Search PubMed⌕ Search

Biomedical subjects

J Han

Publications and source records attributed to J Han.

At least 361 records · Page 20Linked to original sources
PubMed1994

Instability of frog virus 3 mRNA in productively infected cells.

Cloned DNA restriction fragments encoding representative frog virus 3 messages were used as probes to assess the stability of viral transcripts in infected fathead minnow cells. Analysis of Northern blot hybridization profiles confirmed earlier findings and revealed that in infected cells the steady-state level of representative frog virus 3 (FV3) messages increased throughout the replication cycle. However, when actinomycin D was added at 4 hr after infection to block the synthesis of new transcripts, viral messages were observed to turn over rapidly, with half-lives of approximately 2 hr. These results indicate that viral transcripts were not preferentially stabilized in FV3-infected cells and suggest that the high steady-state level of viral messages present at late times after infection was due to viral transcription outpacing message degradation. Moreover, the instability of viral messages challenges the suggestion that the terminal dyad symmetry (hairpin structure) observed in all frog virus 3 messages sequenced to date plays a role in transcript stability.

Animals↗
PubMed1994

A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells.

Mammalian cells respond to endotoxic lipopolysaccharide (LPS) by activation of protein kinase cascades that lead to new gene expression. A protein kinase, p38, that was tyrosine phosphorylated in response to LPS, was cloned. The p38 enzyme and the product of the Saccharomyces cerevisiae HOG1 gene, which are both members of the mitogen-activated protein (MAP) kinase family, have sequences at and adjacent to critical phosphorylation sites that distinguish these proteins from most other MAP kinase family members. Both HOG1 and p38 are tyrosine phosphorylated after extracellular changes in osmolarity. These findings link a signaling pathway in mammalian cells with a pathway in yeast that is responsive to physiological stress.

Amino Acid Sequence↗
PubMed1994

Over-representation of the disease associated (CAG) and (CGG) repeats in the human genome.

Expansion of trimer repeats has recently been described as a new type of human mutation. Of the 64 possible trimer compositions, only the CGG and CAG repeats have been implicated in genetic diseases. This study intends to address two questions: (1) What makes the CGG and CAG repeats unique? (2) Could other trimer repeats be involved in this type of mutation? By computer analysis of trimer and hexamer frequency distributions in approximately 10 Mb of human DNA, twenty trimer motifs (ten complementary pairs) have been identified that are the most likely to be expanded. The frequency distribution study also indicated that the expanded trimer motif in Fragile-X syndrome is GGC instead of CGG. DNA linguistics studies revealed that the GGC/GCC and CAG/CTG repeats were over-represented in the human genome. Further analysis of base composition suggested that the CCA/TGG repeats may be involved in the trimer expansion mutation since they possessed many similar characteristics to GGC/GCC and CAG/CTG. The computer aided sequence analysis studies reported here may help to understand the molecular mechanisms of trimer repeat expansion.

Base Sequence↗
PubMed1994

Lipopolysaccharide (LPS) binding protein, truncated at Ile-197, binds LPS but does not transfer LPS to CD14.

Lipopolysaccharide (LPS) binding protein (LBP), a 58-60 kDa glycoprotein, binds to the lipid A region of LPS. The resulting LPS-LBP complex is recognized by both the membrane-bound (mCD14) and soluble forms of CD14 (sCD14), thereby enhancing the ability of LPS to activate myeloid, endothelial, and epithelial cells. To begin to characterize the structure-function relationships within LBP, we have created and expressed a truncated form of human LBP (herein called NH-LBP) comprising amino acid residues 1-197 of the parent molecule. Experiments were done to characterize the ability of NH-LBP to bind LPS and to promote LPS binding to CD14. We found that NH-LBP efficiently binds LPS but does not transfer the LPS to either mCD14 or sCD14. Additionally, NH-LBP inhibited LPS binding to LBP, inhibited the LBP-promoted binding of LPS to CD14, and inhibited the LBP-dependent activation of rabbit peritoneal exudate macrophages. The apparent dissociation constant for LPS-NH-LBP complexes is less than 1 x 10(-8) M which compares well with the dissociation constant for LPS-LBP complexes of approximately 1 x 10(-9) M. We conclude from these studies that the LPS binding site of LBP resides in the amino-terminal half of LBP and that the CD14 interaction site resides in the carboxyl-terminal half of LBP. These data suggest that appropriately modified fragments of LBP might provide novel reagents with high LPS binding affinity that could be useful in inhibiting LPS-dependent cellular activation in vivo.

Acute-Phase Proteins↗
PubMed1994

Vaccination of chimpanzees against infection by the hepatitis C virus.

A high incidence of community-acquired hepatitis C virus infection that can lead to the progressive development of chronic active hepatitis, liver cirrhosis, and primary hepatocellular carcinoma occurs throughout the world. A vaccine to control the spread of this agent that represents a major cause of chronic liver disease is therefore needed. Seven chimpanzees (Pan troglodytes) have been immunized with both putative envelope glycoproteins [E1 (gp33) and E2 (gp72)] that were copurified from HeLa cells infected with a recombinant vaccinia virus expression vector. Despite the induction of a weak humoral immune response to these viral glycoproteins in experimentally infected chimpanzees, a strong humoral immune response was obtained in all vaccines. The five highest responders showed complete protection against an i.v. challenge with homologous hepatitis C virus 1. The remaining two vaccines became infected, but both infection and disease may have been ameliorated in comparison with four similarly challenged control chimpanzees, all of which developed acute hepatitis and chronic infections. These results provide considerable encouragement for the eventual control of hepatitis C virus infection by vaccination.

Animals↗
PubMed1994

Effects of thyroid hormone on the calcium current and isoprenaline-induced background current in rabbit ventricular myocytes.

The majority of previous studies have been performed to explain the effects of thyroid hormone on the heart in chronic hyperthyroidism that was usually induced by eight to 10 daily injections of thyroid hormones. However, it is unclear whether or not the electrophysiological effects result from the chronic manifestations of hyperthyroidism and whether thyroid hormone acts directly or indirectly on cardiac myocytes to alter cardiac electrophysiological properties. In order to examine the acute term electrophysiological effects of thyroid hormone applied in vitro and the mechanisms responsible for some of these effects, we investigated the modulatory effects of thyroid hormone on the calcium current and isoprenaline-induced background current in L-triiodothyronine-treated ventricular myocytes of the rabbit. The major findings were as follows. Over 5 h (range, 5-24 h) after treatment of L-triiodothyronine (1 microM) in vitro, the calcium current was increased significantly. Isoprenaline (1 microM) and cyclic AMP (100 microM) caused an increase in the calcium current in both euthyroid and hyperthyroid myocytes. The hyperthyroid myocytes were more sensitive to the effect of beta-adrenergic stimulation on the calcium current and isoprenaline-activated background current. In euthyroid myocytes, acetylcholine (1 microM) produced no or little changes in the amplitude of the calcium current. In hyperthyroid myocytes, acetylcholine markedly reduced the calcium current, however, acetylcholine was ineffective in the presence of sufficient intracellular cyclic AMP (100 microM). Our results suggest that thyroid hormone can affect the cardiac myocytes directly. Furthermore, our results demonstrate that thyroid hormone affects the calcium current and isoprenaline-activated background current. These electrophysiological changes may explain, at least in part, the occurrence of positive inotropy and cardiac arrhythmias that is associated with hyperthyroidism.

Acetylcholine↗
PubMed1994

Cholecystokinin gene expression in rat amygdaloid neurons: normal distribution and effect of morphine tolerance.

Previous studies have shown that repeated opioid administration induces a tolerance to opioid, presumably due in part to an opioid-mediated compensatory increase in brain cholecystokinin (CCK) synthesis and/or release. In this study, in situ hybridization histochemistry was used to examine the effect of morphine tolerance on CCK gene expression in the amygdala of rat brains, by using a 35S-labeled synthetic oligonucleotide probe. CCK mRNA-positive neurons in normal rats were seen throughout the amygdaloid complex, with the most heavily labeled neurons in lateral, basal, and cortical nuclei, followed by the medial nucleus. Only a few labeled neurons were found in central and intercalated nuclei. The development of morphine tolerance in the rat was associated with increased hybridization signals for CCK mRNA in each subnucleus of the amygdala. Increases were seen in the numbers of positively labeled neurons and/or the numbers of hybridization grains per positively labeled neuron. Furthermore, differential patterns of increase in CCK mRNA in morphine tolerant rats occurred in different subnuclei of the amygdala, with the highest magnitude of increase in the cortical nucleus, followed in order by the medial, central, basal, intercalated and lateral nuclei. The present study demonstrated that repeated administration of morphine increased CCK gene expression in the amygdaloid complex, and suggested that the development of the tolerance to morphine analgesia is due, in part, to an increase in CCK activity in the amygdaloid complex. These findings substantiate the hypothesis that long-term administration of opioid may induce a compensatory increase in CCK synthesis and/or release, which then results in a progressive antagonism of opioid analgesia.

Amygdala↗
PubMed1994

Immunofluorescence and biochemical studies of the type VI collagen expression by human glioblastoma cells in vitro.

The human glioblastoma cell line U-87 MG was found to express a 140 kD polypeptide which was recognized on immunoblot analysis by a monoclonal antibody to type VI collagen. This polypeptide was digestible by a highly purified bacterial collagenase. After treatment of U-87 MG cells by pepsin, the protein profile revealed the two major pepsin-resistant fragments identical in Mr to those of collagen VI extracted from human placenta. The respective peptide maps from V8 protease one-dimensional gels of these two fragments were identical to those obtained with human collagen VI. Immunofluorescent staining by antibodies to type VI collagen was observed in the extracellular matrix. Moreover, U-87 MG cells were found to be positive for A2B5, a cell surface marker specific for O-2A type glial precursor cells. These data indicate that the human glioblastoma cell line U-87 MG exhibits the properties of glial precursor cells and expresses collagen type VI in vitro. This cell line therefore may prove valuable for comparative investigations of the regulation of type VI collagen synthesis, and may be useful as a model to study the function and pathological importance of type VI collagen in human brain tumours, both in vitro and in vivo.

Biomarkers↗
PubMed1994

Selection of antisense oligonucleotides on the basis of genomic frequency of the target sequence.

Antisense oligonucleotides (ASOs) are capable of blocking the expression of targeted genes and are potential antitumor and antiviral therapeutic agents. The specificity of ASO gene inhibition is compromised when homology to other sequences allows the selected ASO to bind to nontargeted mRNAs. To reduce this nonspecific activity, an ASO should target a sequence that is predicted to be unlikely to occur in other mRNAs. The probability of a sequence being unique can be predicted by determining the genomic frequency of short stretches of sequences contained within the target sequence. Two computer programs, OLIGOMER and HEXAGRAPH, were developed for this analysis. OLIGOMER was used to analyze the genomic frequencies of di-, tri-, and hexamers in more than 24 million nucleotides from 8 different genomes in GenBank. A mathematical model was developed that predicts the genomic frequency of longer oligomers on the basis of the observed frequencies of shorter oligomers. The second program, HEXAGRAPH, was used to graphically display the genomic frequency data of a selected target gene. The computational tools developed in this study may help to design more efficient ASOs by decreasing their nonspecific binding activity.

Animals↗
PubMed1994

Assessment of hepatitis C virus RNA levels by quantitative competitive RNA polymerase chain reaction: high-titer viremia correlates with advanced stage of disease.

A quantitative competitive RNA polymerase chain reaction (QC-PCR) assay was developed for measuring absolute levels of hepatitis C virus (HCV) RNA in the sera of 121 viremic persons, including 64 asymptomatic blood donors, 39 symptomatic patients referred for treatment of chronic hepatitis C, and 18 patients with end-stage liver disease referred for liver transplantation. Mean HCV RNA levels (log molecules per milliliter) were lowest among blood donors with normal alanine aminotransferase (ALT) values (5.8 +/- 1.5), higher among blood donors with elevated ALT (6.9 +/- 0.8) and clinic patients with chronic active hepatitis (6.9 +/- 0.7), and highest among patients with cirrhosis (7.1 +/- 0.8) or end-stage liver disease (7.6 +/- 1.0). High-titer viremia ( > or = 7.5 logs/mL) was more frequent among patients with end-stage liver disease (14/18; 78%) than either blood donors (10/64; P < .001) or patients with chronic active hepatitis (7/26; P < .001). Thus, 121 (94.5%) of 128 anti-HCV-positive persons were viremic. QC-PCR may be valuable for monitoring HCV infection status and selecting individuals for therapy.

Base Sequence↗
PubMed1994

Identification of residues of the H-ras protein critical for functional interaction with guanine nucleotide exchange factors.

Ras proteins are activated in vivo by guanine nucleotide exchange factors encoded by genes homologous to the CDC25 gene of Saccharomyces cerevisiae. We have taken a combined genetic and biochemical approach to probe the sites on Ras proteins important for interaction with such exchange factors and to further probe the mechanism of CDC25-catalyzed GDP-GTP exchange. Random mutagenesis coupled with genetic selection in S. cerevisiae was used to generate second-site mutations within human H-ras-ala15 which could suppress the ability of the Ala-15 substitution to block CDC25 function. We transferred these second-site suppressor mutations to normal H-ras and oncogenic H-rasVal-12 to test whether they induced a general loss of function or whether they selectively affected CDC25 interaction. Four highly selective mutations were discovered, and they affected the surface-located amino acid residues 62, 63, 67, and 69. Two lines of evidence suggested that these residues may be involved in binding to CDC25: (i) using the yeast two-hybrid system, we demonstrated that these mutants cannot bind CDC25 under conditions where the wild-type H-Ras protein can; (ii) we demonstrated that the binding to H-Ras of monoclonal antibody Y13-259, whose epitope has been mapped to residues 63, 65, 66, 67, 70, and 73, is blocked by the mouse sos1 and yeast CDC25 gene products. We also present evidence that the mechanism by which CDC25 catalyzes exchange is more involved than simply catalyzing the release of bound nucleotide and passively allowing nucleotides to rebind. Most critically, a complex of Ras and CDC25 protein, unlike free Fas protein, possesses significantly greater affinity for GTP than for GDP. Furthermore, the Ras CDC25 complex is more readily dissociated into free subunits by GTP than it is by GDP. Both of these results suggest a function for CDC25 in promoting the selective exchange of GTP for GDP.

Amino Acid Sequence↗
PubMed1994

[Report on the first nationwide survey of the distribution of human parasites in China. 1. Regional distribution of parasite species].

A nationwide (Taiwan Province not included) survey of the distribution of human parasites in China during 1988-1992 was conducted under the auspices of the Ministry of Public Health, with stratified masses randomly sampling. A total of 2,848 pilot sites in 726 counties with a population of 1,477,742 were surveyed, according to unified standard, unified diagnostic method and control quality. The overall infection rate of parasites was 62. 632%. Among them, the infection rate was over 50% in 17 provinces/autonomous regions/municipalities (P/A/M), over 80% in Hainan, Guangxi, Sichuan, Fujian, Zhejiang and Guizhou, being highest in Hainan (94. 735%). Altogether 56 species were detected. Centrocestus formosanus is reported for the first time at home, Echinochasmus liliputanus and Echinostoma angustitestis are reported for the first time at home and abroad. Echinochasmus fujianensis is a new species. E. histolytica, G. lamblia, A. lumbricoides, whipworm and pinworm were distributed nationwide, while Cysticercus (27 P/A/M), Taenia (27), hookworm (26), Balantidium coli (22), Clonorchis sinensis (22), Paragonimus westermani (21), H. diminuta (21), Echinococcus (18), H. nana (17), Fasciolopsis buski (16), T. spiralis (12) were distributed non-nationwide. A preliminary suggestion on intervention of the common and/or most detrimental parasitic diseases was submitted, including hydatidosis, taeniasis, cysticercosis, clonorchiasis, paragonimiasis, trichinellosis, hookworm disease, ascariasis, trichuriasis and enterobiasis.

Animals↗
PubMed1994

[Treatment of the squamous carcinoma of the middle ear].

This essay is based on the analysis of clinical data coming from 20 cases of squamous carcinoma of the middle ear and review of lectures on squamous carcinoma of the middle ear. It shows that no clinical factor has been formed which would affect the prognosis of squamous carcinoma of the middle ear except the involved lymph node around the primary focus. Because the radical scalpel would destroy the local anatomic structure and it is difficult to drawn the tumor tissue perfectly. The author suggest that radiotherapy is the first choice of the simple squamous carcinoma of the middle ear. The surgical and radiation after surgery is not suitable preventive clearance and radiation is not necessary for the patient whose lymph node is negative.

Adult↗
PubMed1994

[Effects of wuzi yanzong pills on lipid in rats with alcohol-induced liver injury].

Experiments showed that in the rat model of alcohol-induced liver injury, a dosage of the pills (1-2g/kg, ig) could increase the cholesterol level, lower the triglyceride level, and improve the fatty degeneration and necrosis of liver (P < 0.05-0.01). The results suggest that the important mechanism of the pills in protecting and treating alcoholic fatty liver, lies in the regulation of metabolism of the lipid, especially of the triglyceride.

Animals↗