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SUMMARY Teriparatide is the most widely prescribed bone anabolic drug in the world, but its cellular targets remain incompletely defined. The Gli1+ metaphyseal mesenchymal progenitors (MMPs) are a main source for osteoblasts in postnatal growing mice, but their potential response to teriparatide is unknown. Here, by lineage tracing, we show that teriparatide stimulates both proliferation and osteoblast differentiation of MMPs. Single-cell RNA sequencing reveals heterogeneity among MMPs, including an unexpected chondrocyte-like osteoprogenitor (COP). COP expresses the highest level of Hedgehog (Hh) target genes and the insulin-like growth factor 1 receptor (Igf1r) among all cell clusters. COP also expresses Pth1r and further upregulates Igf1r upon teriparatide treatment. Inhibition of Hh signaling or deletion of Igf1r from MMPs diminishes the proliferative and osteogenic effects of teriparatide. The study therefore identifies COP as a teriparatide target wherein Hh and insulin-like growth factor (Igf) signaling are critical for the osteoanabolic response in growing mice., Graphical Abstract, In brief Gli1+ metaphyseal mesenchymal progenitors (MMPs) are a main source for osteoblasts in postnatal mice. Through single-cell RNA sequencing, Shi et al. find a chondrocyte-like osteoprogenitor (COP) among MMPs. COP is a target of teriparatide, the mainstay of bone anabolic therapy, and requires Hh and Igf signaling for bone formation.

Bone is maintained by coupled activities of bone-forming osteoblasts/osteocytes and bone-resorbing osteoclasts. Alterations in this relationship can lead to pathologic bone loss such as osteoporosis. It is well known that osteogenic cells support osteoclastogenesis via production of RANKL. Interestingly, our recently identified bone marrow mesenchymal cell population-marrow adipogenic lineage precursors (MALPs) that form a multidimensional cell network in bone-was computationally demonstrated to be the most interactive with monocyte-macrophage lineage cells through high and specific expression of several osteoclast regulatory factors, including RANKL. Using an adipocyte-specific Adipoq-Cre to label MALPs, we demonstrated that mice with RANKL deficiency in MALPs have a drastic increase in trabecular bone mass in long bones and vertebrae starting from 1 month of age, while their cortical bone appears normal. This phenotype was accompanied by diminished osteoclast number and attenuated bone formation at the trabecular bone surface. Reduced RANKL signaling in calvarial MALPs abolished osteolytic lesions after LPS injections. Furthermore, in ovariectomized mice, elevated bone resorption was partially attenuated by RANKL deficiency in MALPs. In summary, our studies identified MALPs as a critical player in controlling bone remodeling during normal bone metabolism and pathological bone loss in a RANKL-dependent fashion.

© 2021 The Authors. Osteoarthritis (OA) is a widespread joint disease for which there are no disease-modifying treatments. Previously, we found that mice with cartilage-specific epidermal growth factor receptor (EGFR) deficiency developed accelerated knee OA. To test whether the EGFR pathway can be targeted as a potential OA therapy, we constructed two cartilage-specific EGFR overactivation models in mice by overexpressing heparin binding EGF-like growth factor (HBEGF), an EGFR ligand. Compared to wild type, Col2-Cre HBEGF-overexpressing mice had persistently enlarged articular cartilage from adolescence, due to an expanded pool of chondroprogenitors with elevated proliferation ability, survival rate, and lubricant production. Adult Col2-Cre HBEGF-overexpressing mice and Aggrecan-CreER HBEGF-overexpressing mice were resistant to cartilage degeneration and other signs of OA after surgical destabilization of the medial meniscus (DMM). Treating mice with gefitinib, an EGFR inhibitor, abolished the protective action against OA in HBEGF-overexpressing mice. Polymeric micellar nanoparticles (NPs) conjugated with transforming growth factor-α (TGFα), a potent EGFR ligand, were stable and nontoxic and had long joint retention, high cartilage uptake, and penetration capabilities. Intra-articular delivery of TGFα-NPs effectively attenuated surgery-induced OA cartilage degeneration, subchondral bone plate sclerosis, and joint pain. Genetic or pharmacologic activation of EGFR revealed no obvious side effects in knee joints and major vital organs in mice. Together, our studies demonstrate the feasibility of using nanotechnology to target EGFR signaling for OA treatment.

Meniscal tears are associated with a high risk of osteoarthritis but currently have no disease-modifying therapies. Using Gli1-CreER tdTomato mice, we found that Gli1+ cells contribute to the development of meniscus horns from 2 weeks of age. In adult mice, Gli1+ cells resided at the superficial layer of meniscus and expressed known mesenchymal progenitor markers. In culture, meniscal Gli1+ cells possessed high progenitor activities under the control of Hh signal. Meniscus injury at the anterior horn induced a quick expansion of Gli1+ cells. Normally, the tissue healed slowly, leading to cartilage degeneration. Ablation of Gli1+ cells further hindered this repair process. Strikingly, intra-articular injection of Gli1+ meniscal cells or an Hh activator right after injury accelerated the bridging of the interrupted ends and attenuated signs of osteoarthritis. Taken together, our work identified a novel progenitor population in meniscus and proposes a new treatment for repairing injured meniscus and preventing osteoarthritis.

Bone marrow mesenchymal stem cells (MSCs) are promising therapeutic tools for tissue repair and treatment of a number of human diseases. As a result, there is substantial interest in characterizing and expanding these cells to uncover their therapeutic potential. Bone marrow mesenchymal progenitors, containing both MSCs and their proliferative progeny, are commonly isolated from the central region of rodent long bones. However, challenges exist in expanding these central mesenchymal progenitors in culture. We have designed an enzymatic digestion protocol to isolate mesenchymal progenitors within rodent long bones that resides close to the bone surface, which we termed endosteal mesenchymal progenitors. These cells are more metabolically active and more responsive to external stimuli compared to central mesenchymal progenitors. Therefore, they represent a biologically important target for MSC research. This chapter describes the approach in detail how to isolate and culture endosteal mesenchymal progenitors as well as their central counterparts from rodent long bones.

Bone marrow mesenchymal stem cells (MSCs) are promising therapeutic tools for tissue repair and treatment of a number of human diseases. As a result, there is substantial interest in characterizing and expanding these cells to uncover their therapeutic potential. Bone marrow mesenchymal progenitors, containing both MSCs and their proliferative progeny, are commonly isolated from the central region of rodent long bones. However, challenges exist in expanding these central mesenchymal progenitors in culture. We have designed an enzymatic digestion protocol to isolate mesenchymal progenitors within rodent long bones that resides close to the bone surface, which we termed endosteal mesenchymal progenitors. These cells are more metabolically active and more responsive to external stimuli compared to central mesenchymal progenitors. Therefore, they represent a biologically important target for MSC research. This chapter describes the approach in detail how to isolate and culture endosteal mesenchymal progenitors as well as their central counterparts from rodent long bones.

Bone is maintained by coupled activities of bone-forming osteoblasts/osteocytes and bone-resorbing osteoclasts and an alternation of this relationship can lead to pathologic bone loss such as in osteoporosis. It is well known that osteogenic cells support osteoclastogenesis via synthesizing RANKL. Interestingly, our recently identified bone marrow mesenchymal cell population—marrow adipogenic lineage precursors (MALPs) that form a multi-dimensional cell network in bone—was computationally demonstrated to be the most interactive with monocyte-macrophage lineage cells through highly and specifically expressing several osteoclast regulatory factors, including RANKL. Using an adipocyte-specific Adipoq-Cre to label MALPs, we demonstrated that mice with RANKL deficiency in MALPs have a drastic increase of trabecular bone mass in long bones and vertebrae starting from 1 month of age but that their cortical bone is normal. This phenotype was accompanied by diminished osteoclast number and attenuated bone formation at the trabecular bone surface. Reduced RANKL signaling in calvarial MALPs also abolished osteolytic lesions after lipopolysaccharide (LPS) injections. Furthermore, in ovariectomized mice, elevated bone resorption was partially attenuated by RANKL deficiency in MALPs. In summary, our studies identified MALPs as a critical player in controlling bone remodeling during normal bone metabolism and pathological bone loss in a RANKL-dependent fashion.

Bone marrow mesenchymal lineage cells are a heterogeneous cell population involved in bone homeostasis and diseases such as osteoporosis. While it is long postulated that they originate from mesenchymal stem cells, the true identity of progenitors and their in vivo bifurcated differentiation routes into osteoblasts and adipocytes remain poorly understood. Here, by employing large scale single cell transcriptome analysis, we computationally defined mesenchymal progenitors at different stages and delineated their bi-lineage differentiation paths in young, adult and aging mice. One identified subpopulation is a unique cell type that expresses adipocyte markers but contains no lipid droplets. As non-proliferative precursors for adipocytes, they exist abundantly as pericytes and stromal cells that form a ubiquitous 3D network inside the marrow cavity. Functionally they play critical roles in maintaining marrow vasculature and suppressing bone formation. Therefore, we name them marrow adipogenic lineage precursors (MALPs) and conclude that they are a newly identified component of marrow adipose tissue.

Therapeutic resistance remains a persistent challenge for patients with malignant tumors. Here, we reveal that endothelial cells (ECs) acquire transformation into mesenchymal stem cell (MSC)–like cells in glioblastoma (GBM), driving tumor resistance to cytotoxic treatment. Transcriptome analysis by RNA sequencing (RNA-seq) revealed that ECs undergo mesenchymal transformation and stemness-like activation in GBM microenvironment. Furthermore, we identified a c-Met-mediated axis that induces β-catenin phosphorylation at Ser(675) and Wnt signaling activation, inducing multidrug resistance–associated protein-1(MRP-1) expression and leading to EC stemness-like activation and chemoresistance. Last, genetic ablation of β-catenin in ECs overcome GBM tumor resistance to temozolomide (TMZ) chemotherapy in vivo. Combination of Wnt inhibition and TMZ chemotherapy eliminated tumor-associated ECs, inhibited GBM growth, and increased mouse survival. These findings identified a cell plasticity–based, microenvironment-dependent mechanism that controls tumor chemoresistance, and suggest that targeting Wnt/β-catenin–mediated EC transformation and stemness activation may overcome therapeutic resistance in GBM.

Teriparatide is the most widely prescribed bone anabolic drug in the world, but its cellular targets remain incompletely defined. The Gli1+ metaphyseal mesenchymal progenitors (MMPs) are a main source for osteoblasts in postnatal growing mice, but their potential response to teriparatide is unknown. Here, by lineage tracing, we show that teriparatide stimulates both proliferation and osteoblast differentiation of MMPs. Single-cell RNA sequencing reveals heterogeneity among MMPs, including an unexpected chondrocyte-like osteoprogenitor (COP). COP expresses the highest level of Hedgehog (Hh) target genes and the insulin-like growth factor 1 receptor (Igf1r) among all cell clusters. COP also expresses Pth1r and further upregulates Igf1r upon teriparatide treatment. Inhibition of Hh signaling or deletion of Igf1r from MMPs diminishes the proliferative and osteogenic effects of teriparatide. The study therefore identifies COP as a teriparatide target wherein Hh and insulin-like growth factor (Igf) signaling are critical for the osteoanabolic response in growing mice.

Bone marrow mesenchymal stromal cells are a highly heterogenic cell population containing mesenchymal stem cells as well as other cell types. With the advance of single cell transcriptome analysis, several recent reports identified a prominent subpopulation of mesenchymal stromal cells that specifically express adipocyte markers but do not contain lipid droplets. We name this cell type marrow adipogenic lineage precursor, MALP, and consider it as a major cellular component of marrow adipose tissue. Here, we review the discovery of MALPs and summarize their unique features and regulatory roles in bone. We further discuss how these findings advance our understanding of bone remodeling, mesenchymal niche regulation of hematopoiesis, and marrow vasculature maintenance.

Our purpose is to study the roles of microRNA-338-5p (miR-338-5p) on the proliferation, invasion, and inflammatory response of fibroblast-like synoviocytes (SFs) in rheumatoid arthritis patients by regulating SPRY1. The target relationship between miR-338-5p and SPRY1 was validated through luciferase reporter system. The expression of miR-338-5p and SPRY1 in synovial tissues and synovial cells were detected using RT-PCR and western blot. The mimics and inhibitors of miR-338-5p were transfected into SFs. MTT, Transwell, and ELISA assays were used to analyze cell proliferation, invasiveness, and the secreted extracellular pro-inflammatory cytokines (such as IL-1a, IL-6, COX2) levels of SFs. MiR-338-5p was highly expressed in rheumatoid arthritis tissues and cells, and directly down-regulated the expression of SPRY1 in the SFs of rheumatoid arthritis patients. Cell proliferation, invasiveness and the expression level of pro-inflammatory cytokines in synovial cells increased after the transfection of miR-338-5p mimics, while the proliferation, invasion and expression level of pro-inflammatory cytokines decreased after the transfection of miR-338-5p inhibitors. In conclusion,miR-338-5p promoted the proliferation, invasion and inflammatory reaction in SFs of rheumatoid arthritis by directly down-regulating SPRY1 expression. J. Cell. Biochem. 118: 2295-2301, 2017. © 2017 Wiley Periodicals, Inc.

Bone marrow mesenchymal lineage cells are a heterogeneous cell population involved in bone homeostasis and diseases such as osteoporosis. While it is long postulated that they originate from mesenchymal stem cells (MSCs), the true identity of MSCs and their in vivo bifurcated differentiation routes into osteoblasts and adipocytes remain poorly understood. Here, by employing single cell transcriptome analysis, we identified MSCs and delineated their bi-lineage differentiation paths in young, adult and aging mice. Among several newly discovered mesenchymal subpopulations, one is a distinct population of adipose-lineage cells that we named marrow environment regulating adipose cells (MERAs). MERAs are non-proliferative, post-progenitor cells that express many mature adipocyte markers but are devoid of lipid droplets. They are abundant in the bone marrow of young mice, acting as pericytes and stromal cells that form numerous connections among themselves and with other cells inside bone, including endothelial cells. Genetic ablation of MERAs disrupts marrow vessel structure, promotes de novo bone formation. Taken together, MERAs represent a unique population of adipose lineage cells that exist only in the bone marrow with critical roles in regulating bone and vessel homeostasis.

Objective. IL-35 was reported as a crucial anti-inflammatory cytokine and could efficiently regulate bone metabolism in murine collagen-induced arthritis model. However, the relationship between IL-35 and bone health in human rheumatoid arthritis (RA) has not been clarified. In this study, the aim was to explore the correlations between IL-35 and bone loss in postmenopausal women with RA. Methods. The study included 76 postmenopausal women with RA and 53 healthy postmenopausal women as healthy controls (HCs). Serum IL-35 levels were detected by enzyme-linked immunosorbent assay. Bone mineral density (BMD) at lumbar spine 1-4 and at total hip was measured using dual-energy X-ray absorptiometry. Alkaline phosphatase (ALP), β-isomerised carboxy-terminal cross-linking telopeptide of type I collagen (β-CTX), and 25-(OH) VitD3 were measured by turbidimetric inhibition immunoassay. Results. Serum IL-35 levels were increased compared with HCs, and it positively correlated with BMD and 25-(OH) VitD3 and negatively correlated with β-CTX in postmenopausal women with RA. Furthermore, serum IL-35 levels in the increased ALP group were higher than those in the normal ALP group. Conclusions. IL-35, an important anti-inflammatory cytokine, may participate in the pathogenesis of bone loss in postmenopausal women with RA.

Objective To investigate the effect of interleukin-35 (IL-35) on vascular endothelial growth factor (VEGF) and its receptors, Flt-1 and Flk-1, in a collagen-induced arthritis (CIA) mouse model of rheumatoid arthritis (RA). Methods We established a CIA mouse model and injected IL-35 intraperitoneally. The articular index (AI) was measured based on the amount of erythema, swelling, or joint rigidity and synovial histology was measured by hematoxylin and eosin staining (HE staining). The levels of VEGF, Flt-1, Flk-1, and von Willebrand factor (vWF) expression in CIA synovial tissue were determined by immunohistochemistry. The mRNA and protein expression levels of VEGF, Flt-1, Flk-1, TNF-α, and INF-γ were detected by reverse transcription PCR (RT-PCR) and western blots, respectively. Results The IL-35 treatment decreased the AI and the synovial histological scores of CIA mice. Immunohistochemistry results revealed that the IL-35 treatment downregulated VEGF, Flt-1, Flk-1, and vWF expression in the CIA mice. RT-PCR results showed that the IL-35-treated mice had lower levels of VEGF, Flt-1, Flk-1, and TNF-α mRNA expression than those of the PBS-treated mice. While there was no significant difference in the level of INF-γ mRNA expression between IL-35-treated and PBS-treated mice. Western blot results showed that the IL-35 treatment downregulated the levels of VEGF, Flt-1, Flk-1, and TNF-α in CIA mice, but the level of INF-γ was not significantly affected. Conclusion These findings show that IL-35 may represent a novel therapeutic agent for RA, and the probable mechanisms may rely on inhibiting VEGF and its receptors Flt-1 and Flk-1.

Atrophic nonunion represents an extremely challenging clinical dilemma for both physicians and fracture patients alike, but its underlying mechanisms are still largely unknown. Here, we established a mouse model that recapitulates clinical atrophic nonunion through the administration of focal radiation to the long bone midshaft 2 weeks before a closed, semistabilized, transverse fracture. Strikingly, fractures in previously irradiated bone showed no bony bridging with a 100% nonunion rate. Radiation triggered distinct repair responses, separated by the fracture line: a less robust callus formation at the proximal side (close to the knee) and bony atrophy at the distal side (close to the ankle) characterized by sustained fibrotic cells and type I collagen-rich matrix. These fibrotic cells, similar to human nonunion samples, lacked osteogenic and chondrogenic differentiation and exhibited impaired blood vessel infiltration. Mechanistically, focal radiation reduced the numbers of periosteal mesenchymal progenitors and blood vessels and blunted injury-induced proliferation of mesenchymal progenitors shortly after fracture, with greater damage particularly at the distal side. In culture, radiation drastically suppressed proliferation of periosteal mesenchymal progenitors. Radiation did not affect hypoxia-induced periosteal cell chondrogenesis but greatly reduced osteogenic differentiation. Lineage tracing using multiple reporter mouse models revealed that mesenchymal progenitors within the bone marrow or along the periosteal bone surface did not contribute to nonunion fibrosis. Therefore, we conclude that atrophic nonunion fractures are caused by severe damage to the periosteal mesenchymal progenitors and are accompanied by an extraskeletal, fibro-cellular response. In addition, we present this radiation-induced periosteal damage model as a new, clinically relevant tool to study the biologic basis of therapies for atrophic nonunion. © 2018 American Society for Bone and Mineral Research.

We studied the anti-angiogenic effect of interleukin-35 (IL-35) by investigating its effects on signal transmission through the Janus kinase signal transducer and activator of transcription (JAK-STAT) pathway in fibroblast-like synoviocytes (FLS).Using the collagen-induced arthritis (CIA) model of rheumatoid arthritis (RA), we derived and cultured FLS, stimulated FLS with IL-35 at different concentrations and examined the expression levels of mRNA and protein of both vascular endothelial growth factor (VEGF), fibroblast growth factor-2 (FGF-2), endostatin, TNF-α, and IL-6 using reverse transcription polymerase chain reaction (RT-PCR) and immunoblotting. We used Western blotting to study the effects of IL-35 on the function of the JAK-STAT pathway in FLS.IL-35 treatment inhibited the expression of VEGF, FGF-2, TNF-α and IL-6, and increased the expression of endostatin in FLS. Western blotting showed that IL-35 treatment of CIA-derived FLS resulted in signalling through STAT1, but not through STAT3 or STAT5.IL-35 signalling through STAT1 and inhibition of the expression of mediators of angiogenesis and inflammation in FLS provide a likely mechanism for anti-angiogenic effects seen in experimental models of RA. Our data suggest that IL-35 and its signalling pathway represent a therapeutic target for the treatment of RA and other angiogenesis-related diseases.

Alzheimer's disease (AD) is pathologically characterized by deposition of β-amyloid (Aβ) peptides, which closely correlates with the balance of nerve growth factor (NGF)-related TrkA/p75NTR signaling. (−)-Epigallocatechin-3-gallate (EGCG) is used for prevention and treatment of many neurodegenerative diseases, including AD. However, whether the neuroprotective effects of EGCG treatment were via modulating the balance of TrkA/p75NTR signaling was still unknown. In this study, we found that EGCG treatment (2 mg · kg –1 · day –1) dramatically ameliorated the cognitive impairments, reduced the overexpressions of Aβ(1–40) and amyloid precursor protein (APP), and inhibited the neuronal apoptosis in the APP/PS1 mice. Interestingly, the EGCG treatment enhanced the relative expression level of NGF by increasing the NGF/proNGF ratio in the APP/PS1 mice. Moreover, after EGCG treatment, TrkA signaling was activated by increasing the phosphorylation of TrkA following the increased phosphorylation of c-Raf, ERK1/2, and cAMP response element-binding protein (CREB), simultaneously the p75NTR signaling was significantly inhibited by decreasing the p75ICD expression, JNK2 phosphorylation, and cleaved-caspase 3 expression, so that the Aβ deposits and neuronal apoptosis in the hippocampus were inhibited.

Our purpose is to study the roles of microRNA-338-5p (miR-338-5p) on the proliferation, invasion, and inflammatory response of fibroblast-like synoviocytes (SFs) in rheumatoid arthritis patients by regulating SPRY1. The target relationship between miR-338-5p and SPRY1 was validated through luciferase reporter system. The expression of miR-338-5p and SPRY1 in synovial tissues and synovial cells were detected using RT-PCR and western blot. The mimics and inhibitors of miR-338-5p were transfected into SFs. MTT, Transwell, and ELISA assays were used to analyze cell proliferation, invasiveness, and the secreted extracellular pro-inflammatory cytokines (such as IL-1a, IL-6, COX2) levels of SFs. MiR-338-5p was highly expressed in rheumatoid arthritis tissues and cells, and directly down-regulated the expression of SPRY1 in the SFs of rheumatoid arthritis patients. Cell proliferation, invasiveness and the expression level of pro-inflammatory cytokines in synovial cells increased after the transfection of miR-338-5p mimics, while the proliferation, invasion and expression level of pro-inflammatory cytokines decreased after the transfection of miR-338-5p inhibitors. In conclusion,miR-338-5p promoted the proliferation, invasion and inflammatory reaction in SFs of rheumatoid arthritis by directly down-regulating SPRY1 expression. J. Cell. Biochem. 118: 2295-2301, 2017. © 2017 Wiley Periodicals, Inc.

With the global economic crisis and industrial restructuring, the unemployed are suffering from job loss-related stress and loss of income, which is believed to impair their mental and physical health, while coping and self-efficacy could combat the adverse effects of unemployment on health. Thus, this study aims to describe quality of life (QOL) among unemployed Chinese people and explore the associated factors. A cross-sectional study was conducted by convenience sampling, composed of 1825 unemployed people, from January 2011 to September 2011. Questionnaires pertaining to demographic characteristics, the 36-item Short-Form Health Survey (SF-36), the abbreviated version of the Cope Inventory (Brief COPE) and self-efficacy scales were used to collect information from unemployed people in the eastern, central, and western regions of China. Hierarchical multiple regression analysis was performed to explore the related factors of QOL. A structural equation model (SEM) was used to test the relations among coping, self-efficacy, and QOL. Mental QOL was significantly lower than physical QOL in Chinese unemployed people. Coping had significant effects on both physical component summary (PCS) and mental component summary (MCS), while self-efficacy played the mediating role in the association between Coping and QOL. Unemployed Chinese people’s mental QOL was disrupted more seriously than their physical QOL. An increase in coping could improve QOL by promoting better management of issues brought about by unemployment. In addition, self-efficacy has the ability to reduce the impact of unemployment on QOL, through the mediating path of coping on QOL. This study highlights the need of coping skills training and self-efficacy enhancement for better management of unemployment in order to improve QOL and well-being.

Background Interactions of inflammatory cells with pancreatic cancer cells play crucial roles in pancreatic cancer, however the dynamic changes of inflammatory cell populations in pancreatic cancerogensis and after chemotherapy have not been well eclucidated. The combinational use of aspirin and atrovastatin (Lipitor) have been widely prescribled for cardio-cerebral vascular diseases mainly by regulation of inflammations, and they have been also reported to have plausible anti-tumor effects, however their potential roles in pancreatic cancerogenesis and chemotherapeutic effects have been seldom investigated. We scanned the dynamic changes of pan-inflammatory cell populations in pancreatic cancerogensis and after chemotherapy and found the potential target cell populations. Then we tested the roles of aspirin and Lipitor to regulate these inflammatory cell populations and their effects on pancreatic cancerogenesis and chemotherapeutic effects. Methods Cancerogen, dimethylbenzanthracene (DMBA), was used to induce pancreatic cancerogenesis and subcatunous implantation of syngenic murine Panc02 pancreatic cancer cells was adopted as well. Gemcitabine was used for chemotherapy. The peripheral blood, pancreatic lesions and tumor samples were harvested and analyzed to search for the potential target cell populations. The roles of aspirin and Lipitor to regulate these cell populations and their potential effects on pancreatic cancerogenesis and chemotherapeutic efficacy were investigated both in vitro and in vivo. Results We found progressive accumulations of myeloid-derived suppressor cells (MDSC) and M2-polarzied tumor associated macrophages(M2) in pancreatic lesions accompanied with dynamic reducations of cytotoxic T cells(CTL) and helper T cells(Th) in the progression of pancreatic cancerogenesis. After gemcitabine treatment, the MDSC significantly reduced, however M2 soared up unexpectedly. Aspirin could significantly inhibit the MDSC and M2 to prevent pancreatic cancerogenesis and improve chemotherapeutic effects of gemcitabine, however Lipitor did not significantly affect MDSC, instead it could promote M2 to attenuate the postive effects of aspirin and gemcitabine. Conclusions MDSC and M2 accumulate in progression of pancreatic cancerogenesis and gemcitabine can induce M2. Aspirin could prevent pancreatic cancerogenesis and improve efficacy of gemcitabine partially by inhibiting MDSC and M2, however when used in combination, Lipitor could weaken the efficacy of aspirin and gemcitabine partially by promoting M2. Electronic supplementary material The online version of this article (doi:10.1186/s13046-016-0304-4) contains supplementary material, which is available to authorized users.

The illustration of the procedures of DMBA-induced pancreatic cancerogenesis model. Table S1. The list of antibodies. Table S2. The list of the primers of the real time RT-PCR. (DOCX 2437 kb)

Glucose-regulated protein 78 (GRP78) is a member of the heat-shock protein 70 family. We evaluated the expression of GRP78 using tissue microarray-based immunohistochemistry in tumor tissues and adjacent nontumor tissues from 180 pancreatic ductal adenocarcinoma (PDAC) patients. The associations between the expression levels of GRP78, clinicopathological factors and overall survival were evaluated. The results showed that the expression of GRP78 was significantly higher in PDAC cells than in normal pancreatic duct cells within adjacent nontumor tissues (p in vitro study, the regulation of GRP78 in the PDAC cell lines affected the proliferation, migration and invasion of PDAC cells through the regulation of CyclinD1, cyclin-dependent kinase (CDK) 4, CDK6, phospho-signal transducer, activator of transcription 3 (p-STAT3), janus kinase 2 (JAK2), ras homolog gene family member A (RhoA), Rho-associated kinase 1 (ROCK1) and sterile alpha motif domain containing protein 4 (Smad4). The present data suggest that GRP78 plays a crucial role in the proliferation, migration and invasion of pancreatic cancer cells and may be a suitable prognostic marker in PDAC.

Objective To determine whether interleukin (IL)-33 and soluble ST2 (sST2) are associated with primary Sjogren’s Syndrome (pSS). Methods Serum levels of IL-33 and sST2 in 110 pSS patients and 78 healthy controls were measured by enzyme-linked immunosorbent assay (ELISA). Immunoglobulins, rheumatoid factors (RF), antinuclear antibody (ANA), anti-SSA/RO-52 antibody, anti-SSB antibody and erythrocyte sedimentation rate (ESR) were measured by standard laboratory techniques. Interstitial lung disease (ILD) was identified on high-resolution computed tomography (HRCT). Disease activity in pSS was scored with the European League Against Rheumatism Sjogren’s Syndrome Disease Activity Index (ESSDAI). Results Serum levels of IL-33 and sST2 were significantly elevated in pSS patients, especially in patients with ILD. There was significant positive correlation between IL-33 and RF, anti-SSB antibody. Conclusion IL-33/sST2 may be involved in the pathogenesis of pSS and partly contribute to the ILD in pSS patients.

Hepatoid adenocarcinoma of the stomach (HAS) is a special type of gastric cancer characterized with hepatoid differentiation and the production of large amounts of α-fetoprotein (AFP). The pathogenesis of HAS is still not clear. Most of the relative studies are single case reports, and studies with large sample are absent. The prognosis of HAS is poor. HAS has a high rate of liver metastasis. The biology behaviors of HAS differ from common gastric cancers. Radical resection of the gastric cancer is considered to be the main treatment when no liver metastasis is found, while the treatment regimens of the metastasis lesions are still in debate. The serum AFP test is important for the early detection and diagnosis of HAS, and it is crucial for monitoring the therapeutic effect and the relapse and metastasis of the tumor.

Introduction Interleukin 27 (IL-27), belonging to the IL-12 family, exerts a critical role in immune regulation of infection, autoimmunity, and tumor. In this study, we aimed to investigate the roles of IL-27 and vascular endothelial growth factor (VEGF) in breast cancer. Methods The serum levels of IL-27 and VEGF in 150 patients with breast cancer and 90 control subjects were measured by an enzyme-linked immunosorbent assay. Estrogen receptor, progesterone receptor, and human epithelial growth factor receptor 2 were measured by immunohistochemistry. Clinical stage, tumor size, lymph node metastasis, grade, and histological type were also recorded. Results The serum levels of IL-27 and VEGF were significantly greater in breast cancer patients than in the control group. In addition, there was a significant correlation between IL-27 and VEGF, and serum IL-27 and VEGF levels were associated with the clinical stage of breast cancer. Moreover, the serum levels of IL-27 were especially elevated in breast cancer patients who were estrogen receptor–positive and progesterone receptor–positive. Furthermore, the serum levels of IL-27 and VEGF decreased after patients with breast cancer had modified radical mastectomy. Conclusion Interleukin 27 may be a new prognostic biomarker of breast cancer and a promising target to limit both angiogenesis and tumor growth.

The purpose of this study is to determine whether soluble ST2 (sST2) and interleukin (IL)-33 is involved in dermatomyositis (DM) and polymyositis (PM).Serum sST2 and IL-33 levels in 49 DM and 21 PM were detected by enzyme-linked immunosorbent assay (ELISA). Erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), creatine kinase (CK), lactate dehydrogenase (LDH) and antinuclear antibody (ANA), anti-Jo-1 antibody and anti-Mi-2 antibody were tested by standard laboratory techniques. Interstitial lung disease (ILD) was identified on high-resolution computed tomography (HRCT). The visual analogue scale (VAS) of the disease activity, muscle strength, lung functional parameters and other clinical features of DM/PM patients were recorded as well.Sera sST2 levels were significantly higher in DM and PM patients and correlated with CRP, CK, LDH and VAS. The level of serum sST2 decreased after therapy. Conversely, serum levels of IL-33 in patients with PM and DM were not significantly higher than those from HC.The level of sST2 is elevated in sera of DM and PM patients. sST2 levels were correlated with other markers of disease activity. This data support that sST2 may play a role in DM and PM.

Background Alzheimer’s disease (AD), an age-related neurodegenerative disorder, is the predominant form of dementia in the elderly, clinically characterized by cognitive impairment and pathologically characterized by extracellular senile plaques largely composed of b-amyloid (Ab) peptide. The overburden of b-amyloid (Ab )d eposition in AD is associated with peroxidation which plays an important role in neuronal dysfunction. APP/PS1 double transgenic mice, which display significant learning and memory impairments and the typical pathological changes such as numerous Ab deposition, are ideal model to mimic the symptoms of AD. (-)-Epigallocatechin-3-gallate (EGCG) is the most abundant polyphenolic constituent in green tea, which has ironchelating, anti-inflammatory, antioxidant and anticancer activities. However, it remains unclear whether EGCG improves learning and memory impairments by attenuating peroxidation in APP/PS1 double transgenic mice. Therefore, we evaluated the relationships between the ameliorated memory dysfunctions and the inhibited peroxidative levels in APP/PS1 transgenic mice after EGCG treatment. Methods APP/PS1 mice at the age of 9 months, randomly distributed into EGCG-treated APP/PS1 group (EGCG group) or APP/PS1 group, and age-matched wild-type mice (C57 BL/6J, WT group) were assigned as aging control. Mice were intragastrically administered EGCG (2 mg/ kg) or vehicle (distilled water) once daily for 4 weeks. Memory function was evaluated in Morris Water Maze (MWM) and passive avoidance test (PAT). The contents of malondialdehyde (MDA), and the activities of total superoxide dismutase (t-SOD) and glutathione peroxidase (GSH-Px) in hippocampus were examined using lipid preoxidation MDA assay kit, total superoxide dismutase assay kit with WST-1, and total glutathione peroxidase assay kit, respectively. The protein expression of inducible nitric-oxide synthase (iNOS) in the hippocampus was measured by Western blot. Results In PAT, the shorter latency and the increased error time to enter the dark compartment of APP/PS1 group were found compared with WT group (P

Background Besides the rapid growth of economy, unemployment becomes a severe socio-economic problem in China. The huge population base in China makes the unemployed population a tremendously huge number. However, health status of unemployed population was ignored and few studies were conducted to describe the depressive symptoms of unemployed individuals in China. This study aims to examine the relationship between Big five personality and depressive symptoms and the mediating role of self-efficacy in this relationship. Methods This cross-sectional study was performed during the period of July to September 2011. Questionnaires consisting of the Center for Epidemiologic Studies Depression Scale (CES-D), the Big Five Inventory (BFI) and the General Self-efficacy Scale (GSE), as well as demographic factors, were used to collect information of unemployed population. A total of 1,832 individuals (effective response rate: 73.28%) became our subjects. Hierarchical linear regression analyses were performed to explore the mediating role of self-efficacy. Results The prevalence of depressive symptoms was 67.7% among Chinese unemployed individuals. After adjusting for demographic characteristics, extraversion, agreeableness and conscientiousness were all negatively associated with depressive symptoms whereas neuroticism was positively associated with depressive symptoms. The proportion of mediating effect of self-efficacy in the relationship between extraversion/agreeableness/conscientiousness/neuroticism and depressive symptoms was 25.42%, 10.91%, 32.21% and 36.44%, respectively. Self-efficacy is a mediator in the relationship between extraversion/agreeableness/conscientiousness/neuroticism and depressive symptoms. Conclusion Self-efficacy partially mediated the relationship between Big five personality and depressive symptoms among Chinese unemployed individuals. Interventions that focus on both individuals’ personality and self-efficacy may be most successful to reduce depressive symptoms of unemployed individuals.

Background ST2, a member of the interleukin (IL)-1receptor family, regulates Th1/Th2 immune responses in autoimmune and inflammatory conditions. However, the role of ST2 signaling in tumor growth and metastasis of breast cancers has not been investigated. This study investigated the possible role of soluble ST2 (sST2) in breast cancer. Methods The serum levels of IL-33, sST2, and vascular endothelial growth factor (VEGF) in 150 breast cancer patients and 90 healthy women were measured by enzyme-linked immunosorbent assay. Estrogen receptor(ER), progesterone receptor, human epithelial receptor (HER)-2, and cell cycle regulated protein Ki-67 were measured. Clinical stage, tumor size, lymph node metastasis, and histological type were also recorded. Results The serum levels of sST2, IL-33, and VEGF were significantly higher in breast cancer patients than in the control group (P P r = 0.375, P r = 0.164, P = 0.045). Serum levels of sST2, IL-33, and VEGF decreased after modified radical mastectomy in ER-positive breast cancers. Serum levels of IL-33, sST2, and VEGF and clinicopathological factors were not significantly correlated with disease-free survival and overall survival of ER-positive breast cancer women during follow-up. Conclusion Serum sST2 levels in ER-positive breast cancer patients are significantly associated with factors that indicate poor prognosis.