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Biomedical subjects

F Lin

Publications and source records attributed to F Lin.

At least 73 records · Page 4Linked to original sources

Relationship between bone mineral density and polymorphism of the estrogen receptor gene in healthy postmenopausal women in China.

OBJECTIVE: To investigate the possible relationship between bone mineral density and polymorphism of the estrogen receptor (ER) gene in Shanghai healthy postmenopausal women. METHODS: 250 unrelated healthy postmenopausal women were selected for bone mineral density (BMD) determination by Dual energy X-ray absorptiometry (DEXA) and polymorphism of estrogen receptor gene analyses by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: Pvu II polymorphisms of ER gene was associated with low Troch BMD (P = 0.0153) while there was no significant relationship between Xba I polymorphism of ER gene and BMD at any of skeletal sites included in the present study, and the combination of Pvu II and Xba I polymorphisms of ER gene was significantly associated with both low Lumbar 2-4 (P = 0.0369) and Troch (P = 0.0384) BMD. Multiple stepwise regression analysis also indicated that two combined polymorphisms were correlated significantly with Lumbar 2-4 BMD (P = 0.0254) while this correlation was not revealed at any other skeletal sites. CONCLUSION: There is significant relationship between the polymorphism of ER gene and both Lumbar 2-4 BMD and Troch BMD. It is significant to explore the pathogenesis of osteoporosis and to prevent the development of osteoprosis by use of molecular genetics.

Absorptiometry, Photon↗

Hemin-enhanced resistance of human leukemia cells to oxidative killing: antisense determination of ferritin involvement.

Human HL-60 cells exhibited a strong hyperresistance to the lethal effects of photodynamic activity (singlet oxygen) or glucose oxidase activity (hydrogen peroxide) 16-20 h after being exposed to hemin (ferriprotoporphyrin IX). Hyperresistance was accompanied by the overproduction of immunodetectable ferritin, predominantly the heavy (H) subunit, which exhibits ferroxidase activity. Cells that had been enriched in apoferritin via pinocytotic uptake showed similar hyperresistance to both types of oxidative challenge. On the other hand, preincubating cells with hemin in the presence of a phosphorothioate-linked antisense oligodeoxynucleotide against H-ferritin mRNA resulted in a strong diminution in both hyperresistance and H-ferritin induction. No effects were seen when a scrambled order oligodeoxynucleotide of the same base composition was used, confirming that the antisense oligomer had specifically inhibited H-ferritin translation. These results indicate that induced ferritin played a crucial role in the observed cytological responses. Enhanced oxidant resistance is attributed to the ability of this ferritin to rapidly sequester and incapacitate redox-active iron.

Apoferritins↗

Androgenesis and homozygous gynogenesis in muskellunge (Esox masquinongy): evaluation using flow cytometry.

The purpose of this work was to study the effects of ultraviolet (UV) irradiation on denucleation of eggs and investigate the heat-shock conditions for diploidization for induction of androgenesis in muskellunge, Esox masquinongy. Several egg incubation media, including saline, Ringer's solution, and Ringer's solution supplemented with bovine serum albumin (BSA), were found suitable to maintain the egg fertility as high as in muskellunge ovarian fluid. The optimal doses of UV radiation were 660-1320 J/m2, at which 100% haploid larvae were produced at a hatching rate of 22.5 +/- 2.8%. UV irradiation at low doses (165-330 J/m2) generated abnormal larvae, which were morphologically identical to haploids. Using a flow cytometry method, it was found that cellular DNA content of these larvae was close to that of diploids but significantly lower in value and had a wider distribution (expressed as coefficient of variation) than that of control fish. This suggested that a low dose of UV irradiation might cause gene mutations, alteration of chromosomal conformation and fragmentation, but did not prevent maternal DNA from participating in mitotic division. Interference of maternal DNA residues could be another reason for the poor viability of androgenetic fish. A high dose of UV radiation (1980 J/m2) caused development of severely deformed embryos, indicating that UV radiation also damaged molecules in the eggs other than the denucleation. Our results suggest that classic color and allozyme markers might not be sufficient to prove a complete androgenesis. In order to optimize time and duration of shock for induced diploidization, we investigated the heat-shock conditions for inhibiting the first mitotic cleavage through induction of homozygous gynogenesis. We found that heat-shock treatment at 31 degrees C for 9 min starting at 1.4 tau 0 (a dimensionless factor describing progress in embryo development) after fertilization produced the highest percentage of diploids at hatching.

Animals↗

Radiographically ossified ganglion cyst of finger in a swimmer.

Ganglion cysts are fibrous-walled cystic lesions closely associated with joint or tendon sheaths and contain gelatinous mucinous fluid. The radiographic appearance is usually normal. Calcification or ossification in these cysts is extremely unusual. We report on an unusual appearing ganglion cyst of the little finger in a swimmer with ossification resembling myositis ossificans.

Diagnosis, Differential↗

The persistence of swine vesicular disease virus infection in pigs.

Two groups of pigs were infected with a recent Italian isolate of swine vesicular disease virus (SVDV). Blood, nasal swabs and faeces were collected for up to 6 months after exposure to infection and animals were killed at regular intervals to obtain tissues post-mortem. These samples were examined for virus by conventional means and for viral RNA (vRNA) by reverse transcription-nested polymerase chain reaction (RT-nPCR). Virus was identified intermittently from both clinically and subclinically infected animals in nasal swabs, faeces and tonsillar tissue by either virus isolation or RT-nPCR up to 63 days post infection (dpi). Between 63 and 119 dpi virus was not detected in the secretions, excretions or tissues of any pigs. Following mixing of the two groups of animals at 119 dpi, SVDV was again identified in faeces for up to 7 days suggesting that the stress of mixing reactivated the excretion of virus in pigs from which the agent could no longer be identified. Minor antigenic changes were identified between the parental virus and isolates recovered late in the course of infection. Altered antigenicity corresponded with deduced amino acid substitutions identified from differences in nucleotide sequence between early and late isolates. This investigation demonstrates that SVDV and vRNA can be present in pigs for considerably longer after exposure to infection than has previously been recognized and provides preliminary evidence for a carrier state in swine vesicular disease.

Amino Acid Substitution↗

Adoptive immunotherapy for relapse of chronic myeloid leukemia after allogeneic bone marrow transplant: equal efficacy of lymphocytes from sibling and matched unrelated donors.

Lymphocyte transfusion from the marrow donor (DLT) is well established as an effective therapy for relapse of CML post allogeneic BMT. Reports thus far have been mostly limited to patients who received DLT from a matched sibling donor. We compared the efficacy and toxicity of DLT in 30 patients who were treated with cells from their HLA-identical sibling (n = 18) or from their phenotypically HLA-matched unrelated marrow donor (n = 12). The overall probability of obtaining a cytogenetic remission was 69% (95%CI: 51-83%) and was not significantly different between the two groups. The disease stage at the time of DLT was the only factor associated with cytogenetic remission by multivariate analysis; patients treated in cytogenetic or molecular relapse (n = 11) were seven times more likely (RR = 7.4, 95%CI: 2.4-22.4, P = 0.0005) to respond compared to patients treated for hematologic relapse (n = 19). There was a trend towards more acute GVHD II-IV in the unrelated donor group (58 vs 39%, P = 0.09), but the probability of developing extensive chronic GVHD was not significantly different (56 vs 39%, P = 0.4). We conclude that transfusion of donor cells from HLA-matched volunteer donors does not appreciably increase the risk of GVHD compared with transfusion of cells from HLA-identical siblings in patients with CML who relapse following allogeneic BMT. Conversely, there is no evidence for an increased graft-versus-leukemia effect after DLT from volunteer donors.

Adult↗

Far UV photolysis of uracil and cytosine in phosphate solution.

The photolysis of nucleobases, nucleosides and nucleotides (NA) was enhanced by phosphate under irradiation of medium pressure mercury lamp (MPML). Uracil and Cytosine in phosphate solution were selected to study the mechanism of phosphate effect. Photoproducts were produced in the irradiated uracil and cytosine of phosphate solution which have been isolated by anion exchange resin. Ultraviolet irradiation (190-220nm) of uracil in 0.05 mol/dm3 phosphate buffered solution at pH 8-9 leads to the production of a novel compound C4H5N2O6P which has been identified by UV, 1H-NMR spectroscopy and LC/MS/MS. The formation mechanism of the photoproduct and the kinetics were studied.

Amino Acids↗

[Classification and related operation procedures of pelvic tumor treated with partial pelvic resection].

OBJECTIVE: To establish a classification system of pelvic tumor in terms of the location of lesion in order to project surgical treatment with less extent of damage to the affected limb function. METHOD: Thirty-six cases were treated by partial pelvic resection with limb salvage, and 20 were followed up. The lesions were divided into four groups on the basis of sacral-iliac and/or hip joint involved. In group I, lesion was noted in the ischium or pubis only;in group II the lesion was located at sacral-iliac joint or hip joint. The patients with acetabulum involved were classified into group III both sacral-iliac joint and hip joint were destroyed by tumor in group IV. Surgical treatment was designed as resection, resection with sacral-iliac joint reconstruction, resection without hip joint reconstruction according to the classification of the lesion. The patients in group IV were treated with traditional hemi-pelvic amputation. RESULT: Patients were followed up from 4 months to 14 years (average 51.7 months). The limb function was preserved to some extent, and pelvic stability was satisfactory. Walking gait was not chlanged significant in group I and II. But resection without hip joint reconstruction in group III resulted in limb discrepancy (4.0 - 8.0 cm) and walking lamely. The patients needed custom-made shoes with heightened heel, but some of them walked with crutch when the discrepancy exceeded 8 cm. There were no nerve and vessel injury in the long run. The strength of muscles around hip joint was III - IV grade after postoperative exercise, and the capability of walking and viability recovered 3 months after the operation. CONCLUSION: Different surgical treatments should be selected according to the site of pelvic tumor. The classification system depending on the lesion's location in pelvic tumor is useful in selecting operative procedure to retain limb function to great extent.

Adolescent↗

[Seeds collecting of Ephedra sinica and treatment before sown].

Seeds collecting of Ephedra sinica and treatment before sown were studied. These results indicated that time collecting seeds and treating such as threshing, washing, drying, storing at low temperature and immersing seeds with chemical were available measures. Which could supply a reference for the culture of E. sinica.

Desiccation↗

Pharmacological characterization of the human ionotropic glutamate receptor subtype GluR3 stably expressed in mammalian cells.

We have cloned the human ionotropic alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor GluR3 flip splice variant (hGluR3i) and developed a stable cell line expressing this receptor in HEK293 cells. Electrophysiological recordings demonstrated that glutamate-evoked currents desensitize rapidly, with a mean desensitization time constant of 5.4 ms. Robust glutamate-evoked increases in intracellular Ca++ ([Ca++]i) were observed in the presence of cyclothiazide, which attenuated receptor desensitization. [Ca++]i measurements were used to perform a detailed pharmacological characterization of hGluR3i with reference agonists and antagonists. The results of these studies showed that kainate and domoate were not fully efficacious agonists relative to glutamate. The binding affinities of agonists and competitive antagonists were determined in a [3H]AMPA competition binding assay. There was a good correlation between the functional data and the binding affinities obtained for competitive antagonists. However, the binding affinities of the agonists did not correlate with their functional EC50 values from [Ca++]i data, possibly because the binding assay predominantly measures the desensitized high-affinity state of the receptor. [3H]AMPA binding also was performed on membranes prepared from rat forebrain, and comparison of the data from HEK293 cells expressing hGluR3i and rat forebrain suggest that nearly all of the reference compounds show similar binding activities between the two membrane preparations, with the exception of fluoro-willardiine, kainate and 6-nitro-7-sulfamoylbenzo(f)quinoxaline-2-3-dione (NBQX). These data suggest that cells stably expressing recombinant hGluR3i represent pharmacologically valid experimental systems to study human AMPA receptors.

Animals↗

Delayed hyperresistance of endothelial cells to photodynamic inactivation after contact with hemin.

Hemin (ferriprotoporphyrin IX), the oxidized prosthetic group of hemoglobin, is a potential source of prooxidant iron in heavily vascularized tumors. We have evaluated hemin's effects on photodynamic inactivation of bovine artery endothelial cells, using a partially purified oligomeric fraction of hematoporphyrin derivative (HPD-A) as the sensitizing agent. Confluent cells in 5% serum/RPMI medium showed a progressive loss of thiazolyl blue (MTT)-detectable viability when irradiated with broadband visible light in the presence of HPD-A. Cells pretreated with desferrioxamine (DFO) were substantially less sensitive to photokilling, implying that non-heme iron plays a role in cytotoxic activity. Hemin (10-20 microM) had remarkably different effects on photokilling, depending on the time interval between adding it to cells and exposing them to photodynamic action. For example, cells were more sensitive when photostressed immediately after 1 h hemin treatment and washing but much more resistant when photostressed 23 h later. Similar responses were observed when cells were challenged with glucose oxidase. Immunoblot analysis following hemin treatment revealed a progressive induction of the heavy (H) subunit of ferritin that paralleled the development of hyperresistance. After incubation with saturating levels of the synthetic iron donor [55Fe]ferric-8-hydroxyquinoline, hemin-stimulated cells contained about four times more immunoprecipitable ferritin 55Fe than controls. This is consistent with the notion that sequestration of toxic iron as a result of induction of H-chain-enriched ferritin is a key factor in hyperresistance. Inflammatory injury in tumor vasculatures could expose endothelial and neoplastic cells to chronic hemoglobin-derived iron. Consequent upregulation of ferritin could impact negatively on the efficacy of photodynamic therapy and other oxidant-based cancer therapies.

Animals↗

Expression of nuclear lamins in human tissues and cancer cell lines and transcription from the promoters of the lamin A/C and B1 genes.

We have examined the expression of lamins A, B1, and C in human tissues and cancer cell lines and the function of the lamin A/C and B1 gene promoters in transfected cells. Northern analysis and immunoblotting demonstrated that lamin A/C mRNA and protein were not detectable in some human cell lines whereas lamin B1 was always present. Sequencing of approximately 2.6 kb of the lamin A/C and 1.6 kb of the lamin B1 genes 5' to the translation initiation sites showed that they did not contain typical TATA boxes near the transcription start sites. The lamin B1 and A/C proximal promoter regions were transcribed in transfected HeLa, Raji, and NT2/D1 cell lines even if the cells did not contain detectable endogenous lamin A/C mRNA or protein. These results show that, similar to most cytoplasmic intermediate filament genes, transcriptional regulatory elements in the promoters of the human nuclear lamin A/C and B1 genes do not control their cell type-specific expression in culture lines.

Base Sequence↗

Elevated ferritin production, iron containment, and oxidant resistance in hemin-treated leukemia cells.

Hemin (ferriprotoporphyrin IX), the oxidized prosthetic group of hemoglobin, is a source of potentially cytotoxic iron, but in chronic low doses can induce cytoprotection against iron-stimulated oxidative stress. The latter property of hemin has been examined, using murine L1210 cells and three different oxidant generating systems: (i) glucose/glucose oxidase, (ii) near-ultraviolet irradiation, and (iii) dye-mediated photodynamic action. Cells treated with the lipophilic iron donor ferric-8-hydroxyquinoline, Fe(HQ)2 (1 microM, 30 min) were found to be more sensitive to oxidative killing than nontreated controls. However, cells challenged after long-term (20-24 h) exposure to hemin (10 microM) were substantially more resistant than controls and were sensitized far less by Fe(HQ)2. Immunoblot analyses of 24-h hemin-treated cells indicated that the ferritin heavy (H) subunit was elevated 12- to 15-fold, whereas the light (L) subunit was essentially unchanged. Experiments carried out with 55Fe(HQ)2 showed that iron uptake capacity of cells was greatly enhanced after hemin treatment. More specifically, hemin-stimulated cells were found to contain approximately 9 times more immunoprecipitable ferritin iron after incubation with saturating levels (4-5 microM) of 55Fe(HQ)2 and approximately 3 times more iron per ferritin molecule compared with nonstimulated controls. The nonferritin iron content of the latter was estimated to be approximately 40 times greater than that of the former following low-level (0.5 microM) 55Fe(HQ)2 treatment. These results are consistent with the idea that induced ferritin, enriched in H-chain, sequesters redox active iron rapidly and copiously, thereby enhancing cellular resistance to oxidants.

Animals↗

Detection of swine vesicular disease virus RNA by reverse transcription-polymerase chain reaction.

Two polymerase chain reaction (PCR) assays are described for the detection of swine vesicular disease virus (SVDV) RNA, a reverse transcription PCR (RT-PCR) and a reverse transcription nested PCR (RT-nPCR). Both the RT-PCR and RT-nPCR were able to detect representative members of each of seven phylogenetically distinct groups of SVDV and gave negative results with a range of porcine enteroviruses and of viruses responsible for vesicular conditions in pigs. When combined with a commercial kit for rapid RNA extraction, the RT-PCR was useful for the detection of SVDV in samples of epithelium and faeces from animals with clinical SVD. The addition of a second amplification step to create a nested PCR (RT-nPCR) increased the sensitivity of the technique for the detection of viral RNA (vRNA) in SVDV infected tissue culture fluid by a factor of approximately 1,000, for 100 TCID50 for the RT-PCR to 0.1 TCID50 for RT-nPCR. When combined with a more elaborate extraction procedure for RNA, the RT-nPCR was considerably more sensitive than virus isolation in tissue culture for detecting SVDV in nasal swabs, tissues, and faeces collected from pigs between 7 days and 176 days after infection with a recent European isolate of SVDV. However, stringent conditions are necessary for carrying out the RT-nPCR to minimise the possibility of contamination.

Animals↗

Lipid peroxidation in photodynamically stressed mammalian cells: use of cholesterol hydroperoxides as mechanistic reporters.

Photodynamic action of merocyanine 540, an antileukemic sensitizing dye, on murine L1210 cells results in the formation of lipid hydroperoxides and loss of cell viability. High-performance liquid chromatography with mercury cathode electrochemical detection was used for determining lipid oxidation products, including the following cholesterol-derived hydroperoxides: 5 alpha-OOH, 6 alpha-OOH, 6 beta-OOH, and unresolved 7 alpha, 7 beta-OOH. Among these species, 5 alpha-, 6 alpha-, and 6 beta-OOH (singlet oxygen adducts) were predominant in the early stages of photooxidation, whereas 7 alpha- and 7 beta-OOH (products of free radical reactions) became so after prolonged irradiation or during dark incubation after exposure to a light dose. These mechanistic changes were studied in a unique way by monitoring shifts in the peroxide ratio, i.e., 7-OOH/5 alpha-OOH, or 7-OOH/6-OOH. When cells (10(7)/ml) were exposed to a visible light fluence of 0.6 J/cm2 in the presence of 10 microM merocyanine 540, 7-OOH/5 alpha-OOH increased by approximately 100% after 2 h of dark incubation at 37 degrees C. The increase was much larger (approximately 250%) when cells were photooxidized after treatment with 1 microM ferric-8-hydroxyquinoline, a lipophilic iron donor, whereas no increase was observed when cells were pretreated with 100 microM desferrioxamine, an avid iron chelator/redox inhibitor. Correspondingly, postirradiation formation of thiobarbituric acid-reactive material was markedly enhanced by ferric-8-hydroxyquinoline and suppressed by desferrioxamine, as was the extent of cell killing. When added to cells after a light dose, chain-breaking antioxidants such as butylated hydroxytoluene and alpha-tocopherol strongly protected against cell killing and slowed the increase in 7-OOH/5 alpha-OOH ratio. It is apparent from these results that (1) the 7-OOH/5 alpha-OOH or 7-OOH/6-OOH ratio can be used as a highly sensitive index of singlet oxygen vs. free radical dominance in photodynamically stressed cells; and (2) that postirradiation chain peroxidation plays an important role in photodynamically initiated cell killing.

Animals↗

The rate and kinetics of molecular response to donor leucocyte transfusions in chronic myeloid leukaemia patients treated for relapse after allogeneic bone marrow transplantation.

We have assessed the molecular response of 30 consecutive patients with chronic myeloid leukaemia (CML) treated for relapse after allogeneic bone marrow transplantation (BMT) by donor leucocyte transfusions (DLT). Response was evaluated by qualitative nested and quantitative competitive RT-PCR for BCR-ABL mRNA at various time intervals before and after DLT. The probability of attaining molecular remission at 2 years was 61% (95% CI 42-78%). Disease state at the time of DLT was significantly associated with response: molecular remission was achieved for 9/10 (90%) patients treated early (cytogenetic or molecular relapse) compared to only 8/20 (40%) patients treated late (haematological relapse; P = 0.009). The Kaplan-Meier estimates of molecular remission at 2 years post DLT for patients treated in early or late relapse were 86.6% and 47.3% respectively (P = 0.004). The median time interval from DLT to molecular remission was 11.0 months (range 2.5-32). Molecular remissions were durable for most (15/17) patients (median follow-up 21.2 months; range 0-55). Two patterns of molecular response were found: a very rapid decline after an initial lag phase or a more gradual decline over a period of several months. We conclude that molecular monitoring is a sensitive indicator of response to DLT; different kinetics of molecular response may reflect disease heterogeneity or differences in the mode of action of DLT.

Bone Marrow Transplantation↗