Background Several cytokines,especially interleukin-1β (IL-β) involve in the breakdown of blood-retina barrier,and the signal of cytokine is transduced through protein tyrosine kinase (PTK) pathway.Objective This study was to investigate the effects of PTK inhibitor,Genistein,on IL-1β-induced blood-retinal barrier breakdown and possible mechanism.Methods The animal models of blood-retinal barrier breakdown were induced through intravitreal injection of IL-1β(10ng) in 24 clean healthy SD rats and assigned to IL-1β group and Genistein group.5μl IL-1β+1μl Genistein with 0.2,1,5μg were intravitreally injected in 12 model rats and 5μl IL-1β (2mg/L)+1μl DMSO was used at the same way in other 12 models.Evans Blue was injected in rats via jugular vein in 1 hour before sacrifice of animals and the arterial blood was collected for the detect of serum Evans Blue.The retinas of the rats were obtained in 4 and 48 hours after injection of vitreous cavity to assay the content of Evans Blue in retina.The changes of vessels and infiltration of inflammatory cells were observed with hematoxylin-eosin stain.RT-PCR was employed to determine the expression of IL-8 and MCP-1mRNA in neuroretina after intravitreal injection.Expression of MCP-1 protein was localized by immunohistochemistry.Results The ratio of retinal Evans Blue and plasma Evans Blue was significantly decreased after intravitreal injection of different doses of Genistein among Genistein groups and IL-8 group with a statistical difference (4 hours:F=7.510,P=0.010;48 hours:F=5.960,P=0.019).With the increase of time after injection of Evans Blue,the ratio of retinal Evans Blue and plasma Evans Blue was gradually reduced in comparison to IL-1β group (P＜0.05).After injection of IL-1β,the dilation of retinal vessel and adhesion of leukocyte to vessel wall were seen under the light microscope,but infiltration of less inflammatory cells was found in Genistein group.The expressions of IL-8 and MCP-1mRNA were obviously declined in retina of rats in Genistein groups compared with IL-8 group (P＜0.05).Immunochemistry indicated that the expression of MCP-1 protein in neuroretina tissue was weaker in Genistein group compared with IL-8 group.Conclusion PTK inhibitor,Genistein,can decrease IL-1β-induced permeability of vessel and maintain the integrity of blood-retinal barrier by downregulating the expression of chemokines and infiltration of leukostasis in retinal vessels.This study imply that PTK pathway plays an important role in IL-1β-induced blood-retinal barrier breakdown.

Objective:To investigate the relationship of etiology and complications of rhabdomyolysis with its prognosis in the elderly.Methods:Patients with rhabdomyolysis at the emergency department of our hospital from January 1, 2018 to December 31, 2019 were retrospectively analyzed.Based on age, patients were divided into the non-elderly group(<65 years old)and the elderly group(≥65 years old). The frequency distribution of etiological factors, concurrent acute kidney injury, and their association with prognosis were analyzed.Results:The number of patients with rhabdomyolysis caused by 2 or more etiologies was higher in the elderly group than in the non-elderly group(40.3% or 48/119 vs.17.0% or 16/94, χ2=13.582, P=0.000). The frequency distribution of etiological factors was different between the two groups.The top-five etiologies were infection, muscle ischemia/hypoxia, endocrine metabolic abnormalities, trauma and muscle fatigue in the elderly group and muscle fatigue, infection, endocrine metabolic abnormalities, drugs/toxicants and trauma in the non-elderly group.Compared with the non-elderly group, the elderly group had fewer patients with typical clinical manifestations(32.8% or 39/119 vs.48.9% or 46/94, χ2=5.067, P=0.024). In contrast, patients who newly presented with disturbance of consciousness were more likely to be found in the elderly group than in the non-elderly group(40.3% or 48/119 vs.21.3% or 20/94)( χ2=7.923, P=0.005). There were 37 patients with AKI(38.9% or 37/95)in the elderly group and 13 of them died(35.1%), and there were 17 patients with AKI in the non-elderly group(19.3% or 17/88)and 4 died(23.5%), indicating the elderly were prone to AKI( χ2=7.545, P=0.006). There was a significant correlation between AKI and prognosis in the non-elderly group( χ2=7.196, P=0.007). Conclusions:Rhabdomyolysis caused by multiple etiologies is more common in elderly patients than in non-elderly patients.The etiological classification of rhabdomyolysis in the elderly is different from that in the non-elderly.Elderly patients are less likely to have typical clinical manifestations and are more prone to AKI.Elderly patients with rhabdomyolysis combined with AKI have a poor prognosis.

Antihypertensive Agents ; therapeutic use ; Endothelin Receptor Antagonists ; therapeutic use ; Familial Primary Pulmonary Hypertension ; Humans ; Hypertension ; drug therapy ; Hypertension, Pulmonary ; drug therapy

Biphasic dissolution test, consisting of immiscible aqueous and organic phase, is an in vitro dissolution method that simultaneously measures the dissolution and partition of drugs. Due to the advantages of simulating in vivo absorption and overcoming the influence of surfactants on dissolution, it has been widely used to evaluate the poorly soluble drugs in vitro dissolution. Based on the relevant research in this field in recent years, this review summarizes the history, dissolution device, theoretical model and application of the biphasic dissolution test. Finally, the prospects in the development of biphasic dissolution test are also outlined.

OBJECTIVETo establish the comparative proteomic profiles of retinoid acid (RA) resistant human acute promyelocytic leukemia (APL) NB4-R1 cells before and after apoptosis induced by realgar (tetra-arsenic tetra-sulfide, As4S4).

METHODSFirst a serial of assays were performed using MTT, transmission electron microscopy, Annexin V FITC/PI double-stain, flow cytometry and confocal laser scanning microscopy to qualitatively and quantitatively observe the in vitro apoptosis inducing effect of realgar on RA-resistant cells. Then the comparative proteomic profile before and after NB4-R1 apoptosis was established using high-resolution two-dimensional electrophoresis system.

RESULTSThe inhibition effect of realgar on NB4-R1 cell growth was dose and time dependent. The 24-h 50% inhibiting concentration (IC50) was 24.06 +/- 0.19 micromol/L, and the 48-h IC50 9.50 +/- 0.13 micromol/L, and 72-h IC50 6.55 +/- 0.03 micromol/L, respectively. 24 h and 48 h were the early and late phase of major NB4-R1 apoptotic cell populations induced by 25 micromol/L realgar respectively. Differential proteomic profiles before and after realgar induced NB4-R1 apoptosis were successfully established. Averagely 1069, 975 and 893 spots could be detected of the untreated group (R0), the 24-h treatment group (R24), and the 48-h treatment group (R48), respectively by ImageMaster 2D Platinum Software. The matching rate between R24 and R0 was 79.94% and that between R48 and R0 69.33%, and that between R24 and R48 71.91%.

CONCLUSIONDifferential proteomic profiles of realgar induced NB4-R1 apoptosis were successfully established for the first time, which provided a basis for comprehensively understanding the signal transduction of realgar induced apoptosis in RA-resistant APL cells, also for screening new bio-markers and drug targets of hematopoietic malignant tumor.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; drug effects ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Proteome ; analysis ; Sulfides ; pharmacology ; Tretinoin ; pharmacology

ObjectiveTo compare the intraoperative wake-up test in patients undergoing scoliosis surgery with different anesthesia methods.MethodsForty ASA Ⅰ patients aged 13-18 yr with body mass index ＜ 30 kg/m2 scheduled for scoliosis surgery were randomly divided into 2 groups ( n =20 each): propofol combined sufentanil anesthesia group (group P) and sevoflurane combined sufentanil anesthesia group (group S).Anesthesia was induced with target-controlled infusion of sufentanil(target effect-site concentration 0.5 ng/ml),and iv injection of etomidate 0.3 mg/kg in both groups.Tracheal intubation was facilitated with 0.15 mg/kg cisatracurium when patients lost consciousness.The patients were mechanically ventilated.Anesthesia was maintained with target-controlled inhalation of sevoflurane (target end-tidal concentration 0.8％-1.5％ ) in group S,and target-controlled infusion of propofol (target plasma concentration 3-5 μg/ml) in group P,and target-controlled infusion of sufentanil (target effect-site concentration 0.2-0.3 ng/ml),and iv infusion of cisatracurium 0.1 mg· kg-1· h-1 in both groups.BIS value was maintained at 40-60.Cisatracurium administration was terminated and target effect-site concentration of sufentanil decreased to 0.1 ng/ml before wake-up test,5 min later,sevoflurane and propofol administration were terminated,and 5 min later wake-up test was performed.MAP and HR were recoreded during wake-up test.The wake-up time and advers effect (bucking,restlessness and awareness)were recorded.Results The wake-up time was significantly shorter in group S than in group P( P ＜ 0.05).MAP and HR were in normal range during wake-up test in both groups,and bucking,restlessness and awareness were not found in both groups.ConclusionTarget-controlled inhalation of ssvoflurane combined with sufentanil can be safely and effectively used for intraoperative wake-up test in patients undergoing scoliosis surgery,and the wake-up time is shorter than that with propofol combined sufentanil,and it is an apporiate anesthetic technique for the intraoperative wake-up test.

Objective To investigate the neuroprotective effects of neurotrophin-3 (NT-3) expression controlled by five copies of the hypoxia-responsive elements after focal cerebral ischemia .Methods Three groups of rats re-ceived RV-5H-NT3, RV-5H-EGFP or saline injection .Three days after gene transfer , the rats underwent 90 min of transient middle cerebral artery occlusion ( tMCAO) , followed by 1-28 days of reperfusion .Immunohistostaining and western blotting were performed to detect ischemia/hypoxia-regulated expression of NT-3 controlled by HRE . The volume of brain infarction and the apoptosis were analysised by TTC and TUNEL staining .The neurological scoring was determined by neurological behavior tests .Results Three days after tMCAO , brain NT-3 expression was significantly increased in the RV-5HNT3-transduced animals compared with the RV-5H-EGFP or saline group (P<0.05), and brain infarct volume was smaller in the RV-5H-NT3-transduced group than the RV-5H-EGFP or saline group ( P<0.05 ) .The percentage of TUNEL-positive cells was reduced in RV-5 H-NT3-transduced brains compared with the RV-5 HEGFP or saline group 3 and 7 days after tMCAO ( P<0.05 ) .Furthermore , the neurolog-ical status of RV-5H-NT3-transduced rats was better than that of RV-5H-EGFP-or saline-transduced animals from 1 day to 4 weeks after tMCAO ( P<0.05 ) .Conclusions HRE may modulate NT-3 expression in the ischemic brain tissue and that the up-regulated NT-3 may effectively improve neurological status following tMCAO due to de-creased initial damage .

Objective To construct and identify the recombinant retroviral vector containing five copies of hypoxia responsive elements (5HRE)and neurotrophin-3 (NT-3 ).Methods Using PCR,enzyme digestion and DNA ligase,5HRE and human derived NT-3 were cloned into the retroviral vector plasmid (pLNCX)to construct the recombinant retroviral vector plasmid pLNCX-5HRE-SV40-NT3-IRES-EGFP.The retrovirus RV-5HRE-NT3 was packaged in the PT67 cells,and then it was purified and concentrated by high-speed centrifugation.After infected for 48 h with the concentrated retrovirus,the number of the EGFP positive cells in the NIH 3T3 cells was counted by fluorescence activated cells and sorted to calculate the retrovirus titer.Results The retroviral vector plasmid,pLNCX-5HRE-SV40-NT3-IRES-EGFP,was successfully constructed,and the retrovirus was packaged and defined as RV-5HRE-NT3.After purification and concentration,the retrovirus titer reached 9.1 × 10 6 cfu/mL. Conclusion The recombinant retroviral vector which carried out hypoxia-regulated expression of NT-3 was successfully constructed.It may provide basis for studies on hypoxia-regulated expression of the exogenous genes.

This study aimed at unfolding the therapeutic effects of freeze-dried powder separated from Shuang Xia decoction (SXD) on female senile drosophila melanogaster with fragmented sleep.Taking drosophilas as the model organisms,the locomotor activity was monitored using the autonomic monitoring software to explore the regulation of fragmented sleep in senile drosophilas by the treatment of SXD.As a result,it was found that the optimum concentration of SXD was 2.50％,while the desirable therapeutic duration was 4 days.In comparison with the control group,35-day-old virgin drosophilas'sleep was prolonged in the SXD group and the positive control group.The positive drug mainly affected their sleep in the daytime,while SXD impacted their sleep at night featuring the prolonged fragmented sleep.In conclusion,it was demonstrated that the freeze-dried powder of SXD effectively alleviated intermittent insomnia in the female senile drosophilas.Compared with positive drug,SXD also mitigated intermittent insomnia at night without significant changes in the sleep of the drosophilas in the daytime.Above all,SXD was beneficial in the regulation of sleep-wake rhythm in the drosophilas.

OBJECTIVE The purpose of this study is to clarify the association between Cu levels and heart failure(HF)using a meta- analysis approach. METHODS We searched articles in the PubMed, EMbase, CNKI, Wanfang ,VIP and CBM Database published as of August 2016. The case control study on the relationship between serum copper levels and HF were collected and read and extracted by two independent researchers. A Meta analysis was conducted using Stata 12.0 software. RESULTS A total of twenty- one eligible articles, including 893 HF and 654 control subjects, were enrolled. The Meta analysis showed that serum copper levels in HF were higher than control group〔SMD=0.881, 95%CI: (0.487 ,1.264), Z=4.5, P<0.001〕. The sensitivity analysis indicated that the results were reliable. Begg's tests did not find the existence of publication bias. CONCLUSION This meta-analysis indicates that there is a significant association between high Cu serum level and HF.