Association between the CYP1A1 polymorphisms and hepatocellular carcinoma: a meta-analysis.

Numerous studies have evaluated the association between CYP1A1 gene polymorphisms and hepatocellular carcinoma (HCC) risk. However, the specific association is still controversial. The aim of our study was to clarify the effects of CYP1A1 gene polymorphisms (3801 T＞C and A2455G) on HCC risk by conducting a meta-analysis. We conducted searches of the literature published in PubMed and EMBASE databases up to April 2014. We estimated the pooled odds ratio with its 95% confidence interval to assess the association using a fixed or random-effects model. Publication bias was investigated by the Begg funnel plot. Meta-analysis was performed using the STATA package version 12.0. Meta-analysis results showed no significant association between the CYP1A1 3801 T＞C polymorphism and HCC risk. In a subgroup analysis by nationality, we found a significant association between 3801 T＞C polymorphism and HCC risk in Asians (TT vs TC: OR = 0.77, 95%CI = 0.60-0.99). As for A2455G, the meta-analysis indicated no significant association between the CYP1A1 A2455G polymorphism and HCC risk. In conclusion, the 3801 T＞C polymorphism in the CYP1A1 gene may be related to increased risk of HCC in Asians. Conclusive evidence on the effects of the variants in HCC should be addressed in further studies.

How Dietary Patterns are Related to Inflammaging and Mortality in Community-Dwelling Older Chinese Adults in Hong Kong - A Prospective Analysis.

INTRODUCTION: Studies examining dietary patterns and inflammageing in relation to mortality are limited. OBJECTIVE: We examined the influence of various dietary patterns on all-cause and cardiovascular disease (CVD) mortality, taking into account demographics, lifestyle factors, and serum inflammatory markers. METHODS: We conducted multivariate Cox regression analyses using data from a cohort of community-dwelling older Chinese adults (1,406 men, 1,396 women) in Hong Kong. Baseline interviewer administered questionnaires covered dietary intake estimation and dietary pattern generation from the food frequency questionnaire, demographic and lifestyle factors, cognitive function and depressive symptoms. Serum high-sensitivity C-reactive protein (hsCRP) and 25-hydroxyvitamin D (25OHD) were measured. All-cause and CVD mortality data at 14-year follow up were retrieved from an official database. RESULTS: In men, higher hsCRP level was associated with lower Diet Quality Index-International (DQI-I) score, Mediterranean-DASH Intervention for Neurodegenerative Delay Diet (MIND) score, Okinawan diet score, "vegetables-fruits" pattern score and "snacks-drinks-milk" pattern score. Higher serum 25OHD level was associated with higher Mediterranean Diet Score (MDS) but lower "snacks-drinks-milk" pattern score. None of the dietary pattern scores was associated with all-cause or CVD mortality after adjusting for all covariates. In women, hsCRP level and serum 25OHD level were not associated with any dietary patterns. Higher DQI-I score (HR=0.77 (95% CIs: 0.59, 0.99) highest vs. lowest tertile, p-trend=0.038) and Okinawan diet score (HR=0.78 (95% CIs: 0.61, 1.00) highest vs lowest tertile, p-trend=0.046) was associated with a lower risk of all-cause mortality, whereas higher MIND score (HR=0.63 (95% CI: 0.36, 1.09) highest vs. lowest tertile, p-trend=0.045) was associated with a reduced risk of CVD morality in the multivariate adjusted model. CONCLUSION: Higher DQI-I score and Okinawan diet score were associated with a lower risk of all-cause mortality, and higher adherence to the MIND diet was related to a reduced risk of CVD mortality in community-dwelling Chinese older women.

Influence of Dietary Patterns and Inflammatory Markers on Atherosclerosis Using Ankle Brachial Index as a Surrogate.

OBJECTIVE: To examine the influence of various dietary patterns on ankle-brachial index (ABI) as a surrogate of atherosclerosis, taking into account serum levels of C-reactive protein (CRP) and factors that predispose to inflammation and/or endothelial damage (homocysteine) or may be protective (tryptophan, vitamin D), as well as age, gender, and lifestyle risk factors. DESIGN: Cross sectional analysis. SETTING: Cohort of 4000 men and women aged 65 years living in the community in Hong Kong SAR China. MEASUREMENTS: Interviewer administered questionnaire that includes dietary intake estimation by the food frequency records (from which various dietary patterns can be characterized), socioeconomic status, smoking habit, alcohol consumption, physical activity. Clinical measurements include body weight and height, and ankle-brachial index using hand-held Doppler machine. Laboratory measurements include assays of serum 25 hydroxyvitamin D, CRP, homocysteine, and tryptophan using LC/MC methods. RESULT: Compared with the lowest quintile of vitamin D level, higher quintile was associated with lower prevalence of ABI <0.9 in women only after multiple adjustments of covariates. In men the low score group of the vegetables-fruits dietary pattern was associated with increased odds of having ABI<0.9 after adjusting for all covariates (OR=16.51 (95% CI: 2.21, 123.26)). Similar findings apply to the low score group in the MIND (Mediterranean-DASH Intervention for Neurodegenerative Delay Diet) pattern (OR=2.6 (95% CI: 1.24, 5.42)). In women, ABI<0.9 was associated with low Dietary Quality Index (DQI) score (OR=2.05 (95% CI: 1.22, 3.43)), and low score group of snacks-drinks-milk products dietary pattern (OR=3.07 (95% CI: 1.05, 9.04)). No association was observed for the Mediterranean Dietary Pattern (MDS), Dietary Approaches to Stop Hypertension (DASH), Okinawan and "meat-fish" dietary patterns in either men or women. CONCLUSION: We conclude that any dietary patterns emphasizing fruit and vegetable intake are associated with better vascular health using ABI as an indicator.

Modified miniprep method for the rapid recovery of episomes from transfected breast epithelial cells.

Episomal vectors such as pCEP4 are useful in expression cloning because they can replicate in both prokaryotes and eukaryotic cells. We have found a rapid and efficient means of extracting them from transfected MCF-10A nonmalignant human breast epithelial cells. We show that a plasmid miniprep protocol, modified by the addition of an extraction that eliminates a DNase activity, can consistently harvest pCEP4 episomes from the transfected cells (516 +/- 112 pg/harvest, mean +/- standard deviation; n = 11). The quality of the episomal DNA obtained in this manner was verified by PCR, Southern blot and the retransformation of Escherichia coli. This simple method enables the efficient recovery of episomes and is applicable in the expression cloning of potential oncogenes using host MCF-10A cells.

Multiple factors other than p53 influence colon cancer sensitivity to paclitaxel.

PURPOSE: To determine factors which influence the sensitivity of human colorectal carcinoma cell lines to paclitaxel. METHODS: The paclitaxel sensitivity of ten human colorectal carcinoma cell lines, and a panel of RKO colon carcinoma cell lines, isogenic except for p53 status, were studied. The inhibitory concentrations causing a 50% decrease in growth (IC50) were assayed after 3, 24, and 96 h after paclitaxel exposure. The doubling time (DT) and cell cycle parameters of cells were also measured. The expression of the multidrug resistance glycoprotein-1 (MDR-1), bcl-2 and bax was quantitatively assessed by immunoblotting. RESULTS: Mean IC50 values at 24 and 96 h drug exposure were about 1.5 logs lower than the IC50 values at 3 h, regardless of the p53 status. No difference was found between the IC50 values of wild-type and mutant p53 cells, or among the RKO panel of cells. Correlation analysis showed that: (1) resistance was associated with longer DTs, but this was generally abated by a 96-h exposure; (2) with a 3-h exposure, the combination of MDR, bcl-2 and bax parameters with DT (DT + MDR + bcl-2 bax) best correlated with IC50 values (r = 0.77); (3) with a 96-h exposure, in spite of the generally decreased IC50 values, a combination of MDR-1, bcl-2 and bax parameters (MDR + bcl-2-bax) best correlated with the IC50 values (r = 0.71). CONCLUSIONS: These results suggest that the exposure duration, DT, and expression of MDR-1, bcl-2 and bax each contribute to paclitaxel sensitivity of human colorectal carcinoma cells. In assessing paclitaxel drug resistance, multiple factors should always be considered. There may be a therapeutic window for taxanes in colon cancer by optimizing pharmacokinetics and modulating MDR-1 and bcl-2 resistance factors.

Phosphorothioated antisense c-myc oligonucleotide inhibits the growth of human colon carcinoma cells.

BACKGROUND: The functional significance of c-myc overexpression in human colon carcinoma is unclear. MATERIALS AND METHODS: Three human colon carcinoma cell lines, LS174T, SW1116 and SW48, were treated in vitro with phosphorothioate modified antisense oligonucleotides (ASO), complementary to the c-myc translation initiation site, or two control oligonucleotides. Growth was assayed by the methyl tetrazolium (MTT) assay and colony formation. C-myc, retinoblastoma (Rb), carcinoembryonic antigen (CEA), and alkaline phosphatase (AP) expression was assessed by immunoblotting. RESULTS: The ASO specifically inhibited growth of all three human colon carcinoma cell lines. Further studies were conducted on LS174T because growth inhibition was associated with a more differentiated morphology. In LS174T cells, ASO caused a dose dependent decrease in c-myc and Rb protein expression, but an increase in CEA and AP expression. CONCLUSIONS: The growth of human colon carcinoma cells is dependent on c-myc expression, and the inhibition of c-myc expression in LS174T is associated with a more differentiated phenotype.

The effects of metabotropic glutamate receptor 7 allosteric agonist N,N'-dibenzhydrylethane-1,2-diamine dihydrochloride on developmental sevoflurane neurotoxicity: role of extracellular signal-regulated kinase 1 and 2 mitogen-activated protein kinase signaling pathway.

The present study was designed to evaluate the possible neuroprotective effects of metabotropic glutamate receptor (mGluR7) allosteric agonist N,N'-dibenzhydrylethane-1,2-diamine dihydrochloride (AMN082) on developmental sevoflurane neurotoxicity. To achieve the objective, hippocampal cultures (7 DIV, 7 day in vitro) were treated with different doses of L-(+)-2-amino-4-phosphonobutyric acid (L-AP4, an agonist of group III mGluRs), (RS)-α-Methylserine-O-phosphate (MSOP, an antagonist of group III mGluRs), AMN082 or cis-2-[[(3,5-dichlorophenyl)amino]carbonyl]cyclohexanecarboxylic acid (VU0155041, an agonist of mGluR4) before exposed to sevoflurane. Cell apoptosis were determined by flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL)-staining. For in vivo study, rat pups (7 PND, 7 postnatal day) were injected with AMN082, L-AP4 or saline before sevoflurane exposure. Extracellular signal-regulated kinase 1 and 2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38, caspase-3, Bcl-2, and Bax were detected by Western blot. The locomotor activity and cognitive functions were evaluated by open-field test and Morris water maze (MWM), respectively. We found that L-AP4 prevented sevoflurane-induced cell apoptosis, but MSOP promoted. Specially, application of AMN082 contributed to the relief of sevoflurane-induced apoptosis in vitro, whereas VU0155041 did not. In addition, sevoflurane treatment led to a decrease of Bcl-2 and an increase of caspase-3 and Bax, which were mitigated by AMNO82 in vivo. Moreover, we showed that sevoflurane treatment resulted in a remarkable suppression of phospho-ERK1/2, which was restored by AMN082. Application of U0126 (an inhibitor of MEK) abolished the neuroprotective effects of AMN082 on sevoflurane neurotoxicity both in vitro and in vivo. In addition, sevoflurane exposure also led to an increase of phospho-JNK, but SP600125 (an inhibitor of JNK) did not attenuate sevoflurane-induced apoptosis. The total and phosphorylated p38 remained unchanged in sevoflurane-treated rat pups. Finally, AMN082 improved the learning and memory defects caused by postnatal sevoflurane exposure without alternations in emotion or locomotor activity. These preliminary data indicate that AMN082 may protect immature brain against sevoflurane neurotoxicity, and the ERK1/2 MAP kinase signaling is likely to be involved. Further studies are needed to fully assess the neuroprotective role of mGluR7 agonist AMN082 in developmental anesthetic neurotoxicity.

The effect of timing of application of positive end-expiratory pressure on oxygenation during one-lung ventilation.

Many studies have confirmed that applying positive end-expiratory pressure (PEEP) to the dependent lung during one-lung ventilation (OLV) improves oxygenation. Our purpose was to investigate the best time and level of PEEP application. Thirty patients undergoing thoracic surgery were randomised into three groups. After 20 minutes of two-lung ventilation (TLV) in the lateral position, all patients received OLV for one hour During OLV, 0, 5, 10 cmH2O PEEP were applied in order in group A, with each level sustained for 20 minutes. Group B had 5 cmH2O PEEP applied and maintained for one hour Patients in group C received PEEP with levels set in the opposite order to that of group A. The ventilation model was then converted to TLV. PaO2, PaCO2 and respiratory mechanical variables were compared at five different time points among groups, 20 minutes after TLV (T1), 20 (T2), 40 (T3) and 60 minutes (T4) after OLV and 20 minutes after conversion to TLV (T5). We found that PaO2 was lower in group A than the other two groups at T2 (P <0.05). PaO2 decreased significantly at T5 compared with T1 (P <0.05) in group A only. When PEEP was set to 10 cmH2O, the airway pressure increased significantly (P <0.05). These findings indicate that PEEP applied at the initial time of OLV improves oxygenation most beneficially. Five cmH2O PEEP may produce this beneficial effect without the increase in airway pressure associated with 10 cmH2O PEEP.

Somatic mutations in c-myc intron I cluster in discrete domains that define protein binding sequences.

The activated c-myc allele in Burkitt's lymphoma tumor cells is associated with a clustering of somatic mutations within intron I near the exon I boundary. We have identified several discrete protein binding sites within this region of c-myc intron I designated as myc intron factor-1 (MIF-1), MIF-2, and MIF-3. In addition to our previous characterization of a 20-nucleotide binding site for MIF-1, we now have identified adjacent 20-nucleotide and 34-nucleotide binding sites for MIF-2 and MIF-3, respectively. All three elements are protected from exonuclease digestion by nuclear protein extracts, and each gives rise to a distinct migration pattern on mobility shift assays. In addition, MIF-1, 2, and 3 share a 5-nucleotide (TTATG) internal sequence, which may account for cross-competition of these binding sites in the exonuclease protection experiment. Deletion mutant analyses showed that selective removal of the MIF-3 binding site alone was sufficient to enhance chloramphenicol acetyltransferase reporter activity similar to that observed with larger deletions of myc intron I. We have demonstrated that somatic mutations in activated c-myc alleles are frequently clustered in discrete domains that define protein recognition sequences.

Cytotoxic oxoisoaporphine alkaloids from Menispermum dauricum.

Four new oxoisoaporphine alkaloids, daurioxoisoporphines A-D (1-4), were isolated from the rhizomes of Menispermum dauricum. The structures of these alkaloids were established by spectroscopic methods. The cytotoxic evaluation of 1 and 2 is reported against four cancer cell lines.