PubMed Health⌕ Search

Biomedical subjects

X Shan

Publications and source records attributed to X Shan.

At least 19 recordsLinked to original sources
PubMed2006

Presynaptic localization of a truncated isoform of the type 2 corticotropin releasing factor receptor in the cerebellum.

It is now well established that corticotropin releasing factor is present in two major excitatory afferent systems to the cerebellum, namely climbing fibers and mossy fibers. Two major classes of corticotropin releasing factor receptors, each with unique binding characteristics, have been identified as type 1 and type 2. In this study we used an antibody made to the n-terminus of the type 2 corticotropin releasing factor receptor. Characterization of this antibody showed that it strongly labeled a protein with a molecular weight of 16-32 kDa and only faintly labels a 62-83 kDa protein. The lower molecular weight protein corresponds to the weight of a recently described truncated isoform of this receptor that is designated corticotropin releasing factor-type 2alpha-truncated isoform. We carried out transfection paradigms using corticotropin releasing factor-type 2alpha-truncated isoform constructs and confirmed that the antibody recognized the truncated isoform of the type 2 corticotropin releasing factor receptor. Further, light and electron microscopic studies were carried out in mice and rats to define the distribution of the truncated receptor. Immunoreactivity is evident in the basal region of many, but not all Purkinje cell bodies and their initial axonal segments, as well as the initial axonal segments of isolated Golgi cells, and cerebellar nuclear neurons. In addition, punctate elements in the molecular layer were immunolabeled. The localization of the receptor to the initial segment of Purkinje cells was confirmed with electron microscopy. Further, the punctate labeling in the molecular layer was localized to parallel fibers and their terminals. In conclusion, evidence has been presented to show that distinct isoforms of the corticotropin releasing factor receptor are present in the cerebellum. The complex interactions between corticotropin releasing factor and other members of the corticotropin releasing factor family of peptides with both pre- and postsynaptic receptors support a growing concept that corticotropin releasing factor plays an important role in modulating activity in cerebellar circuits and ultimately in controlling motor behavior.

Adrenocorticotropic Hormone↗
PubMed2005

Effects of human soluble BAFF synthesized in Escherichia coli on CD4+ and CD8+ T lymphocytes as well as NK cells in mice.

B cell-activating factor belonging to the TNF family (BAFF, also called BLyS, TALL-1, zTNF-4, or THANK) is an important survival factor for B lymphocytes. In this study, we injected mouse abdominal cavity with human soluble BAFF (hsBAFF, 0.01, 0.1, 0.5, 2 mg/kg body mass) synthesized in Escherichia coli. On the 8th day after injection, we investigated the effects of hsBAFF on immune functional activities of splenic B lymphocytes, CD4(+) and CD8(+) T lymphocytes and natural killer (NK) cells in mice. The results showed that B lymphocyte proliferation significantly increased in hsBAFF-treated groups with dosages of 0.1 mg/kg (p<0.05), 0.5 and 2 mg/kg (p<0.01). We observed a dose-dependent increase of CD4(+) T lymphocyte percentage and significantly higher values in 0.5 and 2 mg/kg hsBAFF-treated groups (p<0.05 and p<0.001, respectively) compared to control group, but CD8(+) T lymphocyte percentage remained unchanged. The ratio of CD4(+) to CD8(+) T lymphocytes rose with increasing hsBAFF dosage (p<0.05 for 2 mg/kg hsBAFF vs. control). Significantly stronger NK cell activities were found in 0.5 and 2 mg/kg hsBAFF-treated groups (p<0.05). The main finding of this study is that the hsBAFF can enhance immune responses in the body by increasing B lymphocyte and CD4(+) T lymphocyte function as well as elevating NK cell activity.

Animals↗
PubMed2005

Increased phospho-adducin immunoreactivity in a murine model of amyotrophic lateral sclerosis.

Adducins alpha, beta and gamma are proteins that link spectrin and actin in the regulation of cytoskeletal architecture and are substrates for protein kinase C and other signaling molecules. Previous studies have shown that expressions of phosphorylated adducin (phospho-adducin) and protein kinase C are increased in spinal cord tissue from patients who died with amyotrophic lateral sclerosis, a neurodegenerative disorder of motoneurons and other cells. However, the distribution of phospho-adducin immunoreactivity has not been described in the mammalian spinal cord. We have evaluated the distribution of immunoreactivity to serine/threonine-dependent phospho-adducin at a region corresponding to the myristoylated alanine-rich C kinase substrate-related domain of adducin in spinal cords of mice over-expressing mutant human superoxide dismutase, an animal model of amyotrophic lateral sclerosis, and in control littermates. We find phospho-adducin immunoreactivity in control spinal cord in ependymal cells surrounding the central canal, neurons and astrocytes. Phospho-adducin immunoreactivity is localized to the cell bodies, dendrites and axons of some motoneurons, as well as to astrocytes in the gray and white matter. Spinal cords of mutant human superoxide dismutase mice having motoneuron loss exhibit significantly increased phospho-adducin immunoreactivity in ventral and dorsal horn spinal cord regions, but not in ependyma surrounding the central canal, compared with control animals. Increased phospho-adducin immunoreactivity localizes predominantly to astrocytes and likely increases as a consequence of the astrogliosis that occurs in the mutant human superoxide dismutase mouse with disease progression. These findings demonstrate increased immunoreactivity against phosphorylated adducin at the myristoylated alanine-rich C kinase substrate domain in a murine model of amyotrophic lateral sclerosis. As adducin is a substrate for protein kinase C at the myristoylated alanine-rich C kinase substrate domain, the increased phospho-adducin immunoreactivity is likely a consequence of protein kinase C activation in neurons and astrocytes of the spinal cord and evidence for aberrant phosphorylation events in mutant human superoxide dismutase mice that may affect neuron survival.

Amyotrophic Lateral Sclerosis↗
PubMed2001

Isomer formation and other issues in the substitution reactions of oxorhenium(V) complexes of 2,2'-bipyridine and related ligands.

Two new oxorhenium(V) compounds were prepared and characterized: MeReO(mtp)(Me(2)Bpy) and MeReO(mtp)(dppb), where mtpH(2) is 2-(mercaptomethyl)thiophenol, Me(2)Bpy is 4,4'-dimethyl-2,2'-bipyridine, and dppb is 1,2-(Ph(2)P)(2)C(6)H(4). The more stable geometric isomer of MeReO(mtp)X forms MeReO(mtp)Y (X, Y = PR(3), NC(5)H(4)R) in two steps, both of which show a first-order dependence on [Y], proceeding through the metastable geometric isomer MeReO(mtp)Y. When Y = PR(3), no MeReO(mtp)Y was detected at equilibrium; with NC(5)H(4)R, however, both isomers were detected. The values of K(PyPy) were 8.5-9.8, largely irrespective of R; for NC(5)H(5), DeltaH degrees = -4.47 +/- 0.29 kJ and DeltaS degrees = 3.9 +/- 1.0 J K(-1). For the more symmetric edt ligand, geometric isomers do not exist, but enantiomers do. The rate of racemization of MeReO(edt)(NC(5)H(4)R) was proportional to [Py]. Values of k(rac) for 16 compounds span the range 135-370 L mol(-1) s(-1) in C(6)H(6) at 25 degrees C (rho = -0.39 +/- 0.07). In toluene-d(8), k(rac) for 4-picoline has DeltaH = 28.9 +/- 0.4 kJ, DeltaS() = -103.6 +/- 0.9 J K(-1). A common mechanism applies to ligand substitution (mtp) and racemization (edt). MeReO(dithiolate)Py complexes react with Bpy, Me(2)Bpy, Phen, and Me(2)Phen to form six-coordinate chelates, with rate constants 0.024-0.74 L mol(-1) s(-1) at 25 degrees C, some 10(3) times smaller than with pyridines, no doubt owing to the bulk of the bidentates. Values of DeltaS are -86 to -138 J K(-1), reflecting substantial orientational barriers as well as the inherent contribution of the associative mechanism. The product is MeReO(mtp)(Me(2)Bpy). The formation of the metastable isomer is consistent with the mechanism assigned to the ligand substitution and racemization reactions. Such compounds, once formed, no longer participate in ligand substitution reactions at reasonable rates. The formation of the metastable isomer is consistent with the mechanism assigned to the ligand substitution and racemization reactions.

Journal Article↗
PubMed2001

Luminol chemiluminescence in unbuffered solutions with a cobalt(II)-ethanolamine complex immobilized on resin as catalyst and its application to analysis.

Using a heterogeneous catalyst, Co(II)-ethanolamine complex sorbed on Dowex-50W resin, the chemiluminescence (CL) of luminol in unbuffered or weakly acidic solution was studied in the presence of H2O2. The maximum luminol CL wavelength at pH 5.7 was 448 nm, 23 nm longer than that in a basic solution (pH 10.5). Three different ligands, mono-, di-, and triethanolamine, and six transition metal ions, Co(II), Cu(II), Ni(II), Mn-(II), Fe(II), and Fe(III) were compared by CL measurements. The CL intensity decreased in the order mono- > di- > triethanolamine and Co(II) > Cu(II) > Ni(II) > Fe-(III) > Mn(II) > Fe(II). This heterogeneous CL system was developed as H2O2 and glucose flow-through sensors. Detection limits (S/N = 3) of H2O2 and glucose using Dowex-50W-X4-Co(II)-monoethanolamine as catalyst are 1 x 10(-7) M and 1 x 10(-6) M, respectively. On the basis of the studies of the CL, fluorescence, UV-vis and ESCA spectra and the effect of dissolved oxygen in luminol solution, a mechanism for CL emission in unbuffered solution was considered as the formation of a superoxide radical ion during the decomposition of H2O2 catalyzed by the Co(II)-ethanolamine immobilized resin. Then the superoxide radical ion acted on luminol and the CL was emitted. The applications of the proposed method to determine H2O2 in rainwater without any special pretreatment and glucose in human urine and orange juice samples give satisfactory results.

Anion Exchange Resins↗
PubMed2001

Mutations in the regulatory domain of cystathionine beta synthase can functionally suppress patient-derived mutations in cis.

Human cystathionine beta--synthase (CBS) is an S-adenosylmethionine-regulated enzyme that plays a key role in the metabolism of homocysteine. Mutations in CBS are known to cause homocystinuria, an inborn error in metabolism. We previously developed a yeast functional assay for CBS and used it to characterize mutations found in homocystinuric patients. We discovered that many patient-derived mutations are functionally suppressed by deletion of the C-terminal 142 amino acids, which contain a 53 amino acid motif known as the CBS domain. This domain is found in a wide variety of proteins of diverse biological function. Here we have used a genetic screen to identify missense mutations in the C-terminal region of CBS that can suppress the most common patient mutation, I278T. Seven suppressor mutations were identified, four of which map to the CBS domain. When combined in cis with another pathogenic mutation, V168M, six of seven of the suppressor mutations rescued the yeast phenotype. Enzyme activity analyses indicate that the suppressors restore activity from <2% to 17--64% of the wild-type levels. Analysis of the suppressor mutations in the absence of the pathogenic mutation shows that six of the seven suppressor alleles have lost enzymatic responsiveness to S-adenosylmethionine. Using homology modeling, we show that the suppressor mutations appear to map on one face of the CBS domain. Our results indicate that subtle changes to the C-terminus of CBS can restore activity to mutant proteins and provide a rationale for screening for compounds that can activate mutant CBS alleles.

Amino Acid Sequence↗
PubMed2001

Uptake and distribution of rare earth elements in rice seeds cultured in fertilizer solution of rare earth elements.

The uptake behavior of rare earth elements (REEs) under pot conditions using deionized water and a REE fertilizer solution as the culture media as well as the distribution of REEs in rice proteins were studied. The uptake of REEs in rice seeds increased dramatically after a lag period of approximately three days. Roots can accumulate a much higher content of REEs than germs and the resting seeds. The REE content in each water-soluble (albumin) and salt-soluble (globulin) component of the rice seeds accounted for 5-8% and 4-6% of the total REEs, respectively. However, there are less than 1.5% of the total REEs were found in the alcohol-soluble (prolamin) and acetic acid-soluble (glutelin) components. The high performance liquid chromatography (HPLC) in the gel permeation and the reserved-phase were used to monitor changes in the molecular weight distribution changes of the soluble proteins of rice seeds during germination after having been cultured in the same solution for seven days. No changes occurred in the prolamin, while a slight change occurred in the albumin, globulin and glutelin. Fractionation of the albumin of rice seeds cultured in a REE fertilizer solution on the Sephadex G-100 column indicated that REEs, especially Ce, La, Pr and Nd, were associated mainly with biological compounds of a molecular weight between 10,000 and 12,000.

Absorption↗
PubMed2001

Zap-70-independent Ca(2+) mobilization and Erk activation in Jurkat T cells in response to T-cell antigen receptor ligation.

The tyrosine kinase ZAP-70 has been implicated as a critical intermediary between T-cell antigen receptor (TCR) stimulation and Erk activation on the basis of the ability of dominant negative ZAP-70 to inhibit TCR-stimulated Erk activation, and the reported inability of anti-CD3 antibodies to activate Erk in ZAP-70-negative Jurkat cells. However, Erk is activated in T cells receiving a partial agonist signal, despite failing to activate ZAP-70. This discrepancy led us to reanalyze the ZAP-70-negative Jurkat T-cell line P116 for its ability to support Erk activation in response to TCR/CD3 stimulation. Erk was activated by CD3 cross-linking in P116 cells. However, this response required a higher concentration of anti-CD3 antibody and was delayed and transient compared to that in Jurkat T cells. Activation of Raf-1 and MEK-1 was coincident with Erk activation. Remarkably, the time course of Ras activation was comparable in the two cell lines, despite proceeding in the absence of LAT tyrosine phosphorylation in the P116 cells. CD3 stimulation of P116 cells also induced tyrosine phosphorylation of phospholipase C-gamma1 (PLCgamma1) and increased the intracellular Ca(2+) concentration. Protein kinase C (PKC) inhibitors blocked CD3-stimulated Erk activation in P116 cells, while parental Jurkat cells were refractory to PKC inhibition. The physiologic relevance of these signaling events is further supported by the finding of PLCgamma1 tyrosine phosphorylation, Erk activation, and CD69 upregulation in P116 cells on stimulation with superantigen and antigen-presenting cells. These results demonstrate the existence of two pathways leading to TCR-stimulated Erk activation in Jurkat T cells: a ZAP-70-independent pathway requiring PKC and a ZAP-70-dependent pathway that is PKC independent.

CD3 Complex↗
PubMed2001

[Detection of TT virus DNA and sequence analysis of TTV gene in sera from young children in Nanjing].

OBJECTIVE: To investigate the prevalence of TTV genomes in young children from Nanjing and establish the PCR method for detecting TTV DNA. METHODS: A hemi-nested polymerase chain reaction (PCR) assay was established to detect TTV DNA in 110 young children and two PCR-amplified products were sequenced and the data were analyzed with the computer programs. RESULTS: TTV DNA was detected in 14 out of 110 children, and 222 bp sequences of 2 isolates from the children were analyzed. Compared with representative of published isolates, they have the nucleotide identity of 93.7%-98.6% with TX011. CONCLUSIONS: The infection rate of TTV in children is 12.73%, and 2 isolates from children belong to subgroup G1b. Our results indicates that TTV could be transmitted via other pathway besides hematogenous one, and TTV could be carried by healthy population.

Base Sequence↗
PubMed2000

Increased nondisjunction of chromosome 21 with age in human peripheral lymphocytes.

Fluorescence in situ hybridization (FISH) on binucleated cells with chromosome-specific DNA probes provides a convenient way to visualize reciprocal segregation patterns in daughter nuclei, and overcomes most problems related to the artefactual loss or gain of chromosomes that flaw chromosome preparations. In this study, FISH was employed to evaluate age- and sex-effects on spontaneous malsegregation, nondisjunction and loss of chromosome 21 in human lymphocytes after the first division in culture. A total of 68 healthy nonsmokers and nondrinkers of alcohol (37 males and 31 females) were grouped by age as Group I (0-10 years), Group II (20-30 years), Group III (40-50 years) and Group IV (60-70 years), with at least seven subjects per group and sex. FISH with a pericentric chromosome 21 specific DNA probe was carried out on binucleated lymphocytes, cytokinesis-blocked by cytochalasin B (6 microg/ml for 26 h) at 44 h after initiation of cultures. Linear regression analyses demonstrated a significant age-related increase in the frequency of micronuclei without chromosome 21 (MN-21)(r=0.73, p<0.001 in females; r=0.69, p<0.001 in males) in all binucleated cells, with a steeper slope in females (0.1758) than in males (0. 1241). Analysis using the 2x2 chi-square (chi(2)) test on the frequencies of MN-21 showed significant age-related differences in both males and females, except males in Group III and Group IV (p>0. 05). A significant sex-related difference was found only in subjects over 60 years (p<0.05), with females having more MN-21 (12.57 per thousand vs. 8.43 per thousand) than males. Loss of chromosome 21, occurring at mean levels of 0.38 per thousand in all binucleated cells and 0.24 per thousand in binucleated cells containing four FISH signals, was shown not to be age- or sex-related. A positive age-related increase in nondisjunction of chromosome 21 was shown in males (r=0.50, p<0.01), females (r=0.61, p<0.001) and all subjects (r=0.55, p<0.001) by linear regression analysis. An age effect was found only between children and adults (p<0.01 for females, p<0.05

Adolescent↗
PubMed2000

The Drosophila sox gene, fish-hook, is required for postembryonic development.

In Drosophila, the fish-hook (fish) gene encodes a Sox protein essential for embryonic segmentation and nervous system organization. In this study we examined potential functional roles of fish in postembryonic developmental processes, including those involved in adult appendage development. We show here that Fish protein is expressed in discrete patterns in the larval eye-antennal and leg imaginal discs, the central nervous system, the hindgut, and salivary glands. Genetic mosaic studies indicated that fish function is required for the growth or survival of imaginal cells, and the expression of engrailed and wingless. Ectopic expression of Fish protein resulted in severe disruption of adult structures; legs and antennae were truncated and eye formation was suppressed. These morphological defects were correlated with altered expression patterns of the wingless, decapentaplegic, and bric-a-brac genes. Finally, analysis of truncated versions of Fish protein indicated that the HMG domain was sufficient for Fish nuclear localization and that removal of the transcriptional activation domain did not eliminate Fish function. While Sox proteins have been shown to be important for eye and limb formation in vertebrates, these data provide the first evidence for Sox protein functions in appendage development in invertebrates.

Animals↗
PubMed2000

A role for Drosophila Drac1 in neurite outgrowth and synaptogenesis in the giant fiber system.

Recent studies have shown the small GTPases, Rac1, Rho, and CDC42, to have a role in axon guidance. To assess their participation in synapse assembly and function we have expressed various forms of Drac1 in the giant fiber system of Drosophila. Overexpression of wild-type Drac1 in the giant fiber (GF) lead to a disruption in axonal morphology; axons often terminate prematurely in a large swelling in the target area but lack the normal lateral bend where the synapse with the jump motor neuron would normally be found. Electrophysiological assays revealed longer latencies and lowering following frequencies indicating defects in the synapse between the GF and the tergotrochanteral motor neuron (TTMn). Thickened abnormal GF dendrites were also observed in the brain. Overexpression of the dominant-negative form of Drac1, (N17), resulted in axons that produced extra branches in the second thoracic neuromere (T2); however, the synaptic connection to the TTMn was present and functioned normally. Conversely, expression of the constitutively active form, Drac1(V12), resulted in a complete lack of neurite outgrowth and this was also seen with overexpression of Dcdc42(V12). In the absence of a GF, these flies showed no response in the jump (TTM) or flight (DLM) muscles upon brain stimulation. Taken together these results show that the balance of actin polymerization and depolymerization determines local process outgrowth and thereby synapse structure and function.

Action Potentials↗
PubMed2000

Incidence of mosaic cell lines in vivo and malsegregation of chromosome 21 in lymphocytes in vitro of trisomy 21 patients: detection by fluorescence in situ hybridization on binucleated lymphocytes.

In order to detect aneuploidy in interphase human lymphocytes, both in vivo and in vitro, fluorescence in situ hybridization (FISH) was carried out on binucleated cells cytokinesis-blocked by cytochalasin B at the first mitosis after phytohemagglutinin stimulation. A pericentric chromosome-21-specific DNA probe prepared from yeast artificial chromosome clone 881D2 by the polymerase chain reaction was employed. One thousand binucleated cells per individual were scored from cultures from twelve trisomy 21 patients aged 0.01-8.9 years (mean 4.3 years) and 20 normal children of similar age. Of trisomy 21 patients, increased frequencies of disomic cells in vivo (1.690+/-1.070%) and cells containing six signals with nondisjunction (0.822+/-0.554%) were found, compared with those of monosomic 21 cells in vivo (0.265+/-0.130%) and cells containing four signals with nondisjunction in normal children (0.369+/-0.250%; P=0.000 and P=0.000, respectively). These results show that malsegregation of chromosome 21 occurs more often in trisomic 21 cells than in disomic cells from normal children. The frequency of nondisjunction was significantly higher than the loss of chromosome 21 in both cultured trisomic (0.822+/-0.554% vs 0.043+/-0.049%, P=0.000) and disomic (0.369+/-0.250% vs 0.010+/-0.30%, P=0.000) cells. Comparisons of in vivo and in vitro data on aneuploidy indicate that a cell selection mechanism may exist in vivo. All these results show that FISH, with a chromosome-specific probe, on binucleated lymphocytes is a powerful tool for simultaneously detecting mosaic cell lines in vivo and malsegregation (loss and nondisjunction) of a corresponding chromosome in vitro in the same cell population.

Case-Control Studies↗
PubMed2000

Self-organized criticality in a computer network model

We study the collective behavior of computer network nodes by using a cellular automaton model. The results show that when the load of network is constant, the throughputs and buffer contents of nodes are power-law distributed in both space and time. Also the feature of 1/f noise appears in the power spectrum of the change of the number of nodes that bear a fixed part of the system load. It can be seen as yet another example of self-organized criticality. Power-law decay in the distribution of buffer contents implies that heavy network congestion occurs with small probability. The temporal power-law distribution for throughput might be a reasonable explanation for the observed self-similarity in computer network traffic.

Journal Article↗
PubMed2000

Deficiency of PTEN in Jurkat T cells causes constitutive localization of Itk to the plasma membrane and hyperresponsiveness to CD3 stimulation.

Pleckstrin homology (PH) domain binding to D3-phosphorylated phosphatidylinositides (PI) provides a reversible means of recruiting proteins to the plasma membrane, with the resultant change in subcellular localization playing a key role in the activation of multiple intracellular signaling pathways. Previously we found that the T-cell-specific PH domain-containing kinase Itk is constitutively membrane associated in Jurkat T cells. This distribution was unexpected given that the closely related B-cell kinase, Btk, is almost exclusively cytosolic. In addition to constitutive membrane association of Itk, unstimulated JTAg T cells also exhibited constitutive phosphorylation of Akt on Ser-473, an indication of elevated basal levels of the phosphatidylinositol 3-kinase (PI3K) products PI-3,4-P(2) and PI-3,4,5-P(3) in the plasma membrane. Here we describe a defect in expression of the D3 phosphoinositide phosphatase, PTEN, in Jurkat and JTAg T cells that leads to unregulated PH domain interactions with the plasma membrane. Inhibition of D3 phosphorylation by PI3K inhibitors, or by expression of PTEN, blocked constitutive phosphorylation of Akt on Ser-473 and caused Itk to redistribute to the cytosol. The PTEN-deficient cells were also hyperresponsive to T-cell receptor (TCR) stimulation, as measured by Itk kinase activity, tyrosine phosphorylation of phospholipase C-gamma1, and activation of Erk compared to those in PTEN-replete cells. These data support the idea that PH domain-mediated association with the plasma membrane is required for Itk activation, provide evidence for a negative regulatory role of PTEN in TCR stimulation, and suggest that signaling models based on results from Jurkat T-cell lines may underestimate the role of PI3K in TCR signaling.

Animals↗
PubMed2000

[Correlation of inactivation of CDKN2/p16 gene by methylation and lung cancers].

OBJECTIVE: To study the relationship between methylation state of CDKN2/p16 gene and lung cancers. METHODS: Genomic DNA was digested with methylation-sensitive enzymes and Southern blotting was used to analyses the methylation state of CDKN2/p16 gene in 89 cases of lung cancer. RESULTS: Twenty-one out of 89 lung cancers(23.6%) shown methylation of CDKN2/p16 gene, and 17 of them occurred in 42 P16 negative cases with a rate of 40.5%(17/42). CONCLUSION: Methylation of CDKN2/p16 gene 5' CpG island is probably an important mechanism of gene inactivation; it may take part in the onset and process of lung cancer.

Adult↗
PubMed2000

Detection of mosaic chromosome 21 aneuploidy in vivo with CB-FISH method.

OBJECTIVE: To explore the mechanism by which the ratio of mosaic diploid cells in vivo increased in trisomy 21 cases. METHODS: Fluorescence in situ hybridization (FISH) with pericentric DNA probes specific to human chromosome 21 to the binucleated cells cytokinesis- blocked by cytochalasin-B(CB), namely, CB-FISH method. RESULTS: The mean frequency of diploid cells (1.69%+/-1.07%) was significantly higher than that of tetrasomy 21 cells(0.042%+/-0.064%)(P=0.000) in 12 trisomy 21 cases aged 0-9 years (mean 4 years) in vivo; whereas the incidence of spontaneous monosomy 21 cells (0. 265%+/-0.13%) was close to the frequency of trisomy 21 cells (0. 225%+/-0.184%) in 20 control individuals aged 0-10 years (mean 5 years). The frequency of cells undergone chromosome 21 nondisjunction was 0.822%+/-0.554% in cultured trisomy 21 cells, which was significantly higher than that in the cultured diploid cells (0.369%+/-0.25%). However, the rates of cells with chromosome 21 loss in the two cultured cell lines (0.01%+/-0.03% vs 0.043%+/-0.049%) were not significantly different. In both cultured cell lines, chromosome 21 nondisjunction occurred more frequently than loss. CONCLUSION: The increased frequency of diploidies in trisomy 21 cases may result from the accumulation of diploid cells originating from malsegregation of chromosome 21 in trisomic cells rather than the increase of chromosome 21 loss.

Aneuploidy↗