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 study the expression and its significance of bcl-2 associated death (bad) gene in human optic nerves from traumatic atrophic eyeballs. Methods The optic nerves from 8 normal human donor eyes and 31 traumatic atrophic eyes were studied by immunohistochemistry technique. Results Bad protein was positively expressed in the normal optic nerve myelin sheath and residual myelin portions of optic nerve tissues from traumatic atrophic eyes. The expression of bad protein in the residual portions of myelin sheath was stained significantly stronger than that in normal optic nerves (P0 05). Conclusion Bad might possess the function of promoting the optic nerve atrophy processes in traumatic atrophic eyes.

Infection can complicate different stages of acute pancreatitis.The probability of infection is related to the existence and extent ofpancreatic necrosis: up to 30% of patients with pancreatic necrosis develop infection.The timing of infection is variable and usually peaks in the second to fourth week after the onset of pancreatitis.IPN(infected pancreatic necrosis)is the most common type of secondary pancreatic infection.Infectious complications in severe acute pancreatitis are associated with considerable morbidity and mortality.Antimicrobial therapy is one of the fundamental elements of therapy.Diagnosis of secondary pancreatic infection is challenging and radiologic characteristic (air bubble sign) and CT guided fine-needle aspiration (FNA) are reliable diagnostic strategies.Antibioticuse should be rational in terms of a rational indication, a rational spectrum, and a rationalduration.Prophylactic antibiotics are not beneficial in improving prognosis.The only rational indication for antibiotics is documented infection.The empirical antibiotics should cover common microbials of hospital-acquired intra-abdominal infection and with good pancreatic penetration.Also, fungal infections are often present in thesepatients, and early diagnosis and antifungal coverage should be considered.Duration is in fact largely determined by the presence and efficacy of source control.When there is no or minimal residual infection left after a source control procedure, a duration of 7 to 10 days is probably sufficient.On the other hand, if the source of infection has not been removed completely, prolonged courses until the residual necrosis has been sterilized are necessary.

Objective To study the cutoff point of chronic nasal sinusitis with nasal polyps(CRSwNP) classified by the tissue eosinophils(Eos).Methods 140 CRSwNP patients were enrolled in this study.The data of their clinical feature were collectted before surgery.Mucosal specimens were assessed for the presence of tissue eosinophil cells.Follow up and record the data at 3 months and 6 months after surgery.Results When the tissue Eos grouped by 50/HPF, the VAS, nasal sinus CT ,endoscopic preoperative score,and the endoscopic postoperative score at 3 months and 6 months showed significant difference(P<0.05).The tissue Eos between skin prick test(SPT) positive group and SPT negative group showed no significant difference.The tissues Eos was sighificantly correlated with the peripheral Eos(ρ=0.459,0.473,P<0.01).Conclusion The most meaningful cutoff point may be 50 Eos/HPF nearby to distinguish ECRS and non-ECRS(taken nasal polyps,by random HPF).Allergic factors may have no effect or little effect on the increase in Eos of nasal polyps in patients with CRSwNP;peripheral blood Eos may be an indicator of the level of Eos in tissues.

[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 make sure who is liable for Adverse Drug Reactions(ADRs)and what kind of legal obligation should they bear and how to bear.METHODS:The concerned laws and regulations were studied by literature research method and comparative study method.RESULTS:The legal obligations involved in ADRs were tort liability and administrative liability.The obligation attribution principles were different.The fault principle should be applied to the unpredicted“develop_ mental danger”of drugs.CONCLUSION:The compensatory and relieving acts related to ADRs damages should be formulated in China.

Objective To investigate the effects of inhibitor of growth 4(ING4)on the proliferation,migration and the expression of angiogenesis related factors such as VEGF,MMP-2,MMP-9 in endothelial cells. Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro;ING4 plasmid and siRNA were constructed and transfected to HUVECs;the proliferation of HUVECs was evaluated by MTT assay;the ability of migration was evaluated by Transwell assay;real-time PCR and Western blotting were used to determine the expression of mRNA and pro-tein of angiogenesis related factors such as VEGF,MMP-2,and MMP-9. Results MTT and Transwell assay showed that ING4 has the ability to inhibit the proliferation and migration of HUVECs,and the results of real-time PCR and Western blotting proved that ING4 can inhibit the expres-sion of angiogenesis related factors such as VEGF,MMP-2,and MMP-9. Conclusion ING4 can inhibit the proliferation and migration of HU-VECs,down-regulate the expression of angiogenesis related factors such as VEGF,MMP-2,MMP-9,and inhibit angiogenesis.

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.