Potential prognostic and predictive value of UBE2N, IMPDH1, DYNC1LI1 and HRASLS2 in colorectal cancer stool specimens.

Colorectal cancer (CRC) is the most common gastrointestinal malignancy worldwide. The poor specificity and sensitivity of the fecal occult blood test has prompted the development of CRC-related genetic markers for CRC screening and treatment. Gene expression profiles in stool specimens are effective, sensitive and clinically applicable. Herein, a novel advantage of using cells shed from the colon is presented for cost-effective CRC screening. Molecular panels were generated through a series of leave-one-out cross-validation and discriminant analyses. A logistic regression model following reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry was used to validate a specific panel for CRC prediction. The panel, consisting of ubiquitin-conjugating enzyme E2 N (UBE2N), inosine monophosphate dehydrogenase 1 (IMPDH1), dynein cytoplasmic 1 light intermediate chain 1 (DYNC1LI1) and phospholipase A and acyltransferase 2 (HRASLS2), accurately recognized patients with CRC and could thus be further investigated as a potential prognostic and predictive biomarker for CRC. UBE2N, IMPDH1 and DYNC1LI1 expression levels were upregulated and HRASLS2 expression was downregulated in CRC tissues. The predictive power of the panel was 96.6% [95% confidence interval (CI), 88.1-99.6%] sensitivity and 89.7% (95% CI, 72.6-97.8%) specificity at a predicted cut-off value at 0.540, suggesting that this four-gene panel testing of stool specimens can faithfully mirror the state of the colon. On the whole, the present study demonstrates that screening for CRC or cancer detection in stool specimens collected non-invasively does not require the inclusion of an excessive number of genes, and colonic defects can be identified via the detection of an aberrant protein in the mucosa or submucosa.

Downregulated Calcium-Binding Protein S100A16 and HSP27 in Placenta-Derived Multipotent Cells Induce Functional Astrocyte Differentiation.

Little is known about genes that induce stem cells differentiation into astrocytes. We previously described that heat shock protein 27 (HSP27) downregulation is directly related to neural differentiation under chemical induction in placenta-derived multipotent stem cells (PDMCs). Using this neural differentiation cell model, we cross-compared transcriptomic and proteomic data and selected 26 candidate genes with the same expression trends in both omics analyses. Those genes were further compared with a transcriptomic database derived from Alzheimer's disease (AD). Eighteen out of 26 candidates showed opposite expression trends between our data and the AD database. The mRNA and protein expression levels of those candidates showed downregulation of HSP27, S100 calcium-binding protein A16 (S100A16) and two other genes in our neural differentiation cell model. Silencing these four genes with various combinations showed that co-silencing HSP27 and S100A16 has stronger effects than other combinations for astrocyte differentiation. The induced astrocyte showed typical astrocytic star-shape and developed with ramified, stringy and filamentous processes as well as differentiated endfoot structures. Also, some of them connected with each other and formed continuous network. Immunofluorescence quantification of various neural markers indicated that HSP27 and S100A16 downregulation mainly drive PDMCs differentiation into astrocytes. Immunofluorescence and confocal microscopic images showed the classical star-like shape morphology and co-expression of crucial astrocyte markers in induced astrocytes, while electrophysiology and Ca2+ influx examination further confirmed their functional characteristics. In conclusion, co-silencing of S100A16 and HSP27 without chemical induction leads to PDMCs differentiation into functional astrocytes.

Promotion of the Differentiation of Dental Pulp Stem Cells into Oligodendrocytes by Knockdown of Heat Shock Protein 27.

Stem cell-based therapy has been evaluated in many different clinical trials for various diseases. This capability was applied in various neurodegenerative diseases, such as multiple sclerosis, which is characterized by demyelination, axonal injury, and neuronal loss. Dental pulp stem cells (DPSCs) are mesenchymal stem cells from the oral cavity that have been studied with potential application for the regeneration of different tissues. Heat shock protein 27 (HSP27) regulates neurogenesis in the process of neural differentiation of placenta multipotent stem cells. Here, we hypothesize that HSP27 expression is also critical for the neural differentiation of DPSCs. An evaluation of the possible role of HSP27 in the differentiation of DPSCs was performed using gene knockdown and neural immunofluorescent staining. We found that HSP27 played a role in the differentiation of DPSCs and that knockdown of HSP27 in DPSCs rendered cells to oligodendrocyte progenitors; i.e., small hairpin specific for HSP27 DPSCs exhibited NG2-positive immunoreactivity and gave rise to oligodendrocytes or type-2 astrocytes. This neural differentiation of DPSCs may have clinical significance in the treatment of patients with neurodegenerative diseases. In conclusion, our data provide an example of the oligodendrocyte differentiation of a DPSC model, which may be applied in human regenerative medicine.

The Occurrence of Pain-Induced Depression Is Different between Rat Models of Inflammatory and Neuropathic Pain.

Various pain conditions may be associated with depressed mood. However, the effect of inflammatory or neuropathic pain on depression-like behavior and its associated time frame has not been well established in rat models. This frontward study investigated the differences in pain behavior, depression-like behavior, and serotonin transporter (SERT) distribution in the brain between rats subjected to spared nerve injury (SNI)-induced neuropathic pain or complete Freund's adjuvant (CFA)-induced inflammatory pain. A dynamic plantar aesthesiometer and an acetone spray test were used to evaluate mechanical and cold allodynia responses, and depression-like behavior was examined using a forced swimming test and sucrose preference test. We also investigated SERT expression by using positron emission tomography. We found that the inflammation-induced pain was less severe than neuropathic pain from days 3 to 28 after induced pain; however, the CFA-injected rats exhibited more noticeable depression-like behavior and had significantly reduced SERT expression in the brain regions (thalamus and striatum) at an early stage (on days 14, 21, and 28 in two groups of CFA-injected rats versus day 28 in SNI rats). We speculated that not only the pain response after initial injury but also the subsequent neuroinflammation may have been the crucial factors influencing depression-like behavior in rats.

Comparative functional genomic analysis of Alzheimer's affected and naturally aging brains.

BACKGROUND: Alzheimer's disease (AD) is a prevalent progressive neurodegenerative human disease whose cause remains unclear. Numerous initially highly hopeful anti-AD drugs based on the amyloid-ß (Aß) hypothesis of AD have failed recent late-phase tests. Natural aging (AG) is a high-risk factor for AD. Here, we aim to gain insights in AD that may lead to its novel therapeutic treatment through conducting meta-analyses of gene expression microarray data from AG and AD-affected brain. METHODS: Five sets of gene expression microarray data from different regions of AD (hereafter, ALZ when referring to data)-affected brain, and one set from AG, were analyzed by means of the application of the methods of differentially expressed genes and differentially co-expressed gene pairs for the identification of putatively disrupted biological pathways and associated abnormal molecular contents. RESULTS: Brain-region specificity among ALZ cases and AG-ALZ differences in gene expression and in KEGG pathway disruption were identified. Strong heterogeneity in AD signatures among the five brain regions was observed: HC/PC/SFG showed clear and pronounced AD signatures, MTG moderately so, and EC showed essentially none. There were stark differences between ALZ and AG. OXPHOS and Proteasome were the most disrupted pathways in HC/PC/SFG, while AG showed no OXPHOS disruption and relatively weak Proteasome disruption in AG. Metabolic related pathways including TCA cycle and Pyruvate metabolism were disrupted in ALZ but not in AG. Three pathogenic infection related pathways were disrupted in ALZ. Many cancer and signaling related pathways were shown to be disrupted AG but far less so in ALZ, and not at all in HC. We identified 54 "ALZ-only" differentially expressed genes, all down-regulated and which, when used to augment the gene list of the KEGG AD pathway, made it significantly more AD-specific.

Electrical Field-Assisted Gene Delivery from Polyelectrolyte Multilayers.

To sustain gene delivery and elongate transgene expression, plasmid DNA and cationic nonviral vectors can be deposited through layer-by-layer (LbL) assembly to form polyelectrolyte multilayers (PEMs). Although these macromolecules can be released for transfection purposes, their entanglement only allows partial delivery. Therefore, how to efficiently deliver immobilized genes from PEMs remains a challenge. In this study, we attempt to facilitate their delivery through the pretreatment of the external electrical field. Multilayers of polyethylenimine (PEI) and DNA were deposited onto conductive polypyrrole (PPy), which were placed in an aqueous environment to examine their release after electric field pretreatment. Only the electric field perpendicular to the substrate with constant voltage efficiently promoted the release of PEI and DNA from PEMs, and the higher potential resulted in the more releases which were enhanced with treatment time. The roughness of PEMs also increased after electric field treatment because the electrical field not only caused electrophoresis of polyelectrolytes and but also allowed electrochemical reaction on the PPy electrode. Finally, the released DNA and PEI were used for transfection. Polyplexes were successfully formed after electric field treatment, and the transfection efficiency was also improved, suggesting that this electric field pretreatment effectively assists gene delivery from PEMs and should be beneficial to regenerative medicine application.

Enhanced Perioperative Safety and Comfort During Airway-Related Surgeries and Procedures With Dexmedetomidine-A Brief Review and Clinical Practice Experience.

Dexmedetomidine, an α-2 adrenergic receptor agonist, provides analgesia, sedation, anxiolysis, sympatholysis and anesthetic-sparing effect, without inducing significant respiratory depression. Due to these properties, its clinical use is no longer limited to serving as a sedative agent in the intensive care unit. Proper airway management and the avoidance of cardiac and respiratory complications are common goals of everyday anesthesia practice. Ensuring airway safety is pivotal during the anesthesia stages of induction, maintenance and recovery. In this review, we focus on the advantages of dexmedetomidine in awake fiberoptic intubation (AFOI), diagnostic examinations and surgeries of patients with obstructed airways, and reducing emergence delirium effectively without causing further adverse events. With increasing implementation in different anesthetic scenarios, dexmedetomidine provides a favorable option to enhance patient safety and comfort.

A Randomized Controlled Study of the Use of Video Double-Lumen Endobronchial Tubes Versus Double-Lumen Endobronchial Tubes in Thoracic Surgery.

OBJECTIVE: To compare the incidence of fiberoptic bronchoscope (FOB) use (1) during verification of initial placement and (2) for reconfirmation of correct placement following repositioning, when either a double-lumen tube (DLT) or video double-lumen tube (VDLT) was used for lung isolation during thoracic surgery. DESIGN: A randomized controlled study. SETTING: Single-center university teaching hospital. PARTICIPANTS: The study comprised 80 patients who were 18 years or older requiring lung isolation for surgery. INTERVENTIONS: After institutional review board approval, patients were randomized prior to surgery to either DLT or VDLT usage. Attending anesthesiologists placed the Mallinckrodt DLT or Vivasight (ET View Ltd, Misgav, Israel) VDLT with conventional laryngoscopy or video laryngoscopy then verified correct tube position through the view provided with either VDLT external monitor or FOB. MEASUREMENTS AND MAIN RESULTS: Data collected included: sex, body mass index, successful intubation and endobronchial placement, intubation time, confirmation time of tube position, FOB use, quality of view, dislodgement of tube, and ability to forewarn dislodgement of endobronchial cuff and complications. FOB use for verification of final position of the tube (VDLT 13.2% [5/38] v DLT 100% [42/42], p < 0.0001), need for FOB to correct the dislodgement (VDLT 7.7% [1/13] v DLT 100% [14/14], p < 0.0001), dislodgement during positioning (VDLT 61.5% [8/13] v DLT 64.3% [9/14], p = ns), dislodgement during surgery (VDLT 38.5% [5/13] v DLT 21.4% [3/14], p = ns), and ability to forewarn dislodgement of endobronchial cuff (VDLT 18.4% [7/38] v DLT 4.8% [2/42], p = 0.078). CONCLUSION: This study demonstrated a reduction of 86.8% in FOB use, which was a similar reduction found in other published studies.

MiR-29a and miR-652 Attenuate Liver Fibrosis by Inhibiting the Differentiation of CD4+ T Cells.

BACKGROUND: Liver fibrosis is the response of liver diseases that puzzles patients. MiRNAs were involved in the regulating processes of liver fibrosis. This study aims to investigate the effects of ARRB1 mediated by miR-29a and miR-652 on liver fibrosis and its possible mechanism. METHODS: Liver fibrosis of mice was induced by intraperitoneal injection of CCl4. Liver function was observed by the levels of alanine transaminase (ALT) and aspartate transaminase (AST). Flow cytometry was used to detect the percent of T helper17 (Th17). ELISA (Enzyme linked immunoassay) was used to detect the levels of Interleukin-17 (IL-17) and Interleukin-22 (IL-22). Real-time PCR was used to detect the expression of IL-17A, IL-22, miR-29a, miR-652 and ß-Arrestin 1 Gene (ARRB1). Western blot was used to detect the protein expression of ARRB1. RESULTS: CCl4 supplementation significantly increased the level of ALT and AST, the percent of Th17, the level of IL-17A, IL-22, miR-29a and miR-652, but decreased ARRB1. Overexpression of miR-29a/miR-652 prominently decreased Th17, IL-17A, IL-22 and ARRB1 in the normal CD4+ T cells. Both miR-29a and miR-652 targeted ARRB1 to regulate its expression. The effects of miR-29a/miR-652 overexpression on CD4+ T cells were reversed by ARRB1 overexpression. In vivo experiments demonstrated the protective role of miR-29a/miR-652 overexpression on liver fibrosis. CONCLUSION: ARRB1 mediated by miR-29a and miR-652 probably involved in the CD4+ T cells differentiation in patients with liver fibrosis, and functioned as a biomarker of fibrosis liver.Key words: liver fibrosis, miR-29a, miR-652, ARRB1, CD4+ T cells.

Cystatin C in cerebrospinal fluid is upregulated in elderly patients with chronic osteoarthritis pain and modulated through matrix metalloproteinase 9-specific pathway.

OBJECTIVES: Although high abundant cystatin c (CysC) in cerebrospinal fluid (CSF) is well known, its ambiguous role associated with pain still remains unclear. This study evaluated the effects of intrathecal CysC content from chronic pain caused by osteoarthritis (OA) and the novel relationship with matrix metalloproteinases 2 and 9 (MMP2 and MMP9) in CSF. METHODS: Samples of CSF were obtained from 8 elderly patients (65 y and above) with OA with lower limb pain for at least 6 months (OA group) and 8 sex-matched and age-matched relatively healthy elderly individuals without any pain problems (control group). The intrathecal CysC, MMP2, and MMP9 were examined by Western blotting. The analysis of CysC cleavage under different conditions was performed through silver staining and using mass-spectroscopy (SELDI-TOF) on 2 groups. RESULTS: Expression of full-length CysC and pro-MMP2 proteins showed statistically significant upregulation (P=0.0004 vs. 0.03), and expression of MMP9 protein showed downregulation (P=0.007) in the OA group. Both MMP9 and MMP2 initiated the mechanism for full-length CysC cleavage only in the presence of CSF. However, MMP9 showed greater ability than MMP2 for CysC cleavage in control and OA groups in sliver staining. Incubation of CSF with the MMP9 inhibitor led to the suppression of CysC cleavage in SELDI-TOF. DISCUSSION: These findings provide the first in vivo evidence on a relationship between CysC and gelatinases (MMP2 and MMP9), and could facilitate further investigation of novel interactions among these proteins within the proteomics field, especially protein-protein interactions involved in pain.

[Efficacy and safety of vitamin D in the treatment of idiopathic oligoasthenozoospermia].

OBJECTIVE: To explore the efficacy and safety of vitamin D (VD) in the treatment of idiopathic oligoasthenozoospermia. METHODS: This study included 86 infertile men with idiopathic oligoasthenozoospermia, who were randomized to a VD and a control group of equal number, the former given oral VD 200 IU/d and calcium 600 mg/d,qd, while the latter administered oral vitamin E 100 mg and vitamin C 100 mg, tid. After 3 months of medication, we compared the semen parameters, adverse reactions, and pregnancy rate between the two groups. RESULTS: After medication, the count of progressively motile sperm per ejaculate was increased from (9.82 ± 3.72) x 10(6) to (21.47 ± 6.52) x 10(6) ( P < 0.05) and the proportion of progressively motile sperm from (18.41 ± 9.82)% to (28.27 ± 4.47)% (P < 0.05) in the VD group. In comparison, the count of progressively motile sperm per ejaculate was elevated from (9.51 ± 6.31) x 10(6) to (12.36 ± 4.43) x 10(6) (P > 0.05) and the proportion of progressively motile sperm from (17.79 ± 5.25)% to (21.35 ± 2.41)% (P > 0.05) in the control group. Pregnancy was achieved in 7 cases (16.3%) in the VD group, but only lease (2.3%) in the control (P < 0.05). No adverse reactions were observed in either of the groups. CONCLUSION: Vitamin D, as a safe option for the treatment of idiopathic oligoasthenozoospermia, can effectively improve the semen quality, especially the progressive sperm motility of the patient.

A fast universal immobilization of immunoglobulin G at 4 °C for the development of array-based immunoassays.

To maintain the antibody activity and enhance performance of array-based immunoassays, protein G was used to allow a shorter duration of immunoglobulin G immobilization at 4 °C, with the antibody placed in the appropriate orientation. The multiplexed detection of six pain-related message molecules (PRMMs) was used as examples for the development of array-based immunoassays: substance P, calcitonin gene-related peptide, nerve growth factor, brain-derived neurotrophic factor, tumor necrosis factor-α, and ß-endorphin. Protein G- and non-protein G-coated slides were tested. Compared to non-protein G immunoassays, protein G shortened the antibody immobilization time at 4 °C from overnight to 2 hours. Only protein G-facilitated immunoassays succeeded in simultaneously detecting all six PRMMs with high specificity. Dose-response curves showed that the limits of detection of the protein G-multiplexed immunoassays for the PRMMs was approximately 164, 167, 120, 60, 80, and 92 pg/ml, respectively. Thus, protein G effectively shortens the duration of antibody immobilization at 4 °C, allowing the use of sensitive array-based immunoassays for the simultaneous detection of PRMMs.

Ultra-low dose naloxone restores the antinocicepitve effect of morphine in PTX-treated rats: association of IL-10 upregulation in the spinal cord.

AIMS: Ultra-low dose naloxone has been shown to restore the antinociceptive effect of morphine in pertussis toxin (PTX)-treated rats by suppressing spinal microglia activation and inhibiting inflammatory cytokine expression. This study was further investigated the mechanism by which ultra-low dose naloxone promotes analgesia in pertussis toxin-treated rats. MAIN METHODS: Male Wistar rats were implanted with an intrathecal (i.t.) catheter and injected either saline or PTX (1 µg). Four days later, rats randomly received either saline, or ultra-low dose naloxone, or recombinant rat interleukin-10 (rrIL-10) (1 µg) injection followed by saline or morphine (10 µg) 30 min later. In some experiments, mouse anti-rat IL-10 antibody (10 µg) was injected intrathecally into PTX injected rats daily on days 4, 5, 6, and 7. On day 7, ultra-low dose naloxone was given 1h after antibody injection with or without subsequent morphine injection. KEY FINDINGS: PTX injection induced notable thermal hyperalgesia and mechanical allodynia. Injection of ultra-low dose naloxone preserved the antinociceptive effect of morphine in PTX-treated rats and associated an increasing of IL-10 protein expression. Intrathecal injection rrIL-10 alone or in combination with morphine, not only reversed mechanical allodynia but also partially restored the antinociceptive effect of morphine; injection of anti-rat IL-10 antibody attenuated the effect of morphine plus ultra-low dose naloxone on mechanical allodynia and completely inhibited the antinociceptive effect of morphine. SIGNIFICANCE: These results indicate that intrathecal ultra-low dose naloxone induces IL-10 expression in spinal neuron and microglia, which suppresses PTX-induced neuroinflammation and restores the antinociceptive effect of morphine.

Perioperative transient ischemic attack caused by the cessation of warfarin.

This paper describes the circumstances of a patient who had been receiving long-term warfarin treatment, but ceased it prior to surgical operation, sustained a transient ischemic heart attack post-operatively, which eventuated in delayed extubation and locked-in syndrome. For patients at low risk of perioperative bleeding, anticoagulation with oral vitamin K antagonist can probably be able to maintain the therapeutic range (INR ≤ 2.0) extreme. For patients with a high risk of bleeding, the international normalized ratio (INR) should be kept ≤ 1.5. Within this range, patients at low risk of thrombosis can discontinue warfarin treatment for 2-5 days pre-operatively; patients at high risk for thrombosis can stop warfarin but should probably be treated with intravenous or subcutaneous heparin when the INR is subtherapeutic.

Expression of neurotrophic factors in neonatal rats after peripheral inflammation.

UNLABELLED: Neonatal peripheral inflammatory insult might result in the alteration of neuronal development in the nociceptive circuit. During early postnatal period, neurotrophins play important roles in neural development and sensory nerve innervation in the central and peripheral nervous systems. In this study, we investigated mRNA expression for neurotrophic factors and their receptors in the dorsal root ganglia of rat pups during postnatal life after peripheral inflammation induced by injection of complete Freund's adjuvant (CFA) into hind paw on postnatal day 1. Our results showed that mRNA expression levels of alpha-calcitonin gene-related peptides, tropomyosin-related kinase-A (trkA), p75 neurotrophin receptor (p75(NTR)), and brain-derived neurotrophic factor (BDNF) elevated significantly after CFA treatment. Such an increase began 1 day after CFA treatment and lasted 2 to 3 days for trkA, p75(NTR), and BDNF. In contrast, there was no change in mRNA expression levels for neurotrophin-4/5, beta-nerve growth factor (beta-NGF), trkB, glial cell line-derived neurotrophin factor, and receptor protein tyrosine kinase protein. Our study demonstrated that neonatal peripheral inflammatory insult might result in molecular changes of neurotrophic factors, particularly in NGF receptors and BDNF, in the process of neuronal development and plasticity in primary afferents during early neonatal period. PERSPECTIVE: Neonatal peripheral inflammation model has been used for the exploration of neuropathic pain mechanism for years. This work provided further detailed information about possible neurotransmitters and peptides involved in this process. This might also lead to future clinical application.