Carboxypeptidase A6 suppresses the proliferation and invasion of colorectal cancer cells and is negatively regulated by miR-96-3p.

BACKGROUND: Colorectal cancer (CRC) is a common malignant tumor, and this study aims to explore the role and the regulatory mechanism of carboxypeptidase A6 (CPA6) in CRC cells. METHODS: Specific shRNA targeting CPA6 mRNA was transfected into NCM460 and HT29 cells to down-regulate CPA expression, and expression plasmid was transfected into HCT116 cells to exogenously overexpress CPA6. The dual luciferase assay was used to detect the direct binding of miR-96-3p to CPA6 3'UTR. Phosphorylation and activation of Akt were detected using Western blot. Cells were treated with miR-96-3p mimics, Akt inhibitor (MK-2206) or agonist (SC79) for rescue experiments. The cell functions were evaluated using CCK-8, clone formation, transwell, and Western blot assays. Xenograft tumor assay was also used to analyze the effect of altered CPA6 expression on tumor growth. RESULTS: Knockdown of CPA6 promoted the proliferation, clone formation, migration, and invasion of NCM460 and HT29 cells in vitro, and the tumor growth of nude mouse xenograft tumor in vivo. Moreover, over-expression of CPA6 significantly inhibited the malignant proliferation and invasion of HCT116 cells in vitro, and the tumor growth of xenograft tumor in vivo. Furthermore, miR-96-3p could directly regulate CPA6 expression by targeting its 3'UTR, and miR-96-3p mimics rescued the inhibitory effects of CPA6 overexpression on the malignant proliferation and invasion of CRC cells. Finally, CPA6 knockdown enhanced Akt/mTOR phosphorylation and activation, while CPA6 overexpression inhibited Akt/mTOR activation. The regulatory effect of CPA6 on Akt/mTOR signaling was naturally regulated by miR-96-3p. Akt inhibitor or agonist rescued the effects of CPA6 knockdown or overexpression on proliferation and EMT of colon cancer cells. CONCLUSION: CPA6 has a significant tumor suppressive effect on CRC by inhibiting the activation of Akt/mTOR signaling, and miR-96-3p negatively regulates the expression of CPA6.

A Synthetic Method for Site-Specific Functionalized Polypeptides: Metal-Free, Highly Active, and Selective at Room Temperature.

Functionalized polypeptides have attracted tremendous interest in recent years and found many stimulating applications owing to their tunable physicochemical characteristics including hydrophilicity and stimuli-responsive behavior. The development of new strategies to produce these polymers without metallic contaminants is crucial for their applications in high-value and sensitive domains, such as biomedical, microelectronic, food-packaging, and personal beauty care fields. Herein, a highly efficient strategy to access well-defined site-specific functionalized polypeptides is developed by combining Michael reaction with hydrogen-bonding organocatalytic ROP of NCA. A library of chain-end and chain-middle functionalized polypeptides (14 examples) with predesigned molecular weights and low polydispersities are readily prepared with this approach. Specifically, the whole synthetic process is metal-free, fulfilling high activity and selectivity at room temperature.

High-Temperature Superconductivity in Single-Unit-Cell FeSe Films on Anatase TiO_{2}(001).

We report on the observation of high-temperature (T_{c}) superconductivity and magnetic vortices in single-unit-cell FeSe films on anatase TiO_{2}(001) substrate by using scanning tunneling microscopy. A systematic study and engineering of interfacial properties has clarified the essential roles of substrate in realizing the high-T_{c} superconductivity, probably via interface-induced electron-phonon coupling enhancement and charge transfer. By visualizing and tuning the oxygen vacancies at the interface, we find their very limited effect on the superconductivity, which excludes interfacial oxygen vacancies as the primary source for charge transfer between the substrate and FeSe films. Our findings have placed severe constraints on any microscopic model for the high-T_{c} superconductivity in FeSe-related heterostructures.

MicroRNA-544a Regulates Migration and Invasion in Colorectal Cancer Cells via Regulation of Homeobox A10.

BACKGROUND/AIMS: MicroRNAs (miRNAs) are a group of small RNA molecules that post-transcriptionally regulate gene expression. Aberrant expression of miRNAs has been associated with tumorigenesis in various cancers. miR-544a is an understudied miRNA that has recently been implicated in regulating invasion in lung cancer. However, its role in regulating invasion and the underlying mechanism have not been investigated in colorectal cancer (CRC) cells. METHODS: Microarray analysis was performed in metastatic colorectal tumor samples and their matched normal tissues to identify differentially expressed miRNAs. Quantitative real-time PCR was used to detect miR-544a levels in tumor samples and CRC cell lines with varying metastatic properties. miR-544a mimic or inhibitor was transfected into SW480 and HCT116 cells, respectively, followed by wound healing and invasion assays. Western Blot and luciferase assay were performed to investigate the direct target of miR-544a. Xenograft mouse models was used to examine in vivo function of miR-544a. RESULTS: Our data showed that expression of miR-544a was significantly up-regulated in metastatic tumor samples and CRC cell lines. Inhibition of miR-544a reduced migration and invasion in HCT116 cells. Homeobox A10 (HOXA10) was the direct target of miR-544a which was required for the function of miR-544a in regulating invasiveness. miR-544a inhibitor and/or HOXA10 overexpression reduced lung metastases in HCT116 xenografts. CONCLUSIONS: Our study demonstrates that miR-544a regulates invasive and metastatic properties of CRC cells by modulating HOXA10 expression level both in vitro and in vivo. miR-544a may represent a new therapeutic target for the intervention of metastatic colorectal cancer.

Association of HMGB1 Gene Polymorphisms with Risk of Colorectal Cancer in a Chinese Population.

BACKGROUND Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. More advanced work is required in the detection of biomarkers for CRC susceptibility and prognosis. High-mobility group box-1 (HMGB1) is an angiogenesis-related gene reported to be associated with the development of CRC. The direct evidence of HMGB1 gene polymorphisms as biomarkers for CRC has not been reported previously. MATERIAL AND METHODS A total of 240 CRC patients and 480 healthy controls were periodically enrolled. DNA was extracted from blood specimens. The distributions of SNPs of HMGB1 were determined by using the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay. RESULTS In this case-control study, we observed a significant association between overall CRC risk and SNP rs2249825 (CG vs. CC and GG vs. CC). Participants carrying both rs2249825 CG (OR, 2.67; 95% CI, 1.89 to 3.78) and rs2249825 GG genotypes (OR, 2.32; 95% CI, 1.13 to 4.73) had a significantly increased risk of developing CRC compared to those carrying GG genotype. rs2249825 was associated with the risk of CRC in the dominant model but not in the recessive model. However, we found no significant differences in the rs1412125 or rs1045411 polymorphisms in the HMGB1. Advanced analyses showed that the number of rs2249825 G alleles showed a significant relationship with risk of CRC. CONCLUSIONS Our results show an association between HMGB1 rs2249825 SNP and CRC incidence in the Chinese Han population. However, population-based studies with more subjects and prognostic effects are needed to verify the association of HMGB1 SNPs with CRC susceptibility, severity, and long-term prognosis.

Mapping the Electronic Structure of Each Ingredient Oxide Layer of High-T\{c} Cuprate Superconductor Bi{2}Sr{2}CaCu{2}O{8+δ}.

Understanding the mechanism of high transition temperature (T{c}) superconductivity in cuprates has been hindered by the apparent complexity of their multilayered crystal structure. Using a cryogenic scanning tunneling microscopy (STM), we report on layer-by-layer probing of the electronic structures of all ingredient planes (BiO, SrO, CuO{2}) of Bi{2}Sr{2}CaCu_2}O{8+δ} superconductor prepared by argon-ion bombardment and annealing technique. We show that the well-known pseudogap (PG) feature observed by STM is inherently a property of the BiO planes and thus irrelevant directly to Cooper pairing. The SrO planes exhibit an unexpected van Hove singularity near the Fermi level, while the CuO{2} planes are exclusively characterized by a smaller gap inside the PG. The small gap becomes invisible near T{c}, which we identify as the superconducting gap. The above results constitute severe constraints on any microscopic model for high T{c} superconductivity in cuprates.

Knockdown of LRCH4 Remodels Tumor Microenvironment Through Inhibiting YAP and TGF-ß/Smad Signaling Pathway in Colorectal Cancer.

BACKGROUND: Colorectal cancer is one of the most common gastrointestinal malignancies worldwide. LRCH4 is the top 1 gene associated with an unfavorable prognosis in colorectal cancer. METHODS: Here, we reported that the knockdown of LRCH4 inhibited the proliferation, migration and invasion in HT29 cells. RESULTS: The activity of Yes-Associated Protein (YAP), a transcription factor in the Hppo-YAP signaling pathway, was significantly inhibited by LRCH4-siRNA. LRCH4 knockdown also reversed the EMT and regulated the expression of extracellular matrix (ECM) protein, Fibronectin and Collagen IV in HT29 cells. In addition, the TGF-ß/Smad signaling pathway, as the downstream pathway of Yap, was also inhibited by LRCH4 knockdown. CONCLUSION: Knockdown of LRCH4 involved in the regulation of ECM and EMT and inhibited YAP and the TGF-ß/Smad signaling pathway in colorectal cancer cells. Our study provided a mechanism of LRCH4 on colorectal cancer cells, and a new potential target for clinical tumor treatment.

Well-defined star (co)polypeptides via a fast, efficient, and metal-free strategy.

Polypeptides, especially star polypeptides, as a unique kind of biological macromolecules have broad applications in biomedical fields such as drug release, gene delivery, tissue engineering, and regenerative medicines due to their close structural similarity to naturally occurring peptides and proteins, biocompatibility, and amino acid functionality. However, the synthesis of star polypeptide mainly relies on the conventional primary amine-initiated ring-opening polymerization (ROP) of N-carboxyanhydrides (NCA) and suffers from low polymerization activity and limited controllability. This study proposes a fast, efficient and metal-free strategy to access star (co)polypeptides by combining the Michael reaction between acrylates and secondary aminoalcohols with the hydrogen-bonding organocatalytic ROP of NCA. This approach enables the preparation of a library of star (co)polypeptides with predesigned molecular weights, narrow molecular weight distributions, tunable arm number, and arm compositions. Importantly, this method exhibits high activity and selectivity at room temperature, making it both practical and versatile in synthesis applications.

Near-infrared light-induced homogeneous photoelectrochemical biosensor based on 3D walking nanomotor-assisted CRISPR/Cas12a for ultrasensitive microRNA-155 detection.

The dysregulation of microRNA (miRNA) expression levels is intricately linked to a myriad of human diseases, and the precise and delicate detection thereof holds paramount significance in the realm of clinical diagnosis and therapy. Herein, a near-infrared (NIR) light-mediated homogeneous photoelectrochemical (PEC) biosensor was constructed for miRNA-155 detection based on NaYF4: Yb, Tm@ZnIn2S4 (NYF@ ZIS) coupled with a three-dimensional (3D) walking nanomotor-assisted CRISPR/Cas12a strategy. The upconverted light emitted by the NYF in the visible and UV region upon NIR light excitation could be utilized to excite ZIS to produce a photocurrent response. The presence of target miRNA-155 initiated an amplification reaction within the 3D walking nanomotor, resulting in the production of multiple nucleic acid fragments. These fragments could activate the collateral cleavage capability of CRISPR/Cas12a, leading to the indiscriminate cleavage of single-stranded DNA (ssDNA) on ALP-ssDNA-modified magnetic beads and the subsequent liberation of alkaline phosphatase (ALP). The released ALP facilitated the catalysis of ascorbic acid 2-phosphate to generate ascorbic acid as the electron donor to capture the photogenerated holes on the NYF@ZIS surface, resulting in a positively correlated alteration in the photocurrent response. Under optimal conditions, the NIR light-initiated homogeneous PEC biosensor had the merits of good linear range (0.1 fM to 100 pM), an acceptable limit of detection (65.77 aM) for miRNA-155 detection. Considering the pronounced sensitivity, light stability, and low photodamage, this strategy presents a promising platform for detecting various other miRNA biomarkers in molecular diagnostic practice.

2D Z-scheme ZnIn2S4/g-C3N4 heterojunction based on photoelectrochemical immunosensor with enhanced carrier separation for sensitive detection of CEA.

Semiconducting materials based on photoelectrochemical (PEC) sensors have been widely utilized for detection. Meanwhile, the sensitivity of the PEC sensor was limited by low-efficiency carrier separation. Thus, a novel sandwich-type PEC bioimmunosensing based on 2D Z-scheme ZnIn2S4/g-C3N4 heterojunction as a photosensitive material and BiVO4 as a photoquencher was designed for the sensitive detection of carcinoembryonic antigen (CEA). Firstly, the 2D ZnIn2S4/g-C3N4 structure provided a multitude of activated sites which facilitated the loading of the capture antibody (Ab1). Secondly, the Z-scheme heterojunction had a high redox capacity while promoting the rapid separation and migration of photogenerated electron-hole pairs (e-/h+). Thus it was able to consume more electron donors to a certain extent, resulting in a higher initial photocurrent. In addition, BiVO4 with large spatial potential resistance was introduced for the first time to realize signal amplification. BiVO4 could not only compete with substrate materials for electron donors, but also effectively prevent electron donors from contacting the substrate, further reducing the photocurrent signal. Under optimized conditions, the sensor had a favorable detection range (0.0001-100 ng/mL) to CEA and a low detection limit of 0.03 pg/mL. With high specificity, excellent stability, and remarkable reproducibility, this sensor provided a new perspective for constructing accurate and convenient PEC immunosensor for bioanalysis and early disease diagnosisdisease diagnosis.

Self-powered photoelectrochemical immunosensing platform for sensitive CEA detection using dual-photoelectrode synergistic signal amplification.

Self-powered photoelectrochemical (PEC) sensing, as an emerging sensing mode, can effectively solve the problems such as weak anti-interference ability and poor signal response of individual photoanode or photocathode sensing. In this work, an ITO/Co-CuInS2 photocathode and ITO/WO3@CdS photoanode based self-powered cathodic PEC immunosensor was developed, which integrated dual-photoelectrode to synergistic amplify the signal for highly sensitive and specific detection of carcinoembryonic antigen (CEA). The self-powered PEC sensor could drive electrons transfer through the difference in Fermi levels between the two photoelectrodes without an external bias voltage. The photoanode was introduced to amplify the photoelectric signal, and the photocathode was only designed for the construction of sensing interfaces. The proposed sensor quantitatively determined the target CEA with the detection limit of 0.23 pg/mL and a linear correlation confine of 0.1 pg/mL ∼100 ng/mL. The constructed immunosensing platform exhibited high sensitivity, satisfactory stability and great biological detection selectivity, providing a feasible and effective strategy for the manufacture of new self-powered sensors in high-performance PEC bioanalytical applications.

Near-infrared light-induced photoelectrochemical biosensor based on plasmon-enhanced upconversion nanocomposites for microRNA-155 detection with cascade amplifications.

Herein, a novel near-infrared (NIR) light-driven photoelectrochemical (PEC) biosensor based on NaYF4:Yb3+, Er3+@Bi2MoO6@Bi (NYF@BMO@Bi) nanocomposites was elaborately developed to achieve highly sensitive detection of microRNA-155 (miRNA-155). To realize signal enhancement, the coupled plasmonic bismuth (Bi) nanoparticles were constructed as an energy relay to facilitate the transfer of energy from NaYF4:Yb3+, Er3+ to Bi2MoO6, ultimately enabling the efficient separation of electron-hole pairs of Bi2MoO6 under the irradiation of a 980 nm laser. For constructing biosensing system, the initial signal was firstly amplified after the addition of alkaline phosphatase (ALP) in conjunction with the biofunctionalized NYF@BMO@Bi nanocomposites, which could catalyze the conversion of ascorbic acid 2-phosphate into ascorbic acid, and then consumed the photoacoustic holes created on the surface of Bi2MoO6 for the enlarging photocurrent production. Upon addition of target miRNA-155, the cascade signal amplification process was triggered while the ALP-modified DNA sequence was replaced and then followed by the initiation of a simulated biocatalytic precipitation reaction to attenuate the photocurrent response. On account of the NIR-light-driven and cascade amplifications strategy, the as-constructed biosensor was successfully utilized for the accurate determination of miRNA-155 ranging from 1 fM to 0.1 µM with a detection limit of 0.32 fM. We believed that the proposed nanocomposites-based NIR-triggered PEC biosensor could provide a promising platform for effective monitoring other tumor biomarkers in clinical diagnostics.

Long-Term Exposure to Phenanthrene Induced Gene Expressions and Enzyme Activities of Cyprinus carpio below the Safe Concentration.

Phenanthrene (PHE) is a typical compound biomagnified in the food chain which endangers human health and generally accumulates from marine life. It has been listed as one of the 16 priority PAHs evaluated in toxicology. In order to evaluate the changes of CYP1A GST mRNA expression and EROD GST enzyme activity in carp exposed to lower than safe concentrations of PHE. Long-term exposure of carp to PHE at lower than safe concentrations for up to 25 days. The mRNA expression level and cytochrome P450 (CYP1A/EROD (7-Ethoxylesorufin O-deethylase)) and glutathione S-transferase (GST) activity were measured in carp liver and brain tissue. The results showed that PHE stress induced low-concentration induction and high-concentration inhibition of CYP1A expression and EROD enzyme activity in the liver and brain of carp. In both two organs, GST enzyme activity was also induced. However, the expression of GST mRNA was first induced and then inhibited, after the 15th day. These results indicate that long-term exposure to PHE at lower than safe concentrations still poses a potential threat to carp's oxidase system and gene expression.

A New Kind of Nonconventional Luminogen Based on Aliphatic Polyhydroxyurethane and Its Potential Application in Ink-Free Anticounterfeiting Printing.

Organic luminogens have extensive applications due to their unique photophysical properties. In recent years, nonconjugated organic luminogens, in contrast to traditional conjugated luminogens, have gained much attention because of their facile preparation, environmental friendliness, and biocompatibility. In this study, a new kind of nonconventional luminogen based on dynamic covalent cross-linked polyhydroxyurethane is reported for the first time. The new luminogen not only exhibits intrinsic strong fluorescent emission in the solid state but also possesses high mechanical properties along with good shape memory and self-healing properties. In addition, the new luminogens are synthesized from aliphatic polyfunctional cyclic carbonate and amines via a much more straightforward method, avoiding the use of toxic isocyanates. Investigations indicated that the intrinsic luminescence of the resultant luminogens was induced by the cross-linking of polymer chains and could be well tuned by controlling the degree of cross-linking. By taking advantage of the unique characteristics of the resultant polymer luminogens, we further developed a facile method, named "light-mediated ink-free screen printing", for anticounterfeiting paper fabrication. Different from traditional ink-based printing technology, the new method used UV-light instead of expensive security ink to encode anticounterfeiting information on natural cellulose paper. The anticounterfeiting information is stable under various wet conditions, showing promising applications in the fast-growing counterfeiting of pharmaceuticals, packaging, and the food industry.

Fast and selective organocatalytic ring-opening polymerization by fluorinated alcohol without a cocatalyst.

Organocatalysis is an important branch of catalysis for various organic transformations and materials preparation. Polymerizations promoted by organic catalysts can produce polymeric materials without any metallic residues, providing charming materials for high-value and sensitive domains such as biomedical applications, microelectronic devices and food packaging. Herein, we describe a fluorinated alcohol based catalytic system for polypeptide synthesis via catalytic ring-opening polymerization (ROP) of α-amino acid N-carboxyanhydride (NCA), fulfilling cocatalyst free, metal free, high rate and high selectivity. During polymerization, the fluorinated alcohol catalyst forms multiple dynamic hydrogen bonds with the initiator, monomer and propagating polymer chain. These cooperative hydrogen bonding interactions activate the NCA monomers and simultaneously protect the overactive initiator/propagating polymer chain-ends, which offers the whole polymerization with high activity and selectivity. Mechanistic studies indicate a monocomponent-multifunctional catalytic mode of fluorinated alcohol. This finding provides a metal free and fast approach to access well-defined polypeptides.

Long Non-Coding RNA TTN-AS1 Promotes the Proliferation and Invasion of Colorectal Cancer Cells by Activating miR-497-Mediated PI3K/Akt/mTOR Signaling.

INTRODUCTION: Long non-coding RNAs (lncRNAs) have obtained increasing attention due to their regulatory functions in many cancers. This work aimed to investigate the functional roles of lncRNA TTN-AS1 in colorectal cancer (CRC) and to explore the underlying mechanisms. METHODS: The expression profiles of TTN-AS1 and miR-497 in CRC tissues and cell lines were determined by RT-qPCR analysis. MTT assay, transwell assay, western blot analysis, and xenograft tumors in nude mice were employed to analyze the effects of TTN-AS1 on the proliferation, migration, invasion, EMT, and in vivo tumorigenesis of CRC cells. Bioinformatics analysis and dual-luciferase reporter assay determined the direct binding relation between TTN-AS1 and miR-497 in CRC. RESULTS: We observed a significant increase of TTN-AS1 expression level in CRC tissues and cell lines compared with normal counterparts. High expression of TTN-AS1 predicted a poor prognosis and was correlated with aggressive clinicopathological characteristics in CRC patients. Functionally, gain- and loss-of-function studies indicated that TTN-AS1 knockdown suppressed the proliferation, migration, invasion and epithelial-mesenchymal transition of CRC cells in vitro, whereas TTN-AS1 overexpression showed the complete opposite effects. Mechanistically, we found that TTN-AS1 could directly interact with miR-497, and co-transfection with miR-497 mimics blocked the activation of PI3K/Akt/mTOR signaling, and reversed the effects of TTN-AS1 overexpression in CRC cells. CONCLUSION: To conclude, our findings provide novel insight into CRC tumorigenesis and indicate that TTN-AS1 may serve as a potential therapeutic target for CRC treatment.

Vitrimer-Cellulose Paper Composites: A New Class of Strong, Smart, Green, and Sustainable Materials.

Vitrimer, the third category of polymer materials, combines the properties of traditional thermosets and thermoplastics and has gained much interest from industry since the first report in 2011. Currently, many researchers focus on the exploration of new chemistry for novel vitrimer synthesis but pay less attention to the fabrication of vitrimer composites based on known vitrimer systems. The latter can not only largely decrease the cost of vitrimers but also provide a facile way to increase the variety of vitrimer-based materials and extend the applications of vitrimers in different fields. In this study, we developed a new class of vitrimer composite using polycarbonate as a matrix and natural cellulose paper as the reinforcing framework for the first time. The resultant materials possess exceptional mechanical properties and great thermal/chemical stability, simultaneously exhibiting a series of smart properties, such as shape-memory, reshaping, self-healing, and reprocessing. Noteworthily, the two main components of the resultant materials, polycarbonate and natural cellulose, can be easily recycled under mild conditions; thus, these new vitrimer composites qualify as novel green and sustainable materials.

Occurrence and distribution of phthalate esters in freshwater aquaculture fish ponds in Pearl River Delta, China.

The concentrations, congener profiles and spatial distribution of 13 phthalate esters (PAEs) in the freshwater fish ponds in the Pearl River Delta (PRD) region were investigated in water and sediment samples collect from 22 sites during Jul. 2016-Sept. 2017. The di-2-ethylhexyl phthalate (DEHP) was the predominant compounds in both water and sediment samples, accounting for 70.1% and 66.1% of ∑PAEs, respectively. The DEHP concentrations in the water samples collected from the sites of Zhongshan (35.7 µg/L), Jingmen (17.3 µg/L) and Nanhai (14.2 µg/L) were higher than that collected from other sampling sites (p <0.05), and exceed the Chinese environmental quality standards for surface water (DEHP, 8.00 µg/L). The concentrations of ΣPAEs (mean and median were 11.8 mg/kg dw and 7.95 mg/kg dw) in sediment was higher than that in sediment of river and estuary in the PRD region (p <0.05). The median concentrations of DEHP and di-n-butyl phthalate (DBP) exceeded recommend environmental risk limit (ERL) that posed a potential risk to the aquaculture fish pond environment in the PRD.

The regulatory role of Fos related antigen­1 in inflammatory bowel disease.

The etiology of inflammatory bowel disease (IBD) remains unclear. The ratio of Fos related antigen­1 (Fra­1)­positive intestinal mucosa epithelial cells is significantly increased in active IBD. This study intends to explore the regulatory role of Fra­1 in IBD. The Fra­1 eukaryotic expression vector was constructed and stably transfected to establish the Fra­1 overexpression HCT­116 (116­Fra­1) intestinal epithelial cell line. The impact of Fra­1 overexpression on intestinal mucosal epithelial cell damage repair function was tested using a scratch assay. The role of Fra­1 overexpression on intestinal mucosal epithelial cell proliferation was evaluated using a Cell Counting Kit-8 assay. Apoptosis related proteins, B­cell lymphoma 2 (Bcl­2), c­Myc, Survivin and Bcl­extra large (Bcl­xL), expression levels were detected by western blotting. Fra­1 suppressed intestinal mucosal epithelial cell damage repair and proliferation. Fra­1 inhibited the protein levels of Bcl­2, c­Myc, Survivin, and Bcl­xL. Fra­1 overexpression in intestinal mucosal epithelial cells may restrain damage repair after intestinal mucosal injury in IBD remittent period through weakening the protective effect of intestinal mucosa, thus increasing the risk of recurrence. Therefore, suppressing Fra­1 expression in intestinal mucosal epithelial cells may contribute to IBD remittent maintenance and recurrence delay.

Determination of Anthracene in Water Samples by ELISA Using AhR Binding.

The degradation of polycyclic aromatic hydrocarbons (PAHs) can generate AhR-binding compounds, exhibiting genotoxicity and carcinogenicity. In this investigation, aryl hydrocarbon receptor (AhR) from carp and anthracene (Ant) were coupled as antigen to establish an indirect competition ELISA (ic-ELISA) with an AhR-Ant antibody. A standard curve was determined for the ic-ELISA concerning detection range and limit. Also, the specificity, stability and the recovery of the ic-ELISA were checked. Results indicate that the ratio of antibody to antigen titer is 1:64000. The resulting standard curve is Y = 21.326 × X + 1.8213. The detection range lies within 10 - 1000 ng mL-1 and the limiting concentration is 2.43 ng mL-1. The cross reaction ratio (CR) between Ant and naphthalene (Nap), Ant and phenanthrene (Phe) or Ant and fluoranthene (Flu) were 5.7, 19.1 and less than 0.1%, respectively. The range of the coefficient of variance (C.V) amounts was from 4.2 up to 9.5% and the recovery range was from 90 to 115%. These results show that the AhR-Ant ic-ELISA is sensitive, and can be used as a technical support to quantify Ant in the environment.