Objective To explore the neural damage induced by acute exposure to methamphetamine (METH).Methods The mice were administrated with METH,then the stereotyped behavior of mice was evaluated,and spatial recognition memory was analyzed by Y-maze test.In addition,nitric oxide synthase (NOS) activity was detected by kit,and the apoptotic proteins including Bax,Bcl-2,Caspase-3 were assayed by using Western blot.The DNA injury induced by METH was observed by using the comet assay.Moreover,mitochondrial membrane potential was detected to assess the toxic effects of METH on mitochondria by JC-1.With the Western blot assay,the phosphorylation of MAPK signaling pathways were also investigated.Results Acute METH exposure significantly increased the stereotyped behavior in mice,and spatial recognition ability of mice was obviously decreased.On the molecular level,total nitric oxide synthase (TNOS) and induced nitric oxide synthase (iNOS) were increased,and the apoptotic proteins,such as Bax and cleaved caspase-3 were markedly enhanced.With the comet assay,it showed that METH exposure resulted in DNA damage.In parallel,mitochondrial membrane was damaged which manifested as mitochondrial membrane potential decreased.With the western blot,It was further found that METH enhanced the activation of MAPKs.However,p38 MAPK signahng pathway was demonstrated to be the only one factor involved in METH-induced neural damage.Conclusion METH induced neural damage,and MAPK signaling pathways might be involved in this process,since inhibition of p38 MAPK signaling pathway significantly ameliorated METH-induced neural damage.

Objective To investigate the changes in high voltage-activated (HVA) calcium current in dorsal root ganglion (DRG) neurons isolated from rats with neuropathic pain. Methods Pathogen-free male SD rats aged 4-6 weeks weighing 180-220 g were used in this study. The animals were anesthetized with intraperitoneal pentobarbital sodium 50 mg/kg. Neuropathic pain was induced by ligation of L5 spinal nerve between DRG and sciatic nerve. The nerve was transected distal to the ligature. The animals which showed positive signs of neuropathic pain were decapitated on the 14th postoperative day. L5 and L4 DRGs were isolated and the neurons in the ganglia were enzymatically dissociated (group L5 and L4). The control group received no surgery (group C). The HVA Ca2+ current was recorded using whole-cell patch clamp technique. Results Peak calcium current density was significantly lower in group L5 and L4 than in group C, and was significantly lower in group L5 than in group L4 . Halfactivation value (Va 1/2) was also significantly lower in group L5 than in group C and L4 (P ＜ 0.05). The relative contribution of N-type to the total HVA Ca2+ current was significantly greater in group L5 than in group C and L4(P ＜ 0.05). There was no significant difference in the steady-state inactivation curves among the 3 groups. Conclusion In rats with neuropathic pain, the HVA Ca2+ current in the injured DRG neurons may play a key role in the induction of neuropathic pain.

The stems and branches of Hypericum petiolulatum were extracted by alcohol and liquid-liquid extraction. Seven furofuran lignans were isolated from the ethyl acetate fraction of ethanol extract of H. petiolulatum by using silica gelchromatography, Sephadex LH-20 chromatography, medium-pressure liquid chromatography and preparative HPLC. Their structures were identified by the spectroscopic methods as pinoresinol (1), medioresinol (2), 8-acetoxypinoresinol (3), epipinoresinol (4), (+)-syringaresinol (5), (+)-1-hydroxysyringaresinol (6) and erythro-buddlenolE (7). All the isolates were firstly found in H. petiolulatum. In the bioassay, compound 7 showed remarkable antioxidative activity inhibiting Fe(+2)-cystine induced rat liver microsomal lipid peroxidation with inhibitory rate 38% at a concentration of 1 x 10(-6) mol · L(-1) (positive control Vit E with the inhibitory rate of 35% at the same concentration).
Animals ; Antioxidants ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Hypericum ; chemistry ; Lipid Peroxidation ; drug effects ; Microsomes, Liver ; drug effects ; metabolism ; Molecular Structure ; Oxidative Stress ; drug effects ; Plant Stems ; chemistry ; Rats

Objective To investigate the relationship of albumin levels with the prognosis and severity of illness in elderly sep-sis patients.Methods This was aretrospective study.108 elderly sepsis patients were enrolled from October 2014 to Decem-ber 2015.All patients were divided into survivors group (83 cases)and death group (25 cases)based on the 28-day progno-sis.The differences of clinical data and laboratory were compared between two groups.According to the albumin levels,all patients were divided into three groups,normal albumin group (≥35 g/L,24 cases),and mild hypoproteinemia group (28 g/L≤ALB<35 g/L,52 cases)and severe hypoproteinemia group (<28 g/L,32 cases),respectively.The mortality rate was compared in the groups.Spearman Correlation Coefficient was used to analyze ALB and other factors.Results In all 108 pa-tients,the 28-day mortality rate was 23.1%,and 77.8% of the patients with hypoproteinemia.ALB level [31.7 (28.3~35.7)g/L vs 25.8 (21.7~31.8)g/L,P<0.001]and Hb [128 (110~140)g/L vs 102 (84~132)g/L,P=0.015]in death group were significant lower than that in survival group.Meanwhile,APACHE II scores [20 (18~23)vs 22 (19~24),P=0.015]and SOFA score [6 (5~6)vs 6 (6~7),P<0.001]were higher than that in survival group.The mortality decreased with the increase of ALB level (43.8% vs 21.2% vs 0%,P<0.05).ALB had a negative correlation with APACHEⅡ score (r=-0.190,P=0.049)and coma (r=-0.311,P=0.001),and had a positive correlation with Hb (r=0.449,P<0.001).Conclusion The incidence of hypoproteinemia was high in elderly sepsis patients.ALB level was associated with prognosis and severity of illness among the patients.

Objective To investigate the effect of gabapentin on high voltage active calcium currents in the injured dorsal root ganglion (DRG) neurons in a rat model of neuropathic pain.Methods Pathogen-free male SD rats aged 4-6 weeks were used in this study. The animals were anesthetized with intraperitoneal pentobarbital soclium 50 mg/kg. L_5 spinal nerve was ligated between DRG and sciatic nerve and cut distal to the ligature. The animals were decapitated on the 14th postoperative day. L_5 DRG was isolated and the neurons in the ganglion were enzymatically dissociated. The high voltage active calcium current was recorded using whole-cell patch-clamp technique.Results Gabapentin inhibited the peak calcium current in the injured DEG neurons. Peak calcium current was decreased by gabapentin 100 μmol/L and both activation and steady-state inactivation curve shifted to more hyperpolarized potentials. Conclusion Gabapentin can inhibit high voltage active in the injured DRG neurons in a rat model of neuropathic pain. The alteration in the inactivation of the electrophysiological properties may be involved in the mechanism.

BACKGROUND: 5-lipoxygenase protein (ALOX5AP) has been recognized as a susceptibility gene for stroke and coronary artery diseases. The present study was to explore the role of this gene in the eastern Chinese patients with ischemic stroke.METHODS: Using a case-control design, we studied 658 patients with ischemic stroke and 704 unrelated population-based controls who were age- and sex-matched. The 658 patients were classified by the Trial of Org 10172 in Acute Stroke Treatment (TOAST). Two single-nucleotide polymorphisms (SNPs) covering ALOX5AP were genotyped.RESULTS: The genotype frequencies of TG of the SNPs rs17222919 located in the promoter of the ALOX5AP gene were significantly higher in patients with ischemic stroke than in controls (OR*=1.34, 95％CI*=1.02-1.75), especially in patients with ischemic stroke caused by small-artery occlusion (SAO) (OR*=1.40, 95％CI*=1.02-1.93). Meanwhile, the genotype frequencies of TG and TG/GG were higher in female patients than in the controls. After specification, the genotype frequencies of TG and TG/GG were higher in the patients than in controls with hypertension. The genotype frequencies of AG and AG/GG of the SNPs rs9579646 located in the intron of the ALOX5AP gene were higher in the controls than in the patients. After specification, the genotype frequencies of TG were higher in the controls than patients without hypertension.CONCLUSION: The present study suggests that sequence variants in the ALOX5AP gene are significantly associated with ischemic stroke.

BACKGROUND: Genetic variations of the 5-lipoxygenase activating protein and leukotriene A4 hydrolase genes that confer an increased risk of ischemic stroke have implicated the family of leukotrienes as potential mediators of ischemic stroke. This study aimed to explore the association of ALOX5,LTA4H andLTC4S gene polymorphisms with ischemic stroke risk in a cohort of Chinese in east China.METHODS: This case-control study consisted of 690 patients with ischemic stroke and 690 controls. Polymorphisms ofALOX5 rs2029253 A/G,LTA4H rs6538697 T/C, andLTC4S rs730012 A/C were genotyped by the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis. The multivariate logistic regression model was used to exclude the effects of conventional risk factors on ischemic stroke.RESULTS: Carriers of C allele in rs730012 were more susceptible to ischemic stroke (OR: 1.37; 95%CI: 1.08-1.73;P=0.009). The rs2029253 GG genotype showed a risk-reducing effect on ischemic stroke (OR: 0.72; 95%CI: 0.55-0.93;P=0.013) while the rs6538697 CC genotype had an increased risk of ischemic stroke (OR: 1.77; 95%CI: 1.09-2.89;P=0.022). The rs730012 variant was not associated with ischemic stroke risk after adjusting confounding factors (P>0.05).CONCLUSION: The present study suggested that gene polymorphisms in the leukotrienes pathway may exert infl uences, with independent genetic effects, on ischemic stroke susceptibility in a cohort of Chinese in east China.

Objective: To explore the determinants for health-seeking behavior of the residents after cough for more than 3 weeks in Yiwu, Zhejiang Province, in order to provide reference for prevention and control of respiratory diseases. Methods: A multi-stage cluster random sampling method was used to recruit the community residents aged 5 years and above in Yiwu. A face-to-face questionnaire survey was conducted to collect demographic information, features of cough and health-seeking behaviors in the past month. The multivariate logistic regression model was employed to analyze the associated factors for health-seeking behavior after a cough for more than 3 weeks. Results: Among 6 374 residents investigated, 152 cases had a cough for more than 3 weeks in the past month, accounting for 2.48%. They were( 45.00±21.15 ) years old, including 70 ( 46.05% ) males and 82 ( 53.95% ) females. About 58.55% ( 89 ) of them sought medical treatment. The results of multivariate logistic regression analysis showed that females ( OR=2.100, 95%CI: 1.005-4.391 ), middle school education level ( OR=0.406, 95%CI: 0.168-0.983 ), family annual income of 100 000 to 199 999 yuan ( OR=2.993, 95%CI: 1.215-7.373 ) were associated factors for health-seeking behavior after a cough for more than 3 weeks. Conclusion The rate of health-seeking behavior after a cough for more than 3 weeks among the residents in Yiwu is 58.55%, which is associated with gender, education level and income.

The glycine amino peptidase of Actinomucor elegans was studied in this work. For the enzyme production Actinomucor elegans was cultured with an enzyme producing medium. Then the cells were collected and subjected to enzyme purification. The glycine aminopeptidase was purified 592 times by a DEAE-Toyopearl column, a Toyopearl HW 65-C column and a Superdex 200 column subsequently and the purified enzyme had a specific activity of 14.2 u/mg. The enzyme was estimated to have molecular mass of 320kD by gel filtration and a subunit size of 56.5kD by SDS-PAGE. It hydrolyzes glycine residue containing substrates such as glycine-betanaphthylamine more efficiently than those containing other amino acid residue. Addition to Gly-betaNA, the enzyme could also hydrolyze Ala-betaNA, Met-betaNA, Leu-betaNA, Arg-betaNA and Ser-betaNA but it had no activity on the substrates such as Trp-betaNA, Pyr-betaNA, Pro-betaNA, Asp-betaNA, Lys-betaNA, Val-betaNA. It was also observed when the glycine-betanaphthylamine concentration was higher than 2mmol/L the enzyme showed a substrate inhibition, and at the 20 mmol/L the enzyme only showed about 55% activity as it showed at the 2mmol/L. Whereas no such phenomenon was observed on the other substrate such as alanine-betanaphthylamine. The optimal temperature and pH for the reaction of this enzyme is 30 degrees C and pH 8.0, respectively. The Km and Kcat of the enzyme for glycine-betanaphthylamine is 0.24 mmol/L and 100.8 s(-1), respectively. Zn2+, Cu2+ and Cd2+ suppress almost all activities of the enzyme at the concentration of 1.0 mmol/L. Based on the study of chelating reagents, GAP belongs to the metalloenzyme. When a gelatin solution was hydrolyzed with 0.5% of alkaline proteinase together with glycine aminopeptidase at 50 degrees C for 18 hours, the glycine aminopeptidase could improve the hydrolysis degree of the protease. The total free amino acid was improved about 13% and although the enzyme mainly had the activity to hydrolyze the glycine residue, individual amino acids analysis with an amino acid analyzer showed that the contents of glycine, proline, alanine, arginine and glutamate were considerably increased. The results of this study showed that the glycine aminopeptidase would be useful in the food industry.
Aminopeptidases ; antagonists & inhibitors ; isolation & purification ; metabolism ; Catalysis ; Hydrogen-Ion Concentration ; Molecular Weight ; Mucorales ; enzymology ; Temperature

BACKGROUNDLeukotrienes are arachidonic acid derivatives long known for their inflammatory properties. Leukotriene-based inflammation has been demonstrated to play a crucial role in atherosclerosis, a major risk factor for several human diseases. Recently, human genetic studies from us and others suggest that single nucleotide polymorphisms (SNPs) in leukotriene pathway genes influence the risk of atherosclerotic diseases such as stroke. This study aimed to assess the role of additional leukotriene pathway genes as a stroke risk factor within the Chinese Han population.

METHODSWe sequenced the promoter, exonic, and intronic regions of leukotriene A4 hydrolase (LTA4H) and arachidonate 5-lipoxygenase (ALOX5), and then genotyped five SNPs in LTA4H and four SNPs in ALOX5 among 691 cases with stroke and 732 controls from the Chinese population.

RESULTSWe detected a significant association between an intronic SNP in LTA4H (rs6538697) and stroke in our subjects (adjusted odds ratio, recessive model, 1.75; P = 0.022); and the SNP rs2029253 in ALOX5 was associated with a decreased risk of stroke (adjusted odds ratio, 0.76; 95% confidence interval, 0.59 - 0.97).

CONCLUSIONGenetic variants in LTA4H and ALOX5 may modulate the risk of stroke in the Chinese Han population.

Aged ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Female ; Humans ; Leukotrienes ; biosynthesis ; Logistic Models ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Risk ; Stroke ; etiology ; genetics