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Biomedical subjects

Y Xue

Publications and source records attributed to Y Xue.

At least 217 records · Page 12Linked to original sources

Serum and tissue levels of estradiol during estrogen-induced renal tumorigenesis in the Syrian hamster.

The estrogen-induced renal tumor in the hamster has emerged as a major animal model in hormonal carcinogenesis. However, a fundamental aspect of this experimental model has as yet not been investigated. In the present study, comparisons between the serum and tissue 17 beta-estradiol (E2) levels in cyclic female hamsters and corresponding hormone levels in E2-treated castrated male hamsters have been made. Data is provided concerning the concentration of estrogenic hormones in the serum and target tissue typically required to elicit renal tumorigenesis in this species. Serum E2 levels in the cyclic female hamster average 79 pg/ml on days 1-2 and 311 pg/ml on days 3-4, attaining a maximum of 358 pg/ml on day 4 of the cycle. Elevation in uterine, renal and hepatic E2 tissue levels during days 3-4 of the cycle reflect increases in serum E2 levels which were 3.0-, 2.0-, and 2.6-fold higher when compared to day 1 of the cycle in these tissues. As expected, serum E2 levels of untreated castrated male hamsters did not appreciably vary over a 6 month period of aging and averaged about 32 pg/ml. Under conditions which produced essentially 100% renal tumor incidence, a rapid rise in serum E2 levels, averaging 71.0-fold higher than untreated castrated levels, was seen. A steady state serum E2 level of 2400 to 2700 pg/ml was maintained from 45-180 days of continuous estrogen treatment. Compared to kidneys of untreated hamsters, renal E2 levels in E2-treated hamsters rose only on average 5.4-fold between 15-180 days of hormone exposure. Serum levels of E2-treated hamsters were 5.7- to 8.0-fold higher than those observed in cyclic female hamsters on days 3 and 4. However, at these higher E2-treated serum levels there was no apparent effect either on weight loss or mortality of the animals.

Animals↗

Cloning and structural analysis of the anthocyanin pigmentation locus Rt of Petunia hybrida: characterization of insertion sequences in two mutant alleles.

Anthocyanin biosynthesis in flowers of Petunia hybrida is controlled by the regulatory genes an1, an2 and an11. Seven classes of cDNA clones homologous to transcripts that are down-regulated in an1-, an2- and an11- mutants were isolated via differential cDNA cloning. Genetic mapping, antisense RNA experiments and analyses of mutant alleles demonstrated that one class of clones originated from the Rt locus. The rt gene has no introns and encodes a protein with homology to mammalian glucuronosyl transferases and flavonoid 3-O-glucosyltransferase (UF3GT) encoded by the bz1 gene from Zea mays. As the Rt locus controls the rhamnosylation of reddish anthocyanin-3-O-glucosides which is the first in a series of modifications that finally yield magenta or blue/purple coloured anthocyanins, this suggests that rt encodes an anthocyanin rhamnosyl transferase. Molecular analysis of two mutant rt alleles showed that their expression is blocked by different DNA insertion elements. Mutability of the rt-vu15 allele results from the presence of a 284 bp transposable element (dTph1) in the rt promoter region, causing a block in transcription. The protein coding region of the rt-r27 allele contains a 442 bp insertion (dTph3) resulting in premature polyadenylation of rt transcripts. Although dTph3 cannot transpose, it has sequence characteristics of transposable elements, suggesting that it is a defective member of a new family of transposable elements.

Alleles↗

Mutations in the Caenorhabditis elegans beta-tubulin gene mec-7: effects on microtubule assembly and stability and on tubulin autoregulation.

We have sequenced 45 mutations in mec-7, a beta-tubulin gene required for the production of 15-protofilament microtubules in the nematode Caenorhabditis elegans, and have correlated sequence alterations with mutant phenotypes. The expression patterns of most alleles have also been determined by in situ hybridization and immunocytochemistry. Most (12/16) complete loss-of-function alleles, which are recessive, result from nonsense mutations, insertions, or deletions; three others disrupt a putative GTP-binding domain. Three of the four loss-of-function, missense mutations result in elevated mec-7 message levels, suggesting a defect in tubulin autoregulation that may be attributable to a loss in the ability to form heterodimers. Most (8/9) mild alleles are caused by missense mutations. Two mild alleles appear to increase microtubule stability and lead to the elaboration of ectopic neuronal processes in mec-7-expressing cells. Most (15/23) mutations that cause severe dominant or semidominant phenotypes are clustered into three discrete domains; four others occur in putative GTP-binding regions. Many of these dominant mutations appear to completely disrupt microtubule assembly.

Alleles↗

Effects of liuwei dihuang wan [symbol: see text:bd and some other TCM drugs on bone biomechanics and serum 25 (OH)D3 content in rats.

After a relatively long-term injection of hydrocortisone into rats, the strength of bone (anti-stress capacity) reduced evidently, while the rigidity (anti-deformity capacity) increased markedly (ie, bone fragility elevated). At the same time, the content of serum 25(OH)D3 decreased remarkably. After oral administration of Liuwei Dihuang Wan ([see symbol: see text]), anti-stress capability of bone increased evidently and its anti-deformity capability returned to normal; however, there was no elevation of serum 25(OH)D3 content. The effect of Longmu Zhuanggu Chongji ([see symbol: see text]) or Jisheng Shenqi Wan (see symbol: see text]) was lower than that of Liuwei Dihuang Wan. The experiments suggests that Liuwei Dihuang Wan is beneficial to preventing and curing osteoporosis, but no correlation between its mechanism and the metabolism of 25(OH)D3 was demonstrated.

Animals↗

[PCR detecting homologous recombinants from ES cells].

PCR is an effective method for detecting homologous recombinants from ES cells, because there are different genomic DNA structure among non-recombinants, site specific recombinants and non-site specific recombinants. When we design suitable primers, we can analyze and identify site specific recombinants, non-site specific recombinants and non-recombinants. In this paper, we design two different pairs of primer (according to the genomic DNA patterns) for using PCR to detect the cells which resist to G418 and 6-TG after plasmid pRV4.0 transforming male mouse ES cells, we get the homologous recombinants quickly, simply and specifically. We test the reliability of PCR using southern blot hybridization.

Animals↗

[Studies on deoxynivalenol contamination in grain and its products in Anhui Province].

Contamination of deoxynivalenol (DON) in wheat, corn, barley, and wheat flour produced in Anhui Province were examined with thin-layer chromatography in 1983-1991. In total, 672 specimens were determined, and 63.2% of them were contaminated by DON, with an average contents of 908.7 micrograms/kg (a range of 0-13330 micrograms/kg). The contamination of DON in wheat and wheat flour was more severe than that in corn (P < 0.01). There was highly significant difference in the contamination of DON in wheat produced in different areas and years, with a P-value less than 0.01 by ANOVA, but no significant difference in corn (P > 0.1). It suggested the contamination of DON correlated with the incidence of scabby wheat in that area during these years.

Food Contamination↗

Cloning and expression of cDNA encoding Xenopus laevis bone morphogenetic protein-1 during early embryonic development.

The Xenopus laevis DNA fragment encoding a protein homologous with human bone morphogenetic protein-1 (BMP-1) was amplified by polymerase chain reaction (PCR) from cDNA derived from stage 26 (st.26) embryonic RNA. Subsequently this fragment was used as a probe to isolate cDNA clones by screening of a X. laevis st.24 embryonic cDNA library. Two partial clones (22 and 63) were obtained and the missing 5'-end of the clone 22 was extended by the anchored PCR technique. The nucleotide sequence of the resulting clone (22AN) contained an open reading frame coding for a protein with 707 deduced amino acids. Three sizes of mRNA (2.9, 5.2 and 6.6 kb) were detected in blastula (st.9) and early gastrula (st.10) embryos, and in hatched tadpole (st.40), but little or no expression was observed in morula (st.7) and late gastrula (st.12) embryos, suggesting a physiological role(s) of X.laevis BMP-1 in normal embryonic development.

Amino Acid Sequence↗

Importance of the conserved active-site residues Tyr7, Glu106 and Thr199 for the catalytic function of human carbonic anhydrase II.

The catalytic mechanism of carbonic anhydrase includes the reaction of a zinc-bound hydroxide ion with the CO2 substrate. This hydroxide ion is part of a hydrogen-bonded network involving the conserved amino acid residues Thr199, Glu106 and Tyr7. To investigate the functional importance of these residues, a number of site-specific mutants have been made. Thus, Thr199 has been changed to Ala, Glu106 to Ala, Gln and Asp, and Tyr7 to Phe. The effects of these mutations on catalyzed CO2 hydration and ester hydrolysis have been measured, as well as the binding of some inhibitors. The results show that the CO2 hydration activity of the mutant with Phe7 is only marginally reduced, whereas the esterase activity is larger than that of unmodified enzyme. It is concluded that Tyr7 is not a functionally required element of the hydrogen-bonded network. The CO2 hydration activity (kcat as well as kcat/Km) and the esterase activity of the mutant with Ala199 are reduced about 100-fold. The affinity for the sulfonamide inhibitor, dansylamide, is only slightly reduced while the mutant has an enhanced affinity for bicarbonate and the anionic inhibitor, SCN-. The activities of the mutants with Ala106 and Gln106 are also reduced. The reduction of the esterase activity is about 100-fold, while kcat for CO2 hydration has decreased by a factor of 1000. The parameter kcat/Km is only about one order of magnitude smaller for these mutants than for the unmodified enzyme. The binding of dansylamide and another sulfonamide inhibitor, acetazolamide, are about 20-times weaker to the mutant with Gln106 than to unmodified enzyme. These results suggest important roles for Thr199 and Glu106 in carbonic anhydrase catalysis. The mutant with Asp106 is almost fully active suggesting that the structure has undergone a compensatory change to maintain the interaction between residue 106 and Thr199.

Amino Acid Sequence↗

Crystallographic analysis of Thr-200-->His human carbonic anhydrase II and its complex with the substrate, HCO3-.

A complex of carbonic anhydrase (CA) with one of its substrates, bicarbonate, has been studied crystallographically. Human isoenzyme II was mutated at position 200 from threonine to histidine, which results in higher affinity for bicarbonate. The HCO3- ion binds in the active site to the zinc ion as a pseudo-bidentate ligand which gives the metal a coordination geometry between tetrahedral and trigonal bipyramide. The water/hydroxide normally bound with tetrahedral coordination to the zinc is probably replaced by the OH group of the bicarbonate ion. The importance of residues Thr-199 and Glu-106 in controlling the binding orientation of HCO3- is discussed as well as the catalytic mechanism. Both the complex as well as the uncomplexed mutant were studied at 1.9 A resolution.

Bicarbonates↗

Structural analysis of the zinc hydroxide-Thr-199-Glu-106 hydrogen-bond network in human carbonic anhydrase II.

The significance of the zinc hydroxide-Thr-199-Glu-106 hydrogen-bond network in the active site of human carbonic anhydrase II has been examined by X-ray crystallographic analyses of site-specific mutants. Mutants with Ala-199 and Ala-106 or Gln-106 have low catalytic activities, while a mutant with Asp-106 has almost full CO2 hydration activity. The structures of these four mutants, as well as that of the bicarbonate complex of the mutant with Ala-199, have been determined at 1.7 to 2.2 A resolution. Removal of the gamma atoms of residue 199 leads to a distorted tetrahedral geometry at the zinc ion, and a catalytically important zinc-bound water molecule has moved towards Glu-106. In the bicarbonate complex of the mutant with Ala-199 one oxygen atom from bicarbonate binds to zinc without displacing this water molecule. Tetrahedral coordination geometries are retained in the mutants at position 106. The mutants with Ala-106 and Gln-106 have a zinc-bound sulfate ion, whereas this sulfate site is only partially occupied in the mutant with Asp-106. The hydrogen-bond network seems to be "reversed" in the mutants with Ala-106 and Gln-106. The network is preserved as in native enzyme in the mutant with Asp-106 but the side chain of Asp-106 is more extended than that of Glu-106 in the native enzyme. These results illustrate the importance of Glu-106 and Thr-199 for controlling the precise coordination geometry of the zinc ion and its ligand preferences which results in an optimal orientation of a zinc-bound hydroxide ion for an attack on the CO2 substrate.

Carbonic Anhydrases↗

Acquired amegakaryocytic thrombocytopenic purpura with a Philadelphia chromosome.

A 30-year-old Chinese man with acquired amegakaryocytic thrombocytopenic purpura (AATP) and a Ph chromosome is reported. At presentation, he had severe thrombocytopenia resulting in epistaxis, gingival bleeding, and ecchymoses, while other hematologic values were within the normal range. Bone marrow aspiration showed no megakaryocytes, with a normal appearance of erythroblastic and granulopoietic series. He failed to respond to prednisone treatment, and underwent a progress from isolated thrombocytopenia to full pancytopenia. At last he died of spontaneous intracranial hemorrhage. An in vitro culture for granulocyte-macrophage precursors showed very few colonies. Karyotypic analysis revealed a standard Ph chromosome translocation, t(9;22)(q34;q11), in the majority of bone marrow cells. Southern blot analysis using a 3' bcr-HE probe didn't detect a rearrangement within the bcr DNA sequence. This patient, in fact, was a myelodysplastic disorder, initially presenting as AATP. The diagnosis of chronic myelogenous leukemia was excluded on the basis of clinical and hematologic findings. The heterogeneity of Ph chromosome in myelodysplastic syndrome is discussed.

Adult↗

Characterization of recombinant human acidic fibroblast growth factor: comparative studies with bovine acidic fibroblast growth factor.

Recombinant human acidic fibroblast growth factor (haFGF) was purified from E. coli lysate by heparin-sepharose affinity chromatography. The purified haFGF exhibited potent mitogenic activity in stimulating DNA synthesis in 3T3 cells and this activity could be significantly increased by heparin. By analysis of mitogenic activity and immunological properties, a marked difference was found between haFGF and bovine aFGF (baFGF). The main difference was that the heparin-dependence of haFGF was stronger than that of baFGF.

3T3 Cells↗

Chronic tonsillitis and IgA nephropathy.

The authors studied the effect of tonsillectomy for IgA nephropathy and the relationship between chronic tonsillitis and IgA nephropathy. Forty-five patients with IgA nephropathy were treated by tonsillectomy in our department. The effective rate of hematuria type IgA nephropathy was 86.2% and that of non-hematuria type 62.5%. We also determined the immunoglobulin composition of the tonsilla tissue in 18 IgA nephropathy patients and compared it with that of 18 chronic tonsillitis patients. The immunoglobin level of the IgA nephropathy group was higher than that of the contrast group, indicating that the disorder of tonsilla immunity may be one of the pathogenic factors for IgA nephropathy. Since no specific method has been available for the treatment of IgA nephropathy, we use tonsillectomy as an effective procedure.

Adolescent↗

Expression of a fusion protein containing calf prochymosin B(1-161) and human proinsulin.

The plasmid pJG202 containing Tac promoter, calf prochymosin B(1-161) gene and human proinsulin gene was constructed and transformed into E. coli JM105. The expression of the fusion protein consisting of 249 amino acids was controlled by IPTG and temperature, and the expressed protein was estimated to be 20-35% of the total cellular proteins by scanning of the SDS-PAGE gel stained with Coomassie brilliant blue R250. After CNBr cleavage, sulfitolysis, partial separation of the S-sulfonated human proinsulin and recombination of the disulfide bonds, human proinsulin with native properties, as evidenced by amino acid composition analysis, receptor binding and radioimmunoassays, could be obtained.

Animals↗

Specific chromosomal translocations and therapy-related leukemia induced by bimolane therapy for psoriasis.

This paper reports for the first time results of cytogenetic studies on 14 consecutive secondary acute non-lymphocytic leukemia (S-ANLL) induced by bimolane therapy. They included 10 males and 4 females with ages ranging from 17 to 54 years. They had all suffered from psoriasis and received bimolane treatment before the occurrence of their leukemia. The total dose of bimolane ranged from 40 to 400 g (mean dose 194 g). The interval between the initiation of bimolane therapy and the diagnosis of leukemia was 12-96 months (median 30 months). A preleukemic phase was only found in one case. No dysplastic features in the hemopoietic series were seen in any patient. Chromosome analysis of bone marrow cells using banding techniques revealed clonal karyotypic abnormalities in all cases: t(15;17) in 8 cases of M3, of which 75% had extra abnormalities, t(8;21) in 4 cases of M2, del(7q) only in one case of M4 and one case of M5. After antileukemic therapy, complete remission was obtained in 10 out of 12 cases with specific translocations and one out of 2 cases with 7q-anomaly, respectively. The former survived 4-58 months (median 12 months), while the latter 1 and 9 months, respectively. This study indicates that: (1) bimolane is a causative factor of leukemia in this series; (2) the leukemia in our series is therapy-related leukemia (TRL) rather than de novo ANLL; (3) there exists, in fact, a new subgroup of TRL characterized by specific rearrangements, whose clinical, hematological and prognostic features and pathogenetic mechanism may be different from classical TRL characterized by chromosome abnormalities involving absence or deletion of parts of chromosome 5 and/or 7.

Adolescent↗