BACKGROUND: The mitofusin 2 (Mfn2) may affects vascular smooth muscle cell Ras protein and suppress cellular proliferation through inhibition of extracellular signal-regulated protein kinase signaling pathway, which plays an important role in pathogenesis of vascular disorders such as hypertension, atherosclerosis and post-angioplasty restenosis. Mfn-2 gene amino acid sequence of the first 442 serine serves as protein kinase A (PKA) phosphorylation site, which is closely related to its phosphorylation status and may be involved in its functional regulation.OBJECTIVE: To investigate the effect of Mfn2 gene with PKA phosphorylation site deletion[Mfn2-PKA (△)] on inhibiting the proliferation of vascular smooth muscle cells and related signaling pathway.METHODS: Vascular smooth muscle cells of rats infected by recombinational adenovirus carrying green fluorescent protein,Mfn2 gane and Mfn2-PKA (△), were subcultured for 3-10 passages and randomly divided into 4 groups: ① Control group without intervention. ② Control group infected with adenovirus carrying green fluorescent protein. ③ Experiment group infected with adenovirus carrying Mfn-2 gene.④ Experiment group infected with adenovirus carrying Mfn2-PKA (△). Laser confocal microscopy was used to observe the locations of Mfn2 gene with and without PKA in cells. The expressions of extracallular signal-regulated protein kinase, Mfn2 gone and Mfn2-PKA (△) were determined by Western blot analysis. The growth curve of the vascular smooth muscle cells was explored by MTT.RESULTS AND CONCLUSION: The Mfn-2 and Mfn2-PKA (△) both expressed protein-specific bands in vascular smooth muscle cells. Two kinds of gone expression products were mainly located at the out membrane of mitochondria. Compared with the control group and adenovirus carrying green fluorescent protein group, the absorbance values at 3, 4, 5, 6 days were significantly reduced in adenovirus carrying Mfn2 group (P ＜ 0.01), and no obvious changes were observed in adenovirus carrying Mfn2-PKA (△) group. Compared with the control group and adenovirus carrying green fluorescent protein group, the extracellular signal-regulated protein kinase expression was significantly reduced in adenovirus carrying Mfn2 group (P ＜ 0.01), and no obvious changes were observed in adenovirus carrying Mfn2-PKA (△) group. Mfn2-PKA (△) located at the out membrane of mitochondria, has no effect on suppressing the proliferation of vascular smooth muscle cells, and no inhibition effect on extracellular signal-regulated protein kinase signaling pathway.

Objective To explore the effectiveness and safety of high frequency oscillatory ventilation(HFOV) in children with measles complicated with severe pneumonia and the acute respiratory distress syndrome(ARDS).Methods A total of 63 children with measles complicated with severe pneumonia and the ARDS were divided into conventional mechanical ventilation(CMV) group and HFOV group.The PaO2/ FiO2,oxygenation index (OI),HR and mean arterial pressure (MAP) before treatment and 12 h,24 h,48 h after treatment were detected.The rate of air leak and the motality in two groups were compared.The efficacy and safety of HFOV treatment were evaluated in children with measles complicated with pneumonia and severe ARDS.Results In HFOV group,the PaO2/FiO2 ratio was elevated and OI was decreased significantly after 12 h and maintained for at least 48 h.Compared with CMV group,OI of HFOV group improved more significantly,and the difference was statistically significant.The ventilation time in HFOV group was shorten than that in CMV group[(7.97 ±3.06) d vs.(11.03 ±3.60) d],but there was no statistical difference between two groups (P ＞ 0.05).The heart rate after treatment 48 h was gradually returned to normal,and there was no statistical difference between the two groups.There were no significant changes in the MAP of two groups after treatment.There were no significant differences in the incidence of air leak between the CMV group and the HFOV group(24.2％ vs.16.7％).The mortality rate of CMV group and HFOV group was respectively 45.5 ％ and 33.3 ％,and there was no statistically significant difference.Conclusion HFOV was effective in oxygenation and seems to be safe for pediatric patients with measles complicated with severe pneumonia and the ARDS.Also it didn't influence the occurrence of complications.It has no adverse influence on hemodynamic parameters.Early intervention of HFOV is safe and effective for the children with measles complicated with severe pneumonia and ARDS.

Objective:To develop an indirect enzyme-linked immunosorbent assay (ELISA) for semi-quantification of major allergen parvalbumin in fish.Methods:The soluble proteins were prepared from both white and dark muscles of seven species of freshwater fish and five species of marine fish.Tricine-SDS-PAGE and Western blot were performed to examine the protein patterns of fish muscle extracts.Natural parvalbumin being used to make calibration curve was purified from silver carp (Hypophthalmichthy molitrix) by ammonium sulphate fractionation,followed by ion exchange and gel filtration chromatography.The molecular mass of purified protein was estimated by Tricine-SDS-PAGE and identified by Western blot with anti-frog parvalbumin monoclonal antibody PARV-19.ELISA using PARV-19 was carried out to evaluate parvalbumin contents in white and dark muscles.Results:Tricine-SDS-PAGE revealed species-specific differences in proteins of heated extracts.Western blot confirmed that the major bands were showed in Tricine-SDS-PAGE with the molecular masses of 10-14 kD corresponded to parvalbumins recognized by PARV-19 and various numbers of isoforms of parvalbumin existed in different species of fish.There might be some differences in the parvalbumin contents and the epitope region was recognized by PARV-19 based on the differences in relative intensities of protein immunodetection.The ELISA showed that the contents of parvalbumin were 4 to 33 folds higher in the white muscle than in the dark muscle and varied greatly in different species of fish.Conclusion:These results validate that the dark muscle might be less allergenic than the white muscle due to the lower content of parvalbumins,and it is suggested that the commercial anti-parvalbumin antibody PARV-19 can be used to detect parvalbumins from the commercially important species of fish tested in this study and the method we develope succeeds to detect the major allergen in various species of fish.

Objective To study the effect of interleukin (IL)-10 knockout (IL-10-/-) on renal repair after renal ischemia-reperfusion injury in mice.Methods Eighteen IL-10-/-mice (KO) aged 8-10 weeks and 18 C57BL/6 wild type mice (WT) aged 8-10 weeks were divided into control group (Sham) and renal ischemia-reperfusion injury (IRI) group.The renal tissue morphology change was observed by Hematoxylin and eosin (HE) staining and Masson staining.The expressions of IL-18, Ki67 and TGF-β1 were detected by immunohistochemistry.The expression of TGF-beta1 and IL-18 were detected by Western blotting.Results Compared with that in WT-IRI group, in KO-IRI group renal pathological damage was more severe, renal interstitial fibrosis was visible, Ki67 expression of renal tubular epithelial cells decreased distinctly (P＜0.01), the expression of TGF-betal increased significantly (P＜0.01).Conclusion Repair slows down significantly after kidney ischemia-reperfusion injury and fibrosis occurs gradually in IL-10-/-mice, eventually progressing to chronic kidney disease.

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.

Objective To estimate brain grey matter volume changes and location of abnormal brain regions cerebrum in early stage of bipolar disorder I (BD),in order to provided objective basis for diagnosing early stages BD.Methods 1 7 cases of BD with duration less than 2 years and 1 7 normal controls were recruited in this study.The volumetric difference of grey matter between two groups were analyzed by using voxel-based morphometry(VBM)software.Statistical threshold was voxel> 100,P <0.001 (uncor-rected).Results Compared to the normal controls,the grey matter volume of BD patients decreased in the left dorcial anterior cingu-late cortex(ACC),left insular,right sub-genu ACC,left superior temporal cortex,bilateral hippocampus-parahippocampus-amydala and left posterior lobe of cerebellum(P <0.001).Conclusion The grey matter volume of early stage BD patients is decreased,main-ly locating in the bilateral limbic system,the superior temporal gyrus and the cerebellar cortex,which probably is the morphological appearance of pathomechanism in early stages of BD.

This paper introduces the concepts related to sexual health and sexual health care, summarizes the contents, application scope and limitations of sexual health care assessment tools for oncology nurses at home and abroad, analyzes the problems existing in the assessment tools and puts forward suggestions, aiming at providing theoretical reference for the localization development of sexual health care assessment tools and the development of sexual health care.

Objective To comparatively analyze the safety and efficacy of direct mechanical thrombectomy and bridging therapy for patients with acute anterior circulation large-artery occlusive stroke.Methods A total of 116 patients with acute anterior circulation large-artery occlusive stroke,admitted to our hospitals from October 2015 to March 2018,were chosen in our study;their clinical data were analyzed retrospectively.Among them,63 patients accepted direct mechanical thrombectomy and 53 accepted bridging therapy.The preoperative baseline data and the diagnoses and treatments of the two groups were analyzed;the degrees of modified thrombolysis in cerebral infarction (mTICI),incidences of hemorrhage transformation and symptomatic intracranial hemorrhage,and modified Rankin scale (mRS) scores and mortality rate 90 d after operation were compared between the two groups.Results The preoperative Alberta stroke program early CT scale (ASPECTS) and Glasgow Coma Scale (GCS) scores of the direct mechanical thrombectomy group were significantly lower than those of the bridge therapy group (P＜0.05),and the time from onset to admission was significantly longer than that of the bridging therapy group (P＜0.05).The incidence of postoperative hemorrhage transformation in the direct mechanical thrombectomy group was significantly higher than that in the bridging therapy group (34.9％ vs.17.0％,P＜0.05),but there were no significant differences in the effective recanalization rate (69.8％ vs.79.3％),intracranial symptomatic hemorrhage rate (15.9％ vs.7.6％),favorable outcome rate (28.6％ vs.35.9％) and mortality (22.2％ vs.17.0％) between the two groups (P＞0.05).Conclusion The clinical efficacy and safety of direct mechanical thrombectomy and bridging therapy for patients with acute anterior circulation large-artery occlusive stroke are similar.

Inflammation plays an important role in the development of acute lung injury (ALI). Severe pulmonary inflammation can cause acute respiratory distress syndrome (ARDS) or even death. Expression of proinflammatory interleukin-1β(IL-1β) and inducible nitric oxide synthase (iNOS) in the process of pulmonary inflammation will further exacerbate the severity of ALI. The purpose of this study was to explore the effect of Palrnatine (Pa) on lipopolysaccharide (LPS)-induced mouse ALI and its underlying mechanism. Pa, a natural product, has a wide range of pharmacological activities with the potential to protect against lung injury. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) assays were performed to detect the expression and translation of inflammatory genes and proteins in vitro and in vivo. Immunoprecipitation was used to detect the degree of P65 translocation into the nucleus. We also used molecular modeling to further clarify the mechanism of action. The results showed that Pa pretreatment could significantly inhibit the expression and secretion of the inflammatory cytokine IL-1β, and significantly reduce the protein level of the proinflammatory protease iNOS, in both in vivo and in vitro models induced by LPS. Further mechanism studies showed that Pa could significantly inhibit the activation of the protein kinase B (Akt)/nuclear factor-κB (NF-κB) signaling pathway in the LPS-induced ALI mode and in LPS-induced RAW264.7 cells. Through molecular dynamics simulation, we observed that Pa was bound to the catalytic pocket of Akt and effectively inhibited the biological activity of Akt. These results indicated that Pa significantly relieves LPS-induced ALI by activating the Akt/NF-κB signaling pathway.

Objective:To investigate the mechanism of curcumin affecting HIV-1 infection co-receptor CCR5 by regulating transcription factor FOXP3.Methods:Binding sites of transcription factor FOXP3 on CCR5 promoter were predicted and analyzed by bioinformatics method.AutoDock 4.2 software was used to connect curcumin and FOXP3 flexibly.MTT assay was used to detect cyto-toxcity of curcumin on activity of Jurkat cells.qRT-PCR and Western blot were used to detect expression levels of CCR5 and FOXP3 mRNA and protein in Jurkat cells that were treated with different concentrations of curcumin.pcDNA3.1-FOXP3 expression vector was built and combined with the prediction results of transcription factors.The mutant CCR5 gene fragment was amplified by Overlap PCR,and the mutant CCR5 promoter recombinant vector pFireRluc-Mt-CCR5 was constructed.Binding site between transcription fac-tor FOXP3 and CCR5 promoter was verified by double luciferase reporter gene assay.Results:Results of JASPAR transcription factor prediction showed that there was a binding site between CCR5 promoter and transcription factor FOXP3;molecular docking results showed that curcumin could bind to the active region of FOXP3;MTT results showed that curcumin inhibited the activity of Jurkat cells after 24 hours,and the IC50 was 34.48 μmol/L.qRT-PCR and Western blot showed that expression levels of CCR5 and FOXP3 mRNA and protein were decreased in a dose-dependent manner after different concentrations of curcumin treated Jurkat cells;double luciferase reporter gene confirmed that FOXP3 could bind to CCR5 promoter,and the transcription factor FOXP3 could regulate the activity of CCR5 promoter;results of the recovery experiment of FOXP3 on curcumin showed that when the curcumin concentration was 60 μmol/L,relative value of luciferase activity in HEK293T cells with pcDNA3.1-FOXP3 and pFireRluc-Wt-CCR5 was signifi-cantly higher than that in pFireRluc-Wt-CCR5+curcumin-60 group(P<0.01).Conclusion:FOXP3 can regulate the activity of CCR5 promoter,and the mechanism may be that curcumin affects activity of CCR5 promoter by acting on binding site of FOXP3 and CCR5 promoter.