Objective:To preliminarily explore the synthesis of a quercetin derivative 3′,4′,5,7-four-(O-methoxy carbonyl meth-yl) quercetin and its pharmacological activities. Methods:Quercetin as the reactant and N,N-dimethyl-formamide ( DMF) as the sol-vent, the target product 3′,4′,5,7-four-(O-methoxy carbonyl methyl) quercetin was obtained by the slow addition of methyl chloroace-tate in the presence of anhydrous K2 CO3 to introduce ether bond at 3′,4′,5,7- bit. The structure was characterized by LC-MS, 1 H-NMR and element analysis. The nanoemulsion of the product was prepared using a film dispersion method, and with intraperitoneal in-jection, the effect on pituitrin-induced myocardial ischemia cardiovascular system in rats was observed. Results:3′,4′,5,7-Four-(O-methoxy carbonyl methyl) quercetin was successfully synthesized, and could be metastasized to a demethylation product containing dis-tal free carboxyl with increased polarity proved by metabolic tests in vitro. The results of electrocardiogram and animal experiments showed that the compound had improving effects on pituitrin-induced myocardial ischemia in rats. Conclusion: The nanoemulsion of 3′,4′,5,7-four-(O-methoxy carbonyl methyl) quercetin with intraperitoneal injection shows significant antagonism against pituitrin-in-duced myocardial ischemia in rats.

ObjectiveTo observe the apoptotic effect of cardamonin on K562 cells and its relationship with the expressions of PTEN, p-Akt, NF-κB and Bcl-2.Methods K562 cells were treated with cardamonin for 48 h, and the following tests were performed:(1) The cell morphology was observed by light microscopy.(2)IC50 of the K562 cells was dtermined by MTT test.(3) The apoptosis rate was detected by flow cytometry.(4) The expressions of Bcl-2 and Bax mRNA were detected a by RT-PCR.(5) The expressions of PTEN, p-Akt, NF-κB and Bcl-2 proteins were detected by Western blot.Results Obvious apoptosis was observed in the K562 cells after treated with cardamonin for 48 h.MTT assay indicated that the proliferation of K562 cells was obviously inhibited in a dose-and time-dependent manner. Comparing with the blank group, the early apoptosis rate and expression of Bax mRNA were significantly increased.At the same time, the expression of Bcl-2 mRNA was significantly decreased.All of them presented a dose-dependent manner. The expression of PTEN obviously increased with the increasing dose of cardamonin and the expressions of p-Akt, NF-κB and Bcl-2 were decreased.Conclusions Cardamonin promotes the apoptosis in K562 cells in a dose-dependent manner by increasing the expression of PTEN and decreasing the expressions of p-Akt, NF-κB, and Bcl-2.

AIM: To investigate the effect of Shenmai Injection on the expression of bFGF and PCNA of gastric cancer in mouse. METHODS: RT-PCR was used to measure the expression of bFGF and PCNA in gastric cancer cell cultured with Shenmai Injection in three concentrations.immunohistochemistry used for protein synthesis in mouse gastric cancer model. RESULTS: 140 ?L/mL of Shenmai Injection concentration inhibited 63% of bFGF and PCNA gene expression. CONCLUSION: Shenmai Injection can inhibit the express of bFGF and PCNA.

Objective To construct four expression plasmids, pcDNA3.1-GFP/HPV6bE6, pcDNA3.1-GFP/HPV6bE7, pcDNA3.1-GFP/HPV11E6, pcDNA3.1-GFP/HPV11E7 and their transfected murine cell lines. Methods The Four recombinant expression plasmids comprising HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 linked with GFP, respectively, were constructed and transfected to B16 cells by lipofectamine kit. Positive clones were selected by G418 and observed by fluorescent microscopy and identified by RT-PCR. Results The four constructed recombinant plasmids were authenticated by restriction enzyme digestion and DNA sequencing. Under the fluorescent microscope, the green fluorescence could be observed in cytoplasm and nucleus of four transfected B16 cell lines. The RNA extracted from positively transfected clones resistant to G418 were analyzed by RT-PCR, which demonstrated the presence of four expected fragments. Conclusions The transfected murine cell lines B16 can express HPV6bE6,HPV6bE7,HPV11E6 and HPV11E7 gene. These transfected cell lines can be further transplanted to mice in order to investigate the biological properties and immunological mechanisms of these genes in vivo.

Objective: To investigate the effect of Shenmai Injection (Radix Ginseng Rubra, Radix Ophiopogonis) on angiogenesis and the expression of proliferating cell nuclear antigen (PCNA) in tumor tissue, and the mechanism of it in the treatment of tumor. Methods: The mouse tumor model was used to investigate the effect of Shenmai Injection on tumor growth on the whole. The expression of von Willebrand factor (vWF) and PCNA in tumor tissue was studied by means of immunohistochemistry. Results: Shenmai inhibited the tumor growth and reduced microvessel density and the expression of PCNA in tumor tissue. Conclusion:Antiangiogenesis is one of the mechanisms of Shenmai Injection in treatment of tumor.

OBJECTIVETo obtain transgenic Pinellia ternata plants resistant to fungus by transfer Chitinase and beta-1,3-Glucanase gene from Trichoderma harzianum.

METHODUsing hygromycin phosphotransferase as the selection marker, the Chitinase gene (ech42), beta-1,3-Glucanase gene (gluc78) and both gene pCAMBIA(ech42 + gluc78) driven by CaMV35S promoter were transferred into P. ternata callus via Agrobacterium-mediated transformation.

RESULTPCR results confirmed that the regenerants were identified to be transgenic lines and the RT-PCR results confirmed that foreign genes construction were transfer to mRNA. Two foreign genes were inherited stably to T5 generation according to PCR results of the lines.

CONCLUSIONThe results showed that chitinase gene ech42 and beta-1, 3-glucanase gene gluc78 respectively or together introducing and co-integrating into P. ternata

Agrobacterium tumefaciens ; genetics ; metabolism ; Chitinases ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Gene Expression Regulation, Plant ; Gene Transfer Techniques ; Genetic Vectors ; genetics ; metabolism ; Glucan 1,3-beta-Glucosidase ; genetics ; metabolism ; Pinellia ; genetics ; metabolism ; Transformation, Genetic ; Trichoderma ; enzymology

Background The metabolites and metabolic pathways of hand-arm vibration syndrome have not yet been elucidated. Objective To investigate the effect of local vibration on endogenous metabolites in rat serum by metabolomic analysis, to preliminarily explore the potential metabolic pathway of endogenous metabolites, so as to provide evidence for further research on the mechanism of hand-arm vibration syndrome. Methods Thirty-two SPF male SD rats, (211.3±11.1) g, 7−8 weeks of age, were selected and randomly divided into three groups: control group (14 rats, without vibration), 7 d vibration group (9 rats, continuously vibration for 7 d), and 14 d vibration group (9 rats, continuous vibration for 14 d). The vibration rats were vibrated every day for 4 h, the frequency weighted acceleration was 4.9 m·s⁻², the vibration frequency was 125 Hz, and the vibration direction was one-way vertical vibration. The control group had the same conditions except not contacting vibration. After the vibration exposure, the blood samples taken from the abnormal aorta of rats were collected, and the changes of rat serum metabolome were analyzed by ultra-performance liquid chromatography-tandem time-of-flight mass spectrometry. Principal components analysis (PCA) was used to explore changes in rat serum metabolic profile, and orthogonal partial least squares-discriminant analysis (OPLS-DA) was used to screen out differential metabolites. Combined with online databases, a metabolic pathway enrichment analysis of differential metabolites was performed. Results The PCA analysis showed that compared with the control group, the rat serum metabolic profiles in the 7 d group and the 14 d group were clearly differentiated, and the rat serum metabolic profiles in the 7 d group and the 14 d group partially overlapped. The OPLS-DA analysis showed significant differences between groups. The main parameters were: model interpretation rate R²Y=0.914, model predictive ability Q²=0.58. The OPLS-DA analysis screened out 26 and 119 differential metabolites from the 7 d group and the 14 d group respectively, and there were 24 common differential metabolites between the 7 d group and the 14 d group. The metabolomic pathway analysis showed that local vibration-induced changes in rat serum metabolism were mainly related to arachidonic acid metabolism in the 14 d group, among which the metabolites with significant effects were arachidonic acid, prostaglandin E₂, and prostaglandin D₂. Conclusion Local vibration could affect the normal metabolism in rats, and the metabolic pathway with significant influence is arachidonic acid metabolism after a 14 d exposure and the involved metabolites are arachidonic acid, prostaglandin E₂, and prostaglandin D₂.