The purpose of this study was to identify microRNAs (miRNAs) involved in the pathology of colorectal cancer (CRC) liver metastasis and investigate their underlying mechanisms. A total of 39 miRNAs ...were identified to be differentially expressed between 16 primary CRC tissues with liver metastases and 16 CRC tissues without liver metastases from 32 patients by Affymetric miRNA microarrays. A panel of eight miRNAs were confirmed to be significantly and differentially expressed between CRC tissues with and without liver metastases through quantitative reverse‐transcription polymerase chain reaction (RT‐PCR) analysis in the 32 patients. In a validated cohort of 99 CRC patients (44 with and 55 without liver metastases), only miR‐214 was validated to be significantly down‐regulated in CRC with liver metastases, which was associated with an unfavorable prognosis. Ectopic expression of miR‐214 suppressed proliferation, migration, and invasion in vitro, tumor growth and liver metastasis in an in vivo xenograft mouse model, whereas miR‐214 knockdown promoted proliferation, migration, and invasion in CRC cell lines. Further studies indicated that fibroblast growth factor receptor 1 (FGFR1) was a potential target of miR‐214. Restoring miR‐214 expression in CRC cells decreased endogenous FGFR1 messenger RNA (mRNA) and protein levels. FGFR1 knockdown mimicked the tumor suppressive effect of miR‐214 on CRC cells, while reintroduction of FGFR1 abolished the tumor suppressive effect of miR‐214 on CRC cells. Moreover, miR‐214 expression levels were inversely correlated with FGFR1 in CRC patients. Conclusion: Down‐regulation of miR‐214 expression was correlated with increased FGFR1 expression levels, which may contribute to increased CRC liver metastasis. miR‐214 may serve as a potential marker to predict survival, and the miR‐214‐FGFR1 axis may be a therapeutic target in CRC patients. (Hepatology 2014;60:598–609)
Objective： To assess the short-term effect of scaling and root planing （SRP） and essential-oils mouthwash on the levels of specific bacteria in Chinese adults. Methods： Fifty Chinese adults with ...chronic periodontitis were randomly assigned to full-mouth SRP or a 7-d essential-oils mouthwash regimen. In addition, 22 periodontally healthy adults used essential-oils mouthwash for 7 d. Clinical examination and plaque/saliva sampling were performed at baseline and on Day 7. Quantitative real-time polymerase chain reaction （PCR） was used to measure Aggregatibacter actinomycetemcomitans （Aa）, Fusobacterium nucleatum （Fn）, Porphyromonas gingivalis （Pg）, Prevotella intermedia （Pi）, and total bacterial loads in saliva, supra- and sub-gingival plaque samples. Results： The detection frequencies of four tested species remained unchanged after either treatment. However, the bacterial loads of Fn, Pg, and Pi were significantly reduced by SRP; the mean reduction of bacterial counts in saliva ranged from 52.2% to 62.5% （p〈0.01）, in supragingival plaque from 68.2% to 81.0% （p〈0.05）, and in subgingival plaque from 67.9% to 93.0% （p〈0.01）. Total bacterial loads were reduced after SRP in supra- and sub-gingival plaque （p〈0.05）. Essential-oils mouthwash reduced Fn levels in supragingival plaque by a mean of 53.2%, and reduced total bacterial loads in supra- and sub-gingival plaque （p〈0.01）. In subgingival plaque from periodontal patients, Pg and Pi reductions were high after SRP compared to essential-oils mouthwash （93.0% vs. 37.7% and 87.0% vs. 21.0%, p〈0.05）. No significant bacterial reduction was observed in periodontally healthy subjects using essential-oils mouthwash. Conclusions： SRP and essential-oils mouthwash both have an impact on saliva and gingival plaque flora in Chinese periodontitis patients in 7 d, with greater microbiological improvement by SRP.
Light‐assisted Li–CO2 battery, emerging as a powerful battery‐technology to utilize light‐energy, has attracted much attention yet it is still limited by the lack of efficient photocathode catalysts ...toward simultaneously promoted CO2 activation/evolution processes. Here, a kind of phthalocyanine‐based metal–organic framework nanosheet (CoPc–Mn–O) is prepared and applied as efficient photocathode catalyst in light‐assisted Li–CO2 battery. Thus‐produced CoPc–Mn–O shows nanosheet‐morphology (≈1 nm), dual active metal‐sites (i.e., Co–N4 and Mn–O4), high conductivity, and photosensitivity. The resulting battery delivers a high round‐trip efficiency of 98.5% with an ultralow voltage hysteresis of 0.05 V and superior cycling‐stability (81.3%) for 60 h at 0.02 mA cm−2, which is represented to be one of the best photo‐cathodes to date. Density functional theory calculations and characterizations prove that the presence of photosensitive CoPc–Mn–O with dual active metal‐sites endows an activation energy (3.2 eV) for CO2 activation under light condition and simultaneously promote the CO2 evolution to enhance the battery efficiency.
A kind of phthalocyanine‐based metal–organic framework nanosheet (i.e., CoPc–Mn–O) is synthesized and presents nanosheet morphology, high conductivity, dual active metal‐sites (i.e., Co–N4 and Mn–O4 sites) and excellent photosensitivity, which serves as efficient photocathode catalyst for photo‐assisted Li–CO2 battery
Many applications of CRISPR/Cas9-mediated genome editing require Cas9-induced non-homologous end joining (NHEJ), which was thought to be error prone. However, with directly ligatable ends, ...Cas9-induced DNA double strand breaks may be repaired preferentially by accurate NHEJ.
In the repair of two adjacent double strand breaks induced by paired Cas9-gRNAs at 71 genome sites, accurate NHEJ accounts for about 50% of NHEJ events. This paired Cas9-gRNA approach underestimates the level of accurate NHEJ due to frequent + 1 templated insertions, which can be avoided by the predefined Watson/Crick orientation of protospacer adjacent motifs (PAMs). The paired Cas9-gRNA strategy also provides a flexible, reporter-less approach for analyzing both accurate and mutagenic NHEJ in cells and in vivo, and it has been validated in cells deficient for XRCC4 and in mouse liver. Due to high frequencies of precise deletions of defined "3n"-, "3n + 1"-, or "3n + 2"-bp length, accurate NHEJ is used to improve the efficiency and homogeneity of gene knockouts and targeted in-frame deletions. Compared to "3n + 1"-bp, "3n + 2"-bp can overcome + 1 templated insertions to increase the frequency of out-of-frame mutations. By applying paired Cas9-gRNAs to edit MDC1 and key 53BP1 domains, we are able to generate predicted, precise deletions for functional analysis. Lastly, a Plk3 inhibitor promotes NHEJ with bias towards accurate NHEJ, providing a chemical approach to improve genome editing requiring precise deletions.
NHEJ is inherently accurate in repair of Cas9-induced DNA double strand breaks and can be harnessed to improve CRISPR/Cas9 genome editing requiring precise deletion of a defined length.
Abstract
Aims
Circular RNAs (circRNAs) are involved in gene regulation in a variety of physiological and pathological processes. The present study aimed to investigate the effect of circRNA_000203 on ...cardiac hypertrophy and the potential mechanisms involved.
Methods and results
CircRNA_000203 was found to be up-regulated in the myocardium of Ang-II-infused mice and in the cytoplasma of Ang-II-treated neonatal mouse ventricular cardiomyocytes (NMVCs). Enforced expression of circRNA_000203 enhances cell size and expression of atrial natriuretic peptide and β-myosin heavy chain in NMVCs. In vivo, heart function was impaired and cardiac hypertrophy was aggravated in Ang-II-infused myocardium-specific circRNA_000203 transgenic mice (Tg-circ203). Mechanistically, we found that circRNA_000203 could specifically sponge miR-26b-5p, -140-3p in NMVCs. Further, dual-luciferase reporter assay showed that miR-26b-5p, -140-3p could interact with 3′-UTRs of Gata4 gene, and circRNA_000203 could block the above interactions. In addition, Gata4 expression is transcriptionally inhibited by miR-26b-5p, -140-3p mimic in NMVCs but enhanced by over-expression of circRNA_000203 in vitro and in vivo. Functionally, miR-26b-5p, -140-3p, and Gata4 siRNA, could reverse the hypertrophic growth in Ang-II-induced NMVCs, as well as eliminate the pro-hypertrophic effect of circRNA_000203 in NMVCs. Furthermore, we demonstrated that NF-κB signalling mediates the up-regulation of circRNA_000203 in NMVCs exposed to Ang-II treatment.
Conclusions
Our data demonstrated that circRNA_000203 exacerbates cardiac hypertrophy via suppressing miR-26b-5p and miR-140-3p leading to enhanced Gata4 levels.
Graphical Abstract
Graphical Abstract
Abnormal epigenetic regulation has been implicated in oncogenesis. We report here the identification of somatic mutations by exome sequencing in acute monocytic leukemia, the M5 subtype of acute ...myeloid leukemia (AML-M5). We discovered mutations in DNMT3A (encoding DNA methyltransferase 3A) in 23 of 112 (20.5%) cases. The DNMT3A mutants showed reduced enzymatic activity or aberrant affinity to histone H3 in vitro. Notably, there were alterations of DNA methylation patterns and/or gene expression profiles (such as HOXB genes) in samples with DNMT3A mutations as compared with those without such changes. Leukemias with DNMT3A mutations constituted a group of poor prognosis with elderly disease onset and of promonocytic as well as monocytic predominance among AML-M5 individuals. Screening other leukemia subtypes showed Arg882 alterations in 13.6% of acute myelomonocytic leukemia (AML-M4) cases. Our work suggests a contribution of aberrant DNA methyltransferase activity to the pathogenesis of acute monocytic leukemia and provides a useful new biomarker for relevant cases.
Objective： Total hip arthroplasty （THA） and hemiarthroplasty （HA） are effective methods currently used to treat femoral neck fracture in elderly patients, but the two options remain controversial in ...patients over 70 years old. The main purpose of our study was to determine whether THA or HA is a superior treatment of femoral fractures involving a displaced neck in patients who are over 70 years of age. Methods： A computer-based online search of Medline （1970-2011）, PubMed （1977-2011）, and the Cochrane Cen- tral Register of Controlled Trials （2002-201 l） was conducted. Six relevant randomized controlled trials with a total of 739 patients were included for the final analysis. The analysis was performed with software RevMan 5.0. Results： We found that compared with THA, HA needed shorter average time and lost less blood. While over the long-term follow-up, THA patients exhibited significantly less pain and better function and were less likely to require a revision hip surgery. Postoperative infection was equally common among HA and THA patients. Conclusions： The significant differences in outcomes suggest that THA is a valuable treatment option for active elderly hip fracture individuals. However, patients who are older, impaired or institutionalized benefit from HA.
Recurrence and metastasis remain the major obstacles to successful treatment of hepatocellular carcinoma (HCC). Chromatin remodeling factor ARID2 is commonly mutated in HCC, indicating its important ...role in cancer development. However, its role in HCC metastasis is largely elusive. In this study, we find that ARID2 expression is significantly decreased in metastatic HCC tissues, showing negative correlation with pathological grade, organ metastasis and positive association with survival of HCC patients. ARID2 inhibits migration and invasion of HCC cells in vitro and metastasis in vivo. Moreover, ARID2 knockout promotes pulmonary metastasis in different HCC mouse models. Mechanistic study reveals that ARID2 represses epithelial–mesenchymal transition (EMT) of HCC cells by recruiting DNMT1 to Snail promoter, which increases promoter methylation and inhibits Snail transcription. In addition, we discover that ARID2 mutants with disrupted C2H2 domain lose the metastasis suppressor function, exhibiting a positive association with HCC metastasis and poor prognosis. In conclusion, our study reveals the metastasis suppressor role as well as the underlying mechanism of ARID2 in HCC and provides a potential therapeutic target for ARID2-deficient HCC.
High-sensitivity C-reactive protein (hsCRP), a marker of inflammation, can promote atherosclerosis and predict cardiovascular events. However, no data are currently available about the combined ...effects of hsCRP and hypertension on cardiovascular risk. This study sought to elucidate this matter. A total of 7325 consecutive patients with angina-like chest pain undergoing coronary angiography were evaluated, and 4291 patients with stable, newly diagnosed coronary artery disease (CAD) were enrolled. They were subdivided into three groups according to baseline hsCRP levels (<1, 1-3, and >3 mg/L) and further stratified by hypertension status. The severity of CAD was assessed by the Gensini score and number of diseased vessels. All participants were followed for the occurrence of cardiovascular events. The coronary severity and cardiovascular outcomes were compared among these groups. We observed 530 (12.35%) incident cardiovascular events over 14,210 person-years. Elevated hsCRP was associated with more severe coronary lesions (p < 0.05) and an elevated but nonsignificant increased risk of cardiovascular events (p > 0.05). When hypertension was included as a stratifying factor, both patients with high hsCRP and normal blood pressure and hypertensive patients with any level of hsCRP had more severe coronary lesions compared with the reference group with low hsCRP and normotension. However, compared with the reference group, the cardiovascular event risk was only significantly elevated in patients with high hsCRP and hypertension (p < 0.05). The combination of elevated hsCRP and hypertension greatly increased the cardiovascular risk in patients with stable, newly diagnosed CAD, supporting that hsCRP could be treated as a marker for stratification in high-risk patients.
Identification of non-diabetic renal disease (NDRD) in patients with type 2 diabetes mellitus (T2DM) may help tailor treatment. Intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI) is a ...promising tool to evaluate renal function but its potential role in the clinical differentiation between diabetic nephropathy (DN) and NDRD remains unclear.
To investigate the added role of IVIM-DWI in the differential diagnosis between DN and NDRD in patients with T2DM.
Prospective.
Sixty-three patients with T2DM (ages: 22-69 years, 17 females) confirmed by renal biopsy divided into two subgroups (28 DN and 35 NDRD).
3 T/ T2 weighted imaging (T
WI), and intravoxel incoherent motion diffusion-weighted imaging (IVIM-DWI).
The parameters derived from IVIM-DWI (true diffusion coefficient D, pseudo-diffusion coefficient D*, and pseudo-diffusion fraction f) were calculated for the cortex and medulla, respectively. The clinical indexes related to renal function (eg cystatin C, etc.) and diabetes (eg diabetic retinopathy DR, fasting blood glucose, etc.) were measured and calculated within 1 week before MRI scanning. The clinical model based on clinical indexes and the IVIM-based model based on IVIM parameters and clinical indexes were established and evaluated, respectively.
Student's t-test; Mann-Whitney U test; Fisher's exact test; Chi-squared test; Intraclass correlation coefficient; Receiver operating characteristic analysis; Hosmer-Lemeshow test; DeLong's test. P < 0.05 was considered statistically significant.
The cortex D*, DR, and cystatin C values were identified as independent predictors of NDRD in multivariable analysis. The IVIM-based model, comprising DR, cystatin C, and cortex D*, significantly outperformed the clinical model containing only DR, and cystatin C (AUC = 0.934, 0.845, respectively).
The IVIM parameters, especially the renal cortex D* value, might serve as novel indicators in the differential diagnosis between DN and NDRD in patients with T2DM.
2 TECHNICAL EFFICACY: Stage 2.
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