Fibrolase is a non-hemorrhagic zinc metalloproteinase isolated from southern copperhead snake venom (Agkistrodon contortrix contortrix) and is capable of degrading fibrin clots resulting from purified fibrinogen or from blood plasma. Alfimeprase, a truncated form of fibrolase, as a clinical agent was successfully completed PhaseII clinical trials.The cDNA of alfimeprase was amplified by recursive PCR, digested with BamHI and HindII, and cloned into pET43.1a, pMALp2X and pMALc2X vectors to generate fusions with NusA, MBP and sMBP(with signal peptide), respectively. Nus/alfimeprase was expressed in soluble form by co-expressing with chaperone FkpA and inducing with1mmol/L IPTG. The fusion protein accounted for about 25 % of total protein following cell lysis. Alfimeprase was successfully purifiesd by Ni-NTA affinity chromatography and cleaved by enterokinase. The results demonstrate the fibrinolytic activity of recombinant alfimeprase using fibrin plate assays and fibrinogen hydrolysis.

Background Glaucoma is one of the leading causes of blindness worldwide;while the different types of glaucoma is vary from different region. Objective Present study was to survey the prevalence and types of glaucoma among peasants of Uigur adults in Kuche county and offer the basis for the prevent and treatment of glaucoma in Uigur nationanlity. Methods 4191 Uigur peasants aged 40 years or above were collected in Kuche county for the survey of prevalence and types of glaucoma by randomized cluster sampling in March and April of 2009. The subjects were grouped into 40-,50-,60- and ≥70 years groups according to the distribution of age. The disease history of glaucoma, regular eye examination, funds examination and measurement of the anterior chamber depth, gonioscope were performed in the all subjects. Darkroom prone test and mydriasis test were carried out in suspicious glaucomous patients. The depth of periphery anterior chamber was assessed based on van Herick' s criteria, and the width of chamber angle was graded based on Scheie' s method. The standardized training was performed. This survey approved by Xinjiang Medical Ethics Committee, all subjects signed the informed consent before the examination. Results 4191 of 4873 subjects finished all the examinations with the response rate 86%. All the subjects showed a good compliance. The prevalence of glaucoma was 3. 79% , and the prevalence of primary angle-closed glaucoma(PACG) .primary angle-open glaucoma (PAOG) and secondary glaucoma was 2.22% ,0.26% and 1. 31% respectively, showing a significant difference among the different types of glaucoma( P<0. 05). The prevalence of glaucoma was elevated with aging (χ2 - 116. 69 ,P<0. 05) and presented a high rate in male subjects compared with female ones(χ2 = 7. 34, P<0. 05 ). Bilateral blindness was found in 19.75% glaucoma peasants, in which 25.3% glaucoma peasants received anti-glaucoma surgery. The distribution of visual acuity of patients was of significant difference among different age groups(χ2 = 37. 69 ,P<0. 05 ) . Conclusions The prevalence of the glaucoma among Uigur peasants in Kuche county was higher than most area no matter inland or overseas. PACG still is the common type in those people.

Objective To investigate the levels of serum vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) in patients with non-small cell lung cancer(NSCLC) and relationships with c1inicopatho1ogica1 characteristics and their clinical significance. Methods The concentrations of serum VEGF and bFGF were detected by enzyme-linked immunosorbent assay(ELISA) in 40 patients with NSCLC before and after chemotherapy. Results The level of serum VEGF in patients with Ⅳ stage NSCLC was significantly higher than that of Ⅲ stage(P

OBJECTIVETo study the effect of interferon-gamma (IFN-gamma) on the proliferation of mesangial cells (MsC) and transforming growth factor (TGF)-beta/Smad signal pathway, the mRNA and protein expression of matrix metalloproteinase-2 (MMP-2) and tissue inhibitors of matrix metalloproteinase-2 (TIMP-2), and to provide an experimental basis for IFN-gamma treatment of renal fibrosis.

METHODSCultured MsC were treated with IFN-gamma at different concentrations and the proliferation of MsC was examined by MTT. Protein and RNA samples were extracted from MsC at 0, 0.5, 1, 2, 4, 6, 12, 24 h after treated by 100 IU/ml IFN-gamma. The mRNA and protein expression of Smad3, Smad7, MMP-2 and TIMP-2 were analyzed by real-time RT-PCR and Western blot, respectively.

RESULTSThe expression of Smad7 mRNA and protein were promptly elevated at 0.5 hour after the IFN-gamma treatment and lasted for 6 hours, but the proliferation of MsC was not altered. The elevated expression of Smad3, MMP2 mRNA and proteins persisted after 6 hours, whereas the expression of TIMP-2 mRNA and protein decreased.

CONCLUSIONThe therapeutic effect of IFN-gamma of renal fibrosis may be mediated by TGF-beta/smads signal pathway through up-regulation of MMP-2 expression, coupled with down-regulation of TIMP-2 expression.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Interferon-gamma ; pharmacology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Mesangial Cells ; cytology ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Recombinant Proteins ; Signal Transduction ; Smad3 Protein ; genetics ; metabolism ; Smad7 Protein ; genetics ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; metabolism ; Transforming Growth Factor beta ; metabolism

OBJECTIVETo investigate the changing of T4 5'-and 5-deiodinase within rat brain under various iodin-nutritional states.

METHODSAnimal model of iodine-deficiency rat was performed and the rats were divided into 4 groups by the intake of iodine-nutrition, and then killed at an age of 20 days. The thyroid hormones level in serum was measured by ELISA and the activity of T(4) 5'-and 5-deiodinase within brain was analyzed.

RESULTSIn less-iodine (LI) group,TT4 and FT4 were accounting for 3.5% of the neutral-iodine (NI) group's, and FT3 was 174.0% of NI group's (P < 0.05). In NI group,TT4 and FT4 were 114.5% and 127.7% of NI group's (P < 0.05). In high-iodine (HI) group, TT4 and FT4 were 61.86% and 62.0% of NI group's, and FT3 was 184.9% of NI group's (P < 0.05). In LI group, the activity of T4 5'-deiodinase tissue of per gram (1.95 +/- 0.32) ngT3.microgT4(-1).h was significantly higher than that of NI group (P < 0.05), and the activity of 5-deiodinase (1.38 +/- 0.21) ngrT3.microg T4(-1).h(-1) is significantly less than that of NI group (1.59 +/- 0.23) (P < 0.05). In HI group the activity of T4 5'-and 5-deiodinase tissue of per gram (1.12 +/- 0.19 and 1.73 +/- 0.36) ngrT3.microgT4(-1).h(-1)was significantly less than that of NI group (P < 0.05).

CONCLUSIONThe activity of T4 5'-deiodinase in iodine deficiency heightens and that in iodine excess is debased, the activity of T4 5-deiodinase in iodine deficiency and in iodine excess is debased.

Animals ; Brain ; enzymology ; Female ; Iodide Peroxidase ; metabolism ; Iodine ; administration & dosage ; deficiency ; Male ; Rats ; Rats, Wistar ; Thyroxine ; blood ; Triiodothyronine ; blood

OBJECTIVETo evaluate the efficacy and safety of using Jiangzhi Tongluo Soft Capsule (JTSC) combined with Atorvastatin Calcium Tablet (ACT) or ACT alone in treatment of combined hyperlipidemia.

METHODSA randomized, double blinded, parallel control, and multi-center clinical research design was adopted. Totally 138 combined hyperlipidemia patients were randomly assigned to the combined treatment group (A) and the atorvastatin treatment group (B) by random digit table, 69 in each group. All patients took ACT 20 mg per day. Patients in the A group took JTSC 100 mg each time, 3 times per day. Those in the B group took JTSC simulated agent, 100 mg each time, 3 times per day. The treatment period for all was 8 weeks. Serum levels of triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) were observed before treatment, at week 4 and 8 after treatment; and safety was assessed as well.

RESULTSAt week 4 and 8 after treatment serum TG decreased by 26.69% and 33.29% respectively in the A group (both P < 0.01), while it was decreased by 25.7% and 22.98% respectively in the B group (both P < 0.01). At week 8 decreased serum TG was obviously higher in the A group than in the B group (P < 0.05). Compared with before treatment, serum levels of LDL-C and TC levels decreased significantly in the two groups (all P < 0.01). There was no statistical difference in the drop-out value and the drop-out rate of serum LDL-C and TC levels (P > 0.05). At week 8 the serum HDL-C level showed an increasing tendency in the two groups. No obvious increase in peptase or creatase occurred in the two groups after treatment.

CONCLUSIONJTSC combined with ACT could lower the serum TG level of combined hyperlipidemia patients with safety.

Adult ; Atorvastatin Calcium ; Double-Blind Method ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Heptanoic Acids ; therapeutic use ; Humans ; Hyperlipidemias ; drug therapy ; Male ; Middle Aged ; Pyrroles ; therapeutic use ; Treatment Outcome ; Triglycerides ; blood

This study was aimed to investigate the effect of recombinant human granulocyte colony-stimulating factor(G-CSF) on murine thymocyte emigration and cell cycle alteration after a sublethal dose of gamma-irradiation. Female BALB/c mice were given 6.0 Gy γ-ray total body irradiation and then randomly divided into G-CSF and control groups. Mice in the G-CSF group were injected recombinant human G-CSF 100 µg/(kg·d) subcutaneously once daily for 14 consecutive days and mice in the control group were given the same volume of phosphate buffered solution. Thymocyte cycle alteration and the proportion of apoptosis cells were detected by flow cytometry within 72 hours after irradiation. Real-time PCR was used for detection and quantitation of murine T cell receptor rearrangement excision circles (sjTREC) of the thymic cells at 30 and 60 day after the irradiation. The results showed that at 6 hour after irradiation G-CSF could significantly increase the thymic cells in G(0)/G(1) phase, G-CSF vs control: (82.0 ± 5.0)% vs (75.9 ± 2.8)% (p < 0.05), and decrease the thymic cells in S phase, G-CSF vs control: (10.2 ± 4.8)% vs (15.7 ± 2.3)% (p < 0.05), but G-CSF seemed have no evident effects on the percentage of thymic cells in G(2)/M phase. G-CSF could also protect thymocytes from apoptosis at 6 hour and 12 hour after irradiation the percentages of apoptosis cells in G-CSF group were (11.5 ± 2.4)% and (15.5 ± 3.3)%, respectively, which were significantly lower than that of the control group (16.5 ± 2.2)% and (22.6 ± 0.7)%, respectively (p < 0.05). The sjTREC copy amount was conspicuously higher in G-CSF group than that in the control at 30 day after irradiation (p < 0.01), but the preponderance disappeared 60 days later. It is concluded that G-CSF has a positive effect on the thymic cell cycle alteration to protect thymocytes from apoptosis and enhance the recent thymocyte emigration, which may contribute to the central immune reconstitution after irradiation.
Animals ; Cell Cycle ; drug effects ; radiation effects ; Female ; Gamma Rays ; adverse effects ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; pharmacology ; Thymocytes ; drug effects ; radiation effects

OBJECTIVETo examine the effect of multiple micronutrients supplementation on anti-oxidative activity and decreasing oxidized DNA damage of lymphocytes in Chinese children.

METHODS82 healthy children in a rural areas, aged 9-11 years, were selected and randomized allocated into group receiving supplements and control group with each of them 41. 24-hour dietary recall was used to collect data on daily nutrient intakes of the research subjects. The subjects in the supplement group were given vitamin A (VA) 600 microg, beta-carotene (beta-C) 1.0 mg, vitamin E (VE) 100 mg, vitamin C (VC) 300 mg and Na2SeO3(Se) 200 microg in a tablet on daily base while those in the control group took a same-sized color placebo tablet. The trial lasted 8 weeks. 5 ml blood samples from each subject were taken during 7 to 9 o'clock in the morning. DNA damage of lymphocytes and levels of plasma VA, VE, VC, beta-C, Se, malondialdehyde (MDA), activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were then analyzed twice before and after the 8-week of trial.

RESULTSThe low intakes of VA, VC and Se only accounted for 50.6%, 65.6% and 67.3% of their recommended nutrient intake (RNI) respectively. After the trial, levels of plasma beta-C, VA, VE, VC and Se in the supplemented group increased by 13.4%, 32.8%, 11.5%, 46.9% and 24.6% respectively, compared with the control group, indicating that nutritional status regarding antioxidant nutrients had largely been improved. GSH-Px activity had a significant increase in the supplement group than before the supplement and in the control group (P < 0.01). GSH-Px before the trial (the 100.4 U/ml) also showed significant increase after the trial (161.7 U/ml) (P < 0.01). However, the values of SOD and MDA significantly decreased after the trial. Analysis of DNA damage indicated that there was no significant difference in the intrinsic damage of DNA (P > 0.05). Significant decreases of oxidized DNA damage induced by 5 micromol/L, 10 micromol/L and 25 micromol/L H2O2 were found more in peripheral lymphocytes of the supplemented group, than in pre-supplement and the control group after the trial (P < 0.01).

CONCLUSIONSupplementation of multiple micronutrients could effectively increase the levels of beta-C, VA, VE, VC and Se in plasma, and GSH-Px activity. In the meantime, MDA and oxidized DNA damage induced by a low level H2O2 decreased significantly after the trial. The reason accounted for the decrease of SOD activity after the trial needs to be further studied.

Antioxidants ; administration & dosage ; Ascorbic Acid ; administration & dosage ; Child ; China ; DNA Damage ; drug effects ; Dietary Supplements ; Female ; Humans ; Lymphocytes ; metabolism ; Male ; Malondialdehyde ; blood ; Nutrition Surveys ; Oxidative Stress ; drug effects ; genetics ; Rural Health ; Selenium ; administration & dosage ; Superoxide Dismutase ; blood ; Vitamin A ; administration & dosage ; Vitamin E ; administration & dosage ; Vitamins ; administration & dosage

An alkaline catalase has been purified and characterized from a slightly halophilic and alkaliphilic bacterium Bacillus sp. F26. The purification was performed with a four step procedure consisting of ammonium sulfate precipitation, ion exchange, gel filtration and hydrophobic interaction chromatography, and finally achieved a 58.5-fold-purifying over the crude extract. The purified catalase was composed of two identical subunits with a native molecular mass of 140 kD. The native enzyme showed the typical Soret band appearing at 408 nm. The pyridine hemochrome spectrum indicated the presence of protoheme IX as the prosthetic group. The apparent Km value for enzyme activity on H2O2 was calculated to be 32.5 mmol/L. The activity of this catalase was not reduced by dithionite but was strongly inhibited by cyanide, azide, and 3-amino-1,2,4-triazole (the specific inhibitor of monofunctional catalase). No peroxidase activity of this enzyme was detected when using o-dianisidine, diaminobenzidine (DAB) and p-phenylenediamine as electron donor. Moreover, the N-terminal sequence of this catalase exhibited substantial similarity to the monofunctional catalase subgroup rather than catalase-peroxidase or Mn-catalase one. Therefore, we characterize the purified catalase as a monofunctional catalase. Besides, this monofunctional catalase was thermosensitive and its activity exhibited pH-independent over pH 5-9 but showed a sharp maximum at pH 11. An activity half-life of approximately 49 h was measured when the enzyme was incubated at 20 degrees C and pH 11. To our knowledge, pH 11 is the most alkaline condition for optimum catalysis and enzyme stability among the catalases reported up to now. Furthermore, this monofunctional catalase also showed excellent halo-alkali-stability with a half-life of approximately 90 h at 0.5 mol/L NaCl and pH 10.5. On the other hand, so far as we know, the characterized catalase is the first dimeric monofunctional catalase from alkaliphiles and is also the first monofunctional catalase derived from a natural soda lake, which could partially reflect the oxidative stress response in the corresponding environment.
Bacillus ; enzymology ; Bacterial Proteins ; chemistry ; isolation & purification ; Catalase ; chemistry ; isolation & purification ; Enzyme Stability ; Hydrogen-Ion Concentration ; Temperature

OBJECTIVETo compare and explore the physiological and biochemical index of virus-free seedling and common seedling of Chrysanthemum morifolium.

METHODLeaves of virus-free seedling and common seedling were cut down and the contents of chlorophyll a and b, MDA, and the activity of SOD, POD, CAT, and the net photosynthetic rate (Pn) were measured.

RESULT AND CONCLUSIONThe contents of chlorophyll and Pn of virus-free seedling were higher than those of common seedling. But were measured, the contents of MDA and the activity of SOD, POD, CAT related to the physiological resistance were lower than the latter. These could explain the phenomenon of virus-free seedling grow better than the common seedling under the same condition.

Catalase ; analysis ; Chlorophyll ; analysis ; Chrysanthemum ; metabolism ; physiology ; virology ; Malondialdehyde ; analysis ; Peroxidase ; analysis ; Photosynthesis ; Plant Leaves ; chemistry ; physiology ; Plant Viruses ; pathogenicity ; Plants, Medicinal ; metabolism ; physiology ; virology ; Seedlings ; chemistry ; physiology ; Superoxide Dismutase ; analysis