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Biomedical subjects

D Yang

Publications and source records attributed to D Yang.

At least 325 records · Page 18Linked to original sources
PubMed1997

[Genetic divergence of Cordyceps sinensis as estimated by random amplified polymorphic DNA analysis].

Random amplified polymorphic DNA(RAPD) has been used in analyzing 13 winter worm, summer grasses (Cordyceps sinensis [Berk.], Sacc.) from 5 areas in Qingzang plateau. For 19 arbitrary primers, each individual showed about 65 RAPD markers. The genetic distances among those 13 winter worm, summer grasses were correlated to their geographic distances. Molecular phylogenetic tree constructed by UPGMA and NJ suggested that 13 winter worm, summer grasses could be divided into 3 clusters: east cluster, north cluster and middle cluster, in consistence with their origins and genetic divergence among different clusters. Furthermore, our results implied that the RAPD markers of winter worm, summer grasses showed a geographic specificity and could be a better genetic marker in studying genetic diversity and phylogeny of winter worm, summer grasses.

Genetic Variation↗
PubMed1997

Clinical significance of the immunohistochemical detection of p53 protein in endoscopic biopsy specimens.

OBJECTIVE: To evaluate the utility of immunohistochemical staining of p53 expression in endoscopic biopsy specimens in clinical practice. METHODS: Using a streptavidin-peroxidase (S-P) conjugate method with monoclonal p53 antibody Do-7, the expression of p53 was investigated immunohistochemically in 150 endoscopic biopsies and paired brushings from patients with benign and malignant gastric diseases. Among them, 22 cases with gastric carcinoma underwent the gastrectomy and the immunohistochemistry was repeated in the resected cancerous tissues of the same patient. RESULTS: p53 expression in endoscopic specimens was detectable in 35.7% cases of gastric carcinoma (n = 70), 5.9% of gastric polyps (n = 17), 16.7% of gastric mucosal dysplasia (n = 18) and 4.0% of chronic atrophic gastritis (n = 25), but no protein expression was observed in cases of chronic active gastritis (n = 20). In 22 patients with tumor resection, 8 (36.4%) were identified with p53 expression in endoscopic specimens before surgery, while 10 (45.5%) were identified with the same expression in surgical specimens, the real positive rate was 80.0%, no false positives were observed in endoscopic specimens. Furthermore, p53 expression in gastric carcinoma was closely correlated with clinical stage and lymph nodes involvement, but did not correlate with histologic type. CONCLUSIONS: Assay of p53 by immunohistochemistry in endoscopic biopsy specimens and brushings is an easy and reliable technique to assess p53 status in gastric carcinoma, and thus could serve as a marker to foresee the prognosis in these patients and assist in the diagnosis of malignancy before surgery. It may also be a valuable marker in screening patients with high risk of gastric carcinoma.

Adenocarcinoma↗
PubMed1996

Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice.

Programmed cell death (apoptosis) is a prominent feature of the development of the immune and nervous systems. The identification of the Caenorhabditis elegans cell death gene, ced-3, as a prototype of the interleukin-1beta converting enzyme (ICE) protease family has led to extensive evidence implicating these enzymes in apoptosis. Among the ten or more members of the ICE protease family, CPP32/yama/apopain exhibits the highest similarity to CED-3 in both sequence homology and substrate specificity. To analyse its function in vivo, we generated CPP32-deficient mice by homologous recombination. These mice, born at a frequency lower than expected by mendelian genetics, were smaller than their littermates and died at 1-3 weeks of age. Although their thymocytes retained normal susceptibility to various apoptotic stimuli, brain development in CPP32-deficient mice was profoundly affected, and discernible by embryonic day 12, resulting in a variety of hyperplasias and disorganized cell deployment. These supernumerary cells were postmitotic and terminally differentiated by the postnatal stage. Pyknotic clusters at sites of major morphogenetic change during normal brain development were not observed in the mutant embryos, indicating decreased apoptosis in the absence of CPP32. Thus CPP32 is shown to play a critical role during morphogenetic cell death in the mammalian brain.

Animals↗
PubMed1996

Contributions to conformational entropy arising from bond vector fluctuations measured from NMR-derived order parameters: application to protein folding.

The relation between order parameters derived from NMR spin relaxation experiments and the contribution to conformational entropy from ns-ps timescale bond vector dynamics is investigated by considering a number of simple models describing bond vector motion. In a few cases both classical and quantum mechanical derivations are included to establish the validity of obtaining order parameter-entropy relations using classical mechanics only. For these cases it is found that classical and quantum mechanical derivations give very similar results so long as the square of the order parameter of the bond vector is less than approximately 0.95. For a given change in order parameter, the change in conformational entropy is sensitive to the model employed, with the absolute value of the entropy change increasing with the number of degrees of freedom in the model. The entropy-order parameter profile calculated from a 1.12 ns molecular dynamics trajectory of fully hydrated Escherichia coli ribonuclease HI is well fit using a simple expression based on a model assuming bond vector diffusion in a cone, suggesting that it may well be possible to extract meaningful entropy changes reflecting changes in ps-ns time scale motions from changes in NMR-derived order parameters. Contributions to the conformational entropy change associated with a folding-unfolding transition of an SH3 domain and calculated from changes in rapid N-HN backbone dynamics are presented.

Entropy↗
PubMed1996

Human hepatic lipase subunit structure determination.

Chinese hamster ovary cells were stably transfected with a human hepatic lipase (HL) cDNA. The recombinant enzyme was purified from culture medium in milligram quantities and shown to have a molecular weight, specific activity, and heparin affinity equivalent to HL present in human post-heparin plasma. The techniques of intensity light scattering, sedimentation equilibrium, and radiation inactivation were employed to assess the subunit structure of HL. For intensity light scattering, purified enzyme was subjected to size exclusion chromatography coupled to three detectors in series: an ultraviolet absorbance monitor, a differential refractometer, and a light scattering photometer. The polypeptide molecular weight (without carbohydrate contributions) was calculated using the measurements from the three detectors combined with the extinction coefficient of human HL. A single protein peak containing HL activity was identified and calculated to have a molecular mass of 107,000 in excellent agreement with the expected value for a dimer of HL (106.8 kDa). In addition, sedimentation equilibrium studies revealed that HL had a molecular mass (with carbohydrate contributions) of 121 kDa. Finally, to determine the smallest structural unit required for lipolytic activity, HL was subjected to radiation inactivation. Purified HL was exposed to various doses of high energy electrons at -135 degrees C; lipase activity decreased as a single exponential function of the radiation dose to less than 0.01% remaining activity. The target size of functional HL was calculated to be 109 kDa, whereas the size of the structural unit was determined to be 63 kDa. These data indicate that two HL monomer subunits are required for lipolytic activity, consistent with an HL homodimer. A model for active dimeric hepatic lipase is presented with implications for physiological function.

Animals↗
PubMed1996

Malignancy in chronic leg ulcers.

OBJECTIVE: To evaluate the frequency of malignant ulcers in patients presenting with leg ulcers. DESIGN: A descriptive study from data collected between July 1988 and June 1995 from 981 patients (2448 ulcers) attending a leg ulcer clinic. SETTING: A specialised leg ulcer clinic at a tertiary teaching hospital. SUBJECTS: 43 patients with 55 malignant skin lesions. OUTCOME MEASURES: Tissue biopsies in ulcerated lesions that suggested malignancy or were not responding to appropriate treatment. RESULTS: Forty-three patients were found to have malignant lesions on the legs, giving a frequency of malignant ulcers of 4.4 per 100 leg ulcer patients, or 2.2 per 100 leg ulcers. Seventy-five per cent of the malignant ulcers were basal cell carcinoma and 25% were squamous cell carcinoma. CONCLUSIONS: Malignant skin changes are common in chronic leg ulcers. A biopsy should be taken from all suspicious ulcers or ulcers that do not respond to appropriate treatment.

Aged↗
PubMed1996

Hepatic overexpression of insulin-like growth factor-II in adulthood increases basal and insulin-stimulated glucose disposal in conscious mice.

The physiological role of circulating insulin-like growth factor-II (IGF-II) in adult humans is poorly understood. We recently generated an IGF-II transgenic murine model of persistent IGF-II production (plasma IGF-II approximately 30-fold increased above normal) through over-expression of the transgene driven by the major urinary protein promoter (Rinderknecht, E., and Humbel, R. E. (1978) J. Biol. Chem. 269, 13779-13784). To determine whether in vivo insulin action is improved in these transgenic mice, we performed euglycemic insulin (18 milliunits/kg.min) clamp studies in conscious IGF-II transgenic and in age- and weight-matched control mice. Plasma glucose and insulin concentrations were significantly lower in the IGF-II transgenic compared with both control grouoff Despite decreased plasma glucose concentration, basal hepatic glucose production (HGP) and glucose clearance were increased. During the insulin clamp studies in IGF-II transgenic mice compared with control mice (a) the rates of glucose infusion and glucose uptake were increased by approximately by 65 and approximately 55%, respectively; (b) glycolysis was increased by approximately 12% while glycogen synthesis was approximately 2-fold higher; (c) while the suppression of plasma free fatty acid was similar, the increment in plasma lactate concentration was significantly higher; (d) although HGP was similarly inhibited by insulin, phosphoenolpyruvate gluconeogenesis was enhanced and accounted for a larger portion of HGP (64% versus approximately 40% in control mice). Our data suggest that the persistence of circulating IGF-II in adult mice to levels commonly observed in adult humans (50-70 nM) causes a marked improvement in peripheral (skeletal muscle) insulin action, which is not due to changes in body composition. These results suggest that circulating IGF-II may exert a regulatory role on insulin sensitivity and body composition in humans.

Adult↗
PubMed1996

Characterization of an artificial dimer of ribonuclease H using 1H NMR spectroscopy.

The protein fusion technique was applied in the synthesis of an artificial dimer of ribonuclease H (305 residues). 1H NMR spectroscopy was used to analyze the structure of this dimer. Spectral profiles and pKa values of the histidine residues obtained using 1H NMR indicate that the dimer retains the secondary and tertiary structures of the intact monomer. Selective spin-lattice relaxation measurements suggest that the two monomeric units in the dimer are in tight contact. Furthermore, the 2D 1H NMR and paramagnetic relaxation filter results show that the two monomers bind together through interactions between the N- and C-terminal sites of the linked regions.

Amino Acid Sequence↗
PubMed1996

The significance of heregulin in breast cancer tumor progression and drug resistance.

The erbB-2 receptor plays an important role in the prognosis of breast cancer and is expressed at high levels in nearly 30% of tumors in breast cancer patients. While evidence accumulates to support the relationship between erbB-2 overexpression and poor overall survival in human breast cancer, understanding of the biological consequence(s) of erbB-2 overexpression remains elusive. The discovery of heregulin has allowed us to identify a number of related but distinct biological endpoints which appear responsive to signal transduction through the erbB-2/4 receptor. These endpoints of growth, invasiveness, and differentiation have clear implications for the emergence, maintenance, and/or control of malignancy, and represent established endpoints in the assessment of malignant progression in human breast cancer. Preliminary studies in vitro have shown that heregulin induces a biphasic growth effect on cells with erbB-2 overexpression. Interestingly, we observed that expression of heregulin correlates with a more aggressive/invasive, vimentin-positive phenotype in breast cancer cells lines. Therefore, we have postulated that heregulin is involved in breast cancer tumor progression. We have shown that heregulin induces in vitro chemoinvasion and chemotaxis of breast cancer cells as well as growth in an anchorage dependent and independent manner. Interestingly, a heregulin neutralizing antibody inhibits chemotaxis and results in cell growth inhibition and blockade of the invasive phenotype. Strikingly, genetically engineered cells which constitutively express heregulin demonstrate critical phenotypic changes that are associated with a more aggressive phenotype. Specifically, these cells are no longer dependent on estrogen for growth and are resistant to tamoxifen in vitro and in vivo, and moreover these cells metastasize to lymph nodes in athymic nude mice. These tumors appear to have lost bcl-2 expression as compared with the control tumors. In addition, presumably by activation/regulation of topoisomerase II, the heregulin-transfected cells become exquisitely sensitive to doxorubicin and VP-16. Clearly, mechanistic aspects of the erbB-2/4 and heregulin interaction need to be understood from a therapeutic standpoint which could provide additional insights into synergistic treatments for certain patients, or improve treatment regimens for a large number of women. The study of heregulin and its co-expression with erbB-2/4 receptor and the assessment of its involvement in the progression from the in situ stage of breast tumors to the invasive one will additionally increase the relevance of heregulin as a prognostic/diagnostic factor. We believe that our studies provide new insights into breast cancer diagnosis, prognosis, and treatment.

Breast Neoplasms↗
PubMed1996

Intracytoplasmic sperm injection improving embryo quality: comparison of the sibling oocytes of non-male-factor couples.

OBJECTIVE: Our objective was to investigate whether the quality of embryos developed after intracytoplasmic sperm injection (ICSI) is better than that of conventional IVF embryos. METHODS: Nine couples who previously achieved a normal rate of fertilization following IVF and four couples whose normal rate of fertilization was expected were involved in this study. The oocytes from those couples were randomly divided into two groups, group A by conventional insemination and group B by ICSI. The fertilization rate and quality of embryos were compared. RESULTS: Normal fertilization was achieved in 61% of the oocytes (83/136) after conventional insemination. In group B, 69% of the oocytes (99/144) achieved normal fertilization, although only 127 metaphase II oocytes were injected using the ICSI technique. More grade A embryos were obtained when the ICSI technique was used for fertilization than by conventional IVF (35.4 and 24.3%, respectively; P = 0.028). CONCLUSIONS: A similar fertilization rate can be achieved by ICSI in comparison with conventional IVF, when male factor is not involved. Embryos after ICSI have an improved quality.

Adult↗
PubMed1996

[Detection and significance of HCV RNA in saliva, seminal fluid and vaginal discharge in patients with hepatitis C].

To investigate the transmission of HCV infection through family contact, we detected HCV RNS in body fluids (saliva, seminal and vaginal discharge) of 16 serum HCV RNA positive patients (including 7 men and 9 women) and in sera of their family members. The positive rates of HCV RNA in the body fluids were 31.25% (5/16) in saliva, 57.14% (4/17) in seminal fluid and 22.22% (2/9) in vaginal discharge, respectively. Among the family members in our series, all were negative for both anti-HCV and HCV RNA, despite two spouses positive for HCV RNA. This result strongly suggested the potential possibility of the transmission of HCV infection through the family contact.

Adult↗
PubMed1996

The synthesis of fluorine-18 lomefloxacin and its preliminary use in human studies.

Lomefloxacin is a new fluorine-containing antibiotic that has recently been approved for general use. Fluorine-18 lomefloxacin has been prepared by fluoride exchange between fluorine-18 fluoride and lomefloxacin in DMSO. Both time and temperature of the reaction have been optimized and conditions developed for the isolation and purification of the labeled product in a form suitable for oral administration. The exchange reaction provides sufficient labeled material for human studies with pharmacologically relevant quantities of the drug. We have performed preliminary human studies with this compound using positron emission tomography to estimate the tissue distribution of the compound and show the distribution of the compound into the liver and lungs.

Animals↗
PubMed1996

Elevation of nitrate levels in pregnant ewes and their fetuses.

OBJECTIVE: Nitric oxide is a potent vasodilator released by endothelial cells that may play an important role in modulating maternal and fetal vascular tone in normal pregnancy. The current study was designed to evaluate whether plasma or urine nitrite and nitrate (the metabolites of nitric oxide) concentrations are elevated in pregnant compared with those of nonpregnant sheep and whether the nitrate concentrations in the fetal circulation were increased in comparison with the maternal circulation. STUDY DESIGN: Eleven pregnant sheep and seven nonpregnant oophorectomized sheep were instrumented with catheters in the maternal and fetal femoral arteries and veins, uterine and umbilical veins, and amniotic cavity. Blood, urine, and amniotic fluid samples were collected for nitrate determination at least 5 days after surgery. After extraction nitrate was reduced to nitrite and quantitated with the Greiss reagent. RESULTS: Arterial plasma nitrate concentrations in the pregnant sheep were significantly elevated compared with those of nonpregnant sheet (5.0 +/- 0.9 vs 2.5 +/- 0.6 micromol/L, p < 0.05). The urinary nitrate concentrations were also significantly increased in the pregnant sheep compared with those of nonpregnant sheet (89.9 +/- 16.3 vs 23.1 +/- 4.5 nmol/mg creatinine, p < 0.01). Fetal plasma nitrate concentrations were ninefold higher than the maternal nitrate concentrations (43.9 +/- 7 vs 5.0 +/- 0.9 micromol/L, p < 0.01), whereas amniotic fluid concentrations were extremely high (133.8 +/- 13.8 micromol/L, n = 3). No venous-arterial differences were measurable across either the maternal or fetal sides of the placenta. CONCLUSION: Nitrate concentrations in pregnant sheet and their fetuses are increased. The increased nitrate concentrations in the maternal and fetal circulations may reflect the increased nitric oxide synthesis, which may in part mediate the cardiovascular adaptations to normal pregnancy and the low systemic and umbilical vascular resistance in the fetus.

Amniotic Fluid↗
PubMed1996

Insulin and angiotensin II are additive in stimulating TGF-beta 1 and matrix mRNAs in mesangial cells.

Angiotensin II (Ang II) and insulin are implicated in the mesangial cell hypertrophy and excessive accumulation of mesangial matrix seen in glomerulosclerosis. Therefore, the effects of Ang II with and without insulin on mRNA levels of several important extracellular matrix genes and transforming growth factor beta-1 (TGF-beta 1) were examined. Ang II alone (1 microM) added to quiescent, murine mesangial cells in serum-free, insulin-free media slightly but not significantly increased TGF-beta 1, fibronectin, collagen I, collagen IV and laminin message levels. The slight elevations in message expression were reversed by losartan, suggesting that these modest effects are mediated by the AT-1 receptor. Ang II alone also had no significant effects on TGF-beta 1 and extracellular matrix message levels in quiescent rat mesangial cells. In contrast, significant increases in mRNA for collagen 1 (6-fold), collagen IV (4-fold), fibronectin 1 (4-fold) and TGF-beta 1 (2-fold) were seen with insulin alone (10(-6)M) in rat mesangial cells, and a dose-response effect could be demonstrated for insulin (10(-9) to 10(-6)M). Ang II plus insulin further significantly increased collagen I (9-fold), collagen IV (9-fold), fibronectin 1 (5-fold) and TGF-beta 1 (3-fold) message expression. These effects were partially reversed in the presence of losartan. The Northern analyses were supported by measurements of active and total TGF-beta 1 activity (pg/ml/ 5 x 10(6) cells): 1145 +/- 76 and 1960 +/- 199, serum free control; 1121 +/- 92 and 1932 +/- 214, Ang II (10(-6)M); 4589 +/- 103 (P < 0.001 vs. control) and 11071 +/- 1952 (P < 0.01 vs. control), insulin (10(-6)M); and 6881 +/- 183 (P < 0.001 vs. control) and 16626 +/- 1435 (P < 0.01 vs. control), insulin plus Ang II. These results suggest that insulin, itself, significantly increases TGF-beta 1 and extracellular matrix gene expression in rat mesangial cells. Ang II alone has modest effects, while Ang II and insulin have additive effects. To explain the mechanism of these additive effects, we investigated the action of Ang II on insulin signaling and the effect of insulin on Ang II AT1 receptor mRNA expression. Ang II did not enhance insulin-induced insulin receptor substrate-1 (IRS-1) phosporylation or phosphatidylinositol3 (PI-3) kinase activity, but did enhance insulin-induced mitogen activated protein (MAP) kinase activity. Insulin increased message levels of AT1 receptor by twofold. These results suggest that enhancement of MAP kinase activity and AT1 receptor regulation by insulin may contribute to the additive effects of insulin and Ang II in mesangial cells.

Angiotensin II↗
PubMed1996

Structural effect of intra-strand cisplatin-crosslink on palindromic DNA sequences.

Three self-complementary DNA oligonucleotides, each having a single GpG site, have been reacted with the anticancer platinum compound cis-Pt(NH3)2Cl2 (cisplatin). Their covalent intra-strand didentate adducts have been purified and studied by NMR. In d(GAC-CATATG*G*TC), the two G*G* sites (G*G* denoting the cisplatin crosslinked lesion site) are separated by four base pairs and capped by two base pairs at the ends of the helix and the dodecamer forms a doubly-kinked duplex structure. Multi-stranded aggregate of the dodecamer was formed over time in the presence of chloride. This is due to the meta-stable property of the intra-strand Pt-G*pG* crosslink in dsDNA, similar to that first seen recently in another duplex (Yang et al., Biochemistry (1995) 34, 12912-12920). In d([c7A]CC[c7G][c7G]CCG*G*T), the CG*G*T segment of the decamer is essentially single-stranded with the G*8 in the syn conformation. In d([c7G]CC[c7G]CG*G*C), two possible structures, a full duplex and a staggered partial duplex, were formed. Therefore, the structural consequence of the incorporation of the G*G* lesion site into palindromic sequences is dependent on the location of the lesion sites in the sequence. The destabilizing effect of G*G* in dsDNA may facilitate the formation of a hairpin structure as shown recently (Iwamoto et al., J. Amer. Chem. Soc. (1994) 116, 6238-6244). Such alternative structural distortions may be relevant in understanding the protein recognition of the lesions induced by cisplatin.

Binding Sites↗
PubMed1996

Localization and clinical significance of thyrotropin receptor mRNA expression in orbital fat and eye muscle tissues from patients with thyroid-associated ophthalmopathy.

We have studied the cellular localization of thyrotropin receptor (TSH-R) mRNA in orbital fat and extraocular muscle tissues from patients with thyroid-associated ophthalmopathy (TAO) using Northern blot, reverse transcriptase polymerase chain reaction (RT-PCR), and in situ hybridization, and we correlated the findings with clinical estimates of ophthalmopathy. Although we failed to detect TSH-R mRNA in orbital tissues by Northern blot, TSH-R cDNA was amplified in orbital fat tissue from 13 of 25 patients with TAO and from 2 of 4 control subjects, in eye muscle tissue from 2 out of 7 patients with TAO, and in cultured orbital fibroblasts and subcutaneous fibroblasts from TAO patients. In situ hybridization showed that TSH-R mRNA was detected in cultured orbital fibroblasts as well as skin fibroblasts obtained from the patient. Furthermore, the expression of TSH-R mRNA in orbital fat tissue from patients with TAO significantly correlated with the orbital fat volume and the severity of ophthalmopathy, especially the extent of eye muscle dysfunction. These results suggest that the expression of TSH-R in the orbit, especially fibroblasts, may play a role in the pathogenesis and clinical manifestations of the ophthalmopathy in patients with TAO, although a secondary effect, involving fibroblasts in TAO is also possible.

Adipose Tissue↗