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.

[Objective] To observe the therapeutic effect of Shengji Yuhong Ointment (SYO) in promoting healing of postoperative wound of tuberculous perianal abscess and anal fistula. [ Methods ] Forty cases of tuberculous perianal abscess and anal fistula after operation were divided into group A ( n = 20) and group B ( n = 20) according to the odd-or even-numbered days of admissions. Group A was treated with external application of SYO and group B with external application of vaseline ointment. After treatment, the healing of postoperative wound was observed in the two groups. [Results] In group A, the time for the ceasing of exudates from wound surface, for the formation of granulation and epithelial tissue, and for the healing of wound was shorter than that in group B ( P

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.

Treated 36 cases of infantile enuresis by acupuncturing Zuyunganqu (Foot Motor Sensory Area),Guanyuan (CV 4), Qihai (CV 6), Zhongwan (CV 12),Zusanli (ST 36), Yinlingquan (SP 9), Pishu (BL 20),Weishu (BL 21) and Shenshu (BL 23). After two courses,29 cases were cured, 5 cases were improvement and 2cases were no effect.

Objective To establish the quality standard for mongolian medicine Rhaponticum uniflorum (L.) DC. with modern analysis methods. Methods Chlorogenic acid and apigenin were identified by thin -layer chromatography (TLC). The chlorogenic acid content was determined by high performance liquid chromatograghy ( HPLC). And HPLC was performed on Kromat Universil C18 ( 4.6 mm × 250 mm, 5 μm) with methanol-water-acetic acid ( v/v/v, 15:85:0.85) as mobile phase, the detection wavelength was 326 nm, the flow rate was 1.0 mL/min, and the column temperature was 20℃. Results The samples and the reference substance shared the same color spots at the same sites. The linear range of chlorogenic acid was 0.044 66-0.446 6μg (r=0.999 8), and the average recovery rate was 99.84%. Conclusion The method is simple, accurate, and with good reproducibility, and can be used for the quality control of Rhaponticum uniflorum ( L. ) DC.

Cryptogenic stroke refers to ischemic stroke that can not be defined by routine examination at present. Studies have show n that patent foramen ovale may be a common cause of cryptogenic stroke, and its possible mechanism is the paradoxical embolism. With the development of imaging technology , more and more studies have show n that patent foramen ovale is closely associated w ith cryptogenic stroke. This article review s cryptogenic stroke in patients w ith patent foramen ovale.

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.

Guinea pig as a commonly used laboratory animal is widely used in various fields of biomedical research.The stability of genetic quality directly affects its development and application .Genetic testing is designed to confirm the genetic characteristics of each strain , to verify whether there are genetic mutations and other genetic contamination, to ensure that the test object meets the requirements of this strain .Along with the emerge of biochemical and molecular marker technology , a more convenient and reliable means is provided for research of genetic homozygosity , genetic type detection and genetic quality monitoring of guinea pigs .In this paper, the application and research progress of biochemical, cytological and molecular markers in studies of guinea pig diversity will be summarized , and provide some help for genetic testing guinea pig.