Objective:To investigate the mechanism of immunoglobulin Y antibodies(IgY) against tumour necrosis factor alpha(TNF-α) and Interleukin-1beta(IL-1β) in treating allergic bronchial asthma through nebulization inhalation.Methods:The allergic bronchial asthma model was established with Hartley guinea pig by ovalbumin nebulizating inhalation.The animals were randomly divided into 4 groups: normal control group(group A)，allergic bronchial asthma group(group B),0.1% anti-TNF-α and IL-1β IgY treating group(group C),1.0% anti-TNF-α and IL-1β IgY treating group (group D).The animals were killed after treatment being accomplished for 2 h,4 h,8 h,24 h and the lungs were made pathological,which were then stained by hematoxylin-eosin(H.E.).The bronchoalveolar lavage fluid (BALF) was collected and the deposited cells were stained by Wright's.Results:①The histological appearance of lung: In group B the histological structure of alveolar ducts and alveolar walls was damaged,the alveolar space was full of transudate and lots of alveolar epithelial cells and leucocytes.The pulmonary interstitial edema,inflammatory cells infiltration,distorted or dilated capillaries and reducing capillary numbers of effective blood stream were observed in alveolar walls.In group C and group D the damage degree of alveolar ducts and alveolar wall was slighter than that in group B and there were few inflammatory cells in alveolar space.In bronchial lumen and pulmonary alveoli the sticky mucus plug was obviously less in group C and group D than in group B.Moreover,inflammatory cell infiltration was seldom observed aroud bronchia,and restoration of bronchial tunica mucosa epithelium was obviously observed in group C and group D.②The cytology appearance of BALF: In group C and group D the numbers of eosinophils,neutrophils,lymphocytes were significantly fewer （2 h,4 h,8 h,P＜0.05）,however,the number of macrophage was significantly more（2 h,4 h,8 h,P＜0.05）than in group B.Conclusion:The anti-TNF-α and IL-1β IgY can obviously alleviate pathological extent of inflammatory reaction in allergic bronchial asthma of guinea pigs by nebulization inhalation therapy.The therapeutic effect of anti-TNF-α and IL-1β IgY between 0.1% and 1.0% concentration is not obvious difference for pathology changes.

0.05).Combination of NGF with bFGFs(10 U/ml NGF+10 ?g/L bFGF or 5 U/ml NGF+5 ?g/L bFGF) not only promoted the proliferation of HDPCs(P

Objective To investigate the correlation between the glial fibrillary acidic protein(GFAP),neuron-specific enolase(NSE),synaptoghysin (SYN),neurite outgrowth inhibitor-A(Nogo-A)expression and neurological outcome in tissue surrounding the infarct during the recovery after cerebral ischemia-reperfusion injury in rats.Methods A 2-hour middle cerebral artery occlusion(MCAO)and reperfusion model in rats was induced by the intraluminal suture method.The modified neurological severity score(mNSS)was performed at day 28,35,42,and 49.Immunohistochemistry was used to detect the expressions of GFAP,NSE,SYN,and Noga-A in tissue surrounding the infarct.Results The mNSS score decreased gradually over time after cerdnal ischemia-reperfusion injury in rats.Except day 35(5.11±0.737)vs.day 42 (4.54±0.519),and day 42 vs.day 49(4.29±0.488),there were significant differences at all other time points(all P<0.05).The numbers of GFAP positive cells deergased gradually form day 28 to day 49,in which,the numbers of GFAP positive cells at day 42(51.00±13.59)vs.day 49(44.38±11.94) were significantly less than those at day 28(69.00±15.10)(P<0.05).There were no significant differences in the numbers of NSE positive cells at all time points,but their integrated optical density(IOD)increased gradually.There were significant differences between day 28(6 218.57±1 864.25)and day 42(9 414.00±2 491.12) or day 49(12 522.50±3 106.99),and between day 35(7 343.40±1 533.35)and day 49(all P< 0.05).There were no significant differences at all other time points.The SYN express (IOD)increased gradually.and it was significantly lower at day 49(66 503.00±12 834.61)than that at day 28(43 905.14±13 208.59)(P<0.05).The numbers of Nogo-A positive cells decreased gradually,and they were significantly less at day 49(42.13±14.45) than those at day 28(59.57±15.25)(P<0.05).The GFAP expression was positively correlated with the mNSS scores(r=0.993,P=0.007).The NSE(r=-0.954,P=0.044)and SYN(r=-0.992,P=0.008) expression was negatively correlated with the mNSS scores.Conclusion The neurological outcome was associated with the downregulation of GFAP expression and the upregalation of NSE and SYN expression during the recovery after cerebral ischemia-reperfusion injury in rats.

Objective:To explore the mechanisms of the inhalation of atomized 1.0% anti-IL-1βand TNF-αimmunoglobulin yolk ( IgY) on treating guinea pigs with allergic asthma induced by the inhalation of aerosolized ovalbumin ( OVA).Methods:Healthy guinea pigs were randomly divided into the normal controls ( group C ) , the allergic asthma model group ( group M )-treated by the inhalation of atomized ovalbumin ( OVA ) , the inhalation of atomized 1.0% anti-IL-1βand TNF-αimmunoglobulin yolk ( IgY ) treatment group (group Z1)-treated asthma model guinea pigs by the inhalation of atomized 1.0% anti-IL-1βand TNF-αIgY,and positive control the inhalation of atomized budesonide treatment group (group Z2)-treated asthma model guinea pigs by the inhalation of atomized budesonide.The blood was gotten by cardiac puncture and the bronchoalveolar lavage fluid ( BALF ) was collected by bron-choalveolar lavage at 2 h,4 h,8 h and 24 h after the last time atomization.The inflammatory cells in the peripheral blood ( PB) were counted by methylene blue and eosin staining.Cytokine concentrations of IL-1β,IL-4,IL-6,IL-8,IL-13,IL-16,TNF-α,TGF-β1 and IgE in the plasma and bronchoalveolar lavage fluid (BALF) were measured using an enzyme-linked immunosorbent assay (ELISA).Results:In PB,eosinophils was decreased from 2 h to 8 h in group Z1 compared to group M.In plasma,the levels of IL-1βat 4 h and 24 h,IL-16 at 2 h,4 h and 24 h,TGF-β1 from 4 h to 24 h and IgE at 24 h,as well as the levels of IL-1βand TNF-αfrom 2 h to 8 h,IL-4,IL-6,IL-8 and IL-13 from 4 h to 24 h,IL-16 at 8 h,and TGF-β1 and IgE from 4 h to 8 h,especially the level of IL-1βand TNF-αstarting at 2 h,in BALF were significantly reduced in group Z 1 compared to group M ( P<0.05 ).The levels of IL-1βand TNF-αwere positively cor-related with that of IL-4,IL-6,IL-8,IL-13,IL-16,TGF-β1 and IgE (P<0.05).Conclusion: The inhalation of aerosolized anti-IL-1βand TNF-αIgY effectively alleviates inflammatory responses in guinea pigs with allergic asthma induced by aerosolized OVA inhalation may be due to the significant decrease in the levels of various allergic inflammatory cytokines .

Objective:To investigate therapeutic mechanism of immunoglobulin Yolk (IgY) against tumour necrosis factor alpha ( TNF-α) and interleukin-1 beta ( IL-1β) in guinea pigs with allergic rhinitis.Methods: Hartley guinea pigs were randomly divided into the control group (group C,n=17),the allergic rhinitis model group (group M,n=27),the 0.1%anti-TNF-αand IL-1βIgY treating group (group Z1,n=21) and the fluticasone propionate treating group (group Z2,n=21).The allergic rhinitis model in guinea pigs was established using ovalbumin.After treatment for 2 h,4 h,8 h,nose and bronchial lung were lavaged using 0.9%saline, the nasal lavage fluid (NLF) and bronchoalveolar lavage fluid (BALF) were collected,the precipitated cells were stained using Wright′s,the nasal mucosa and lung tissues were stained using methylene blue and eosin (HE),and TNF-α,IL-1β,IL-5 and IL-33 in nasal mucosa and lung tissues were stained using immunohistochemistry.Results:There were a large amount of eosinophils and more serious inflammation responses in nasal mucosa in the M group compared with the Z 1 and Z2 groups.In the lung tissues,there were more alveolar tube damage ,pulmonary interstitial edema ,interval thickening ,thickening of bronchial smooth muscle and inflammation cell in-filtration in the M group compared with the Z 1 and Z2 groups.The eosinophils ,lymphocytes and neutrophils were significantly decreased in NLF and BALF in the Z1 and Z2 groups compared with the M group (P<0.05).The expressions of IL-1βand TNF-αfrom 2 h to 8 h and IL-5 and IL-33 from 4 h to 8 h significantly decreased in the nasal mucosa and lung tissues in the Z 1 group compared with the M group ( P<0.05 ).Conclusion:The allergic rhinitis in guinea pigs accompany with the allergic asthma.The inhibitory capacity of anti-TNF-αand IL-1βIgY on pathological responses in guinea pigs with allergic rhinitis may be due to the significant decrease in the infiltration of eosinophils and the expressions of inflammatory cytokines in the nasal mucosas and lung tissues .

Objective To study the correlation between the renin-angiotensin-aldosterone system angiotensinogen (AGT) gene M235T,angiotensin Ⅱ type 1 receptor (AGTR1) gene Al166C,aldosterone synthase (CYP11B2) gene -344C/T polymorphisms and large-artery atherosclerotic (LAA) stroke in a southern Chinese Han population.Methods Polymerase chain reaction and gene sequencing technology were used for the genotyping in patients with LAA and normal controls with AGT gene M235T,AGTR1 gene A1166C,and CYP11B2 gene - 344C/T polymorphisms in a southern Chinese Han population,and to determine the correlation between the 3 gene polymorphisms and LAA by binary logistic regression analysis.Results A total of 107 patients with LAA and 142 healthy controls were included in the study.The frequencies of the AGT gene 253TT genotype (66.36％ vs.50.70％,x2 =6.122,P =0.047) and T allele (79.44％ vs.70.07％ ％,x2 =5.581,P =0.018) in the LAA group were significantly higher than those in the control group.The frequencies of the AGTR1 gene 1166CC genotype (0％ vs.0％,x2 =1.494,P =0.222) and C allele (7.48％ vs.4.93％,x2 =1.399,P =0.237) in the LAA group were no significantly differences with those in the control group.The frequencies of the CYP11B2 gene - 344CC genotype (9.35％ vs.4.23％,x2 =3.603,P =0.165) and C allele (27.10％ vs.26.06％,x2 =0.069,P =0.793) in the LAA group were no significant differences with those in the control group.Binary logistic regression analysis showed that there was no significant correlation between the three gene polymorphisms and the simple LAA diseases.The frequencies of AGT gene 235TT genotype (68.00％ vs.41.90％,x2 =12.446,P =0.002) and T allele (79.33％ vs.64.76％,x2 =8.993,P =0.003) in the LAA patients complicated with hypertension were significantly higher than those in the normotensive control group.Logistic regression analysis showed that the odds ratio (OR) exposed to TT genotype was 2.153 (95％ confidence interval [CI] 0.789-5.872).The OR of T allele was 2.089 (95％ CI 1.285-3.396).Conclusions The AGT gene M235T polymorphism is not associated with the simple LAA in the southern Chinese Han population,but it may be associated with the risk of LAA complicated with hypertension;CYP11B2 gene -344C/T polymorphism and AGTR1 gene A1166C polymorphism are not associated with the onset of LAA in the southern Chinese Han population.

Objective:To explore the influence on inflammation cytokines for anti-IL-1βand TNF-αIgY intranasal treatment in guinea pigs with allergic rhinitis.Methods:The allergic rhinitis model in guinea pigs was established using ovalbumin(OVA).Hartley guinea pigs were randomly divided into the control group(group C,n=17),the allergic rhinitis model group(group M,n=27),the 0.1%anti-IL-1βand TNF-αIgY treatment group(group Z1,n=21)and the fluticasone propionate treatment group(group Z2,n=21). At 2 h,4 h and 8 h after the last treatment,blood was got by heart puncture,as well as nose was lavaged using 0.9% saline and the nasal lavage fluid( NLF) was collected.The level of cytokines was examined using ELISA kits.Results: In the peripheral blood, the levels of IL-1β,IL-5,IL-9,IL-13,IL-18,IL-33 and TGF-β1 from 2 h to 8 h;TNF-αand OVA-specific IgE from 2 h to 4 h;and IL-22 from 4 h to 8 h were significantly decreased in the 0.1%anti-IL-1βand TNF-αIgY treatment group compared with the allergic rhinitis model group(P<0.05).In the NLF,the levels of IL-1β,IL-5,IL-9,IL-13,IL-22,IL-33,TNF-α,TGF-β1 and OVA-specific IgE from 2 h to 8 h;and IL-18 at 2 h were significantly decreased in the 0.1% anti-IL-1βand TNF-αIgY treatment group compared with the allergic rhinitis model group ( P<0.05 ) .Conclusion: Anti-IL-1βand TNF-αIgY intranasal treatment can significantly reduce inflammation cytokine levels in allergic rhinitis guinea pigs.

ObjectiveTo study the effect of extract of fetal brain (Nerval Active Factors, NAF) on learning, memory and stamina of mice.Methods51 mice were divided into five groups randomly: normal saline as Group A, Cerebrolysin as Group B, NAF as Group C:0.1mg,Group D:0.2mg,Group E:0.4mg, i.p.The time looking for food and error frequency of walking road were tested by using complicated maze, and the drowning time was tested when the mice carried 5% body weight on its back.ResultsBefore and after treatment,the time looking for food in maze showed no significant change in Group A(P>0.1) and was significantly shorter in Group B, C, D and E(P<0.005,P<0.05,P<0.001,P<0.001).There was no significant difference on error frequency of walking road in Group A and B(P>0.2,P>0.5) before and after treatment, while it was significantly decreased in mice of Group C, D and E(P<0.02,P<0.02,P<0.01).There was significant difference on drowned time of mice carrying 5% body weight on back in five groups by analysis of variance (P<0.05) after treatment, but this difference derived from C vs E(P<0.05)and D vs E(P<0.01).Conclusions NAF can raise the activity of learning and memory of mice, as well as increase their stamina. On the other hand, the stamina of mice decreases when NAF is given more than 20 mg/kg body weight.

Objective To identify the role of phosphatidylinositol-3-kinase(PI3K) in mediating necroptosis induced by tumor necrosis factor alpha (TNFα) and the involved mechanism.Methods Knockdown of p110α,receptor-interacting protein 1(RIP1) or both p110αand RIP1 was mediated by the specific short hairpin RNA (shRNA) lentivirus and verified by RT-PCR or Western blotting .In addition , Western blotting was used to detect phosphorylation of mixed lineage kinase domain-like protein(MLKL) and protein kinase B(AKT) or tetramerization of MLKL.Cell death was measured by micros-copy and flow cytometry.Results AKT phosphorylation and TNFα-induced necroptosis of L929 cells were suppressed by the inhibitors of PI3K or AKT, as well as p110αknockdown.Moreover, RIP1 knockdown did not inhibit L929 cell death induced by TNFαplus Z-VAD, but the RIP1-independent necroptosis was inhibited by p 110αknockdown.In addition, p110αknockdown suppressed MLKL phosphorylation and tetramerization induced by TNFαwith Z-VAD in L929 cells. Conclusion PI3K mediates necroptosis of L929 cells induced by TNFαby activating AKT and MLKL, respectively.

Objective To investigate the effect of heat stress on the expression of Toll-like receptor 4 (TLR4) and peripheral blood T regulatory cells (Treg) in splenic cells of rats.Methods Senventy-two SD rats were divided into 20 ℃ control group and 37 ℃ group.Each group was divided into non stimulation,bacterial lipopolysaccharide (LPS) stimulation and concanavin A (Con-A) stimulation subgroup.Each subgroup had 1,12,48 h and 168 h observation points,and flow cytometry was used to determine the level of TLR4 and Treg.Results The TLR4+ immunocompetent cells in the spleen was decreased from 1 h to 168 h in every subgroup of the 37 ℃ group compared to the 20 ℃ control group (P＜0.05).The level of CD4+CD25+ Treg in the peripheral blood in rats from 1 to 48 h was significantly decreased (P＜0.05) and slightly increased at 168 h in LPS stimulation subgroup in the 37 ℃ group compared to the 20 ℃ control group.The level of CD4+ CD25+ Foxp3+ Treg in the peripheral blood in rats at 12 h were significantly increased in non-stimulation and concanavalin A (Con-A) stimulation subgroups in the 37 ℃group,and were significantly decreased at 168 h in every subgroup of the 37 ℃ group compared with 20 ℃ control group (P＜0.05).The level of CD8+CD25+ Treg in the peripheral blood in rats was significantly increased at 1 h and 168 h in the 37 ℃ hot and humid group in the every subgroup whencompared with the 20 ℃ control group (P＜0.05).The level of CD8+ CD25+Foxp3+ Treg in the peripheral blood in rats was significantly decreased at 1 h and 48 h in the every subgroup,and was significantly increased at 12 h and 168 h in the non-stimulation and LPS stimulation subgroups in the 37 ℃ group compared to the 20 ℃ control group (P＜0.05).Conclusion High temperature and damp heat can destroy the innate immunity and alter the functional state of adaptive immunity in rats.