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Activating mutations in MYD88 promote malignant cell growth and survival through hematopoietic cell kinase (HCK)–mediated activation of Bruton tyrosine kinase (BTK). Ibrutinib binds to BTKCys481 and is active in B-cell malignancies driven by mutated MYD88. Mutations in BTKCys481, particularly BTKCys481Ser, are common in patients with acquired ibrutinib resistance. We therefore performed an extensive medicinal chemistry campaign and identified KIN-8194 as a novel dual inhibitor of HCK and BTK. KIN-8194 showed potent and selective in vitro killing of MYD88-mutated lymphoma cells, including ibrutinib-resistant BTKCys481Ser-expressing cells. KIN-8194 demonstrated excellent bioavailability and pharmacokinetic parameters, with good tolerance in rodent models at pharmacologically achievable and active doses. Pharmacodynamic studies showed sustained inhibition of HCK and BTK for 24 hours after single oral administration of KIN-8194 in an MYD88-mutated TMD-8 activated B-cell diffuse large B-cell lymphoma (ABC DLBCL) and BCWM.1 Waldenström macroglobulinemia (WM) xenografted mice with wild-type BTK (BTKWT)– or BTKCys481Ser-expressing tumors. KIN-8194 showed superior survival benefit over ibrutinib in both BTKWT- and BTKCys481Ser-expressing TMD-8 DLBCL xenografted mice, including sustained complete responses of >12 weeks off treatment in mice with BTKWT-expressing TMD-8 tumors. The BCL_2 inhibitor venetoclax enhanced the antitumor activity of KIN-8194 in BTKWT- and BTKCys481Ser-expressing MYD88-mutated lymphoma cells and markedly reduced tumor growth and prolonged survival in mice with BTKCys481Ser-expressing TMD-8 tumors treated with both drugs. The findings highlight the feasibility of targeting HCK, a key driver of mutated MYD88 pro-survival signaling, and provide a framework for the advancement of KIN-8194 for human studies in B-cell malignancies driven by HCK and BTK.

Background Adrenocortical carcinoma (ACC) is an extremely rare, aggressive tumor with few effective therapeutic options or drugs. Mitotane (Mtn), which is the only authorized therapeutic drug, came out in 1970 and is still the only first-line treatment for ACC in spite of serious adverse reaction and a high recurrence rate. Methods By in silico analysis of the ACC dataset in the cancer genome atlas (TCGA), we determined that high expression levels of cyclin-dependent kinase-1 (CDK1) were significantly related to the adverse clinical outcomes of ACC. In vitro and in vivo experiments were performed to evaluate the role of CDK1 in ACC progression through gain and loss of function assays in ACC cells. CDK1 inhibitors were screened to identify potential candidates for the treatment of ACC. RNA sequencing, co-immunoprecipitation, and immunofluorescence assays were used to elucidate the mechanism. Results Overexpression of CDK1 in ACC cell lines promoted proliferation and induced the epithelial-to-mesenchymal transition (EMT), whereas knockdown of CDK1 expression inhibited growth of ACC cell lines. The CDK1 inhibitor, cucurbitacin E (CurE), had the best inhibitory effect with good time-and dose-dependent activity both in vitro and in vivo. CurE had a greater inhibitory effect on ACC xenografts in nude mice than mitotane, without obvious adverse effects. Most importantly, combined treatment with CurE and mitotane almost totally eliminated ACC tumors. With respect to mechanism, CDK1 facilitated the EMT of ACC cells via Slug and Twist and locked ACC cells into the G2/M checkpoint through interaction with UBE2C and AURKA/B. CDK1 also regulated pyroptosis, apoptosis, and necroptosis (PANoptosis) of ACC cells through binding with the PANoptosome in a ZBP1-dependent way. Conclusions CDK1 could be exploited as an essential therapeutic target of ACC via regulating the EMT, the G2/M checkpoint, and PANoptosis. Thus, CurE may be a potential candidate drug for ACC therapy with good safety and efficacy, which will meet the great need of patients with ACC.

Pulp regeneration with stem cells is a promising alternative for treating periapical and pulp diseases of young permanent teeth. The aim of this study was to characterize decellularized dental pulp extracellular matrix (dECM) and investigate whether bone morphogenetic protein 4 (BMP4) regulates dental pulp stromal cells (DPSC)-mediated pulp regeneration combined with dECM. Dental pulp isolated from healthy third molars was decellularized with 10% sodium dodecyl sulfate (SDS) and Triton X-100. H&E staining, DAPI staining and electron microscopy were used to observe the dECM structure. The Cell Counting Kit-8 assay was used to analyse cell proliferation. Recombinant adenovirus was used to overexpress BMP4 in DPSCs. The cells were cultured in dECM and dECM + three-dimensional (3D) Vitrogel systems, and bone/dentin/angiogenesis marker expression was evaluated by real-time polymerase chain reaction (RT-PCR) and ALP staining. DPSCs mixed with dECM and BMP4 were transplanted into nude mice, and pulp-like tissue formation was evaluated. The expression of osteogenic and angioblastic genes was increased, and pulp-like tissue formed in vivo. Thus, dECM promotes DPSC proliferation, BMP4 and dECM together accelerate pulp-like tissue formation by DPSCs in vitro.

Penoxsulam (PNX) and silver nanoparticles (AgNPs) are likely to coexist in soils due to continuous use. However, the ecotoxicity of PNX in earthworms and the effect of AgNPs on PNX toxicity are unknown. Therefore, the toxicity of PNX (0.05, 0.5, and 2.5 mg/kg) single and co-exposure with AgNPs (10 mg/kg) after 28 and 56 days on Eisenia fetida (E. fetida) was investigated from biochemical, genetic, histopathological, and transcriptomic aspects. The results showed that the low concentration of PNX (0.05PNX) had almost no effect on the biochemical level of E. fetida. However, the addition of AgNPs resulted in 0.05PNX causing E. fetida to produce excessive reactive oxygen species, and the activity of antioxidant and detoxification enzymes were interfered, resulting in lipid peroxidation and DNA damage. From the genetic level, even the lowest concentration of PNX can significantly interfere with the expression of functional genes, thus inducing oxidative stress and apoptosis and inhibited reproductive behavior in E. fetida. The integrated biomarker response results at the biochemical and genetic levels showed that the comprehensive toxicity of PNX and PNX + AgNPs on E. fetida was PNX dose-dependent. And the toxicity of all co-exposure groups was greater than that of the PNX only exposure groups. Furthermore, the addition of AgNPs significantly increased the damage of PNX on E. fetida intestinal tissue. Meanwhile, transcriptomic analysis showed that PNX + AgNPs had a greater effect on E. fetida than PNX single, and multiple pathways related to oxidative stress, inflammation, and cellular process regulation were disturbed. These results provide a basis for comprehensive evaluation of the ecotoxicity of PNX and confirm that the AgNPs does increase the ecotoxicity of PNX in soil environment.

Major yolk proteins (MYPs), one class of the main abundant proteins in sea cucumber body wall, seem to garner more attention in recent years. Herein, a method using multiple reactions monitoring mass spectrometry (MRM-MS) was deliberatively developed to perform quantification analysis of three MYPs, i.e. BAH79576.1, BAH79577.1 and PIK45784.1. Contents of MYPs in body wall of fresh and dried sea cucumbers as well as in waste liquid of boiling and steaming were determined using their corresponding signature peptides of VDEFTGIVGSLR, KLDMYPPPLAR, LDMYPPPLAR, and SGHGEVMFVDSK. The loss of MYPs in the processing of sea cucumbers was directly verified by quantitation data of MYPs in sea cucumber body wall and the waste liquids. This study not only evidenced the loss of MYPs during the processing of sea cucumbers, but also implicated the potential of recycling MYPs from the processing waste water, providing helpful suggestions in maximizing the value of sea cucumbers.

Janus Kinases (JAKs) have emerged as an important drug target for the treatment of a number of immune disorders due to the central role that they play in cytokine signalling. 4 isoforms of JAKs exist in mammalian cells and the ideal isoform profile of a JAK inhibitor has been the subject of much debate. JAK3 has been proposed as an ideal target due to its expression being largely restricted to the immune system and its requirement for signalling by cytokine receptors using the common γ-chain. Unlike other JAKs, JAK3 possesses a cysteine in its ATP binding pocket and this has allowed the design of isoform selective covalent JAK3 inhibitors targeting this residue. We report here that mutating this cysteine to serine does not prevent JAK3 catalytic activity but does greatly increase the IC50 for covalent JAK3 inhibitors. Mice with a Cys905Ser knockin mutation in the endogenous JAK3 gene are viable and show no apparent welfare issues. Cells from these mice show normal STAT phosphorylation in response to JAK3 dependent cytokines but are resistant to the effects of covalent JAK3 inhibitors. These mice therefore provide a chemical-genetic model to study JAK3 function.

As a class of synthetic sulfur drugs, sulfonamides (SAs) have been used to treat diseases and promote organism growth. Different concentrations of SAs have been detected in the water environment, which has threatened the ecological environment. In this study, the contamination of 9 SAs in water, sediments, and 8 fish species from the Hangbu-Fengle River, China, were analyzed using UPLC-MS/MS. The total SA concentrations in surface water, sediments, and fish were ND-5.064 ng/L, ND-5.052 ng/g dry weight (d.w.), and ND-1.42 ng/g wet weight (w.w.), respectively. The major compounds were sulfadiazine (SDZ), sulfamerazine (SMZ), and sulfamethoxazole (SMX) in water and fish. The SA levels of in fish from different habitat preferences revealed a spatial difference, with the order of demersal species > pelagic species. Moreover, the SA concentrations were affected by trophic guilds, indicating their decrease in the order of piscivorous fish > omnivorous fish > planktivorous fish > herbivorous fish. The obtained bioaccumulation factors showed that SMZ and SMX have strong bioenrichments in Ophiocephalus argus Cantor and Pelteobagrus fulvidraco. The risk assessment indicated that SAs did not pose significant health threats to the organisms. This research is the first report of SA contamination in the Hangbu-Fenle River, which can provide an important scientific basis for their pollution prevention and ecological risk assessment in the aquatic environment.

The goal of this work was to develop a sensitive and accurate method based on high performance liquid chromatography with tandem mass spectrometry (LC-MS) detection for determining the concentration of the Sinomenine derivative SWX in plasma and tissue of rat. Chromatographic separation was achieved on an Agilent SB-C18 (2.1*50 mm, 2.7 µm) column. The mobile phase consisted of acetonitrile (solvent A) and 0.2 % formic acid in water (solvent B) with gradient elution as follows: 0-3 min, 50-90 % A, 3 min-3.01 min, 90-50 % A, 90 % A in 3.01 min-10 min. The flow rate was set to 0.3 mL/min. The column temperature was 40 °C, and the sample injection volume was 10 µL. The calibration curve had good linearity in the range of 10 ng/mL to 600 ng/mL. Biological samples were prepared by protein precipitation with acetonitrile. The area under the curve (AUC) and the peak drug concentration (C

Polyhalogenated carbazoles (PHCZs) are widely present in the environment, and their health risks are of increasing concern. Available studies primarily confirm their dioxin-like toxicity mechanism based on biomarkers, such as aryl hydrocarbon receptor (AHR) and CYP1A1, while few studies have investigated their actual toxic effects at the level of individual organisms. In the present study, the developmental toxicity of two typical PHCZs with a high detection rate and high concentration in the environment (3,6-dichlorocarbazol (3,6-DCCZ) and 3,6-dibromocarbazole (3,6-DBCZ)) was investigated based on a fish embryo acute toxicity test (FET, zebrafish) and transcriptomics analysis. The 96 h LC

Type 2 diabetes mellitus (T2DM) markedly impairs human health. During T2DM development, some patients experience cognitive dysfunction and behavioral deficits, which are characterized by neuronal injury and memory loss. It has been reported that the incidence of dementia in middle-aged and elderly patients with diabetes is significantly higher than that in normal elderly patients. Currently, the pathogenesis of cognitive dysfunction in diabetes remains unknown, and there is no standard or specific method to diagnose the disease in clinical practice. Evidence has shown that fish oil (FO) can alleviate depressive-like behaviors by attenuating neuroinflammation in a rat model, and improve cognitive dysfunction by inhibiting apoptosis. Therefore, it is reasonable to speculate that FO may reduce cognitive impairment by attenuating neuroinflammation in diabetic rats. In the present study, we investigated the effects of FO supplementation on cognitive dysfunction in a streptozotocin-induced diabetic rat model. FO administration for 10 weeks improved spatial learning and memory as evaluated by performance in the Morris water maze (MWM). Besides, FO significantly improved the morphology of neurons in the hippocampus and cortex of diabetic rats and reduced the neuronal nuclear condensation. Moreover, FO decreased the mRNA expression of IL-1β, IL -6, and TNF-α and increased the mRNA expression of IL-4 and IL-10 in the cortex and hippocampus. FO also attenuated the brain inflammatory cascade and simultaneously reduced diabetes-induced oxidative stress. In addition, FO increased the protein expression of Nrf2 and HO-1 in cortex and hippocampus of diabetic rats. These results provide a novel horizon for the study of neuroprotective effect of FO and further clarify the connections among inflammation, oxidative stress and diabetes-induced cognitive impairment.

Doublecortin like kinase 1 (DCLK1) is an understudied kinase that is upregulated in a wide range of cancers, including pancreatic ductal adenocarcinoma (PDAC). However, little is known about its potential as a therapeutic target. We used chemoproteomic profiling and structure-based design to develop a selective, in vivo-compatible chemical probe of the DCLK1 kinase domain, DCLK1-IN-1. We demonstrate activity of DCLK1-IN-1 against clinically relevant patient-derived PDAC organoid models and use a combination of RNA-sequencing, proteomics and phosphoproteomics analysis to reveal that DCLK1 inhibition modulates proteins and pathways associated with cell motility in this context. DCLK1-IN-1 will serve as a versatile tool to investigate DCLK1 biology and establish its role in cancer. A highly selective inhibitor of the DCLK1/2 kinases is used to uncover the consequences of DCLK1 inhibition on viability, phosphosignaling and the transcriptome in patient-derived organoid models of pancreatic ductal adenocarcinoma.

Recently, several targeted agents have been developed for specific subsets of patients with acute myeloid leukaemia (AML), including midostaurin, the first FDA‐approved FLT3 inhibitor for newly diagnosed patients with FLT3 mutations. However, in the initial Phase I/II clinical trials, some patients without FLT3 mutations had transient responses to midostaurin, suggesting that this multi‐targeted kinase inhibitor might benefit AML patients more broadly. Here, we demonstrate submicromolar efficacy of midostaurin in vitro and efficacy in vivo against wild‐type (wt) FLT3‐expressing AML cell lines and primary cells, and we compare its effectiveness with that of other FLT3 inhibitors currently in clinical trials. Midostaurin was found to synergize with standard chemotherapeutic drugs and some targeted agents against AML cells without mutations in FLT3. The mechanism may involve, in part, the unique kinase profile of midostaurin that includes proteins implicated in AML transformation, such as SYK or KIT, or inhibition of ERK pathway or proviability signalling. Our findings support further investigation of midostaurin as a chemosensitizing agent in AML patients without FLT3 mutations.

Mutations in the E3 ubiquitin ligase CBL, found in several myeloid neoplasms, lead to decreased ubiquitin ligase activity. In murine systems, these mutations are associated with cytokine‐independent proliferation, thought to result from the activation of hematopoietic growth receptors, including FLT3 and KIT. Using cell lines and primary patient cells, we compared the activity of a panel of FLT3 inhibitors currently being used or tested in AML patients and also evaluated the effects of inhibition of the non‐receptor tyrosine kinase, SYK. We show that FLT3 inhibitors ranging from promiscuous to highly targeted are potent inhibitors of growth of leukaemia cells expressing mutant CBL in vitro, and we demonstrate in vivo efficacy of midostaurin using mouse models of mutant CBL. Potentiation of effects of targeted FLT3 inhibition by SYK inhibition has been demonstrated in models of mutant FLT3‐positive AML and AML characterized by hyperactivated SYK. Here, we show that targeted SYK inhibition similarly enhances the effects of midostaurin and other FLT3 inhibitors against mutant CBL‐positive leukaemia. Taken together, our results support the notion that mutant CBL‐expressing myeloid leukaemias are highly sensitive to available FLT3 inhibitors and that this effect can be significantly augmented by optimum inhibition of SYK kinase.

Lineage-defining transcription factors (TFs) are compelling targets for leukemia therapy, yet they are among the most challenging proteins to modulate directly with small molecules. We previously used CRISPR screening to identify a salt-inducible kinase 3 (SIK3) requirement for the growth of acute myeloid leukemia (AML) cell lines that overexpress the lineage TF myocyte enhancer factor (MEF2C). In this context, SIK3 maintains MEF2C function by directly phosphorylating histone deacetylase 4 (HDAC4), a repressive cofactor of MEF2C. In this study, we evaluated whether inhibition of SIK3 with the tool compound YKL-05-099 can suppress MEF2C function and attenuate disease progression in animal models of AML. Genetic targeting of SIK3 or MEF2C selectively suppressed the growth of transformed hematopoietic cells under in vitro and in vivo conditions. Similar phenotypes were obtained when cells were exposed to YKL-05-099, which caused cell-cycle arrest and apoptosis in MEF2C-expressing AML cell lines. An epigenomic analysis revealed that YKL-05-099 rapidly suppressed MEF2C function by altering the phosphorylation state and nuclear localization of HDAC4. Using a gatekeeper allele of SIK3, we found that the antiproliferative effects of YKL-05-099 occurred through on-target inhibition of SIK3 kinase activity. Based on these findings, we treated 2 different mouse models of MLL-AF9 AML with YKL-05-099, which attenuated disease progression in vivo and extended animal survival at well-tolerated doses. These findings validate SIK3 as a therapeutic target in MEF2C-addicted AML and provide a rationale for developing druglike inhibitors of SIK3 for definitive preclinical investigation and for studies in human patients.

For the first time, both spatial and temporal effects of fish feed on changes in abundance of antibiotic resistance genes (ARGs) were investigated in South Korea via quantifying ARGs and analyzing physicochemical parameters in the influent (IN) and effluent before (BF) and 30 min after (AF) the fish feeding time of sixteen flow-through fish farms. The absolute abundance of ARGs in AF samples was 5 times higher than in BF and 12 times higher than in IN samples. Values of physicochemical parameters such as ammonia, total nitrogen, suspended solids and turbidity in the effluent significantly increased by 21.6, 4.2, 2.6 and 1.65 times, respectively, after fish feeding. Spatially, the fish farms on Jeju Island exhibited higher relative abundance (3.02 × 10

Atrazine has been widely used in the world and caused environmental pollution, especially soil pollution. When assessing the toxicity of atrazine in soil, most studies used standardized artificial soils, while few studies focused on the real soil environments. In the present study, three natural soils and artificial soil were selected as test soils to study and compare the toxicities of atrazine to Eisenia fetida. Acute toxicity of atrazine was determined by filter paper and soil tests. In chronic toxicity study, after atrazine exposure, the content of reactive oxygen species in Eisenia fetida significantly increased and showed a dose-response relationship. The activity changes of three antioxidant enzymes and glutathione transferase showed that atrazine had obvious oxidative stress effect on earthworms. The contents of malondialdehyde and 8-hydroxy deoxyguanosine in 0.1 and 1 mg/kg atrazine treatment groups were significantly higher than the control, indicating that medium and high concentrations of atrazine could cause lipid and DNA damage in Eisenia fetida. The acute toxicity results and the integrated biomarker response index for chronic toxicity indicated that the toxicity order of atrazine was: red clayfluvo-aquic soilartificial soilblack soil, and that the toxicity of atrazine in artificial soil was not representative of its toxicity in real soil environment. The results of correlation analysis showed that three soil property parameters of organic carbon, organic matter and sand were most related to the toxicity of atrazine.

Butyl benzyl phthalate (BBP), a typical phthalate plasticizer, is frequently detected in aquatic environments, but its possible effects on fish liver are unknown. In this study, adult zebrafish were exposed to 5-500 μg/L BBP and cultured for 28 days. The toxicity mechanism of environmentally relevant concentrations of BBP in the liver was explored using integrated biomarker response (IBR), molecular docking, and histopathological analysis, based on the tests of oxidative stress, apoptosis, and tissue damage, respectively. The results revealed that exposure to 500 μg/L BBP caused lipid peroxidation and DNA damage and induced inflammatory responses in the liver and intestinal tissues. The accumulation of reactive oxygen species (ROS) is the primary manifestation of BBP toxicity and is accompanied by changes in the activities of antioxidant and detoxification enzymes. Notably, the pro-apoptotic genes (p53 and caspase-3) were still significantly upregulated in the 50 μg/L and 500 μg/L treatment groups on day 28. Moreover, BBP interfered with apoptosis by forming a stable complex with apoptosis proteins (P53 and Caspase-3). Our findings are helpful for understanding the toxicity mechanisms of BBP, which could further promote the assessment of the potential environmental risks of BBP.

Ionic liquids (ILs) are composed of only anions and cations and are liquid solvents at room temperature. Different functional groups were introduced into the ILs, conferring them with specific functions or purposes and thus forming special ILs, namely task-specific ILs (TSILs). Imidazolium-based ILs are the most widely used ILs in industrial production. To date, there have been some studies on the toxic effects of ILs on different organisms. However, the effect of functionalized groups on the toxicity of ILs is still unclear. In the present study, zebrafish were used as model organisms to study the toxic effects of 1-ethyl-3-methylimidazolium nitrate ([C

To explore the contribution of secondary bonds on storage stability of RSC, urea, n-propanol and sodium nitrite (NaNO

Cadmium (Cd) is one of the most important marine environmental pollutants that can cause oxidative damage and apoptosis in living organisms, and mitochondria are the key target cell organelles of Cd toxicity. In this study, we investigated the effect of Cd on the mitochondria of Meretrix meretrix gill cells and the underlying mechanism of mitochondria-mediated apoptosis following exposure to the metal. Exposure of the clams to 1.5, 3, 6 and 12 mg L−1 Cd2+ solutions led to swollen mitochondria compared with the untreated clams. The mitochondria also became vacuolated at the higher Cd2+ concentrations. Biochemical assays showed that monoamine oxidase (MAO) activity and mitochondrial membrane potential (Δψm) increased at 1.5 mg L−1 Cd2+, but decreased at higher Cd2+ concentrations, while the activities of malate dehydrogenase (MDH) and cytochrome oxidase (CCO) and the scavenging capacities of anti-superoxide anion (ASA) and anti-hydroxy radical (AHR) all decreased with increasing Cd2+ concentrations. Significant increases in the levels of MDA and H2O2 as well as in the activity levels of caspase-3, -8, and -9 were also observed in the Cd2+-treated clams. The results implied that Cd might induce apoptosis in M. meretrix via the mitochondrial caspase-dependent pathway.

Bone formation and resorption are typically coupled, such that the efficacy of anabolic osteoporosis treatments may be limited by bone destruction. The multi-kinase inhibitor YKL-05-099 potently inhibits salt inducible kinases (SIKs) and may represent a promising new class of bone anabolic agents. Here we report that YKL-05-099 increases bone formation in hypogonadal female mice without increasing bone resorption. Postnatal mice with inducible, global deletion of SIK2 and SIK3 show increased bone mass, increased bone formation, and, distinct from the effects of YKL-05-099, increased bone resorption. No cell-intrinsic role of SIKs in osteoclasts was noted. In addition to blocking SIKs, YKL-05-099 also binds and inhibits CSF1R, the receptor for the osteoclastogenic cytokine M-CSF. Modeling reveals that YKL-05-099 binds to SIK2 and CSF1R in a similar manner. Dual targeting of SIK2/3 and CSF1R induces bone formation without concomitantly increasing bone resorption and thereby may overcome limitations of most current anabolic osteoporosis therapies.

Xiaoxuming decoction (XXMD) is a traditional Chinese herbal medicine (CHM) that is used for the treatment of stroke in China. Stroke injury damages the cerebral vasculature and disrupts the autoregulation of vasoconstriction and vasodilatation, which is crucial for maintaining constant cerebral blood flow (CBF). It has been reported that XXMD exerts a positive effect on cerebral circulation in animal models of stroke. However, the mechanisms underlying the regulatory effect of XXMD on vascular tone, and the interactions among the multiple components of XXMD, remain unclear. In this study, XXMD was found to induce relaxation of the basilar artery rings of rats precontracted by 5-hydroxytryptamine (5-HT) in vitro, in a dose-dependent manner. The modulation of vascular tone and the process of cerebral ischemia are mediated via the interactions between G protein-coupled receptors (GPCRs) and their ligands, including 5-HT, angiotensin II (Ang II), and urotensin II (UII). Thus, the potential synergistic effects of the different components of XXMD on the regulation of vasoconstriction and vasodilation were further investigated by molecular docking based on network pharmacology. We constructed and analyzed a database comprising 963 compounds of XXMD and studied the interactions between five vascular GPCRs (5-HT1A receptor (5-HT1AR), 5-HT1B receptor (5-HT1BR), Ang II type 1 receptor (AT1R), beta 2-adrenergic receptor (β2-AR), and UII receptor (UTR)) and the various herbal constituents of XXMD using molecular docking. By constructing and analyzing the compound-target networks of XXMD, we found that Glycyrrhizae Radix et Rhizoma, Ginseng Radix et Rhizoma, and Paeoniae Radix Alba were the three major herbs that contained a large number of compounds with high docking scores. We additionally observed that several constituents of XXMD, including gallotannin, liquiritin apioside, nariutin, 1,2,3,4,6-pentagalloylglucose, folic acid, and ginsenoside Rb1, targeted multiple vascular GPCRs. Moreover, the interactions between the components of XXMD and the targets related to vascular tone constituted the comprehensive cerebrovascular regulatory function of XXMD and provided a material basis of the vasoregulatory function of XXMD. The study reports the contributions of various components of XXMD to the regulatory effects on vascular tone and provides scientific evidence for the multicomponent and multitargeting characteristics of XXMD.

The role of apolipoproteins in cardiovascular disease has been well investigated, but their participation in cancer has only been explored in a few published studies which showed a close link with certain kinds of cancer. In this review, we focused on the function of different kinds of apolipoproteins in cancers, autophagy, oxidative stress, and drug resistance. The potential application of apolipoproteins as biomarkers for cancer diagnosis and prognosis was highlighted, together with an investigation of their potential as drug targets for cancer treatment. Many important roles of apolipoproteins and their mechanisms in cancers were reviewed in detail and future perspectives of apolipoprotein research were discussed., We focused on roles of different kind apolipoproteins in cancers, autophagy, oxidative stress, resistance of drugs and hypolipidemic agents and cancers. In addition, apolipoproteins as drug targets of cancer treatment and biomarkers for cancer diagnosis and prognosis were discussed. Future perspectives of apolipoproteins research were described in the end.

The purpose of the present study was to study the effects of resveratrol on cognitive function in rats with vascular dementia and to investigate the molecular mechanisms of its neuroprotective effects. Forty‑five SD rats were randomly divided into 3 groups: The control group (Con group, n=15), the model group (VD group, n=15) and the resveratrol‑treated VD group (Res group, n=15). The VD rats (the VD group and the Res group) were generated by bilateral common carotid artery occlusion. The rats in the Res group received daily resveratrol treatment intraperitoneally for 4 weeks. Cognitive function was tested using the Morris water maze test. The levels of SOD and MDA (oxidative stress indicators) were detected by ELISA kits. The protein expression of Bax, Bcl‑2 and caspase‑3 was detected by western blotting. Compared with the rats in the Con group, the rats in the VD group exhibited decreased cognitive function, significantly increased hippocampal content of MDA, Bax and caspase‑3 (P

In recent years, pharmaceuticals and personal care products (PPCPs) that remain in the environment have become increasingly important. Carbamazepine (CBZ) is a widely used antiepileptic drug that has a potential impact on the environment due to its Physico-chemical properties, which are rarely eliminated in conventional water treatment. Daphnia magna Straus (DMS) is a fundamental link of aquatic ecosystem chain. The influence of CBZ toxicity on DMS can effectively reflect the effects of CBZ toxicity on the aquatic environment. In this study, DMS was used as a subject to assess the chronic effects of CBZ exposure. It was found that after 21 days of CBZ exposure, the breeding frequency, the total number of eggs laid, body length, and intrinsic growth rate of DMS decreased with increasing CBZ concentrations. Maximum reductions of 69% in fecundity and 60% in fertility were observed at 0.5 mg/L CBZ, while a maximum reduction of 60% in body length was observed at 0.001 mg/L CBZ concentration. The integrated biomarker response version 2 (IBRv2) analysis suggests that with the increase in CBZ concentration, the overall negative effect of CBZ on DMS was enhanced.

Phthalates (phthalic acid esters) have been widely applied as plasticizers. They are ubiquitous contaminants in soils, thereby posing a threat to human health. In this study, ecotoxicological effects of three typical PAEs (dimethyl phthalate-DMP, di-n-octyl phthalate-DOP and butyl benzyl phthalate-BBP) were investigated. As a biological indicator, earthworms (Eisenia fetida) were exposed to phthalates at various doses (0, 0.1, 1, 10 and 50 mg/kg) for different times (7, 14, 21, and 28 d). We evaluated the effects of phthalates on reactive oxygen species (ROS) generation, antioxidant enzymes (superoxide dismutase-SOD, peroxidase-POD and catalase-CAT) activities, glutathione S-transferase enzyme (GST) activity, malondialdehyde (MDA) content and DNA damage. Results showed that ROS content increased with increasing phthalates, whereas ROS content generally increased and then decreased with exposure time. However, antioxidant enzymes activities in earthworms displayed different trends. The GST activity in high-dose treatment group was significantly activated. For DMP and DOP, lipid peroxidation mainly occurred between 14 and 28 d, while for BBP, it primarily existed after 7 d and then disappeared after 28 d. Besides, comet assay indicated that there was a dose-response relationship between the DNA damage and phthalate dose, following DMP > DOP > BBP. Given their toxicity, it is important to understand the mechanisms associated with their eco-toxicity and to reduce their adverse impacts on the environment. Keywords: Phthalic acid esters, ROS, Antioxidant enzymes, Lipid peroxidation, DNA damage

Few studies have focused on the association between previous particulate matter (PM) exposure and antioxidant defense response to a haze challenge. In this study, a combined exposure model was used to investigate whether and how PM exposure history affected the antioxidant defense response to haze episodes. At first, C57BL/6 male mice were randomly assigned to three groups and exposed for 5 weeks to whole ambient air, ambient air containing a low (≤75 μg/m3) PM concentration, and filtered air, which simulated different exposure history of high, relatively low, and almost zero PM concentrations. Thereafter, all mice underwent a 3-day haze exposure followed by a 7-day exposure to filtered air. The indexes involved in the primary and secondary antioxidant defense response were determined after pre-exposure and haze exposure, as well as 1 day, 3 days, and 7 days after haze exposure. Our research demonstrated repeated exposure to a high PM concentration compromised the antioxidant defense response and was accompanied by an increased susceptibility to a haze challenge. Conversely, mice with a lower PM exposure developed an oxidative stress adaption that protected them against haze challenge more efficiently and in a more timely manner than was the case in mice without PM exposure history.

Bone tissue engineering requires a combination of cells, efficient biochemical and physicochemical factors, and biocompatible scaffolds. In this study, we evaluated the potential use of injectable Matrigel as a scaffold for the delivery of rat dental follicle stem/precursor cells (rDFSCs) transduced by bone morphogenetic protein (BMP) 9 to enhance osteogenic differentiation in vitro and promote ectopic bone formation in vivo. Recombinant adenovirus was used to overexpress BMP9 in rDFSCs. Alkaline phosphatase activity was measured using a histochemical staining assay and a chemiluminescence assay kit. Quantitative real-time polymerase chain reaction was used to determine mRNA expression levels of bone-related genes including distal-less homeobox 5 (DLX5), osteopontin (OPN), osterix (Osx), and runt-related transcription factor 2 (Runx2). Matrix mineralization was examined by Alizarin Red S staining. rDFSCs proliferation was analyzed using the Cell Counting Kit-8 assay. Subcutaneous implantation of rDFSCs-containing Matrigel scaffolds was used, and micro-computed tomography analysis, histological evaluation, and trichrome staining of implants extracted at 6 weeks were performed. We found that BMP9 enhanced alkaline phosphatase activity and mineralization in rDFSCs. The expression of bone-related genes (DLX5, OPN, Osx, and Runx2) was also increased as a result of BMP9 stimulation. Micro-computed tomography analysis and histological evaluation revealed that the bone masses retrieved from BMP9-overexpressing rDFSCs were significantly more pronounced in those with than in those without Matrigel. Our results suggest that BMP9 effectively promote osteogenic differentiation of rDFSCs, and Matrigel facilitate BMP9-induced osteogenesis of rDFSCs in vivo.

Sphingolipid metabolites, such as ceramide, sphingosine and sphingosine-1-phosphate (S1P), play many important roles in cellular activities. Ceramide and sphingosine inhibit cell proliferation and induce cell apoptosis while S1P has the opposite effect. Maintaining a metabolic balance of sphingolipids is essential for growth and development of cells. Sphingosine kinase (SPHK) is an important regulator for keeping this balance. It controls the level of S1P and plays important roles in proliferation, migration, and invasion of cancer cells and tumor angiogenesis. There are two isoenzymes of sphingosine kinase, SPHK1 and SPHK2. SPHK1 is ubiquitously expressed in most cancers where it promotes survival and proliferation, while SPHK2 is restricted to only certain tissues and its functions are not well characterized. SPHK1 is currently considered as a novel target for the treatment of cancers. Targeting SPHK1 would provide new strategies for cancer treatment and improve the prognosis of cancer patients. Here we review and summarize the current research findings on the SPHK1-S1P axis in cancer from many aspects including structure, expression, regulation, mechanism, and potential inhibitors.

Great efforts have been devoted to assessing the effects of straw managements on greenhouse gas (GHG) emissions, global warming potential (GWP), and net economic budget in rice monoculture (RM). However, few studies have evaluated the effects of straw managements on GHG emissions and net ecosystem economic budget (NEEB) in integrated rice-crayfish farming (RC). Here, a randomized block field experiment was performed to comprehensively evaluate the effects of aquatic breeding practices (feeding or no feeding of forage) and straw managements (rice straw returning or removal) on soil NH4+-N and NO-3-N contents, redox potential (Eh), CH4 and N2O emissions, GWP, and NEEB of fluvo-aquic paddy soil in a rice-crayfish co-culture system in Jianghan Plain of China. We also compared the differences in CH4 and N2O emissions, GWP, and NEEB between RM and RC. Straw returning significantly increased CH4 and N2O emissions by 34.9-46.1% and 6.2-23.1% respectively compared with straw removal. Feeding of forage decreased CH4 emissions by 13.9-18.7% but enhanced N2O emissions by 24.4-33.2% relative to no feeding. Compared with RM treatment, RC treatment decreased CH4 emissions by 18.1-19.6% but increased N2O emissions by 16.8-21.0%. Moreover, RC treatment decreased GWP by 16.8-22.0% while increased NEEB by 26.9-75.6% relative to RM treatment, suggesting that the RC model may be a promising option for mitigating GWP and increasing economic benefits of paddy fields. However, the RC model resulted in a lower grain yield compared with the RM model, indicating that more efforts are needed to simultaneously increase grain yield and NEEB and decrease GWP under RC model.

Lung cancer is by far the leading cause of cancer death worldwide, and 85% of patients are diagnosed with non-small cell lung cancer (NSCLC), which is still very difficult to treat. Skp2 functions as an oncogene that participates in processes of many cancers. Here, we report a novel Skp2 inhibitor AAA-237 that binds to Skp2 protein and inhibits the proliferation of the NSCLC cells. We further investigated the anti-NSCLC mechanism of AAA-237 and found that it arrested the cell cycle at the G0/G1 phase by targeting Skp2 to reduce the degradation of p21Cip1 and p27Kip1 or by transcriptionally activating FOXO1 to increase the mRNA expression of p21Cip1 and p27Kip1. More importantly, we found that treatment of a high concentration AAA-237 could induce apoptosis of NSCLC cells and treatment of a low AAA-237 concentration for a longer time could induce senescence of NSCLC cells. Similar results were found in nude mice xenografted with A549 cells. AAA-237 inhibited tumor growth by inducing apoptosis and senescence in a dose-dependent manner. Considering these results, we propose that AAA-237 could be a promising therapeutic drug for treating patients with NSCLC.

Extraction processes significantly alter the structural and functional properties of polysaccharides. In this study, we extracted polysaccharides from Chroogomphis rutilus fruiting bodies (designated as CRP) using four methods, including hot water, ultrasound, microwave and sequential ultrasound-microwave, and designated these polysaccharides as CRP-H, CRP-M, CRP-U and CRP-UM, respectively. All CRPs were heteropolysaccharides with semblable monosaccharide types of glucose, mannose and galactose, mainly constituted of α- d -glucopyranosyl-(1 → 4). The extraction processes significantly affected the molecular weights, monosaccharide proportions, glycosidic bond ratios, branching degrees, triple-helix conformation and surface morphology of the CRPs. Among them, CRP-UM showed the highest yield and most potent antioxidative capacity in vitro and in HL-7702 cells, but the weakest activation of immunostimulatory response in RAW264.7 cells. In contrast, CRP-H exhibited the lowest yield but strongest immunostimulatory activity. Overall, microwave extraction could be utilized as a general and practical CRP extraction approach, based on its relatively high yield and bioactivities.

BACKGROUND AND AIM Chemotherapy drugs do not work well in esophageal squamous cell carcinoma (ESCC), and none of the targeted drugs have been applied in clinic. This study aims to identify effective targeted drugs and related biomarkers for the treatment of ESCC. METHODS The effect of 40 Food and Drug Administration-approved small-molecule inhibitors was first tested in five ESCC cell lines. CCK8 assays and xenografts derived from ESCC cell lines were performed to evaluate the anti-ESCC effects of inhibitors or chemotherapeutic agents in vitro and in vivo, respectively. Immunohistochemistry was utilized to analyze the p-EGFR expression in tissues. Western blot combining with gray analysis was conducted to detect the expression of interest protein. Flow cytometry and immunofluorescence assay were used to analyze apoptosis, cell cycle, and mitotic changes after drug treatment. RESULTS Afatinib showed remarkable effects on inhibiting ESCC cells with higher expression of p-EGFR. Results from combinatorial screening in ESCC cells expressing lower phosphorylation level of EGFR showed that paclitaxel and afatinib presented a significant synergistic inhibitory effect (P

The effects of microplastics (MPs) on terrestrial organisms remain poorly understood, even though soil is an important MPs sink. In this study, the earthworms Eisenia fetida were exposed to 0.25% (w/w) of industrial-grade high-density polyethylene (HDPE, 28-145, 133-415 and 400-1464 μm) and polypropylene (PP, 8-125, 71-383 and 761-1660 μm) MPs in an agricultural soil for 28 d. The results showed that HDPE and PP MPs with different size ranges can be ingested by E. fetida. Exposure to different size ranges of HDPE and PP MPs altered the activities of superoxide dismutase, catalase and glutathione S-transferase and induced an increase in the 8-hydroxy-2'-deoxyguanosine level in E. fetida, suggesting that MPs-induced oxidative stress occurred in E. fetida. A size and type-dependent toxicity of MPs to E. fetida was demonstrated by the integrated biological response index. In addition, to obtain detailed molecular information on the responses of E. fetida to MPs exposure, transcriptomic analysis was conducted for E. fetida from HDPE (28-145 μm) and PP (8-125 μm) treatment groups. Transcriptomic analysis identified 34,937 and 28,494 differentially expressed genes in the HDPE and PP MPs treatments compared with the control, respectively. And, exposure to HDPE and PP MPs significantly disturbed several pathways closely related to neurodegeneration, oxidative stress and inflammatory responses in E. fetida. This study provides important information for the ecological risk assessment of different size ranges and types of industrial-grade MPs.

Trifloxystrobin is a new type of fungicide, which is extensively used due to its excellent antifungal activity. In this study, oxidative stress and DNA damage induced by trifloxystrobin exposure was evaluated using Eisenia fetida at subchronic toxicity concentrations in artificial soil and brown soil (0.1–2.5 mg/kg). Throughout the exposure period (days 7, 28 and 56), six biochemical indicators including reactive oxygen species (ROS), antioxidant enzymes (SOD and CAT), glutathione S-transferase (GST), lipid peroxidation and DNA damage (8-hydroxydeoxyguanosine) were measured. In addition, the integrated biomarker response (IBR) index was calculated to make comparison of toxicological response between artificial and brown soils. Results indicated that trifloxystrobin can induce oxidative stress and DNA damage to earthworms with subchronic toxicity greater in brown soil compared to artificial soil as determined through integrated calculations for six biochemical indicators. Trifloxystrobin toxicological experiments in artificial soil may not accurately evaluate its toxicity in natural soil ecosystems, as the toxicity of trifloxystrobin to Eisenia fetida was underestimated. Refereed/Peer-reviewed

Lung cancer is the fastest growth rate of morbidity and mortality in nearly a decade, and remains difficult to treat. Furthermore, the molecular mechanisms underlying its development are still unclear. In this study, bioinformatics analysis showed that MELK was highly expressed in lung cancer and negatively correlated to the survival of lung adenocarcinoma (LUAD). Immunohistochemistry analysis of LUAD patient tissues revealed there were a high level of MELK expression in LUAD. Knockdown of MELK expression inhibits the migration and invasion of LUAD cells, which may be mediated by Twist1, Slug, MMP7, and N-catenin. Overexpression of MELK promoted the growth of LUAD cells in medium, 3D Matrigel, and nude mice. Inhibition of MELK by OTSSP167 arrested cycle of LUAD cells at G2/M phase via PLK1-CDC25C-CDK1 pathway, and triggered apoptosis-mediated pyroptosis. Together, these data indicate that MELK is critical for metastasis, mitotic progression, and programmed death of LUAD and may be a promising therapeutic target for LUAD.

Background DPP8 and DPP9 have been demonstrated to play important roles in multiple diseases. Evidence for increased gene expression of DPP8 and DPP9 in tubulointerstitium was found to be associated with the decline of kidney function in chronic kidney disease (CKD) patients, which was observed in the Nephroseq human database. To examine the role of DPP8 and DPP9 in the tubulointerstitial injury, we determined the efficacy of DPP8 and DPP9 on epithelial-to-mesenchymal transition (EMT) and tubulointerstitial fibrosis (TIF) as well as the underlying mechanisms. Methods We conducted the immunofluorescence of DPP8 and DPP9 in kidney biopsy specimens of CKD patients, established unilateral ureteral obstruction (UUO) animal model, treated with TC-E5007 (a specific inhibitor of both DPP8 and DPP9) or Saxagliptin (positive control) or saline, and HK-2 cells model. Results We observed the significantly increased expression of DPP8 and DPP9 in the renal proximal tubule epithelial cells of CKD patients compared to the healthy control subjects. DPP8/DPP9 inhibitor TC-E5007 could significantly attenuate the EMT and extracellular matrix (ECM) synthesis in UUO mice, all these effects were mediated via interfering with the TGF-β1/Smad signaling. TC-E5007 treatment also presented reduced renal inflammation and improved renal function in the UUO mice compared to the placebo-treated UUO group. Furthermore, the siRNA for DPP8 and DPP9, and TC-E5007 treatment decreased EMT- and ECM-related proteins in TGF-β1-treated HK-2 cells respectively, which could be reversed significantly by transduction with lentivirus-DPP8 and lentivirus-DPP9. Conclusion These data obtained provide evidence that the DPP8 and DPP9 could be potential therapeutic targets against TIF.

The non-enzymatic degradation of ready-to-eat sea cucumber (RSC) was closely related to the quality of sea cucumber products. When stored at 37 °C for 0-30 d, the hardness of RSC decreased by 86.7% and the proportion of free water increased by 12.71%. The content of free hydroxyproline increased from 8.33 μg/g to 24.12 μg/g. Label-free quantitative proteomics analysis showed that protein was prone to break at the sites of G, Q, N, D, and L, and the peptide bonds in QI, DL, NL, RI, EF and SY were much more liable to break. Edman degradation method showed that the breakage sites of RSC were at S, D, H, E, and V. NL, NA and NG calculated by B3LYP/6-31G(d) showed that the relative free energies in the initial cyclization step were 53.20, 143.53 and 78.10 kcal/mol, respectively, which may be the rate-determining step for peptide bond cleavage.

Azoxystrobin, a widely used broad-spectrum strobilurin fungicide, may pose a potential threat in agricultural ecosystems. To assess the ecological risk of azoxystrobin in real soil environments, we performed a study on the toxic effects of azoxystrobin on earthworms (Eisenia fetida) in three different natural soils (fluvo-aquic soil, black soil and red clay soil) and an artificial soil. Acute toxicity of azoxystrobin was determined by filter paper test and soil test. Accordingly, exposure concentrations of chronic toxicity were set at 0, 0.1, 1.0 and 2.5 mg kg−1. For chronic toxicity test, reactive oxygen species, activity of antioxidant enzymes (superoxide dismutase, catalase and peroxidase), detoxifying enzyme (glutathione transferase), level of lipid peroxidation (malondialdehyde) and level of oxygen damage of DNA (8-hydroxydeoxyguanosine) in earthworms were determined on the 7th, 14th, 21st, 28th, 42nd and 56th days after treatment. Both acute and chronic toxic results showed azoxystrobin exhibit higher toxicity in natural soil than in artificial soil, indicating that traditional artificial soil testing method underestimate ecotoxicity of azoxystrobin in a real agricultural environment on the earthworm population. Combining with the analysis of soil physicochemical properties, the present experiment provided scientific guidance for rational application of azoxystrobin in agricultural production systems.

Our study attempted to compare the efficacies of bone morphogenetic protein (BMP) 2, 6, and 9 in inducing osteogenic differentiation of preodontoblasts (PDBs). We immortalized PDBs by introducing a reversible SV40 T antigen-based immortalization system. Cell proliferation capability was examined by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. The effects of BMP2, 6, and 9 on the osteogenic differentiation of immortalized preodontoblasts (iPDBs) were measured by alkaline phosphatase (ALP) activity assays and alizarin red S staining. The expression of osteogenic markers was evaluated by semiquantitative real-time polymerase chain reaction analysis. To assess ectopic bone formation, rat-derived iPDBs were transfected in culture with adenoviral vectors designated Ad-BMP2, 6, and 9 and subcutaneously or intramuscularly injected into mice. Several BMPs retained endogenous expression in PDBs and regulated the mRNA expression of mineralized tissue-associated proteins. ALP activity and mineralized nodule formation were significantly increased in the Ad-BMP9-transfected group relative to the control group. In addition, the most significant hard tissue formation was in this group. The results indicated that BMP signaling was involved in the osteogenic differentiation of iPDBs. BMP9 could be an efficacious accelerant of the osteogenic differentiation of iPDBs.

In this study, the artificial soil poisoning method was used to explore the antioxidative stress mechanism and gene changes of earthworms (Eisenia foetida) after application of nitenpyram. The toxic effects of nitenpyram on earthworms were combined with the method called the second-generation integrated biomarker response index method (IBRv2) to be comprehensively analyzed by studying the reactive oxygen species (ROS) content, superoxide dismutase (SOD) activity, catalase (CAT) activity, glutathione S-transferase (GST) activity, malondialdehyde (MDA) content and DNA damage degree in earthworms. The results showed that the ROS content in the high-concentration (2.5 mg/kg) nitenpyram treatment group changed significantly. The changes of antioxidant enzymes in earthworms were also obvious. In terms of SOD enzyme activity, under the induction of nitenpyram, SOD activity in the 1 mg/kg and 2.5 mg/kg treatment groups was significantly enhanced. The concentration-treated group could all affect the activity of earthworm detoxifying enzyme GST. Earthworm DNA olive tail in the nitenpyram treatment group with different concentrations was mainly concentrated at low and medium levels at 21d, and the proportion was the largest during the whole exposure period, showing a significant dose-effect relationship. This study confirms that nitenpyram not only has a toxic effect on the physiological and biochemical indicators of earthworms, but also cannot be underestimated on its genetic level.

Present-day diagnostic tools and technologies for canine diseases and other vector-borne parasitic diseases hardly meet the requirements of an efficient and rapid diagnostic tool, which can be suitable for use at the point-of-care in resource-limited settings. Loop-mediated isothermal amplification (LAMP) technique has been always a method of choice in the development and validation of quick, precise, and sensitive diagnostic assays for pathogen detection and to reorganize point-of-care (POC) molecular diagnostics. In this study, we have demonstrated an efficient detection system for parasitic vector-borne pathogens like Ehrlichia canis and Hepatozoon canis by linking the LAMP assay to a smartphone via a simple, inexpensive, and a portable “LAMP box,” All the components of the LAMP box were connected to each other wirelessly. This LAMP box was made up of an isothermal heating pad mounted below an aluminum base which served as a platform for the reaction tubes and LAMP assay. The entire setup could be connected to a smartphone via an inbuilt Wi-Fi that allowed the user to establish the connection to control the LAMP box. A 5 V USB power source was used as a power supply. The sensitivity of the LAMP assay was estimated to be up to 10−6 dilution limit using the amplified, purified, and quantified specific DNA templates. It can also serve as an efficient diagnostic platform for many other veterinary infectious or parasitic diseases of zoonotic origin majorly towards field-based diagnostics. Supplementary Information The online version contains supplementary material available at 10.1007/s00436-021-07077-z.

With the wide use of mulch film and pesticides, mulch film-derived microplastics are very likely to produce combined effects with pesticides in agricultural soil. However, little is known about their combined toxicity on terrestrial organisms. This study aimed to investigate the combined toxicity of unused or farmland residual transparent low-density polyethylene mulch film-derived microplastics (MPs and MPs-aged, respectively) (550–1000 μm) and atrazine (ATZ; 0.02 and 2.0 mg/kg) on the earthworm (Eisenia fetida). After single and combined exposure to ATZ and microplastics for 28 d, the results showed an accumulation of reactive oxygen species, a decrease in superoxide dismutase, catalase, and glutathione-S-transferase activities, an increase in the malondialdehyde and 8-hydroxydeoxyguanosine levels, and abnormal expression of annetocin, heat shock protein 70, translationally controlled tumor protein and calreticulin genes. Integrated biological response (IBR) values calculated at the biochemical level indicated that the combined exposure to ATZ and microplastics, particularly to high concentrations of ATZ, induced greater oxidative stress in E. fetida compared with that of exposure to ATZ or microplastics alone. In addition, the IBR values calculated at the gene level did not show regular changes after combined exposure to ATZ and microplastics compared with those of a single exposure. The oxidative stress and abnormal expression of genes in E. fetida induced by MPs-aged were higher than those induced by MPs; a similar trend was observed for oxidative stress induced by MPs/MPs-aged + ATZ2.0, whereas an opposite trend was observed for the abnormal expression of genes in E. fetida induced by MPs/MPs-aged + ATZ0.02/ATZ2.0. Our results suggest that mulch film-derived microplastics have the potential to enhance the toxicity of ATZ within the soil environment.

The outbreak of COVID-19, the pandemic disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spurred an intense search for treatments by the scientific community. In the absence of a vaccine, the goal is to target the viral life cycle and alleviate the lung-damaging symptoms of infection, which can be life-threatening. There are numerous protein kinases associated with these processes that can be inhibited by FDA-approved drugs, the repurposing of which presents an alluring option as they have been thoroughly vetted for safety and are more readily available for treatment of patients and testing in clinical trials. Here, we characterize more than 30 approved kinase inhibitors in terms of their antiviral potential, due to their measured potency against key kinases required for viral entry, metabolism, or reproduction. We also highlight inhibitors with potential to reverse pulmonary insufficiency because of their anti-inflammatory activity, cytokine suppression, or antifibrotic activity. Certain agents are projected to be dual-purpose drugs in terms of antiviral activity and alleviation of disease symptoms, however drug combination is also an option for inhibitors with optimal pharmacokinetic properties that allow safe and efficacious co-administration with other drugs, such as antiviral agents, IL-6 blocking agents, or other kinase inhibitors.

BACKGROUND: There is a need to develop novel therapies which could be beneficial to prostate cancer (CaP) patients including those who are predisposed to poor outcome such as African-Americans. This study investigates the role of ROBO1-pathway in predicting outcome and race-based disparity in CaP patients. METHODS AND RESULTS: Aided by RNA sequencing-based DECIPHER-testing and IHC analysis of tumors we show that ROBO1 is lost during the progressive stages of CaP, a prevalent feature in African-Americans. We show that the loss of ROBO1 predicts high-risk of recurrence, metastasis and poor outcome of ADT in RP-treated patients. These data identified an aggressive ROBO1(deficient)/DOCK1(+ve) sub-class of CaP. Combined genetic and IHC data showed that ROBO1 loss is accompanied by DOCK1/Rac1 elevation in Grade-III/IV primary-tumors and Mets. We observed that the hypermethylation of ROBO1-promoter contributes to loss of expression that is highly prevalent in African-Americans. Because of limitations in restoring ROBO1 function, we asked if targeting the DOCK1 could be an ideal strategy to inhibit progression or treat ROBO1(deficient) metastatic-CaP. We tested the pharmacological efficacy of CPYPP, a selective inhibitor of DOCK1 under in vitro and in vivo conditions. Using ROBO1(−ve) and ROBO1(+ve) CaP models, we determined the EC(50) for growth. DOCK1-inhibitor treatment significantly decreased the (i) Rac1-GTP/β-catenin activity, (ii) transmigration of ROBO1(deficient) cells across endothelial lining and (iii) metastatic spread of ROBO1(deficient) cells through the vasculature of transgenic(fl) Zebrafish model. CONCLUSION: We suggest that ROBO1 status forms as predictive biomarker of outcome in high-risk populations such as African-Americans and DOCK1-targeting therapy has a clinical potential for treating metastatic-CaP.

The programmed cell death 1 (PD-1) receptor on the surface of immune cells is an immune checkpoint molecule that mediates the immune escape of tumor cells. Consequently, antibodies targeting PD-1 have shown efficacy in enhancing the antitumor activity of T cells in some types of cancers. However, the potential effects of PD-1 on tumor cells remain largely unknown. Here, we show that PD-1 is expressed across a broad range of tumor cells. The silencing of PD-1 or its ligand, PD-1 ligand 1 (PD-L1), promotes cell proliferation and colony formation in vitro and tumor growth in vivo. Conversely, overexpression of PD-1 or PD-L1 inhibits tumor cell proliferation and colony formation. Moreover, blocking antibodies targeting PD-1 or PD-L1 promote tumor growth in cell cultures and xenografts. Mechanistically, the coordination of PD-1 and PD-L1 activates its major downstream signaling pathways including the AKT and ERK1/2 pathways, thus enhancing tumor cell growth. This study demonstrates that PD-1/PD-L1 is a potential tumor suppressor and potentially regulates the response to anti-PD-1/PD-L1 treatments, thus representing a potential biomarker for the optimal cancer immunotherapeutic treatment.

Vaccines and antivirals to combat dengue, Zika, and other flavivirus pathogens present a major, unmet medical need. Vaccine development has been severely challenged by the antigenic diversity of these viruses and the propensity of non-neutralizing, cross-reactive antibodies to facilitate cellular infection and increase disease severity. As an alternative, direct-acting antivirals targeting the flavivirus envelope protein, E, have the potential to act via an analogous mode of action without the risk of antibody-dependent enhancement of infection and disease. We previously discovered that structurally diverse small molecule inhibitors of the dengue virus E protein exhibit varying levels of antiviral activity against other flaviviruses in cell culture. Here, we demonstrate that the broad-spectrum activity of several cyanohydrazones against dengue, Zika, and Japanese encephalitis viruses is due to specific inhibition of E-mediated membrane fusion during viral entry and provide proof of concept for pharmacological inhibition of E as an antiviral strategy in vivo.

Fluoxastrobin is one of the most widely used strobilurin fungicides, however, application of the fungicides may result in soil residues leading to environmental damage including oxidative stress and damage to sentinel organisms (i.e. earthworms). While this has been demonstrated in artificial soil, the biochemical response of Eisenia fetida exposed to fluoxastrobin in natural soils is unclear. This study utilized three typical natural soils (fluvo-aquic soils, red clay, and black soils) to evaluate the biochemical response of Eisenia fetida exposed to fluoxastrobin (0.1, 1.0, 2.5 mg/kg) including the production of reactive oxygen species, impact on three enzyme activities, lipid peroxidation, and 8-hydroxydeoxyguanosine after a 4-week exposure. The effects of fluoxastrobin on Eisenia fetida in different soils were assessed using an integrated biomarker response (IBR). The findings may be possible to state that the toxic effects of fluoxastrobin in artificial cannot exactly represent that in natural soils. Specifically, the fluoxastrobin subchronic toxicity was highest in red clay and lowest in black soil among the three natural soils. Furthermore, the 8-OHdG content was more sensitive to fluoxastrobin in all six environmental indicators of the present study. Refereed/Peer-reviewed

Dibutyl phthalate (DBP) is one of the most widely used plasticizers with a high concentration in the water. Although the toxicity of DBP on aquatic organisms has become a significant concern in recent years, the effects of DBP on zebrafish (Danio rerio) brain is poorly understood. This study investigated the toxic effects of DBP exposure for 7, 14, 21 and 28 days on zebrafish brain. The results showed that DBP significantly stimulated SOD and CAT activities, increasing MDA and 8-OHdG contents. On the 28th day, the AChE inhibition rates in 0.08, 0.4, 2 mg·L-1 treatment were 13.4%, 11.9%, 14.7%. The trend of Cu/Zn-sod gene variation was consistent with SOD activity, showing “inhibition-activation-inhibition”. The expression of apoptotic genes (caspase-3, p53) showed “inhibition-activation-inhibition”. The integrated biomarker response (IBR) results showed that the IBR values were 4.37, 7.18 and 9.63 in 0.08, 0.4 and 2 mg·L-1 group on the 28th day, presenting a “dose-response” relationship. These findings confirmed that low concentration of DBP induced oxidative damage and genotoxicity in zebrafish brain, which provided an effective toxicological basis for phthalate pollution. Based on above studies, it is of great significance for assessing the harmful effects of DBP with low concentration on aquatic organisms.

Neurodegenerative diseases are closely related to neuroinflammation. Drugs targeting inflammation have been proved to be effective in many animal models. Salvianolic acid C (SalC) is a compound isolated from Salvia miltiorrhiza Bunge, a plant with reported effects of inhibiting inflammation. However, the anti-inflammation effects and biological mechanisms of SalC on LPS-stimulated neuroinflammation remain unknown. The aim of this paper was to study its protective effects and its anti-inflammation mechanisms. LPS was used both in vivo and in vitro to induce neuroinflammation in SD rats and microglia cells. MTT assay was carried out to detect cell viability. The levels of TNF‑α, IL‑1β, IL‑6, IL‑10 and PGE 2 were detected by ELISA method. The expressions of p‑AMPK, p‑NF‑κB p65, p‑IκBα, Nrf2, HO‑1 and NQO1 proteins were examined by Western blot analysis. The nuclear translocation of NF‑κB p65 was studied by immunofluorescence assay. The specific Nrf2 siRNA was used to clarify the interaction between Nrf2 and NF‑κB p65. The AMPK inhibitor Compound C was used study the upstream protein of Nrf2. Results showed that LPS induced the overexpression of inflammatory cytokines and mediated the phosphorylation and nuclear translocation of NF‑κB p65 in rat brains and microglia cells. SalC reversed the inflammatory response induced by LPS and inhibited the NF‑κB activation. SalC also upregulated the expression of p‑AMPK, Nrf2, HO‑1 and NQO1. But the anti-inflammation and NF‑κB inhibition effects of SalC were attenuated by transfection with specific Nrf2 siRNA or interference with the potent AMPK inhibitor Compound C. In conclusion, SalC inhibited LPS-induced inflammatory response and NF‑κB activation through the activation of AMPK/Nrf2 signaling both in vivo and in vitro.