OBJECTIVETo investigate the relationship between human embryo implantation rates and the zygote morphology and establish zygote morphologic indices that indicate the embryo implantation potential after in vitro fertilization and embryo transfer (IVF-ET).

METHODSSixty-two patients with IVF-ET were enrolled in this study, who were below 35 years of age with endometrium thickness no greater than 8 mm on the day of HCG injection. Embryo transfer was performed on day 3 after oocyte retrieval, and 30 patients with successful implantation of all the embryos transferred (implantation rate of 100%) were allocated into the implantation group, and the other 32 patients with implantation failure (implantation rate of 0) served as the control group. The zygote morphologic characteristics were analyzed for the pronuclei, nucleolar precursor bodies (NPB), polar body, cytoplasmic halo, color and granulation of the cytoplasm, and the results were compared between the two groups.

RESULTSThe implantation rate was significantly higher for embryos with the two pronuclei in close vicinity, central position of the pronuclei in the cytoplasm and comparable size of the two pronuclei. Embryos developed from zygotes with linear arrangement of 3 to 7 NPB in moderate sizes tented to have a higher implantation rate (P<0.05). The implantation rate could be obviously lowered when the cytoplasm contained excessive cytoplasmic granularity (P<0.05). The other morphologic characteristics of the embryos such as the polar bodies, color of the cytoplasm, cytoplasm halo, or vacuoles in the cytoplasm did not significantly impact on the implantation rate.

CONCLUSIONThe morphology of the two pronuclei reflects the quality of the zygote and may help predict the developmental potential of the embryo chosen for transfer on day 3 in IVF.

Adult ; Cell Nucleolus ; metabolism ; Cytoplasmic Granules ; metabolism ; Embryo Implantation ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Zygote ; cytology

Nucleophosmin (NPM) is a protein that shuttles between the nucleus, nucleoplasm and cytoplasm. NPM gene mutations and aberrant cytoplasmic NPM localization have been recently described in acute myelogenous leukemia (AML) with normal karyotype and in a few myelodysplastic syndromes. Expression of NPM mutant reduces the ability of Arf to initiate a p53 response and to induce cell cycle arrest. Clinical research has revealed that NPM mutations are relative to prognosis and can be used to monitor and quantify minimal residual disease (MRD) in AML patients with normal karyotype, therefore, these findings indicate that nucleophosmin mutations might contribute to illustration of myeloid leukemogenesis. In this paper, the research progress of nucleophosmin mutations in haematological malignancies was reviewed.
Cell Nucleolus ; metabolism ; Hematologic Neoplasms ; genetics ; pathology ; Humans ; Mutation ; Nuclear Proteins ; genetics ; fms-Like Tyrosine Kinase 3 ; genetics

Aneuploidy ; Biomarkers, Tumor ; metabolism ; DNA, Neoplasm ; genetics ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Nucleolus Organizer Region ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; S Phase

OBJECTIVE: To investigate the effect of oxidative stress on the accumulation of heat shock protein 70 (HSP70) within C2C12 myogenic cells. METHODS: Heat shock response (42 degrees C for 1 h and recovery for 12 h at 37 degrees C) was used to induce the expression of heat shock protein 70. We constructed a recombinant plasmid of HSP70 with enhanced green fluorescent protein (EGFP). After being transfected transiently into C2C12 cells, immunoblotting was used to detect the expression of HSP70 induced by heat shock response and transfection. Immunocytochemistry, fluorescent microscopy and immunoblotting were used to detect the translocation of HSP70. RESULTS: Immunoblotting showed that the overexpression of HSP70 was induced by heat shock response and transient transfenction. HSP70 localized within the cytoplasm of the normal cells, but HSP70 translocated from the cytoplasm to the nucleus and the nucleolus at 1 h after the treatment of oxidative stress (0.5 mmol/L H2O2) by using immunocytochemistry, fluorescent microscopy and immunoblotting for cellular partial proteins. CONCLUSION Oxidative stress may induce the accumulation of heat shock protein 70 within the nucleolus.
Cell Nucleolus ; metabolism ; Cells, Cultured ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Myoblasts ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Oxidative Stress ; physiology

Since there have been very few studies on nucleolar signaling, an attempt was made to establish nucleolar signal pathways which link the cell membrane to the nucleolus for the transfer of extracellular signals. Two pathways were studied. One was the G alpha s mediated cAMP pathway where two signal molecules were yielded, including RII and protein kinase A. The other was the G alpha q mediated DAG/IP3 pathway which yields two signals including protein kinase C and IP3/Ca2+. By the studying isolated nucleoli from resting liver, regenerating liver or weak carcinogen thioacetamide treated liver, it was possible to detect protein kinase A (PKA), protein kinase C (PKC) and RII subunits. In addition, CK2 was detected. It was found that external signals transmitted through G protein coupled receptors could reach into the nucleolus and that physical translocation of signal molecules was an integral step involved in membrane-nucleolus linked pathways. When an in vitro assay of the above signal molecules was carried out using [gamma-32P]-ATP, most kinase dependent phosphorylation was via the major CK2 (more than 95%). Therefore, it is suggested that the major CK2 dependent pathway is involved in 'house keeping' for nucleolar integrity and the minor pathways, dependent on PKA, PKC and others, are involved in subtle regulatory mechanisms such as 'extra-house-keeping' activities by nucleolar chromosomal remodeling.
Animal ; Blotting, Western ; Cell Membrane/metabolism ; Cell Nucleolus/metabolism* ; Cyclic AMP-Dependent Protein Kinases/metabolism ; GTP-Binding Proteins/metabolism ; Immunoblotting ; Liver/metabolism ; Liver Neoplasms, Experimental ; Male ; Models, Biological ; Nuclear Proteins/metabolism* ; Phosphoproteins/metabolism ; Protein Kinase C/metabolism ; Protein-Serine-Threonine Kinases/metabolism ; Rats ; Rats, Sprague-Dawley ; Second Messenger Systems ; Signal Transduction* ; Thioacetamide/pharmacology ; Time Factors

OBJECTIVETo identify the putative specific localization signal sequence of tumor metastasis suppressor gene-1 (TMSG-1) and to explore the mechanism of subcellular localization of TMSG-1 protein.

METHODSVectors expressing green fluorescence protein (GFP) tagged different TMSG-1 fragments were generated and transfected into human embryo kidney 293 (HEK293) cells. The expression of those fusion proteins was detected by Western blotting and their subcellular localizations were observed by laser confocal microscope.

RESULTSGFP was fused with the native TMSG-1(aa1-380) or different fragments including T1 (aa1-70), T2 (aa1-128), T3 (aa129-380), T4 (aa71-128), T5 (aa71-179) and T6 (aa71-380). Anti-GFP Western blotting showed that these fusion proteins were successfully expressed. Under laser confocal microscope, GFP fused with fragment T4 (aa71-128) localized mainly in the nucleolus; GFP fused with fragment T6 (aa71-380) localized diffusely in the nucleus; while other fusion proteins with TMSG-1 (aa1-380) or fragment T1 (aa1-70), T2 (aa1-128), T3 (aa129-380) and T5 (aa71-179) localized in the cytoplasm. Fragment T4(Δ119-128) was generated from T4 with deletion of 10 amino acid of the C terminal. GFP fused with fragment T4(Δ119-128) remained in the nucleus, but no longer in the nucleolus.

CONCLUSIONSThere is a nucleolar localization signal (aa119-128 RRRRNQDRPS) within TMSG-1. This finding may have laid the foundation for further investigations into subcellular localization and function of TMSG-1.

Amino Acid Sequence ; Blotting, Western ; Cell Nucleolus ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Membrane Proteins ; genetics ; metabolism ; Microscopy, Confocal ; Nuclear Localization Signals ; Plasmids ; Recombinant Fusion Proteins ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism