Objective To investigate the effects of deletion of adenosine A2A receptors on peripheral leukocytes on cerebral white matter lesions induced by chronic cerebral hypoperfusion.Methods Twenty-four wild type(WT) male mice were given a ? irradiation of 12.5Gy,followed by receiving bone marrow cells tail vein from female A2A receptor knocked out(KO) mice via tail vein,were assigned as KO→WT group,while those received bone marrow cells from WT female mice were assigned as WT→WT group(n=20).The efficiency of reconstitution of bone marrow cells in recipient mice was assessed 7 weeks after transplantation by PCR and immunofluorescent technique.Then,the recipient mice were subjected to bilateral common carotid artery stenosis with internal diameter of 0.18mm by external banding using microcoils at 8 weeks after transplantation.On 7d,14d and 30d after the surgery,corpus callosum,fiber bundles of Caudoputamen and optic tract were harvested from the cerebral white matter,and stained with Kluver-Barrera staining for observing the changes in nerve fibers,and with GFAP and CD11b immunohistochemistry staining for observing the proliferation of microglia and astrocytes.Results At 7 weeks after successful transplantation,the genotype of sex chromosome in peripheral leukocytes of the male recipient mice was changed into female pattern.The expression rate of A2A receptor was 9.73%?2.05% in KO→WT group and 93.82%?11.24% in WT→WT group,with significant difference between the two groups(P

Objective To observe behavioral characteristics and pathologic changes of chronic cerebral hypoperfusion in mice. MethodsTotally 62 adult C57BL/6 mice were subjected to either sham-operation (n=31) or bilateral common carotid artery stenosis using external microcoils with an inner diameter of 0.18 mm(n=31). At 30 d after the stenosis, the animals of the 2 groups (8 mice for each group) underwent behavioral test of 8-arm radial maze. In 7, 14 and 30 d, the rest mice were sacrificed for their brain tissue samples which were examined with Kluver-Barrera staining and immunohistochemical assay for markers of microglia and astroglia, glial fibrillary acidic protein (GFAP) and CD11b. In 14 d after the model establishment, Evans blue dye extravasation test was performed for the blood-brain barrier function. ResultsThe model group made significantly more errors than sham-operated group in 8-arm radial maze test at 30 d after the surgery. White matter lesions occurred and the proliferation of activated microglia and astroglia were observed in white matter in model mice after 14 and 30 d bilateral common carotid artery stenosis. The disruption of blood-brain barrier function of model mice was indicated in the evans blue extravasation test at 14 d after bilateral common carotid artery stenosis. ConclusionCognitive impairment, white matter lesions and glial activation are successfully induced after bilateral common carotid artery stenosis in mice model of chronic cerebral hypoperfusion.

Objective To investigate the effect of adenosine A2A receptor deficiency on the ischemic neuronal injury and its potential mechanism.Methods Transient(2 h)cerebral ischemia was induced by the middle cerebral artery occlusion(MCAO)in mice.Adenosine A2A receptor knockout(A2ARKO)mice and their wild-type littermates(A2ARWT)were divided into 2-hour cerebral ischemia group,cerebral ischemia with 22-hour reperfusion group and cerebral ischemia with 46-hour reperfusion group.Cerebral infarction volume was measured by image analysis of brain sections stained with cresyl violet(CV).Brain water content was evaluated with the dry-wet weighing method.The expression of calbindin D-28k(CB)and aquaporin-4(AQP4)in ischemic brain was determined with immunohistochemical methods.Results The cerebral infarction volumes in 2-hour cerebral ischemia group,cerebral ischemia with 22-hour reperfusion group and cerebral ischemia with 46-hour reperfusion group of A2ARKO mice were lesser than those in the corresponding groups of A2ARWT mice.Compared with A2ARWT mice,A2ARKO mice had more CB,lesser AQP4 expressions and lesser brain water contents.Conclusion Adenosine A2A receptor deficiency exerts the protection against ischemic brain injury both in the acute phase and reperfusion phase,and attenuates brain edema caused by cerebral ischemia,which may be due to the inhibition of intracellular calcium overload and AQP4 expression.

Objective:To observe influence of alprostadil injection on contrast-induced nephropathy (CIN) in high risk patients after percutaneous coronary intervention (PCI).Methods: A total of 263 CIN high risk (CIN risk score ≥16 scores) patients were selected.According to random number table, patients were randomly divided into routine treatment group (n=121, received routine hydration therapy) and alprostadil group (n=142, received additional alprostadil injection based on routine treatment group).Serum creatinine (SCr), glomerular filtration rate(GFR), cystatin C (CysC) and β trace protein (β-TP) level before, 48h and 72h after PCI were measured and compared, and incidence rate of CIN, percentage of blood purification therapy and mortality were compared between two groups.Results: Compared with before PCI, there was significant rise in SCr level and significant reduction in GFR in both groups on 48h and 72h after PCI (P<0.01 all);Compared with routine treatment group, there were significant reductions in levels of SCr [72h: (190.04±28.92) μmol/L vs.(141.10±21.18) μmol/L], and significant rise in GFR [72h: (26.0±4.4) ml/min vs.(36.4±4.9) ml/min], and levels of CysC[72h: (1.75±0.74) mg/L vs.(1.47±0.55) mg/L] and β-TP [72h: (1.53±0.50) mg/L vs.(1.22±0.38) mg/L] significantly decreased in alprostadil group on 48h and 72h after PCI, P<0.05 or <0.01;there were significant reductions in incidence rate of CIN (30.6% vs.18.3%) and percentage of blood purification therapy (10.7% vs.3.5%) in alprostadil group, P=0.001, 0.045 respectively.There was no significant difference in mortality between two groups, P=0.728.Conclusion: Alprostadil injection can significantly improve kidney function, reduce incidence rate of CIN and percentage of blood purification therapy in CIN high risk patients after PCI, which is worth extending.

Objective To identify the effect of both high glucose and high insulin on miR-145 level.Methods The human vascular smooth muscle cells ( VSMCs) were cultured and the proliferation of VSMCs was induced by high glucose and high insulin medium.The samples were divided into 4 groups: normal group, high glu-cose group (25 mmol/L), high insulin group (300 mU/L), high glucose and high insulin group.The ex-pression of miR-145 in VSMCs was assayed by real-time PCR.Proliferation of VSMCs was determined by MTT method After 72 h cultivation.The migration of VSMCs was analyzed by cell scratch test .VSMCs in each group was transfected by miR-145 virus ( lentiviral vector ) .Proliferation and migration were assayed after 48 h transfection.Results The expression of miR-145 in VSMCs of other three groups was decreased ( P<0.05 ) , especially the expression in the high glucose and insulin group was the lowest ( P<0.01 ) .Prolifera-tion and migration of VSMCs was promoted by high glucose and/or high insulin medium.Under fluorescent, transfection rate of VSMCs was about 80%after 48 h transfection.Proliferation and migration of VSMCs in each group after transfection were significantly lower than before ( P<0.05 ) .Conclusions High glucose and high in-sulin could decrease the expression of miR-145 in VSMCs, The overexpression of miR-145 may inhibit the prolif-eration and migration of VSMCs .

Objective To explore the predictive value of combined ABCD2 scores and tracheal constriction resulted from internal carotid atherosclerosis (ICAS) in cerebral infarction within 7 d of onset of a transient ischemic attack (TIA). Methods The clinical data of 171 patients with TIA,admitted to our hospital from September 2008 to March 2011, were retrospectively analyzed. All patients were evaluated with ABCD2 scale and degrees of tracheal constriction resulted from ICAS were examined by Color Doppler ultrasound.And 7-d follow-up of patients was performed to select those patients appearing cerebral infarction; and then, the relationships between risk of post-TIA cerebral infarction and both ABCD2 scores and degrees of tracheal constriction were analyzed using chi-square test. Results Among 171 TIA patients,39 (22.8％) suffered cerebral infarction within 7 d of onset of TIA.Significant difference of risk of post-TIA cerebral infarction was noted in patients with different ABCD2 scores and degree of tracheal constriction (P＜0.05): the higher the ABCD2 scores/ the degrees of tracheal constriction, the higher incidence of cerebral infarction in TIA patients. Conclusion A rather high accuracy may be achieved with ABCD2 scale in the prediction of short-term risk of cerebral infarction after onset of TIA,and a higher accuracy may be expected by combination of ABCD2 scale with carotid atherosclerosis findings.

Solid-phase microextraction is a new technique of analysis. It has many merits and expanse foreground. A Review of the principle, recent development and applications of solid-phase microextraction is given, focusing on natural product analysis, especially on Chinese traditional medicine. Twenty-nine references are cited in the paper.
Animals ; Chromatography, Gas ; methods ; Chromatography, High Pressure Liquid ; methods ; Cotinine ; urine ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Food Analysis ; Humans ; Plants, Medicinal ; chemistry

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis (EP) is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells (PBMCs) derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris (PV) patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls (P<0.05). Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients (P<0.01). The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.

The effects of the feed attractants on Whitmania pigra were studied. The average weight of Wh. pigra were 5.0 g. Arginine was selected as feed attractants, xanthan gum was selected as feed substrate. The times of Wh. pigra going into the inducing room were recorded. The water temperature was 22-25 degrees C during the whole experiment. Arginine that had better inducing effect was chosen to carry on in the gradient experiment. The results showed that the best inducing effect was found when the added amount of arginine was 0.3%, which was close to the arginine content of the natural body fluid of Wh. Pigra and Bellamya purificata, 2.97 mg x g(-1).
Animal Feed ; analysis ; Animals ; Arginine ; analysis ; metabolism ; Body Weight ; Feeding Behavior ; Leeches ; growth & development ; physiology

This study was purposed to investigate the molecular mechanism of 4-1BBL reverse signals in the human acute monocytic leukemia cell line of U937. The U937 cell line was used as target cells, and stimulated by the mouse anti-human 4-1BBL monoclonal antibody 1F1. The nuclear translocation of NF-κB and the co-location of 4-1BBL and CD28i molecules in U937 cells were observed with confocal laser scanning microscopy. The protein and m-RNA expression levels of 4-1BBL and CD28i were detected by flow cytometry and RT-PCR respectively. The results showed that the significant nuclear translocation of NF-κB and co-localization of 4-1BBL and CD28i on membrane of U937 cells appeared after being stimulated by mAb1F1. It is concluded that the 4-1BBL reverse signals transduction mediating the growth of U937 cells relates with the nuclear translocation of NF-κB. CD28i may be involved in intracellular 4-1BBL reverse signaling pathways.
4-1BB Ligand ; immunology ; metabolism ; Antibodies, Monoclonal ; pharmacology ; CD28 Antigens ; metabolism ; Coculture Techniques ; Humans ; NF-kappa B ; genetics ; Signal Transduction ; U937 Cells