3.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
4.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.
5.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.
6.Comparison of Three Methods for Culturing Rheumatoid Arthritis Synovial Fibroblasts
Sha MA ; Jun LIN ; Liang YU ; Song JIN ; Qin LI ; Hong ZHANG ; Hongtao FAN
Journal of Kunming Medical University 2016;37(6):52-56
Objective To compare the three kinds of methods for in vitro primary culturing of rheumatoid arthritis synovial fibroblast-like cells (RASFs), in order to get fast and effective culture methods. Methods Synovial tissue from RA synovial arthroscopic resection were treated by collagenase digestion method, modified tissue culture method, double enzyme digestion method respectively. By using an inverted phase contrast microscope, cell morphology and growth characteristics were observed and identified with vimentin staining. Trypan blue was used to count the number of living cells after culturing for 14d. Results The three primary methods could successfully isolate and culture RASFs, and RASFs met the morphological characteristics of vimentin-positive cells>95%, namely, the proportion of RASFs cell confluence was 70% after 16-20days by the collagenase digestion method,whose cell confluence proportion reached 95%after 4 weeks;and the cell confluence proportion was above 70%after 10-14days by modified tissue culture method,and the cell confluence proportion reached 85%after 4 weeks by the double enzyme digestion method. The comparison of the viable cells number cultured same number of synovial tissue by the three methods show the viable cells number cultured by the modified tissue culture method were (1.60±0.08) ×106, those by the collagenase digestion method were (1.41±0.08) ×106, those by the double enzyme digestion method were (1.19 ±0.05) ×106, which were with significant difference among them (P<0.05) .The comparison of incubation time of RASFs primary cells showed it took (267.50±16.58) mins by the collagenase digestion method, (183.75 ±11.08) mins by the double enzyme digestion method, and 149.10 ±13.71mins by the modified tissue culture method, with significant differences (P<0.05) .Conclusion Modified tissue culture for RASFs is an efficient and fast culture method, the number and purity of RASFs can meet the requirements for biology experiments.
7.siRNA inhibition of survivin gene expression in rheumatoid arthritis synovial fibroblast proliferation and apoptosis
Sha MA ; Jun LIN ; Song JIN ; Qin LI ; Hong ZHANG ; Liang YU
Chinese Journal of Biochemical Pharmaceutics 2015;(12):17-20
Objective To study the targeting survivin small interfering RNA ( siRNA ) to inhibit proliferation and apoptosis survivin gene expression in rheumatoid arthritis synovial fibroblasts ( RASFs) .Methods RA patients were isolated and cultured in vitro synovial fibroblasts ( RASFs) , designed and synthesized siRNA targeting survivin and negative control, by liposome transfection RASFs cell; real-time quantitative polymerase chain reaction (PCR) and Western blot RASFs detect mRNA expression and protein levels of survivin.Tetrazolium blue (MTT) assay of cell proliferation;TUNEL assay apoptosis.Results The experimental group compared with the negative control siRNA group and control group, 48h after transfection of synovial fibroblasts survivin mRNA and protein expression levels were significantly decreased ( P<0.05 ) .The experimental group compared with the negative control siRNA group and control group, synovial fibroblast proliferation after transfection significantly decreased ( P<0.05 ) . After the experimental group transfected 24h, 48h, 72h growth inhibition rates were (11.5 ±2.6)%, (26.2 ±3.4)%, (47.6 ±4.1)%, at 72 hours after transfection most significant.The rate of apoptosis in experimental group (23.87 ±1.6)%, significantly higher than the negative control group (9.72 ± 1.15)% and the control group (8.70 ±1.09)% (all P<0.05).Conclusion siRNA targeting survivin expression levels through reducing survivin, inhibit synovial fibroblast proliferation and promotes apoptosis.
8.The application of MR imaging in differentiating benign and malignant lymph node and assessing therapeutic effect
Caixian HAO ; Jun LIU ; Jin ZHONG ; Liang XU ; Xiuling ZHANG ; Yulin ZHANG ; Changqing ZONG ; Hui QIAO
Journal of Practical Radiology 2014;(10):1706-1709
Objective To approach the application value of diffusion weighted imaging with background suppression (DWIBS) combined with routine MR sequence in differentiating benign and malignant lymph nodes,and assessing therapeutic effect of lympho-genic tumors.Methods 48 patients with cervical lymph node enlargement who were confirmed by pathology and follow up under-went DWIBS and routine MRI examination,malignant lymph nodes were 83,benign lymph nodes were 79.16 patients with malig-nant lymph nodes were rechecked after radiotherapy and chemotherapy,Apparent diffusion coefficient (ADC)values of the solid part for lymph nodes were compared.Results More lymph nodes can be detected in DWIBS than conventional sequence.ADC values of the solid part for malignant lymph node(0.898±0.111)×10-3 mm2/s were lower than that of benign lymph node(1.043±0.106)× 10-3 mm2/s,there was significant difference between them (P<0.05).ADC values of all malignant lymph nodes after treatment (1.205±0.121)×10-3 mm2/s were significantly higher than that of pretherapy (0.883±0.090)×10-3 mm2/s (P<0.05).Conclu-sion DWIBS could more sensitively detect lymph node than conventional MR sequence.ADC value could provide some reference values for differentiating benign and malignant lymph nodes and assessing therapeutic effect.
9.Effects of agmatine on excessive inflammatory reaction and proliferation of splenic cells in mice with trauma
Zheng LIU ; Fengyan HOU ; He JIN ; Ya XIAO ; Xia FAN ; Xue YANG ; Jun YAN ; Huaping LIANG
Chinese Critical Care Medicine 2015;(2):110-114
ObjectiveTo observe protective effects of agmatine (AGM) on inflammatory response and spleen immune function in mice with trauma.Methods Forty-eight adult male C57BL/6 mice were randomly divided into three groups (n= 16 each), including control group, model group (bilateral femoral fracture and removal of 35% of the total blood volume), and AGM group (trauma/hemorrhage & AGM 200 mg/kg). Eight mice in each group were sacrificed at 3 hours and 24 hours, respectively, after modeling, and blood samples and tissue homogenate of spleen and liver were collected. The contents of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-1β) in serum and liver tissue were determined with enzyme linked immunosorbent assay (ELISA). Serum aspartate transaminase (AST), alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer. Spleen proliferation response stimulated with concanavalin A (ConA) was evaluated with methyl thiazolyl tetrazolium colourimetry (MTT).γ-interferon (IFN-γ) and IL-2 releases were determined with ELISA.Results Compared with control group, 3 hours after trauma/hemorrhage, the levels of serum TNF-α, IL-6, and IL-1β in model group were significantly elevated [TNF-α (ng/L): 145.38±31.50 vs. 23.06±11.14, IL-6 (ng/L): 496.94±50.76 vs. 47.13±17.47, IL-1β (ng/L): 321.31±43.02 vs. 29.25±16.24,allP< 0.01]. It was found that AGM treatment could alleviate the increase in serum pro-inflammatory mediators induced by trauma/hemorrhage, such as TNF-α (ng/L:111.56±25.47 vs. 145.38±31.50), IL-6 (ng/L: 412.56±44.33 vs. 496.94±50.76), IL-1β (ng/L: 273.38±45.25 vs. 321.31±43.02,P< 0.05 orP< 0.01). Twenty-four hours after trauma/hemorrhage, serum pro-inflammatory mediators were recovered to the levels in control group. There was no significant difference in TNF-α and IL-6 levels at 3 hours after trauma/hemorrhage among groups. Compared with control group, the expressions of liver TNF-α and IL-6 in model group were increased at 24 hours following trauma [TNF-α (ng/mg): 32.93±4.90 vs. 26.58±2.33, IL-6 (ng/mg): 11.20±1.66 vs. 8.38±0.89,bothP< 0.01]. However, AGM inhibited the level of TNF-α (ng/mg:28.92±3.16 vs. 32.93±4.90) and IL-6 (ng/mg: 9.03±1.28 vs. 11.20±1.66) in the liver as induced by trauma/hemorrhage (P< 0.05 andP< 0.01). At 24 hours after modeling, model group and AGM group had distinctly higher serum AST, ALT, LDH levels than those of control group [AST (U/L): 405.9±31.2, 245.7±22.1 vs. 128.2±15.9; ALT (U/L): 92.1±6.3, 51.6±5.0 vs. 30.1±3.2; LDH (U/L): 606.7±36.3, 478.7±25.3 vs. 384.0±16.6, allP< 0.01]. Nevertheless,the increase in serum AST, ALT and LDH was alleviated in AGM group (allP< 0.01). Meantime, trauma/hemorrhage produced a noticeable depression of proliferation of splenic cells and IFN-γ and IL-2 release stimulated with ConA compared with control group [proliferation rate: (40.97±4.13)% vs. (89.99±7.76)%, IFN-γ(ng/L): 91.6±12.3 vs. 353.2±21.5,IL-2 (ng/L): 53.4±6.4 vs. 91.0±12.2,allP< 0.01]. In contrast, AGM notably restored the capacity of proliferation response of splenic cells [proliferation rate: (74.86±5.75)% vs. (40.97±4.13)%, P< 0.01],enhanced the release of IFN-γ and IL-2 stimulated with ConA [IFN-γ (ng/L): 327.8±23.6 vs. 91.6±12.3, IL-2 (ng/L): 74.8±10.4 vs. 53.4±6.4, bothP< 0.01].Conclusion AGM can dramatically alleviate spleen immunosuppression, excessive inflammation and organ damage induced by trauma/hemorrhage.
10.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.