Objective To concentrate coagulation Factor Ⅺ(FⅪ)from human plasma.Methods FⅪwas isolated and purified from human plasma via two-step chromatography,including CM-Sepharose fast flow ion exchange and Heparin CL-6B affinity chromatography.Results The recovery rate of FⅪ was(21.02?5.04)%,the specific clotting activity of purified FⅪ was(17.59?1.96) U/mg,and the purification factor was(1162.29?129.64) fold(n=7).Conclusion The two-step chromatography is effective in concentrating FⅨ.

OBJECTIVETo study the effect of Bushen Tongdu Capsule (BTC) on RANK/RANKL/ OPG pathway of collagen induced arthritis (CIA) rats, thereby laying theoretic evidence for treating rheumatic arthritis (RA) by Chinese medicine.

METHODSRA model was induced by CIA. Totally 42 rats were randomly divided into six groups, i.e., the normal control group, the model group, the low dose BTC (BSL) group, the medium dose BTC (BSM) group, the high dose BTC (BSH) group, and the Tripterygium Glycosides (TG) group, 7 in each group. BTC at the daily dose of 120, 240, and 480 mg/kg was given by gastrogavage to rats in the BSL, BSM, and BSH group respectively from the 13th day of modeling. TG at the daily dose of 24 mg/kg was given by gastrogavage to rats in the TG group. All medication was given once daily, 2 mL each time. Two mL normal saline was administered to rats in the normal control group and the model group. All medication lasted for 18 days. Samples were taken at day 31. The TRAP section of the ankle joint was fixed in 10% formalin for TRAP stain. Serum levels of osteoprotegerin (OPG), receptor activator of nuclear factor-κB ligand (RANKL), and macrophage colony-stimulating factor (M-CSF) were detected using ELISA.

RESULTSCompared with the normal control group, positive reactions of pathological ankle joint section, inflammation, and osteoclasia degree were significantly improved in the model group, serum levels of RANKL and M-CSF were up-regulated, levels of OPG and OPG/RANKL were significantly lowered (all P < 0.01). Compared with the model group, positive reactions of pathological ankle joint section, inflammation, and osteoclasia degree also significantly decreased in the BSH group and the TG group (all P < 0.01). RANKL and M-CSF were significantly down-regulated in each medicated group, while levels of OPG and OPG/RANKL were significantly up-regulated (all P < 0.01). Compared with the TG group, M-CSF was lower, but levels of OPG and OPG/RANKL were significantly up-regulated in the normal control group (all P < 0.01). RANKL and M-CSF were significantly up-regulated, while levels of OPG and OPG/RANKL were significantly down-regulated in the model group and each BS group (all P < 0.01).

CONCLUSIONBTC could relieve bone damage of CIA rats possibly through regulating and controlling osteoclasts.

Animals ; Arthritis, Experimental ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; Macrophage Colony-Stimulating Factor ; metabolism ; Osteoclasts ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Rats ; Receptor Activator of Nuclear Factor-kappa B ; metabolism ; Tripterygium

Idiopathic thrombocytopenia purpura (ITP) is an autoimmune disease which is characterized by destruction of platelets by macrophages in the reticuloendothelial system. Recent studies suggest that ITP is related to the abnormal activation and apoptosis of T/B cells which lead to failure of immune tolerance. Now it is becoming clear that costimulatory signals are required for full T/B cell activation and assumed to modulate T/B cells responses as well as other aspects of the immune system. This review focuses on the role and state-of-the-art advancements of costimulatory signals in ITP.
Antigens, CD ; immunology ; Antigens, Differentiation ; immunology ; Antigens, Differentiation, T-Lymphocyte ; immunology ; B-Lymphocytes ; immunology ; CD28 Antigens ; immunology ; CTLA-4 Antigen ; Humans ; Immune Tolerance ; Inducible T-Cell Co-Stimulator Protein ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; Receptors, Tumor Necrosis Factor ; immunology ; Signal Transduction ; physiology ; T-Lymphocytes ; immunology ; physiology

Objective To investigate the effects of chronic high glucose on α-cells glucagon releasing in relation to insulin resistance induced by high glucose. Methods TC1-6 cells, an α-cell line, were incubated separately in DMEM containing high (25.0 mmol/L), medium (11.1 mmol/L) and low (5.5 mmol/L) concentrations of glucose for 1 to 5 days. The secretion and gene expression of glueagon were measured. When TC1-6 cells had been cultured for 5 days, three different concentrations of insulin were added for 6 h and then glucagon secretion was detected. Western blot was used for 1 and 3 days to confirm the effect of high glucose on phosphorylation of Akt in TC1-6 cells. Results (1) Exposure of TC1-6 cells to 11.1 and 25.0 mmol/L glucose resulted in a slight increase of glucagon secretion compared with those incubated with 5.5 mmol/L. However, after 5 days in media containing 25.0 mmol/L glucose, glucagan secretion was significantly increased as compared to cells treated with low glucose [(136.80±10.94 vs 78.62±4.72 ) ng/106 cells, P<0.05]; moreover, in TC1-6 cell cultured with high glucose glucagon mRNA expression was increased significantly. (2) 10-7 mol/L insulin reduced significantly glucagon secretion of TC1-6 ceils exposed to low glucose [(21.59±1.30 vs 55.12±3.86) ng/106 cells], but just scarcely inhibited glucagon secretion of cells incubated with high glucose [(106.58±8.53 vs 117.18±10.55) ng/106 cells]. When insulin concentration was increased to 10-5 mol/L, glucagon secretion of TC1-6 cells in high glucose was also reduced [(46.55±3.72 vs 118.61±10.68 )ng/106 cells]. (3) After treated with 10-5 mol/L insulin for 2h, the levels of Akt phosphorylation in both groups of TC1-6 cells were increased by 180% and 70%, while the level in high glucose group was significantly lower than that in low glucose group. In the presence of phosphoinositide 3 kinase inhibitor, the levels of Akt phosphorylation were both lowered, but the inhibition in low glucose group was more significant than in high glucose group. Conclusion High glucose induces hypersecretion of glucagon, which may be due to the a-cell insulin resistance.

This paper proposes a one-way micro valve with a simple structure and a simulation design for the engineering capsule. We have now got its design parameter selection method and its mechanic characteristic from experiments.
Capsules ; Computer Simulation ; Drug Delivery Systems ; methods ; Equipment Design ; Infusion Pumps, Implantable ; Technology, Pharmaceutical ; instrumentation ; methods

Objective To establish the enzyme-linked immunosorbent assay (ELISA) methods for the quantitative determination of IgG antibodies against diphtheria (DT) and tetanus (TT).MethodsPurified diphtheria toxiod and tetanus toxoid were respectively used as the coating antigens,the human-derived serum antibody standard substance of DT and TT served as the standard substance.The dose-response curves of the tested samples and standard substance were fitted.Then the two quantitative ELISA methods for determining the antibody to DT (Anti-DT) and antibody to TT (Anti-TT) were established with the parallel lines method.Then the methodological verification and application study were conducted.Results The validation results of the two quantitative ELISA measurement methods were in accordance with the regulations.The quantity limit of ELISA method for quantitative detection of Anti-DT demonstrated to be 0.084 mIU/mL,its average recovery rate was 97.6%.The intra-assay coefficient of variation(CV) and inter-assay CV of this Anti-DT assay were ≤ 3.40% and ≤5.05%,respectively.The quantity limit of ELISA method for quantitative detection of Anti-TT demonstrated to be 0.175 mIU/mL,its average recovery rate was 97.5%.The intra-assay CV and inter-assay CV of this Anti-TT assay were ≤ 2.42% and ≤5.58%,respectively.These two methods were applied for the immunogenicity evaluation after infantile basic immunization by diphtheria and tetanus vaccines.Conclusion The two established quantitative ELISA methods demonstrate high accuracy and good reproducibility,which are suitable for the ordinary laboratory to carry out the work and can be used in the serological effect evaluation after diphtheria and tetanus vaccine immunization and epidemiological study of diphtheria and tetanus disease.

Objective To knowthe effect of selenium on expression ofnitric oxide synthase(eNOS) and inducible nitric oxide synthase(iNOS) induced by fluoride in ECV-304(human umbilical vein endothelial cells).Methods The cells,ECV-304,were divided into two groups and treated as follows,one group was treated with sodium fluoride at different doses(0,400,1 000 and 2 000 ?mol/L),the other was treated with sodium fluoride at different doses(0,400,1 000 and 2 000 ?mol/L)and added sodiumselenite(100 nmol/L) respectively.24 h of treatment,The activity ofeNOS and iNOS were measured by NO and NOS assay kits and the expression of eNOS mRNA and iNOS mRNA was analyzed by RT-PCR.Results The activity of eNOS and the expression of eNOS mRNA were down-regulated by fluoride with a dose dependent manner,while the activity of iNOS and the expression of iNOS mRNA were up-regulated.Selenium showed a significant antagonism to low level fluoride treated groups.Conclusion Seleniumcan antagonize the effect offluoride on the expression ofiNOSand eNOS in the vein endothelial cells when the fluoride exposure level is not toohigh.

In this paper, five different methods were carried out for DNA extraction directly from soil. The result shows that all five methods could generate DNA with more than 15 kb in size. They were subsequently used as templates for PCR amplification with success, using primers of the bacterial 16S rRNA gene and Shiva-1 gene encoding an antibacterial peptide. However, method 5 is more suitable for DNA extraction directly from a small amount of soil sample as it produced a good yield of DNA in high integrity with reliable reproducibility.

Objective To study the effect of different concentrations of fluoride on cultured human umbilical vein vascular endothelial cells(HUVEC). Methods Different doses of sodium fluoride (NaF) were added to HUVEC culture medium, fluoride concentrations were 0(control), 100,400,700,1000,2000 μmol/L, respectively,6 re-set hole in each group. After continuous culture for 48 h, cells and culture medium were collected. Cell morphology was studied by Wright-Giemsa staining; cells apoptosis was determined by acridine orange fluorescence staining; cell activity was measured by methyl thiazolyl tetrazolium (MTT) assay; superoxide dismutase (SOD),glutathione peroxidase(GSH-Px) activity, malonaldehyde(MDA) content, induced nitricoxide synthase(iNOS), and endothelia nitricoxide synthase(eNOS) activity in cell culture medium were determined by spectrophotometry; cell iNOS mRNA and eNOS mRNA expression were detected by RT-PCR; intercellular adhesion molecule-1 (ICAM-1)and vascular cell adhesion molecule-1 (VCAM-1) levels were detected by double antibody sandwich ELISA method.Results With increased dose of fluoride, HUVEC cells decreased, the structure changed. In 400 - 2000 μmol/L group, the SOD activity[(6.627 ± 0.213), (6.668 ± 0.152), (5.935 ± 0.122), (4.755 ± 0.182)kU/L] was lower than those of the control group[(7.457 ± 0.398)kU/L, P ＜ 0.05 or ＜ 0.01], GSH-Px activity[(481.284 ± 43.785),(492.223 ± 16.474), (382.762 ± 25.167), (293.687 ± 24.881 )kU/L] was also lower than those of the control group [(585.078 ± 47.323)kU/L, P ＜ 0.05 or ＜ 0.01], MDA level[(0.609 ± 0.011 ), (0.646 ± 0.016), (0.852 ± 0.013),(1.188 ± 0.045)nmol/L] was higher than those of the control group[(0.512 ± 0.027)nmol/L, P ＜ 0.05 or ＜ 0.01];iNOS activity[(3.604 ± 0.115), (3.615 ± 0.075), (3.848 ± 0.103), (4.275 ± 0.079)kU/L] also was higher than those of the control group[(2.798 ± 0. 136)kU/L, all P ＜ 0.01], iNOS mRNA expression increased, eNOS activity [(5.539 ± 0.079), (5.503 ± 0.064), (5.226 ± 0.142), (4.809 ± 0. 107)kU/L] decreased compared to those of control group[(5.996 ± 0.155)kU/L, P ＜ 0.05 or ＜ 0.01], eNOS mRNA expression decreased; ICAM-1 levels [(0.852 ± 0. 102), (0.886 ± 0.061 ), (0.961 ± 0.158), (1.418 ± 0. 167)μg/L] increased compared to those of the control group[(0.687 ± 0.046)μg/L, P ＜ 0.05 or ＜ 0.01], VCAM-1 levels[(2.719 ± 0.197), (2.946 ± 0.167),(3.173 ± 0.225 ), (3.613 ± 0. 153 ) μg/L] was higher than those of the control group [(2.375 ± 0.067 ) μg/L, all P ＜0.01]. Conclusions High concentrations of fluoride reduce the activity of antioxidant enzymes, which leads to metabolic disorders of nitric oxide and abnormal cytokines expression, thereby inhibiting vascular endothelial cell growth, structural change and induced apoptosis. This is an important factor in high fluoride-induced vascular endothelial injury.

OBJECTIVETo investigate the correlation between the stage of hepatic fibrosis and ultrasonographic findings of the liver, spleen and gallbladder and establish a sensitive ultrasonographic semi-quantitative scoring system for screening compensated liver cirrhosis.

METHODSTotalling 248 patients with chronic hepatitis B and hepatitis C virus infection underwent liver biopsy and ultrasonic examination. The images of the liver surface, parenchymal echo, intrahepatic vessels, gallbladder, spleen and diameter of portal vein were analyzed.

RESULTSThe stages of hepatic fibrosis were not correlated to ultrasonographic findings of the liver surface or diameter of portal vein, but hepatic fibrosis of different stages showed significant differences in parenchymal echo, intrahepatic vessels, gallbladder and splenomegaly. In cases with normal liver parenchymal, intrahepatic vessels, gallbladder and spleen, the negative predictive value of the ultrasonographic semi-quantitative scoring system for diagnosing compensated liver cirrhosis amounted to 96.3%. The sensitivity of a score not lower than 5 was 90% for detecting compensated cirrhosis. With a score not lower than 7, the diagnostic accuracy and specificity was 85.9% and 95.2%, respectively, but the sensitivity was lowered to 37.5%.

CONCLUSIONThe ultrasonic images of the liver parenchyma, intrahepatic vessels, gallbladder and spleen in patients with compensated liver cirrhosis vary significantly in patients with hepatic fibrosis of different stages, and this ultrasonographic scoring system allows for a sensitive diagnosis of compensated cirrhosis.

Female ; Fibrosis ; Gallbladder ; diagnostic imaging ; Hepatitis B, Chronic ; complications ; Hepatitis C ; complications ; Humans ; Liver ; diagnostic imaging ; pathology ; virology ; Liver Cirrhosis ; complications ; diagnosis ; Male ; Reproducibility of Results ; Sensitivity and Specificity ; Spleen ; diagnostic imaging ; Splenomegaly ; diagnostic imaging ; Ultrasonography ; methods