Biomedical subjects

T Y Hsu

Publications and source records attributed to T Y Hsu.

At least 73 records · Page 4Linked to original sources

PubMed1994

Analysis of integrated hepatitis B virus DNA and flanking cellular sequences in the hepatocellular carcinoma cell line HCC36.

A hepatocellular carcinoma cell line, HCC36, was established from an adult HBV carrier in Taiwan. From Southern blot analysis, there were at least four sites of integration of HBV DNA, and no viral replicative intermediates were detected. A genomic library was constructed from HCC36 DNA, and two phage clones, designated lambda 36A and lambda 36B, were shown to contain HBV DNA and flanking cellular sequences. In lambda 36A, HBV DNA sequences were quite conserved, and 7.4% base variation was detected. The viral sequences in lambda 36A and lambda 36B differed in only four bases, in addition to the microdeletion and -insertion observed in lambda 36B. The flanking cellular sequences identified in lambda 36A were human Alu sequences and in lambda 36B satellite sequences.

Base Sequence↗

PubMed1994

Abdominal pregnancy--report of two cases and review of literature.

Ectopic pregnancy is one of the major complications of pregnancy. The abdominal pregnancy is a comparatively rare type of ectopic gestation with a high fetal and maternal morbidity and mortality. Although abdominal pregnancy had been well documented, it still remains a serious dilemma for most clinicians in the contemporary obstetric practice because of the difficulties in early diagnosis and proper management. We reported two cases of early abdominal pregnancy with accurate preoperative diagnosis followed by immediate surgical intervention and total removal of the placental tissue. The important concepts of management with this uncommon but ominous condition are discussed, including the predisposing risk factors, clinical features, diagnostic difficulties, appropriate surgery and management of the placenta.

Adult↗

PubMed1994

Liver-specific expression and high oncogenic efficiency of a c-myc transgene activated by woodchuck hepatitis virus insertion.

The high oncogenic efficiency of woodchuck hepatitis virus (WHV) has been correlated with the ability of this virus to provoke insertional activation of myc family genes. To assess the impact of viral integration on liver cell transformation, we have generated transgenic mice carrying the mutated c-myc gene and adjacent viral DNA from a woodchuck tumor, in original configuration. Virtually all mice from two different strains developed hepatocellular carcinoma with a mean latency period of 8-12 months. The c-myc transgene was expressed transiently in neonatal livers, and re-expressed at preneoplastic and neoplastic stages in adult livers. Woodchuck c-myc mRNA driven by the normal P1 and P2 promoters and WHV-specific transcripts encoding viral surface antigens were produced in a strictly co-regulated fashion during development and tumorigenesis, indicating a predominant regulatory influence of the viral enhancer. Furthermore, the activity of the viral enhancer in response to various biological stimuli was apparently modulated by glucose uptake and glucagon/insulin balance in differentiated hepatocytes. In this model, a viral integration event selected from a naturally occurring tumor proved to be determinant for induction of hepatocarcinogenesis, although enforced, liver-specific expression of c-myc was limited to a particular developmental stage.

Animals↗

PubMed1994

Postmortem chorionic villus sampling for cytogenetic evaluation of early fetal loss: a simple reliable method.

BACKGROUND: Early fetal losses associated with chromosome abnormalities produce most spontaneous abortions. It is not feasible to conduct cytogenetic studies with tissue culture postmortem because of tissue maceration and autolysis. METHODS: A total of 30 specimens from early fetal losses were karyotyped for chromosomal study. Success rate of the culture method is evaluated and compared. RESULTS: The abnormal karyotype rate was 33.3%. It was found that chorionic villus tissues had a higher culture rate than tests of fetal skin, amnion or amniocyte by early amniocentesis. CONCLUSIONS: Postmortem chorionic villus sampling at the time of diagnosis of early fetal death appears to offer the best, most simple and reliable method for obtaining a cytogenetic result, and also a good method for training residents in the chorionic villus sampling technique.

Adult↗

PubMed1994

Stability of HBV DNA in cell lines and nude mouse-passaged tissues derived from human hepatocellular carcinoma.

Human hepatitis B virus (HBV) infection has been closely linked to the occurrence of hepatocellular carcinoma (HCC). Hepatoma cell lines and nude mouse-passaged hepatoma tissues were used in this report to study the HBV DNA status in these cells after passage. DNA was extracted from seven hepatoma cell lines and three nude mouse passaged HCC lines. Southern blot hybridization technique was performed with either cloned HBV whole genome or subgenomic DNA fragments as probes to analyze the presence of HBV DNA. Integration of HBV DNA fragments was detected in one mouse passaged tissue, R. Hybridization with HBV subgenomic DNA revealed that there were some DNA rearrangements of the integrated HBV DNA in R. However, the integrated HBV DNA could not be detected in the cell line derived from R after in vitro cultivation for 2 years. Both episomal form and integrated HBV DNA were detected in a cell line NTU-h3. Episomal form HBV DNA ih NTU-h3 changed after several passages. HBV DNA in NTU-h3 was unstable after in vitro cultivation. Therefore, we concluded that the presence of HBV DNA might not be essential for the maintenance of the tumorigenicity of hepatoma and the nude mouse system was more stable for maintaining HBV DNA in HCC.

Animals↗

PubMed1993

Establishment and characterization of an HTLV-I cell line from a Taiwanese patient with HTLV-I-associated myelopathy.

We describe a Taiwanese woman with chronic progressive myelopathy, in whom Western blot analysis of the serum and cerebrospinal fluid (CSF) displayed positive reactions to human T-lymphotropic virus type I (HTLV-I) proteins, p19, p24, p28, p36, gp46 and p53. HTLV-I proviral genomes were detected in the peripheral blood mononuclear cells (PBMC) and CSF cells by nested polymerase chain reaction and Southern blot hybridization. HTLV-I was successfully isolated from PBMC stimulated with interleukin-2 (IL-2). The established cell line, named THAM-1, was an IL-2-independent T-cell line with CD2+, CD3+, CD4+, CD25+ and HLA-DR+. Retrovirus particles with type C morphology were observed in the THAM-1 cells by electron microscopy, and HTLV-I-related antigens were also demonstrated by immunocytochemical staining and Western blot assay. Southern blot analysis revealed that HTLV-I proviral genomes were integrated into the THAM-1 cellular DNA. In Northern blot analysis, two extra-species of RNA were detected in addition to three typical viral transcripts. For the first time, an HTLV-I-producing T cell line was established from a patient with HTLV-I-associated myelopathy in Taiwan, an HTLV-I non-endemic area.

Adult↗

PubMed1993

Use of bacterially-expressed antigen for detection of antibodies to the EBV-specific deoxyribonuclease in sera from patients with nasopharyngeal carcinoma.

A cDNA clone, BG9, corresponding to the open reading frame BGLF5 of Epstein-Barr virus (EBV) DNase was inserted into an E. coli expression vector, pET3a, to generate a recombinant plasmid, pDNase 5. High level of expression of a DNase activity was detected in the E. coli transformed with pDNase 5 following induction with IPTG. The enzyme activity was purified using DEAE-cellulose, phosphocellulose and DNA-cellulose column chromatography. The purified protein appeared to be nearly homogeneous in SDS-PAGE using Coomassie blue staining. The requirement for divalent cations and optimum pH as well as inhibitory concentrations of ionic strength and polyamines for the purified enzyme activity were determined and seemed to be very similar to those of the enzyme activity purified from an EBV producing lymphoblastoid cell line. Using the purified enzyme as an antigen and anti-IgA as the secondary antibody, 82% (64/78) and 91% (71/78) of sera from patients with nasopharyngeal carcinoma (NPC) were shown to be positive by dot immunobinding assay and ELISA, respectively. The results suggest that purified E. coli expressed EBV DNase may be useful for preparing specific test for large scale screening of patients with NPC.

Antibodies, Viral↗

PubMed1993

Analysis of a DNA polymorphic region in the gtfB and gtfC genes of Streptococcus mutans.

We have previously demonstrated the existence of DNA polymorphisms at the 5' coding regions of the gtfB and gtfC genes specifying the streptococcal glucosyltransferases (J.S. Chia, T.Y. Hsu, L.J. Teng, J.Y. Chen, L.J. Hahn, and C.S. Yang, Infect. Immun. 59:1656-1660, 1991). DNA sequence analysis by polymerase chain reaction and direct sequencing revealed that while several nucleotide changes were identified, accounting for the polymorphisms, the amino acids which they code for remain unchanged. The polymorphic region is located in a highly conserved amino terminus of the glucosyltransferases. A peptide of 19 amino acids from this region reversed the inhibiting activity of an antiserum raised against the proteins coded for by the gtfB and gtfC genes. The results suggest that the polymorphic region, varying in DNA but not in amino acid sequences, might specify some biological function.

Amino Acid Sequence↗

PubMed1993

Analysis of Epstein-Barr virus BamHI F DNA fragment in nude mouse-passaged nasopharyngeal carcinoma tissues.

Two nude mouse-passaged nasopharyngeal carcinoma (NPC) tissues, NPC-295 and NPC-306, were different in ability to transform cord blood lymphocytes and contained Epstein-Barr virus (EBV) genomes with difference in BamHI F DNA fragment. Four clones containing DNA sequences of the BamHI F fragment (54,853-62,249) were obtained from genomic libraries of NPC-295 and NPC-306 and their partial restriction enzyme maps and sequences were determined. The restriction enzyme maps of EBV DNA at the BamHI F fragment in NPC-295 and NPC-306 appeared to be similar to that of EBV B95-8 strain. However, 8 nucleotide differences were revealed between NPC-295 and the B95-8 strain when 566 nucleotides (55,405-55,970) were compared, and 15 out of 677 nucleotides analyzed (55,410-56,086) were different between NPC-306 and B95-8.

Animals↗

PubMed1992

Use of antigen expressed in bacteria for detection of EBV-specific thymidine kinase antibodies in sera from patients with nasopharyngeal carcinoma.

Two cDNA clones covering the N- and C-terminal portions of the EBV BXLF1 open reading frame were selected from a cDNA library derived from P3HR1 cells. The two clones were ligated, the N-terminal untranslated region truncated, and the product inserted into an E. coli expression vector, pET3CP*. The fusion protein was expressed under control of the T7 phage phi 10 gene promoter and shown to possess thymidine kinase activity. The protein was then used as an antigen to detect antibody reactivities in serum samples of nasopharyngeal carcinoma patients and healthy blood donors. Using a 1:400 dilution of serum samples in Western blot analyses, it was possible to differentiate the reactivities of serum IgA of NPC patients and healthy donors. The prevalence of positive reactivity to EBV TK in NPC was around 84%. The test was compared to others used for early diagnosis of NPC and was able to detect some patients who were negative in those tests.

Antibodies, Viral↗

PubMed1992

Cloning and expression of a cDNA encoding the Epstein-Barr virus thymidine kinase gene.

A clone of the Epstein-Barr virus (EBV) thymidine kinase (TK) gene was derived from a cDNA library of P3HR1 cells. The gene product was expressed as a fusion protein in a procaryotic system by using T7 RNA polymerase. The recombinant TK showed a molecular mass of 67 kDa and was biologically active. Antiserum raised in mice immunized with partially purified TK recognized an antigen present in EBV-superinfected Raji cells using an indirect immunofluorescence assay.

Animals↗

PubMed1991

Cloning and characterization of cDNA clones corresponding to transcripts from the BamHI G region of the Epstein-Barr virus genome and expression of BGLF2.

A cDNA library was constructed from poly(A)+ RNA isolated from the iododeoxyuridine-treated P3HR1 cell line. Five cDNA clones, which hybridized with the BamHI G fragment of Epstein-Barr virus (EBV) DNA, were subcloned and sequenced. Clones G2, G3 and G4 corresponded to the BGLF2 open reading frame (ORF) of EBV (B95-8, nucleotides 126,837 to 125,866); G3 was found to contain the entire BGLF2 ORF. The predicted Mr of the putative protein product of the EBV B95-8 BGLF2 ORF is 36K. Complete nucleotide sequencing of G3 revealed that there were two nucleotide changes from the reported sequence of the EBV B95-8 BGLF2 gene, but these did not alter the predicted amino acid sequence of the products. Clone G3 and a cDNA derived from it by N-terminal deletion were expressed in Escherichia coli, producing fusion proteins. Rabbit antisera against these proteins were shown to react with viral capsid antigen-expressing HR1 cells in an indirect immunofluorescence assay. In vitro transcription/translation products and fusion proteins expressed in E. coli were used to determine the presence of antibodies in sera from EBV-infected individuals. The results of immunoprecipitation and immunoblotting studies showed that the majority of EBV-seropositive individuals mount a serum antibody response to the BGLF2 ORF-encoded protein.

Amino Acid Sequence↗

PubMed1991

Glucosyltransferase gene polymorphism among Streptococcus mutans strains.

Genetic polymorphisms in genes coding for the glucosyltransferases were detected among Streptococcus mutans serotype c strains by Southern blot analysis with DNA probes located within the gtfB gene (H. Aoki, T. Shiroza, M. Hayakawa, S. Sato, and H. K. Kuramitsu, Infect. Immun. 53:587-594, 1986). Restriction endonucleases were used to examine genomic DNAs isolated from serotype a to h strains. The variations were readily detected among 33 strains of serotype c by EcoRI and PstI restriction enzyme digestions. Serotypes e and f, which are genetically similar to serotype c, also had comparable polymorphism; however, serotypes a, b, d, g, and h did not hybridize to the same DNA probes in parallel experiments. Further analysis of enzymatic activities for glucan synthesis and sucrose-dependent adherence revealed no significant differences among the serotype c strains. Our results suggested that genetic polymorphisms existing in S. mutans serotype c strains may reflect a complexity in genes coding for the glucosyltransferases, which are produced ubiquitously in members of the S. mutans group.

Bacterial Adhesion↗

PubMed1991

Detection of human herpesvirus 6 DNA in T and B cell lymphomas.

DNA extracted from tissues of 109 patients with lymphoproliferative disorders, including 40 T-cell lymphomas, 40 B-cell lymphomas, 8 Hodgkin's disease, 13 hyperplasia, 5 infectious mononucleosis, 2 histiocytic medullary reticulosis and 1 Kikuchi's disease, and peripheral blood lymphocytes (PBL) from 17 healthy donors were investigated for the presence of human herpesvirus 6 (HHV-6) DNA sequence by polymerase chain reaction (PCR) and Southern hybridization. The procedures for DNA amplification were composed of the first round of PCR using a pair of primers LL and LR, which were contained in the HHV-6 major capsid gene, and the second round of nested PCR using an internal pair of primers LLN and LRN. Results showed that the prevalence of HHV-6 DNA sequence in human tissues was similar in both B and T cell lymphoma patients and normal adults, and viral heterogeneity was detected in the amplified DNA sequence from local strain of Taiwan as compared to that of U1102 strain from Uganda.

DNA, Viral↗

PubMed1991

Analysis of Epstein-Barr virus DNA in nude mouse-passaged nasopharyngeal carcinoma tissues.

DNAs extracted from two series of nude mouse-passaged nasopharyngeal carcinoma (NPC) tissues, NPC-295 and -306, and P3HR1 cells were digested with restriction enzyme Bam HI, then electrophoresed on agarose gel and analysed for the Epstein-Barr virus (EBV) DNA fragments by Southern hybridization with 27 radioactive Bam HI DNA fragments and the Eco RI Dhet DNA fragment from cloned EBV DNA of B95-8 cell line. Constant hybridization patterns were revealed in these experiments and the results suggest a stable existence of EBV DNA including replicative form and episomal form in these tissues even after a long time of passage. Neither deletion in YH region nor in I region could be detected when using Bam HI Y, H and I as probes, suggesting the EBV in these tissues are "more complete" viruses. When Bam HI F was used as a probe, an extra Bam HI site was revealed in the analysis of the DNA from NPC-295 suggesting the polymorphism of DNA present in this region. Restriction maps of EBV DNAs in both nude mouse-passaged NPC tissues are proposed. The results suggest that nude mouse-passaged NPC tissue is a good model to study the structure of EBV DNA in tumor tissue.

Adult↗

PubMed1991

[Emergency obstetric hysterectomy: report of fifteen cases].

From June 1978 to June 1990, there were 41,989 deliveries, 13,396 cesarean sections and 15 emergency obstetric hysterectomies at Taipei Chang Gung Memorial Hospital. The incidence of emergency obstetric hysterectomy is 36/100,000 deliveries. The mean age of patients is 31.7 years old. The indications for hysterectomy were placental disorder (60.0%), uterine atony (26.7%), and uterine rupture (13.3%). The present series showed that 14 cases had total abdominal hysterectomy, while only one case had a subtotal hysterectomy. Total hysterectomy should generally be performed unless maternal instability mandates a more expeditious subtotal hysterectomy. The most common postoperative complication included unknown fever and urinary tract infection. For those four cases of post-cesarean hysterectomy, conservative treatment was tried but failed. All cases presented with hypotension (less than 80/60 mmHg), tachycardia (greater than 100 times/min) and with a mean hemoglobin of 9 mg% during the observation period. In these patients, blood loss, operating time and length of hospitalization was increased when compared with a group of 7 patients undergoing cesarean hysterectomy. In patients with placenta accreta, who are candidates for cesarean hysterectomy, initial hypotension, tachycardia and shock developed during cesarean section. It must be emphasized that conservative aggressive measures to control obstetric hemorrhage remain the mainstay of therapy, but emergency hysterectomy played the life-saving role with which every practitioner of obstetrics must be familiar and keep in mind.

Adult↗

PubMed1991

Human T-lymphotropic virus type I genome in adult T-cell leukemia/lymphoma.

The proviral DNA of human T-lymphotropic virus type I (HTLV-I) was examined in 8 patients with adult T-cell leukemia/lymphoma (ATL/L) by Southern blot analysis. Judging from the number of bands containing viral DNA sequences in tumor cells, one and three copies of proviral genome were detected in 7 and 1 cases, respectively. The proviral genome in 1 case contained variant PstI sites and a single defective provirus which lost 5' LTR, gag and a part of pol genes was found in 2 cases. However, the pX region was detected in both of 2 latter cases by using the polymerase chain reaction. The results indicate that not all tumor cells from ATL/L yield infectious viruses and also support that the product of pX region may contribute to the tumorigenesis of HTLV-I.

Adult↗

PubMed1990

Evolutionary conservation of target sequences for cis-acting regulation in c-myc exon 1 and its upstream region.

Transcriptional control of the c-myc proto-oncogene, an important factor in cellular growth, differentiation and in the genesis of various neoplasms, is mediated by multiple positive and negative regulators in the 5' end region of the gene. Here, we report the nucleotide sequence of the first c-myc exon and its upstream region from woodchuck, a rodent which can develop liver tumors associated with c-myc activation [Möröy et al., Nature 324 (1986) 276-279]. Alignment of these sequences with the corresponding human and murine regions shows a surprisingly high homology between woodchuck and human, and suggests the absence of species-specificity in the fundamental regulatory elements which govern c-myc expression.

Animals↗