Preliminary Study on Effects of Solamargine on the Growth and Apoptosis of Human Hepatocarcinoma Cells HepG 2 and Its Mechanism / 中国药房

OBJECTIVE：To ex plore the effects of solamargine on the growth and apoptosis of human hepatocarcinoma cells HepG2 and its underlying mechanism. METHODS ：The effects of 0（blank group ）-12 μmol/L solamargine treatment of 24，48 h on survival rate of HepG 2 cells were investigated. The effects of 0（blank group ），6 μmol/L solamargine treatment of 10 days on cell clone formation were also investigated. The effects of 0（blank group ），4，6，8 μmol/L solamargine for 24 h on the apoptotic rate of cells，mRNA expression of Bcl- 2，Bax and caspase- 3， protein expression of Bcl- 2 and cleaved caspase- 3 as well as ratio of p-AMPKα to AMPKα were all tested. The effects of AMPK inhibitor as compound C on the protein expression of AMPKα and Bcl- 2 in cells were investigated after treated with 6 μmol/L solamargine for 24 h. RESULTS ：Compared with 020-39318678。E-mail：wujingjing6028@gzucm.edu.cn blank group ，1-12 μ mol/L solamargine for 24，48 h could significantly decrease the survival rates of cells （P＜0.05）in a concentration-dependent manner ；IC50 of them were 8.310 and 7.996 μmol/L，respectively；the rate of cell clone formation was decreased significantly after treated with 6 μmol/L solamargine for 10 days（P＜0.05）. The apoptotic rate of HepG 2 cells，mRNA expression of Bax and caspase- 3，protein expression of cleaved caspase-3（except for 8 μmol/L）as well as ratio of p-AMPKα to AMPKα（except for 8 μmol/L）were all increased significantly after treated with 6，8 μmol/L solamargine（P＜0.05）；mRNA and protein expression of Bcl- 2 were decreased significantly （P＜ 0.05）；the changes of some indexes were in a concentration-dependent manner. The compound C could inhibit protein expression of AMPKα，and reverse the inhibitory effect of solamargine on Bcl- 2 protein. CONCLUSIONS ：Solamargine can inhibit the proliferation of HepG 2 cells and induce apoptosis ，the mechanism of which may be associated with activating AMPK signaling pathway.

Regulation effects of liuwei dihuang pill, jingui shenqi pill, jiangu erxian pill containing serums on adipogenic and osteogenic differentiation-related genes expressions in the differentiation process of preadipocytes to osteoblasts / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of Chinese medical recipes for invigorating Shen on rat bone marrow mesenchymal stem cells (BMSCs)-derived preadipocytes' differentiation to osteoblasts.</p><p><b>METHODS</b>The BMSCs were cultured using whole bone marrow adherence wall method. The BMSCs were induced to preadipocytes by classic chemical method. The osteogenic differentiation process of preadipocytes was intervened by Liuwei Dihuang Pill (LDP), Jingui Shenqi Pill (JSP), or Jiangu Erxian Pill (JEP)-containing serums (with the concentRation of 10%, on behalf of tonifying Shen yin, tonifying Shen yang, and tonifying Shen essence). Reverse transcription-real time fluorescent quantitative-PCR (RT real time qPCR) was used to detect RUNX2, ALP, BGP, BMP2, BMP4, SPP1, and IGF1 mRNA expressions of osteogenic differentiation-related genes, mRNA expressions of LPL, FABP4, and PPARgamma of adipogenic differentiation-related genes on the 6th, the 12th, and the 18th day.</p><p><b>RESULTS</b>As for the osteogenic differentiation-related gene, when compared with the control group, there was no statistical difference in the gene expression level in the experimental groups on the 6th day (2.0 > Ratio > 0.5). On the 12th day, the mRNA expressions of IGF1 and Runx2 increased more significantly in the JSP group, with their relative quantification (Ratio) being 2.97 and 1.81 respectively. On the 18th day the IGF1 mRNA expression significantly increased, being the Ratio value of 3.74, 12.60, and 8.35, respectively, in the LDP group, the JSP group, and the JEP group. The SPP1 mRNA expression also significantly increased, with the Ratio value of 2.94, 3.18, and 2.62, respectively, in the LDP group, the JSP group, and the JEP group. As for adipogenic differentiation-related genes, on the 6th day, when compared with the control group, FABP4 mRNA expression significantly decreased in the LDP group and the JSP group (with the Ratio value of 0.47 and 0.40 respectively). The expression levels of other genes were all down-regulated, but not significantly. On the 12th day and 18th day, there was no statistical change in the adipogenic differentiation-related genes expressions (2.0 > Ratio > 0.5).</p><p><b>CONCLUSIONS</b>Up-regulation of osteogenic differentiation-related genes expression occurred in later time, while down-regulation of adipogenic differentiation-related genes expression occurred in earlier time after treatment by Chinese medical recipes for invigorating Shen. In general, above data indicated that tonifying Shen yang was more effective in promoting osteogenic differentiation and inhibiting adipogenic differentiation of BMSCs.</p>

Comparative study of enhancing effect on mRNA expression of hematopoietic growth factors in rat bone marrow mesenchymal stem cells by ginseng polysaccharide and ginsenoside / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study mRNA expression levels of main hematopoietic growth factors in bone marrow mesenchymal stem cells (BM-MSC), and to compare effect on mRNA expression levels treated by ginseng polysaccharide and ginsenoside.</p><p><b>METHODS</b>Relative quantification real-time polymerase chain reaction (RT-PCR) was used to observe mRNA expression levels of IL4, Csf2, Kitlg, Csf1, IL6, Lif, Csf3, IL11, Epo, and IL3, etc. in rat BM-MSC treated with ginseng polysaccharide (20 microg/mL) or ginsenoside (20 microg/mL) at 12, 24, and 36 h.</p><p><b>RESULTS</b>IL4 and Csf2 mRNA expressions were not detected. Relative expression of Kitlg, Csf1, IL6, Lif, Csf3, IL11, Epo and IL3 mRNA ranked in an attenuating order when compared with Gapdh mRNA. mRNA expression of Epo and IL3 was not significantly changed at any time point by treatment of ginseng polysaccharide or ginsenoside in rat BM-MSC (P > 0.05). mRNA expression of Csf1, IL6, Lif, Csf3 and IL11 were significantly enhanced at 12 and 36 h by treatment of ginseng polysaccharide (P < 0.05) and that of Csf1, IL6, Lif, Csf3, and Kitlg were significantly enhanced at 24 h in rat BM-MSC (P < 0.05). The enhanced mRNA expression was Csf3 at 12 h, Csf3, IL6 and Lif at 24 h, and Csf3, IL6, Lif, IL11, and Kitlg, respectively at 36 h by treatment of ginsenoside in rat BM-MSC.</p><p><b>CONCLUSIONS</b>The enhancement of ginseng polysaccharide was stronger than that of ginsenoside on mRNA expression of hematopoietic growth factors in the initial stage. As time went by, the enhancement of ginsenoside gradually increased and exceeded that of ginseng polysaccharide.</p>

The regulatory effects of polyporus polysaccharide on the nuclear factor kappa B signal pathway of bladder cancer cells stimulated by Bacillus Calmette-Guerin / 中国结合医学杂志

<p><b>OBJECTIVE</b>To detect the effects of Polyporus polysaccharide (PPS), Bacillus Calmette-Guerin (BCG), and their combination on the nuclear factor kappa B (NF-κB) signaling pathway associated-gene expression and investigate the molecular mechanisms of the toxic-reducing effect of PPS in coordination with BCG against bladder cancer.</p><p><b>METHODS</b>After T739 cells were treated with PPS, BCG and their combination, the changes in mRNA and protein expression of inhibitor of kappa B kinase beta (IKKβ), NF-κB subunit p65 (NF-κB p65), intracellular adhesion molecule 1 (ICAM1) and chemokine (C-c motif) ligand 2 (CCL2) in bladder cancer cell line T739 were determined by relative quantitative real-time PCR, Western blot, and flow cytometry (FCM). NF-κB p65 DNA-binding activity in T739 cell was detected by biotinylated probe-ELISA, and NF-κB p65 nuclear expression in T739 cell was observed by immunohistochemistry.</p><p><b>RESULTS</b>Compared with the T739 control group, the mRNA expression of IKBKB (IKKβ), Rel A (NF-κB p65), ICAM1 and CCL2 in T739 cells treated with BCG were increased obviously (Ratio>2.0), as well as the expression of IKKβ, CCL2 and ICAM1 proteins. Meanwhile, NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression in T739 cells treated with BCG were up-regulated significantly (P<0.05). Compared with the control, the increased expression in T739 cells were simultaneously down-regulated after PPS treatment, except for ICAM1 protein expression. With cells treated with a combination of BCG and PPS, the expression of genes associated with the NF-κB signaling pathway, such as IKBKB, ICAM1 and CCL2, were all down-regulated compared to the BCG group, as well as Rel A mRNA expression, NF-κB p65 DNA-binding activity and NF-κB p65 nuclear expression.</p><p><b>CONCLUSIONS</b>PPS could inhibit the over-activation of the NF-κB signaling pathway induced by BCG in bladder cancer cells and accordingly attenuate the adverse reactions to BCG therapy.</p>

Establishment of a bladder cancer cell line with toll-like receptor 2 gene knockdown and identification of its biological characteristics / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a murine bladder cancer cell line with stable silencing of toll-like receptor 2 (TLR2) expression.</p><p><b>METHODS</b>Three different recombinant plasmids pcDNA 6.2-GW/EmGFPmiR-TLR2 were constructed and transfected into T739 cells via lipofectamine. The recombinant plasmid with the strongest interference effect was selected by RT-PCR and transfected into T739 cells. The Blasticidin-resistant cell clones were screened to obtain bladder cancer cell lines with TLR2 gene knockdown, and the biological characteristics of the stable cell lines were observed.</p><p><b>RESULTS</b>Sequencing showed that the target DNA fragment was correctly inserted into the vector. The recombinant plasmid with the strongest silencing effect (pcDNA 6.2-GW/EmGFPmiR- TLR2.949) was screened, and transfection of this plasmid in T739 cells resulted in a cell line with stable TLR2 gene silencing (T739-TLR2delta), in which the expression of TLR2 mRNA and receptor were downregulated by over 95% and 90%, respectively. Compared with the negative control cells, T739-TLR2delta cell line exhibited significant different cell proliferation index with extended cell population doubling time.</p><p><b>CONCLUSION</b>The recombinant plasmid pcDNATM6.2-GW/EmGFPmiR-TLR2 can effectively suppress the expression of TLR2 gene, and the established cell line with stable TLR-2 gene knockdown allows further functional study the TLR2 gene in T739 cells.</p>

Effects of Ailing Granule on immuno-reconstruction in HIV/AIDS patients / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the possible mechanism of Ailing Granule (AG, a Chinese herbal preparation for nourishing Qi and activating blood circulation) in intervening HIV/AIDS.</p><p><b>METHODS</b>Twenty-one HIV/AIDS patients were orally administered with AG (mainly composed of Fructus Ligustri lucidi, Radix Scutellariae, Radix Astragalus and Eupolyphaga et polyphage) by 20g, twice a day for 4 months. Their symptoms and signs were scored, T cell subgroup (CD3, CD4, CD8) and interleukin-2 (IL-2) were detected with flow-cytometry, and viral load determined with RT-PCR, as well as some indexes for safety evaluation, including ALT, Cr and BW were observed before and after treatment.</p><p><b>RESULTS</b>After treatment, the symptom and sign score of patients lowered, among them the scores of fatigue, anorexia, spontaneous sweating and skin rash reduced with difference statistically (P < 0.05); the total effective rate was 61.9%; CD4 count and IL-2 level increased from 308.29 +/- 150.66/microl and 13.19 +/- 5.93 ng/L to 336.50 +/- 148.94/micro1 and 15.14 +/- 5.14 ng/L respectively( P < 0.05); while the viral load lowered but showed no significant difference. All the indexes of safety measured kept unwavering during treatment(P > 0.05).</p><p><b>CONCLUSION</b>AG could significantly alleviate the symptoms of HIV/AIDS patients, improve their immune function, inhibit HIV reproduction to a certain extent or keep it stable. No obvious toxic or adverse reaction was seen.</p>

A Report on 23 Cases of AIDS Treated with Chinese Medicine for over 10 Years / 中医杂志

Objective:To explore long-term therapeutic effect of TCM on acquired immunodeficiency syndrome(AIDS).Methods:CD_4,CD_8, CD_4/CD_8 ratio and body weight before and after treatment in the patients who had been treated decoction or proprietary medicine of TCM for over 10 years were compared.Results:CD_4 and body weight increased significantly after 30 months of treatment(P