Biomedical subjects

Y Ying

Publications and source records attributed to Y Ying.

At least 19 recordsLinked to original sources

PubMed2005

Sustained and stable hematopoietic donor-recipient mixed chimerism after unrelated cord blood transplantation for adult patients with severe aplastic anemia.

We evaluated the engraftment of donor cells from unrelated cord blood into adult patients with severe aplastic anemia (SAA) and the outcome of allo-CBSCT (cord blood stem cell transplantation). Nine patients were conditioned with decreased dosage of immunosuppressive agents of CTX (60 mg/kg) and ALG (120 mg/kg). The prophylaxis of GVHD consisted of standard CsA and MTX. Patients have a media age of 25.3 yr (range: 15-37), and a median weight of 57.2 kg (range: 52.5-60) at the time of transplantation. Cord blood searches were all conducted at Guangzhou Cord Blood Bank. The engraftment state of the donor cells into recipients was confirmed by microsatellite DNA fingerprinting and fluorescent quantitative PCR analysis. Engrafted evidence has been found in seven patients involved by biomolecular analyses showing donor-recipient mixed chimerism post-transplant which was stable and persistent. After a median follow up of 32.2 months (range: 4-69), seven patients were alive and disease free. This study shows that durable donor-recipient stable mixed chimerism can be achieved by unrelated CBSCT in patients with SAA. Umbilical cord blood could be employed as a source of hematopoietic stem cell for adult transplantation.

Adolescent↗

PubMed2004

Umbilical cord blood transplant for adult patients with severe aplastic anemia using anti-lymphocyte globulin and cyclophosphamide as conditioning therapy.

Allo-CBSCT (cord blood stem cell transplant) has been applied in six adult patients with severe aplastic anemia (SAA). Anti-lymphocyte globulin (ALG) 40 mg kg(-1) d(-1) x 3 days combined with cyclophosphamide (CTX) 20 mg kg(-1) d(-1) x 3 days constituted a lower intensive conditioning regimen. The prophylaxis of GVHD consisted of standard CsA and MTX. Patients are all male having a mean age of 26.5 years (range 22-38), and a median weight of 55.6 kg (range 52-60 kg). Cord blood searches were all conducted at Guangzhou Cord Blood Bank. Three of six patients in our study received one unit of cord blood in a procedure, whereas for another three patients, two units of cord blood (double units) were infused at the same time in a transplant protocol. The nine units of umbilical cord blood (UCB) infused contained 1.6-10.7 x 10(7) nucleated cells/kg body weight of the recipient after thawing. HLA antigens were identical in one unit, 1 antigen mismatched in seven, 2 antigens mismatched in 1. As of February 2003, after a median follow up of 20 months (range 7-50), four patients are alive and disease free. Five patients engrafted with molecular biology analyses showing donor-recipient mixed chimerism post transplant which is stable and persistent. One patient died of severe infection in the third month from transplant and another patient died in the early stage post transplant of serious aspergillus infection without evidence of engraftment.

Adult↗

PubMed2001

Induction of primordial germ cells from murine epiblasts by synergistic action of BMP4 and BMP8B signaling pathways.

Extraembryonic ectoderm-derived factors instruct the pluripotent epiblast cells to develop toward a restricted primordial germ cell (PGC) fate during murine gastrulation. Genes encoding Bmp4 of the Dpp class and Bmp8b of the 60A class are expressed in the extraembryonic ectoderm and targeted mutation of either results in severe defects in PGC formation. It has been shown that heterodimers of DPP and 60A classes of bone morphogenetic proteins (BMPs) are more potent than each homodimers in bone and mesoderm induction in vitro, suggesting that BMP4 and BMP8B may form heterodimers to induce PGCs. To investigate how BMP4 and BMP8B interact and signal for PGC induction, we cocultured epiblasts of embryonic day 6.0--6.25 embryos with BMP4 and BMP8B proteins produced by COS cells. Our data show that BMP4 or BMP8B homodimers alone cannot induce PGCs whereas they can in combination, providing evidence that two BMP pathways are simultaneously required for the generation of a given cell type in mammals and also providing a prototype method for PGC induction in vitro. Furthermore, the PGC defects of Bmp8b mutants can be rescued by BMP8B homodimers whereas BMP4 homodimers cannot mitigate the PGC defects of Bmp4 null mutants, suggesting that BMP4 proteins are also required for epiblast cells to gain germ-line competency before the synergistic action of BMP4 and BMP8B.

Animals↗

PubMed2001

Thermolysis of free-radical initiators: tert-butylazocumene and its 1,3- and 1,4-bisazo and 1,3,5-trisazo analogues.

Four tert-butylazocumenes (4-7) were prepared from the corresponding cyanobenzenes, and their nitrogen evolution kinetics and products were analyzed. In combination with TEMPO, the simplest compound, tert-butylazocumene (4), shows promise as a "one-radical" initiator of styrene polymerization. The ABNO-trapped cumyl radical 29 is a particularly stable trialkylhydroxylamine, whose thermolysis half-life is 2.1 h at 150 degrees C. Taking advantage of this stability, we trapped the cumyl radical centers from 7 to afford tris adduct 32a. While the behavior of the meta bisazoalkane 6 can be mostly predicted from that of 4, the para isomer 5 exhibits both unusual products and kinetics, attributed to the formation of quinodimethane 33 via azo-containing radical 34. In fact, flash vacuum pyrolysis of 5 allowed observation of the (1)H and (13)C NMR spectra of 33, whose persistence even at ambient temperature showed that this quinodimethane is far more stable than the parent 36. Finally, evidence is presented that 7 is an initiator of star polymerization.

Journal Article↗

PubMed2001

Cooperation of endoderm-derived BMP2 and extraembryonic ectoderm-derived BMP4 in primordial germ cell generation in the mouse.

The primordial germ cells (PGCs) of the mouse are derived from proximal epiblast cells that are adjacent to the extraembryonic ectoderm during gastrulation. Previous studies have demonstrated that extraembryonic ectoderm-derived BMP4 and BMP8B are both required for PGC generation. Here we show that Bmp2, a member of the Dpp class of the Bmp superfamily, also plays a role in PGC generation. PGC number is significantly reduced in Bmp2 heterozygous and homozygous embryos at the N2 generation onto C57BL/6 background. Bmp2 homozygous embryos also have a short allantois and about 50% of them do not undergo normal chorioallantoic fusion. Using whole-mount in situ hybridization, we show that Bmp2 is primarily expressed in the endoderm of mouse pregastrula and gastrula embryos. Using a genetic approach, we further show that Bmp2 and Bmp4, but not Bmp2 and Bmp8b, have an additive effect on PGC generation. These results suggest that PGC generation in the mouse embryo is regulated not only by extraembryonic ectoderm-derived BMP4 and BMP8B, but also by endoderm-derived BMP2.

Allantois↗

PubMed2000

Synaptotagmin regulation of coated pit assembly.

Synaptotagmins bind clathrin AP-2 with high affinity via their second C(2) domain, which indicates they are involved in coated pit function. We now report that expression of synaptotagmins lacking either the second C(2) domain or the entire cytoplasmic region potently inhibit endocytosis. Inhibition was dependent on two intramembrane cysteine residues that were found to be essential for synaptotagmin oligomerization. Cells expressing the wild-type, but not the mutant, truncated synaptotagmin fragment had a reduced number of clathrin-coated pits. These results suggest that the formation of synaptotagmin multimers is an important step in the regulation of coated pit assembly.

Adaptor Protein Complex alpha Subunits↗

PubMed2000

Synergistic effects of DL111-IT combined with mifepristone on termination of early pregnancy in rhesus monkeys.

The objectives of this study were to determine the synergistic effects of DL111-IT in combination with mifepristone (RU486) on termination of early pregnancy in rhesus monkeys. Pregnancy was confirmed by tactile sensation of pregnant uterus via anus with finger and ultrasound examination. Pregnancy termination was obtained with vaginal bleeding and abortion materials including fetuses and placentae after treatment. With multiple doses of DL111-IT or RU486 given alone between d24 and d50 of gestation, pregnancy arrests were obtained in 40% (2/5) of monkeys treated with DL111-IT intramuscularly (im) (25 mg x kg(-1) x d(-1) x 3 days), in 20% (1/5) of monkeys treated with 9 mg x kg(-1) x d(-1) x 2 days, and 4.5 mg x kg(-1) on day 3 with RU486 intragastrically (ig). DL111-IT (25 mg x kg(-1) on day 1, im) in combination with RU486 (the same treatment as above) resulted in 100% (10/10) termination of pregnancy and uterine bleeding lasted 6.6 +/- 1.3 days. RU486 (as above treatment) in combination with misoprostal (Miso, 109 microg x kg(-1) on day 3, ig) showed 71.4% (5/7) termination of pregnancy, and uterine bleeding lasted 12.9 +/- 9.6 days. The synergistic effect of DL111-IT plus RU486 enhances termination of early pregnancy and significantly shortens the bleeding time than RU486 plus Miso does in rhesus monkeys.

Abortifacient Agents, Nonsteroidal↗

PubMed2000

Molecular cloning and expression analysis of a mouse UDP-GlcNAc:Gal(beta1-4)Glc(NAc)-R beta1,3-N-acetylglucosaminyltransferase homologous to Drosophila melanogaster Brainiac and the beta1,3-galactosyltransferase family.

We have isolated a murine cDNA coding for a beta1,3-N-acetylglucosaminyltransferase enzyme ( beta3GnT). This enzyme is similar in sequence to Drosophila melanogaster Brainiac and to the murine and human beta1,3-galactosyltransferase family of proteins. The mouse beta 3GnT protein is 397 amino acids in length and contains 7 cysteine residues that are conserved in the human orthologue. beta 3GnT is a type II membrane protein localized to the Golgi apparatus. Enzyme assays with recombinant mouse beta 3GnT reveal that it has a preference for acceptors with Gal(beta1-4)Glc(NAc) at the non-reducing termini. Proton NMR analysis of product showed incorporation of GlcNAc in beta1,3 linkage to the terminal Gal of Gal(beta1-4)Glc(beta1-O-benzyl). Northern blot analysis revealed the presence of a single 3.0[emsp4 ]kb transcript in all adult mouse and human organs tested, with highest levels in the kidney, liver, heart and placenta. The beta 3GnT gene is also expressed in a number of tumor cell lines. The human orthologue of beta 3GnT is located on chromosome 2pl5.

Amino Acid Sequence↗

PubMed2000

Umbilical cord blood transplantation from unrelated HLA-matched donor in an adult with severe aplastic anemia.

A 23-year-old male suffering from severe aplastic anemia (SAA) weighing 60 kg was successfully treated by unrelated allo-CBSCT (cord blood stem cell transplantation). A six-loci HLA-identical umbilical cord blood (UCB) was infused after conditioning with low-dose cyclophosphamide (CTX) and antilymphocyte globulin (ALG). The prophylaxis of GVHD consisted of CsA and MTX. The infused cord blood provided 1.89 x 10(7) nucleated cells per kg, CD34-positive cells: 0.89%. Neutrophils >0.5 x 10(9)/l were reached 10 days after transplant, and platelets greater than 50.0 x 10(9)/l at day 26. RBC and platelet transfusion independence were reached on days 15 and 18. The patient developed grade 1 skin GVHD 10 months after engraftment of the donor cells. Microsatellite DNA fingerprinting indicated a stable and persistent donor-recipient mixed chimerism, whilst the circulating red cells remain of host origin.

Adult↗

PubMed2000

Detection of multiple bone morphogenetic protein messenger ribonucleic acids and their signal transducer, Smad1, during mouse decidualization.

Decidualization is a process characterized by morphological and functional changes in the uterine stromal cells. In addition to steroid hormones, growth factors are implicated in this process. Using in situ hybridization, we found that mRNAs for several bone morphogenetic proteins (BMPs) were detected in the decidual and vascular endothelial cells. The Bmp7 mRNA was detected in the decidualizing stromal cells surrounding the blastocyst and distributed in a gradient, with the highest levels occurring near the uterine epithelium at 4.5 days post-coitus (dpc). With the progression of decidualization, Bmp7 signals in the deciduum at the antimesometrial side decreased, but strong signals were retained in the decidual area at the mesometrial side at 7.0 dpc. In contrast, Bmp8a transcripts increased from 5.5 to 7.0 dpc in the decidual tissue, with the highest levels occurring in the secondary decidual zone at the antimesometrial side. The Bmp2, Bmp4, and Smad1 transcripts were found in the secondary decidual zone, especially at the mesometrial side. The Bmp2 signals were primarily detected in decidual cells, whereas Bmp4 and Smad1 transcripts were mainly detected in vascular endothelial cells, suggesting that they may be involved in decidual angiogenesis.

Animals↗

PubMed2000

Requirement of Bmp8b for the generation of primordial germ cells in the mouse.

In the mouse embryo, the generation of primordial germ cells (PGCs) from the epiblast requires a bone morphogenetic protein-4 (BMP4) signal from the adjacent extraembryonic ectoderm. In this study, we report that Bmp8b, a member of the Gbb-60A class of the BMP superfamily, is expressed in the extraembryonic ectoderm in pregastrula and gastrula stage mouse embryos and is required for PGC generation. A mutation in Bmp8b on a mixed genetic background results in the absence of PGCs in 43% null mutant embryos and severe reduction in PGC number in the remainder. The heterozygotes are unaffected. On a largely C57BL/6 background, Bmp8b null mutants completely lack PGCs, and Bmp8b heterozygotes have a reduced number of PGCs. In addition, Bmp8b homozygous null embryos on both genetic backgrounds have a short allantois, and this organ is missing in some more severe mutants. Since Bmp4 heterozygote embryos have reduced numbers of PGCs, we used a genetic approach to generate double-mutant embryos to study interactions of Bmp8b and Bmp4. Embryos that are double heterozygotes for the Bmp8b and Bmp4 mutations have similar defects in PGC number as Bmp4 heterozygotes, indicating that the effects of the two BMPs are not additive. These findings suggest that BMP4 and BMP8B function as heterodimers and homodimers in PGC specification in the mouse.

Animals↗

PubMed2000

[Umbilical cord blood stem cells transplantation from an unrelated donor into an adult with severe aplastic anemia].

OBJECTIVE: To evaluate the availability of the treatment of adult severe aplastic anemia with unrelated allo-cord blood stem cell transplantation (CBSCT). METHODS: HLA-matched unrelated cord blood transplantation has been successfully performed for an adult severe aplastic anemia patient. A unit of cord blood provided by Guangzhou Cord Blood Bank containing 1.89 x 10(7)/kg mononucleated cells, 1.8 x 10(4)/kg CFU-GM and of CD(34) positive cells was 0.009. The patient was conditioned with CTX (60 mg/kg) and anti-lymphocyte globulin (ALG, 120 mg/kg). GVHD prophylaxis consisted of both MTX and CsA. The CsA had been given for 8 months. RESULTS: The lowest ANC was 0.6 x 10(9)/L post-transplantation. The patient achieved an ANC of greater than 0.5 x 10(9)/L at 10 days, and the platelet of greater than 50.0 x 10(9)/L at day 20 after transplantation. He developed Grade 1 GVHD in the tenth month after grafting. Microsatellite DNA fingerprinting indicated a stable donor-recipient mixed chimerism, whilst the circulating red cells remained host origin. CONCLUSION: It is the first report in China on successful treatment of adult severe aplastic anemia by unrelated allo-CBSCT.

Adult↗

PubMed1999

Effects of male accessory sex glands on sperm decondensation and oocyte activation during in vivo fertilization in golden hamsters.

Removal of paternal male accessory sex glands (ASG) could cause a delay in DNA synthesis in hamster zygotes fertilized in vivo. In view of the fact that this process is closely related to pronuclear development which, in part, depends on sperm nuclear decondensation and oocyte activation during fertilization, we carried out a series of experiments were undertaken to determine whether ASG also has an effect on these early events. (1) Oocytes were collected from females mated with SH (sham-operated control), AGX (bilateral excision of ampullary glands), VPX (bilateral excision of ventral prostates) or TX (excision of all ASG) males (n = 8 per group) at 4, 5 and 6 h post coitus. (2) Epididymal spermatozoa were incubated with total ventral prostate (VP) secretion to study its effect on dithiothreitol-induced sperm decondensation. (3) Histone H1 kinase activity in oocytes collected as described in (1) was determined. (4) Exocytosed cortical granules on oocytes were labelled with FITC-LCA and quantified by a Metamorph Imaging System. Results showed that sperm decondensation and resumption of meiosis in oocytes in VPX and TX groups were significantly slower compared with SH. VP secretion augmented sperm decondensation in vitro. At 4 h post coitus, the relative activity of histone H1 kinase in the TX and VPX groups was significantly higher than that in the SH group (p < 0.01). Cortical granule exocytosis in the AGX group was consistently weaker at all time points studied and was significantly lower than that of the control at 4 h post coitus (p < 0.05), while the percentage of polyspermic fertilization in the AGX group was significantly higher compared with that in the SH group (p < 0.05). Taken together, these results show that the lack of exposure of spermatozoa to secretions of the ASG does not jeopardize their ability to penetrate ova, although other aspects of their function in the early stages of gamete interaction and subsequent initiation of embryonic development are affected.

Animals↗

PubMed1999

Male accessory sex gland secretions affect oocyte Ca2+ oscillations during in-vitro fertilization in golden hamsters.

To evaluate the effect of male accessory sex gland secretions on Ca2+ oscillations of oocytes, epididymal or ejaculated spermatozoa recovered from uteri were used to inseminate oocytes. Ca2+ oscillations were measured by Fura 2 fluorescence imaging (F340/F380). We showed that although Ca2+ oscillations induced by ejaculated spermatozoa had a pattern similar to those induced by epididymal spermatozoa, the amplitude of the first Ca2+ transient in the former group was significantly higher (P < 0.05) and the duration was significantly longer (P < 0.01). Oocytes inseminated with ejaculated spermatozoa recovered from uteri from males had ampullary glands or ventral prostates removed showed significantly lower Ca2+ oscillations compared to the controls (P < 0.05, P < 0.01 respectively). Moreover, the relative area of the first Ca2+ transient in treatment groups was significantly smaller than the control. In addition, a significantly higher percentage of oocytes (52%) inseminated by spermatozoa from males with all accessory sex glands removed showed non-oscillatory Ca2+ transients, compared to the controls (5%, P < 0.05). These results indicate that accessory sex gland secretions can affect Ca2+ oscillations. The differences between Ca2+ oscillations induced by epididymal and uterine spermatozoa from males with all accessory sex glands removed suggest that uterine factors may also influence this process.

Animals↗

PubMed1999

Placebo-controlled trial of 13-cis-retinoic acid activity on retinoic acid receptor-beta expression in a population at high risk: implications for chemoprevention of lung cancer.

PURPOSE: To establish the incidence of abnormalities in the expression of retinoic acid receptor-beta (RARbeta) in bronchial cells and determine the capacity of 13-cis-retinoic acid (13-CRA) to correct such abnormalities. PATIENTS AND METHODS: One hundred eighty-eight smokers had a medical indication for bronchoscopy and were studied with bronchial brushings. Bronchial brushing samples were obtained for cytology analysis and for molecular analysis. After RNA was extracted, RARbeta sequences were amplified by reverse transcriptase polymerase chain reaction and Southern blots were performed to assess RARbeta expression. Forty-four eligible individuals with diminished RARbeta expression consented to double-blind randomization to receive a placebo or 13-CRA 30 mg orally daily for 6 months. A second bronchoscopy was performed at the end of the treatment period. An analysis of variance was used to analyze changes in RARbeta expression before and after treatment. RESULTS: The 6-month treatment course was completed by 27 patients, and results were obtained for a total of 18 patients (eight patients treated with 13-CRA and ten treated with the placebo). In the placebo group, there was no difference between the results of RARbeta expression before and after treatment (P =.43). In the 13-CRA group, there was an upregulation of RARbeta expression at the end of 13-CRA treatment (P =.001). Cytologic changes were uncommon. Toxicities were primarily of grade 1. Palatal brushings were compared with bronchial brushings in 40 smokers. A perfect correlation of the results of RARbeta expression was obtained from 27 patients. CONCLUSION: RARbeta expression is frequently decreased in the bronchial epithelium of smokers and is upregulated at the end of 13-CRA treatment. These results support undertaking a phase III chemoprevention trial of 13-CRA treatment for lung cancer.

Analysis of Variance↗

PubMed1999

[DNase I sensitivity of nucleus and chromatin in six different tissues of old rates].

In order to investigate the mechanism of aging, we studied the DNase I sensitivity of nucleus and chromatin in six different tissue (heart, liver, spleen, lung, kidney and brain of old and weanling wistar rats. The results showed that the DNase I digestive sensitivity levels of nucleus and chromatin in the tissue of the old rats were lower than those of the weanling rats respectively that in the old rats, the DNase I digestive sensitively in the tissue varied, the lowest sensitivity was in brain (P < 0.01); and that in the weanling rats, lowest DNase I digestive sensitivity of nucleus and chromatin was also in the brain, but the difference was not significant.

Aging↗

PubMed1998

Annexin VI-mediated loss of spectrin during coated pit budding is coupled to delivery of LDL to lysosomes.

Previously we reported that annexin VI is required for the budding of clathrin-coated pits from human fibroblast plasma membranes in vitro. Here we show that annexin VI bound to the NH2-terminal 28-kD portion of membrane spectrin is as effective as cytosolic annexin VI in supporting coated pit budding. Annexin VI-dependent budding is accompanied by the loss of approximately 50% of the spectrin from the membrane and is blocked by the cysteine protease inhibitor N-acetyl-leucyl-leucyl-norleucinal (ALLN). Incubation of fibroblasts in the presence of ALLN initially blocks the uptake of low density lipoprotein (LDL), but the cells recover after 1 h and internalize LDL with normal kinetics. The LDL internalized under these conditions, however, fails to migrate to the center of the cell and is not degraded. ALLN-treated cells have twice as many coated pits and twofold more membrane clathrin, suggesting that new coated pits have assembled. Annexin VI is not required for the budding of these new coated pits and ALLN does not inhibit. Finally, microinjection of a truncated annexin VI that inhibits budding in vitro has the same effect on LDL internalization as ALLN. These findings suggest that fibroblasts are able to make at least two types of coated pits, one of which requires the annexin VI-dependent activation of a cysteine protease to disconnect the clathrin lattice from the spectrin membrane cytoskeleton during the final stages of budding.

Annexin A6↗

PubMed1998

Characterization of a cytosolic heat-shock protein-caveolin chaperone complex. Involvement in cholesterol trafficking.

Caveolin is a 22-kDa protein that appears to play a critical role in regulating the cholesterol concentration of caveolae. Even though caveolin is thought to be a membrane protein, several reports suggest that this peculiar protein can traffic independently of membrane vesicles. We now present evidence that a cytosolic pool of caveolin is part of a heat-shock protein-immunophilin chaperone complex consisting of caveolin, heat-shock protein 56, cyclophilin 40, cyclophilin A, and cholesterol. Treatment of NIH 3T3 cells with 1 microM cyclosporin A or 100 nM rapamycin disrupted the putative transport complex and prevented rapid (10-20 min) transport of cholesterol to caveolae. The lymphoid cell line, L1210-JF, does not express caveolin, does not form an immunophilin-caveolin complex, and does not transport newly synthesized cholesterol to caveolae. Transfection of caveolin cDNA into L1210-JF cells allowed the assembly of a transport complex identical to that found in NIH 3T3 cells. In addition, newly synthesized cholesterol in transfected cells was rapidly (10-20 min) and specifically transported to caveolae. These data strongly suggest that a caveolin-chaperone complex is a mechanism by which newly synthesized cholesterol is transported from the endoplasmic reticulum through the cytoplasm to caveolae.

Animals↗