[Progress in the research on the tachyzoite-bradyzoite conversion of Toxoplasma gondii].
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Animals↗
Biomedical subjects
X G Chen
Publications and source records attributed to X G Chen.
[Purification of soluble egg antigen (SEA) 38 kDa molecule of Schistosoma japonicum and preliminary evaluation of its immunodiagnostic efficacy].
OBJECTIVE: To study the applicability of soluble egg antigen (SEA) 38 kDa molecule of Schistosoma japonicum in the immunodiagnosis of schistosomiasis. METHODS: The target immunodiagnositic antigen SEA 38 kDa molecule was separated and purified by SDS-PAGE and electroelution technique and evaluated by SDS-PAGE and Western blotting. To compare their immunodiagnostic efficacy, the purified SEA 38 kDa and SEA were used to examine the sera from 182 cases including 31 acute schistosomiasis patients, 31 chronic schistosomiasis patients, 60 normal human controls, 30 paragonimiasis patients and 30 clonorchiasis patients. RESULTS: The positive rates of purified SEA 38 kDa and SEA for acute schistosomiasis patients, chronic schistosomiasis patients, normal human controls, paragonimiasis patients and clonorchiasis patients were 90.3%, 90.3%, 1.7%, 0, 0 and 90.3%, 83.9%, 3.3%, 0, 0, respectively. The P/N ratio of purified SEA 38 kDa and SEA in immunodiagnosis of schistosomiasis by ELISA had been compared by two-sample t-test. It was demonstrated that there existed significant differences between purified SEA 38 kDa and SEA in the three groups of acute and chronic schistosomiasis patients and normal persons(P < 0.001). The P/N ratio of SEA 38 kDa in detecting the sera from schistosomiasis patients was obviously higher than that of SEA, but when SEA 38 kDa was used to examine normal human sera, the result was converse. CONCLUSION: The purified SEA 38 kDa has higher sensitivity and specificity and immunodiagnostic potential.
Animals↗
Effects of genistein, a soybean-derived isoflavone, on proliferation and differentiation of B16-BL6 mouse melanoma cells.
Genistein, a soybean-derived isoflavone, may contribute to the lower cancer incidence in South Asian countries. In this study, the effects and molecular mechanisms of genistein on growth and differentiation of B16-BL6 mouse melanoma cells were investigated. Genistein suppressed the growth of these melanoma cells. The IC50 value is 15.5 microM. On the other hand, genistein induced the changes of cell shape and cytoskeletal network. The cytoskeletal filaments were induced to form a bundle along the direction of elongation of the cells. Moreover, tyrosine phosphorylation levels of cytoskeleton-associated proteins decreased after the cells were exposed to 20 or 30 microM of genistein for 3 days. All these morphological and molecular changes were accompanied by appearance of the differentiated phenotypes. Genistein induced the increase of cellular melanin content, enhancement of tyrosinase activity, and decrease of colonization potentials in soft agar in a time-dependent and dose-dependent manner. The effective concentration was no more than 10 microM after 3 days' exposure. The tumorigenic potentials of B16-BL6 cells in C57BL/6 mouse also decreased after exposure to 20 or 30 microM of genistein for 3 days. When expressions of tumor-related genes were investigated in the differentiation-induced cells, the content of P53 dramatically increased while that of c-Myc protein decreased. Therefore, due to its ability to induce cellular and molecular changes, genistein suppressed the growth and induced differentiated phenotypes in B16-BL6 melanoma cells.
Animals↗
Baculovirus expression of the major surface antigen of Toxoplasma gondii and the immune response of mice injected with the recombinant P30.
The major surface antigen (P30) of the Toxoplasma gondii was expressed by an insect cell culture system infected with recombinant baculovirus. About 750 microg of purified (95% purity) P30 was obtained from a culture of 10(8) insect Sf21 cells. The recombinant P30 was used to immunize mice to induce immune response. Mice injected with the recombinant protein produced specific humoral and cellular immune responses. Immunization with P30 also prolonged the period of survival of mice infected by Toxoplasma. The average survival time of control group is 13.25+/-1.16 days, but are 16.13+/-2.1 days, 19.50+/-3.21 days, 20.38+/-3.38 days in different immunized groups, respectively.
Animals↗
PCR cloning and expression of the molt-inhibiting hormone gene for the crab (Charybdis feriatus).
A PCR-based genomic DNA walking technique was used to clone the gene for the molt-inhibiting hormone of the crab, Charybdis feriatus. Several overlapping genomic clones were isolated, and the MIH gene for the crab was reconstructed. DNA sequence determination of the overlapping clone reveals that the MIH gene spans 4.3kb and consists of three exons and two introns. Exons 1 and 2 carry a coding sequence for the signal peptide, and exons 2 and 3 consist of coding sequence for the mature peptide. The exon-intron boundary of the crab MIH gene also follows the 'GT-AG rule' for the splice donor and acceptor. The deduced amino acid sequence of MIH shows the highest overall similarity to those of the crabs, Callinectes sapidus and Carcinus maenas, and the gonad-inhibiting hormone (GIH) of the lobster. The putative polyadenylation signal is approximately 1.0kb 3' downstream of the termination codon (TGA). Genomic Southern blot analysis indicates that few genomic fragments were hybridized to the cDNA probe. The 5' flanking region contains a putative promoter with several putative cis elements similar to some vertebrate neuropeptide genes. The 530-bp flanking region was subcloned separately to two promoterless reporter plasmids carrying either the Green Fluorescent Protein gene (GFP) or the Choramphenicol Acetyltransferase gene (CAT). The DNA constructs were transfected into insect cells (Sf21) and mouse pituitary cells (GH4ZR7), respectively. Green fluorescent protein was detected in some of the transfected insect cells, and expression of the CAT was detected in cells transfected with DNA constructs containing the crab promoter. By RT-PCR, MIH transcripts can be detected in the eyestalk of shrimp in intermolt, early premolt, late premolt stages and females that brood their eggs. It can also be found in the brain, but not in the ovary, hepatopancreas, muscle and epidermis. During early larval development, MIH mRNA can be detected in the pre-hatched and the newly hatched larvae. Unlike the adult, the expression of the MIH in the larvae is exclusively in the brain.
Amino Acid Sequence↗
UV resonance Raman-selective amide vibrational enhancement: quantitative methodology for determining protein secondary structure.
We have directly determined the amide band resonance Raman spectra of the "average" pure alpha-helix, beta-sheet, and unordered secondary structures by exciting within the amide pi-->pi* transitions at 206.5 nm. The Raman spectra are dominated by the amide bands of the peptide backbone. We have empirically determined the average pure alpha-helix, beta-sheet, and unordered resonance Raman spectra from the amide resonance Raman spectra of 13 proteins with well-known X-ray crystal structures. We demonstrate that we can simultaneously utilize the amide I, II, and III bands and the Calpha-H amide bending vibrations of these average secondary structure spectra to directly determine protein secondary structure. The UV Raman method appears to be complementary, and in some cases superior, to the existing methods, such as CD, VCD, and absorption spectroscopy. In addition, the spectra are immune to the light-scattering artifacts that plague CD, VCD, and IR absorption measurements. Thus, it will be possible to examine proteins in micelles and other scattering media.
Amides↗
[The diversity of human hematopoietic stem/progenitor cells: IX. Cell-cycles kinetics related macromolecules DNA, RNA and protein contents in CD34+ hematopoietic cells of human bone marrow].
DNA, RNA and PRO comprise the bulk of macromolecules in cells, which have been proven to play an important role in regulating cell cycle transverse capacity, cell division, growth, and size. Simultaneous analysis of these moieties could provide more comprehensive and accurate information on cell cycle kinetics. In this study, DNA, RNA and PRO contents related to cell cycle kinetics in CD34+ hematopoietic cells of human bone marrow were measured to understanding the cell cycle kinetic features in CD34+ hematopoietic cells. For this reason, CIMS-100 immunomagnetic isolator, a novel isolation system, was used to enrich efficiently CD34+ hematopoietic cells from human bone marrow. The purity of enriched CD34+ hematopoietic cells determined by both FACS and APAAP staining is ranging from 90%-95%. Cellular DNA, RNA and PRO were stained with fluorochromes propidium iodide, pyronin Y and fluorescein isothiocyante respectively The fluorescence intensities reflecting the DNA, RNA and PRO content of individual cell were analyzed in FACS can by different excitation wavelengthes. DNA, RNA and PRO contents in CD34+ hematopoietic cells were far lower than these of bone marrow mononuclear cells, only being 34 +/- 3% (DNA), 48 +/- 21% (RNA) and 62 +/- 14% (PRO) of BMMNCs respectively. Collectively, these data combined with previous results from both ours and others indicated that CD34+ hematopoietic cells are indeed an unique cell population, not only in reconstitute of hematopoietic and immunological functions, but also in cell cycle kinetics. This is, to our knowledge, the first detailed report on the analysis of DNA, RNA and PRO contents related to cell cycle kinetics in CD34+ hematopoietic cells. And the results provide more direct evidence that the majority of CD34+ hematopoietic cells are in resting state.
Antigens, CD34↗
Simultaneous assessment of conformation and aggregation of beta-amyloid peptide using electrospray ionization mass spectrometry.
Electrospray ionization mass spectrometry was used to study conformation and aggregation of the synthetic beta-amyloid peptide, residues 1-40 (betaA4), as a function of concentration and sample aging. All mass spectra showed a major envelope of peaks corresponding to charge states of 7-3 of the monomeric form of betaA4. In addition, weaker envelopes of peaks corresponding to charge states of dimeric, trimeric, and tetrameric betaA4 species were seen under gentle ionization conditions. The average charge state of the envelope associated with the monomeric form decreased by ca. 0.5 z as samples were aged, indicating that the relatively open form (likely random coil) of the peptide was modified into the more compact form (likely beta-sheet) as a function of sample aging. The aggregate forms became weaker and ultimately were absent both in the more dilute solutions and in aged aliquots of the concentrated sample. These aggregates were interpreted as assemblies of the random coil form. We interpret our inability to see an ion envelope that can be associated with aggregates of the beta-sheet form to be a consequence of the presumed very compact nature of this form. A model for the formation of betaA4 fibrils is proposed and discussed.
Amyloid beta-Peptides↗
The infection status of anisakid larvae in marine fish and cephalopods from the Bohai Sea, China and their taxonomical consideration.
The infection status of anisakid larvae was examined in 290 marine fish of 25 species and in 108 cephalopods of 3 species purchased in Bayuquan region, Yingko city nearby the coast of the Bohai Sea from may to August 1992. A total of 7,327 larvae were collected from 156 fish of 19 species and 8 squids of one species. The 3rd-stage larvae of Anisakis simplex were collected from 121 fish (63.4%) of 15 species (N = 191) and from 8 squids (14.8%) of one species (N = 54), and they were total, 5,992 (81.8%). Out of remaining 1,335 larvae, 154 (2.1%) were classified as Thynnascaris type B from 23 fish of 4 species, 1,013 (13.8%) as Thynnascaris type C from 79 fish of 13 species. 164 (2.2%) as Hysterothylacium China type V from 20 fish of 4 species, 3 (0.04%) as Raphidascaris from 3 fish of 2 species and one was Pseudoterranova decipiens larva.
Animals↗
Calculation of the dose in the build-up region for high energy photon beam. Treatment planning when beam spoilers are employed.
Recoil electrons from a polystyrene sheet (beam spoiler) placed in a photon beam were used to modulate the dose in the build-up region. The effects of the beam spoiler on the dosimetry in simple phantoms were studied for both 6 MV and 15 MV photon beams at 100 cm source-to-surface distance (SSD) as a function of (1) the thickness of the spoiler, (2) the spoiler-to-surface distance (SD) and (3) the field size. A radiotherapy treatment planning (RTP) algorithm was used that employs pencil beam dose deposition kernels to evaluate the three-dimensional (3D) dose distribution due to photons and electrons in the build-up region. To determine the energy deposition kernels for the recoil electrons emanating from the spoiler, their energy spectrum needs to be known. Monte Carlo simulations were performed to derive the spectrum of electrons from the beam spoiler. The recoil electron spectrum was found to be nearly independent of field size, SD, and spoiler thickness. The pencil beam energy deposition kernel was calculated for the recoil electrons from the spoiler and inserted in the treatment planning algorithm. The dose calculation algorithm permits merging the photon and recoil electron dose components. In all cases the calculation agrees to within 3% of the measured results.
Algorithms↗
Sialidase in rabbit blood. Characterization of sialidase purified from rabbit erythrocyte membrane.
Sialidase activities of rabbit blood cells and serum were measured. The leucocyte particulate fraction showed the highest specific activity of sialidase towards mixed gangliosides and sialyllactose, and the cytosolic fraction showed for fetuin. Predominant sialidase activity in the blood was detected in erythrocyte particulate fraction when mixed gangliosides were used as substrate. The sialidase for ganglioside was solubilized from the erythrocyte ghosts by using Triton X-100. The solubilized sialidase was purified 1886-fold by sequential chromatographies on DEAE-cellulose, EAH-Sepharose 4B, Octyl-Sepharose CL-4B, Sephadex G-100, concanavalin-A--Sepharose, N-(p-aminophenyl)oxamic acid-agarose and Heparin-Sepharose CL-6B. The optimum pH of purified sialidase was 4.5 for ganglioside mixture, and this enzyme exhibited M(r) = 48,000 by gel filtration. When the purified sialidase was subjected to SDS/PAGE, a major sialidase-active protein band at M(r) = 54,000 and another fainter inactive protein band with M(r) = 115,000 were observed. The purified enzyme was active towards oligosaccharides, gangliosides, fetuin glycopeptide and 4-methylumbelliferyl alpha-D-N-acetylneuraminic acid except for glycoproteins tested. Fe2+, Fe3+ and dithiothreitol significantly inhibited the enzyme activity, while Triton X-100 activated the enzyme. Inside-out vesicles and unsealed ghosts of rabbit erythrocyte showed the sialidase activity for mixed gangliosides but not for resealed ghosts or intact erythrocytes. These results indicate that the active site of this sialidase is oriented mainly on the inside of the erythrocyte membrane and not on the outside. Treatment of rabbit erythrocyte unsealed ghosts with phosphatidylinositol-specific phospholipase C liberated no sialidase activity toward mixed gangliosides from the ghosts.
Animals↗
[Effect of glycyrrhetinic acid on DNA damage and unscheduled DNA synthesis induced by benzo (a) pyrene].
Glycyrrhetinic acid (GA) is an active component of Glycyrrhiza uralensis Fisch. In this study, the effects of glycyrrhetinic acid on DNA damage and unscheduled DNA synthesis induced by benzo (alpha) pyrene were studied. Mouse ear edema was visible obviously at the 6th h after topical application of single dose croton oil. A topical application of croton oil on the back of ICR mice for 5 h, induced elevation of ornithine decarboxylase (ODC) activity in epidermal. The administration of glycyrrhetinic acid (50-200 mg.kg-1.d-1) to animals for 3 days exhibited 20%-80% inhibition of epidermal ornithine decarboxylase activity in a dose-dependent manner. Benzo (alpha) pyrene obviously caused DNA damage and unscheduled DNA synthesis mediated by S9 fraction in Chinese hamster lung cell line. Glycyrrhetinic acid was found to protect the rapid DNA damage induced by benzo (alpha) pyrene. At concentration of 5 micrograms.ml-1, glycyrrhetinic acid exhibited 70% protection. At 20 micrograms.ml-1, this action was more potent and approached to 80%. The addition of hydroxyurea 10 mmol.L-1 suppressed DNA replicative synthesis to 84.04% and benzo (alpha) pyrene stimulated the DNA repair synthesis (6-fold). Glycyrrhetinic acid (20 micrograms.ml-1 and 50 micrograms.ml-1 significantly decreased the stimulation of DNA repair synthesis induced by benzo (alpha) pyrene. This suggests that glycyrrhetinic acid has effective anti-initiating and anti-promoting activities and could be used for cancer chemopreventive purpose.
Animals↗
Combining multileaf fields to modulate fluence distributions.
PURPOSE: A method for modulating beam fluence from a linear accelerator is discussed. The beam modulation is accomplished remotely using a multileaf collimator and does not require entering the treatment room. METHODS AND MATERIALS: The multileaf collimator is used to define a series of field shapes that are superimposed at a fixed gantry angle to produce any desired fluence pattern. A heuristic technique for deriving the field shapes and corresponding monitor unit settings is described. The technique has been tested on randomly generated fluence distributions and on distributions with a limited number of peaks and valleys. The second type of distribution more closely simulates fluence patterns obtained with dose optimization software. Estimates of the time required to use this approach to treat a four-field plan are given and compared to the technique of placing a physical compensator in each beam. RESULTS: It has been demonstrated that complex fluence patterns within a 15 x 15 cm2 field can be achieved with less than 20 fields. Estimates show that this technique is faster than entering the treatment room to change physical compensators. Some limitations of the method are discussed. CONCLUSION: Optimized distributions that conform the dose to irregularly shaped target volumes that wrap around critical structures are possible using superimposed multileaf fields. A method for defining the field shapes is presented.
Humans↗
Pattern and duration of the inhibitory effect of alcohol administered acutely on suckling-induced prolactin in lactating rats.
We have characterized the pattern and duration of the inhibitory effect of acute alcohol administration on suckling-induced prolactin (PRL) release in the lactating rat. On day 2 of lactation, litters were adjusted to eight pups. On day 6, dams were implanted with an atrial catheter and experiments were conducted on day 10 of lactation. Pups were removed from the dams at 0800 hr. An extension tube filled with heparinized saline was attached to the catheter at 1300 hr. At 1400 hr, a preinfusion (PRE 0) blood sample was removed and was followed by infusion of saline (control) or alcohol in saline (0.5, 1.0, 2.0, 2.5 g/kg body weight doses) solutions. Following the removal of a postinfusion (POST 0) blood sample, pups were returned to the mother. Subsequent blood samples were obtained 10, 30, 60, 120, and 180 min after initiation of suckling. In separate groups, the effects of alcohol on basal PRL were studied by collecting blood samples PRE 0, POST 0 and 20, 40, 60, 80, 100, and 120 min following infusion of saline or alcohol in saline to lactating rats also separated from their pups for 6 hr. Alcohol infusion did not alter basal PRL. However, suckling-induced PRL was inhibited at 10, 30, 60, and 120 min of suckling by alcohol administered at doses greater than or equal to 1.0 g/kg body weight. After 180 min of suckling, plasma PRL levels were comparable among groups. The suckling latency for the 2.5 g/kg body weight alcohol group was greater than for other groups, but the quantities of milk consumed during the 3-hr suckling period were comparable.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
[Influence of the active compounds isolated from pilose antler on syntheses of protein and RNA in mouse liver].
The polyamines of pilose antler (PASPA) consist of putrescine (PU, 70.9%), spermidine (SPD, 26.3%) and spermine (SP, 2.8%). The incorporations of [3H] leucine into protein and [3H] uridine into RNA in mouse liver tissue were increased when PASPA was given orally to mice at the dose of 30 mg/kg for 4 successive days. The incorporations of [3H] leucine into liver protein and [3H] uridine into the cytosolic and nuclear RNA were also increased by treatment with PU (21 mg/kg). In addition, the RNA polymerase activity in the solubilized liver nuclear fraction of PU (21 mg/kg)-treated mice was increased. SPD only promoted the synthesis of protein in mouse liver tissue at the dose of 8 mg/kg. However, SP showed no effect on the synthesis of protein and RNA polymerase activity under the used dose (1 mg/kg). The results suggest that PASPA is the main active substance responsible for the promotion of the synthesis of protein and RNA in mouse liver.
Animals↗
[Effect of polyamines isolated from pilose antler (PASPA) on RNA polymerase activities in mouse liver].
The incorporations of [3H] leucine into protein and [3H] uridine into RNA in mouse liver were increased when PASPA was given to mice at a dose of 30 mg/kg for 4 successive days. The RNA polymerase activity, especially the RNA polymerase II activity in the solubilized liver nuclear fraction of PASPA-treated mice was also increased. In vitro experiment demonstrated that PASPA increased the RNA polymerase activity significantly in mouse liver nuclei at a concentration of 1 microgram/ml. These results suggest that the enhancement of RNA polymerase activities, particularly RNA polymerase II activity, induced by PASPA treatment is responsible for the increase in synthesis of protein and RNA in mouse liver tissue.
Animals↗
Twist drill puncture and fragment aspiration with simple stereotaxic guidance in treatment of patients with hypertensive intracerebral hematomas.
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Adult↗
[Inhibitory effect of hypoxanthine on monoamine oxidase activity].
Hypoxanthine was demonstrated to have a dose-dependent inhibitory effect on type B monoamine oxidase (MAO-B) activity in liver and brain tissues, and slight inhibitory effect on type A MAO(MAO-A) activity when given orally to mice at doses of 25-500 mg/kg. When mice were given orally hypoxanthine 500 mg/kg, MAO-A and -B activities were all inhibited significantly 16 hours after administration, but the inhibitory action on MAO-A was weaker. Subcutaneous injection of the agent also produced obvious inhibition of MAO activity in the liver but no significant influence on MAO activity in brain tissue was observed. In vitro experiment showed that the action of hypoxanthine on MAO-B was competitive inhibition and that on MAO-A was competitive mixed with noncompetitive inhibition.
Animals↗