Venoms are produced by a wide variety of species including spiders, scorpions, reptiles, cnidarians, and fish for the purpose of harming or incapacitating predators or prey. While some venoms are of relatively simple composition, many contain hundreds to thousands of individual components with distinct pharmacological activity. Pain-inducing or “algesic” venom compounds have proven invaluable to our understanding of how physiological nociceptive neural networks operate. In this review, we present an overview of some of the diverse nociceptive pathways that can be modulated by specific venom components to evoke pain. Venoms can be defined broadly as toxins secreted by an animal for the purpose of harming or incapacitating predators or prey. Venoms are widely used by spiders, scorpions, reptiles, cnidarians, fish, and by a few mammals and marsupials. While some venoms are of relatively simple composition, many contain hundreds to thousands of individual components with distinct pharmacological activity ,2,3,4. GMost commonly, venoms are delivered to the victim through wounds resulting from encounters with physical defence mechanisms such as claws, teeth, quills, spines and stingers. However, topical delivery of venoms, for example, to the eye has also evolved, in the case of spitting cobras, multiple times . BroadlyConus geographus are able to switch—rapidly, repeatedly and interchangeably—between venom containing high levels of paralytic toxins when faced with a human or predator, or venom containing prey-specific toxins that are mostly inactive at human targets when hunting for a meal [Cupiennius salei, as well as several snakes, are able to adjust the amount of injected venom based on prey size [For example, cone snails such as r a meal . Similarrey size ,11. ThusOverall, algesic and analgesic venom compounds have proved invaluable to our understanding of how our physiological nociceptive neural networks operate. In this review, we present an overview of the nociceptive channels and neuronal pathways that are commonly modulated by specific venom components. Specifically, we highlight those venoms that induce nociceptive responses, and the diversity of mechanisms that these venoms target to evoke nociception.Pain is indisputably a powerful motivator, and a crucial mechanism necessary for survival, as it allows an organism to alter its normal behaviours to protect the site of injury and facilitate proper tissue healing. Pain also encourages learnt avoidance behaviours that serve to prevent future exposure to noxious stimuli . The evoAt the anatomical level, pain is typically the result of activation of nociceptive or “pain-sensing” peripheral nerve endings, which primarily consist of thinly myelinated Aδ and sparsely or unmyelinated C-fibre neurons . The celV), two-pore domain (K2P) and inward rectifier (Kir) families (voltage-gated and K2Ps), as well as the Na+/K+-ATPase which maintains the required ionic gradients. To a lesser degree, hyperpolarisation-activated cyclic nucleotide-gated, voltage-gated Na+ channels (NaV), and voltage-gated Ca2+ channels (CaV) also contribute to setting the resting membrane potential [V channels leads to the rapid upstroke of the action potential.The threshold of action potential generation, or transformation of a nociceptive signal, is regulated tightly by ion channels involved in setting the resting membrane potential. Of particular importance in this regard are the potassium channels, in particular members of the voltage-gated , activation of NaV) channels are key transmembrane proteins that contribute to setting the resting membrane potential, and that are essential for propagation of action potentials in nociceptive neurons [V1.1–NaV1.9, have been identified and are encoded by the genes SCN1A-SCN5A and SCN8A-SCN11A [Voltage-gated sodium  is of interest as it causes ciguatera, the most common non-bacterial form of fish-borne illness in humans due to the consumption of fish contaminated with ciguatoxins [V activators described below. Studies show that simultaneous activation of all NaV channels by P-CTX-1 produces nocifensive responses when administered subcutaneously or intra-colonically in mice [V1.6 and NaV1.7 activation, as shown by inhibitory pharmacological modulation. In contrast, P-CTX-1 induced visceral pain appears to be predominantly mediated via NaV1.8 [V channels between somatic and visceral innervating nociceptors. In conjunction with these findings, researchers have discovered compounds in painful scorpion venoms that selectively activate NaV1.6 (Cn2) and NaV1.7 (OD1) [Although it is not a venom, the pan-Nauatoxins ,22 Key s in mice . In micea NaV1.8 , highlig.7 (OD1) ,24,25,26.7 (OD1) ,24,25,26V channels are highly conserved across many phyla, the spastic paralysis induced by envenomation with NaV activators has likely contributed to the evolutionary success of these compounds, resulting in convergent recruitment of this pharmacology.As NaV activators also typically elicit nocifensive responses after local injection. While subtype-selectivity for mammalian NaV isoforms is likely not required as activation of at least NaV1.1, NaV1.6, NaV1.7 and NaV1.8 results in pain, structural similarities of mammalian NaV isoforms to prey channels  in conjunction with differences between mammalian isoforms has led to the evolution of highly subtype-selective NaV probes. Accordingly, NaV channel activator toxins have been found in many venomous animals, including cone snails (δ-conotoxin SuVIA from Conus suturatus), spiders , sea anemone , scorpions , wasps , and snakes   see  [27,28,2tivation ,32,33. TV1.1 modulator that impairs channel inactivation, revealed an unexpected role for NaV1.1 in mechanical pain [V1.1 could potentially play a role in the transmission and transduction of mechanical stimuli [V1.1 led to a decrease in nocifensive responses and decreased mechanical sensitisation [The recent discovery of δ-theraphotoxin-Hm1a (Hma1), a selective Nacal pain . Hm1a incal pain . The gen stimuli . Indeed,tisation .V1.1/NaV1.3 antagonist ICA-121431 [V1.1 may be essential for the development and maintenance of chronic visceral pain conditions [V1.1 may be a future target for the treatment of disorders accompanied by chronic visceral pain originating from the colon.Mechanical allodynia and hyperalgesia to colonic distension are crucial contributors to abdominal pain in patients with irritable bowel syndrome (IBS) ,35. ApplA-121431 . FurtherA-121431 . Importanditions . As suchV1.8 [While systematic studies into the co-evolution of the pain system and pain-causing toxins are largely lacking, this evolutionary arms race is poignantly illustrated by the evolution and function of ion channels in specific species that frequent the habitat of venomous animals. For example, Grasshopper mice are resistant to the painful venom of bark scorpion which they prey on, due to a mutation in NaV1.8 . RecentlV1.8 , providiV channels have been extensively investigated for their potential to treat several forms of pain, and, although NaV antagonists are outside the scope of this review, several excellent reviews of venom-derived NaV antagonists are available (see [Venom compounds that modulate Nable (see ).The transient receptor potential (TRP) channels belong to a superfamily of cation channels with diverse tissue distribution in multicellular organisms , and in 2+, Na+, Mg2+ or Zn2+ ions, resulting in action potential generation in several classes of nociceptors. This mechanism is exploited by several venom-derived peptides acting at TRPV1, including RhTx from the Chinese red-headed centipede (Scolopendra subspinipes mutilans) [Mesobuthus martensii [Ornithoctonus huwena [Echis coloratus [Psalmopoeus cambridgei [TRPV1 is activated by capsaicin, the active ingredient in hot chilli peppers, by noxious chemicals, and moderate heat under normal conditions. Interestingly, during in vitro pH conditions similar to that of algesic tissue acidosis, there is a shift in the activation threshold of TRPV1 to room temperature ,51,52. Autilans) , BmP01 fartensii , the sels huwena , venom coloratus , as wellmbridgei   channels, perhaps suggesting that activity at TRPV1 is a natural extension of the pharmacology of venom peptides. Indeed, the vanillotoxins are closely related to K+ channel inhibitor toxins and both VaTx1 (τ/κ-theraphotoxin-Pc1a) and VaTx2 (τ-theraphotoxin-Pc1b) are known inhibitors of KV2.1 channels; this dual activity may contribute to pain-inducing effects in vivo [Interestingly, TRPV1 has relatively high structural similarity to the Shaker family of K in vivo .V2.1, and no inhibition at KV1.2 was observed for any of the vanillotoxins, activity at other K+ channel isoforms cannot be ruled out at present [90 concentration (20 µM) caused TRPV1-mediated paw licking behaviours that were entirely abolished in TRPV1−/− animals [V and TRPV1 channels, suggesting that activity at TRPV1 is not simply a case of molecular and evolutionary pharmacology promiscuity [While VaTx3 (τ-theraphotoxin-Pc1c) appears selective for TRPV1 over K present . Nonethe animals . Interesmiscuity . Recentlmiscuity .Venom peptides have also been crucial in the determination of the structure of TRPV1 and its distinct conformations in activate states ,60,61. MTRPA1 initiates pain signalling due to inflammatory mediators and noxious cold, and has a well-established role in toxin-induced pain . InteresThrixopelma pruriens) is analgesic in certain conditions, and a potent antagonist of several Nav isoforms and TRPA1, possibly through interaction with the extracellular loops of the S1–S4 gating domain of both channel families [Phoneutria nigriventer) is a CaV and TRPA1 antagonist, and has been shown to attenuate mechanical and cold hyperalgesia induced by the TRPA1 agonist allyl isothiocyanate (AITC) in vivo [Crotalus durissus terrificus), is a partial activator of TRPA1 and causes strong desensitisation of the channel, which may be responsible for its analgesic effect in vivo [The spider peptide ProTx-I  [V channels, KV channels consist of four pore-forming α-subunits containing six transmembrane segments (S1–S6) that form a functional tetramer [V1 or KV7) can form heteromultimers with other α-subunits, this results in considerable channel diversity with differing pharmacological and biophysical characteristics [V isoforms that repolarise the membrane potential following an action potential and are pathologically involved in hyperexcitability following injury and neuropathy [In the human genome, forty known genes encode the α subunits of voltage-gated potassium channels (KV) . Similartetramer ,71. As seristics . Sensoryuropathy ,74,75.V channels is to oppose membrane depolarisation or action potential generation and subsequent propagation [V channels have an inhibitory role in sensory neuron excitability [V activity is a hallmark of the hyper-excitability demonstrated in traumatic injuries and neuropathy [The intrinsic function of Kpagation . Thus, Ktability . Decreasuropathy .V toxins, venom peptides can block KV function through occlusion of the pore, or by binding to the voltage-sensing domains to alter channel function [V inhibitors have been isolated from the venom of many species, including cone snails , scorpions , spiders , snakes  and sea anemone , the pae matter ,75,81,82nd KV8.1 . While tth TRPM8 . Similarnjection . In addiThe Acid-sensing ion channel (ASIC) family contains six subunits  encoded by four genes (ASIC1–4) ,86. ASICMicrurus tener tener [−/− mice, despite the injected concentration being orders of magnitude greater than the in vitro EC50 at ASIC1-containing channels (9.4–23.0 nM) [Many venoms are acidic, and it is thus not surprising that acid-sensitive channels such as ASICs might contribute to venom-induced pain either through proton-mediated activation or indirectly as a result of inflammation-induced tissue acidification. However, despite the prominence of ASIC channels in pain pathways, only one venom component that directly activates ASICs has been described so far—MitTx, a 20.8 kDa heteromeric toxin complex from the Texas coral snake er tener . MitTx ier tener . Indeed,23.0 nM) . While MHeloderma suspectum, Heloderma horridum horridum and Apis mellifera; and type 4, which consists of 40–80 amino acid residue peptides that are secreted by some cone snails.Arachnid, insect, cnidarians, and snake venoms are particularly rich sources of phospholipase A2 (PLA2), an enzyme that catalyses the hydrolysis of phospholipids to produce arachidonic acid and lysophosphatidic acid ,91. ConvV channels, thus enhancing the duration of presynaptic membrane depolarisation [Envenomation with snake venom PLA2s can have a range of biological effects, most notably neuromuscular toxicity, myotoxicity, cardiotoxicity, haemolytic and anti-coagulant effects, as well as organ and tissue damage . Extracerisation ,97,98. TApis mellifera, is commonly described as a toxin mediating its effects via activation of PLA2. Indeed, melittin induces hyperactivity of PLA2, possibly as a result of Ca2+ influx [V1.8 and NaV1.9, key ion channels involved in propagation of action potentials in sensory neurons [For instance, melittin, a major pain-producing component from the venom of the honey bee + influx . Melitti+ influx . In addi neurons .Pore-forming peptides are typically produced by bacteria, but are also found in many types of venoms, including cnidarians, ants, spiders, bees and wasps. These toxins are frequently cytolytic due to their ability to disrupt cellular membranes . These pMyrmecia pilosula [2+ ions in vitro. Accordingly, local intraplantar injection of the toxin causes spontaneous pain as well as mechanical allodynia [Direct nociceptive effects have been shown for δ-myrtoxin-Mp1a, an α-helical peptide from the venom of the Jack Jumper ant pilosula . Activitllodynia .Staphylococcus aureus [Staphylococcus aureus and is a known pore-forming toxin responsible for tissue damage, inflammation and bacterial migration. Insertion of α-haemolysin into cell membranes leads to formation of heptameric pores that allow cation influx, including in sensory neurons [A similar mechanism also contributes to the pain-inducing effects of α-haemolysin, a pore forming toxin produced by s aureus . α-Haemo neurons . As a re neurons . In prinStudies focusing on the venoms produced by a myriad of species have provided crucial insight into prey–predator interactions. By isolating the thousands of different peptide components of these venoms, the field has provided new insights into venom-targeted channels and receptors. By taking advantage of millions of years of evolution, researchers have gained highly valuable insights into sensory function and pain signalling in physiological and pathological conditions. Finally, these venom-derived compounds have allowed elucidation of ion channel gating and activation mechanisms, and even species evolution of ion channels and their function.


Mitogen-activated protein kinases (MAP kinases) are a family of kinases that regulates a range of biological processes implicated in the response to growth factors like latelet-derived growth factor (PDGF), epidermal growth factor (EGF), vascular endothelial growth factor (VEGF), and stress, such as ultraviolet irradiation, heat shock, and osmotic shock. The MAP kinase family consists of four major subfamilies of related proteins (extracellular regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), p38, and extracellular regulated kinase 5 (ERK5)) and regulates numerous cellular activities, such as apoptosis, gene expression, mitosis, differentiation, and immune responses. The deregulation of these kinases is shown to be involved in human diseases, such as cancer, immune diseases, inflammation, and neurodegenerative disorders. The awareness of the therapeutic potential of the inhibition of MAP kinases led to a thorough search for small-molecule inhibitors. Here, we discuss some of the most well-known MAP kinase inhibitors and their use in cancer research. Protein kinases are a family of enzymes that phosphorylate proteins on serine, threonine, or tyrosine. Protein phosphorylation brings about changes of their functions, such as their interaction with other proteins, localization, or enzymatic activity. Our genome contains more than 500 protein kinase genes as well as some pseudogenes. Protein phosphorylation plays a critical role in the regulation of numerous cellular properties such as proliferation, differentiation, apoptosis, migration, and adhesion. Therefore, the wrong kinase activity can result in exceptional alterations of these processes. In fact, defective kinases are often found to be oncogenic and can be important for the existence of cancer cells. In addition, the phosphorylation of some proteins, such as protein kinase B, also known as Akt (Akt) ,2, epideSP600125 is a selective and reversible inhibitor of JNK kinases, which has an IC50 for JNK1, JNK2 = 40 nM, and JNK3 = 90 nM. It induces cell death selectively in undifferentiated thyroid cancer cell lines , reducesAS601245 is a cell-permeable JNK inhibitor  inhibits cancer stem cells in vitro and in vivo [D-JNKI-1 (IC50 for JNKs is 2.31 μM) reduces tumor growth in a mouse skin cancer model [BI-78D3 (IC50 for JNKs is 280 nM) sensitizes osteosarcoma to doxorubicin [Several other JNK inhibitors show promise in cancer cells:  in vivo , D-JNKI-er model , and BI-orubicin .SCIO-469  is a selective p38 inhibitor. The IC50 for p38α = 9 nM and for p38β = 90 nM. SCIO-469 enhances bortezomib-induced cytotoxicity against multiple myeloma cells [ma cells , enhancema cells , and decma cells ,54. In ama cells . This inma cells , myelodyma cells , and acuma cells .BIRB-796 (Doramapimod) is a p38 inhibitor  is a selective p38 inhibitor. The IC50 for p38α = 5.3 nM and for p38β = 3.2 nM. LY2228820 enhances bortezomib-induced cytotoxicity and inhibits osteoclastogenesis in multiple myeloma [ myeloma , produce myeloma , and inh myeloma . A Phase myeloma . Althoug myeloma . VX-745 (IC50 for p38α = 10 nM and for p38β = 220 nM) inhibits multiple myeloma cell growth [PH-797804 (IC50 for p38α = 26 nM and for p38β = 102 nM) reduces tumor growth in colon tumor xenografts [A couple other p38 inhibitors show promise in cancer cells: l growth  and PH-7nografts . FR180204 is a selective ERK1/2 inhibitor (nhibitor . The IC5XMD8-92 is a selective ERK5 inhibitor (nhibitor , which hSI113 is a selective SGK1 inhibitor, which has an IC50 for SGK1 = 600 nM. This inhibitor induces autophagy, apoptosis, and cell viability through endoplasmic reticulum stress in endometrial cancer cells [er cells , induceser cells , blocks er cells  (Table 1In the past couple of decades, developments in the small-molecule MAP kinase inhibitor field have led to quite a number of marketed products with a diverse range of the inhibited targets. In addition, many more are still in development and/or improvement. The assortment of these inhibitors allows researchers to choose the most effective and appropriate methodologies suitable for the specific experiments. Preclinical studies both in cell lines as well as in proper animal models provide essential information for the design of clinical studies evaluating the improved efficiency of these agents. Clinical studies with MAP kinase inhibitors ought to determine which MAP kinase inhibitors are most effective for anticancer therapy. Many types of cancers can be targeted by inhibitors, as can be seen in this review. The particular use of MAP kinase inhibitors greatly depends on the genetic background and the precise signaling pathways that direct the cancerogenous properties of the cells in a given cancer type. D-JNKI-1 is a good example showing that peptide inhibitors can go as far as clinical trials [Although there has been significant progress lately in the development of MAP kinase inhibitors, there remains much to be improved. Firstly, for the treatment of cancers, kinase inhibitors have to have high specificity in order to avoid off-target inhibition. However, keeping in mind that there are more than 510 kinases in our cells, it is not an easy task. Sequence similarities between kinases and especially between isoforms of the same family could make it difficult to design specific inhibitors. Peptide inhibitors or aptamers  could bel trials . On the l trials . Therefol trials ,88. Strul trials ,90,91. BInterestingly, there are several alternative compounds which show some potential as MAP kinase inhibitors in cancer research. The natural compound Sulforaphane, extracted from cruciferous vegetables, suppresses ERK and AKT phosphorylation and induces apoptosis through G2/M phase arrest in osteosarcoma cells . MetformIn conclusion, the future seems to be bright for MAP kinase inhibitors and they definitely should not be dismissed.


Hydrolyzed whey-protein  and a saline control  were infused intraduodenally for 60 min in 10 younger  and 10 older  healthy men in a randomized, double-blind fashion. Plasma insulin, glucagon, gastric inhibitory peptide (GIP), glucagon-like peptide-1 (GLP-1), peptide tyrosine-tyrosine (PYY), and amino acid concentrations, but not blood glucose, increased, while ghrelin decreased during the whey-protein infusions. Plasma GIP concentrations were greater in older than younger men. Energy intake correlated positively with plasma ghrelin and negatively with insulin, glucagon, GIP, GLP-1, PYY, and amino acids concentrations (p < 0.05). In conclusion, intraduodenal whey-protein infusions resulted in increased GIP and comparable ghrelin, insulin, glucagon, GIP, GLP-1, PYY, and amino acid responses in healthy older and younger men, which correlated to subsequent energy intake.Protein-rich supplements are used widely for the prevention and management of malnutrition in older people. We have reported that healthy older, compared to younger, adults have less suppression of energy intake by whey-protein—effects on appetite-related hormones are unknown. The objective was to determine the effects of intraduodenally administered whey-protein on glucose, gut hormone, and amino acid concentrations, and their relation to subsequent  A growing awareness of the extent and adverse effects of aging-related muscle loss, including reduced functional capacity and decreased quality of life ,2,3, hasOn the other hand, weight loss and under-nutrition are common in older adults, often associated with, and/or caused by, reduced appetite and food intake. These changes have been termed the ‘anorexia of aging’ ,3, and mPotential mechanisms involved in the regulation of energy intake include variations in gut hormone release and action , glucagon-like polypeptide-1 (GLP-1), and peptide tyrosine tyrosine (PYY)), as well as plasma amino acid concentrations ,15,16. WHealthy aging is associated with a reduced responsiveness to the suppressive effects of nutrients, including protein, on appetite and energy intake ,29. Consad libitum energy intake, in older and younger men. We hypothesized that intraduodenally-administered whey protein would result in load-related responses, related to subsequent energy intake, of glucose, gut hormones, and amino acids, and that these responses would be greater in older than younger men.The aim of the study was to further characterize the effects of intraduodenal whey-protein loads on blood glucose, plasma insulin, glucagon, ghrelin, GIP, GLP-1, PYY, and amino acid concentrations, and their relationships with subsequent www.anzctr.org.au, ACTRN12612000906853). All subjects provided written informed consent prior to their inclusion in the study.The pilot study included 10 healthy young men and 10 healthy older men. Our original study characterized the effect of aging on energy intake, perceptions of appetite and gastrointestinal symptoms, and antropyloroduodenal motility in response to infusion of hydrolyzed whey protein directly into the duodenum (thereby bypassing orosensory and gastric factors) at loads lower than , similar to , and at the upper end  of normal gastric emptying rates , and a saline control, at a rate of 4 mL/min for 60 min (previously published ). We havEach subject was studied on 4 occasions, separated by 7–14 days, in randomized order. The protocol was described in detail previously .Subjects were provided with a standardized evening meal , ~591 kcal) to consume on the night before each study, and were instructed to fast overnight from solids and liquids and to refrain from strenuous physical activity until they attended the laboratory at the University of Adelaide, Discipline of Medicine, Royal Adelaide Hospital, at ~08:30 a.m. On arrival, a small-diameter (3.5 mm) catheter  was inserted into the stomach through an anesthetized nostril and allowed to pass into the duodenum by peristalsis. The infusion port of the catheter was located in the proximal small intestine 14.5 cm from the pylorus .The protein solutions were prepared by dissolving whey protein hydrolysate powder  in varying amounts of saline and water to achieve the desired loads  and to ensure that they were iso-osmotic (640–680 mOsmol/L). The infusions were administered at a rate of 4 mL/min (240 mL over 60 min). The amino acid content of the hydrolyzed  whey protein is presented in Immediately before and at 15 min intervals during the intraduodenal infusion, blood samples (an intravenous cannula was positioned intravenously in the right forearm) for measurement of glucose, insulin, glucagon, ghrelin, GIP, GLP-1, PYY, and amino acids were taken . Blood samples were collected into ice-chilled ethylenediaminetetraacetic acid (EDTA)-coated tubes. Plasma was obtained by centrifugation for 15 min at 3200 rpm at 4 °C and stored at −80 °C for further analysis.Blood glucose concentrations (millimoles per liter) were determined immediately after collection by the glucose oxidase method using a portable glucometer .Total plasma insulin concentration (milliunits per liter) was measured by Enzyme-linked immunosorbent assay (ELISA) immunoassay . The minimum detectable limit was 1.0 mU/L. Intra- and inter-assay coefficients of variation were 3.1 and 9.4%. Homeostatic model assessment (HOMA) index at baseline was calculated according to the following formula: insulin concentration (microunits per liter) × glucose concentration (nanomoles per liter)/22.5 .Total plasma glucagon concentration (picograms per milliliter) was measured by RIA . The minimum detectable limit was 20 pg/mL. The intra- and inter-assay coefficients of variation were 4.4 and 6.3%. The ratio of insulin to glucagon was calculated for each time point in each subject .Plasma total ghrelin concentration (picograms per milliliter) was measured using a radioimmunoassay (RIA) with modifications to the previously published method . The radTotal plasma GIP concentration (picomoles per liter) was measured by RIA with modifications to a previously published method . The staTotal plasma GLP-1 concentration (picomoles per liter) was measured by RIA . The detection limit was 3 pmol/L. Intra- and inter-assay coefficients of variance were 6.4 and 9.5%.4, 10 mM EDTA, 2 g/L gelatin, 0.1 g/L Na-Azide, pH = 7.4) and a 1/12,000 dilution of antiserum for 24 h. The standards and samples were further incubated with 10,000 counts per minute tracer ) for 24 h. Separation of the antibody-bound tracer from free tracer was by second antibody precipitation , and 50 µL of normal rabbit serum diluted 1/50 in wash buffer), incubated 2 h at room temperature then spun at 4000 rpm at 4 °C for at least 20 min, with supernatants poured off and pellets counted in a gamma counter. The detection limit was 1.5 pmol/L. Intra- and inter-assay coefficients of variations were 8.4 and 13.7%.Plasma total PYY concentration (picomoles per liter) was measured using RIA  against human peptide YY 1–36)  and raised in rabbits ,36. This (Sigma-AN-hydroxysuccimimidyl carbamate (AQC) performed at the Australian Proteome Analysis’s Facility established under the Australian Government’s National Collaborative Research Infrastructure Strategy (NCRIS). The maximum increase in plasma amino acid concentration of the highest protein load  at 60 min was calculated as a percentage of the average baseline concentration. The derivatives were separated and quantified by reversed-phase high-performance liquid chromatography (HPLC). The amino acids (with the exception of tryptophan) were detected by fluorescence, whereas tryptophan required UV detection. Before derivatization, 100 μL of plasma samples were diluted 1:1 with internal standard solution  and deproteinized by ultra-filtration through a membrane with 10 kDa nominal molecular weight cutoff . Amino acids contained in the filtrate (100 μL) were labeled using the Waters AccQTag™ chemistry and analyzed using a Waters Acquity™ UPLC system . Histidine data are not available for one of the younger men.Plasma free amino acid concentrations (mmol/L) of asparagine, aspartic acid, alanine, arginine, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine were analyzed using precolumn derivatization with 6-aminoquinolyl-Statistical analyses were performed using SPSS software , 15 min after starting the infusion, immediately before the meal (60 min), and net area under the curve , were determined using a repeated-measures mixed-effect model, with protein load as the within-subject factor and age as the between-subject factor, including baseline values at each treatment visit as a covariate. Post-hoc comparisons, adjusted for multiple comparisons using Bonferroni’s correction, were performed when there were significant main or interaction effects. Within-subject correlations between energy intake (published previously ) and glup < 0.05. Data are presented as mean ± SEM unless otherwise stated.The original study  was powe2) and older men (age: 74 ± 4 years (68–81 years), body weight: 79 ± 7 kg (66–92 kg), height: 1.74 ± 0.05 m, BMI: 26 ± 2 kg/m2). The older men had a lower height and, accordingly, higher BMI than the younger men (p < 0.05). Fasting concentrations of blood glucose, plasma insulin, glucagon, ratio of insulin to glucagon, HOMA-IR, ghrelin, GIP, GLP-1, PYY, total amino acids, and essential amino acids were comparable in healthy younger and older men  and tryptophan  were higher, and of aspartic acid , glutamic acid , and serine  were lower, in older than younger men. The study protocol was well tolerated by all subjects.Body weight did not differ significantly between the younger (mean ± SD (range): age 23 ± 4 years (19–29 years), body weight: 73 ± 7 kg (62–87 kg), height: 1.82 ± 0.02 m, BMI: 22 ± 2 kg/mp = 0.031); glucose was lower after the 90 kcal  and 180 kcal (p = 0.037) protein infusions compared to control in younger, but not older men (p > 0.05), and higher in older than younger men after the 90 kcal protein infusion (p = 0.001). 15 min and AUC blood glucose concentrations were not affected by protein-load or age, with no protein-load by age ; insulin was higher during all protein infusions compared to control . 60 min (p < 0.05) and AUC (p < 0.05) plasma insulin concentrations protein-load dependently increased; concentrations were higher during the 90 kcal and 180 kcal protein infusions compared to control .Plasma insulin concentrations at 15 min protein-load dependently increased ; glucagon was lower in older than younger men during the 90 kcal  and 180 kcal (p = 0.008) protein infusions, higher during all protein infusions compared to control in younger men (p < 0.05), and higher during the 30 kcal protein infusion compared to control in older men (p = 0.003). 60 min (p < 0.001) and AUC (p < 0.001) plasma glucagon concentrations protein-load dependently increased in younger and older men; glucagon was higher during all protein infusions compared to control in younger men , and during the 90 kcal and 180 kcal protein infusions compared to control in older men (p < 0.001).Plasma glucagon concentrations at 15 min protein-load dependently increased ; the ratio was higher during all protein infusions compared to control . The 60 min (p < 0.001) and AUC (p < 0.001) ratio of plasma insulin to glucagon concentrations protein-load dependently increased; ratios were higher during the 90 kcal  and 180 kcal (p = 0.05) protein infusions compared to control.The ratio of plasma insulin to glucagon concentrations at 15 min protein-load dependently increased ; 60 min and AUC plasma ghrelin concentrations were lower during the 90 kcal  and 180 kcal  protein infusions compared to control.15 min, 60 min and AUC plasma ghrelin concentrations protein-load dependently decreased ; 60 min plasma GIP concentrations were higher in older than younger men during the 90 kcal  and 180 kcal (p = 0.013) protein infusions; and 15 min, 60 min, and AUC plasma GIP concentrations were higher during all protein infusions compared to control .15 min, 60 min, and AUC plasma GIP concentrations protein-load dependently increased , and 60 min and AUC plasma GLP-1 concentrations were higher during the 90 kcal  and 180 kcal  protein infusions compared to control.15 min, 60 min, and AUC plasma GLP-1 concentrations protein-load dependently increased ; PYY was higher during the 90 kcal protein infusion compared to control . 60 min (p = 0.018) and AUC (p = 0.042) plasma PYY concentrations protein-load dependently increased in younger and older men.Plasma PYY concentrations at 15 min protein-load dependently increased ; plasma AUC total  and essential (p = 0.014) amino acid concentrations were lower during the 180 kcal protein infusion in older than younger men.15 min, 60 min, and AUC plasma total amino acid concentrations protein-load dependently increased during the infusions ; i.e., the greater the increase in plasma GIP concentrations, the greater the increase in GLP-1. Ghrelin was, within subjects, inversely related to insulin ; i.e., the greater the increase in plasma insulin concentrations, the greater the inhibition of ghrelin production.GIP was, within subjects, related to GLP-1 , which were comparable to , and at the upper end  of normal gastric energy emptying rates , dose-dead libitum meal. There was a dose-dependent suppression of energy intake by protein infusion  in the younger men, and this was greater than the suppression in older men, which was only significant after the 180 kcal protein infusion [ad libitum meal , particeviously ,44,45, aWhile the younger and older men had comparable glucose, insulin, and glucagon concentrations during the 60 min protein infusions, as assessed by the area under the curve method, insulin and glucagon concentrations increased more slowly in older than younger men  and glucose concentrations immediately before the buffet meal (60 min) were lower during the ‘higher’ protein-load infusions 90 and 180 kcal) compared to control in young, but not older, men. We have reported that blood glucose and insulin concentrations were higher during intraduodenal glucose infusions in healthy older than younger men [0 kcal coHealthy older and younger men had comparable AUC ghrelin concentrations during the protein infusions, consistent with responses to mixed-nutrient intake in some ,26 but nPlasma GIP concentrations increased rapidly during the protein infusions and these were higher in older than younger men, which may be related to differences in small intestinal transit of the whey protein. Previously, orally ingested ,20, but GLP-1 and PYY are mainly secreted more distally in the gastrointestinal tract  than GIP (expressed mainly in the duodenum and jejunum), and their plasma concentrations increased more slowly than GIP concentrations during the protein infusions. This was particularly so for PYY concentrations, which increased during the 180 kcal protein load largely after 30 min onwards and more convincingly in the older men. Nevertheless older and younger men had comparable plasma AUC GLP-1 and PYY concentrations during the protein infusions, consistent with responses during intraduodenal infusions of lipid and glucose . Oral glThe older men had a slower increase in plasma concentrations of essential amino acids (lower concentrations at 15 min after starting the whey-protein infusions), particularly leucine, isoleucine, and lysine, than the young men. Our findings are consistent with previous reports that plasma amino acid concentrations peak later and remain elevated longer after amino acid ingestion in older than younger adults, resulting in comparable AUC concentrations between age groups . The infPlasma amino acids concentrations correlated positively with plasma concentrations of insulin, glucagon, GIP, GLP-1, and PYY and negatively with those of ghrelin. Interaction of dietary amino acids, oligopeptides, and proteins in the gut induce the so-called ileal-brake mechanism, including inhibition of proximal gastrointestinal motility, which stimulates the vagus nerve afferents to convey information to the nucleus of the solitary tract in the brainstem and thereby restricts food intake in the short term .p = 0.02) [After their oral ingestion, nutrients empty from the stomach at a rate of 1–4 kcal/min . Gastric = 0.02) . In the  = 0.02) . These f = 0.02) , withoutThis study has several limitations, including the relatively small subject numbers. We studied only men, as they appear to have the greatest ability to respond to energy manipulation , so the ad libitum energy intake. Plasma concentrations of insulin, glucagon, GLP-1, and PYY were increased and ghrelin decreased to a comparable degree by the infusions in both age groups, while GIP responses were greater in older than younger men.Intraduodenal whey-protein infusions resulted in load-dependent changes in gut hormones and amino acids in younger and older men, and these responses were related to subsequent


In about 30% of the cases evaluated by AS, we found AVA ≤ 1.0cm2, indicative of severe AS, with a mean gradient < 40 mmHg,suggestive of moderate AS.4 Thisdissociation makes it difficult to establish an adequate and definitive diagnosis to thepatient with AS, fundamental point in the therapeutic decision making. If, on the onehand, patients with moderate AS do not benefit from valve intervention, those withsevere AS require surgical aortic valve replacement or a transcatheter aorticbioprosthesis implant, especially if they are symptomatic.1-3Severe aortic stenosis (AS) is defined as a significant reduction of the aortic valvearea . These patients present apathophysiology similar to that of diastolic heart failure, with hypertrophy and leftventricular compliance reduction, leading to a "low-flow" state, defined by an ejectedvolume (stroke volume) of < 35 ml/m2 (stroke volume = Diastolic Volume -Systolic Volume / Body Surface).5-7In 2007, Hachicha et al.,5, corroborated by some subsequent studies,8-11 was the demonstration of a better survival ofsymptomatic patients with paradoxical AS after valve intervention when compared toclinical treatment. However, patients with paradoxical AS, despite being benefited byvalve intervention, present higher surgical mortality when compared with patients withclassic AS (mean gradient > 40mmHg).1-3,8,9,11Another important contribution of Hachicha et alIn this paper, we propose an algorithm to facilitate the diagnostic confirmation ofparadoxical AS. In three steps, we perform the Recognition of Paradoxical AS,Measurement Error Evaluation and Pathophysiological Confirmation :1-3 For this reason, patientswith AVA ≤ 1.0 cm2, jet velocity < 4 m/s or meangradient < 40 mmHg and ejection fraction > 50% should be classified ashaving paradoxical AS or low-flow, low-gradient AS with preserved ejectionfraction.1. Recognition of Paradoxical AS: this step is the first and most important.The delay in the diagnosis of paradoxical AS causes delayed intervention,leading to an increase in mortality. The classification of "moderate tosevere" or even "moderately-severe" valvulopathy is not described in any ofthe current guidelines and impairs clinical reasoning.7 3Dechocardiogram is a promising test for more accurate evaluation of VSVE andAVA by planimetry, however, specific studies for the population withparadoxical AS are necessary for its routine indication. Two points areextremely important for the clinical cardiologist. First, in patients withsmall corporeal surface, a reduced AVA may correspond to moderate AS. Inthis way we must always index AVA by the corporeal surface (iAVA), beingthat an iAVA ≤ 0.6 cm2/m2 suggests importantAS. In obese patients (BMI ≥ 30 kg/m2) we must assume alower cut-off value (< 0.5 cm2/m2) so as not tooverestimate the anatomical severity.12 The second data that should be evaluated issystolic blood pressure in gradient measurement moment, which should be lessthan 140 mmHg.1 Higherpressures contribute to underestimating the mean gradient and generate anincrease in the valvulo-arterial impedance, a measure that estimates theventricular afterload added to arterial and valvular overload ventricle, andit is also associated with mortality.13 In summary, the clinical cardiologist shouldremember to index the AVA and make sure that the systolic blood pressure was< 140 mmHg at the time of gradient measurement, while theechocardiographer should be attentive to errors in gradient measurement andmeasure the AVA by the planimetry.2. Evaluation of Measurement Errors: In this stage, we must identify eventualmeasurement errors that justify an underestimated gradient or AVA. Theechocardiographer should be aware of the correct alignment of the Dopplercontinuous wave for velocity and gradient measurement, avoidingunderestimating these measurements. Another orientation is to avoid AVAmeasurement by continuity equation and using whenever possible measurementby planimetry. AVA measurement by continuity equation may underestimate AVA,since such measurement takes into account left ventricular outflow tractarea calculation (VSVE) . The VSVE dimension is usuallymeasured with a 2D echocardiogram, assuming that the VSVE is circular.However, such a structure can often be elliptical, causing measurementerrors.14 However, females maypresent the same anatomic severity as men, but with lower values ofcalcification, being advised to apply differentiated cutoff values forfemale patients (> 1200 AU).15 Pathophysiology of low flow should be confirmed bystroke volume calculation, as previously described. In order to justify lowgradient in a patient with severe AS, he must necessarily present a smallcavity with stroke volume < 35 ml/m2.1-3,5-73. Pathophysiology Confirmation: Finally, we must confirm the pathophysiologyof AS and low-flow, low-gradient. In developed countries, the main etiologyof AS is degenerative, also known as calcific. Valvular calcificationcorrelates with anatomic severity and values greater than 1650 AU, verifiedby computed tomography, suggest anatomically severe AS.Thus, through this 3 steps algorithm, we help in the recognition of paradoxical ASanatomical severity, facilitating the clinician to identify the ideal moment forintervention in this difficult diagnosis entity.


Gastric cancer continues to be the second most frequent cause of cancer deaths worldwide. However, the exact molecular mechanisms are still unclear. Further research to find potential targets for therapy is critical and urgent. In this study, we found that ARPC2 promoted cell proliferation and invasion in the human cancer cell line MKN-28 using a cell total number assay, MTT -2,5-diphenyl-2-H-tetrazolium bromide) assay, cell colony formation assay, migration assay, invasion assay, and wound healing assay. For downstream pathways, CTNND1, EZH2, BCL2L2, CDH2, VIM, and EGFR were upregulated by ARPC2, whereas PTEN, BAK, and CDH1 were downregulated by ARPC2. In a clinical study, we examined the expression of ARPC2 in 110 cases of normal human gastric tissues and 110 cases of human gastric cancer tissues. ARPC2 showed higher expression in gastric cancer tissues than in normal gastric tissues. In the association analysis of 110 gastric cancer tissues, ARPC2 showed significant associations with large tumor size, lymph node invasion, and high tumor stage. In addition, ARPC2-positive patients exhibited lower RFS and OS rates compared with ARPC2-negative patients. We thus identify that ARPC2 plays an aneretic role in human gastric cancer and provided a new target for gastric cancer therapy. Despite the rapid progression in the therapies used to treat gastric cancer patients, it continues to be the second most common cause of cancer deaths worldwide –4. PatieActin-related protein 2/3 complex subunit 2 (ARPC2) is one of the evolutionarily conserved subunits of actin-related protein 2/3 complex (Arp2/3). The other subunits are ARPC1, ARPC3, ARPC4, and ARPC5 and two actin-related proteins Arp2 and Arp3 –9. In thP = 0.009) and a lower OS rate (P = 0.030) than did ARPC2-negative patients. As a result, ARPC2 played a destructive role in human gastric cancer cells and can be used as a potential target for the diagnosis and therapy of gastric cancer.In this article, we determined that ARPC2 promotes both proliferation and invasion in the human gastric cancer cell line MKN-28 by using a cell total number assay, MTT assay, cell colony formation assay, migration assay, invasion assay, and wound healing assay. For downstream genes, we found that cancer-promoting genes were upregulated while tumor suppress genes were downregulated by ARPC2. Moreover, in clinical tissues, ARPC2 was associated with tumor size, lymph node invasion, and tumor stage but not with patients' age, gender, or tumor grade. ARPC2 expressed higher levels in gastric cancer tissues than in normal gastric tissues. Furthermore, ARPC2-positive patients exhibited both a lower RFS rate (2.Human gastric cancer cell MKN-28 was obtained from ATCC (the American Type Culture Collection) . As recommended, MKN-28 was cultured in a humidified incubator at 37°C and 5% COSmall-interfering RNAs used in this article contained two types of ARPC2-siRNA (designated as siARPC2-1 and siARPC2-2 or siAR-1 and siAR-2) and negative control (designated as siNC). They were obtained from GenePharma . Lipofectamin 2000  was used to perform siRNA transfection.We used RT-qPCR to evaluate the mRNA levels of CTNND1, EZH2, BCL2L2, CDH2, VIM, EGFR, PTEN, BAK, and CDH1, which was performed as described in the previous studies . GAPDH wIn this study, the cell total number assay, MTT assay, cell colony formation assay, migration assay, invasion assay, and wound healing assay were carried out to test cell proliferation and invasion. They were all carried out as previously described –15.β-Actin was used as the endogenous control.Western blot analysis was carried out to test the protein level of ARPC2, which was performed as described previously . β-ActinIn total, 110 paraffin-embedded surgical gastric cancer tissue specimens and 110 paraffin-embedded surgical normal gastric tissue specimens were collected at the First Affiliated Hospital of Anhui Medical University  between 2009 and 2015. All experimental protocols were approved by the Ethical Committee of Anhui Medical University and conform to the principles outlined in the Declaration of Helsinki. The pathohistological diagnosis and grade of the patients was based on the World Health Organization grading systems. This study protocol was approved by the institutional review board. Informed consent forms were obtained from all patients. We followed up the patients with gastric cancer, and the average duration was approximately 5 years.Immunohistochemistry analysis was carried out as described earlier . In thist-test. For the experiments involving clinical tissues, the differences were analyzed using Pearson's chi-square test. Patient relapse-free survival (RFS) and overall survival (OS) were analyzed using Kaplan-Meier curves, and the differences were analyzed using the log-rank test. P < 0.05 was considered statistically significant.For the in vitro experiments, the differences were analyzed using unpaired two-tailed Human gastric cancer MKN-28 cells were tranfected with ARPC2-siRNA-1, ARPC2-siRNA-2, or negative control . To further evaluate whether ARPC2 could promote the invasion of MKN-28 cells, migration assay, invasion assay, and wound healing assay were carried out. After transfection with ARPC2-siRNA-1 and ARPC2-siRNA-2, both the migration  and invaIn addition, we carried out large-scale RT-qPCR to screen for genes that were regulated by ARPC2. It has been reported previously that CTNND1, EZH2, BCL2L2, CDH2, VIM, and EGFR have positive correlation to proliferation, invasion, and poor prognosis of gastric cancer. As shown in We collected 110 normal gastric tissues and 110 gastric cancer tissues of archived formalin-fixed paraffin-embedded specimens and detected the protein level of ARPC2 using immunohistochemistry. The positive signal of ARPC2 protein was mainly located in the cytoplasm. As shown in P = 0.001); higher in tumors with lymph node invasion than in tumors without lymph node invasion (P = 0.004); and higher in higher-stage (stages III-IV) tumors than in lower-stage (stages I-II) tumors (P = 0.001). However, there was no significant difference between ARPC2 expression and other clinic-pathological features, including patients' age, gender, and tumor grade  and a lower OS rate (P = 0.030) than did the ARPC2-negative patients.To evaluate the RFS and OS rates of gastric cancer patients with different levels of ARPC2 expression, Kaplan-Meier analyses were performed in the 110 gastric cancer tissues. Every patient was followed up for more than 5 years. As shown in Herein, we confirmed for the first time that ARPC2 had adverse effects in human gastric cancer, and this was the first report on the role of ARPC2 in human cancers. Combining multiple methods that were used in our study, we drew conclusions that ARPC2 promoted both the cell proliferation and invasion of human gastric cancer cells. In clinical samples, the expression level of APRC2 was much higher in gastric cancer tissues than in normal gastric tissues. Furthermore, the expression of ARPC2 was associated with aggressive behaviors of gastric cancer, including large tumor size, lymph node invasion, high tumor stage, and poor prognosis.As for recurrent, metastatic, or advanced gastric cancer, the traditional therapeutic methods, including surgery, chemotherapy, and radiotherapy, showed poor curative effects and patient prognoses , 12, 17.Chen et al. proved that EZH2 promoted the progression and invasion of human gastric cancer [As reported previously, Arp2/3 complex is essential for cell motility , 20. As c cancer , 27. BCLc cancer –30. CDH2c cancer . Vimentic cancer –34. EGFRc cancer –37. Morec cancer –41. BAK c cancer , 43. CHDc cancer , 45. TheIn this study, we reported for the first time the destructive role of ARPC2 in human gastric cancer cells. Both in vitro and clinical studies were performed. A high expression of ARPC2 was associated with both poor RFS and OS rates in gastric cancer patients. Therefore, we propose ARPC2 as a new potential biomarker and therapeutic target for patients with gastric cancer.


Oryza rufipogon Griff.) that enhance salinity tolerance of rice (O. sativa L.), a set of introgression lines (ILs) were developed. The ILs were derived from an O. rufipogon accession collected from Chaling  as the donor, and a widely grown O. sativa indica cultivar 93-11 as the recipient. Through evaluating the salt tolerance of 285 ILs at the seedling stage, a total of 10 quantitative trait loci (QTLs) related to salt tolerance were identified on chromosomes 1, 5, 7 and 9–12, with individual QTLs explaining 2–8% of phenotypic variance. The O. rufipogon-derived alleles at four QTLs improved salt tolerance in the 93-11 background. At the same time, a salt-tolerant IL, 9L136, was identified and characterized. Compared with the recipient parent 93-11, a total of 1,391 differentially expressed genes (DEGs) were detected specifically in 9L136 between salt stress and normal condition through genome-wide expression analysis. Of these, four DEGs located in the QTL regions carried by 9L136, suggesting that the four genes might be candidates associated with salt tolerance. Both the highly salt-tolerant ILs and the favorable O. rufipogon-derived QTLs identified in the present study will provide new genetic resources for improving the resistance of cultivated rice against salinity stress using molecular breeding strategies in the future.Soil salinity is a common abiotic stress affecting crop productivity. To identify favorable alleles from wild rice ( Oryza sativa L.), one of the most important food crops in the world, has medium sensitivity to salinity stress, and shows symptoms of damage and a dramatic reduction in productivity when the soil soluble salt reaches 0.3%  and potassium (K+) content, a total of 11 QTLs were located on chromosomes 1, 4, 6, and 9, which explained 6.4–19.6% of phenotypic variation , encoding a C2H2-type zinc finger transcription factor, was cloned and had a negative effect on drought and salt tolerances through regulating stomatal closure in rice , the wild progenitor of Asian cultivated rice (O. sativa L.), is an important primary gene pool for improving rice. Many favorable genes have been identified and characterized from wild rice  from China, and identified QTLs for salt tolerance. Four candidate genes in QTL regions associated with salt tolerance were identified through integrating RNA sequencing and QTL mapping. The results will aid in utilizing alien alleles to improve tolerance to salt stress in modern rice breeding.Common wild rice  from Chaling  based on an advanced backcross (BC3) and consecutive selfing strategy , floated on water for 4 days, and then transferred to float on Yoshida’s culture solution . About 1Using the cetyltrimethyl ammonium bromide method , DNA of P < 0.01 and LOD ≥ 2. The proportion of observed phenotypic variance explained by a QTL was estimated by the coefficient of determination (R2) from the corresponding linear model fitting.Based on the genotypic analysis of 285 ILs using 142 polymorphic SSR markers, QTLs for salt tolerance were determined using single-point analysis in Map Manager QTXb17 . The staSix physiological characteristics associated with abiotic stress were measured as described previously : contentn = 3) were subsequently sequenced using an Illumina HiSeq2500 Sequencer. After removing adaptor sequences and low-quality reads, the high-quality paired-end reads were mapped to the Nipponbare reference genome (MSU Rice Genome Annotation Project Release 7) using the spliced read mapper TopHat version 2.0.12  and control  using TRizol  and purified using a Qiagen RNeasy kit . Each sample had three biological replicates. The purified RNA was used to construct the cDNA library using a NEBNext Ultra RNA Library Prep kit. The 12 libraries . The cDNA samples were subjected to qRT-PCR quantification with three biological replicates and performed with SYBR Green Master Mix  according to the product manual on a CFX96 Real Time System . Reference genes and specific gene primers are listed in Supplementary Table O. rufipogon-derived favorable alleles associated with salt tolerance, we investigated both STS and survival rate of 285 ILs and the recurrent parent 93-11 under 125 mM NaCl treatment. Phenotypic measurement showed that the frequency distributions of the two traits in the IL population accorded with the normal distribution, and both STS and the survival rate of the recurrent parent 93-11 (1.07 and 43%) were lower than the means for ILs (1.81 and 54%) at the seedling stage . The results indicated that the recurrent parent 93-11 was more susceptible to salt stress, and that the O. rufipogon genome might harbor beneficial alleles associated with enhanced salt tolerance in the cultivated rice background.To identify O. rufipogon were detected, including 1082 homozygous and 276 heterozygous segments . They were located on chromosomes 1, 5, 7 and 9–12, and each QTL explained 2–8% of the phenotypic variation . Additionally, the O. rufipogon-derived alleles at four loci (40% of QTLs) improved salt tolerance. Among these QTLs, six located on chromosomes 1 and 9–12 were detected using STS and survival rate as indexes. For example, qST10 was located near the SSR marker RM258 on chromosome 10, and qST12 near SSR marker RM7102 on chromosome 12. Moreover, alleles from O. rufipogon at the two QTLs could improve salt tolerance at the seedling stage in the 93-11 background. However, the other four QTLs  were only detected by one index.Based on the genotypic analysis of 285 ILs using 142 polymorphic SSR markers, a total of 1358 chromosomal segments from Table 3). Comparison of introgression segments with chromosomal location of QTLs carried by ILs showed that the four lines harbored two positive-effect QTLs for salt tolerance from O. rufipogon  , had the highest STS and survival rate among the four ILs, and its average survival rate was 84%, which increased by 95.3% compared with 93-11 (Table 3). Therefore, 9L136 was chosen for further characterization, as a highly salt-tolerant IL.To select the genetic materials for further breeding salt-tolerant rice varieties and cloning salt tolerance-related genes, we performed six independent-experiment evaluations and identified four ILs , which showed stable salt tolerance in multiple experiments, and had significantly higher STS and survival rates than those of the recurrent parent 93-11 , and 9L136 had a distinctly lower malondialdehyde content in response to 4 days of salt treatment compared with 93-11 . We also investigated the contents of soluble sugars, a class of compatible osmolytes, and observed that this was significantly higher in 9L136 than in 93-11 under salt treatment for 2, 3, and 4 days . Additionally, the proline content in 9L136 was significantly higher than that in 93-11 under salt stress . Overall, the results implied that 9L136 did not suffer from oxidative stress as much as the recurrent parent 93-11 during salt stress, suggesting that favorable alleles from wild rice carried by 9L136 might improve the salt tolerance of rice in the 93-11 background.Generally, salt stress is accompanied by osmotic stress, leading to reactive oxygen species damage and osmolyte accumulation . To inveTo explore candidate genes related to salt tolerance, RNA-seq analysis was performed using RNA extracted from roots of 93-11 and 9L136 after 0 (CK) and 1 h (ST) treatment with 125 mM NaCl. A total of 692 million clean reads . Further comparison of DEGs between 9L136-ST/9L136-CK and 93-11-ST/93-11-CK found that 1,391 genes showed differentially expressed only in 9L136-ST/9L136-CK but not in 93-11-ST/93-11-CK , suggesting that these genes should be specifically induced in IL 9L136 under salt stress.Global expression analysis showed that a total of 4,650 DEGs, including 2,504 up-regulated and 2,146 down-regulated genes, were detected in IL 9L136 between salt stress (9L136-ST) and normal condition (9L136-CK); and a total of 4,291 DEGs, including 2,173 up-regulated and 2,118 down-regulated genes, were detected in the recipient parent 93-11 between salt stress (93-11-ST) and normal condition (93-11-CK) (qST5-2 and qST10) carried by 9L136 harbored a total of 209 genes. To narrow down the candidate genes for salt tolerance, we further compared the chromosomal location between the 1,391 DEGs and QTL regions, and found that four genes, including acetyltransferase (LOC_Os05g31254), calmodulin-related calcium sensor protein , LRP1 (LOC_Os05g32070), and GRAM domain containing protein , were co-localized with the QTLs (qST5-2 and qST10) . Taken together, four candidate genes associated with salt tolerance at the seedling stage were identified through the strategy of combining QTL mapping and genome-wide expression analysis.Additionally, based on the gene annotation information of the rice reference genome (the TIGR Rice Genome Annotation Database), chromosomal regions of two QTLs ; and qST10, detected on chromosome 10 near RM258, was located on a similar region with a QTL cluster  reported by O. rufipogon-derived alleles at these QTLs/QTL cluster improved salt tolerance in the cultivated rice background.In previous studies, many QTLs related to salt tolerance were identified and located on 12 rice chromosomes, with most of them identified in cultivated rice. There are few reports of QTLs for salt tolerance in common wild rice. A total of 15 QTLs for salt tolerance were identified on chromosomes 1, 2, 3, 6, 7, 9, and 10 using a set of ILs derived from Yuanjiang common wild rice, and the wild rice derived alleles at 13 QTLs (86.7%) could enhance salt tolerance in the Teqing background . In our O. sativa populations. The results showed that qST5-1 near RM3328 on chromosome 5 was in a similar location to qDLRa5-2 and QSst5a , calmodulin-related calcium sensor protein , LRP1 (LOC_Os05g32070), and GRAM domain containing protein . Notably, both LOC_Os05g31620  and LOC_Os10g34730  had higher expression level in 9L136-ST than in 93-11-ST. There are many GRAM domain-containing protein genes in rice, but only one GRAM domain-containing protein has been reported in rice and may function in plant development , and the GEO accession number is GSE101734.The original RNA-seq data has been uploaded to NCBI (Manuscript draft: SW, FL, XM, LT, and CS. Experiments: SW, MC, JZ, and WC. Analyzing data: XM. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer MJ and handling Editor declared their shared affiliation, and the handling Editor states that the process nevertheless met the standards of a fair and objective review.


Mild or transient TWEAK/Fn14 activation contributes to tissular repair and regeneration while excessive or persistent TWEAK/Fn14 signals may lead to severe inflammatory infiltration and tissue damage. TWEAK also regulates cell fate of keratinocytes, involving the function of Fn14-TNF receptor-associated factor-TNF receptor axis. By recruiting inflammatory cells, promoting cytokine production, and regulating cell fate, TWEAK/Fn14 activation plays a pivotal role in the pathogenesis of various skin disorders, such as psoriasis, atopic dermatitis, cutaneous vasculitis, human papillomavirus infection and related skin tumors, and cutaneous autoimmune diseases. Therefore, the TWEAK/Fn14 pathway may be a potential target for the development of novel therapeutics for skin inflammatory diseases.Tumor necrosis factor- (TNF-) like weak inducer of apoptosis (TWEAK) participates in multiple biological activities via binding to its sole receptor—fibroblast growth factor-inducible 14 (Fn14). The TWEAK/Fn14 signaling pathway is activated in skin inflammation and modulates the inflammatory responses of keratinocytes by activating nuclear factor- Tumor necrosis factor- (TNF-) like weak inducer of apoptosis (TWEAK) is a member of the TNF ligand superfamily and is initially described as an inducer of apoptosis in transformed cell lines . TWEAK aα, interleukin- (IL-) 1, IL-6, and interferon-γ [Inflammation is one of the basic characteristics of skin disorders, especially the chronic inflammatory diseases that include psoriasis, atopic dermatitis (AD), cutaneous vasculitis, and cutaneous lupus erythematosus . The occrferon-γ , 41, whirferon-γ , 42, 43.In this review, we update recent advances in the function of TWEAK/Fn14 signals in different skin inflammation and also highlight the potential roles of this pathway as therapeutic target in the management of various skin diseases.β-pleated sheet structure that forms a trimeric aggregate. The grooves between the subunits of the trimers serve as binding sites for the receptor [TWEAK is initially synthesized as type II transmembrane proteins of 249 amino acids and can be cleaved by furin into sTWEAK with biologic activities . The C-treceptor . Fn14 isreceptor .The extracellular cysteine-rich domain of Fn14 contains three disulphide bonds. By analyzing the structure of Fn14, it recently revealed a highly conserved core region (Ala34–Ala69) with very few flexible side chains . This reα [Keratinocytes are the major component of the human epidermis. They secrete a broad spectrum of cytokines including proinflammatory cytokines, chemokines, and immunomodulatory cytokines and establish the local cytokine and chemokine milieu, which mediate multiple local and systemic consequences, such as migration of inflammatory cells, activation of immune responses, and proliferation and differentiation of keratinocytes and fibroblasts –50. Keraα , 51, 52.α . KCs havα . In addiα . Therefoα, IL-6, and matrix metalloproteinases [β-induced synthesis and secretion of connective tissue growth factor which promotes collagen synthesis [α, IL-1β, IL-4, IL-17, IL-23, and other cytokines that may trigger or exacerbate inflammatory responses in skin [α, MCP-1, IL-1α, IL-1β, IL-6, and IL-8, which are fundamental in inflammation and angiogenesis [Other types of skin cells are also involved in cutaneous inflammation through secreting cytokines or other components. Dermal fibroblasts are activated under inflammatory condition and produce TNF-teinases . Fibroblynthesis . Paracriynthesis , 59. Inf in skin . Vasculaogenesis .β1 exhibits a synergistic effect on the TWEAK-induced RANTES production by keratinocytes [κBα in keratinocytes [Increasing evidences suggest that TWEAK significantly enhances the synthesis of cytokines in resident cells in skin tissues. TWEAK/Fn14 activation promotes the expression and/or secretion of various cytokines that are involved in inflammatory responses, including IL-6, IL-8, GM-CSF, MCP-1, and RANTES . Especiainocytes . TWEAK ainocytes , 64. Morinocytes . TWEAK cinocytes . Furtherinocytes .β signaling increases collagen production and Fn14 expression in cultured fibroblasts, and overexpressing Fn14 can enhance the expression of extracellular matrix genes in these cells upon TWEAK stimulation [The proinflammatory effects of the TWEAK/Fn14 axis have been described in other cell types. TWEAK induces secretion of prostaglandin E2 (PGE2), IL-6, IL-8, RANTES, and IP-10 in dermal fibroblasts . TGF-β smulation . Moreovemulation , 69, indmulation . Furthermulation . TWEAK umulation . Obviousα stimulation downregulates TNFR1 but upregulates TNFR2 expression [TWEAK regulates cellular responses ranging from proliferation to cell death in a manner highly dependent on the cell type and the microenvironmental context. The biological activities of TWEAK can be mediated through two structurally distinct receptor subtypes: TNFR1 and TNFR2, with the major difference in their cytoplasmic tail. Most cells of the human body express TNFR1, while the expression of TNFR2 is much more restricted . Additiopression . These fκB kinase complex and cellular inhibitor of apoptosis proteins (cIAP) to the TNFR1 signaling complex, thus the necessity for the activation of the classical NF-κB pathway. A complex of TRAF2 with cIAP1, cIAP2, and TRAF1 has further been implicated in the inhibition of TNFR1-induced activation of caspase-8 [κB pathway [The cytoplasmic domain of Fn14 contains a phylogenetically conserved binding motif, and TRAF1, TRAF2, TRAF3, and TRAF5 are able to bind to this site . TNFR1 haspase-8 . Hence,  pathway , 79. TheRecently, we found a switch of TNFR expression profile in keratinocytes under different skin inflammations. In normal keratinocytes, TNFR1 expression is prominent, leading to cell death upon TWEAK stimulation , 64. IntκB and nuclear NF-κB are also elevated, indicating activation of the NF-κB signaling pathway [κB activation in psoriatic inflammation results in the production of multiple proinflammatory cytokines, which further mediate the trafficking and homing of T cells, monocytes, eosinophils, natural killer cells, and mast cells [Psoriasis is one of the most common inflammatory skin diseases. Accumulation of RANTES, IL-8, IP-10, and MCP-1 is prominent in lesional skin of patients with psoriasis . Moreove pathway , 80. NF-st cells , 82. TheκB pathway is abnormally skewed towards the former, resulting in increased keratinocyte survival and epidermal hyperproliferation [κB-activated proliferation marker Ki-67 and antiapoptotic proteins  are expressed at higher levels in keratinocytes upon TWEAK stimulation, further suggesting that TWEAK signals participate in psoriatic inflammation [In psoriasis, the balance between the antiapoptotic and cell cycle inhibitory roles of NF-feration . The NF-ammation , 83. In ammation .α, RANTES, IP-10, and interferon-γ, which are upregulated in the lesional skin of patients with AD [AD is a chronic inflammatory disease characterized by a relapsing form of skin inflammation, disturbance of epidermal barrier function, eczema, and spongiosis formation. Histologically, AD is featured with the infiltration of T cells, macrophages, and dendritic cells. These infiltrating cells are known to secrete a range of cytokines, including IL-8, TNF- with AD .Unlike other inflammatory diseases such as psoriasis or autoimmune diseases , 86, theα mRNA in keratinocytes was observed under TWEAK stimulation, and TNF-α is highly expressed in lesional skin of AD but not in healthy skin [α in the induction of keratinocyte apoptosis, contributing to the formation of AD lesions [α, which binds to TNFR1 and triggers the extrinsic pathway of apoptosis [α [α in the lesional skin may together induce apoptosis of keratinocytes under AD inflammation. Further insight about the function of the TWEAK/Fn14 pathway in AD is expected.Furthermore, a significant increase of TNF-thy skin . Actuall lesions . Previou lesions . TWEAK bpoptosis , 90. Howtosis [α , 89. TheCutaneous vasculitis includes a wide range of diseases that affect the blood vessels of skin and share a common pathological feature of endothelial damage and perivascular leukocyte infiltrates. Recent study showed that TWEAK and Fn14 are significantly expressed in the dermal vessel of lesional skin in patients with urticarial vasculitis but not in healthy controls . MoreoveκBα, phosphorylated ERK1/2, and phosphorylated JNK1/2 and promoted cell proliferation in a dose-dependent manner; anti-human Fn14 mAb can abrogate such effect of TWEAK on human umbilical vein endothelial cells [κB activation and chemokine production in human endothelial cells, thus promoting leukocyte migration and vascular injury in cutaneous vasculitis.Previously, it was found that TWEAK can induce the proliferation and migration of endothelial cells . TWEAK tal cells . TWEAK eal cells . In a hual cells . Moreoveal cells , 95. It al cells . TWEAK aal cells . TherefoκB are highly expressed in these areas. These findings indicate TWEAK/Fn14 activation in HPV16-infected keratinocytes [By interacting with Fn14, TWEAK is an inducer of apoptosis of keratinocytes. However, the role of the TWEAK/Fn14 pathway in regulating the cell fate of HPV-infected keratinocytes is characterized by increased proliferation instead of apoptosis . Fn14 exinocytes . Meanwhiinocytes . E6 has inocytes . E6 can inocytes . Based oMoreover, HPV-infected keratinocytes in anogenital lesions are generally considered to be etiologically associated with carcinogenesis. The HPV16-induced keratinocyte immortalization has been suggested to correlate closely with epidermis-originated malignancies, such as cervical cancer and cutaneous or oropharyngeal squamous cell carcinoma , 99. TheTWEAK/Fn14 activation in autoimmune diseases is strongly supported by a growing number of experimental evidences –105. Theα, CXCL1, and CXCL5) in skin of MRL/lpr mice irradiated with ultraviolet B [Cutaneous lupus erythematosus is characterized by local activation of immune complexes or complement, autoreactive B cells and T cells and overexpression of cytokines and chemokines . The TWEviolet B . These fSystemic sclerosis affects skin tissue by chronic inflammation, vascular injuries, and excessive fibrosis. Recent study suggested that the interaction between sCD163 (marker of monocytes/macrophages) and TWEAK is associated with systemic sclerosis . CD163 iThe TWEAK-Fn14 axis may be also involved in the pathogenesis of polymyositis or dermatomyositis . Serum lκB and extracellular signal-regulated kinase pathways. Interestingly, TWEAK upregulates sheddases such as a disintegrin and metalloproteinase 17, leading to BP180 loss in keratinocytes. Therefore, TWEAK/Fn14 activation may contribute to the pathogenesis of bullous pemphigoid. The actions of TWEAK on target cell or animal models in different skin diseases are listed in Recently, we found that TWEAK/Fn14 signaling plays a pivotal role in the pathogenesis of bullous pemphigoid . The serα and IL-6, in skin lesions [β, and tissue inhibitor of metalloproteinase-1) was observed in these subjects [The TWEAK/Fn14 pathway has become a potential therapeutic target. Growing evidences suggest that TWEAK inhibition can ameliorate inflammatory reaction and tissue damage in several animal models of autoimmune and inflammatory diseases, such as chronic autoimmune arthritis, systemic lupus erythematosus, and experimental autoimmune encephalomyelitis , 117. Th lesions . Anti-TWsubjects .κB activation, and cytokine secretion and exhibits antitumor effect [κB pathway-specific agonistic activity, but do not photocopy other activities of TWEAK [TWEAK/Fn14 signals also show therapeutic potential in the management of tumors. Firstly, agents that inhibit TWEAK binding to Fn14 may have potential therapeutic utility . Anti-TWr effect , 122. Fnr effect . Furtherof TWEAK .TWEAK is a multifunctional cytokine expressed on various cell types and tissues and acts via binding to its sole receptor Fn14. TWEAK/Fn14 activation contributes to various pathological processes, including cell proliferation and death, angiogenesis, carcinogenesis, and inflammation. TWEAK/Fn14 signals are involved in the pathogenesis of multiple skin diseases including inflammatory skin diseases, autoimmune skin diseases, cutaneous vasculitis, HPV infection, and tumors. The involvement of this pathway has made it a promising therapeutic target of braking the never-ending cycle of local inflammation and tissue destruction. In a variety of the disease models, soluble TWEAK- and Fn14-specific antibodies and other drug formats have exhibited promising therapeutic effects.However, the precise mechanism underlying the roles of TWEAK/Fn14 activation in inflammatory and autoimmune diseases, especially in cutaneous diseases, is not fully elucidated. In addition, the therapeutic effects on the inhibition or activation of TWEAK/Fn14 pathway have not been well explained. And based on the preclinical findings, we are supposed to explore the clinical value of TWEAK- or Fn14-related agents.


Salmonella enterica isolated from animals in slaughterhouses before slaughter. A total of 143 non-duplicate Salmonella were recovered from 1,000 fresh fecal swabs collected from four major pig slaughterhouses  and four major chicken slaughterhouses  between March and July 2016. Among Salmonella isolates from pigs, the predominant serovars were Salmonella Rissen  and Typhimurium , and high antimicrobial resistance rates were observed for tetracycline  and ampicillin . Class 1 integrons were detected in 10.2% (5/49) of these isolates and all contained gene cassettes aadA2 (0.65 kb). Two β-lactamase genes were detected among these isolates, and most of these isolates carried blaTEM-1 (46/49), followed by blaOXA-1(4/49). Seven STs  were detected in these isolates, and the predominant type was ST469 (19.6%). Among Salmonella isolates from chickens, the predominant serovars were Salmonella Indiana  and Enteritidis , and high antimicrobial resistance rates were observed for nalidixic acid , ampicillin  and tetracycline . Class 1 integrons were detected in 23 isolates , which contained empty integrons  or gene cassettes drfA17-aadA5 , aadA2 , drfA16-blaPSE-1-aadA2-ereA2  or drfA1-aadA1 . Three β-lactamase genes were detected, and all 94 isolates carried blaTEM-1, followed by blaM-55CTX- (n = 19) and blaSPE−1 (n = 3). Five STs were found in these isolates, and the predominant type was ST17 (71.3%). Our findings indicated that Salmonella was widespread in animals at slaughter and may be transmitted from animal to fork.The current study aimed to analyze the prevalence and characterization of  Salmonella enterica is a major global foodborne pathogen  and four major chicken slaughterhouses with process capacity of 10,000–40,000 chickens per day (100 samples per chicken slaughterhouse). The animal slaughterhouses are respectively located in Weihai, Ciyao, Zhucheng, and Yantai regions in Shandong province, China. Sampling was carried out before slaughter, and at the time animals from different farms has been mixed.Salmonella colonies were identified using both the VITEK system  and polymerase chain reaction (PCR) amplification of the inherent gene invA  and was incubated at 37°C for 16 to 18 h. After that, 0.1 mL of the BPW suspensions was sub-cultured in 10 mL subpackaged Rappaport-Vassiliadis (RV) broth at 42°C for 24 h. One loopful of each RV broth culture was then plated onto xylose lysine tergitol 4 agar plates, and was incubated at 37°C for 24 to 48 h  , ampicillin , cefotaxime , ciprofloxacin , florfenicol , gentamicin , nalidixic acid , spectinomycin , tetracycline , and sulfamethoxazole/trimethoprim . Escherichia coli (ATCC25922) was used as a quality control. The results were interpreted based on the Clinical and Laboratory Standards Institute (CLSI) standards guidelines  isolates.The Kirby-Bauer disk diffusion method was used in this study to examine resistance of es CLSI, . SalmoneBacterial DNA was extracted using a TIANamp bacteria DNA kit  according to the manufacturer's instructions. The gene cassettes within the variable region of class I integrons were detected via polymerase chain reaction (PCR), using previously described primers and procedures , and the alleles and STs were assigned according to the MLST scheme at http://mlst.warwick.ac.uk/mlst/dbs/Senterica. A minimum spanning tree was created using Bionumerics software 6.5 , according to the instructions (the unweighted pair group method of arithmetic averages method).The MLST analysis was performed by sequencing the fragments of seven housekeeping genes . The Salmonella isolates were recovered , including 13 from Weihai , 9 from Ciyao , 11 from Yantai , and 16 from Zhucheng  . The 49 Salmonella belonged to 6 serovars, including Salmonella Rissen (n = 28), Typhimurium (n = 14), Grampian (n = 3), Derby (n = 2), Indiana (n = 1), and Enteritidis (n = 1). The most common serovars were Salmonella Rissen  and Typhimurium  , including 23 from Weihai , 33 from Ciyao , 17 from Yantai , and 21 from Zhucheng  . These 94 Salmonella isolates belonged to 4 serovars, including Salmonella Indiana (n = 67), Enteritidis (n = 23), Typhimurium (n = 3), and Hadar (n = 1). The dominant serovars were Salmonella Indiana  and Enteritidis   and ampicillin 16/49, 32.7%). In addition, 7 isolates  exhibited MDR . In addition, 2 isolates were susceptible to all antibiotics used in this study.All 94 isolates from chicken slaughterhouses were susceptible to amoxicillin/clavulanic acid and sulfamethoxazole/trimethoprim. But most isolates were resistant to nalidixic acid , ampicillin 87/94, 92.6%), and tetracycline . Eighty-six isolates  exhibited MDR . Two β-lactamase genes were detected among the isolates, most of the isolates carried blaSPE-1 (n = 46) and blaOXA-1(n = 4) , including 4 %, includSalmonella Indiana, 5 Enteritidis and 2 Typhimurium. Of these isolates, 5 groups of resistance gene cassettes were detected: empty integrons , drfA17-aadA5 , aadA2 , drfA16-blaPSE-1-aadA2-ereA2 , and drfA1-aadA1 . Three β-lactamase genes were detected among these isolates. Most of the isolates carried blaTEM-1 (n = 94), followed by blaCTX-M-55 (n = 19) and blaSPE-1(n = 3) , including 16 Salmonella isolates were divided into 9 STs, including 7 STs from pigs , and 5 STs from chickens . The STs identified in the present study showed the following correlations with Salmonella serovars: ST11 with Salmonella Enteritidis, ST17 with Indiana, and ST469 with Rissen.One hundred and forty-three Salmonella Typhimurium. ST11 and ST3172 belonged to one clone complex, and had the same serovars of Salmonella Enteritidis.BioNumerics software version 6.5 was used to generate a minimum-spanning tree based on all the sources of STs Figure . The domSalmonella isolation rate from pigs (8.2%) was much lower than that (71.8%) in Jiangsu province, China , which may be associated with the fact that the antimicrobial is frequently used in pig farms in China  was similar to the result reported for frozen chicken meat in Shandong province (26.3%), China , similar with the result conducted in China  was observed in this study, which may be associated with the fact that third-generation cephalosporins have become the primary drugs for the treatment of salmonellosis because of the increase in fluoroquinolone resistance. In addition, the results of the present study showed the high prevalence of multidrug resistant Salmonella isolates in chickens (91.5%), much higher than those reported in Henan province (46.0%), China  from chickens were higher than that (14.3%) from pigs, and the higher occurrence of MDR Salmonella isolates from chickens likely reflects the extensive use of antibiotics during intensive rearing. In addition, MDR Salmonella is serotype-dependent  than Salmonella from pigs (10.2%) (P < 0.05). Class 1 integrons are often associated with MDR Salmonella isolates, consistent with the result of the present study. In addition, the Salmonella isolates carrying class 1 integrons included Salmonella Typhimurium, Enteritidis, and Indiana.In the present study, PCR identified class 1 integrons in 19.6% of Salmonella isolates recovered from pigs and chickens: blaTEM-1, blaPSE-1, blaOXA-1, and blaM-55CTX-. Most isolates carried blaTEM-1, consistent with the report from meat and milk products in Egypt (Ashraf et al., blaOXA-1, followed by blaTEM-1, blaPSE-1, and blaCMY-2 (Li et al., Salmonella from pigs carried blaTEM-1 whereas only 16 were resistant to ampicillin, and only 88 out of 94 Salmonella carrying blaTEM-1 from chickens showed resistant to ampicillin may be associated with the expression status of blaTEM-1 genes and is needed to be further studied.Four β-lactamase genes were detected among blaCMY-2 encodes resistance to third-generation cephalosporins, an important class of antibiotics used to treat complicated cases of salmonellosis (Gonzalez-Sanz et al., blaCMY-2-positive Salmonella in China was low and was only reported in Shanxi and Sichuan (Yang et al., In addition, Salmonella from pigs and chickens. ST19 and ST34 have continually been reported to cause human salmonellosis in recent years, and these bacteria belong to the same serotype, Salmonella Typhimurium (Cai et al., Salmonella Enteritidis, represented by ST11 and ST3172. These findings suggested that serovars and STs were tightly coupled (Sukhnanand et al., Salmonella Grampian, which causes an unusual increase in human cases of Salmonella Grampian infections (Horvath et al., Salmonella could spread from animals to humans via pork and chicken products (Osman et al., The MLST results revealed 9 STs identified in Salmonella isolated from animals in slaughterhouses in Shandong province, China. In addition, this study highlights the necessity to carry out the long-term surveillance for Salmonella recovered from food-producing animals.Collectively, our findings exhibit the prevalence and characteristics of WC and SS: conceived and designed the study. XZ and CY: performed the experiments and analyzed the data. XZ, WC, and SS: wrote and revised the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.