Halomonas sp.BYS-1 was a moderately halophilic bacteria strain isolated from activated sludge,It could grow on MM with NaCl concentration from 0.1～2.6 mol/L and phenyl acetic acid as sole carbon souce. When BYS-1 grew in the media with different concentrations of NaCl, there was no obvious change in its intracellular Na+ contents , it accumulated K+, glutamic acid and betaine as osmoprotectants. Its intracellular contents of K+,glutamic acid and betaine increased by 1.9,2.4 and 13.6 times, respectively, when the concentration of NaCl increased from 0.1 mol/L to 2mol/L.

Objective To investigate the effect and safety of duct-to-mucosa anastomosis technique for pancreaticoduodenectomy(PD).Methods The clinical data,including pancreatic fistula and other complications,of 189 patients underwent PD with end-to-side pancreaticojejunostomy at our institution from Jan 2001 to Jan 2009 were analyzed retrospectively.The definition of pancreatic fistula was threefold increase over the serum amylase level 7 days after operation,and draining volume was more than 50 ml per day.Results Totally 177 Whipple procedures were performed,while 12 pylorus-preserving pancreateduodenectomy procedures were performed.Five patients developed pancreatic fistula with a incidence of 2.65%(5/189).In which 3 were mild Cases,who fully recovered after conservative management,and the other 2 cages were cured by surgical intervention.Other complications included 9 cases of wound infection(4.76%,9/189),11 cases of empty dysfunction(5.82%,11/189),5 Cases of delayed hemorrhage(2.65%,5/189),and 4 cases of intra-abdominal infection(2.12%,4/189),and 2 patients died due to severe intra-abdominal infection and acute pulmonary infarction.Conclusions Duet-to-mucesa anastomosis technique resembles physiological state with low incidence of pancreatic fistula and delayed anastomosis hemorrhage.It may be used for different kinds of anastomosis for pancreatic stump.

Objective To identify the diffusion alterations of deep gray matter(GM) and white matter (WM) among Alzheimer's disease (AD), mild cognitive impairment (MCI) and healthy people by atlas?based analysis (ABA), and to investigate the respective relationship with cognitive function. Methods Twenty?one AD patients (AD group), 8 MCI patients (MCI group) and 15 normal controls (control group) were performed by conventional MRI and diffusion tensor imaging (DTI). The raw data of DTI was processed by using DTI studio software to generate the fractional anisotropy (FA) images. Then ABA was used to quantify the FA value in 58 deep GM and WM structures. The differences of FA value among three groups were compared by using one way ANOVA, with a post?hoc analysis. In AD and MCI groups, the partial correlation was further investigated between mini?mental state examination (MMSE) score and FA value in the brain regions that have significant differences between AD and MCI group or between MCI and control group. Results Compared with control group, AD patients showed wide?spread FA decrease in most deep GM and WM regions (corrected P<0.05). The FA values of the hypothalamus, the fornix, the superior longitudinal fasciculus (SLF) and the cingulum in AD group were significantly lower than those in MCI group (corrected P<0.05). The FA value of the right splenium of corpus callosum (SCC) in MCI group was significantly lower than that in control group (MCI:0.550±0.018 vs. Control:0.585±0.026, P<0.05). In AD and MCI group, the FA values of the left hypothalamus, the right hypothalamus, the left cingulum, the right cingulum, and the left SLF were positively correlated with MMSE scores(r=0.502, 0.515, 0.535, 0.527, 0.512; P<0.05). No significant correlation was found between the FA value of the right SCC, the right SLF, the right fornix/stria terminalis, the right fornix and MMSE scores(P>0.05). Conclusion Based on ABA, this study found the diffusion changes not only in the WM but also the deep GM in AD patients, but only WM diffusion disruptions in MCI group. The decreased FA value in the right SCC appeared early, but had no correlation with the cognitive impairment. The FA value in the hypothalamus, the fornix, the SLF and the cingulum decreased with the disease progression, and correlated positively with the cognition decline.

Objective To investigate the relationship between the antiproliferation effects of aloe-emodin on growth of gastric cancer cells and cell cycle arrest.Methods Human gastric cancer SGC-7901 cells were treated with 2.5,5,10,20,and 40 ?mol/L aloe-emodin for 1—5 d.The cell growth was determined by MTT assay.Cell proliferation and cycle distributions were analyzed by flow cytometry.Western blotting assay was used to detect the changes of cell cycle regulators,cyclins,and cyclin-dependent kinases(CDK).Results Aloe-emodin inhibited the growth of gastric cancer cells in a dose-dependent manner.Treatment of aloe-emodin resulted in cell cycle arresting at G2/M phase.Its molecular mechanisms involved the decrease of the expression of cyclin A and CDK2,the increase of the expression of cyclin B1 and CDK1.Conclusion One of the antitumor mechanism of aloe-emodin on the growth of gastric cancer SGC-7901 cells is to arrest the cell cycle,which indicates that aloe-emodin has a potential value for the treatment of gastric cancer in clinic.

HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular ; Escherichia coli ; metabolism ; HIV Integrase ; metabolism ; HIV-1 ; physiology ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Protein Binding ; Virus Replication

The 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) catalyzes the conversion of HMG-CoA to mevalonate in mavalonic acid pathway, which is the first committed step for isoprenoid biosynthesis in plants. However, it still remains unclear whether HGMR gene plays a role in the isoprenoid biosynthesis in Dendrobium officinale, an endangered epiphytic orchid species. In the present study, a HMGR encoding gene, designed as DoHMGR1 (GenBank accession JX272632), was identified from D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods, for the first time. The full length cDNA of DoHMGR1 was 2 071 bp in length and encoded a 562-aa protein with a molecular weight of 59.73 kD and an isoelectric point (pI) of 6.18. The deduced DoHMGR1 protein, like other HMGR proteins, constituted four conserved domains (63-561, 147-551, 268-383 and 124-541) and two transmembrane motifs (42-64 and 85-107). Multiple sequence alignment and phylogenetic analyses demonstrated that DoHMGR1 had high identity (67%-89%) to a number of HMGR genes from various plants and was closely related to Vanda hybrid cultivar, rice and maize monocots. Real time quantitative PCR (qPCR) analysis revealed that DoHMGR1 was expressed in the three included organs. The transcripts were the most abundant in the roots with 2.13 fold over that in the leaves, followed by that in the stems with 1.98 fold. Molecular characterization of DoHMGR1 will be useful for further functional elucidation of the gene involving in isoprenoid biosynthesis pathway in D. officinale.
Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Dendrobium ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Hydroxymethylglutaryl CoA Reductases ; genetics ; metabolism ; Molecular Weight ; Phylogeny ; Plant Leaves ; enzymology ; genetics ; Plant Roots ; enzymology ; genetics ; Plant Stems ; enzymology ; genetics ; Plants, Medicinal ; enzymology ; genetics ; Sequence Alignment ; Sequence Homology, Amino Acid

Objective To investigate the vascular effect of hydroxyl-safflor yellow A(HSYA) on rat thoracic aorta and its underlying mechanism. Methods The tension of isolated thoracic aorta rings of rats perfused with different concentrations of HSYA(1?10-6-1?10-4 mol/L) was measured using organ bath technique.The effects of HSYA on the vasocontraction induced by cumulative phenylephrine(PE)(1?10-6-1?10-4 mol/L),KCl(6?10-2 mol/L) and CaCl2(1?10-5-3?10-3 mol/L) were recorded respectively. Results HSYAcaused a concentration-dependent anti-contraction effects by KCl or PE in endothelium-intact and endothelium-denuded aortic rings.HSYA inhibited the CaCl2-induced contraction and downward shifted concentration-response curve of aortic rings.HSYA+HP resulted in more significant anti-contraction effect than single use of HSYA(P0.05).There were significant differences in anti-contraction effect between HSYA+RR and RR or HSYA(P

Mitogen-activated protein kinases (MAPKs) are important signaling transduction components well conserved in eukaryotes and play essential roles in various physiological, developmental and hormonal responses in plant. In the present study, a MAPK gene, designated as DoMPK4 (GenBank accession No. JX297597), is identified from a rare endangered medicinal orchid species D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The full length cDNA of DoMPK4 is 1 518 bp in length and encoded a 369 aa protein with a molecular weight of 42.42 kD and an isoelectric point of 5.55. DoMPK4 protein contained a serine/threonine protein kinase active site (158-170), a MAP kinase site (71-174), and eight conserved motifs. DoMPK4 had a transmembrane (214-232) but no signal peptide. Multiple sequence alignment showed that DoMPK4 shared high identities (74.9%-80.6%) with MAPK proteins from various plants. Phylogenetic analysis demonstrated that DoMPK4 belonged to group A of the MAPK evolutionary tree, and is closely related to monocots. Real time quantitative PCR (qPCR) analysis revealed that DoMPK4 is differentially expressed among the five organs including leaf, stem, root, seed, and protocorm-like body (PLB). The transcription level of DoMPK4 is the highest in the PLBs with 17.65 fold, followed by seeds, roots, and stems with 5.84, 2.28, and 1.64 fold, respectively. The progressive enhancement of DoMPK4 transcripts in the developing PLBs compared to that in the germinating seeds, suggests a role of DoMPK4 during the development of embryogenic PLBs formation in D. officinale.
Amino Acid Sequence ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Dendrobium ; enzymology ; genetics ; Gene Expression Regulation, Plant ; Mitogen-Activated Protein Kinases ; genetics ; metabolism ; Phylogeny ; Plant Leaves ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; metabolism ; Plant Stems ; metabolism ; Seeds ; metabolism ; Sequence Alignment

Objective To investigate the prognosis of patients with primary liver cancer in different pathological types after hepatectomy,and to analyze the effects of clinicopathological factors on the survival.Methods The clinical data of 567 patients with primary liver cancer who received hepatectomy at the Affiliated Hospital of Qingdao University from January 1997 to December 2008 were retrospectively analyzed.All patients were divided into hepatocellular carcinoma (HCC) group,cholangiocarcinoma (CC) group and combined hepatocellular carcinoma and cholangiocarcinoma (cHCC-CC) group.The survival and risk factors of the patients were analyzed.All data were analyzed by using the chi-square test,t test,analysis of variance.The survival curve was drawn by the Kaplan-Meier method and the survival of the 3 groups was compared by the Log-rank test.The risk factors were analyzed by the one-way analysis of variance and COX regression model.Results The results of pathological examination confirmed that 92.9％ (527/567) patients were with HCC,4.6％ (26/576) with CC and 2.5％ (14/567)with cHCC-CC.The median cumulative survival time of patients with HCC was 48 months,which was significantly longer than 19 months of patients with CC and 14 months of patients with cHCC-CC (Log-rank value =4.354,8.847,P ＜ 0.05).The median tumor-free survival time of patients with HCC was 26 months,which was significantly longer than 9 months of patients with CC and 9 months of patients with cHCC-CC (Log-rank value =6.479,7.708,P ＜ 0.05).The tumor recurrence rate within 1 year of patients with HCC was 28.8％ (152/527),which was significantly lower than 57.7％ (15/26) of patients with CC or 9/14 of patients with cHCC-CC (F =17.046,P ＜ 0.05).No vascular thrombosis was detected in patients with CC,but the regional lymph node metastasis rate was 19.2％ (5/26),which was significantly higher than 2.8％ (15/527) of patients with HCC (x2 =19.082,P ＜ 0.05).Level of alpha-fetoprotein,TNM staging,tumor diameter,multiple foci,liver capsule invasion,satellite foci and lymph node metastasis were risk factors for the survivals of patients with primary liver cancer after hepateetomy (x2 =8.648,118.786,59.548,7.639,13.200,43.842,15.540,P ＜ 0.05).Vascular tumor thrombosis and Child-Pugh classification were the risk factors for the survivals of patients with HCC or cHCC-CC (x2 =70.446,6.230,P ＜ 0.05).TNM staging,tumor diameter,satellite foci and vascular tumor thrombusis were the independent risk factors for the survivals of patients with primary liver cancer (RR =1.420,1.050,1.513,1.899,P ＜ 0.05) ; TNM staging,tumor diameter and vascular tumor thrombosis were the independent risk factors for the survivals of patients with HCC (RR =1.432,1.888,1.052,P ＜ 0.05).TNM staging and tumor diameter were the independent risk factors for the survivals patients with CC (RR =1.473,1.503,P ＜ 0.05).Conclusion Although CC and cHCC-CC take small proportion in the primary liver cancer,the tumor recurrence rate is higher and the survival rate is lower when compared with patients with HCC.

The zinc-regulated transporters (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) plays an important role in the growth and development of plant. In this study, a full length cDNA of ZIP encoding gene, designed as DoZIP1 (GenBank accession KJ946203), was identified from Dendrobium officinale using RT-PCR and RACE. Bioinformatics analysis showed that DoZIP1 consisted of a 1,056 bp open reading frame (ORF) encoded a 351-aa protein with a molecular weight of 37.57 kDa and an isoelectric point (pI) of 6.09. The deduced DoZIP1 protein contained the conserved ZIP domain, and its secondary structure was composed of 50.71% alpha helix, 11.11% extended strand, 36.18% random coil, and beta turn 1.99%. DoZIP1 protein exhibited a signal peptide and eight transmembrane domains, presumably locating in cell membrane. The amino acid sequence had high homology with ZIP proteins from Arabidopsis, alfalfa and rice. A phylogenetic tree analysis demonstrated that DoZIP1 was closely related to AtZIP10 and OsZIP3, and they were clustered into one clade. Real time quantitative PCR analysis demonstrated that the transcription level of DoZIP1 in D. officinale roots was the highest (4.19 fold higher than that of stems), followed by that of leaves (1.12 fold). Molecular characters of DoZIP1 will be useful for further functional determination of the gene involving in the growth and development of D. officinale.
Amino Acid Sequence ; Cloning, Molecular ; Dendrobium ; chemistry ; classification ; genetics ; metabolism ; Gene Expression Regulation, Plant ; Iron ; metabolism ; Membrane Transport Proteins ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Plants ; chemistry ; classification ; genetics ; Sequence Alignment ; Zinc ; metabolism