Objective To evaluate the accuracy of performance of Engstrom Eas 9010 vaporizerMethods Using Datex Capnomac AS/3 monitored the output of Gambro-Engstrom Eas 9010 anesthetic agent vaporizer under various situations l Determing the output of vaporizer at different gas flow rates of 0 5,1 0,2 0,4 0,6 0 and 8 0 L?min -1 2 The effects resulting from up stream oxygen flushing 3 The effects due to additional nitrous oxide as carrier gas 4 Effects of different airway pressure at the same flow rate Results 1 The output of the vaporizer was highly correlated with the dial setting at various gas flow rate except dial setting was more than 8% and gas flow rate less than 0 5L?min -1 2 Upstream oxygen flush could not decrease the concentration remarkably 3 The output of the vaporizer was not affected by additional carrier gas 4 The effect on vaporizer output was not remarkable under different airway pressure at the same flow rate Conclusions The anesthetic vaporizer principle used in the Gambro-Engstrom Eas 9010 apparatus is innovative Electronically controlled vapor quanta injection is used with safety feed back Its accuracy can't be influenced by temperature,pressure and carrier gas indicating to be safely applied to the low or even minimal flow anesthesia

Objective To summarize the anatomical variations of the Calot's triangle and explore the best method to manage the variations during laparoscopic cholecystectomy(LC).Methods From December 2006 to December 2008,158 patients with anatomical variation of the Calot's triangle received LC,the clinical data of the cases were reviewed retrospectively.Results Among the cases,15 patients were converted to open surgery because of Ⅰ type Mirizzi syndrome(3 cases),Ⅱ type Mirizzi syndrome(4 cases),low location of the convergence of the cystic duct and the common bile duct(2 cases),cystic duct opening into the posterior wall of the common bile duct(2 cases),the cystic duct and common bile duct sharing 2-cm lateral wall(1 case),severe adhesion of the Calot's triangle(2 cases),and hemorrhage of the posterior cystic artery(1 case).The LC were completed in 143 patients,among which 5 cases had postoperative complications,including biliary leakage in 1 case(cured by a second operation),bleeding at the puncture sites in 2 patients,infection of the puncture site in 1 case,and residual cystic stones in 1 case(cured by ERCP in 2 weeks).Conclusions Knowledge of the anatomical variations of the Calot's triangle is the key to LC.Different surgical strategies should be carried out according to the dissection of the Calot's Triangle area,and the location of the common hepatic duct and common bile duct.

The interaction of drug and target protein is a critical part of new drug discovery. It is the premise for drugs to exert therapeutic effects by targeting specific binding sites of target proteins and thereby affecting its pharmacological activity. Currently, a variety of techniques are exploited to detect the interaction between drug ligands and target proteins. For example, cellular thermal shift assay (CETSA) and differential scanning fluorimetry (DSF) based on thermodynamics, mass spectrometry and nuclear magnetic resonance technology, etc. In addition, high-throughput ligand screening technology provides technical convenience for the search of specific ligand, and is a powerful tool to efficiently identify the interaction between drug ligand and target protein. Here, we summarize the detection techniques of interaction between small molecules and target proteins, and discuss the application of high-throughput ligand screening technology in drug research.

Target identification and verification of natural products is an important and challenging work in the field of chemical biology. It is also an important job for researchers to apply chemical proteomics technology to biomedicine in order to identify target proteins of natural products. Target identification is critical to understanding its mechanisms and developing natural products as molecular probes and potential therapeutic drugs. Traditional approaches of small molecule target identification based on affinity have been shown to be successful, such as click-chemical probes, radioisotope labeling or photosensitized small-molecule probes. Nevertheless, these technologies require purified candidate target proteins, and modified small molecules with probes or linkers, such as adding agarose beads, biotin labels, fluorescent labeling or photo-affinity labeling. Many structure-activity relationship studies should be performed to ensure that the addition of small molecule labels undisturbed the original biological activity of the small molecules. Unfortunately, all these modifications are likely to alter their biological activity or binding specificity. To overcome the bottleneck of "target recognition", researchers have developed a series of new techniques for unmodified drug target identification. In this article, we reviewed the target identification techniques of natural product without structural modification in order to provide reference for the development of natural products.

Objective:To screen for the pathogenesis-related genes of osteosarcoma and to assess their roles for the de- velopment of osteosareoma.Methods:Total RNA was extracted from 3 ATCC osteosarcoma cell lines and an osteoblastic cell line and was used to synthesize biotinylated cRNAs;the latter were hybridized to Affymetrix~(?)GeneChip~(?)U133A ar- rays and a gene with more than 2 folds of change was selected.Ten of the differentially expressed genes were chosen and the primers were designed and the synthesized.Then SYBR~(?)Green real-time PCR(RT-PCR)method was used to detect the expression of the 10 genes in 9 fresh osteosarcoma specimens.ABI Prism 7 000 system was used to analyze the differ- ent expression between osteosarcoma cell line and osteoblastic cell line.Results:We identified 58 up-regulated and 142 down-regulated genes in the 3 osteosareoma cell lines.Many of the genes were firstly reported to be related to the patho- genesis of osteosarcoma.These differentially expressed genes were mainly involved in energy and material metabolism,on- cogene,signal transduction gene,transcription- related genes,cell cycle-related genes,cell apoptosis-related gene,im- mune response gene,tumor suppressor genes,etc.The array results of 10 randomly selected genes were further verified by the RT-PCR in 9 fresh osteosarcoma specimens.Conclusion:Many genes are involved in the pathogenesis of osteosarcoma. Gene microarray can help to discover the genes related to the pathogenesis of osteosarcoma,which may lay a foundation for studying the molecular mechanism of osteosarcom.

OBJECTIVETo observe the expression of PECAM-1 and E-selectin in the vulnerable plagues and their relationships to myocardial Leu125Val polymorphism of PECAM-1 and Ser128Arg polymorphism of E-selectin in autopsied samples of patients with acute coronary syndrome (ACS).

METHODSWe detected the expressions of PECAM-1 and E-selectin in the vulnerable plaques by immunohistochemistry on 50 autopsy samples of patients with ACS, 30 autopsy samples from non-cardiac disease patients served as control. Genetic Leu125Val polymorphism of PECAM-1 was detected by PCR-SSCP in myocardial paraffin blocks of 37 ACS cases and 43 control cases, and Ser128Arg polymorphism of E-selectin was detected by PCR-RFLP in myocardial paraffin blocks of 39 ACS cases and 43 control cases, respectively.

RESULTSImmunohistochemical features: (1) the incidence of positive expression in the intima of coronary artery of PECAM-1 [76.0% (38/50) vs. 26.7% (8/30)] and E-selectin [26.0% (13/50) vs. 0] was significantly higher in ACS group than in control group (all P < 0.01). (2) Expressions of PECAM-1 [58.0% (29/50) vs. 28.0% (14/50)] and E-selectin [22.0% (11/50) vs.12.0% (6/50)] were significantly higher at neovascular endothelial cells in plaques than expressions at coronary arterial endothelial cells in ACS group (all P < 0.01). (3) In 41 plaques with inflammatory infiltration, the expression rates of PECAM-1 and E-selectin in inflammatory cell density of < 10, 10 - 30 and > 30/HPF were 33.3%, 68.2%, 92.3% and 16.7%, 31.8% and 23.1%, respectively. Genotype detection results: There is significant difference in frequencies of allele in Leu125Val polymorphism (P < 0.05), but the genotype distributional frequencies were similar (P > 0.05) between ACS group and control group. There are significant differences in frequencies of allele and genotype in Ser128Arg of E-selectin polymorphism between ACS group and control group (all P < 0.05).

CONCLUSIONSThe immunohistochemical expressions of PECAM-1 and E-selectin were significantly increased at intima in vulnerable plaques of ACS group, especially in neovascular endothelial cells, and positively correlated with inflammatory cell density, suggesting that PECAM-1 and E-selectin might play an important role in inflammatory reaction and development of vulnerable plaque. E-selectin Ser128Arg polymorphism is associated with ACS, and it might be a risk factor for ACS.

Acute Coronary Syndrome ; genetics ; metabolism ; Aged ; Aged, 80 and over ; E-Selectin ; genetics ; metabolism ; Female ; Gene Frequency ; Genotype ; Humans ; Inflammation ; Male ; Middle Aged ; Myocardium ; metabolism ; Platelet Endothelial Cell Adhesion Molecule-1 ; genetics ; metabolism ; Polymorphism, Single Nucleotide ; Risk Factors

Objective To observe the expression level of inhibitor of differentiation 2 (Id‐2) and matrix metalloproteinases‐9 (MMP‐9) in rectal cancer ,analysis the correlation of the expression level of them ,to study the relationship between the expression level of them and the clinical pathology indicators of rectal cancer .Methods Rectal cancer tissues and normal tissue adjacent to rec‐tal cancer were obtained from the rectal cancer resection of 56 patients with rectal cancer ,using immunohistochemical method to ob‐serve the expression level of Id‐2 and MMP‐9 in normal tissue adjacent to rectal cancer and rectal cancer and Spearman correlation test to detect the correlation between the expression level of Id‐2 and MMP‐9 ;then we analyzed the relationships between the ex‐pression level of Id‐2 and MMP‐9 and the index of rectal cancer clinical pathology .Results The positive expression rate of Id‐2 in the in rectal cancer tissues is more higher than that of normal tissue of adjacent to rectal cancer (73 .21% vs .48 .21% ,P<0 .05) . The positive expression rate of MMP‐9 in the in rectal cancer tissues is higher than that of normal tissue of adjacent to rectal cancer (71 .43% vs .44 .64% ,P<0 .05) .Spearman correlation test showed that there is the positive correlation between the expression level of Id‐2 and MMP‐9 (r=0 .393 ,P=0 .003) .The expression levels of Id‐2 and MMP‐9 in rectal cancer were correlated with the degree of tumor differentiation ,TNM stage and lymph node metastasis (P<0 .05) ,but had no differences between the elements of age and sex (P>0 .05) .Conclusion There is a close relationship between the expression levels of Id‐2 and MMP‐9 in rectal cancer and the occurrence and development of rectal cancer .Rectal cancer with the higher Id‐2 expression level may be the ways to achieve tumor invasion and metastasis through MMP‐9 as a facilitator .

Objective To explore the surgical modality and safety of hepatectomy for hepatic hemangiomas close adjoining the hepatic portal and vital blood vessels. Methods From June 2005 to June 2010 17patients of hepatic hemangiomas underwent hepatectomy. Data were retrospectively analyzed.Results All the 17 cases were operated on successfully. Six cases were treated with anatomic liver lobectomy including right hemibepatectomy through liver hanging maneuver by anterior approach in 2 cases,under right liver blood vessel blochade and anatomic right posterior hepatectomy in 2 cases, left hemihepatectomy in 2 cases. Eight cases were treated by hemangiomas enucleation, in 3 cases hemangioma was enuleated through liver parenchyma splitting under intermittent hepatic blood inflow exclusion. There was no postoperative mortality, postoperatively pleural effussion occured in 5 cases,wound infection occured in 1 case, and pulmonary infection occured in 1 case, all the cases were cured. Conclusions Different operation styles should be applied according to the position, size of hepatic hemangiomas close adjoining the hepatic portal and the important blood vessels.

Objective To clone and express the encoding sequence of glyceraldehydes-3-phosphate dehydrogenase(GAPDH)from periodic Brugia molayi(Bm).Methods Total RNA was extraeted from periodic Brugic malayi.The BmGAPDH gene was amplified by RT-PCR.The PCR product was cloned and then subeloned into pcDNA3.1(+)vector.The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification,and were transformed into COS-7 cell subsequently.The expressed protein was identified by SDS-PAGE.Results BmGAPDH mRNA was highiy expressed in transfected COS-7 cell.The deduced amino acid sequence was identical with that of BmGAPDH.The recombinant pnotein wag about Nr 43 000.Conclusion The recombinant plasmid peDNA3.1(+)-BmGAPDH has been constructed and the protein has been expressed correctly.

OBJECTIVETo economically obtain the Abeta peptide for Alzheimer's disease (AD) research by expressing the Abeta peptide fused with the maltose binding protein (MBP) possessing high solubility in E.coli.

METHODSThe cDNA-coding sequence of Abeta peptide was modified by the addition of a BamH I site at the 5' end and a Hind III site at the 3' end using PCR. The modified sequence was ligated into the maltose-binding protein (MBP) fusion expression vector pMAL-c2 containing an thrombin cleavage site, which was transformed into competent E.coli DH5alpha cells. After identification of the single clones by PCR and DNA sequencing, the recombinant plasmid was transformed into E.coli TB1 and induced to express MBP-Abeta fusion protein. The expressed fusion protein was purified using amylose resin column and identified by SDS-PAGE and Western blotting.

RESULTSThe result of DNA sequencing verified the consistency between the inserted sequence and Abeta (1-42) sequence. SDS-PAGE electrophoresis showed that MBP-Abeta fusion protein was highly expressed in E.coli TB1, and Western blotting demonstrated that the purified fusion protein and the separated Abeta peptide could be recognized by specific anti-Abeta (22-35) antibody.

CONCLUSIONMBP-Abeta fusion protein highly expressed in E. coli TB1 cells with enhanced solubility and the separated Abeta peptide with good immunogenicity obtained may lend support to AD research.

Alzheimer Disease ; genetics ; Amyloid beta-Peptides ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Maltose-Binding Proteins ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification