Objective To explore the relation between serum nesfatin-1 ,tumor necrosis factor (TNF)-αand insulin resistance. Methods A total of 105 subjects were enrolled in this study and divided into two groups:patients with newly diagnosed type 2 diabetes mellitus (T2DM group,n= 64)and normal controls (NC group,n= 41). The fasting serum nesfatin-1 and TNF-αlevels were measured by enzyme-linked immuno sorbent assay (ELISA). FPG,HbA1 c,TG,TC,and FIns were also tested. BMI, HOMA-IR,HOMA-β,and ISI were calculated. Results Serum nesfatin-1 and TNF-αlevels were significantly higher in T2DM group than in NC group. Multiple linear regression analysis showed that the most significant influencing factors for nesfatin-1 were TNF-αand ISI(β= 0. 005、-6. 847,P<0. 05). The most significant influencing factors for TNF-αwas HbA1 c (β= 26. 652,P<0. 01). Conclusion Serum nesfatin-1 and TNF-αare significantly elevated in patients with newly diagnosed T2DM,which may influence the glucolipid metabolism through the signal pathway of insulin and play a role in the pathogenesis of T2DM and IR.

BACKGROUND: The primary pathophysiology of Alzheimer disease (AD) is linked to β-amyloid (Aβ)protein. Neural progenitor cells (NPCs), which have the ability of multipotency, self-renewal and repair,have been detected in the central nerve system (CNS) of adult rat recently. But effective function of these neural progenitor cells are not seen in the AD brain ,which mechanism is unclear.It is unclear if Aβ1-40protein is compromised by the signal pathway of c-Jun N-termial kinase associated with the neurotoxicity to the progenitor cells on the cortex of embryonic rats.OBJECTIVE: To investigate the mechanism of c-Jun N-terminal kinase signal transduction pathway of Aβ1-40 protein, which has neuronal toxicity to progenitor cells(CPC)on the cortex of embryonic rats . To detect the neuroprotective effects of c-Jun N-termial kinase inhibitor (SP600125) against Aβ1-40-induced neuronal toxicity to the cortical progenitor cells on the cortex of embryonic rats.DESIGN: A randomized and controlled trial with cells as objects.SETTING: Department of Neurology, First Hospital Affiliated to China Medical UniversityMATERIALS: This experiment was carried out at the Central Laboratory,China Medical University from May to October 2005. Embryos at age of 14 days from Wistar rats were used in this experiment.METHODS: Cortical progenitor cells harvested from Wistar embryonic rats were cultured in vitro, passaged and identified. Embryonic rat cortical progenitor cells of rats with good growth state were randomly divided into 4groups:Aβ1-40 group (10 nmol/L Aβ1-40 in each well);SP600125+Aβ1-40group (10 μmol/L SP600125 for 30 minutes and then with 10 nmol/L Aβ1-40 in each well); SP600125 group ( 10 μmol/L SP600125 in each well); Normal saline group (same volume of normal saline). The incubated durations were 0,2 hours, 4 hours, 6 hours, 12 hours, 24 hours respectively,8 wells for each time point. The cell survival rate was measured by MTF assay (The concentration of cortical progenitor cells on the cortex was 1×10s L-1 in each group), the apoptosis rate was detected by flow cytometer (The concentration of cortical progenitor cells on the cortex was 1 ×1010 L-1in each group) and the expression of c-Jun N-termial kinase and p-c-Jun N-termial kinase, c-Jun,p-c-Jun were measured by Western Blot(The concentration of cortical progenitor cells on the cortex was 1×1013 L-1 in each group). t test was adopted for the comparison of difference in measurement data.sion of c-Jun N-terminal kinase, p-c-Jun N-termial kinase ,c-Jun and p-c-Jun of embryonic rat CPC .ture time in Aβ group and SP600125 +Aβ group, decreased obviously at 4hours; cellular survival rate in Aβ1-40 group was lower obviously than that in the other 3 groups at 0,2,4,6,12,24 hours (P ＜ 0.01); Cellular survival rate in SP60025 +Aβ1-40 group was lower obviously than that in SP600125 group and normal saline group at 2,4,6,12,24 hours (P ＜ 0.01);Compared with normal saline group, the difference of cell survival rate was not significant without time-dependent manner in SP600125 group (P＞ 0.05).amyloid protein group and SP600125 +Aβ group, increased obviously at 4hours; cell apoptosis rate in Aβ1-40 group was higher obviously than that of the other 3 groups at 0,2,4,6,12,24 hours(P ＜ 0.01); Cellular apoptosis rate in SP60025+Aβ1-40 group was higher obviously than that in the SP600125 group and normal saline group at 2,4,6,12,24 hours (P ＜ 0.01);Compared with normal saline group, the difference of cellular apoptosis rate was not significant without time-dependent manner in SP600125 group 12,24 hours without changes in Aβ1-40 group; the expression of p-c-Jun N-terminal kinase and p-c-Jun in Aβ1-40 group were seen at 0hour ,increased gradually, reached to the peak at hour 4 and decreased gradually.CONCLUSION: Aβ1-40 could inhibit the cell activity of CPC , reduce cellular survival rate and induce cellular apoptosis. c-Jun N-terminal kinase signal transduction pathway may mediate the Aβ1-40 inducd neurnal apoptosis in AD which may be one reason for unseen rescue mechanism in AD. SP600125 (c-Jun N-terminal kinase inhibitor) could inhibit the activation of c-Jun N-terminal kinase and c-Jun and protect the embryonic rats CPC from the Aβ1-40-induced neurotoxicity.

Objective:To prepare and identify the monoclonal antibody against hPPAR?2.Methods:cDNA of hPPAR?2 was obtained from pMD18-T/hPPAR?2.hPPAR?2 was expressed in prokaryocyte and purified successfully.The anti-hPPAR?2 monoclonal antibodies (mAbs) 1B4,3H2,3H10,10D6 and 10D8 were produced by immunization with purified hPPAR?2.The specificity of the antibodies was identified by ELISA,Western blot assay and immunocytochemistry.Results:Five novel murine monoclonal antibodies only against hPPAR?2 were harvested,and they were all specific to hPPAR?2.Conclusion:The antibodies obtained provide more useful tools for following research and settling the basis for screening new drugs and mechanism.

Objective To observe the effects of Tangshenjiaonang (TSJN) on biochemistry, renal function and hemorheology of alloxan and achromycin induced diabetic nephropathy rats. Methods The diabetic nephropathy rats induced by alloxan and achromycin were divided into 5 experimental groups. compared with control group, to observe the changes of the indicators shown about. Results TSJN can significantly decrease the level of SCr, BUN, CCr, FIB, plasma viscosity and UAER (P

0.05). After treating, the accumulating points of syndrome of the treatment group dropped more, the treatment group was superior to the control group. Conclusions The curative effects of Huoxin capsule on angina of coronary arheroselerosis heart disease is exact. It can take effect quickly and improve the whole symptom of the patients noticeably.

Diabetes Mellitus, Type 2 ; complications ; Diagnosis, Differential ; Follow-Up Studies ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Myelinolysis, Central Pontine ; complications ; diagnosis ; Prognosis

BACKGROUND: Techniques in the focal cerebral ischemia model in mice have been well developed in some foreign countries. However, rats are commonly used in cerebral ischemia studies due to resource availability in China.OBJECTIVE: To establish reliable and reproducible focal cerebral ischemia reperfusion model in mouse to explore the molecular mechanism of cerebral ischemia reperfusion process in genetic level.DESIGN: Randomized controlled study.SETTING: Fundamental Medical Research Institute of Chengde Medical College. MATERIALS: The experiment was carried out in Fundamental Medical Research Institute of Chengde Medical College from September 2002 to May 2003. Totally 30 Kunming mice (25 to 30 g), provided by Experimental Animal Center, Chengde Medical College, were randomly grouped as follows: control group, 6, 3-hour occlusion and 18-hour, 24-hour reperfusion, with 6 mice in each group respectively.INTERVENTIONS: Nylon thread (qb0.128 mm) was soaked in paraffin wax and inserted into internal carotid artery in mouse to reversibly occlude the middle cerebral at the right side after 3 hours, then was removed, and reperfusion was performed in 18, 24 hours, respectively.MAIN OUTCOME MEASURES: The infarct volume was observed and neurological deficit scored was analysis.of mice in operation group was shown as head turning to left, adduction of left forelimb, internal rotation of shoulder, decrease of muscular tension of left forelimb, unable to straighten completely during tail lifted. At independent activity, mice turned to left as the left hidlegs as the center of a circle; under the abdomen, left forelimb was crossed with right forelimb as scissor shape. Palsy of left limbs was observed obviously, especially forelimb.tion was shown clearly with TTC. The infarct volumes were (13±2.3) mm3 and (16±4.5) mm3 for 18 hours and 24 hours reperfusion and 3-hour ischemia, respectively. Pathological changes were observed under light microscope after ischemia. The infarct volume increased as the increasing of reperfusion time, including 3-hour blocking and 18-hour and 24-hour reperfusion.CONCLUSION: Using suture method to establish focal cerebral ischemia reperfusion model in mice, it is less invasion and trauma without requiring skull opening. Ischemia position is relatively fixed, thus, ischemia reperfusion time can be controlled precisely. It is also an ideal animal model to study pathological changes in cerebral vascular diseases and an efficient tool to study the therapeutic effect of medicine after surgery.

Background Hypoxia is the main factor of retinal neovascularization and is closely associated with retinal ganglial cells (RGCs) degeneration.However, the study of retinal neural tissue lesions is rare.Objective This study was to investigate the influence of hypoxia environment on the expression of annexin A2 (ANXA2) in mouse RGC-5 cells and explore the mechanism of RGCs damage induced by hypoxia.Methods Immortalized mouse RGC-5 cells were cultured in high glucose DMEM with 10％ fetal bovine serum.The cells were identified by detecting the expression of Thy-1 ,a specific biomarker of RGCs.CoCl2 was added into the medium at the final concentrations of 50,100,200 and 300 μmol/L, and the cells without CoCl2 served as the control group.Cell viability (absorbance) was assayed by cell counting kit-8 (CCK-8) method in 12,24 and 48 hours after addition of CoCl2.The hypoxic cell models were established in DMEM with 100 μmol/L CoCl2 and divided into the hypoxic 3-hour group, hypoxic 6-hour group, hypoxic 12-hour group and hypoxic 24-hour group,with the normal cultured cells as the normal control group.Apoptotic cells were determined by using hoechst 33342 stain.The expression levels of ANXA2 mRNA and protein in the cells were detected by real-time quantification PCR and Western blot,respectively.The expression and location of ANXA2 in the cells were examined by using immunofluorescence technique.Results The cultured cells grew well and showed the fusiform and polygonal shape,with positive expression of Thy-1 protein.Compared with the normal control group, the viabilities of the cells were insignificantly changed in the 50 μ mol/L CoCl2 group and 100 μmol/L CoCl2 group (all at P＞0.05) ,but the cell viabilities were significantly reduced in the 200 μμmol/L CoCl2 group and 300 μmol/L CoCl2 group in various time points (all at P＜0.05).Hoechst 33342 staining showed that the apoptotic cells with nuclear condensation and high green fluorescence intensity were obtained in the hypoxia groups.The relative expression levels of ANXA2 mRNA were significantly lower in the hypoxic groups than those in the normal control group (all at P ＜ 0.05).The relative expression levels of ANXA2 protein were significantly lower in the hypoxia 3-,6-, 12-and 24-hour group than those in the normal control group (all at P＜ 0.05).Apoptotic cells were seen in the hypoxic 3-hour group, hypoxic 6-hour group, hypoxic 12-hour group and hypoxic 24-hour group compared with the normal control group, showing the bright blue fluorescence in cellular nucleus for hoechst 33342.The relative expressing levels of ANXA2 mRNA in the cells were 0.80±0.14,0.67±0.33, 0.49±0.17 and 0.39±0.02 in the hypoxic 3-hour group,hypoxic 6-hour group, hypoxic 12-hour group and hypoxic 24-hour group, which were significantly declined in comparison with the normal control group, with a statistically difference among the groups (F=434.354, P =0.000).The relative expression values of ANXA2 protein were 0.552 6±0.012 3,0.425 9± 0.033 4,0.344 9 ± 0.017 8 and 0.382 7 ± 0.022 1 in the hypoxic 3-hour group, hypoxic 6-hour group, hypoxic 12-hour group and hypoxic 24-hour group,which were remarkably lower than 0.602 1 ±0.001 4 in the normal control group, showing considerably difference among the groups (F =3.057, P =0.000).ANXA2 proteins were highly expressed in the cellular nucleus and less expressed in the cell membrane and cytoplasm in the normal cells.Compared with the normal control group, the ANXA2 protein showed weak expression in the hypoxia group and primarily in the cytoplasm.Conclusions The expression of ANXA2 down-regulates in hypoxic mouse RGC-5 cells,which may participate in the apoptosis process of RGCs in high glucose environment.

Atherosclerosis is the most important cause of ischemic stroke. microRNAs can play an important role in the lipid metabolism, vascular inflammatory response, angiogenesis, as wel as smooth muscle cel proliferation and phenotypic conversion, etc. by regulating the functions of vascular endothelial cel s, vascular smooth muscle cel s, and mononuclear macrophages. It is closely associated with the occurrence and development of atherosclerosis. This article mainly reviews the regulatory effect of microRNAs on lipid metabolism and vascular inflammatory response in pathogenesis of atherosclerosis.

Objective To evaluate the role of SPECT cerebral perfusion imaging in assessing the stent implantation for cerebral artery stenosis.Methods A total of 35 patients (31 males,4 females,average age (63.9±10.8)years) with cerebral artery stenosis confirmed by DSA for cerebral artery stent implantation were retrospectively analyzed.99Tcm-ECD cerebral perfusion imaging was performed for all patients before and after stent implantation.The images were realigned and normalized by SPM 2.0 and then analyzed by Brain Search software for quantitative analysis.The brain was automatically separated to 210 functional areas according to Talarich map.The normalized averaged counts (NAC) of each area were calculated and compared with the data of 28 health controls (8 males,20 females,average age (35.8± 9.4) years).Less than 1.96s was defined as low perfusion lesions.The NAC values before and after stent implantation were compared for classifying improved from non-improved group.The mean number of lesions and Essen stroke risk score (ESRS) were analyzed between the two groups.The mean number of lesions and postoperative improvement rate of the internal carotid artery (ICA) occlusion and stenosis were compared.Paired rank sum test,two-sample t test,two-sample rank sum test and x2 test were used for statistical analysis.Results In 35 patients with low perfusion areas,20 were significantly improved after stent implantation.The mean number of lesions in the improved group (34.05± 14.41)was significantly higher than that in the non-improved group (22.93±17.24; t=2.067,P＜0.05).The mean ESRS of the improved patients (14.8)was significantly lower than that of the non-improved patients (22.3,Z=2.24,P＜0.05).The improvement rate of 28 cases with ICA stent implantation was (60.7％,17/28)higher than that of 7 cases with middle cerebral artery (MCA) stent implantation (3/7; P＞0.05).The mean number of the ICA occlusion lesions (34.36± 14.31)was higher than that of the ICA stenosis lesions(31.35± 16.37),but the difference was not statistically significant(t=0.498,P＞0.05).The improvement rate of the ICA occlusion was higher than that of the ICA stenosis (7/11 vs 10/17),but the difference was not statistically significant (P＞0.05).Conclusion SPECT cerebral perfusion imaging and its quantitative analysis can evaluate the low perfusion lesions before stent implantation and predict the perfusion improvement after stent implantation.