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Objective To find out whether NOTCH receptors can serve as direct downstream targets of transforming growth factor β(TGF-β)/Smad4 signaling in endothelial cells.Methods Real-time PCR and Western blotting were performed to verify whether the expression of notch1 and notch4 was regulated by TGF-β pathway.Luciferase reporter assay was employed to investigate how the promoter of notch1 and notch4 was regulated by TGF-β.Then, ChIP assay was used confirm whether the promoter of notch1 and notch4 physically interacted with SMAD protein.Results TGF-β1 and bone morphogenetic protein 4 (BMP4) treatment increased the expression of both notch1 and notch4 at the transcriptional level.In addition, SMAD4 was physically associated with the SMAD binding sites on the notch4 promoter, which was largely enhanced under the treatment of TGF-β1 and BMP4.Importantly, TGF-β1 and BMP4 failed to transactivate notch4 in the absence of endogenous SMAD4 or the SMAD binding regions on the notch4 promoter.Conclusion The expression of NOTCH receptor can be directly up-regulated by SMAD4-mediated TGF-β/BMP signaling in cerebrovascular endothelial cells.
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Objective To investigate the protective effect of fibrauretin injection against acute lung injury induced by lipopolysaccharide (LPS) in mice.Methods Seventy-two healthy male adult Kunming mice were randomly divided into 6 goups:normal control group,LPS group (intratracheal instilation of 5 mg/kg of LPS),hydrocortisone group (intraperitoneal injection of 3.3 mg/kg of hydrocortisone,once daily for 3 days,and intratracheal instilation of 5 mg/kg of LPS on the 4th day) and low dose,medium dose and high dose of fibrauretin groups (intraperitoneal injection of 2,10 and 50 mg/kg of fibrauretin,respectively,once daily for 3 days,and intratracheal instilation of 5 mg/kg of LPS on the 4th day).The mice in each group were sacrificed by dislocation 24 h after intratracheal instilation of LPS.The lung tissues of partial mice in each group were extracted and weighed to calculate the lung coefficients,and bronchoalveolar lavage (BAL) was performed in partial mice in each group to collect the BALF for counting the inflammatory cells.Results The lung cofficients of mice in LPS group were significantly higher than those in the normal control group (P＜0.05).The lung cofficients of mice in hydrocortisone group,low dose,medium dose and high dose of fibrauretin groups were significantly lower than those in LPS group (P ＜ 0.05).For the percentage of inflammatory cells in BALF and the percentage of neutrophils in inflammatory cells,LPS group was significantly higher than the normal control group (P＜0.01),and compared with LPS group,hydrocortisone group,low dose,medium dose and high dose of fibrauretin groups were significantly decreased (P＜0.01).Conclusion Fibrauretin injection can significantly ameliorate the inflammatory reaction degree in mice with acute lung injury induced by LPS.
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Objective To analyze the composition and distribution of pathogens from 1366 superficial candidiasis cases in Shanghai.Methods Candida species identification was carried out for 1366 adults or children with superficial candidiasis by using CHROMagar Candida plates,API20C AUX system,etc.Pal's agar,Xylose assimilation and the test for growth at 45 ℃ were utilized to differentiate Candida dubliniensis.Newly identified pathogenic Candida species including Candida orthopsilosis,Candida metapsilosis,Candida fermentati,Candida nivariensis and Candida bracarensis were differentiated by molecular biological methods.Finally,the composition and distribution of pathogens in superficial candidiasis cases were statistically analyzed.Results A total of 1366 Candida strains,included 2 Candida orthopsilosis strains and 4 Candida metapsilosis strains,were isolated from these cases.Among these isolates,Candida albicans predominated(79.0％),followed by Candida parapsilosis(9.5％),Candida tropicalis(2.9％)and Candida guilliermondii(1.9％).The composition of Candida species was significantly different between child and adult patients(x2 =196.46,P ＜ 0.01),with the isolation rate of non-albicans Candida species being 14.4％ and 45.8％ respectively in child and adult patients.Conclusions Candida albicans is still the dominant pathogen of superficial candidiasis.Candida orthopsilosis and Candida metapsilosis can cause superficial candidiasis.The isolation rote of non-albicans Candida species is higher in adult patients than in child patients.
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Objective To discuss whether mild hyperuricemia can lead to kidney damage and the protection of decreased uric acid,through observing that hyperuricemia did damage to glomerulus endothelial function and cell proliferation of vascular smooth muscle in rats. Methods Fifty-four male SD rats were divided into four groups,the control group,model group (Oxonate),allopurinol group and Oxonate+allopurinol group.Rats were administered on a low sodium diet and their systolic blood pressure (SBP) were measured each 10 days.ELISA was used to detect rat plasma markers of endothelial function damage [nitric oxide (NO),type-1 plasminogen activator inhibitor (PAI-1),endothelin 1 (ET-1)] and cell proliferation of vascular smooth muscle[plateletderived growth factor (PDGF),cycloxygenase 2 (COX2),monocyte chemotactic protein-1 (MCP-1)],and the markers of inflammatory reaction[interleukin-18 (IL-18),tumor necrosis factor α(TNF-α)].PDGF and nitric oxide synthase (NOS) levels of rats were detected by immunohistochemical method.Renal tissue pathology of rats was observed. Results Compared to the control group,the plasmic concentration of COX2,ET-1,IL-18,PAI-1,PDGF,TNF-o,MCP-1 increased,and NO decreased significantly in rats of model group (all P＜0.05),expression of NOS significantly reduced and PDGF increased (all P＜0.05).Under light microscope,vascular wall thickening,intimal proliferation and lumen slight stricture without uric acid crystals in renal tissue were found in model group,which were obviously improved by using allopurinol. Conclusion Mild hyperuricemia can do damage to endothelial function of glomerulus and lead to vascular cell proliferation,which can be improved through decreasing uric acid.
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Objective To evaluate the role of Restriction fragment length polymorphism (RFLP) analysis in detection of the fragment of GEF1α/a gene which are both located at ct and a mating type loci in identification of species, varieties, genotypes and mating types of Cryptococcus neoformans species complex(Cryptococcus neoformans and Cryptococcus gattii). Methods The GEF1α/a gene was selected from 20 genes which both located at α and a mating type loci for RFLP analysis, according to the requirements of sequence similarities and primer design in PCR-RFLP analysis. Primer pair was designed from the conserved regions of GEF1α/a genes of distinct genotypes and mating types of reference strains to amplify a fragment of GEF1α/a gene from Cryptococcus neoformans and Cryptococcns gattii strains tested. Sequence alignment,restriction maps analysis, endonucleases selection and electrophoresis stimulation were conducted by using DNAMAN and Vector NTI software. EeoT14 Ⅰ and Hap Ⅱ endonucleases were selected for RFLP analysis of the GEF1α/a fragments amplified from 125 isolates of Cryptococcns neoformans and Cryptococcus gattii. Results An approximate 1 300 bp fragment was amplified from total 82 Cryptococcus neoformans and 43 Cryptoceccus gattii isolates. However, negative PCR results were found in the reference strains of Cryptococcus laurentii, Candida albicans, Candida tropicalis, Candida parapsilosis, Candida krnsei,Candida glabrata, Trichosporon asahii, Aspergillus fumigatns and Aspergillus flavus. RFLP analysis successfully identified the species, varieties, genotypes and mating types of total 125 isolates of Cryptococcus neoformans and Cryptococcns gattii tested in this study. Condusion PCR-RFLP analysis of the GEF1α/a fragment has the potential value in identification of species, varieties, genotypes and mating types of Cryptococeus neoformans species complex simultaneously and rapidly, and may be a useful tool in molecular epidemiological analysis.