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Objective To investigate the factors that influence the fermentation process of compound Chinese medicine and determining the optimum fermentation with single factor experiment and response surface methodology. Methods Through controlling the factors in the fermentation process of compound Chinese medicine (such as fermentation bacteria, fermentation time, fermentation temperature, inoculum amount, etc.), with its increase rate of total peak area as evaluation indicator, the alcohol extracts before and after fermentation were monitored and comparative evaluated by HPLC, and the optimum fermentation process was determined by response surface methodology. Results The fermentation process optimized by single factor experiment and response surface methodology was as follows:SZ-2 strain served as the fermentation bacteria, temperature was 33 ℃, inoculum amount was 4%, and time was 3.5 d, the average increase rate of total peak area was 31.24%. Conclusion HPLC can be used to identify and evaluate the fermentation of compound Chinese medicine under the different factors, and to clarify the optimal fermentation process by response surface methodology, which provide reference for the development of fermentation process.
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Objective To investigate the effects of glucocorticoids o n the expression of Th1/Th2 cytokines at mRNA and protein levels in peripheral b lood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SL E). Methods Th1 cytokine (IFN-IL-10) in PBMCs fro m 48 patients with SLE were detected before and after treatment with glucocortic oids by RT-PCR for mRNA expression and ELISA for protein production. The systemi c lupus erythematosus activity measure (SLAM) scores of patients were compared b efore and after treatment. Results After treatment, the expression and product ion of IFN- versus 1.4094 pg/mL; P
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Objective To investigate the effects of siRNA targeting CD 147 gene on the expression of CD147 in melanoma cell line A375, and on proliferation of these cells. Methods Previously prepared recombinant plasmid pSUPER/CD147 siRNA was used. In this study, the recombinant was transfected into the A375 cells. The mRNA expression of CD147 was measured by semi-quantitive RT-PCR, the proliferation of the cells by MTT assay. Results After the transfection with pSUPER/CD147 siRNA1 and pSU-PER/CD147 siRNA2, the mRNA expression of CD147 in the A375 cells was significantly down-regulated by 90.81% (P
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Objective To investigate the expression and role of CD147in the differentiation of nor-mal keratinocytes and squamous cell carcinoma(SCC)cells.Methods CD147expression was analyzed immunohistochemically in20cases of verruca vulgaris(VV),various benign,premalignant and malignant epidermal tumors including21cases of seborrheic keratosis(SK),20actinic keratosis(AK),20Bowen' s disease(BD)and57squamous cell carcinoma.SCCs were classified using Broder's system of grading.Im-munoblot analysis was used to observe CD147expression in normal keratinocyte(HaCaT)and SCC cell(HSC-5)in vitro during their differentiation process induced by calcium.The effects of high-calcium medi-um culture and CD147antibody on the differentiation-related morphology of HaCaT and HSC-5cells were also observed.Results The same staining pattern of CD147was shown in20VV and21SK specimens as in normal epidermis.Positive CD147staining throughout the lesion was shown in4of20AK and7of20BD specimens.Positive CD147staining at the periphery of tumor nests was shown in8of20gradeⅠSCC specimens.CD147expressed throughout tumor nests in16of20gradeⅡSCC and all of the17gradeⅢSCC specimens.Immunoblot analysis revealed decreased CD147expression in both HaCaT and HSC-5cells during differentiation process induced by calcium.Not only high-calcium medium but also CD147antibody could induce differentiation-related morphology of both HaCaT and HSC-5cells in vitro.Conclusion These results suggest that CD147is a novel low-differentiation marker of keratinocyte,which might inhibit the dif-ferentiation of both normal keratinocyte and SCC cell.
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Objective To examine the cellular distribution and the role of a-catenin in epidermal tumors. Methods Expression of a-catenin was investigate d by immunohistochemical staining in 20 patients with Bowen′s disease (BD), 20 squamous cell carcinoma (SCC), 20 basal cell carcinoma (BCC), and 40 normal cont rols. Results Expression of a-catenin was strongest on the cell membranes of b asal cells, but nearly negative in the cytoplasm and the nuclei of epidermal cel ls in normal controls. Expression of a-catenin was significantly lower on the cell membranes in SCC and BCC cells than that in normal epidermal cells (P