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Biomedical subjects

C Shi

Publications and source records attributed to C Shi.

At least 19 recordsLinked to original sources
PubMed1997

Hypercholesterolemia exacerbates transplant arteriosclerosis via increased neointimal smooth muscle cell accumulation: studies in apolipoprotein E knockout mice.

BACKGROUND: Hypercholesterolemia is thought to be a significant risk factor for coronary vasculopathy in cardiac transplant recipients. METHODS AND RESULTS: We examined the development of arteriosclerosis in mouse carotid artery loops allografted from B.10A(2R) (H-2h2) donors to normocholesterolemic C57BL/6J (H-2h) recipients and hypercholesterolemic C57BL/6J recipients in which the apolipoprotein (apo) E gene had been knocked out. Luminal occlusion and cross-sectional neointimal area were greater in arteries allografted into hypercholesterolemic recipients at 15 and 30 days after transplantation. We also measured cellular and extracellular matrix components of the neointima by computerized planimetry of the fractional areas subtended by smooth muscle cells (anti-alpha-actin stain), collagen (Masson's trichrome), lipid (oil red O), and leukocytes (anti-CD45). The neointimal area stained for smooth muscle cells was significantly greater in hypercholesterolemic recipients than in normocholesterolemic recipients at 15 and 30 days after allografting. Lipid contributed to neointimal area to a lesser degree, and there was no significant increase in the contribution of collagen or leukocytes. CONCLUSIONS: Smooth muscle cell accumulation appears to be the principal contributor to the increase in neointimal area observed in arteries allografted into hypercholesterolemic mice.

Animals↗
PubMed1997

A recombinant retroviral system for rapid in vivo analysis of human immunodeficiency virus type 1 susceptibility to reverse transcriptase inhibitors.

We have developed a new recombinant retroviral system in which a library of infectious molecular clones of human immunodeficiency virus type 1 (HIV-1) is constructed with reverse transcriptase (RT) genes derived from viral RNA sequences in plasma. HIV-1 RT is amplified from plasma HIV-1 RNA by nested RT-PCR and cloned into a RT-defective HIV-1 proviral vector (xxLAI-np), generating 10(3) to 10(4) recombinant proviral clones from each reaction. The bulk cloning products or individual molecular clones are transfected into MT-2 cells to generate infectious virus. The resultant viruses are assayed for drug susceptibility in CD4+ cell lines to determine either the dominant phenotype of the recombinant virus mixture or the phenotypes of the individual viral clones. DNA sequencing of the cloned RT genes can identify mutations associated with phenotypic resistance of clonal mixtures or individual clones. This method can be used to rapidly detect the in vivo emergence of HIV-1 quasispecies resistant to RT inhibitors.

Anti-HIV Agents↗
PubMed1997

[A promoter responsible for over-expression of cholera toxin B subunit in cholera toxin A subunit structure gene].

A promoter sequence, which promotes the transcription of cholera toxin B subunit gene, was found in cholera toxin A subunit structure gene. The transcription starts at the adenine Located at +833, that is 456bp upstream to the A of the initiation codon ATG of cholera toxin B gene. Under the control of the promoter, cholera toxin B subunit was over-expressed as high as 200 mg/L at an optimized culture condition. The chloramphenicol acetyl transferase gene and beta-galactosidase could also be efficiently expressed under the direction of the promoter. This promoter may be responsible for the 6 fold and 7 fold higher expression level of cholera toxin B subunit than cholera toxin A subunit in V. cholerae and Escheria coli respectively. The over-expression of CTB may be useful in preparing vaccine against cholera and facilitating the construction of peptide-bearing immunogenic hybrid proteins.

Base Sequence↗
PubMed1996

Immunologic basis of transplant-associated arteriosclerosis.

Although immunosuppressive therapy minimizes the risk of graft failure due to acute rejection, transplant-associated arteriosclerosis of the coronary arteries remains a significant obstacle to the long-term survival of heart transplant recipients. The participation of specific inflammatory cell types in the genesis of this lesion was examined in a mouse model in which carotid arteries were transplanted across multiple histocompatibility barriers into seven mutant strains with immunologic defects. An acquired immune response--with the participation of CD4+ (helper) T cells, humoral antibody, and macrophages--was essential to the development of the concentric neointimal proliferation and luminal narrowing characteristic of transplant arteriosclerosis. CD8+ (cytotoxic) T cells and natural killer cells were not involved in the process. Arteries allografted into mice deficient in both T-cell receptors and humoral antibody showed almost no neointimal proliferation, whereas those grafted into mice deficient only in helper T cells, humoral antibody, or macrophages developed small neointimas. These small neointimas and the large neointimas of arteries grafted into control animals contained a similar number of inflammatory cells; however, smooth muscle cell number and collagen deposition were diminished in the small neointimas. Also, the degree of inflammatory reaction in the adventitia did not correlate with the size of the neointima. Thus, the reduction in neointimal size in arteries allografted into mice deficient in helper T cells, humoral antibody, or macrophages may be accounted for by a decrease in smooth muscle cell migration or proliferation.

Animals↗
PubMed1996

Post-transcriptional stimulation of transforming growth factor beta 1 mRNA by TGF-beta 1 treatment of transformed human osteoblasts.

Following exogenous administration of transforming growth factor-beta 1 (TGF-beta 1) polypeptide to the human osteosarcoma cell line TE-85, we observed a 2- to 6-fold stimulation of steady-state TGF-beta 1 mRNA. The stimulation was dose- and time-dependent, as judged from Northern blot hybridization analyses. A 2- to 6-fold increase of the TGF-beta 1 polypeptide was also found in the media of these cells after TGF-beta 1 treatments. The autostimulation of TGF-beta 1 mRNA was nullified by cycloheximide treatment of the cells. The in vitro transcription rates of the TGF-beta 1 gene by isolated nuclei were not altered by TGF-beta 1 treatment. Under conditions of transcriptional inhibition, the stability of TGF-beta 1 mRNA was enhanced nearly two-fold by TGF-beta 1 treatment. Our findings indicate that TGF-beta 1 can stimulate autologous gene expression and subsequent polypeptide translation by a post-transcriptional mechanism requiring protein synthesis in human osteoblast-like cells. The recognized versatility of TGF-beta 1 autostimulation mechanisms (transcriptional and post-transcriptional) in other mesenchymal cells may apply also to skeletal cells, further underscoring the broad and potent activities of this cytokine.

Cell Line, Transformed↗
PubMed1996

Rapid-processing procedure for heat polymerization of polymethyl methacrylate in a pressure cooker with automatic controls.

This study introduces a new rapid-processing procedure for curing polymethyl methacrylate denture base resin in an automatically controlled pressure cooker. The cooker filled with water was inflated with 6 kgf/cm2 air pressure and heated to 120 degrees C (248 degrees F) and maintained for 10 minutes. No significant differences were found between the new pressure cooker method and the conventional method for surface hardness and porosity (p > 0.05). The pressure cooker significantly shortened polymerization time, and the polymerization is controlled automatically.

Air Pressure↗
PubMed1996

[Detection of expression of multidrug resistance gene in breast cancer by RT-PCR].

Using RT-PCR technique, we established a method for measuring expression of mdr-1 gene in breast cancer and the conditions affecting PCR results. Expression of mdr-1 gene was detected in 74 breast cancer samples. The results showed that 34.3%(12/35) untreated primary lesions and 59.0%(23/39) relapsed metastatic lesions had positive expression of mdr-1 gene (P < 0.05). In 32 patients, the overall correlation of mdr-1 gene expression with clinical drug sensitivity was 78.1%. The results validate that RT-PCR assay is highly sensitive, relatively quantitative and reliable for measuring levels of mdr-1 gene expression in clinical samples.

Breast Neoplasms↗
PubMed1996

[An experimental and clinical study on radix Salviae miltiorrhiae in the treatment of hepatocellular Ca2+ overload during hepatic ischemia/reperfusion injury].

In recent years there has been considerable interest concerning the role of hepatocellular Ca2+ overload which probably was a major factor in hepatic ischemia/reperfusion injury. We studied the effect of radix salviae miltiorrhizae (RSM) on cytosolic free calciumion concentration [(Ca2+)i](nM) in isolated hepatocytes in rats and patients with ischemia reperfusion by microflurometry using fluorescent (Ca2+)i indicator Fura-2/AM. Changes of lipid peroxide free radical (ROO.) signal ranges with in the liver tissue by ESR technique and those of hepatocellular ultrastructure by electronmicroscope were also observed. The results showed that RSM reduced levels of (Ca2+)i and ROO. Ymax (mm) ESR signal rangs. RSM had an effect on protecting hepatocytes against ischemia/reperfusion injury as a useful receptor-operated calcium channels (ROC) blocker.

Animals↗
PubMed1995

LEW-to-F344 carotid artery allografts: analysis of a rat model of posttransplant vascular injury involving cell-mediated and humoral responses.

A key manifestation of chronic rejection is an obliterative arteriosclerosis. Myointimal thickening in the vessel is preceded by an endothelialitis involving accumulation of host mononuclear cells in the perivascular and intimal spaces. We report a paratopic LEW-to-F344 rat carotid artery transplantation model developed to study the cells, cytokines, and inflammatory response associated with this early phase of vascular immune injury. Compared with contralateral control arteries and isografts, LEW-to-F344 carotid allografts develop intimal thickening with mononuclear cell infiltration that persists (days 20, 45, 75, 90, and 120). Allografted vessels had dense collections of intimal and adventitial leukocytes (CD45+) consisting of equal numbers of T cells and macrophages. There were small but variable numbers of intimal smooth muscle cells. Intimal cells showed dense staining for tumor necrosis factor-alpha, interleukin-8, platelet-derived growth factor, iNOS, and ICAM and weaker labeling for interleukin-1 beta and interleukin-6. There was also prominent staining for interleukin-4 and interleukin-7 with no detectable interferon-gamma or interleukin-2 staining and high titer labeling for IgG1 (but not IgG2). The predominance of the T cell infiltrate coupled with interleukin-4 and IgG1 expression in carotid allografts is consistent with a TH2 response. This contrasts with balloon-injured rat carotids, which evoke a macrophage-dependent proliferative response. These findings demonstrate that there are distinct as well as common activation pathways in various forms of vascular injury and the LEW-to-F344 carotid model provides the opportunity to gain insight into molecular mechanisms regulating alloimmune injury in the vessel.

Animals↗
PubMed1995

Nucleotide sequence of maize dwarf mosaic virus capsid protein gene and its expression in Escherichia coli.

The 3'-terminal 1,279 nucleotide sequence of maize dwarf mosaic virus (MDMV) genome has been determined. This sequence contains an open reading frame of 1,023 nucleotides and a 3'-non-coding region of 256 nucleotides. The open reading frame includes all of the coding regions for the viral capsid protein (CP) and part of the viral nuclear inclusion protein (NIb). The predicted viral CP consists of 313 amino acid residues with a calculated molecular weight of 35,400. The amino acid sequence of the viral CP derived from MDMV cDNA shows about 47%-54% homology to that of 4 other potyviruses. The viral CP gene was constructed in frame with the lacZ gene in pUC19 plasmid and expressed in E. coli cells. The fusion polypeptide positively reacted in Western blot with an antiserum prepared against the native viral CP.

Amino Acid Sequence↗
PubMed1995

A secretion expression system using promoter and signal peptide of cholera toxin B subunit gene.

A secretion expression plasmid vector pMC05S was constructed taking advantage of the promoter, signal peptide, and transcriptional terminator of cholera toxin B subunit gene and beta-galactosidase was overexpressed in E. coli and most of the expressed enzyme was secreted into periplasma when the lacZ gene was inserted downstream of the signal peptide sequence of pMC05S. The yield of beta-galactosidase by engineered E. coli reached 30 mg/L and most of the beta-galactosidase retained the activity of the enzyme. The appropriate host strain and medium were also investigated. This system provided a new approach for the expression of proteins that easily form inclusion bodies.

Base Sequence↗
PubMed1995

Chinese character operating system of traditional Chinese medicine and pharmacology (TCMP).

With the development in research, teaching and literature work in traditional Chinese medicine and pharmacology (TCMP) by means of computers, it has been found that the existing Chinese character operative systems cannot meet the need of carrying out information processing and software development in this field, since these systems do not include many of the common and special terms in TCMP. This makes it inconvenient to exchange academic thoughts in information processing in this field with our colleagues at home and abroad and greatly affects the sharing of the literature data in TCMP. It is therefore necessary to develop a Chinese character operating system applicable to the use of computers in the research of TCMP. Recently, we have developed jointly a Chinese character operating system of TCMP. This system is based on the original GB2312-80 Chinese character international code, to which are added 1,150 Chinese characters commonly used in TCMP. The five-stroke code, which make input possible according to word forms, are used for the input of expanded words. Besides, the system also provides the codes of a number of common names of Chinese materia medica, acupoint names, common terms in traditional Chinese medicine (TCM), TCM disease names, names of classic TCM works, etc. It also provides a convenient character-creating software. We hope that our work will lead to discussions concerning the difficult problems in computer processing of TCMP literature, i.e. the Chinese character.

Database Management Systems↗
PubMed1995

[Secretory immunoglobulin A in human milk and infants' feces at 1-4 months after delivery].

OBJECTIVE: To determine the concentration of secretory immunoglobulin A (SIgA) in human milk at 1, 2, 3 and 4 months after delivery and in infants' feces in the same period and find out the effect of milk on the content of SIgA in infants' intestine. METHODS: 20 women and 12 breast feeding infants and 13 formula feeding infants were studied, and milk and infants' feces were collected at 1, 2, 3 and 4 months after delivery, respectively. The concentrations of SIgA in milk and feces were determined by radioimmunoassay. RESULTS: No apparent changes in the concentration of SIgA were found in human milk and feces from breast feeding and formula feeding infants at 1-4 months after delivery, but the concentration of fecal SIgA in breast feeding infants was significantly higher than that in the formula feeding infants, during the first 4 months after birth. CONCLUSIONS: Human milk can provide a large amount of SIgA for infants. We conclude from these data that human milk is beneficial in protecting the immunological function of infants digestive tract.

Breast Feeding↗
PubMed1994

Regeneration of HoxD expression domains during pattern regulation in chick wing buds.

The expression domains of genes located at the 5' end of the HoxD (formerly Hox-4) complex appear to correlate with pattern along both the proximal-distal (PrDi) and the anterior-posterior (AP) axes of the developing limb bud, and it has been suggested that the HoxD gene products are involved in the specification of positional information during limb development. The apical ectodermal ridge is required for limb outgrowth and is thought to influence mesodermal cells at the distal end of the limb bud in a region within which patterning events occur. In this paper, we examine the expression of 5' HoxD genes during PrDi pattern regulation in chick wing buds. In limbs undergoing pattern regulation, we demonstrate that the domains of HoxD11 and HoxD13 gene expression are "regenerated" within 24 hr of removal of the distal mesenchyme. In contrast, in limbs which will not form distal structures, HoxD13 expression becomes reduced.

Animals↗
PubMed1994

Resistance of human immunodeficiency virus type 1 to acyclic 6-phenylselenenyl- and 6-phenylthiopyrimidines.

Acyclic 6-phenylselenenyl- and 6-phenylthiopyrimidine derivatives are potent and specific inhibitors of human immunodeficiency virus type 1 (HIV-1). The development of in vitro resistance to two derivatives, 5-ethyl-1-(ethoxymethyl)-(6-phenylthio)-uracil (E-EPU), was evaluated by serial passage of HIV-1 in increasing concentrations of inhibitor. HIV-1 variants exhibiting > 500-fold resistance to E-EPSeU and E-EPU were isolated after sequential passage in 1, 5, and 10 microM inhibitor. The resistant variants exhibited coresistance to related acyclic 6-substituted pyrimidines and the HIV-1-specific inhibitors (+)-(5S)-4,5,6,7-tetrahydro-5- pyrimidines and the HIV-1-specific inhibitors (+)-(5S)-4,5,6,7-tetrahydro-5- methyl-6-(3-methyl-2-butenyl)imidazo[4,5,1-jk]benzodiazepin-2(1H)- thione (TIBO R82150) and nevirapine, but remained susceptible to 3'-azido-3'-deoxythymidine, 2',3'-dideoxycytidine, 2',3'-dideoxyinosine, and phosphonoformic acid. DNA sequence analysis of reverse transcriptase (RT) derived from E-EPSeU-resistant virus identified a Tyr (TAT)-to-Cys (TGT) mutation at either codon 188 (Cys-188; 9 of 15 clones) or codon 181 (Cys-181; 5 of 15 clones). The same amino acid changes were found in RT from E-EPU-resistant virus, but the Cys-181 mutation was more common (9 of 10 clones) than the Cys-188 mutation (1 of 10 clones). Site-specific mutagenesis and production of mutant recombinant viruses demonstrated that both the Cys-181 and Cys-188 mutations cause resistance to E-EPSeU and E-EPU. Of the two mutations, the Cys-188 substitution produced greater E-EPSeU and E-EPU resistance. The predominance of the Cys-188 mutation in E-EPSeU-resistant variants has not been noted for other classes of HIV-1 specific RT inhibitors. HIV-1 resistance is likely to limit the therapeutic efficacy of acyclic 6-substituted pyrimidines if they are used as monotherapy.

Antiviral Agents↗
PubMed1994

Murine model of accelerated transplant arteriosclerosis.

To define the role of specific gene deletions and mutations in the development of transplant arteriosclerosis, we generated an accelerated model of the disease in mice. Carotid arteries were transplanted between B.10A(2R) (H-2h2) donor mice and C57BL/6J (H-2b) recipients and compared with arteries isografted between H-2b mice. Immunosuppressive drugs were not used. Within 7 days, the allografted carotid artery formed a neointima composed of mononuclear leukocytes (CD45+) that were predominantly monocytes or macrophages (ie, CD11b+ cells with single-lobed nuclei). CD4+ and CD8+ cells were present as well. By 30 days, the neointima became exuberant, and mononuclear leukocytes were largely replaced by smooth muscle cells. Cells staining for proliferating-cell nuclear antigen were abundantly present in the intima at both early and late time points, indicating the proliferation of mononuclear leukocytes and smooth muscle cells. The area of the intima increased from day 7 to day 30 (P < .0005), as did the number of nuclei (P = .0005), but the density of the nuclei decreased (P = .02), suggesting the formation of extracellular matrix. Six of the eight isografts formed no neointima, and in samples from the remaining two, a single layer of smooth muscle neointimal cells covered just a portion of the vessel circumference. This model, which reproduces many of the features of human transplant arteriosclerosis but at an accelerated pace, should prove useful for determining the roles in transplant arteriosclerosis of genes that code for components of immunologic and inflammatory responses.

Animals↗
PubMed1994

[The upstream sequence of cholera toxin B subunit gene: effect on CTB expression].

In this work, we have studied the effect of cholera toxin A structure gene on the expression of the distal ctxB gene by the methods of deletion and frame-shift mutation. The results showed that: The expression level of Plasmid pUC19CTB, which was constructed by cloning the XbaI-EcoRI restriction fragment into pUC19 and ctxA gene was out-frame with lacZ' gene, is about 30 micrograms/ml; If a frame shift mutation was introduced at XbaI site of pUC19CTB so that the cholera toxin A gene was inframe with lacZ' and could be translated, the expression level of ctxB was decreased to 12 micrograms/ml; When A further deletion from XbaI to ClaI of cholera toxin A gene (about 550bp) was made and ctxA was outframe with LacZ', ctxB expression was decreased two fold compared to pUC19CTB; If the ctxA was inframe with LacZ' so ctxA gene could be translated, the expression level of CTB is much lower than the plasmid outframe with lacZ'. These observations could not be explained by the current knowledge about genetical regulation of cholera toxin operon. The promoter we found located in the cholera toxin A subunit gene, which is responsible for the expression of cholera toxin B subunit, may answer the question why the 550bp non-coding sequence could enhance the expression of cholera toxin B subunit.

Base Sequence↗