1.Determination of Secnidazole in Secnidazole Tablets by RP-HPLC
China Pharmacy 2001;0(10):-
OBJECTIVE:To determine the contents of secnidazole in secnidazol e tablets by RP-HPLC.METHODS:The chromatographic column was VP-ODS C 18 with acetonirile-0.05mol/L potassium dihydrogen phosphate-triethylamine(15∶85∶0.4)as the mobile phase,the detection wavelength was311nm and the flow rate was0.8ml/min.RESULTS:There was a good linear relationship when the sample size was within the range of0.166?g~0.832?g(r=0.9999),the average re?covery was99.84%(RSD=1.05%,n=15).CONCLUSION:The method is simple and accurate,sensible and reproducible,which can be used for the assaying of both the raw materials and preparations and the quality control of secnidazole tablets.
2. Preparation of polyelectrolyte layer-by-layer coated microgels for pulsed drug delivery
Academic Journal of Second Military Medical University 2011;32(11):1213-1217
Objective To Investigate the preparation method, the release profile and structure of the polyelectrolyte layer- by-layer coated chitosan-alginate microgels. Methods The cores of the microgels were prepared by a high voltage electrostatic system, and the semipermeable membrane outside the microgel was polyelectrolyte deposits on the core surface through electrostatic attraction. The influences of different ratios of materials on the expansion property and the in vitro cumulative release of the coated microgels were evaluated by a single factor experiment. Results The prepared polyelectrolyte-coated microgels were well-shaped, with a narrow range of diameter distribution. The lag time of in vitro release was 2. 67 h; the release was rapid after lagging, with the cumulative in vitro release being 72% within 3 h. Conclusion Polyelectrolyte layer-by- layer coated chitosan-alginate microgels can release payload in a pulsed fashion in vitro.
3. Pulsed release behavior of microgels coated layer-by-layer by polyelectrolyte in vivo and in vitro
Academic Journal of Second Military Medical University 2011;32(11):1218-1222
Objective To Investigate the in vitro and in vivo release of chitosan-alginate microgels coated layer-by-layer by polyelectrolyte self-assembly. Methods The cores of the microgels were made by gelatinization using electrostatic microencapsulated and coated by polyelectrolytes using electrostatic attraction. The effects of different layers and ratios of polymer on the in vitro lease of FITC-dextran were evaluated. Histrological examination was carried out to observe the in vivo release process by injecting the coated microgels into mice. Results The results showed that alginate and calcium chloride concentrations and polyelectrolyte layers markedly affected the lag time of pulsed release and the relasing speed after lagging. Conclusion The release of microgels coated layer-by-layer by polyllectrolyte can be controlled in vitro and can be observed in vivo; meanwhile, the microgels are safe and have good biocompatibility.
4. Three programmed systems for pulsed drug delivery
Academic Journal of Second Military Medical University 2010;31(7):782-786
Pulsed drug delivery (PDD), which can be released at well-defined time points as the therapy needs, can decrease the frequency and avoid taking drug at night, thus improving patient compliance. Here we introduce three kinds of programmed PDD systems independent of external chemical triggering; they are divided according to the triggering mechanisms, degradation-triggered PDD, osmotic pressure-triggered PDD, and both degradationi and osmotic pressure-triggered PDD. This paper reviews preparing technique, release mechanisms and influencing factors of the three PDD systems. The release profiles of pulsatile PDD can be regulated for different therapeutic needs, requiring no external triggers; especially that the PDD system triggered by both degradation and osmotic pressure has a bright future.
5.Construction and identification of the transgenic plant vector recombinant pBI-Eg95 plasmid of Echinococcus granulosus
Chinese Journal of Endemiology 2008;27(4):401-404
Objective To construct and identify the transgenic plant vector recombinant pBI-Eg95 plasmid of Echinococcus granulosus. Methods Total RNA was extracted from hydatid cyst protoscoleces of Echinococcus granulosus after sonication. A couple of specific primers were designed on the basis of known sequences of Eg95 gene. The desired gene was amplified by PCR technique from the cDNA, and then was cloned into the plant expression vector pBI121 to construct the recombinant pBI-Eg95 plasmid. The recombinant plasmid was electroporated into Agrobocterium tumefaciens (At) LBA4404 strain. The positive recombinant clones were confirmed by restriction endonuclease digestion and characterized by PCR. Results For RT-PCR, a specific band around 471 bp was amplified. The result of DNA sequencing of Eg95 showed the identity with the published sequence. The same band was obtained by restriction endonuclease digestion and PER from the plasmids of positive recombinant At(rAt). Conclusions The recombinant pBI-Eg95 plasmid was successfully constructed, and it provides the basis to further research of the transgenic plant vaccine of Echinococcus granulosus.
6. HPLC fingerprint of flavonoids in Dendrobium officinale from Zhejiang Province
Chinese Pharmaceutical Journal 2013;48(15):1261-1265
OBJECTIVE: To establish the HPLC fingerprint of flavonoids from Dendrobium officinale and screen important flavonoid components. METHODS: HPLC analysis was carried out on an XB C18 column(4.6 mm × 250 mm, 5 μm) with methanol and water containing 0.4% formic acid as mobile phase using gradient elution program. The detection wavelength was set at 335 nm. Total 11 batches of Dendrobium officinale and 15 batches of different species of Dendrobium were analyzed. The similarity evaluation system for chromatographic fingerprint of Traditional Chinese Medicine(2004AB) was applied to analyze the similarity. And principal component analysis (PCA) was used in data analysis. RESULTS: The HPLC fingerprint of flavonoids in Dendrobium officinale was established. Six peaks in this HPLC fingerprint were selected as common characteristic peaks, five of which were identified by HPLC-DAD-ESI2. Among the five peaks, four were flavone di-C-glycosides whose aglycone was apigenin, and the other one was naringenin. There were significant differences in the fingerprint chromatograms between Dendrobium officinale and different species of Dendrobium. PCA results indicated that apigenin-6,8-di-C-β-D-glucoside, schaftoside and naringenin were the most important components. CONCLUSION: The established method is highly characteristic and can provide reference for improving the quality control of Dendrobium officinale.
7.Integration and Expression of BGL1 Gene from Saccharomy-copsis fibuligera in Industrial Saccharomyces cerevisiae
Liang ZHANG ; Yan ZHOU ; Gui-Yang SHI ;
Microbiology 1992;0(03):-
The recombinant plasmid pPIC-gpd-bgl-hyg was constructed, which contained GPD2 promotor and terminator from industrial yeasts Saccharomyces cerevisiae, ?-glucosidase gene (BGL1) from Sac-charomycopsis fibuligera and hyg from hygromycin as the selected marker. With the yeast’s high efficiency of homologous integrated, the BGL1 gene was successfully integrated into industrial yeasts S. cerevisiae. The recombined yeast could grow on the cultures with the cellobiose as a sole carbon source, and the ?-glucosidase activity achieved 0.764 U/mL after 48 hours’ cultivation. In the experiments of VHG ethanol fermentation, the cellobiose concentration in broth of recombined yeast was 80% lower than that of indus-trial yeast.
8.Anti-infective immunity of recombinant Bifidobacteria bifidum-Eg95 vaccine of Echinococcus granulosus in mice
Chinese Journal of Endemiology 2012;31(6):608-612
Objective To investigate the protective immunity in mice immunized with recombinantBifidobacteria bifidum(Bb)-Eg95 vaccine of Echinococcus granulosus (Eg) and challenged with Eg protoscoleces.Methods Fifty-six female BALB/c mice 12-14 weeks old and weighed 20-25 g were vaccinated with the recombinant Bb-Eg95 vaccine subcutaneously,intramuscularly,intranasally and orally,respectively,with blank vector,Bb and medium of solution(MRS) as control,8 mice in each group.Mice were challenged with Eg protoscoleces on week 8 after immunization and killed on week 25 after infection.The weight of hydatid cyst was measured and the decreased larva rate was calculated.Sera were collected to determine the levels of IgE,IgG and its subclasses by enzyme linked immunosorbent assay(ELISA).Splenocytes were collected and cultivated to test the proliferation of splenocytes using methyltetrazolium (MTT) assay under EgAg and concanavalin A (ConA) stimulation.The results were compared with analysis of variance and the comparison between two groups was performed with LSD-t test.Results There was significant difference in the weight of hydatid cyst between groups (F =11.062,P < 0.05).Compared with MRS control group[(0.075 ± 0.019)g],the hydatid cyst weight decreased in subcutaneous group [(0.050 ± 0.013)g],intramuscular group[(0.050 ± 0.019)g],intranasal group[(0.028 ± 0.016)g] and oral group [(0.031 ± 0.018)g,all P < 0.01).Compared with subcutaneous and intramuscular groups,the hydatid cyst weight decreased in intranasal and oral groups(all P < 0.05).The decreased larva rate was inversely proportional to the weight of hydatid cyst.There was significant difference in the levels(obsorbancy,A) of IgG,IgG2a,IgG2b,IgG1,IgG3 and IgE between these groups(F =21.774,36.977,27.071,14.746,10.131,9.444,P < 0.05 or P < 0.01).Compared with MRS control group (0.015 ± 0.002,0.002 ± 0.001,0.003 ± 0.001),the levels of IgG,IgG2a and IgG2b increased in subcutaneous group(0.022 ± 0.004,0.007 ± 0.002,0.008 ± 0.002),intramuscular group (0.023 ± 0.003,0.008 ± 0.002,0.007 ± 0.002),intranasal group(0.032 ± 0.007,0.012 ± 0.002,0.013 ± 0.004)and oral group(0.028 ± 0.006,0.010 ± 0.003,0.010 ± 0.002,P < 0.05 or P < 0.01).Compared with subcutaneous and intramuscular groups,the levels of IgG,IgG2a and IgG2b increased in intranasal and oral groups(P < 0.05 or P < 0.01).Compared with MRS control group(0.009 ± 0.001,0.009 ± 0.002,0.009 ± 0.001),the levels of IgG1,IgG3 and IgE decreased in subcutaneous group(0.022 ± 0.004,0.007 ± 0.002,0.008 0.002),intramuscular group(0.004 ± 0.001,0.004 ± 0.001,0.004 ± 0.002),intranasal group(0.005 ± 0.002,0.005 ± 0.003,0.005 ± 0.002)and oral group(0.005 ± 0.001,0.004 ± 0.002,0.004 ± 0.003,all P < 0.01).There was significant difference in the proliferation of splenocytes in the supernatant of cultured splenocyte,of cultured splenocyte + EgAg and of cultured splenocyte + ConA(F =63.975,359.833,167.399,P < 0.01).There was significant difference in the proliferation of splenocytes inside groups(F =6741.955,4953.667,869.320,201.235,175.413,139.653,169.994,all P <0.01).Compared with the cultured splenocyte the proliferation of splenocytes increased in the cultured splenocyte +EgAg and splenocyte + ConA (all P < 0.01).Compared with the cultured splenocyte + EgAg,the proliferation of splenocytes increased in the cultured splenocyte + ConA(P < 0.01).Conclusion An effective and protective immunity is induced by the recombinant Bb-Eg95 vaccine of Eg in mice.
9.An association analysis of apolipoprotein E genotype and Alzheimer's disease in old population in Urumqi
Jiang HE ; Changwen ZHOU ; Junhao GUI
Journal of Clinical Neurology 1995;0(04):-
Objective To analyse the correlation of apolipoprotein E genotype and Alzheimer's disease(AD) in old population in Urumqi.Methods The polymerase chain reaction and restriction fragment length polymorphism(PCR-RFLP) techniques were used to detect the distribution of genotype, gene frequency of ApoE alleles in 60 cases with sporadic AD and 90 normals as control.Results The frequency of ApoE?2, ApoE?3, ApoE?4 was 6.67%, 75.83% and 17.50% respectively in AD group, while in control group, it was 13.33%, 79.45% and 7.22% respectively. The frequency of ApoE?4 allele in AD group was higher than that in control group( P
10.Research about NNMT gene in cancer
Weimin ZHOU ; Kuangbiao ZHONG ; Ming GUI
Journal of International Oncology 2013;40(10):732-735
Nicotinamide N-methyltransferase (NNMT) is a S-adenosyl-L-methionine (SAM) dependent cytoplasm enzyme,which plays a vital role in the biotransformation and detoxification of many drugs and xenobiotic compounds.Recent studies have revealed abnormal expression of NNMT in many tumors,which may contribute to tumorigenesis and tumor development and radiotherapy or chemotherapy resistance.