Objective To obtain the recombinant protein LytA (rLytA) of Streptococcus pneumoniae strain (ATCC49619) through prokaryotic expression system and to investigate their diagnostic value for patients with community acquired pneumonia (CAP).Methods The specific primers were designed according to LytA gene sequence of Streptococcus pneumoniae M66 strain recorded in Genbank.The recombinant plasmid pET32a(+)/LytA was constructed and transformed into BL21(DE3) to express LytA.The expressed protein LytA was purified by electroeluting of bag filter.Serum IgM of anti-LytA accordingly of patients with CAP were detected by ELISA.The results were evaluated by Chi-square test.Results The recombinant protein LytA was expressed and purified successfully with a relative molecular weight of 56 000.The IgM antibodies level of anti-LytA was significantly higher than the healthy control group (P=0.000).Diagnostic sensibility and specificity of LytA-IgM were 27.8% and 100.0%,while sensibility and specificity of sputum culture were 19.4% and 72.2%,respectively.The sensibility of LytA-IgM was equal to sputum culture(χ2=0.693,P=0.405),but the specificity was higher than it(χ2=14.316 P=0.000).Conclusions A rLytA-ELISA assay maybe has clinical value for diagnosis of pneumococcal infections.It is more rapid and objective than the culture method.

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

Objective To investigate the reliability of the Three-Column classification for fracture of tibial plateau and compared with the conventional Schatzker classification.Methods From December 2004 to March 2007,304 patients with tibial plateau fracture were treated in our department.The CT scans and 3D reconstruction were performed in all patients before operation.All the patients were classified by the Three-Column classification and then treated with open reduction and internal fixation via the corresponding approaches based on the Three-Column classification.To validate the Three-Column classification,four doctors classified 50 patients which were chosen randomly from 323 consecutive fracture cases.Schatzker classification was also classified at the same time.Inter-observer reliabilities of the Three-Column classification and the Schatzker classification were figured out respectively in the form of Kappa value.Results According to the Three-Column classification,there were 4 pure compression fractures (1.2％),181 one-column fractures(56.0％),108 two-column fractures(33.4％) and 30 three-column fractures (9.3％) in all 323 cases.In one-column fracture cases,the numbers of lateral,medial,and posterior column fractures were 114 (63.0％),53 (29.3％),and 14 (7.7％),respectively.Among two-column fracture cases,the lateral-medial column fractures (33.3％) were 36.While the number of medial-posterior column fractures and lateral-posterior column fractures were 34 (31.5％) and 38 (35.2％),respectively.There was a moderate inter-observer reliability when using Schatzker classification (Kappa=0.567; range,0.513-0.589).There was a substantial interobserver reliability when using the Three-Column classification (Kappa=0.766; range,0.706-0.890).Conclusion The Three-Column classification based on CT and 3D reconstruction is simple and more reliable for fracture of tibial plateau.It is suggested that the new classification system,the Three-Column classification,can be used and popularized in the orthopaedic surgery.

Objective To construct the recombinant protein AtlM ( rAtlM) of Staphylococcal au-reus through prokaryotic expression system and to investigate its antibacterial activity in vitro.Methods The specific primers were designed according to atlM gene sequence of Staphylococcal aureus recorded in Gen-Bank, and atlM gene was amplified by PCR from the Staphylococcal aureus strain (ATCC25923).The re-combinant plasmid pET-32а(+)/atlM was constructed and transformed into Transetta ( DE3 ) to express AtlM after induced by IPTG .The expressed protein AtlM was further analyzed by SDS-PAGE and purified by electroeluting of bag filter.The minimal inhibitory concentrations (MICs) of rAtlM to ATCC25923 and oxac-illin-resistant S.aureus strain were determined by the broth microdilution method .S.aureus ATCC25923 strain and oxacillin-resistant S.aureus strain (final concentration of 5×105 CFU/ml) were exposed to rAtlM (50 μg/ml) respectively to test its antibacterial activity in vitro.Results The recombinant protein AtlM was expressed and purified successfully with a relative molecular weight of 80 ×103 and a concentration of 1.25 mg/L.The MICs of rAtlM to ATCC25923 strain and oxacillin-resistant S.aureus strain were 8 μg/ml and 64 μg/ml, respectively.In vitro test showed that rAtlM had inhibitory effects on the growth of ATCC25923 strain and oxacillin-resistant S.aureus strain after 1 h of intervention (P=0.004 and P=0.026, respectively), which lasted to 5 h for ATCC25923 strain (P=0.012) and 3 h for oxacillin-resistant strain (P=0.001).Conclusion This study shows that rAtlM has a certain antibacterial effects on S.aureus ATCC25923 strain and oxacillin-resistant S.aureus strain, suggesting a possibility of serving as antimicrobial agent.

In the criminal cases of driving under the influence (DUI), DNA evidence can be collected from the deployed airbag of the motor vehicle and submitted to the crime lab for touch DNA analysis. The evidence can be acquired when the skin cells are observed on the surface of the airbag in a traffic accident. However, the low quantity or quality of the evidence collected from a crime scene prevents further identification analysis in many cases. In the current study, we reported a case of identifying touch DNA extraction from the shed skin cells from the deployed airbag of a motor vehicle. We managed to collect DNA evidence from the shed skin cells in an airbag using a proper approach of collection and extraction. The 5.87 ng of extracted DNA was sufficient for genotyping and forensic identification, which helped to identify the driver of the car in collision with a pier in the street. In DUI cases and other traffic accidents, therefore, the amount of touch DNA extracted from the deployed airbag can be sufficient for DNA marker genotyping and further analysis.
Accidents, Traffic ; Air Bags ; Alcoholic Intoxication ; Crime ; DNA/analysis* ; Genotype ; Humans ; Motor Vehicles ; Skin/cytology* ; Touch

OBJECTIVEThe present study is to observe in vitro the proliferation ability of the muscle cells from permanent myopathy (PM) patients of nomokalaemic periodic paralysis (normKPP), which is caused by mutations of Met1592Val in the skeletal muscle voltage gated sodium channel (SCN4A) gene on chromosome 17q23.1. We also evaluate the possible effect of the foreign basic fibroblast growth factor (bFGF) in preventing and curing PM.

METHODSThe gastrocnemius muscle cells were taken from two male patients with PM of the same Chinese family with Met1592Val mutation of SCN4A, determined by gene screening. Four male patients suffering from the skeletal injury without PM were taken as control. All preparations were protogenerationally cultured in vitro. Proliferation of the cultured preparations was measured by MTT. Activities of the lactic dehydrogenase (LDH), creatine kinase (CK), and protein content in these cells were also detected. The effects of bFGF with different doses (10 ng/mL, 20 ng/mL, 40 ng/mL, 80 ng/mL, 120 ng/mL and 160 ng/mL) on the above mentioned parameters were also evaluated.

RESULTSCells from both PM and control subjects were successfully cultured in vitro. The cultivation of the muscle cells from PM patients in vitro was not yet seen. Results indicated the obvious stimulation of bFGF on cell proliferation, activities of LDH and CK, protein synthesis, in a dose dependent manner. The optimal dose of bFGF was 120 ng/mL (P<0.05), beyond which greater dose caused a less effect. The effect of bFGF on 160 ng /mL was stronger than that on 80 ng/mL, but there was no significant difference (P>0.05).

CONCLUSIONMyoblastic cells from patients with PM had a weaker ability of developing into the myotubules, thus they were unable to perform effective regeneration, which resulted in a progressive necrosis. The exogenous bFGF could promote the division and proliferation of the muscle cells in vitro. These results shield a light on bFGFos potential role in preventing and treating PM.

Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Creatine Kinase ; metabolism ; Dose-Response Relationship, Drug ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Male ; Methionine ; genetics ; Middle Aged ; Muscle Development ; genetics ; physiology ; Muscular Diseases ; genetics ; pathology ; Mutation ; genetics ; Myoblasts ; drug effects ; NAV1.4 Voltage-Gated Sodium Channel ; Sodium Channels ; genetics ; Valine ; genetics

Abstract: Objective　To investigate the drug demand and related influencing factors of AIDS non-occupational post-exposure prophylaxis (nPEP) among men who have sex with men (MSM) in Wuhan, and to provide a scientific basis for the development of subsequent intervention policies for MSM. Methods　With the assistance of social organizations in Wuhan, MSM was recruited by the snowball method to carry out an online questionnaire survey to collect information on demographics, AIDS-related knowledge, high-risk behaviors, and the need for nPEP medication. The χ2 test and unconditional Logistic regression were used to analyze the related factors of the demand for nPEP medication. Results　A total of 308 valid subjects were included in this study, with predominantly 18-29 years old (78.57%, 242/308). The self-reported sexual orientation was mainly homosexuality (82.47%, 254/308), and the awareness rate of AIDS knowledge was high (89.29%, 275/308). Among the survey respondents, 35.06% (108/308) did not know the situation of HIV infection among MSM population in Wuhan; 55.19% (170/308) had two or more same-sex sexual partners in the last six months; 90.91% (280/308) had heard of nPEP before participating in this survey. After passing nPEP and informing the protective effect of nPEP, 59.42% (183/308) of them needed nPEP. After HIV exposure, 73.38% (226/308) were willing to spend money to buy nPEP drugs, and 88.64% (273/308) were willing to take nPEP drugs because of the known side effects. Logistic regression analysis showed that in the last six months, the needs for taking nPEP medication in those who had 2 or more same-sex sexual partners (OR=2.121, 95%CI: 1.329-3.386) and who had received peer education (OR=1.740, 95%CI: 1.088-2.781) were higher than those of those who had a same-sex sexual partner in the last six months and who had not received peer education. Conclusions　The MSM population in Wuhan has a great demand for nPEP drugs, and peer education is an important way to carry out nPEP publicity and promotion. At the same time, we should continue to strengthen warning publicity and education and behavioral intervention to reduce MSM risky sexual behaviors and reduce new HIV infections.

Dental hygienist is not only an important system of stomatology education, but also an important process of the clinical diagnosis and treatment in Japan. This paper discusses the development and current situation of dental hygienist system in Japan, and mainly introduces the admission requirements, teaching courses, employment status and clinical work, which maybe provide some important references for stomatology education and dental clinic for us in such aspects as specialty setting, talents cultivation and clinical team cultivation.

OBJECTIVETo investigate the relationships between rs3865418 polymorphism of neural precursor cell expressed developmentally downregulated 4-like gene and obesity in Kazakh general population.

METHODSBased on a cross-sectional epidemiological study in a Kazakh general population, a case-control study was conducted. The rs3865418 polymorphism in a Kazakh general population (856 subjects, including 364 males and 492 females; 478 in obesity group and 378 in normal control group) was genotyped by TaqMan polymerase chain reaction, and the relationship between rs3865418 polymorphism and obesity was analyzed.

RESULTSThe rs3865418 polymorphism was successfully genotyped in 851 Kazakh subjects. The distribution of the genotypes and alleles of rs3865418 polymorphism did not differ significantly between the obesity group and normal control group in terms of general populations, males, and females (all P > 0.05). The waist circumference showed a tendency of C/C > C/T > T/T in males and C/C < C/T < T/T in females, but without statistical significance (P > 0.05).

CONCLUSIONSThe rs3865418 polymorphism of neural precursor cell expressed developmentally downregulated 4-like gene may not be associated with obesity in Kazakh general population. In other words, it is not a predisposing factor for obesity in Kazakh.

Adult ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; China ; Cross-Sectional Studies ; Endosomal Sorting Complexes Required for Transport ; genetics ; Ethnic Groups ; genetics ; Female ; Genetic Predisposition to Disease ; Humans ; Male ; Middle Aged ; Nedd4 Ubiquitin Protein Ligases ; Obesity ; genetics ; Polymorphism, Genetic ; Ubiquitin-Protein Ligases ; genetics

Objective To explore the biological exposure limit of liver dysfunction induced by arsenic-coal burning, and screen sensitive biornarkers for its' liver dysfunction monitoring. Methods One hundred and eighteen subjects from the exposed area and 50 control from non-pollution area were studied. Their urinary and hair contents of arsenic were tested as exposure biomarkers by Ag-DDC assay. Total bile acid(TBA, detected by enzymatic cycling method), glutathione S-transferase (GSTs, detected by chemical colorimetry) and γ-glutamyl transpeptidase (γ-GT, detected by colorimetry of diazotization reagent) were used as biomarkers indicating liver cell damage. were used as liver fibrosis biomarkers. The benchmark dose (BMD) and the lower confidence limit of benchmark dose(BMDL) of urinary and hair arsenic were calculated. Sensitivity of each biomarker was estimated according to the BMD and BMDL value. Results The geometric mean of urinary and hair arsenic(98.50 mg/kg Cr, 7.42 mg/kg) μg/L) in the exposed group were significantly higher than urinary and hair arsenic (22.98 mg/kg Cr, 1.28 mg/kg) and each biomarker in the control group(4.63 μmol/L, 13.76 U/L,36.45 U/L,54.62 μg/L,74.45 μg/L,54.81 μg/L, P<0.01). Significant dose-effect relationship existed between urinary and hair arsenic contents and each biomarker. BMD and BMDL value of urinary arsenic was 49.53-101.96 mg/kg Cr and 39.02-70.15 mg/kg Cr, respectively. Those of hair arsenic were 3.04-5.02 mg/kg and 2.36-3.25 mg/kg, respectively. According to BMD and BMDL value of urinary and hair arsenic, the sensitivity of biomarkers decreased in the order of GSTs, TBA and Conclusions According to the lowest BMD and BMDL of urinary and hair arsenic, averaged reference value of urinary and hair arsenic in the local normal population, we suggest urinary 35.0 mg/kg Cr and hair 2.5 mg/kg as their biological exposure limits for those with liver dysfunction induced by arsenic-coal burning. GSTs, TBA, γ-GT and HA, Ⅳ. C, PC-Ⅲ can respectively reflect liver cell damage and liver fibrosis caused by arsenic-coal burning in different degrees, among which, GSTs and HA are the most sensitive biomarkers respectively for liver cell damage and liver fibrosis.