肺部肿瘤组织与正常肺组织微量元素含量测定

体内微量元索的变化在癌变的发生。发展过程中的作用,已越来越引起人们的重视。Maqgalioth报道了部分正常组织与恶性组织的铜、锌水平,但有关肺癌组织及其正常(周边)组织的微量元素含量还未见报道。本文对六例肺癌患者肺部癌组织与其正常组织中的铜、锌、铁、锰、钙的水平进行了探讨。

Purification,Characterization and in vitro Anthelmintic Activity of a Neutral Metalloprotease from Laccocephalum mylittae

A neutral metalloprotease was purified from the cultured mycelia of Laccocephalum mylittae,an effective medicinal fungus widely used in anthelmintic therapy.The protease was purified to homogeneity with 31.85-fold purification and a final yield of 21.76%.The subunit molecular weight of the protease is about 40000 estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).The optimum reaction pH and temperature are 7.5 and 50oC,respectively.The protease activity is largely enhanced by Ca2 +,but highly inhibited by tetrasodium ethylenediaminetetraacetate(EDTA),a metal-chelator,suggesting that the enzyme is a metalloprotease.The Michaelis-Menten constan Km and Vmax value for casein substrate are 6.09 mg·ml -1and 21.32μg·min -1·ml -1, respectively.In vitro anthelmintic tests of the protease exhibit distinct lethal effects on the third stage larvae(L3)of Ascaris suum.Scanning electron microscopy and SDS-PAGE analysis indicates that the proteolysis of larvae proteins caused by this protease may relate to the anthelmintic activity of L.mylittae.

Variations of antioxidant enzyme activity and malondialdehyde content in nemertean Cephalothrix hongkongiensis after exposure to heavy metals

The antioxidant enzyme activity and malondialdehyde(MDA) content of Cephalothrix hongkongiensis were studied to assess variations in the biochemical/physiological parameters of nemerteans under heavy metal stress.Worms were exposed to copper,zinc and cadmium solutions at different concentrations,and the activity of three antioxidant enzymes,catalase(CAT),superoxide dismutase(SOD),and glutathione peroxidase(GPX),and MDA content were measured.The results show that the activity of each enzyme changed immediately after exposure to heavy metals.CAT was invariably inhibited throughout the experimental period,while the SOD activity was significantly elevated by exposure to Cu^(2+) for 48h,but then decreased.SOD was inhibited by Zn^(2+) during the first 12h of exposure,but activated when exposed for longer periods.Under Cd^(2+) stress,SOD activity decreased within 72h.GPX activity varied greatly,being significantly increased by both Cu^(2+) and Zn^(2+),but significantly inhibited by Cd^(2+) in the first 12-24h after exposure.MDA content increased on Cu^(2+) exposure,but normally decreased on Zn^(2+) exposure.MDA content followed an increase-decrease-increase pattern under Cd^(2+) stress.In conclusion,the antioxidant system of this nemertean is sensitive to heavy metals,and its CAT activity may be a potential biomarker for monitoring heavy metal levels in the environment.

Manila clam Venerupis philippinarum as a biomonitor to metal pollution

The Manila clam Venerupis philippinarum is a good biomonitor/bioindicator to marine metal pollution and is frequently used in aquatic toxicology. Two dominant pedigrees (white and zebra) of clam are distributed in the Bohai Sea; however, little attention has been paid to potential biological differences between these two pedigrees. In this study, we tested the sensitivity of both pedigrees to marine metal (cadmium and zinc) pollution biomonitoring and marine environmental toxicology. Results demonstrate significant biological differences in gills of white and zebra clams based on metabolic profiles and antioxidant enzyme activities. In addition, we found that hypotaurine, malonate and homarine were relatively high in white clam gills, while alanine, arginine, glutamate, succinate, 4-aminobutyrate, taurine and betaine were high in zebra clam gills. Zebra clam gills were also more sensitive to a mixture of Cd and Zn, as shown by antioxidant enzyme activities and metabolic profiles, but white clam gills could accumulate more Zn. Therefore, we suggest that the white pedigree can be used as a biomonitor to marine Zn pollution, whereas the zebra pedigree can be used for toxicology studies on Cd and Zn mixed pollution.

Comparison of Biochemical Properties of Recombinant and Native Phanerochaete flavido-alba Laccases

Laccases are blue copper oxidases （E.C. 1.10.3.2 benzenediol：oxygen oxidoreductase） that catalyze the one-electron oxidation of phenolics, aromatic amines and other electron-rich substrates with the concomitant reduction of 02 to H20. They are currently seen as highly interesting industrial enzymes because of their broad substrate specificity. The Phanerochaete flavido-alba laccase is expressed and secreted as a soluble active enzyme by Aspergillus niger （rLac-LPFA）. rLac-LPFA is easily purified to homogeneity. Metal ions like HgCI2, KC12, FeSO4 and MgSO4 at a concentration of 2 mM have inhibiting effect on recombinant and native laccase, whereas, CuSO4 and MnSO4 moderately increase both enzyme activities. Two potential inhibitors （sodium azide and EDTA） inhibited enzyme activity, whereas, urea and SDS have no effect on enzyme activity. The Km and V,,ax values for recombinant laccase are 0.65 mM and 300 U/mg respectively for 2,6-DMP as substrate.

Effect of warming Yang and removing blood stasis method on matrix metalloproteinases/tissue inhibitor metalloproteinases levels secreted by cultured endometrial cells from patients with endometriosis

OBJECTIVE:To investigate the effect of Chinese medicines using the warming Yang and removing blood stasis method on levels of matrix metalloproteinases(MMPs)/tissue inhibitor metalloproteinases(TIMPs) secreted by cultured endometrial cells from patients with endometriosis.METHODS:Ectopic and eutopic endometrial cells obtaind from 15 endometriosis patients were cultured in vitro,and divided randomly into five groups:high dose;moderate dose;low dose;nemestran;blank control.The three dose groups were treated with a decoction prepared according to the principle of warming Yang and removing blood stasis;nemestran and 0.9%NaCI were administered to the nemestran group and balnk control group,respectively.Eutopic endometrial cells obtaind from 10 hysteromyoma patients were cultured in vitro,as the normal control group,0.9%NaCI were administered to the normal control group.Cell culture supernatants were collected and levels of matrix metalloproteinase-1(MMP-1),matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9),tissue inhibitor metalloproteinase-1(TIMP-1) and tissue inhibitor metalloproteinase-2(TIMP-2) detected by enzyme-linked immuno sorbent assay(ELISA).RESULTS:Compared with the normal control group,levels of MMP-1,MMP-2,and MMP-9 in eutopic and ectopic endometrium cell supernatants in the blank control group were increased,whereas levels of TIMP-1 and TIMP-2 were decreased(P <0.05).Compared with the blank control group,levels of MMP-1 and MMP-2 in ectopic and eutopic endometrium cell supernatants cultured in low-dose,middle-dose,and high-dose groups were decreased,whereas levels of TIMP-1 and TIMP-2 were increased significantly(P < 0.05).CONCLUSION:The warming Yang and removing blood stasis method affects expression of MMPs andTIMPs.