Objective To investigate the performances in line bisection(LB)and line cancellation(LC)tasks of patients with unilateral neglect.Methods The unilateral neglect patients,performances in LB and LC tasks before and after rehabilitation treatment were retrospectively analyzed.Results Before treatment,statistical analyses revealed the patients,performances in LC and LB tasks did not differ from each other(P =0.902).After treatment,statistical analyses revealed a significant difference between the two tasks(P =0.007).With LB only,29/30(97%)patients with neglect were detected;with LC only,22/30(73%)patients with neglect were detected.Conclusions As the performances of unilateral neglect patients improved,patients revealed difference between the performances in LB and in LC tasks.One test alone would miss some subjects with neglect.Combinative use of the two tests was more sensitive than either of the 2 tests alone.

Research on novel pullulanase has major significance on the domestic industrialization of pullulanase and the breakdown of foreign monopoly. A thermophilic bacteria LM 18-11 producing thermostable pullulanase was isolated from Lunma hot springs of Yunnan province. It was identified as Anoxybacillus sp. by 16S rDNA phylogenetic analysis. Full-length pullulanase gene was cloned from Anoxybacillus sp. LM18-11. The optimum temperature of the pullulanase was between 55 and 60 degrees C with a half-life as long as 48 h at 60 degrees C; and its optimum pH was between 5.6 and 6.4. V(max) and K(m) of the pullulanase was measured as 750 U/mg and 1.47 mg/mL, which is the highest specific activity reported so far. The pullulanase crystals structure showed a typical alpha-amylase family structure. The N-terminal has a special substrate binding domain. Activity and substrate binding were decreased when the domain was deleted, the V(max) and K(m) were 324 U/mg and 1.95 mg/mL, respectively. The pullulanase was highly heterologous expressed in Bacillus subtilis by P43 promoter. The extracellular enzyme activity was 42 U/mL, which increased more than 40 times compared to the initial strain. This pullulanase has good application prospects.
Anoxybacillus ; classification ; enzymology ; China ; Glycoside Hydrolases ; metabolism ; Hydrogen-Ion Concentration ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Temperature

AIM:To investigate the effect of Astragalus polysaccharides ( APS) on chronic heart failure and its mechanism.METHODS:Male SD rats (n=32) were randomly divided into control group , sham group, model group and APS group (8 rats in each group).The left coronary artery ligation in the rats was conducted to establish myocardial infarc -tion heart failure model.After modeling, the rats in APS group were given APS (3 g· kg-1· d-1) by intragastric adminis-tration for 6 weeks.Left ventricular diastolic diameter (LVD), left ventricular systolic diameter (LVS), left ventricular ejection fraction ( LVEF) and fractional shortening ( FS) were detected by echocardiography .HE staining was used to ob-serve the pathological changes .The concentrations of free fatty acid ( FFA) in the serum and myocardium were observed by the method of acetyl coenzyme A synthetase and acetyl coenzyme A oxidase ( ACS-ACOD ) .The protein levels of total AMP-activated protein kinase (AMPK), phosphorylated AMP-activated protein kinase (p-AMPK), fatty acid translocase (FAT/CD36) and carnitine palmitoyltransferase I (CPT-1) were measured by Western blotting.RESULTS: No signifi-cant difference in each index between sham group and control group was observed .Compared with control group , LVEF and FS in model group was significantly decreased , while LVD and LVS was significantly increased ( P<0.05 ) .The LVEF and FS in APS group were significantly improved compared with model group ( P<0.05 ) , and there was no significant difference between APS group and control group .LVD and LVS in APS group were obviously improved compared with mo-del group (P<0.05), and the difference was significant compared with control group (P<0.05).Compared with control group, focal myocardial necrosis increased , and residual myocardial cells reduced in model group , while those was much better in APS group as compared with model group (P<0.05).The FFA concentrations in the serum and myocardium in model group increased significantly compared with control group ( P<0.05 ) , while those decreased significantly in APS group as compared with model group (P<0.05).The protein levels of p-AMPK, CPT-1, and cell membrane FAT/CD36 in model group decreased significantly compared with control group (P<0.05), and those in APS group increased obvious-ly compared with control group (P<0.05).CONCLUSION:APS improves chronic heart failure by activating the AMPK pathway and promoting myocardial ingestion and utiliation of FFA .

[ ABSTRACT] AIM:To study the protective effect of Astragalus polysaccharides ( APS) on free fatty acid-induced injury in human umbilical vein endothelial cells (HUVECs).METHODS: Cultured HUVECs were divided into control group, APS group [ APS (200 mg/L) treated for 24 h], free fatty acid group [free fatty acid (0.25 mmol/L) treated for 24 h], free fatty acid plus APS group [free fatty acid (0.25 mmol/L) and APS (200 mg/L) treated for 24 h], and com-pound C group [ free fatty acid (0.25 mmol/L) and APS (200 mg/L) and AMPK inhibitor compound C (10 μmol/L) treated for 24 h] .The cell viability was detected by MTT assay.Nitric oxide ( NO) content in the medium was determined by nitrate reductase assay.The protein levels of total adenosine monophosphate-activated protein kinase (AMPK), phos-phorylated adenosine monophosphate-activated protein kinase (p-AMPK), endothelial nitric oxide synthase (eNOS) and phosphorylated endothelial nitric oxide synthase ( p-eNOS) were measured by Western blot.RESULTS: No significant difference of all indexes between APS group and control group was observed.The cell viability in free fatty acid group de-creased significantly compared with control group.The cell viability in free fatty acid plus APS group was significantly im-proved as compared with free fatty acid group.The cell viability in compound C group was almost the same as that in free fatty acid group.The No content and protein levels of p-AMPK and p-eNOS in free fatty acid group decreased obviously as compared with control group, while the NO content and protein levels of p-AMPK and p-eNOS in free fatty acid plus APS group increased obviously compared with free fatty acid group.No significant difference of the p-AMPK and p-eNOS protein levels between free fatty acid plus APS group and free fatty acid group was observed.No significant difference of the AMPK and eNOS protein levels in all groups was found.CONCLUSION:APS attenuates the free fatty acid-induced injury, and its mechanism is related to the AMPK-eNOS signal pathway.

Objective To investigate the effect of astragalus polysaccharides (APS) on the growth and proliferation of hepatocellular carcinoma cells and its effect on adenosine monophosphate activated pro-tein kinase (AMPK) activity.Methods Hepatocellular carcinoma HepG2 cells cultured for 12,24,and 48 hours were treated with 200,300,and 400 mg/L concentration of astragalus polysaccharides.The cell inhibition rate was detected with methyl thiazolyl tetrazolium (MTT),and apoptosis was observed under the fluorescence microscope.Western blot method was used to measure the expression of total AMPK,phosphorylated AMPK (p-AMPK),and phosphorylate mammalian target of rapamycin (p-mTOR) protein expressions.Results Astragalus polysaccharides of each concentration significantly inhibited the proliferation of human hepatoma HepG2 cells (P ＜ 0.01),and the effect of 300 mg/L concentration astragalus polysaccharides was more significant than that of the 200 mg/L concentration (P ＜0.01);while inhibitory effect of 400 and 300 mg/L Astragalus polysaccharides on the proliferation of human hepatoma cell line HepG2 was not significant difference.We found that Astragalus polysaccharides of each concentration could promote the apoptosis of HepG2 cells,and the effect of 300 mg/L Astragalus polysaccharides was more significant.However,astragalus polysaccharide of 400 mg/L concentration could promote the apoptosis no more than the 300 mg/L concentration,which was observed by fluorescent microscope.Western blot results showed that astragalus polysaccharides could increase the expression of p-AMPK (P ＜ 0.05),and inhibit its downstream protein expressions of p-mTOR (P ＜ 0.05).The proliferation effect of astragalus polysaccharides was weakened after accession of AMPK antagonist compound C on hepatocellular carcinoma cells.Conclusions APS can inhibit the growth and proliferation of hepatocarcinoma cells,and its mechanism is related to the AMPK-mTOR pathway.

In order to improve the metabolism yield of cordycepin and adenosine,we studied nitrogen sources ,the levels of nitrogen sources and carbon sources,dynamic development in liquid culture ,as well as the total yield of cordycepin and adenosine in culture system through determining the contents of cordycepin and adenosine by HPLC .The results are as follows: not only animal protein but also some plant protein is very good nitrogen sources; in culture solution the suitable levels of nitrogen sources and carbon sources is respectively 3% and 4%; the content of adenosine in culture solution is very low, while it is quite high in mycelia; it is much higher that the total yield of cordycepin in culture solution than in mycelia; when the culture system is oscillated and cultured after 7~9 days, the total yield of cordycepin and adenosine are both high.

Objective To investigate serum total prostate special antigen(tPSA),complexed prostate special antigen(cPSA)and cPSA/tPSA ratio(C/T) in the differential diagnosis of benign prostate hyperplasia(BPH)and prostate cancer(PCa).Methods A total of 184 serum samples from hospital outpatient and inpatient were collected from May 2009 to December 2015,and which were divided into three groups including other diseases 68 patients,benign prostate hyperplasia(BPH group)80 patients,prostate cancer (PCa group)36 patients.Chemiluminescence immunoassay was used to detect the concentration of tPSA and cPSA,and the C/T ratio were calculated in the study.Results The tPSA,ePSA levels and C/T ratio were significant different from patients with PCa,BPH and other diseases group(P＜0.01).Compared with BPH group(r=0.661),the tPSA and cPSA levels have a higher relativity in PCa group(cPSA=0.708 × tPSA-0.219,r=0.956).While when tPSA level ranges 4.0-10.0 ng/mL),no significant difference of tPSA,cPSA levels and C/T ratio were found in these 3 groups(P＞0.05).Conclusion Combined detection of serum tPSA,cPSA and C/T,making importance in the differential diagnosis of BPH and PCa.

Objective To explore the application of evidence-based medicine in rehabilitation therapy in clinical teaching. Methods 45 interns were divided into 2 groups. The interns during July, 2011 to April, 2012 as the control group (n=23) received the traditional teaching method and the interns during July, 2012 to April, 2013 as the test group (n=22) accepted the evidence-based medicine teaching method in addition. They were assessed with the test scores of theory and operation, and questionnaires. Results The operation scores and the total scores were higher in the test group than in the control group (P<0.05). The satisfaction was better in the test group than in the control group in the items of improving the ability of self-study, taking the initiative, developing clinical thinking, improving the ability of obtaining information, improving the ability of selecting literature, improving the ability of analyzing and summarizing, improving their overall level and degree of satisfaction with this teaching method (P<0.05). Conclusion The evidence-based medicine teaching method can facilitate to improve the practical skills, cultivate the clinical thinking and improve the overall level for students in rehabilitation therapy in clinical practice.

Objective To investigate the relationship between carotid intima media thickness (IMT) and pulse pressure index (PPI) in elderly hypertensive patients.PPI was defined as 24 h mean pulse pressure(PP)/24 h mean SBP.Methods One hundred and three elderly hypertensive patients were categorized by PPI level:group A (PPI

OBJECTIVESTo identify hepatic progenitor cells (HPCs) in patients with severe hepatitis (SH) by detecting their markers and investigate the features of their distribution and location.

METHODSLiver tissues taken from 59 SH patients were tested for the receptor of stem cell factor (c-kit), pi-class glutathione S-transferase (GST-pi), cluster of differentiation 34 (CD34), cytokeratin 19 (CK19), cytokeratin 18 (CK18) and alpha fetoprotein (AFP) by immunohistochemistry (IHC). Meanwhile, 58 patients with acute or chronic hepatitis were also detected to act as controls.

RESULTSHepatic progenitor cells could be seen in SH patients. Most of them existed as ductular cells that had been called "typical ductular proliferation (ADP)" or "typical ductular reaction" in previous research. These ductular cells were mainly located at the portal areas, fibro septa, periportal parenchyma and the border of the pseudolobuli and inflammatory foci. Further, c-kit, GST-pi, CK19 and CK18, but not CD34 and AFP could be detected in these cells. Another kind of HPC was the small hepatocyte-like cell (SHLC), which could express c-kit, GST-pi, and CK18, but not CK19, CD34 and AFP. The semi-quantitative analysis showed that the scope of ADP in SH patients was significantly larger than that in acute and chronic hepatitis patients (chi2= 63.62, P<0.05), and the scope of ADP in subacute severe hepatitis and chronic severe hepatitis patients was also significantly larger than that in acute severe hepatitis patients.

CONCLUSIONIn the course of regeneration of viral hepatitis, different types of pathology have different features. In acute and chronic hepatitis (G1-2), the regeneration is mainly owing to the proliferation of mature hepatocytes, and in chronic hepatitis (G3-4), there is the participation of HPCs, although they are limited. In severe hepatitis, however, since the replicative capacity of normal hepatocytes is impaired or prohibited, liver regenerates and restores mainly by the means of hepatic stem cells activation and proliferation. But the hepatic stem cells don't differentiate into their mature functional compartments directly at all. There are several intermediary or transition populations. In human severe hepatitis, they are mainly ductular cells, and parts of them are small hepatocyte-like cells.

Antigens, CD34 ; analysis ; Cell Division ; Female ; Glutathione S-Transferase pi ; Glutathione Transferase ; analysis ; Hepatitis ; pathology ; Hepatocytes ; pathology ; Humans ; Immunohistochemistry ; Isoenzymes ; analysis ; Keratins ; analysis ; Male ; Proto-Oncogene Proteins c-kit ; analysis ; Stem Cells ; pathology ; alpha-Fetoproteins ; analysis