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Objective:To investigate the protective effect of lipopolysaccharide (LPS) preconditioning on cerebral ischemia reperfusion in rats.Methods:One hundred and twenty adult male SD rats were randomly divided into sham operation group, model group, low-dose LPS group (0.05 mg/kg), medium-dose LPS group (0.15 mg/kg), and high-dose LPS group (0.45 mg/kg). LPS was injected intraperitoneally for preconditioning, once a day for 7 consecutive days. Twenty-four hours after the last injection, the left middle cerebral artery occlusion model was induced by suture-occluded method. The model was reperfused 1.5 h after ischemia. At 24 h after reperfusion, the neurological deficit was evaluated by neurobehavioral score. The volume of cerebral infarction was measured by triphenyltetrazolium chloride staining. The serum levels of proinflammatory cytokines interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay. The expression of Toll-like receptor 4 (TLR4), matrix metalloproteinase (MMP) -2 and MMP-9 in ischemic brain tissue was detected by Western blot analysis.Results:Compared with the sham operation group, blood-brain barrier permeability was increased in the model group, serum IL-1β, IL-6 and TNF-α were up-regulated, and the expression of TLR4, MMP-2 and MMP-9 proteins in ischemic brain tissue was up-regulated. Compared with the model group, the neurological impairment of each LPS intervention group was significantly reduced, the volume of cerebral infarction was significantly reduced, the permeability of blood-brain barrier was significantly reduced, the serum IL-1β, IL-6 and TNF-α were down-regulated, and the expression of TLR4, MMP-2 and MMP-9 protein in ischemic brain tissue was down-regulated, especially in the medium-dose group and the high-dose group.Conclusions:LPS preconditioning can induce the formation of ischemic tolerance, inhibit the activation of TLR4-MMP-2/MMP-9 signal pathway, reduce the damage and inflammation of blood-brain barrier structure, and thus play a neuroprotective role in rats with cerebral ischemia reperfusion.
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@#Objective To investigate the role of miR-132 in oxygen-glucose-deprivation （OGD） induced ischemic injury model using primary cortical neurons.Methods Cultured primary cortical neurons were exposed to oxygen-glucose-deprivation （OGD）for 2 h followed by 24 h reperfusion.The cells were randomly divided into sham operation group (Sham),OGD group,lentivirus control group (LV-control),miR-132 low expression group (LV-anti-miR-132),and miR-132 overexpression group (LV-miR-132).CCK-8,real-time PCR,and Western blot were used to study the effects and mechanisms of miR-132 on primary cortical neurons injured by OGD.Results The expression levels of miR-132 were significantly reduced in the OGD-induced ischemic injury model (P<0.05).The reduced expression of miR-132 aggravated OGD injury and significantly reduced cell viability in OGD-induced primary cortical neuron ischemic injury.On the contrary,the increased expression levels of miR-132 reduced OGD injury and increased cell survival.Western blot results showed that overexpression of miR-132 upregulated the levels of synapsin-1 and PSD95.Conclusion Overexpression of miR-132 can improve OGD-induced ischemic injury and increase the survival of primary cortical neurons by upregulating the synapsin-1 and PSD95.
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Objective To explore the effect of group cognitive behavior intervention on self-rated health of middle school students with emotional disorders. Methods From January 2018 to June 2018,79 middle school students with emotional disorders were randomly divided into intervention group ( 41 cases) and control group (38 cases) according to the single or double number of medical records. The control group only received drug treatment,while the intervention group received group cognitive behavior intervention on the basis of drug treatment. All the students in the two groups completed the self-rated health measurement scale before intervention (T0),after intervention (T1) and 8 weeks after intervention (T2). Results (1) There were no significant differences in total health score and dimension score between the two groups before intervention (both P<0. 05). (2)The repeated measurement variance analysis showed that there was a signif-icant group × time interaction effect on total health score and dimensions(P>0. 05). (3) The group effect of physical health was not significant (P>0. 05). The group effect of total mental health, social health and health score at T1 and T2 time points were significant (all P>0. 05). (4)Compared with before intervention, mental health ((123. 34±9. 33),( 122. 63± 9. 11)),social health ((102. 89 ± 7. 28),( 101. 89± 7. 73)) and total health score ((370. 34±17. 99),(367. 63±17. 89)) of intervention group at T1 and T2 increased ( all P<0. 05),while that of control group increased only at T1 (all P<0. 05). Conclusion Group cognitive behavioral intervention has no obvious effect on physical health of middle school students with emotional dis-orders. And group cognitive behavioral intervention can effectively improve their mental health,social health and overall health level,and the long-term effect is better.
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Objective To explore the effect of group cognitive-behavior therapy intervention on dys-thymic disorders in children and adolescents.Methods From July 2015 to December 2017,68 cases of chil-dren and adolescents treated in Lishui Second People's Hospital were divided into two groups randomly. The study group ( 34 cases) were treated with group cognitive-behavior therapy,and the control group ( 34 cases) were treated with selective 5-hydroxytryptamine reuptake blocker ( SSRI ) . The achenbach child behavior check list(CBCL),self-rating depression scale(SDS) and self-rating anxiety scale(SAS) scores of the two groups before and after the intervention were compared.Results Before the intervention,there was no differ-ence in the score of CBCL,SDS and SAS between the study group and the control group (P>0.05). After drug treatment or cognitive-behavior therapy, the social, activity, thinking and aggressive behavior of both groups were obviously improved. The SDS and SAS scores were 43.64±4.86 and 42.27±3.74 in cognitive-be-havior therapy group,which had significant difference (P<0.05) compared with pre-intervention score of SDS (82.91±3.95) and SAS (78.61±6.28). The scores of SDS (45.61±8.03) and SAS (44.09±7.04) in treat-ment group were significantly (P<0.05) decreased compared with those before treatment (84.03±5.11 and 77.30±9.55,respectively). The satisfaction of the study group and control group was 94.1％ and 67.6％,re-spectively,and the difference was significant between the two groups(P<0.05).Conclusion The effect of cognitive-behavior therapy in children and adolescents on reducing the negative emotion is remarkable,which is equivalent to the drug treatment.The cognitive-behavior therapy in children and adolescents can be used as a auxiliary means to treat children's dysthymic disorders in clinical practice.
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Objective To generate recombinant Pichia pastoris for high-copy expression of human tissue factor pathway inhibitor (TFPI). Methods The cDNA encoding human TFPI was inserted into the expression vector pPIC9K and the constructed expression vector rhTFPI-pPIC9K was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant plasmids were subsequently transformed into Pichia pastoris GS115 cells, and the transformants were confirmed by PCR amplification of the genomic DNA.The recombinant Pichia pastoris with high copies of TFPI cDNA was screened by G418 selection. Western blot and TFPI ELISA Kit were employed to analyzing. The temperature, time and concentration of methanol for the induction of recombinant protein were optimized. Results PCR analysis and DNA sequencing confirmed the successful construction of the expression vector rhTFPI-pPIC9K. Real time quantitative PCR and Western blot analysis demonstrated the positive correlation between TFPI expression level and the copy number of TFPI cDNA in Pichia pastoris cells. Optimization of the induction condition significantly elevated the expression level and activity of TFPI (9.95±0.78 mg/L and 3.91±1.37 U/mL). Conclusion The Pichia pastoris strain with high copy of TFPI expression was successfully constructed, which lays a solid foundation for the further investigation on the function of TFPI and its application in the prevention and therapy of diseases.
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Objective To investigate the expression of MUC5AC in human sinus mucosa and compare the expression of MUC5AC mRNAs in normal and chronic sinus mucosa. Methods Thirty two chronic ethmoid sinusitis mucosa samples and 7 normal ethmoid sinus mucosa samples were used. The protein expression of MUC5AC was examined by using immunohistochemical method. RNAs were extracted from sinus mucosa, and fluorescent quantitative nested RT-PCR was performed for MUC5AC. Results The mean gray scale of MUC5AC protein expression was 159. 72 ± 14. 14 in chronic ethmoid sinusitis mucosa and 115.80 ±31.58 in normal ethmoid sinus mucosa. There was significantly different between two groups (t =3.57, P ＜0.01). The levels of MUC5AC mRNAs in chronic rhinosinusitis [(35.80 ± 19. 74) × 105copies/μg] were higher than those in normal sinus mucosa [(4. 66 ± 2. 47) × 105 copies/μg] . M UC5 AC mRN A expression had significant difference between chronic ethmoid sinusitis mucosa and normal sinus mucosa(t =4. 12, P ＜0.01). Conlusion This result suggested that up-regulation of MUCSAC in chronic rhinosinusitis might play an important role in the pathogenesis of sinus hyperesecretion in chronic rhinosinusitis.
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OBJECTIVE@#To investigate the expression of MUC2 in human sinus mucosa and compare the expression of MUC2 mRNAs in normal and in chronic sinus mucosa.@*METHOD@#Thirty-two chronic ethmoid sinusitis mucosa samples and 8 normal ethmoid sinus mucosa samples were obtained; RNAs were extracted from sinus mucosa, and fluorescent quantitative PCR was performed for MUC2.@*RESULT@#The levels of MUC2 mRNAs in chronic rhinosinusitis were significantly increased compared with those in normal sinus mucosa.@*CONCLUSION@#This result suggest that up-regulation of MUC2 in chronic rhinosinusitis may play an important role in the pathogenesis of sinus hypersecretion in chronic rhinosinusitis.