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Biomedical subjects

Yang Yang

Publications and source records attributed to Yang Yang.

At least 163 records · Page 9Linked to original sources
PubMed2004

[Study of zinc in protecting sperm from sodium nitroprusside damage].

OBJECTIVE: To study the function of zinc in preventing human sperm from being damaged by sodium nitroprusside (SNP), an external NO donor. METHODS: Analyses were made of the function of zinc in protecting sperm from being influenced by SNP in such aspects as sperm motility, head-tail connection and the breakage of sperm DNA chain by using phase-contrast microscope and single cell gel electrophoresis (SCGE). RESULTS: Sperm motility was obviously inhibited by SNP. The percentage of comet cells increased significantly but the stability of sperm head-tail connection decreased. Zinc could promote sperm motility, protect the DNA chain and prevent the sperm head-tail connection from breaking. CONCLUSION: Zinc can protect sperm from being damaged by NO. Its mechanism may be related to the mercaptol group of sperm chromatin.

Adult↗
PubMed2004

[Site-directed mutagenesis and promoter functional analysis of RM07 DNA fragment from Halobacterium halobium in Escherichia coli].

The RM07 DNA fragment from Halobacterium halobium was inserted upstream of chloramphenicol acetyltransferase (cat) reporter gene in pKK232-8 in two different orientations (positively or negatively), generating the RM07-cat fusion plasmids pRM07-1 (+) and pRM07-1 (-). These two plasmids and pKK232-8 were transformed into Escherichia coli HB101 respectively, then the antibiotic resistance level and the chloramphenicol acetyltransferase protein concentration of different transformants were detected. The research results revealed that HB101/pRM07-1 (+) was resistant to chloramphenicol and could grow on the plate containing chloramphenicol, but HB101/pRM07-1 (-) and HB101/pKK232-8 were sensitive to it. Therefore, the results suggested that the positive RM07 fragment had promoter activity in Escherichia coli while the negative RM07 fragment did not. Site-directed mutagenesis of RM07 was performed by PCR mutagenesis method. The effect of specific nucleotide mutations on the chloramphenicol resistance level of different transformants was detected. The different transformants containing different nucleotide mutations were inoculated on the plates containing various concentrations of chloramphenicol and then incubated at the same time. The change of chloramphenicol resistance level reflected the change of promoter activity. By using this method, the important nucleotide responsible for the promoter function of RM07 in Escherichia coli was determined. The promoter activity of RM07 in Escherichia coli was also improved greatly by modifying the nucleotide component.

Chloramphenicol O-Acetyltransferase↗
PubMed2004

[Establishing mouse embryonic stem cell line carrying a fluorescent undifferentiated marker].

To label mouse ES cells,a cell line derived from the inner cell mass of 3.5-day blastocysts,with enhanced green fluorescent protein (EGFP), the vector of pRex-1-EGFP was transferred into mouse ES cells by electroporation. The expressions of Rex-1 in undifferentiated and differentiated ES cells were detected by the microscopic observation of EGFP and by RT-PCR. The results showed that the EGFP gene was transferred into the mouse ES cell line, and the transfected cells in undifferentiated state showed high levels of EGFP expression. When the cells began to differentiate, the EGFP expressions were gradually reduced. A mouse ES cell line expressing EGFP under the control of Rex-1 gene promoter was generated. The cell line provides a powerful approach for the research of the process of mammalian development and for the screening of small molecules that can regulate this process.

Animals↗
PubMed2004

[Combined gene therapy enhances collateral vascularization in coronary artery occlusion swine model].

OBJECTIVE: To investigate the efficacy of combined gene transfer of vascular endothelial growth factor (VEGF) and angiotensin-1 (Ang-1) in swine coronary artery occlusion. METHODS: Swine underwent left thoracotomy followed by ligation of left anterior descending coronary artery. Constructed PCD(2)/VEGF and/or PCD(2)/Ang-1 eukaryotic expression plasmid was directly injected into the swine myocardium. RT-PCR, immunohistochemistry, capillary density and arteriole density were used to detect gene expression and biological effect. Coronary angiography was done to evaluate collateral circulation of the occluded artery. RESULTS: High levels of VEGF and Ang-1 mRNA and protein expression were detected. There was a significant increase in the number of capillaries and arterioles as compared with a control group (P < 0.05). Capillary density and arteriole density of the combination therapy group were higher than those of the swing receiving either therapy alone. Coronary angiography showed better collateral circulation in the combination therapy group. CONCLUSIONS: Direct injection of PCD(2)/VEGF and PCD(2)/Ang-1 can transfect the myocardium and express VEGF and Ang-1 protein. Combined gene transfer of VEGF and Ang-1 can increase capillary and arteriole number and enhance collateral circulation of the occluded coronary artery more effectively.

Angiotensin I↗
PubMed2003

High heparanase activity in multiple myeloma is associated with elevated microvessel density.

Heparanase is an enzyme that cleaves heparan sulfate chains of proteoglycans, and its expression has been associated with increased growth, metastasis, and angiogenesis of some tumors. Because myeloma tumor cells express high levels of the syndecan-1 heparan sulfate proteoglycan and because these tumors grow as highly vascularized aggregates within the bone marrow, we analyzed the activity, expression, and function of heparanase in myeloma patients. Analysis of heparanase activity in the plasma isolated from bone marrow biopsies of 100 patients reveals 86 positive for heparanase activity and 14 negative. The bone marrow samples can be further divided into three categories of heparanase activity, high activity (42 patients), low activity (44 patients), and negative (14 patients). In contrast to the bone marrow plasma, levels of heparanase activity in peripheral blood plasma of 29 myeloma patients were either negative or low, suggesting that in multiple myeloma, heparanase functions in the local microenvironment of the bone marrow and its activity is not significantly elevated systemically. Immunohistochemistry reveals that patients with high levels of heparanase activity often have tumor cells with intense staining for the enzyme. Interestingly, a marked heterogeneity among tumor cells was noted, with clusters of heavily stained cells surrounded by cells with weak or negative staining for heparanase. Analysis of microvessel density reveals a strikingly higher concentration of vessels in patients with high heparanase activity (78.96 vessels/mm(2)) as compared with patients negative for heparanase activity (25.03 vessels/mm(2)). When human myeloma cells transfected with the cDNA for heparanase are implanted in severe combined immunodeficient (SCID) mice, the resulting tumors exhibited a significantly higher microvessel density than did tumors established with control cells. Thus, expression of heparanase appears to play a direct role in enhancing microvessel density in these myeloma tumors. Because heparanase is known to stimulate angiogenesis, and because high microvessel density is associated with poor prognosis in myeloma, we conclude that heparanase expression likely plays an important role in regulating the growth and progression of myeloma, and that therapies designed to block heparanase activity may aid in controlling this cancer.

Animals↗
PubMed2003

[Aprotinin reduces myocardial injury in the off-pump coronary artery bypass grafting].

OBJECTIVE: To investigate the effect of aprotinin on the myocardial injury during the off-pump coronary artery bypass grafting (OPCAB). METHODS: Twenty-four patients undergoing OPCAB randomly were of 12 patients divided into two groups: aprotinin group and Control group (n = 12). In the aprotinin group a loading dose (2 x 10(6) KIU) was administered within the first 30 minutes after the induction followed by continuous infusion of 0.5 x 10(6) KIU/h throughout the operation. The plasma myocardial injury markers CK-MB and cTnI were measured at the four points: (1) After induction; (2) 1 hour after coronary anastomosis; (3) 6 hours and (4) 24 hours after the operation. The blood loss was recorded by autotransfusion in the operation, and the drainage immediately, 6 hours, and 24 hours after the operation. RESULT: Myocardial injury markers CK-MB and cTnI increased significantly after the anastomosis in each group; cTnI was lower in the aprotinin group than in the control group at the third and forth points (P < 0.05). The post-operative drainage after 6 hours and 24 hours were lower in the aprotinin group than in control group (P < 0.05). CONCLUSIONS: There is the myocardial injury during OPCAB. Aprotinin can reduce the myocardial injury during OPCAB.

Aged↗
PubMed2003

Control of NKT cell differentiation by tissue-specific microenvironments.

CD1d-restricted Valpha14 NKT cells play an important role in both Th1- and Th2-type immune responses. To determine whether NKT cells develop two functionally distinct subsets that provoke different types of responses, we examined the phenotypes and cellular functions of NK1.1(+) and DX5(+) T cells. We found that both NK1.1(+) and DX5(+) T cells are CD1d-restricted Valpha14 T cells with identical Ag specificities, phenotypes, tissue locations, and functions. Similar to the NK1.1 marker, the DX5 marker (CD49b) is expressed on mature NKT cells in both NK1.1 allele-positive and allele-negative strains. However, when NK1.1(+) and DX5(+) NKT cells isolated from different tissues were compared, we found that thymic and splenic NKT cells differed not only in their cytokine profiles, but also in their phenotype and requirements for costimulatory signals. Thymic NKT cells displayed the phenotype of activated T cells and could be fully activated by TCR ligation. In contrast, splenic NKT cells displayed the phenotype of memory T cells and required a costimulatory signal for activation. Furthermore, the function and phenotype of thymic and splenic NKT cells were modulated by APCs from various tissues that expressed different levels of costimulatory molecules. Modulation of NKT cell function and differentiation may be mediated by synergic effects of costimulatory molecules on the surface of APCs. The results of the present study suggest that the costimulatory signals of tissue-specific APCs are key factors for NKT cell differentiation, and these signals cannot be replaced by anti-CD28 or anti-CD40 ligand Abs.

Animals↗
PubMed2003

Application of conformation design in acyclic stereoselection: total synthesis of borrelidin as the crystalline benzene solvate.

The total synthesis of (-)-borrelidin (treponemycin), a structurally distinct 18-membered macrolide antibiotic, has been achieved. It was isolated as the crystalline benzene solvate, and its structure was confirmed by a single-crystal X-ray analysis. The deoxypropionate subunit consisting of four alternating C-methyl groups with a C(4)-C(10) syn/syn/anti orientation was elaborated by a new method of iterative cuprate additions to acyclic alpha,beta-unsaturated esters relying on two consecutive 1,3-inductions and starting with d-glyceraldehyde as the chiral progenitor. The unique Z/E cyanodiene unit was obtained as a single isomer by application of the Still-Gennari olefination protocol. The gamma-hydroxycyclopentane carboxylic acid subunit was prepared from L-malic acid utilizing a sequential introduction of C-vinyl and C-allyl groups, capitalizing on 1,2-induction in an acyclic alpha,beta-unsaturated ester and carbocyclization by a Grubbs ring closure metathesis reaction. The prevalence of 1,3-syn-disposed deoxypropionate triads in the cuprate additions is rationalized on the basis of minimized syn-pentane interactions in the transition states. A virtual diamond lattice was used as a visual tool to portray the low-energy conformations of the acyclic substrates, and corroborated by (1)H NMR homodecoupling studies.

Crystallography, X-Ray↗
PubMed2003

Gene and protein expression profiling of human cerebral endothelial cells activated with tumor necrosis factor-alpha.

An increase in permeability of the blood-brain barrier is a critical event in the pathophysiological process of multiple sclerosis and other neurodegenerative diseases. Tumor necrosis factor alpha (TNFalpha) is known to play a crucial role in this process and is a powerful activator of endothelial cell inflammatory responses. Although many reports describe effects of TNFalpha activation in endothelial cells, the molecular mechanisms specific for activation of cerebral endothelial cells remains unclear. The objective of this study was to identify potential pharmaceutical targets for the treatment of multiple sclerosis using molecular profiling techniques. Gene expression measurements (Affymetrix Hu6800 oligonucleotide arrays) and proteomics (two-dimensional gel electrophoresis and mass spectrometry) were applied to analyze early alterations in human cerebral endothelial cells (HCEC) activated by TNFalpha. Human umbilical vein endothelial cells (HUVEC) were used as the reference system. The results presented show that HCEC and HUVEC respond similarly with respect to cell adhesion molecules, chemotaxis, apoptosis and oxidative stress molecules. However, nuclear factors NFkB1 and NFkB2, plasminogen activator inhibitor 1 and cofilin 1 are examples of cerebral specific responses. Our results indicate involvements of the urokinase plasminogen activator system and cytoskeletal rearrangements unique to TNFalpha activation of cerebral endothelial cells.

Antineoplastic Agents↗
PubMed2003

An extensible and systematic force field, ESFF, for molecular modeling of organic, inorganic, and organometallic systems.

ESFF is a rule-based force field designed for modeling organic, inorganic, and organometallic systems. To cover this broad range of molecular systems, ESFF was developed in an extensible and systematic manner. Several unique features were introduced including pseudoangle and a dot product function representing torsion energy terms. The partial atomic charges that are topology-dependent are determined from ab initio (DFT) calculated electronegativity and hardness for valence orbitals. The van der Waals parameters are charge-dependent, and correlated with the ionization potential for atoms in various valence states. To obtain a set of well-defined and physically meaningful parameters, ESFF employs semiempirical rules to translate atomic-based parameters to parameters typically associated with a covalent valence force field. The atomic parameters depend not only on atom type, but also on internal type, thus resulting in a more accurate force field. This article presents the theory and the method used to develop the force field. The force field has been applied to molecular simulations of a wide variety of systems including nucleic acids, peptides, hydrocarbons, porphyrins, transition metal complexes, zeolites, and organometallic compounds. Agreement with the experimental results indicates that ESFF is a valuable tool in molecular simulations for understanding and predicting both crystal and gas phase molecular structures.

Algorithms↗
PubMed2003

Pancreaticobiliary maljunction: pathophysiological and clinical aspects and the impact on biliary carcinogenesis.

BACKGROUND: Pancreaticobiliary maljunction (PBM) is frequently associated with congenital choledochal cyst (CCBD), but differs in embryonic cause and clinical features. It is thought to develop as a misarrangement of the embryonic connections in the pancreaticobiliary ductal system, with the terminal bile duct joined to one of the ducts of the ventral pancreas. Clinical aspects are intermittent abdominal pain, relapsing acute pancreatitis, jaundice, cholangitis, and gallbladder cancer. In patients with PBM and CCBD, primary bile duct stones, acute cholangitis, and bile duct cancer are considered to result from cholestasis, regurgitation of pancreatic juice, and reciprocal reflux of bile and pancreatic juice. The mixture of bile and pancreatic juice due to recipocal reflex very likely plays an important role in biliary carcinogenesis. PATIENTS AND METHODS: We reviewed the pathophysiological and clinical aspects and biliary carcinogenesis in 250 PBM patients (169 with benign hepatobiliary and pancreatic disease, 81 with malignancy). RESULTS: PBM patients show elevated cellular proliferation activity in the gallbladder epithelia. A number of oncogenes and tumor suppressor genes have been identified and implicated in carcinogenesis, particularly the K- ras oncogene and the p53 suppressor gene. Some K- ras mutations do not appear essential for hyperplasia but may be an early event in carcinogenesis. The p53 mutations are involved in carcinogenesis in the biliary epithelium in PBM patients.

Adolescent↗
PubMed2003

Heparan sulfate regulates targeting of syndecan-1 to a functional domain on the cell surface.

In polarized B lymphoid cells, syndecan-1 is targeted specifically to a discrete membrane domain termed the uropod that is located at the cell's trailing edge. Within this functional domain, syndecan-1 promotes cell-cell adhesion and concentration of heparin binding growth factors. The present study reveals the surprising finding that targeting of syndecan-1 to uropods is mediated by its heparan sulfate chains and that targeting is regulated by cell surface events rather than solely by intracellular mechanisms. The addition of exogenous heparin or the treatment of polarized cells with heparitinase initiates a rapid and dramatic redistribution of uropod syndecan-1 over the entire cell surface, and a mutated syndecan-1 lacking heparan sulfate chains fails to concentrate within uropods. Interestingly, the heparan sulfate-bearing proteoglycans glypican-1 and beta glycan fail to concentrate in uropods, indicating that targeting may require heparan sulfate structural motifs unique to syndecan-1 or that the core protein of syndecan-1 participates in specific interactions that promote heparan sulfate-mediated targeting. These findings suggest functional specificity for syndecan-1 within uropods and, in addition, reveal a novel mechanism for the targeting of molecules to discrete membrane subcellular domains via heparan sulfate.

Animals↗
PubMed2003

Dihydropyrimidinase related protein-2 as a biomarker for temperature and time dependent post mortem changes in the mouse brain proteome.

Proteome analysis in the central nervous system area represents a large and important challenge in drug discovery. One major problem is to obtain representative and well characterized tissues of high quality for analysis. We have used brain tissues from normal mice to study the effect of post mortem time (up to 32 h) and temperature (4 degrees C and room temperature) on protein expression patterns. A number of proteins were identified using mass spectrometry and potential markers were localized. One of the proteins identified, dihydropyrimidinase related protein-2 (DRP-2), occurs as multiple spots in two-dimensional electrophoresis gels. The ratio between the truncated form of DRP-2 (fDRP-2) and full length DRP-2 is suggested as an internal control that can be used as a biomarker of post mortem time and post mortem temperature between unrelated brain protein samples. Results of this study may be useful in future efforts to detect disease specific alterations in proteomic studies of human post mortem brain tissues.

Animals↗
PubMed2003

The 492-bp RM07 DNA fragment from the halophilic Archaea confers promoter activity in all three domains of life.

A 492-bp DNA fragment, designated RM07, was isolated from the chromosomal DNA of the halophilic Archaea, Halobacterium halobium, and was shown to confer promoter activity in Escherichia coli. Sequence analysis revealed that RM07 contained three consensus sequences of the archaeal distal promoter element as well as the typical -35 and -10 box sequences of bacterial promoters. Promoter probe analysis confirmed that RM07 conferred promoter activity in all three domains of life: Archaea (Haloferax volcanii), Eukarya (Saccharomyces cerevisiae) and Bacteria (Escherichia coli). Deletion analysis and site-directed mutagenesis further identified the functional regions within RM07 required for promoter activity. This is the first report of a DNA fragment from Archaea that confers promoter activity in all three domains of life, suggesting that the promoter structure and activity may be viewed as a bridge narrowing the gaps among the different domains of life.

Artificial Gene Fusion↗
PubMed2003

Analysis of K-ras codon 12 mutation in flat and nodular variants of serrated adenoma in the colon.

PURPOSE: The developmental process of serrated adenomas is obscure, and the importance of genetic alterations has not been elucidated clearly. The possibility that the developmental process and genetic alterations of serrated adenomas could differ from those of ordinary tubular adenomas was explored in this work. METHODS: Serrated adenomas were obtained by endoscopic resection (n = 57) and divided into two groups: flat (n = 10) and nodular (n = 47). Mutation of the K-ras gene was analyzed by enriched polymerase chain reaction-enzyme-linked mini-sequence assay, which can detect not only the presence of a mutation but also the mutation type of K-ras codon 12 with high sensitivity. Methylation-specific polymerase chain reaction was performed with specific primers for the DNA repair gene O6-methylguanine-DNA methyltransferase. RESULTS: Serrated adenomas located in the rectum were more likely to have a K-ras mutation (9/12, 75 percent), whereas serrated adenomas of the flat type were less likely to have one (1/10, 10 percent). Furthermore, nodular serrated adenomas that occurred in the rectum possessed a high frequency of K-ras gene codon 12 point mutation (8/10, 80 percent) despite an overall frequency of 46.8 percent (22/47). A mutation of the K-ras codon 12 gene was detected in 23 (40.4 percent) of 57 serrated adenomas. Three types of point mutations of codon 12 were detected, with the mutation of GAT being observed most frequently. CONCLUSIONS: This study shows that development of nodular serrated adenomas may depend on the mutation of the K-ras codon 12 gene, whereas development of flat serrated adenomas may not. Additionally, serrated adenomas that occur in the rectum are closely related to the mutation of the K-ras codon 12 gene. K-ras mutations in serrated adenomas may be unaffected by the epigenetic silencing of O6-methylguanine-DNA methyltransferase by promoter hypermethylation.

Adenoma↗
PubMed2003

Rerupture of intracranial aneurysms during cerebral angiography.

Three cases of re-rupture of intracranial aneurysms during cerebral angiography (RIADCA) between June and September, 2001 are reported. All cases underwent emergency craniotomy and aneurysm clipping. The subarachnoid blood and the extravasating contrast medium were removed intraoperatively as completely as possible. There was no mortality in this series. The incidence, timing, sex, age, inducing factors, risk factors, prevention measures and prognosis are discussed and reviewed in conjunction with the literature.

Aged↗