Objective To obtain highly purified botulinum neurotoxin A light chain(BoNT-ALC) protein in E.coli by genetic engineering and multi-step purifications, and identify its metalloproteases activity.Methods The full-length of BoNT-ALC was cloned from BoNT A by PCR and inserted into plasmid pET-22b.Then pET-22b-ALC was transformed into E.coli BL21( DE3) strains and induced by IPTG.The protein was purified by Ni-NTA sepharose,anion exchange column and gel filtration.The enzymatic activity of the protein was identified by SNAP-25.Results and Conclusion A highly purified and homogeneous protein is obtained, which shows good enzymatic activity.

Objective To analyze the clincopathological features and prognosis of 77 colorectal cancer patients with synchronous liver metastases after resection of the primary tumors.Methods Seventy seven colorec-tal cancer patients with synchronous liver metastases who underwent resection of the primary tumors and patholog-ical confirmed were enrolled in this retrospective study.The clincopathological features and prognosis were ana-lyzed.Kaplan-Meier method was used to analyze survival rate.The differences between survival curves were cal-culated by Log-rank test.Results The analysis showed that sex,tumor differentiation and TNM stage were as-sociated with prognosis(P<0.05).The analyses demonstrated no differences in age,primary tumor site,treat-ment,number of liver metastases and extrahepatic metastasis(P>0.05).There was a significant correlation be-tween CEA,CA19-9 level and gender,tumor differentiation and TNM stage(P<0.001)and the corresponding surgical procedure and TNM stage(P<0.05).Conclusion The analysis show that male,the low differentiation and stageⅢare poor prognosis factors.

To investigate physicians' knowledge about chronic obstructive pulmonary disease (COPD) in tertiary hospitals in northeast China.Physicians from 77 tertiary hospitals in northeast China were surveyed with a questionnaire,which included questions such as risk factors,symptoms,exacerbations,comorbidities and diagnostic criteria of COPD.Besides cigarette smoking,air pollution and pulmonary infections,only 22.5％ (40/178) physicians recognized that the biomass fuels may induce COPD.Totally 59.0％ (105/178) physicians recognized the importance of spirometry to the diagnosis of COPD.Besides dyspnea,cough,sputum production,wheezing and chest tightness,only 23.7％ (42/177)of physicians considered that limitation of activity was an important symptom of COPD.65.5％ (116/177)physicians believed that recurrent lung infections was one of the most important comorbidities of COPD.However,less than 30％ [20.9％ (37/177)-28.8％ (51/177)] physicians were aware of the other important comorbidities.The physicians of tertiary hospitals in northeast China need to be systematically educated on COPD to meet the new guideline.

Objective To investigate the effective connectivity of limbic circuit in patients with major depressive disorder when they recognized dynamic positive face expressions,aiming to discuss the possible mechanism of emotion processing in depressed patients.Methods Eighteen depressive patients and eighteen well-matched healthy control volunteers participated in the experiment.All subjects were asked to recognize the emotion face during the magnetoencephalograph (MEG) scanning.The regions of interested (ROI) brain areas included the orbital frontal cortex (OFC),the anterior cingulated cortex (ACC),the amygdala (AMYG),the hippocampus and the insula.The MEG data were preprocessed by the SPM8 software and further analyzed by the Granger casual model (GCM).The non-parameter permutation was used to compare the value of effective connectivity between the healthy controls and the depressed patients.Results Compared with healthy controls,the effective connectivity from the ACC to the AMYG (P=0.0052),from the OFC to the AMYG(P=0.0046),from the Hippocampus to the ACC (P=0.0016),and from the ACC to the Hippocampus (P=0.0042)was significantly reduced in depressed patients.Conclusion The depressed patients display decreased interaction of the limbic circuit during the happy facial emotion processing,indicating that the depressed patients are unable to deal with the positive stimuli,and to certain extent,explaining the abnormal neuropathophysiological mechanism of positive stimuli in MDD.

Objective To explore crossed cerebellar diaschisis (CCD) in cerebral gliomas with three dimensional arterialspin-labeling (3D ASL) perfusion MRI.Methods The images of 31 patients with cerebral gliomas and 31 normal subjects were retrospectively analyzed.The cerebral blood flow (CBF) were measured with 3D ASL technology,and the asymmetry index (AI) of cerebellar hemispheric CBF value changes were calculated and compared in patients.And the relationship between the AI values of cerebellar hemispheric and the AI values of tumor region in cerebral hemispheric,tumor histological grade and size were investigated.Results Compared with tumor ipsilateral cerebellar hemisphere of supratentorial glioma,the CBF value of contralateral cerebellar hemisphere significantly reduced (t=5.04,P＜0.01),and the AI values of cerebellar hemispheric in patients obviously increased compared to normal subjects (t=4.13,P＜0.01).But there was no significant difference in the AI values of cerebellum between high grade and low grade gliomas (t=1.31,P＞0.05).In cerebral gliomas patients,there was no significant correlation between the AI values of cerebellar hemispheres and the AI values of cerebral hemisphere (tumor:r=-0.28;tumor parenchyma:r=-0.24;tumor plus edema:r=-0.19,all P＞0.05),and tumor size (r=0.18,P＞0.05).Conclusion Cerebral glioma can cause CCD phenomenon,and 3D ASL is able to quantitatively assess the degree of cerebellar hypoperfusion noninvasively.This phenomenon may not be associated with tumor histological grade,size and the AI values of cerebral hemispheric.

Objective To assess the anti-arrhythmic activity and cardioprotective effects of Wenxin Granula, a traditional Chinese formula (consisting of Salviae Miltiorrhizae Radix, Polygonati Rhizoma, Notoginseng Radix et Rhizoma, Nardostachyos Radix et Rhizoma, Angelicae Sinensis Radix, and Succinum), on heart in ischemic-induced myocardial infarction (MI) rats and compare with those of Amiodarone which have been demonstrated in clinic. Methods Rats were randomly divided into Sham-operated (control), Ml + Amiodarone [5 mg/(kg·d)] (MI), and MI + Wenxin Granula [10 mg/(kg·d)] groups and left anterior descending coronary artery was occluded in each group. After left anterior descending for 12 h, standard lead Ⅱ of administration electrocardiogram was recorded in order to analyze the occurrence of arrhythmia. After one month, the size of the infarct area of heart was evaluated by TTC staining method and haemodynamic function was assessed to detect the heart function. Laser scanning confocal microscope and the technique of patch clamp were used to detect the intracellular Ca2+ ([Ca2+]j) and L-type calcium current (ICa-L), respectively. Results Both Wenxin Granula [10 mg/(kg·d)] and Amiodarone [5 mg/(kg·d)] could markedly decrease the incidence of arrhythmia in heart of rats which were subjected to ischemic injury. After one month, Wenxin Granula could significantly decrease mortality to 22.22% and reduce the infarct area (P < 0.05), but Amiodarone did not. The mechanism may involve that Wenxin Granula attenuated [Ca2+]j decreasing in MI rats. Additionally, Wenxin Granula could obviously ameliorate the impaired heart function of MI rats by decreasing the elevated left ventricular end-diastolic pressure and increasing the attenuated maximum change velocity of left ventricular pressure in the isovolumic contraction or relaxation period. On the other hand, electrophysiological experiment results revealed that Wenxin Granula administration one month later also increased the reduced ICa-L density in rat ventricular myocytes in MI rats. The results of LSCM showed that Wenxin Granula could recover the amplitude of [Ca2+]j decreased by heart failure during long term. Conclusion Wenxin Granula could not only inhibit the incidence of arrhythmia but also decrease the mortality, which was accompanied by recovering the amplitude of [Ca2+]j. This protective effect of Wenxin Granula may partially be mediated through changing ICa-L.as well as increasing [Ca2+]j.

Objective To investigate the correlation between arachidonate 15-lipoxygenase (ALOX15)gene polymorphism and its genetic predisposition to coronary heart disease (CHD)in Han population of Shaanxi Province so as to provide the basis for early diagnosis and prophylaxis of CHD.Methods The single nucleotide polymorphisms (SNPs)of ALOX15’s rs916055,rs2619112,and rs2664593 were measured by using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF)method in 105 CHD patients (CHD group)and 75 non-CHD patients (control group)who were matched in age and sex.Results The frequencies of genotypes and alleles of SNPs rs916055A/G in CHD group were significantly different from those in control group (P=0.000 1,P=0.000 1).The frequencies of genotypes and alleles of SNP rs2619112A/G in CHD group did not significantly differ from those in control group (P=0.134 2,P=0.143 8).The frequencies of genotypes of SNP rs2664593C/G in CHD group significantly differed from those in control group (P=0.002 7),but the frequencies of alleles were not significantly different (P=0.537 1).Logistic regression analysis indicated that the A allele of SNP rs916055 was an independent risk factor for CHD.Conclusion SNP rs916055 may be related to CHD and its A allele may be the genetic susceptibility gene for CHD.

In 2011,the International Association for the Study of Lung Cancer(IASLC),American Thoracic Society(ATS)and European Respiratory Society(ERS)simultaneously defined micropapillary pattern(MPP)-based lung adenocarcinoma as an independent pathological type:micropapillary-predominant adenocarcinoma(MPA).Its histological and clinical specificities have attracted broad attention from researchers.Lung adenocarcinoma with a micropapillary pattern(MPPAC)often shows biological behaviors indicative of poor prognosis,including strong invasion,high metastasis rate and easy recurrence after surgical treatment.At present,the clinical diagnosis of MPPAC depends on postoperative pathological examination,but systematic studies on its pathological heterogeneity,molecular biological characteristics,clinical features and prognosis remains scarce.This review aims to focus on the research progress of the pathological and clinical features of MPPAC.

Rhesus monkeys (Macaca mulatta) are human's closest evolutionary relatives next to Chimpanzees, and they are widely used in biomedical researches. Analyses of the rhesus monkey trasnscriptome and the sequence divergence between monkey and human are of importantce to the development of scientific analyses and to the application and interpretation of the results from animal experiments. In this study, we analyzed the genetic and transcriptional information. Four hundred and one clones were randomly selected from a liver tissue cDNA library of rhesus monkey, and the expressed sequence tags (ESTs) were sequenced. We acquired 393 effective ESTs that were assembled into 221 Unigenes with Phrap software. Alignments of the sequences showed that 188 Unigenes matched with known proteins in Swiss_prot database, of which 16 Unigenes matched the known rhesus proteins, and 171 Unigenes had high homology with human proteins. Then the result of BLASTN comparison showed that 26 of another 33 Unigenes matched the known rhesus genes. Finally, the remaining Unigenes were aligned in dbEST and rhesus genome database, and the results suggested 3 Unigenes be newly discovered ESTs of rhesus.
Animals ; Expressed Sequence Tags ; chemistry ; Gene Library ; Liver ; chemistry ; Macaca mulatta ; genetics ; Sequence Analysis, DNA

Erythropoietin-producing hepatocellular A2 receptor (EphA2) is the most common subtype in the largest subfamily of the receptor tyrosine kinase superfamily, and is considered as a key factor in the regulation of malignant tumor progression. EphA2 is highly expressed in glioma, which plays an important role in the development and progression of glioma. This article reviews the structure, function, expression of EphA2 in glioma, and its role in glioma cell migration, maintenance of glioma stem cells, angiogenesis and targeted therapy.