Objective To research the estrogen-like activity of the DDTs and the extract of vehicle exhaust by recombinant yeast system.Methods Being added into the yeast cell culture fluid,the objects reached the final concentration of 10-9～10-15 mol/L for 17?-estradiol,10-4～10-9 mol/L for DDTs(p,p'-DDT,o,p'-DDT,p,p'-DDD,p,p'-DDE),0.25～8.00 ml/ml for the extract of vehicle exhaust,then the cultivation was performed for 4 hours,the quantitative assay of ?-galactosidase activity accounts for the estrogenlike effect of the samples was carried out.Results DDTs were a group of environmental estrogens hormones,which could integrate estrogen receptors as the stimulators do and express the estrogen-like effect.Being tested separately,the DDTs showed the estrogen-like activity,which was stronger for o,p'-DDT and weaker for p,p'-DDE.The?-galactosidase activity of gasoline-fueled vehicle exhaust at the final concentration of 2.00～4.00 ml/ml was significantly different from that of the DMSO(P

Rat brain kinesin is a conventional kinesin that uses the energy from ATP hydrolysis to walk along the microtubule progressively. Studying how the chemical energy in ATP is utilized for mechanical movement is important to understand this moving function. The monomeric motor domain, rK354, was crystallized. An ATP analog, AMPPNP, was soaked in the active site. Comparing the complex structure of rK354 x AMPPNP and that of rK354ADP, a hypothesis is proposed that Glu237 in the Switch II region sensors the presence of gamma-phosphate and transfers the signal to the microtubule binding region.
Adenosine Triphosphate ; metabolism ; Adenylyl Imidodiphosphate ; metabolism ; Animals ; Brain ; metabolism ; Catalytic Domain ; Crystallography ; Hydrolysis ; Kinesin ; metabolism ; Microtubules ; metabolism ; Phosphates ; Protein Binding ; Rats

Kinesin is a motor protein that uses the energy from ATP hydrolysis to move along the microtubule system. To investigate how the chemical energy stored in ATP is converted to mechanical movement, the corresponding N-terminal region of rat brain kinesin was expressed in BL21-Codon Plus (DE3)-RP competent cells. After SP-cation exchange chromatography and size exclusion chromatography, the protein yield reached 10 mg/L culture with the purity above 95%. The purified protein had ATPase activity and specifically reacted with the kinesin antibody in the Western blotting analysis. The purified kinesin was crystallized under the following condition: 1.7 mol/L (NH4)2SO4, 500 mmol/L NaCl, 20% glycerol. The kinesin crystal can diffract up to 2.0 angstroms resolution.
Animals ; Brain ; enzymology ; Crystallization ; Hydrolysis ; Kinesin ; chemistry ; isolation & purification ; Rats

Objective:To explore the association between the changes in ideal cardiovascular health (CVH) indices and the renal function.Methods:The retrospective cohort study consisted of 837 Chinese individuals from Department of Health Management in the Third Xiangya Hospital.The general information,anthropometry and blood biochemistry were obtained for all subjects from 2008 to 2014,respectively.The 6-year changes of 7 ideal CVH metrics defined by American Heart Association and the estimated glomerular filtration rate (eGFR) were calculated.Linear regression analysis was used to study the association between the changes in the ideal CVH metrics and eGFR.Covariance analysis was used to compare the levels of changes in eGFR stratified by variations of the ideal CVH metrics.Results:After 6 years' follow-up,we did not find significant difference in ideal cardiovascular health indexes between 2014 and 2008 [(3.3±1.4) items vs.(3.2±1.3) items,respectively] (P＜0.05).However,the eGFR in 2014 was significantly lower than that in 2008 [(99.8±22.7) mL/(min.1.73 m2)vs (104.3±24.9) mL/(min.1.73 m2),respectively] (P＜0.01).After adjusting for age,sex,history of hypertension and diabetes,and history of drug usage (antihypertensive,cholesterol-lowering and blood glucose-lowering medication),the changes in ideal CVH metrics were positively associated with the changes in eGFR (β=0.701,P＜0.01).With the improved change in ideal CVH metrics from ≤-2 points,-1 point,0 point,1 point to ≥ 2 points,the magnitude of decline in eGFR was gradually decreased,with significant difference (F=21.71,P＜0.01).Conclusion:The changes in ideal CVH metrics are an independent risk factor for changes of eGFR.Positive changes in ideal CVH metrics exert a favorable effect on renal function evolution.

Objective To evaluate the effect of graded management of health in hypertensive populations.Methods Hypertensive adults who were detected in health checkup between January 2008 and June 2011 and with no severe liver or kidney disease,chronic consumptive disease and cancer were enrolled in this investigation.Those with low or modest hypertensive risk received grade 2health management,and those with high or extreme high risk got grade 3health management.At the end of the intervention,50％ participants were randomly selected in each year(1441 in 2008,1508 in 2009,1533 in 2010,and 1601 in 2011) to compare lifestyle modification,blood pressure,blood glucose and blood lipid profiles by using multiDunnett's t test and x2 analyses.Results In both grade 2 and 3 health management groups,compared with baseline,high-salt diet,current cigarette smoking,alcohol consumption and insufficient physical activity were significantly improved after intervention (x2 values were 154.27 and 81.16,52.43 and 9.37,61.40 and 12.13,46.54 and 12.05,respectively; all P ＜ 0.05).Blood glucose in grade 2 health management group and waistline in grade 3 health management group showed no significant change.Body weight,blood glucose,blood lipid profiles and serum creatinine of grade 2 health management group were significantly declined after intervention.In both groups,systolic and diastolic blood pressure were significantly decreased(F values were 13.12 and 56.86,56.86 and 239.89,respectively; all P ＜0.05),and those with normal blood pressure were increased from 28.8％ and 23.4％ to 37.9％ and 28.2％ in grade 2 and 3 health management groups(F values were 12.78 and 22.93,respectively ; all P ＜ 0.05).Conclusion These data suggest the beneficial effects of graded management of health on blood pressure control and reduced risks of cardiovascular disease.

Objective To study effects ofDanlong Xingnao Formula (DLXNF) on proliferation of neural stem cells (NSCs) and the expressions of Hes1 and Hes5 in sub ventricular zone (SVZ) in cerebral ischemia-reperfusion injury model rats; To explore the mechanism of promoting the proliferation of NSCsMethods Eighty male SD rats were randomly divided into sham-operation group, model group, edaravone group andDLXNF group. The focal cerebral ischemia reperfusion injury models were prepared by suture method, and 7 d after reperfusion, the SVZ brain tissue of ischemia side was taken. The proliferation of cells was detected by Brdu labeling fluorescence immunocytochemistry; Hes1, Hes5 mRNA and protein expressions were detected by fluorescence real-time quantitative PCR and Western blot method in each group.Results Compared with the sham-operation group, Brdu positive cell rate in other groups increased more obviously, and the expressions of Hes1, Hes5 mRNA and protein also increased significantly (P<0.01). Compared with the model group, Brdu positive cell rate increased significantly in edaravone group and DLXNF group, and the expressions of Hes1, Hes5 mRNA and protein increased significantly (P<0.01). The expression of Hes1 mRNA in DLXNF group was superior to that in edaravone group (P<0.01), and other indexes had no significant difference.Conclusion DLXNF can promote the proliferation of NSCs in SVZ in cerebral ischemia-reperfusion injury model rats, and up-regulate the expressions of Hes1 and Hes5, whose mechanism may be related to the activation of Notch signaling pathway.

Objective To study the relationship of proliferation of neural stem cells (NSCs) in SVZ and the expressions of c-jun and c-myc in rats with middle cerebral artery occlusion/reperfusion (MCAO/R) injury model administrated byDanlong XingnaoFormula.Methods The focal cerebral ischemia reperfusion injury models were prepared by longa method. Totally 150 male SD rats were randomly divided into sham-operation group, cerebral model group,Danlong XingnaoFormula low-, medium-, and high-dose groups. The treatment groups were given corresponding dose ofDanlong XingnaoFormula, while the sham-operation group and model group were given the same amount of distilled water 24 h after modeling by gavage, once a day, 7 days in a row. 1 d, 3 d and 7 d after reperfusion, modified Neurological Severity Scores (m-NSS) was used to grade neurologic impairment. 7 d after reperfusion taken to the SVZ brain tissue of ischemia side, Brdu immunohistochemical method was used to record the BrdU positive cells number. The hippocampal c-jun, c-myc mRNA and protein expressions were determined respectively by RT-qPCR method and Western blot method.Results Grades of neurologic impairment in others groups were improved obviously than sham-operation group (P<0.01); 3 d, and 7 d after reperfusion, grades of neurologic impairment inDanlong XingnaoFormula groups were obviously lower compared with model group (P<0.05,P<0.01). Brdu positive cell rates in others groups increased obviously compared with sham-operation group; Compared with model group, Brdu positive cell rates inDanlong XingnaoFormula groups increased obviously (P<0.01). The expressions of c-jun and c-myc protein and mRNA inDanlong XingnaoFormula groups improved obviously than sham-operation group and model group (P<0.01).ConclusionDanlong Xingnao Formula can improve the neural function after cerebral ischemia and stimulate the proliferation of NSCs, and the mechanism may be related to activating the expression of c-jun and c-myc and extending the duration.

Tumor metastasis is one of the important biological characteristics of malignant tumor,which is closely related with the prognosis of the cancer patients.High expression of ADAM8 in varieties of tumors was revealed in many recent studies,and such aberrant expression played a crucial role in regulating of tumor metastasis.Studies showed that overexpression of ADAM8 attenuated the intercellular adhesion effect,promoted tumor angiogenesis,and enhanced the degradation of ECM as well as the releasing of cytokines.Therefore,suppression of ADAM8 may lead to inhibition of tumor metastasis,which makes ADAM8 a particular attractive target as it can be used as a prognostic indicator and a potential therapeutic target of malignant tumor.A review about the relations between ADAM8 protein′s abnormal expression and tumor occurrence was discussed in this paper,also include discussion about the mechanisms of ADAM8 protein′s disorder-induced tumor formation,as well as therapeutic strategies based on ADAM8-targeted,which may provide references for follow-up research and clinical treatment.

The study on tumor metabolism has been gradually be-come a hot spot in recent years .A lot of proteins involved in the regulation of tumor metabolism especially the glucose transporter protein 1(GLUT1).As a key regulatory factor mediating energy metabolism within tumor cells , GLUT1 can regulate the glucose intake and maintain the basic level of metabolism in tumor cells . More importantly, the abnormal expression of GLUT1 was asso-ciated with many kinds of tumors , of which GLUT1 was used to meet the energy requirement for the fast growth of tumor .Thus GLUT1 also played a crucial role in growth , differentiation and metastasis of tumor cells and prognosis of tumors .Meanwhile , as three-dimensional crystal structure of GLUT 1 was determined , it is possible to design the small molecular inhibitors of GLUT 1, which can realize “starve to death” tumor cells.GLUT1 can be a particularly attractive target for tumor treatment and interfer-ence.The relationship between abnormal expression of GLUT 1 protein and tumor metabolism was reviewed . Moreover , the mechanism of tumor metabolism regulated by GLUT 1 protein ex-pression and treatment of cancers were discussed , which may provide references for future research and clinical treatment .

Aim To discuss the impact of important ac-tive ingredient of garlic diallyl sulfide———DATS on platelet activating factor (PAF ) mediated melanoma metastasis and its mechanisms.Methods ①MTT was used to test the effect of different concentrations of DATS on B16F10 and A375 melanoma cell growth number;②Scratch test and transwell were employed to test the effect of different concentrations of DATS on B16F10 and A375 melanoma cell migration;③ West-ern blot was used to test the effect of DATS on expres-sion of MMP-2,ERK,p38 induced by PFA;④Intrave-nous injection of tumor metastasis model was used to check the inhibition of DATS in PAF-mediated melano-ma metastasis.Results B16F10 cells relative growth rate fell to 73.21% and 48.78%,respectively,when DATS concentration reached 50 and 100 μmol·L-1 . DATS inhibited the levels of PAF-induced migration of melanoma cells B16F10 and vertical migration signifi-cantly,and inhibited B16F10 cells migration induced by PAF through inhibiting the expression of MMP-2, paxillin protein,FAK and other proteins.Conclusion DATS can significantly inhibit PAF-induced tumor metastasis, which is related to the inactivation of MAPKs.