Efficacy of Diosmin in Reducing Lower-Extremity Swelling and Pain After Total Knee Arthroplasty: A Randomized, Controlled Multicenter Trial.

BACKGROUND: Many patients experience lower-extremity swelling following total knee arthroplasty (TKA), which impedes recovery. Diosmin is a semisynthetic flavonoid that is often utilized to treat swelling and pain caused by chronic venous insufficiency. We aimed to evaluate the efficacy and safety of diosmin in reducing lower-extremity swelling and pain as well as in improving functional outcomes following TKA. METHODS: This study was designed as a randomized, controlled multicenter trial and conducted in 13 university-affiliated tertiary hospitals. A total of 330 patients undergoing TKA were randomized to either receive or not receive diosmin postoperatively. The diosmin group received 0.9 g of diosmin twice per day for 14 consecutive days starting on the day after surgery, whereas the control group received neither diosmin nor a placebo postoperatively. The primary outcome was lower-extremity swelling 1, 2, 3, and 14 days postoperatively. The secondary outcomes were postoperative pain assessed with use of a visual analogue scale, Hospital for Special Surgery score, range of knee motion, levels of the inflammatory biomarkers C-reactive protein and interleukin-6, and complications. RESULTS: At all postoperative time points, diosmin was associated with significantly less swelling of the calf, thigh, and upper pole of the patella as well as with significantly lower pain scores during motion. However, no significant differences in postoperative pain scores at rest, Hospital for Special Surgery scores, range of motion, levels of inflammatory biomarkers, or complication rates were found between the diosmin and control groups. CONCLUSIONS: The use of diosmin after TKA reduced lower-extremity swelling and pain during motion and was not associated with an increased incidence of short-term complications involving the outcomes studied. However, further studies are needed to continue exploring the efficacy and safety of diosmin use in TKA. LEVEL OF EVIDENCE: Therapeutic Level I . See Instructions for Authors for a complete description of levels of evidence.

Establishment and application of a VP3 antigenic domain-based peptide ELISA for the detection of antibody against goose plague virus infection.

The detection of antibody against goose plague virus (GPV) infection has never had a commercialized test kit, which has posed challenges to the prevention and control of this disease. In this study, bioinformatics software was used to analyze and predict the dominant antigenic regions of the main protective antigen VP3 of GPV. Three segments of bovine serum albumin (BSA) vector-coupled peptides were synthesized as ELISA coating antigens. Experimental results showed that the VP3-1 (358-392aa) peptide had the best reactivity and specificity. By using the BSA-VP3-1 peptide, a detection method for antibody against GPV infection was established, demonstrating excellent specificity with no cross-reactivity with common infectious goose pathogen antibodies. The intra-batch coefficient of variation and inter-batch coefficient of variation were both less than 7%, indicating good stability and repeatability. The dynamic antibody detection results of gosling vaccines and the testing of 120 clinical immune goose serum samples collectively demonstrate that BSA-VP3-1 peptide ELISA can be used to detect antibody against GPV in the immunized goose population and has higher sensitivity than traditional agar gel precipitation methods. Taken together, the developed peptide-ELISA based on VP3 358-392aa could be useful in laboratory viral diagnosis, routine surveillance in goose farms. The main application of the peptide-ELISA is to monitor the antibody level and vaccine efficacy for GPV, which will help the prevention and control of gosling plague.

Novel Dual-Target Kinase Inhibitors of EGFR and ALK Were Designed, Synthesized, and Induced Cell Apoptosis in Non-Small Cell Lung Cancer.

ALK-positive NSCLC coexisting with EGFR mutations is a frequently occurring clinical phenomenon. Targeting ALK and EGFR simultaneously may be an effective way to treat these cancer patients. In this study, we designed and synthesized ten new dual-target EGFR/ALK inhibitors. Among them, the optimal compound 9j exhibited good activity with IC50 values of 0.07829 ± 0.03 µM and 0.08183 ± 0.02 µM against H1975 (EGFR T790M/L858R) and H2228 (EML4-ALK) cells, respectively. Immunofluorescence assays indicated that the compound could simultaneously inhibit the expression of phosphorylated EGFR and ALK proteins. A kinase assay demonstrated that compound 9j could inhibit both EGFR and ALK kinases; thus, exerting an antitumor effect. Additionally, compound 9j induced apoptosis in a dose-dependent manner and inhibited the invasion and migration of tumor cells. All of these results indicate that 9j is worthy of further study.

Discovery of a potent EGFR and ALK dual mutation inhibitor containing N-(3-((4-((2-(cyclopropylsulfinyl)phenyl)amino)pyrimidin-2-yl)amino) phenyl)acrylamide scaffold.

A series of EGFR and ALK dual inhibitors containing sulfoxide and cyclopropyl groups were designed and synthesized. The lead compound 8a showed a significant activity against EGFR and ALK in both the enzymatic and cellular assays. The study of anti-tumor mechanism indicated that 8a could effectively block the phosphorylation of EGFR and ALK proteins, so as to effectively inhibiting the proliferation and inducing apoptosis of H1975 tumor cells, blocking the cell cycle and reducing the mitochondrial membrane potential inhibited the migration of H1975 cells. In vivo studies, compounds 8a and 8d can significantly subside the tumor tissue of nude mice without obvious toxicity.

Interface-engineering studies on the photoelectric properties and stability of the CsSnI3-SnS heterostructure.

The stability of Sn-based perovskites has always been the main obstacle to their application. Interface engineering is a very effective method for improving the stability of perovskites and the efficiency of batteries. Two-dimensional (2D) monolayer SnS is selected as a surface-covering layer for the CsSnI3 lead-free perovskite. The structure, electronic properties, and stability of the CsSnI3-SnS heterostructure are studied using density functional theory. Due to the different contact interfaces (SnI2 and CsI interfaces) of CsSnI3, the interface electronic-transmission characteristics are inconsistent in the CsSnI3-SnS heterostructure. Because of the difference in work functions, electrons flow at the interface of the heterostructure, forming a built-in electric field. The heterostructures form a type-I energy-level arrangement. Under the action of an electric field in the CsI-SnS heterostructure, electrons at the CsI interface recombine with holes at the SnS interface; however, the holes of the SnI2 interface and the electrons of the SnS interface are easily recombined in the SnI2-SnS heterostructure. Moreover, monolayer SnS can enhance the light absorption of the CsSnI3-SnS heterostructure. Monolayer SnS can inhibit the migration of iodine ions and effectively improve the structural stability of the SnI2-SnS interface heterostructure. This work provides a new theoretical basis for improving the stability of lead-free perovskites.

Biomechanics of the unilateral posterosuperior, unipedicular, and bipedicular approaches for treatment by percutaneous vertebroplasty: a comparative study.

Percutaneous vertebroplasty (PVP) via the unilateral posterosuperior approach has achieved good clinical results for the treatment of osteoporotic vertebral compression fractures. This study compared the biomechanical performance of a single vertebral body after PVP by the unilateral posterosuperior, unipedicular, and bipedicular approaches. Twenty-one vertebral bodies from the osteoporotic spine segments (T11-L1) of seven older female cadavers were randomly assigned to the unipedicular (group A), bipedicular (group B), or unilateral posterosuperior approach group (group C). After constructing the fracture compression model, PVP was performed by the different approaches. CT scans showed symmetrical, evenly distributed bone cement in groups B and C and unilaterally distributed cement in group A. The recovery rates of the anterior vertebral body height in groups B and C were significantly higher than those in group A after PVP (P<0.05). The left curvature elastic moduli after PVP were significantly higher in group A than in groups B and C; however, the right curvature moduli in group A were lower than in the other groups (P<0.05). The flexion, extension, and vertical compression elastic moduli were lowest in group B (P<0.05). After PVP, failure strength and stiffness in groups B and C were comparable (P>0.05) and higher than those in group A (P<0.05). PVP through the unilateral posterosuperior approach was superior to the unipedicular approach and comparable to the bipedicular approach based on the biomechanical performance of a single vertebral body. Due to its safety, simplicity, and efficacy, the unilateral posterosuperior approach is recommended for clinical application.

Synthesis and evaluation of new pirfenidone derivatives as anti-fibrosis agents.

Two series of new pirfenidone derivatives, in which phenyl groups or benzyl groups are attached to the nitrogen atom of the pyridin-2(1H)-one moiety were synthesized and evaluated as anti-fibrosis agents. Among them, compound 5d, with a (S)-2-(dimethylamino) propanamido group in the R2 position (series 1) exhibited 10 times the anti-fibrosis activity (IC50: 0.245 mM) of pirfenidone (IC50: 2.75 mM). Compound 9d (series 2) gave an IC50 of 0.035 mM against the human fibroblast cell line HFL1. The mechanism of the optimal compound inhibiting fibrosis was also studied.

Restricting the Formation of Pb-Pb Dimer via Surface Pb Site Passivation for Enhancing the Light Stability of Perovskite.

Poor light stability hinders the potential applications of perovskite optoelectronic devices. Recent experiments have demonstrated that the passivation surface via forming strong chemical bonds (SO4 -Pb, PO4 -Pb, Cl-Pb, O-Pb, and S-Pb) could effectively improve the light stability of perovskite solar cells. However, the underlying reasons are not clear. Herein, the elusive underlying mechanisms of light stability enhancement are explained in detail using first principles calculations. The small polaron model and self-trapped exciton model demonstrate that an iodine vacancy defect on the surface of perovskite could trap a free electron under light illumination, which leads to a significant rearrangement of the Pb-I lattice and creats a new chemical species, i.e., a Pb-Pb dimer bound in the typical perovskite of CH3 NH3 PbI3 . The Pb-Pb dimer distorts the Pb-I octahedral lattice and reduces the defect formation energy of the I atoms. The surface Pb site passivation can prevent the formation of the Pb-Pb dimer, thereby improving the light stability. In addition, the strong ionic bond could better stabilize the Pb site. The in-depth understanding of the light stability and the passivation mechanism in this study can promote the application of perovskite optoelectronic devices.

Circular RNA VMA21 ameliorates IL-1ß-engendered chondrocyte injury through the miR-495-3p/FBWX7 signaling axis.

This study explored the function of circular RNA VMA21 (circVMA21) in osteoarthritis (OA). IL-1ß inducement reduced the expression of circVMA21 in C28/I2 cells and human primary chondrocytes. Forced expression of circVMA21 heightened cell viability and attenuated cell apoptosis, accompanied by upregulation of Bcl-2, and downregulation of Bax and C-caspase-3 in C28/I2 cells in response to IL-1ß exposure. CircVMA21 overexpression diminished the expression of MMP1 and MMP13, augmented the expression of COL2A1, and impeded the production of IL-6, TNF-α, prostaglandin E2 (PGE2) and NO. CircVMA21 served as a competitive endogenous RNA by sponging miR-495-3p. F-box and WD40 domain protein 7 (FBWX7) was identified as a target of miR-495-3p. The compensation experiments affirmed that circVMA21-mediated protective effects on IL-1ß-irritated chondrocytes through the miR-495-3p/FBWX7 axis. The role of circVMA21 was also confirmed in an OA rat model. Collectively, these findings revealed a protective effect of circVMA21in OA by intercepting the miR-495-3p/FBWX7 crosstalk.

Intrauterine exposure of mice to arsenite induces abnormal and transgenerational glycometabolism.

The adverse, transgenerational effects on health caused by environmental pollutants are receiving increasing attention. For humans and mice, inorganic arsenic (iAs), a widespread environmental contaminant, is associated with diabetic phenotypes. However, the transgenerational effects of arsenite-induced changes in glucose metabolism in mice have not been fully investigated. In the present study, F0 pregnant mice were exposed to arsenite via drinking water (0, 0.5, 5, or 50 ppm NaAsO2) from gestational day 0 (GD0) until parturition. We examined the effects of arsenite exposure on the metabolic phenotypes and the levels of proteins and genes related to glucose metabolism of dams and their offspring (F1∼F4). Arsenite exposure altered the glucose tolerance of offspring. Notably, glucose transporter-2 (GLUT2) and insulin receptor substrate-1 (IRS1), which are related to the maintenance of glucose homeostasis, were also changed. The homeostasis assessment-insulin resistance (HOMA-IR), an indicator of insulin resistance, was higher in the offspring from the F0 female mice exposed to arsenite. Furthermore, imprinted genes, insulin-like growth factor 2 (IGF2) and potassium voltage-gated channel subfamily Q member 1 (KCNQ1), related to glycometabolism across multiple generations, were lower in the offspring. In sum, arsenite exposure during pregnancy transgenerationally affects glucose metabolism, which is related to altered levels of IGF2 and KCNQ1.

Hypoxia inducible factor-3α promotes osteosarcoma progression by activating KDM3A-mediated demethylation of SOX9.

Hypoxia/oxygen-sensing signally is closely associated with many tumor progressions, including osteosarcoma (OS). Previous research principally focused on the function of hypoxia-inducible factor (HIF)-1α and HIF-2α as the major hypoxia-associated transcription factors in OS, however, the role of HIF-3α has not been investigated. Our study found that HIF-3α was upregulated in OS tissues and cell lines. HIF-3α overexpression facilitated cell proliferation and invasion, and inhibited apoptosis, whereas HIF-3α knockdown showed the opposite results. Chromatin immunoprecipitation analysis revealed that lysine demethylase 3A (KDM3A) expression was transcriptionally activated by HIF-3α under hypoxia, and KDM3A occupied the SRY-box transcription factor 9 (SOX9) gene promoter region through H3 lysine 9 dimethylation (H3K9me2). Additionally, rescue results revealed that KDM3A or SOX9 overexpression reversed the effects of HIF-3α silence on cell functions. The Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) pathway inhibitor cucurbitacin I suppressed the promotive effects of HIF-3α overexpression on cell proliferation, invasion and TAK2/STAT3 pathway. Finally, OS cell line MG-63 transfected with HIF-3α short hairpin RNA (HIF-3α shRNA) were subcutaneously injected into nude mice, and the results found that HIF-3α knockdown significantly inhibited the xenograft tumor growth of OS in vivo. In conclusion, this study reveals that HIF-3α promotes OS progression in vitro and in vivo by activating KDM3A-mediated SOX9 promoter demethylation, which may provide a potential therapeutic mechanism for OS.

Polarization of rheumatoid macrophages is regulated by the CDKN2B-AS1/ MIR497/TXNIP axis.

The polarization of macrophages plays a critical role in the pathophysiology of rheumatoid arthritis. The macrophages can have pro-inflammatory M1 polarization and various types of alternative anti-inflammatory M2 polarization. Our preliminary results showed that the CDKN2B-AS1/MIR497/TXNIP axis might regulate macrophages of rheumatoid arthritis patients. Therefore, we hypothesized that this axis regulated the polarization of rheumatoid macrophages. Flow cytometry was used to determine the surface polarization markers in M1 or M2 macrophages from healthy donors and rheumatoid arthritis patients. The QPCR and Western Blotting were used to compare the expression of the CDKN2B-AS1/MIR497/TXNIP axis in these macrophages. We Knocked down and overexpressed the axis in the macrophage cell line MD to test its roles in macrophage polarization. Compared to cells from healthy donors, cells from rheumatoid arthritis patients expressed higher levels of CD40 and CD80 and lower levels of CD16, CD163, CD206, and CD200R after polarization, they also expressed higher CDKN2B-AS1, lower MIR497, and higher TXNIP. In macrophages from healthy donors, there was no correlation among CDKN2B-AS1, MIR497, and TXNIP. But in macrophages from patients, there were significant correlations. The CDKN2B-AS1 knockdown, MIR497 mimics suppressed the M1 polarization but promoted the M2 polarization in MD cells, while the MIR497 knockdown and the TXNIP overexpression did the opposite. This study demonstrated that elevated CDKN2B-AS1 in macrophages promotes the M1 polarization and inhibited the M2 polarization of macrophages by the CDKN2B-AS1/ MIR497/TXNIP axis.

In vitro and in vivo efficacy of the novel oral proteasome inhibitor NNU546 in multiple myeloma.

Proteasome inhibition demonstrates highly effective impact on multiple myeloma (MM) treatment. Here, we aimed to examine anti-tumor efficiency and underlying mechanisms of a novel well tolerated orally applicable proteasome inhibitor NNU546 and its hydrolyzed pharmacologically active form NNU219. NNU219 showed more selective inhibition to proteasome catalytic subunits and less off-target effect than bortezomib ex vivo. Moreover, intravenous and oral administration of either NNU219 or NNU546 led to more sustained pharmacodynamic inhibitions of proteasome activities compared with bortezomib. Importantly, NNU219 exhibited potential anti-MM activity in both MM cell lines and primary samples in vitro. The anti-MM activity of NNU219 was associated with induction of G2/M-phase arrest and apoptosis via activation of the caspase cascade and endoplasmic reticulum stress response. Significant growth-inhibitory effects of NNU219 and NNU546 were observed in 3 different human MM xenograft mouse models. Furthermore, such observation was even found in the presence of a bone marrow microenvironment. Taken together, these findings provided the basis for clinical trial of NNU546 to determine its potential as a candidate for MM treatment.

The ubiquitination and acetylation of histones are associated with male reproductive disorders induced by chronic exposure to arsenite.

Exposure to arsenic, which occurs via various routes, can cause reproductive toxicity. However, the mechanism for arsenic-induced reproductive disorders in male mice has not been extensively investigated. Here, 6-week-old male mice were dosed to 0, 5, 10, or 20 ppm sodium arsenite (NaAsO2), an active form of arsenic, in drinking water for six months. For male mice exposed to arsenite, fertility was lower compared to control mice. Moreover, for exposed mice, there were lower sperm counts, lower sperm motility, and higher sperm malformation ratios. Further, the mRNA and protein levels of the gonadotropin-regulated testicular RNA helicase (DDX25) and chromosome region maintenance-1 protein (CRM1), along with proteins associated with high mobility group box 2 (HMGB2), phosphoglycerate kinase 2 (PGK2), and testicular angiotensin-converting enzyme (tACE) were lower. Furthermore, chronic exposure to arsenite led to lower H2A ubiquitination (ubH2A); histone H3 acetylation K18 (H3AcK18); and histone H4 acetylations K5, K8, K12, and K16 (H4tetraAck) in haploid spermatids from testicular tissues. These alterations disrupted deposition of protamine 1 (Prm1) in testes. Overall, the present results indicate that the ubiquitination and acetylation of histones is involved in the spermiogenesis disorders caused by chronic exposure to arsenite, which points to a previously unknown connection between the modification of histones and arsenite-induced male reproductive toxicity.

Di-n-butyl phthalate promotes lipid accumulation via the miR200c-5p-ABCA1 pathway in THP-1 macrophages.

Di-n-butyl phthalate (DBP) is ubiquitously in the environment and has been detected in almost all of human bodies. Few data could be found about the effects of DBP on cardiovascular system, though its reproductive toxicities have been studied extensively. This study aimed to explore the effects of DBP on lipid metabolism, a key step during the formation of atherosclerosis, since DBP was recently reported to be associated with atherosclerosis. THP-1 macrophages were employed and exposed to various levels of DBP (10-8, 10-7, 10-6, 10-5 and 10-4 mol/L) or DMSO as control. Lipid accumulation was determined by detection of cellular total cholesterol, free cholesterol, cholesterol ester and content of lipid drops. Expressions of mRNA/miRNAs and proteins were measured by qRT-PCR and western blotting, respectively. Bioinformatic analysis and dual luciferase reporter assay were used to analyze the combination between miR200c-5p and ATP-binding cassette transporter A1 (ABCA1). Cholesterol efflux assay was executed to study the inhibitory effects of DBP on cholesterol efflux capability. Results revealed that DBP at 10-7 mol/L prompted THP-1 macrophages lipid accumulation by inhibiting cholesterol efflux via suppressing ABCA1 expression. In addition, a non-linear inverted U-shaped relationship between DBP and lipid accumulation could be observed. Moreover, miR200c-5p could directly targets to ABCA1 3'UTR and modulate ABCA1 expression. Besides, downregulation of ABCA1 expression and reduction of lipid efflux induced by DBP were due to the miR200c-5p upregulation. Collectively, these data suggested that DBP at levels relative to human exposure could increase lipid accumulation in THP-1 macrophages by decreasing cholesterol efflux through miR200c-5p-ABCA1, then potentiate the formation of atherosclerosis.

Multicomponent nanohybrids of nickel/ferric oxides and nickel cobaltate spinel derived from the MOF-on-MOF nanostructure as efficient scaffolds for sensitively determining insulin.

Multicomponent nanohybrids of nickel/ferric oxides and nickel cobaltate spinel (denoted as NiO/Fe2O3/NiCo2O4) have been prepared through pyrolyzing the hierarchical nanostructure of MOF-on-MOF and explored as efficient scaffolds for sensitively determining insulin. As for the MOF-on-MOF preparation, the ultra-thin bimetallic CoNi-zeolitic imidazolate framework (CoNi-ZIF) nanosheets were grown tightly around the bimetallic CoFe Prussian blue analogue (CoFe PBA) nanocube (denoted as CoNi-ZIF@CoFePBA). Basic characterizations revealed the original core-shell structure shape was still maintained in the NiO/Fe2O3/NiCo2O4 pyrolyzed at 300 °C, which was composed of multi-metal oxides and NiCo2O4 spinel, along with low crystallinity. Conversely, the NiO/Fe2O3/NiCo2O4 nanohybrid calcined at 600 °C consisted of large amounts of nanoparticles, while the nanohyrbid obtained at 900 °C demonstrated aggregated NiO and Fe2O3 nanoparticles coexisted with the NiCo2O4 phase. Owing to the porous nanostructure, the synergistic effect among different components, excellently electrochemical conductivity, and good biocompatibility of the NiO/Fe2O3/NiCo2O4 nanohybrid obtained at 600 °C, the relevant aptasensor displayed superior sensing performance for the determination of insulin. It gave an ultra-low detection limit of 9.1 fg mL-1 (0.16 fM) within a wide linear insulin concentration ranging from 0.01 pg mL-1 (0.172 fM) to 100 ng mL-1 (1.72 nM) determined by the electrochemical technique. The constructed aptasensor also had high selectivity, good stability, excellent reproducibility, and acceptable applicability in human serum. By integrating the advantages of aptasensors and electrochemical approach with features of multi-metallic metal-organic frameworks, this work widely broadens the applications of MOF-driven nanohybrids in biosensing fields.

Arsenite-induced transgenerational glycometabolism is associated with up-regulation of H3K4me2 via inhibiting spr-5 in caenorhabditis elegans.

Arsenic (As) is a toxic element that is highly abundant in the environment. However, there has not been sufficient research into the mechanisms of arsenic-induced transgenerational effects. In biomedical and environmental toxicology research field, C. elegans are often used as the ideal model. In this study, F0 generation animals were cultured with arsenite, while subsequent generations animals (F1 - F6) were cultured in the absence of arsenic. Experiments were performed to examine the transgenerational glycometabolism and the associated mechanisms in all seven generations (F0 - F6) of C. elegans. Results show that arsenite exposure increased total glucose content but reduced glucose metabolites in F0 generation C. elegans. The total glucose content was also elevated in subsequent generations probably due to transgenerational downregulation of fgt-1. In addition, arsenite exposure induced transgenerational downregulation of histone demethyltransferase spr-5 and elevated histone dimethylation in F0 generation. This study highlights that single generation exposure to arsenite causes transgenerational changes in glycometabolism in C. elegans, which may be caused by downregulation of spr-5 and elevation of H3K4me2.

Detection of Six ß-Agonists by Three Multiresidue Immunosensors Based on an Anti-bovine Serum Albumin-Ractopamine-Clenbuterol-Salbutamol Antibody.

According to an indirect competitive immunoassay, six ß-agonists (clenbuterol (CL), salbutamol (SAL), ractopamine (RAC), terbutaline (TER), mabuterol (MAB), and tulobuterol (TUL)) were detected by three novel multiresidue immunosensors on the basis of the successful preparation of bovine serum albumin (BSA)-RAC-CL-SAL multideterminant antigen and anti-BSA-RAC-CL-SAL antibody. A new strategy was reported to detect six ß-agonists by combining nanotechnology, electrochemical detection, and specific immune technology. At the same concentration, the amperometric response for detection of six ß-agonists was in a sequence of GCE/GNP/SAL > GCE/GNP/RAC > GCE/GNP/CL. Detection limits of six ß-agonists show that the multiresidue detection performance of the GCE/GNP/RAC immunosensor is better than those of GCE/GNP/SAL and GCE/GNP/CL immunosensors. Three immunosensors manifest superior properties with a wide linear range, low detection limit, excellent reproducibility, and stability. The proposed GCE/GNP/RAC immunosensor displays high accuracy and can be effectively used for real sample detection.

SREBP2-STARD4 is involved in synthesis of cholesteryl ester stimulated by mono-butyl phthalate in MLTC-1 cells.

Sterol is synthesized from cholesterol which is from the hydrolysis of stored cholesteryl esters. The process of maintaining cholesterol homeostasis is regulated by SREBP2-STARD4. Lots of researches demonstrated that male steroidogenesis could be interfered by di-n-butyl phthalate (DBP) or monobutyl phthalate (MBP). However, mechanisms of MBP exposure in this process have not been uncovered clearly. The objectiveof this study was to explore roles of SREBP2 and STARD4 in cholesteryl estersynthesis stimulated by MBP in mouse Leydig tumor cells (MLTC-1). MLTC-1 exposedto 10-8, 10-7, 10-6, 10-5 M MBP showed that levels of cholestery ester were increased significantly at 10-7 M MBP. Besides, cholesteryl ester synthesis stimulated by MBP was down-regulate when STARD4 or SREBP2 were inhibited. Activity of SREBP2 binding to the promoter of STARD4 was increased after MBP exposure. This study suggests that MBP can increase cholesteryl ester synthesis through SREBP2-STARD4 signal pathway in MLTC-1 cells.

A bimetallic (Cu-Co) Prussian Blue analogue loaded with gold nanoparticles for impedimetric aptasensing of ochratoxin a.

Bimetallic (Cu-Co) Prussian Blue analogs (PBAs) were coupled to gold nanoparticles to give a nanocomposite of type AuNP@CuCoPBA. It is shown to be a viable material for the impedimetric aptamer-based determination of ochratoxin A (OTA). Basic characterizations revealed that the chemical composition and crystal structure of AuNP@CuCoPBA is similar to that of pristine CuCo PBA. Nevertheless, the nanocube shape of CoCu PBA is converted to small nanoparticles on addition of AuNPs. Compared with CuCoPBA-based aptasensor, the AuNP@CuCoPBA-based assay exhibits excellent electrochemical conductivity, strong aptamer binding interaction, and high G-quadruplex stability. Electrochemical impedance spectroscopy revealed that the assay has limits of detection as low as 5.2 fg mL-1 of OTA, a response in the 50 fg mL-1 to 10 ng mL-1 concentration range, high selectivity, good reproducibility, repeatability, and acceptable applicability. In our perception, it represents a universal and powerful method for aptamer strand immobilization. It may be applied to the determination of various other analytes for which aptamers are available. Graphical abstract Schematic presentation of the preparation of a novel bimetallic Cu-Co PBAs coupling with gold nanoparticles (AuNPs) was supplied. The AuNP@CuCoPBA composite was applied for the sensitive detection of ochratoxin A (OTA). The impedimetric aptasensensor based on AuNP@CuCoPBA displays extremely low limits of detection toward OTA (5.2 fg mL-1), along with high selectivity, good reproducibility, high stability, repeatability, and acceptable applicability.