1.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.
2.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.
3.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.
4.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.
5.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.
6.A preliminary study on the toxin of earthworm and the methods to take it off
Zuxun ZHANG ; Liming HE ; Peilan YU ; Dianer YANG ; Hua LIU
Chinese Journal of Biochemical Pharmaceutics 2001;22(3):112-113
Purpose The source of earthworm toxin, its ingredients and the methods to take it off were studied.Methods The irritable secretor was dialyzed, and then examined by UV-Vis spectrum instrument.The proteins was analyzed with electrophoresis. The toxicity was tested in vivo.Results The proteins of irritable secretor 0.242mg/kg iv cause a mouse to death.Many bands were separated from the proteins with SDS-PAGE, among which two kinds were the principal. Even with different stimulus the secretor of earthworm was identical in ingredients.Conclusion The results demonstrated that the earthworm proteins of irritable secretion had toxicity and the toxin could be removed from earthworm by adding NaCl or 40V electricity.
7.Molecular cloning of human FL gene and its expression in E.coli
Huaiping ZHU ; Zimin SUN ; Jian WANG ; Haiming DAI ; Jingsheng WU
Chinese Journal of Biochemical Pharmaceutics 2001;22(3):109-111
Purpose The aim is to obtain the cDNA sequence of encoding extramembrane human FL gene with high level expression in E.coli. Methods The primers were designed based on the known FL cDNA sequence. The total RNA was isolated from fetal liver cells , and then RT-PCR was performed. The fragment was cloned into pUC-18T vector, and further sequenced by automatic sequence analyzer. The gene was inserted into GST fusion expression vector between BamH Ⅰ and EcoR Ⅰ sites. The recombinant plasmid was transformed into E.coli strain DH5 α and induced with 1mmol/L IPTG.Results The 546bp DNA fragment was amplified by RT-PCR method from fetal liver cells and its sequence was identical to the published sequence encoding human FL. The expressed fusion protein, with molecular weight of about 22kD, was about 10% of the total bacteria protein by SDS-PAGE and densitometry analysis.Conclusion cDNA was cloned successfully. This study provided a basis for the further fundamental research and clinical application of FL.
8.Research of the activity and the inhibitors of cGMP specific PDE5
Shunzu HAO ; Yisen HUANG ; Juntian ZHANG
Chinese Journal of Biochemical Pharmaceutics 2001;22(3):144-146
Purpose The aim is to study the activity and the sildenafil selective inhibition of PDE5. Methods The PDE isoenzymes were purified from bovine penis corpus cavernosum tissue by FPLC system. PDE activity was assayed by using 3 H-cGMP as substrate, the PDE isoenzymes hydrolyzed it to 3 H-GMP, and 3 H-GMP was further hydrolyzed to 3 H-guanosine by 5′-nuclease of snake venom. Add scintillation cocktail to observe the PDE isoenzymes activity. The selective inhibitor sildenafil of different concentrations were used to observe the inhibition of PDE5. Data replotted according to procedure of Dixon plots.Results Three PDE isoenzyme peaks were purificated from bovine corpus cavernosum. The PDE of the third peak had the strongest activity of cGMP hydrolyzation which could be inhibited by sildenafil apparently.Conclusion Since PDE5 was mainly found in corpus cavernosum tissue of mammalian, and sildenafil was a selective inhibitor of PDE5. It was suggested that the third peak was PDE5. The result was in agreement with the article reported.
9.Synthesis of silver carboxymethyl chitosan and its experimental study on its bacteriostasis
Xuejun ZHAN ; Yuanzhen XIONG ; Zhe LIU ; Daze XIE
Chinese Journal of Biochemical Pharmaceutics 2001;22(3):142-144
Purpose The aim is to develop a synthesis method of the silver carboxymethyl chitosan and to study its bacteriostasis to Staphlococcus aureus(S.aureus),Pseudomonas aeruginusa(P.aeruginusa),Escherichia Coli(E.coli),Klebsiella pneumoniae(K.pneumoniae) and Proteus vulgaris(P.vulgaris).Methods Chitosan was modified by way of chemistry.The structure analysis of its derivate was analysed by infra-red absorption spectroscopy,using methods of dilution and concavering to study the bacteriostasis to some ordinary bacteria which cause infection in burn.Results The infra-red spectragram of the derivate showed the chitosan had been modified by chloroacetic acid.When the concentrationl of silver carboxymethyl chitosan was 1.028 mg/ml and the concentration of the five bacteria were 104CFU/ml,the rate of bacteriostasis was 88%、80.2%、75.3% to S.aureus、P.aeruginusa and E.coli respectively.The MIC of silver carboxymethyl chitosan was similar to that of AgNO3 when they act on S.aureus and P.aeruginusa,but the former was lower than later when acting on E.coli.Conclusion Silver carboxymethyl chitosan could inhibit some bacteria which caused infection in burn.It was a novel pharmaceutical in preventing and curing burn infection.
10.Microwave assisted extraction of superoxide dismutase from pig blood
Zhilong XIU ; Wei WANG ; Meiqing LU
Chinese Journal of Biochemical Pharmaceutics 2001;22(3):139-141
Purpose The aim is to establish a simple method to extract superoxide dismutase(SOD) from pig blood red cells.Methods A combined method-microwave heating and adding Cu2+ into the suspension-was used to extract SOD from the red cells. Results The undesired proteins were denatured by microwave heating and SOD was partly purificated in comparison with hemolysis. The processing time of the former was much shorter than the latter. Conclusion A novel cell disruption microwave heating, was a rapid and effective technique for the primary extraction of SOD.