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.

BACKGROUND: Clusterin has many biological functions, especially the great protection effects on histiocytes in pathological conditions. It will be helpful to discuss the expression of clusterin in the spinal cord injury tissue to further identify the mechanism of secondary spinal cord injury and to provide possible treatments.OBJECTIVE: To observe the expression of clusterin in acute spinal cord injury tissue.DESIGN: Randomized controlled animal study.SETTING: Department of Orthopaedics, First Affiliated Hospital, China Medical University.MATERIALS: The experiment was performed in Experimental Animal Department, China Medical University from January 2003 to January 2006.A total of 65 adult healthy SD rats of 250-300 g provided by Experimental Animal Department, China Medical University were selected. METHODS: All the rat models by modified Allen assay for acute spinal cord injury were randomized into injury group, sham operation group and normal control group with 30, 30 and 5 rats in each group, respectively.The rat injury models in dorsal spinal cord were prepared by modified Allen assay: the target injury site was T10 segment; the sham operation group was given T10 total laminectomy. The normal control group received no operation. Rats in the injury group and sham operation group were killed at hour 12, days 1, 3, 7, 14 and 21 after spinal injury and operation with 5 rats at each time point, respectively. Frozen sections of the injury part were made at different time point after injury. Degeneration and necrosis of spinal cord injury tissues were observed with hematoxylin-eosin (HE) staining. Deposition and disposition of positive reactants of clusterin were observed with clusterin immunohistochemical staining. Average gray value of positive reactants of clusterin was detected with semi-quantitative image analysis (went with the intensity of clusterin immunoreaction in inverse ratio).MAIN OUTCOME MEASURES: ①Degeneration and necrosis as well as positive expression of clusterin of spinal cord injury tissues of rats in each group. ②Average gray value of clusterin positive reactants in spinal cord injury tissues of rats in each group.RESULTS: A total of 65 rats were involved in the result analysis, without dropout. ①In the injury group, positive expression of clusterin began to increase at day 1 after injury, reached peak at about day 7, declined gradually at day 14 and tended to be stable at day 21 after injury. This dynamic course of change was along with the prolongation of time. The expression of clusterin appeared in the sham operation group and normal control group at each time point, and the expression was stable. There was no dynamic change with the prolongation of time. ②In the injury group average gray value of positive reactants of clusterin was significantly lower at days 1, 3,7, 14 and 21 as compared with that in the sham operation group and normal control group (t=6.33-15.57, P ＜ 0.01 ). There was no significant difference in average gray value of positive reactants of clusterin at each time point in the sham operation group as compared with the normal control group (P ＞ 0.05).CONCLUSION: The expression of clusterin increases greatly in acute spinal cord injury tissues, and has dynamic change, which is the same with spinal cord injury.

Objective To understand and to assess the current situation of iodine deficiency disorders in Qaidam Basin,and to provide the information for measures and strategies of prevention and cure of this disease.Methods Within the boarder of Qaidam,five counties(cities)were selected for the investigation,which were Delingha,Golmud,Dulah,Wulan,and Tianjun.Two townships(towns)have been selected in each county(city),51 students aged 8 to 10 in one of the primary schools in each township(town)underwent thyroid palpation examination,their urine samples were collected to test urine iodine using arsenic-Ce catalysis spectrophotometric method.Thirty students of fifth-grade class in the selected schools underwent the questionnaire of prevention knowledge of iodine deficiency disease,while 10 housewives in each village near by the school were asked for the same questions,and the edible salt in their home was quantified of iodine by using direct titration method.Results Among the children aged 8 to 10,the average prevalence rate of thyroid enlargement Was 5.9%(30/510),higher than that stipulated by the national standard.The median of urinary iodine of urinary iodine level for children aged 8 to 10 was 83.4μg/L,the ration of urinary iodine＜20μg/L Was more than 10 percent;qualified iodized salt consumption rate of residents was 59.0%(118/200);the average scores and the checkout rate of fifth-grade students and the housewives who had underwent the questionnaire survey were 31.5,12.0%(36/300)and 35.9,18.5%(37/200)respectively.Conclusions The prevention and cure of iodine deficiency disorders in selected area do not work well,iodized salt is not popular,therefore health education needs to be strengthened.

Objective To investigate the differences of symptoms,physical signs and examinations in childhood and adult patients with Marfan syndrome.Methods Twenty-four children diagnosed as Marfan syndrome were investigated and evaluated synthetically in symptoms,physical signs and examinations.The positive rate of each item was analyzed to find the differences between childhood and adult patients.Results Among 24 patients,thorax and spinal deformity were found in 19 children(79%),leptosome type existed in 16 children(67%),dolichostenomelia was found in 15 children(63%).The dilation of the aortic sinus was detected in 16 children ((67%),)and the dilations of left ventricle and mitral regurgitation were found in 3 children(13%) with echocardiogram.Conclusions It is critical ascertain the abnormalities in cardiovascular system and give intervention,and it is expected to prolong the patients′ life by slowing down the changes in aortic walls.

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.

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.

This article reviews the moral judgment ability of patients with autism spectrum disorder (ASD),as well as the relationship with theory of mind (TOM).The study found that patients with ASD could undertake basic moral judgment,but could not provide the corresponding judgment reason,and were easy to make utilitarianism judgment.In addition,individual's moral judgment ability was positively correlated with TOM,while there were defects in TOM among the patients with ASD,which could lead to the skew results of moral judgment.