Amphiregulin promotes cardiac fibrosis post myocardial infarction by inducing the endothelial-mesenchymal transition via the EGFR pathway in endothelial cells.

The endothelial-mesenchymal transition (EndMT) plays a key role in the development of cardiac fibrosis (CF) after acute myocardial infarction (AMI). The results of our previous study showed that amphiregulin (AR) expression was enhanced after MI. However, the role of AR on EndMT post MI remains unknown. This study aimed to elucidate the impact of AR on EndMT post MI and the associated molecular mechanisms. AR expression was markedly enhanced in infarct border area post MI, and endothelial cells were one of the primary cell sources of AR secretion. Stimulation with AR promoted endothelial cell proliferation, invasion, migration, collagen synthesis and EndMT. In addition, EGFR and downstream gene expression was significantly enhanced. In vivo, EndMT was significantly inhibited after lentivirus-AR-shRNA was delivered to the myocardium post MI. In addition, silencing AR ameliorated cardiac function by decreasing the extent of CF. Furthermore, the levels of EGFR pathway components in endothelial cells extracted from infarct border myocardium were all significantly decreased in lentivirus-AR-shRNA-treated MI mice. Our results demonstrate that AR induces CF post MI by enhancing EndMT in endothelial cells. Thus, targeting the regulation of AR may provide a potentially novel therapeutic option for CF after MI.

Amphiregulin enhances cardiac fibrosis and aggravates cardiac dysfunction in mice with experimental myocardial infarction partly through activating EGFR-dependent pathway.

Cardiac fibrosis (CF), a main process of ventricular remodeling after myocardial infarction (MI), plays a crucial role in the pathogenesis of heart failure (HF) post-MI. It is known that amphiregulin (AR) is involved in fibrosis of several organs. However, the expression of AR and its role post-MI are yet to be determined. This study aimed to investigate the impact of AR on CF post-MI and related mechanisms. Significantly upregulated AR expression was evidenced in the infarct border zone of MI mice in vivo and the AR secretion was enhanced in macrophages, but not in cardiac fibroblasts. In vitro, treatment with AR increased cardiac fibroblast migration, proliferation and collagen synthesis, and upregulated the expression of epidermal growth factor receptor (EGFR) and the downstream genes such as Akt, ERK1/2 and Samd2/3 on cardiac fibroblasts. All these effects could be abrogated by pretreatment with a specific EGFR inhibitor. To verify the functions of AR in MI hearts, lentivirus-AR-shRNA and negative control vectors were delivered into the infarct border zone. After 28 days, knock-down of AR increased the survival rate and improved cardiac function, while decreasing the extent of myocardial fibrosis of MI mice. Moreover, EGFR and the downstream genes were significantly downregulated in lentivirus-AR-shRNA treated MI mice. Our results thus indicate that AR plays an important role in promoting CF after MI partly though activating the EGFR pathway. Targeting AR might be a novel therapeutic option for attenuating CF and improve cardiac function after MI.

Successful targeting of the NRG1 pathway indicates novel treatment strategy for metastatic cancer.

BACKGROUND: NRG1 fusion-positive lung cancers have emerged as potentially actionable events in lung cancer, but clinical support is currently limited and no evidence of efficacy of this approach in cancers beyond lung has been shown. PATIENTS AND METHODS: Here, we describe two patients with advanced cancers refractory to standard therapies. Patient 1 had lung adenocarcinoma and patient 2 cholangiocarcinoma. Whole-genome and transcriptome sequencing were carried out for these cases with select findings validated by fluorescence in situ hybridization. RESULTS: Both tumors were found to be positive for NRG1 gene fusions. In patient 1, an SDC4-NRG1 gene fusion was detected, similar gene fusions having been described in lung cancers previously. In patient 2, a novel ATP1B1-NRG1 gene fusion was detected. Cholangiocarcinoma is not a disease type in which NRG1 fusions had been described previously. Integrative genome analysis was used to assess the potential functional significance of the detected genomic events including the gene fusions, prioritizing therapeutic strategies targeting the HER-family of growth factor receptors. Both patients were treated with the pan HER-family kinase inhibitor afatinib and both displayed significant and durable response to treatment. Upon progression sites of disease were sequenced. The lack of obvious genomic events to describe the disease progression indicated that broad transcriptomic or epigenetic mechanisms could be attributed to the lack of prolonged response to afatinib. CONCLUSION: These observations lend further support to the use of pan HER-tyrosine kinase inhibitors for the treatment of NRG1 fusion-positive in both cancers of lung and hepatocellular origin and indicate more broadly that cancers found to be NRG1 fusion-positive may benefit from such a clinical approach regardless of their site of origin. CLINICAL TRIAL INFORMATION: Personalized Oncogenomics (POG) Program of British Columbia: Utilization of Genomic Analysis to Better Understand Tumour Heterogeneity and Evolution (NCT02155621).

Exosomes Derived from Mesenchymal Stem Cells Rescue Myocardial Ischaemia/Reperfusion Injury by Inducing Cardiomyocyte Autophagy Via AMPK and Akt Pathways.

BACKGROUND/AIMS: Reperfusion after an ischaemic insult might cause infarct extension. Mesenchymal stem cell (MSC)-derived exosomes could attenuate myocardial remodelling in animal models of myocardial ischaemia reperfusion injury (MIRI), and the present study aimed to explore the related mechanisms. METHODS: In vitro, rat H9C2 cardiomyocytes (H9C2s) were exposed to H2O2. Cell viability was detected by the CCK-8 assay, apoptosis was detected by Annexin V-PE/7-AAD staining, ROS production was detected by fluorescence microscopy and flow cytometry, and apoptosis-related proteins and signalling pathway-related proteins were detected by western blot analysis. Autophagic flux was measured using the tandem fluorescent mRFG-GFP-LC3 assay. MSC-derived exosomes were extracted using the total exosome isolation reagent. Apoptosis, myocardial infarction size, heart function and myocardial LC3B expression were examined in an in vivo I/R model by the TUNEL assay, TTC/Evan blue staining, echocardiography and immunohistochemicalstaining, respectively. RESULTS: In vitro, H2O2 dose-dependently increased ROS production and cell apoptosis in H9C2s and blocked autophagic flux after 3 h of exposure; autophagy gradually decreased thereafter, and the lowest level was detected at 12 h after exposure. MSC-derived exosomes reduced H2O2-induced ROS production and cell apoptosis and enhanced autophagy at 12 h after exposure. In H9C2 cells exposed to H2O2 for 12 h, treatment with exosomes enhanced autophagy via the AMPK/mTOR and Akt/mTOR pathways. Likewise, in vivo exosome injections in rats that underwent I/R injury significantly reduced apoptosis and the myocardial infarct size and upregulated myocardial LC3B expression as well as improved heart function. CONCLUSIONS: Our results indicate that MSC-derived exosomes could reduce MIRI by inducing cardiomyocyte autophagy via AMPK/mTOR and Akt/mTOR pathways.

Enhancement of Anti-Hypoxic Activity and Differentiation of Cardiac Stem Cells by Supernatant Fluids from Cultured Macrophages that Phagocytized Dead Mesenchymal Stem Cells.

BACKGROUND: Most mesenchymal stem cells (MSCs) die shortly after transplantation into a myocardial infarcted area. Dead MSCs (dMSCs) are phagocytized by macrophages (pMΦ) in vivo and in vitro; however, the effects of pMΦ on cardiac stem cells (CSCs) remain unknown. METHODS: MSCs, CSCs, and macrophages were obtained from bone marrow, hearts, and peritoneal cavity of mice, respectively. dMSCs were harvested after hypoxia for 24 h, and incubated with macrophages (2:1) for another 2 days with or without lipopolysaccharide (LPS, 50 ng/mL) and sorted by flow cytometry to obtain pMΦ. Viability and apoptosis of CSCs were respectively evaluated with the cell counting kit-8 (CCk-8) assay and Annexin V-PE/7-AAD staining at 0, 6, 12, and 24 h of culture with supernatant fluids from macrophages (MΦ), LPS-stimulated macrophages (LPS-pMΦ), pMΦ, and MSCs. GATA-4 and c-TnI expression was measured by flow cytometry on the seventh day. Expression of inflammation and growth factors was assessed by real-time polymerase chain reaction (RT-PCR) in MΦ, LPS-pMΦ, and pMΦ cells. RESULTS: pMΦ expressed higher levels of interleukin-10 (IL-10) and transforming growth factor-ß (TGF-ß)and lower levels of tumor necrosis factor-α(TNF-α)and IL-6 than LPS-pMΦ, higher levels of growth factors and of GATA-4 and c-TnI at the 7th day, which were similar to those in MSCs. CSCs cultured with supernatant fluids of pMΦ exhibited higher proliferative, anti-hypoxic, and differentiation activities. CONCLUSION: The supernatant fluids of macrophages that had phagocytized dead MSCs encouraged changes in phenotype and growth factor expression, enhanced proliferation, differentiation, and anti-hypoxic activity of CSCs, which is relevant to understanding the persistent therapeutic effect of MSCs after their massive demise upon transplantation in myocardial infarction. Furthermore, some miRNAs or proteins which were extracted from the supernatant fluids may give us a new insight into the treatment of myocardial infarction in the future.

Cryptic diversity begets challenges and opportunities in biodiversity research.

How many species of life are there on Earth? This is a question that we want to know but cannot yet answer. Some scholars speculate that the number of species may reach 2.2 billion when considering cryptic diversity and that each morphology-based insect species may contain an average of 3.1 cryptic species. With nearly two million described species, such high estimates of cryptic diversity would suggest that cryptic species are widespread. The development of molecular species delimitation has led to the discovery of a large number of cryptic species, and cryptic biodiversity has gradually entered our field of vision and attracted more attention. This paper introduces the concept of cryptic species, how they evolve, and methods by which they may be discovered and confirmed, and provides theoretical and methodological guidance for the study of hidden species. A workflow of how to confirm cryptic species is provided. In addition, the importance and reliability of multi-evidence-based integrated taxonomy are reaffirmed as a way to better standardize decision-making processes. Special focus on cryptic diversity and increased funding for taxonomy is needed to ensure that cryptic species in hyperdiverse groups are discoverable and described. An increased focus on cryptic species in the future will naturally arise as more difficult groups are studied, and thereby, we may finally better understand the rules governing the evolution and maintenance of cryptic biodiversity.

Cenozoic Tethyan changes dominated Eurasian animal evolution and diversity patterns.

Cenozoic tectonic evolution in the Tethyan region has greatly changed the landforms and environment of Eurasia, driving the evolution of animals and greatly affecting the diversity patterns of Eurasian animals. By combining the latest Tethyan paleogeographic models and some recently published Eurasian zoological studies, we systematically summarize how tectonic evolution in the Tethyan region has influenced the evolution and diversity patterns of Eurasian animals. The convergence of continental plates, closure of Tethys Sea, and Tethyan sea-level changes have directly affected the composition and spatial distribution of Eurasian animal diversity. The topographic and environmental changes caused by Tethyan tectonics have determined regional animal diversity in Eurasia by influencing animal origin, dispersal, preservation, diversification, and extinction. The ecological transformations resulted in the emergence of new habitats and niches, which promoted animal adaptive evolution, specialization, speciation, and expansion. We highlight that the Cenozoic tectonic evolution of the Tethyan region has been responsible for much of the alteration in Eurasian animal distribution and has been an essential force in shaping organic evolution. Furthermore, we generalize a general pattern that Tethyan geological events are linked with Eurasian animal evolution and diversity dynamics.

Biological and extrinsic correlates of extinction risk in Chinese lizards.

China is a country with one of the most species-rich reptile faunas in the world. However, nearly a quarter of Chinese lizard species assessed by the China Biodiversity Red List are threatened. Nevertheless, to date, no study has explicitly examined the pattern and processes of extinction and threat in Chinese lizards. In this study, we conducted the first comparative phylogenetic analysis of extinction risk in Chinese lizards. We addressed the following 3 questions: (1) What is the pattern of extinction and threat in Chinese lizards? (2) Which species traits and extrinsic factors are related to their extinction risk? (3) How can we protect Chinese lizards based on our results? We collected data on 10 species traits (body size [BS], clutch size, geographic range size, activity time, reproductive mode, habitat specialization [HS], habitat use, leg development, maximum elevation, and elevation range) and 7 extrinsic factors (mean annual precipitation (MAP), mean annual temperature, mean annual solar insolation, normalized difference vegetation index (NDVI), human footprint, human population density, and human exploitation). After phylogenetic correction, these variables were used separately and in combination to assess their associations with extinction risk. We found that Chinese lizards with a small geographic range, large BS, high HS, and living in high MAP areas were vulnerable to extinction. Conservation priority should thus be given to species with the above extinction-prone traits so as to effectively protect Chinese lizards. Preventing future habitat destruction should also be a primary focus of management efforts because species with small range size and high HS are particularly vulnerable to habitat loss.

Clinical observation and analysis of thyroid hormone levels in patients with idiopathic membranous nephropathy.

We sought to investigate the effect of total triiodothyronine (TT3) reduction in the follow-up of patients with idiopathic membranous nephropathy (IMN). A total of 121 patients were enrolled and classified into a low TT3 group or a normal group. Clinical indicators were compared between the groups, and changes in estimated glomerular filtration rate (eGFR), albumin (ALB), thyroid-stimulating hormone, serum creatinine, total protein, total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) during follow-up were analysed. In the analysis by TT3 level, ALB was significantly lower in the low TT3 group (P < .05), while TC, TG, LDL-C, fibrinogen, and renal pathological staging were significantly higher in the low TT3 group (P < .05). Analysis of variance for repeated measurement during follow-up showed that there were no significant differences in eGFR and ALB between the groups. TC, TG, and LDL-C levels were significantly higher in the low TT3 group (P < .05). Approximately 37% of patients with IMN showed a decrease in TT3, which was accompanied by significantly decreased ALB level, higher pathological stage, and increased serum lipid level compared with patients having a normal TT3 level. The management of TT3, and appropriate intervention, may therefore help to prevent the kidney damage progress in patients with IMN.

Why are rates of sterilization in decline? A pilot study designed to explore reasons for declining female sterilization in Scotland.

BACKGROUND: In the last decade, female sterilization had been in decline throughout the UK. It is not clear whether fewer women are requesting sterilization or whether the universal enthusiasm for long-acting reversible methods is leading health professionals to discourage women from being sterilized. Since correct and consistent use of alternative, reversible contraceptive methods depends somewhat on their acceptability, it is important to determine whether women are being refused sterilization or whether they are freely choosing other methods. This study aims to explore whether female sterilization is being widely considered as a contraceptive method, the reasons for choosing or rejecting it, and whether women are being discouraged by health professionals from being sterilized. STUDY DESIGN: A self-completed questionnaire survey among 205 women aged 30 to 50 years who felt that their family was complete attending a family planning clinic in Scotland. RESULTS: Of the 203 women included in the study, 151 (74.4%) had heard of female sterilization, 90 had discussed it with someone (60%) and 87 (58%) had considered it as a contraceptive option. Of the 56 women who consulted their family doctor about sterilization, almost half (27; 48%) were not referred to a hospital and fewer than one (17, 30.4%) in three of them was eventually sterilized or had arrangements in place to get it done. Free-text comments from the women revealed a variety of reasons for not choosing female sterilization and suggested that some women are being deterred from sterilization. CONCLUSION: The study suggests that some women are being actively encouraged by health professionals to use long-acting reversible contraceptive methods and discouraged from choosing sterilization. However, other women recognize for themselves the wisdom of keeping their fertility options open.

Phylogenetic relationships within the genus Jesogammarus (Crustacea, Amphipoda, Anisogammaridae) deduced from mitochondrial COI and 12S sequences.

The genus Jesogammarus contains 16 species in two subgenera, Jesogammarus and Annanogammarus. To examine relationships among species in the genus, a molecular phylogenetic study including eight species of the former subgenus and four of the latter was conducted using partial DNA sequences of the mitochondrial COI and 12S rRNA genes. MP, NJ, and ML trees based on the combined COI and 12S data indicated monophyly of the subgenus Annanogammarus, though the monophyly of Jesogammarus was left unresolved. Consistent with few morphological differences, Jesogammarus (A.) naritai and J. (A.) suwaensis showed low genetic differentiation and did not show reciprocal monophyly, which suggests a close affinity of these taxa.

Myanmarorchestia victoria sp. nov. (Crustacea, Amphipoda, Talitridae), a new species of landhopper from the high altitude forests in Myanmar.

Myanmarorchestia victoriasp. nov. is described from high altitude habitats in Myanmar. The new species differs morphologically from its congeners by palp of maxilliped narrow; sexually dimorphic gnathopod Ⅱ, propodus of male chelate and propodus of female mitten-shaped; and dimorphic uropod Ⅱ, outer ramus of male with small teeth distally, outer ramus of female with three distal spines. Analysis of DNA barcode sequences and niche distinctiveness support recognition of the new species.

Quick starting hormonal contraception after using oral emergency contraception: a systematic review.

INTRODUCTION: Unprotected intercourse after oral emergency contraception (EC) significantly increases pregnancy risk. This underlies the importance of promptly starting effective, ongoing contraception - known as 'quick starting'. However, theoretical concern exists that quick starting might interact with EC or hormonal contraception (HC) potentially causing adverse side effects. METHOD: A systematic review was conducted, evaluating quick starting HC after oral EC [levonorgestrel 1.5 mg (LNG) or ulipristal acetate 30 mg (UPA)]. PubMed, EMBASE, The Cochrane Library, ICTRP, ClinicalTrials.gov and relevant reference lists were searched in February 2016. A lack of comparable studies prevented meta-analysis. RESULTS: Three randomised controlled trials were identified. Two biomedical studies suggested HC action was unaffected by quick starting after UPA; one study examined ovarian quiescence (OR 1.27; 95% CI 0.51-3.18) while taking combined oral contraception (COC). Another assessed cervical mucus impenetrability (OR 0.76; 95% CI 0.27-2.13) while taking progestogen-only pills (POP). Quick starting POP reduced the ability of UPA to delay ovulation (OR 0.04; 95% CI 0.01-0.37). Side effects (OR 1.22; 95% CI 0.48-3.12) and unscheduled bleeding (OR 0.53; 95% CI 0.16-1.81) were unaffected by quick starting COC after UPA. Another study reported higher self-reported contraceptive use at 8 weeks among women quick starting POP after LNG, compared with women given LNG alone (OR 6.73; 95% CI 2.14-21.20).

Views of general practitioners on providing contraceptive advice and long-acting reversible contraception at the 6-week postnatal visit: a qualitative study.

BACKGROUND: Increased uptake of long-acting reversible contraception (LARC) postpartum could prevent more unintended pregnancies and short inter-birth intervals. General practitioners (GPs) play a pivotal role in providing postpartum contraception at the 6-week postnatal visit. AIM: To explore how GPs view their role in delivering postpartum contraception at the 6-week visit and on providing LARC at this time. METHODS: In-depth, semi-structured interviews with a purposive sample of 13 GPs in Edinburgh and the surrounding region in Scotland. The interviews were audio recorded, transcribed and thematically analysed. RESULTS: All GPs confirmed that contraception was routinely discussed at the postpartum visit, although this was usually the last item covered. Most felt that while 6-weeks postpartum was adequate for most women to commence contraception, it was often too late for young mothers (aged under 20 years) or women from deprived areas. GPs provided prescriptions for oral contraception at this visit, but insertion of a contraceptive implant required a further appointment. For intrauterine contraception, women typically required two additional visits to the GP (for counselling and then insertion) or were referred to a local sexual health service. Some GPs saw their role as the main provider of postpartum contraception, whereas others felt they complemented the actions of midwives and health visitors. CONCLUSIONS: This study demonstrated that although contraception is discussed at a routine 6-week postpartum visit with a GP, there are delays for women wishing to commence LARC that create scope for unintended pregnancy. Strategies to facilitate access to LARC postpartum should be explored.

Metformin Inhibits Advanced Glycation End Products-Induced Inflammatory Response in Murine Macrophages Partly through AMPK Activation and RAGE/NFκB Pathway Suppression.

Advanced glycation end products (AGEs) are major inflammatory mediators in diabetes, affecting atherosclerosis progression via macrophages. Metformin slows diabetic atherosclerosis progression through mechanisms that remain to be fully elucidated. The present study of murine bone marrow derived macrophages showed that (1) AGEs enhanced proinflammatory cytokines (interleukin-1ß (IL-1ß), IL-6, and tumor necrosis factor-α (TNF-α)) mRNA expression, RAGE expression, and NFκB activation; (2) metformin pretreatment inhibited AGEs effects and AGEs-induced cluster designation 86 (CD86) (M1 marker) expression, while promoting CD206 (M2 marker) surface expression and anti-inflammatory cytokine (IL-10) mRNA expression; and (3) the AMPK inhibitor, Compound C, attenuated metformin effects. In conclusion, metformin inhibits AGEs-induced inflammatory response in murine macrophages partly through AMPK activation and RAGE/NFκB pathway suppression.

Pioglitazone alleviates inflammation in diabetic mice fed a high-fat diet via inhibiting advanced glycation end-product-induced classical macrophage activation.

Classically activated macrophages (M1) are associated with inflammation in diabetic patients. Inflammation is a known risk factor in diabetes. The present study tested the hypothesis that pioglitazone (PIO) alleviates inflammation in diabetic mice fed a high-fat diet by inhibiting advanced glycation end-product (AGE)-induced classical macrophage activation. It was found that AGE treatment promoted the transcription of pro-inflammatory molecules and M1 surface markers, whereas PIO increased the expression of anti-inflammatory genes and decreased the expression of pro-inflammatory mediators in bone marrow-derived macrophages (BMDMs) in a dose-dependent manner. Furthermore, pretreatment with PIO abrogated the effects of AGE on pro-inflammatory markers and partly inhibited AGE-induced nuclear factor-κB (NF-κB) activation. PIO treatment partly reduced the inflammatory phenotype in diabetic ApoE(-/-) mice, and significantly reduced NF-κB activation in plaques. Therefore, we conclude that PIO blocks classical activation of macrophages and attenuates inflammation in mouse models of diabetes.

Advanced Glycation End Products Enhance Macrophages Polarization into M1 Phenotype through Activating RAGE/NF-κB Pathway.

Atherosclerotic lesions are accelerated in patients with diabetes. M1 (classically activated in contrast to M2 alternatively activated) macrophages play key roles in the progression of atherosclerosis. Since advanced glycation end products (AGEs) are major pathogenic factors and active inflammation inducers in diabetes mellitus, this study assessed the effects of AGEs on macrophage polarization. The present study showed that AGEs significantly promoted macrophages to express IL-6 and TNF-α. M1 macrophage markers such as iNOS and surface markers including CD11c and CD86 were significantly upregulated while M2 macrophage markers such as Arg1 and CD206 remained unchanged after AGEs stimulation. AGEs significantly increased RAGE expression in macrophages and activated NF-κB pathway, and the aforementioned effects were partly abolished by administration of anti-RAGE antibody or NF-κB inhibitor PDTC. In conclusion, our results suggest that AGEs enhance macrophage differentiation into proinflammatory M1 phenotype at least partly via RAGE/NF-κB pathway activation.

Three new species of the genus Gammarus (Crustacea, Amphipoda, Gammaridae) from Yunnan, China.

Three new species of the genus Gammarus are described from Lijiang, Yunnan Province, South China. Gammarus elevatus sp. nov. is characterized by mid-dorsal keel on pleonites 1-3 and compressed elevation on urosomites 1-2; G. denticulatus sp. nov. by many small spinules and setae on pleonites 1-3; G. stagnarius sp. nov. by lack of calceoli on antenna 2 and shorter inner ramus of uropod 3. These amphipods are compared with other known Gammarus species from China.