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Biomedical subjects

H L Chen

Publications and source records attributed to H L Chen.

At least 109 records · Page 6Linked to original sources
PubMed1997

[Primary surveillance of spotted fever group antibodies on rats in the Kinmen area].

The positive rate of rickettsial antibodies of 107 rats in the Kinmen area by indirect immunofluorescent antibody (IFA) technique was 0% (0/107) in typhus fever, 38.3% (41/107) in scrub typhus and 66.4% (71/107) in spotted fever group; the positive rate (42.9%) of spotted fever group of 21 rats in Taiwan island also higher than scrub typhus (19.0). It suggests that spotted fever group patients may be present in our country but have not been discovered.

Animals↗
PubMed1997

[Effect of retinoic acids on phosphatidylcholine specific phospholipase D in 7721 human hepatocarcinoma cell line].

In order to elucidate the relation between cell signal transduction and cell differentiation, the effect of two differentiation inducers--all trans-retinoic acid (ATRA) and 13 cis-retinoic acid (13 cis-RA) on the specific activities (nmol/hr.mg protein) of phosphatidyl choline specific phospholipase D (PC-PLD) in 7721 human hepatocarcinoma cell line was studied. It was found that ATRA and 13 cis-RA increased the specific activities of membrane bound PC-PLD on 2nd and 4th day of cell culture respectively. If the PC-PLD activities were calculated as activity per bottle cells, the effect of ATRA on the 2nd day was also higher than that of 13 cis-RA, while the reverse was true on the 4th day, revealing a postponed effect of 13 cis-RA as compared with ATRA. The increase of PC-PLD was higher when the culture medium was refreshed every day than that when the medium was unrefreshed, suggesting that a factor might be existed in the fresh medium which could enhance the stimulating effect of retinoic acids on PC-PLD. The effect of ATRA on PC-PLD specific activity were higher on 2nd day than that on 4th day, owing to the accelerated proliferation of the cells on 4th day, leading to the increase of protein contents and decrease of the enzyme specific activity calculated on the base of protein contents. Whereas, the effect of 13 cis-RA on PC-PLD specific activity were higher on 4th day than that on 2nd day, because the increase of enzyme activity was higher than the increase of protein contents on 4th day. The relation between the elevation of PC-PLD and protein kinases was further studied and discovered that ATRA decreased both the specific activities of membrane bound and cytosolic protein kinase C (PKC) as well as tyrosine protein kinase (TPK) either in 2nd or 4th day of cell culture, which indicated that the increasing effect of ATRA on membrane bound PC-PLD was not resulted from the stimulating effect of PKC and TPK on PC-PLD, and its mechanism remains to be further investigated.

Antineoplastic Agents↗
PubMed1997

[Changes of PKC isoforms in induced differentiation of HL-60 cells by ATRA and PMA].

Human premyeloid leukemic cell line (HL-60) were differentiated along granulocytic or monocytic/phagocytic lineage respectively by the treatment of all-trans retinoic acid (ATRA) or phorbol ester (PMA), which was demonstrated by morphological observation, NBT reduction test, specific and nonspecific assay of esterases. By use of immuno-histochemical method, the changes of protein kinase C (PKC) isoforms alpha, beta I and beta II after induced differentiation of HL-60 cells were studied. The results showed that ATRA led to 5.0, 2.8 and 4.2 times increase of PKC alpha, beta I and beta II isoforms respectively as compared with the undifferentiated control levels and translocation of PKC from membrane to cytosol. The level of PKC alpha and beta II isoforms in HL-60 cells induced by PMA were decreased, but the expression of PKC beta I was increased. Besides, PMA led these 3 kinds of PKC to be present in cell nucleus with different intensity. These results suggest that the sustained activation of PKC alpha and beta may be required for granulocytic differentiation of HL-60 cells, while nuclear translocation of PKC may relatively be an important step for differentiation of HL-60 cells to monocytes/macrophages.

Cell Differentiation↗
PubMed1997

Nephrogenic adenoma of the ureter--a case report.

We present a case of nephrogenic adenoma, a rare benign lesion arising from the ureter. Histologically, it showed the formation of epithelial lined tubules resembling the renal tubules. Special Stain demonstrated the presence of intraluminal PAS-positive materials.

Adenoma↗
PubMed1997

Diagnosis of primary liver cancer using lectin affinity chromatography of serum alkaline phosphatase.

Serum alkaline phosphatase (sALP) can be separated into unbound liver type (L-ALP) and bound bone type (B-ALP) by means of WGA affinity chromatography. The L-ALP from the sera of normal adults and various liver diseases was found to show different chromatographic behaviours on DSA affinity column with multiple peaks of ALP activity after the L-ALP was treated with neuraminidase to remove the terminal sialic acids on the sugar chain of L-ALP. The L-ALP from normal sera contained no bound activity on DSA, whereas that from acute or chronic hepatisis, liver cirrhosis and biliary obstruction had a significant amount of bound fractions with weak and intermediate affinity. The strongly bound fraction(s) was only present in the L-ALP serum from primary liver cancer (PLC) and the positive rate of its appearance was 100% in 38 cases of PLC, including 8 alpha-fetoprotein (AFP) negative cases. The WGA chromatography can be skipped and similar results are obtained. The percentage of the strongly bound fraction in serum L-ALP was not related to the level of either sALP activity or AFP, and the appearance of the strongly bound fraction is attributed to the structural difference of sugar chains in L-ALP. Therefore, this L-ALP fraction may be assumed as a new index in the diagnosis of PLC, and the different profiles of sALP or L-ALP on DSA chromatography may be used in the differential diagnosis of benign and malignant liver diseases.

Alkaline Phosphatase↗
PubMed1997

Characterization of the allergen Der f 7 from house dust mite extracts by species-specific and crossreactive monoclonal antibodies.

BACKGROUND: The group 7 mite allergens react with IgE in 50% of sera from allergic patients. OBJECTIVE: To determine the molecular and antigenic characteristics and heterogeneity of Der f 7 in mite extracts. METHODS: Monoclonal antibodies (MoAbs) produced from mice immunized with recombinant Der f 7 were examined for crossreactivity to Der p 7 and then used for immunoblotting of 1 and 2-D gel electrophoresis. Deglycosylation was studied with N-glycosidase-F and N-terminal sequencing by Edman degradation. The epitopes of the monoclonal antibodies were compared by cross-inhibitory immunoassays. RESULTS: Immunoblotting of D. farinae extracts with all the anti Der f 7 MoAbs showed major reactivities at 31, 30 and 25 kDa. The strongest immunostaining was at 25 kDa which contrasted with Der p 7 where the 31 and 30 kDa bands were strongest. The relative strength of staining however varied between extracts. The 31 and 30 kDa components were glycosylation products of the 25 kDa form which had the N-terminal sequence predicted from cDNA analysis. Two MoAbs stained an 18 kDa band consistent with a degradation product. The 2-D gels showed that different components with pIs from 5.6-6.4. Both species-specific and Der p 7 crossreactive MoAbs were produced and a two-site ELISA assay for detecting group 7 allergen was developed with MoAbs recognizing different epitopes. CONCLUSIONS: Der f 7 has been defined by its natural N-terminal sequence and MoAbs. It apparently exists as different glycosylation and degradation products in mite extracts, the relative abundance of which differs with different preparations. A two-site ELISA to measure the allergen was developed.

Allergens↗
PubMed1997

Two novel glucose 6-phosphate dehydrogenase deficiency mutations and association of such mutations with F8C/G6PD haplotype in Chinese.

Glucose 6-phosphate dehydrogenase (G6PD) deficiency is a common genetic disease affecting 3% of the total Chinese population in Taiwan. To investigate the molecular basis of this deficiency, we analyzed blood samples from G6PD-deficient newborns using a nonradioactive polymerase chain reaction coupled with single-stranded conformation polymorphism (PCR-SSCP) analysis. We identified two novel G6PD mutations in Chinese. The first, G6PD Miaoli, involved a C-->G substitution at nucleotide (nt) 519, producing a Phe173 to Leu change in the protein. The second mutation (G6PD Keelung) involved a C-->T change at nt 1387, resulting in an Arg463 to Cys substitution. The F8C/G6PD (coagulation factor VIIIc) haplotype that spans the Xq28 region from the gene for coagulation factor VIIIc to the gene for G6PD was also investigated in Chinese using PCR and restriction enzyme digestion. Of the 16 possible haplotypes, only four were found, which suggests that these four polymorphic sites are in strong linkage disequilibrium. Analysis of the association of G6PD mutations with F8C/G6PD haplotype revealed that nt 517, 592, 835, and 1387 mutations are linked to haplotype VI+VII, whereas the nt 519 mutation is linked to haplotype III. The finding that some G6PD mutations are associated with a particular F8C/G6PD haplotype may be useful for future population studies.

Female↗
PubMed1996

A novel functional role for apolipoprotein B in male infertility in heterozygous apolipoprotein B knockout mice.

Male infertility, affecting as many as 10% of the adult population, is an extremely prevalent disorder. In most cases, the cause of the condition is unknown, and genetic factors that might affect male fertility, other than some sequences on the Y chromosome, have not been identified. We report here that male mice heterozygous for a targeted mutation of the apolipoprotein B (apo B) gene exhibit severely compromised fertility. Sperm from these mice failed to fertilize eggs both in vivo and in vitro. However, these sperm were able to fertilize eggs once the zona pellucida was removed but displayed persistent abnormal binding to the egg after fertilization. In vitro fertilization-related and other experiments revealed reduced sperm motility, survival time, and sperm count also contributed to the infertility phenotype. Recognition of the infertility phenotype led to the identification of apo B mRNA in the testes and epididymides of normal mice, and these transcripts were substantially reduced in the affected animals. Moreover, when the genomic sequence encoding human apo B was introduced into these animals, normal fertility was restored. These findings suggest that this genetic locus may have an important impact on male fertility and identify a previously unrecognized function for apo B.

Animals↗
PubMed1996

Seroepidemiology of hepatitis B virus infection in children: Ten years of mass vaccination in Taiwan.

OBJECTIVE: To study the seroepidemiology of hepatitis B virus (HBV) infection in children 10 years after a mass hepatitis B vaccination program was begun in Taiwan. DESIGN: Cross-sectional seroprevalence survey. SETTING: Cheng-Chung/Chung-Cheng District, Taipei, Taiwan, 1994. SUBJECTS AND METHODS: Serum samples from 1515 healthy children younger than 12 years were tested for HBV markers. The results were compared with a baseline seroepidemiologic study conducted just before the vaccination program was launched in 1984 and with a subsequent study in 1989 in the same area. MAIN RESULTS: Eighty-seven percent of the children had received at least 3 doses of HBV vaccine. The overall prevalence rate of hepatitis B surface antigenemia decreased from 9.8% in 1984 to 1.3% in 1994. A statistically significant decrease was observed in every age group from 1 to 10 years. The overall prevalence rate of hepatitis B core antibody was 26% in 1984, 15% in 1989, and 4.0% in 1994. This suggests that the risk of horizontal HBV infection has decreased over time, not only because of the protective effect of the vaccine but also because the infection source has diminished. A high prevalence rate of hepatitis B surface antibody (79%) was noted in 1994 as anticipated. CONCLUSIONS: The Taiwanese mass vaccination program has protected most children younger than 10 years from becoming carriers, reducing both perinatal and horizontal HBV transmission. Mass HBV vaccination has proved to be a successful method to control HBV infection in this hyperendemic area.

Child↗
PubMed1996

Structural and functional analysis of beta2 microglobulin abnormalities in human lung and breast cancer.

The escape of tumor cells from immune recognition is a central problem in tumor immunology. Here, we examined the functional role of somatic beta 2-microglobulin (beta2m) gene mutations in human lung and breast cancers. Using single-strand conformational polymorphism (SSCP) analysis and DNA sequencing, we found mutations in the beta2m gene in 2 of 110 tested lung, colon and breast tumors and tumor cell lines. No mutations were identified in 63 breast cancer tumors, in B-lymphoblastoid cell lines or normal tissues from these or other patients. In these cell lines, beta2m protein was undetectable by Western blot analysis and there was no MHC class I on their cell surface even after treatment with interferon-gamma. Transfection of these tumor cell lines with the beta2m gene, but not addition of purified beta2m protein restored MHC expression without addition of exogenous pepticles, indicating that endogenous beta2m expression is necessary for proper intracellular MHC assembly and stabilization by endogeneous pepticles. Mutation in beta2m caused cell line H2009 to be resistant to specific lysis by influenza virus-specific CTL from HLA matched donors, and transfection of the beta2m gene restored this killing. A small cell lung cancer cell line with low class I expression and with a normal beta2m genomic sequence nonetheless also demonstrated increased class I expression after transfection of the beta2m expression vector alone, indicating that the availability of beta2m may be rate limiting for MHC assembly in this line. Our results indicate that somatic mutations or selective loss of expression of the beta2m gene in human lung cancer is rare, but can cause defective MHC class I expression and function allowing these cells to escape recognition by cytotoxic T cells.

Breast Neoplasms↗
PubMed1996

Calorimetric characterization of the formation of acrylic type bone cements.

The formation of acrylic bone cements upon heating was investigated by differential scanning calorimetry (DSC). The effects of the contents of initiators, accelerator, biocompatibilizer, and crosslinking agents on the rate and the heat of polymerization during DSC heating were studied. The rate and the heat of polymerization (delta H) were characterized by the peak temperature and the area of the DSC exotherm, respectively. It was found that both the rate and heat of polymerization decreased with increasing heating rate. The delta H was increased considerably with increasing benzoyl peroxide (BPO) initiator concentration from 1 to 10% (w/v), whereas the rate of polymerization was reduced significantly. An increase in azobisisobutyronitrile (AIBN) initiator concentration also induced an increase in delta H, but the rate of reaction was not affected considerably. The addition of accelerator promoted the rate of reaction but resulted in a drop in delta H. The rate of polymerization for the system containing BPO initiator was increased quite significantly with the addition of hydroxyethyl methacrylate (HEMA) biocompatibilizer, while the delta H was slightly increased. For the system using AIBN as the initiator, the rate of polymerization was decreased slightly and the delta H dropped significantly with the addition of HEMA. The effect of ethylene glycol dimethacrylate (EGDMA) crosslinking agent was also examined. Polymerization became more rapid with the addition of EGDMA in the bone cement using BPO initiator, while it remained approximately constant for the system using AIBN as the initiator. No systematic change in delta H was observed with the addition of EGDMA in both systems. This study demonstrated that DSC is a potential tool to measure the amount of heat released and also the rate of polymerization for bone cements.

Acrylates↗
PubMed1996

An IgG-transporting Fc receptor expressed in the syncytiotrophoblast of human placenta.

During normal human pregnancy, maternal IgG crosses the placenta and provides passive immunity for the fetus. In so doing, IgG passes through two cellular barriers: the syncytiotrophoblast and the fetal capillary endothelium. The Fc region of IgG is required for its transport across the placenta, but the Fc receptors responsible have not been identified definitively. We recently reported the isolation from a placental cDNA library of clones encoding the alpha chain of a human homologue of the major histocompatibility complex class I-related Fc receptor, the neonatal Fc receptor (FcRn). In mice, FcRn is essential for the transport of maternal IgG to the fetus and the neonate. We report here the localization of human FcRn mRNA within the placenta by in situ hybridization, and of human FcRn protein by immunohistochemistry. Both methods show that human FcRn is expressed in syncytiotrophoblast, and is, thus, appropriately located to transport maternal IgG across the first barrier. We confirm previous findings that specific binding of IgG to placental membranes is greater at pH 6.0 than pH 7.5. This corresponds with the pH dependence of IgG binding to FcRn and is consistent with the presence of FcRn in syncytiotrophoblast. We propose a transport model in which maternal IgG binds FcRn at low pH in endosomes within the syncytiotrophoblast. FcRn is not expressed in fetal capillary endothelia, and the mechanism of IgG transport across the second barrier remains unknown.

Amino Acid Sequence↗
PubMed1996

Analysis and comparison of the mouse and human immunoglobulin heavy chain JH-Cmu-Cdelta locus.

We report here 23,686 bases of contiguous DNA sequences from the mouse germline immunoglobulin heavy chain (H) constant (C) mu delta region. The sequence spans the joining (JH) regions, the mu constant region (C mu), the delta constant region (C delta) coding regions, a domain relic, the mu switch region (S mu), seven blocks of simple sequence repeats, a large unique sequence inverted repeat, a large unique sequence forward repeat, and all of the intervening material. A comparison of this 23.7-kb region with the corresponding human C mu/C delta region reveals clear homology in the coding and introns of C mu but not in the 5' flanking J gene segments nor in the intergenic and C delta regions. This mixed pattern of similarity between the human and the mouse sequences contrasts with high levels of similarity found in the T-cell receptor C alpha/C delta region and alpha and beta myosin genes and the very low levels found in the gamma-crystallin, XRCC1, and beta-globin gene clusters. The human and mouse comparison further suggests the incorporation of novel sequences into expressed genes of IgD.

Animals↗
PubMed1996

Late signals are required for the stimulation of DNA synthesis in rat mammary fibroblasts by growth factors.

Maximal stimulation of DNA synthesis in quiescent rat mammary (Rama) 27 fibroblasts is elicited by epidermal growth factor (EGF) or basic fibroblast growth factor (bFGF) 18 h after the initial addition of the growth factors-the 'lag' period. At maximally-stimulating concentrations, EGF and bFGF are interchangeable 9 h after their initial addition. When the initial concentration of growth factor is below that required to elicit a maximal response, it is possible to increase the level of DNA synthesis by increasing the concentration of growth factor 9 h after its initial addition. When the initial concentration of growth factor is high, substitution by a lower concentration of growth factor after 9 h allows a greater proportion of cells to synthesize DNA than would be expected from a continuous low dose of growth factor. Similar results are obtained when both the growth factor and its concentration are changed 9 h after the initial addition of growth factor. However, when EGF at a low concentration is substituted for a high concentration of EGF or bFGF the resulting increase in the levels of DNA synthesis is greater when EGF rather than bFGF is added for a second time. The half-life of the growth-stimulatory signals delivered by EGF and by bFGF 9 h after their initial addition is 1-2 h. These results suggest that to stimulate DNA synthesis: (i) EGF or bFGF must deliver a signal(s) continuously; (ii) the initial signals produced by EGF and bFGF are equivalent; (iii) the signals produced between 9-18 h by EGF may be different to those produced by bFGF.

Animals↗
PubMed1996

Chromosome 1p aberrations are frequent in human primary hepatocellular carcinoma.

To investigate the chromosomal changes in hepatocellular carcinoma in humans, metaphase chromosomes obtained directly from short-term culture of resected human primary hepatocellular carcinoma tissues were studied in seven patients. Among the five successfully karyotyped cases, cytogenetic analysis showed highly different and complex chromosomal changes in the cancer tissue in four patients. Structural aberrations of chromosome 1 and deletion of 1p with breakpoints at p22 or p32 were the most common abnormalities, and were found in the four successfully analyzed cases. Loss of 1p may be important in hepatocarcinogenesis, and warrants further characterization by molecular genetic analysis.

Adult↗
PubMed1996

A functionally defective allele of TAP1 results in loss of MHC class I antigen presentation in a human lung cancer.

Tumours express a variety of novel epitopes which represent potential immune targets, and thus clinically evident tumours are thought to have effectively avoided immune recognition and elimination. Transporters associated with antigen presentation (TAP) are thought to be responsible for conveying intracellular peptides into the endoplasmic reticulum for complex formation with class I MHC and subsequent recognition by cytotoxic T lymphocytes. In this study, we evaluated 79 human solid tumours and cell lines for genetic abnormalities in TAP1 that might have led to an acquired loss of antigen presenting ability. A novel sequence (R659Q) was discovered near the ATP binding site in a human small cell lung cancer (SCLC) cell line, H1436. This cell line is heterozygous for this allele, but only the R659Q allele is transcribed into RNA. Even though the R659Q protein is expressed, these cells act as if they were TAP deficient by peptide binding and antigen presentation studies, which are restored after transfection of a functional TAP1 allele. This is the first evidence for a naturally occuring protein structural defect resulting in defective peptide transport in a human solid tumour.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1996

Production of vascular endothelial growth factor by human tumors inhibits the functional maturation of dendritic cells.

Inadequate presentation of tumor antigens by host professional antigen-presenting cells (APCs), including dendritic cells (DCs), is one potential mechanism for the escape of tumors from the host immune system. Here, we show that human cancer cell lines release a soluble factor or factors that dramatically affect DC maturation from precursors without affecting the function of relatively mature DCs. One factor responsible for these effects was identified as vascular endothelial growth factor (VEGF). Thus, VEGF may play a broader role in the pathogenesis of cancer than was previously thought, and therapeutic blockade of VEGF action may improve prospects for immunotherapy as well as inhibit tumor neovasculature.

Antigen Presentation↗