Aim To investigate whether liver X recep- tors attenuate high glucose-induced apoptosis in H9C2 cells through inhibiting nuclear factor-NF-κB.Methods The lentiviral vector of LXRs was constructed and H9C2 cells cultured in high glucose were infected.The H9C2 cells were divided into 6 groups:control group (5.5 mmol·L -1 glucose),mannitol group(5.5 mmol ·L -1 glucose +27.5 mmol·L -1 mannitol),high glu-cose group(33 mmol·L -1 glucose),green fluorescent protein(GFP)group LXRαgroup,and LXRβgroup. The inhibition rate of H9C2 cells,the mRNA of Bax, Bcl-2,the protein content of NF-κB,Bax,Bcl-2, cleaved caspase-3,and the cell apoptosis were com-pared among these groups.Results LXRs overexpres-sion significantly attenuated high glucose-induced in-crease in Bax NF-κB,cleaved caspase-3 and cell ap-optosis(P <0.05),and increased high glucose-induced decrease in Bcl-2.Conclusion Liver X receptors at-tenuate high glucose-induced apoptosis in H9C2 cells through NF-κB signaling pathway.

AIM: To explore the histocompatibility of pinealocyte microencapsules in vivo. METHODS: The pineal glands of neonatal rats were removed under operating microscope and pinealocytes were isolated through collagenase and trypsin digestion. Pinealocytes were cultured for one week in vitro and collected immediately after digesting was encapsulated in APA microencapsules. The cells and empty capsules were transplanted into abdominal cavity or intermuscular space respectively and retrieved at the 15th or 30th day after operation. Morphological observation, HE staining, cell counting, and HPLC technique were used to analyze the shape, proliferation and function, the degree of inflammation fibrosis of retrieved microencapsules. RESULTS: The retrieve rate of cell capsule from abdominal cavity was about 85 % . The retrieved capsules had integrated profile mostly although some were damaged. The amount of macrophages attached to capsule wall and the thickness of wall increased gradually following the period of transplantation. However, the retrieve rate, wall thickness had no difference between retrieved cell and empty capsules at the same time. Secretion ability of pinealocytes in capsule retrieved at 15th day after operation decreased rapidly and those retrieved at 30th day after operation lossed secretion function. CONCLUSION: APA microencapsules had histocompatibility relatively in vivo and protected pinealocytes in capsule from immunologic rejection of the host. The survival time was about 20 days. During this period cells in capsule maintained activity and MT secretion ability.

Objective To investigate the relationship between the coagulation system status and the pulmonary hemorrhage in children with severe hand,foot and mouth disease(HFMD)and approach the clinical significance of early detection of coagulation function. Methods By prospective case design method,89 cases with HFMD admitted to Department of Critical Care Medicine of Hebei Provincial Children Hospital from July 2010 to July 2012 were enrolled. The children were divided into severe group(46 cases)and critical group(43 cases)according to the severity of disease,and the children in critical group were subdivided into survivor group(26 cases)and non-survivor group (17 cases). Forty-four healthy children with the same age and in the same period were served as healthy control group. The blood of children was collected immediately after admission for determination of blood routine, prothrombin time(PT),thrombin time(TT),activated partial thrombin time(APTT),fibrinogen(Fg),and D-dimer (DD). Results There were no significant differences in PT,TT,APTT and Fg among severe group,critical group and health control group(all P>0.05). The blood platelets count(PLT)in severe group and critical group was significantly lower than that in health control group(×109/L:245±130,237±156 vs. 389±120),while the DD was significantly higher than that in healthy control group(mg/L:0.34±0.67,0.41±0.08 vs. 0.24±0.13),and the DD in critical group was obviously higher than that in severe group(all P<0.05). The mortality rate in critical group was 39.5%,and there were no significant differences in PT,APTT,Fg,TT and PLT between survivor group and non-survivor group(all P>0.05),but the DD in non-survivor group was significantly lower than that in survivor group(mg/L:0.60±0.09 vs. 0.12±0.09,P<0.05). Conclusions In children with severe or critical HFMD, the coagulation factor and blood platelet were in a state of mobilization,mild consumption state with the existence of fibrinolytic inhibition,but without systemic bleeding tendency,therefore it is in a compensatory stage of disseminated intravascular coagulation(DIC),not the mechanism of pulmonary hemorrhage. The monitor of DD has its clinical significance in evaluations of the disease situation and its prognosis.

Halohydrin dehalogenase is of great significance for biodegradation of the chlorinated pollutants, and also serves as an important biocatalyst in the synthesis of chiral pharmaceutical intermediates. A putative halohydrin dehalogenase (HheTM) gene from Tistrella mobilis KA081020-065 was cloned and over-expressed in Escherichia coli BL21 (DE3). The recombinant enzyme was purified by Ni-NTA column and characterized. Gel filtration and SDS-PAGE analysis showed that the native form of HheTM was a tetramer. It exhibited the highest activity at 50 degrees C. The nature and pH of the buffer had a great effect on its activity. The enzyme maintained high stability under the alkaline conditions and below 30 degrees C. HheTM catalyzed the transformation of ethyl(S)-4-chloro-3-hydroxybutyrate in the presence of cyanide, to give ethyl (R)-4-cyano-3-hydroxybutyrate, a key intermediate for the synthesis of atorvastatin.
3-Hydroxybutyric Acid ; chemistry ; Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Escherichia coli ; Hydrolases ; genetics ; metabolism ; Hydroxybutyrates ; chemistry ; Recombinant Proteins ; genetics ; metabolism ; Rhodospirillaceae ; enzymology ; genetics

ABSTRACT OBJECTIVE：To compare the effectiveness and economics of peramivir and oseltamivir in the treatment of influenza complicated with febrile seizures in children. METHODS：In retrospective study，152 children with influenza complicated with febrile seizures were collected from our hospital during Dec. 2018 to Mar. 2019.They were divided into peramivir group（81 cases） and oseltamivir group （71 cases）. Fever remission time， medication duration， hospital duration， clinical efficacy （determined by convulsion，cough，nasal obstruction，runny nose，sore throat，etc.），the incidence of rash，the number of children with combined antibiotic and TCM were compared between 2 groups. Cost-effectiveness analysis was used to evaluate cost-effectiveness ratio（CER）and incremental cost-effectiveness ratio（ICER）of medication regimen in 2 groups. The sensitivity analysis of the effect and total cost were carried out with Logistic regression and multiple linear regression respectively through CER and ICER calculated by reducing 15％ drug price. RESULTS：There was no statistical significance in fever remission time， medication duration，hospitalization duration，the incidence of rash and proportion of children with combined antibiotics between 2 groups（P＞0.05）. There was statistical significance in proportion of children with combined TCM and clinical effect，and the proportion of children with combined TCM in peramivir group was significantly lower than oseltamivir group（P＜0.001）. Clinical effect of peramivir group was significantly better than that of oseltamivir group（P＝0.021）. Total cost of peramivir group and oseltamivir group were 5 442.84 yuan/person and 5 571.71 yuan/person（P＝0.795）；CER of them were 54.47 and 56.51；ICER of peramivir group was － 89.38. The results of sensitivity analysis were consistent with those of basic analysis.CONCLUSIONS： Compared with oseltamivir，peramivir is more effective and less costly for children with influenza and febrile seizures.

Inhibitor of nuclear factor kappa-B kinase subunit beta (IKKβ) is one of important kinases in inflammation to phosphorylate inhibitor of nuclear factor kappa-B (IκBα) and then activate nuclear factor kappa-B (NF-κB). Inhibition of IKKβ has been a therapeutic strategy for inflammatory and autoimmune diseases. Here we report that IKKβ is constitutively activated in healthy donors and healthy Ikkβ ^C46A (cysteine 46 mutated to alanine) knock-in mice although they possess intensive IKKβ-IκBα-NF-κB signaling activation. These indicate that IKKβ activation probably plays homeostatic role instead of causing inflammation. Compared to Ikkβ ^WT littermates, lipopolysaccharides (LPS) could induce high mortality rate in Ikkβ ^C46A mice which is correlated to breaking the homeostasis by intensively activating p-IκBα-NF-κB signaling and inhibiting phosphorylation of 5' adenosine monophosphate-activated protein kinase (p-AMPK) expression. We then demonstrated that IKKβ kinase domain (KD) phosphorylates AMPKα1 via interacting with residues Thr183, Ser184, and Thr388, while IKKβ helix-loop-helix motifs is essential to phosphorylate IκBα according to the previous reports. Kinase assay further demonstrated that IKKβ simultaneously catalyzes phosphorylation of AMPK and IκBα to mediate homeostasis. Accordingly, activation of AMPK rather than inhibition of IKKβ could substantially rescue LPS-induced mortality in Ikkβ ^C46A mice by rebuilding the homeostasis. We conclude that IKKβ activates AMPK to restrict inflammation and IKKβ mediates homeostatic function in inflammation via competitively phosphorylating AMPK and IκBα.