肠道菌群对中药糖苷类成分脱糖基代谢的研究进展

中药糖苷类成分是中药中一类重要的活性物质,其药理活性、药代动力学特征及体内存在是目前研究的热点。糖苷类成分在生物体内代谢转化途径主要是由肠道菌群介导的脱糖基代谢,生成苷元后更易吸收入血并发挥药效。该文基于肠道菌群在中药糖苷类成分体内代谢及药效发挥中的重要作用,综述了肠道菌群中代谢中药糖苷类成分的主要糖苷酶,产生糖苷酶的主要菌属,以及代表性糖苷类成分的脱糖基代谢途径。并针对肠道菌群对中药糖苷类成分代谢研究过程中存在的问题进行了初步探讨。

药物与肠道菌群的相互作用

肠道菌群是一个复杂的微生态系统,在维持宿主的内环境稳态、保障宿主健康方面发挥了十分重要的作用;同时,肠道菌群与疾病的发生、发展息息相关,也与药物的治疗作用、毒性反应密切相关。药物进入体内后,在肠道菌群的作用下发生脱糖基等代谢反应,使药物成分的活性或毒性发生改变;同时,在药物作用下,肠道菌群的种类和数量发生了改变,药物间接发挥治疗作用或产生毒性反应。该文从药物与肠道菌群相互作用的两个方面"肠道菌群对药物的处置"和"药物对肠道菌群的作用"进行了系统的综述,揭示药物在肠道菌群介导下发挥治疗作用或产生毒性的效应及机制。

Selection of Effective Bcl-2 Antisense Oligodeoxynucleotides with Computer Software and Experimental Assay

Objective： To explore and investigate the selection of effective antisense oligodeoxynuleotides with the help of computer and RNAstructure folding software. Methods： Bcl-2 gene was used as the target gene and five antisense oligodeoxynuleotides were designed to be bound to Bcl-2 mRNA optimal secondary structure regions that were predicted free from intramolecular fold or instability of free energy. The five antisense oligodeoxynucleotides were studied with experimental assay of leukemia cells, including cell grow assay with tropan blue exclusion, expression of Bcl-2 protein detected with immunochemistry and flowcytometry, Bcl-2 mRNA content detected with RT-PCR technique, as well as apoptosis observed and determined with morphonological method, electrophoresis and flowcytometry. Results： The results showed that two of the five antisense oligodeoxynucleotides were effective antisense oligodeoxynucleotides, which were able to inhibit cell growth in leukemia, to decrease the level of Bcl-2 mRNA and protein, to induce apoptosis of leukemia cells significantly. Conclusion： The computational prediction of antisense efficacy is faster than other methods and more efficient, which can potentially speed the development of sequences for both research and clinical applications.

Establishment of the Agrobacterium-mediated Genetic Transformation System of Ginkgo biloba and the Construction of the Expression Vector of Gb-DXR

[Objective] The research aimed to provide reference for increasing the genetic transformation efficiency of Ginkgo biloba mediated by Agrobacterium.[Method] Taking the mature embryos of Ginkgo biloba seeds as explants,after 48 hours＇ pre-cultivation on MS medium in the absence of phytohormone,GUS gene was transmitted into embryos of Ginkgo biloba mediated by three kinds of Agrobacterium.Transient expression of GUS gene activity was observed through histochemical staining,and the influencing factors of the expression of GUS gene were analyzed.And the expression vector of 1-deoxy-D-xylulose-5-phosphate reductoisomerase in the biosynthesis approach of biobalide precursor of Ginkgo biloba was constructed.[Result] A more suitable genetic transformation scheme was obtained as follows：taking embryos of Ginkgo biloba as explants,using EHA105 Agrobacterium with pCAMBIA1304＋ for infection,co-culture for 3 days and GUS staining.The results showed that transient expression rate of GUS after transformation was higher.[Conclusion] The research provide a more effective method for further study on the transgene of Ginkgo biloba.

Homologous recombination in DNA repair and DNA damage tolerance

Homologous recombination （HR） comprises a series of interrelated pathways that function in the repair of DNA double-stranded breaks （DSBs） and interstrand crosslinks （ICLs）. In addition, recombination provides critical support for DNA replication in the recovery of stalled or broken replication forks, contributing to tolerance of DNA damage. A central core of proteins, most critically the RecA homolog Rad51, catalyzes the key reactions that typify HR： homology search and DNA strand invasion. The diverse functions of recombination are reflected in the need for context-specific factors that perform supplemental functions in conjunction with the core proteins. The inability to properly repair complex DNA damage and resolve DNA replication stress leads to genomic instability and contributes to cancer etiology. Mutations in the BRCA2 recombination gene cause predisposition to breast and ovarian cancer as well as Fanconi anemia, a cancer predisposition syndrome characterized by a defect in the repair of DNA interstrand crosslinks. The cellular functions of recombination are also germane to DNA-based treatment modalities of cancer, which target replicating cells by the direct or indirect induction of DNA lesions that are substrates for recombination pathways. This review focuses on mechanistic aspects of HR relating to DSB and ICL repair as well as replication fork support.

Binding and Transformation of Extracellular DNA in Soil

DNA is the genetic material of various organisms. Extracellular DNA adsorbed or bound on surface-active particles in soils has been shown to persist for long periods against nucleases degradation and still retain the ability to transform competent cells. This paper reviews some recent advances on the binding and transformation of extracellular DNA in soils, which is fundamental to understanding the nature of the soil, regulating biodiversity, and assessing the risk of releasing genetically engineered microo…

中药皂苷类化合物生物转化研究进展

皂苷广泛存在于药用植物中,按母核结构可分为三萜和甾体皂苷,具有多种药理活性。众多研究发现皂苷中糖基部分的组成与其药理活性密切相关,改变皂苷分子中的糖基数量和结构能改变皂苷的药理活性及其生物利用度,为开发药物提供更多的化合物资源。相较于传统的化学方法,皂苷生物转化修饰糖基部分具有选择性高、成本低、污染小等优点,被广泛用于次级皂苷与皂苷元的制备。通过对皂苷脱糖基转化生成较高生物活性的次级皂苷与苷元的研究进展进行综述,为皂苷类化合物生物转化研究提供思路和方法。

PROTECTION AGAINST LEPTOSPIROSIS BY IMMU NIZATION WITH PLASMID DNA ENCODING 33 kDa ENDOFLAGELLIN OF L.INTERROGANS SEROVAR LAI

To evaluate how the efficacy of DNA inocutation affects the ability to raise protective immunity against Leptospira. [WT5”BX]Methods.[WT5”BZ] A pair of oligonucleotide primers were designed to amplify the endoflagellar gene of L. interrogans sensu stricto serovar lai. An approximately 840bp fragment was generated with PCR and inserted into VR1012, a plasmid DNA expression vector, after the fragment and VR1012 were digested respectively with EcoRV and Sal I. A recombinant plasmid designated as VR1012+flaB2 was obtained. The vector, VR1012 consits of a pUC18 backbone with the cytomegalovirus(CMV) IE1 enhancer, promoter, and intron A, transcription regulatory elements and the BGH polyadenylation sequences driving the expressing of leptospiral endoflagellar gene of L. interrogans sensu stricto serovar lai. Plasmid encoding leptospiral endoflagellin gene was injected into quadriceps of NZW rabbits. [WT5”BX]Results.[WT5”BZ]This resulted in the generation of specific leptospiral antibody with high ELISA titer (1:32768) in the rabbits. Immuno/protection was performed in guinea pigs without adjuvant. The group“VR1012+flaB2” showed higher survival rate(90%,9/10 animals),compared with the group “VR1012 lack flaB2” and the group “normal saline”. [WT5”BX]Conclusion.[WT5”BZ]The technique of DNA vaccine has potential advantages over certain other vaccine preparation technologies. However whether DNA vaccine will be useful for vaccine development remains to be tested.

Determination of 2-Amino-6-Cyclopropylamino-9-(2,3-Dideoxy-β-D-glyceropent-2-enofuranosyl)purine in Rat Plasma,Urine and Liver Homogenates by High Performance Liquid Chromatography

Aim To develop a simple and specific high-performance liquid chromatographic(HPLC) method, suitable for the pharmacokinetic studies in vivo, to determine the concentrations of2-amino-6-cyclopropylamino-9-(2,3-dideoxy-β-D-glyceropent-2-enofuranosyl)purine (Cyclo-D4G, IMGprodrug) in rat plasma, urine and liver homogenates. Methods Chromatography was performed with C-18Hypersil ODS column and a mobile phase of 7% (v/v) acetonitrile in phosphate buffer, pH 7.40, withUV detection at 283 nm. Results The average extraction recovery of Cyclo-D4G in rat plasma and urinewas 100.1% over its linear range of 0.5 - 80 μg·mL^(-1). The accuracy of the assay was 99.4% .The intra-and inter-day RSDs were less than 9.0% . Conclusion The analytical method was found to beapplicable, reliable and suitable for pharmacokinetic studies.

Design and activity evaluation of deoxyribozymes specifically targeting hepatitis C virus RNA

Objective: To explore the cleaving and inhibitory activity of hepatitis C virus ( HCV) -specific deoxyri-bozymes (DRz) at both molecular and transgeneic cellular levels. Methods: According to the secondary structure of HCV 5'-noncoding region (5'-NCR) and the sites characterized with 5'…Y ↓ R…3'(Y = A/G,R = U/C) , HCV-spe-cific naive deoxyribozymes were designed and named DRz-232, DRz-127, DRz-84, DRz1, and the phosphorothioate deoxyribozymes (PSDRz) and mutated phosphorothioate deoxyribozymes (MPSDRz) were also designed. HCV RNA 5'-NCR was transcribed in vitro from linearized plasmid pHCV-neo and radiolabelled at its 5'-end. DRz, PSDRz or MPSDRz was respectively mixed with the substrate RNA and incubated under appropriate conditions, the cleaved products were displayed by 8% denaturated polyacrylamide gel electrophoresis (PAGE) and autoradiography, and the optical density of each band was measured to calculate cleavage rates. After that, every kind of DRz was added respectively to the cultured transgeneic HepG2 cells containing luciferase gene controlled by HCV 5'-NCR. The cells were lysed at intended time points and the activity of luciferase was measured with chemiluminescence method for calculating inhibition rates. Results: After incubated for 90 min in vitro, the cleavage rates of DRz-127, PSDRz-127, DRz1 and PS-DRz1 reached 32.6% , 30. 8% , 24. 3% and 21. 5% , respectively. No cleavage product was observed in any MPSDRz. DRz-127, PSDRz-127, DRz1 and PSDRzl had an inhibitory rate of 53. 2% , 50. 6% , 44. 7% and 43. 3% respectively in transgeneic HepG2 cells in the first 24 h when the final dose of the DRz was 0. 5μmol/L, higher than that of the corresponding MPSDRz. There was no significant difference between the inhibitory effect of each DRz and its PSDRz in HepG2 cells, but the inhibitory rate of DRz decreased more rapidly than that of the latter with the elapse of time. The results from transfection groups were significantly better than those of non-transfection groups. Conclusion: Rationally-designed HCV-specific deoxyribozymes are able to cleave target RNA at molecular level in vitro, and efficiently inhibit the expression of luciferase gene controlled by HCV 5'-NCR in transgeneic cells. Appropriate PSDRz may be more stable, and thus more suitable than the naive DRz in the application to cells. Introduction of the deoxyribozymes with transfection is more efficient than with direct delivering ways.

STUDY ON GMA-DNA ADDUCTS

Objective. DNA modification fixed as mutations in the cells may be an essential factor in the initiation step of chemical carcinogenesis. In order to explore the mechanism of gene mutation and cell transformation induced by glycidyl methacrylate (GMA), the current test studied the characteristics of GMA DNA adducts formation in vitro. Methods. In vitro test, dAMP, dCMP, dGMP, dTMP and calf thymus DNA were allowed to react with GMA (Glycidyl Methacrylate). After the reaction, the mixtures were detected by UV and subjected to reversed phase HPLC on ultrasphere ODS reversed phase column, the reaction products were eluted with a linear gradients of methanol (solvent A) and 10mmol/L ammonium formate, pH5 0 (solvent B). The synthesized adducts were then characterized by UV spectroscopy in acid (pH1 0), neutral (pH7 2), alkaline (pH11 0) and by mass spectroscopy. Results. The results showed that GMA could bind with dAMP, dCMP, dGMP and calf thymus DNA by covalent bond, and the binding sites were specific (N 6 of adenine, N 3 of cytosine). Meanwhile, a main GMA DNA adduct in the reaction of GMA with calf thymus DNA was confirmed as N 3 methacrylate 2 hydroxypropy1 dCMP. Conclusions. GMA can react with DNA and /or deoxynucleotide monophosphate and generate some adducts such as N 6 methacrylate 2 hydroxypropyl dAMP and N 3 methacrylate 2 hydroxypropyl dCMP, ets. Formation of GMA DNA adducts is an important molecular event in gene mutation and cell transformation induced by GMA.

EVOLUTIONARY RELATIONSHIPS AMONG SEVENTEEN HUMAN PAPILLOMAVIRUS GENOTYPES

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STUDIES ON THE SYNTHESES AND PROPERTIES OFSNAKE-CAGE TYPE CHELATE RESINS I System of Carboxymethyl-chitosan/B-62/Triethylenetetramine

Using carboxymethyl chitosan （CM-CTS） as snake resin, B-62 resin crosslinked by,triethylenetetramine （TETA） as cage resin, a series of novel snake-cage type resin weresynthesized. Such factors as the best synthetic conditions, the swelling and regenerationproperties, and the sorption capacities of the above mentioned resins for metal ions wereinvestigated. The experimental results show these resins have good swelling propertiesand mechanical stability and do not run off in organic and inorganic solvents. Thesorption capacities of them for Cu2+, Ni2+,Zn2+, and Pb2+ were 0.89, 0.54, 0.32, and0.22mmol/g, respectively.

Decreased mitochondrial deoxyribonucleic acid and increased oxidative damage in chronic hepatitis C

AIM： To determine whether alteration of the mito- chondria DNA （mtDNA） copy number and its oxidative damage index （mtDNA△CT） can be detected by analysis of peripheral blood cells in hepatitis C virus （HCV）- infected patients. METHODS： This study enrolled two groups of pa- tients aged 40-60 years： a control group and an HCV- infected group in Department of Gastroenterology and Hepatology in Changhua Christian Hospital. Patients with co-infection with hepatitis B virus or human im- munodeficiency virus, autoimmune disease, malignant neoplasia, pregnancy, thyroid disease, or alcohol con- sumption 〉 40 g/d were excluded. HCV-infected pa- tients who met the following criteria were included： （1） positive HCV antibodies for 〉 6 mo; （2） alanine aminotransferase （ALT） levels more than twice the upper lim- it of normal on at least two occasions during the past 6 mo; and （3） histological fibrosis stage higher than F1. The mtDNA copy number and oxidative damage index of HCV mtDNA （mtDNA△CT） were measured in periph- eral blood leukocytes. The association between mtDNA copy number and mtDNA△CT was further analyzed using clinical data. RESULTS： Forty-seven normal controls （male/female： 26/21, mean age 50.51 ± 6.15 years） and 132 HCV- infected patients （male/female： 76/61, mean age 51.65 ± 5.50 years） were included in the study. The geno- types of HCV-infected patients include type 1a （n = 3）, type 1b （n = 83）, type 2a （n = 32）, and type 2b （n = 14）. Liver fibrosis stages were distributed as follows： F1/F2/F3/F4 = 1/61/45/25 and activity scores were A0/ A1/A2/A3 = 7/45/55/25. There were no age or gender differences between the two groups. HCV-infected pa- tients had higher hepatitis activity （aspartate transami- nase levels 108.77 ± 60.73 vs 23.19 ± 5.47, P 〈 0.01; ALT levels 168.69 ± 93.12 vs 23.15 ± 9.45, P 〈 0.01） and lower platelet count （170.40±58.00 vs 251.24 ± 63.42, P 〈 0.01） than controls. The mtDNA copy num- ber was lower in HCV-infected patients than in controls （173.49 vs 247.93, P 〈 0.05）. The mtDNA△CT was higher in HCV-infected patients than in controls （2.92 vs 0.64, P 〈 0.05）. To clarify the clinical significance of these results in HCV-infected patients, their association with different clinical parameters among HCV-infected pa- tients was analyzed. A negative association was found between mtDNA copy number and elevated aspartate transaminase levels （r = -0.17, P 〈 0.05）. Changes in mtDNA copy number were not associated with HCV RNA levels, HCV genotypes, liver fibrosis severity, or inflammatory activity in the liver biopsy specimen. How- ever, a correlation was observed between mtDNA△CT and platelet count （r = -0.22, P 〈 0.01）, HCV RNA level （r = 0.36, P 〈 0.01）, and hepatitis activity （r = 0.20, P = 0.02）. However, no difference in the change in mtDNA△CT was observed between different fibrosis stages or HCV CONCLUSION： Oxidative stress and mtDNA dam- age are detectable in patient＇s peripheral leukocytes. Increased leukocyte mtDNA△CT correlates with higher HCV viremia, increased hepatitis activity, and lower platelet count.

Molecular modeling study of the effect of base methylation on internal interactions and motions in DNA and implication to B-Z conformation change

Methylation in the bases of DNA is known to induce B-Z conformation change. In this work, molecular mechanics and normal mode analysis are used to probe how certain methylation affects the internal interactions and thermodynamic motions in the DNA double helixes in both B and Z conformations, and its implication to B-Z conformation change. By molecular modeling with Insight II, two cases involving cytosine C5 and guanine C8 methylation on both B and Z-form DNA duplex d(CGCGCG)2 are studied in comparison with the corresponding unmethylated duplexes. The internal interaction energies computed based on a molecular mechanics force field and the entropies due to internal motions computed according to a normal mode analysis are in fare agreement with respective observed thermodynamic quantities. The analysis on the computed individual energy terms suggests that the observed B-Z conformation change induced by methylation is primarily driven by enthalpic factors. A combination of changes in Van der Waals interaction, electrostatic interaction and hydrogen bonding likely contributes to the change of enthalpy that favors Z-conformation in the methylated states.

Blueberry Extract Reduces Oxidative Stress in Patients with Metabolic Syndrome

Oxidative stress is a health condition that could potentially harm the patient, and it is defined as the lack of balance between the production of oxygen free radicals, which rises, and antioxidant defenses, which are in decrease. Metabolic syndrome implies a variety of risk factors that are based on abdominal obesity and insulin resistance. Moreover, the importance of metabolic syndrome is emphasized due to the fact that it presents an increase in oxidative stress, which is produced by the sum of alterations that each risk factor produces within the metabolic syndrome pathology. Reducing oxidative stress in these patients is currently one of the most interesting challenges of cardiovascular and metabolic therapeutics, because it is a molecular biology alteration that is not generally diagnosed and, therefore, not treated. The increasing incidence of overweight and obesity cause an increase in the incidence of metabolic syndrome, thus turning into a huge problem that keeps growing at alarming proportions. This syndrome＇s incidence oscillates between 20 and 40%, depending on the gender, age, ethnic group, and diagnostic criteria used for the definition of the disease. Prospective studies show that metabolic syndrome doubles the risk of cardiovascular disease and causes a five-fold increase in the risk of developing type 2 diabetes. With the aim of decreasing the oxidative stress caused by metabolic syndrome, we investigated the effect of antioxidant protection in DNA repair and cell membranes through the use of blueberry extract, which is the fruit with the highest antioxidant capacity, in patients with metabolic syndrome. Thirty patients were studied for a period of 6 months of intervention, and it could be demonstrated that they showed a highly significant decrease in the damage produced to the DNA, which was measured by the urinary excretion of 8-hydroxy-2＇-deoxyguanosine （8-OHdG） and the damage caused to the vascular endothelium and cell membranes, which was measured through the urinary excretion of F2-isoprostane. Based on our knowledge, this investigation is the first one to show that lyophilized blueberry extract （BlueKing~） as a dietary supplement, with meals, is an additional therapeutic tool of great value for the treatment of oxidative stress through DNA and cell membrane protection in patients with metabolic syndrome.

离体大鼠肠道菌对6种皂苷类成分代谢研究

为考察离体大鼠肠道菌群对6种皂苷类成分代谢转化的情况,将6种皂苷类成分单体(三七皂苷R_1,人参皂苷Rg_1,人参皂苷Rg_2,人参皂苷Re,人参皂苷Rd和人参皂苷Rb_1)分别与离体大鼠肠道菌群在厌氧条件下孵育8,24 h,通过乙腈沉淀蛋白和乙酸乙酯萃取处理后,采用LC-Q-TOF-MS/MS对代谢产物进行定性分析。通过比较空白对照样品和加药孵育样品的总离子流以及各个色谱峰的准分子离子和多级碎片离子,分析各皂苷类成分在大鼠粪便中可能的代谢产物。结果发现,大鼠肠道菌群对6种皂苷类成分具有显著的代谢转化作用,三七皂苷R_1主要被代谢为5个产物,代谢途径为三七皂苷R_1→人参皂苷Rg_1→人参皂苷Rh_1和人参皂苷F_1→原人参三醇→脱氢原人参三醇;人参皂苷Rg_1主要被代谢为4个产物,代谢途径为人参皂苷Rg_1→人参皂苷Rh_1和人参皂苷F_1→原人参三醇→脱氢原人参三醇;人参皂苷Rg_2主要被代谢为2个产物,代谢途径为人参皂苷Rg_2→原人参三醇→脱氢原人参三醇;人参皂苷Re主要被代谢为4个产物,代谢途径为人参皂苷Re→人参皂苷Rg_2→人参皂苷F_1→原人参三醇→脱氢原人参三醇;人参皂苷Rd主要被代谢为4个产物,代谢途径为人参皂苷Rd→人参皂苷Rg_3和人参皂苷F_2→人参皂苷Rh_2→原人参二醇;人参皂苷Rb_1主要被代谢为5个产物,分别为人参皂苷Rb_1→人参皂苷Rd→人参皂苷Rg_3和人参皂苷F_2→人参皂苷Rh_2→原人参二醇。综上所述,6种皂苷类单体均能够被离体大鼠肠道菌群代谢,主要的代谢途径为脱糖基和脱氢。

高效液相色谱-电喷雾质谱分析川续断皂苷Ⅵ在大鼠体内的代谢产物

目的:研究川续断皂苷Ⅵ在大鼠体内的代谢。方法:采用高效液相色谱-电喷雾质谱检测大鼠灌胃川续断皂苷Ⅵ后血浆、尿液、胆汁和粪便中的代谢产物。使用Agilent公司Zorbax Extend-C18色谱柱(150 mm×4.6 mm,5!m),以0.05%甲酸乙腈-0.05%甲酸水为流动相梯度洗脱(0～12 min,25%B→27%B;12～30 min,27%B→50%B;30～40 min,50%B→80%B;40～45 min,80%B→90%B;45～50 min,90%B),进行色谱分离,并与电喷雾质谱联用,毛细管电压为2.50 kV,锥孔电压为35 V,干燥气流速为320 L.h-1以及离子源温度为120℃,根据全扫描检测正、负离子模式下获得的准分子离子峰推测化合物的分子量信息及其可能的化学结构。结果:在大鼠血浆、尿液、粪便和胆汁中均检测到原型成分,粪便中检测到川续断皂苷Ⅵ脱去1分子(RM1,RM2)、2分子(RM3,RM4)及全部糖基(RM5)的5个代谢产物,血浆和胆汁中检测出脱去2分子葡萄糖残基的代谢产物(RM4),此外在胆汁中首次发现川续断皂苷Ⅵ的羟基化产物(RM6)。结论:川续断皂苷Ⅵ在大鼠体内的转化主要是在肠道的脱糖基代谢及在肝脏的羟基化反应。

三七蒸制前后主要活性成分体内药动学及其抗血小板聚集活性比较研究

目的研究不同蒸制时间(0、2、4、8 h)三七主要活性成分变化、体内药动学和抗血小板聚集活性之间的关系。方法采用高效液相色谱(HPLC-UV)法分析三七蒸制前后主要活性成分变化;大鼠ig 500 mg/kg不同蒸制时间的三七样品液后于不同时间点眼眶取血,超高效液相色谱-串联质谱(UPLC-MS/MS)法测定三七蒸制前后主要皂苷的血药浓度,运用DAS 3.2.6软件计算各成分的药动学参数;采用PAPE-I型血小板聚集/血凝测定仪检测抗血小板聚集活性。结果蒸制过程降低了生物活性物质三七皂苷R_1及人参皂苷Rg_1、Rd、Rb_1、Re的水平,并产生了新的成分人参皂苷Rg_2、Rg_3、Rh_1、F_2、Rk_3、Rh_4。人参皂苷Rb_1、Rd的脱糖基代谢产物人参皂苷Rg_3与原型皂苷人参皂苷Rb_1比较,t_(max)略有降低,表明脱糖基之后吸收速度加快。蒸三七比生三七有更明显的抗血小板聚集活性,且随着蒸制过程的延长,抗血小板聚集活性更强。结论三七蒸制后,皂苷类成分脱糖基后可能产生活性更强的成分,某些脱糖基代谢产物更易吸收入血,可能导致抗凝血活性增强。

A novel mis-sense mutation (G1381A) in the G6PD gene identified in a Chinese man

Objective To detect new mutations among 29 glucose 6 phosphate dehydrogenase (G6PD) deficient individuals from Yunnan province Methods The nitroblue tetrazolium (NBT) method was used to screen G6PD deficient individuals Mutation was identified by single strand conformation polymorphism (SSCP), amplification created restriction site (ACRS), amplification refractory mutation system (ARMS) and DNA sequencing Results Among 29 cases, 18 cases of G1388A, 1 case of C1004A, and 1 case of G1381A were identified Nine cases remained to be defined The G1381A mutation is a novel mis sense mutation, with a substitution of threonine for alanine (A461T) The resultant G6PD had reduced enzymatic activity In addition, G1381A caused a restriction site of Stu I to disappear, providing a rapid method for the detection of this mutation Conclusion A novel mis sense mutation G1381A was found This mutation results in a substitution of threonine for alanine, producing enzyme with reduced activity The loss of the Stu I restriction site offers a rapid method for the detection of this mutation