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Objective:To investigate the effects of total flavonoids from Cycas Revolute on expression of vascular endothelial growth factor (VEGF),basic fibroblast growth factor (bFGF),Hypoxia inducible factor-1α (HIF-1α) and nuclear factor-κB(NF-κB) in model mice of Lewis lung cancer.Methods: The expressions of VEGF,bFGF,HIF-1α and NF-κB in tumor tissues were detected by immunohistochemistry and Western blot.The expression of VEGF,VEGF and NF-κB in tumor tissues were detected by fluorescent quantitative PCR.BFGF,HIF-1α and NF-κB mRNA were detected by immunohistochemistry.Results: The results of immunohis-tochemistry,Western blot and Real-time PCR showed that the results were basically the same,compared with model group,the expression of VEGF,bFGF,HIF-1α,NF-κB mRNA and the expression of VEGF,bFGF,HIF-1α and NF-κB were decreased,the difference was highly significant (P<0.05).Conclusion: The mechanism of total flavonoids from Cycas Revolute in the treatment of lung cancer may be through inhibition of the expression of VEGF,bFGF,HIF-1α,NF-κB in invasion and metastasis,and further inhibit the expression of VEGF,bFGF,HIF-1α and NF-κB in invasion and metastasis-related proteins,thus play a role of anti-lung cancer invasion and metastasis.
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OBJECTIVE:To optimize the ultrasonic extraction and purification technology of total saponins from Marsdenia te-nacissima,and to prepare purified total saponins from M. tenacissima. METHODS:Using the extraction rate of total saponins from M. tenacissima as index,ethanol volume fraction,solvent-solid ratio and extraction temperature as factors,ultrasonic extraction technology of total saponins from M. tenacissima was optimized by single factor test and Box-Behnken response surface design. Us-ing the content of total saponins in elution,the effects of ethanol elution volume fraction on purification technology of total sapo-nins by D101 type macroporous adsorption resin was investigated. RESULTS:The optimal ultrasonic extraction technology was as follows as ethanol volume fraction of 85%,solvent-solid ratio of 18.5∶1 (ml/g),extraction temperature of 80 ℃. In verification test,the average extraction rate of total saponins from M. tenacissima was 4.80%(RSD=1.06%,n=3);it was close to predicted value 4.89%,and the deviation was 1.84%. The optimal purification technology was as follows as 45% ethanol as eluant,90%ethanol for purification. Verification test showed that the average content of total saponins from M. tenacissima was 62.45%(RSD=0.88%,n=3). CONCLUSIONS:The optimized technology can be used for the extraction and purification of total saponins from M. tenacissima. The technology is stable and reliable.
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Gynostemma pentaphyllum belonged to the Cucurbitaceae family.It was also called theQi-Ye-Dan,licorice stem,Gong-Luo Guo-Di,and etc.It contained saponin,flavonoids,polysaccharides,terpenoids and other chemical compositions.Its pharmacological effects included cholesterol-lowering activity,antitumor activity,hypoglycemic activity,anti-aging and boost immunity.This paper reviewed the research progress on chemical constituents,pharmacological effects and in vivometabolism of gynostemma pentaphyllum,in order to provide a theoretical basis for its further research and development.
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Objective To study the Dai medicine Yapa Tang inhibition on mice Lewis lung carcinoma growth and metastasis,and to explore the impact of the drug on mice immune function. Methods Subcutaneous injection LLC(Lewis lung carcinoma cells)in C57BL/6 mice’s right armpit was to establish rat models. 60 successful tumor-bearing mice were selected and thenafter inoculating LLC were randomly divided into 6 groups, namely the control group,Yapa Tang low-dose group,Yapa Tang mid-dose group, Yapa Tang high-dose group,cyclophosphamide group and Yapa Tang mid-dose combining cyclophosphamide group.Then,after inoculated 3 days,each group was administrated with medicines.The first four groups wereintragastricly administrated with 0.9% saline, low-dose, mid-dose and high-dose Yapa Tang with0.4ml respectively;the cyclophosphamide group was administrated with cyclophosphamide,and Yapa Tang mid-dose combining cyclophosphamide group was administrated with middle dose Yapa Tang and cyclophosphamide. After 21days, blood was taken from eyeballs. Tumor tissue, spleen, thymus were gotten and weighted to calculate inhibitory rate,spleen and thymus index, enzyme-linked immunosorbent assay (Elisa) was adopted to detect Interleukin-2(IL-2), Interleukin-10(IL-10)and their contents. Results The tumor weight of Yapa Tang high-dose group, cyclophosphamide group and Yapa Tang mid-dose combining cyclophosphamide group was 3.46±0.39, 2.39±1.04 and 2.30±0.76 respectively,all lower than the control group, which was 5.21±0.50, showing significant difference(P<0.05), with the inhibition rate were33.69%, 54.12%, 56.00%. The thymus and spleen index of Yapa Tang low-dose group, middle-dose group and high-dose group was2.16±0.69, 2.24± 0.76, 2.23 ± 0.63, 16.82 ± 3.14, 15.82 ± 1.72, 17.08 ± 3.65, significantly higher than that of the control group,which was1.94±0.6, 15.17±3.53. The IL-2, IL-10 level of control group were(883.54±181.49)ng/L, (1 106.86±343.79)ng/L. Yapa Tanggroupsshowed an increase in IL-2(1 732.29±100.52)ng/L, (1 813.33± 168.32)ng/L, (2 275.63 ± 394.76)ng/L and the decrease in level of IL-10, respectively were(834.02 ± 271.97)ng/L, (636.83±270.56)ng/L, (682.08±147.85)ng/L, with significant difference. Conclusion Yapa Tang can inhibit lewis lung cancer grow and reduce the number of lung metastases, regulate immune cytokines IL-2 and IL-10, and promote the immunity of tumor-bearing mice.
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Objective To study the anti-tumor effect of tenacissoside H on Lewis lung cancer mice, and explore its impacts on immune function of tumor-bearing mice. Methods Mice was injected Lewis lung cancer cells subcutaneously to the right axilla. Thirty successful tumor-bearing mice were randomly divided into model group, the cyclophosphamide (CTX) group and tenacissoside H group, 10 mice in each group. Three days after inoculation, mice were intraperitoneally injected by normal saline, CTX and tenacissoside H respectively every two days, 0.2 mL in each mouse. On the 21st day, the eyeballs were extracted and blood was drawn, tumor tissue, spleen and thymus were taken and weighted to calculate tumor inhibition rate, spleen index and thymus index, and the contents of IL-2 and IL-10 were detected by ELISA. Results The tumor weights of CTX group and tenacissoside H group were lower than that of the model group with significant difference (P<0.05), and the tumor inhibition rates were 54.12%, 25.68%. The thymus index and spleen index of tenacissoside H group increased, but that of CTX group decreased significantly. Compared with the model group, the IL-2 level of tenacissoside H group was significantly increased, while the IL-10 level decreased. Conclusion Tenacissoside H can inhibit growth and metastasis of Lewis lung cancer, regulate the expression of IL-2 and IL-10, and improve the immune function of tumor-bearing mice.
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<p><b>OBJECTIVE</b>To establish a culture system for Psammosilene tunicoides hairy roots, and provide technological aid for the large-scale production of P. tunicoides material.</p><p><b>METHOD</b>The young leaves and stem segments of sterile plantlets were infected with ACCC10060 strain, and subsequently a culture system suitable for hairy roots growth was further established.</p><p><b>RESULT</b>When explants were co-cultured with ACCC10060 (A600 0.8) on B5 media containing 20 mg x L(-1) Acetosyringo (AS) for 48 h, the hairy roots could be successfully induced, and it could achieve a higher induction rate using young leaves as explants than that of stem segments. The transfected hairy roots possessed the ability of kanamycin resistance and growth on hormone-free media, and synthesis of opines. All above results demonstrated that the present hairy roots originated in the infection of P. tunicoides tissues by ACCC10060 strains. After 35 d culture in liquid hormone-free MS (1/2 strength), the biomass of hairy roots increased 14.11 times (fresh weight) and 8. 39 times (dry weight), respectively, and the content of total saponins in hairy roots reached to 0.857% (DW), by contrast, it's only 0.388% and 0.217% in callus and seedlings respectively.</p><p><b>CONCLUSION</b>Establishment of hairy roots culture of P. tunicoides provided a foundation for industrial production of active components from P. tunicoides culture.</p>
Subject(s)
Biomass , Caryophyllaceae , Microbiology , Culture Techniques , Plant Roots , Microbiology , Rhizobium , Physiology , Saponins摘要
OBJECTIVE: To establish the quality standard of Yuquan capsules. METHODS: The composition of Yuquan capsules, such as Puerariae lobata, Panax ginseng, Glycyrrhiza uralensis, Astragalus membranaceus, Schisandrae chinensis, were identified by TLC. The content of puerain was determined by HPLC. RESULTS: The identified components of Yuquan capsules were P. lobata, P. ginseng, G. uralensis, A. membranaceus, S. chinensis. The linear range of puerain were 0.127 5~1.02 ?g(r=0.999 9) with an average recovery of 100.29%(RSD=1.68%). CONCLUSION: Established standard can be used for the quality control of Yuquan capsules.
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Objective To establish an ICP-AES method for determination of the content of Zn in Shenningsan Capsules. Methods The sample was digested with HNO3-HClO4, and determined by the ICP-AES method. The determined wavelength was at 213.8 nm, the electric current of the light was set at 10.0 mA, the slit-width was 0.5 nm, the flow rate of acetylerce was 2.0 L/min and the flow rate of air was 9.4 L/min, the hight of burning was 7.5 mm. Results The ion concentration of Zn had the good linear range of 0.2~1.6 ?g/mL, Y=0.054X+0.001 1 (r2=0.999 3). The average recovery of the content of Zn in Shenningsan Capsules was 99.56% with RSD of 2.00%. The content of Zn in Shenningsan Capsules was 68.6 ?g/g. Conclusion The method is simple and reliable. It can be used effectively for the quality control of Shenningsan Capsules.
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Objective To search for the optimal processing procedure and physico-chemical properties of Endothelium corneum amylase. Methods After dried under 50 ℃ ,amylase was extracted from Endothelium corneum with phosphate-buffered saline. The amylase activity was examined at different temperature,pH and different metal ions,and amylase isozyme was discerned on PAGE. Results The optimal pH,optimal temperature,and optimal substrate concentration for amylase activity were 8.67,50 ℃ and 1.8 g/mL,respectively. Under the experimental conditions,the amylase Km was 0.92 (starch was used as substrate of amylase). Four metal ions tested,including Ca2+,Fe2+,Zn2+,Mn2+,could inhibit the amylase activity to some extent. Furthermore,it showed one amylase isozyme band using PAGE,suggesting only one amylase isozyme existing in the Endothelium corneum tentatively. Conclusion The effects of metal ions should be considered in the process of Endothelium corneum amylase.
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Objective To establish a method for the determination of AstragalosideⅣin Ejiao Buxue Koufuye.Methods HPLC-ELSD was used to determine directly AstragalosideⅣin Ejiao Buxue Koufuye on Betasil C_(18) column,using CH_3OH-H_2O(74:26)as a mobile phase.The flow rate was 1.0mL/min.The temperature of drift tube for ELSD was 90℃and the flow rate of N_2 was 2.0L/min.Results The linear range of AstragalosideⅣwas 0.43~2.15?g,Y=1.53637X+0.92158,r=0.9999(n=6),the average recovery of the added sample was 98.5% and RSD=0.98%.Conclusion The method is accurate and reliable with a good reproducibility.It can be used for the quality control of Ejiao Buxue Koufuye.
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OBJECTIVE: To compare the content of taxol in stalk,leaf and callus of Taxus cuspidata.METHODS:Dichloromethane and water extract were extracted form T.cuspidata using methanol-chloroform(1 ∶ 1).HPLC method was used to determine the content of taxol.RESULTS: The content of taxol in stalk,leaf and callus of T.cuspidata were 0.018 30%,0.001 28%,0.000 66%,respectively.The content of taxol in the callus of T.cuspidata is higher than other two parts of it.CONCLUSION:This study is significance for the development and utilization of T.caspidate and its protection.