Halomonas sp.BYS-1 was a moderately halophilic bacteria strain isolated from activated sludge,It could grow on MM with NaCl concentration from 0.1～2.6 mol/L and phenyl acetic acid as sole carbon souce. When BYS-1 grew in the media with different concentrations of NaCl, there was no obvious change in its intracellular Na+ contents , it accumulated K+, glutamic acid and betaine as osmoprotectants. Its intracellular contents of K+,glutamic acid and betaine increased by 1.9,2.4 and 13.6 times, respectively, when the concentration of NaCl increased from 0.1 mol/L to 2mol/L.

This study was conducted to investigate the physicochemical properties of Polyporus umbellatus sclerotial exudate. Morphological characteristics of the sclerotia and its exudate were observed during different stages of sclerotial formation. The pH of the exudate was detected at different time during cultivation. A phenol-sulfuric acid method was employed to determine the polysaccharide content of P. umbellatus sclerotial exudate during cultivating time. Additionally, the protein content was measured by means of BCA protein assay. Furthermore, CAT content was detected using ultraviolet absorption method. That the protein content of the exudate and CAT specific activity rose gradually during the passage of the cultivating time indicated a high level of oxidative stress during P. umbellatus sclerotial exudate formation. The results showed that the pH of the exudate increased gradually and then dropped down during sclerotial formation. That the pH of the exudate maintained the acidity state during the cultivation indirectly indicated that acidic environment would help sclerotial formation. The exudate produced gradually and was absorbed by the sclerotia itself.
Culture Media ; chemistry ; metabolism ; Fungal Proteins ; chemistry ; metabolism ; Fungi ; chemistry ; metabolism ; Hydrogen-Ion Concentration ; Medicine, Chinese Traditional ; methods ; Oxidative Stress ; Polyporus ; chemistry ; metabolism ; Polysaccharides ; chemistry ; metabolism

Acetamides ; pharmacology ; Animals ; Bridged-Ring Compounds ; poisoning ; Disease Models, Animal ; Female ; Male ; Rats ; Rats, Sprague-Dawley ; Troponin I ; blood

Humans ; Ischemic Postconditioning ; Myocardial Reperfusion Injury

HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular ; Escherichia coli ; metabolism ; HIV Integrase ; metabolism ; HIV-1 ; physiology ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Protein Binding ; Virus Replication

AIM: To obtain nerve growth factor subunit ?(NGF ?) gene from Chinese fetal hippocampus tissue, prove its sequence, clone the mature peptide sequence, and make it express in E.coli .METHODS: Total RNA was extracted, amplified by RT-PCR method. Its 650 bp DNA sequence was inserted into PCR Ⅱ vector. PCR/NGF ? vector was used as the template to amplfy the C-terminate mature peptide sequence, then subcloned it into PG5 vector. The recombinant was transferred into E.coli BL21. BL21 was cultured and induced by IPTG. The activity of the expressed product was measured after purified and refolded.RESULTS: A complete cDNA sequence was determined as 1 047 nucleotides. The cloned 636 bp encoding 212 amino acids was proved homological to Genbank by sequence analysis. The expressed mature peptide showed an clear band of the prospected 14 kD by SDS-PAGE electrophoresis, and was testified by Western blot. The expression level was about 10% of the total cell lysate. CONCLUSION: The chinese NGF ? gene was homological to the foreigners'. The recombinant NGF ? was efficiently expressed in E.Coli and the recombinant protein has high immunological activities.

Objective To find the relationship between mutation of gamma 2 subunit of the gamma-aminobatyric acid type A receptor(GABRG2) and generalized epilepsy with febrile seizure plus(GEFS+).Methods Probands of 10 families with GEFS+ were selected,the GABRG2 gene were sequenced.Results We found a single nucleotide polymorphism site,and did not find the reported mutations.Conclusion GABRG2 mutation is not common in Hans of northern China.

Objective To observe the changes of femur biomechanical properties in fluorosis rats. Methods Fifty Wistar rats of thirty-day old, weighing 90-100 g, were randomly divided according to body mass into fluorosis and control group of 25 each. Fluorosis group drank tap water containing 100 mg/L of fluoride, the control group drank tap water. The rats were observed of dental growing status and killed after feeding 6 months. Their femurs underwent tensile strength, impact, shear, bending experiments. Results The deteetable rate of dental fluorosis was significantly higher in fluorosis group[92%(23/25)] than control group[0(0/25),X2=38.97, P<0.01]. Biomeehanical data in fluorosis group(225.67±11.81,1.94±0.15,76.62±6.10,39.96.3±3.90) were lower than those of the control group(244.70±13.38,2.39±0.19,87.72±7.05,45.75±3.75) in experiments of tensile strength (MPa), impact toughness (J/mm2), shear and bending strength (MPa). The difference was statistically significant(t=3.372,5.879,3.756 and 3383, respectively, P<0.01) between the two groups. Conclusion Fluorides affected bone metabolism in rats, femur biomechanieal properties ehanged in fluorosis rats.