Biomedical subjects
Publications and source records attributed to W Jiang.
The decomposition rates of CaCO3 in Concha Arcae calcined at various temperatures were determined by CO2 gravimetry, and a simple method was established to judge the decomposition of CaCO3 in calcined shell drugs. In addition, the quality of shell drugs obtained commercially from six cities was compared.
To study the electrophysiologic effects of endothelin-1 (ET-1), we used patch clamp and glass microelectrode techniques to investigate the effects of ET-1 on cardiac L-Ica, Ik and Ik2 in guinea pig ventricular myocytes. The prolongation of APD50 was induced and EADs was triggered by 50 nM ET-1 perfusion. L-Ica and Ik were enhanced by various ET-1 concentration from 1 to 50 nM with dose-dependence. Their steady-state activations of L-Ica and Ik shifted left with ET-1 concentration increments. ET-1 elicited a kind of GTP-dependent inward rectifier K+ current having a mean conductance of 82.36 +/- 1.27 pS. The open time and close time (both interburst intervals and burst durations) abbreviated with ET-1 concentration increase. The results suggested that EADs -ET evoked was ascribed to the prolongation on the plateau level, which resulted from L-Ica enhancement. The ET- evoked inward rectifier K+ current should be further studied.
UNLABELLED: BACKGROUND/DESIGN: The clonal theory of cancer predicts that transformed cells within a given tumor are derived from a single initiated precursor. Advancement of this precursor through various stages of tumor development occurs with the further accumulation of selective genetic and epigenetic lesions. Mammalian ras genes are important constituents of mitogenic signaling pathways, and when activated, they contribute to deregulated cellular growth. Activated ras genes play important roles in the development of certain skin tumors. Studies on a number of animal tumor model systems have shown that ras gene activation can be an early and perhaps initial event in the development of skin tumors. Activated ras genes are also found in a significant percentage of somatic human squamous cell carcinomas. To gain retrospective insight into the stages at which activated ras genes contribute to squamous cell carcinoma development, we investigated their incidence in actinic keratoses, premalignant precursors to squamous cell carcinomas. Using a nonradioactive polymerase chain reaction-based method developed in our laboratory, we examined a panel of 19 actinic keratoses and 33 squamous cell carcinomas for activated ras genes. RESULTS: DNA analysis revealed ras gene mutations in three (16%) of 19 actinic keratoses and in four (12%) of 33 squamous cell carcinomas. Activating mutations occurred at codon 12 of the K-ras gene, and codons 12, 13, and 61 of the H-ras gene. All positive actinic keratoses and squamous cell carcinomas occurred in sun-exposed regions. CONCLUSIONS: Activated ras genes can play important roles during early stages of squamous cell carcinoma development. Aberrant repair of UV-induced pyrimidine dimers is a likely cause of this activation.
The colors, crisp extents and comminution rates of 4 kinds of shell drugs calcined under various conditions were conpared systematically. The appearance alterations of the calcined drugs were closely related to the heating degrees during calcining operation, and the effect of calcining duration was greater than that of calcining temperature.
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The internal quality of 4 kinds of shell drugs calcined under different conditions was investigated and compared in terms of the yielding ratios and total calcium content of the calcined product, Ca2+ content of the decoction and total water-soluble extractive rate. It was found that at calcining temperatures higher than 800 degrees C the internal quality of the shell drugs would change markedly.
The authors reported 20 cases of adrenocortical corcinoma. The tumors were nonfunctional in 12 cases (average age 49.7) and functional in 8 cases (5 with hypercortisolism, 3 with adrenogenital syndrome; average age 8.8). The prognosis in this group were poor, the survival of one year was 40%, two years 35% and five years 10%. Most of adrenocortical carcinomas in childhood were functional and their prognosis were poor because of delayed treatment. The nonfunctional tumors were usually diagnosed in the late period, therefore the prognosis was worse, but in recent years some cases of no secretory carcinomas had been occasionally found by image diagnosis and these tumors were small, so their prognosis was favorate.
Cyclin D1 has been implicated in G1 cell cycle progression and is frequently amplified, overtranscribed, and oversynthesized in human tumors, including esophageal carcinomas. To further address the role of cyclin D1 in cell cycle control and tumorigenesis, we have stably transfected the human cyclin D1 in the nontumorigenic esophageal epithelial cell line HET-1A. These transfected cells, which express increased amounts of cyclin D1, have enhanced colony-forming efficiency and saturation density and are resistant to growth inhibition by TGF-beta 1 compared with the parental cell line or a control vector cell clone. The clones which express increased amounts of cyclin D1 exhibited a decrease in the amount of TGF-beta type II receptor, indicating a plausible mechanism for their diminished response to TGF-beta 1. Therefore, deregulated expression of the cyclin D1 gene can modulate the negative growth factor pathway of TGF-beta 1 and may disturb the control of epithelial cell proliferation in esophageal carcinogenesis.
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Sequence-specific 1H and 15N resonance assignments have been determined for the major cold shock protein (CspA) from Escherichia coli with recently developed three-dimensional triple-resonance NMR experiments. By use of these assignments, five antiparallel beta-strands were identified from analysis of NMR data. Strands 1-4 have a classical 3-2-1-4 Greek key beta-sheet topology and there are two beta-bulges, at positions Lys10-Trp11 and Gly65-Asn66. Three-dimensional structures of CspA were generated from NMR data by using simulated annealing with molecular dynamics. The overall chain fold of CspA is a beta-barrel structure, with a tightly packed hydrophobic core. Two-dimensional isotope-edited pulsed-field gradient 15N-1H heteronuclear single-quantum coherence spectroscopy was used to characterize the 15N-1H fingerprint spectrum with and without a 24-base oligodeoxyribonucleotide, 5'-AACGGTTTGACGTACAGACCATTA-3'. Protein-DNA complex formation perturbs a subset of the amide resonances that are located mostly on one face of the CspA molecule. This portion of the CspA molecular surface includes two putative RNA-binding sequence motifs which contribute to an unusual cluster of eight surface aromatic side chains: Trp11, Phe12, Phe18, Phe20, Phe31, His33, Phe34, and Tyr42. These surface aromatic groups, and also residues Lys16, Ser44, and Lys60 located on this same face of CspA, are highly conserved in the family of CspA homologues. These isotope-edited pulsed-field gradient NMR data provide a low-resolution mapping of a DNA-binding epitope on CspA.
The major cold shock protein of Escherichia coli, CspA, produced upon a rapid downshift in growth temperature, is involved in the transcriptional regulation of at least two genes. The protein shares high homology with the nucleic acid-binding domain of the Y-box factors, a family of eukaryotic proteins involved in transcriptional and translational regulation. The crystal structure of CspA has been determined at 2-A resolution and refined to R = 0.187. CspA is composed of five antiparallel beta-strands forming a closed five-stranded beta-barrel. The three-dimensional structure of CspA is similar to that of the major cold shock protein of Bacillus subtilis, CspB, which has recently been determined at 2.45-A resolution. However, in contrast to CspB, no dimer is formed in the crystal. The surface of CspA is characteristic for a protein interacting with single-stranded nucleic acids. Due to the high homology of the bacterial cold shock proteins with the Y-box factors, E. coli CspA and B. subtilis CspB define a structural framework for the common cold shock domain.
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Escherichia coli hosts were constructed for maintenance of vectors containing the gamma replication origin of the R6K plasmid (oriRR6K gamma) at different copy numbers (15 or 250/cell). Such vectors require the trans-acting II protein (the pir gene product) for replication. New hosts carry pir+ or pir-116 on the chromosome within uidA, the E. coli gene encoding beta-glucuronidase. They were made using the rep technique for allele replacement and KmR M13 delta uid A::pir+ or M13 delta uidA::pir-116 phage. Because M13 cannot replicate in a rep mutant, KmR transductants arose by integration into the chromosomal uidA locus. Segregants lacking M13 sequences (which were selected as deoxycholate-resistant (DocR) ones) frequently contained delta uidA::pir+ or delta uidA::pir-116 on the chromosome. In principle, this procedure could be used for the introduction of any foreign gene into any nonessential gene on the E. coli chromosome. The delta uidA::pir+ and delta uidA::pir-116 loci were subsequently transferred to a variety of E. coli strains. One such strain is a suppressor-negative one that is especially useful for transposon (Tn) mutagenesis. This strain has an integrated RP4 derivative for conjugative transfer of oriRR6K gamma plasmids also containing oriT from RP4. In addition, new oriRR6K gamma, oriT+ vectors carrying the TcR-encoding genes tetAR from Tn10 are described. These can be used for allele replacement by conjugative transfer of an oriRR6K gamma, oriT+, tetAR plasmid containing a mutated gene into a non-pir recipient and by subsequent selection for Tc-sensitive exconjugants.
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We previously isolated a partial cDNA sequence, termed TPAR1 (TPA repressed gene 1), from a cDNA library constructed from C3H10T1/2 mouse embryo fibroblasts treated with TPA, using a differential screening procedure. (M.D. Johnson et al. Mol. Cell. Biol. 7, 2821-2829, 1987). In the present study, we have cloned two corresponding full-length 1.9- and 3.4-kb cDNAs of TPAR1 from murine cDNA libraries. Sequence analysis of these TPAR1 cDNAs revealed that they encode 89 and 93 amino acid polypeptides, respectively, with a putative leader sequence and show significant homology with the human cytokine interleukin-8 (IL-8) and its superfamily. Genomic DNA isolation and structural characterization provide evidence that the TPAR1 mRNAs are transcribed from a single gene with alternative splicing. TPAR1 mRNAs are expressed ubiquitously among adult mouse tissues as three major transcripts, 1.9, 3.4, and 6.5 kb, whose expression depends on the tissue type. The levels of TPAR1 mRNAs were markedly decreased in fibroblasts following TPA treatment and also in serum-deprived quiescent fibroblasts stimulated by serum. The levels of TPAR1 mRNAs were dramatically down-regulated in regenerating rat liver when compared to normal adult liver. In addition, there was no detectable expression of TPAR1 in three rat hepatoma cell lines and several transformed fibroblast cell lines. Thus, the TPAR1 gene is a new member of the cytokine IL-8 superfamily, whose expression is down-regulated in rapidly dividing cells. Further studies are required to determine whether it plays a negative role in controlling cell proliferation and tumorigenesis.
A recombinant single chain antibody fragment (scFv) that identifies a neutralizing epitope on the envelope glycoprotein of louping iII (LI) and tick-borne encephalitis (TBE) virus has been developed using a bacteriophage expression system. The mRNA was extracted from a cloned hybridoma cell culture that produces a mouse monoclonal antibody (MAb 4.2) known to map to amino acids 308-311 of LI and TBE virus, corresponding to domain B on the proposed two-dimensional model of the tick-borne encephalitis virus envelope protein. The V-genes encoding the antigen-binding site of MAb 4.2 were amplified and cloned for expression as a fusion protein to the pIII coat protein of filamentous phage. Solid phase selection of these phage against the LI virus antigen, was necessary to isolate the correct MAb 4.2 scFv fragment which was subsequently produced in soluble form in bacteria and harvested from the culture supernatant medium. The characteristics of this expressed single chain antibody were compared with MAb 4.2. The expressed antibody portrayed the antigenic specificity of MAb 4.2 and also neutralized the infectivity of louping iII and some other tick-borne flaviviruses. The potential of this technique for studying antigen-antibody interactions and for the development of prophylactic reagents are discussed.
The effects of different concentrations of lead nitrate on root growth, cell division, chromosome morphology and nucleolus in root tip cells of Allium cepa were studied. The concentrations of lead nitrate (Pb(NO(3))(2)) were 10(-7), 10(-6), 10(-5), 10(-4), 10(-3) and 10(-2) m. It was observed that lead reduces root growth and causes mitotic irregularities, including c-mitosis, anaphase bridges, and chromosome stickiness. Also, interphase cells with micronuclei, irregularly shaped nuclei and nuclei with decomposed nuclear material were noticed. The c-mitotic effect reached its maximum in the meristem at above 10(-4) m Pb, when practically almost all the anomalous dividing cells are of this type. After treatment with Pb, there were many similar silver-stained particulate materials scattered in the nucleus in short, rounded meristem cells and in long, oblong root cap cells.