ABSTRACT
In order to provide a rational research basis for clinical detection and genetic engineering vaccine, plasmid pET-28a (+) encoding both Porin gene PIA and PIB of Neisseria gonorrhoeae was constructed and a fusion protein in E. coli DE3 expressed. The fragments of PIA and PIB gene of Neisseria gonorrhoeae were amplified and cloned into prokaryotic expression plasmid pET-28a (+) with double restriction endonuclease cut to construct recombinant pET-PIB-PIA. The recombinant was verified with restriction endonuclease and sequenced and transformed into E. coli DE3 to express the fusion protein PIB-PIA after induced with IPTG. The results showed PIA-PIB fusion DNA fragment was proved correct through sequencing. A 67 kD (1 kD= 0.992 1 ku) fusion protein had been detected by SDS-PAGE. It was concluded that the fusion protein was successively expressed.
Subject(s)
Humans , Cloning, Molecular , Escherichia coli , Genetics , Metabolism , Neisseria gonorrhoeae , Genetics , Plasmids , Genetics , Porins , Genetics , Prokaryotic Cells , Metabolism , Recombinant Fusion Proteins , Genetics , Transfection , Vaccines, SyntheticABSTRACT
Objective To investigate the expression of intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1(VCAM-1) on the surface of human vascular endothelial cells (HUVECs) stimulated by Candida albicans.Methods HUVECs were isolated from the umbilical vein of placenta of healthy parturients and cultured.The primary culture of HUVECs was stimulated with Candida albicans at different intervals.Expression of ICAM-1 and VCAM-1 mRNA were measured by reverse transcription-PCR,and the levels of secretion of IL-6 and IL-8 by enzyme-linked immunosorbent assay(ELISA).Results ICAM-1 and VCAM-1 mRNA expression in HUVECs began to increase after stimulation with C.albicans in 4 h and reached peak level in 8 h.Secretion of IL-6 and IL-8 by HUVECs began to increase after stimulation by C.albicans in 4 h and reached peak level in 24 h.Conclusion C.albicans could induce expression of ICAM-1 and VCAM-1 mRNA in HUVECs,and enhance secretion of IL-6 and IL-8 by HUVECs.
ABSTRACT
Fungus infection of the patients with chronic prostatitis was studied and the applied value of modified Sabourand's liquid medium in fungus culture explored. The causative fungus in expressed prospapic secrepion (EPS) of chronic prostatitis patients were examined by improved Sabouraud's liquid medium and Sabouraud's glucose agar medium. The fungus species were identified. The results showed that in 1138 cases positive for fungus，the specimens of 95 cases (8.35 %) were cultured in the improved Sabouraud's liquid medium and those of 30 cases (2.64 %) in Sabouraud's glucose agar medium. In the 95 cases，63 strains were Candida albicans (63.3 %)，15 Candida tropicalis (15.8 %)，and 17 other Candida (17.9 %). It was concluded that: (1) The relationship between chronic prostatitis and fungus infection can not be ignored. The attention should be paid to the possibility of fungus infection for those with chronic postatitis who were treated with long-time antibiotics but the curative effects were not satisfactory;(2) In Candidal prostatitis，Candida albicans was a common causative fungus;(3) Application of the improved Sabouraud's liquid medium could increase the positive rate of Candida.
ABSTRACT
Objective: To explore the infection of mycoplasma in the venereal diseases and information about their drug susceptibility. Methods: Materials from the 12 528 patients were cultivated and drug susceptibility made testing was made using the mycoplasma IST Kit. Results: The results showed that 9433 of 12 528 (75.3%)were infected by mycoplasma, Uu infective rate 58.3 %(7 305 cases)was found significantly higher than Mh infective rate 3.0%(379 cases)and mixed infection of both Uu and Mh was 14.0%(1 749 cases,)P＜0.01. The no susceptible conditions were in turn that ofloxacine(50.5%), Erythromycine(41.9%), tetracycline(14.6%), doxycycline(4.7%), josamycine(2.9%), and pristinamycine(2.0%) respectively. Conclusions:The results suggested that non-gonococcal genitourinary tract infection was firstly ureaplasma urealyticum, concentracted infection was at age 21-40 years. no susceptible stat of mycoplasma infection to drug was serious, so that susceptibility testing of mycoplasma had a important significance to clinical drug treatment.