Objective:To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on ceil proliferation and apoptosis after transferred into SW1990 cell line.Methods:K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR.Clones with inverted insertion were selected and co-transferred into E.coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination.Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus.Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified;the virus titer was determined.Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining.SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay.Results:A 282 bp target gene fragment was acquired by PCR;the titer of recombinant adenovirus was 7.6?10~8 pfu/ml before purification by CsCl_2 gradient centrifugation and 5.0?10~(10)pfu/ml after CsCl_2 gradient centrifugation.When the recombinant adenovirus was at 100 MOI,the infection efficiency of SW1990 cells nearly reached 100%.The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation(P

OBJECTIVETo investigate the correlations of 24 biochemical markers in the seminal plasma with routine semen parameters.

METHODSAccording to the WHO5 standards, we analyzed the routine semen parameters of 66 subfertile men, including the semen volume, sperm concentration, total sperm count, sperm motility, and the percentage of progressively motile sperm (PR). Based on the calibration and quality control measures and using an automatic biochemistry analyzer or electrolyte analyzer, we detected 24 biochemical markers in the seminal plasma of the patients, including total protein (TP), albumin (Alb), globulin (Glb), uric acid (UA), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), γ-glutamyltransferase (GGT), lactate dehydrogenase (LDH), creatine kinase (CK), alpha hydroxybutyrate dehydrogenase (αHBDH), adenosine deaminase (ADA), glucose (Glu), triglyeride (TG), total cholesterol (TC), urea nitrogen (UN), creatinine (Cr), high-sensitive C-reactive protein (hsCRP), K+, Na+, Cl- , Ca, Mg, and phosphorus (P). Then we analyzed the correlations of the 24 biochemical markers with routine semen parameters.

RESULTSThe levels of the TP, Alb, and Glb proteins in the seminal plasma were positively correlated with sperm concentration, so was that of Alb with the total sperm count, and the AST and LDH activities with sperm concentration and total sperm count. The AKP activity in the seminal plasma was correlated negatively with the semen volume, but positively with sperm motility. The αHBDH activity exhibited a positive correlation with both sperm concentration and total sperm count, with a coefficient of correlation (r) above 0.7. The UN level was correlated negatively with the semen volume, so was the Cr level with the semen volume, sperm concentration, and total sperm count, and the Glu level with sperm concentration and total sperm count. The TG level was correlated positively with the semen volume, but negatively with sperm motility. The levels of seminal plasma ALT, GGT, ADA, UA, TC, CK, and hsCRP showed no correlation with the above-mentioned semen parameters. None of the seminal plasma K+, Na+, Ca, Mg, and P levels was found correlated with semen parameters except the Cl- level, which was negatively correlated with the semen volume.

CONCLUSIONMany biochemical markers in the seminal plasma are closely related to routine semen parameters, indicating that these biochemical components may play roles in spermatogenesis, sperm maturation, and physiological metabolism.

Biomarkers ; chemistry ; Humans ; Male ; Semen ; chemistry ; Semen Analysis ; Sperm Count ; Sperm Motility

Acupuncture Therapy ; Hemangioma, Cavernous ; surgery ; Humans ; Male ; Postoperative Complications ; therapy ; Spinal Cord ; blood supply ; Young Adult

Objective To investigate the difference between the blood pressure readings between sitting and supine position,and to study the factors that associated with the sitting-supine blood pressure difference in patient with diabetes.Methods We measured the sitting blood pressure first then followed by the supine pressure in 356 diabetic patients,using a standard mercury sphygmomanometer.Patient's body weight,height and blood glucose levels were also measured.Results SBP and DBP were significantly higher in the supine position than in sitting position in diabetic patients(by 3.5?7.6/1.5?4.9 mm Hg,P

Objective To detect the msr gene which confers resistance to erythromycin, and ana-lyze its distributing difference between the two biovars of Ureaplasma urealyticum. Methods Broth dilution method was used to determine the minimum inhibitory concentrations (MIC) to erythromycin among 72 U. urealyticum clinical isolates. The msrA, msrB, msrC and msrD genes detection and biotyping of U. urea-lyticum were conducted using PCR. Results The MICs of 72 U. urealyticum isolates to erythromycin ranged from ≤0. 125 μg/ml to ≥128 μg/ml. MIC_(50) was 32 μg/ml and MIC_(50) was ≥128 μg/ml. Biotyping showed that biovar Parvo had 51 strains (51/72, 70.83%) and biovar T960 had 21 (21/72, 29.17%) strains.The msrA, msrB, msrC and msrD genes were obtained in 1, 12, 0 and 24 strains, respectively, with five strains carrying the msrB and msrD genes, and one strain carrying the msrA, msrB and msrD genes. There was no resistance difference to erythromycin between the two biovars when the MIC≥8 μg/ml was considered resistance to eryt hromycin. But the msrB gene was predominantly detected in biovar T960. Conclusion U. urealyticum clinical isolates harbeur the msrA, msrB and msrD genes, and the predominantly detected msrB gene is of biovar T960.

Objective To study the relationship between erythromyein sensitivity and ermB gene in 143 Ureaplasma urealyticum (Uu) clinical isolates. Methods We detected the minimum inhabit concen-trations (MICs) of Uu to erythromycin by broth dilution method and MIC≥8 μg/ml was used as standard concentration of resistance to erythromycin. Polymerase chain reaction was used to detect the ermB gene and biotype Uu with primers based on multi-band antigen gene. Results The MICs, MIC50 MIC90 of Uu to erythromycin were ≤0. 125 μg/ml to ≥128 μg/ml, 16 μg/ml, and ≥128 μg/ml, respectively, with a high resistance rate of 64.38%. ermB gene, which was mainly detected in Uu with MIC≥8 μg/ml, was positively detected in 40 out of 143 Uu strains (27.97%). No significant differences of the resistance to erythromycin and positive rate of ermB gene were found between the two biovars in the study . Conclusion ermB gene may probably be one of the important genes conferring resistance to erythromycin in Uu. Further studies are needed to discover the difference of resistance and mechanism of erythromycin between the two bi-ovars.

Objective To study the resistance mechanism of Ureaplasma urealyticum (Uu) to erythromycin.Methods The susceptibility of 73 clinical isolates of Uu to erythromycin was evaluated by using broth dilution techniques. PCR and DNA sequencing were carried out to screen hot spot mutations at the variable region of 23S ribosomal RNA in erythromycin-resistant strains of Uu. Moreover, erythromycin resistance methylase genes (ermA, ermB, ermC) and efflux pump genes (mefA/E, msrA/B, mreA) were screened by using PCR with specific primers. Results There were 35 (47.95%) resistant Uu strains out of the 73 isolates, and the minimal inhibitory concentration varied from 8 to 32 mg/L among these resistant strains. The ermB gene was detected in 19 (54.29%) resistant strains, and msrA/B gene in 9 (25.71%) resistant strains. Two resistant strains harbored both ermB gene and msrA/B gene. No mutation at 23S ribosomal RNA or amplification of resistance-associated genes was noted in sensitive or reference strains of Uu. Conclusion The ermB and msrA/B genes may be responsible for the erythromycin resistance of Uu.

Objective To investigate the significance and distribution of lymphopenia and antilymphocyte antibody(ALA) in systemic lupus erythematosus (SLE) and to explore the role of ALA in lympho penia.Methods One hundred and ten in-patients who were admitted during February 2003 to February 2008 were retrospectively reviewed.Indirect immunofluorescence test was used to detect ALA.Results ① Lymphopenia (＜1.5×109/L) was observed in 68.2% patients.Lymphopenia was associated with skin rashes,serositis,renal involvement,NPSLE,leucopenia,ANA,ds-DNA antibody,ESR and IgG levels.The lymphocyte count and SLEDAI scores was more closely correlated than total white blood cell counts.②ALA was positive in 51/110 (46.4%) patients.ALA was associated with renal involvement,NPSLE,WBC count,C3 level decrease,ANA,dsDNA antibody and SLEDAI scores.Conclusion Lymphopenia is significantly associated with SLEDAI scores.ALA is possibly one cause of lymphcytopenia.In addition,ALA is also a parameter for disease activity,and is associated with organ involvement and outcomes.

Objective To study the ability of standard strain and clinical isolates of Ureaplasma spp. to form biofilms in vitro and to compare the antibiotic susceptibility of sessile cells and their planktonic counterparts. Methods A total of 21 Ureaplasma wealyticum(Uu) isolates recovered from female patients diagnosed with cervicitis and Uu serovar 3 and Uu serovar 8( Uu3, Uu8) were included. Scanning electron microscope and confocal scanning laser microscopy were used to identify biofilm formation. Conventional antibiotic susceptibility tests and biofilm susceptibility assays for tetracycline, erythromycin and ciprofloxacin were carried out. The paired rank sum test and was applied to analyze the statistical differences between the MIC and the minimal biofilm inhibitory concentration. The x2 test was applied to analyze the statistical differences of global resistance percentages between planktonic cells and sessile cells. Results Uu3, Uu8 and 21 Uu isolates all can form biofilms in vitro. Minimal inhibitory concentration of sessile cells compared with planktonic cells were obviously higher for tetracycline, erythromycin and ciprofloxacin (P <0.001). Global resistance percentages between planktonic cells and sessile cells were different for erythromycin (9.52% vs 61.90% , P < 0. 001), ciprofloxacin ( 80. 95% vs 100% , P = 0. 035 ) and tetracycline (4. 76% vs 14.29% , P =0.293). Conclusion Uu isolates and Uu1, Uu8 all can form biofilms in vitro, and biofilm formation can strengthen resistance of Uu to antibiotics, even multidrug resistance was observed.

OBJECTIVETo obtain Chlamydia trachomatis heat shock protein (cHSP) 10 gene from clinical secretion samples.

METHODScHSP10 gene was amplified from 20 cases of clinical secretion samples with positive gold-labelling by specific primers of cHSP10 and identified by sequence analysis.

RESULTScHSP10 full-length gene was amplified from 1 of 20 cases of clinical secretion samples with positive gold-labelling. cHSP10 gene encoding 102 amino acids contains 306 bp, which nuclotide at position 194 changes from T to A, leading to the change of corresponding amino acid.

CONCLUSIONScHSP10 gene may be cloned from clinical secretion samples with positive gold-labelling, which make it possible to further construct expression plasmid of recombinant cHSP10.

Antibodies, Bacterial ; blood ; Bacterial Proteins ; genetics ; Base Sequence ; Chaperonin 10 ; chemistry ; genetics ; immunology ; Chlamydia trachomatis ; chemistry ; immunology ; Cloning, Molecular ; Female ; Humans ; Infertility, Female ; etiology ; Male ; Molecular Sequence Data ; Polymerase Chain Reaction