpecailly predominant position 2063 mutation contributed to the macrolide resistance in Mycoplagma pneumoniae.The MICs of resistant strains to erythromycin,azithromycin and iosamycin are much higher than Mycoplasma pneumoniae reference strain FH.

Chemical and Drug Induced Liver Injury ; Hair Dyes ; toxicity ; Humans ; Liver ; drug effects ; pathology ; Liver Diseases ; diagnosis ; drug therapy ; etiology ; Male ; Young Adult

OBJECTIVETo investigate the effects of mTOR siRNA on mTOR-p70S6K signaling pathway in esophageal squamous cell carcinoma (ESCC) cells in vitro,and growth and apoptosis in transplanted tumor in nude mice.

METHODSmTOR siRNA was transfected into ESCC cell line EC9706 cells. The expressions of factors of the mTOR/p70S6K signaling pathway were detected by RT-PCR and Western blot. DNA contents and cell apoptosis were determined by flow cytometry, and cell proliferation was measured by CCK-8 assay. The effects of mTOR siRNA on the transplanted tumor growth were assessed in nude mice.

RESULTSThe levels of mTOR and p-p70S6K were significantly decreased (P < 0.05) while the level of p70S6K was increased (P < 0.05) in the cells transfected with mTOR siRNA, compared with that in untransfected cells and cells transfected with control siRNA. After being interfered by mTOR siRNA, the number of apoptotic cells was increased, cell proliferation became slower and cell cycle was arrested in G(1) phase compared with that in control cells. Also, mTOR siRNA inhibited the growth of transplanted tumor in vivo.

CONCLUSIONSmTOR siRNA can effectively interfere in mTOR-p70S6K signaling pathway, induce cell apoptosis and inhibit cell proliferation and tumor growth, suggesting that mTOR-p70S6K signaling pathway plays an important role in the carcinogenesis and development of esophageal squamous cell carcinoma.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; enzymology ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; enzymology ; pathology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; pharmacology ; Ribosomal Protein S6 Kinases, 70-kDa ; metabolism ; Signal Transduction ; TOR Serine-Threonine Kinases ; genetics ; metabolism ; Transfection ; Tumor Burden

One pair of degenerate primer was designed according to conserved motifs of the psaB (A2 subunit of photosystem I) of Chlamydomonas reinhardtii, Chlamydomonas moewusii, Chlorella vulgaris and Mesostigma viride, and a total RNA of Dunaliella salina (D. salina) was extracted with TRIzol reagent. A cDNA fragment, about 1.8kb in length, from green algal D. salina was obtained through RT-PCR method. The resulting PCR product was cloned into T-vector and screened to determine its sequence. Homologous analysis of the deduced amino acid sequence was performed by BLAST and subsequeqtly compared with GenBank data. The obtained cDNA sequence was 1815 bp long, which encodes 605 amino acids (GenBank accession number: AY820754). The sequence shared high homologue with the following psaB: Chlamydomonas reinhardtii 92%, Chlamydomonas moewusii 91%, Chlorella vulgaris 86%, Mesostigma viride 85%, Physcomitrella patens subsp. Patens 85% and Nephroselmis olivacea 84%. It can be concluded that the cloned sequence is psaB cDNA fragment from D. salina.
Algal Proteins ; genetics ; Amino Acid Sequence ; Animals ; Chlamydomonas reinhardtii ; genetics ; Chlorophyta ; genetics ; metabolism ; Cloning, Molecular ; DNA, Complementary ; genetics ; Molecular Sequence Data ; Photosystem I Protein Complex ; genetics ; Sequence Analysis, Protein ; Sequence Homology, Amino Acid

OBJECTIVETo study the risk factors of male infertility.

METHODCase control study including 94 cases and control group with a ratio of 1 to 1.

RESULTSThe risk factors of male infertility were long time heavy smoking habit (OR = 3.45, 95% CI: 1.95 - 6.10), illegal sexual intercourse (OR = 7.29, 95% CI: 2.54 - 20.89), growing vegetable under plastic in higher temperature (OR = 6.73, 95% CI: 1.91 - 23.69), contact with benzene chemicals (OR = 20.53, 95% CI: 4.67 - 90.25) and having Ureaplasma urealyticum (Uu) infection (OR = 5.55, 95% CI: 2.28 - 13.53).

CONCLUSIONMale infertility was resulted from many factors repeatedly acting on men for long time. In order to prevent male infertility, issues as environmental pollution, occupational protection need to be improved while bad working condition and risky behavior should be changed.

Adult ; Benzene ; toxicity ; Case-Control Studies ; China ; epidemiology ; Environmental Pollution ; adverse effects ; Humans ; Infertility, Male ; epidemiology ; etiology ; Male ; Occupational Exposure ; adverse effects ; Risk Factors ; Smoking ; adverse effects ; Ureaplasma Infections ; complications ; Ureaplasma urealyticum

OBJECTIVETo study the molecular mechanism of TAp63gamma-induced cell apoptosis.

METHODSTranscription and protein expression of apoptosis inducing factor and p63 were investigated by immunohistochemistry and RT-PCR in human esophageal squamous carcinoma cell line EC9706 respectively. Twenty-four hours after transfection with pcDNA3.1-TAp63gamma, the apoptosis and translocation of apoptosis inducing factor in EC9706 cells were studied by flow cytometry, laser confocal microscopy and mitochondrial/cytosol/nuclear extraction analysis respectively. Down-regulation of apoptosis inducing factor protein was achieved by RNAi and pretreatment with caspase inhibitor zVAD.fmk of EC9706 cells.

RESULTSPresence of protein expressions of apoptosis inducing factor and absence of TAp63gamma was observed in the cytoplasm of untransfected cells. RT-PCR verified the subtype of p63 in EC9706 cells was DeltaNp63. After 24 hours of transfection, both nuclear and cytoplasmic expression of apoptosis inducing factor protein were observed in cells transfected with TAp63gamma and p53 expression vectors, but not in cells transfected with control vector. Cell apoptosis rates were 1.37%, 13.64%, 4.52%, 4.03% and 1.91% in the pcDNA3.1 transfection group, pcDNA3.1-TAp63gamma transfection group, apoptosis inducing factor siRNA and pcDNA3.1-TAp63gamma transfection group, zVAD.fmk treatment group, and the group receiving apoptosis inducing factor siRNA, plus zVAD.fmk treatment and pcDNA3.1-TAp63gamma transfection, respectively.

CONCLUSIONSApoptosis inducing factor of EC9706 cells is released from mitochondria into both the cytoplasm and nucleus during TAp63gamma induced apoptosis. Down-regulation of apoptosis inducing factor inhibits TAp63gamma-induced apoptosis. Overall, TAp63gamma-induced apoptosis is dependent on the expression of apoptosis inducing factor and caspase.

Amino Acid Chloromethyl Ketones ; pharmacology ; Apoptosis ; Apoptosis Inducing Factor ; genetics ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Caspase Inhibitors ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Down-Regulation ; Esophageal Neoplasms ; metabolism ; pathology ; Humans ; Mitochondria ; metabolism ; Plasmids ; Protein Transport ; RNA Interference ; RNA, Small Interfering ; genetics ; Trans-Activators ; genetics ; metabolism ; Transcription Factors ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo explore the association of metabolic syndrome with CVD in a cohort study involving 11 provinces.

METHODSA cohort study was carried out in a population of 27 739 subjects (age 35 - 64 years). A baseline survey on the risk factor of cardiovascular disease was done in 1992. Incidence rate and relative risk were calculated for metabolic syndromes.

RESULTS(1) The age-standardized incidence rate of cardiovascular disease for metabolic syndromes (MS) was higher than those without (MS) in this cohort (MS 652.3/100,000, not MS 206.7/100,000, RR = 3.12, P < 0.001). (2) The incidence rate of cardiovascular disease for patients with hypertension but low HDL cholesterol and high waist was the highest (910.2/100,000 in man, 930.7/100,000 in woman) among all metabolic syndromes patients. (3) The predictive risk factors for cardiovascular disease were age, cigarette smoking, BMI, total cholesterol and metabolic syndrome in men, with age, BMI and metabolic syndrome in women.

CONCLUSIONThe incidence rate of cardiovascular disease on the metabolic syndrome was high in 11 provinces in China. Age, BMI and metabolic syndrome were the predictive risk factors of cardiovascular disease (especially of cerebrovascular disease). It is essential to prevent risk factors of cardiovascular disease in primary and secondary prevention programs in general population.

Adult ; Body Mass Index ; China ; epidemiology ; Cohort Studies ; Coronary Disease ; epidemiology ; etiology ; Female ; Humans ; Incidence ; Male ; Metabolic Syndrome ; complications ; epidemiology ; Middle Aged ; Proportional Hazards Models ; Risk Factors ; Smoking ; adverse effects ; Stroke ; epidemiology ; etiology

OBJECTIVETo establish a quick method to detect drug resistance of Mycoplasma pneumoniae (MP) and study the condition of drug resistance in MP infection.

METHODSMP 23S rRNA target gene in throat swab specimens from 200 patients with suspected MP infection was detected by using nested PCR and DNA sequencing. The result of 23S rRNA gene detection was confirmed by MP isolation and drug susceptibility test in vitro for reliability.

RESULTSOf the 200 clinical specimens, 64 were proved to be positive for MP through MP-IgM antibody, MP specific 16S rRNA nested PCR and MP isolation . The 23S rRNA gene was amplified and the gene sequence was compared with MP reference strain in Genbank, 26 were identical to the reference strain, 38 had a point mutation in 23S rRNA. Among them, 35 had A to G mutation at position 2063, 1 had A to C mutation at position 2063 and 2 had A to G mutation at position 2064, the percentage of drug resistance was 59.4%. The sensitivity of the gene detection method was 10(2) ccu/ml and it was confirmed to be reliable by MP isolation and drug susceptibility test.

CONCLUSIONSThe gene detection method could detect MP drug resistant gene directly from clinical specimen, which has the advantages of high specificity, high sensitivity and quickness. It is of great significance for diagnosis of MP infection because MP isolation is difficult and time-consuming.

Adolescent ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Genes, rRNA ; Humans ; Infant ; Male ; Microbial Sensitivity Tests ; Mycoplasma pneumoniae ; genetics ; isolation & purification ; Point Mutation ; Polymerase Chain Reaction ; RNA, Bacterial ; genetics ; RNA, Ribosomal, 23S ; genetics

OBJECTIVETo evaluate the clinical efficacy and safety of integrative medical program based on blood cooling and detoxification recipe (BCDR) in treating patients with hepatitis B virus related acute-on-chronic liver failure (HBV-ACLF) of heat-toxicity accumulation syndrome (HTAS).

METHODSAdopting randomized controlled clinical design, a total of 105 HBV-ACLF patients of HTAS were randomly assigned to the trial group (64 cases) and the control group (41 cases). Patients in the control group were treated with comprehensive Western therapy, while those in the trial group were treated with comprehensive Western therapy plus BCDR. All were treated for 8 weeks and followed up for 40 weeks. Effect and safety of the treatment were assessed, including fatality, liver functions [total bilirubin (TBIL), albumin (ALB), alanine aminotransferase (ALT), and aspartate transaminase (AST)], and prothrombin activity (PTA) after treatment and at week 48 of follow-ups.

RESULTSAfter 8-week treatment, there was statistical difference in the overall fatality rate (15.63% vs 34.15%), the fatality rate in the mid-term (25.0% vs 64.7%), TBIL at week 8 (64.54 +/- 79.75), AST [at week 2: (178.97 +/- 44.24) U/L vs (288.48 +/- 58.49) U/L; at week 4: (61.65 +/- 27.36) U/L vs (171.12 +/- 89.11) U/L] and PTA [at week 4: (58.30 +/- 15.29) vs (42.56 +/- 15.27); at week 6: (60.77 +/- 20.40) vs (43.08 +/- 12.79)] (all P < 0.05). At week 48 of the followup, the fatality rate of the trial group (21.88%) decreased by 17. 14% when compared with that of the control group (39.02%; P < 0.05). No obvious adverse event occurred in the two groups during the 8-week treatment period.

CONCLUSIONBCDR could significantly reduce the mortality of HBV-ACLF patients.

Acute-On-Chronic Liver Failure ; drug therapy ; virology ; Adult ; Drugs, Chinese Herbal ; therapeutic use ; End Stage Liver Disease ; Female ; Hepatitis B virus ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Phytotherapy ; Young Adult

OBJECTIVETo evaluate the correlation between the efficacy of interferon-alpha-2a and the kinetics of viral load in serum.

METHODSThe authors conducted a trial including 58 patients with chronic hepatitis B. Patients were treated with interferon-alpha-2a three times a week for 6 months. Viral kinetics were assessed by serial quantitive measurements of HBV-DNA.

RESULTSA significant decline of serum HBV-DNA was seen after interferon-alpha-2a administration for 1 month, the decreases were (2.50 +/- 0.44) log10, (1.62 +/- 1.12) log10 and (1.05 +/- 1.35) log10 for complete responders, partial responders and no-responders, respectively. After 1 month of treatment, HBV-DNA level was (3.99 +/- 0.91) log10 for complete responders versus (5.63 +/- 1.31) log10 for partial responders, and (6.69 +/- 1.42) log10 for no-responders (P < 0.05). Multivariate analysis suggested that undetectable serum HBV-DNA after 1 month of interferon-alpha-2a treatment was associated with better efficacy; higher baseline ALT or/and no family history were also correlated with better treatment outcomes.

CONCLUSIONKinetics of HBV-DNA level under interferon-alpha-2a treatment are highly predictive of therapeutic response.

Antiviral Agents ; therapeutic use ; CD13 Antigens ; blood ; China ; DNA, Viral ; blood ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Multivariate Analysis ; Polymerase Chain Reaction ; Treatment Outcome