Objective To explore the risk factors of primary nephritic syndrome complicated with thrombosis in children. Methods Clinical data of 238 children with primary nephritic syndrome were retrospectively analyzed. The children were divided into thrombosis group and non-thrombosis group according to whether complicated with thromboembolism. Univariate and logistic regression analysis were performed.Results Among 238 children, there were 32 cases of primary nephritic syndrome complicated with thrombosis and the rate was 13.44%. Univariate analysis showed that infections, the use of diuretic, degree of edema, white blood cell count, IgG, C3, total protein, albumin, urea nitrogen, plasma ifbrinogen, D-dimer, antithrombinⅢ, and 24-hour proteinuria were signiifcantly different between two groups (allP< 0.05). Multivariate analysis showed that D-dimer and 24-hour proteinuria were the independent risk factors for children with primary nephrotic syndrome complicated with thrombosis.Conclusion The elvated level of D-dimer and 24-hour proteinuria were the risk factors of children with primary nephrotic syndrome complicated with thrombosis.

Objective To explore the effects of BCG and Poly I:C co-vaccination on the development of spleen T cell subsets of neonatal BALB/c mice. Methods Neonatal BALB/c mice were inoculated with BCG and/or Poly I:C intraperitoneally within 2-3 d after birth. Four weeks later, spleen cells of mice were isolated and the percentage of CD3+ CD8+ IFN-γ+,CD3+ CD8-IFN-γ+,CD3+ CD8+ IL-4+,CD3+ CD8- IL-4+,CD4+ Foxp3+ T cells,which represent Tc1,TH1,Tc2,TH2,Treg cells,respectively,were tested by flow cytometry at single cell level,and the ratios of TH 1/TH 2 and Tc1/Tc2 were calculated. Results The percentages of TH1 and Tc1 cells of BCG-vaccinated mice,Poly I:C-vaccinated mice and BCG plus Poly I:C-vaccinated mice were significantly higher than that of control mice(P＜0.05 or P＜0.01),and there was no difference among the three vaccinated group. The ratios of TH1/TH2 and total IFN-γ/IL-4 of the three vaccinated groups were higher than that of control group,but not the ratio of Tc1/Tc2. The TH1/TH2 ratio of BCG plus Poly I:C-vaccinated group was higher than that of BCG-vaccinated group(P＜0.05).The percentages of Trge cells showed no difference among the four groups(P＞0.05). Conclusion BCG and Poly I:C co-vaccination can significantly increase the number of Tc1 and TH 1 cells and TH 1/TH2 ratio in spleen cells. BCG and Poly I:C vaccination may have a synergistic effect on TH 1/TH2 ratio of spleen cells in neonataI mice. The percentage of CD4+ Foxp3+ T cells among four groups showed no significant difference.

Objective To construct a eukaryotic expression vector in Pichia pastoris containing human tissue factor( hTF) gene,in order to achieve high level secretory expression in extracellular.Methods Expression plasmid, pGAPZaA-hTF, was constructed by inserting the synthesized sequence encoding human extracellular tissue factor into yeast expression vector pGAPZaA and transformed into Pichia pastoris SMD1168H with electroporation.Having been selected by Zeocin, transformants containing hTF cDNA were expressed in YPD, extracellular proteins were detected by SDS-PAGE and confirmed by Western bolt.Results Successfully constructed the recombinant pGAPZaA-hTF expression system in Pichia pastoris.SDS-PAGE showed that the molecular weight of the expression product was about 37 -40 kDa.Western-blot indicated that it was human extracellular tissue factor.The crude yield of total protein in medium was up to 1 g/L, more than 80% of which was hTF.Conclusion Truncated rTF gene is expressed in Pichia pastoris and the active products are secreted into the medium which have the same activation as the native TF.

Pullulan is a linear glucosic polysaccharide produced by the polymorphic fungus Aureobasidium Pullulans, which has long been applied for various applications in medical and food industry due to its security, stability and low adhesive ability.At present, the two problems in restricting pullulan industrial production are the low polysaccharide production and melanin secreted which is hard to erase completely, giving the following process some problem.As a starting point, this review article collects and analyzes the progress on the breeding of pullulan high-yield strain without melanin in recent years, in order to find more efficient strains breeding methods, laying a foundation for further breeding of pullulan high-yield strain without melanin.

ObjectiveTo explore the changes and significances of cardiac troponin Ⅰ (cTnⅠ),myoglobin (Mb) and creatine kinase MB (CK-MB) in old patients with acute myocardial infarction(AMI).MethodThe levels of cTnⅠ, Mb and CK-MB in 89 cases with AMI patients (AMI group) and 100 healthy controls (control group) were detected and compared. ResultsThe levels of cTnⅠ, Mb and CK-MB in AMI group were ( 8.15 + 3.26) μ g/L, (478.45 ± 96.87 ) μ g/L and ( 128.17 ± 53.26 ) U/L, while were (0.03 ±0.02) μ g/L, (21.61 + 9.38 ) μ g/L and (9.53 ± 2.94) U/L in control group, the levels of cTnⅠ, Mb and CK-MB in AMI group were significantly higher than those in control group(P ＜ 0.05 ). The sensitivity rate of cTnⅠ,Mb and CK-MB for detecting AMI was 95.5％ (85/89), 97.8％ (87/89) and 87.6％ (78/89), and the specificity rate was 98.0％(98/100), 82.0％(82/100) and 94.0％(94/100). ConclusionThe levels of cTnⅠ, Mb and CK-MB are significantly increased in AMI patients, which cTnⅠ for detecting AMI has high sensitivity and specificity.

Nisin, produced by several strains in the growth process of Lactococcus lactis, is a natural antimicrobial polypeptide.Now, Nisin has served as an effective and safe food additive extensively used in food industry in many countries and regions because of its excellent antimicrobial activity.However, the current production of Nisin is largely fermented by lactobacillus and its industrialized production still can not meet enormous market needs, therefore establishing reasonably high-yield Nisin strains is of great significance.This review mainly summarizes the development pathway of molecule based on the functional expression of Nisin biosynthetic genes and regulation of gene expression, and also the study status on high Nisin-producing strains which provides practical foundation for further study on expected strains as well as some useful guidance for large-scale industrialized production of Nisin.

BACKGROUND:Tissue engineering provides new ideas and approaches for repair of cartilage defects.
OBJECTIVE:To develop a complete set of solutions for construction of tissue engineered cartilagein vitro, with chondrocytes as seed cels and cross-linked sodium hyaluronate as scaffold materials.
METHODS: New Zealand rabbit articular chondrocytes were isolated, counted, and then cultured and passaged to prepare cellsuspension. Toluidine blue staining, RT-PCR and immunocytochemistry were exerted to evaluate the cultured cels. Chondrocytes were seeded and co-cultured with cross-linked sodium hyaluronate scaffold for 21 days. Then, RNA was isolated for RT-PCR, and frozen sections were prepared for morphological observation and immunohistochemistry study.
RESULTS AND CONCLUSION:The chondrocytes could adhere to the cross-linked sodium hyaluronate scaffold and aggregate, growing between fibers or adhering to the scaffold in a monolayer manner. The transcripts of cartilage specific aggrecan gene and colagen type II alpha 1 gene and cartilage specific protein colagen type II were expressed in cel-scaffold complexes to maintain the phenotype of chondrocytes. Cel-scaffold complexes co-cultured in vitro can form cartilage extracelular matrix, by which tissue engineered cartilage is expected to be obtained.

BACKGROUND:Salvia miltiorrhiza bone-setting capsule is a traditional Chinese medicine for the treatment of fractures due to activating blood circulation to dissipate blood stasis, reducing sweling and pain. OBJECTIVE: To observe the effects of Salvia miltiorrhiza bone-setting capsule on the fracture healing in a rat model of closed femoral fractures. METHODS: Rats were randomly divided into salvia miltiorrhiza bone-setting capsule group, physiological saline group and normal group. In the salvia miltiorrhiza bone-setting capsule group and physiological saline group, rat models of closed femoral fractures were prepared, and then given physiological saline and salvia miltiorrhiza bone-setting capsule 2 pils by intragastric administration. In the normal group, rats were housed normaly. At 7, 14 and 28 days after fractures, hematoxylin-eosin staining conditions, serum osteocalcin, the expression of colagen type I, and the expression of protein and mRNA calus transforming growth factor-beta 1 were observed in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining demonstrated that at 7 days after fractures, no significant difference was found in pathological changes of femoral fracture in salvia miltiorrhiza bone-setting capsule group and physiological saline group. At 14 and 28 days after fractures, pathological repair was more obvious in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group. (2) At 3 and 7 days after fractures, serum osteocalcin and the expression of type I colagen were significantly increased in the salvia miltiorrhiza bone-setting capsule group and physiological saline group (P < 0.05), and the expression trend was consistent in both groups. The expression was always higher in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group, and significant differences were found at 14 and 28 days after fractures (P < 0.01). (3) Transforming growth factor beta 1 expression reached a peak at 3 days after fractures, gradualy reduced, increased at 14 days (the second peak), and diminished at 28 days in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. The expression trend of transforming growth factor beta 1 was consistent in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. At 7, 14 and 28 days, the transforming growth factor beta 1 expression was higher in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group. (4) Results showed that salvia miltiorrhiza bone-setting capsule could promote fracture healing, and its mechanism was probably associated with serum osteocalcin, the expression of colagen type I and transforming growth factor-β1.

Objective To construct a eukaryotic expression vector in Pichia pastoris containing human fibrinogen gene, in order to achieve high level secretory expression in extracellular.Methods Expression plasmid,pGAPZαA-FGB-FGG-FGA-AOX1,was constructed by inserting the synthesized sequence encoding human fibrinogen(FGA, FGB,FGG) and then introduced into Pichia pastoris SMD1168H by electroporation.Transformants were availably screened by Zeocin resistance,the expression of recombinant protein was identified by SDS-PAGE and Western blot analysis, the protein yield was tested by ELISA assay.After ultrafiltration and purification, the biological activity of protein was detected.Results The crude yield of human fibrinogen in Pichia pastoris supernatant reached 15 mg/L in flask and the biological aggregation activity was determined.Conclusion The human fibrinogen gene was obtained and successfully expressed in Pichia pastoris and the active products were secreted into the medium.

Objective To evaluate the effects of fluticasone propionate (FP)on Foxp3 expression in CD4+T cells, to explore the possible mechanisms of childhood asthma. Methods Thirty asthmatic children, 15 with inhaled FP and 15 without inhaled FP, and 16 healthy children were recruited. Peripheral blood mononuclear cells (PBMC)labeled for CD4 and intracellular Foxp3 were analyzed using flow eytometry. The levels of IL-2 and IL-6 in serum and supernatant before and after stimulation by Phytohemagglutination (PHA)were measured by ELISA. The expression of phosphorylated signal transducer and activator of transcription 5 (STAT5)in PBMC was detected by Western-blot.Results Compared with healthy control, the percentage of CD4+ Foxp3+ cells in PBMC in asthmatic children without inhaled FP was significantly decreased. After inhaled FP and in remission stage, the percentage of CD4+ Foxp3+ cells in asthmatic children was up regulated with a decreased serum IL-6 level and an increased phosphorylated STATS expression. Conclusions Decreased Foxp3 protein expression in peripheral CD4+ T regulatory cells (Treg)is characterized in childhood asthma. Inhaled glucoeorticoid therapy of childhood asthma might be attributed to its ability of increasing Foxp3 expression by upregulation of phosphorylated STAT5 to balance the T cell response.