Nicotinamide Riboside Regulates Chemotaxis to Decrease Inflammation and Ameliorate Functional Recovery Following Spinal Cord Injury in Mice.

Changes in intracellular nicotinamide adenine dinucleotide (NAD+) levels have been observed in various disease states. A decrease in NAD+ levels has been noted following spinal cord injury (SCI). Nicotinamide riboside (NR) serves as the precursor of NAD+. Previous research has demonstrated the anti-inflammatory and apoptosis-reducing effects of NR supplements. However, it remains unclear whether NR exerts a similar role in mice after SCI. The objective of this study was to investigate the impact of NR on these changes in a mouse model of SCI. Four groups were considered: (1) non-SCI without NR (Sham), (2) non-SCI with NR (Sham +NR), (3) SCI without NR (SCI), and (4) SCI with NR (SCI + NR). Female C57BL/6J mice aged 6-8 weeks were intraperitoneally administered with 500 mg/kg/day NR for a duration of one week. The supplementation of NR resulted in a significant elevation of NAD+ levels in the spinal cord tissue of mice after SCI. In comparison to the SCI group, NR supplementation exhibited regulatory effects on the chemotaxis/recruitment of leukocytes, leading to reduced levels of inflammatory factors such as IL-1ß, TNF-α, and IL-22 in the injured area. Moreover, NR supplementation notably enhanced the survival of neurons and synapses within the injured area, ultimately resulting in improved motor functions after SCI. Therefore, our research findings demonstrated that NR supplementation had inhibitory effects on leukocyte chemotaxis, anti-inflammatory effects, and could significantly improve the immune micro-environment after SCI, thereby promoting neuronal survival and ultimately enhancing the recovery of motor functions after SCI. NR supplementation showed promise as a potential clinical treatment strategy for SCI.

Remote limb ischemic preconditioning alleviated spinal cord injury through inhibiting proinflammatory immune response and promoting the survival of spinal neurons.

STUDY DESIGN: Experimental animal study. OBJECTIVES: To investigate the protective effect of remote limb ischemia preconditioning (RLPreC) on traumatic spinal cord injury (SCI) and explore the underlying biological mechanisms using RNA sequencing. SETTING: China Rehabilitation Science Institute; Beijing; China. METHODS: spinal cord injury was induced in mice using a force of 0.7 N. RLPreC treatment was administered. Motor function, pain behavior, and gene expression were assessed. RESULTS: RLPreC treatment significantly improved motor function and reduced pain-like behavior in SCI mice. RNA-Seq analysis identified 5247 differentially expressed genes (DEGs). GO analysis revealed enrichment of immune response, inflammatory signaling, and synaptic transmission pathways among these DEGs. KEGG analysis indicated suppression of inflammation and promotion of synapse-related pathways. CONCLUSIONS: RLPreC is a promising therapeutic strategy for improving motor function and alleviating pain after traumatic SCI. RNA-Seq analysis provides insights into potential therapeutic targets and warrants further investigation.

Biological Response to Recombinant Human Bone Morphogenetic Protein-2 on Bone-Implant Osseointegration in Ovariectomized Experimental Design.

OBJECTIVE: The aim of this study is to investigate the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) on bone-implant osteointegration in osteoporotic rats. MATERIALS AND METHODS: Thirty-six female Wistar rats were randomly divided into 3 groups: sham-operation (SHAM), ovariectomized (OVX), and ovariectomized with rhBMP-2 (OVX + rhBMP-2). The bone density of right tibia was observed with x-ray and the serum alkaline phosphatase (ALP) activity was measured preovariectomy and postovariectomy using an ALP-kit. In OVX + rhBMP-2 group, rhBMP-2 was embedded in the peri-implant area, while SHAM and OVX groups did not contain rhBMP-2. Four and eight weeks after implantation, the rats were killed and the right tibia with implants was taken by x-ray. Histologic changes were investigated by hematoxylin and eosin staining, scanning electron microscope (SEM), and energy dispersive spectrometer examinations. RESULTS: The serum ALP level was significantly higher in ovariectomized rats compared with that before ovariectomy (P < 0.05), while no difference was found in SHAM rats. At 12 weeks after ovariectomy, radiographic and histologic findings showed significant osteoporotic changes in proximal tibial metaphyses of OVX rats, including reduced cortical bone density and enlargement of bone marrow cavity compared with SHAM ones. The results of implantation verified new bone formation around implants in OVX + rhBMP-2 and SHAM groups, indicating favorable bone healing and osseointegration. No bone resorption was found in OVX + rhBMP-2 group, while some soft tissue was observed in bone-implant interface in SHAM group. In OVX group, there was no effective bone-implant osseointegration and mature bone formed around implants, and some implants were even lost due to chronic inflammation. The percentage of calcium and phosphorous atoms was significantly higher and the percentage of sulfur element was significantly lower in peri-implant area in OVX + rhBMP-2 and SHAM groups than that in OVX group. CONCLUSION: rhBMP-2 could enhance the osseous healing and restore bone-implant osseointegration in osteoporotic rats.

Urban Land Use and Land Cover Classification Using Novel Deep Learning Models Based on High Spatial Resolution Satellite Imagery.

Urban land cover and land use mapping plays an important role in urban planning and management. In this paper, novel multi-scale deep learning models, namely ASPP-Unet and ResASPP-Unet are proposed for urban land cover classification based on very high resolution (VHR) satellite imagery. The proposed ASPP-Unet model consists of a contracting path which extracts the high-level features, and an expansive path, which up-samples the features to create a high-resolution output. The atrous spatial pyramid pooling (ASPP) technique is utilized in the bottom layer in order to incorporate multi-scale deep features into a discriminative feature. The ResASPP-Unet model further improves the architecture by replacing each layer with residual unit. The models were trained and tested based on WorldView-2 (WV2) and WorldView-3 (WV3) imageries over the city of Beijing. Model parameters including layer depth and the number of initial feature maps (IFMs) as well as the input image bands were evaluated in terms of their impact on the model performances. It is shown that the ResASPP-Unet model with 11 layers and 64 IFMs based on 8-band WV2 imagery produced the highest classification accuracy (87.1% for WV2 imagery and 84.0% for WV3 imagery). The ASPP-Unet model with the same parameter setting produced slightly lower accuracy, with overall accuracy of 85.2% for WV2 imagery and 83.2% for WV3 imagery. Overall, the proposed models outperformed the state-of-the-art models, e.g., U-Net, convolutional neural network (CNN) and Support Vector Machine (SVM) model over both WV2 and WV3 images, and yielded robust and efficient urban land cover classification results.

Circulating Endothelial Progenitor Cells were Increased in Patients with Thin-Cap Fibroatheroma.

BACKGROUND: We used virtual histology-intravascular ultrasound (VH-IVUS) to evaluate the relationship between circulating endothelial progenitor cells (EPC) and thin-cap fibroatheroma (TCFA). METHODS: This study included 52 patients with unstable angina who underwent coronary angiography and IVUS examination. Patients were divided into a TCFA group (n = 21) or a non-TCFA group (n = 31) based on VH-IVUS performance. Before angiography, peripheral blood levels of EPC were measured by flow cytometry. TCFA was defined as a necrotic core (NC) ≥ 10% of the plaque area without overlying fibrous tissue in the presence of ≥ 40% plaque burden. RESULTS: Levels of circulating EPCs were 72.45 ± 31.73 (count/105) in the TCFA group and 23.93 ± 11.87 (count/ 105) (p < 0.01). Mean levels of CRP were 0.38 ± 0.21 mg/L in the TCFA group and 0.23 ± 0.17 mg/L (p < 0.01). Levels of EPCs correlated positively with necrotic core volume(r = 0.421, p = 0.005), CRP (r = 0.405, p = 0.011) and negatively with fibrous tissue volume(r = -0.411, p = 0.009). In multivariate logistic regression analysis, level of EPC (OR: 1.815, 95% CI: 1.12 - 2.798, p = 0.016), plaque burden (OR: 1.26, 95% CI: 1.07 - 1.86, p = 0.027), and CRP (OR; 1.14, 95% CI: 0.74 - 1.56, p = 0.029) were independent predictors of TCFA. CONCLUSIONS: Circulating EPCs were increased in patients with TCFA, level of EPCs could predict the presence of TCFA.

Media Use, Cognitive Performance, and Life Satisfaction of the Chinese Elderly.

Media use and aging is an important interdisciplinary topic pertaining to communication, gerontology, and psychology, among others. Integrating research on media-induced recovery and life satisfaction, the present study examined media uses and effects in the context of health and aging. Specifically, data from a large random sample were analyzed to investigate the relationships between media use, cognitive performance, and life satisfaction among the Chinese elderly. Results, in general, lent support to the slightly modified structural model. Specifically, media-induced recovery outcomes can be categorized into proximate (e.g., cognitive performance), intermediate (e.g., health satisfaction), and distal levels (e.g., life satisfaction). Also, situational factors (e.g., disease history) had statistically significant effects on media-induced recovery outcomes. Theoretical and practical implications of the research findings were discussed, and future research directions were suggested.

Cereal-derived arabinoxylans as biological response modifiers: extraction, molecular features, and immune-stimulating properties.

Arabinoxylans are of significant importance to human health due to their potential to modulate both the adaptive and innate immune systems. Arabinoxylans of various structures and sources have been shown to affect different immune cells to augment a wide range of immune responses in vitro and in vivo in animals and humans. This review article discusses current research on the immune-enhancing activities of arabinoxylans and other cereal polysaccharides in relation to their structural heterogeneity. There are inconsistencies in the literature regarding the relationships between the immunomodulatory effects and the structure and source of arabinoxylans. Possible mechanisms underlying these relationships which might explain the effects of such bioactive polysaccharides are proposed.

Enhancement of Stopping Power Ratio (SPR) Estimation Accuracy through Image-Domain Dual-Energy Computer Tomography for Pencil Beam Scanning System: A Simulation Study.

Our study aims to quantify the impact of spectral separation on achieved theoretical prediction accuracy of proton-stopping power when the volume discrepancy between calibration phantom and scanned object is observed. Such discrepancy can be commonly seen in our CSI pediatric patients. One of the representative image-domain DECT models is employed on a virtual phantom to derive electron density and effective atomic number for a total of 34 ICRU standard human tissues. The spectral pairs used in this study are 90 kVp/140 kVp, without and with 0.1 mm to 0.5 mm additional tin filter. The two DECT images are reconstructed via a conventional filtered back projection algorithm (FBP) on simulated noiseless projection data. The best-predicted accuracy occurs at a spectral pair of 90 kVp/140 kVp with a 0.3 mm tin filter, and the root-mean-squared average error is 0.12% for tissue substitutes. The results reveal that the selected image-domain model is sensitive to spectral pair deviation when there is a discrepancy between calibration and scanning conditions. This study suggests that an optimization process may be needed for clinically available DECT scanners to yield the best proton-stopping power estimation.

Role of microbiota-gut-brain axis in natural aging-related alterations in behavior.

Introduction: Aging is a complex, time-dependent biological process that involves a decline of overall function. Over the past decade, the field of intestinal microbiota associated with aging has received considerable attention. However, there is limited information surrounding microbiota-gut-brain axis (MGBA) to further reveal the mechanism of aging. Methods: In this study, locomotory function and sensory function were evaluated through a series of behavioral tests.Metabolic profiling were determined by using indirect calorimetry.16s rRNA sequence and targeted metabolomics analyses were performed to investigate alterations in the gut microbiota and fecal short-chain fatty acids (SCFAs). The serum cytokines were detected by a multiplex cytokine assay.The expression of proinflammatory factors were detected by western blotting. Results: Decreased locomotor activity, decreased pain sensitivity, and reduced respiratory metabolic profiling were observed in aged mice. High-throughput sequencing revealed that the levels of genus Lactobacillus and Dubosiella were reduced, and the levels of genus Alistipes and Bacteroides were increased in aged mice. Certain bacterial genus were directly associated with the decline of physiological behaviors in aged mice. Furthermore, the amount of fecal SCFAs in aged mice was decreased, accompanied by an upregulation in the circulating pro-inflammatory cytokines and increased expression of inflammatory factors in the brain. Discussion: Aging-induced microbial dysbiosis was closely related with the overall decline in behavior, which may attribute to the changes in metabolic products, e.g., SCFAs, caused by an alteration in the gut microbiota, leading to inflammaging and contributing to neurological deficits. Investigating the MGBA might provide a novel viewpoint to exploring the pathogenesis of aging and expanding appropriate therapeutic targets.

ECM-mimetic glucomannan hydrogel promotes pressure ulcer healing by scavenging ROS, promoting angiogenesis and regulating macrophages.

Pressure ulcers (PUs) have emerged as a significant burden on both individuals and society. Effective treatment of PUs is a significant clinical challenge due to the compromised tissue microenvironment characterized by extracellular matrix (ECM) depletion, increased levels of reactive oxygen species (ROS), excessive inflammation and impaired angiogenesis. To this end, we have developed a glucomannan hydrogel (GM-Pgel) that mimics the skin's extracellular matrix to accelerate wound healing by regulating chronic inflammation in the PUs. This hydrogel not only faithfully replicates the components and nanofibrous architecture of ECM-like glycoproteins but also exhibits remarkable capabilities in enhancing neovascularization, scavenging ROS, and promoting macrophage polarization toward the M2 phenotype. In summary, this ECM-mimetic multifunctional hydrogel emerges as a promising dressing with diverse functionalities, capable of reshaping the compromised tissue environment without the need for additional drugs, exogenous cytokines, or cells. This presents a compelling and effective strategy for the repair and regeneration of chronic cutaneous wounds.

Long non-coding RNA LINC01296 promotes progression of oral squamous cell carcinoma through activating the MAPK/ERK signaling pathway via the miR-485-5p/PAK4 axis.

Introduction: Long intergenic non-protein-coding RNA 1296 (LINC01296), a newly identified lncRNA, can function as an oncogenic driver to promote the development of multiple carcinomas. However, the effect of LINC01296 on oral squamous cell carcinoma (OSCC) is still unclear. Material and methods: We determined the expression and role of LINC01296 in OSCC tissues and cell lines. The cell viability, migration and invasion were determined by MTT, wound healing assay and transwell assay, respectively. Flow cytometry was used for detecting cell cycle and apoptosis. The interaction and association between LINC01296, microRNA-485-5p (miR-485-5p) and p21 (RAC1) activated kinase 4 (PAK4) were analyzed by RNA immunoprecipitation (RIP) and luciferase reporter assays. The xenograft mouse model was established to detect the effect of LINC01296 on OSCC tumor growth. Results: Our study showed that LINC01296 was over-expressed in OSCC tissues and cell lines. The level of LINC01296 was positively correlated with the patient's tumor node metastasis (TNM) stage and nodal invasion. Knockdown of LINC01296 effectively inhibits cell viability, migration and invasion but promotes cell apoptosis in vitro. The in vivo experiment showed that LINC01296 knockdown inhibited OSCC tumor growth. The following analysis indicated that LINC01296 acted as a ceRNA for miR-485-5p, and PAK4 was identified as a direct target of miR-485-5p. Furthermore, we found that the effects of LINC01296 on OSCC progression were through regulating the expression of PAK4/p-MEK/p-ERK via sponging miR-485-5p. Conclusions: LINC01296 promote the cell cycle, proliferation, migration and invasion, and inhibit apoptosis of OSCC cells through activating the MAPK/ERK signaling pathway via sponging miR-485-5p to regulate PAK4 expression. These results suggested that the LINC01296/miR-485-5p/PAK4 axis was closely associated with OSCC progression. Our study provides a new insight into the molecular pathogenesis of OSCC, and may supply novel biomarkers for diagnosis and therapy of OSCC.

MMP1 Overexpression Promotes Cancer Progression and Associates with Poor Outcome in Head and Neck Carcinoma.

Matrix metalloproteinase-1 (MMP1) has been reported to play key roles in a variety of cancers by degrading the extracellular matrix. However, its carcinogenic roles have not been shown yet in head and neck squamous cell carcinoma (HNSCC). This study aimed to elucidate its expression pattern and functional roles as well as clinical significance in HNSCC. The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), and immunohistochemistry (IHC) were utilized to determine the MMP1 expression pattern and the associations between its expression and patients' outcome in HNSCC. Mice tongue squamous cell carcinoma model induced by 4-nitroquinoline 1-oxide (4NQO) and siRNA-mediated cellular assay in vitro were utilized to evaluate the oncogenic role of MMP1. The biological functions and cancer-related pathways involved in MMP1-related genes were found through bioinformatics analysis. Both mRNA and protein abundance of MMP1 were highly increased in HNSCC as compared to its non-tumor counterparts. MMP1 overexpression positively correlated with advanced tumor size, cervical node metastasis, and advanced pathological grade and lower patients' survival. In the 4NQO-induced animal model, MMP1 expression increased along with the progression of the disease. In HNSCC cells, siRNA-mediated knockdown of MMP1 significantly inhibited cell proliferation, migration, and invasion and activated apoptosis and epithelia-mesenchymal transition (EMT). GSEA, GO, and KEGG analyses showed that MMP1 expression was significantly related to cancer-related pathways and cancer-related functions. Together, our results demonstrated MMP1 serves as a novel prognostic biomarker and putative oncogene in HNSCC.

Structure of Machupo virus polymerase in complex with matrix protein Z.

The Arenaviridae family includes several viruses that cause severe human hemorrhagic fevers with high mortality, with no effective countermeasures currently available. The arenavirus multi-domain L protein is involved in viral transcription and replication and represents a promising target for antiviral drugs. The arenavirus matrix protein Z is a small multi-functional protein that inhibits the activities of the L protein. Here we report the structure of Machupo virus L protein in complex with Z determined by cryo-electron microscopy. The Z protein acts as a staple and binds the L protein with 1:1 stoichiometry at the intersection between the PA-C-like region, RNA-dependent RNA polymerase and PB2-N-like region. Binding of the Z protein may lock the multiple domains of L into a fixed arrangement leading to loss of catalytic activity. These results further our understanding of the inhibitory mechanism of arenavirus replication machinery and provide a novel perspective to develop antiviral drugs.

Characterization and clinical evaluation of microsatellite instability and loss of heterozygosity in tumor-related genes in gastric cancer.

Microsatellite instability (MSI) detection is widely used in the diagnosis and prognosis evaluation of colorectal cancer. However, for gastric cancer (GC), there is no standard panel of microsatellites (MSs) used in clinical guidance. The present study aimed to identify useful predictors of the clinical features and for the prognosis of GC, based on an investigation of MSI and loss of heterozygosity (LOH) in tumor-related genes. First, from 20 tumor-related genes which were proven to be important to the development of GC, 91 MSs were identified, and PCR amplification, short tandem repeat scanning analysis and TA clone sequencing were used to analyze MSI and LOH in the first set of 90 GC samples. Subsequently, the same method was used to detect the MSI/LOH of the optimized loci in the second set of 136 GC samples. MSI/LOH in the mismatch repair genes was highly consistent with that in oncogenes and tumor suppressor genes, respectively. The length of the core sequence was a main factor for the MSI/LOH rate. The MSI of 12 single loci was significantly associated with lymph node metastasis. The MSI in TP53-1 and the LOH in MGMT-10 were significantly associated with early stages of tumor infiltration depth. The LOH in MGMT-10, PTN-2 and MCC-17 was significantly associated with TNM stage. The LOH in TP53-1 and ERBB2-12 was associated with adenocarcinoma. The MSI/LOH in 6 single loci of 5 tumor-related genes was associated with poor prognosis of GC. The present study demonstrated that the MSI/LOH of loci in tumor-associated genes was associated with 4 clinicopathological characteristics and outcomes of GC. These results may provide potential specific biomarkers for the clinical prediction and treatment of GC.

Design and system evaluation of a dual-panel portable PET (DP-PET).

BACKGROUND: Integrated whole-body PET/MR technology continues to mature and is now extensively used in clinical settings. However, due to the special design architecture, integrated whole-body PET/MR comes with a few inherent limitations. Firstly, whole-body PET/MR lacks sensitivity and resolution for focused organs. Secondly, broader clinical access of integrated PET/MR has been significantly restricted due to its prohibitively high cost. The MR-compatible PET insert is an independent and removable PET scanner which can be placed within an MRI bore. However, the mobility and configurability of all existing MR-compatible PET insert prototypes remain limited. METHODS: An MR-compatible portable PET insert prototype, dual-panel portable PET (DP-PET), has been developed for simultaneous PET/MR imaging. Using SiPM, digital readout electronics, novel carbon fiber shielding, phase-change cooling, and MRI compatible battery power, DP-PET was designed to achieve high-sensitivity and high-resolution with compatibility with a clinical 3-T MRI scanner. A GPU-based reconstruction method with resolution modeling (RM) has been developed for the DP-PET reconstruction. We evaluated the system performance on PET resolution, sensitivity, image quality, and the PET/MR interference. RESULTS: The initial results reveal that the DP-PET prototype worked as expected in the MRI bore and caused minimal compromise to the MRI image quality. The PET performance was measured to show a spatial resolution ≤ 2.5 mm (parallel to the detector panels), maximum sensitivity = 3.6% at the center of FOV, and energy resolution = 12.43%. MR pulsing introduces less than 2% variation to the PET performance measurement results. CONCLUSIONS: We developed a MR-compatible PET insert prototype and performed several studies to begin to characterize the performance of the proposed DP-PET. The results showed that the proposed DP-PET performed well in the MRI bore and would cause little influence on the MRI images. The Derenzo phantom test showed that the proposed reconstruction method could obtain high-quality images using DP-PET.

Impact of bowtie filter and detector collimation on multislice CT scatter profiles: A simulation study.

PURPOSE: To investigate via Monte Carlo simulations, the impact of scan subject size, antiscatter grid (ASG), collimator size, and bowtie filter on the distribution of scatter radiation in a typical realistically modeled third generation 16 slice diagnostic computed tomography (CT) scanner. METHODS: Full radiation transport was simulated with Geant4 in a realistic CT scanner geometric model, including the imaging phantom, bowtie filter (BTF), collimators and detector assembly, except for the ASGs. An analytical method was employed to quantify the probable transmission through the ASG of each photon intersecting the detector array. Normalized scatter profiles (NSP) and scatter-to-primary-ratio (SPR) profiles were simulated for 90 and 140 kVp beams for different size phantoms and slice thicknesses. The impact of CT scatter on the reconstructed attenuation coefficient factor was also studied as were the modulating effects of phantom- and patient-tissue heterogeneities on scatter profiles. A method to characterize the relative spatial frequency content of sinogram signals was developed to assess the latter. RESULTS: For the 21.4-cm diameter phantom, NSP and SPR increase linearly with collimator opening for both tube potentials, with the 90 kVp scan exhibiting slightly larger NSP and SPR. The BTF modestly modulates scatter under the phantom center, reducing the prominent off-axis lobes by factors of 1.1-1.3. The ASG reduces scatter on the central axis NSP threefold, and reduces scatter at the detectors outside the phantom shadow by factors of 25 to 500. For the phantoms with diameters of 27 and 32 cm, the scatter increases roughly three- and fourfold, respectively, demonstrating that scatter monotonically increases with phantom size, despite deployment of the ASG and BTF. In the absence of a scan subject, the ASG reduces the signal profile arising photons scattered by the BTF. Without ASG, the in-air scatter profile is relatively flat compared to the scatter profile when the ASG is present. For both 90 and 140 kVp photon spectra, the calculated attenuation coefficient decreases linearly with increasing collimation size. For both homogeneous and heterogeneous objects, NSPs are dominated by low spatial frequency content compared to the primary signal. However, the SPR, which quantifies the local magnitude of nonlinear detector response and is dominated by the high frequency content of the primary profile, can contribute strongly to high-spatial frequency streaking artifacts near high-density structures in reconstructed image artifacts. CONCLUSION: Public-domain Monte Carlo codes, Geant-4 in particular, is a feasible method for characterizing CT detector response to scattered- and off-focal radiation. Our study demonstrates that the ASG substantially reduces the scatter radiation and reshapes scatter-radiation profiles and affects the accuracy with which the detector array can measure narrow-beam attenuation due its inability to distinguish between true uncollided primary and narrow-angle coherently scattered photons. Hence, incorporating the impact of detector array collimation into the forward-projection signal formation models used by iterative reconstruction algorithms is necessary to use CT for accurately characterizing material properties. While tissue heterogeneities exercise a modest influence on local NPS shape and magnitude, they do not add significant high spatial frequency content.

Characterization and clinical evaluation of microsatellite instability and loss of heterozygosity within tumor-related genes in colorectal cancer.

BACKGROUND: Microsatellite instability (MSI) is a biomarker for better outcomes in colorectal cancer (CRC). However, this conclusion is controversial. In addition, MSs can be a useful marker for loss of heterozygosity (LOH) of genes, but this finding has not been well studied. Here, we aimed to clarify the predictive value of MSI/LOH within tumor-related genes in CRC. METHODS: We detected MSI/LOH of MSs in tumor-related genes and the Bethesda (B5) panel by STR scanning and cloning/sequencing. We further analyzed the relationship between MSI/LOH status and clinical features or outcomes by Pearson's Chi-square test, Fisher's exact test and the Kaplan-Meier method. RESULTS: The findings indicated that the MSI rates of B5 loci were all higher than those of loci in tumor-related genes. Interestingly, MSI/LOH of 2 loci in the B5 panel and 12 loci in tumor-related genes were associated with poorer outcomes, while MSI/LOH of the B5 panel failed to predict outcomes in CRC. MSI of BAT25, MSI/LOH of BAT26 and MSI of the B5 panel showed closer relationships with mucinous carcinoma. In addition, LOH-H of the B5 panel was associated with increased lymphatic metastasis. CONCLUSIONS: In summary, MSI/LOH of certain loci or the whole panel of B5 is related to clinical features, and several loci within tumor-related genes showed prognostic value in the outcomes of CRC.

Radial forearm free flap for reconstruction of a large defect after radical ablation of carcinoma of the tongue and floor of the mouth: some new modifications.

BACKGROUND: A modified radial forearm free flap was designed to rehabilitate function and to reduce the complications at both donor and recipient sites. METHODS: Between 2003 and 2007, 15 patients with infiltrating squamous cell carcinoma (T(3)-T(4)) of the tongue and/or floor of the mouth underwent hemiglossectomy and resection of the floor of the mouth with microvascular reconstruction using a modified radial forearm flap. The mean size of the forearm flap was 7.5 x 14 cm, and the de-epithelialized area was 7 x 6 cm, requiring no skin graft from the abdomen. Speech intelligibility tests were administered to test postoperative speech and the functional oral intake scale was applied to assess the postoperative swallowing function, and patients reconstructed with pectoralis major myocutaneous flap were used for comparison. RESULTS: All the flaps were successfully transferred. No obvious complications were found in either the oral-maxillofacial or forearm region. The speech intelligibility was better in the modified flap group (p < 0.01). An acceptable swallowing function was also achieved, although the difference was not significant (p > 0.05). CONCLUSIONS: The modified flap used for reconstructing large defects of the tongue and floor of the mouth might be a valid substitute for pectoralis major myocutaneous flap to improve the outcome in individuals with significant oral carcinoma.

Hyperinsulinemia impairs functions of circulating endothelial progenitor cells.

AIMS: Circulating endothelial progenitor cells (EPCs) play a key role in maintaining endothelial function. Dysfunction of EPCs is associated with the cardiovascular complication of diabetes. The purpose of this study is to investigate the direct effects of hyperinsulinemia on EPCs and the underlying mechanisms. METHODS: EPCs isolated from healthy adults were cultured with various concentrations of insulin (control group, without insulin; physiological insulin group, 10 nM insulin and hyperinsulinemia group, 100 nM insulin) with or without phosphatidylinositol-3-kinase (PI3-K) inhibitor (LY294002, 5 µM), endothelial nitric oxide synthase (eNOS) inhibitor (L-NG-nitro-arginine methyl ester (L-NAME), 100 µM), sodium nitroprusside (SNP, 25 µM), p38 mitogen-activated protein kinase(MAPK) inhibitor (SB203580, 5 µM) or extracellular signal-regulated kinases (ERK) 1/2 inhibitor (PD98059, 10 µM). Proliferation, tube formation, and apoptosis of EPCs were determined. Expressions of eNOS, PI3-K, protein kinase B (Akt), p38 MAPK, and ERK 1/2 were assessed. RESULTS: Hyperinsulinemia caused a significant decrease in proliferation and tube formation abilities than control group. Hyperinsulinemia increased apoptosis rate of EPCs than control group. Furthermore, hyperinsulinemia downregulated phosphorylation of eNOS, PI3-K and Akt, and upregulated phosphorylation of p38 MAPK and ERK. SNP could restore impaired tube formation induced by hyperinsulinemia. P38 MAPK inhibitor but not ERK inhibitor could decrease apoptosis induced by hyperinsulinemia. CONCLUSION: Hyperinsulinemia impaired EPCs' tube formation ability by downregulation of PI-3K/Akt/eNOS pathway. Hyperinsulinemia induced apoptosis of EPCs via upregulation of p38 MAPK.