Benzene ; toxicity ; Gene Expression Profiling ; Oligonucleotide Array Sequence Analysis

Objective To explore the molecular types of methicillin-resistant Staphylococcus aureus (MRSA) strains present in major hospitals in Qingdao area, using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) methods, trying to find out the epidemiological characteristics of these MRSA isolates. Correlation of the PFGE types with microbiological phenotypes and clinical data was also studied. Methods 360 isolates of MRSA were procured during 2003 to 2007 from major hospitals in Qingdao. PFGE technology was applied to comparatively analyze the chromosomal DNA digested with endonuclease Sma Ⅰ . Comparison of DNA fragments patterns from each MRSA strain and cluster analysis were performed with the Bionumericus version ' 2.0' software. A dendogram was generated using PFGE macrorestriction fragments on gel images. Data was used to predict the possibility of each PFGE type via SPSS software version 11.0, using the variables as predictors including groups on patient's age, gender, source and the site where MRSA was isolated. Antibiotic sensitivity patterns of these MRSA isolates were determined by K-B tests, and a correlation between these patterns and PFGE types was investigated. Housekeeping genes were amplified with PCR and sequenced in representative strains of variant PFGE types to identify their allelic profile. Results 5 types of PFGE patterns (M0-M4) were identified with MI being the predominant and M2 next to it which was significantly correlated to the isolates from wounds. M3 type strains were mainly isolated from ICU wards and there were a few cases complied with M4 type with no correlated variant factors found in this study. A unique pattern of MRSA isolates with its M0 distinct from other types had not been reported. No significant association was found between PFGE individual types,gender or age groups. M1 and M2 types were the major proportional PFGE patterns among different hospitals. No vancomycin-resistant isolates were detected among 360 MRSA strains. No significant association was found between individual antibiotic resistance and specific PFGE types. Data from MLST analysis showed that the aUelic profiles of M1 and M3 type strain had the same ST239 linage which was commonly present in China. For M2 and M4 representative strains, the allelic profiles were ST5 and ST240, respectively. ST45 and ST398 were corresponding to two PFGE patterns clustered as M0 type. Conclusion Nosocomial infection due to MRSA was evenly distributed among different age groups and no gender bias was observed. The PFGE types of MRSA strains isolated in major hospitals in Qingdao were highly correlated with the sources of isolates and ST239 isolate seemed the prevalent and widespread one. Strategies should be designed to further monitor and prevent or minimize the spread of ST5 MRSA isolates and the like, in Qingdao area.

OBJECTIVETo identify triacylglycerols in coix oil.

METHODHigh performance liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry was used for identification. The experiment was operated under the conditions: spray voltage at 3 000 V, capillary temperature at 250 degrees C, APCI vaporizer temperature at 400 degrees C, and corona current of 4 microA. Sheath gas pressure (high purity liquid nitrogen) was 35 kPa. Mass spectra were obtained over the m/e range of 300 to 900 amu, scan duration of 1s and Q1 peak width at 0.7. The stationary phase was Zorbax Extend C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase: dichloromethane-acetonitrile (35:65), flow rate: 1 mL x min(-1); column temperature: 25 degrees C.

RESULT12 triacylglycerols were identified by HPLC-MS method.

CONCLUSIONThe result can be used to identify the components in a fingerprint chromatogram of coix oil and its related injection product.

Chromatography, High Pressure Liquid ; methods ; Coix ; chemistry ; Mass Spectrometry ; methods ; Plant Oils ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Seeds ; chemistry ; Triglycerides ; analysis ; chemistry ; isolation & purification

BACKGROUNDSolar ultraviolet (UV) irradiation induces the production of matrix metalloproteinases (MMPs) by activating cellular signalling transduction pathways. MMPs are responsible for the degradation and/or inhibition of synthesis of collagenous extracellular matrix in connective tissues. We mimicked the action of environmental ultraviolet on skin and investigated the effects of UVB-irradiated human keratinocytes HaCaT and IL-1alpha on mitogen activated protein (MAP) kinase activation, c-Jun and c-Fos (AP-1 is composed of Jun and Fos proteins) mRNA expression and MMP-1 production in UVA-irradiated dermal fibroblasts.

METHODSFollowing UVA irradiation, the culture medium of fibroblasts was replaced by culture medium from UVB-irradiated HaCaT, or replaced by the complete culture medium with interleukin (IL)-1alpha. MAP kinase activity expression in fibroblasts was detected by Western blot. c-Jun and c-Fos mRNA expressions were determined by reverse transcriptional polymerase chain reaction (RT-PCR); MMP-1 production in culture medium was detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSCulture medium from UVB-irradiated keratinocytes increased MAP kinase activity and c-Jun mRNA expression in UVA-irradiated fibroblasts. IL-1alpha increased MAP kinase activity and c-Jun mRNA expression, IL-1alpha also increased c-Fos mRNA expression. Both culture media from UVB-irradiated human keratinocytes and externally applied IL-1alpha increased MMP-1 production in UVA-irradiated fibroblasts.

CONCLUSIONSUVB-irradiated keratinocytes and IL-1alpha indirectly promote MMP-1 production in UVA-irradiated fibroblasts by increasing MAP kinase/AP-1 activity. IL-1 may play an important role in the paracrine activation and dermal collagen excessive degradation leading to skin photoaging.

Cell Line ; Enzyme Activation ; Fibroblasts ; enzymology ; radiation effects ; Humans ; Interleukin-1 ; pharmacology ; Keratinocytes ; physiology ; radiation effects ; Matrix Metalloproteinase 1 ; biosynthesis ; Mitogen-Activated Protein Kinases ; metabolism ; Proto-Oncogene Proteins c-fos ; genetics ; Proto-Oncogene Proteins c-jun ; genetics ; RNA, Messenger ; analysis ; Skin ; radiation effects ; Skin Aging ; Transcription Factor AP-1 ; metabolism ; Ultraviolet Rays

Adolescent ; Child ; Child, Preschool ; Chromosomes, Human, Pair 6 ; Female ; Humans ; Infant ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics

To study still further the activity of CNTF mutant designed by computer molecular modling,the methods of dissociated cultures of chick dorsal root ganglion、TF-1 prolification and the normal mice'weight loss tests weve used.The results indicated that the mutant protein promoted the survival of dorsal root ganglion、induced TF-1 prolification and made the normal mice lose weight,decrease appetite and reduce fat index.The weight loss effect was dependant with its administration dosage,ED50 was 150.986?g/kg/d.To TF-1,the specific activity reached 2.0?106U/mg against international reference reagent.In a word,CNTF mutant had excel bioactivity.So it provided clues for its development and application.

OBJECTIVETo establish a new method of multiplex semi-nested polymerase chain reaction (PCR) to detect pathogens in cerebrospinal fluid (CSF).

METHODSAccording to the analysis of the conservative and variable regions in bacterial 16S rRNA genes, we designed universal primers for all bacteria and specific primers for most gram-positive and gram-negative bacteria. All primers were added into the same reaction systems successively of a two-step PCR assay to amplify the different bacterial DNA in CSF, and the results were compared with common culture method with sensitivity and the specificity both detected at the same time.

RESULTSBoth gram-positive and gram-negative bacteria amplified DNA fragment about 1,032 bp after first-step amplification with universal primers. In the second step, specific fragments of 336 bp and 127 bp were amplified in gram-positive and gram-negative bacteria respectively besides fragments of 1,032 bp; The detection limit for E. coli was 8 cfu/ml. The comparison of 62 CSF samples detected by both multiplex semi-PCR and conventional culture method revealed sensitivity, specificity, positive and negative values of 93.8%, 95.7%, 88.2%, and 97.8% respectively for PCR.

CONCLUSIONThe result suggested that the multiplex semi-nested PCR we established was sensitive, specific and rapid method for clinical laboratory to detect pathogens in CSF.

Cerebrospinal Fluid ; microbiology ; DNA Primers ; genetics ; DNA Probes ; genetics ; DNA, Bacterial ; cerebrospinal fluid ; isolation & purification ; Escherichia coli ; genetics ; isolation & purification ; Gram-Negative Bacteria ; genetics ; isolation & purification ; Gram-Positive Bacteria ; genetics ; isolation & purification ; Humans ; Polymerase Chain Reaction ; methods ; RNA, Bacterial ; genetics ; RNA, Ribosomal, 16S ; genetics ; Sensitivity and Specificity ; Staphylococcus aureus ; genetics ; isolation & purification

OBJECTIVETo compare the clinical efficacy differences between fire needling technique of filiform needle at high stress points and regular acupuncture on chrondromalacia patellae so as to provide the better therapy for the treatment of this disease.

METHODSSixty cases of chrondromalacia patellae were randomized into a fire needling group (28 cases) and a routine acupuncture group (32 cases). In the fire needling group, 5 to 6 high stress points were localized according to the symptoms, palpation and imaging condition and were stimulated with fire needling technique of filiform needle. The treatment was given once every two days, 5 treatments made one session. In the routine acupuncture group, the regular acupuncture was applied at Dubi (ST 35), Xiguan (LR 7), Yanglingquan (GB 34), Yinlingquan (SP 9), Zusanli (ST 36), etc. The treatment was given once every day, 5 treatments made one session. Lysholm score, VSA score, patella title angle (PTA) and lateral patella angle (LPA) of the affected knees before and after treatment, as well as the clinical efficacy after treatment were observed in the two groups.

RESULTSAfter treatment, Lysholm score, VSA score, PTA and LPA were all improved apparently in the two groups (all P < 0.01). After the treatments, the improvements in Lysholm score, VSA score, PTA and LPA in the fire needling group were more obvious than those in the routine acupuncture group (all P < 0.05). The effective rate was 92.9% (26/28) in the fire needling group, better than 87.5% (28/32) in the routine acupuncture group (P < 0.01).

CONCLUSIONThe fire needling technique of filiform needle at the high stress points relieves the clinical symptoms of chrondromalacia patellae and recovers the biodynamical structure of patellae.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Adult ; Chondromalacia Patellae ; therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo determine the main genotype of hepatitis B virus (HBV) in Xinjiang.

METHODS200 HBsAg positive serum specimens were detected from more than 2000 serum of Xinjiang inhabitants, and HBV S gene was detected by using nPCR amplifying, and compared with the standard S region HBV nucleotide sequences of genotypes A-H retrieved from GenBank, then analyzed and drawn the polygenetic tree by MEGA3 software.

RESULTGene in 127 (63.5%) serum specimens was detected from 200 samples. Among 127 serum specimens, 10 (7.8%) was genotype B, 58 (45.7%) was genotype C, and 59 (46.5%) was genotype D.

CONCLUSIONGenotype B, C and D have been found in Xinjiang.

Adolescent ; Adult ; Aged ; Child ; China ; Female ; Genotype ; Hepatitis B ; ethnology ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Rural Health ; Young Adult

OBJECTIVETo establish a cell line for stable expression of human beta-defensin 3 (hBD3).

METHODSFull length cDNA of hBD3 was isolated from previously constructed pGEM-hBD3 and then inserted into pcDNA3. The recombinant vector identified carrying hBD3 with right direction was introduced into COS-7 cells by Lipofectamine. Cell clones survived in G418-rich medium and with stable expression of hBD3 in both mRNA and protein levels were identified by RT-PCR and Western blot respectively. Genomic integration of the hBD3 gene with the COS-7 cells was confirmed by Southern dot blot and primary analysis. The antimicrobial activity of the secreted hBD3 was also evaluated.

RESULTSCOS-7 cells transfected with pcDNA3-hBD3 expressed hBD3 stably in mRNA and protein level. Southern dot blot analysis showed successful integration of the hBD3 gene into the genome of COS-7 cell and the hBD-3 protein secreted into the culture medium showed antimicrobial activity.

CONCLUSIONWe successfully established a hBD3-expressing cell line.

Animals ; COS Cells ; Cercopithecus aethiops ; Escherichia coli ; drug effects ; Genetic Vectors ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Staphylococcus aureus ; drug effects ; Transfection ; beta-Defensins ; biosynthesis ; genetics ; pharmacology