2,5-DKG reductase I was purified from cell-free extracts of a recombinant,ER97 by a procedure involving ammonium sulfate precipitation and successive column chromatography on DEAE-Sepharose CL-6B and Phenyl Sepharose CL-4B with 5 fold purification,27 % recovery and 3418 U/mg specific activity.The molecular weight of the enzyme estimated by SDS-PAGE was 34kD.The isoelectric point was estimated to be 6.0 by PAG-IEF.The optimum pH was 7.0 and the optimum temperature was about 40℃.The enzyme can catalyze the stereospecific NADPH-dependent reduction of 25-DKG to 2-KLG.The michaelis-menten constant(Km) for 2,5-DKG and NADPH were 0.29 mmol/L and 14.7 mmol/L respectively.The enzyme is specific for NADPH and 2,5-DKG,1 mmol/L Cu~(2+) or Zn~(2+) could highly inhibited the enzyme activity.EDTA and ?-Mercaptoethanol have no effect on the enzyme activity.

Because of the neonatal immune system is immature,particularly in preterm and low birth weight infants,the neonate is prone to some infections.Neonatal sepsis usually has the slight performance and atypical clinical symptoms,which raise the great interest of searching for some better biomarkers.With the development of testing technology,the research of serum inflammation marker has made remarkable progress at home and abroad in recent years,however,the research of umbilical blood inflammation marker is still in the early stage of exploration.Umbilical blood detection have the advantages of early,noninvasive,safe and convenient compare to the traditional serum detection.This paper mainly discusses the application of umbilical blood detection to early onset neonatal sepsis.

Objective: To analyze the constituents and metabolites of Radix Sa poshnikoviae (RS) in rat plasma and urine by rapid-resolution liquid chromatography-time of flight mass spectrometry (RRLC-TOF/MS), so as to explore the active ingredients and metabolites of RS in vivo. Methods: The separation was performed on a Angilent Zorbax Extend-C14 (5 μm, 250 mm x 4.6 mm id) column, with a methanol-water mobile phase system used for gradient elution. Time-of-flight mass spectrometer (TOF/MS) was applied for qualitative analysis under positive ion mode. Based on the accurate molecular weight of TOF/MS detection and the compound list of RS established previously, the constituents and metabolites of RS in different matrix in vivo were identified. Results: Six constituents of RS were identified in the plasma, sucrose, prim-O-glucosylcimifugin, cimifugin, nodakenetin, 5-O-methylvisamminol, and 3′-O-i-butyrylhammaudol. Eight constituents were identified in the urine, prim-O-glucosylcimifugin, divaricatacid, cimifugin, 4′-O-glucosyl-5-O- methylvisamminol, (3S)-2,2-dimethyl-3, 5-dihydroxy-8-hydroxymethyl-3, 4-dihydro-2H, 6H-benzo-[1, 2-b: 5, 4-b′] dipyran-6-one, 5-O-methylvisamminol, see-O-β-D-glucosylhammaudol, and wogonin. Two metabolites were identified in the urine, glucuronide of cimifujin and an isomer of it. Conclusion: The present method is reliable and effective for identifying compounds of RS in vivo, and it can provide a reference and evidence for the further pharmacodynamics experiments.

Humans ; Tuberculosis, Hepatic

Diagnostic Errors ; Dyspnea ; diagnosis ; Female ; Humans ; Infant ; Male

Objective To investigate the effect of lipoteichoic acid of bifidobacterium (BLTA) on the expressions of Fas/FasL in bearing melanoma B16 cancer mice. Methods Forty C57BL/6 mice were randomly divided into 4 groups (n=10),normal saline group (NS group),low-dose BLTA group (50 mg/L),middle-dose BLTA group (100 mg/L),and high-dose BLTA group (150 mg/L). Melanoma B16 cells (0.2 ml 5?109/L) were subcutaneously injected into the mice of the later 3 groups,and then,0.2 ml BLTA at corresponding doses was injected subcutaneously at the sites around the mass once a day for 10 d when the tumor (at least 2 mm?2 mm) was palpable. The NS group was given normal saline at same volume as in BLTA group. The changes of the killing activity of CTL was examined by MTT assay. The expression of the tumor-infiltrating CD4+ and CD8+ lymphocytes in tumor tissue were detected by immunohistochemistry. RT-PCR was used to detect the mRNA expression variance of the Fas and FasL in tumor and spleen tissue. The protein expression of Fas,FasL in tumor tissue and lymphocytes of spleen were detected by immunohistochemistry and Western blotting. Results Compared with the control group,BLTA significantly improved the CTL killing activity (P0.05). Conclusion BLTA enhances the killing activity of CTL by regulating Fas/FasL system,and thus,promotes apoptosis of tumor cells to suppress tumor immune escape in tumor-bearing mice,thereby it exerts anti-tumor effect further.

[Summary] Glucagon-like peptide-1 ( GLP-1 ) as a new treatment of type 2 diabetes, not only has hypoglycemic effect, but also plays an important role in the regulation of lipid metabolism. GLP-1 plays a unique role in regulating lipid metabolism via lipid absorption and transport, fat formation and decomposition, hepatic lipid metabolism, and cholesterol transport.

Objective To explore the immunogenicity of a novel bionic scaffold of nucleus pulposus tissue engineering. Methods The biocompatibility of a subcutaneously implanted scaffold of nucleus pulposus tissue engineering was studied in SD rats by analyzing tissue reactions up to 3 months using histological and ultrastructural methods. The expression of IFN-?, IL-2, IL-4 and IL-10 mRNA was measured by RT-PCR, and the levels of serum antibodies to porcine type Ⅱ collagen were measured by ELISA. Results There was less inflammatory reaction in rats induced by subcutaneously implanted scaffold. By degrees, a granulation tissue had developed within the implant, which had disappeared by 3 months. The expression of IFN-?, IL-2 mRNA by RT-PCR was of no changes. But the expression of IL-4 and IL-10 mRNA increased, which meant the implant induced Th cell into Th2 cell and the induction inhibited the inflammatory reaction. No antibodies to porcine type Ⅱ collagen were found in the sera of implanted rats. Conclusion There was less immunogenicity in rats induced by implanted scaffold of nucleus pulposus tissue engineering.

Objective To evaluate the performance of a latex enhanced immunoturbidimetric?2-microglobulin kit in clinical use. Methods The lower limit of detection,precision,accuracy and linearity range was determined on the automatic chemistry analyzer Hitachi 7170S. Results The lower limit of detection was 0. 09mg/L. The within-run precision was less than 4% while the between-run precision was less than 8%. The accuracy was 99.29 % and the linearity range was 0.2～18mg/L. Conclusion The?2-microgiobulin kit shows good performance in clinical use.

OBJECTIVE:To provide reference for the improvement on the management of defective medicinal products in China.METHODS:The current UK Defective Medicinal Products Guide and drug recall cases were analyzed and its implication on China was discussed.RESULTS & CONCLUSIONS:The mature defective medicinal products management experiences in UK serve as a mirror for drug safety work in China.At present,the drug safety work can be intensified through clarifying the definition of defective drugs,establishing effective information exchange channel for the defective drugs,establishing evaluation system about records of drug complaints,formulating scientific operational strategies for the recall of defective medicinal products and establishing matched information distribution channel.