Quality comparison andstructural characteristics of material basic composition of Danshen injection from different manufacturers / 中国中药杂志

HPLC was used to analyze the chromatographic fingerprints and determine the contents of tanshinol, protocatechuic acid, protocatechuic aldehyde, caffeic acid, isoferulic acid, lithospermic acid, rosmarinic acid, salvianolic acid B, salvianolic acid A, and salvianolic acid C in Danshen injection from 10 different manufacturers. The significant differences of phenolic compounds in Danshen injection from ten manufacturers were investigated by using F test. The results showed that the similarity degree of Danshen injection from ten manufacturers was above 0.9 and there was significant difference in mass fraction of phenolic compounds between the samples from different manufacturers. The analysis of mass fraction of effective phenolic components and their structural ratios in Danshen injection from the different manufacturers showed significant differences, indicating that the Danshen injection available in market had different curative effects and with significant differences in structural ratios.

Research methods for bioequivalence and its application in Chinese materia medica preparation products / 中草药

Bioequivalence has an important role in the new drug research, development of non-patent medicine, and post-marketing evaluation of drugs and is an essential step in the standardization and modernization of Chinese materia medica (CMM). With the acceleration of modernization of CMM, CMM preparation products made by different manufacturers differ in quality, resulting in different effects. Therefore, it is very important to strengthen the application of bioequivalent study in CMM preparation products to ensure the effectiveness and safety. In this paper, the research methods of the biological equivalence were introduced systematically, and the significance of the "component structure theory" to the study on biological equivalent of the CMM preparations has been elaborated, in order to improve the evaluation system of the biological equivalent for CMM preparations.

Preparation and properties of nepeta oil-oxymatrine lipidosome pro vagina thermosensitive gel / 中草药

Objective: To prepare the nepeta oil-oxymatrine (OMT) lipidosome pro vagina thermosensitive gel, and to investigate its in vitro drug release behavior. Methods: P-407 and P-188 were used as gel matrix to prepare the gel, and gelatinization temperature was applied as a target to optimize the prescription. The OMT lipidosome was prepared based on the multiple emulsion method, and the nepeta oil-OMT lipidosome pro vagina thermosensitive gel was obtained by cold-dissolving method. The content of OMT was determined by HPLC, and in vitro release properties of nepeta oil-OMT lipidosome thermosensitive in situ gel was investigated by dialysis method. Results: After optimization, the gel prescription was finally confirmed as 18% P-407, 5% P-188, and 0.2% hydroxy-propyl methyl cellulose (HPMC). The gelatination temperature for nepeta oil-OMT lipidosome thermosensitive gel was (36.8 ± 0.2)°C, and the in vitro accumulating release ratio of sinomenine in the nepeta oil-OMT lipidosome gel system was (58.89 ± 0.34)% and (66.38 ± 0.12)% after 48 h. Conclusion: The prepared nepeta oil-OMT lipidosome thermosensitive gel has the temperature sensitivity and sustained release effect, can effectively delay the release of the drug in vagina and improve the residence time in the vagina.

ShRNA-mediated silencing of MDM2 inhibits growth of HepG2 hepatocellular carcinoma cells xenografted in nude mice / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To construct a short hairpin (sh)RNA targeting the gene encoding the MDM2 oncoprotein in order to investigate its role in human hepatocellular carcinoma (HCC) and its potential for use as a gene therapy strategy to inhibit HCC growth in vivo.</p><p><b>METHODS</b>Small interfering (si)RNAs were designed targeting the MDM2 gene (siMDM2-1 and siMDM2-2) and unrelated sequences (negative control) and cloned into the expression plasmid pGCSilencer-U6-neo-GFP. A HCC mouse model was established by subcutaneous inoculation of HepG2 cells (2 x 10(6) in 0.2 ml) into 20 nude mice. The inoculated mice were divided into four equal groups for tumor-localized injections of saline, negative control siRNA plasmid, siMDM2-1 plasmid, and siMDM2-2 plasmid. Tumor growth was observed daily (by caliper measurement) for one month, when mice were sacrificed by cervical dislocation. The tumor mass was resected for analysis of tumor inhibition rate (% = [(average tumor weight of control group - average tumor weight of treatment group) / average tumor weight of control group x 100]) and effects on MDM2 and p53 mRNA and protein expression (by reverse transcription- PCR and western blotting, both normalized to beta-actin). Significance of between-group differences was assessed by one-way ANOVA or LSD test; pairwise comparisons were made by the Chi-squared test.</p><p><b>RESULTS</b>siMDM2-1 and siMDM2-2 suppressed the xenografted tumor growth remarkably (60.6% and 54.6% inhibition rates, respectively), significantly reduced the expression ofMDM2 gene (62.8% and 61.6%) and protein (60.7% and 59.5%), and significantly increased p53 gene (47.1% and 45.6%) and protein (45.9% and 44.3%) (all, P < 0.05).</p><p><b>CONCLUSION</b>shRNA-mediated silencing of the MDM2 gene effectively inhibits HCC tumorigenesis of subcutaneously xenografted HepG2 cells in nude mice, and the mechanism may involve p53.</p>

Cbl-b and PI3K/Akt pathway are differently involved in oxygen-glucose deprivation preconditioning in PC12 cells / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Transient sublethal ischemia is known as ischemic preconditioning, which enables cells and tissues to survive subsequent prolonged lethal ischemic injury. Ischemic preconditioning exerts neuroprotection through phosphatidylinositol 3-kinase (PI3K)/Akt pathway. Cbl-b belongs to the Casitas B-lineage lymphoma (Cbl) family, and it can regulate the cell signal transduction.The roles of ubiquitin ligase Cbl-b and PI3K/Akt pathway and the relationship between them in oxygen-glucose deprivation preconditioning (OGDPC) in PC12 cells were investigated in the present study.</p><p><b>METHODS</b>Oxygen and glucose deprivation (OGD) model in PC12 cells was used in the present study. The 3-(4, 5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, nuclear staining with Hoechst 33258, and Western blotting were applied to explore the roles of Cbl-b and PI3K/Akt pathway and the relationship between them in OGDPC in PC12 cells.</p><p><b>RESULTS</b>Cell viability was significantly changed by OGD and OGDPC. OGD significantly decreased cell viability compared with the control group (P < 0.05), and preconditioning could rescue this damage was demonstrated by the increase of cell viability (P < 0.05). The expression of Cbl-b was significantly increased after OGD treatment. However, the activation of Akt and GSK3β was greatly inhibited. Preconditioning could inhibit the increase of Cbl-b caused by OGD and increase the activation of Akt and GSK3β. LY294002, a specific inhibitor of PI3K, could effectively inhibit the increase of Akt and GSK3β after preconditioning treatment. It partly inhibited the decrease of Cbl-b expression after preconditioning treatment.</p><p><b>CONCLUSION</b>Ubiquitin ligase Cbl-b and PI3K/Akt pathway are differently involved in OGDPC in PC12 cells.</p>

The study on protective effect of sphingosine-1-phosphate in cardiomyocytes / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the mechanism of protective effect of Sphingosine-1-phosphate(S1P) in cultured neonatal rat cardiomyocytes dining simulated hypoxia/reoxygenation.</p><p><b>METHODS</b>On the basis of culturing neonatal rat cardiomyocytes, the model of hypoxia-reoxygennation was built by using method of Liquid Paraffin covering, the impact of S1P on apoptosis and p-Akt and mitochondrial membrane potential were studied by using method of Propidine Iodide staining and Western blot and Bhodanmine123 staining.</p><p><b>RESULTS</b>SiP could reduce apoptosis rate (P < 0.01) and stabilize the mitochondrial membrane potential (P < 0.05) and improved the level of p-Akt1 (P < 0.01) in hypoxia/reoxygenation cardiomyocytes significantly. But wonnannin could block these effects of S1P partially.</p><p><b>CONCLUSION</b>SiP can obviously restrain apoptosis in curtured rat neonatal cardiomyocytes during simulated hypoxia/reoxygenation. Stabilization of mitochondrial membrane potential by P13K-AM signaling pathway is likely to play a role in protective action of S1P.</p>

Therapeutic effects of small interfering RNA targeting MDM2 on human osteosarcoma cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of the small interfering RNA targeting mdm2 gene on the growth of osteosarcoma cells.</p><p><b>METHODS</b>PGCsilencerTM-mdm2 siRNA was constructed and transfected into the osteosarcoma cell line U2OS cells. The inhibitory effects on mdm2 were determined by RT-PCR and Western blot analysis. The cell growth activity was determined by MTT assay, and the cell apoptosis was examined by flow cytometry. The therapeutic effects of simdm2 was assessed on the nude mouse model of transplanted tumor.</p><p><b>RESULTS</b>The simdm2 plasmid was successfully constructed. After simdm2 being transfected into the U2OS cells, the expressions of mdm2 gene and protein were significantly inhibited. The ability of cell growth activity decreased greatly and cell apoptosis occurred apparently. There was no significant difference between the negative control group and non-transfected group. The growth of xenograft tumor in simdm2 transfected nude mice was inhibited and the expressions of mdm2 gene and protein were down-regulated remarkably.</p><p><b>CONCLUSION</b>siRNA targeting mdm2 gene inhibits the mdm2 expression in osteosarcoma U2OS cells and the growth of osteosarcoma in nude mice.</p>