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Objective To investigate the clinical features and the clinical effects of extracorporeal membrane oxygenation(ECMO) in children with fulminant myocarditis. Methods The data of one hundred and sixty one pediatric patients with fulminant myocarditis at Beijing Children′s Hospital from March 1,2007 to May 31,2018 were analyzed retrospectively,including clinical manifestations,electrocardiogram,echocar￣diography,myocardial enzyme,the application of ECMO,curative effect and prognosis. Results Among 161 cases of children with fulminant myocarditis,74 cases (46. 0%) had respiratory tract symptoms,54 cases (33. 5%) had digestive tract symptoms,24 cases (14. 9%) had nervous system symptoms,only 33 cases (20. 5%) had cardiovascular symptoms. One hundred and thirty￣two cases(82. 0%) had elevation of crea￣tine kinase isoenzymes( CK￣MB) and/or cardiac troponin I( cTnI),and 107 cases(66. 5%) had abnormal echocardiography. One hundred and fifty￣two cases ( 94. 4%) showed abnormal electrocardiogram, and the changes of ST￣T and atrioventricular block were most common. Five cases survived in 8 children treated with ECMO. They underwent ECMO with an median supporting time of 101. 5 (52. 75,142. 8) hours through a venous￣arterial ECMO model. After the application of ECMO,blood pressure,the heart rate and oxygen satura￣tion were obviously improved in the 8 children,and the level of blood lactic acid was statistically reduced com￣pared with that before the application of ECMO(P<0. 001). During the follow￣up period of 1 months,the 5 surviving children recovered with normal cardiac function,without abnormal function of other organs. There were a total of 17 cases(10. 6%) died,4 children(8. 9%) in 45 patients died since the application of ECMO and the other 13 children(11. 6%) in 116 patients died before that time. This proportion of death was reduced after the application of ECMO,whereas the difference was not statistically significant(P>0. 05). Conclusion Our study indicates the diverse symptoms in children with fulminant myocarditis. Comprehensive application of electrocardiogram,echocardiography,and myocardial enzymology markers may be of benefit to the timely diag￣nosis of fulminant myocarditis. The application of ECMO in fulminant myocarditis remains an effective approach in reduction of mortality rate,and can effectively support recovery of cardiopulmonary functions.
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Objective To evaluate the role of hippocampal neuronal nitric oxide synthase(nNOS)-postsynaptic dense protein 95(PSD95)coupling in short-term memory retrieval disorder induced by sevoflu-rane in mice.Methods Sixteen clean-grade healthy Kunming mice of both sexes,aged 2-3 months,weighing 30-35 g,were divided into 2 groups(n＝8 each)according to the random number table method:sevoflurane group(S group)and nNOS-PSD95 uncoupling agent ZL006 group(Z group).After successful establishment of dark avoidance memory,3.3％sevoflurane and 40％O2 were inhaled for 2 h in both groups,and normal saline 1.5 ml was intraperitoneally injected in group S and ZL006 1 mg/kg in group Z at 30 min before anesthesia.The step-through latency and error times were recorded before anesthesia and at 12 h after the end of anesthesia.The mice were then sacrificed,and hippocampal tissues were taken for de-termination of the expression of nNOS and PSD95(by Western blot)and co-expression of nNOS and PSD95(by immunoprecipitation).Results Compared with that before anesthesia,the step-through latency was significantly shortened,and the error times were increased at 12 h after anesthesia in group S(P＜0.05),and no significant change was found in the above indicators in group Z(P>0.05).Compared with group S,the step-through latency was significantly prolonged,error times were decreased,the co-expression of nNOS and PSD95 was down-regulated(P＜0.05),and no significant change was found in the expression of nNOS and PSD95 in group Z(P>0.05).Conclusion The mechanism by which sevoflurane induces short-term memory retrieval disorder may be related to promoting the coupling of nNOS to PSD95 in the hippocam-pus of mice.
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Aim To detect the effects of cetuximab combined with adriamycin on the proliferation and ap-optosis of triple-negative breast cancer cells.Methods Cell viability was evaluated by MTT assay.The cell apoptosis was analyzed by flow cytometry with propidi-um iodide staining.JC-1 staining was used to deter-mine mitochondrial membrane potential.The expres-sions of glucose regulated protein78 (GRP-78),Bcl-2 and Caspase-3 were measured with Western blot.Re-sults MTT assay showed that cetuximab had inhibi-tion effect on the breast cancer cell MDA-MB-231 growth,and the effect was related to concentration of drug.The inhibition effect of adriamycin on MDA-MB-231 had remarkabe relationship with time and concen-tration.When combined with each other,they could re-markably increase inhibition effect.The viability of cells in combination group for 1 2 h,24 h,48 h,sig-nificantly lower than that in cetuximab or adriamycin group (P group (P <0.01 ).JC-1 staining indicated that cetux-imab or adriamycin could reduce the mitochondrial membrane potential,but the reduction effect was more remarkable in the combination group.Western blot re-vealed that cetuximab could reduce the expression of GRP-78 and Bcl-2,and increased the expression of Caspase-3 and its activity.The expressions of Bcl-2, Caspase-3 had no significant change in adriamycin group,but GRP-78 was increased.In combination group,the expression of GRP-78 and Bcl-2 was signifi-cantly decreased,but Caspase-3 was increased nota-blely compared to adriamycin group.Conclusions The combination of cetuximab and adriamycin enhances the inhibition effect on the triple-negative breast cancer MDA-MB-231 cells,and increases cell apoptosis.The mechanism may be that cetuximab reduces the endo-plasmic reticulum stress level,then activates the mito-chondrial pathways by decreasing the expression of Bcl-2,reducing the mitochondrial membrane potential,and promoting cell apoptosis.
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Aim To investigate the anti-proliferative effect of salinomycin on doxorubicin-resistant human breast cancer MCF-7 /DOX cells.Methods MCF-7 and MCF-7 /DOX cells were treated or untreated with salinomycin.Cell viability was detected by MTS assay. Cell apoptosis was detected by Annexin V-FITC /PI as-say.Reactive oxygen species (ROS)was measured by DCFH-DA staining.Mitochondrial membrane potential was measured by JC-1 assay.The expression of apopto-sis related proteins BAX, BCL-2, caspase-3, and caspase-9 were evaluated by Western blot analysis. Results The cell viability was significantly reduced by salinomycin treatment in a dose-dependent manner. The flow cytometry results showed that salinomycin in-duced MCF-7 /DOX cell apoptosis,increased ROS pro-duction,and decreased mitochondrial membrane poten-tial.Furthermore,salinomycin decreased the expres-sion of BCL-2,and increased the expression of BAX, cleaved caspase-3,and cleaved caspase-9.Moreover, the antioxidant N-acetylcysteine (NAC ) markedly blocked the above effects.Conclusions Our results suggest that salinomycin-induced apoptosis in MCF-7 /DOX is associated with induction of ROS production, and activation of mitochondria apoptosis pathway, which may become a potential chemotherapeutic agent for the therapy of doxorubicin resistant breast cancer.
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Objective To construct an efficient eukaryotic expression recombinant vector of human interleukin-1O(hIL-lO),and observe its expression in rabbit synoviocytes(RSCs).Methods Total RNA was extracted from peripheral blood mononuclear cells(PBMCs)of a patient with drug allergy.Specific Drimers for full-length open reading frames(ORFs)of hIL-10 were designed according to GeneBank(NM 000572).Withtotal RNA as the template,full-length ORFs of hIL-10 were amplified by reverse transcription Dolymerase chain reaction(RT-PCR).RT-PCR products were digested by restrictive endonucleotidase.then inserted into plasmid pcDNA4/HisMaxA.Both restrictive endonucleotidase analysis and DNA sequencing Were carried out for inserts verification.RSCs were transfected with recombinant plasmid expression vector PcDNA4 HisMaxA-hiL10 by liposome-mediated gene transfer methods,then cultured in vitro.The supernatants were collected af-ter transfection for 12 hours,24 hours,48 hours,72 hours,7 days,14 days respectively for IL-10 measure-ment by enzyme linked immunosorbent assay(ELISA).Results Full-length ORFs of hIL-1o(0.54 kb)had been successfully cloned from PBMCs through RT-PCR.The inserts and insert location of pcDNA4 HisMaxA were in a fight way verified by enzyme analysis and DNA sequencing.ELISA results showed that exogenous hIL-10 gene had expressed in the transfected RSCs from 12 hours to 7 days after transfection,and hIL-10level of transfection group significantly higher than that of the control group.Conclusion pcDNA4 HisMaxA-hiL10,the hIL-10 efficient eukaryotic expression vectors,has been suecessfully constructed.
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Objective To display of heterologous proteins on the surface of E. coli . Methods The 1653 bp luciferase report gene was knocked in Ipp and ompA genes of chromosome by Red recombine system and selection-counter selection kan/sacB. Results The quantitative analysis results of exogenous lu-ciferase expression displayed that it could be expressed as fusion with the outer membrane proteins on the cell surface. The fusion protein of foreign protein and outer membrane protein Lpp-OmpA-Luc could be high-effi-ciently displayed on cell surface. Conclusion The stable expression of exogenous gene on the surface of E. coli had no effect on the bacterial growth and propagation.
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OBJECTIVE: It has been proposed that the contact between blood and dialysis membrane during hemodialysis therapy induces harmful reactions to patients. Membrane biocompatibility is the characteristic that makes less adverse reaction. We tried to compare the biocompatibility between Cuprophane and Polysulfone membranes. MRTHODS: Nine chronic renal failure patients who have undergone maintenance hemrodialysis therapy with Hemophan membrane three times per week were included in this study. Cuprophane membranes were used in the first week: Hemophan membranes in the second week; Polysulfone membranes in the third week. Each membrane was used three times a week. On the day of third dialysis with the Cuprophane membrane(first week) and Polysulfone membrane(third week), serial blood sampling was obtained from the afferent line at hemodialysis initiation, 15 minutes, 30 minutes, 60 minutes, 120 minutes and 30 minutes to measure serum complement activity(C3a, C5a), blood polymorphonuclear leukocyte and platelet count, and arterial oxygen pressure which have been regarded as biocompatibility parameters. The parameters measured during the first week(Cuprophane) and the third week(Polysulfone) were compared to evaluate the difference in biocompatibility of both membranes. RESULTS: 1) C3a desArg In both groups, the level of C3a desArg increased significantly from basal level and the Cuprophane group showed significantly higher level of C3a desArg than that of Polysulfone group. 2) C5a desArg : In both groups, the level of C5a desArg did not increased sigpificantly from basal level. Only at 30 minute after hemodialysis, Cuprophane group showed significantly higher level of C5a desArg than that of Polysulfone group(p=0.037). 3) Polymorphonuclear leukocyte : In both groups, the polymorphonuclear leukocyte counts decreased significantly at 15 minutes and 30 minutes from basal level. The polymorphonuclear leukocyte count was lower in Cuprophane group than that of Polysulfone group at 15 minute after hemodialysis(p=0.001). 4) Platelet: In Cuprophane group, the platelet count was decreased significantly at 15 minute(p=0.004) but there were no difference in platelet counts between the two groups. 5) Arterial oxygen pressure Both group showed no consistent pattern of variation of oxygen pressure and there was no significant difference between the two groups. CONCLUSION: The biocompatibility of Polysulfone membrane was superior to the Cuprophane membrane with respect to the increased activation of complement C3a. decrease of polymorphonuclear leukocyte and decrease of platelet count.
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Humans , Blood Platelets , Complement C3a , Complement System Proteins , Dialysis , Kidney Failure, Chronic , Membranes , Neutrophils , Oxygen , Platelet Count , Renal DialysisABSTRACT
Objective: Effects of Shenmai injection and aminophylline alone or in combination on transdiaphragmatic pressure and electrical activity of diaphragm were observed, so as to evaluate their effecfs on diaphragmatic fatigue. Methods: The diaphramatic fatique (DiF) model was estab- lished by stimulating bilateral phrenic nerves for 40 minutes. The animals were allocated to three groups: Shenmai group (2mL/kg, N=7), aminophylline group (20mg/kg, N=7) and combi- nation group (Shenmai injection 2mL/kg+aminophylline 20mg/kg, N=7) The transdi- aphragmatic pressure (Pdi) and diaphragmatic electromyogram (EMGdi) were recorded before and after treatment. EMGdi was analysed by computer and then the high/low frequency ratio (H/L) and the central frequency (Fc) were caculated. The diaphragm evoked potential (DEP) was record- ed simutaneously. All data was analysed by analysis of variance and q test. Results: Pdi, H/L, Fc and amplitude of DEP were markedly increased after the treatment with aminophyline and Shen- mai injection alone (Compared with DiF. P