Objective:To explore the influential factors of cognitive change among Chinese oldest-old.Method: Three waves of data from the Chinese Longitudinal Healthy Longevity Survey(CLHLS)were analyzed with a two-level repeated measures model.Results:In baseline,the male had a higher mean MMSE score than the female(27.0?3.7/24.4?5.6,P

Objective To explore the influencing factors for the purity and viability of primary cultured cortical neurons of rat,and optimize the separating and cultivating conditions of the cortical neurons.Methods The primary cotical neurons were cultured in a serum-free culture system of B27-supplemented neurobasal medium.The differences in purity and viability of primary cultured neurons between embr-yonic rat group and newborn rat [postnatal 24 h and 5 d] group were evaluated by morphology,immunocytochemistry of neuron-specific enolase(NSE) and trypan blue staining.The changes of neurons purity and viability in different trypsin digestion time(0 min,5 min and 15 min) at different environment temperatures(20 ℃ and 30 ℃) were assessed by immunocytochemistry and trypan blue staining.Results The primary cultured neurons from fetal and newborn rats grew well.There was no significant difference in embryonic rat and postnatal 1 d newborn rat group[(91.30?1.03)%,(89.50?1.78)% respectively in purity;and(98.20?0.58)%,(97.10?0.98)% respectively in viability].The neurons from 5-days newborn rat were inferior to that from fetal and 1-day newborn rat in purity and viability[(82.00?1.25)% and(92.87?1.56)% respectively].Shortening operation time and adjusting digestion time according to the environment temperatures could improve neuronal viability:A digestion for 15min at the environment temperature of 20 ℃ or for 5 min at 30 ℃ could acquired cells with higher viability [(98.20?0.58)% and(96.70?0.64)% respectively].Conclusions It is an easy,practical choice to culture priamry cortical neurons from postnatal 1 d newborn rats.Optimizing the separating and cultivating condition(environment temperatures,digest time,et al.) will improve the neurons purity and viability.

Objectives To investigate dynamic pathological features and virus distribution in the liver with a murine severe acute respiratory syndrome(SARS)model injected with murine hepati- tis virus 3(MHV-3)through trachea.As a representative of host genes,mouse fgl2(mfgl2)pro- thrombinase gene expression and its clinical significance were discussed in SARS associated liver dam- ages.Methods The Balb/cJ mice were infected with 100 PFU of MHV-3 through trachea and Balb/ cJ mice injected with saline were served as control.Survival rate,pathological features in organs and liver function were observed.Virus titers in different organs were determined on monolayer of L2 cells by a standard plaque assay.Virus distribution and cellular localization were studied by in situ hy- bridization.Both mfgl2 and fibrin expressions were examined in the liver by in situ hybridization and immunohistochemistry to investigate the role of mfgl2 in the liver impairment.Results Mice infected with MHV-3 through trachea developed multiple organs damages and died within 5 days,while all mice in control group survived with no histopathological changes.Infected liver tissues showed wide- spread cloudy swelling,prominent ballooning degeneration with mild lymphocytic infiltration in the portal area.Dot and zonal hepatocellular necrosis could be found occasionally.The lungs showed typi- cal interstitial pneumonia and hyaline membranes formation.Other histological changes also could be found in other organs examined.MHV-3 virus replication was identified in all organs observed.The liver function was injured,mfgl2 expression were evidenced mainly in the necrosis areas with fibrin deposition around the necrosis areas.Conclusions Pathological changes of the liver in this murine SARS model can mimic the liver impairment characteristics of SARS in human.In addition to the physical damage induced by the virus,the up-regulation of novel gene mfgl2 in the liver in association with fibrin deposition may play a vital role in the development of SARS associated liver damages.

Insulin receptor substrate(IRS)-1 and IRS-2 expression levels of liver tissues and skeletal muscle in intrauterine growth retarded(IUGR)rats were investigated by RT-PCR and immunohistochemistry.An IUGR animal model was established by maternal nutrition restriction during pregnancy.IRS-2 expression level of liver tissue and IRS-1 expression level of skeletal muscle in IUGR rats at 0 and 3 weeks old were significantly lower than those in normal rats at the same age respectively,and insulin resistance was induced in IUGR,and these findings might be the molecular mechanisms susceptible to metabolic syndrome in IUGR rats.

0.05).The concentration of urinary KYNA,metabolite of the KYN,was significantly lower in SHRs compared to WKYs(7.8?1.8 vs 19.9?3.5 ?mol/24 h P=0.013).Both KAT activity in renal cortex and KYNA content in urine were negatively correlated to blood pressure(r=-0.418,P=0.023;r=-0.723,P=0.001).Conclusion The declined activity of KAT in renal cortex and the deficiency of KYNA concentration in urinary may affect blood pressure regulation in SHR by renal metabolite of the KYN.

Objective To measure the renal tissue texture or flexibility with virtual touch quantization (VTQ) and to tentatively examine its clinical application in patients with chronic kidney disease(CKD).Methods A total of 750 patients (1500 kidneys) were performed with VTQ,including 400 cases in the control group,and 350 cases in the CKD group.A conventional ultrasound examination (two-dimensional,color Doppler) were first taken,and then the shear wave velocity (Vs) was measured which reflected the textural elastic.Results In both groups the Vs was the highest in renal cortex with significant difference (P<0.05); renal cortical region Vs in CKD group was lower than those in control group (P<0.05),while Vs of renal medulla and renal sinus had no significant difference in the two groups.The severity of renal dysfunction was increased along with a Vs decrease of renal cortex.Conclusion VTQ is helpful to assess renal function of patients with CKD.

OBJECTIVETo investigate the effects of inward rectifier potassium channel blockers (BaCl2, CsCl) on the functions of endothelial progenitor cells (EPCs).

METHODSDensity gradient centrifugation-isolated rat hone marrow mononuclear cells were cultured in vitro. EPCs were harvested and seeded on six culture dish when cells grew to 3-5 passages. Before testing the EPCs were synchronized with M199, which contain 2% fetal calf serum. In the end, EPCs were treated with different intervention. The experiment mainly included two parts: (1) BaCl2 (100 micromol/L) and free BaC2 of Tyrodes solution; (2) CsCl (1 mmol/L) and control. Cell pretreated with blockers above mentioned for 12 h, then the gene expression of stromal cell-derived factor-1 (SDF-1), epoprotenol (PGI2) were assessed, beyond that the ability of adhesion, migration were assayed with different tests. In addition, the medium was collected when EPCs were treated for 3 days. The levels of SDF-1 were measured by sandwich enzyme-linked immunosorbent assay (ELISA). Going even further, EPCs were treated with the signal pathway blockers in advance, after repeat the above steps, in order to analyze the change of SDF-1 and then discuss its mechanism.

RESULTSCompared with control group, BaCl2, CsCl could increase EPC adhesion and migration to same extent. Moreover, the gene expression of SDF-1, PGI2 was significantly up-regulated and the production of SDF-1 increased evidently. Furthermore, the mechanism of SDF-1 secretion increasing mainly was associated with eNOS signaling pathways.

CONCLUSIONBa2+ and Cs+ play important roles in increasing EPCs functions, such as adhesion, migration and secretion.

Animals ; Barium Compounds ; pharmacology ; Cells, Cultured ; Cesium ; pharmacology ; Chemokine CXCL12 ; metabolism ; Chlorides ; pharmacology ; Endothelial Cells ; cytology ; Enzyme-Linked Immunosorbent Assay ; Potassium Channels, Inwardly Rectifying ; antagonists & inhibitors ; physiology ; Rats ; Stem Cells ; cytology

Objective To explore the relationship between pulse wave velocity(PWV)and blood pressure classification and cardiovascular risk factors stratification in patients with essential hypertension. Methods Totally,390 patients with essential hypertension were recruited in the study and their carotid- femoral PWVs were recorded by an automatic pulse wave velocity measurement system.The patients were stratified by their cardiovascular risk factors.PWVs were compared between patients with varied blood pressures or with and without cardiovascular risk factors.Results There was no significant difference in PWV between patients of phase 1 and phase 2 hypertension,[(10.8?1.7)m/s vs.(11.9?1.2)m/s,P =0.398].PWV in patients with more than one risk factor was significantly higher than that in those with hypertension alone,[(10.4?1.4)m/s vs.(11.7?1.4)m/s,P=0.018].Conclusions PWV in hypertensive patients with one or more than one other risk factors was significantly higher than that in those without other risk factors.Cardiovascular risk factor might play a more important role in arterial stiffness.

To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice, ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units (PFU) of MHV-3 intraperitoneally. The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin (HE) staining method from 2 days post MHV-3 infection. The ratios of T cell subsets including CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4-CD8-, CD3+CD4+CD25+, CD3+CD4+CD25- and CD3+CD4-CD25+ T lymphocyte of total T lymphocytes in blood, spleen and liver were examined at 0, 2, 4, 6,8, 10, 12, 15, 20, 25, 30, 40 days post MHV-3 infection by flow cytosorting. We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection. The double negative T cell (DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice, and CD3+CD4+CD8-, CD3+CD4-CD8+, CD3+CD4+CD25- and CD3+CD4-CD25+ T cell ratios decreased accordingly. In conclusion, the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence. Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection. Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.

Objective:To prepare the polyclonal antibodies against advanced oxidation protein products (AOPP),and to provide an effective agent for research on the pathogenesis of AOPP and assess exactly the relationship between AOPP and relative diseases.Methods:AOPP-rabbit serum albumin (AOPP-RSA) was prepared by treating RSA with hypochloric acid.The rabbit anti-AOPP-RSA polyclonal antibodies were generated and purified by affinity chromatography. The titers and the specificity of the antibodies were measured by ELISA.The plasma AOPP and the localization of AOPP in nephridial tissues of some patients with chronic kidney disease (CKD) were determined using rabbit anti-AOPP-RSA.Results:Titers of the antibodies were 10-6.Purified antibodies reacted specifically with oxidized albumin from different genus,but could not react with normal albumin and glycosylated albumin.The high level of AOPP in plasma from CKD patients was confirmed by Western blot.The antibodies could be used to immunostain AOPP deposition in different regions of kidney tissues from both CKD patients and CKD rat models.Conclusion:We successfully generate rabbit anti-AOPP polyclonal antibodies with high titers and striking specificity.The presence of plasma AOPP and localizations of AOPP in kidney tissues of CKD patients can be demonstrated using the antibodies.The development of anti-AOPP polyclonal antibodies may provide a new tool to explore the pathogensis of AOPP and assess exactly the relationship between AOPP and relative diseases.