To investigate the role of ERC,6 gene in the growth rate and antifungal susceptibility,Aspergillus fumigatus strain with extra copies of ERG6 gene was constructed.Open reading frame (ORF) of putative ERG6 gene was searched in A.fumigatus genome.A PCR fragment, ERG6 ORF sandwiched by its flanking sequences (about 1 kb respectively),was amplified and was then subcloned into vector pRG-AMA1- NotI to produce a recombinant plasmid pERG6,which was further transformed into uracil auxotroph A.fumigatus strain AF293.1 to produce the transformant AF-pERG6.In the same time,empty plasmid pRG-AMA1-NotI was also transformed into A.fumigatus strain AF293.1 to produce the transformant AF-empty as a control.Radial growth of the transformants was tested on minimal medium (MM) and YAG medi- um.Antifungal susceptibilities of these resahing transformants,AF-pERG6 and AF-empty,to the common antifungal agents were performed by using both disk diffusion and broth microdillution methods.A.fumigatus genome contains a ERG6 gene,of which the ORF size is 1 256 bp.Comparing to Candida albicans and Sacchromyces cerivisiae sterol methyltransferase,A.fumigatus Erg6p had 57% and 50% identity, and had 70% and 63% similarity in amino acid sequences,respectively.Radial growth of transformant AF-pERG6 was slower than that of transformant AF-empty.The antifungal susceptibilties of transformant AF-pERG6 to the antifungal drugs itraconazole,voriconazole,terbin- aline,amphotericin B,caspofungin and grisefolvin were same to that of transformant AF-empty.In A.fumigatus,extra copies of ERG6 gene have no effect on antifungal susceptibilities to itraconazole,voriconazole,terbinafine,amphotericin B,caspofungin and grisefolvin.Radial growth of A.fumigatus harboring extra copies of ERG6 gene becomes slower compared to the control.

In recent years,with the increasing of immunocompromised patients in clinical practice,invasive candidiasis has been increased accordingly. Although several antifungal drugs,including polyenes,azoles,5-flucytosine,and echinocandins,have been used widely to treat invasive candidiasis,the treatment failure that caused by the resistance of Candida albicans to these antifungal drugs have been reported widely. Here,we reviewed the molecular mechanisms of resistance to azoles,polyenes,5-flucytosine,and echinocandins,such that to provide an update on these issues.

Animals ; Rats ; Specimen Handling ; methods ; Spinal Cord

Adult ; Female ; Humans ; Hypopituitarism ; etiology ; Otorhinolaryngologic Surgical Procedures ; adverse effects ; Postoperative Period ; Turbinates ; surgery

Objective To explore the differences in health care quality,scale,output,efficiency,cost and financial condition between freestanding secondary hospitals and system-affiliated secondary hospitals in Shanghai,and analyse the determinants of hospital integration. Methods Eleven upper secondary hospitals in Shanghai integrated between 2000 and 2004 were selected,and another 40 secondary hospitals (including 30 upper secondary hospitals and 10 middle secondary hospitals) without integration were served as controls. Using related data of 1999,Mann-Whitney U test was performed to analyse the differences between these two groups,and Logistic regression analysis was conducted to explore the determinants of hospital integration. Results There were significant differences in health care quality,scale,and output between these two groups (P0.05). It was revealed by Logistic regression analysis that health care quality,scale,output,and financial condition were determinants of integration. Conclusion System-affiliated secondary hospitals have advantages over freestanding hospitals in health care quality,scale,output and financial condition,and those with better health care quality,larger scale,larger output and better financial condition are more likely to be integrated by tertiary hospitals.

Objective To evaluate the susceptibility of itraconazole-resistant Aspergillus fumigatus isolates to four other antifungal drugs.Methods Six strains of Aspergillus fumigatus were serially isolated from a patient with lung aspergilloma,who was resistant to itraconazole therapy.Clinical Laboratory Stan- dard Institute broth microdilution method M38-A and E-test were applied to test the susceptibility of A.fumi- gatus strains to amphotericin B,itraconazole,voriconazole,caspofungin and micafungin.Results With M38-A method,the minimal inhibitory concentration(MIC)of itraconazole was 0.5?g/mL for 2 strains and more than 16?g/mL for the other 4 strains;the MICs of amphotericin B and voriconazole were 1?g/mL and 0.25-1?g/mL respectively for all the 6 strains;and the minimal effective concentration(MEC)of caspofungin and micafungin were no more than 0.03?g/mL for all the 6 strains.Moreover,E-test results also revealed that caspofungin and voriconazole had a good antifungal activity against all the strains,including the 4 itraconazole-resistant ones.Conclusion The itraconazole-resistant A.fumigatus strains isolated from a patient with lung aspergilloma are susceptible to amphotericin B,voriconazole,caspofungin,and micafungin.

ObjectiveTo identify serum specific protein biomarkers for the early diagnosis of lymphoma patients by using surface enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF-MS)protein chip array.MethodsSELDI-TOF-MS technique and weak cation exchanger (WCX2)protein chip were used to detect the serum proteomic patterns of 96 patients with lymphoma (86 patients with non-Hodgkin lymphoma and 10 patients with Hodgkin lymphoma)and 30 normal control.The serum level of LDH were measured by biochemical methods,the serum level of β2-MG and CA125 were detected by enzyme-linked immunosorbent assay (ELISA).ResultsFour protein peaks pattern mass/charge ratio (M/Z) 6880,8564,8692 and 13751 were decreased in lymphoma patients,and they could be used to distinguish lymphoma from healthy people and other malignant tumors.Their sensitivity and specificity were 82.29 ％ and 80.00 ％,78.13 ％ and 73.30 ％,75.00 ％ and 80.00 ％,71.88 ％ and 83.30 ％ respectively in lymphoma patients.When it was used to early diagnosing lymphoma in stage Ⅰ or Ⅱ,the sensitivity of four protein peaks pattern (M/Z 6880,8564,8692 and 13 751) were 77.55 ％,71.43 ％,71.43 ％ and 67.35 ％respectively and the specificity of all four protein peaks pattern were 100.00 ％.Four protein peaks pattern sensitivity were higher than LDH,CA125 and β2-MG respectively.ConclusionApplication of SELDI-TOF-MS can be of great potential for the early diagnosis of lymphoma patients.

OBJECTIVETo explore the relationship between the expression of heat shock protein 90, 60 and 27 (HSP90, HSP60 and HSP27) and genetic damage in peripheral blood of workers exposed to coke oven emissions.

METHODS288 coke oven workers in a steel factory were divided into the high-dose group and the low-dose group on the basis of environment monitoring result and work place. There were 172 men in high-dose group (workers who worked at the oven top and oven side) and 116 men in low-dose group (workers who worked at the oven bottom and others who were engaged to aided work). 38 workers unexposed occupationally to carcinogenic substances were selected as the control group, who were employed in medical therapy unit nearby 2 kilometers from the steel factory. Their general information, history of personal and occupational exposure, and the work environment were investigated. Blood samples were collected immediately after a shift at the end of a working day from 288 coke oven workers and 38 control workers. Levels of HSP90, HSP60 and HSP27 in peripheral blood lymphocytes were measured by Western blot, and the degree of DNA damage was detected by the comet assay.

RESULTSLevels of HSP90 in peripheral blood lymphocytes in three groups were 0.24 +/- 0.32, 0.12 +/- 0.30 and 0.06 +/- 0.33 respectively. They increased significantly compared with that of the control. But levels of HSP60 and HSP27 were not significantly different among those groups. Compared with the control group, there was significant difference in tail length, olive tail moment et al of SCGE (G +/- s(G)) of occupational exposure workers. High-dose group > low-dose group > control group (P < 0.05). The degree of DNA damage increased with the rise of exposure BaP dose (Spearman r = -0.345, P < 0.01).

CONCLUSIONLevels of HSP90 in peripheral blood lymphocytes and the degree of DNA damage increase with the rise of exposure polycyclic aromatic hydrocarbons (PAHs) dose.

Adult ; Chaperonin 60 ; blood ; Coke ; adverse effects ; Comet Assay ; DNA Damage ; drug effects ; HSP27 Heat-Shock Proteins ; blood ; HSP90 Heat-Shock Proteins ; blood ; Humans ; Lymphocytes ; metabolism ; Male ; Occupational Exposure ; adverse effects ; Polycyclic Aromatic Hydrocarbons ; adverse effects

To ascertain current situation of wild Marsdenia tenacissima resources in Honghe, Yunnan province, the distribution, habitat characteristic and resources reserves of M. tenacissima were surveyed based on interviews and investigation. The results showed that M. tenacissima was found in 7 counties such as Jinping, Mengzi etc, and distributed mainly on the mountainsides from 800 m to 1 200 m. And distribution was affected by many factors, such as light, heat, topography, soil, and vegetation. M. tenacissima grew well in distribution areas. M. tenacissima had averagely a weight of 2.8 kg per plant. Resources reserve of M. tenacissima in Honghe was estimated to 1 300 tons by now but it reduced rapidly in resent years, the wild resources reserve may not meet demand of market. Resources protection and wildlife tending would be conducted to deal with increasing medication requirements.
China ; Ecosystem ; Marsdenia ; classification ; growth & development ; Plants, Medicinal ; classification ; growth & development ; Soil ; chemistry

Objective To screen the antibacterial activity of Chinese traditional medicines against Yersinia pestis.Methods Six Chinese traditional medicines(Coptis Chinesis etc)were selected and extracted with pure water to make a concentration of 1 mg/L.Yersinia pestis strain 201 and EV 76 were used to determine the minimal inhibitory concentrations(MIC)of these selected medicines in vitro with liquid dilution method.Results Three herbs had inhibition effects on the strain 201 and EV76 in different extents,among which Rheum palmatum had the strongest effect and MIC was 0.025 00 mg/L.Furthermore,the Chinese traditional medicine had the same MIC on both strain 201 and EV76.Conclusions Chinese traditional medicines commonly used have inhibiting effect on Yersinia pesti.