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.

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 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 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 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.

A new strain of Streptomyces regensis was isolated from soil to produce a novel antibiotic AGPM possessing a strong antitumor activity In order to study on the metabolic path of the novel antibiotic AGPM, the protein patterns from the strain of Streptomyces regensis at different culture period were analyzed by using two eimensional polyacrylamide gel electrophoresis Comparing with sample from growth phase, seventeen new protein spots were found in that from antibiotic production phase The results demonstrated that the special proteins might be related with the antibiotic AGPM biosynthesis from Streptomyces regensis

In recent years,liver transplantation donor shortage as one of world medical problems is paid more attention by domestic and overseas scholars.In view of that,heterotopic auxiliary liver transplantation emerges which transplants the whole or some parts of a donor liver outside the original liver position on the condition that some parts or the whole of the original liver were retained.As for the liver transplantation with poor conditions on portal vein,reconstruction of portal vein has become an aporia.Based on this,some scholars put forward the theory——arterialization of portal vein(PVA),namely a method to increase arterial blood supply or replace portal vein blood perfusion of liver by establishing some pathes among artery and portal vein or its branches.The research background and current situation of heterotopic auxiliary liver transplantation with portal vein arterialization,the transplanting position of donor liver,vessel reconstruction,dynamics mechanism after reconstruction and liver regene-ration are summarized in this review.

Objective To investigate the appropriate cutoff point of CA19-9 in prognosis and to determine other potential prognostic factors which may affect survival of patients with hilar cholangiocarcinoma (HC) after radical surgery.Methods 168 patients who had undergone radical surgery for hilar cholangiocarcinoma with R0 and R1 resection were selected for the study.Categorized versions were used in univariate model to determine the appropriate cutoff point of CA19-9.CA19-9 and other clinicopathologic factors were analyzed for their influence on survival using multivariate methods.Results The strongest univariate predictor among the categorized preoperative CA19-9 measures was CA19-9 of less than 150 IU/L (P ＜0.001).On univariate analysis,age,differentiation,tumor size,Bismuth-Corlette classification,portal vein invasion,lymph node metastasis,hepatic artery invasion,liver invasion,preoperative biliary drainage,resection margin and preoperative CA19-9 levels were identified as significant prognostic factors.On multivariable analysis,lymph node metastasis,resection margin and preoperative CA19-9 levels were independent prognostic factors of survival.Conclusions A raised preoperative CA19-9 level was an independent prognostic factor of survival for hilar cholangiocarcinoma.The most discriminative cutoff point of CA19-9 for prognosis was at 150 kU/L.

Objective To investigate the risk factors of spontaneous rupture of BCLC stage A and stage B hepatocellular carcinoma (HCC),and to review the surgical outcomes.Methods From April 2002 to November 2006,89 patients who suffered from spontaneous rupture of HCC of BCLC stage A and stage B were included into this study.A control group of 171 patients was selected by matching the sex,age and BCLC stage.Clinical data and survivals were collected and analysed.Results On multivariate analysis,hypertension (HR 7.38,95％CI:1.91 ～28.58,P＜0.05),cirrhosis (HR6.04,95％ CI:2.83 ～12.88,P ＜ 0.05) and tumor location in segments Ⅱ,Ⅲ,Ⅵ (HR 5.03,95％ CI:2.70 ～ 6.37,P ＜ 0.05) were predictive factors of spontaneous rupture of HCC.In the study group,the median survival and median disease-free survival were 12 months (range,1 ～ 78 months) and 4 months (range,0 ～ 78 months) respectively.The overall survival rates and disease-free survival rates at 1-,3-and 5-year were 66.3％,23.4％,10.1％ and 57.0％,16.8％,4.5％,respectively.Only radical resection remained predictive of overall survival (HR 0.32,95％ CI:0.08 ～ 0.61,P ＜ 0.05) and disease-free survival (HR 0.12,95％ CI:0.01 ～ 0.73,P ＜ 0.05).Conclusions Tumor location,as well as hypertension and cirrhosis were associated with spontaneous rupture of HCC.One-stage hepatic resection should be recommended to patients with ruptured HCC of BCLC stage A and stage B.