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 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.

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 investigate ht e clinical characteristics and treatment of children with acute laryn gitis comlp icated with negative pressure pulmno ary edema(NPPE),and the changes of inflammatory factosr w ere monitored.Methods Data of 9 cases with acute laryngitis complicated with NPPE in pediatric intensive care unit from August 2010 to March 2015 we re analyzde .The levelso f TNF -αand IL-6 of 8 cases were detected at admission and checked agani forty-eihg t horu s after therapy.Ten children of acute laryngitis wi thotu NPPE were selected as disease cotn rol group, and ten healthy children as normal control group. Results (1)The onset of NPPE varied from 8 minutes to 2 hours following relief of obstruction,and presen-ted with acute respiratory disrt ess, decreased xo ygen saturation, tachycardai , rales on chest auscultation.All thees patients received therapeutic measures icn luding mechna ical ventilation,limiting the fluid input volume. The disappearance of rales on chest auscultation varied from 6 hours to 30 hours.Duration of mechanical ven-tilation was lse s than 48 hours,and all the children were cured.(2) Compared with the children of disease control group and normal control group,in acute phase the plasma levels fo TNF-αand IL-6 in children with NPPE were significantly higher ( P<0.01 ) .The indicators of NPPE group significantly decreased after 48 hours therapy( P<0.01 ) .Conclusion NPPE is manifested by rapid onset of respiratory distress after relief of the airway obstruction.The symptoms resolve rapidly if early support of breath and limiting the fluid input volume are applied properly.The inflammatory response is one of the possible mechanisms of NPPE.

Objective To investigate the expression of Toll-like receptor 4 ( TLR4 ) and nuclear factor-κB ( NF-κB) in patients with critically severe hand, foot and mouth disease ( HFMD) and to evaluate the effects of esmolol intervention on those patients.Methods Fifty-two hospitalized children with critically severe HFMD in the Intensive Care Unit of Xuzhou Children′s Hospital were enrolled in the study from May 2014 to May 2015 and randomly divided into two groups, represented as group A and group B.Children in the group A were given routine treatment, while those in the group B were treated with esmolol in addition to the routine therapy.Thirty children with common HFMD were selected as disease control, and thirty healthy children were set up as normal control.Differences in the expression of TLR4, NF-κB, TNF-αand IL-6 among all children were comparatively analyzed.The levels of TLR4, NF-κB, TNF-αand IL-6 in children from groups A and B were detected after 24 hours, 72 hours and five days of treatment and the differences betweenthetwogroupswereanalyzed.Results (1)Comparedwiththechildrenfromdiseasecontroland normal control groups, those with critically severe HFMD showed significantly increased expression of TLR4, NF-κB, TNF-αand IL-6 (P<0.01).(2)No significant differences in the expression of TLR4, NF-κB, TNF-αand IL-6 were found between the two subgroups of children with critically severe HFMD before treat-ment (all P>0.05).The expression of TLR4, NF-κB, TNF-αand IL-6 in children from both subgroups were significantly decreased after receiving corresponding treatments for 24 hours, 72 hours and five days (all P<0.05).Compared with the children form group A, those from group B showed significantly decreased expression of NF-κB, TNF-αand IL-6 after 24 and 72 hours of corresponding treatments (all P<0.05).No significant difference in the expression of TLR4 was observed between the two subgroups after 24 and 72 hours of corresponding treatments (P>0.05).No significant differences in those observed indicators were found between the two subgroups after five days of treatments (all P>0.05).Conclusion The TLR4/NF-κB/proinflammatory factor pathway might play an important role in the development of critically severe HFMD.Treatment with esmolol could inhibit the expression of NF-κB, reduce the secretion of inflammatory factors and alleviate the inflammatory reaction during critically severe HFMD.