The preliminary research on the gap junction mechanisms for synergistic effects of liuwei di-huang pill containing serum on suicide gene therapy of melanoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the gap junction mechanisms for synergistic effects of liuwei di-huang pill (LDP) containing serum on HSV-tk/GCV suicide gene therapy of mouse malignant melanoma B16 cells.</p><p><b>METHODS</b>The LDP containing serum (2.5% LDP serum group, 5.0% LDP serum group, and 10.0% LDP serum group) and the control serum group were set up. The effects of LDP on mRNA expressions of Cx26 and Cx43 in mouse malignant melanoma B16 cells were detected by RT-PCR assay. The effects of LDP on protein expressions of Cx26 and Cx43 in B16 cells were detected using Western blot and indirect immunofluorescence assay. The interference efficiencies of Cx26-309, Cx26-337, Cx26-367, Cx26-2098 SiRNA on Cx26 gene in B16 cells were detected using RT-RCR technique. Cx26-2098 SiRNA, due to the optimal interference efficiency, was chosen to interfere Cx26 gene, and the bystander effect of LDP + HSV-tk/GCV was observed. The effects of the gap junctional intercellular communication (GJIC) inhibitor glycyrrhetinic acid on the killing of LDP + HSV-tk/GCV system to B16 cells were detected by MTT assay. In this experiment, the control serum group, 2.5% LDP serum group, 5. 0% LDP serum group, 10.0% LDP serum group, the control serum combined GCV group, 2.5% LDP serum combined GCV group, 5.0% LDP serum combined GCV group, 10.0% LDP serum combined GCV group, 2.5% LDP serum combined GCV + glycyrrhetinic acid group, 5.0% LDP serum combined GCV + glycyrrhetinic acid group, 10.0% LDP serum combined GCV + glycyrrhetinic acid group were set up. The final concentration of GCV was 20 micromol/L. The final concentration of glycyrrhetinic acid was 50 micromol/L.</p><p><b>RESULTS</b>LDP containing serum could increase the protein and mRNA expressions of Cx43 in B16 cell in a dose-dependent manner. It had bi-directional regulation on the Cx26 protein and mRNA expressions. The low dose LDP had inhibition while high dose LDP could up-regulate its expression. After SiRNA interfered Cx26 gene, there was no obvious change in the bystander effect of LDP combined suicide gene therapy between before and after interference. There was significant reduction in the inhibition rate between before (48.75%, 59.39%, and 69.28%) and after blockage (29.14%, 38.71%, and 58.13%) of glycyrrhetinic acid in the 2. 5%, 5.0%, 10.0% LDP serum combined GCV groups (P <0.01).</p><p><b>CONCLUSION</b>The synergistic effects of LDP containing serum on HSV-tk/GCV suicide gene therapy in mouse malignant melanoma B16 cells were correlated with the gap junction mechanisms, which might be achieved through increasing the expressions of Cx26 and Cx43.</p>

Isolation and Identification of Thermoacidiphilic Bacteria from Apple Juice Concentrate / 微生物学通报

The thermotolerant bacteria which was isolated from the Apple Juice Concentrate(AJC) was investigated by comparison with the standard strain of Aliyclobacillus acidoterrestris DSM3922 using Kirin-medium acidified with malic acid.The results shows the two polluted bacteria isolated from AJC can grow under the temperature of 21?C～55?C and pH of 2.4～6.2,which was corresponded with the characteristics of the thermo-acidiphilic Alicyclobacillus spp..Further more,the cell morphology,colony morphology,cultural characteristics and physiological characteristics tests indicated the two isolated strains of thermotolerant bacteria have obviously similar characteristics with the standard strain of Aliyclobacillus acidoterrestris DSM3922.