Biomedical subjects

H T Chen

Publications and source records attributed to H T Chen.

At least 37 records · Page 2Linked to original sources

PubMed1998

Growth hormone receptor and regulation of gene expression in fetal lymphoid cells.

The role of growth hormone (GH) in modulating the adult immune response is receiving increased attention; however, its role in the development of immune competence in the fetus has not been defined. In order to begin to address the role of GH in the ontogeny of the immune response. cells from bovine fetal spleen and thymus were examined for GH receptor and responsiveness to GH. Northern analysis and ligand binding studies showed that growth hormone receptor (GHR) was readily detected in early- and mid-gestational fetal thymocytes, but it was less readily detected in thymocytes from older fetuses. In contrast, GHR was easily detected in splenocytes at all fetal ages. Thymocytes and splenocytes from mid-gestational fetuses expressed low levels of cell surface GHR by flow cytofluorometric analysis, and CD4+ and CD8 (single positive) thymocyte subsets were positive. Northern analyses were employed to determine the effects of in vitro GH treatment on expression of several proto-oncogenes, cytokines, and GHR in thymocytes from fetuses at approximately mid-gestation. GH treatment for 30 min down-regulated c-jun and c-fos mRNA approximately 2- and 2.8-fold, respectively. After 6 h treatment, GH increased transcript levels for interleukin (IL)-1alpha, IL-1beta, IL-6, and GM-CSF about 2.5-, 2.2-, 3-, and 2-fold, respectively. GH also down-regulated the expression of its own receptor about 3.2-fold after 8 h of incubation. The presence of GHR in fetal lymphoid cells and its temporal and spatial regulation suggest a potential role for GH in the development and or function of the fetal bovine immune system. Although the mechanism(s) is unclear, our results suggest that GH is intimately involved in lymphocyte function and expression of certain cytokines during a critical period of fetal immune development.

Animals↗

PubMed1997

Molecular characterization of the bovine type II IL-1 receptor.

The purpose of this study was to clone and characterize the bovine IL-1rII. Using a human IL-1rII probe, three putative exons of the bovine IL-1rII were identified from a bovine genomic library. The full length cDNA for bovine IL-1rII was cloned from a bovine endometrial cDNA library using oligonucleotides designed from those exons. The nucleotide sequence of the bovine IL-1rII cDNA has 79, 69 and 69% identity with those of human, mouse, and rat IL-1rII, respectively. The bovine IL-1rII cDNA contains an open reading frame encoding 400 amino acids with a predicted amino acid sequence of 71, 58, and 59% identity with those of human, mouse and rat IL-1rII, respectively. Transient transfection of COS cells with the cloned bovine IL-1rII cDNA resulted in specific binding of bovine IL-1 beta, with a Kd of 1.82 x 10(-10) M. At least two transcripts of bovine IL-1rII were identified from bovine peripheral blood mononuclear cells (PBMCs), liver, and fetal thymocytes by Northern blot analysis. The relative proportion of these transcripts varied with cell type. In addition, bovine IL-1rII transcripts were detected by RT-PCR in stimulated and unstimulated neutrophils, fibroblasts, and splenocytes. The results of this study provide the first characterization of bovine IL-1rII at a molecular level.

Amino Acid Sequence↗

PubMed1997

Free tissue transfer for leg reconstruction in children.

Twenty-five free tissue transfers for reconstruction after lower limb trauma or release of scar contractures were performed in 23 children over an 8-year period. The ages ranged from 2 to 14 years (mean 7.1 years). Fourteen (61%) patients had major associated injuries. The reconstruction sites were all in the distal leg. The overall vascular success rate was 96%. Three patients underwent re-explorations because of vascular insufficiency. One flap failed and two were salvaged. The causes of vascular complications were considered to be inadequate debridement of an avulsed vessel, venous kinking and inadequate protection of the vascular pedicle. The three cases with vascular insufficiency were children with associated injuries on the ipsilateral leg proximally. Postoperative complications were not uncommon (28%). Six of the seven complications occurred in patients with major associated injuries. The mean follow-up was 57 months. Long-term problems such as a limping gait, frequent flap ulcers, toe contracture or a bulky flap were common. Our results suggest that free tissue transfer is reliable and safe for the reconstruction of major leg injuries in children. Associated injuries proximal to the reconstructed defects on the ipsilateral leg predispose to more vascular complications. Since postoperative complications and long-term morbidity are common in children, special precautions should be taken with the preoperative assessment, planning, execution of surgery and postoperative care to achieve better results.

Accidents, Traffic↗

PubMed1997

Growth hormone and Pit-1 expression in bovine fetal lymphoid cells.

Bovine fetal lymphoid cells were examined for growth hormone (GH) and the transcription factor Pit-1/GHF-1 mRNA. GH and Pit-1/GHF-1 transcripts were detected in thymocytes and splenocytes from fetuses at 60, 90, 120, and 270 d of gestation using reverse transcription-polymerase chain reaction (RT-PCR). Northern analysis indicated that the lymphoid GH mRNA was approximately 350 nucleotides larger than in the pituitary. RT-PCR analysis demonstrated that the coding regions as well as 3' untranslated region of the lymphocyte GH and pituitary transcripts were the same. Analysis of the 5'-untranslated region of the lymphocyte GH mRNA showed that transcription began upstream from the start site in the pituitary gland, suggesting differences in regulation in these tissues. Fetal thymocytes and splenocytes expressed Pit-1/GHF-1 mRNA; however, they contained only the 2.5-kb transcript. The GH and Pit-1/GHF-1 mRNA in fetal lymphoid cells supports the hypothesis that lymphocyte-derived GH may function as an autocrine and/or paracrine factor during the development and maturation of the bovine fetal immune system.

Animals↗

PubMed1996

Effect of chloral hydrate on in vivo KCl-induced striatal dopamine release in the rat.

The release of striatal dopamine (DA) and its metabolites in response to locally-induced K+ depolarization was investigated in vivo in chloral hydrate-anesthetized and freely moving rats. KCl at concentrations of 30, 50, and 100 mM induced significant dose-dependent increases in extracellular DA overflow in both chloral hydrate-anesthetized and freely moving rats (P < 0.05). Extracellular levels of dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindoleacetic acid (5-HIAA) were decreased. The DA overflow in response to 30 mM KCl stimulation in anesthetized rats was significantly greater than that in freely moving rats (P < 0.05). In addition, chloral hydrate anesthesia resulted in a significant decrease in extracellular levels of DOPAC and significant increases in extracellular levels of HVA and 5-HIAA in comparison with freely moving rats (P < 0.05). Furthermore, the basal level of extracellular HVA in chloral hydrate-anesthetized rats was significantly higher than that in freely moving rats. These results suggest that chloral hydrate anesthesia could have significant effects on the pharmacological response of the striatal dopaminergic neurons.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1996

Rabbit DQ52 and DH gene expression in early B-cell development.

Rabbits predominantly rearrange the most 3'VH gene (VH1); thus combinatorial diversity is very limited. In man and mouse, the most 3'DH gene, DQ52, is preferentially rearranged early in B-cell development. To test whether this preference for rearranging a DH gene segment based on 3' end proximity exists in rabbit, we cloned and sequenced the rabbit DQ52 gene. The 11 base pair coding region sequence is identical to a published mouse DQ52, and 81.8% similar to the human sequence. It is localized approximately 805 bp upstream of the JH1 gene. However, the 3' recombination signal sequence has an atypical nonamer. We prepared mRNA from 15- to 28-day fetal rabbits and amplified expressed VDJ sequences of mu mRNA by RT-PCR. The PCR products with VDJ rearrangements were cloned and sequenced. As expected, 44 of 45 VDJ sequences reflected use of the 3' VH1a2 gene, but the DQ52 gene was utilized very infrequently, if at all. We found only one VDJ sequence from 28-day fetal liver B-cells with 8 bp that matched the germline DQ52 sequence. Instead of expressing DQ52, another DH gene, Df was frequently expressed. We cloned the genomic Df gene and localized it about 32 kb upstream of the JH region. Thus, in contrast to man and mouse, rabbits preferentially express a DH gene located in the middle of the DH region early in B cell ontogeny. This may correlate with more frequent initial rearrangement of VH to DH in rabbit B cells.

Amino Acid Sequence↗

PubMed1996

The salvage of a degloved hand skin flap by arteriovenous shunting.

Nine cases of hand degloving injuries were treated successfully with arteriovenous shunting technique. Of these nine cases, four were degloved from the wrist level and one from the forearm, three were degloved at the palm and one at the dorsum of the hand. All injuries resulted in distally based skin flaps, which were either superficial to the palmar fascia or within the subcutaneous layer. Lack of active bleeding from the periphery of the avulsed flaps substantiated circulatory compromise before revascularization. Survival of the avulsed flaps was achieved by directing the proximal arterial flow into the venous channel within the avulsed skin flaps. The post-operative care and rehabilitation were straightforward, and functional results were satisfactory.

Adolescent↗

PubMed1996

Effect of ionizing radiation on in vivo striatal release of dopamine in the rat.

The time-course effect of ionizing radiation on the levels of basal and KCl-stimulated striatal release of dopamine (DA) was examined in vivo using microdialysis techniques. The basal level of extracellular DA in sham-irradiated controls was 0.172 +/- 0.042 pmol/sample (n = 9), and it increased 7.1-fold after the stimulation by 30 mM KCl (20microliters). However, the release of dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), two metabolites of DA, was reduced significantly by 30 mM KCl (P < 0.05). In the presence of 10 microM forskolin, an activator of adenylate cyclase, a second stimulation by 30 mM KCl increased the release of DA 6.9-fold. Radiation exposure, at a dose of 10 Gy at 10 Gy/min, had no significant effect on the levels of either basal or KCl-stimulated release of DA or on the release of DOPAC and HVA. Striatal DA release increased in response to two consecutive challenges of KCl. However, the release of DA in response to the second challenge of KCl was significantly smaller than that after the first challenge (543 +/- 110% compared to 794 +/- 164%, P < 0.05; Student's paired t test). Pretreatment with 10 muM forskolin, which by itself had no significant effect on the level of basal release of DA, prevented the decreased response of DA to the second challenge of KCl. Our results suggest that radiation exposure at the dose we used has no significant effect on the level of the basal release of DA or the release of DA stimulated by 30mM KCl in the rat striatum, and that a reduced release of DA in response to repeated KCl stimulation might involve the cAMP effector system.

3,4-Dihydroxyphenylacetic Acid↗

PubMed1995

Characterization of a rabbit germ-line VH gene that is a candidate donor for VH gene conversion in mutant Alicia rabbits.

Normal rabbits preferentially rearrange the 3'-most VH gene, VH1, to encode Igs with VHa allotypes, which constitute the majority of rabbit serum Igs. A gene conversion-like mechanism is employed to diversify the primary Ab repertoire. In mutant Alicia rabbits that derived from a rabbit with VHa2 allotype, the VH1 gene was deleted. Our previous studies showed that the first functional gene (VH4) or VH4-like genes were rearranged in 2- to 8-wk-old homozygous Alicia. The VH1a2-like sequences that were found in splenic mRNA from 6-wk and older Alicia rabbits still had some residues that were typical of VH4. The appearances of sequences resembling that of VH1a2 may have been caused by gene conversions that altered the sequences of the rearranged VH or there may have been rearrangement of upstream VH1a2-like genes later in development. To investigate this further, we constructed a cosmid library and isolated a VH1a2-like gene, VH12-1-6, with a sequence almost identical to VH1a2. This gene had a deleted base in the heptamer of its recombination signal sequence. However, even if this defect diminished or eliminated its ability to rearrange, the a2-like gene could have acted as a donor for gene-conversion-like alteration of rearranged VH genes. Sequence comparisons suggested that this gene or a gene like it could have acted as a donor for gene conversion in mutant Alicia and in normal rabbits.

Amino Acid Sequence↗

PubMed1995

Human and mouse monoclonal antibodies to blood group A substance, which are nearly identical immunochemically, use radically different primary sequences.

A human monoclonal antibody (HuA) specific for blood group A substance with two fucose groups was found to be immunochemically almost identical with that of a previously characterized mouse monoclonal anti-A, AC-1001. The VH and VL chain cDNAs of HuA were sequenced and compared with those of AC-1001. The human and mouse antibodies used VH and Vk genes that came from different families and shared minimal nucleotide and amino acid sequence identity. Thus, two antibodies from two different species can use evolutionarily unrelated sequences to bind the same carbohydrate epitope. The cloned HuA VH and VL genes were then transfected into a mouse myeloma cell line and re-expressed, together, and each separately with an irrelevant VH or VL. Only the original HuA VH and Vk had anti-A activity, demonstrating that both the heavy and light chains contributed to specificity.

ABO Blood-Group System↗

PubMed1994

Distal phalanx replantation.

Microsurgical replantation has been attempted in the distal phalanx of the digits. Twenty-three complete, single digit amputations are reviewed. The patients' ages ranged from 9 months to 48 years. Viability was 78%. Sensory recovery averaged 7 mm static two-point discrimination. Range of motion in the distal interphalangeal joint of the finger and the interphalangeal joint of the thumb averaged 64 and 51 degrees, respectively. Eight of the 17 successfully replanted digits had mild to moderate atrophy. Eighty-two percent of patients were able to use the replanted digit successfully in their daily work. We feel that distal digital replantation is rewarding. In successful cases, it always preserves acceptable function and good appearance at a price of an average of 2.3 months off work.

Adolescent↗

PubMed1993

VH gene expression and regulation in the mutant Alicia rabbit. Rescue of VHa2 allotype expression.

Rabbits of the Alicia strain, derived from rabbits expressing the VHa2 allotype, have a mutation in the H chain locus that has a cis effect upon the expression of VHa2 and VHa- genes. A small deletion at the most J-proximal (3') end of the VH locus leads to low expression of all the genes on the entire chromosome in heterozygous ali mutants and altered relative expression of VH genes in homozygotes. To study VH gene expression and regulation, we used the polymerase chain reaction to amplify the VH genes expressed in spleens of young and adult wild-type and mutant Alicia rabbits. The cDNA from reverse transcription of splenic mRNA was amplified and polymerase chain reaction libraries were constructed and screened with oligonucleotides from framework regions 1 and 3, as well as JH. Thirty-three VH-positive clones were sequenced and analyzed. We found that in mutant Alicia rabbits, products of the first functional VH gene (VH4a2), (or VH4a2-like genes) were expressed in 2- to 8-wk-olds. Expression of both the VHx and VHy types of VHa- genes was also elevated but the relative proportions of VHx and VHy, especially VHx, decreased whereas the relative levels of expression of VH4a2 or VH4a2-like genes increased with age. Our results suggest that the appearance of sequences resembling that of the VH1a2, which is deleted in the mutant ali rabbits, could be caused by alterations of the sequences of the rearranged VH4a2 genes by gene conversions and/or rearrangement of upstream VH1a2-like genes later in development.

Amino Acid Sequence↗

PubMed1993

Evaluation issues in the Drake Chemical Workers Notification and Health Registry Study.

The Drake Chemical Workers' Health Registry combined notification of workers about bladder cancer risk with access to a free program for screening and diagnosis. Evaluation of the project has given rise to several findings and new research questions. Findings in this article illustrate the following evaluation issues: 1) studying the combination of strategies that are most effective and cost effective to notify workers of their disease risks, 2) determining the realistic yield from strategies to gain participation in health screening and other protective services for notified workers, 3) identifying the notification strategies that were most effective for different kinds of participants, 4) using process evaluation to identify key activities for ensuring continued participation of cohort members in screening, and 5) examining the extent to which participants are willing to quit smoking to protect their health.

2-Naphthylamine↗

PubMed1993

The effect and mechanism of the prokinetic action of cisapride on gastrointestinal smooth muscle.

Cisapride is a new prokinetic agent which can facilitate or restore motility throughout the entire gastrointestinal tract. Although facilitation of acetylcholine release has been suggested, the mechanism of action of cisapride is not clear. To investigate the effect and mechanism of action of cisapride, we used isolated muscle strips (with mucosa and submucosa removed) of guinea pig antrum, ileum and colon to study: (1) the dose response to cisapride, (2) the effect of antagonists (atropine and tetrodotoxin) on the stimulatory effect of cisapride. Besides these studies, we also used 3H-acetylcholine release method to investigate the acetylcholine release effect of cisapride. Cisapride elicited a dose-related enhancement of baseline activity (motility index) on the antrum and contraction on the ileum and colon at the dose of 4, 40 and 400 nM. At higher doses (4 microM) cisapride caused inhibition. This bell-shaped dose response curve suggested that cisapride might be autoinhibitory or that the receptors of cisapride might consist of high affinity stimulatory and low affinity inhibitory sites. The stimulatory responses elicited by cisapride (400 nM) were not significantly inhibited by atropine and tetrodotoxin in the antrum, ileum and colon. This suggested that cisapride might act directly on the smooth muscle. Cisapride (400 nM) evoked a rather small increase of 3H-acetylcholine release on the antrum, ileum and colon. Because the percentage of increase was small and we had demonstrated that the stimulatory effects of cisapride were not blocked by atropine and tetrodotoxin, the acetylcholine release effect of cisapride was considered unimportant.

Acetylcholine↗

PubMed1993

Microsurgical reconstruction of distal digits following mutilating hand injuries: results in 121 patients.

From 1982-1989, 152 mutilated distal digits were reconstructed with microsurgical foot tissue transfers in 121 patients. These various foot tissues included wrap-around flaps or pulp from the great toe, second toe, or third toe, as well as partial toe, nail, web space skin, and other portions of the foot. The reconstructions were carried out primarily and secondarily in 78 and 74 patients respectively. The immediate success rate was 98%. Most of the patients were satisfied with both the cosmetic appearance and the functional result. This relatively minor microsurgical foot tissue transfer proved to be an ideal reconstructive option for distal digital defects.

Adolescent↗

PubMed1993

Antitumor agents, 145. Cytotoxic asprellic acids A and C and asprellic acid B. new p-coumaroyl triterpenes, from Ilex asprella.

Three new p-coumaroyl triterpenes, asprellic acids A [1], B [2], and C [3], were isolated from the EtOH extract of dried leaves of Ilex asprella. The structures for 1-3 have been established as 3,27-di-O-trans-p-coumaroyl [1], 3-O-trans-p-coumaroyl-27-O-cis-p-coumaroyl- [2], and 3-O-cis-p-coumaroyl-27-O-trans-p-coumaroyl- [3] 3 beta,27-dihydroxyolean-12-en-28-oic acid, respectively, by spectral and chemical examinations. Asprellic acid A [1] exhibited potent cytotoxicity against the RPMI-7951 cell line with an ED50 value of 0.62 micrograms/ml, while the cytotoxicity of asprellic acid C [3] against RPMI-7951 was marginal (ED50 5.5 micrograms/ml). Compounds 1 and 3 also showed cytotoxicity against KB cells with ED50 values of 3.75 and 2.86 micrograms/ml, respectively. Asprellic acid B [2] did not show cytotoxicity (> 10 micrograms/ml) against KB or RPMI-7951 cell lines.

Adenocarcinoma↗

PubMed1993

A point mutation in the second zinc finger of the DNA-binding domain of the androgen receptor gene causes complete androgen insensitivity in two siblings with receptor-positive androgen resistance.

We have analyzed the nucleotide sequence of complementary and genomic DNAs of the human androgen receptor (AR) gene in two siblings (patients 9006 and 9030) with receptor-positive complete androgen insensitivity (Rec(+)-CAI). Northern analysis indicated that mRNA of the AR was normal in size. However, its expression was relatively reduced in both patients. Consistent with the normal androgen-binding capacity (496 and 552 fmol/mg DNA for patients 9006 and 9030, respectively) but decreased DNA-binding ability (168 fmol/mg DNA) measured in genital skin fibroblasts, no mutation was found in both N-terminal and ligand-binding domains of the AR. However, a single base substitution (G-->A) was found in the second zinc finger of the DNA-binding domain at nucleotide 2372 of the AR cDNA in both cases. This resulted in the replacement of a highly conserved arginine residue (amino acid 614) by a histidine. When the mutated receptor plasmid was cotransfected into PC-3 cells together with the reporter chloramphenicol acetyltransferase gene, chloramphenicol acetyltransferase activity was not induced by 5 alpha-dihydrotestosterone treatment, confirming that the mutation renders the AR nonfunctional and can, therefore, be held responsible for the clinical features in these patients. These results highlight the importance of Arginine-614 in the second zinc finger of the DNA-binding domain of the AR in the protein-DNA interaction.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

PubMed1992

Quantitative autoradiography of 3H-paroxetine binding sites in rat brain.

The use of 3H-paroxetine as a ligand for quantitative autoradiography of serotonin (5-HT) transport sites was optimized, and the distribution of 3H-paroxetine binding sites in rat brain was studied. Under the conditions described, 3H-paroxetine binding in forebrain sections was of high affinity and saturable, with a Kd of 0.18 +/- 0.02 nM (mean +/- SEM) and Bmax of 268 +/- 12 fmol/mg of protein (n = 3). Nonspecific binding was 10.7% +/- 1.0 of the total binding (n = 8). The distribution of 3H-paroxetine binding sites closely matched the regional distribution of 5-HT nerve terminals and cell bodies. The highest concentrations of 3H-paroxetine binding sites were found in the dorsal raphe nucleus (563 +/- 55 fmol/mg tissue, n = 4), and high densities of binding were also found in the locus coeruleus, medial forebrain bundle, substantia nigra, several limbic structures (amygdala, hippocampus, hypothalamus, olfactory tubercle, septum, and thalamus), and components of the visual relay system (superior colliculus and the lateral geniculate body). Although lesioning of 5-HT neurons with p-chloroamphetamine (PCA) drastically eliminated 3H-paroxetine binding in most regions of the rat brain, significant binding remained in the raphe nuclei and medial forebrain bundle suggesting that 3H-paroxetine binding in these regions was to presynaptic sites on cell bodies or axons relatively resistant to PCA.

Animals↗