1.Research progress on plant resources distribution of vitexin and its pharmacological effects.
Cheng-bo GU ; Man CAI ; Xiao-han YUAN ; Yuan-gang ZU
China Journal of Chinese Materia Medica 2015;40(3):382-389
Vitexin, a naturally occurring flavone glycoside in plants, has many pharmacological effects, which is widely distributed in nature. This paper reviewed the research progress of the distribution of vitexin in the plant resources and its pharmacological effects, and summarized its application prospects, aiming to provide a useful reference for the development of vitexin-enriched plant resources.
Animals
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Antineoplastic Agents
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pharmacology
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Antioxidants
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pharmacology
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Apigenin
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pharmacology
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Humans
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Hypoglycemic Agents
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pharmacology
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Myocardial Infarction
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drug therapy
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Plant Dispersal
2.Expression and Identification Truncated Glycoprotein G of Bovine Respiratory Syncytial Virus in Escherichia coli
Jun-Ke FENG ; Fei XUE ; Jiao LI ; Li-Chuang ZU ; Yuan-Mao ZHU ; Xian-Gang REN ;
China Biotechnology 2006;0(12):-
Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.
3. Optimization of processing technology and study on antioxidant activity in vivo of superfine powder of Granati Pericarpium
Chinese Traditional and Herbal Drugs 2015;46(10):1454-1459
Objective: Processing technology and antioxidant activity in vivo of pomegranate superfine powder were investigated. Methods: Processing technology of superfine powder of Granati Pericarpium was optimized using single-factor tests and response surface methodology (RSM). The antioxidant effect in vivo of superfine powder of Granati Pericarpium with different particle, vitamin E, and normal saline were studied. Results: The optimally grinding conditions for Granati Pericarpium were grinding time of 25 min, grinding temperature of -17℃, and input quantity of 198 g. Under the optimally grinding conditions, the minimum particle size of superfine powder of Granati Pericarpium was 7.68 μm which was close to the predicted value of 7.96 μm. Therefore, the established regression model has good prediction capability. Experimental results show that the superfine powder and vitamin E groups compared with the coarse powder and blank groups could significantly improve the activity of SOD, CAT, and GSH-Px, and reduce the content of MDA in serum of mice. Conclusion: The ability of superfine powder group on the protection of membrane lipid peroxidation and scavenge free radical is quite superior. It indicates that the active ingredients in the superfine powder of Granati Pericarpium could be dissolved better and more quickly in solvent than those in coarse powder. Therefore, the superfine powder of Granati Pericarpium has the better anti-oxidant effect in vivo.
4.Study of curative effect of combined therapy in the elderly patients with oral squamous-cell carcinoma
Mao-Chang YE ; Lai-Ping WANG ; Rong-Xin LI ; Zu-Wu ZHU ; Mei-Mei CHEN ; Ran-Gang KANG ; Yuan ZHOU
Chinese Journal of Geriatrics 2001;0(03):-
0.05).There was significant difference in the 3-year survival rate between A and C group. Conclusions The 3-year survival rate was dramatically increased with combined therapy mainly by cisplatin, the dose of 60~80mg is tolerant for the elderly aged above seventy years, and perioperation complications can be cured.
5.Relationships between thiopurine methyltransferase gene polymorphisms and its enzymatic activity.
Xiao-Li MA ; Min-Yuan WU ; Ya-Mei HU ; Ping ZU ; Zhi-Gang LI
Chinese Journal of Oncology 2006;28(6):456-459
OBJECTIVETo investigate the relationship between the thiopurine methytransferase (TPMT) gene polymorphisms and its enzymatic activity, and to clarify the significance of TPMT activity and gene polymorphisms on individualized therapy with thiopurines.
METHODSThe TPMT activity and gene polymorphisms were determined in an unrelated population of 250 Chinese healthy blood donors, 100 cords blood and 280 patients with acute leukemia. The TPMT genotyping assay was based on polymerase chain reaction (PCR), restriction digestion of PCR products, denaturing high-performance liquid chromatography (DHPLC) and SNaPshot sequencing and direct DNA sequencing in the TPMT exon 5 (G238C), TPMT exon7 (G460A) and TPMTexon10 (A719G). Erythrocyte TPMT activity was measured by high-performance liquid chromatography (HPLC).
RESULTSThe frequency of TPMT polymorphism in 250 Chinese healthy blood donors, 100 cords blood and 280 patients with acute leukemia was low (3.5%), and all the varied alleles were TPMT* 3C (exon 10A719G). All of them were TPMT* 1/TPMT* 3C heterozygote. The TPMT activity was between 6 and 12 U. The activity in 95.1% was more than 12 U (13 - 32 U), while the activity in others (4.9%) was 6 - 12 U. TPMT activity and genotype were concordant. Of 630 subjects evaluated, TPMT activity of heterozygous individuals in Chinese healthy blood donors, cords blood and acute leukemia patients were 9.1 U, 9.3 U and 9.07 U, respectively, significantly lower than that in general population (17.6 U, 17.67 U and 18.6 U, respectively). In the samples analyzed, ten subjects with heterozygous phenotypes (6/15 acute leukemia children and 4/16 healthy blood donors and cords blood) did not have TPMT* 2, TPMT* 3A or TPMT* 3C. Therefore, other factors may affect on TPMT activity.
CONCLUSIONTPMT gene polymorphisms and its activity were concordant. The heterozygotes had low TPMT activity. Therefore, detection of TPMT genotype and its activity is useful. These findings hold a promise of improving the safety and efficacy of thiopurines therapy.
Acute Disease ; Child ; Chromatography, High Pressure Liquid ; Erythrocytes ; enzymology ; Exons ; Female ; Fetal Blood ; enzymology ; Genotype ; Humans ; Leukemia ; blood ; enzymology ; genetics ; Male ; Methyltransferases ; blood ; genetics ; Polymorphism, Single Nucleotide
6.Exploration of methodology for assay of single nucleotide polymorphism in thiopurine methyltransferase gene.
Xiao-Li MA ; Ping ZU ; Ya-Mei HU ; Min-Yuan WU ; Zhi-Gang LI ; Ding-Fang BU
Journal of Experimental Hematology 2003;11(5):458-463
The aim of the present study was to gain an insight into the thiopurine methytransferase (TPMT) genotyping assay, which was based on polymerase chain reaction (PCR), allele-specific PCR, restriction digestion of PCR products, denaturing high-performance liquid chromatography (DHPLC) and SNaPshot sequencing and in combination with direct DNA sequencing. Among the f our methods to test TPMT genetic SNPs based on PCR, allele specific PCR was not able to differentiate wild type from varied type. BsiYI, MwoI and AccI to digest PCR products were used so that SNP in TPMT exon 5, 7 and 10 tested. It showed that there were no differences between the results of digestion of PCR products and those of DNA sequence analysis. Therefore, this method was reliable. But some other methods were still needed to look for a compensation, because no restriction map changing resulted from the 2 SNPs in TPMT promotor was found. As to the results of DHPLC, those for the screening of TPMT exon-5 and -10 for SNPs were the same as restriction analysis of PCR products and direct DNA sequencing. But the variation of the heterozygotes in exon-7 was high, which was different from the results of direct DNA sequencing. After changing the Tm of DNA step by step, It was found that all the samples showed single peak when the temperature was 54 degrees C. But this result was unbelievable because a heterozygote in exon 7 as positive control could not be found. Therefore, it was necessary to test the sensitivity and accuracy of DHPLC, though DHPLC could be used as an effective method of SNPs screening. The results of the SNaPshot sequencing were also same as those of restriction analysis of PCR products and direct DNA sequencing. And the results showed that the bases of TPMT promoter -91 and -168 were G, instead of A and T. The results of the four methods to detect TPMT genetic SNPs based on PCR showed that SNPs analysis technique should be a combination of the techniques above-mentioned. One technique alone could not satisfy the need in clinics and research. The compensation of each other was very important.
Chromatography, High Pressure Liquid
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Exons
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Humans
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Methyltransferases
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genetics
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Polymerase Chain Reaction
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Polymorphism, Single Nucleotide
7.Isolation and purification of 10-hydroxycamptothecin and vincoside-lactam from Camptotheca acuminata seed by polyamide.
Wei-Guo SHI ; Yuan-Gang ZU ; Lei YANG ; Chun-Jian ZHAO ; Jia-Hui LI
China Journal of Chinese Materia Medica 2008;33(21):2486-2489
To investigate the technological parameters of the isolation and purification of 10-hydroxycamptothecin and vincoside-lactam from Camptotheca acuminata seed by polyamide. The static arid dynamic adsorption characteristics of 10-hydroxycamptothecin and vincoside-lactam on polyamide were studied, and the contents were determined by HPLC. The optimum parameters for adsorption were as follows: the contents of 10-hydroxycamptothecin and vincoside-lactam in the extracts were 0.189 g x L(-1) and 0.334 g x L(-1), respectively, pH 6, flow rate was 1.0 mL x min(-1), processing volume was 3 BV; for desorption: ethanol-water (60:40), flow rate was 1.0 mL x min(-1), 5 BV as an eluent. After treated with polyamide, the contents of 10-hydroxycamptothecin and vincoside-lactam were 17.52% and 32.87%, respectively, the recovery yields were 66.05% and 75.86%, respectively. Results showed that polyamide revealed a good ability to separate 10-hydroxycamptothecin and vincoside-lactam. Therefore, we concluded that results in this study may provide scientific references for the large-scale production of 10-hydroxycamptothecin and vincoside-lactam extracted from C. acuminata seed.
Camptotheca
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chemistry
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Camptothecin
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analogs & derivatives
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chemistry
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isolation & purification
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Hydrogen-Ion Concentration
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Nylons
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chemistry
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Seeds
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chemistry
8.Synthesis and antitumor activities of 10-hydroxy camptothecin derivatives.
Qing-Yong LI ; Yuan-Gang ZU ; Ying ZHANG ; Yu-Jie FU ; Qun LIU
Acta Pharmaceutica Sinica 2004;39(7):521-524
AIMTo find new anticancer drug based on the structure of 10-hydroxy camptothecin.
METHODSSeven camptothecin derivatives (3 -9) were synthesized and the antitumor activities of these derivatives were evaluated.
RESULTSStructures of seven new compounds were determined by 1HNMR, IR, MS. Seven compounds showed inhibitory effects on Hela, BEL-7402, 7901 cell lines in vitro. Especially, compound 4 showed high bioactivities to all of the tumor cells in vitro, its anticancer activity against human cervical carcinoma Hela was much higher than that of 10-hydroxy camptothecin.
CONCLUSIONSome compounds are worth further studying.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Camptothecin ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Humans ; Molecular Structure
9.Preparation, activity and targeting ability evaluation in vitro on folate mediated epigallocatechin-3-gallate albumin nanoparticles.
Yuan-gang ZU ; Shuai YUAN ; Xiu-hua ZHAO ; Yu ZHANG ; Xiao-nan ZHANG ; Ru JIANG
Acta Pharmaceutica Sinica 2009;44(5):525-531
To study the preparation, activity and targeting ability evaluation in vitro on epigallocatechin-3-gallate (EGCG) bovine serum albumin nanoparticles targeting to PC-3 cells, the folate mediated EGCG bovine serum albumin nanoparticles (FA-EGCG-BSANP) were prepared by desolvation process. The morphology and particle size of the nanoparticles were determined by atomic force microscope (AFM). HPLC was used to analyse the entrapment efficiency and drug loading rate of EGCG The amount of folate conjugation on the BSANP was determined by quantitative ultraviolet (UV) spectrophotometer analysis. The targeting ability to PC-3 was observed using laser scanning confocal microscope (LSCM) and fluorophotometer microscope. And the activity of FA-EGCG-BSANP was mensurated by MTT method. The morphology and particle size distribution of FA-EGCG-BSANP were uniform and even with the mean particle size of 200 nm. The entrapment efficiency and loading rate of EGCG were (81.5 +/- 1.8) % and (29.3 +/- 0.6) %, respectively, and the amount of folate conjugation was 18.363 microg x mg(-1) BSA. The FA-EGCG-BSANP uptakes by cultured PC-3 cells were 23.65 times the amount of EGCG-BSANP in a concentration dependant manner. The lethality of PC-3 cells treated with FA-EGCG-BSA was 82.8%, while those treated with EGCG and EGCG-BSANP were 58.6% and 55.1%, respectively. And lethality of PC-3 cells was positively correlated with the nanoparticles uptake amount. FA-EGCG-BSANP can significantly promote EGCG to PC-3 cells sites and improve their efficacy, which is considered to an experimental foundation for further research on its activity, targeting ability and metabolism in vivo.
Antineoplastic Agents, Phytogenic
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administration & dosage
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pharmacokinetics
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pharmacology
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Catechin
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administration & dosage
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analogs & derivatives
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pharmacokinetics
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pharmacology
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Cell Death
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drug effects
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Cell Line, Tumor
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Drug Carriers
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Drug Compounding
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Drug Delivery Systems
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methods
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Folic Acid
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administration & dosage
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chemistry
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pharmacokinetics
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Humans
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Male
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Nanoparticles
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Particle Size
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Prostatic Neoplasms
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metabolism
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pathology
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Serum Albumin, Bovine
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chemistry
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pharmacokinetics
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pharmacology
10.One-step multiplex RT-PCR for identifying common fusion transcripts in childhood acute lymphoblastic leukemia.
Xiao-Wen CHEN ; Fei-Qiu WEN ; Rong-Yu LV ; Min ZHANG ; Ying ZU ; Hui-Rong MAI ; Ying WANG ; Xiu-Li YUAN ; Chang-Gang LI ; Dong-Li MA
Chinese Journal of Contemporary Pediatrics 2015;17(4):332-336
OBJECTIVETo evaluate the efficiency of one-step multiplex RT-PCR for identifying four common fusion transcripts (TEL/AML1, E2A/PBX1, MLL/AF4 and BCR/ABL) in children with acute lymphoblastic leukemia (ALL).
METHODSTotal RNA was extracted from bone marrow samples of 76 children who were newly diagnosed with ALL between January 2003 and December 2010. These RNAs were analyzed for TEL/AML1, E2A/PBX1, MLL/AF4 and BCR/ABL by one-step multiplex RT-PCR or common nested-multiplex PCR. The PCR products were confirmed by DNA sequencing.
RESULTSTEL/AML1 was found in 12 cases (the length of products was 298 bp in 9 cases and 259 bp in 3 cases), E2A/PBX1 was found in 3 cases (the length of products was 373 bp), BCR/ABL was found in 1 case (the length of products was 2 124 bp), and MLL/AF4 was found in 7 cases (the length of products was 427 bp in 1 case and 673 bp in 6 cases) using one-step multiplex RT-PCR combined with DNA sequencing. The results were consistent with those using common nested-multiplex PCR.
CONCLUSIONSOne-step multiplex RT-PCR may be another alternative for detection of common fusion transcripts in children with ALL.
Child ; Child, Preschool ; Core Binding Factor Alpha 2 Subunit ; genetics ; Female ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Infant ; Male ; Multiplex Polymerase Chain Reaction ; methods ; Myeloid-Lymphoid Leukemia Protein ; genetics ; Oncogene Proteins, Fusion ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA