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Expression of Glu、mGluR 5 and EAAT 1 in bone tissues of ovariectomized osteoporotic rats and the effects of Total Flavonoids of Rhizoma Drynariae on it

Ji FANG ; Li YANG ; Jiazhen SHEN ; Heng LIANG ; Ronghua ZHANG

Chinese Journal of Biochemical Pharmaceutics.2014;(2):10-12,16.

Objective To observe the expression of Glu、mGluR 5 and EAAT 1 in bone tissues of ovariectomized osteoporotic rats and the effects of Total Flavonoids of Rhizoma Drynariae (TFRD) on it. Methods 45 SPF 3-month-old Sprague-Dawley (SD) female rats were randomly divided into sham operation (Sham, n=15) group and ovariectomized (OVX, n=30) group. The osteoporotic(OP) model was established by bilateral ovariectomy, 14 weeks later, we measured bone mineral density(BMD) by dual-energy X-ray and determined that OP model was successfully replicated, OVX group rats were then divided into OVX group (n=15) and OVX+TFRD group (n=15). The OVX+TFRD group was given TFRD for 12 weeks. Glutamate (Glu), metabotropic glutamate receptor 5 (mGluR 5), and Glutamate/Aspartate Transporter (GLAST/EAAT 1)’s expression of femur was examined in order to clarify the characteristics of bone glutamate signaling pathway and the effects of TFRD on it. Results Glu and ionotropic receptors mGluR 5 mainly distributed in bone marrow cells and osteoblasts closed to the bone marrow cavity walls. There were no significant differences in Glu expression among Sham group, OVX group and OVX+TFRD group. The mGluR 5 expression of OVX+TFRD group was significantly higher than that of Sham group and OVX group(P=0.009), while no significant difference was found between the latter two groups. In addition to large distribution in bone marrow cells, small amount of transporter EAAT 1 was noted to express in bone cells of the bone lacunae. There were no significant differences in EAAT 1 expression among the three groups. Conclusion In bone glutamate signaling pathway, this study demonstrated that TFRD could significantly improve the ionotropic receptor mGluR 5’s expression, but had no inlfuence for Glu and EAAT 1.

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Protective effects of resveratrol on acute lung injury induced by lipopolysaccharide in mices

Shujian XU ; Xingxing HUANG ; Ruifang ZHU ; Shuqin YU

Chinese Journal of Biochemical Pharmaceutics.2009;30(6):375-378,382.

Purpose To investigate the effect of resveratrol on acute lung injury induced by lipopolysaccharide (LPS) in mice.Methods Acute lung injury (ALI) was induced by LPS in mice. The changes of lung airway inspiratory resistance (Ri), expiratory resistance (Re), and dynamic lung compliance (Cdyn) were observed with pulmonary function test apparatus. Brochoalveolar lavage fluid (BALF) was collected, and the concentrations of interleukin 1β (IL-1β)、interleukin 6 (IL-6) and tumor necrosis factory α (TNF-α) were measured by ELISA. The ratio of wet to dry weight was calculated to assess lung edema. Pulmonary vascular permeability was examined with injection Evans blue to judge the destructive extent of alveolar epithelial cell and endothelial. Pathological section was made, and the histopathological change was observed with light microscope.Results Resveratrol can inhibit the elevation of Ri and Re, and the descent of Cdyn. Simultaneously, resveratrol reduced the concentration of IL-1β, IL-6 and TNF-α,as well as the wet to dry weight ratio and the pulmonary vascular permeability significantly. Furthermore, it also could attenuate the lung injury on histopathology.Conclusion The results show that pretreatment with resveratrol has a protective effect on ALI induced by LPS. The ultimate inhibiting and release of inflammatory factors were involved in the mechanism of the effects.

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The antiviral effect of interleukin 29 against hepatitis B virus in vitro

Yu CHAI ; Xin LUO ; Hailiang HUANG ; Daojun HU ; Zize TAO ; Shengquan ZHANG

Chinese Journal of Biochemical Pharmaceutics.2009;30(6):368-370,374.

Purpose To explore the antiviral effect of interleukin 29(IL-29) on hepatitis B virus in vitro.Methods To study the antiviral effect of IL-29 against hepatitis B virus by the amount of HBV mRNA detected .Through the quantity of mRNA translated from genes of MxA,2′,5′-OAS,PKR and RNase L as well as the signal pathway induced by IL-29,we used RT-PCR and Western blot to discuss the anti-hepatitis B virus mechanism which was stimulated by IL-29.Results The amount of HBV mRNA in HepG2.2.15 cells was reduced by stimulation of IL-29.The expression of MxA and 2′,5′-OAS was up-regulated,as well as P-ERK and P-AKT were activated by IL-29.Conclusion These findings showed that IL-29 had obvious antiviral activity towards HBV in HepG2.2.15 cells.

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Preparation and identification of anti-HIV-1 gp120 monoclonal antibody

Yongchen ZHANG ; Jian WEN ; Liang ZHANG ; Lei ZHAO

Chinese Journal of Biochemical Pharmaceutics.2010;31(1):26-28,31.

Purpose To Prepare anti- HIV-1 gp120 monoclonal antibodies and to identify the specificity of antibodies in order to provide technique for preparing HIV remedial antibodies. Methods The gene fragment of HIV-1 gp120 was connected to PEGX-4T-2 prokaryotic expressing vector. The vector was cut by enzyme. GST-HIV protein was expressed by E. coli XL1-blue. The BALB/c mice were immunized with purified GST-HIV antigen, and then the fusion of mice spleen cell and myeloma cell SP2/0 was executed as the routine cell-fusion technique. Positive cells were screened by Indirect ELISA. Immuno-blotting assay and Western blot identified the specificity of antibodies. Results External gene section from the recombinant plasmid by sequencing showed the same size of HIV-1 gp120 gene sequences. An external expressed protein band of 32 KD was obtained after purified protein SDS-PAGE electrophoresis. It indicated that six strains of hybridoma cells secreting special monoclone antibodies had been obtained. ELISA results showed that six strains monoclonal antibodies only reacted to HIV-1 gp120 antigen. Western blot results showed that a band with molecular weight 32 kDa was obtained, which could interact with HIV-1 gp120 monoclonal antibody. Conclusion Six strains of hybridoma cells secreting special monoclone antibodies had been obtained. The prepared monoclonal antibodies have established a basis for HIV remedial antibody.

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Study on preparation technique of enteric coating Potassium Sodium Dehydroandroan Drographolide Succinate pellets

Miao XU ; Yan LI ; Lei SHENG ; Fengyan WANG ; Yang SHI ; Guihua HUANG

Chinese Journal of Biochemical Pharmaceutics.2010;31(1):6-9,13.

Purpose To build up the Potassium Sodium Dehydroandroan Drographolide Succinate enteric coating sustained-release pellets delivery system by aqueous dispersion coating technique. Methods 1. Adopting MCC as vehicle, SiO_2 as antisticking agent, appropriate amount of 40% ethanol, preparing the core of pellets by extrude-spheronization method and the formulation and manufacturing process were optimized by orthogonal design. 2. Preparing the coating material with Eudragit L 30 D, which is used as pore-forming agent, EC as blocker and PEG6000 as plasticizer. The pellets were coated by fluid-bed coating method. Results 1. The optimal formulation and manufacturing process of pelltes' core were as follows: drug: MCC: SiO_2 = 7:7:5, extruding rate: 1 080 r/min, rounding rate: 960 r/min, rounding time: 5 min. 2. After the addition of EC: Eudragit L = 35:65, the plasticizer is 1.71 % and weight gain is 5% . The release in the gastric model fluid(pH 1.0) < 10% , and complete release ( > 80% ) in the enteric model fluid(pH 6.8) was in two hours. The release behavior accords with the regulations on the release rate of enteric preparation in ChP. Conclusion By adjusting the formulation and the parameters of the process of pellet and coating, we can make enteric coating Potassium Sodium Dehydroandroan Drographolide Succinate sustained-release pellets. All this accords with the regulation of pharmacopedia in vitro release.

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Study on the sterols of Hericium erinaceus extracts

Jieli LI ; Ling LU ; Yihua CANG ; Chuanchao DAI ; Jianyong QIU ; Kun CHEN

Chinese Journal of Biochemical Pharmaceutics.2001;22(3):124-126.

Purpose　The aim is to compare the sterols in Hericium crinaceus ethanol extract and water extract from solid fermented hyphae and to study the pharmaceutical chemical basis of the different medicinal effects. Methods　The nonsaponifiable lipids were isolated by saponification.The sterols were then detected by TLC and RP-HPLC.Results　The content of sterols in ethanol extract was found to be higher than that in water extract.And one type of sterols from Hericium erinaceus hyphae was identified as ergosterol.Conclusion　Due to ergosterol′s multifunction in biological activities,it may well be one of the active components of hyphae.And higher content of lipids, especially sterols may be one of the reasons for the better medicinal effect of ethanol extract than water extract.

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Laboratory study on reproducibility of no-albumin human plasma by rivanol precipitation Ⅰ.Purification or isolation of protein C and protein S

Yonghui TAO ; Jian JIN ; Bocheng WANG ; Manda ZHANG ; Changgeng RUAN

Chinese Journal of Biochemical Pharmaceutics.2001;22(3):121-124.

Purpose　The aim is to isolate and purify Protein C(PC) and Protein S(PS) from no-albumin human plasma by rivanol precipitation. Methods　The isolated and purified steps included adsorption onto and elution from barium, salting-out, ion-exchange chromatography, affinity chromatography,preparative isoelectric focusing and so on. Results　The molecular weights of the obtained PC and PS were (61±0.9)kD and (83±0.8)kD, respectively, the isoelectric point, 4.70±0.03 and 5.20±0.03,and the yield, 28.3% and 12.6%. The purified PC and PS were shown to be highly homogeneous by capillary zone-electrophoresis(CZE), and rich in Glu, Leu and Gly or Asp, Glu and Leu respectively. Conclusion　 The methods could be used for large-scale isolation and purification of PC and PS from no-albumin human plasma.

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Preparation and preliminary pharmacodynamic study of rectal suppository of low molecular weight heparin

Guangxi ZHAI ; Hengchang ZANG ; Yuanjun ZHONG

Chinese Journal of Biochemical Pharmaceutics.2001;22(3):119-120.

Purpose　The aim is to investigate the rectal suppository of low molecular weight heparin（LMWH）.Methods　The suppository was prepared by way of heating and fusion，the drug release property was studied with the release test in vitro and the absorption of LMWH into blood was tested by the changes of rabbits blood coagulation time. Results　The drug release property in vitro followed Fick′s diffusion equation and LMWH could enter the blood through rectal medication. Conclusion　The absorption from the rectum was another route of LMWH into the blood.

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The immunomodulatory effect of polysaccharide from brown seaweed

Linsheng ZHAN ; Xinsheng ZHANG ; Xiaohong WU ; Yingli WANG ; Zhixian WANG

Chinese Journal of Biochemical Pharmaceutics.2001;22(3):116-118.

Purpose　The aim is to investigate the effects of BSP on immunological function in normal and immunosuppressed mices.Methods　Thymus and spleen indexes, peripheral blood WBC number,the lymphocyte proliferation response, IL-2 production and serum and splenocyte hemolysin contents were measured after intraperitoneal injection of BSP in normal and immunosuppressed mices.Results　(1)BSP 100 mg/（kg*d）×10d significantly increased the thymus and spleen indexes and peripheral blood WBC number in immunosuppressed mice.The thymus and spleen indexes in normal mice was also increased. In addition,BSP markedly improved T,B lymphocyte proliferation responses and IL-2 production in normal and immunosupressed mices.(2) BSP improved the serum and splenocyte hemolysin contents in normal and immunosuppressed mice. Conclusion　It was suggested that BSP was a kind of immunomodulator, and could improve the immunological function of normal and immunosuppressed mices.

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Study on anti-virus effects of egg yolk immune solution of anti-influenza virus

Deyuan CHEN ; Gang XU ; Mingrong XIE ; Shuqi HUANG ; Ping ZHU

Chinese Journal of Biochemical Pharmaceutics.2001;22(3):114-116.

Purpose　The aim is to find out an effective preventing influenza immune preparation. Methods　Egg yolk immune solution of anti-influenza virus(FM1 stem) was prepared from egg of the hens that had been successfully immunized with influenza virus (FM1 stem). Its effects of anti-virus were observed through animal experiment.Results　When the mice of control group began to die the average daily drink quantity of the mice of normal saline control group, egg yolk immune solution control group, immediate preventive group was (3.06±0.86), (2.93±1.47) and (3.99±0.21)ml(P＜0.05)respectively. The average body weights of the mice of these three groups were (15.85±2.70),(14.58±1.92) and (18.27±1.71)g(P＜0.05)respectively. Their mortality was separately 53.84%,69.23% and 3.84%(P＜0.01). The antibody positive rate of survived mice′s serum (1∶10 diluted) was separately 100%, 100% and 0%(P＜0.01). Conclusion　The anti-virus effects of egg yolk immune solution of anti-influenza virus was powerful. The result of preventing mice′s influenza was remarkable.

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