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??OBJECTIVE To establish a method for the determination of norathyriol in rat plasma by HPLC and to study its characteristics of pharmacokinetics. METHODS Ethyl formic acid(10:1)was used to precipitate protein in the plasma samples after the addition of internal standard, and then the concentration was analyzed by HPLC. All of the separations were carried out on a Platisil ODS(4.6 mm??250 mm,5 ??m) at room temperature. The mobile phase was consisted of formic acid(40:60:0.5, pH 2.74), and was pumped at flow rate of 0.8 mL??min^-1. The UV detection wavelength was set at 312 nm. The rats were given norathriol by intragastric administration with a dosage of 400 mg??kg^-1. The concentration of norathriol in plasma at different time points was determined. RESULTS A good linear relationship was obtained in the concentration range of 0.54-162 ??g??mL^-1(r=0.998). The intra-day and inter-day RSD were less than 7.5% . The main pharmacokinetic parameters measured by Winnonlin 6.1 were showed as follows:t_max,t_1/2, ??_max and AUC_0-t were 0.5 h,(3.46??0.903) h,(26.9??3.17)??g??mL^-1,(52.4??12.0) ??g??h??mL^-1, respectively. CONCLUSION The HPLC method established is rapid and specific, and can be successfully applied in basic pharmacokinetic study in rat plasma.     

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??OBJECTIVE To explore whether knockdown serglycin's expression level in MDA-MB-231 cells by transient transfection can improve the sensitivity of breast cancer cells to the doxorubicin.METHODS At first, qRT-PCR,Western-Blot and immunofluorescence were used to examine the expression level in two different cell lines, MDA-MB-231 and MCF-7. Then serglycin's expression level in MDA-MB-231 was knocked down by using transient transfection, qRT-PCR and immunofluorescence were used to test the efficiency of this approach; and then, between the NC group and the Si-SG group, MTS was used to measure the IC₅₀ of doxorubicin and the proliferation curve under the treatment of the doxorubicin. Also the numbers of colony formation in this two groups was observed when treating with different concentration of doxorubicin. RESULTS It was found that in these two cells, the expression of serglycin in MDA-MB-231 is significantly higher than MCF-7. The efficiency of knockdown in MDA-MB-231 is above 70%. In Si-SG group, the IC₅₀ and the growth curve under treatment with doxorubicin is significantly lower than the NC group. Same RESULTS can be found in the colony formation assay, when treating with the doxorubicin, the decreasing rate of colony numbers is significantly quicker in the Si-SG group than NC group. CONCLUSION Knockdown serglycin's expression level in MDA-MB-231 cells by transient transfection can improve the sensitivity to the doxorubicin.     

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??OBJECTIVE To study the active ingredients of Flos Carthami by investigating the relationship between HPLC spectrum and antioxidant activity. METHODS The specific chromatogram of Flos Carthami processed by means of classical homothermal acceleration was assayed by HPLC. DPPH, ABTS and FRAP assays were established to determine the antioxidant activity of Flos Carthami. The spectrum-effect correlation was studied by partial least squares (PLS) method. RESULTS S₄, S₈, S₁₁, S₁₂, S₁₃, S₁₄, S₁₆ and S₁₇ were characteristic compounds in 27 matching characteristic chromatograms which had positive relationship with the ability of scavenging DPPH free radical. S₄, S₈, S₁₁, S₁₂, S₁₄, S₁₆, S₁₇ and S₁₈ were significantly positively related to the ability of scavenging ABTS free radical. S₄, S₈, S₁₁, S₁₂, S₁₃, S₁₄, S₁₆, S₁₇, S₁₈ and S₂₆ were significantly positively related to the ability of restoring Fe³⁺. Among the chromatographic peaks, S₄, S₈, S₁₁, S₁₂, S₁₄, S₁₆ and S₁₇ were positively related to the abilities of scavenging DPPH free radical and ABTS free radical and restoring Fe³⁺. S₁₄ was determined as hydroxysafflor yellow A. CONCLUSION The antioxidant material of Flos Carthami is based on the synergy of a variety of compounds. An as comprehensive as possible analysis of the compounds related to the pharmacological effect can reflect the quality of Flos Carthami. The study of spectrum-efficacy relationship is an effective way to determine the compounds related with pharmacological effect and control the quality of traditional Chinese medicinal materials.     

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??OBJECTIVE To investigate the effects of combination treatment with L-carnitine and 5-fluorouracil on the proliferation and cell apoptosis of gastric cancer MGC803 cells. METHODS MGC803 cells were divided into control group, 5-fluorouracil group and the combination of L-carnitine and 5-fluorouracil group (L-carnitine+/??5-fluorouracil group) in vitro. The inhibitory rate of cells was measured by MTT assay. The apoptosis rate and cell cycle of cells were detected by FLOW. Western blot was used to analyzed the expression of Bcl-2, Bax, adenine nucleotide translocator1(ANT1) and cleaved-PARP. RESULTS Compared with 5-fluorouracil group, the inhibition rate of MGC803 cells was increased when cells were treated with the combination of L-carnitine and 5-fluorouracil. The apoptosis rate of cells was raised and the cells were blocked at S phase. In addition, the combination group can decrease the expression of Bcl-2 and increase the expression of Bax, ANT1 and cleaved-PARP. At the same time, the apoptosis rate of cells and the cell cycle were different with the different dosage regimen when treated with the combination. Compared with the L-carnitine+5-fluorouracil group, the apoptosis rate of cells was increased to (24.17??3.12)% from (19.60??1.06)% (P<0.05). The G₀/G₁ phase proportion of cells was decreased to (62.62??1.04)% from (72.95??0.91)%,and the S phase proportion of cells was increased to (37.35??1.03)% from (27.05??0.91)% (P<0.001). CONCLUSION Treatment with L-carnitine and 5-fluorouracil could enhance the inhibitory effect of 5-fluorouracil on MGC803 cells. The possible mechanism of action is achieved by regulating the expression of Bcl-2 protein family and influencing the cell cycle.     

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??OBJECTIVE To establish an HPLC method for rapid screening and simultaneous determination of 10 preservatives in eye drops and assess their antimicrobial effectiveness. METHODS A Waters XBridge C₁₈ column (4.6 mm??150 mm,5 ??m) was used, and the mobile phase was 0.1% phosphoric acid-methanol-THF eluted in gradient mode at a flow rate of 1.0 mL??min^-1. The detection wavelength was set at 214 nm. The antimicrobial effectiveness of the preservatives was assessed according to Ch. P 2015. RESULTS The calibration curves were linear in the range of the corresponding test concentrations (r=0.999 3-1.000 0). The recoveries were between 96.1%-101.8%. One of the eye drops products did not meet the requirement of Ch. P 2015. CONCLUSION The established method is rapid and inexpensive. And it ensures excellent simplicity, sensitivity, specificity, and reproducibility and can be used for rapid screening and determination of the contents of preservatives in eye drops. The amount of preservatives should be established during the R&D period or determined according to the RESULTS of antimicrobial effectiveness test rather than using empirical values.     

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??OBJECTIVE To establish the HPLC fingerprints of Amomum villosum Lour., Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen and Amomum longiligulare T. L. Wu and find their differences. METHODS The samples were extracted with 75% ethanol aqueous and then analysis was carried out on an Agilent ZORBAX SB-Aq C₁₈ column with the mobile phase consisting of methanol (A) and 0.05% formic acid solution (B). Gradient elution (0 min, 5% A; 5 min, 5% A??15% A; 10 min, 15% A??26% A; 20 min, 26% A??40% A; 45 min, 40% A??70% A; 58 min, 70% A??100% A, 63 min, 100% A) was carried out at the flow rate of 1.0 mL??min^-1. The column temperature was maintained at 30 ??, and the detection wavelength was set at 263 nm. The software ??Similarity Evaluation System for Chromatographic Fingerprint of TCMs (Version 2012.0) ?? was employed to generate the mean chromatogras and carry out the similarity analysis of the samples. SPSS21.0 was employed to carry out the cluster analysis. RESULTS The HPLC fingerprints of the three varieties were different according to fingerprinting and cluster analysis. Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen was obvilously differernt from Amomum villosum Lour. and Amomum longiligulare T. L. Wu. There were 25 common peaks in the former HPLC fingerprint and 29 common peaks in the latter. CONCLUSION The HPLC fingerprints of three kinds of Amomum villasums were set up for the first time and they provide reference for the identification and quality control of Amomum villosum Lour., Amomum villosum Lour. var. xanthioides T. L. Wu et Senjen, and Amomum longiligulare T. L. Wu.     

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??OBJECTIVE To establish the HPLC fingerprint of Rhodiola crenulata herbs from Sichuan plateau, and compare them with commercially available samples. METHODS RP-HPLC analysis was applied using Agilent Zorbax C₁₈ chromatographic column (4.6 mm??250 mm,5 ??m). The mobile phase A was 0.1% formic acid aqueous solution and mobile phase B was 0.1% formic acid in acetonitrile. The flow rate was 1 mL??min^-1 and the column temperature was maintained at 35 ??. The detection wavelength was set at 245 nm and injection of sample was 20 ??L. The traditional Chinese medicine fingerprint chromatogram similarity evaluation system (Version 2004A), principal component analysis, and cluster analysis were used to compare 30 Rhodiola crenulata samples from various locations based on their HPLC chromatograms. RESULTS The established HPLC fingerprint of Rhodiola crenulata was able to analyze Rhodiola crenulata from different sources. CONCLUSION The method has good repeatability and stability, and can be used for the quality management standard of Rhodiola crenulata.     

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??OBJECTIVE To identify the chemical components of bile acids from bear bile powder, the intermediate of bear bile powder and Tanreqing capsules by HPLC-MS/MS. METHODS All samples were extracted with 70% methanol(V/V), and an Ultimate XB C₁₈ column (4.6 mm??250 mm, 5 ??m) was employed for separation with acetonitrile-0.1% formic acid as mobile phase in gradient elution. The MS spectrum was acquired in both positive and negative ion mode using ESI ion source. The chemical components were identified by the second mass spectrometric pyrolysis fragments, chromatographic peak retention time and fragmentation regularity summarized from the reference standards and the available literature. RESULTS A total of 33 compounds were successfully identified or tentatively predicted, and six chemical compounds including tauroursodeoxycholic acid, taurochenodeoxycholic acid, ursodeoxycholic acid, chenodeoxycholic acid, 7??-hydroxy-3-oxo-5??-cholanic acid and one unknown constituent were finally transferred to Tanreqing capsules through the intermediate of bear bile powder. Moreover, 21 new chemical compounds (major ingredients of free bile acids) were generated during the production process of the intermediate, and 19 components were also detected in Tanreqing capsules. CONCLUSION The investigation of the change of constituents in bear bile powder during Tanreqing capsules production provides a basis for the quality control and evaluation of Tanreqing capsules during production process.     

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??OBJECTIVE To prepare ion-sensitive ophthalmic in situ gel containing bendazac lysine (BDZL-ISG) and preliminarily study its rheological behavior, in vitro drug release, corneal permeation, and pharmacokinetics in rabbit aqueous humor. METHODS Single factor investigation was carried out to optimize the formulation, taking viscosity and gelling capacity as evaluation indices. Using aqueous solution or eye drops as control, the in vitro release of the formulation was evaluated by dialysis membrane method. Then, the corneal permeation experiment of the optimum formulation was carried out with Franz diffusion cell. The pharmacokinetics of BDZL-ISG in rabbit aqueous humor was preliminarily studied by microdialysis. RESULTS Compared with the control group, the optimum formulation had shear thinning behavior and significant sustained release effect. There was no significant difference in the corneal permeation between the two groups. The RESULTS of pharmacokinetic study showed that ??_max (13.25 ??g??L^-1) and AUC_0-t of BZDL-ISG were 2.38 and 2.2 times higher than those of BDZL eye drops respectively, which suggested that the ocular bioavailability of BDZL was greatly enhanced by the optimum in situ gel formulation. CONCLUSION With significant sustained release effect, the ion-sensitive ophthalmic in situ gel will become a promising alterative formulation for bendazac lysine for treatment of cataracts.     

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??OBJECTIVE To establish the quality evaluation method of the aerial roots of Ficus microcarpa L.f. based on their anti-inflammation activity. METHODS Twenty batches of samples from different producing areas were analyzed by HPLC on a Shimadzu Capcell Pak C₁₈ column(4.6 mm??250 mm,5 ??m)gradiently eluted with mobile phase of methanol and 0.1% formic acid aqueous at a flow rate of 1.0 mL??min^-1. The detection wavelength was set at 254 nm, and the column temperature was maintained at 35 ??. The chromatograms were analyzed by the software ??Similarity Evaluation System for Chromatographic Fingerprint of TCMs (Version 2012.1)?? and the common peaks were obtained. The anti-inflammatory effects of the aerial roots of Ficus microcarpa L.f. were assessed by murine model of xylene-induced ear edema. Spectrum-effect relationship was analyzed by bivariate correlation analysis using SPSS21.0. RESULTS Seventeen common peaks were identified in the HPLC fingerprints of 20 batches of aerial roots of Ficus microcarpa L.f. The anti-inflammatory effect of samples from Guangdong was better than those from Guangxi and Fujian (P<0.01). According to the result of spectrum-effect relationship analysis, six common peaks were closely related to the anti-inflammatory effects of the aerial roots of Ficus microcarpa L.f. (P<0.05). Cluster analysis was then carried out based on the six common peaks in order to divide the samples into different groups. CONCLUSION The quality evaluation method established in this research is successfully employed in the quality evaluation of the aerial root of Ficus microcarpa L.f..     

张元素药性调节观探析

将张元素药性调节观分为定向调节、定位调节、定性调节及动静调节4个方面具体分析,探索中药药性规律,为临床灵活遣方用药提供了参考.     

建立中药知识产权三维保护体系的构想

中药是我国在世界上具有潜在自主知识产权优势的少数学科之一,构建符合全球化、工业化等时代背景与自身发展要求的中药知识产权保护体系具有重要的战略意义．中药知识产权保护的内容主要有中药著作权、中药工业产权、中药商标权、中药服务标记权、中药商号名称权、中药地理标志权以及中药技术秘密等。目前中药知识产权保护存在诸如法律政策相互不协调、中药企业保护意识不强、传统技术秘密承继者市场转化意识不浓、行业保护规制缺失以及用西药的专利标准来硬套中药等问题．为此,笔者认为应该以中药理论为核心、以中药知识产权保护实务为参照、以药效为评价标准．建立中药知识产权的法律保护、行政保护和行业保护三维保护体系．以解决目前中药知识产权流失严重的局面。     

美国生物制品注册早期相关知识产权制度研究

目的 探索并分析美国生物制品注册早期相关知识产权制度.方法 通过查阅国内外文献,回顾美国《生物制品价格竞争与创新法案》颁布的背景,梳理美国利用专利舞蹈制度解决各种专利纠纷问题.结果 数据保护制度与专利并行,在12年保护期内提交生物类似药申请,美国食品药品监督管理局(FDA)还未批准的状态下,专利舞蹈制度通过生物类似药企...     

中医药知识产权保护难点及对策

中医药知识产权保护存在意识难点、申请难点和保护难点。建议在今后的工作中要加强中医药知识产权素质教育,重视培养中医药复合型人才;在知识产权保护工作中要整体谋划,提高中医药专利的技术含量。     

论蒙医药的知识产权保护

蒙医药是蒙古族人民在长期实践中总结出的宝贵经验，是我国传统知识中的一颗璀璨明珠，它具有鲜明的民族特色和神奇的功效，为中华民族特别是蒙古族人民的繁衍生息作出了巨大的贡献。首先总结了蒙医药知识的构成特点，然后分析了蒙医药知识产权保护中存在的问题，最后对蒙医药知识的保护提出了一些建议。     

从新版《药品注册管理办法》看我国药品优先审评审批制度的变化

目的 阐述我国优先审评制度的发展过程及新版《药品注册管理办法》关于加快审评审批制度,并与既往优先审评审批制度以及美国加速审评审批制度进行比较和分析,旨在为业界提供参考。方法 采用对比分析法,比较了新版《药品注册管理办法》实施前后药品加快审评审批制度的变化,并与美国加快审评审批相关制度进行对比。结果 新版《药品注册管理办法》中关于加快审评审批制度设计更加突出以临床价值为导向的鼓励创新,细化了程序和要求,使得加快审评审批制度更具可操作性。在与美国加快审评审批制度进行对比发现,新加快审评审批制度在与国际接轨的同时,还结合了我国的临床实践、患者人群范围、发病特点以及药品监管需要等,更加符合我国国情。结论 新版《药品注册管理办法》的修订将使我国药品注册加快审评审批制度更趋完善。     

论"中药知识产权保护"与中药企业的知识产权保护问题

本文简要归纳了中药产业涉及的知识产权类型，提出从产业观点出发宜对“中药知识产权”作最广义理解，并为中药企业制订知识产权保护战略提供一系列具体建议。     

浅谈中医药知识产权保护

中医药人员对中医药知识产权保护意识相对淡薄,中医药专利面临被他国抢失申请注册的现状,文章对此提出了加强知识产权知识的宣传与培训工作,把科研项目的知识产权后期管理改为前期管理等几点建议。     

针灸标准化与针灸知识产权相互关系研究

知识经济条件下,针灸标准化必然涉及知识产权。既然如此,针灸标准化与针灸知识产权二者间到底是什么关系呢?二者间的关系就是既排斥又融合的关系。如果处理不好二者间的关系,不仅会给二者各自的发展造成不利影响,还会对针灸事业的发展与繁荣造成不利影响。因此,恰当处理二者的关系,对解决诸如此类的问题具有重要现实意义。而恰当处理二者关系的前提是,必须先明了二者间的关系。本文对二者的异同做了系统比较,并对二者的相互作用做了全面剖析。     

中医药发展及其知识产权防卫策略探讨

通过对近年来中药贸易、中医药研究现状及趋势的综合分析,提出中药研究大部分是我国应当拥有的自主知识产权,强调在中医药研究开发领域知识产权保护是一项重要的工作,必须予以充分重视,并实施加强其安全防范、监管力度、专利申请与保护措施等防卫策略。