1.Studies on Establishing and Optimizing Conditions of Concentration Virus in Water Body
Bao-Jin PAN ; Jun-Yi LIU ; Mei-Liang WEI ;
Microbiology 1992;0(06):-
In this experiment six methods,calcium chloride(CaCl_(2)) precipitation,polyethylene glycol(PEG,pH7.0) precipitation,polyethylene glycol(PEG,pH11.5) precipitation,aluminum chloride(AlCl_(3)) precipitation,Amicon Utcra centrifugal filter devices and cellulose nitrate membrane were used to concentrate the vaccine poliovirus type 1(PV_(1)) added to water body;experimental conditions for concentration were selected and optimized.The results showed that two methods,CaCl_(2)and PEG(pH 7.0) precipitation were suitable for concentrating virus in large volumes of water while amicon utcra centrifugal filter devices for small ones.The virus recovery of the three methods reached a 100% rate.
2.Beneficial effect of ecdysterone on rat myocardial infarction induced by coronary occlusion
Xu WU ; Jun JIN ; Ziwen LIANG ; Fusheng SHI ;
Chinese Traditional and Herbal Drugs 1994;0(08):-
Object To explore the beneficial effect of phytoecdysone (EDS) on myocardial infarction and its mechanism of action Methods Rat myocardial infarction model was prepared by ligating the left anterior descending coronary artery, and EDS was injected ip for seven consecutive days Serum creatine phosphokinase (CPK) glutamic oxalacetic transaminase (GOT), lactic dehydrogenase (LDH) activities, infarct size(IS), coronary blood flow, capillary vessel density and vascular endothelial growth factor (VEGF) expression were determined Results 0 5, 5, and 50 mg/kg of phytoecdysone were able to effect the activities of serum CPK, GOT, LDH in a dose depending manner with an optimal effect for improving cardiac zymogram at the dose of 5 mg/kg ip At this dosage EDS can markedly reduce IS, increase coronary blood flow, capillary vessel density and the expression of VEGF Conclusion ESD can alleviate myocardial infarction symptoms The mechanism of such beneficial effect may due to its ability to promote VEGF expression regeneration of capillary vessels and increase coronary blood flow
5.Effect of serum B lymphocyte stimulator on patients with systemic lupus erythematosus
Liang XU ; Zhi LI ; Jun ZHANG ; Jin-Ming LU ;
Chinese Journal of Rheumatology 2003;0(07):-
0.05).Conclusion High level of Blys could be observed in serum of patients with SLE and somewhat correlated with the lupus acitivity,although the level of Blys in serum can not reflect morbility and mortality of SLE patients.
6.Phase Transfer Bioconversion of Penicillin G into 6-APA by Immobilized Penicillin Acylase in Recycling Aqueous Two-phase Systems With Light-pH Sensitive Copolymers
Ke-Ming JIN ; Xue-Jun CAO ; Ying-Ping ZHUANG ; Ju CHU ; Si-Liang ZHANG ;
China Biotechnology 2006;0(10):-
Immobilized penicillin acylase was used for bioconversion of penicillin G into 6-APA in aqueous two-phase systems consisted of a light-sensitive polymer PNBC and a pH-sensitive polymer PADB.Partition coefficients of 6-APA was found to be:about 5.78,in the presence of 1% NaCl.Enzyme kinetic showed that reaction reached equilibrium at 7h or so.The 6-APA mole yields were 85.3%(pH 7.8,and 20 ℃) and this value was about 20%higher than control in reaction of single aqueous phase buffer.Partition coefficient of penicillin G(Na) washardly changeable,while partition coefficient of product,6-APA and phenylacetate acid was significantly changeable.Reason is due to Donnan effect of phase systems andhydrophobicity of products.The change of partition coefficients of products also affects bioconversion yield of products.In the aqueous two-phase systems,substrate,penicillin G,products 6-APA and phenylacetate acid are biased in top phase,while immobilized penicillin acylase is completely partitioned in bottom.Substrate,penicillin G enters into bottom phase,and it is catalyzed into 6-APA and phenylacetate acid,then the products enter into top phase.Finally,inhibition of substrate and products is removed to result in improvement of products yield.Moreover,immobilized enzymehashigher efficiency than immobilized cells and occupy smaller volume.Comparing with free enzyme,immobilized enzymehashigher stability,longer use life,completely partitioned in bottom phase and recycle.Bioconversion in two-phase systems using immobilized penicillin acylase showed outstanding advantage.The light-sensitive copolymer forming aqueous two-phase systems could be recovered by laser radiation at 488 nm or filtrated 450 nm light,while pH-sensitive polymer PADB could be recovered by isoelectric point(pH 4.1).The recovery of the two copolymers was 95%~99%.
7.PDT combined with intravitreal injection of ranibizumab treatment for age - related macular degeneration with CNV
Xue-Yan, YANG ; Jin-Xian, HE ; Xian-Jun, LIANG ; Sheng, ZENG
International Eye Science 2015;(4):659-661
AIM: To observe the curative effect of PDT combined with intravitreal injection of ranibizumab treatment for age-related macular degeneration with choroidal neovascularization ( CNV) .
METHODS:In accordance with the inclusion criteria, by indocyanine green choroidalangiography ( ICGA ) and optical coherence tomography ( OCT ) examination confirmed the diagnosis of macular CNV in 27 patients (27 eyes), treated with PDT 3 ~ 7d professional intravitreal injection of ranibizumab. At 1, 3, 6mo after treatment, the results of best corrected visual acuity (BCVA), FFA, ICGA, OCT examination and complications were observed.
RESULTS: The BCVA improved in 17 eyes ( 63%) , stable in 6 eyes ( 22%) , and decreased in 4 eyes ( 15%) . Before treatment, the average leakage area was 1 005. 69±105. 47μm, it were 875. 54 ± 103. 27μm, and 423. 37 ±79.68μm at 1 and 3mo after treatment, there were significant differences compared with before treatment ( P<0. 01). Average central macular thickness of retina before treatment was 485. 58±122. 59μm, and 398. 84±105. 32μm, 297. 74±89. 18μ m at 1 and 3mo after treatment, there were significant differences compared with before treatment( P<0. 01).
CONCLUSION: The method that PDT closed CNV combined with intravitreal injection of ranibizumab can effectively block angiogenesis recurrence, reduce the number of PDT treatment again and complications, improve the therapeutic effect.
8.Effects of hIFN-λ1 recombinant adenovirus on proliferation of gastric adenocarcinoma cell line and its mechanism
Jun LIU ; Yulan YAN ; Jin ZHANG ; Yang LIU ; Jie ZHANG ; Bing LIANG ; Xuefeng BU
Chinese Journal of Microbiology and Immunology 2013;(7):507-512
Objective To explore the effects of Ad-hIFN-λ1 recombinant adenovirus on prolifera-tion of gastric adenocarcinoma cell line SGC-7901 and its mechanism .Methods Ad-hIFN-λ1 recombinant adenovirus and empty plasmid Ad-LacZ were respeetively transfated to human gastric adenocarcinoma SGC-7901 cells.The proliferation of transfected cells was detected by MTT assay .IFN-λ1 gene expression at mR-NA and protein levels in the cells were measured by RT-PCR analysis and immunofluorescence assay , re-spectively .Tunel assay and flow cytometry were used to analyze the rate of cell apoptosis .Results The proliferation of gastric adenocarcinoma SGC-7901 cells were significantly inhibited with Ad-hIFN-λ1 inter-vention in accordance with highly expressed IFN-λ1 at mRNA and protein levels , respectively .The apoptosis rate of Ad-hIFN-λ1 transfected cells was markedly higher than that of the empty plasmid Ad-LacZ group and PBS blank control group .Conclusion The expression of hIFN-λ1 could be detected after transfection of Ad-hIFN-λ1 recombinant adenovirus into gastric adenocarcinoma SGC-7901 cells.Ad-hIFN-λ1 could signifi-cantly inhibit the proliferation of gastric adenocarcinoma SGC-7901 cells by promoting the apoptosis of cancer cells.
9.Diterpene ginkgolides meglumine injection inhibits oxidative stress induced by oxygen-glucose deprivation by activating Akt/Nrf2 pathway in SH-SY5Y cells
Zhiquan JIN ; Zhiliang XU ; Qiu LIU ; Jun ZHOU ; Liang CAO ; Gang DING ; Zhenzhong WANG ; Wei XIAO
Chinese Journal of Pharmacology and Toxicology 2017;31(1):65-72
OBJECTIVE To investigate the protective effects and mechanism of diterpene ginkgolides meglumine injection (DGMI) against oxidative stress induced by oxygen-glucose deprivation (OGD) in SH-SY5Y cells. METHODS SH-SY5Y cells were divided into five groups: normal control, model control (OGD group) and drug(25 mg · L- 1) administration groups including DGMI group, extract of ginkgo biloba leaves injection group (EGBLI) and lactones ginkgo biloba injection group (LGBI). The cells suffered from oxygen-glucose deprivation (OGD) for 4 h, followed by reoxygenation with drugs for 6 h. Then, cell viabilities were detect using CCK-8 assays, reactive oxygen species (ROS) levels using fluorescence probe DCFH-DA and superoxide dismutase (SOD) activities using WST-1 test. Western blotting was used to detected protein levels of hemeoxygenase-1(HO-1), NAD(P)H, quinone oxidore?ductase l (Nqo1), protein kinase B (Akt), phosphorylated Akt (p-Akt), nuclear factor-E2-related factor2 (Nrf2) and phosphorylated Nrf2 (p-Nrf2). The cells were induced by OGD for 4 h, followed by reoxygen?ation and DGMI for 1 h, combined with different concentrations of PI3K inhibitor (LY294002) (at the final concentration of 12.5, 25 and 50 μmol · L-1) before the protein levels of AKT, p-AKT, Nrf2 and p-Nrf2 were detected by Western blotting. RESULTS SH-SY5Y cells induced by OGD for 4 h resulted in an increase in ROS(P<0.01), but a decrease in cell viabilities(P<0.01), SOD activities(P<0.01), and antioxidant protein levels ( Akt, p-Akt, Nrf2, p-Nrf2, HO-1 and Nqo1) (P<0.01). Compared with OGD group, treatment with reoxygenation and drugs (DGMI,EGBLI and LGBI respectively) for 6 h resulted in a decrease in ROS (P<0.01), but an increase in cell viabilities, SOD activities and antioxidant protein levels of p-Nrf2, HO-1, Nqo1 and p-Akt(P<0.05,P<0.01). DGMI group showed the best efficiently. Moreover, after OGD for 4 h, compared with DGMI group, combining reoxygenation and DGMI with LY294002 for 1 h resulted in a concentration-dependent inhibition of the protein levels of p-AKT and p-Nrf2(P<0.01). CONCLUSION DGMI 25 mg · L-1 can inhibit oxidative stress in SH-SY5Y cells induced by OGD by increasing the activity and expression of Nrf2 through PI3K/Akt pathway, which may be one of the mechanisms by which DGMI protects neurons from stroke.
10.Anti-tumor effect and its mechanism of co-administration of fusion proteins hVEGF121/βhCG and mGM-CSF/βhCG
Liangliang JING ; Zitao MIAO ; Manman LI ; Jia YE ; Liang JIN ; Rongyue CAO ; Jun LONG
Journal of China Pharmaceutical University 2017;48(1):102-109
This study aimed at investigating the inhibitory effects and the anti-tumor mechanisms of co-adminis-tration of fusion proteins mGM-CSF/βhCG ( GC ) and hVEGF121/βhCG ( VC ) on RM-1 prostatic cancer and B16 F10 melanoma in the C57 BL/6 J mouse model. Two recombinant stains containing pET-28 a-mGM-CSF-X10-βhCGCTP37 and pET-28 a-VEGF-M2-X10-βhCG-CTP37 were induced by lactose to express fusion proteins. The fusion proteins were separated and purified to prepare the anti-tumor protein vaccines ( VC protein vaccine and GC protein vaccine) , which were then mixed to prepare a combined protein vaccine named VGC protein vac-cine. The prostatic cancer and melanoma tumor-bearing mice C57 BL/6 J were immunized with described vac-cines, then the growth of each tumor was measured;splenocyte proliferation of immunized mice was detected;and the cytotoxic effects of the vaccine on tumor cells were tested. After that, the in vivo concentrations of IFN-γ and anti-hVEGF antibodies were investigated by ELISA. The difference between each experimental group and normal saline group ( NS) was statistically significant in both tumor-bearing mouse models ( P <0. 05) respectively. Besides, VGC group exhibited significantly better anti-tumor effect compared with the GC and VC groups with the anti-tumor rate ( 41. 7 ± 0. 83)% and ( 46. 4 ± 1. 27)% for prostatic cancer and melanoma tumor, respectively. The co-administration of the two proteins, VC and GC, could inhibit the growth of RM-1 prostatic tumor and B16F10 melanoma effectively via anti-tumor immunity and anti-tumor angiogenesis.