Search PubMed⌕ Search

Biomedical subjects

Y Tang

Publications and source records attributed to Y Tang.

At least 37 records · Page 2Linked to original sources
PubMed2001

Increased inwardly rectifying potassium currents in HEK-293 cells expressing murine transient receptor potential 4.

Drosophila transient receptor potential (Trp) and its mammalian homologues are postulated to form capacitative Ca2+ entry or store-operated channels. Here we show that expression of murine Trp4 in HEK 293 cells also leads to an increase in inwardly rectifying K+ currents. No similar increase was found in cell lines expressing Trp1, Trp3 or Trp6. Consistent with typical characteristics of inward rectifiers, the K+ currents in Trp4-expressing cells were blocked by low millimolar concentrations of Cs+ and Ba2+, but not by 1.2 mM Ca2+, and were only slightly inhibited by 5 mM tetraethylammonium. Single channel recordings of excised inside-out patches revealed the presence of two conducting states of 51 pS and 94 pS in Trp4-expressing cells. The outward current in the excised patches was blocked by 1 mM spermine, but not by 1 mM Mg2+. How Trp4 expression causes the increase in the K+ currents is not known. We propose that Trp4 either participates in the formation of a novel K+ channel or up-regulates the expression or activity of endogenous inwardly rectifying K+ channels.

Animals↗
PubMed2001

X-Ray study on an artificial mung bean inhibitor complex with bovine beta-trypsin in neat cyclohexane.

The active trypsin inhibiting component, SPC1, was obtained during the synthesis of a 22-residue peptide with three disulfide bridges according to the mimic mung bean Bowman-Birk type inhibitor. The K(i) value of SPC1 is 1.2x10(-7) M. In order to determine the topological structure of SPC1, X-ray diffraction studies were carried out on the complex of SPC1 with bovine beta-trypsin. Only the binding loop of SPC1 resolved at 2.2 A resolution due to conformational flexibility of the other residues [1]. The amino acid sequence was re-determined and electrospray mass spectroscopy was also performed to ensure that no cleaving occurred on SPC1 and the primary sequence of SPC1 is correct. Because the protein is more rigid in nonaqueous medium as has been proved by others [2], we treated the complex of SPC1 with neat cyclohexane and then subjected it to X-ray diffraction analysis, and the result showed that all the 22 residues of SPC1 were located in the electron density map. So the topological structure of SPC1 has been determined, suggesting that crystal treatment with cyclohexane may be used as a method to determine the conformation of the disordered regions in protein crystal structures.

Amino Acid Sequence↗
PubMed2001

Stabilization of coiled-coil peptide domains by introduction of trifluoroleucine.

Substitution of leucine residues by 5,5,5-trifluoroleucine at the d-positions of the leucine zipper peptide GCN4-p1d increases the thermal stability of the coiled-coil structure. The midpoint thermal unfolding temperature of the fluorinated peptide is elevated by 13 degrees C at 30 microM peptide concentration. The modified peptide is more resistant to chaotropic denaturants, and the free energy of folding of the fluorinated peptide is 0.5-1.2 kcal/mol larger than that of the hydrogenated form. A similarly fluorinated form of the DNA-binding peptide GCN4-bZip binds to target DNA sequences with affinity and specificity identical to those of the hydrogenated form, while demonstrating enhanced thermal stability. Molecular dynamics simulation on the fluorinated GCN4-p1d peptide using the Surface Generalized Born implicit solvation model revealed that the coiled-coil binding energy is 55% more favorable upon fluorination. These results suggest that fluorination of hydrophobic substructures in peptides and proteins may provide new means of increasing protein stability, enhancing protein assembly, and strengthening receptor-ligand interactions.

Amino Acid Sequence↗
PubMed2001

[Analysis of 2200 kidney transplantations].

OBJECTIVE: To analyze the clinical results of 2 200 renal transplantations. METHODS: The following factors were analyzed: (1) the graft survival rates at 1, 3 and 5 years; (2) main factors contributing to the mortality; (3) factors affecting the survival rate of retransplant grafts; (4) features of cancers after renal transplantation; (5) effect of CSA on liver function after renal transplantation; (6) treatments for chronic rejection; and (7) effect of HLA typing on renal transplantation. RESULTS: From 1972 to 2000, 2 200 renal transplantations were performed in 1 908 patients (1 337 men and 571 women) with end-stage renal failure in our hospital, Beijing. The graft survival rates at 1, 3 and 5 years were 87.3%, 80.2% and 67.0% respectively, after using CSA since 1985. 302 recipients were aged over 50 years whose one year graft survival rate was 83.4% and one year patient survival rate was 85.4%. Pneumonitis was the dominant death factor after renal transplantation, with an incidence of 4.49% and a death rate of 34.62%. The incidence of positive CMV after renal transplantation was up to 40.3%, in with a death rate of 8.0%. The incidence of tumor after renal transplantation was 1.5%. The most frequent site of tumor was urological system. The patient's liver function before transplantation and the intraindividual variability of bioavailabity to CSA etc. were the major toxic factors of the liver. The incidences of acute rejection were 27%, 35.5%, 22.4%, 20.85%, 0% and 0%, respectively, when the HLA mismatches were 6, 5, 4, 3, 2 and 1. CONCLUSION: Using immunosuppressents reasonably, preventing acute rejection, and achieving good HLA typing are the major factors improving long-term graft/patient survival rate.

Adolescent↗
PubMed2001

Substantial differences in specificity of HIV-specific cytotoxic T cells in acute and chronic HIV infection.

Cytotoxic T lymphocytes (CTLs) play a vital part in controlling viral replication during human viral infections. Most studies in human infections have focused on CTL specificities in chronic infection and few data exist regarding the specificity of the initial CTL response induced in acute infection. In this study, HIV-1 infection in persons expressing human histocompatibility leukocyte antigen (HLA)-A*0201 was used as a means of addressing this issue. In chronic infection, the dominant HLA-A*0201-restricted CTL response is directed towards the epitope SLYNTVATL ("SL9") in p17 Gag (residues 77-85). This epitope is targeted by 75% of HLA-A*0201-positive adults, and the magnitude of this A*0201-SL9 response shows a strong negative association with viral load in progressive infection. Despite using the highly sensitive peptide-major histocompatibility complex tetramer and intracellular cytokine assays, responses to the SL9 epitope were not detectable in any of 11 HLA-A*0201-positive subjects with acute HIV-1 infection (P = 2 x 10(-6)), even when assays were repeated using the SL9 peptide variant that was encoded by their autologous virus. In contrast, multiple responses (median 3) to other epitopes were evident in 7 of the 11 A*0201-positive subjects. Longitudinal study of two subjects confirmed that the A*0201-SL9 response emerged later than other CTL responses, and after viral set point had been reached. Together, these data show that the CTL responses that are present and that even may dominate in chronic infection may differ substantially from those that constitute the initial antiviral CTL response. This finding is an important consideration in vaccine design and in the evaluation of vaccine candidates.

Acute Disease↗
PubMed2001

Blood genomic responses differ after stroke, seizures, hypoglycemia, and hypoxia: blood genomic fingerprints of disease.

Using microarray technology, we investigated whether the gene expression profile in white blood cells could be used as a fingerprint of different disease states. Adult rats were subjected to ischemic strokes, hemorrhagic strokes, sham surgeries, kainate-induced seizures, hypoxia, or insulin-induced hypoglycemia, and compared with controls. The white blood cell RNA expression patterns were assessed 24 hours later using oligonucleotide microarrays. Results showed that many genes were upregulated or downregulated at least twofold in white blood cells after each experimental condition. Blood genomic response patterns were different for each condition. These results demonstrate the potential of blood gene expression profiling for diagnostic, mechanistic, and therapeutic assessment of a wide variety of disease states.

Animals↗
PubMed2001

Hierarchical approach to predicting permeation in ion channels.

A hierarchical computational strategy combining molecular modeling, electrostatics calculations, molecular dynamics, and Brownian dynamics simulations is developed and implemented to compute electrophysiologically measurable properties of the KcsA potassium channel. Models for a series of channels with different pore sizes are developed from the known x-ray structure, using insights into the gating conformational changes as suggested by a variety of published experiments. Information on the pH dependence of the channel gating is incorporated into the calculation of potential profiles for K(+) ions inside the channel, which are then combined with K(+) ion mobilities inside the channel, as computed by molecular dynamics simulations, to provide inputs into Brownian dynamics simulations for computing ion fluxes. The open model structure has a conductance of approximately 110 pS under symmetric 250 mM K(+) conditions, in reasonable agreement with experiments for the largest conducting substate. The dimensions of this channel are consistent with electrophysiologically determined size dependence of quaternary ammonium ion blocking from the intracellular end of this channel as well as with direct structural evidence that tetrabutylammonium ions can enter into the interior cavity of the channel. Realistic values of Ussing flux ratio exponents, distribution of ions within the channel, and shapes of the current-voltage and current-concentration curves are obtained. The Brownian dynamics calculations suggest passage of ions through the selectivity filter proceeds by a "knock-off" mechanism involving three ions, as has been previously inferred from functional and structural studies of barium ion blocking. These results suggest that the present calculations capture the essential nature of K(+) ion permeation in the KcsA channel and provide a proof-of-concept for the integrated microscopic/mesoscopic multitiered approach for predicting ion channel function from structure, which can be applied to other channel structures.

Bacterial Proteins↗
PubMed2001

Coumaroyl flavonol glycosides from the leaves of Ginkgo biloba.

Two coumaroyl flavonol glycosides, isorhamnetin 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside], and kaempferol 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside]-7-O-beta-D-glucopyranoside, were isolated from the n-BuOH extract of Ginkgo biloba leaves. These two, together with six other flavonol glycosides, kaempferol 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside], quercetin 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside], quercetin 3-O-alpha-L-[6"'-p-coumaroyl-(beta-D)-glucopyranosyl-(1,2)-rhamnopyranoside]-7-O-beta-D-glucopyranoside, quercetin 3-O-beta-D-glucopyranosyl-(1-2)-alpha-L-rhamnopyranoside, quercetin 3-O-beta-rutinoside, and quercetin 3-O-beta-D-glucopyranoside, showed profound antioxidant activities in DPPH and cytochrome-c reduction assays using the HL-60 cell culture system.

Antioxidants↗
PubMed2001

Phosphodiesterase 3B gene expression is enhanced in the liver but reduced in the adipose tissue of obese insulin resistant db/db mouse.

Phosphodiesterase (PDE) 3B, when activated by insulin, causes a decrease in intracellular cAMP concentration. The activation of this enzyme results in the reduced output of free fatty acids (FFA) from adipocytes, and an increased lipogenesis in liver. We have recently shown that PDE3B gene expression is reduced in adipose tissues of KKAy mice. We intend to further elucidate the regulation of PDE3B in liver as well as adipose tissues in relation to the insulin resistant state. We examined PDE3B gene expression in liver and adipose tissues of obese, insulin-resistant diabetic db/db mice and also checked the effect of an insulin-sensitizing drug, troglitazone, on this gene expression. In the liver of db/db mice, PDE3B mRNA, its corresponding protein, and the associated catalytic activity were all increased by 2.1, 1.9 and 1.6-fold, respectively, over those in db/+ control mice. Histological examination revealed substantial triglyceride storage in the liver of db/db mice. Conversely, in the adipose tissue of db/db mice, PDE3B mRNA, protein, and its associated activity were all decreased by 0.38, 0.33 and 0.36-fold, respectively. Troglitazone, which has no effect on PDE3B in liver, increased the expression of this gene in adipocytes. This increase is associated with a reduction in the elevated levels of serum insulin, glucose, FFA and triglycerides. The reduced PDE3B gene expression in adipose tissues, which results in the elevation of serum FFA, could be the primary event in the development of insulin resistance in db/db mice. The enhanced PDE3B gene expression may correlate with changes in triglyceride storage in the liver of these mice.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed2001

Oestrogen-mediated suppression of tumour necrosis factor alpha-induced apoptosis in MCF-7 cells: subversion of Bcl-2 by anti-oestrogens.

In oestrogen receptor (ER)-positive breast carcinoma cells, 17beta-oestradiol suppresses a dose-dependent induction of cell death by tumour necrosis factor alpha (TNF). The ability of oestrogens to promote cell survival in ER-positive breast carcinoma cells is linked to a coordinate increase in Bcl-2 expression, an effect that is blocked with the pure anti-oestrogen ICI 182,780. The role of Bcl-2 in MCF-7 cell survival was confirmed by stable overexpression of Bcl-2 which resulted in suppression of apoptosis induced by doxorubicin (DOX), paclitaxel (TAX) and TNF as compared to vector-control cells. The pure anti-oestrogen ICI 182,780 in combination with TNF, DOX or TAX potentiated apoptosis in vector-transfected cells. Interestingly, pre-treatment with ICI 182,780 markedly enhanced chemotherapeutic drug- or TNF-induced apoptosis in Bcl-2 expressing cells, an effect that was correlated with ICI 182,780 induced activation of c-Jun N-terminal kinase. Our results suggest that the effects of oestrogens/anti-oestrogens on the regulation of apoptosis may involve coordinate activation of signalling events and Bcl-2 expression.

Antibiotics, Antineoplastic↗
PubMed2001

Four new isoflavone triglycosides from Sophora japonica.

Four new isoflavone triglycosides, genistein 7-O-beta-D-glucopyranoside-4'-O-[(alpha-L-rhamnopyranosyl)-(1-->2)-beta-D-glucopyranoside] (1), genistein 7-O-beta-D-glucopyranoside-4'-O-[(beta-D-glucopyranosyl)-(1-->2)-beta-D-glucopyranoside] (2), genistein 7-O-alpha-L-rhamnopyranoside-4'-O-[(alpha-L-rhamnopyranosyl)-(1-->2)-beta-D-glucopyranoside] (3), and genistein 7-O-alpha-L-rhamnopyranoside-4'-O-[(beta-D-glucopyranosyl)-(1-->2)-beta-D-glucopyranoside] (4), together with nine known compounds, namely, genistein 7-O-beta-D-glucopyranoside-4'-O-beta-D-glucopyranoside, sophorabioside, prunetin 4'-O-beta-D-glucopyranoside, sophororicoside, genistin, rutin, kaempferol 3-O-beta-rutinoside, quercetin 3-O-beta-D-glucopyranoside, and kaempferol 3-O-beta-D-glucopyranoside, were isolated from the pericarps of Sophora japonica. The structures of 1-4 were determined by spectroscopic methods.

Carbohydrate Sequence↗
PubMed2001

Crystal structure of the complex of concanavalin A and tripeptide.

The X-ray structure analysis of a cross-linked crystal of concanavalin A soaked with the tripeptide molecule as the probe molecule showed electron density corresponding to full occupation in the binding pocket. The site lies on the surface of concanavalin A and is surrounded by three symmetry-related molecules. The crystal structure of the tripeptide complex was refined at 2.4-A resolution to an R-factor of 17.5%, (Rfree factor of 23.7%), with an RMS deviation in bond distances of 0.01 A. The model includes all 237 residue of concanavalin A, 1 manganese ion, 1 calcium ion, 161 water molecules, 1 glutaraldehyde molecule, and 1 tripeptide molecule. This X-ray structure analysis also provides an approach to mapping the binding surface of crystalline protein with a probe molecule that is dissolved in a mixture of organic solvent with water or in neat organic solvent but is hardly dissolved in aqueous solution.

Binding Sites↗
PubMed2001

Crystal structure of the complex of concanavalin A and hexapeptide.

The X-ray structure analysis of a cross-linked crystal of concanavalin A soaked with a hexapeptide molecule as a probe molecule showed an electron density corresponding to full occupation in the binding pocket. The site lies on the surface of concanavalin A and is surrounded by three symmetry-related molecules. The crystal structure of the hexapeptide complex was refined at 1.93-A resolution, to an R-factor of 19% (Rfree factor of 25%), with an RMS deviation in bond distances of 0.01 A. The model includes all 237 residues of concanavalin A, one manganese ion, one calcium ion, 95 water molecules, one glutaraldehyde molecule, one isopropanol molecule, and one hexapeptide molecule. This X-ray structure analysis also provides an approach to mapping the binding surface of crystalline protein with a probe molecule that is dissolved in the mixture of organic solvent with water or in neat organic solvent but is hardly dissolved in aqueous solution.

Binding Sites↗
PubMed2001

A mimotope of pre-S2 region of surface antigen of viral hepatitis B screened by phage display.

To acquire the phage-displayed mimotopes which mimic the specificity of hepatitis B virus surface antigen (HBsAg), a random peptide library expressing linear peptide with 12 amino acids in length were used to screen with the serum from a hepatitis B virus infected patient in the recovery phase. After 3 rounds of biopanning, the positive phages were confirmed by competitive ELISA using HBsAg/P33. Two phagotopes were identified and one of them was confirmed as mimotope by competition experiment. Based on the mimotpe, a multiple antigenic peptide with four branches was synthesized by solid phase peptide synthesis. The antiginicity and specificity of the synthesized antigen was tested in BALB/c mice compared with the native epitope-based antigen. The results showed that the mimotope-based antigen could evoke higher titer of antibodies with the same specificity of the epitope-based antigen. Those findings indicate mimotopes can be used in antigen and vaccine design.

Animals↗
PubMed2001

The biomechanical, morphologic, and histochemical properties of the costal cartilages in children with pectus excavatum.

BACKGROUND/PURPOSE: The cause of the pectus excavatum (PE) remains unclear, although some results of research have indicated that the disturbance of the sternum or costal cartilage might be responsible for this deformity. But no decisive evidence has been gained. The authors have analyzed the biomechanical, morphologic, and histochemical properties of the cartilage in PE and intend to support the belief that the disturbance of the cartilage might contribute to the development of PE. METHODS: Thirty-eight specimens of the sixth cartilage were obtained at operation for the PE group (aged from 3 to 6 years; mean, 4.2 years). And 28 specimens of the control group (aged from 3 to 6 years; mean, 4.4 years) were gained from routine postmortem examinations in which the cause of death was unlikely to have affected the cartilage. The biomechanical test was carried out in a material testing machine (Shimadzu AG-10TA, Tokyo, Japan). The relation curve of load-deformation in tensile and compressive tests and the curve of load-time in the flexuous test were recorded automatically. The values of the ultimate strength and strain were calculated from this relation curve. The specimens also underwent H&E staining. The values of the area, circumference, mean diameter, maximal diameter, and morphologic factor of the cell and the nucleus of the cartilage in superficial and deep area were determined with the help of image analysis software (GT-2 model, China). The superficial zone (SZ) and deep zone (DZ) of the cartilage were examinated with electron microscopy (JEM-100SX, Japan). The distribution and intensity of type II collagen was shown by immunohistochemistry staining and analyzed with the image analysis software (GT-2 model, Huakang Co, Chengdu, China). The extent and distribution of proteoglycan were analyzed after Safranin-O and periodic acid shiff (PAS) staining. RESULTS: The mean strength of the costal cartilage in the experimental group was less than that in the control group in terms of tension, compression, and flexure (P <.05). The shape of the stress-strain curve for tension and compression in the experimental group was different from the control group. The fracture load in the experimental group was less than in the control group in tension (1.5 MPa versus 2.8 MPa) and in compression (.2 MPa versus 8.3 MPa). The time of fracture in experimental group was 30 seconds compared with 38 seconds in control group. No denaturation or necrosis could be found in light microscopical examination. There was no manifestation of hyperplasia or hypoplasia in the costal cartilage of the PE group. In SZ and DZ areas, the pattern and the number of mitochondria, endoplasmic reticulum, and Golgi in the experimental group were the same as the control group in transmission electron microscopy. Furthermore, the distribution and the number of proteoglycan in the 2 groups did not show a significant difference both in SZ and DZ areas. Although the distribution of the collagen in SZ areas was normal, this pattern was disturbed in DZ areas in the experiment group. The results of type II collagen immunohistochemistry examination was concordant with that change. No significant difference between control and experimental group could be seen in Safranin-O and PAS staining for proteoglycan. CONCLUSIONS: The biomechanical stability of the cartilage was decreased in the PE group. This might be caused by the disorderly arrangement and distribution of the collagen in the cartilage of PE patients. J Pediatr Surg 36:1770-1776.

Biomechanical Phenomena↗
PubMed2001

In vitro affinity maturation of human IgM antibodies reactive with tumor-associated antigens.

Human lymphocytes secreting tumor cell-specific IgM antibodies were enriched in vitro following the stimulation of allogeneic human splenocytes from nontumor-bearing donors with cytostatic tumor cells or tumor cell plasma membrane fractions. The antibodies were generally of the IgM class and displayed low intrinsic affinity (K(d) > 100 nM). Nonetheless, the avidity arising from multivalent binding sites permitted the identification of multiple monoclonal antibodies (MAbs) displaying specificity for cultured tumor cells. Five antibodies were cloned from the B cells and two of these were expressed as human Fabs with IgG(1) constant regions. Although the avidity of the human IgM antibodies was sufficient to permit detection in the original screening, the monovalent Fabs displayed low binding activities, consistent with their low intrinsic affinity. Thus, in vitro affinity maturation was used to rapidly generate multiple variants of both antibodies displaying greater than 100-fold higher affinity. Two of the antibodies were characterized further and shown to have distinct specificities. One of the targets, LH11238, is associated both with the plasma membrane and with lysosomes and is rapidly internalized following incubation of the antibody with intact live cell monolayers. The second antigen, designated LH13, is a secreted antigen that has been enriched 200-fold from conditioned media and consists of two reactive bands at 42 and 45 kDa on denaturing Western blots. The stimulation and enrichment of human lymphocytes in culture coupled with rapid in vitro affinity maturation of low affinity antibodies potentially enables the discovery of human antibodies to a broader range of epitopes, including those that might be of greater therapeutic relevance.

Amino Acid Sequence↗