Objective To study the AmpC beta-lactarnase and its genotype mediated by plasmid in Escherichia coli.Methods AmpC beta-lactamase was detected based on that AmpC beta-laetamase can be inhibited by 3-aminophenylboronic acid(APB).MIC was ehecked by agar dilution method.Conjugation test was used to check the transfer of ampC gene.Gene chip and PCR were used to detect ampC gene.The amplified ampC gene were sequenced and analyzed by EMBOSS software.The molecular epidemiology of clinical isolates was investigated by Enterbacterial repetitive intergenic consensus(ERIC)typing method.Results In 74 strains of Escherichia coli insusceptible to cefoxtin,AmpC beta-lactamase was positive in 33 strains.8 strains possessed AmpC beta-lactamase of CIT group by gene chip and 8 transconjugants were obtained by conjugation test.CMY type ampC gene could be further amplified by specific CMY gene primers from both 8 clinical isolates of E.coli and plasmids extracted from 8 transconjugants.CMY-2 type ampC gene was found by sequencing(accession number DQ823449).The most transconjugants displayed similar MIC value(intermediate or resistant).ERIC genotyping showed 6 out of 8 isolates with CMY-2 ampC gene derived from different resource.Conclusion CMY-2 AmpC beta- lactamase mediated by plasmid could be detected in E.coli isolates from patients in the General Hospital of People Liberation Army,Beijing.The plasmid carried ampC gene could mediate multi-drug resistance.

Objective To investigate the molecular epidemiological characteristics of Staphylococcus aureus associated with bloodstream infection in hospital. Methods 47 Staphylococcus aureus strains isolated from bloodstream in PLA General Hospital were collected from January 2006 to December 2008. Susceptibility of the strains to 11 antimicrobial agents was detected and DNA homology of them was analyzed with Rep-based DiversiLab~(TM) Microbial Typing System. Panton-Valentine leukocidin (PVL)gene was determined by PCR. For methicillin-resistant Staphylococcus aureus (MRSA) strains,the genotypes of SCCmec were determined and ST239 clone was screened with multiplex PCR. Multilocus sequence typing (MLST) was used to determine the STs of the selected isolates. Results In the 47 Staphylococcus aureus isolated from blood,methicillin-resistant strains accounted for 51.1%,all belonged to SCCmec Ⅲ type,with only 2 pvl gene positive strains identified. 12 different patterns (A-L) were found among 47 strains with Rep-PCR. All MRSA strains clustered in the A and B subtypes. Conclusion Most MRSA strains isolated from blood in PLA General Hospital belonged to ST239-MRSA-SCCmec Ⅲ clone. DiversiLab~(TM) Microbial Typing System could provide a rapid and effective method to investigate the molecular epidemiological characteristics of Staphylococcus aureus in the hospital settings.

Objecfive To investigate antibiotic resistance,carbapenemase genotype and the molecular epidemiology of multidrug-resistant Acinetobacter baumannii (Aba) collected from 3 military hospitals in China.Methotis The minimum inhibitory concentrations (MIC) were examined by ager dilution method.Genotypes of carbapenemases were amplified by multiplex PCR and its products were sequenced.PCR was used to detect per gene,Homology of the resistant isolates was analyzed by pulse-field gel electrophoresis(PFGE).Results Among the 64 MDRA strains,78.1%(50)strains possessed blaOXA-23 gene,89.1%(57) carried Class 1 integrase gene,39.1% (25) with blaPER-1 gene,and 1 strain with blaOXA-58-like gene.PFGE showed that 13(A,B,C,D,E genotype) different clones were identified in these strains.A,B,and U clones were the predominant clones in three hospitals,respectitively.Conclusion OutbreaEs of muitidrug-rcsistant Aba occurred at 3 military hospitals with the most prevalent carbapenemase as OXA-23 enzyme.OXA-58 type of carbapenemase and per-1 in Aba were also isolated.

Objective To investigate the efficacy and feasibility of preimplantation genetic diagnosis (PGD) with human leukocyte antigen (HLA) matching for beta-thalassemia. Methods A total of 43 referred beta-thalassemia couples, with at least on child in need of hematopoietic stem cell transplantation (HSCT), underwent PGD for HLA matching at the First Affiliated Hospital of Sun Yat-sen University from 2010 to 2015. PGD cycles of these couples were retrospectively analyzed, and 15 infants born from PGD-HLA were followed up. Results A total of 84 oocyte retrieval cycles were performed, providing 14±7 oocytes per cycle. Fifty nine embryos biopsied cycles were done, including 24 cleavage stage and 35 blastocyst stage biopsy cycles. In cleavage stage, 259 embryos were biopsied, 93.4% (242/259) of them with conclusive molecular diagnosis, and the percentage of unaffected embryos (normo-homozygote and heterozygote) was 71.4%(185/259). The percentage of HLA matched unaffected embryos was 9.3%(24/259). In blastocyst stage, 306 embryos were biopsied, while 93.8% (287/306) of them were conclusive, and the percentage of unaffected embryos was 70.6% (216/306). The percentage of HLA matched unaffected embryos in blastocyst stage biopsy was 14.4%(44/306), which was higher than in cleavage stage biopsy (P<0.05). Forty three female carriers underwent 48 embryo transfer cycles including 3 fresh and 45 frozen-thawed embryo transfer cycles. Three fresh embryo transfer cycles were done after cleavage stage biopsy, resulted in a birth of healthy twins born at 36 weeks′gestation. All the embryos were frozen after blastocyst biopsied. Totally, 54 frozen-thawed embryos that were transferred in 45 frozen-thawed embryo transfer cycles included 25 embryo from cleavage stage biopsy and 29 embryo from blastocyst stage biopsy, and 42 of them were HLA matched. Clinical pregnancy rate and implantation rate per cycle in frozen-thawed embryo transfer were 38%(17/45) and 37%(20/54) respectively. A total of 15 infants were born, 2 were from a fresh embryo transfer cycle, and 13 were from frozen-thawed embryo transfer cycles. Results of prenatal diagnosis from delivered cases were matched to that of PGD. Four sick children have been cured by HSCT from these HLA matched born siblings. Conclusion PGD with HLA matching is an established method for conceiving a child who may donate hematopoietic stem cells to save an ill sibling.

OBJECTIVETo study the mode of transmission and molecular characteristics on carbapenem-resistant Acinetobacter baumannii strain. Strains were isolated from different parts of samples in various patients.

METHODSClinical information of carbapenem-resistant Acinetobacter baumannii isolates were stored and analyzed by WHONET 5.4 software. The transmission and pathopoiesis of the strains were learned through case file review. Genotypes of isolates were identified by pulse-field gel electrophoresis (PFGE) and genes of carbapenemase were detected by multiple PCR, in order to find molecular characteristics and relatedness between strains.

RESULTS29 stains of Acinetobacter baumannii resistant to carbapenem were isolated from 2 or more kinds of samples among 13 patients'. Two genotypes were identified by PFGE: genotype A was obtained from 22 isolates in 11 patients and genotype B was obtained from 7 isolates in 4 patients. PCR amplification showed that all strains possessed OXA-23 gene except 1, and all strains possessed Integrase gene I except 3.

CONCLUSIONThere were 2 different genotypes from 29 strains of carbapenem-resistant Acinetobacter baumannii with Genotype A as the main type. OXA-23 carbapenemase gene and integrase gene I were detected from most of the isolates. All the strains could be easily transmitted in the body of the patients and among patients, hence becoming the epidemic pathogen of iatrogenic infection.

Acinetobacter Infections ; microbiology ; transmission ; Acinetobacter baumannii ; classification ; drug effects ; genetics ; Carbapenems ; pharmacology ; Cross Infection ; microbiology ; transmission ; Drug Resistance, Bacterial ; genetics ; Genotype ; Humans

OBJECTIVETo investigate the effect of rhein on the development of hepatic fibrosis.

METHODSThe animal models were made with carbon tetrachloride (CCl(4)) mixed with vegetable oil (3/2, v/v), which was injected subcutaneously twice a week for 6 weeks, and with 5% ethanol for free drinking water. At the same time, Rhein was administrated at the dose of 25 mg/kg or 100 mg/kg once a day for 6 weeks. The changes of both biochemical markers, such as the levels of alanine aminotransferase (ALT), hyaluronic acid (HA), procollagen type III (PCIII) in serum and SOD, malondialdehyde (MDA) in liver, and related histopathological parametres were determined.

RESULTSCompared with the model group, there were three kinds of changes in the larger quantity of rhein treated group. (1) The levels of ALT, HA, PCIII in serum and MDA in liver homogenate were decreased significantly (from 150 U/L +/- 16 U/L to 78 U/L +/- 18 U/L, 321 microg/L +/- 97 microg/L to 217 microg/L +/- 75 microg/L, 31 microg/L +/- 14 microg/L to 16 microg/L +/- 6 microg/L and 3.67 nmol/mg +/- 0.68 nmol/mg to 1.88 nmol/mg +/- 0.34 nmol/mg, respectively, t > or 2.977, P<0.01). However the level of SOD in liver was increased (from 62.45 NU/mg +/- 8.74 NU/mg to 91.26 NU/mg +/- 14.04 NU/mg, t=4.453, P<0.01). (2) The expressions of transforming growth factor beta 1 (TGF-beta 1) and alpha-smooth muscle actin (alpha-SMA) in liver were markedly reduced (P<0.05 and P<0.01). (3) The collagen staining positive area was decreased and the grade of fibrosis was reduced significantly in liver (P<0.05 and P<0.01).

CONCLUSIONRhein can protect hepatocyte from injury and prevent the progress of hepatic fibrosis in rats, which may associate with that rhein plays a role in antioxidation, anti-inflammation, inhibiting the expression of TGF-beta1 and suppressing the activation of hepatic stellate cells (HSCs).

Animals ; Anthraquinones ; pharmacology ; therapeutic use ; Anti-Inflammatory Agents ; pharmacology ; Antioxidants ; pharmacology ; Collagen ; analysis ; Liver ; drug effects ; pathology ; Liver Cirrhosis, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Rats ; Rats, Wistar ; Transforming Growth Factor beta ; antagonists & inhibitors ; Transforming Growth Factor beta1

OBJECTIVETo investigate the antithrombotic mechanisms of holothurian glycosaminoglycan (GAG) extracted from sea cucumber.

METHODSHuman endothelial cell line EA. hy926 cells were treated with 10 mg/L GAG or 10U/mL unfractionated heparin (UFH) by short-term (15 min - 2 h) and longer-time incubation (6 h - 48 h). Different doses of GAG were used to stimulate EA. hy926. Released free tissue factor pathway inhibitor(TFPI) was determined by ELISA assay. TFPI expression was investigated by immunofluorescent method and TFPI mRNA level by real-time PCR. In a 96-wells microtitre plate, pooled normal plasma containing different concentrations of GAG was allowed to clot by addition of thrombin and calcium chloride, fibrinolysis was induced by addition of t-PA. TRR (TAFI-related retardation of clot lysis) was used to assess thrombin-activatable fibrinolysis inhibitor(TAFI) functional activity.

RESULTSGAG increased TFPI synthesis, expression and secretion in a dose- and time dependent manner. GAG at low concentrations could lengthen while at intermediate concentrations could shorten clot lysis times significantly as compared to control values. TRR was dose-dependently decreased on addition of GAG.

CONCLUSIONSGAG increases TFPI synthesis, expression and secretion of endothelial cells. GAG at intermediate concentrations significantly affects clot stability of a developing clot by means of diminishing TAFI activation.

Animals ; Carboxypeptidase B2 ; antagonists & inhibitors ; Cell Line ; Dose-Response Relationship, Drug ; Endothelial Cells ; drug effects ; metabolism ; Glycosaminoglycans ; pharmacology ; Heparin ; pharmacology ; Holothuria ; Humans ; Lipoproteins ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; Tissue Extracts ; pharmacology

The advent of the Post-Human Genome Project Era, represented by the raising of proteomics, would inevitably lead to the change of molecular biology from topical view to holistic with its thought turning from linear to complex mode. Based on the proteomics development in recent years, the authors summarized the methodology of TCM syndromatologic research, advocated in using two-dimensional gel electrophoresis (2-DE) and bioinformatics to identify different proteins. Proteomics should be led into the research of TCM syndrome categorization and the rule of evolution, which is necessary for researching the integration of the TCM study with proteomics and even with modern molecular biology based on molecular epidemiology level. Owing to the gradually developed coherence and mutual penetration of proteomics and TCM on the thinking method in studying life science, it has denoted the necessity and importance of integration of TCM and Western medicine in investigating the complex vital life phenomena.
Diagnosis, Differential ; Drug Therapy ; methods ; Humans ; Medicine, Chinese Traditional ; methods ; Proteomics ; methods

AIMTo study the pharmacokinetics of fudosteine in healthy volunteers after the single and multiple dose administration.

METHODSThirty-six volunteers were divided into three groups randomly, each group included six men and six women. In the single dose design, the volunteers received either a single dose of 600 mg, 400 mg or 200 mg fudosteine. After a one-week wash out period, the volunteers of 400 mg group participated in the multiple dose design in which each volunteer received 400 mg fudosteine three times a day for five consecutive days. The plasma concentrations were determined by pre-column derivatization HPLC-FL method and the pharmacokinetic parameters of fudosteine were calculated.

RESULTSThe obtained pharmacokinetic parameters of fudosteine in single dose of 600 mg, 400 mg and 200 mg groups were as follows: T1/2 were (2.8 +/- 0.5), (2.7 +/- 0.5) and (3.2 +/- 0.6) h, respectively. T(max) were (0.51 +/- 0.22), (0.59 +/- 0.21) and (0.48 +/- 0.18) h, respectively. C(max) were (16 +/- 4), (11 +/- 3) and (6.1 +/- 1.5) microg x mL(-1), respectively. The AUC(0-10 h) and C(max) correlated linearly with doses, respectively (r > 0.99). The T(max), C(max) and AUC values of fudosteine in healthy male volunteers were smaller than those in female volunteers, and the T1/2 value was longer than that in female volunteers. The obtained multi-dose pharmacokinetic parameters of fudosteine were as follows: C(ss) was (4.1 +/- 0.8) microg x mL(-1); DF was 3.0 +/- 0.7; T1/2 was (2.5 +/- 0.4) h; T(max) was (0.6 +/- 0.3) h; C(max) was (13.2 +/- 1.3) microg x mL(-1).

CONCLUSIONThe values of pharmacokinetic parameters in healthy volunteers were linear in the range from 200 mg to 600 mg. Statistic analysis results showed that the differences of AUC and C(max) between men and women were not resulted from sexual differences, but from the weight differences. There was no significant difference in pharmacokinetic parameters between single dose and multi-dose.

Administration, Oral ; Adult ; Area Under Curve ; Body Weight ; Chromatography, High Pressure Liquid ; methods ; Cystine ; administration & dosage ; analogs & derivatives ; pharmacokinetics ; Dose-Response Relationship, Drug ; Female ; Humans ; Male ; Sex Factors

OBJECTIVETo investigate the sleep status of college and high schools students.

METHODSPittsburgh sleep quality index (PSQI) and self-manufactured questionnaires about siesta habits were used as tools. Three groups of students from medical college (MC), senior high school (SS) and junior high school (JS) were surveyed.

RESULTSIn the group MC, SS and JS, the occurrence rates of sleep disorders were 27%, 62% and 54%, respectively, and in which the appearance rates of insomnia were 17%, 19% and 19%, longing for sleep were 10%, 43% and 35% respectively. And there were no significant difference between schoolboy and schoolgirl. The occurrence rates of slack breathing were different (5/155, 1/154) significantly between group SS and JS. The distinct differences also were found in group JS and MC, in which students felt hot (10/155, 1/122) and in all the three groups, in which students felt sleepy (55/155, 62/154, 13/122) whereas the difference of sleepy between group SS and JS was comparatively distinct (55/155, 62/154). Significant differences were also found between group JS and SS, MC in average sleep time of (7.65 +/- 0.87) hours, (7.16 +/- 0.83) hours, and (7.10 +/- 0.57) hours. The time of falling asleep (median 15 min, 10 min, 20 min) and siesta habit (8/155, 19/154, 75/122) among group MC and SS, JS were different respectively and markedly, whereas siesta habit differences between group SS and JS were comparatively distinct (8/155, 19/154).

CONCLUSIONStudents in high school showed higher rate of longing for sleep, and this implicated they fall short of sleep time greatly and siesta could improve their sleepy signs.

Adolescent ; Adult ; China ; epidemiology ; Female ; Humans ; Male ; Prevalence ; Sleep ; physiology ; Sleep Initiation and Maintenance Disorders ; epidemiology ; Sleep Wake Disorders ; epidemiology ; Students ; Surveys and Questionnaires