Skin injury in adult mammals brings about a series of events and inflammation in the wounded area is initiated first and provides lots of inflammatory factors, which is critical for the final scar formation. While the postinjured skin of fetus and nude mice heals scarlessly owing to the absence of inflammation or immunodeficient, we designed a feasible acid-responsive ibuprofen-loaded poly(L-lactide) (PLLA) fibrous scaffolds via doping sodium bicarbonate to prevent excessive inflammation and achieve scarless healing finally. The morphological results of in vivo experiments revealed that animals treated with acid-responsive ibuprofen-loaded PLLA fibrous scaffolds exhibited alleviative inflammation, accelerated healing process, and regulated collagen deposition via interference in the collagen distribution, the ?-smooth muscle actin (?-SMA), and the basic fibroblast growth factor (bFGF) expression. The lower ratios of collagen I/collagen III and TGF-?1/TGF-?3 and higher ratio of matrix metalloproteinase-1 (MMP-1)/tissue inhibitor of metalloproteinase-1 (TIMP-1) in acid-responsive ibuprofen-loaded PLLA fibrous scaffolds group were confirmed by real-time qPCR as well. These results suggest that inhibiting the excessive inflammation will result in regular collagen distribution and appropriate ratio between the factors, which promote or suppress the scar formation, then decrease the scar area, and finally achieve the scarless healing.
The present study aimed to quantitatively determine the aberrant methylation signal of the adenomatous polyposis coli (APC) gene in hepatocellular carcinoma (HCC), and to evaluate whether hypermethylation of the APC promoter could be a prognostic biomarker for HCC. Taqman probe-based quantitative methylation-specific polymerase chain reaction was performed to identify the APC promoter methylation levels in 57 HCC and corresponding non-tumorous liver tissues. In the present study, the methylation level of the APC promoter was upregulated by 4.51-fold in the HCC tissues compared with the non-cancerous tissues (P=0.0003). With regard to the clinicopathological data, the methylation level of the APC promoter in the HCC samples was higher in the patients with larger tumors when the cut-off was set at 4 cm (P=0.0008), and in the older patients when the cut-off was set at 60 years old (P=0.0438). However, the methylation status in the HCC samples appeared not to affect the overall patient survival rate (P=0.1684). The findings of the present study showed that APC promoter hypermethylation accumulates during the development of HCC, but that it may not be a promising prognostic biomarker for HCC.
The fast evaluation of the discrete Fourier transform of an image at non-uniform sampling locations is key to efficient iterative non-Cartesian MRI reconstruction algorithms. Current non-uniform fast Fourier transform (NUFFT) approximations rely on the interpolation of oversampled uniform Fourier samples. The main challenge is high memory demand due to oversampling, especially when multidimensional datasets are involved. The main focus of this work is to design an NUFFT algorithm with minimal memory demands. Specifically, we introduce an analytical expression for the expected mean square error in the NUFFT approximation based on our earlier work. We then introduce an iterative algorithm to design the interpolator and scale factors. Experimental comparisons show that the proposed optimized NUFFT scheme provides considerably lower approximation errors than the previous designs  that rely on worst case error metrics. The improved approximations are also seen to considerably reduce the errors and artifacts in non-Cartesian MRI reconstruction.
A novel high-throughput purification method for a monocot mannose-binding lectin, Pinellia ternata agglutinin (PTA), from tubers of P. ternata was established by mannose-Sephrose 4B affinity chromatography. The total protein was extracted from tubers of P. ternata using phosphate buffered saline (PBS) buffer. The extracted total protein was precipitated completely at 65% ammonium sulfate saturation and dissolved in different concentrations of NaCl solution to activate its binding affinity toward the column. PTA was bound to the affinity column by loading of the total protein into the column and elution using PBS buffer. The maximum purification yield (35.5mg/g) was obtained when PTA was treated with 25% (w/v) NaCl solution, and the purity of PTA analyzed by SDS-PAGE was ?97%. The agglutination property of purified PTA was confirmed by mouse erythrocytes, which indicates its biological function. Nuclear staining assay and DNA fragmentation demonstrated that PTA could induce apoptosis of Bel-7404 cells, which further demonstrates its biological and pharmacological activities. Induction of apoptosis in the human tumor Bel-7404 cell line by PTA indicates its possible use in cancer therapy. The present investigation reports a significantly improved isolation method to obtain highly purified mannose-binding plant lectin proteins. The proposed method has great potential for industrial application because of its advantages, which include rapid isolation, high purity, high yield, low cost, and minimal requirement of chemical materials.
microRNAs (miRNAs) are involved in the pathogenesis of diverse human cancers through its target genes, including papillary thyroid cancer (PTC). However, there are few studies regarding associations between clinicopathological features of PTC with the expression of specific miRNAs and its potential target genes. In the present study, analysis of miRNA was integrated with mRNA expression profiles in aggressive PTC. miRNA and gene expression arrays were used to identify a subset of differentially expressed miRNAs and mRNAs between aggressive and non-aggressive PTCs. These miRNAs and mRNAs were further validated by qPCR in a cohort of 20 PTCs with extrathyroidal invasion and/or distant metastases, and 20 PTCs with no extrathyroidal invasion. The target of these miRNAs was determined by luciferase reporter and bioinformatic analysis. The miRNA arrays identified 14 upregulated miRNAs and 10 downregulated miRNAs in aggressive compared with non-aggressive PTCs. Significant miRNA deregulation was confirmed in the validation cohort, with upregulation of miR-146b-5p and miR-221/222 and downregulation of miR-16 and miR-613 in aggressive PTCs. The gene arrays identified 2000 differentially expressed genes, in which TIMP3, ZNFR3, FN1 and ITGA2 were observed to be target genes inversely correlated with miR-221/222, miR-146b-5p, miR-613 and miR-16, respectively. The results of the present study indicated the potential importance of miR-221/222, miR-146b-5p, miR-16 and miR-613 in determining the aggressive properties of PTC by targeting TIMP3, ZNFR3, FN1 and ITGA2, respectively. Additional studies should be conducted to confirm the results.
Papillary thyroid cancer (PTC) is the most common form of well-differentiated thyroid carcinoma. Although the incidence of PTC has been on the increase in China, the precise causes of the disease remain unknown and a prognostic index has yet to be identified. In the present study, quantitative real?time PCR and western blot analysis were used to detect the mRNA and protein expression levels, respectively, of forkhead box E1 (FOXE1) in 30 PTC tissue specimens matched with adjacent non-tumor thyroid tissues. FOXE1 expression in 81 PTC paraffin-embedded blocks was also evaluated retrospectively by immunohistochemistry (IHC) and its correlation with the clinicopathological parameters of PTC patients was analyzed. Compared with adjacent non-tumor tissues, the mRNA expression levels of the FOXE1 gene in tumor tissues were significantly higher in 65% of PTC tissue specimens. The results from western blotting and IHC analysis were consistent with FOXE1 mRNA expression. Furthermore, the expression of FOXE1 was significantly correlated with the extra-capsular invasion of tumor cells, lymph node metastasis and tumor stage (P=0.048, P=0.036 and P=0.009, respectively). Thus, our results demonstrated that the expression of FOXE1 was significantly increased in PTC tumor tissues and correlated with clinical prognosis. Therefore, FOXE1 is a potential biomarker for PTC prognosis as well as a new therapeutic target.
Pinellia ternata agglutinin (PTA) from the tubers of P. ternata is a two-domain monocot mannose-binding lectin. Pta-n encoding N-terminus domain of PTA (PTA-N) was fused with Escherichia coli alkaline phosphatase signal peptide (APSP) gene by polymerase chain reaction (PCR) for secretion expression. The fused nucleotide sequence apsp-pta-n was inserted into pET-28a prokaryotic expression vector by restriction enzyme digest sites (Nco I and Xho I), and then overexpressed in E. coli BL21(DE3) cells by isopropyl ?-d-1-thiogalactopyranoside (IPTG) induction. Expressed APSP targeted the recombinant protein APSP-PTA-N into the periplasmic space, and then APSP was recognized and automatically cleaved by the membrane-bound signal peptidase. Ni-NTA chromatography was used for the purification and about 20 mg/L purified PTA-N was obtained. The minimum agglutination concentration of PTA-N determined by mice erythrocytes was 6.33 ± 0.47 ?g/ml. The carbohydrate inhibition assay was carried out to determine the carbohydrate-binding property indicating PTA-N bound to specific sugars. The in vitro anti-proliferative activity towards human tumor cell lines and anti-fungal activity against Gibberella saubinetii were also demonstrated. Nuclear staining assay was performed to demonstrate PTA-N induced cell apoptosis. The results showed that PTA-N had significant biological functions, similar to native PTA. This strategy was the first time used to express plant mannose-binding lectin proteins and the product induced human tumor cell apoptosis, suggesting its potential application in biomedicine research.
Laparoscopic Nissen fundoplication (LNF) is the standard procedure for surgical management of gastro-oesophageal reflux disease (GORD). Laparoscopic Toupet fundoplication (LTF) is reported to be as effective as LNF but to be associated with a lower incidence of post-operative dysphagia. This meta-analysis was performed to compare the two techniques with respect to reflux control and associated complications, particularly dysphagia.
We introduce a novel noniterative algorithm for the fast and accurate reconstruction of nonuniformly sampled MRI data. The proposed scheme derives the reconstructed image as the nonuniform inverse Fourier transform of a compensated dataset. We derive each sample in the compensated dataset as a weighted linear combination of a few measured k-space samples. The specific k-space samples and the weights involved in the linear combination are derived such that the reconstruction error is minimized. The computational complexity of the proposed scheme is comparable to that of gridding. At the same time, it provides significantly improved accuracy and is considerably more robust to noise and undersampling. The advantages of the proposed scheme makes it ideally suited for the fast reconstruction of large multidimensional datasets, which routinely arise in applications such as f-MRI and MR spectroscopy. The comparisons with state-of-the-art algorithms on numerical phantoms and MRI data clearly demonstrate the performance improvement.
Toll-like receptor4 (TLR4) plays an important role in the induction and regulation of the innate or adaptive immune responses. Thus, the genetic variation in TLR4 gene may influence the development of autoimmune diseases such as rheumatoid arthritis (RA). Several studies have investigated the roles of genetic polymorphisms of TLR4 gene in RA, but most of these studies were restricted to two cosegregating functional missense polymorphisms Asp299Gly and Thr399Ile. To determine whether non-missense genetic polymorphisms located in regulatory region of TLR4 are related to RA in a Chinese Han population, four single nucleotide polymorphisms (SNPs) situated on 3 untranslating region (UTR) and 5 UTR were genotyped in 213 RA patients and 247 unrelated ethnically matched controls using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and direct sequencing techniques. Significant genetic associations were observed with the 3 UTR SNP rs41426344 and rs7873784. The minor allele C and homozygotic variant genotype CC of rs41426344 and minor allele C of rs7873784 were identified to be risk factors for the development of RA in Chinese Han people. Furthermore, by comparing the variation allele frequencies to other populations, prevalent genetic ethnic specificity was observed in all the four SNPs. Our study suggested that the effect of non-missense polymorphisms located in regulatory region would not be neglected in disease association analysis.
p53 tumor suppressor gene Arg72Pro polymorphism has been associated with bladder cancer. However, results were inconsistent. We performed this meta-analysis to estimate the association between p53 Arg72Pro polymorphism and bladder cancer.
We introduce a unifying energy minimization framework for nonlocal regularization of inverse problems. In contrast to the weighted sum of square differences between image pixels used by current schemes, the proposed functional is an unweighted sum of inter-patch distances. We use robust distance metrics that promote the averaging of similar patches, while discouraging the averaging of dissimilar patches. We show that the first iteration of a majorize-minimize algorithm to minimize the proposed cost function is similar to current nonlocal methods. The reformulation thus provides a theoretical justification for the heuristic approach of iterating nonlocal schemes, which re-estimate the weights from the current image estimate. Thanks to the reformulation, we now understand that the widely reported alias amplification associated with iterative nonlocal methods are caused by the convergence to local minimum of the nonconvex penalty. We introduce an efficient continuation strategy to overcome this problem. The similarity of the proposed criterion to widely used nonquadratic penalties (e.g., total variation and lp semi-norms) opens the door to the adaptation of fast algorithms developed in the context of compressive sensing; we introduce several novel algorithms to solve the proposed nonlocal optimization problem. Thanks to the unifying framework, these fast algorithms are readily applicable for a large class of distance metrics.
The use of CD133 as a marker for identifying and isolating colon cancer stem cells (CSCs) is controversial. In particular, it is not clear whether the pattern of expression is influenced by the tumor microenvironment. We analysed the expression of CD133 in the SW620 colon cancer cell line, which produces subpopulations with different levels of CD133. CD133(neg) and CD133(hi) SW620 cells were isolated and compared in hanging drop spheroid cultures. The expression of CD133 was examined under different culture conditions and following nutrient withdrawal. The results showed that SW620 cells express the CD133 antigen at variable levels. CD133(neg) and CD133(hi) SW620 cells can be separated by fluorescence-activated cell sorting and, when grown as adherent cells in liquid culture, are serially passaged without changes to the CD133 levels. However, the CD133 antigen is re-expressed when the CD133(neg) cells are deprived of nutrients or grown as non-adherent cells in spheroid cultures. Our observations indicate that the presence of the CD133 antigen does not identify a stem cell population of SW620 colorectal cancer cells. The expression of CD133 is dynamic and is modulated during certain cell culture conditions, suggesting that changes in expression may be mediated by the energy supply available to the cells.
Papillary thyroid carcinoma (PTC) is the most common carcinoma of the thyroid gland and has a relatively favorable prognosis. However, it is important to identify PTC characteristics that indicate a high risk for recurrence and metastasis. Recent data indicate that the amyloid precursor protein (APP) is involved in cell adhesion, motility and proliferation. At present, the expression levels of APP and their prognostic significance in PTC have not been studied. In the present study, the APP gene expression in 10 PTCs and their adjacent thyroid tissue samples was analyzed using quantitative real time-PCR. Tissue array blocks were obtained from 90 PTC cases. Tumor regions and their adjacent non-tumor regions were analyzed in tissue array blocks and immunohistochemistry was conducted using sectioned slides. Semi-quantitative scores were correlated with the clinicopathological factors of 90 PTC patients. APP-specific mRNA was increased 50-fold in PTC samples compared with their adjacent thyroid tissue. The immunohistochemistry results showed APP expression levels to be significantly increased in PTC samples compared with their adjacent non-tumor thyroid tissue (p<0.001). High APP scores were significantly correlated with large tumor size, extracapsular invasion and lymph node metastasis. The amyloid precursor protein was overexpressed in PTC and a high APP expression was associated with high malignant potential. Therefore, APP may serve as a prognostic marker and potential novel therapeutic target in PTC.
MicroRNAs (miRNAs) are regulators of numerous cellular events; accumulating evidence indicates that miRNAs play a key role in a wide range of biological functions, such as cellular proliferation, differentiation, and apoptosis in cancer. Down-regulated expression of miR-145 has been reported in colon cancer tissues and cell lines. The molecular mechanisms underlying miR-145 and the regulation of colon carcinogenesis remain unclear. In this study, we investigated the levels of miR-145 in human colon cancer cells using qRT-PCR and found markedly decreased levels compared to normal epithelial cells. We identified PAK4 as a novel target of miR-145 using informatics screening. Additionally, we demonstrated that miR-145 targets a putative binding site in the 3UTR of PAK4 and that its abundance is inversely associated with miR-145 expression in colon cancer cells; we confirmed this relationship using the luciferase reporter assay. Furthermore, restoration of miR-145 by mimics in SW620 cells significantly attenuated cell growth in vitro, in accordance with the inhibitory effects induced by siRNA mediated knockdown of PAK4. Taken together, these findings demonstrate that miR-145 downregulates P-ERK expression by targeting PAK4 and leads to inhibition of tumor growth.
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