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Objective:To investigate the relationship between anti-Helicobacter pylori antibody(Hp-IgG)and anti-aquaporin 4 antibody which are in neuromyelitis optica(NMO)and multiple sclerosis(MS).Methods: Serum specimens were collected from the 33 patients with MS,7 patients with NMO,and 35 health examination cases.Hp-IgG were detected by enzyme-linked immunosorbent assasy and anti-aquaporin 4 antibody were detected by cell based assay respectively.The positive rate of Hp-IgG and anti-aquaporin 4 antibody were analyzed,and the difference of Hp-IgG positive rate was compared between patients with Hp-IgG positive and negative.Results: Serum Hp-IgG positive rate of MS,NMO and normal control groups were 69.70％,85.71％ and 42.86％ respectively with a significant statistically difference of Hp-IgG(P0.05).Serum anti AQP4 antibody positive rate of MS,NMO and normal control groups were 4.2％,85.71％ and 0％ respectively with a significant statistically difference of anti AQP4 antibody(P0.05).Conclusion: HP infection is a risk factor for the occurrence of MS and NMO,but not associated with MS and NMO patients with anti AQP4 antibodies.
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Objective:To evaluate immune response of murine peritoneal macrophage challenging by methicillin-resistant S.aureus(MRSA)after pretreatment with Pam3Csk4(TLR2 agonist).Methods: Murine peritoneal macrophage was pretreated with Pam3Csk4(1 μg/ml).Following pretreatment 12 h later,heat-killed MRSA( HK-MRSA) was added and incubated for another 2 or 6 hours.The protein and mRNA level of TNF-α, IL-6 and IL-1 were determined by ELISA and Q-PCR, respectively.To estimate phagocytosis of macrophage,HK-MRSA/MSSA labeled with FITC( FITC-HK-MRSA/MSSA) were added to well and incubated for 30 min.After washing 5 times with PBS,intracellular FITC-HK-MRSA was detected by flow cytometry.To estimate antimicrobal activity of macrophage,live MRSA and MSSA were added to well and incubated at indication time,the CFU of s.aureus was estimated via a 10-fold serial dilution on agar media.cDNA was further quantitative assessed using primers for mouse FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 by Q-PCR .Results: Compared with saline-pretreated cell, the protein and mRNA level of TNF-α, IL-6 and IL-1 were markely reduced, respectively.However, both the phagocytosis and antimicrobal activity to S.aureus were significantly increased in macrophages pretreated with Pam3Csk4.Further study found that the macrophages had higher FCR-Ⅰ,FCR-Ⅲ,CR-1,CR-3,iNOS and LL37 expression at 6 h and 12 h post-stimulation Pam3Csk4.Conclusion: The results suggest that Pam3Csk4 could activate murine antimicrobal activity of peritoneal macrophage challenging by methicillin-resistant Saureus via increasing opsonophagocytosis in depended antibodies, complements manners.The results suggest Pam3Csk4 probably be a novel immunotherapy candidate against MRSA.
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Objective To investigate the imaging features and the treatments of ischemic cerebrovascular disease with unruptured intracranial aneurysms.Methods The clinical data of 23 patients of ischemic cerebrovascular disease with unruptured intracranial aneurysms ( unruptured group) and 147 patients of hemorrhagic cerebrovascular with ruptured intracranial aneurysms ( ruptured group) were retrospectively analyzed.Results There were no statistical significances between the two groups baselines such as age, gender, hypertension, diabetes, hyperlipidemia, heart disease, smoking, drinking alcohol (all P>0.05).There were 16 patients (69.6%) with single intracranial aneurysm and 7 patients (30.4%) with multi-intracranial aneurysms in the unruptured group, and the number of intracranial aneurysms were 34.There were 125 patients (85%) with single intracranial aneurysm and 22 patients ( 15%) with multi-intracranial aneurysms in the ruptured group, and the number of intracranial aneurysms were 170.There was no statistical significance of single aneurysm and multi-aneurysms percentage between the two groups (P>0.05).The maximum aneurysm diameter and the percentage of large aneurysms, dissecting aneurysms of the unruptured group were significantly greater than the ruptured group; and the percentage of small-middle aneurysms and saccular aneurysms were significantly smaller than the ruptured group ( all P<0.05 ) .In the unruptured group, 5 cases were treated with intervention treatment and 1 case was treated with aneurysm clip surgery, all of which recovered well after surgery;6 cases were treated with Plavix antiplatelet therapy, 5 cases were treated with Aspirin antiplatelet therapy, cerebrovascular disease events were not found of which followed-up for 6 months;6 cases were not taken antiplatelet therapy, and 1 case of which followed-up and occurred cerebral infarction.In the ruptured group, 46 cases were treated with intervention treatment, 2 cases of which were died;101 cases were treated with aneurysm clip surgery, 6 cases of which were died.Conclusions Small-middle aneurysms are more commonly occurred in patients of ischemic cerebrovascular disease with unruptured intracranial aneurysm, however, the maximum aneurysm diameter of which is significantly bigger than patiehts of hemorrhagic cerebrovascular with ruptured intracranial aneurysms.Most of unruptured aneurysms which had duty relationship with cerebral ischemic events are dissecting aneurysms and saccular aneurysms.Intervention treatment of unruptured intracranial aneurysms is relatively safe and effective.
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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 .
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Objective To observe therapeutic benefits of intravenously transplanted hone marrow mesenchymal stem cells (BMMSCs) on alcohol-associated dementia (AAD) rat model and study its underlying mechanisms.Methods BMMSCs were isolated by the method of differential adhesion and membrane antigens were detected with flowcytometric analysis.To establish AAD model,SD rats were intragastricly administrated with ethanol (20％,8ml/kg) for 28 days.Then BMMSCs were labeled with DAPI and injected into the blood via caudal vein.And animals were evaluated by observing Morris Maze behavior,hippocampal morphology and neuronal apoptosis.The expression of BDNF was detected by the method of immunohistochemistry.And the activities of total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) in rat blood serum were also measured.Results The results of flowcytometry analysis indicated BMMSCs were CD29,CD90-positive,and CD45,CD34-negative.And the cells labeled with DAPI were observed in rat hippocampus 3 days after intravenous injection.Compared with PBS group,the escape latency of rats in BMMSC group was apparently shortened((10.17 ±0.71)s vs.(4.71 ± 0.34)s,P ＜0.01).And the morphological structure was repaired and neuronal apoptosis was reduced in rat hippocampus after BMMSC transplanting((72.67 ± 2.73) vs.(55.5 ± 5.14),P＜0.05).Immunohistochemical results showed that the expression of BDNF was significantly increased in the hippocampus of rats in BMMSC group ((71.54 ± 13.71) vs.(135.25 ± 22.20),P ＜0.05).Also,the activity of GSH-Px was apparently improved in the blood serum of rats treated with BMMSC transplanting ((526.89 ± 62.73) vs.(2592.75 ±243.73),P ＜0.01),but no change for that of T-SOD.Conclusion The results provide a novel therapeutic strategy for improving learning and memory function and reducing hippocampal damage induced by ethanol administration,which is closely related to enhance BDNF expression in the hippocampus and improve the activity of antioxidants.
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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.
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Objective To investigate the therapeutic effect and the detailed mechanisms of intraarterially delivery of bone marrow mesenchymal stem cells ( BMSCs) for treatment of middle cerebral artery occlusion (MCAO) in rats.Methods BMSCs were isolated,purified and amplified with the adherence culture method.BMSCs were labeled with 5-bromo-2-deoxyuridine ( BrdU ) (10 μmol/L) for 48 h before transplation.Surface antigens of CD90,CD29,CD106,CD34,CD45,CD11b were identified by flow cytometry.The MCAO model was established with suture emboli method.In this study,3×106 BMSCs were injected into rats with MCAO through intraarterial route at day 7 after stroke.The effects on functional and physical recovery were assessed with the behavioral tests (mNSS test and adhesive test) and body weight.Bielshowsky-Luxol Fast Blue double staining was used to demonstrate the reconstruction of axon and myelin.The Brdu-labeled BMSCs in vitro and in vivo were detected with direct immunofluorescent staining.The expression of neuron specific enolase ( NSE),neurite outgrowth inhibitor-A ( Nogo-A),synaptophysin (SYN),ki-67 nuclear antigen (Ki-67),glial fibrillary acid protein( GFAP),vascular endothelial growth factor ( VEGF) in brain were analyzed with immunohistochemical staining.Results Flow cytometry indicated that the positive rates of high expression of CD90,CD29,CD106 in BMSCs were respectively 91.70%,88.40% and 52.20%.Meanwhile,the positive rates of low expression of CD34,CD45,CD11b in BMSCs were 2.70%,5.65% and 7.82%,respectively.There was a significant difference in behavioral tests ( mNSS test and adhesive test) between BMSCs group and PBS group at day 21,28,35 after MCAO (mNSS:4.89 ±1.36,7.00 ±1.67,3.78 ±1.30 and 6.33 ±1.21,2.44 ±1.13,5.67 ± 1.51;t =2.69,3.83,4.75;adhesive test:54.00 ± 10.48,68.17 ± 11.09,36.89 ±9.80 and 59.33 ± 12.40,23.44 ± 9.04,46.50 ±9.38;t =2.51,3.92,4.77;P ＜0.05).Meanwhile,a significant difference in body weight was discovered between them at day 28,35 after MCAO.In BMSCs group,the area of corpus callosum in the ipsilateral hemisphere was significantly enlarged,the positive number of Brdu,SYN,Ki-67,GFAP,VEGF in brain was significantly increased,the expression of Nogo-A in brain was significantly decreased,nevertheless,the number of NSE-positive cells in brain and the infarct volume were not significant different from PBS group at day 35 after MCAO.Conclusions These results suggest that intra-arterial transplantation of BMSCs is an efficient treatment protocol for stroke.Treatment with BMSCs increases endogenous cells proliferation,angiogenesis,synaptogenesis,enhances axonal regeneration and the protective function of astrocytes,all of which may contribute to neurological functional recovery.
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Objective To evaluate the detection of culture filtrate protein 10 (CFP10) and 6000 early secretory antigenic target (ESAT-6) in cerebrospinal fluid to be used in diagnosing tuberculous meningitis. Methods Dot enzyme linked immunosorbent assay ( Dot ELISA) method that was improved by applying concentrated cerebrospinal fluid was used to detect CFP10 and ESAT-6 in cerebrospinal fluid to analyze small protein antigen secreted by M. tuberculosis. Cerebrospinal fluid of 111 subjects were collected,in which 58 specimens were clinically diagnosed as tuberculous meningitis and 53 as non-tuberculous.CFP10 and ESAT-6 were detected in cerebrospinal fluid using Dot ELISA method and the results were analyzed. Results The sensitivities of detecting CFP10 and ESAT-6 antigen were 93.1% and 91.4% respectively, and the specificities were 92. 5% and 94. 3% respectively. The sensitivities and specificities are generally higher compared with the other methods of detecting M. tuberculosis or materials of M. tuberculosis by acid-fast staining or mycobacterium tuberculosis culture and polymerase chain reaction.Conclusions Using Dot ELISA method to detect CFP10 and ESAT-6 in cerebrospinal fluid to diagnose tuberculous meningitis has a high sensitivity and specificity. Our study provided the evidence of detecting the specific antigen of M. tuberculosis to be used in diagnosing tuberculosis.
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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.
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Objective To study the relationship between acute renal failure(ARF) and hyperosmolaremia in stroke patients and the related factors.Methods The clinical data of 59 stroke patients with ARF(ARF group) and 76 stroke patients without ARF(control group) were studied.The correlation analysis was used to compare the serum osmolality levels and serum creatinine values between the two groups.The factors that caused ARF in stroke patients were estimated using a logistic regression model.Results The mean serum osmolality in ARF group[((320.98?)30.63)mOsm/L] was significantly higher than that in control group [((295.49?)17.7)mOsm/L]((P
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Objective To explore the differentiation of human bone marrow measenchymal stem cells (hMSCs) into neuron-like and dopaminergic neuron-like cells in vitro.Methods The hMSCs were isolated from adult human bone marrow and expanded on the flask undifferentiated state for 2 passages. After pretreatment with WHI-P131 for 48 h, the hMSCs were cultured at the medium containing 10 ng/ml basic fibroblast growth factor for 24 h, then incubated with all-trans-retinoic acid and glial-derived neurotrophic factor in serum-free media for 5 h. The surface markers of differentiated neuron were detected by immunocytochemical method and the transdifferentiation process was observed under light microscope.Results Under induction conditions, hMSCs progressively resumed typical neuronal morphological characteristics. After hMSCs were incubated in induction medium for 5 h, the percentage of NSE, nestin, GFAP, TH and DAT positive cells were (77.0?5.7)%, (54.2?3.7)%,(8.8?2.4)%, (36.5?15.8)% and (26.0?14.2)%, respectively. There were no positive expressions in the control group.Conclusion The hMSCs are able to differentiate into neuron-like and dopaminergic neuron-like cells in vitro.