Novozym 435 was selected from four lipases and two proteinase because of its high catalytic activity and enantiosectivity.For the ammonolysis of (?)-?-methylbenzyl acetate,The effect of ammonia sources,the concentration of enzyme and substrates on the reaction were further explored .under the optimum conditions of this study,after 6h reaction,with the enantiomeric excess of the remaining (-)-?-methylbenzyl acetate was found to be higher than 99%.

The effects of reaction media, water activity, temperature and pH on Novozym 435-catalyzed enantiose-lective ammonolysis of (?) -?-methylbenzyl acetate have been systematically explored. Novozym 435 showed high catalytic activity and enantioselectivity in hexane; the optimum temperature and the initial water activity were 25℃ and 0.33 respectively; The suitable reaction pH was in the range of 6.0 - 7.0.

Objective To evaluate the clinical application of CT-guided localization with combination of methylene blue and a Hookwire system for small pulmonary nodules (SPNs) before video-assisted thoracoscopic resection.Methods CTguided localization the SPNs before resection in 56 patients and 60 nodules,then underwent video-assisted thoracic surgery (VATS) resection.Among 56 patients,19 males and 37 females,aged from 35 to 81 years,mean age was (61.1 ±8.9)years.Results SPNs diameter (6.80 ±4.12) mm,distance from the parietal pleura (15.38 ±4.63) mm.CT-guided localization success rate was 100％,positioning time (10.76 ± 8.17) min,8.9％ (5/56) had micro pneumothorax aftet positioning,7.1％ (4/56) occurrence of needle tract bleeding,no conservative treatment.VATS resection rate was 100％.The pathology of 60 lesions were shown:Bronchiolo-alveolar carcinoma(BAC) were 33 lesions(55.0％),BAC and adenocarcinoma were 11 lesions(18.3％),Atypicaladenomatous hyperplasia (AAH) were 7 lesions (11.8％),Inflammation were 4 lesions (6.7％),Harmatoma were 3 lesions(5.0％),Tuberculoses were 2 lesions(3.3％).Conclusion CT-guided localization with combination of methylene blue and a Hookwire system before video-assisted thoracoscopic resection is a promising technique for small solitary pulmonary nodules.It could play an important role in accurate localization of small pulmonary nodules,and it is a safe technique with clinical application.

A complete aniline dioxygenase gene cluster cloned from an Acinetobacter sp. strain, which could utilize aniline as the sole carbon, nitrogen and energy, was sequenced. Sequence analysis showed that the gene cluster had six intact ORFs, and the whole sequence had high similarity with that of Acinetobacter sp. YAA at amino acid level. A recombinant strain was formed with the gene cluster ligated to vector pLAFR6 and transferred to E.coli. After optimizing the fermentation conditions of this strain for producing catechol, LB was confirmed as the final medium, pH7.0, aniline concentration 0.5mg/ml, E.coli DH5?as the host, incubation temperature 37℃, amount of inoculum 3%. Under above conditions, the yield of catechol could get to 0.546mg/ml, and the converting rate of substrate at molecule level could get to 92.4%.

OBJECTIVETo evaluate whether lipid-lowering therapy with Xuezhikang can reduce the risk of cardiac events and total mortality in coronary heart disease (CHD) patients with or without hypertension.

METHODSIn this random, double-blinded, placebo controlled clinical trial, 2704 patients with hypertension and 2166 patients without hypertension were enrolled and capsule Xuezhikang 0.6 g Bid or placebo on the top of conventional therapy without other lipid-lowering drugs. The mean follow-up period was four years. The primary end-points were nonfatal myocardial infarction and total mortality.

RESULTSCompared to placebo group, the incidence of cardiac events was reduced by 44.0% (P<0.0001) and 47.4% (P<0.0001) respectively in CHD patients with or without hypertension, and the total mortality was lowered by 35.8% (P=0.0012) and 28.6% (P=0.0737) respectively in CHD patients with or without hypertension. There was no significant difference in side effects between study groups.

CONCLUSIONXuezhikang can reduce the cardiac events and mortality in CHD patients with or without hypertension.

Adolescent ; Adult ; Aged ; Coronary Disease ; drug therapy ; mortality ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Hypertension ; complications ; drug therapy ; mortality ; Hypolipidemic Agents ; therapeutic use ; Lipids ; blood ; Middle Aged ; Phytotherapy ; Survival Rate

OBJECTIVETo detect the sequence variations frequently found within the N- and C-terminal regions of Epstein-Barr virus (EBV) LMP1 gene in nasopharyngeal carcinoma (NPC) and to study the underlying mechanisms.

METHODSFresh tumor tissues were sampled from 63 patients with untreated NPC encountered in Affiliated Tumor Hospital of Sun Yat-sen University, Guangzhou. The N-terminal region of EBV LMP1 gene was amplified with nested polymerase chain reaction (PCR), followed by XhoI enzyme digestion. Nested PCR was also employed to detect the 30 base pairs deletion within the C-terminal region. Four-colored fluorescence terminator sequencing method was applied for bi-directional solid-phase sequencing of the 8 representative PCR products in 4 cases of NPC. The DNA sequence within the N- and C-terminal regions of LMP1 gene was then analyzed.

RESULTSThere were 4 patterns of sequence variations, namely, wt-XhoI/wt-LMP1 (4 cases, 6.3%), wt-XhoI and XhoI-loss/del-LMP1 (4 cases, 6.3%), wt-XhoI/del-LMP1 (5 cases, 7.9%) and XhoI-loss/del-LMP1 (50 cases, 79.5%), detected in the 63 studied cases. Sequence analysis showed that the EBV LMP1 gene had underwent non-synonymous and synonymous substitutions, as compared with the prototype of B95-8 cells. The ratio of non-synonymous to synonymous substitutions was 2.25.

CONCLUSIONSXhoI-loss/del-LMP1 is the predominant sequence variation pattern of EBV LMP1 gene in NPC from Guangzhou. The XhoI-loss variation seems to develop on top of del-LMP1. When compared with the EBV LMP1 gene in peripheral blood B-lymphocytes of virus carriers and in preinvasive epithelial lesions (reported previously), it is likely that the sequence variation patterns of LMP1 gene may represent 4 different phases of intrahost evolution of EBV during nasopharyngeal carcinogenesis.

Adult ; Aged ; Base Sequence ; DNA, Viral ; genetics ; Deoxyribonucleases, Type II Site-Specific ; genetics ; Female ; Gene Deletion ; Genetic Variation ; Herpesvirus 4, Human ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Nasopharyngeal Neoplasms ; virology ; Point Mutation ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

OBJECTIVETo study the subcellular localization of human endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) protein in endothelial cells.

METHODSHuman umbilical vein endothelial cell strain ECV304 were cultured in vitro. The fusion protein of enhanced green fluorescent protein (EGFP)-EOLA1 expressing plasmid was constructed. Empty plasmid with EGFP at N side (pEGFP-N2) and fusion protein expressing plasmid EGFP-EOLA1 was respectively transfected into ECV304 cells with liposome. After being cultured for 48 hours, the expression levels of EGFP and fusion protein EGFP-EOLA1 in cells were detected with Western blot. The subcellular localization of EOLA1 protein was detected by laser scanning confocal microscope and immunoelectron microscopy.

RESULTSThe EGFP-EOLA1 coexpression plasmid was verified to be successfully constructed by enzyme cutting and gene sequencing. The fusion protein of EGFP-EOLA1 was observed to express in transfected cells through Western blot. Green fluorescence scattered all over the ECV304 cells transfected with empty plasmid and cells transfected with fusion protein expressing plasmid, and it gathered obviously in the nuclei in the latter cells. Immune deposits were observed in the matrix of cells transfected with fusion protein expressing plasmid but not in the cells transfected with empty plasmid.

CONCLUSIONSEOLA1 protein is localized in the nucleus and the matrix of ECV304 cell, and it plays its role as a signal transduction factor.

Cell Line ; Cell Nucleus ; metabolism ; Human Umbilical Vein Endothelial Cells ; metabolism ; Humans ; Lipopolysaccharides ; metabolism ; Membrane Proteins ; metabolism ; Signal Transduction

Anti-Inflammatory Agents ; pharmacology ; Capsules ; China ; Coronary Disease ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Humans

OBJECTIVETo investigate the clinicopathologic characteristics of primary nasopharyngeal adenoid cystic carcinoma (NPACC) and its relation to Epstein-Barr virus (EBV) infection in Guangzhou where is a high-incidence area of EBV-associated nasopharyngeal carcinoma (NPC).

METHODS17 cases of NPACC with clinical record and biopsy samples were collected in Guangzhou and their clinical manifestations were reviewed. Besides HE, Alcian blue and PAS, LSAB immunohistochemistry was performed for detecting the expression of a variety of epithelial markers, CD21 and EBV encoded LMP1. EBV encoded early RNAs (EBER) was detected by using in-situ hybridization. Nested PCR was applied for studying the presence of EBV W-fragment in tissues.

RESULTSThe ratio of male to female was 7:10. The patients' age ranged from 30 to 63 years, and the median age was 46 years. 14 out of 17 tumors showed markedly local aggressive growth, presenting as T3 or T4. However, only 1 patient had metastasis of an ipsilateral cervical lymph node. The majority of neoplastic cells were basal-cell like in shape and with scanty cytoplasm and a deeply stained nucleus. Intercellular hyaline or mucinous substance was always present in between the carcinoma cells. Cribriform structure formed by the neoplastic cells could be found in 16 out of these 17 biopsies. The NPACC always express the wide-spectrum cytokeratin and the epithelium membrane antigen. Only a few or a small number of carcinoma cells showed nuclear EBER-signals in 9 cases (9/17). Concurrently, these 9 NPACCs showed a 192 bp W-fragment positive band on electrophoresis gel by nested PCR. LMP1 expression had been found in 5 out of the 9 NPACCs (55.6%) accompanying with EBER-positive carcinoma cells. The EBER-positive infiltrating lymphocytes could also be found in the stroma of 3 out of the 9 EBER-stained NPACC slides. All the tumor cells, including the EBER-positive cell of the 17 NPACCs showed no CD21 expression.

CONCLUSIONSThe female is predominant over the male in development of NPACC, which often accompanied with a markedly invasive capacity at the nasopharynx and its neighboring sites. Only a small number of tumor cells, nearly a half of the studied cases were infected with EBV. Therefore, it's postulated that there seems no close relation present between NPACC and EBV infection.

Adult ; Carcinoma, Adenoid Cystic ; etiology ; pathology ; Epstein-Barr Virus Infections ; complications ; Female ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; etiology ; pathology ; RNA, Viral ; analysis ; Viral Matrix Proteins ; analysis

Blotting, Western ; DNA, Ribosomal ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; biosynthesis ; Transfection ; Vascular Endothelial Growth Factor A ; genetics