Protective effect of aucubinon on the eucommia ulmoides on skin photoaging induced by UVB / 国际中医中药杂志

Objective To explore the protective effect of aucubin on the photoageing of the skin that induced by ultraviolet (UVB) radiation. Methods Human skin fibroblasts were divided into normal group, model group and high, medium and low dose groups of apocynin by random method. Except the normal group, the other groups of cells were subjected to UVB radiation with a radiation wavelength of 311 nm, irradiation intensity of 9.00 mw/cm2, radiation time of 2.8 s, radiation dose of 25 mJ/cm2. The cells in the high, medium and low doses of aglycone were cultured for 24 h at an equal volume of 400, 200, 100 μg/ml, and then subjected to UVB irradiation, and then cultured for 24 h. The normal group and the model group were cultured for 48 h in a common medium. The apoptosis rate of each group was detected by flow cytometry. The levels of IL-1, IL-6 and TNF-α in each group were detected by ELISA. The expressions of Bax and Bcl-2 mRNA in each group were detected by RT-PCR. Results Compared with the model group, the apoptosis rate (4.84% ± 0.24%, 12.32% ± 0.67% vs. 35.63% ± 2.77%) in the medium and high dose group significantly decreased (P<0.05), the content of IL-1 (21.14 ± 1.94 ng/ml, 16.17 ± 0.74 ng/ml vs. 22.55 ± 1.02 ng/ml), IL-6 (17.46 ± 0.93 ng/ml, 14.51 ± 0.79 ng/ml vs. 18.39 ± 2.05 ng/ml), TNF-α (44.21 ± 1.16 ng/ml, 35.94 ± 3.08 ng/ml vs. 45.67 ± 2.28 ng/ml) significantly increased (P<0.05). Compared with the model group, the expression of Bax mRNA (0.29 ± 0.05, 0.42 ± 0.05, 0.51 ± 0.04 vs. 0.59 ± 0.05) significantly decreased, and the expression of Bcl-2 mRNA (0.52 ± 0.04, 0.44 ± 0.03, 0.35 ± 0.03 vs. 0.26 ± 0.04) significantly increased (P<0.05). Conclusions The aucubin has protective function to the damage of skin fibroblasts which is induced by UVB radiation, indicating the protective function of decreasing Bax mRNA expression, increasing Bcl-2 mRNA expression, reducing the expression of inflammatory factor IL-1, IL-6, TNF-α, holding-up the cell apoptosis which is induced by UVB radiation, and inhibiting apoptosis of cell that caused by UVB radiation, playing an role of protective fibroblasts for delay of the photoageing.

Effect of siRNAs on HSV-1 Plaque Formation and Relative Expression Levels of RR mRNA / 中国病毒学·英文版

RNA interference(RNAi)is a process by which introduced small interfering RNA(siRNA)can cause the specific degradation of mRNA with identical sequences. The human herpes simplex virus type 1(HSV-1)RR is composed of two distinct homodimeric subunits encoded by UL39 and UL40, respectively. In this study, we applied siRNAs targeting the UL39 and UL40 genes of HSV-1. We showed that synthetic siRNA silenced effectively and specifically UL39 and UL40 mRNA expression and inhibited HSV-1 replication. Our work offers new possibilities for RNAi as a genetic tool for inhibition of HSV-1 replication.

In vitro Anti-viral Activity of the Total Alkaloids from Tripterygium hypoglaucum against Herpes Simplex Virus Type 1 / 中国病毒学·英文版

Herpes simplex virus type 1 (HSV-1) is a commonly occurring human pathogen worldwide. There is an urgent need to discover and develop new alternative agents for the management of HSV-1 infection. Tripterygium hypoglaucum (level) Hutch (Celastraceae) is a traditional Chinese medicine plant with many pharmacological activities such as anti-inflammation, anti-tumor and antifertility. The usual medicinal part is the roots which contain about a 1% yield of alkaloids. A crude total alkaloids extract was prepared from the roots of T. hypoglaucum amd its antiviral activity against HSV-1 in Vero cells was evaluated by cytopathic effect (CPE) assay, plaque reduction assay and by RT-PCR analysis. The alkaloids extract presented low cytotoxicity (CC_(50) = 46.6 μg/mL) and potent CPE inhibition activity, the 50% inhibitory concentration (IC_(50)) was 6.5 μg/mL, noticeably lower than that of Acyclovir (15.4 μg /mL). Plaque formation was significantly reduced by the alkaloids extract at concentrations of 6.25 μg/mL to 12.5 μg/mL, the plaque reduction ratio reached 55% to 75% which was 35% higher than that of Acyclovir at the same concentration. RT-PCR analysis showed that, the transcription of two important delayed early genes UL30 and UL39, and a late gene US6 of HSV-1 genome all were suppressed by the alkaloids extract, the expression inhibiting efficacy compared to the control was 74.6% (UL30), 70.9% (UL39) and 62.6% (US6) respectively at the working concentration of 12.5μg/mL. The above results suggest a potent anti-HSV-1 activity of the alkaloids extract in vitro.

Effect of basic fibroblast growth factor and insulin on the proliferation and differentiation of mouse chondrocytes / 中国组织工程研究

BACKGROUND: Based on the characteristics of cartilage tissue, such as consisting of single type of cells, the cartilage cells or chondrocyte, absence of blood vessel, rather low consumption level of oxygen and nutrition, low level of allo-immunocompetence and simple function in vivo, it seems to be easy for cartilage cell lines to be established for tissue and cell transplantation. We want to set up a cell line with the purpose of current use in tissue engineering in vitro. It will provide the basis for artificial tissue and organ that will become to be standardized and yielded in batch.OBJECTIVE: To explore the potential stimulatory effects of basic fibroblast growth factor(bFGF) and insulin on the proliferation and differentiation in primary culture mice chondrocytes in vitro. The effect and application of the cell factors will be evaluated for tissue engineering.DESIGN: A grouping controlled and repeated trial was conducted with the cells as the subjects.SETTING: Key laboratory of tissue transplantation and immunology of a college.MATERIAIS: The experiment was completed in the Key Laboratory of Tissue Transplantation and Immunology of the Ministry of Education, Jinan University from November 2002 to May 2003. Cultured cartilage cells at random were obtained as the study objects.METHODS: Mice cartilage cells were cultured in medium at the minimum concentrations of serum. The effects of different concentration of bFGF and insulin on the proliferation and differentiation in mice cartilage cells were observed with WST1 and immunofluorescence staining.MAIN OUTCOME MEASURES: Primary results: ① Effect of bFGF on proliferation of primary cultured mice cartilage cells. ② Effect of insulin on proliferation of primary cultured mice cartilage cells. Secondary results:morphological observation of cartilage cells RESULTS: Primary cultured mice cartilage cells were cultured in medium at the minimum concentration of serum(4 g/L fatal bovine serum). It was found that bFGF and insulin might play an important role on the proliferation and growth of mice cartilage cells in a dose-dependent manner. In addition, morphological observation of cartilage cells showed that both bFGF and insulin not only promoted the proliferation of the cells but also enhanced the matrix secretion of cartilage cells.CONCLUSION: Both bFGF and insulin can stimulate the proliferation of cartilage cells in vitro.

Possibility of 3T3 fibroblast growth on blood fibrin clot in culture medium with recombinant human basic fibroblast growth factor / 中国病理生理杂志

AIM: To investigate the possibility of 3T3 fibroblast growth on blood fibrin clot in culture medium with recombinant human basic fibroblast growth factor (rhbFGF). METHOD: Growth of the cells on blood fibrin clot was studied by phase-contrast, scanning and transmission electron microscopy and by Giemsa stain and MTT assay. RESULTS: The optimal concentration of rhbFGF for proliferation and survival of the cells was 100 ng/mL. The cells also grew on blood fibrin clot scaffold in the low-serum medium containing 100 ng/mL rhbFGF, and a greater number of the cells survived after 48 hours incubation compared to that after 24 hours. The elongated filopodia appeared to bridge the gaps among the fibroblasts after 24 hours incubation. Further incubation to 72 hours, a greater number of platycytes were found to be joined together by lamellopodia. CONCLUSION: 3T3 fibroblasts could grow and survive on blood fibrin clot in the low-serum medium containing rhbFGF, and a combination of blood fibrin clot and rhbFGF may have over proportional effects on wound healing.

Effects of bFGF and insulin on proliferation of mice cartilage tissue cells / 中国病理生理杂志

AIM: To explore the effects of basic fibroblast growth factor ( bFGF) and insulin on the cell proliferation and differentiation in primary cartilage cells. METHODS: After induction with different concentrations of bFGF and insulin, cell proliferation was measured with WST-1 method and fluoroscope methods. RESULTS: bFGF and insulin exerted their related action on primary cartilage cells in 0.4% fatal bovine serum at different concentrations. 25 ?g/L bFGF and 5 mg/L insulin promoted cell proliferation significantly. CONCLUSION: bFGF and insulin prolong the survival time and promote cell proliferation in primary cartilage cells.

Construction of the retroviral vector recombinant of HBV-S gene and its expression in antigen presenting cells / 中国病理生理杂志

AIM: To investigate the effectiveness of recombinanted retrovirus vector in gene therapy. METHODS: The retroviral vector pLXSN S was constructed and transferred into PA317 by means of electroporation, then HepG 2?RAW264.7 and EL 4 cells were infected with the pseudovirus produced from PA317, which highly expressed HBsAg . HBsAg expression was tested by RT PCR and ELISA. RESULTS: HBsAg was expressed variously in the eukaryotic cells mentioned above. HBsAg ( A value) of the cell supernatants (48 hours) were 0.92,0.53,0.42, respectively. CONCLUSION: The vector used in this study was an effective vector to carry genes of interest to target cells and macrophage, and high level HBsAg was expressed in antigen presenting cell such as macrophage, It indicated that plasmid immunity can induce the B lymphocyte and T lymphocyte response to hepatitis B virus surface antigen by stimulating macrophage. As a vector, it may be useful in the test for gene immunity and gene therapy.