Isolation and characterization of an IL-1-like protein from Manduca sexta.
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Animals↗
Biomedical subjects
H Zhang
Publications and source records attributed to H Zhang.
Construction and expression of eukaryotic expressing vector pCH510 of polypeptide CH50 and its chemotaxis and antitumor function by in vivo transfection.
To construct an eukaryotic expressing vector that expresses CH50, a recombinant Cell I-Hep II bifunctional-domain polypeptide of human fibronectin, and to investigate the chemotaxis to immune cells and the inhibitory effect on the growth of tumor by the expression of the plasmid in vivo, the plasmid was constructed by DNA recombination. Gene transfection was performed in vitro and in vivo. The expressed product was identified by Western blot. The chemotaxis after gene transfection in vivo was observed by histotomy and staining of muscle tissues. The inhibition of gene transfection on solid tumor was observed in mice. The results showed that plasmid pCH510 was constructed by the recombination of the 5'-terminal noncoding region and signal peptide coding region of human fibronectin cDNA and cDNA fragment coding CH50 polypeptide with a 3'-terminal noncoding region of human FN cDNA, and the insertion of the recombinated fragment into plasmid pcDNA3. 1. After transfection with plasmid pCH510, NIH3T3 cells could produce CH50 polypeptide. The transfection of plasmid pCH510 by the injection in muscle of mouse could produce the effects of chemotaxis on immune cells and the inhibition on the growth of solid tumor. It is concluded that plasmid pCH510 can express in cells and in vivo in mouse. The expression of the plasmid in vivo has a chemotactic effect on immune cells and can inhibit the growth of solid tumor.
3T3 Cells↗
A basic study on lesion detectability for hot spot imaging of positron emitters with dedicated PET and positron coincidence gamma camera.
The aim of this study was to explore the correlations of detectability and the semi-quantification for hot spot imaging with positron emitters in positron emission tomography (PET) and with a positron coincidence detection system (PCD). Phantom study results for the measurement of the lesion-to-background (L/B) ratio ranged from 2.0 to 30.3, and detectability for hot spot lesion of PET and PCD were performed to correspond to clinical conditions. The detectability and semi-quantitative evaluation of hot spots from 4.4 mm to 36.9 mm in diameter were performed from the PET and PCD images. There were strong correlations between the L/B ratios derived from PET and PCD hot spot images and actual L/B ratios; but the L/B ratio derived from PET was higher than that from PCD with a significant difference of 10% to 54.8%. The detectability of hot spot imaging of PCD was lower than that of PET at 64.8% (PCD) versus 77.8% (PET). Even the actual L/B ratio was 8.0, hot spots more than 10.6 mm in diameter could be clearly identified with PCD imaging. The same identification could be achieved with PET imaging even when the actual L/B ratio was 4.0. This detailed investigation indicated that FDG PCD yielded results comparable to FDG PET on visual analysis and semi-quantitative analysis in detecting hot spots in phantoms, but semi-quantitative analysis of the L/B ratio with FDG PCD was inferior to that with FDG PET and the detectability of PCD in smaller hot spots was significantly poor.
Gamma Cameras↗
A simple flow-injection on-line clean-up system for microwave plasma-torch atomic emission spectrometry.
A simple flow injection (FI) on-line clean-up system has been developed for microwave plasma-torch atomic emission spectrometry (MPT-AES). A non-selective strongly acidic cation-exchange resin was used to achieve the goal of "on-line clean-up". Ag and Zr, which form halogen-complex anions in halide acid media, and Cr, Mo, and P, which exist as acid group anions or acids (neutral) in acidic solution, were determined and the interfering matrix cations were removed on-line. Satisfactory analytical results were obtained from some practical samples by use of this procedure.
Journal Article↗
Long-term graft acceptance in rat heart transplantation by CTLA4Ig gene transfection combined with FTY720 treatment.
CTLA4Ig strongly adheres to B7 molecules on antigen-presenting cells to block intracellular signal transduction via CD28 on helper T cells, which eventually inhibits immune responses. We have demonstrated that the administration to recipient animals of adenoviral vectors containing CTLA4Ig gene (adCTLA4Ig) prolonged graft survival, although the gene expression diminished in a time-dependent manner and the grafts were finally rejected. In addition, recipient animals treated with FTY720, a new immunosuppressant, exhibited a decrease in the number of peripheral lymphocytes due to apoptosis. In this study, we performed adCTLA4Ig transfection combined with FTY720 treatment in heart-grafted rats to determine if the combination could induce a mutual effect on graft survival. The recipient animals were given injections of 1 x 10(9) plaque-forming units of adCTLA4Ig via the tail vein immediately after grafting. On the day before transplantation we administered FTY720 orally to some of these animals at a dosage of 5 mg/kg and again on the day of transplantation. The median graft survival period in the adCTLA4Ig-only group was 27 days, whereas that in the combination group was markedly prolonged to 56 days. Of 15 grafts, 5 survived indefinitely. In these groups we observed detectable levels of CTLA4Ig in the sera 49 days after grafting; the levels were always higher in the combination group than in the adCTLA4Ig-only group. As a result, this study revealed that FTY720 and adCTLA4Ig have a potent mutual effect on graft survival during rat heart transplantation. Furthermore, it is highly possible that FTY720 enhances gene expression of adCTLA4Ig, which may be related to the long-term acceptance of grafts.
Abatacept↗
Purification, refolding of hybrid hIFNgamma-kringle 5 expressed in Escherichia coli.
The DNA sequence coding for plasminogen kringle 5 (pK5), an inhibitor of angiogenesis, was fused with that coding for interferon gamma and over-produced in the form of inactive inclusion bodies in E. coli. The amount of fusion protein was about 40% of total protein produced. The fusion protein contained in the inclusion bodies was solubilized in 8 M urea and purified by anion-exchange chromatography. We employed the orthogonal experimental design L16(4(5)) (5 factors, 4 levels, 16 experiments) procedure for researching the influence of denaturant, aggregation suppressor L-arginine, NaCl, pH, and glycine on the refolding procedure. Our results suggest that the presence of appropriate L-arginine, NaCl, and denaturant in the refolding buffer inhibits the aggregation of the fusion protein and increases the yield of renatured protein with biological activity. The refolded fusion protein, gammaIFN/pk5, has in vitro anti-endothelial cell proliferation activity.
Angiogenesis Inhibitors↗
Cloning, molecular characterization, and application of rice epiphytic Bacillus pumilus promoter fragments.
To establish a constitutive, high-efficiency expression system for Bacillus pumilus (B.P), we cloned random chromosomal DNA into promoter probe shuttle vector ECE7 and selected for strong promoter activity by chloramphenicol resistance of transformed B. pumilus cells. The nucleotide sequences of nine chromosomal fragments were determined. These DNA fragments range from 300 to 2200 bp in size. The transcription strength of these promoters was estimated by determination of CAT enzyme production in both E. coli and B. pumilus. Transcription start (TS) sites of the cat mRNA were located by primer extension by using total RNA. Preliminary analysis showed that three of the promoter sequences contain -35 and -10 regions like E. coli RNA polymerase sigma70 and B. subtilis sigma43 consensus sequences. One is similar to B. subtilis sigma29, the other two have no conserved sequences like any of the typical consensus sequences of the known sigma factors so far. To estimate the feasibility of the utilization of these promoters, one promoter fragment was subcloned and used to drive the expression of green fluorescent protein (GFP) in B. pumilus cells. This is the first report of B. pumilus promoters randomly cloning from total DNA and molecular analysis of their consensus sequences.
Bacillus↗
An ex vivo study on radiofrequency tissue ablation: increased lesion size by using an "expandable-wet" electrode.
The present comparative study was conducted to validate a newly developed "expandable-wet" electrode for an increased lesion size of radiofrequency ablation (RFA) on excised beef liver. The expandable-wet electrode, which allows interstitial hypertonic saline infusion through retractable curved needles, was compared with "expanded-dry" and "unexpanded-wet" electrodes for RFA lesion size and other parameters. A total of 120 lesions were created under 50 W (groups A-C) and 90 W (groups A'-C') power control mode for 10 min at each ablation site with the following groups: group A and A' of expanded-dry electrode (needles deployed but saline uninfused); group B and B' of unexpanded-wet electrode (saline infused but needle undeployed); and group C and C' of expanded-wet electrode (needles deployed and saline infused). Together with lower impedance and higher power output, the lesion size in group C (5.3+/-0.4 cm) and C' (6.0+/-1.0 cm) were significantly larger (P<0.01) than that in group A (3.3+/-0.3 cm) and A' (2.0+/-0.2 cm), and group B (3.8+/-1.0 cm) and B' (2.6+/-0.4 cm). The RFA lesion size can be significantly enlarged when the expandable electrode is complemented with interstitial hypertonic saline infusion. This design may improve the efficacy of RF tumor ablation.
Animals↗
BOLD contrast fMRI of whole rodent tumour during air or carbogen breathing using echo-planar imaging at 1.5 T.
The aim of this study was to evaluate the feasibility of functional MR imaging (fMRI) at 1.5 T, exploiting blood oxygenation level-dependent (BOLD) contrast, for detecting changes in whole-tumour oxygenation induced by carbogen (5% CO2+95% O2) inhalation of the host. Adult WAG/Rij rats with rhabdomyosarcomas growing subcutaneously in the lower flank were imaged when tumours reached sizes between 1 and 11 cm3 (n=12). Air and carbogen were alternatively supplied at 2 l/min using a snout mask. Imaging was done on a 1.5-T MR scanner using a T2*-weighted gradient-echo, echo-planar imaging (GE-EPI) sequence. Analysis of the whole-tumour EPI images was based on statistical parametric maps. Voxels with and without signal intensity changes (SIC) were recorded. Significance thresholds were set at p<0.05, corrected for multiple comparisons. In continuous air breathing condition, 3 of 12 tumours showed significant negative SIC and 1 tumour had a clear-cut positive SIC. The remaining tumours showed very little or no change. When switching to carbogen breathing, the SIC were significantly positive in 10 of 12 tumours. Negative SIC were present in 4 tumours, of which three were simultaneously characterised by positive SIC. The overall analysis indicated that 6 of the 12 tumours could be considered as strong positive responders to carbogen. Our research demonstrates the applicability of fMRI GE-EPI at 1.5 T to study whole-tumour oxygenation non-invasively. The observed negative SIC during air condition may reflect the presence of transient hypoxia during these measurements. Selection of tumours on the basis of their individual response to carbogen is possible, indicating a role of such non-invasive measurements for using tailor-made treatments.
Animals↗
Spam1 (PH-20) mutations and sperm dysfunction in mice with the Rb(6.16) or Rb(6.15) translocation.
In mice bearing the Rb(6.16) or Rb(6.15) Robertsonian translocation (Rb), sperm dysfunction associated with the Rbs has been shown to lead to transmission ratio distortions (TRDs) in heterozygotes. The severity of the TRDs is directly related to the severity in the alteration of expression of the gene for the Sperm Adhesion Molecule 1 (Spam1), which maps to proximal mouse Chromosome 6 (Chr 6) near the translocation junction and encodes a sperm antigen with hyaluronidase activity. Here we demonstrate that there is a significantly reduced fertility in the Rb homozygotes (P < 0.001), based on litter size; and that with the Sperm Select Penetration assay Rb-bearing sperm have significantly decreased (P < 0.02-0.001) rates of penetration of hyaluronic acid. Catalytic kinetics studies indicate that reduced Spam1 (PH-20) hyaluronidase activity in the Rb(6.15) mice results from a qualitative defect, while for Rb(6.16) with the greater TRD both a qualitative and a quantitative deficiency (confirmed by Western analysis) of Spam1 exist. Six point mutations were shown to be clustered in the Spam1 hyaluronic acid-binding domain in Rb(6.15). For Rb(6.16) which has a gross genomic alteration at the Spam1 locus, 11 point mutations are scattered in the 5' and 3' UTRs and the coding region, where one leads to the replacement of a conserved residue. Entrapment of spontaneous Spam1 mutations, owing to recombination suppression near the Rb junctions, is proposed as the major underlying defect of the sperm dysfunction.
Animals↗
Growth suppression of hamster flank organs by topical application of catechins, alizarin, curcumin, and myristoleic acid.
Hamster flank organ growth, as measured by an increase in the area of the pigmented macule, is androgen-dependent. When flank organs of a castrated hamster are treated topically with testosterone, the flank organ becomes larger and darker. Since this growth is known to be dependent on the intracellular active androgen, 5alpha-dihydrotestosterone (DHT), inhibitors of 5alpha-reductase which converts testosterone to DHT can inhibit the growth of the flank organ. Certain unsaturated aliphatic fatty acids, such as gamma-linolenic acid and myristoleic acid, as well as other natural compounds, including alizarin and curcumin, are 5alpha-reductase inhibitors and inhibited flank organ growth. Green tea catechins, including (-)-epicatechin-3-gallate, and (-)-epigallo-catechin-3-gallate (EGCG) are also 5alpha-reductase inhibitors and inhibited flank organ growth. However, (-)-epicatechin and (-)-epigallocatechin, which are not 5alpha-reductase inhibitors, also inhibited flank organ growth. EGCG also inhibited DHT-dependent growth of flank organs. These catechins, therefore, may act by a mechanism other than inhibition of 5alpha-reductase. The effect of EGCG and other compounds was localized at the site of application; they did not affect the growth of the contralateral flank organ in the same animal. Since these compounds do not appear to exhibit systemic effects, they may be potentially useful for treatment of androgen-dependent skin disorders.
Administration, Topical↗
Effect of testosterone on Leishmania donovani infection of macrophages.
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Animals↗
Studies on the frequencies of vitamin D receptor gene polymorphism in postmenopausal women of Han and Kazak nationality in China.
Polymorphyism of the vitamin D receptor (VDR) gene is reported to play a major role in variations of the genetic regulation of bone mass. The role of VDR polymorphisms within various ethnic populations is also undetermined. The purpose of the present study was to determine the frequencies of VDR genotypes in the Han and Kazak nationalities in China. We analyzed the polymorphism defined by the Bsm1 restriction enzyme using polymerase chain reaction-restriction fragment length polymorphism in 179 healthy postmenopausal Han women and 63 healthy postmenopausal Kazak women. The genotype frequencies of the VDR were calculated later using the Hardy-Weinberg equilibrium formula. The results showed that for women of Han nationality, the bb, Bb, and BB genotypes accounted for 90.5%, 9.5%, and 0%, respectively. In Kazak women, the respective frequencies were 38.1%, 55.56%, and 6.35%. We found that there was a significant difference between women of Han and Kazak nationality in terms of the frequency of distribution of VDR genotype (P < 0.001). There was a similar distribution of VDR genotypes in Kazak women to that seen for the Caucasian population of the USA. The results of the present study provide further evidence on the different pathogeny of osteoporosis in various ethnic groups.
Bone Density↗
Radiofrequency ablation for eradication of renal tumor in a rabbit model by using a cooled-tip electrode technique.
BACKGROUND: Radiofrequency ablation (RFA) has emerged as a potential alternative for surgery in clinical oncology. This animal experiment was conducted to evaluate the feasibility of RFA in the treatment of renal tumor. METHODS: Eighteen rabbits with renal implantation of VX2 tumors were divided into two groups. Group A (n = 12) was treated with RFA by using a cooled-tip RF system at 30 W for 80 to 180 seconds. Group B (n = 6) received a sham operation. The therapeutic efficacy was evaluated by survival rate, magnetic resonance imaging (MRI), and histology. RESULTS: All animals in group B died within 3 months after tumor implantation. Total tumor eradication was achieved in 10 of 12 rabbits (83.3%) in group A, of which 5 rabbits survived longer than 6 months (absolute eradication) and another 5 rabbits were found free of viable tumor when killed (relative eradication). Two rabbits experienced local tumor relapse, lung metastasis, or both. Six-month survival rate of RFA-treated rabbits was significantly higher (P < .01) than that of control rabbits. The typical MRI appearances of the acute RFA lesion consisted of five characteristic concentric zones, which corresponded to central needle track (zone A), tumor coagulation (zone B), renal tissue coagulation (zone C), peripheral hemorrhage (zone D), and inflammatory layer (zone E) on histology. CONCLUSIONS: RFA may become a promising therapy for the treatment of renal tumor. MRI is a useful modality for assessment of renal tumor ablation.
Animals↗
Role of PTCH and p53 genes in early-onset basal cell carcinoma.
Basal cell carcinoma (BCC) is the most common skin cancer in the Western world. Ultraviolet (UV) exposure, race, age, gender, and decreased DNA repair capacity are known risk factors for the development of BCC. Of these, UVB irradiation from sunlight is the most significant risk factor. The incidence of sporadic BCC increases in individuals older than age 55, with the greatest incidence reported in individuals who are older than 70, and is rare in individuals who are younger than 30. In this study, we analyzed 24 BCC samples from individuals who had BCC diagnosed by the age of 30. Fifteen single-stranded conformation polymorphism variants in the PTCH gene were identified in 13 BCC samples. Sequence analysis of these single-stranded conformation polymorphism variants revealed 13 single nucleotide changes, one AT insertion, and one 15-bp deletion. Most of these nucleotide changes (nine of 15) were predicted to result in truncated PTCH proteins. Fifteen p53 mutations were also found in 11 of the 24 BCC samples. Thirty-three percent (five of 15) and 60% (nine of 15) of the nucleotide changes in the PTCH and p53 genes, respectively, were UV-specific C-->T and CC-->TT nucleotide changes. Our data demonstrate that the p53 and PTCH genes are both implicated in the development of early-onset BCC. The identification of UV-specific nucleotide changes in both tumor suppressor genes suggests that UV exposure is an important risk factor in early onset of BCC.
Adolescent↗
Altered expression of Bag-1 in Coxsackievirus B3 infected mouse heart.
OBJECTIVE: The mechanisms by which Coxsackie B viruses cause myocarditis or dilated cardiomyopathy are not well understood. This study examined changes in the expression of cardiac genes resulting from Coxsackievirus B3 (CVB3) infection of mice. METHODS: Mice (five per group) were experimentally infected with CVB3 or mock-infected with diluent. Altered expression of genes was initially identified by cDNA array, and confirmed by semiquantitative RT-PCR, western blot and immunohistochemistry. RESULTS: Forty-two up-regulated or down-regulated genes were observed in cDNA arrays carrying 588 known mouse genes. Among these, one down-regulated gene, Bag-1, known to be involved in inhibition of apoptosis and modulation of chaperone activity, was investigated further. Semiquantitative RT-PCR showed that Bag-1 expression was down-regulated by up to 30% in virus-infected mouse heart on day 7 compared to the mock-infected. Cell fractionation and western blot analysis confirmed that Bag-1 isoform p32 was predominant in the cytoplasm of mouse myocardium and down-regulated at 4 days or 7 days after CVB3 infection. In contrast, Bag-1 isoform p50 appeared to increase in the nuclear fraction of mouse heart at 7 days after infection. Down regulated expression and distribution of Bag-1 protein or evidence of apoptosis in the infected mouse heart was demonstrated by immunostaining or histochemistry (TUNEL assay), respectively. CONCLUSION: CVB3 infection induced differential expression of Bag-1 in cytoplasmic and nuclear fractions of mouse heart and apoptosis. This may be important in the pathogenesis of enterovirus heart muscle disease.
Animals↗
Taxane diterpenoids from the bark of Taxus yunnanensis.
Further investigation on the alcohol extract of the barks of Taxus yunnanensis led to the isolation of four taxoids, namely, 7beta-xylosyl-taxol D, taxuyunnanines P, Q and R, along with the known taxuyuntin G (2). Four are rearranged taxoids with an 11(15-->1)-abeotaxoid skeleton and an opened oxetane ring moiety. Structures were determined by spectroscopic and chemical means.
Diterpenes↗
Photochemically induced formation of the "Al13" tridecameric polycation in the presence of Fe(III) and organic acids.
We found that light-induced Fe(III) reduction associated with the oxidation of a simple hydroxy-carboxylic acid (lactate) caused the formation of the AlO4Al12(OH)24(H2O)12 polycation ("Al13"). Initial conditions were a lactate:Al:Fe ratio of 1:0.76:0.11 in a partially neutralized solution. Base was added rapidly and no Al13 was detected in samples kept in the dark. With exposure to light, Fe(III) reduction was rapid and Fe(II) reached a maximum within 1 day. After the maximum, steady-state Fe(II) declined from 54% to 43% over eight days. During this same time period, the lactate concentration fell to 2% of the original, pH rose from 4.05 to 4.46, and the Al13 detectable by 27Al NMR increased to 2.3 mmol l(-1) (51% of the total solution Al). The formation of Al13 is attributed to the pH rise resulting from the removal of the organic acid buffer. Similar photo-induced chemical changes occur in natural waters and may promote the formation of Al13, conditions permitting.
Aluminum↗