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.

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

Objective: To summarize the point selection pattern and treatment method in acupuncture-moxibustion for dyspepsia through data mining of the related clinical literatures in recent two decades, and to provide reference for clinical application. Methods: With data mining technology, clinical literatures related to acupuncture-moxibustion for dyspepsia published between January 1997 and July 2018 were collected from major Chinese databases, and the treatment methods, point and meridian frequencies were summarized and analyzed. Results: A total of 106 studies were included. For meridians, the top 5 meridians used were the Stomach Meridian, Conception Vessel, Spleen Meridian, Liver Meridian and Pericardium Meridian. For point selection, the leading 5 points were Zusanli (ST 36), Zhongwan (CV 12), Shangwan (CV 13), Xiawan (CV 10) and Sanyinjiao (SP 6). For major and adjunct points, the 5 most significant major points were Zusanli (ST 36), Zhongwan (CV 12), Neiguan (PC 6), Sanyinjiao (SP 6) and Shangwan (CV 13); the 5 most significant adjunct points were Neiting (ST 44), Qihai (CV 6), Fenglong (ST 40), Yinlingquan (SP 9) and Tianshu (ST 25). For treatment method, the top 3 methods used were moxibustion alone, acupuncture alone and acupuncture combined with medicinal herbs. Conclusion: The main meridian chosen in acupuncture-moxibustion for dyspepsia was the Stomach Meridian, and the points were mainly selected from the Stomach Meridian; treatment method was moxibustion alone, which may provide reference for clinical treatment of this disease.

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.

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 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 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.

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.