Objective To investigate the effect of tripterygium glycosides on the resistance to immune rejection after allogeneic islet transplantation in mice.Methods Twenty C57BL/6 mice were treated with STZ diabetes mellitus and transplanted the islets from Balb/c mice donor,then recipient mice were randomly divided into two groups:triptolide group and model control group(n=10),and were intraperitoneal injected with tripterygium glycoside solutin and equivalent solvent of 5 mg·kg-1·d-1 for 14 days.Blood glucose and body weight were measured within 4 weeks after transplantation.Two weeks later,two groups of grafted islets were stained by HE staining and immunohistochemical staining,the expression of IL-2 protein were detected by Western blotting.Results The level of blood glucose were decreased to normal in the triptolide group and model control group after islet transplantation,but blood glucose gradually increased in the model control group after two weeks.Compared with the model control group,the inflammatory cells were less infiltrated and the immunohistochemical staining of insulin was deeper in the triptolide group.The expression of IL-2 in the triptolide group was significantly decreased(P<0.05).Conclusion Tripterygium glucoside could significantly decrease the inflammatory cell infiltration and inflammation factor expression in the allogeneic islet recipients to reduce the immune rejection and improve graft survival.

Objective To design an experiment for method-comparison and bias estimation of multi tests on multi instruments.Methods According to the procedure described by the NCCLS approved guideline EP9-A and improving the method of sample collection,we took 8 patient mixing samples each day to analyze all comparison tests on 11 auto-chemistry analyzer within following 5 days.The duplicates were assessed within the same run.The coefficient of correlation and average bias% were calculated,and the system errors at medical decided levels were assessed.Results Taking ALT as an example,the coefficients of correlation were between 0.994-1.000,and the average bias% were between-0.460%-4.927%,SE at 40U/L was-1.510-1.834 and SE at 300 U/L was-3.101-9.188.Conclusion In all tests that joined the comparison among the different instruments,28 tests were acceptable,2 tests were acceptable after modifying the coefficients,and AMY and LIP were not acceptable.

OBJECTIVETo determine the effect of cysteinyl receptor agonist leukotriene D(4) (LTD(4)) and its antagonists on rat primary neurons.

METHODSIn the primarily cultured rat cortical neurons, the neuron injury was evaluated by measuring intracellular calcium, 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyl tetrazolium bromide (MTT) reduction, and propidium iodide (PI) and Hoechst 33258 staining. The in vitro ischemic injury was induced by oxygen-glucose deprivation (OGD) for 1.5 h and reperfusion for 24 h.

RESULTLTD(4) (0.01-1 micromol/L) did not induce the elevation of intracellular calcium, neuron viability changes and neuron death. OGD-induced injury was not significantly ameliorated by the CysLT(1) receptor antagonists, pranlukast (0.2-10 micromol/L) and montelukast (0.2-10 micromol/L), as well as by the CysLT(1)/CysLT(2) receptor non-selective antagonist, BAY u9773 (0.02-1 micromol/L).

CONCLUSIONNeither agonist nor antagonists of cysteinyl receptors have the effects on the viability and ischemic-like injury in rat primary neurons.

Acetates ; pharmacology ; Animals ; Animals, Newborn ; Calcium ; metabolism ; Cell Hypoxia ; Cell Survival ; drug effects ; Cells, Cultured ; Cerebral Cortex ; cytology ; Chromones ; pharmacology ; Glucose ; pharmacology ; Leukotriene Antagonists ; pharmacology ; Leukotriene D4 ; pharmacology ; Neurons ; cytology ; drug effects ; metabolism ; Quinolines ; pharmacology ; Rats ; Receptors, Leukotriene ; agonists

OBJECTIVETo determine whether homeostatic conditions (pH, glycine or ion concentration) affect the protective effects of edaravone on ischemic injury in rat cortical neurons.

METHODSIn cultured rat cortical neurons, the compositions in the experimental solutions were changed to mimic the disturbance of homeostasis after cerebral ischemia. In vitro ischemic injury was induced by oxygen-glucose deprivation (OGD) for 3 h and reperfusion for 12 h, and the neuron injury was evaluated by 3-(4, 5-dimethylthiazol-2yl)-2, 5-diphenyl tetrazolium bromide (MTT) reduction assay and lactate dehydrogenase (LDH) release. Effect of edaravone on OGD injury was observed in different experimental solutions.

RESULTIn weak alkalified solution (pH 7.8) or the solution containing glycine (10 micromol/L), OGD injury became more serious; but in weak acidic (pH 6.5) or higher Mg(2+) (1.8 mmol/L) solutions, OGD injury was attenuated. Edaravone (1 micromol/L) reversed the injury in the solutions with pH 6.1,7.4 and 7.8 or the solution containing glycine, but did not show protective effect in the solution with pH 6.5 and the higher Mg(2+) or lower Ca(2+) solution.

CONCLUSIONThe changes of homeostatic conditions affect the severity of ischemic injury of neurons and the protective effect of edaravone.

Animals ; Animals, Newborn ; Antipyrine ; analogs & derivatives ; pharmacology ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; cytology ; Glycine ; pharmacology ; Homeostasis ; Hydrogen-Ion Concentration ; Magnesium ; pharmacology ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Rats ; Reperfusion Injury ; prevention & control

OBJECTIVETo determine the effect of histamine on N-methyl-D-aspartate (NMDA) induced neuron death and to elucidate its mechanism.

METHODSThe primary cortical cell culture was adopted. Neuron morphology and MTT assay were used to evaluate the drugs effects.

RESULTHistamine at doses of 10(-4) 10(-6) 10(-7) 10(-8) mol/L reversed the neuron death induced by NMDA (50 micromol/L) for 3 h. The protection of histamine peaked at doses of 10(-4) mol/L and 10(-7)mol/L. The effect of histamine of 10(-7) mol/L was reversed only by cimetidine an H(2)receptor antagonist. However, the effect of histamine of 10(-4) mol/L was reversed only by pyrilamine but not cimetidine.

CONCLUSIONHistamine could reduce neuron death induced by NMDA; its protection at a low dose might be mediated by H(2)receptor, and at a high dose by H(1)receptor.

Animals ; Cell Death ; drug effects ; Cells, Cultured ; Dose-Response Relationship, Drug ; Histamine ; pharmacology ; N-Methylaspartate ; toxicity ; Neurons ; drug effects ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Histamine H1 ; physiology ; Receptors, Histamine H2 ; physiology

OBJECTIVETo establish a simpler and more accurate method for evaluating in vitro ischemic injury and neuroprotective effects of drugs through improving experimental instrument and quantitative index in mouse brain slices.

METHODSAn incubation instrument was developed and its application tested. 2,3,5-triphenyltetrazolium chloride (TTC) was used as a substrate to biosynthesize formazan standard in mouse brain slices, and formazan was isolated, purified and identified. Ischemic injury of mouse brain slices was induced by oxygen/glucose deprivation (OGD), the produced formazan from TTC in the cortex and striatum was measured at 490 nm spectrophotometrically. Edaravone and ONO-1078 were added into the incubation medium to observe their neuroprotective effects.

RESULTThe incubation instrument worked well for incubating brain slices and obtaining stable results efficiently. Standard formazan was biosynthesized and purified with a purity of 99.3%, and showed a linear range of 0.05 - 1 mg/ml in absorbance at 490 nm (r=0.9997). OGD decreased formazan production in the cortex and striatum in a duration-dependent manner. Edaravone (0.01 to 1 micromol/L) recovered OGD-induced decrease of formazan production, but ONO-1078 showed no effect.

CONCLUSIONThe incubation instrument and quantitative measurement of formazan developed in this study are efficient,accurate and simple for evaluating ischemic injury and neuroprotection,which can be used in screening of neuroprotective drugs in vitro.

Alprostadil ; analogs & derivatives ; pharmacology ; Animals ; Antipyrine ; analogs & derivatives ; pharmacology ; Brain Ischemia ; diagnosis ; drug therapy ; Formazans ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Neuroprotective Agents ; pharmacology ; Staining and Labeling ; Tetrazolium Salts ; metabolism

OBJECTIVETo determine the protective effect of monosialoganglionside (GM1) and evaluate the influence of GM1 on expression of N-methyl-D-aspartate receptor subunit 1 (NMDAR1) in Sprague-Dawley (SD) rats with focal cerebral ischemia-reperfusion (I/R).

METHODSLeft middle cerebral artery (MCA) was occluded by an intraluminal suture for 1 h and the brain was reperfused for 72 h in SD rats when infarct volume was measured, GM1 (10 mg/kg) was given ip (intraperitoneally) at 5 min (group A), 1 h (group B) and 2 h (group C) after MCA occlusion (MCAo). Expression of NMDAR1 was detected by Western blot at various time after reperfusion (4 h, 6 h, 24 h, 48 h and 72 h) in ischemic hemispheres of the rats with or without GM1 administered.

RESULTS(1) Adjusted relative infarct volumes of groups A and B were significantly smaller than that of group C and the control group (P<0.01 and P<0.05, respectively). (2) Expression level of NMDAR1 was temporally high at 6 h after reperfusion, and dipped below the normal level at 72 h after reperfusion. GM1 at 5 min after MCAo significantly suppressed the expression of NMDAR1 at 6 h after reperfusion (P<0.05 vs the control). At 72 h after reperfusion, the NMDAR1 expression level of rats treated with GM1 administered (at 5 min or 2 h after MCAo) was significantly higher than that of the control (P<0.05).

CONCLUSIONGM1 can time-dependently reduce infarct volume in rats with focal cerebral I/R partly through stabilizing the expression of NMDAR1.

Animals ; Brain Ischemia ; metabolism ; pathology ; prevention & control ; G(M1) Ganglioside ; pharmacology ; therapeutic use ; Gene Expression Regulation ; drug effects ; Male ; Middle Cerebral Artery ; surgery ; Neurons ; drug effects ; physiology ; Protein Subunits ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Treatment Outcome

This study was aimed to investigate the expression characteristics of two transcriptional factors in Ikaros family, Ikaros and Helios isoforms and their mechanism, as well as their correlation with clinical parameters, which play important roles in transcriptional regulation of hematopoiesis. Expression of Ikaros and Helios isoforms in a total of 163 patients with leukemia and correlations between Ikaros and Helios isoforms were analyzed by PCR. The results showed that different expression patters of Ikaros and Helios isoforms existed in leukemia patients, that is, Ikaros isoform (Ik-6) was predominantly expressed in acute lymphoblastic leukemia (ALL) with BCR/ABL fusion gene, while Helios isoform (He-i) was overexpressed in T-cell ALL patients. The results of cloning and sequencing demonstrated that the isoforms of Ikaros and Helios had different genetic alterations. The statistical correlation between these two isoforms not was found in this study, although interaction between Ikaros and Helios has been reported. It is concluded that although Ikaros and Helios belong to the same family with similar structure of zinc fingers, their isoforms have different expression profile, specific genetic alterations, and different clinical relevance in patients with leukemia. The connection and interaction between Ik-6 and He-i needs further research.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Gene Expression Profiling ; Humans ; Ikaros Transcription Factor ; genetics ; metabolism ; Leukemia ; genetics ; metabolism ; Male ; Middle Aged ; Protein Isoforms ; genetics ; metabolism ; Young Adult

OBJECTIVETo investigate the expression of anaplastic lymphoma kinase (ALK) gene fusion antibody in non-small cell lung cancer (NSCLC) and explore the clinicopathological significance.

METHODSUsing manual immunohistochemistry (IHC) with D5F3 rabbit monoclonal antibody, we detected the expression of ALK gene fusion protein in 519 cases of NSCLC. The relations of ALK fusion protein with the clinical characteristics of the patients and the histological classification of the tumors were analyzed. The expressions of ALK fusion protein were compared between surgical specimens and biopsy samples, and the consistency of manual IHC results was evaluated with the results of a fully automated IHC instrument and fluorescence in situ hybridization (FISH).

RESULTSThe positivity rate of ALK fusion protein was 11.37% (59/519) among the cases detected by manual IHC. The patients tended to have a young age of onset (P=0.048) and most of the tumors were adenocarcinoma. In the surgical specimens, ALK fusion protein was expressed mostly in invasive mucinous adenocarcinoma (P<0.01), and it was a high risk factor of lymph node metastasis [OR=2.188(95%C.I:1.161-4.122)]. No statistical difference was found in the test results of manual IHC between surgical specimens and biopsy samples. The results by manual IHC suggesting a strong expression were consistent with the results by automated IHC and FISH.

CONCLUSIONManual IHC can be reliable for screening ALK fusion arrangement in patients with NSCLC.

Adenocarcinoma ; genetics ; Antibodies ; Carcinoma, Non-Small-Cell Lung ; genetics ; Gene Fusion ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Lung Neoplasms ; genetics ; Receptor Protein-Tyrosine Kinases ; genetics ; immunology

AIMTo investigate the protective effect of monosialoganglioside (GM1) on injury induced by oxygen glucose deprivation/reperfusion (OGD/Rep) in rat hippocampal slices.

METHODSThe protective effects of GM1 on hippocampal slices after OGD/Rep were observed by detecting the light transmittance (LT) changes of rat hippocampal slices and 2, 3, 5-triphenyltetrazolium chloride (TTC) staining of rat hippocampal slices.

RESULTS(1) In four groups treated with 0 (control), 0.1, 1.0, 10 micromol/L GM1, the peak of light transmittance (LT) in the slices treated with 1.0 micromol/L GM1 was significantly lower than that of the control and the group treated with 0.10 micromol/L GM1 (P < 0.01, ANOVA), while the peak of LT in the slices treated with 10.0 micromol/L GM1 was significantly lower than that of the other groups (P < 0.01, ANOVA). The time to reach the peak of LT in four groups was significantly different from each other (P < 0.05, Kruskal-Wallis test). The time to reach the peak of LT in the group treated with 1 micromol/L GM1 was the significantly longer than that in the control (P < 0.01, Mann-Whitney U test). (2) There was characteristic dose-response relationship between GM1 and TTC staining of rat hippocampal slices. In the five groups, treated with 0 (control), 0.01, 0.1, 1.0, 10 micromol/L GM1 respectively, TTC staining in the group treated with 1 micromol/L GM1 was the deepest (P < 0.05 vs. control, 0.01 and 0.1 micromol/L GM1 group, ANOVA), and the next was in the group treated with 10 micromol/L GM1 (P < 0.05 vs. control and 0.01 micromol/L GM1 group, ANOVA).

CONCLUSIONGM1 could protect injury induced by OGD/Rep in rat hippocampal slices effectively in vitro.

Animals ; G(M1) Ganglioside ; pharmacology ; Glucose ; deficiency ; Hippocampus ; metabolism ; Hypoxia ; metabolism ; In Vitro Techniques ; Male ; Oxygen ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury