Objective To develop an ELISA for detection of anti-dsDNA antibody by using plasmid DNA as antigen.Methods DNA in plasmid pBV220 for prokaryotic expression vector was purified by base-cleavage. The microtiterplates were pretreated by poly-L-lysine and coated by the plasmid DNA in a dilution of 1∶50 as antigen. An ELISA method for detection of serum anti-dsDNA antibodies was developed with HRP-SPA as enzyme-labeled marker.The IIF using crithidia lucilia as substrate was performed simultaneously for comparison. The serum samples from 64 patients with SLE, 8 with MCTD and 17 with RA were detected. Results The concentration of DNA was 1 54 g/L by UV spectrophotometer at wavelength of 260 nm. The positive percentage of ELISA for anti-dsDNA was higher than that of IIF. By comparison with IIF the positive percentages in SLE, MCTD and RA groups were 23 4% vs 17 2%, 12 5% vs 12 5% and 11 8% vs 5 9%, respectively, and the coincident rates between the 2 methods were 93 8%, 100% and 94 1% respectively. The sensitivity and specificity of the developed ELISA for detection of anti-dsDNA were 100% and 93 4% when IIF was as gold standard.Conclusion The ELISA by using plasmid DNA as antigen to detect anti-dsDNA has fine precision, sensitivity and specificity. Its positive rate is higher than that of IIF thus it will contribute to monitor the activities for SLE patients′ condition.

Objective To investigate the expression of CD40 and CD40L on the surface of peripheral blood mononuclear cells(PBMCs)in asthmatic rats and the effect of anti-CD40L McAb on cytokines of it.Methods Flow cytometry and RT-PCR were used to detect the expression of CD40 and CD40L of PBMCs ih asthmatic rats.After the PBMCs Was treated with anti.CIMOL McAb.ELISA was used to detect the levels of IL-4 and IFN-γin the supematants of cultured cells.Results Compared with the normal control group.the expression of CD40 and CD40L of PBMCs in asthImatic rats increased(P＜0.05).Compared with the untreated group,the level of IL-4 and the ratio of IL4/IFN-γ decreased after the PBMCs were treated with anti-CD40L McAb(P＜0.05).Conclusion The expression of CD40 and CD40L on the surface of PBMCs in asthmatic rats Was unregulated.Anti-CD40L Mcab Can decrease the level of IL-4 and the ratio of IL_4/IFN-γ.

Adenocarcinoma, Clear Cell ; metabolism ; pathology ; surgery ; Aged ; Diagnosis, Differential ; Female ; Humans ; Keratins ; metabolism ; Male ; Mucin-1 ; metabolism ; Palatal Neoplasms ; metabolism ; pathology ; surgery ; Palate, Hard ; surgery ; Tongue Neoplasms ; metabolism ; pathology ; surgery

Adrenal Glands ; Female ; Hematopoiesis, Extramedullary ; Humans ; Spherocytosis, Hereditary ; complications ; Young Adult

Glioma ; pathology ; Humans ; Neoplastic Stem Cells ; pathology

Objective To quantitatively analyze the coronary artery ostia by three-dimensional trans-esophageal echocardiography (3D-TEE).Methods The full-volume images of aortic root and coronary artery ostia were acquired by 3D-TEE in 95 adult patients.The Philips QLab 3DQ measurement technology was employed to determine three mutually perpendicular planes:(1) The transverse plane cross the bottom of three coronary artery sinus.(2) The sagittal plane perpendicular to sino-tubular junction.(3) The coronal plane perpendicular to the aforementioned two planes .The following relevant parameters were measured and recorded:(1) Length, width, height and area of bilateral coronary artery ostia .(2) The angle between coronary arterial outflow tract and aortic root in sagittal plane .(3) The spatial distribution of coronary artery ostia, aortic root and coronary artert sinus .Results The shape of left coronary artery ostia were more regular (round or oval) than right coronary artery ostia ( teardrop-shape or oval ).Calcification was more frequent in right coronary artery ostia (81/95, 85.26%) than that in left coronary artery ostia. There were statistical differences between left and right coronary artery in the parameters of ostial wide , area and height (t =3.85, 3.86, -4.49, all P<0.01).Most left coronary artery ostia were located inside the sinus (76/95, 80.00%), mainly in the upper third segment (69/95, 72.63%); while more than half of the right coronary artery ostia were found outside the sinus ( 53/95, 55.79%).The difference was statistically significant( χ2 =25.91, P<0.01).Conclusion The quantitative analysis of aortic root and coronary artery ostia based on the full-volume images originated from real-time 3D-TEE is feasible, which is helpful for further clinical research .

Objective To explore the association between L-selectin gene P213S polymorphism and angina pectoris.Methods L-selectin gene P213S polymorphism in 138 patients with angina pectoris and 156 controls was detected by polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP).The relationship between gene polymorphism of L-selectin and levels of serum lipids were also studied.Results L-selectin genotype frequencies of PP,PS,SS were 60.1%,36.3%,3.6% and 44.9%,48.1%,7.0% in angina pectoris group and control group respectively.Allele frequencies of P,S were 78.3%,21.7% and 68.9%,31.1% in angina pectoris group and control group respectively.There was significant differences of frequencies of genotype and allele of L-selectin P213S polymorphism between angina pectoris group and control group(P

Objective: To investigate the effect of Atractyloside on sevoflurane postconditioning in focal cerebral ischemia.Methods: Fifty male SD rats were randomly assigned to five groups(n=10 each): control group(con),1.0 MAC group(sevo),1.0 MAC+ Atractyloside group(sevo+Atr),1.0 MAC+ vehicle group(sevo+vehicle)and Atractyloside group(Atr).All animals were subjected to the right middle cerebral artery occlusion(MCAO) for 120 min followed by reperfusion for 72 h.The animals in sevo groups were given 1.0 MAC sevoflurane inhalation from 20 min before to 10 min after reperfusion.The animals in Atr groups were given Atractyloside by ICV injection before sevoflurane postcondioning.The neurological deficit scores(NDS) were recorded at 24 h,48 h,and 72 h after reperfusion.Infarct volume percentage was determined after the last NDS assessment.Results: The infarct volume percentage ratio of sevo group(0.32?0.05) was significantly lower than that of con group(0.55?0.07)(P0.05).The infarct volume percentage ratio of sevo+Atr group and Atr group were significantly higher than that of Sevo group(P0.05).NDS of 24 h,48 h and 72 h after reperfusion in sevo group and sevo+vehicle were significantly higher than that in Con group.NDS of 24 h,48 h and 72 h after reperfusion in sevo+Atr group and Atr were significantly lower than that in Sevo group.Conclusion: Atractyloside abolished the protective effects of sevoflurane postconditioning on focal cerebral ischemia-reperfusion injury.

Objective:Ischemic cerebral injury is a common complication during perioperative period.We found that ischemic postconditioning and various non-ischemic postconditioning(e.g.inhalation anesthetics) could significantly attenuate ischemic cerebral injury.The present study was to investigate the effect of Cyclosporin A on sevoflurane postconditioning in focal cerebral ischemia.Methods:Fifty male SD rats were randomly assigned to five groups (n=10 each): control group(con),1.0MAC group(sevo),1.0MAC+ Cyclosporin A group(sevo+CsA),1.0MAC+ vehicle group(sevo+vehicle)and Cyclosporin A group(CsA).All animals were subjected to right middle cerebral artery occlusion(MCAO) for 120min followed by reperfusion for 72h.The animals in sevo groups were given 1.0MAC sevoflurane inhalation from 20min before to 10min after reperfusion.The animals in CsA groups were given CsA by ICV injection before sevoflurane postconditioning.The neurological deficit scores(NDS) were recorded at 24h,48h,and 72h after reperfusion.Infarct volume percentage was determined after the last NDS assessment.Results:The infarct volume percentage ratio of Sevo, Sevo+CsA,CsA and Sevo+vehicle groups were 0.31?0.04,0.25?0.04,0.30?0.03 and 0.33?0.05,respectively(P

Objective To establish an LC-MS/MS method for determination of S-071031 B, a novel antidepressant , in rat plasma and to study its pharmacokinetic profiles .Methods An LC-MS/MS method was established to determine S-071031B in rat plasma, and L-8021 was employed as the internal standard .The analytes were separated on a C18 column with a mobile phase consisting of water-acetonitrile containing 0.1%(v/v) formic acid at a flow rate of 0.3 ml/min.The mass spectrometer was operated in a selected reaction monitoring ( SRM ) mode with a positive electrospray ionization (ESI) interface.The plasma concentration-time curve was drawn and pharmacokinetic parameters were calculated by DAS 2.0.Results The linear range was from 2 to 1000 ng/ml with a sensitivity of 2 ng/ml as the lower limit of quantification . The intra-day and inter-day precisions , recoveries and matrix effects at three spiked levels were all suited to the determina-tion of biological samples.After oral administration of S-071031B, the Cmax of S-071031B was (287.2 ±50.8) μg/L and the Tmax was (0.8 ±0.3) h, with a t1/2of (2.9 ±0.6) h and an AUC(0-∞)of (1372.6 ±255.3) μg/L· h.Conclusion This method is sensitive and specific enough for determination of S-071031 B in rat plasma to facilitate the study of its phar-macokinetics .