Objectives To compare the difference of ERG11 partial promoter sequence(-440 ～ -1)between yeast-form and hypha-form of Candida albicans, and the relation between the mutation of ERG11 promoter and the susceptibility of C. albicans to fluconazole. Methods Yeast-form and hypha-form of C. albicans genome DNA were extracted from 4 substrains C. albicans, which were isolated from a single HIV-sero-positive patient. Partial ERG11 promoters of yeast-form and hypha-form of these 4 substrains were amplified by polymerase chain reaction (PCR) and the PCR products were sequenced. Results There was no sequence difference between the yeast-form and the hypha-form of C. albicans in this region. Heterogeneities at position -365, -353, -328, -310, -308, -299, -295, -293, -292, 290, -289 and a single base deletion (-284 delT) were found on one allele of ERG11 promoter region of fluconazole-susceptible substrains. No heterogeneities were observed in dose-dependent fluconazole-susceptible and fluconazole-resistant substrains. Conclusions There is no difference of ERG11 partial promoter sequence between the yeast-form and the hypha-form C. albicans. The mutation of ERG11 promoter may be associated with the resistance of C. albicans to fluconazole.

To summarize the application of proteomics to modernization research of Chinese materia medica (CMM) and offer the reference for modernization of CMM and new medicine preparation exploitation. Based on the references and our project group studies in proteomic of drug biosynthesis pathway and metabolization;the main content,technology strategies and important application of the proteomics were introduced significantly;and the further development of protemics was analyzed. The proteomics will become one of absolutely necessary tackles in the modernization research of CMM.

AIM: To access the difference of the efficacy between terbinafine and itraconazole in the treatment of dermatophyte onychomycosis. METHODS: The Medline, Science Direct On Site (SDOS), and Springer database were searched in detail on the data of the mycological cure rates of the two antifungal agents for treatment of dermaphyte onychomycosis occourring in patients aged from 18 a to 60 a with the published double blind randomized clinical trials and then pooled. The odds ratio (OR) and its 95 % confidence interval (CI) were calculated. RESULTS: Six treatises of double blind randomized clinical trials were selected for this analysis according to the screening criteria. The mycological cure rate of continuous terbinafine 250 mg per day was higher than that of either therapeutic effect of itraconazole pulse 400 mg per day (OR = 5.01, 95 % CI (3.42 - 7.33)) or continuous itraconazole 200 mg per day (OR = 2.58, 95 % CI (1.91 - 3.49)) . CONCLUSION: Terbinafine is more effective than itraconazole in the treatment of dermatophyte onychomycosis.

Objective To compare 2 approaches with different culture media which induce hyphal form of Candida albicans.Methods Induction of hyphal form was conducted for 16 C.albicans strains with either RPMI 1640 medium or DMEM medium,at 37 ℃ for 24 h,respectively.The hyphal and yeast forms were counted separately and the ratio of hyphal form to total cells was calculated.Results The ratio of hyphal form to total cells was higher in RPMI 1640 medium than that in DMEM medium at the same incubation time for the majority of strains.The ratio was above 99% for all strains after 7-day incubation with 12 times of passages in RPMI 1640 medium at 37 ℃.Moreover the ratio of hyphal form was significantly higher for fluconazole-susceptible strains than that for fluconazole dose-dependent susceptible and resistant strains in incubation with DMEM medium at 37 ℃.Conclusion Incubation with RPMI 1640 medium at 37 ℃ for 7 days seems a favorable condition to induce hyphal form of C.albicans.

to common antifungal drugs.

Objective To construct a mutant strain of Aspergillus fumigatus with the mutant pbs2 gene, and to investigate the effect of the mutant gene on the morphology of A. fumigatus, the variation of os-motic pressure and the sensitivity of oxidative stress. Methods S. cerevisiae pbs2 gene homologs was identi-fied in A. fumigatus genome database. A PCR fragment, composed of the 5' flanking sequence(approximate-ly 1 kb) ofpbs20RF and its 3' flanking sequence (approximately 1 kb), was subcloned into a vector pDHt/SK with Xho Ⅰ/Xba Ⅰ enzyme to produce a recombinant plasmid PA. The selected marker pryG amplified from pLAX223 was cloned into PA with BamH Ⅰ/Pst Ⅰ enzyme to produce plasmid PB. PB was transformed into Agrobacterium tumefac/ens and named plasmid At pbs2. Then At pbs2 was transformed into A. fumigatus strain AF293.1 (pyrG) via ATMT to produce the transformants △pbs2. To observe the growth rate and the phenotype on the MM containing different oxidative stress and osmotic pressure between the △pbs2 and the wild type. Results There were no visible difference between △pbs2 and AF293 on the radial growth rate and morphology. △pbs2 mutant was not only more sensitive to osmotic pressure produced by sodium chloride and glycerol but also sensitive to oxidative stress produced by hydrogen peroxide than the wild type. Conclu-sion The pbs2 gene plays a role in osmotic pressure and oxidative stress signal tmnsductions in A. fumiga-tus but only involved in oxidative stress induced by hydrogen peroxide specifically. However, pbs2 gene has a minor effect on growth and morphology in A. fumigatus.

Objective:To construct the sho1 gene mutant of Aspergillus fumigatus,and to investigate the function of sho1 in adaptation to environmental changes.Methods:Saccharomyces cerivisiae sho1 gene homolog was identified by tblastn searches in A.fumigatus genome database.A PCR fragment,composed of the 5' flanking sequence(approximately 1 kb) of sho1,sho1 ORF,and its 3' flanking sequence(approximately 1 kb),was subcloned into a vector pDHt/SK2 with ApaⅠ/Xba Ⅰ enzyme to produce a recombinant plasmid PA.The selected marker pyrG cut from pLAX223 with Nco Ⅰ/Pst Ⅰ was cloned into PA,and PB was produced.The plasmid PB was transformed into competent Agrobacterium tumefaciens EHA105 by using the freeze/thaw method.The strain of A.tumefaciens was designated as At sho1.Then sho1 was produced via ATMT.The radial growth was observed and compared between sho1 and wild type(wt) on MM,CM and BHI medium,and the sensitivity of sho1 and wt to amphotericin B,itraconazole,voriconazole,and caspofungin investigated.Results:Radial growth had visible difference between sho1 and wt on MM,CM and BHI medium.sho1 mutant was sensitive to high osmotic pressure but not to low osmotic pressure.The susceptibility to antifungal drugs was similar between the sho1 and wt.Conclusion:Slow growth of the sho1 mutant has nothing to do with the different medium.The sho1 gene plays a role in response to osmotic pressure in A.fumigatus,but sho1 has a minor role in the susceptibility to the antifungal drugs despite the slow growth.

Objective:To investigate the efficiency of Agrobacterium tumefaciensmediated transfor-mation of Aspergillus fumigatus by using pyrG as a recessive selectable marker.Methods: FAP1 and SHO1 genes target sequences,composed of a selectable marker pyrG and the flanking sequences of the FAP1 and the SHO1 genes,were cloned into a binary plasmid pDHt/sk,respectively.The produced plasmids were transformed into A.tumefaciens.The A.tumefaciens and uracil auxotroph A.fumigatus were cocultured in induction medium without uricil and uridine at 24 ℃ for 48 h.To inhibit growth of A.tumefaciens and to select transformants,the cultures were transferred to 37 ℃ and incubated for another 48 h.Results: In this study,A.tumefaciens-medidated transformation of A.fumigatus produced high homologous recombination rates,which was 44%(7 of 16) for FAP1 and 35%(7 of 20) for SHO1.Conclusion: Our study showed that A.tumefaciens-medidated transformation by using pyrG as a recessive selectable marker is an efficient tool for target gene deletion of A.fumigatus.

Objective To identify clinicopathologic factors which predict survival following hepatectomy in HBV-related cirrhotic patients with early hepatocellular carcinoma (HCC).Methods A database was used to identify patients with histologically confirmed early HCC (≤5 cm,no nodal involvement,metastases,or major vascular invasion) who underwent surgical resection (excluding ablation or transplantation).Among 20 700 patients with HCC who were diagnosed at the Eastern Hepatobiliary Surgery Hospital from April 2005 to November 2010,537 (2.6％) patients with early HCC were studied retrospectively.Prognostic factors were evaluated using the Kaplan-Meier curves,Cox proportional hazards models and the receiver operating characteristic (ROC) curves.Results The study included 537 patients.The median tumor size was 2.9 cm,and 33％ of patients had tumors ≤2 cm.Most HCC lesions were solitary (63％) and had no evidence of vascular invasion (64％).Following surgery,the overall median and 5-year survival were 45 months and 33％ respectively.After adjusting for demographic factors and histological grade,tumor size ＞2 cm (hazard ratio [HR]:1.56),multifocal tumors (HR:1.34),and vascular invasion (HR:2.03) remained independent predictors of poor survival (all P ＜ 0.05).Based on these findings,a prognostic scoring system was developed that allotted 1 point each for these factors.Patients with early HCC could be stratified into 4 distinct prognostic groups (median and 5-year survival,respectively):0 points (97 months,96％),1 point (85 months,76％),2 points (76 months,54％),3 points (56 months,39％) (P ＜0.01).Conclusions The present study emphasized the importance of pathologic staging even in patients with small HCC.Anatomical resection of HCC should be the preferred surgical procedure in cirrhotic patients.

Basic surgical skills tralning which belongs to the basic course of surgery, is clini-cal basis for the medical students. The teaching practice of basic surgical skills tralning was explored by the Affiliated Hospital of Inner Mongolia Medical University, the theoretical teaching was combined with skills tralning practice, three kinds of means which contalned the model, animal tissues and or-gans in vitro and animal experiment were used to carry out systemic and standardized tralning. Finally, the basic surgical skills of postgraduate in surgery were improved, and the expected alm of teaching was achieved.