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Biomedical subjects

S Yu

Publications and source records attributed to S Yu.

At least 163 records · Page 9Linked to original sources

[Study on method of sampling and analysis of acephate in the air].

Acephate in the air was collected with polyurethane foamed plastics, desorbed with methanl, separated with a column 2% DEGS and determined by GC-FPD. The detectable limit was 0.052 mg/m3. When the concentration of standard solution was 3-15 micrograms/ml, the relative standard deviation(RSD) was 4.5%-2.3%. The desorption efficiency was above 85%. The sampling efficiency was 100%. The samples were stable for 24 h. It was proved that the method was accurate, fast and simple. It was suitable for the determination of acephate in the air.

Air Pollutants, Occupational↗

[The changes of GFAP and PCNA after human brain contusion: an immunohistochemical study in forensic pathology].

In order to find out changes of GFAP and PCNA after human brain trauma, samples of 38 cases were studied by the immunnohistochemistry method combined with quantitative image analysis. The gray degree and area of PCNA positive cells increased to the maximum at the 4th day and then decreased gradually. GFAP positive cells began to increase significantly 12 hours after contusion, reached the maximum at the 4th day and stayed in high level until the 7th day. These results suggest that: (1) GFAP, PCNA show regular changes with survival time after brain contusion; (2) GFAP and PCNA can be used as time markers of brain contusion from 2-20 days.

Adult↗

[Fibronectin in adhesion, spreading and proliferation of mandibular condylar cartilage cells on cytodex-3 microcarrier].

OBJECTIVE: The purpose of this study was to investigate the effects of exogenous fibronectin on the attachment and growth of mandibular condylar cartilage (MCC) cells on DEAE-dextran micro-carrier. METHODS: MCC cells were harvested from newborn New Zealand white rabbits by sequential digestion with trypsin and collagenase. They were grown on the 20 mg/L fibronectin-coated and uncoated cytodex-3 micro-carrier. Samples were collected after 1, 3, 5, 7, 9, 11 days. The kinetics of adhesion and growth were observed with phase contrast microscope and environmental scanning electronic microscope (ESEM) and quantified by 0.1% crystal violet nuclear extrusion method. RESULTS: MCC cells could rapidly attach and spread onto the fibronectin-coated cytodex-3, and nearly 60 percent cells attached to the micro-carrier within 2 hours. Spreading cells were flat and lowly refractile observed under phase contrast microscope. While the attachment and spreading of the cells in the untreated group were slow, and only 36 percent cells attached to the uncoated micro-carrier. However, most cells attached to the micro-carrier in both groups after 24 hours. The MCC cells attaching to FN-coated cytodex-3 observed under ESEM were flatter and adhered tightly with more processes (pseudopodia) stretching out from cytoplasm. In contrast, cells on untreated cytodex-3 remained regularly round even 24 hours after being plated. Meanwhile, there was an accelerated growth rate of the MCC cells on the fibronectin treated cytodex-3. The cell density in the FN treated group was higher than that of the untreated group at the end of the culture. CONCLUSION: Fibronectin, one major glycoprotein of extracellular matrix, plays an important role in cell attachment and spreading. The better understanding of the extracellular matrix will be helpful in selecting the optimal substrate for cell growth in vitro.

Adhesiveness↗

[The immunohistochemical and electron microscopical study of pathological classification of Melkersson-Rosenthal syndrome].

OBJECTIVE: To prove further the pathological classification of Melkersson-Rosenthal syndrome (MRS) and observe the morphological features of the granuloma cells in MRS. METHODS: We studied 20 cases of MRS, with 10 cases of granuloma type and the other 10 cases of non-granuloma type. The negative results of periodic acid-schiff (PAS), Neelren anti-acid special staining and tuberculosis (TB)-DNA polymerase chain reaction proved that all cases were not TB and fungous disease. Then all the cases were labelled with antibody of lysozyme by immunohistochemical staining and investigated with electron microscope. We studied the expression of lysozyme in multinucleated giant cells, epithelioid cells and histiocytes. RESULTS: The results of lysozyme staining were positive in the multinucleated giant cells, epithelioid cells and histiocytes of the granuloma type and were negative in those cells of the non-granuloma type. On the other hand, the small multinucleated giant cells which had 2-3 nuclei were the features of the morphological structure of the granuloma type. The small multinucleated giant cells were not seen in the non-granuloma type. CONCLUSION: In this paper, two pathological types (granuloma and non-granuloma type) in the MRS were proved further and it is found that the different structures of granuloma can be used to discriminate between MRS and sarcoidosis and tuberculosis.

Diagnosis, Differential↗

[Effects of cultivating measures on root rot of Panax notoginseng].

OBJECTIVE: To probe into the effects of cultivating measures on root rot of Panax notoginseng for the integrate control. METHOD: Field investigation on the planting plots was conducted. CONCLUSIONS: Effects of cultural system on root rot are obvious, (1) The rotation period should be more than eight years; (2) Improper fertilizer application is one important fact of the root rot occuring; (3) The light transmitting rate is one important fact; (4) Laying film and using sun-shade shed can reduce the root rot.

Fertilizers↗

[Transgenic mice model of human hepatitis B virus x gene].

OBJECTIVE: To set up a human hepatitis B virus x (HBx) gene transgenic model. METHODS: The model was set up by microinjection, the integration and expression of the founders were identified by molecular hybridization. RESULTS: A 1,800 bp DNA fragment containing the human HBx promoter and genomic DNA was microinjected into the male pronucleus of the fertilized eggs of mice. Three hundred and thirty-eight eggs survived from microinjection were transplanted into the oviduct of 16 pseudopregnant female, 12 pseudopregnant mice got pregnant and gave birth to 74 pups. Genomic DNA were extracted from the tails of the pups, Southern blot hybridization was used for determination of the integration of human HBx gene. The 1,800 bp microinjected fragment was used as the probe, 17 mice (founders) were identified of the integration of human HBx gene. Total RNA were extracted from the livers of the founders, all 17 founders were identified of the expression of human HBx gene by using Northern blot hybridization. CONCLUSIONS: These results may provide animal models for studying on the correlation between chronic HBV infection and the development of hepatocellular carcinoma.

Animals↗

Regulation of BAD phosphorylation at serine 112 by the Ras-mitogen-activated protein kinase pathway.

The function of the pro-apoptotic molecule BAD is regulated by phosphorylation of two sites, serine-112 (Ser-112) and serine-136 (Ser-136). Phosphorylation at either site results in loss of the ability of BAD to heterodimerize with the survival proteins BCL-XL or BCL-2. Phosphorylated BAD binds to 14-3-3 and is sequestered in the cytoplasm. It has been shown that phosphorylation of BAD at Ser-136 is mediated by the serine/threonine protein kinase Akt-1/PKB which is downstream of phosphatidylinositol 3-kinase (PI3K). The signaling process leading to phophorylation of BAD at Ser-112 has not been identified. In this study, we show that phosphorylation of the two serine residues of BAD is differentially regulated. While Ser-136 phosphorylation is concordant with activation of Akt, Ser-112 phosphorylation does not correlate with Akt activation. Instead, we demonstrate that activated Ras and Raf, which are upstream of mitogen-activated protein kinases (MAPK), stimulate selective phosphorylation of BAD at Ser-112. Furthermore, phosphorylation of Ser-112, but not Ser-136 requires activation of the MAPK pathway as the MEK inhibitor, PD 98059, blocks EGF-, as well as activated Ras- or Raf-mediated phosphorylation of BAD at Ser-112. Therefore, the PI3K-Akt and Ras-MAPK pathways converge at BAD by mediating phosphorylation of distinct serine residues.

3T3 Cells↗

alpha-1,4-glucan lyases producing 1,5-anhydro-D-fructose from starch and glycogen have sequence similarity to alpha-glucosidases.

In the past few years a novel enzyme alpha-1,4-glucan lyase (EC 4.2. 2.13), which releases 1,5-anhydrofructose from starch and glycogen, has been cloned and characterized from red algae and fungi. Accumulated evidence indicates that the lytic degradation of starch and glycogen also occurs in other organisms. The present review focuses on the biochemical and molecular aspects of eight known alpha-1,4-glucan lyases and their genes from red algae and fungi. While the amino acid sequence identity is 75-80% among the alpha-1, 4-glucan lyases from each of the taxonomic groups, the identity between the algal and fungal alpha-1,4-glucan lyases is only 25-28%. Notably database searches disclosed that the alpha-1,4-glucan lyases have a clear identity of 23-28% with alpha-glucosidases of glycoside hydrolase family 31, thus for the first time linking enzymes from the class of hydrolases with that of lyases. The alignment of lyases and alpha-glucosidases revealed seven well-conserved regions, three of which have been reported to be involved in catalysis and substrate binding in alpha-glucosidases. The shared substrate and inhibitor specificity and sequence similarity of alpha-1,4-glucan lyases with alpha-glucosidases suggest that related structural elements are involved in the two different catalytic mechanisms.

Amino Acid Sequence↗

Differential targeting of protein kinase CK2 to the nuclear matrix upon transient overexpression of its subunits.

Modest dysregulation of CK2 has been shown to enhance the oncogenic potential in transgenic models of cancer. Since nuclear matrix serves as an anchor for CK2 and plays a key role in growth-related activities, we examined the effects of CK2 overexpression on its signaling to the nuclear matrix. Expression plasmids pCI-CK2alpha, pCI-CK2beta, and the bicistronic pCI-CK2alphabeta containing full length cDNAs encoding the various subunits were employed to transiently transfect two cell lines, BPH-1 and COS-1. Cytosol from transfected BPH-1 cells containing alpha or beta or alpha + beta or alphabeta showed a modest increase in CK2 activity by 26%, 1%, 20%, and 17%, respectively, over that in the controls transfected with pCI vector. However, the corresponding increase in CK2 activity in the NM fraction was 156%, 8%, 147%, and 152%, respectively. Immunoblot analysis of the CK2 in the NM accorded with these data. Similar results were obtained with COS-1 cells or other expression vectors. The results suggest that moderate overexpression of CK2 in the cells evokes a differential several-fold enhancement in NM associated CK2 relative to that in the cytosol. This process may have a bearing on the functional signaling of this kinase in relation to its possible role in oncogenesis.

Androgen-Binding Protein↗

A group of alpha-1,4-glucan lyases and their genes from the red alga Gracilariopsis lemaneiformis: purification, cloning, and heterologous expression.

We present here the first report of a group of alpha-1,4-glucan lyases (EC 4.2.2.13) and their genes. The lyases produce 1, 5-anhydro-D-fructose from starch and related oligomers and polymers. The enzymes were isolated from the red alga Gracilariopsis lemaneiformis from the Pacific coasts of China and USA, and the Atlantic Coast of Venezuela. Three lyase isozymes (GLq1, GLq2 and GLq3) from the Chinese subspecies, two lyase isozymes (GLs1 and GLs2) from the USA subspecies and one lyase (GLa1) from the Venezuelan subspecies were identified and investigated. GLq1, GLq3, GLs1 and GLa1 were purified and partially sequenced. Based on the amino acid sequences obtained, three lyase genes or their cDNAs (GLq1, GLq2 and GLs1) were cloned and completely sequenced and two other genes (GLq3 and GLs2) were partially sequenced. The coding sequences of the lyase genes GLq1, GLq2 and GLs1 are 3267, 3276 and 3279 bp, encoding lyases of 1088, 1091 and 1092 amino acids, respectively. The deduced molecular masses of the mature lyases from the coding sequences are 117030, 117667 and 117790 Da, respectively, close to those determined by mass spectrometry using purified lyases. The amino acid sequence identity is more than 70% among the six algal lyase isozymes. The algal GLq1 gene was expressed in Pichia pastoris and Aspergillus niger, and the expression product was identical to the wild-type enzyme.

Amino Acid Sequence↗

Multiple oligodeoxyribonucleotide syntheseson a reusable solid-phase CPG support via the hydroquinone-O,O'-diacetic acid (Q-Linker) linker arm.

A strategy for oligodeoxyribonucleotide synthesis on a reusable CPG solid-phase support, derivatized with hydroxyl groups instead of amino groups, has been developed. Ester linkages, through a base labile hydroquinone- O, O '-diacetic acid ( Q-Linker ) linker arm, were used to couple the first nucleoside to the hydroxyl groups on the support. This coupling was rapidly accomplished (10 min) using O -benzotriazol-1-yl- N, N, N ', N '-tetramethyluronium hexafluorophosphate (HBTU) and 1-hydroxybenzotriazole as the activating reagents. Oligodeoxyribonucleotide synthesis was performed using existing procedures and reagents, except a more labile capping reagent, such as chloro-acetic anhydride, methoxyacetic anhydride or t-butylphenoxyacetic anhydride, was used instead of acetic anhydride. After each oligodeoxyribonucleotide synthesis, the product was cleaved from the support with ammonium hydroxide (3 min) and deprotected as usual. Residual linker arms or capping groups were removed by treatment with ammonium hydroxide/methylamine reagent and the regenerated support was capable of reuse. Up to six different oligodeoxyribonucleotide syntheses or up to 25 cycles of nucleoside derivatization and cleavage were consecutively performed on the reusable support. This method may provide a significant cost advantage over conventional single-use solid supports currently used for the manufacture of antisense oligodeoxyribonucleotides.

Glass↗

Nuclear matrix targeting of the protein kinase CK2 signal as a common downstream response to androgen or growth factor stimulation of prostate cancer cells.

Protein kinase CK2, a messenger-independent serine/threonine kinase, has been implicated in cell growth. Androgenic stimulus in rat prostate modulates its association with nuclear matrix (NM) and chromatin. Because the growth of human prostate carcinoma cells is influenced by androgens and/or growth factors, we determined the nature of CK2 signaling in the NM in response to androgen and growth factor stimuli. Androgen-sensitive LNCaP and androgen-insensitive PC-3 cells were cultured in media to regulate their growth in the presence of 5alpha-dihydrotestosterone (5alpha-DHT) or growth factors (epidermal growth factor, keratinocyte growth factor, and transforming growth factor alpha). The activity of CK2 was measured in the cytosolic and NM fractions isolated from these cells after treatment with growth stimuli. The changes in CK2 in various fractions were also confirmed by immunoblotting with a specific antibody. LNCaP cells responded to both 5alpha-DHT and growth factors for growth. The presence of these agents in the culture medium evoked a translocation of CK2 to the NM from the cytosol. The PC-3 cells did not respond to 5alpha-DHT for growth but did respond to growth factors. Under these conditions, there was also a translocation of CK2 to the NM concomitant with a decrease in the cytosolic fraction. These results suggest that CK2 translocation to the NM occurs in response to various growth stimuli in cells in culture. Thus, CK2 is a common downstream signal transducer in response to diverse growth stimuli that may relate to the pathobiology of prostate cancer cells.

Adenocarcinoma↗

Mutagenesis of the conserved residue Glu259 of Gsalpha demonstrates the importance of interactions between switches 2 and 3 for activation.

We previously reported that substitution of Arg258 within the switch 3 region of Gsalpha impaired activation and increased basal GDP release due to loss of an interaction between the helical and GTPase domains (Warner, D. R., Weng, G., Yu, S., Matalon, R., and Weinstein, L. S. (1998) J Biol. Chem. 273, 23976-23983). The adjacent residue (Glu259) is strictly conserved in G protein alpha-subunits and is predicted to be important in activation. To determine the importance of Glu259, this residue was mutated to Ala (Gsalpha-E259A), Gln (Gsalpha-E259Q), Asp (Gsalpha-E259D), or Val (Gsalpha-E259V), and the properties of in vitro translation products were examined. The Gsalpha-E259V was studied because this mutation was identified in a patient with Albright hereditary osteodystrophy. S49 cyc reconstitution assays demonstrated that Gsalpha-E259D stimulated adenylyl cyclase normally in the presence of GTPgammaS but was less efficient with isoproterenol or AlF4-. The other mutants had more severely impaired effector activation, particularly in response to AlF4-. In trypsin protection assays, GTPgammaS was a more effective activator than AlF4- for all mutants, with Gsalpha-E259D being the least severely impaired. For Gsalpha-E259D, the AlF4--induced activation defect was more pronounced at low Mg2+ concentrations. Gsalpha-E259D and Gsalpha-E259A purified from Escherichia coli had normal rates of GDP release (as assessed by the rate GTPgammaS binding). However, for both mutants, the ability of AlF4- to decrease the rate of GTPgammaS binding was impaired, suggesting that they bound AlF4- more poorly. GTPgammaS bound to purified Gsalpha-E259D irreversibly in the presence of 1 mM free Mg2+, but dissociated readily at micromolar concentrations. Sucrose density gradient analysis of in vitro translates demonstrated that all mutants except Gsalpha-E259V bind to beta gamma at 0 degreesC and were stable at higher temperatures. In the active conformation Glu259 interacts with conserved residues in the switch 2 region that are important in maintaining both the active state and AlF4- in the guanine nucleotide binding pocket. Although both Gsalpha Arg258 and Glu259 are critical for activation, the mechanisms by which these residues affect Gsalpha protein activation are distinct.

Aluminum Compounds↗

Selective expression of folate receptor beta and its possible role in methotrexate transport in synovial macrophages from patients with rheumatoid arthritis.

OBJECTIVE: To investigate the expression of folate receptors (FR) and reduced folate carrier (RFC) and determine their relevance to methotrexate (MTX) transport in synovial mononuclear cells (SMC) from patients with rheumatoid arthritis (RA). METHODS: Levels of FR and RFC messenger RNA (mRNA) were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) in SMC from RA patients and peripheral blood mononuclear cells from healthy donors. Expression of FR-beta mRNA and protein was determined by Northern blot and Western blot analyses in RA SMC and monocyte/macrophage-lineage cells. FR-beta expression and folic acid binding capacity on the cell surface were examined by flow cytometric analysis and 3H-folic acid binding analysis. Studies of the inhibition of 3H-MTX uptake in the presence of unlabeled folic acid were performed to investigate the uptake of MTX through FR in RA SMC. RESULTS: RT-PCR, Northern blot, and Western blot analyses showed that FR-beta mRNA and protein were expressed selectively in activated monocytes and CD14+ RA SMC. These cells exhibited folic acid binding capacity. Furthermore, the FR-beta protein was shown to have folic acid binding capacity. Uptake of 3H-MTX through RA SMC was significantly inhibited in the presence of unlabeled folic acid. CONCLUSION: These results demonstrate that FR-beta expression is selectively elevated in RA synovial macrophages and suggest that MTX is transported through FR-beta in RA synovial macrophages. The findings suggest that folate antagonists with higher affinity for FR-beta would be useful in the treatment of RA.

Arthritis, Rheumatoid↗

Cigarette smoking among medical college students in Wuhan, People's Republic of China.

BACKGROUND: This study examined cigarette smoking among a sample of medical college students in Wuhan, People's Republic of China. Specific cultural factors associated with cigarette smoking in China were discussed and issues which need to be studied in the future are presented. METHODS: A self-administrated questionnaire was administered to a sample of 1,611 medical students in 1995. Smoking status was based on self-reported use of cigarettes within the preceding 180 days. Information about demographic characteristics, home residence, annual family income, and smoking status of family members was also obtained. RESULTS: A total of 1,540 (1,201 males and 339 females) completed and returned the questionnaires (response rate 95.6%). None of the 339 female students surveyed reported cigarette smoking. A total of 37.7% (95% confidence interval (CI) = 35.0-40.5%) of the male medical students were current smokers, and 5. 3% (95% CI = 4.0-6.6%) were former smokers. The mean age of beginning to smoke was 16.5 years (95% CI = 8.7-24.3 years). The major reasons for first smoking were stress (42.8%, 95% CI = 38.2-47. 4%), curiosity (34.4%, 95% CI = 29.6-38.4%), and loneliness (33.7%, 95% CI = 28.7-37.4%). Multivariate analyses suggested that age, college year, and having a family member who smoked were significantly associated with cigarette smoking. CONCLUSIONS: The results indicated a need to provide comprehensive tobacco use prevention programs among medical students.

Adolescent↗

Endothelial function and carotid artery wall thickening in patients with early essential hypertension.

To investigate the relationship between endothelial function and carotid artery wall thickening in patients with early mild essential hypertension, the percent dilatation of humerus diameter and intimal-medial thickening (IMT) of common carotid artery was measured by using high-resolution ultrasound in 20 patients with early mild essential hypertension and 18 patients with normotension. The patients with hypertension had not been treated and their history of increased blood pressure was less than 12 months. In essential hypertension group, the percent dilatation of humerus diameter decreased significantly (2.65 +/- 0.98% vs 6.38 +/- 1.61%); IMT of carotid artery increased (0.88 +/- 0.16 mm vs 0.58 +/- 0.08). There was significant negative correlation between IMT and the percent dilatation of humerus diameter (gamma = -0.82, P < 0.05), and no correlation between IMT and 24 h mean systolic, diastolic pressure (gamma = 0.12 and gamma = 0.07, respectively; P > 0.05). Our results suggested that there was endothelial dysfunction in early mild essential hypertension. Endothelial dysfunction may not only contribute to the pathogenesis of hypertension but also serve as the most important inducing factor leading transformation from hypertension to atherosclerosis.

Atherosclerosis↗

Phospholipase D as an effector for ADP-ribosylation factor in the regulation of vesicular traffic.

A mammalian phospholipase D (PLD) activity that is stimulated by ADP-ribosylation factor (ARF) has been identified in Golgi-enriched membrane fractions. This activity is due to the PLD1 isoform and evidence from several laboratories indicates that PLD1 is important for the polymerization of vesicle coat proteins on membranes. When expressed in Chinese hamster ovary cells, PLD1 localized to dispersed small vesicles that overlapped with the location of the ERGIC53 protein, a marker for the endoplasmic reticulum (ER)-Golgi intermediate compartment. Cells having increased PLD1 expression had accelerated anterograde and retrograde transport between the ER and Golgi. Membranes from cells having elevated PLD1 activity bound more COPI, ARF, and ARF-GTPase activating protein. These membranes also produced more COPI vesicles than did membranes from control cells. It is likely that PLD1 participates in both positive and negative feedback regulation of the formation of COPI vesicles and is important for controlling the rate of this process.

ADP-Ribosylation Factors↗