Porphyromonas gingivalis is a specific causative agent of human chronic periodontitis. This anaerobe produce collagenase PrtC encoded by prtC. To constructed the prokaryotic expression system for the prtC gene from P.gingivalis so as to be used to explore antigenicity and immunoreactivity of prtC gene product as well as the relationship between the local level of PrtC and the periodontitis damage, the entire length of prtC genes fragment from the ATCC-33277 and 47A-1 strains of P.gingivalis was amplified by PCR. After T-A cloning and sequencing, the prokaryotic expression system for prtC was constructed by using pET32a plasmid and E.coli BL21DE3 strain. The expression of the target recombinant PrtC protein was induce with different concentrations of IPTG. Western blot assay was used to detect the antigenicity and immunoreactivity of PrtC protein, and ELISA assay was used to detect the PrtC level in the subgingival plaque samples of patients with chronic periodontitis. The experimental results showed that the nucleotide sequences of the prtC genes from ATCC-33277 and 47A-1 strains of P.gingivalis were entirely identical, and the sequence similarities of nucleotides and amino acids were 98.46% and 99.07% respectively. Under the induction of different concentration of IPTG, the output of recombinant expressed product PrtC may reach up to 50% of the total bacterial proteins. It was also proved that the recombinant PrtC could bind with antibody against the whole cell of P.gingivalis, and could induce the production of specific antibodies in rabbit. In 91.39% of the subgingival plaque samples, the PrtC could be detectable, in which the level of positive rates of detection was higher in severe cases of chronic periodontitis than that in the mild cases. So far, a prokaryotic expression system for the prtC gene from P.gingivalis with high expression efficiency was successfully constructed in the present study, and the expressed product PrtC possesses well antigenicity and immunoreactivity, suggesting the possibility to be used as the candidate antigen for developing the serological kit and P.gingivalis vaccine.

Asia ; Congresses as Topic ; Humans ; Pediatrics

Early Diagnosis ; Early Intervention (Education) ; methods ; trends ; Genetic Diseases, Inborn ; diagnosis ; therapy ; Humans ; Research ; trends

Objective To study the effect of Depside Salt from Salvia Miltiorrhiza on angina pectoris and platelet function.Methods The study group was comprised of 56 patients with stable angina,who were randomly divided into the high-dosage Depside Salt from Salvia Miltiorrhiza group,the low-dosage Depside Salt from Salvia Miltiorrhiza group and the Danshen control group. Before and after the 14-day treatment,the clinical symptom and serum level of PAG, P-selectin were measured,and exercise electrocardiography was performed. Results After treatment with Depside Salt from Salvia Miltiorrhiza,the symptom of angina pectoris was alleviated and exercise ECG was improved,while no significant difference was found compared with control group.Serum PAG and P-selectin were decreased after treatment with Depside Salt from Salvia Miltiorrhiza ,and significantly more than Danshen.Conclusion Depside Salt from Salvia Miltiorrhiza injection can remarkably inhibite the aggregation and activation of platelet,and is effective for angina pectoris.

OBJECTIVETo analyze the change in drug resistance of Staphylococcus aureus (SAU) in the PLA general hospital from January 2008 to December 2012, and to provide solid evidence to support the rational use of antibiotics for clinical applications.

METHODSThe SAU strains isolated from clinical samples in the hospital were collected and subjected to the Kirby-Bauer disk diffusion test. The results were assessed based on the 2002 American National Committee for Clinical Laboratory Standards (NCCLS) guidelines.

RESULTSSAU strains were mainly isolated from sputum, urine, blood and wound excreta and distributed in penology, neurology wards, orthopedics and surgery ICU wards. Except for glycopeptide drugs, methicillin-resistant Staphylococcus aureus (MRSA) had a higher drug resistance rate than those of the other drugs and had significantly more resistance than methicillin-sensitive Staphylococcus aureus (MSSA) (P < 0.05). In the dynamic observation of drug resistance, we discovered a gradual increase in drug resistance to fourteen test drugs during the last five years.

CONCLUSIONDrug resistance rate of SAU stayed at a higher level over the last five years; moreover, the detection ratio of MRSA keeps rising year by year. It is crucial for physicians to use antibiotics rationally and monitor the change in drug resistance in a dynamic way.

Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Multiple, Bacterial ; Humans ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; Staphylococcal Infections ; drug therapy ; Staphylococcus aureus ; drug effects

Child ; Humans ; Puberty, Precocious ; diagnosis ; therapy

Adult ; Aged ; Aged, 80 and over ; Female ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; classification ; pathogenicity ; Humans ; Lymphatic Metastasis ; Male ; Microvessels ; pathology ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; microbiology ; pathology ; Stomach Neoplasms ; blood supply ; metabolism ; microbiology ; Vascular Endothelial Growth Factors ; metabolism

Animals ; Calcium ; analysis ; physiology ; Cells, Cultured ; Motilin ; pharmacology ; Myocytes, Smooth Muscle ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley

A novel nucleic acid amplification method,termed loop-mediated isothermal amplification(LAMP),which amplifies DNA with high specificity,efficiency,and rapidity under isothermal conditions,may be a valuable tool for the rapid detection of infectious diseases.This method employs a DNA polymerase that have activity of strand displacement DNA synthesis and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.LAMP can amplify a few copies of DNA to 109 in less than an hour.The final products are stem-loop DNA with several inverted repeats of the target and cauliflower-like structures with multiple loops.A positive reaction would be shown as a ladder-like pattern in a gel electrophoresis analysis.Because of the advantage,the LAMP method will be widely applied to research of nucleic acid,clinical diagnosis of infectious diseases and detection of genetically modified organisms etc.

Body Height ; drug effects ; Child ; Drug Monitoring ; Growth Disorders ; diagnosis ; drug therapy ; Human Growth Hormone ; administration & dosage ; adverse effects ; therapeutic use ; Humans ; Practice Guidelines as Topic ; Recombinant Proteins ; administration & dosage ; adverse effects ; therapeutic use ; Risk Factors