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Biomedical subjects

C P Chang

Publications and source records attributed to C P Chang.

At least 37 records · Page 2Linked to original sources
PubMed1999

Mutation analysis of the PTEN/MMAC1 gene in cancers of the digestive tract.

The 10q23.3 gene PTEN (phosphatase and Tensin homologue deleted on chromosome 10) or MMAC1 (mutated in multiple advanced cancers 1) was recently reported to undergo frequent mutation, including mutations and deletions in multiple advanced cancers. This study showed that the aberrant transcripts of this gene are frequently found in cancers of the digestive tract, paired non-cancerous tissues and normal peripheral mononuclear cells. Sequence analysis of the aberrant transcripts revealed three types of deletions: (i) a deletion junction with a splicing-like donor or acceptor sequence; (ii) several-base homology near or between the donor acceptor site at the deletion junction; and (iii) deletion with insertion. From these results, it is suggested that aberrant transcripts of PTEN/MMAC1 found by nested reverse transcription-polymerase chain reaction are a common (or natural) phenomenon unrelated to oncogenesis. The mechanism producing these aberrant transcripts needs further investigation. Using single-strand conformation polymorphism and direct sequencing to analyse for small base changes of the genomic DNA of the PTEN/MMAC1 gene revealed no point mutations or small base changes.

Colonic Neoplasms↗
PubMed1999

Analysis of TSG101 tumour susceptibility gene transcripts in cervical and endometrial cancers.

Carcinoma of the uterine cervix is a common malignancy among women that has been found to show loss of heterozygosity in the chromosome 11p. Recent studies have localized the TSG101 gene in this region, and also demonstrated a high frequency of abnormalities of this gene in human breast cancer. To determine the role of the TSG101 gene in the carcinogenesis of cervical and uterine carcinoma, 19 cases of cervical carcinoma and five cases of endometrial carcinoma, as well as nearby non-cancerous tissue from the same patients, and 16 blood samples from healthy persons as normal control were analysed by Southern blot analysis of genomic DNA, reverse transcription of the TSG101 mRNA followed by PCR amplification and sequencing of the products. We found that abnormal transcripts of the TSG101 gene were common both in cancerous or non-cancerous tissues of the uterus and cervix and in normal peripheral mononuclear cells. There was no genomic deletion or rearrangement in spite of the presence of abnormal transcripts, and no definite relationship between the abnormal transcripts and HPV infection was found. Although the frequency of abnormal transcripts was higher in cancerous than in non-cancerous tissue, normal peripheral mononuclear cells also had abnormal transcripts. Given these findings, the role of the TSG101 gene as a tumour-suppressor gene should be re-evaluated. Because some aberrant transcripts could be found at the first PCR reaction, we suggest that the aberrant transcripts might be the result of imperfect minor splicesome products.

Adult↗
PubMed1999

Acute tumour lysis syndrome: a case in AL amyloidosis.

Tumour lysis syndrome (TLS) in plasma cell dyscrasias is extremely rare. TLS has been described in eight cases of multiple myeloma undergoing high-dose therapy with autologous stem cell transplant (ASCT). Recently, clinical trials of intensive chemotherapy followed by autologous or allogeneic stem cell support has been shown to offer potential benefit in AL (amyloid light-chain) amyloidosis. TLS in primary AL amyloidosis in this setting has not been previously reported. We report a case of TLS in a patient with AL amyloidosis which developed after high-dose melphalan chemotherapy supported by ASCT.

Acute Disease↗
PubMed1999

Differential distribution of apolipoprotein E in young and aged spontaneously hypertensive and stroke-prone rats.

OBJECTIVES: To evaluate whether the differential distribution of apolipoprotein E among lipoprotein fractions and hepatic expression of the apolipoprotein E gene are causal factors in the regulation of lipid metabolism and physiological functions in young and aged spontaneously hypertensive and stroke-prone rats. DESIGN AND METHODS: Biochemical analyses were performed on serum and hepatic specimens from young (2-month-old) and aged (8-month-old) spontaneously hypertensive rats, stroke-prone spontaneously hypertensive rats and normotensive Wistar-Kyoto rats. Levels of apolipoprotein E among various lipoprotein fractions were determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Liver concentrations of apolipoprotein E mRNA were analyzed by Northern blotting and relative levels of apolipoprotein E mRNA in different strains of rats were compared. Statistical analysis was performed by measuring correlations between hepatic apolipoprotein E mRNA levels and biological parameters. RESULTS: Levels of apolipoprotein E in high-density and low-density lipoproteins were significantly lower in hypertensive rats than in age-matched normotensive Wistar- Kyoto rats. Although there was a significant increase in high-density lipoprotein apolipoprotein E contents in all aged animals, the elevation in aged hypertensive rats was much less than that in aged normotensive rats. Levels of apolipoprotein E in the very-low-density lipoprotein fraction were diminished in young stroke-prone rats but were remarkably high in aged rats. Steady-state levels of apolipoprotein E mRNA increased with age in all strains of rats, whereas aged hypertensive rats exhibited lower apolipoprotein E mRNA levels than aged normotensive rats. CONCLUSIONS: The distribution of apolipoprotein E among various lipoprotein fractions was dramatically altered with age, and the alteration varied among different strains of rats. The differential distribution of apolipoprotein E in young and aged spontaneously hypertensive and stroke-prone rats suggests that apolipoprotein E could be a causal factor that disturbs the homeostasis of lipids and lipoproteins and perturbs physiological functions in hypertensive rats.

Aging↗
PubMed1999

Different patterns of serum chromogranin A in patients with prostate cancer with and without undergoing hormonal therapy.

Elevated serum chromogranin A (CgA) levels have been detected in patients with prostate cancer who have developed resistance to hormonal therapy. We would like to reexamine these cases by using serial specimens to determine whether such elevated levels are also detectable in prostate cancer patients not undergoing hormonal therapy. Serum CgA was measured in both random and serial specimens from prostate cancer patients with and without undergoing hormonal therapy. We found that serum CgA levels became elevated much earlier than did the levels of serum PSA in approximately one-third of prostate cancer patients developing resistance to hormonal therapy. On the other hand, serum CgA levels became elevated at later, more advanced stages of the disease in patients not undergoing hormonal therapy. Elevated serum CgA levels were usually detected in specimens containing highly elevated PSA. The early rise of serum CgA levels provides an early signal allowing a change of therapy to be made before the disease progresses to a fatal stage. Drugs targeting neuroendocrine cells should be considered for prostate cancer patients with elevated serum CgA levels.

Antineoplastic Agents, Hormonal↗
PubMed1999

Development of a microplate assay for serum chromogranin A (CgA): establishment of normal reference values and detection of elevated CgA in malignant diseases.

Chromogranin A (CgA), a marker for neuroendocrine cells, is associated with poor prognosis when detected by immunohistochemical technique in prostate tumors. We have developed an ELISA on microplates for serum CgA and established the normal reference range. We also attempted to find out whether elevated serum CgA levels could be found in patients with various malignant diseases. Because of non-Gaussian distribution, both medians and 97.5 percentiles of serum CgA levels for men and women of four different age groups were determined. For women, the median and 97.5 percentiles are 20.7 and 63.9 ng/mL for ages 20 to 50, and 32 and 93.8 for 50 to 80 years of age, respectively; for men, they are 27.9 and 78.4 ng/mL for ages 18 to 40 and 41.6 and 92 for 40 to 80 years old, respectively. Elevated serum concentrations of CgA were detectable in patients with prostate cancer not undergoing hormonal treatment, and in patients with various malignant diseases including nonendocrine carcinomas. Most elevated serum CgA levels were associated with sera containing highly elevated serum tumor markers. Drugs targeting neuroendocrine cells should be administered for cancer patients with elevated serum CgA levels.

Adult↗
PubMed1998

A novel protein complex that interacts with the vitamin D3 receptor in a ligand-dependent manner and enhances VDR transactivation in a cell-free system.

Nuclear receptors transduce hormonal signals by binding directly to DNA target sites in promoters and modulating the transcription of linked genes. Receptor-mediated transactivation appears to be potentiated in response to ligand by a number of coactivators that may provide key interactions with components of the transcription preinitiation complex and/or alter chromatin structure. Here, we use the vitamin D3 receptor ligand-binding domain (VDR LBD) as an affinity matrix to identify components of a transcriptionally active nuclear extract that interact with VDR in response to ligand. We describe the purification of a complex of at least 10 VDR interacting proteins (DRIPs) ranging from 65 to 250 kD that associate with the receptor in a strictly 1,25-dihydroxyvitamin D3-dependent manner. These proteins also appear to interact with other, but not all, nuclear receptors, such as the thyroid hormone receptor. The DRIPs are distinct from known nuclear receptor coactivators, although like these coactivators, their interaction also requires the AF-2 transactivation motif of VDR. In addition, the DRIP complex contains histone acetyltransferase activity, indicating that at least one or more of the DRIPs may function at the level of nucleosomal modification. However, we show that the DRIPs selectively enhance the transcriptional activity of VDR on a naked DNA template utilizing a cell-free, ligand-dependent transcription assay. Moreover, this activity can be specifically depleted from the extract by liganded, but not unliganded, VDR-LBD. Overexpression of DRIP100 in vivo resulted in a strong squelching of VDR transactivation, suggesting the sequestration of other limiting factors, including components of the DRIP complex. These results demonstrate the existence of a new complex of novel functional nuclear receptor coactivators.

Acetyltransferases↗
PubMed1998

Analysis of FHIT transcripts in cervical and endometrial cancers.

Carcinoma of the uterine cervix is a common malignancy, and many affected women, have been found to exhibit loss of heterozygosity (LOH) in the chromosome 3p region. Recent studies have localized the FHIT (fragile histidine triad) gene in this region and also demonstrated a high frequency of abnormalities of this gene in various cancers. To determine the role of the FHIT gene in cervical and uterine carcinomas, 16 cases of cervical carcinoma and 7 cases of endometrial carcinoma, as well as nearby non-cancerous tissues in these patients, were analyzed by reverse transcription of the FHIT mRNA followed by polymerase chain reaction amplification and sequencing of the products. In this study, 13 of 16 cervical cancers and 4 of 7 endometrial cancers displayed abnormal FHIT transcripts, including a lack of 2 or more exons of the FHIT gene, the insertion of several bases in the deletion junctions, and a 282 bp deletion from cDNA 171 to 452, resulting in a frameshift. Moreover, 5 of 16 matched non-cancerous tissues from the cervical cancer patients and 4 of 7 non-cancerous tissues from endometrial cancer patients also showed the presence of abnormal transcripts lacking 3 or more exons of the FHIT gene. Only 1 of 23 paired samples exhibited LOH. Our results suggest that the abnormal transcript of the FHIT gene is common in both normal and tumor tissues of the uterus and cervix. We also checked for HPV infection in these samples and found no definite relationship between the abnormal transcript and human papillomavirus infection.

Acid Anhydride Hydrolases↗
PubMed1998

Rapid detection of a recombinant hotspot associated with Charcot-Marie-Tooth disease type IA duplication by a PCR-based DNA test.

A 1.5-Mb duplication on chromosome 17p11.2-p12 (CMT1A duplication) caused by a misalignment of the CMT1A repeat sequences (CMT1A-REPs) is associated with Charcot-Marie-Tooth disease type 1A (CMT1A). A hotspot of crossover breakpoints located in a 3.2-kb region of the CMT1A-REPs accounts for three-quarters of the rearrangements in CMT1A patients. We developed a PCR-based diagnostic method to detect a recombination hotspot associated with the CMT1A duplication. Thirty-one CMT1A Chinese patients from different families and 50 healthy people over 65 years of age were studied. Twenty-seven of the 31 cases demonstrated the 3.2-kb hotspot crossover, of which there were two subgroups. The type 1 crossover breakpoint was located at the distal CMT1A-REP around the PmeI site, and accounted for 24 of the 27 cases with a 3.2-kb hotspot crossover in CMT1A duplication patients. The type 2 crossover breakpoint was located at the distal CMT1A-REP around the base 3625 region, accounting for 3 of the 27 cases. The results correlated very well with the results of Southern transfer analysis. This study has a potentially important role in the diagnosis of CMT1A disease.

Charcot-Marie-Tooth Disease↗
PubMed1997

Chimeric oncoprotein E2a-Pbx1 induces apoptosis of hematopoietic cells by a p53-independent mechanism that is suppressed by Bcl-2.

The chimeric oncoprotein E2a-Pbx1 results from fusion of the E2A and PBX1 genes following t(1;19) chromosomal translocations in B cell precursor acute leukemias. Experimentally B cell progenitors do not tolerate constitutive expression of E2a-Pbx1 which contrasts with transformation of several other cell types following its stable expression both in vitro and in vivo. To further investigate the effects of E2a-Pbx1 on the B cell progenitors, we conditionally expressed E2a-Pbx1 under control of a metal response element in hematopoietic precursor cell lines in vitro. Inducible expression of E2a-Pbx1 resulted in cell death with the morphologic and molecular features of apoptosis. A structure-function analysis demonstrated that induction of apoptosis was not a dominant-negative effect of the E2a moiety but, rather, required the DNA-binding homeodomain of Pbx1. E2a-Pbx1-induced apoptosis proceeded through a BCL2-responsive checkpoint eventuating in PARP inactivation but did require p53. Constitutive expression of E2a-Pbx1 did not induce apoptosis or continued cycling of Rat-1 fibroblasts in low serum conditions. These studies demonstrate that E2a-Pbx1 initiates programmed cell death of hematopoietic precursers by a mechanism that requires its chimeric transcriptional properties, but, unlike other nuclear oncoproteins, is independent of p53.

Animals↗
PubMed1997

Valtrac-secured intracolonic bypass device: an experimental study.

PURPOSE: The intracolonic bypass tube has been used both experimentally and clinically to protect the anastomotic site. A newly designed intracolonic bypass, the Valtrac-secured intracolonic bypass, which consisted of a biofragmentable anastomosis ring (BAR) and was connected with a soft, thin vinyl tube, was used in the colon approximately 5 to 10 cm proximal to the anastomotic site. The distal end of the vinyl tube is passed through the colonic anastomosis to the anus to bypass the fecal stream. METHODS: Eighteen piglets were divided into three groups of six each. Group A piglets underwent colon resection and rough anastomosis with large gaps between sutures, followed by Valtrac-secured intracolonic bypass. Group B piglets underwent the same procedures, but a colonic outlet obstruction also was done with pursestring sutures tied over the anus. Group C piglets underwent colon resection and rough anastomosis, but no intracolonic bypass tube was inserted (as in the control group). RESULTS: All Group A and Group B piglets survived. Passage of the BARs occurred approximately two weeks later. As the barium enema passed through the bypass tube, it showed a patent BAR-secured tube and intact anastomosis with no leakage. In Group C, anastomotic leakage occurred in four of six piglets, three of which died. Barium enema showed leakage at the anastomotic site. CONCLUSIONS: In the animal model we used, our new intracolonic bypass device proved to be a simple, safe, reliable means of protecting the anastomotic site and, thereby, eliminated the need for a diverting colostomy. Still we need further steps to test its potential in clinical use.

Anastomosis, Surgical↗
PubMed1997

Molecular analysis of survival motor neuron (SMN) and neuronal apoptosis inhibitory protein (NAIP) genes of spinal muscular atrophy patients and their parents.

We have assayed deletions of two candidate genes for spinal muscular atrophy (SMA), the survival motor neuron (SMN) and neuronal apoptosis inhibitory protein (NAIP) genes, in 101 patients from 86 Chinese SMA families. Deletions of exons 7 and 8 of the telomeric SMN gene were detected in 100%, 78.6%, 96.6%, and 16.7%, in type I, II, III, and adult-onset SMA patients, respectively. Deletion of exon 7 only was found in eight type II and one type III patient. One type II patient did not have a deletion of either exon 7 or 8. The prevalence of deletions of exons 5 and 6 of the NAIP gene were 22.5% and 2.4% in type I and II SMA patients, respectively. We also examined four polymorphisms of SMN genes and found that there were only two, SMN-2 and CBCD541-2, in Chinese subjects. In our study, analysis of the ratio of the telomeric to centromeric portion (T/C ratio) of the SMN gene after enzyme digestion was performed to differentiate carriers, normals, and SMA patients. We found the T/C ratio of exon 7 of the SMN gene differed significantly among the three groups, and may be used for carrier analysis. An asymptomatic individual with homozygous deletion of exons 7 and 8 of the SMN gene showed no difference in microsatellite markers in the SMA-related 5q11.2-5q13.3. In conclusion, SMN deletion in clinically presumed child-onset SMA should be considered as confirmation of the diagnosis. However, adult-onset SMA, a heterogeneous disease with phenotypical similarities to child-onset SMA, may be caused by SMN or other gene(s).

Adult↗
PubMed1997

Differential expression of the alpha and beta subunits of the large-conductance calcium-activated potassium channel: implication for channel diversity.

In addition to the large alpha subunits that conduct selective ion currents, many native voltage-gated ion channels contain associated proteins which modulate the channel activity. Recently, a beta subunit of the large-conductance calcium-activated K+ (BK) channel has been cloned and functionally characterized. In this report, we studied the tissue distribution of the alpha and beta subunits of rat BK channels by nuclease protection analyses and in situ hybridization. BK alpha mRNA is widely distributed but is especially enriched in the brain. In the adult brain, BK alpha expression is robust and widespread throughout all areas of the neo-, olfactory and hippocampal cortices, habenula and cerebellum. Other prominent sites of BK alpha expression include thalamus and amygdala. In marked contrast to the expression pattern of BK alpha mRNA, the expression of BK beta mRNA is relatively low and preferentially in the periphery. In rat brains, BK beta mRNA occurs only in a few discrete populations of neurons that also express BK alpha messages. These results indicate that the major type of BK channels in the brain, unlike the alpha beta channel type in aortic and tracheal smooth muscle, is devoid of the beta subunit. These observations provide a structural basis for the BK channel diversity observed in a variety of tissues.

Amino Acid Sequence↗
PubMed1997

The Hox cooperativity motif of the chimeric oncoprotein E2a-Pbx1 is necessary and sufficient for oncogenesis.

E2a-Pbx1 chimeric oncoproteins result from fusion of the E2A and PBX1 genes at the sites of t(1;19) chromosomal translocations in a subset acute lymphoblastic leukemias. Experimentally, E2a-Pbx1 transforms a variety of cell types, including fibroblasts, myeloid progenitors, and lymphoblasts. Structure-function studies have shown that contributions from both E2a and Pbx1 are necessary for oncogenesis, but the Pbx1 homeodomain is dispensable and the required portion of Pbx1 has not been delineated. In this study, we used deletional and site-directed mutagenesis to identify portions of Pbx1 necessary for oncogenic and transcriptional activities of E2a-Pbx1. These studies defined a motif (named the Hox cooperativity motif [HCM]) carboxy terminal to the Pbx homeodomain that is required for cooperative DNA binding, cellular transcriptional activity, and the oncogenic potential of E2a-Pbx1. The HCM is highly conserved throughout the Pbx/exd subfamily of divergent homeodomain proteins and functions in DNA-binding assays as a potential contact site for Hox dimerization. E2a-Pbx1 proteins with interstitial deletion or single-point mutations in the HCM could neither activate transcription in cellular assays nor transform NIH 3T3 cells. An E2a-Pbx1 mutant containing 50 amino acids of Pbx1b spanning the HCM but lacking the homeodomain was capable of inducing fibroblast transformation. Thus, the HCM is a necessary and sufficient contribution of Pbx1 for oncogenesis induced by E2a-Pbx1 and accounts for its homeodomain-independent transforming properties. Since subtle alterations of the Pbx HCM result in complete abrogation of transforming activity whereas the homeodomain is entirely dispensable, we conclude that interactions mediated by the HCM are more important for transformation by E2a-Pbx1 than interactions with cognate Pbx DNA sites.

3T3 Cells↗
PubMed1997

Meis proteins are major in vivo DNA binding partners for wild-type but not chimeric Pbx proteins.

The Pbx1 and Meis1 proto-oncogenes code for divergent homeodomain proteins that are targets for oncogenic mutations in human and murine leukemias, respectively, and implicated by genetic analyses to functionally collaborate with Hox proteins during embryonic development and/or oncogenesis. Although Pbx proteins have been shown to dimerize with Hox proteins and modulate their DNA binding properties in vitro, the biochemical compositions of endogenous Pbx-containing complexes have not been determined. In the present study, we demonstrate that Pbx and Meis proteins form abundant complexes that comprise a major Pbx-containing DNA binding activity in nuclear extracts of cultured cells and mouse embryos. Pbx1 and Meis1 dimerize in solution and cooperatively bind bipartite DNA sequences consisting of directly adjacent Pbx and Meis half sites. Pbx1-Meis1 heterodimers display distinctive DNA binding specificities and cross-bind to a subset of Pbx-Hox sites, including those previously implicated as response elements for the execution of Pbx-dependent Hox programs in vivo. Chimeric oncoprotein E2a-Pbx1 is unable to bind DNA with Meis1, due to the deletion of amino-terminal Pbx1 sequences following fusion with E2a. We conclude that Meis proteins are preferred in vivo DNA binding partners for wild-type Pbx1, a relationship that is circumvented by its oncogenic counterpart E2a-Pbx1.

Adenovirus E2 Proteins↗
PubMed1997

Multiplex mutagenically separated PCR: diagnosis of beta-thalassemia and hemoglobin variants.

A rapid and simple method, termed multiplex mutagenically separated PCR (MS-PCR), was developed to detect several molecular defects in the hemoglobin gene in one PCR. This technique, in which different-size allele-specific primers were used, specifically amplified both normal and mutant alleles of the globin gene in the same reaction. Subsequent gel electrophoresis showed at least one of the two allelic products at the same locus or two of the several allelic products of different loci and provided a within-assay quality control for the exclusion of false-negative results. In our study, the four most common beta-thalassemia mutations, together with four other common hemoglobin variants in Chinese, were tested. Using multiplex MS-PCR 6 to 12 primers were added simultaneously into one reaction tube to identify one to four mutations. Not only is this multiplex MS-PCR method reliable and non-isotopic, the results can be obtained in less than one working day.

Alleles↗
PubMed1997

Identification and differentiation of bear bile used in medicinal products in Taiwan.

One hundred eighty-three suspect bear bile used in medicinal products, collected in Taiwan as gall bladders or dried powder forms, were analyzed using FTIR, HPTLC, and HPLC techniques to identify whether they are indeed bear bile. Those confirmed were further examined to determine whether the observed analytical parameters can be reliably used for source inference, i.e., differentiating products among North American black bear, farmed Asiatic black bear, polar bear, etc. Our data suggested that North American and polar bears contain a higher concentration of TC (relative to TUDC and TCDC), whereas the relative concentration of TC in Asiatic bears (wild or farmed) is much lower. Thus, the relative concentration of TC can potentially be used for differentiating Asiatic bear bile from North American and polar bear products, but it cannot be used for the differentiation of wild and farmed bear bile as suggested in an earlier report by Espinoza et al. The origin of the 183 samples analyzed were found to be as follows: 118 (64%), bile salts, or gall bladders were of domestic pig; 56 (31%), bile products of Asiatic bear; 4 (2.2%), Asiatic bear mixed with pig bile salts; 3 (1.6%) goat gall bladders; 1 (0.55%) water buffalo bile salts; and 1 (0.55%), pig bile salts mixed with water buffalo bile salts.

Animals↗