[Studies on the constituents of Erycibe elliptilimba].
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Biomedical subjects
Publications and source records attributed to Y Lu.
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When K562 cells were made resistant to cisplatin their neutral amino acid transport systems changed. K562 cells cloned in cisplatin developed a 6.7-fold resistance to the drug. The generation times for K562 cells sensitive (S) and resistant (DDP) to cisplatin were similar. The initial uptake and rapid efflux of cisplatin were similar in both cell lines. However, they differed in their sodium dependent neutral amino acid transport properties. In K562S cells the sodium-dependent uptake for methylaminoisobutyric acid was significantly inhibited by threonine (Ki = 10.3 mM), but in K562DDP the uptake of neutral amino acids was significantly lower, and methylaminoisobutyric acid uptake was minimally inhibited by threonine. When K562DDP cells were grown in the absence of cisplatin for 8 weeks, their amino acid transport properties reverted to those of K562S cells. In K562 cells, changes in plasma membrane essential amino acid transport systems were concomitant to both the development of resistance and the redevelopment of sensitivity to cisplatin. Therefore, human malignant cell sensitivity and resistance to cisplatin may be related to membrane nutrient transport systems.
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DNA restriction fragments ranging from 79 to 789 base pairs in length have been characterized by transient electric birefringence (TEB) measurements at various temperatures between 4 and 43 degrees C. The DNA fragments do not contain runs of four or more adenine residues in a row and migrate with normal electrophoretic mobilities in polyacrylamide gels, indicating that they are not intrinsically curved or bent. The low ionic strength buffers used for the measurements contained 1 mM Tris Cl, pH 8.0, EDTA, and variable concentrations of Na(+) or Mg(2+) ions. The rotational relaxation times were obtained by fitting the TEB field-free decay signals with a nonlinear least-squared fitting program; the decay of the birefringence was monoexponential for fragments < or = 241 base pair (bp) in length and multiexponential for larger fragments. The terminal relaxation times, characteristic of the end-over-end rotation of the DNA molecules, were then used to determine the persistence length (p) and hydrodynamic radius (r) of DNA as a function of temperature and ionic strength, using several different hydrodynamic models. The specific values obtained for p and r are model dependent. The wormlike chain model of P. J. Hagerman and B. H. Zimm (Biopolymers 1981, Vol. 20, pp. 1481-1502) combined with the revised Broersma equation (J. Newman et al., Journal of Mol Biol 1997, Vol. 116, pp. 593-606) appears to be the most suitable for describing the flexibility of DNA in low ionic strength solutions. The values of p and r obtained from the global least squares fitting of this equation are independent of DNA length, and the deviations of the individual values from the average are reasonably small. The consensus r value calculated for DNA in various low ionic strength solutions containing 1 mM Tris buffer is 14.7 +/- 0.4 A at 20 degrees C. The consensus p values decrease from 814 approximately 564 A in solutions containing 1 mM Tris buffer plus 0.2-1 mM NaCl and decrease still further to 440 A in solutions containing 0.2 mM Mg(2+) ions. The persistence length exhibits a shallow maximum at 20 degrees C and decreases slowly upon either increasing or decreasing the temperature, regardless of the model used to fit the data. By contrast, the consensus values of the hydrodynamic radius are independent of temperature. The calculated persistence lengths and hydrodynamic radii are compared with other data in the literature.
A transforming gene was cloned from a focus of rat fibroblast cell line Rat 1 transfected with DNA of a gastrocarcinoma cell line. The transforming gene is the activated form of oncogene c-Ha-ras, and the lesion was identified as a single nucleotide substitution at codon 12 by sequencing. We further used polymerase chain reaction (PCR)-restriction analysis to detect the mutations at codon 12 of c-Ha-ras. By using this method, 11 cases out of 24 solid tumors and 3 cell lines of gastric cancer were shown to contain mutations, and there was a strong correlation between mutations and metastasis and survival of the patients. The role of the activated c-Ha-ras in tumorigenesis of stomach cancer was further proved through the assay in which the malignant phenotypes of the c-Ha-ras-transformed cells were partially inhibited by blocking the c-Ha-ras expression with antisense oligonucleotides or exogenous plasmid expressing antisense RNA.
The expression of p185erbB2 and p21ras was determined immunohistochemically in normal mucosa, intestinal metaplasia, dysplasia and in intestinal and diffuse type carcinoma of the stomach. The positive rates of p185erbB2 and p21ras in intestinal type were significantly higher than those of diffuse-type carcinoma. The results indicate that there are differences not only in clinical and biologic features, but also in molecular abnormalities between the two types of stomach cancer. Positive staining of cell membrane for p185erbB2 was observed specifically in intestinal type cancer, but not in the other lesions. Positive expression of p185erbB2 on the cell membrane appears to be a useful marker in identification of malignant change of gastric mucosa, as well as a late event of gastrocarcinogenesis. The results of in situ hybridization analysis in cancer tissues were consistent with those obtained by immunohistochemistry and demonstrated amplification of erbB2 gene at the mRNA level. No significant difference of p21ras expression was found among intestinal metaplasia, dysplasia, and intestinal type carcinoma; therefore, it might not be a specific marker of malignancy in gastric mucosa. Expression of p21ras may be an early event in the development of lesions predisposing to carcinoma.
In an attempt to prevent the sequelae of conventional gastrectomy, such as rapid gastric emptying, dumping syndrome, intestinal content reflux, indigestion, and poor absorption, we have devised the pylorus and pyloric vagus preserving gastrectomy (PPVPG). Experimenting on 48 dogs, we found theoretic grounds for using our design and obtained the desired effects--retaining the merits of conventional subtotal resection of the stomach, with acidity reduction, while avoiding the above-mentioned complications.
Thirteen cell lines were established and characterized from brain, kidney, lung, spleen, heart, liver, gall bladder, urinary bladder, pancreas, testis, skin, and periorbital and tumor tissues of an immature male green turtle (Chelonia mydas) with fibropapillomas. Cell lines were optimally maintained at 30 degrees C in RPMI 1640 medium supplemented with 10% fetal bovine serum. Propagation of the turtle cell lines was serum dependent, and plating efficiencies ranged from 13 to 37%. The cell lines, which have been subcultivated more than 20 times, had a doubling time of approximately 30 to 36 h. When tested for their sensitivity to several fish viruses, most of the cell lines were susceptible to a rhabdovirus, spring viremia carp virus, but refractory to channel catfish virus (a herpesvirus), infectious pancreatic necrosis virus (a birnavirus), and two other fish rhabdoviruses, infectious hematopoietic necrosis virus and viral hemorrhagic septicemia virus. During in vitro subcultivation, tumor-like cell aggregates appeared in cell lines derived from lungs, testis, and periorbital and tumor tissues, and small, naked intranuclear virus particles were detected by thin-section electron microscopy. These cell lines are currently being used in attempts to isolate the putative etiologic virus of green turtle fibropapilloma.
High resolution ultrasound imaging of the mouse placenta during development revealed highly echogenic foci localized near the materno-placental interface in early gestation and, near term, in the placental labyrinth (the exchange region of the placenta). Echogenic foci and calcium deposits identified in histological sections using Alizarin red staining showed similar localization and changes with gestation. Calcium deposits caused the echogenic foci because incubating uteri in a decalcifying solution eliminated both the deposits and echogenic foci. Transmission electron microscopy, X-ray microanalysis, and electron diffraction were used to show that deposits were calcium hydroxyapatite crystals. Calcium deposits were extensive and densely packed at days 7.5-9.5 of gestation at the border between the maternal decidua and the fetal trophoblast giant cells of ectoplacental cone. After the formation of the chorio-allantoic placenta (approximately day 10.5), calcification deposits appeared larger and more rarefied but were still localized at the border between the maternal decidua and the fetal trophoblast giant cells of the placenta. Calcification deposits were not observed in the labyrinthine region of the mouse placenta until > or = day 15.5 (day 18.5 is full term). We conclude that deposits of calcium hydroxyapatite crystals in the mouse placenta are detectable by high resolution ultrasound imaging. These deposits provide an ultrasound detectable marker of the maternal-placental interface that is particularly prominent during the establishment of the chorio-allantoic placenta between days 7.5 and 9.5 of gestation.
This study focused on whether bone marrow would differentiate tolerogenic dendritic cells (DCs) by comparing interleukin-3 (IL-3) DCs (induced by IL-3+IL-4) with GM-CSF DCs (induced by GM-CSF+IL-4, generating myeloid DCs). Mouse bone marrow progenitors were induced in vitro to differentiate into DCs by exposure to IL-3. The morphology and yields of IL-3 DCs and GM-CSF DCs were compared by scanning electron microscopy and laser scanning confocal microscopy. Immunohistochemistry and flow cytometer (FCM) were used to identify immune phenotypes. The results showed that IL-3 DCs and GM-CSF DCs both expressed the specific marker DEC-205 with about 80% and 70% purity, respectively. IL-3 DCs had similar morphology as plasma cells, but a different immune phenotype, namely, CD80(-), CD86(-), CD14(-), and MHC class II(+). IL-3 DCs remained in the costimulatory molecule-deficient state when TNF-alpha was supplemented into the culture. We therefore propose that IL-3 induces a high purity and yield of DCs from bone marrow. Since IL-3 DCs possess a tolerogenic phenotype (MHC class II(+), B7(-)), they may induce alloantigen-specific hyporesponsiveness in vitro.
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