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The correlated proteome of Penicillium marneffei resistant to fluconazole was investigated in the present study, in which 11 strains of P. marneffei of both the mycelial and yeast forms sensitive to fluconazole were cultivated in Sabouraud's liquid medium containing 8 g fluconazole and the MICs of fluoconazile to P.marneffei before and after inducing cultures were tested by E-test method. The strains of the mycelial and yeast forms showing the most significant increase in MIC value were selected and the protein clip CM10 was used to detect the proteome differences before and after inducing cultures. It was found that 11 strains of the mycelial forms and 2 strains of the yeast forms could tolerate the action of fluconazole and could grow at a drug concentration of 8 μg/mL. Furthermore, the MICs of fluoconazole to the mycelial forms were significantly increased after 7 days of inducing culture and the geometric means of MICs were increased from 1.22 μg /mL to 55.56 μg/mL. After the mycelial forms had been induced to be resistant, 16 proteins were highly expressed with specific expression of the 4581.1 Da and 6109.7 Da proteins; whereas 22 resistant yeast form were highly expressed with specific expression of the 3575.2 Da, 8507.0 Da and 8563.3 Da proteins. These results suggest that the mycelial form of P.narneffei seems to be more tolerant to the action of fluconazole than the yeast form. The resistance of these organisms to fluconazole may be associated to some specific proteins.
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Objective To study the effect of NS398, an inhibitor of cyclooxygenase 2 (COX2), on the growth and apoptosis of human squamous cell carcinoma cell line Tca8113. Methods Cultured Tca8113 cells were incubated with NS398 (0, 6.25, 12.5, 25, 50, 100 μmol/L) for 24, 48 and 72 hours, respectively. Thereafter, MTT method, flow cytometry and transmission electron microscopy were applied to detect the proliferation, cell cycle and apoptosis of Tca8113 cells, respectively. Results The proliferation of Tca8113 cells was inhibited by NS398 in a dose- and time-dependent manner (both P<0.05). FCM analysis showed the appearance of a typical hypodiploid apoptotic (Sub-G1) peak, an increase in the percentage of cells at G0/G1 phase and a decrease in that at S and G2/M phases in NS398 ( 100 μmol/L) -treated Tca8113 cells. Moreover, the cell proliferation index was significantly downregulated by NS398 of 100 μmol/L from 41.03 to 24.33 (P<0.05). Under an electron microscope, morphological changes characteristic of apoptosis were observed in NS398-treated Tca8113 cells. Conclusion NS398, an inhibitor of COX2, could effectively inhibit the growth of Tca8113 cells in vitro by induction of apoptosis.
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To investigate the gene mutation in a pedigree with X-linked ichthyosis (XLI) and to explore the relationship between the mutation and its clinical manifestations, genomic DNA of affected members, the normal member of the pedigree and 50 unrelated normal members was extracted with a whole blood genomic DNA extraction kit and the DNA was used as a template for the polymerase chain reaction (PCR)-mediated amplification of exon 1 and exon 10 of the STS gene. hHb6 (human hair basic keratin) gene was used as the internal control. Our results showed that the STS gene was deleted in affected members in the pedigree with X-linked ichthyosis. The normal member of the pedigree and 50 unrelated normal members had no such deletion. The proband and his mother had products in the internal control after PCR amplification. The blank control had no product. It is concluded that deletion of the STS gene existed in this pedigree with X-linked ichthyosis, and it is responsible for the unique skin lesions of X-linked ichthyosis.
Subject(s)
Gene Deletion , Ichthyosis, X-Linked/genetics , Pedigree , Steryl-Sulfatase/geneticsABSTRACT
To investigate the effects of ATRA, acitretin and tazarotene on the growth and apoptosis of human tongue squamous cell carcinoma cell line Tca8113. The effect of retinoids on growth of Tca8113 cells in vitro was examined by MTT assay and Trypan blue exclusion assay. Cell cycle analysis, early apoptosis analysis with double staining with Annexin V-FITC and PI, and active caspase-3 analysis with the staining of FITC-conjugated monoclonal rabbit anti-active caspase-3 antibody were made by flow cytometer. Streptavidin-biotin complex (SABC) immunocytochemical assays were employed for the detections of Bax/Bcl-2 proteins expressions. Our results showed that the retinoids inhibited growth of Tca8113 cells in a dose- and time-dependent manner with maximal inhibition 24 h after treatment of 10-5 mol/L. 10-5 mol/L retinoids altered cell cycle distribution of Tca8113 cells, revealing an increase in G0/G1-phase population, a decrease in S-phase population and the inhibition of G1/S switching. 10-5 mol/L retinoids significantly induced apoptosis of Tca8113 cells (all P<0.05), elevated the cells population with detectable active caspase-3 (P<0.05 for all), increased the number of cells forming Bax and decreased the number of cells forming Bcl-2 significantly (all P<0.05). Acitretin played a most prominent role among the retinoids. It is concluded that the inhibition of cell cycle progress of Tca8113 cells by ATRA, acitretin and tazarotene is one of the possible mechanisms for proliferation arrest of Tca8113 cells elicited by the retinoids. The retinoids mediate apoptosis in Tca8113 cells that may be caspase-dependent through mitochondria pathway. High concentration retinoids inhibit growth of Tca8113 cells in vitro by interfering with proliferation and inducing apoptosis of cells. Acitretin may be an alternative medicine for the prevention and treatment of tongue squamous cell carcinoma.
ABSTRACT
To investigate the gene mutation in a pedigree with X-linked ichthyosis (XLI) and to explore the relationship between the mutation and its clinical manifestations, genomic DNA of affected members, the normal member of the pedigree and 50 unrelated normal members was extracted with a whole blood genomic DNA extraction kit and the DNA was used as a template for the polymerase chain reaction (PCR)-mediated amplification of exon 1 and exon 10 of the STS gene. hHb6 (human hair basic keratin) gene was used as the internal control. Our results showed that the STS gene was deleted in affected members in the pedigree with X-linked ichthyosis. The normal member of the pedigree and 50 unrelated normal members had no such deletion. The proband and his mother had products in the internal control after PCR amplification. The blank control had no product. It is concluded that deletion of the STS gene existed in this pedigree with X-linked ichthyosis, and it is responsible for the unique skin lesions of X-linked ichthyosis.
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Objective To observe effects of survivin antisense oligonucleotide on proliferation of human malignant melanoma cell in vitro in different transfection condition.Methods By using trypan blue exclusion method,this study screened optimized transfection condition that inhibited A375 cell viability.Results ① By the phosphorothiote-modification and the liposome-encapsulation,there were significant differences in ASODN with ASODNL.ASODNL were stronger in inhibiting A375 cell proliferation(P
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Objective To investigate whether targeting survivin has the potential to sensitize A375 cells to chemotherapy.Methods By trypan blue exclusion assays we observed A375 cell proliferation inhibition induced by survivin antisense oligonucleotide(ASODN) and cisplatin.Results A375 cells treated with a combination of 5 ?mol/L ASODN and 0.5 mg/L cisplatin,and approximately 61.3% of the cells showed signs of cell death 72 h after the start of transfection trypan blue exclusion assays.Compared to treatments with either oligonucleotide or cisplatin alone,there were significant differences among the groups(P
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Objective To investigate the effects of tazarotene on the proliferation of cultured human keratinocytes and IFN-?-induced expression of HLA-DR in those cells.Methods Keratinocytes were cul-tured from normal human skin in vitro,and were treated with various concentrations of tazarotene(10 -5 ,10 -6 ,10 -7 mol/L).At24h,48h after treatment,the effects on cell proliferation were assessed by MTT method.The expression of HLA-DR was determined using immunocytochemistry techniques in cultured human ker-atinocytes incubated with tazarotene,IFN-?or both for24h.Results①The proliferation of keratinocytes was decreased when exposed to10 -7 -10 -5 of tazarotene as compared to non-exposed keratinocytes after24h and48h.Moreover,the effects on cell proliferation by tazarotene were dose-dependent;②There was rare expression of HLA-DR in normal human keratinocytes.③HLA-DR expression was inducible significantly with500u/mL of IFN-?,but failed to be induced with10 -6 mol/L of tazarotene,in keratinocytes at24h af-ter treatment.④After24h combined treatment of10 -7 -10 -5 mol/L of tazarotene and IFN-?,the induction of HLA-DR expression was significantly stronger,in a dose-dependent manner,than IFN-?alone(P
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0.05).The rate of PBMC apoptosis with CA group was noticeably increased compared to that of normal control group(P
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Objective To investigate the role of heparin binding epidermal growth factor-like growth factor(HB-EGF)in active psoriasis vulgaris.Methods HB-EGF mRNA and protein were detected by hy-bridization in situ and immunohistochemistry in normal skin tissues,lesional and non-lesional psoriatic skin of progressive stage.Results In normal skin tissues,the stain of HB-EGF mRNA and protein was located in the basal layer of epidermis(100.00%)and there was only a little expression in the suprabasal layers(16.67%).Focal overexpression was found in the suprabasal layers of non-lesional and peri-lesional psoriat-ic skin(88.00%,80.00%respectively);however,there was no HB-EGF mRNA protein expression in the superabasal layers of the central part of psoriatic lesions(0),and nearly no expression in the basal layer(4.00%).Conclusion HB-EGF may play an important role in the pathogenesis of early psoriasis.
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Objective To study the correlation between Candida strains and vulvovaginal candidiasis. Methods Two groups of Candida albicans strains were chosen. Strains of group A and group B were isolated from vaginal discharge of normal women and of patients with candidal vaginitis respectively. The two methods, AMS (automated microbiological detection and identification system) and amino black stain, were applied to detect two phenotypic parameters, biochemical reaction of the strains and secretory capacity of the proteinases respectively, and analogous analysis was performed then. Results There was a significant difference in the levels of adonitol (P
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Objective To investigate the mechanism of tazarotene in active psoriasis vulgaris. Methods HB-EGF mRNA in active psoriatic lesions before and 10 days after the treatment with tazarotene was detected by hybridization in situ. Results There was nearly no expression of HB-EGF mRNA in psoriatic lesions (9.1%); after the treatment with tazarotene, there was expression of HB-EGF not only in basal layer (95.5%), but also focal expression in suprabasal layers of epidermis (77.3%). Conclusion Tazarotene can inhibit proliferation and induce apoptosis of keratinocytes though upregulating expression of HB-EGF in psoriatic epidermis.
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Objective To explore the pathogenesis of substance P (SP) and epidermal growth factor receptor (EGFR) in early psoriasis vulgaris. Methods SP and EGFR were detected in normal skin tissues and psoriatic lesions by radioimmunoassay. Results Expression of SP was significantly enhanced in active and stable psoriasis than in recovery lesions and normal ones (P 0.05). Conclusion SP and EGFR may work together to play a key role in the early pathogenesis of psoriasis.