Objective To investigate the localization coexpression in situ of matrix metalloproteinase(MMP)-2,-8 and vascular endothelial growth factor(VEGF)in human atherosclerotic unstable plaques with monocytes,smooth muscle cells(SMCs)and endothelial cells(ECs).Methods The histopathologic changes of unstable human atherosclerotic plaques were observed by hematoxylin and eosin(HE)staining,and the localization coexpression of MMP-2,MMP-8 and VEGF in the unstable human atherosclerotic plaques were observed by double fluorescent immunochemistry technology and confocal microscopy.Results The human atherosclerotic plaques in 6 cases had typical histopathologic instability,which was classified as super-Ⅳ type unstable plaques.The MMP-2 coexpression was the most obvious in the smooth muscle cells of fibrous cap infiltrated by monocyts,and in the monocytes of shoulder of plaques,and more expression of MMP-2 in the microvascular endothelial cells at the edge of shoulder and lipid necrosis;MMP-8 coexpressed obviously with the monocytes in the fibrous cap and lipid cores of plaques,and next to coexpressing in the smooth muscle cells of fibrous cap,while coexpression in endothelial cells was very little;VEGF coexpression was significant in the proliferative microvascular endothelial cells of plaques;The fibrous cap,which consisted of the smooth muscle cells mainly,and the edge of lipid necrosis infiltrated by more monocyts,were over-expression areas of VEGF.Conclnsions MMP-2,-8 and VEGF can coexpress with monocytes,SMCs and ECs in unstable plaques,and the major expression areas are in the fibrous cap,shoulder and micro-vessel at the edge of lipid necrosis,which are infiltrated by monocytes.Moreover,the outer membrane of vessel is involved in the pathogenesis of atherosclerosis.

Objective To investigate the depression in stroke patients with broca aphasia.Methods Aphasia Depression Rating Scale (ADRS) and Zung's self-rating depression scale (SDS) were applied in patients on the first stroke with broca aphasia (Aphasia group,66 cases).The results were compared with stroke patients without aphasia(control group,66 cases).Results Compared to control group,the incidence of depression in Aphasia group was significantly higher(62.12% vs 25.76%,P

Objective To determine pH value, the solubility in different solvents and the oil-water partition coefficients of cumarin. Method UV spectrophotometry was used to determine the content of cumarin. Shake-flask method was developed to determine the oil-water partition coefficients of cumarin. Result The higher solubility values of cumarin were 2 044,2015 and 1969 ?g/mL in acetone , methanol and ethanol, it was almost indiscerptible in water, the limit solubility was 1096.9 ?g/mL, which become stable while the whisk time reached to 8 hours. The oil/water detached coefficient was 11.210, which was higher in acid solvent. Conclusion The method is simple, rapid and reliable.

Atherosclerosis is the most important pathological basis of ischemic cerebrovascular diseases. As an independent risk factor of atherosclerosis, fibrin(ogen) and its degradation products involve in the processes of the formation and development of atherosclerosis. This article reviews the relationship between fibrin(ogen) and/or its degradation products and atherosclerosis, and also introduces the development and application prospects for some specific motif antagonists of fibrin(ogen) in the treatment of atherosclerosis and ischemic cerebrovascular diseases.

Objective To investigate the influence of aging on motor function,binding activity and protein expression level of striatum dopamine transporter(DAT)of rats. Methods Rolling-bar test was performed to assess motor function.Western blot and 131I-FP-β-CIT up-take ratio were used to evaluate DAT protein 1evel and striatum DAT binding activity respectively. Results There were an age-related decline of rolling-bar latency and striatum 131I-FP-β-CIT up-take ratio in rats older than6 months.There was a significant difference between 6-month-old rats and older rats(12-month-old rats,16-month-old rats,20-month-old rats),and rolling-bar latency was correlated with striatum 131I-FP-β-CIT up-take ratio(r=0.656,P＜0.01).There was no change in DAT protein 1evel during aging process. Conclusions The age-related decline of motor function of rats was correlated with the decreasing of striatum DAT binding activity.The synaptic membrane expression of striatum DAT may decrease in aged rats.

Objective To evaluate the value of diffusion weighted imaging (DWI)in distinguishing the solid solitary pulmonary nodule (SPN).Methods 42 patients with SPN (malignant in 25 and benign in 1 7)who were confirmed by operation,biopsy or follow up after treatment underwent routine chest T1 WI,T2 WI and DWI.The b values were chosen as 300,500,800 and 1 000 s/mm2 ,and the corresponding apparent diffusion coefficient (ADC)values and the signal intensity (SI)were respectively measured.Results The ADC values and SI of benign and malignant SPNs were gradually reduced with increasing b value.The ADC value between benign and malignant SPNs was statistically significant with b value of 500 s/mm2 (P 500 =0.03 <0.05 ),meanwhile the SI was statistically significant with b values from 300 to 1000 s/mm2 (P 300 <0.001,P 500 =0.03 <0.05,P 800 =0.01 <0.05, P 1 000 =0.02<0.05).Conclusion Both SI and ADC value of DWI play important role in distinguishing benign and malignant SPNs, and the diagnostic efficiency of SI is superior to ADC value.

Objective To observe the effect of mutant α-synuclein(A30P)in autophagic programmed cell death by transfected PC12 cells and explore its probable role and pathway in PD.Methods The definite PC12 cells which were transfected mutant α-synuclein(A30P)were constructed at first and MPP+,Rapamycin and Wortmanin were administrated to transfected PC12 cells with mutant α-synuclein. Not only the proliferative activity of cells was detected with MTT method but also the ultrastructttre changes of cells and expression of α-synuclein in different circumstance were observed by transmission electron microscopy(TEM),Western Blot and the level of SOD.Results (1)The expression of α-synuclein in groups A30P+Wortmannin and A30P+MPP+was higher than that in group A30P(P＜0.01), particularly.there was more significant expression of α-synuclein in group A30P+Wortmannin.The expression of α-synuclein in group A30P+Rapamycin was weaker than that in group A30P(P＜0.01); (2)The results showed that the SOD level(group A30P+MPP+:3 h:97.49±13.8;12 h:102.7±12.7; 24 h:101.5±11.8;48 h:104.3±12.4)was significantly decreased at various time points after MPP+ treatment compared that of group A30P(t=3.7721,P=0.0017).SOD level gradually increased in A30P +Rapamycin 12 h and showed significant difference at 24 h(121.2±13.0),48 h(124.3±14.1)and 72 h(127.7±13.7)after drug treatment compared with that in group A30P+Wortmannin(t:2.9746, P=0.0083);(3)Mutant α-synuclein(A30P)leading to PC12 cells death by means of autophagy involved α-synuclein accumulation,membrane lipid oxidation,and loss of plasma membrane integrity.Mutant α- synuclein(A30P)mediated the toxicity of MPP+.Rapamycin,an inducer of autophagy,reduced the aggregation of α-synuclein in transfected cells.Meanwhile,Wortmanin,an inhibitor of autophagy,promoted the aggregation of α-synuclein in transfected cells and induced cells to die.Conclusions The abnormal aggregation of α-synuclein induces autophagic programmed cell death in PC12 cells and mutant α-synuclein (A30P)mediates the toxicity of MPP+.Meanwhile,Rapamycin may reduce the aggregation of α-synuclein in transfeeted cells by activation of autophagic pathway.

Objective:To explore the key amino acids of side chain dioxate dehydrogenase complex E2 protein (BCOADC-E2) that can react specifically with specific autoantibody anti-mitochondrial antibodies (AMA)-M2 in patients with primary biliary cholangitis (PBC).Methods:The homologous target gene BCKD, which expressed the epitopes of BCOADC-E2 protein, was cloned and recombined with the engineering plasmid pGEX-4T1. Fifteen point mutant plasmids were obtained by polymerase chain reaction (PCR) and transferred to the prokaryotic expression strain for protein expression and purification. Fourteen mutant proteins and one wild-type protein were obtained. The AMA-M2 specificity of the 14 mutant proteins was measured by enzyme-linked immuno sorbent assay (ELISA), and the amino acids that were critical to the specificity of BCOADC-E2 and AMA-M2 were identified by comparing the specificity of the 14 mutant proteins with that of the wild type proteins. Differences between groups were analyzed by analysis of variance, LSD- t test. Results:A total of 14 mutant proteins and 1 wild-type protein were obtained.The specific reaction degree of mutant protein pGEX-BCKD-S1A (1.634±0.328) and pGEX-BCKD-C3A (1.744±0.345) with serum AMA-M2 in patients with PBC was higher than that of wild-type protein pGEX-BCKD (1.000±0.000) with AMA-M2; Mutant protein pGEX-BCKD-E4A (0.157±0.067), pGEX-BCKD-V5A (0.057±0.029), pGEX-BCKD-Q6A (0.580±0.166), pGEX-BCKD-S7A (0.744 ±0.125), pGEX-BCKD-D8A (0.351 ±0.135), pGEX-BCKD-S10A (0.496 ±0.158), pGEX-BCKD-V11A(0.149±0.089), pGEX-BCKD-T12A(0.061±0.043), pGEX-BCKD-I13A(0.007±0.017), pGEX-BCKD-T14A (0.198±0.101), pGEX-BCKD-S15A (0.156±0.087), The specific reaction degree of pGEX-BCKD-R16A (0.884±0.099) with AMA-M2 was lower than that of wild-type protein pGEX-BCKD (1.000±0.000) with AMA-M2.Conclusion:The cysteines at positions 1 and 3 of BCOADC-E2 protein were the key amino acids to improve the specific reaction between BCOADC-E2 and AMA-M2. The mutant proteins formed after amino acids at position 4, 5, 6, 7, 8, 10, 11, 12, 13, 14, 15 and 16 are replaced by alanine can decrease the specificity of AMA-M2. Amino acids at positions 5 and 13 are the key amino acids that affect the specific reaction between BCOADC-E2 and AMA-M2, and have an important effect on the function of BCOADC-E2 protein.

Objective:To evaluate the effect of methane on acetaminophen-induced acute liver injury (ALI) in mice and the role of autophagy.Methods:Forty clean-grade SPF healthy adult male C57BL/6 mice, aged 8-10 weeks, weighing 20-25 g, were divided into 4 groups ( n=10 each) using a random number table method: control group (group C), group ALI, methane-rich saline group (group MS) and methane-rich saline plus 3-methyladenine (3-MA) group (group MS+ 3-MA). Acetaminophen 300 mg/kg was injected intraperitoneally to establish ALI model.Group MS was injected intraperitoneally with methane-rich saline 10 ml/kg immediately after establishing the model and at 12 h after establishing the model.Group MS+ 3-MA was injected intraperitoneally with methane-rich saline 10 ml/kg and autophagy inhibitor 3-MA 30 mg/kg immediately after establishing the model and was injected intraperitoneally with methane-rich saline 10 ml/kg at 12 h after establishing the model.The equal volume of sterile saline was given intraperitoneally at the same time points in C and ALI groups.At 24 h after establishment of the model, blood samples from the eyeball were taken for measuring concentrations of alanine transaminase (ALT) and aspartate transaminase (AST) in serum (using biochemistry analyzer) and the concentrations of tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serum (using enzyme-linked immunosorbent assay). The animals were then sacrificed and liver tissues were removed for determination of microtubule-associated protein 1 light chain 3 (LC3) and p62 (by Western blot) and for the examination of the number of autophagosomes (under a transmission electron microscope). Results:Compared with group C, the concentrations of ALT, AST, TNF-α and IL-6 in serum were significantly increased, LC3Ⅱ/LC3Ⅰ ratio was increased, expression of p62 was up-regulated, and the number of autophagosomes was increased in liver tissues in ALI, MS and MS+ 3-MA groups ( P<0.05). Compared with group ALI, the concentrations of ALT, AST, TNF-α and IL-6 in serum were significantly decreased, LC3Ⅱ/LC3Ⅰ ratio was increased, expression of p62 was down-regulated, and the number of autophagosomes in liver tissues was increased in group MS, and AST concentration in serum was decreased and LC3Ⅱ/LC3Ⅰ ratio was increased in group MS+ 3-MA ( P<0.05). Compared with group MS, the concentrations of ALT, AST, TNF-α and IL-6 were significantly increased, the LC3 II/LC3 I ratio and the number of autophagosomes were decreased in lung tissues in group MS+ 3-MA ( P<0.05). Conclusion:The mechanism by which methane can reduce acetaminophen-induced ALI is related to enhancement of the level of autophagy in liver cells in mice.

Objective To explore the value of serum IL-11 in the diagnosis and prognosis evaluation of acute cerebral infarction and its correlation with serum brain-derived neurotrophic factor(BDNF).Methods General clinical data of 102 patients with acute cerebral infarction(cerebral infarction group)and 64 normal controls(normal control group)were collected.According to the 90 d mRS score,the cerebral infarction group was divided into the good prognosis subgroup and the poor prognosis subgroup.Pearson correlation analysis was used to analyze the correlation between serum IL-11 and NIHSS score,cerebral infarction volume and serum BDNF.Logistics regression analysis was used to analyze the influencing factors of cerebral infarction prognosis,and the ROC curve of IL-11 in the diagnosis and prognosis of cerebral infarction was drawn.Results The rate of hypertension and the levels of glycosylated hemoglobin and low-density lipoprotein in the cerebral infarction group were significantly higher than those in the normal control group(all P＜0.05).The age,rate of diabetes,glycosylated hemoglobin level,NIHSS score at admission and cerebral infarction volume in the poor prognosis subgroup were significantly higher than those in the good prognosis subgroup(all P＜0.05).The level of serum IL-11 in the cerebral infarction group was significantly lower than that in the normal control group(t =10.123,P＜0.05).The serum IL-11 level in the poor prognosis subgroup of the cerebral infarction group was significantly lower than that in the good prognosis subgroup(t =7.438,P＜0.05).The expression of serum IL-11 in patients with cerebral infarction was negatively correlated with NIHSS score(r =-0.603,P＜0.001)and cerebral infarction volume(r =-0.681,P＜0.001).Logistics regression analysis showed that IL-11 was a protective factor(OR =0.814,P =0.009),while infarct volume(OR = 2.262,P＜0.001)and NIHSS score(OR =2.107,P =0.006)were risk factors affecting the prognosis of patients with cerebral infarction.When IL-11 was applied to the diagnosis of cerebral infarction,the area under the ROC curve was 0.841,the sensitivity was 91.18%,the specificity was 72.42%,and the cut-off value was 378.47;when IL-11 was applied to the prognostic discrimination of cerebral infarction,the area under the ROC curve was 0.786,the sensitivity was 67.09%,the specificity was 87.93%,and the cut-off value was 310.94.The correlation coefficient between serum IL-11 levels and serum BDNF levels in patients with cerebral infarction was r =0.711,P＜0.001.Conclusions The level of serum IL-11 in patients with cerebral infarction is significantly decreased,and the level of IL-11 in patients with poor prognosis is significantly lower than that in patients with good prognosis.Meanwhile,the level of IL-11 is negatively correlated with the level of serum BDNF,which may be used for the auxiliary diagnosis and prognosis evaluation of cerebral infarction.