The gene coding for beta-glycosidase (EC3.2.1.21) from Thermus nonproteolyticus HG102 has been cloned and expressed in E. coli. The gene open reading frame was 1311 bp and it codes for 436 amino acids. The deduced amino acid sequence of the enzyme showed identity with members of glycosyl hydrolase family I. The enzyme had high content of hydrophobic amino acid (Ala 12.8%, Leu 10.9%), Arg(9.6%), Glu(9.4%) and Pro(8.0%), but low content Cys(0.45%) and Met (0.9%). From the alignment of enzyme amino acid sequence with other glycosyl hydrolase family I members, Glu164 and Glu338 were predicated as the proton donor and nucleophile group. The DNASTAR program was used to predict the secondary structure. According to the Chou-Fasman model, the enzyme has 41.4% of alpha-helics, 16.2%, beta-strands, 14.4%, beta-turns. 14 of the 35 Pro were located at the second sites of beta-turns. Hydrophobic interaction, ion bond, alpha-helics and Pro had important contribution to Tn-gly thermostability.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; Glycoside Hydrolases ; biosynthesis ; classification ; genetics ; Hot Temperature ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Phylogeny ; Protein Structure, Secondary ; physiology ; Recombinant Proteins ; biosynthesis ; genetics ; Sequence Analysis, DNA ; methods ; Sequence Homology ; Thermus ; enzymology ; beta-Glucosidase

To automatically infer the patterns of vessel structure such as the distal ends, segments, bifurvessel structures, and crossing of two vessels in X-ray angiographic images, a novel method is presented based on Gabor filter and circle detector. The method can cope with varying vessel curvature and intensity feature occur along the longitudinal vessel direction. The present study can facilitate 2-D quantitative description of vessel tree and 3-D vessel reconstruction, and provide an elementary clue for the diagnostics. The proposed method has been successively applied to both synthetic images for validation purposes and the actual angiographic images, which yielded encouraging results.
Algorithms ; Angiography ; methods ; Artifacts ; Blood Vessels ; anatomy & histology ; pathology ; Humans ; Image Enhancement ; methods ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Pattern Recognition, Automated ; methods

This study aimed to evaluate the vasorelaxant effect and mechanisms of compound reserpine and triamterene tablets (CRTTs) and its component triamterene on isolated rat thoracic aorta rings. Isolated rat thoracic aorta rings pre-contracted by high potassium or norepinephrine (NE) were used to evaluate the vasodilatory effect of CRTTs and its component triamterene. The mechanisms concerning endothelium, potassium channels and calcium channels were studied through the interventions of several tool drugs. Animal welfare and experimental procedures followed the requirements of the Laboratory Animal Management and Animal Welfare Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College. The results showed that both CRTTs and triamterene had potent relaxant effect on KCl and NE pre-contracted vessels. Triamterene showed partial endothelium dependency, and N-nitro-L-argininemethyl ester hydrochloride (L-NAME) could influence the relaxant effect, which may be related to the opening of calcium-activated potassium channels. Meanwhile, CRTTs could inhibit intracellular Ca²⁺ release and extracellular Ca²⁺ influx. Overall, CRTTs and its component triamterene have vasorelaxant effects on vessels in vitro, and the vasorelaxant effect of CRTTs may be related to intracellular Ca²⁺ release and extracellular Ca²⁺ influx, while this effect of triamterene may depend on vascular endothelial function and may also be related to the opening of calcium-activated potassium channels.

Objective To study the expressions of nucleostemin gene mRNA and Ki-67antigen in pituitary adenomas and investigate the role of nucleostemin gene in the tumorigenesis and development of pituitary adenomas. Methods Seventy-one samples of pituitary adenomas were collected. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression of nucleostemin gene mRNA in samples. Immunohistochemistry technique was applied to examine Ki-67 antigen expression in the paraffin sections of samples. At the same time,coherent clinical data were collected. Results Nucleostemin gene mRNA was detectable in all samples of pituitary adenomas. Between invasive and noninvasive pituitary adenomas, the difference of nucleostemin expression was extremely significant (P＜0.01), and Ki-67 labeling index (LI) was also significantly different (P＜0.05). The difference of Ki-67LI between recurrent and non-recurrent groups was significant (P＜0.05). There was a positive correlation between nucleostemin gene and Ki-67LIlevels (r=0.237, P＜0.05). Conclusions Nucleostemin gene plays an important roles in the invasion of human pituitary adenoma. Expression of nucleostemin gene is positively related to Ki-67 antigen expression, and both may be the valid clinical detection markers for assessing proliferation, invasion and recurrence of pituitary adenomas.

By analyzing the design principles and treatment modes of Gambro Prismaflex CRRT device,based on the basic structure and treatment process,and the typical failure cases of pressure joints and scale zeroing test failures in continuous renal replacement therapy(CRRT)equipment were analyzed,the targeted solution and maintenance strategies were proposed to ensure the stable and efficient operation of CRRT equipment.

A portable ECG monitor is introduced in the paper, which has a temporary intravenous and transesophageal fixable rate pacing function. During the PSVT attack, the tachyarrythmia can be stopped by the transesophageal cardiac pacing while the ECG signals are monitored. The instrument has some advantages such as small size, low price and good practicality. It is worth while introducing and popularizing it to all hospitals.
Electrocardiography, Ambulatory ; instrumentation ; Equipment Design ; Heart Block ; diagnosis ; therapy ; Heart Rate ; Humans ; Monitoring, Ambulatory ; instrumentation ; Pacemaker, Artificial ; classification ; Signal Processing, Computer-Assisted ; Software ; Tachycardia ; diagnosis ; therapy

OBJECTIVETo investigate the inhibitory effect of Oridonin injection on heterotransplanted tumors of human gastric adenocarcinoma cell line BGC823 cells in nude mice and explore its mechanism.

METHODSHeterotransplanted models of human gastric adenocarcinoma cell line BGC823 cells in nude mice were established. They were divided at random into three groups as control group, low-dose group and high-dose group. The Oridonin solution at concentration of 37.5 mg x kg(-1 x d(-1) and 75 mg x kg(-1) x d(-1) were injected to the mice in low-dose group and high-dose group, respectively, and 0.9% sodium chloride was injected to the mice of control group per day for 10 days sequentially. The mice of the three groups were sacrificed at 11th day after the first injection of Oridonin. The tumor weight of the sacrificed mice was measured. Morphological and ultrastructural examinations of the tumors were carried out by light and electron microscopy. The expression of bcl-2, Bax, Fas and FasL was detected by immunohistochemistry.

RESULTSOridonin injection showed a suppressive effect on the growth of heterotransplanted tumors in the nude mice. The tumor growth inhibition rates were 48.5% and 70.7% in the low-dose and high-dose groups, respectively. The morphological study demonstrated that tumor cells displayed a typical appearance of apoptosis. The expression of bcl-2 was down-regulated, while Bax, Fas and FasL were up-regulated.

CONCLUSIONOridonin can markedly inhibit the growth of heterotransplanted human gastric adenocarcinoma in nude mice. It was due, at least in part, to the induction of apoptosis in cancer cells.

Adenocarcinoma ; metabolism ; pathology ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Diterpenes, Kaurane ; administration & dosage ; pharmacology ; Dose-Response Relationship, Drug ; Fas Ligand Protein ; metabolism ; Humans ; Injections ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Stomach Neoplasms ; metabolism ; pathology ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism

OBJECTIVETo assess the optimal regimen and its mechanism of ZD1839 in combination with SN38, the active metabolite of irinotecan (CPT-11), in the colon cancer cell lines HT-29 and LoVo.

METHODSChou and Talalay method was used to analyze the combination effects of sequencing of ZD1839 and SN38. Western blotting and immunoprecipitation were used to determine the effects of ZD1839 and/or SN38 on their targeted enzymes and downstream markers. Apoptosis was assayed by analyzing histone-associated DNA fragment.

RESULTSSequential SN38 followed by ZD1839 produced a synergistic effect. In contrast, SN38 following ZD1839 exhibited an antagonist effect. SN38 markedly inhibited topoisomerase I (Topo-I) activity. ZD1839 did not alter epidermal growth factor receptor (EGFR) expression, but resulted in a complete inhibition of EGFR phosphorylation. Sequential ZD1839 followed by SN38 did not show any enhanced inhibition effect on Topo-I activity, phosphorylation of EGFR and one of its downstream markers MAPK. However, simultaneous SN38 plus ZD1839, and sequential SN38 followed by ZD1839 administrations showed modest inhibition effect on EGFR's another downstream marker AKT. The combination schedules also showed prominent influence on cell cycle distribution. ZD1839 maintained SN38-induced DNA damage and apoptosis.

CONCLUSIONSequential SN38 followed by ZD1839 may be a favorable combination schedule.

Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Camptothecin ; analogs & derivatives ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Colonic Neoplasms ; enzymology ; metabolism ; pathology ; DNA Topoisomerases, Type I ; metabolism ; Drug Synergism ; HT29 Cells ; Humans ; Inhibitory Concentration 50 ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Phosphorylation ; drug effects ; Proto-Oncogene Proteins c-akt ; metabolism ; Quinazolines ; pharmacology ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; metabolism ; Signal Transduction ; drug effects

OBJECTIVETo identify the mutation of a Chinese family with inherited long QT syndrome(LQTS).

METHODSThe disease-causing gene was tentatively determined in light of the clinical manifestations and electrophysiological properties, and then polymerase chain reaction and DNA sequencing were used for screening and identifying mutation.

RESULTSA missense mutation G940A(G314S) in the KCNQ1 gene was identified, which was the 'hot spot' of long QT syndrome mutation.

CONCLUSIONThe mutation that is involved with long QT syndrome in Chinese patients is the same as that in the European, American and Japanese patients.

China ; DNA Mutational Analysis ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; KCNQ1 Potassium Channel ; genetics ; Long QT Syndrome ; diagnosis ; genetics ; Male ; Mutation, Missense ; Pedigree ; Polymerase Chain Reaction

OBJECTIVETo investigate the molecular pathology in families with long QT syndrome (LQTS) including Jervell-Longe-Nielsen syndrome (JLNS) and Romano-ward syndrome (RWS) and Brugada syndrome (BS) in Chinese population.

METHODSPolymerase chain reaction and DNA sequencing were used to screen for KCNQ1, KCNH2, KCNE1, and SCN5A mutation.

RESULTSWe identified a novel mutation N1774S in the SCN5A gene of the BS family, a novel mutation G314S in a RWS family which had also been found in Europe, North America, and Japan, and a single nucleotide polymorphisms (SNPs) G643S in the KCNQ1 of the JLNS family. In this JLNS family, another heterozygous novel mutation in exon 2a was found in KCNQ1 of the patients.

CONCLUSIONNew mutations were found in our experiment, which expand the spectrum of KCNQ1 and SCN5A mutations that cause LQTS and BS.

Adolescent ; Adult ; Base Sequence ; ERG1 Potassium Channel ; Ether-A-Go-Go Potassium Channels ; genetics ; Female ; Humans ; Jervell-Lange Nielsen Syndrome ; genetics ; KCNQ1 Potassium Channel ; genetics ; Long QT Syndrome ; congenital ; genetics ; Male ; Middle Aged ; Molecular Sequence Data ; Muscle Proteins ; genetics ; Mutation ; NAV1.5 Voltage-Gated Sodium Channel ; Pedigree ; Potassium Channels, Voltage-Gated ; genetics ; Romano-Ward Syndrome ; genetics ; Sodium Channels ; genetics