Biomedical subjects

L Meng

Publications and source records attributed to L Meng.

At least 37 records · Page 2Linked to original sources

PubMed2000

A case-control study of mammographic densities in Hawaii.

Epidemiologic evidence suggests that mammographic densities are markers of breast cancer risk. This project investigated the relation between breast cancer and densities in women of Chinese, Japanese, Filipino, Native Hawaiian, and Caucasian ancestry. Mammograms from breast cancer cases and from healthy controls were compared using a computer-assisted method of mammographic density assessment. From 1991 to 1997, 935 cases of breast cancer were diagnosed at Kaiser Permanente in Hawaii; for 647 (69%) subjects, a control woman matched by ethnicity, year of mammogram, and age was identified. Conditional multiple logistic regression was applied to estimate the relative risk of developing breast cancer. Breast cancer risk was associated with percent densities and with the size of the dense areas. Women in the category with the most densities experienced a twofold risk of developing breast cancer as compared to women with the least densities. Adjustment for risk factors reduced the strength of the association. Odds ratios were of similar magnitude in Asian women as in Caucasian/Native Hawaiian women, but they were not statistically significant. The results of this study indicate that the associations of breast cancer risk with the magnitude of the dense areas and with the percent densities are of similar strength in women of different ethnicities although density levels vary by ethnicity.

Adult↗

PubMed2000

[Ecological effect of CdCl2 on plasmid and role of plasmid in Cd-tolerance of its host].

After treating the plasmid of Escherichia coli HB101 by CdCl2 in vitro or in vivo, the effect of Cd on the structure of plasmid pWH58 DNA was studied through argarose gel electrophesis and restriction fragment length polymorphism analysis. By comparing the growth of E. coli with and without plasmid cultured in the medium with Amp LB and non-anti LB under different concentrations of CdCl2, the effect of Cd on plasmid E. coli of in vivo and the role of plasmid in Cd-tolerance of its host were studied. Cd treatments of E. coli in vitro or in vivo didn't cause an obvious change of the structure of plasmid pWH58 DNA. Plasmid pWh58 could replicate for regeneration and gene express under Cd stress. Plasmid pWH58 of E. coli in vivo lowered Cd-tolerance of its host form 75 mg.L-1 to 50 mg.L-1. Cd-tolerance of E. coli ascended markedly after Cd taming, but had a trend of reversing to original level after restoration, indicating that Cd-tolerance of E. coli could be tamed, but the tamed Cd-tolerance still had no genetic basis.

Cadmium Chloride↗

PubMed2000

[A histopathologic and immunohistochemical study of prognostic factors in esophageal squamous cell carcinoma].

OBJECTIVE: To investigate the prognostic value of various histopathological features and expression of immuno-phenotypes in esophageal squamous cell carcinoma (ESCC). METHODS: The tumor samples from 97 primary ESCC patients with potentially curative resection were evaluated for the following parameters: tumor size, grade of differentiation, pattern of invasion, depth of invasion, degree of lymphocyte infiltration, lymph node metastasis, TNM stage and p53, EGFR, c-erbB-2, nm23, cathepsin D protein expression. The prognostic impact of these parameters were analyzed by univariate and multivariate survival analyses. RESULTS: In the univariate Kaplan Meier analysis, poor differentiation, infiltrating growth pattern, invasion reaching adventitia, absent lymphocytic infiltration, lymph node metastasis, high staging, EGFR positive expression and nm23 negative expression were all associated with lower survival rate (Log-rank test, P < 0.05, respectively). However, the multivariate Cox analysis demonstrated that only pattern of invasion (RR = 2.000), degree of lymphocyte infiltration (RR = 0.509), TNM stage (RR = 1.733), EGFR expression (RR = 1.812) and c-erbB-2 expression (RR = 2.364) were significant prognostic factors (P < 0.05, respectively). CONCLUSION: For ESCC, the pattern of invasion, degree of lymphocyte infiltration, EGFR and c-erbB-2 protein expression had independent prognostic values as well as the TNM stage. Comprehensive analysis of these parameters can be helpful to identify biological behavior of ESCC and predict more accurately the prognosis of patients with ESCC.

Adult↗

PubMed2000

[Effects of adenovirus-mediated p16 and p53 genes transfer on apoptosis and cell cycle of lung carcinoma cells].

OBJECTIVE: To explore the synergistic inhibition effect and apoptosis induction of p16 and p53 genes on lung carcinoma cells. METHODS: E1-deficient and replication-defective recombinant p16 and p53 adenoviruses were generated by liposome-mediated co-transfection of recombinant plasmid pAdCMV-p16 or pAdCMV-p53 along with pJM17 and homologous recombination in 293 packaging cell. The lung cancer cell line H358, which had a homozygous deletion of p53 gene and no expression of p16 mRNA and protein, was infected with recombinant p16 and p53 adenovirus either individually or together. RESULTS: Immunohistochemical analysis showed that recombinant adenovirus could transfer p53 gene into tumor cell with 98% efficiency. Western blot indicated that p16 and p53 proteins were expressed at a high level in infected H358 cell. Inhibition effect of p53 gene on proliferation of H358 cell was weaker than that of p16 gene, and the combined use of both genes could completely prevent the proliferation of H358 cell. In situ end-labeling and flow cytometry indicated that p16 could result in G(1) arrest of cell cycle and did not induce H358 cells to undergo apoptosis; p53 also induced apoptosis of few cells besides G(1) arrest; and the simultaneous use of p16 and p53 genes could induce marked apoptosis of H358 cells. CONCLUSION: p16 and p53 genes possess the synergistic inhibiting effect on growth of lung cancer cells and can cooperate to induce apoptosis of H358 cell. The combined application of recombinant p16 and p53 adenoviruses can be used as a new strategy for cancer gene therapy.

Adenoviridae↗

PubMed2000

[Determination of ultra-trace rare-earth elements in human plasma by inductively coupled plasma mass spectrometry].

Inductively coupled plasma mass spectrometry (ICP-MS), highly sensitive inorgnic analytic technique, fits to determine ultra-trace rare-earth elements in human plasma. Under the optimized conditions detection limits for 15 rare-earth elements are in the range of 0.7 (for Eu)-5.4 (for Gd) ng.L-1. Indium as an internal standard element is used to compensate for matrix suppression effect and sensitivity drift. Three kinds of preparation methods, diluted with 1% HNO3, digested with HNO3-H2O2 and with HNO3-HClO4, are checked and compared, and the former is the simplest way to be measured. The samples diluted with 1% HNO3, stored in 4 degrees C, are very steady for 16 days. With the method, 11 healthy plasma samples in Changchun area of China are analysed.

Europium↗

PubMed1999

Multiple mutations of the p53 gene in human mammary carcinoma.

Alteration of the p53 tumor suppressor gene is the most common genetic abnormality in human cancer. In breast cancer, depending on the stage of disease and method of detection, mutation rates of 25-60% have been observed. Multiple mutations of p53 gene in the same tumor however, are rarely reported. In this study we explored the frequency of multiple mutations of p53 gene in mammary carcinoma in a cohort of south Florida patients. Three hundred eighty-four cases of primary breast cancer diagnosed between 1984 and 1986 at the University of Miami, Jackson Medical Center were subjects of this study. Sequence analysis of exons 5 through 8 of p53 was performed on cloned PCR-amplified DNA of formalin-fixed, paraffin-embedded tumors. Two hundred thirty-four of 384 breast cancers (61%) had p53 mutation. Of those, 36 tumors showed more than one mutation; 31 tumors had two mutations, three showed three, one tumor had five mutations, and one case carried six mutations. The majority of mutations were missense (43) followed by silent (35); and most occurred within a single exon. Our study suggests that multiple mutations of p53 suppressor gene in breast cancer are more common than currently believed.

Breast Neoplasms↗

PubMed1999

Epoxomicin, a potent and selective proteasome inhibitor, exhibits in vivo antiinflammatory activity.

The proteasome regulates cellular processes as diverse as cell cycle progression and NF-kappaB activation. In this study, we show that the potent antitumor natural product epoxomicin specifically targets the proteasome. Utilizing biotinylated-epoxomicin as a molecular probe, we demonstrate that epoxomicin covalently binds to the LMP7, X, MECL1, and Z catalytic subunits of the proteasome. Enzymatic analyses with purified bovine erythrocyte proteasome reveal that epoxomicin potently inhibits primarily the chymotrypsin-like activity. The trypsin-like and peptidyl-glutamyl peptide hydrolyzing catalytic activities also are inhibited at 100- and 1,000-fold slower rates, respectively. In contrast to peptide aldehyde proteasome inhibitors, epoxomicin does not inhibit nonproteasomal proteases such trypsin, chymotrypsin, papain, calpain, and cathepsin B at concentrations of up to 50 microM. In addition, epoxomicin is a more potent inhibitor of the chymotrypsin-like activity than lactacystin and the peptide vinyl sulfone NLVS. Epoxomicin also effectively inhibits NF-kappaB activation in vitro and potently blocks in vivo inflammation in the murine ear edema assay. These results thus define epoxomicin as a novel proteasome inhibitor that likely will prove useful in exploring the role of the proteasome in various in vivo and in vitro systems.

Animals↗

PubMed1999

Oxidation of glycosphingolipids under basic conditions: synthesis of glycosyl "serine acids" as opposed to "ceramide acids". Precursors for neoglycoconjugates with increased ligand binding affinity.

Two types of oxidative cleavage of the double bond of glycosphingolipids (GSLs) are described. Oxidation of peracetylated GSL precursors with stoichiometric proportions of KMnO4 and an excess of NaIO4, in a neutral aqueous tert-butanol solvent system, gave nearly quantitative yields of the glycosyl ceramide acid, 2-hydroxy-3-(N-acyl)-4-(O-glycosyl)oxybutyric acid [Mylvaganam, M., and Lingwood, C. A. (1999) J. Biol. Chem. 274, 20725-20732]. However, if the reaction medium was made alkaline, the hydroxyallylic function of the sphingolipid, as a whole, was oxidized and the glycosyl serine acid, 2-(N-acyl)-3-(O-glycosyl)oxypropionic acid, was obtained in good yield. This represents a new type of oxidation reaction. Optimized conditions gave glycosyl ceramide or serine acids with greater than 90% selectivity and in good yields (90%). Oxidation of dGSLs gave serine and ceramide oligosaccharides, devoid of hydrocarbon chains. An intriguing glycosyl species containing 5-hydroxy-4-oxo-3-hydroxy-2-(N-acyl)sphingosine (hydroxy-acyl intermediate) was identified via ESMS analyses. We propose that further oxidation of this intermediate is pH-dependent and will be oxidized to either serine or ceramide acids. On the basis of MS-MS analysis of specific homologues of serine and ceramide acids, two types of collision-induced dissociation (CID) patterns have been established. These CID patterns were then used in the identification of serine and ceramide acids synthesized from natural GSL samples. Also, on a qualitative basis, this oxidation protocol, in conjunction with ESMS, provides a novel method for characterizing the aglycone composition (acyl chain length, unsaturation position, dihydrosphingosine content, etc.) of natural GSLs. A novel class of neohydrocarbon conjugates were synthesized by coupling the acids to rigid hydrocarbon frames such as 2-aminoadamantane. Preliminary studies with conjugates derived from globotriaosyl ceramide (Gb3C), lactosyl ceramide (LC), and galactosyl ceramide (GalC) bound verotoxin with the expected specificity but with affinities much greater than that of the natural glycolipid. Also, the ceramide acid-based conjugates were better ligands than serine acid conjugates.

Acylation↗

PubMed1999

Total synthesis of the potent proteasome inhibitor epoxomicin: a useful tool for understanding proteasome biology.

Epoxomicin (1), a peptide alpha',beta'-epoxyketone isolated from the actinomycete strain No.Q996-17, possesses potent in vivo anti-tumor and anti-inflammatory activities. In this paper, we report the first syntheses of epoxomicin, [3H]-epoxomicin, and a biotinylated epoxomicin analog as well as the absolute configuration of the epoxide stereocenter. The natural product and derivatives have permitted the first identification of the proteasome as the specific cellular target of epoxomicin.

Animals↗

PubMed1999

Eponemycin exerts its antitumor effect through the inhibition of proteasome function.

Cell cycle progression requires the proteasome-mediated degradation of key regulatory proteins such as cyclins, cyclin-dependent kinase inhibitors, and anaphase-inhibitory proteins. Given the central role of the proteasome in the destruction of these proteins, proteasome inhibition has been proposed as a possible cancer therapy. We report here that dihydroeponemycin, an analogue of the antitumor and antiangiogenic natural product eponemycin, selectively targets the 20S proteasome. Dihydroeponemycin covalently modifies a subset of catalytic proteasomal subunits, binding preferentially to the IFN-gamma-inducible subunits LMP2 and LMP7. Moreover, the three major peptidolytic activities of the proteasome are inhibited by dihydroeponemycin at different rates. In addition, dihydroeponemycin-mediated proteasome inhibition induces a spindle-like cellular morphological change and apoptosis. These results validate the proteasome as a target for antitumor pharmacological intervention and are relevant for the design of novel chemotherapeutic strategies.

Amides↗

PubMed1999

Synthesis, structural characterization and antitumor activity evaluations of copper complex with tetraazamacrocyclic ligand.

Cu (II) complex with 1,4,7,10-tetrakis(2-cyanoethyl-)-1,4,7,10-tetraazacyclododecane was prepared and characterized by X-ray diffraction. Four nitrogen atoms of macrocyclic ligand and oxygen atom of water molecule defined a tetragonal pyramidal polyhedron surrounding the central copper atom. Preliminary pharmacological tests showed that it had antitumor activity against P388 and BEL-7404 cell lines in vitro. Also it exhibited perturbation effects to K562 tumor cell lines at G0-G1 stage and further studies showed that it can cleave supercoiled DNA (pBR 322) to nicked and linear DNA in aerobic condition.

Antineoplastic Agents↗

PubMed1999

Lifestyle factors and chronic diseases: application of a composite risk index.

BACKGROUND: Assessing a combination of modifiable lifestyle practices may be a practical tool to modify patients' health behavior in counseling. Therefore, we developed a chronic disease risk index (CDRI) and investigated its relation with chronic disease in a multiethnic cohort. METHODS: A total of 15,693 men and 16,007 women in Hawaii who reported their diet and other lifestyle behaviors between 1975 and 1980 were followed until 1994. A semi-quantitative composite CDRI with scores ranging from 1 to 10 included the rankings for smoking, alcohol use, body mass index, fat intake, and fruit and vegetable consumption. Cox's proportional hazards regression was used to estimate the relative risk for chronic diseases. RESULTS: When comparing the highest to the lowest CDRI category, the respective relative risks (RRs) for total mortality were 2.9 (95% CI 2.3-3.8) and 3.8 (95% CI 2.9-5.0) for men and women. With higher CDRIs, the RRs for cancer incidence, mortality from cancer, coronary heart disease, and stroke increased significantly. Among the five components of the CDRI, smoking had the greatest influence on chronic disease risk, followed by body mass index. CONCLUSIONS: Positive health behavior reflected by the CDRI is associated with a lower risk of cancer and with greater longevity.

Adult↗

PubMed1999

In vitro maturation of human preovulatory oocytes reconstructed by germinal vesicle transfer.

OBJECTIVE: To describe a micromanipulation-electrofusion procedure for transferring germinal vesicles (GVs) between immature human oocytes. DESIGN: Pilot study to assess oocyte maturation after an invasive micromanipulation procedure. SETTING: Research laboratory at a university medical center. PATIENT(S): Immature oocytes were discarded from intracytoplasmic sperm injection (ICSI)-IVF cycles of patients 23-48 years of age. INTERVENTION(S): Initially, GV removal and transfer were performed on the same oocyte; these "self-reconstructed" oocytes were then cultured in vitro for up to 50 hours and examined periodically for maturation as judged by the extrusion of the first polar body. In a second study, GVs from oocytes of "old" patients (>38 years old) were successfully transferred into enucleated immature oocytes of "young" patients (<31 years old). MAIN OUTCOME MEASURE(S): Extrusion of the first polar body was monitored in "reconstructed" and control oocytes; karyotypes also were analyzed at meiosis II. RESULT(S): From 48 oocytes from old patients, 12 GVs were successfully removed, transferred, and fused into previously enucleated oocytes from young patients. After in vitro culture, 7 of these "reconstructed" oocytes matured to meiosis II, a maturation rate not significantly different from that observed in nonmanipulated controls. A normal, second meiotic metaphase chromosome complement was observed in 4 of 5 reconstructed oocytes. CONCLUSION(S): Normal meiosis can occur after the transfer of a GV into an enucleated host oocyte. Germinal vesicle transfer may be a valuable research procedure that generates cell models to characterize the cytoplasmic-nuclear interplay for cell cycle regulation, maturation, and fertilization in the human oocyte; it also may be a potentially attractive alternative to oocyte donation.

Adult↗

PubMed1999

Nuclear transfer in the rhesus monkey: practical and basic implications.

In early 1997, the birth of a lamb after transfer of the nucleus from an adult mammary gland cell into an enucleated oocyte, along with the production of rhesus monkeys by nuclear transfer of embryonic cells, marked a reemergence of the field of mammalian cloning. Clonally derived rhesus monkeys would be invaluable in biomedical research, and the commercial interests in transgenic sheep and cattle propagated by cloning are substantial. Nuclear transfer technology is under consideration in human in vitro fertilization clinics to overcome infertility secondary to advanced maternal age or mitochondrial-based genetic disease. Nuclear transfer involves preparing a cytoplast as a recipient cell, in most cases a mature metaphase II oocyte from which the chromosomes have been removed. A donor nucleus cell is then placed between the zona and the cytoplast, and fusion, as well as cytoplast activation, is initiated by electrical stimulation. Successful reprogramming of the donor cell nucleus by the cytoplast is critical--a step that may be influenced by cell cycle stage. Embryos produced by nuclear transfer are cultured in vitro for several cell divisions before cryopreservation or transfer to the oviduct or uterus of a host mother. The efficiency of producing live young by nuclear transfer in domestic species is low, with a high frequency of developmental abnormalities in both preterm and term animals. However, a number of pregnancies have now been established using fetal cells as the source of donor nuclei. The use of cell lines not only allows large clone sizes but also supports the ability to genetically manipulate cells in vitro before nuclear transfer. Ongoing research focused on the production of clonally derived rhesus monkeys using fetal fibroblasts and embryonic stem cells as the source of donor nuclei will be reviewed.

Animals↗

PubMed1999

A pilot study of mammographic density patterns among Japanese women.

Mammographic density patterns, which refer to the distribution of fat, connective, and epithelial tissue in the healthy female breast, have been shown to be related to breast cancer risk. We used a quantitative method to assess mammographic densities in 41 mammograms from women in Japan without a diagnosis of breast cancer. Information about reproductive behavior and family history for breast cancer was available from a questionnaire. The statistical analysis applied Spearman correlation coefficients and multiple linear regression. The breast size as measured on the cranio-caudal view of the mammogram was approximately 12% larger, the size of the dense areas was 20% smaller, and the mean percent mammographic densities were 30% greater among premenopausal than among postmenopausal women. We found a strong relation between age at menarche and mammographic densities in premenopausal women and significant associations for age, family history of breast cancer, and age at menopause with mammographic densities in postmenopausal women. These preliminary data will be used to plan a future study that will compare mammographic density patterns and the relative importance of dietary, reproductive, and anthropometric factors between women in Japan and in the United States.

Adult↗

PubMed1999

Frequency and pattern of p53 gene mutation in a cohort of Spanish women with node-negative breast cancer.

Ethnic, racial and regional differences in the frequency and pattern of p53 gene mutations have been well documented. Some of these differences have been shown to have an impact on the survival of patients with breast cancer. In this study we explored the frequency and pattern of p53 abnormality in a cohort of Spanish women with node-negative breast cancer using PCR, subcloning and DNA sequencing of archival tumors. One hundred and seventy-eight cases of breast cancer diagnosed between 1981 and 1986 at the University of Oviedo Hospital in Oviedo, Spain were subjects of this study. Sequence analysis of exons 5 through 8 of p53 was performed on subcloned PCR-amplified DNA, extracted from formalin-fixed, paraffin-embedded tumors. Appropriate positive, negative, PCR, and polymerase controls were utilized and evaluated. Duplicate samples of the genomic DNA were re-evaluated on all cases showing more than one mutation. One hundred and five out of 178 breast cases (59%) carried one or more p53 gene mutations. Mutations were distributed randomly from codon 128 to 305. There were 123 (88%) transition, 10 (7%) transversion, 5 (3.5%) splice junction mutations, and 2 (1.5%) deletions. Eighty-three cases (61.5%) had missense mutation, 45 (33.5%) silent, 5 (3.5%) nonsense and 2 (1.5%) frameshifts. Eighty (75%) of 120 transitions were G:C to A:T, 11 (25%) of which occurred at CpG sites. Sixteen mutations were in novel codons not reported in breast cancers previously. Codons with the highest frequency of mutations in this group were 278, 273, 213 and 227. We also detected 27 tumors with more than one mutation within a single exon or in different exons in the same patient. These findings suggest that the frequency and pattern of p53 mutations in this group of Spanish women with breast cancer is different than those reported in the United States and Northern Europe.

Breast Neoplasms↗

PubMed1999

The expression of p53 tumor suppressor gene in breast cancer cells is down-regulated by cytokine oncostatin M.

Previously (J. Liu, et al., Cell Growth Differ., 8: 667-676, 1997), we showed that oncostatin M (OM), a cytokine produced by activated T cells and macrophages, inhibited the proliferation of breast cancer cells derived from solid tumors and malignant effusions. OM-treated cells showed reduced growth rates and differentiated phenotypes. Because the p53 tumor suppressor protein plays an important role in cellular proliferation, we examined p53 protein expression in three OM-responsive breast cancer cell lines, MCF-7, MDA-MB231, and H3922. Western blot analysis showed that p53 protein levels in all three of the cell lines were decreased by OM treatment. Reduction of p53 protein was detected after 1 day of OM treatment and reached maximal suppression of 10-20% of control after 3 days in H3922 and 40% of control after 4 days in MCF-7 cells. A comparison of p53 mRNA in OM-treated cells versus untreated control cells showed that exposure to OM reduced the steady-state levels of p53 mRNA transcripts to an extent similar to that of the p53 protein levels. This observation suggests that the effect of OM on p53 protein expression does not occur at the posttranslational level. Nuclear run-on assays verified that OM decreased the number of actively transcribed p53 mRNAs, which suggests a transcriptional regulatory mechanism. The effect of OM on p53 expression seems to be mediated through the extracellular signal-regulated kinase (ERK) pathway, inasmuch as the inhibition of ERK activation with a specific inhibitor (PD98059) to the ERK upstream kinase mitogen/extracellular-regulated protein kinase kinase abrogated the OM inhibitory activity on p53 expression in a dose-dependent manner. In addition to OM, we showed that the p53 protein expression in MCF-7 cells was also decreased by phorbol 12-myristate 13-acetate treatment (PMA). Because both OM and PMA induce MCF-7 cells to differentiate, our data suggest that p53 expression in breast cancer cells is down-regulated during the differentiation process.

Blotting, Western↗

PubMed1999

Exploring the feasibility and effects of a high-fruit and -vegetable diet in healthy women.

Based on reports that fruits and vegetables may protect against breast cancer, this randomized intervention study tested the feasibility of increasing fruit and vegetable intake among healthy women to 9 daily servings through individual dietary counseling and group activities. Adherence to the dietary recommendations was monitored by 24-h food recalls, log sheets, and plasma carotenoid assessments. To explore possible cancer protective mechanisms of fruits and vegetables, we investigated the treatment effect on plasma phenol levels and on thiobarbituric acid-reactive substances measured as malondialdehyde equivalents, a possible marker of oxidative damage. At baseline, women in the intervention (n = 13) and control (n = 16) group reported an average daily consumption of 3.3 and 3.2 fruit and vegetable servings, respectively. After 3 and 6 months of intervention, intake in the intervention group had increased to 8.3 and 7.4 servings, whereas the control group reported an average of 4.2 and 4.1 daily servings. An increase of plasma carotenoid levels from 1249 microg/liter at baseline to 1854 and 1827 microg/liter after 3 and 6 months confirmed compliance with the dietary recommendations in the intervention group. Plasma carotenoid levels among controls changed slightly from 1165 to 1231 and 1291 microg/liter Whereas total phenol levels did not respond according to our hypothesis, malondialdehyde levels decreased slightly in the intervention group. These results suggest that motivated women can substantially increase their fruit and vegetable intake, which leads to a notable increase in plasma carotenoid levels.

Adult↗