SETD2 regulates chromatin accessibility and transcription to suppress lung tumorigenesis.

SETD2, a H3K36 trimethyltransferase, is the most frequently mutated epigenetic modifier in lung adenocarcinoma, with a mutation frequency of approximately 9%. However, how SETD2 loss of function promotes tumorigenesis remains unclear. Using conditional Setd2-KO mice, we demonstrated that Setd2 deficiency accelerated the initiation of KrasG12D-driven lung tumorigenesis, increased tumor burden, and significantly reduced mouse survival. An integrated chromatin accessibility and transcriptome analysis revealed a potentially novel tumor suppressor model of SETD2 in which SETD2 loss activates intronic enhancers to drive oncogenic transcriptional output, including the KRAS transcriptional signature and PRC2-repressed targets, through regulation of chromatin accessibility and histone chaperone recruitment. Importantly, SETD2 loss sensitized KRAS-mutant lung cancer to inhibition of histone chaperones, the FACT complex, or transcriptional elongation both in vitro and in vivo. Overall, our studies not only provide insight into how SETD2 loss shapes the epigenetic and transcriptional landscape to promote tumorigenesis, but they also identify potential therapeutic strategies for SETD2 mutant cancers.

Low Dimensional Nanostructure-Assisted Long-Range Surface Plasmon Resonance Sensors With High Figure of Merit.

Long-range surface plasmon resonance (LRSPR) sensors have been extensively studied by virtue of their extremely narrow full width at half maxima (FWHM) characteristics, but their low sensitivity remains an important factor limiting the figure of merit (FOM), making the sensors have difficulties in detecting small refractive index changes accurately. To address this problem, this paper proposes and demonstrates a low dimensional nanostructure (Au nanospheres, WS2) assisted LRSPR sensor to achieve an effective enhancement of the sensor interfaced electric field and thus improve the sensitivity. The performance parameters of the two sensors are compared with the LRSPR sensor by finite element method analysis, and the results showed that the assistance of the low dimensional nanostructure has a positive effect on the sensor. The first refractive index sensing experiment of the WS2-assisted LRSPR sensor was realized with a 25.47% increase in sensitivity and a 7.13% increase in FOM simultaneously, and the Au nanospheres-assisted LRSPR sensor with a 29.23% increase in sensitivity and a 15.95% increase in FOM simultaneously. The introduction of low dimensional nanostructures provides a flexible and effective means of sensitization for LRSPR sensors, making the plasmon resonance sensors combine high sensitivity, narrow FWHM and high FOM, which have promising applications in biochemical sensing.

Exosome-shuttled mitochondrial transcription factor A mRNA promotes the osteogenesis of dental pulp stem cells through mitochondrial oxidative phosphorylation activation.

OBJECTIVES: The treatment of bone defects by stem cells (MSCs) has achieved limited success over the recent few decades. The emergence of exosomes provides a new strategy for bone regeneration. Here, we aimed to investigate the effect and mechanisms of exosomes combined with dental pulp stem cells (DPSCs) on bone regeneration. MATERIALS AND METHODS: We isolated exosomes from stem cells from human exfoliated deciduous teeth (SHED) aggregates and evaluated the efficacy of exosomes combined with DPSCs in a cranial bone defect model. The potential mechanisms were further investigated. RESULTS: The effect of exosomes combined with DPSCs was remarkable on bone regeneration in vivo and exosomes promoted osteogenic differentiation of DPSCs in vitro. Mechanistically, exosomes increased the expression of mitochondrial transcription factor A (TFAM) in DPSCs by transferring TFAM mRNA. Moreover, highly expressed TFAM in DPSCs enhanced glutamate metabolism and oxidative phosphorylation (OXPHOS) activity. CONCLUSIONS: Consequently, exosomes strengthened bone regeneration of DPSCs through the activation of mitochondrial aerobic metabolism. Our study provides a new potential strategy to improve DPSC-based bone regenerative treatment.

Plasmonic crescent nanoarray-based surface lattice resonance sensor with a high figure of merit.

Due to the natural accumulation of radiation losses arising from the localization and random arrangement of nanoparticles, the figure of merit (FOM) of localized surface plasmon resonance (LSPR) sensors is usually very low (the value is usually less than 5 RIU-1). However, radiation losses of individual particles will be offset by adjusting the phase of the scattered field which is dependent on the structure parameters of arrays. Based on this, a two-dimensional periodic crescent nanoarray-based surface lattice resonance (SLR) sensor with a high FOM is proposed in this work. Some significant results have been obtained by mode field analysis and adjustment of structural parameters. On the one hand, the line-shape of the SLR spectrum is divided into a Fano-like line and a separate line. And the former usually has an FOM of 101 magnitude while the latter has an FOM of 103 magnitude. On the other hand, the relative size of the excitation wavelengths between SLR and LSPR is also vital. The FOM is higher but resonance depth decreases faster when the relative size increases. In this work, a full width at half-maximum (FWHM) of less than 0.5 nm and FOM of more than 1000 RIU-1 (the quality factor is more than 3000) are achieved by the proposed crescent nanoarrays. In addition, this structure demonstrates that plasmonic nanoarray-based SLR has enormous potential in trace substance detection.

Research advances on surface plasmon resonance biosensors.

The surface plasmon resonance (SPR) phenomenon is of wide interest due to its sensitivity to changes in surface refractive index for the label-free, highly sensitive and rapid detection of biomarkers. This paper reviews research progress on SPR biosensors modified with different substrate structures and surface materials, surface plasmon resonance imaging (SPRI), and SPR-enhanced electrochemiluminescent (ECL) biosensors for applications in biosensing in the last five years. This paper focuses on the research on the application of the SPR phenomenon in the field of bio-detection, reviews the sensing characteristics of SPR biosensors with substrate structures of prisms, gratings, and optical fibers, and summarizes and analyzes the sensitivity and interference resistance of SPR sensors with surface modification of different materials (high-refractive index dielectric films, metallic micro- and nanostructures, and surface antifouling materials). Considering that imaging is an important tool for biomedical detection, this paper reviews the research progress on SPRI technology in the field of biomedical detection. In addition, this paper also reviews the research progress on SPR-enhanced ECL biosensors in the field of biosensing. Finally, this paper provides an outlook on the development trends of biosensing technology in terms of portable high-precision SPR sensors, reduction of self-loss of thin film materials, optimization of image processing techniques and simplification of electrode modification for ECL sensors.

Sensing self-referenced fiber optic long-range surface plasmon resonance sensor based on electronic coupling between surface plasmon polaritons.

A type of hollow gold nanoparticle (HGNP)-modified fiber optic long-range surface plasmon resonance (LRSPR) sensor with sensing self-reference is proposed and demonstrated. HGNPs have a stronger plasmonic field compared to solid GNPs because of the coupling between the inner and outer walls of HGNPs. The intense near-field electronic coupling between long-range surface plasmon polaritons associated with the LRSPR gold layer and localized surface plasmon polaritons of HGNPs leads to localized electromagnetic-field enhancement and LRSPR response signal amplification. Therefore, the HGNP-modified LRSPR sensor possesses a more excellent sensing property compared with the unmodified LRSPR sensor. The long-range resonance dip in the transmission spectrum is shown to shift in response to ambient refractivity change, and the characteristic absorption peak is fixed, allowing to regard it as a reference to improve detection accuracy of the sensors. The mode-field distribution of the sensors is simulated by using the finite element method, and the simulation results show that the electric-field intensity on the HGNP surface is significantly enhanced compared with that of the gold layer surface of the unmodified LRSPR sensor. 1874.79 nm/RIU improvement in sensitivity, 1.42 times improvement in figure of merit (FOM), and approximately 50% reduction in limit of detection (LOD) are achieved for the refractivity measurement of a low-concentration biological solution with the employment of HGNPs in LRSPR sensing experiments. The HGNP-modified LRSPR sensor proposed in this paper has high detection accuracy and FOM and low LOD, and can realize remote real-time online monitoring. Therefore, it has important research value and broad application prospects in the field of biochemical detection.

[Research Progress of Dynamic Musculoskeletal Ultrasound Image Processing].

Muscle is one of the most important tissues of human body, which is distributed around various organs and bones. Skeletal muscle plays an important role in human activities and its functional changes are closely related to its own morphological structure. The study of the relationship between musculoskeletal structure and function can help us to understand the physiology basics of force and to guide clinical practices. Ultrasonography has been widely used in the research of muscle properties since it is real-time, fast, nonradiative and inexpensive. In recent years, there emerges various researches on image processing method for musculoskeletal ultrasonography, especially for dynamic ultrasonography. This paper presents a brief overview of the existing methods and key steps of ultrasound image processing of musculoskeletal.

α-smooth muscle actin is not a marker of fibrogenic cell activity in skeletal muscle fibrosis.

α-Smooth muscle actin (α-SMA) is used as a marker for a subset of activated fibrogenic cells, myofibroblasts, which are regarded as important effector cells of tissue fibrogenesis. We address whether α-SMA-expressing myofibroblasts are detectable in fibrotic muscles of mdx5cv mice, a mouse model for Duchenne muscular dystrophy (DMD), and whether the α-SMA expression correlates with the fibrogenic function of intramuscular fibrogenic cells. α-SMA immunostaining signal was not detected in collagen I (GFP)-expressing cells in fibrotic muscles of ColI-GFP/mdx5cv mice, but it was readily detected in smooth muscle cells lining intramuscular blood vessel walls. α-SMA expression was detected by quantitative RT-PCR and Western blot in fibrogenic cells sorted from diaphragm and quadriceps muscles of the ColI-GFP/mdx5cv mice. Consistent with the more severe fibrosis in the ColI-GFP/mdx5cv diaphragm, the fibrogenic cells in the diaphragm exerted a stronger fibrogenic function than the fibrogenic cells in the quadriceps as gauged by their extracellular matrix gene expression. However, both gene and protein expression of α-SMA was lower in the diaphragm fibrogenic cells than in the quadriceps fibrogenic cells in the ColI-GFP/mdx5cv mice. We conclude that myofibroblasts are present in fibrotic skeletal muscles, but their expression of α-SMA is not detectable by immunostaining. The level of α-SMA expression by intramuscular fibrogenic cells does not correlate positively with the level of collagen gene expression or the severity of skeletal muscle fibrosis in the mdx5cv mice. α-SMA is not a functional marker of fibrogenic cells in skeletal muscle fibrosis associated with muscular dystrophy.

Infiltrating macrophages are broadly activated at the early stage to support acute skeletal muscle injury repair.

Acute skeletal muscle injury repair requires an adequate inflammatory response predominated by macrophage infiltration. We studied the activation of infiltrating macrophages by analyzing the expression of M1/M2 signature genes. Most of the intramuscular macrophages were Ly6Chi at day 1 after BaCl2 injection, while many were Ly6Clo at day 3. Ly6Chi macrophages at day 1 expressed a high level of both M1 and M2 genes, and the Ly6Chi and Ly6Clo macrophages at day 3 expressed a similar level of many M1/M2 genes. Infiltrating macrophages are broadly activated rather than polarized at the early stage to support acute skeletal muscle injury repair.

CCR2 deficiency does not provide sustained improvement of muscular dystrophy in mdx5cv mice.

Genetic ablation or pharmacologic inhibition of CC chemokine receptor type 2 (CCR2) reduced macrophage (MP) infiltration and improved muscle pathology and function in mdx diaphragm muscle at early stages. We addressed whether CCR2 deficiency resulted in sustained improvement of mdx5cv-Ccr2-/- diaphragm. Compared to mdx5cv controls, CCR2 deficiency in mdx5cv-Ccr2-/- mice markedly reduced intramuscular Ly6Chi MPs at all stages, but it reduced Ly6Clow MPs only at early stages (4 and 9 wk). CCR2 deficiency reduced quadriceps and diaphragm muscle damage and fibrosis at 14 wk but not at 6 mo, and it improved diaphragm muscle regeneration and respiratory function at 14 wk but not at 6 mo. Intramuscular MPs in mdx5cv-Ccr2-/- diaphragm expressed a low level of IL-1ß, IL-6, and IFN-γ genes, a similar level of TNF-α, TGF-ß1, and platelet-derived growth factor α genes, and a high level of IGF-1 and osteopontin genes compared to mdx5cv controls. Diaphragm fibroblasts at 14 wk showed a similar cell number with a similar level of collagen and profibrogenic growth factor gene expression in mdx5cv-Ccr2-/- and mdx5cv mice. Diaphragm MPs from both mdx5cv-Ccr2-/- and mdx5cv mice stimulated collagen gene expression by cocultured fibroblasts. The findings suggest that CCR2 deficiency does not provide a sustained benefit and that Ly6Clow MPs may contribute to the progressive fibrosis and dysfunction of mdx5cv diaphragm.-Zhao, W., Wang, X., Ransohoff, R. M., Zhou, L. CCR2 deficiency does not provide sustained improvement of muscular dystrophy in mdx5cv mice.

Identification and Function of Fibrocytes in Skeletal Muscle Injury Repair and Muscular Dystrophy.

We identified and characterized the function of CD45+/collagen I+ fibrocytes in acutely injured skeletal muscle of wild-type (WT) and Ccr2-/- mice, and in quadriceps and diaphragm muscles of mdx5cv mice, a mouse model for Duchenne muscular dystrophy. Fibrocytes were not detected in peripheral blood of WT mice after acute muscle injury or mdx5cv mice. Fibrocytes were detected in acutely injured muscles and in mdx5cv quadriceps and diaphragm muscles. These cells expressed F4/80 and CCR2, and they were mostly Ly6Clo They expressed a low level of collagens but a high level of profibrotic growth factors as compared with i.m. fibroblasts. Fibrocyte expression of collagens and profibrotic growth factors was not increased in Ccr2-/- mice as compared with WT controls. Fibrocyte expression of both proinflammatory and profibrotic cytokines was significantly higher in mdx5cv diaphragm than in mdx5cv quadriceps. In cocultures, fibrocytes from the mdx5cv diaphragm stimulated a higher level of fibroblast expression of extracellular matrix genes than did those from the mdx5cv quadriceps. Our findings suggest that i.m. fibrocytes most likely originate from infiltrating monocytes/macrophages and differentiate within injured muscles. They likely contribute to the normal muscle injury repair by producing growth factors. They do not appear to contribute to the persistent muscle fibrosis associated with poor injury repair in Ccr2-/- mice. However, they likely contribute to the persistent inflammation and progressive fibrosis in the mdx5cv diaphragm.

CX3CR1 deficiency delays acute skeletal muscle injury repair by impairing macrophage functions.

Adequate inflammatory response predominated by macrophage infiltration is essential to acute skeletal muscle injury repair. The majority of intramuscular macrophages express the chemokine receptor CX3CR1. We studied the role of CX3CR1 in regulating intramuscular macrophage number and function in acute injury repair by using a loss-of-function approach. Muscle injury repair was delayed in CX3CR1(GFP/GFP) mice as compared with wild-type (WT) controls. CX3CR1 was predominantly expressed by macrophages but not by myogenic cells or capillary endothelia cells in injured muscles. Intramuscular macrophage number and subset composition were not altered by CX3CR1 deficiency. Intramuscular macrophage phagocytosis function was impaired by CX3CR1 deficiency as demonstrated by increased number of necrotic fibers (+115%) and percentage of necrotic area (+204%) at 7 d, increased number of intramuscular neutrophils at 3 (+89%) but not 1 d, reduced number of phagocytosing macrophages (-12%) and phagocytosed beads within macrophages (-15%) in CX3CR1(GFP/GFP) mice as compared with WT controls. The mRNA expression of CD36 (-50%), CD14 (-43%), IGF-1 (-53%), and IL-6 (-40%) was reduced in CX3CR1-deficient macrophages as compared with WT controls. We conclude that CX3CR1 is important to acute skeletal muscle injury repair by regulating macrophage phagocytosis function and trophic growth factor production.

Beneficial and harmful effects of alcohol exposure on Caenorhabditis elegans worms.

Alcoholic beverages are consumed widely throughout the world. While the harmful effects of alcoholism are well recognized, the beneficial effects of moderate alcohol consumption to human health remain debatable. In this study, we investigated the effects of long-term ethanol exposure on nematode Caenorhabditis elegans worms. At high concentrations (≥ 4%), ethanol significantly impaired mobility, reduced fertility, and shortened lifespan. Interestingly, at low concentrations (1-2%), it extended lifespan, accompanied with a slower decline of mobility during aging, although it slightly impaired development, fertility, and chemotaxis. The lifespan-prolonging effects of ethanol at the low concentrations were seen in normal worms exposed to ethanol from egg, young larva, and young adult stages but were not observed in age-1 and sir-2.1 mutant worms. Our study demonstrated hormetic effects of ethanol and further established C. elegans as a suitable animal model to study ethanol related problems.

Relevance of JAK2V617F positivity to hematological diseases--survey of samples from a clinical genetics laboratory.

BACKGROUND: JAK2V617F is found in the majority of patients with Ph- myeloproliferative neoplasms (MPNs) and has become a valuable marker for diagnosis of MPNs. However, it has also been found in many other hematological diseases, and some studies even detected the presence of JAK2V617F in normal blood samples. This casts doubt on the primary role of JAK2V617F in the pathogenesis of MPNs and its diagnostic value. METHODS: In the present study, we analyzed JAK2V617F positivity with 232 normal blood samples and 2663 patient blood, bone marrow, and amniotic fluid specimens obtained from a clinical genetics laboratory by using a simple DNA extraction method and a sensitive nested allele-specific PCR strategy. RESULTS: We found JAK2V617F present in the majority (78%) of MPN patients and in a small fraction (1.8-8.7%) of patients with other specific hematological diseases but not at all in normal healthy donors or patients with non-hematological diseases. We also revealed associations of JAK2V617F with novel as well as known chromosomal abnormalities. CONCLUSIONS: Our study suggests that JAK2V617F positivity is associated with specific hematological malignancies and is an excellent diagnostic marker for MPNs. The data also indicate that the nested allele-specific PCR method provides clinically relevant information and should be conducted for all cases suspected of having MPNs as well as for other related diseases.

Transgenic expression of JAK2V617F causes myeloproliferative disorders in mice.

The JAK2(V617F) mutation was found in most patients with myeloproliferative disorders (MPDs), including polycythemia vera, essential thrombocythemia, and primary myelofibrosis. We have generated transgenic mice expressing the mutated enzyme in the hematopoietic system driven by a vav gene promoter. The mice are viable and fertile. One line of the transgenic mice, which expressed a lower level of JAK2(V617F), showed moderate elevations of blood cell counts, whereas another line with a higher level of JAK2(V617F) expression displayed marked increases in blood counts and developed phenotypes that closely resembled human essential thrombocythemia and polycythemia vera. The latter line of mice also developed primary myelofibrosis-like symptoms as they aged. The transgenic mice showed erythroid, megakaryocytic, and granulocytic hyperplasia in the bone marrow and spleen, displayed splenomegaly, and had reduced levels of plasma erythropoietin and thrombopoietin. They possessed an increased number of hematopoietic progenitor cells in peripheral blood, spleen, and bone marrow, and these cells formed autonomous colonies in the absence of growth factors and cytokines. The data show that JAK2(V617F) can cause MPDs in mice. Our study thus provides a mouse model to study the pathologic role of JAK2(V617F) and to develop treatment for MPDs.