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Nephrotoxicity is a major adverse reaction of cisplatin-based chemotherapy. Organic cation transporter 2 (OCT2) which is located on the basement membrane of human proximal renal tubules is responsible for the renal accumulation of cisplatin and its nephrotoxicity. This study aimed to investigate the protective effect of PPIs to CP-induced nephrotoxicity. Three kinds of PPIs including lansoprazole, omeprazole and rabeprazole (Rab) were co-administrated with CP to mice. In addition, OCT2-overexpressed HEK293, HK-2 and A549 cells were co-incubated with CP and PPIs. The results showed that PPIs can attenuate CP-induced increase of CRE, BUN and histological damage of kidney. Among the three PPIs, Rab was found with a superior protective effect. It significantly reduced the accumulation of CP in OCT2-overexpressed HEK293 cells and in the renal cortex tissues of mice, but not in HK-2 cells. Moreover, Rab reduced the expression levels of cleaved-caspase-3, RIPK1, RIPK3, MLKL and p-MLKL and the apoptosis rate of renal tubular cells induced by CP in vivo, but not in HK-2 cells. However, Rab increased the viability of CP-treated cells in a concentration-dependent manner and attenuated CP-induced apoptosis and necroptosis in OCT2 over-expressed HEK293 cells. Finally, we demonstrated that Rab have no influence on the antitumor effect of CP. In conclusion, Rab attenuate CP-induced nephrotoxicity mainly through inhibiting OCT2-mediated CP uptake, without interfering with its anti-tumor property of inducing apoptosis and necroptosis.

Umbelliferone (UMB), a natural coumarin compound, has been reported to possess anti-rheumatic effects on rheumatoid arthritis (RA) experimental models, but its potential role of UMB in regulating migration, invasion and inflammation of RA fibroblast-like synoviocytes (FLS) remain unclear. Herein, MTT assay was performed to confirm the non-cytotoxic concentrations (10, 20, and 40[Formula: see text][Formula: see text]M) and the treatment time (24[Formula: see text]h) of UMB on TNF-[Formula: see text]-stimulated RA FLS (MH7A cells) in vitro. Results of wound-healing, transwell and phalloidin staining assays revealed that UMB inhibited TNF-[Formula: see text]-induced migration, invasion and F-actin cytoskeletal reorganization in MH7A. Results of ELISA, western blot and gelatin zymography indicated that UMB decreased the productions of pro-inflammatory factors, including IL-1[Formula: see text], IL-6, IL-8, MMP-2 and MMP-9, and inhibited MMP-2 activity in TNF-[Formula: see text]-stimulated MH7A cells. In vivo, UMB (25[Formula: see text]mg/kg and 50[Formula: see text]mg/kg) relieved the joint damage and synovial inflammation in rats with adjuvant-induced arthritis (AIA). Mechanistically, UMB could suppress Wnt/[Formula: see text]-catenin signaling both in TNF-[Formula: see text]-induced MH7A cells and in AIA rat synovium, evidenced by decreasing Wnt1 protein level, activating GSK-3[Formula: see text] kinase by blocking GSK-3[Formula: see text] (Ser9) phosphorylation, and reducing the protein level and nuclear translocation of [Formula: see text]-catenin. Importantly, combined use of lithium chloride (a Wnt/[Formula: see text]-catenin signaling agonist) eliminated the inhibitory effects of UMB on migration, invasion and inflammation in vitro and the anti-arthritic effects of UMB in vivo. We concluded that UMB inhibited TNF-[Formula: see text]-induced migration, invasion and inflammation of RA FLS and attenuated the severity of rat AIA through its ability to block Wnt/[Formula: see text]-catenin signaling pathway.

Necroptosis is defined as a novel programmed cell necrosis that is mediated by receptor interacting serine-threonine protein kinase 1 (RIPK1) and other related signals. Necrosis, apoptosis and inflammation are commonly considered as the leading mechanism in acute kidney injury (AKI) induced by gentamicin (GEN), which is a useful antibiotic for treating the infection of Gram-negative bacterial. However, the necroptosis in the pathogenesis of GEN-induced AKI is unknown. In this study, to investigate the process and function of necroptosis in GEN-induced AKI, NRK-52E and HK-2 cells and SD rats were used as the models. The necroptosis-related proteins, including RIPK1, RIPK3, mixed lineage kinase domain-like (MLKL) and phosphorylated MLKL (p-MLKL), were all increasing time-dependently when GEN was continuously given. By using the RIPK1 inhibitor necrostatin-1 (NEC-1) and RIPK3 inhibitor (CPD42), the GEN-induced toxicity of tubular cells was alleviated. Moreover, it was validated that GEN-induced cell apoptosis and inflammation were attenuated after treating with NEC-1 or CPD42, both in vivo and in vitro. When MLKL was knocked down by siRNA, NEC-1 and CPD42 can not further protect the damage of tubular cells by GEN. Although the using of pan-caspase inhibitor Z-VAD significantly decreased GEN-induced apoptosis, it enhanced necroptosis and slightly promoted the decreased cell viability in GEN-treated cells, with the protective effects weaker than NEC-1 or CPD42. Finally, in vitro minimum inhibitory concentration (MIC) tests and bacteriostatic ring studies showed that NEC-1 did not interfere with the antibiotic effects of GEN. Thus, suppressing necroptosis can serve as a promising strategy for the prevention of GEN-induced nephrotoxicity.

Meng-yuan Zhou,1,* Li Cai,1,2,* Xiao-wen Feng,1 Yu-rong Mu,1 Bo Meng,1 Fang-yuan Liu,1 Rong Li1 1Inflammation and Immune Mediated Diseases Laboratory of Anhui Province, School of Pharmacy, Anhui Medical University, Hefei, 230032, Anhui Province, People’s Republic of China; 2Department of Pathology, School of Basic Medicine, Anhui Medical University, Hefei, 230032, Anhui Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Rong LiSchool of Pharmacy, Anhui Medical University, 81 Meishan Road, Hefei, 230032, Anhui Province, People’s Republic of ChinaTel +86 551-65172132Email aydlirong@163.comIntroduction: Previous studies have confirmed the pathologic role of synovial aquaporin 1 (AQP1) in rheumatoid arthritis (RA), but its associations with the abnormal biologic behaviors of fibroblast-like synoviocytes (FLS) remain unclear. Herein, we examined the roles of AQP1 in the proliferation, migration and invasion of TNF-α-stimulated RA FLS (MH7A cells) and explored the underlying mechanisms.Materials and Methods: Lentivirus-mediated AQP1 overexpression or silencing MH7A cells was constructed. Assays of MTT, flow cytometry (PI staining and Annexin V-PE/7-AAD staining), TMRM staining, wound-healing, transwell and phalloidin staining were performed to detect cell proliferation, cycle distribution, apoptosis, migration and invasion. The involvement of Wnt/β-catenin pathway was revealed by Western blot and β-catenin immunofluorescence staining.Results: AQP1 overexpression promoted cell proliferation of TNF-α-stimulated MH7A by facilitating transformation from G0/G1 to S phase and inhibiting cell apoptosis (ie, reduced apoptosis rates, raised mitochondrial membrane potential, increased Bcl-2 protein level and decreased levels of Bax and cleaved caspase 3 protein). Also, AQP1 overexpression increased the migration index as well as the numbers of migrated and invasive cells. Furthermore, AQP1 overexpression promoted the activation of Wnt/β-catenin pathway, and XAV939, an inhibitor of Wnt/β-catenin, canceled the above effects of AQP1 overexpression on MH7A cells. As expected, AQP1 silencing exhibited the opposite effects on TNF-α-stimulated MH7A cells, which could be reversed by LiCl, an activator of Wnt/β-catenin.Conclusion: AQP1 can affect the proliferation, migration and invasion of MH7A cells by Wnt/β-catenin signaling pathway, and AQP1 can be as a crucial determiner that can regulate RA FLS biologic behaviors.Keywords: aquaporin 1, rheumatoid arthritis, fibroblast-like synoviocytes, Wnt/β-catenin signaling pathway, biologic behaviors

BackgroundTertiary lymphoid structures (TLSs) have been proposed to assess the prognosis of patients with cancer. Here, we investigated the prognostic value and relevant mechanisms of TLSs in colorectal cancer liver metastases (CRCLM).Methods603 patients with CRCLM treated by surgical resection from three cancer centers were included. The TLSs were categorized according to their anatomic subregions and quantified, and a TLS scoring system was established for intratumor region (T score) and peritumor region (P score). Differences in relapse-free survival (RFS) and overall survival (OS) between groups were determined. Multiplex immunohistochemical staining (mIHC) was used to determine the cellular composition of TLSs in 40 CRCLM patients.ResultsT score positively correlated with superior prognosis, while P score negatively associated with poor survival (all p=0.029, p=0.047 and p=0.041, respectively), and the frequencies of Treg cells and M2 macrophages were positively correlated with P score, while the frequencies of Tfh cells were positively associated with T scores in intratumor TLSs (all pConclusionsThe distribution and abundance of TLSs is significantly associated with RFS and OS of CRCLM patients, and a novel immune class was proposed for predicting the prognosis of CRCLM patients.

In this study, the effect of corn oligopeptides (COPs) with liver protection activity on mice with hepatic fibrosis (HF) induced by carbon tetrachloride (CCl

Corn gluten meal (CGM) was hydrolyzed by Alcalase 2.4L and Protex 7L to obtain low-molecular-weight peptides. The antioxidant and angiotensin-I-converting enzyme (ACE) inhibitory activities of the peptides were determined by 2,2′-Azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) free radical scavenging activity, the JHH7 cell oxidative stress model, and the ACE inhibition assay, respectively. Corn peptides (CPs) had strong antioxidant capacity to scavenge ABTS radicals (1.44 ± 0.11 mmol/g), and significantly reduced reactive oxygen species and malondialdehyde levels in oxidized JHH7 cells, increasing superoxide dismutase, catalase, and glutathione peroxidase activities by 26.20%, 147.52%, and 43.20%, respectively, at the concentration of 800 μg/mL. CPs had high ACE inhibitory activity, with an IC50 value of 0.27 ± 0.05 mg/mL. Next, the CPs were separated by reverse-phase high-performance liquid chromatography. Twelve fractions were collected, and their ABTS free radical scavenging and ACE inhibitory activities were determined. Fractions 1, 2, 3, 4, and 9 showed higher ABTS free radical scavenging and ACE inhibitory activities. Twelve peptides were identified from active fractions by liquid chromatography–tandem mass spectrometry (LC–MS/MS) as Leu-Ser-Pro-Tyr, Leu-Asn-Ser-Pro-Ala-Tyr, Tyr-Gly-Pro-Gln, Pro-Pro-Tyr, Ala-Tyr-Pro-Glu, Thr-Tyr-Ser-Gly-Pro-Lys, Ala-Tyr-Pro-Gly-Pro-Gln, Leu-Ala-Tyr-Pro-Gln, Ala-Tyr-Leu-Gln-Gln-Gln, Ser-Ala-Pro, Asn-Ala-Pro, and Val-Asn-Ala-Pro. These peptides exhibited good ABTS free radical scavenging and ACE inhibitory activities. Therefore, CPs may have potential applications in the food industry as functional material additives.

BACKGROUND Food allergy has been regarded as a serious public nutritional health problem and attracted extensive attention all over the world, of which shellfish allergy is a long-lasting disorder that impacts health through a life-time. Sarcoplasmic calcium-binding protein (SCP) plays a vital role in cell and muscle functions and has been identified as an allergen in oyster. RESULTS In this study, recombinant SCP (rSCP) with a molecular mass of 21 kDa was produced and identified based on SCP amino acid sequence of pacific oyster (Crassostrea gigas), and was used as a follow-up experimental material. Its physicochemical characterization showed that the purified rSCP is highly stable to heat, acid-alkali and trypsin digestion, but weaker to pepsin digestion. We established an animal sensitization model and rSCP displayed stronger IgE-binding activity with rats serum in the rSCP+ CT group compared with the CT group and control group. Most importantly, five epitope peptides were identified as linear immunodominant epitopes by indirect competitive enzyme-linked immunosorbent assay (icELISA) for the first time. In addition, we also found that conformational epitopes may play a major role in immunoreactivity of SCP. CONCLUSION These results are significant for understanding hypersensitization of humans to oyster and offer available preventive measures and treatment programs in further research. This article is protected by copyright. All rights reserved.

Background Molluscan shellfish, including oysters, often cause allergic reactions in sensitive people throughout the world. It has been demonstrated that arginine kinase (AK) was one of the major allergens of oyster. The present study aimed to evaluate the immunoreactivity and structure of oyster arginine kinase (AK) as affected by heat treatment, pH change, and in vitro digestion. What's more, the IgE-binding epitopes of this allergen were also predicted and validated. Results Thermal and pH assays revealed that AK was unstable at temperature > 40°C or pH ≤5.0 by SDS-PAGE and Circular Dichroism (CD), while the digestibility assays suggested that AK was easier to be digested by pepsin than by trypsin and chymotrypsin. The potential epitopes were predicted through immunoinformatics tools and seven linear epitopes were identified by indirect competition enzyme-linked immunosorbent assay (icELISA) with pooled sera and individual serum from oyster allergic patients. The critical amino acids in each epitope were also confirmed using mutant peptides. These linear epitopes and critical amino acids were apt to distribute on the outer surface of homology-based AK model. Moreover, the three denaturants (SDS, β-Me and urea) can destroy the spatial structure of AK and increase or reduce its allergenicity by denaturation treatments. Conclusion Processing conditions lay the foundation for the variation of allergenicity. Seven linear epitopes and the critical amino acids of them were identified by indirect competitive enzyme-linked immunosorbent assay (icELISA). These findings will be helpful in allergy diagnosis and development of hypoallergenic products in the near future. This article is protected by copyright. All rights reserved.

Acute kidney injury (AKI) is a clinical syndrome that is defined as a sudden decline in renal function and characterized by inflammation and programmed cell death of renal tubular epithelial cells. Necroptosis is a form of regulated cell death that requires activation of receptor interacting protein kinase 3 (RIPK3) and its phosphorylation of the substrate MLKL. RIPK3 plays an important role in acute kidney injury, and hence developing its inhibitors is considered as one of the promising strategies aimed at prevention and treatment of AKI. Recently, we discovered AZD5423 as a novel potent RIPK3 inhibitor using a computer-aided hybrid virtual screening strategy according to three-dimensional structure of RIPK3. Our findings revealed that AZD5423 strongly inhibits activation of RIPK3, and MLKL phosphorylation upon cisplatin-, hypoxia/reoxygenation (H/R)- and TNF-α stimuli as compared with GSK872, which is a previously identified RIPK3 inhibitor. Importantly, AZD5423 exerts effective protection against cisplatin- and ischemia/reperfusion (I/R)-induced AKI mouse model. The results of cellular thermal shift assay and experiments in RIPK3 knockout cells indicated that AZD5423 could directly target RIPK3 to inhibit RIPK3 kinase activity. Mechanistically, the docking of AZD5423 and RIPK3 suggested that the kinase domain of RIPK3 for Lys

Zinc finger E-box-binding homeobox 1 (ZEB1), a functional protein of zinc finger family, was aberrant expressed in many kinds of liver disease including hepatic fibrosis and Hepatitis C virus. Bioinformatics results showed that ZEB1 was abnormally expressed in HCC tissues. However, to date, the potential regulatory role and molecular mechanisms of ZEB1 are still unclear in the occurrence and development of HCC. This study demonstrated that the expression level of ZEB1 was significantly elevated both in liver tissues of HCC patients and cell lines (HepG2 and SMMC-7721 cells). Moreover, ZEB1 could promote the proliferation, migration, and invasion of HCC cells. On the downstream regulation mechanism, ZEB1 could activate the Wnt/β-catenin signaling pathway by upregulating the protein expression levels of β-catenin, c-Myc, and cyclin D1. Novel studies showed that miR-708 particularly targeted ZEB1 3′-UTR regions and inhibited the HCC cell proliferation, migration, and invasion. Furthermore, results of nude mice experiments of HCC model indicated that miR-708 could inhibit tumor growth and xenograft metastasis model was established to validate that miR-708 could inhibit HCC cell metastasis through tail-vein injection in vivo. Together, the study suggested that ZEB1 modulated by miR-708 might be a potential therapeutic target for HCC therapy. [Image: see text]

In order to explore the mechanism underlying the ACE inhibitory activity of the active peptides derived from salmon skin in depth, salmon skin collagen peptides (SSCPs) were produced by the hydrolysis of Atlantic salmon (Salmo salar L.) skin and isolated by reverse-phase liquid chromatography. A total of 13 low-molecular-weight peptides were identified by mass spectrometry and used to analyze the inhibitory activity of angiotensin I–converting enzyme (ACE). Three peptides—Gly-Arg, Arg-Glu-Arg, and Gly-Pro-Arg—exhibited the highest ACE inhibitory activities with IC50 values of 0.73 ± 0.05, 0.89 ± 0.06, and 1.05 ± 0.13 mg/mL, respectively. These ACE inhibitory peptides were quantified, and their potential absorption, distribution, metabolism, excretion, and toxicity were predicted in silico together with two previously identified ACE inhibitory dipeptides Ala-Pro and Val-Arg. The molecular mechanism underlying the interaction between the peptides and ACE was determined using in silico methods, including molecular docking, pharmacophore modeling, and pharmacophore heat maps. The five ACE inhibitory peptides exhibited satisfactory absorption, distribution, metabolism, excretion, and toxicity. They formed hydrogen bonds, ionic bonds, metal–receptor bonds, and hydrophobic bonds with ACE to exert ACE inhibitory effects. Hydrogen bonding was the essential factor influencing ACE inhibitory activity.

For technical reasons, virtually all plastic deformation experiments on geological materials have been performed in either pure shear or simple shear. These special case loading geometries are rather restrictive for those seeking insight into how microstructure evolves under the more general loading geometries that occur during natural deformation. Moreover, they are insufficient to establish how plastic flow properties might vary with the 3rd invariant of the deviatoric stress tensor (J3) which describes the stress configuration, and so applications that use those flow properties (e.g. glaciological and geodynamical modelling) may be correspondingly compromised. We describe an inexpensive and relatively straightforward modification to the widely used Paterson rock deformation apparatus that allows torsion experiments to be performed under simultaneously applied axial loads. We illustrate the performance of this modification with the results of combined stress experiments performed on Carrara marble and Solnhofen limestone at 500°–600 °C and confining pressures of 300 MPa. The flow stresses are best described by the Drucker yield function which includes J3-dependence. However, that J3-dependence is small. Hence for these initially approximately isotropic calcite rocks, flow stresses are adequately described by the J3-independent von Mises yield criterion that is widely used in deformation modelling. Loading geometry does, however, have a profound influence on the type and rate of development of crystallographic preferred orientation, and hence of mechanical anisotropy. The apparatus modification extends the range of loading geometries that can be used to investigate microstructural evolution, as well as providing greater scope for determining the shape of the yield surface in plastically anisotropic materials.