8 trace elements including Cu, P, Zn, Mg, Fe, Ca, Mn, and Ba were determined in 60 volunteers and 41 patients with endemic fluorosis. The results showed that in the patient's group serum Cu was substantially higher and serum Zn as well as Fe were lower while hair Cu and Mg were both significantly elevated as compared with controls.Based on the results mentioned above, it may be suggested that hair could be used as an alternative source for clinical measurement of trace elements because of its easy availability. The significance of the changes in the 8 trace elements was also discussed, but the pathological mechanism has been unknown yet.

Objective To construct a bait plasmid in bacterial two-hybrid system.Methods A cDNA fragment encoding for rat AMP-activated protein kinase ?2(AMPK?2) was amplified by PCR and inserted into bacterial expression vector pBT.After confirmation with restricted endonuclease digestion and sequence analysis,bacterial reporter strain XL-1 Blue MRF' was transformed with pBT-AMPK?2 plasmid and the expression of the recombinant bait fusion protein was detected.To test whether the bait fusion protein had the capability of self-activation,the XL-1 Blue MRF' cells were cotransformed with the pBT-AMPK?2 plasmid and empty pTRG vector,and screened on 3-amino-1,2,4-triazole(3-AT) Selective Screening Medium plates.Results Restriction digestion and sequence analysis revealed that the AMPK?2 code sequence was correctly inserted into pBT with a right reading frame.pBT-AMPK?2 expressed ?cI/AMPK?2 fusion protein.Colonies were obtained on no 3-AT Nonselective Screening Medium plates when XL-1 Blue MRF' cells were cotransformed with recombinant pBT-AMPK?2 and empty pTRG vector,while none grew on 3-AT plates,indicating that the recombinant plasmid pBT-AMPK?2 expressed AMPK?2/?cI fusion protein correctly,and was incapable of activation of the reporter cassette in the absence of an interaction partner.Conclusion The recombinant plasmid pBT-AMPK?2 could be used as "bait plasmid" to screen cDNA library.

AIM: To explore the effect of acute and chronic hypoxia on the chemotaxis of polymorphonuclear leukocytes(PMNs) of rats.METHODS: Serum and PMNs were separated from rats under normoxia, acute hypoxia and chronic hypoxia condition,and cell aspiration technology system was used to determine the chemotactic migration of pseudopod of PMNs induced by serum as chemotactic factor.RESULTS: Induced by autoserum,the length of pseudopod of PMNs migrated into micropipette [(1.200?0.178)?m,(1.094?0.164)?m ]under acute hypoxic and chronic hypoxic condition was significantly longer than the corresponding value for control[(0.914?0.156)?m,P

With the advent of post-genomic era, identification of protein-protein interaction has become another hot spot of protein research and promoted the invention, development and complement of relative techniques. Major large-scale high-throughput methods, such as two-hybrid system, bacteriophage display, mass spectrometry, protein microchips and bioinformatics have provided a global view of protein-protein interaction, and are hopeful to play an important role in proteomics. Each method may have its specific strengths and weaknesses as well as differences in coverage, to some extent, the data based on these methods can complete each other. Here, recent progress of these large-scale high-throughput methods and their applications for studying protein-protein interaction are reviewed.

AIM: To determine the effect of agmatine on the proliferation of pulmonary artery smooth muscle cells (PASMCs) induced by hypoxia. METHODS: Primary culture of rat PASMCs was prepared from adult male Wistar rat pulmonary artery by the method of tissue block anchorage. PASMCs were divided into two groups: control group and agmatine-treated group. The activity of LDH in the medium was measured by chromatometry. [3H]-TdR incorporation was measured by liquid scintillation counting. The cell cycle was measured by flow cytometry, and the PCNA content was measured by image analysis. RESULTS: Agmatine did not exert significant effect on the activity of LDH in the medium, but significantly decreased the [3H]-TdR incorporation and PCNA content of PASMCs. Agmatine significantly decreased the cell ratio of G_2/M phase and increased the cell ratio of G_0/G_1 phase. With the increment of the concentration of agmatine, [3H]-TdR incorporation was significantly decreased. CONCLUSION: Agmatine has no significant cytotoxic effect on PASMCs and agmatine dose-dependently inhibits the proliferation of rat PASMCs induced by hypoxia.

Estrogen has many physiological functions. It can prevent atherosclerosis, coronary heart disease and osteoporosis etc. Phytoestrogen is a kind of natural compounds in plants that is structurally similar to estrogen and has estrogen-like effects. The studies on the effects of estrogen and phytoestrogen on the hypoxia at present is limited and its mechanism is being researched.

Objective To investigate the roles of hypoxia regulated genes/protein in chemical hypoxia preconditioning in differentiated SH-SY5Y cells.Methods Differentiated SH-SY5Y cells were randomly divided into control group,chemical hypoxic preconditioning group(50 ?mol/L CoCl2 preconditioning for 3 h,normal culture for 1 h,then in 2%O2 for 28 h) and hypoxia group(in 2%O2 for 28 h).RT-PCR was used to examine the mRNA expressions of VEGF,GLUT-1,EPO,LDH-A.Further evaluation of the potential neuroprotective effect of VEGF was done by investigating the effect of recombinant human VEGF on subsequent hypoxia injury.Results The mRNA levels of GLUT-1,EPO increased in the preconditioning group as compared with those in the hypoxia group(P

Objective:To determine the content of magnolol and honokiol in KAIXIONG SHUNQI CAPSULE by HPLC.Methods:Separation was achieved on a Nova Pak C 18 column. The mobile phase was metanol water(78∶22), the wavelength of the UV detector 294nm.Results:Linear range of megnolol and honokiol were 0.08?g 0.4 ?g and 0.052?g 0.26?g, respectively. The average recovery was 100.22% ( RSD =1.21%).Conclusion: The method was accurate and reliable.

Objective To screen the proteins interacting with AMP-activated protein kinase ?2(AMPK?2)in rat brain cDNA library by bacterial two-hybrid system,and to investigate the biological role and the regulatory mechanism of AMPK?2 in brain.Methods The recombinant pBT-AMPK?2 was used as the bait to screen a rat fetal brain cDNA library by bacterial two-hybrid system.The plasmids of positive colonies were extracted and analyzed by DNA sequencing and BLAST search in GenBank.Results Seven proteins binding to AMPK?2 were screened out,including polyubiquitin,small heat shock protein 8,phosphofructokinase,cytochrome C oxidase subunit Ⅰ(COXⅠ),HLA-B-associated transcript 3(BAT3)isoform 1,protein tyrosine phosphatase receptor type D(Ptprd)and islet-brain 1(IB1).Conclusion Polyubiquitin,HSP8,phosphofructokinase,COXⅠ,BAT3 isoform 1,Ptprd and IB1 can interact with AMPK?2 in brain.

Objective To investigate the effects of hypoxia on the rheologic properties of leukocytes in microcirculation of rats, and its relationship with the expressive changes of CD18 of neutrophils. Methods Hypobaric chamber was used to simulate the plateau condition for the establishment of animal hypoxic model. Adult Wistar rats were randomized in three groups: control group, short-term hypoxia group (Hypobaric chamber simulated 5000m altitude for 3days) and long-term hypoxia group (Hypobaric chamber simulated 5000m altitude for 30 days). Intravital microscopy and computer image analysis system were used to evaluate the rheologic properties of leuckcytes in mesentery microcirculation. Neutrophils of the three experimental groups were purified and the expression of CD18 was determined by flow cytometer. Results Compared with control group, the rheologic properties of leukocytes were changed obviously at both short-term hypoxia and long-term hypoxia groups, including the number of leukocytes rolling and adherenced on endothelium increased, TLECT (total leukocyte-endothelium contact time) obviously extended (P