Simultaneous detection of multiple microRNAs based on fluorescence resonance energy transfer under a single excitation wavelength.

MicroRNAs (miRNAs) play a key role in physiological processes, and their dysregulation is closely related to various human diseases. Simultaneous detection of multiple miRNAs is pivotal to cancer diagnosis at an early stage. However, most multicomponent analyses generally involve multiple excitation wavelengths, which are complicated and often challenging to simultaneously acquire multiple detection signals. In this study, a convenient and sensitive sensor was developed to simultaneously detection of multiple miRNAs under a single excitation wavelength through the fluorescence resonance energy transfer between the carbon dots (CDs)/quantum dots (QDs) and graphene oxide (GO). A hybridization chain reaction (HCR) was triggered by miRNA-141 and miRNA-21, resulting in the high sensitivity with a limit of detection (LOD) of 50 pM (3σ/k) for miRNA-141 and 60 pM (3σ/k) for miRNA-21. This simultaneous assay also showed excellent specificity discrimination against the mismatch. Furthermore, our proposed method successfully detected miRNA-21 and miRNA-141 in human serum samples at a same time, indicating its diagnostic potential in a clinical setting.

Acutalibacter caecimuris sp. nov., Acutalibacter intestini sp. nov. and Neglectibacter caecimuris sp. nov., three novel species of the family Oscillospiraceae isolated from caecal contents of C57BL/6J mice.

The intestines of mice are colonized by diverse, as-yet-uncultivated bacteria. In this report, we describe the isolation, culture, genotypic and phenotypic characterization, as well as taxonomic classification of three novel anaerobic bacterial strains derived from the caecal contents of C57BL/6J male mice. According to the phenotypic and genotype-based polyphasic taxonomy, we propose three novel species within the family Oscillospiraceae. They are Acutalibacter caecimuris sp. nov. (type strain M00118T=CGMCC 1.18042T=KCTC 25739T), Acutalibacter intestini sp. nov. (type strain M00204T=CGMCC 1.18044T=KCTC 25741T) and Neglectibacter caecimuris sp. nov. (type strain M00184T=CGMCC 1.18043T=KCTC 25740T).

Total alkaloid fraction of Leonurus japonicus Houtt. Promotes angiogenesis and wound healing through SRC/MEK/ERK signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Leonurus japonicus Houtt., also known as motherwort, is a traditional Chinese medicine that was first identified in Shennong Bencao Jing, the first and essential pharmacy monograph in China. L. japonicus has been regarded as a good gynecological medicine since ancient times. It has been widely used in clinical settings for treatment of gynecological diseases and postnatal rehabilitation with good efficacy and low adverse effects. AIM OF THE STUDY: The main purpose of this study was to determine the angiogenic and wound healing effects of total alkaloid fraction from L. japonicus Houtt. (TALH) in vivo and in vitro. In addition, the main bioactive components of total alkaloids were to be identified and analyzed in this study. MATERIALS AND METHODS: First, the UHPLC/Q-TOF-MS method was used to identify and quantify the major components in the TALH extract. The wound healing activity was evaluated in vivo using a rat full-thickness cutaneous wound model. Histological study of wound healing in rat model was performed via immunohistochemistry and immunofluorescence. Cell proliferation was determined by MTT assay. Wound healing and transwell assays were used for detection of cell migration. The effect on tube formation was determined by tube formation assay in HUVECs. Western blot and RT-PCR were used to detect the expressions of relative proteins and genes respectively. Knock-down of SRC by siRNA was done to verify the crucial role of SRC in promotion of angiogenesis induced by TALH. RESULTS: Seven characteristic peaks were recognized in the UHPLC/Q-TOF-MS spectrum, while four of the main components were quantified. The wound model in rats showed that treatment of TALH promoted wound healing by stimulating cellular proliferation and collagen deposition. In vitro experiments showed that co-treatment of TALH and VEGF increased cell proliferation, migration and tube formation in HUVECs. Mechanistic studies suggested that the co-treatment increased gene expressions of SRC, MEK1/2 and ERK1/2, as well as the phosphorylation levels of these proteins. Furthermore, the effect of co-treatment was attenuated after SRC knockdown, suggesting that SRC plays an important role in angiogenesis and wound healing induced by TALH and VEGF co-treatment. CONCLUSION: Our results showed that TALH was one of the main active components of L. japonicus that promoted angiogenesis and wound healing by regulating the SRC/MEK/ERK pathway. Our study provided scientific basis for better clinical application of L. japonicas.

High platelet count predicts poor prognosis in HCC patients undergoing TACE: a propensity score-matched analysis.

BACKGROUND: The aim of this study was to retrospectively evaluate the prognostic value of the pretreatment platelet (PLT) count in patients with hepatitis B virus (HBV)-related intermediate-advanced hepatocellular carcinoma (HCC) complicated with cirrhosis undergoing transcatheter arterial chemoembolization (TACE). RESEARCH DESIGN AND METHODS: We assessed 362 patients with HBV-related intermediate-advanced HCC complicated with cirrhosis undergoing TACE. Patients were divided into low (≤96 × 109/L) and high (>96 × 109/L) PLT groups. Propensity score matching (PSM) was performed to eliminate the imbalance in potential confounding factors. The endpoint was time to progression (TTP). RESULTS: After PSM, the high and low PLT groups had 97 patients each. The TTP was significantly longer in the low PLT group than in the high PLT group (log-rank test, p < 0.001). A high pretreatment PLT count was an independent predictor of poor tumor response (OR 4.724; 95% CI 1.889-11.815; P = 0.001) and short TTP (HR = 3.598; 95% CI: 2.570-5.036; P < 0.001). Subgroup analysis showed that a high PLT count increased the risk of progression across almost all subgroups. CONCLUSIONS: The pretreatment PLT count has potential value in predicting the prognosis of patients with intermediate-advanced HCC undergoing TACE.

Differential and paradoxical roles of new-generation antidepressants in primary astrocytic inflammation.

BACKGROUND: Selective serotonin reuptake inhibitors (SSRIs) and serotonin-norepinephrine reuptake inhibitors (SNRIs) are commonly used new-generation drugs for depression. Depressive symptoms are thought to be closely related to neuroinflammation. In this study, we used up-to-date protocols of culture and stimulation and aimed to understand how astrocytes respond to the antidepressants. METHODS: Primary astrocytes were isolated and cultured using neurobasal-based serum-free medium. The cells were treated with a cytokine mixture comprising complement component 1q, tumor necrosis factor α, and interleukin 1α with or without pretreatments of antidepressants. Cell viability, phenotypes, inflammatory responses, and the underlying mechanisms were analyzed. RESULTS: All the SSRIs, including paroxetine, fluoxetine, sertraline, citalopram, and fluvoxamine, show a visible cytotoxicity within the range of applied doses, and a paradoxical effect on astrocytic inflammatory responses as manifested by the promotion of inducible nitric oxide synthase (iNOS) and/or nitric oxide (NO) and the inhibition of interleukin 6 (IL-6) and/or interleukin 1ß (IL-1ß). The SNRI venlafaxine was the least toxic to astrocytes and inhibited the production of IL-6 and IL-1ß but with no impact on iNOS and NO. All the drugs had no regulation on the polarization of astrocytic A1 and A2 types. Mechanisms associated with the antidepressants in astrocytic inflammation route via inhibition of JNK1 activation and STAT3 basal activity. CONCLUSIONS: The study demonstrated that the antidepressants possess differential cytotoxicity to astrocytes and function differently, also paradoxically for the SSRIs, to astrocytic inflammation. Our results provide novel pieces into understanding the differential efficacy and tolerability of the antidepressants in treating patients in the context of astrocytes.

Carbon dots-based fluorescence resonance energy transfer for the prostate specific antigen (PSA) with high sensitivity.

The accurate and sensitive detection of biomarkers has great clinical value for the early diagnosis and treatment of cancer. Due to the excellent optical properties of carbon dots (CDs), CDs-based fluorescent detection methods have attracted increasing attention in bioanalytics. Signal reporters using CDs labeled hairpin DNA, based on Föster resonance energy transfer (FRET), have shown promise for the sensitive detection of biomarkers. In this work, a new method for sensitive biomarker detection using an enzyme-free amplified fluorescence strategy was developed. The strategy was based on FRET between CDs and graphene oxide (GO) combined with catalytic hairpin assembly (CHA). In the absence of the target, the CD-labeled hairpin DNA adsorb onto GO via hydrophobic and π-π stacking interactions, resulting in a FRET quenching of the CDs fluorescence. The introduction of the target could trigger the CHA circuits to form Y-shaped double-stranded DNA (dsDNA), resulting in a recovery of the CD's fluorescence signal. This novel strategy was successfully applied for the selective detection of prostate specific antigen (PSA) with a limit of detection (LOD) of 0.22 ng mL-1 (3σ/k). Additionally, the method could be used to detect carcinoembryonic antigen (CEA) and adenosine triphosphate (ATP) with LOD of 0.56 ng mL-1 (3σ/k) and 80 nM (3σ/k), respectively. Therefore, this work demonstrates a promising way to construct a sensitive and versatile detection platform.

Paroxetine suppresses reactive microglia-mediated but not lipopolysaccharide-induced inflammatory responses in primary astrocytes.

BACKGROUND: Astrocytes are the most abundant glial cells in a brain that mediate inflammatory responses and provide trophic support for neurons. We have previously disclosed that paroxetine, a common selective serotonin reuptake inhibitor, ameliorates LPS-induced microglia activation. However, it remains elusive for the role of paroxetine in astrocytic responses. METHODS: Isolated primary astrocytes were pretreated with paroxetine and stimulated with different stimuli, lipopolysaccharide (LPS) or microglia conditioned medium pre-activated with LPS (M/Lps). Inflammatory and neurotrophic responses, underlying mechanisms and the impact on neuronal survival were assessed. RESULTS: Paroxetine had no impact on LPS-stimulated iNOS, TNF-α, and IL-1ß expression, but inhibited M/Lps-induced TNF-α and IL-1ß expression in primary astrocytes. Paroxetine suppressed M/Lps- but not LPS-induced activation of NF-κB and had no impact on the activation of MAPKs and STAT3. Incubation with the resulted astrocyte conditioned media caused no change in the viability of SH-SY5Y cells. BDNF and MANF mRNA expressions were upregulated by M/Lps and paroxetine, respectively. However, M/Lps- or LPS-induced extracellular releases of NO, TNF-α, and/or BDNF in astrocytes were in minor amount compared to those by microglia. CONCLUSIONS: Paroxetine ameliorates the reactive microglia-mediated inflammatory responses in astrocytes partially via inhibition of the NF-κB pathway but has no impact on LPS-stimulated astrocyte activation. While the effects of paroxetine on secondary astrocytic responses are not robust compared to its effect on the innate immune responses of microglia, the results together may implicate a therapeutic potential of paroxetine against neuroinflammation-associated neurological disorders such as Parkinson's disease.

A mimetic peptide of α2,6-sialyllactose promotes neuritogenesis.

Oxidative stress contributes to the pathogenesis of neurodegenerative diseases. With the aim to find reagents that reduce oxidative stress, a phage display library was screened for peptides mimicking α2,6-sialyllactose (6'-SL), which is known to beneficially influence neural functions. Using Sambucus nigra lectin, which specifically binds to 6'-SL, we screened a phage display library and found a peptide comprising identical sequences of 12 amino acids. Mimetic peptide, reverse peptide and scrambled peptide were tested for inhibition of 6'-SL binding to the lectin. Indeed, lectin binding to 6'-SL was inhibited by the most frequently identified mimetic peptide, but not by the reverse or scrambled peptides, showing that this peptide mimics 6'-SL. Functionally, mimetic peptide, but not the reverse or scrambled peptides, increased viability and expression of neural cell adhesion molecule L1 in SK-N-SH human neuroblastoma cells, and promoted survival and neurite outgrowth of cultured mouse cerebellar granule neurons challenged by H2O2-induced oxidative stress. The combined results indicate that the 6'-SL mimetic peptide promotes neuronal survival and neuritogenesis, thus raising hopes for the treatment of neurodegenerative diseases. This study was approved by the Medical Ethics Committee of Shantou University Medical College, China (approval No. SUMC 2014-004) on February 20, 2014.

Randomized trial on the application of biofragmentable anastomosis ring in intestinal anastomosis.

BACKGROUND: The biofragmentable anastomosis ring (BAR) is a simple alternative device to create intestinal anastomosis. Our study was designed to evaluate the clinical value of BAR in intestinal anastomosis. METHODS: A total of 167 patients performed intestinal anastomosis from January 2002 to February 2006 were randomized to BAR group (n = 82) and manual suture group (n = 85) as control. They were equally allocated to the two groups regarding sex, age, site of anastomosis, emergent or elective surgery and contaminant diseases. The results of postoperative complications and recovery were recorded in each group. RESULTS: Eighty-seven intraperitoneal BAR anastomoses were completed in 82 patients. Two and one postoperative deaths were recorded in BAR and suture group, respectively, no deaths were directly related to anastomotic technique. In suture group, anastomotic leakage and early bleeding both occurred in two patients respectively, no anastomotic bleeding occurred in BAR group, one patient in BAR group developed enterocutaneous fistulae. Perioperative bleeding, operation time and length of hospitalization were similar in two groups (P > 0.05). Time for return of bowel function was significantly shortened in BAR group than that in suture group (P < 0.05). CONCLUSION: The BAR appears to be a standard, easy, safe and effective alternative either in elective or emergent intraperitoneal intestinal anastomotic surgery.