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【Objective】 To establish a novel preparation method of cryoprecipitate coagulation factor from overcooled liquid-state plasma. 【Methods】 The fresh liquid plasma was kept at -11℃ to -13℃ for a period of time. It can remain in the liquid state with some coagulation factors generated due to supercooling. Then cryoprecipitate can be obtained from the liquid plasma by siphon method. 【Results】 The average fibrinogen content yielded in cryoprecipitate, prepared from 50 samples of 16-hour-stored fresh liquid plasma, was (186.02±22.72) mg, with the average recovery rate of (37.51±7.42) %, and the average content of coagulation FⅧ was (104.66±22.88) IU, with the average recovery rate of (46.62±5.58) %. 【Conclusion】 The cryoprecipitate coagulation factors could be obtained not only from fresh frozen-thawed plasma, but also from overcooled liquid plasma which is simple and stable, also meets the requirements of relative standards.
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To study the interaction between C4b-binding protein (C4BP) and Riemerella anatipestifer (RA), we cloned duck C4BPα, conducted prokaryotic expression and prepared the polyclonal antibody by immunizing mice. Then indirect immunofluorescence assay and dot blotting hybridization assay were used to verify the interaction between C4BP and RA. The full length of duck C4BPα nucleotide sequence was 1 230 bp, with the highest similarity to chicken C4BPα (82.1%). Phylogenetic tree analysis showed that duck C4BPα and chicken C4BPα were on the same phylogenetic tree branch and the genetic evolution relationship between them was the closest. C4BPα was efficiently expressed in Escherichia coli BL21 (DE3). The recombinant proteins existed in intracellular soluble form. The titer of polyclonal antibody was more than 1:10 000 and polyclonal antibodies could specifically recognize the recombinant proteins. The results of indirect immunofluorescence assay and dot blot hybridization assay showed that RA could interact with duck C4BP. The results provide a basis to further reveal the pathogenesis of RA.
Subject(s)
Animals , Mice , Cloning, Molecular , Complement C4b-Binding Protein , Chemistry , Genetics , Metabolism , Ducks , Classification , Genetics , Microbiology , Gene Expression Regulation , Phylogeny , Riemerella , MetabolismABSTRACT
Objective TosummarizeMRIfeaturesofadultreversiblespleniallesionsyndrome(RESLES)andevaluateitsdiagnosticvalue. Methods ClinicalandMRIdataof15patientswithRESLESwereanalyzedretrospectively.Primarydiseasesandcausesincluded:encephalitis disease,cerebrovasculardiseases,metabolicandelectrolytedisorders.Theclinicalsymptomswereheadache,dizziness,changeofconsciousness, epilepticseizuresandothernonspecificneurologicsymptoms.Thecranialimagingexaminationwasperformedat3.0T/1.5TsuperconductingMR, includingT1WI,T2WI,T2FLAIRandDWI,inwhich7casesunderwentMRIenhancedscanand5casesunderwentMRAexamination.Results Among15patients,MRIfindingsofallpatientswerecharacterizedbythecharacteristic"boomerangsign"ortheovoidabnormalitysignalinthe corporiscallosisplenium,whichwaspresentedascytotoxicedema.Besides,4ofthem werecombinedwithposteriorreversibleencephalopathy syndrome(PRES),whichwascharacterizedbyvasogenicedemainthebilateraloccipital,temporal,parietallobeandcerebellarhemisphere.The clinicalsymptomsofallpatientsimprovedafterreasonabletreatment.Conclusion MRIcharacteristicfindingofadultreversiblesple-niallesionsyndromeisthecytotoxicedemaofthecorpuscallosum,whichcanbeshowedas"boomerangsign"orovalform.Imagingfindingsand clinicalsymptomsarereversible.Thecorrectunderstandingofthisdiseaseisofgreatsignificancetoclinicalrationaldiagnosisandtreatment.
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Objective To explore the relevance between nitrate transporters (Sialin) and ischemia-reperfusion injury (IRI) in free flaps.Methods Twenty rats were randomly divided into control group and experimental group,10 in each group.After preparatiing 4 cm×8 cm free tissue flap on left lower abdominal,the rats in experimental group were experienced ischemia in 6 hours,then reperfusion.The rats in control group were without ischemia treatment,and tissue flaps were sutured in situ.After postoperative observation for 7 days,the furthest edge of survival flaps were sampled.The expression of Sialin was detected by the Real-time PCR and Western blot.The change of nitrate content was analysed by chemiluminescence.Results Both of results of Real-time quantitative PCR and Western blot,it was found that expression levels of Sialin in the experimental group were significantly lower than the control group(0.945±0.0530)(P＜0.05).Applying Real-time quantitative PCR relative quantitative analysis,it was found that the experimental group Sialin mRNA expression level was 0.284±0.0486,significantly lower than that of control group (0.945±0.0530)(P＜0.05).Using Western blot technical analysis,experimental Sialin protein expression level was(0.1413±0.0446),significantly lower than the control group (0.3519±0.0368) (P＜0.05).The concentration of nitrate of test results found that the expression of nitrate transporters (Sialin) linearly related with the concentration of nitrate in free flaps,which means tbe Sialin expression level decreased,the nitrate content decreased(P＜0.05,R=0.81).Conclusion The concentration of nitrate was linearly related to the level of Sialin expression:when Sialin expression levels dropped,the concentration of nitrate decreased.Nitrate transporters(Sialin) may be correlated with IRI in free flaps.
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Objective To evaluate the effects of dexmedetomidine on the expression of tumor necrosis factor-α(TNF-α) and intercellular adhesion molecule-1 (ICAM-1 ) during cerebral ischemia-reperfusion (I/R ) in rats .Methods Thirty-six male Sprague-Dawley rats , aged 8-12 weeks ,weighing 280-320 g ,were randomly divided into 3 groups ( n= 12 each ):sham operation group (group S ) , cerebral I/R group (group I/R ) and dexmedetomidine group (group D ) .The animals were anesthetized with 10% chloral hydrate 3.5 ml/kg .The focal cerebral ischemia was induced by middle cerebral artery occlusion using a nylon thread with rounded tip inserted into internal carotid artery and advanced cranially until resistance was met .In group D ,dexmedetomidine 3μg/kg was injected intravenously immediately after beginning of ischemia ,followed by infusion at 3μg·kg-1 ·h-1 for 120 min ,while the equal volume of normal saline was given in group I/R .Four rats from each group were chosen at 24 ,48 and 72 h of reperfusion , and blood samples were taken from the left ventricle to determine the concentrations of serum S100B protein .The rats were then sacrificed and the brains were removed to determine the expression of TNF-αand ICAM-1 in brain tissues .Results Compared with group S ,the concentrations of serum S100B protein were significantly increased , and the expression of TNF-α and ICAM-1 in brain tissues was up-regulated in I/R and D groups ( P<0.05) .Compared with group I/R ,the concentrations of serum S100B protein were significantly decreased , and the expression of TNF-α and ICAM-1 in brain tissues was down-regulated in group D ( P<0.05 ) .Conclusion The mechanism by which dexmedetomidine reduces the cerebral I/R injury is related to down-regulation of TNF-αand ICAM-1 expression in rats .
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Objective To investigate the value of plasma miR-22 in diagnosis of idiopathic pulmonary arterial hypertension ( IPAH ),and its role of regulation mechanisms in the pathogenesis of the disease.Methods Circulating miR-22 levels of IPAH patients and healthy controls were evaluated by RTPCR.The silico analysis of targets for miR-22 was taken, and followed by eGFP reporter assay for verification of predicted target gene Myc binding protein (MYCBP). Results Compared with healthy controls,the expression of plasma miR-22 in IPAH patients was significantly decreased (P ＜ 0.01 ).The area under curve (AUC) of ROC curve was 0.744.MYCBP was a real target of miR-22 confirmed by silico analysis and eGFP reporter assay. Conclusions The expression of plasma miR-22 was significantly decreased in IPAH patients,and it could serve as a potential biomarker for diagnosis.The miR-22 might be involved in the pathogenesis of the disease through promoting its target gene MYCBP to activate the c-Myc pathway.
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Objective To investigate the effect of preconditioning with different concentrations of sevoflurane on hypoxia-reoxygenation(H/R)-induced apoptosis in rat hippocampal neurons and the role of mitochondrial KATP(mito-KATP)channels.Methods Primary cultured hippocampal neurons isolated from newborn SD rats(＜24h)of both sexes,weighing 5-6 g,were randomly divided into 7 groups with 48 wells and 12 dishes in each one:control group(C group),H/R group,preconditioning with 6%,4%and 2% sevoflurane groups(S1-3 groups),5-hydroxydecanoate(5-HD,mito-KATP channel blocker)100 μmol/L preconditioning group(5-HD group)and preconditioning with 5-HD 100 μmol/L+6% sevoflurane group(5-HD+S group).The neurons were exposed to 4 h hypoxia followed by 24 h reoxygenation. In S1-3 groups, preconditioning was performed with 6% , 4% and 2% sevoflurane respectively before H/R. In 5-HD group, preconditioning was performed with 5-HD (final concentration 100 μmol/L) before H/R. In 5-HD + S group, preconditioning was performed with 5-HD 100 μmol/L and 6% sevoflurane before H/R. The neuronal viability, apoptosis rate and expression of Bcl-2 and Bax were determined after 24 h reoxygenation.Results The neuronal viability was significantly lower,while the apoptosis rate and expression of Bcl-2 and Bax were significantly higher in the other 6 groups than in group C(P＜0.01).The neuronal viability and expression of Bcl-2 were significantly higher,while the apoptosis rate and Bax expression were lower in S1-3 groups than in group H/R. There was no significant difference in the parameters mentioned above between 5-HD and 5-HD + S groups(P＞0.05).The neuronal viability and expression of Bcl-2 were significantly lower, while the apoptosis rate and Bax expression were higher in S2, S3 and 5-HD + S groups than in group S1, and in group S3 than in group S2(P＜0.0l) .Conclusion Sevoflurane preconditioning can inhibit H/R-induced apoptosis in rat hippocampal neurons and reduce the injury to neurons in a concentration-dependent manner, and the underlying mechanism may be related to activation of mito-KATP channels, up-regulation of Bcl-2 expression and down-regulation of Bax expression.
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@# Objective To observe the effect of the soft tissues injury around atlantoaxial joint on memory abilities of mice.Methods 44 mice were divided into the model group and control group with 22 animals in each group.The soft tissues around atlantoaxial joint were injured in the model group by surgery between the first and the second cervical vertebrae.The changes of learning and memory of mice were observed with step-down avoidance test,water-maze test and hole-board test;the expression of caspase-3 in hippocampus was observed by immunohistochemical staining.Results Compared with the control group,escape latency was longer and wrong times were more in water-maze test of the model group(P<0.01). In step-down avoidance test of the model group,the reaction period prolonged and wrong times increased obviously(P<0.01).In hole-board test,the latency period prolonged and the times of extending the holes decreased significantly(P<0.01).Degenerated or dead neuron in hippocampus of surgery-treated mice was found scatteredly or continuously with HE staining,the expression of caspase-3 protein was showed positive with immunohistochemical staining.Conclusion The soft tissues injury arround atlantoaxial joint can induce abnormality of the stability of cervical vertebra,which aggravated the aging process of the mice and decreased abilities of learning and memory.