ABSTRACT
Objective To investigate the changes of cannabinoid 2 receptor(CB2R) expression at different time after epileptic status in epileptic childhood rats. Methods Healthy male SD rats (18-21 days old) were randomly divided into two groups:control group and epileptic model group. CB2R concentration in hippocampus of rats were tested at 2 h,24 h,14 d,21 d after entering the status epilepticus( status epilepsy, SE) by immunohistochemistry,PCR,Western blot. Results In control group,CB2R content of the hippo-campus brain gradually increased with age increasing. When the rats with the age of 35-42 days,CB2R con-tent gradually got stabilized. After status epilepticus for 2 h-14 d,CB2R content of hippocampus in epileptic model group was more than that in the control group. At the point of 21 d,CB2R content of hippocampus in the control group was more than that in epileptic model group. CB2R mRNA of epileptic model group at 2 hours point was more than that of control group (2. 062 vs 1. 878,P<0. 05). At 24 h and 14 d after SE,there were significant differences between two groups in CB2R mRNA respectively ( P <0. 05, respectively). At 21 d after SE,CB2R mRNA of control group was more than that of epileptic group (6. 018 vs 5. 938),but there was no significant difference between two groups (P >0.05). Conclusion CB2R presents high expression in childhood rat hippocampus suffering from status epilepticus, reaches a peak following with prolonged seizures,then gradually decreased.
ABSTRACT
Thyroid carcinoma is the most common malignant tumor in the endocrine diseases,with a rising morbidity.As more investigations were made,thyroid stimulating hormone receptor is showed up,and it is believed some contacts are existed between thyroid stimulating hormone receptor and thyroid carcinoma.We believe that making sure of these contacts can help patients in diagnosis,treatment,and prognosis.
ABSTRACT
Objective:To investigate mitochondrial-dependent molecular mechanisms of a novel agonistic anti-human death receptor 5 (DR5) monoclonal antibody(mDRA-6) inducing apoptosis in Jurkat cell.Methods:The dose-dependent and time-dependent cell growth suppression of mDRA-6 in Jurkat cells was determined by MTT assay.The measurement of the mitochondrial transmembrane potential(ΔΨm) of Jurkat cells was detected by flow cytometry with JC-1 single staining.Caspase-8,9 as well as Bid,Bax,Bcl-2 and Cyto c of apoptotic Jurkat cells were analyzed by Western blot after mDRA-6 treatment.Results:The mDRA-6 induced cell growth suppression and cytotoxicity in dose-dependent manner and time-dependent manner.After mDRA-6 treatment at 2.0 μg/ml for15 min,30 min,60 min and 120 min,the change in ΔΨm were 20.14 %,19.34 %,21.11% and 30.90% respectively by JC-1 single staining.Western blot revealed that the level of active fragments of Caspase-8,9 and Bid,Bax,Bcl-2 and Cyto c respectively,and the amount of Cyto c was increased in cytosol concomitant with the related attenuation of Cyto c in mitochondria.Conclusion: Apoptotic pathway of Jurkat cells induced by mDRA-6 is initiated upon DR5 ligatian to mDRA-6 and exogenic Caspase-dependent cell apoptotic cascades is activated,and endogenic mitochondrial-dependent cell apoptosis pathway is activated.mDRA-6 may be a useful agent in investigating human leukemia therapy by using TRAIL/DR5.
ABSTRACT
Objective To investigate the expression and significance of IL-6 and IL-6 receptor (IL-6R)in hysteromyoma.Methods The expression of IL-6 in hysteromyoma and homologous normal myometrium of 20 cases undergoing total hysterectomy were detected by fluorescent quantitative PCR and western-blot,respectively while IL6R was examined by western-blot and RT-PCR.Results The expression of IL-6 protein and mRNA in homologous normal myometrium were obviously higher than that in hysteromyoma[(0.7996 ±0.0359)vs (0.6904 ±0.0414)and (0.0470±0.0071)vs(0.0283±0.0045 ),P＜0.05] ;No difference was observed in IL-6R protein and mRNA expression between leiomyomas and myometrium[(0.7105 ±0.0520)vs (0.6904 ±0.0532)and (0.6644 ±0.0537 )vs (0.6465±0.0501),P＞0.05 ].Conclusion The lower expression of IL-6 in leiomyomas may contribute to the pathogenesis of hysteromyoma to some extent,and there is no relationship between IL-6R and hysteromyoma.
ABSTRACT
Objection:To construct and characterize TNFR1-GFP fusion protein expression plasmid(pEGFP-TNFR1) and provide a direct and simplified method for assessment of the effect of TNFR1(siRNA) on the TNFR1 gene expression. Methods:Total RNA was extracted from mouse liver.TNFR1 cDNA was amplified by RT-PCR technique and cloned into pMD18-T vector and the sequence was ensured by sequenciog assay.TNFR1 gene was released from pMD-TNFR1 and subcloned into pEGFP-N2 upstream of GFP gene.pEGFP-TNFR1 was analyzed by restrictive enzymatic digestion to ensure the orientation.The plasmid was then transfected into CHO cells,then the fusion protein expression was detected by immunohistochemistry and fluorescent microscope.Rusults:A 1360 bp gene fragment was obtained and coloned into pMD18-T vector,and the sequence was correct.TNFR1 gene was subcloned into pEGFP-N2 vector,and then restriction endonucleases assays showed the correct orientation.24-48 hours after transfection in CHO cells,the expression of TNFR1-GFP fusion protein can be detected by immunohistochemistry and fluorescent microscope.Conclusion:pEGFP-TNFR1 has been constructed successfully.The TNFR1-GFP fusion proteinwas expressed in CHO cells after transtection.It may provide a direct and simplified method for primary assessment of the effect of TNFR1 siRNA on the(TNFR1) gene expression.
ABSTRACT
Objective To investigate chemokine receptor CX3CR1 gene T280M polymorphism in patients with ischemic cerebrovascular disease(ICVD) and its frequency.Methods 165 patients with ICVD(cerebral infarction 85 cases,lacunar infarction 40 cases,transient ischemic attack 40 cases) and 150 age- and sex-matched healthy controls(normal control group) were involved in this study.The polymorphism of T280M was analyzed by multiplex polymerase chain reaction-restriction fragment length based(PCR-RFLB),the gene frequency was compared between two groups and each ICVD subgroups.Results There were TT and TM genotypes in normal control group,and TT,TM and MM genotypes in ICVD group.The genotype were significant differences between the two groups(P0.05).Conclusions There is MM genotype in the chemokine receptor CX3CR1 gene T280M in the patients with ICVD,and their M allele frequency obviously increase.It suggests that CX3CR1 gene T280M polymorphism may be associated with ICVD.
ABSTRACT
Objective: To study tumor chemotherapy on IL-2R express in peripheral blood cells of patients with malignant tumors.Methods: CD + 25 cells percentage rate of peripheral blood cells in patients with or without chemotherapy was done.Results: CD + 25 cells percentage rate in malignant tumor was lower than those of normal contrlos (P
ABSTRACT
Hydrogen peroxide (H2O2)-induced aggregation of calf platelets and its modification by agents with specific properties were characterized employing a spectrophotometric assay. An Arrhenius activation energy of 20 ± 1 kcal/mol was found in the temperature range of 25°-36°C. Rate inhibition occurred on either side of this temperature range, and under anaerobic conditions. Exogenous Ca2+ ions were not required but Ca2+ ions, at 1 mM-concentration, optimally increased rates and extent of aggregation at suboptimal H2O2 concentrations but only extent of aggregation at optimal H2O2 concentrations. Ba2+, Sr2+, Cd2+, Mn2+ and Ni2+ ions (1 mM) and Zn2+, Pb2+ and Hg2+ ions (10 mM) were inhibitory. The cyclo-oxygenase inhibitor, indomethacin (10- 30 mM) exerted only mild inhibition by a competitive mechanism. Another cyclooxygenase inhibitor, aspirin, functioned to increase aggregation. Ligands acting directly at the prostaglandin H2/thromboxane A, receptor (5Z. 9, 11, 13E, 15(S) 15-hydroxy 9(11) epoxy methano prosta 5, 13-dien-1-oic acid, pinane thromboxane A2, arachidonic acid, eicosapentaenoic acid, and N-ethylmaleimide) functioned as competitive inhibitors. Another platelet-activating sulphydryl reagent, thimerosal, also inhibited competitively while the protein kinase C inhibitor, sphingosine, and the protein kinase C modulator, Zn2+ ions, inhibited by different mechanisms. The results indicate direct action of H2O2 at the prostaglandin H2/thromboxane A2 receptor, possibly its sulphydryls, to activate the protein kinase C pathway, independently of cyclo-oxygenase products. The results underscored the power of the kinetic approach for investigating mechanisms of platelet activation.
ABSTRACT
The present study was to investigate the analgesic effect and mechanism of meto-clopramide (MCP) on a rabbit visceral pain model. The results showed that MCP (8 mg? kg-1,iv) could produce a significant analgesic effect on visceral pain (P