Pyrolysis-catalysis upcycling of waste plastic using a multilayer stainless-steel catalyst toward a circular economy.

Current un-sustainable plastic management is exacerbating plastic pollution, an urgent shift is thus needed to create a recycling society. Such recovering carbon (C) and hydrogen (H) from waste plastic has been considered as one practical route to achieve a circular economy. Here, we performed a simple pyrolysis-catalysis deconstruction of waste plastic via a monolithic multilayer stainless-steel mesh catalyst to produce multiwalled carbon nanotubes (MWCNTs) and H2, which are important carbon material and energy carrier to achieve sustainable development. Results revealed that the C and H recovery efficiencies were as high as 86% and 70%, respectively. The unique oxidation-reduction process and improvement of surface roughness led to efficient exposure of active sites, which increased MWCNTs by suppressing macromolecule hydrocarbons. The C recovery efficiency declined by only 5% after 10 cycles, proving the long-term employment of the catalyst. This catalyst can efficiently convert aromatics to MWCNTs by the vapor-solid-solid mechanism and demonstrate good universality in processing different kinds of waste plastics. The produced MWCNTs showed potential in applications of lithium-ion batteries and telecommunication. Owing to the economic profits and environmental benefits of the developed route, we highlighted its potential as a promising alternative to conventional incineration, simultaneously achieving the waste-to-resource strategy and circular economy.

A PilZ domain protein interacts with the transcriptional regulator HinK to regulate type VI secretion system in Pseudomonas aeruginosa.

Type VI secretion systems (T6SS) are bacterial macromolecular complexes that secrete effectors into target cells or the extracellular environment, leading to the demise of adjacent cells and providing a survival advantage. Although studies have shown that the T6SS in Pseudomonas aeruginosa is regulated by the Quorum Sensing system and second messenger c-di-GMP, the underlying molecular mechanism remains largely unknown. In this study, we discovered that the c-di-GMP-binding adaptor protein PA0012 has a repressive effect on the expression of the T6SS HSI-I genes in P. aeruginosa PAO1. To probe the mechanism by which PA0012 (renamed TssZ, Type Six Secretion System -associated PilZ protein) regulates the expression of HSI-I genes, we conducted yeast two-hybrid screening and identified HinK, a LasR-type transcriptional regulator, as the binding partner of TssZ. The protein-protein interaction between HinK and TssZ was confirmed through co-immunoprecipitation assays. Further analysis suggested that the HinK-TssZ interaction was weakened at high c-di-GMP concentrations, contrary to the current paradigm wherein c-di-GMP enhances the interaction between PilZ proteins and their partners. Electrophoretic mobility shift assays revealed that the non-c-di-GMP-binding mutant TssZR5A/R9A interacts directly with HinK and prevents it from binding to the promoter of the quorum-sensing regulator pqsR. The functional connection between TssZ and HinK is further supported by observations that TssZ and HinK impact the swarming motility, pyocyanin production, and T6SS-mediated bacterial killing activity of P. aeruginosa in a PqsR-dependent manner. Together, these results unveil a novel regulatory mechanism wherein TssZ functions as an inhibitor that interacts with HinK to control gene expression.

IGF2BP3 promotes glutamine metabolism of endometriosis by interacting with UCA1 to enhances the mRNA stability of GLS1.

BACKGROUND: Insulin like growth factor II mRNA binding protein 3 (IGF2BP3) has been implicated in numerous inflammatory and cancerous conditions. However, its precise molecular mechanisms in endometriosis (EMs) remains unclear. The aim of this study is to examine the influence of IGF2BP3 on the occurrence and progression of EMs and to elucidate its underlying molecular mechanism. METHODS: Efects of IGF2BP3 on endometriosis were confrmed in vitro and in vivo. Based on bioinformatics analysis, RNA immunoprecipitation (RIP), RNA pull-down assays and Fluorescent in situ hybridization (FISH) were used to show the association between IGF2BP3 and UCA1. Single-cell spatial transcriptomics analysis shows the expression distribution of glutaminase 1 (GLS1) mRNA in EMs. Study the effect on glutamine metabolism after ectopic endometriotic stromal cells (eESCs) were transfected with Sh-IGF2BP3 and Sh-UCA1 lentivirus. RESULTS: Immunohistochemical staining have revealed that IGF2BP3 was upregulated in ectopic endometriotic lesions (EC) compared to normal endometrial tissues (EN). The proliferation and migration ability of eESCs were greatly reduced by downregulating IGF2BP3. Additionally, IGF2BP3 has been observed to interact with urothelial carcinoma associated 1 (UCA1), leading to increased stability of GLS1 mRNA and subsequently enhancing glutamine metabolism. Results also demonstrated that IGF2BP3 directly interacts with the 3' UTR region of GLS1 mRNA, influencing its expression and stability. Furthermore, UCA1 was able to bind with c-MYC protein, stabilizing c-MYC mRNA and consequently enhancing GLS1 expression through transcriptional promotion. CONCLUSION: These discoveries underscored the critical involvement of IGF2BP3 in the elevation and stability of GLS1 mRNA in the context of glutamine metabolism by interacting with UCA1 in EMs. The implications of our study extended to the identification of possible therapeutic targets for individuals with EMs.

The Reduced Gray Matter Volume and Functional Connectivity of the Cerebellum in Type 2 Diabetes Mellitus with High Insulin Resistance.

INTRODUCTION: Insulin resistance is widely thought to be a critical feature in type 2 diabetes mellitus (T2DM), and there is significant evidence indicating a higher abundance of insulin receptors in the human cerebellum than cerebrum. However, the specific structural or functional changes in the cerebellum related to T2DM remain unclear, and the association between cerebellar alterations, insulin resistance, cognition, and emotion is yet to be determined. METHODS: We investigated neuropsychological performance, and structural and functional changes in specific cerebellar subregions in 43 T2DM patients with high insulin resistance (T2DM-highIR), 72 T2DM patients with low insulin resistance (T2DM-lowIR), and 50 controls. Furthermore, the correlation and stepwise multiple linear regression analysis were performed. RESULTS: Compared to the controls, T2DM exhibited lower cognitive scores and higher depressive/anxious scores. Furthermore, T2DM-highIR patients showed reduced gray matter volume (GMV) in the right cerebellar lobules VIIb, Crus I/II, and T2DM showed reduced GMV in left lobules I-IV compared to controls. Additionally, functional connectivity decrease was observed between the right lobules I-V and orbital part of the superior frontal gyrus in T2DM-highIR compared to both T2DM-lowIR and controls. Notably, there were negative correlations between the GMV of the lobules VIIb, Crus I/II, and updated homeostatic model assessment of insulin resistance, and positive correlation with executive/visuospatial performance in T2DM patients. CONCLUSIONS: These results suggest that the cerebellar lobules VIIb, Crus I/II, represent vulnerable brain regions in the context of insulin resistance. Overall, this study offers new insights into the neuropathophysiological mechanisms of brain impairment in patients with T2DM.

Segmental Abnormalities of White Matter Microstructure in Primary Hypothyroidism Identified by Automated Fiber Quantification.

INTRODUCTION: Hypothyroidism leads to impaired white matter (WM) integrity, associated with cognitive/neuropsychiatric dysfunction. However, the specific segmental abnormalities of the fibers remain unexplored. Therefore, this study aimed to investigate whether the damage of the WM is limited to a specific segment or the entire bundle via diffusion metrics using automated fiber quantification. METHODS: A cross-sectional study was conducted on 31 hypothyroid patients and 28 healthy controls. Thyroid-related hormone levels, cognitive/neuropsychiatric function, and diffusion tensor image data were collected and analyzed. Correlation and random forest analyses were also performed. RESULTS: The mean fractional anisotropy (FA) values were reduced at the fiber tract level. The mean diffusivity (MD), axial diffusivity (AD), and radial diffusivity (RD) values were increased in several fiber tracts, i.e., cingulum cingulate (CC), anterior forceps of corpus callosum (CCF_A). Significant correlations were found between cognitive function and diffusion indicators such as the FA value of the left corticospinal tract and arcuate fasciculus (AF), the MD value of left CC, the RD value of left AF, the AD value of left CC, and CCF_A. The widespread microstructure disruption was spread on multiple specific segments of different tracts at the point-wise level. The random forest revealed that the accuracy of recognizing hypothyroid patients was 82.5%, with the anterior component of CCF_A having the most significant contribution. CONCLUSION: WM microstructural integrity impairments were found in multi-segments of the multiple fiber bundles in hypothyroidism, which might be a potential mechanism of the underlying neurocognitive decline and cerebral impairment. The CCF_A might serve as a neuro biomarker for early warning of cerebral impairment in hypothyroidism.

Effect of lidocaine spray on reliving non-coring needle puncture-related pain in patients with totally implantable venous access port: a randomized controlled trial.

PURPOSE: Patients with the placement of a totally implantable venous access port (TIVAP) commonly suffer from pain caused by inserting a non-coring needle. At present, lidocaine cream and cold spray are extensively used for pain management, but they are complex to manage in busy medical environments and developing countries. The lidocaine spray combines the analgesic effect of lidocaine cream and the rapid onset of cold spray, which can effectively alleviate the pain related to non-coring needle puncture in patients with TIVAP. This randomized-controlled trial aimed to explore the effectiveness, acceptability, and safety of lidocaine spray in relieving the pain of non-coring needle puncture in patients with TIVAP. METHODS: A total of 84 patients who were hospitalized in the oncology department of a Grade III Level-A hospital in Shanghai from January 2023 to March 2023 and were implanted with TIVAP and required non-coring needle puncture were selected as the study subjects. The recruited patients were randomly assigned to the intervention group and the control group (n=42). Before routine maintenance, the intervention group received lidocaine spray 5 min before disinfection, while the control group received water spray 5 min before disinfection. The main clinical outcome was pain, and the degree of puncture pain in both groups was evaluated using the visual analogue scale. RESULTS: There were no significant differences between the two groups in age, gender, educational level, body mass index, port implantation time, and disease diagnosis (P>0.05). The pain score in the intervention and control groups was 15.12±6.61mm and 36.50±18.79mm, respectively (P<0.001). There were 2 (4.8%) patients with moderate pain in the intervention group and 18 (42.9%) patients with moderate pain in the control group (P<0.001). In the control group, 3 (7.1%) patients reported severe pain. The median comfortability score for the two groups of patients was 10, but there was a difference between the two groups (P<0.05) because the intervention group tilted to the right. The successful puncture rate of the first time puncture had no difference between the two groups, both being 100%. Moreover, 33 patients (78.6%) in the intervention group and 12 patients (28.6%) in the control group reported that they would choose the same spray for intervention in the future (P<0.001). During the 1 week of follow-up, 1 patient in the intervention group developed skin itching (P>0.05). CONCLUSIONS: The local use of lidocaine spray in patients with TIVAP is effective, acceptable, and safe to alleviate the pain caused by non-coring needle puncture. TRIAL REGISTRATION: Chinese Clinical Trial Registry (registration number: ChiCTR2300072976).

Effect of CD163 Modification on Glucocorticoid-Related Mortality in Patients with Hepatitis B Virus-Related Acute-on-Chronic Liver Failure.

Objective: This study aimed to assess the relationship between glucocorticoid treatment and mortality among patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF). Methods: We conducted a retrospective, hospital-based cohort study from 2019 to 2022, including 394 consecutively enrolled HBV-ACLF patients at the Third Affiliated Hospital of Chongqing Medical University. We recorded patient demographics, liver function, CD163 concentration, Model for End-Stage Liver Disease (MELD) score, and complications. The primary endpoint was 30-day mortality. Results: No significant differences were observed between the glucocorticoid-treated and non-glucocorticoid groups regarding sex, age, liver function, complications, or plasma CD163 concentration. After treatment, the median levels of total bilirubin (TBil), alanine aminotransferase (ALT), aspartate aminotransferase (AST), international normalized ratio (INR), and HBV DNA were 322.9 (IQR 258.6-383.3) µmol/L, 354.4 (IQR 253.1-444.6) U/L, 258.4 (IQR 186.4-322.4) U/L, 2.3 (IQR 2.1-2.5), and 5.0 (IQR 4.0-6.0) log IU/mL, respectively. Changes in ALT, AST, sCD163, TBil, INR, and MELD score before and after treatment showed no statistical differences between the glucocorticoid and non-glucocorticoid groups (P > .05). However, the mortality rate was significantly lower in the glucocorticoid group compared to the non-glucocorticoid group (11.2% vs. 29.9%, respectively; P < .001). Multivariable analysis revealed that, after adjusting for confounders, non-glucocorticoid treatment was associated with a higher adjusted hazard ratio (HR) for mortality (HR = 3.7, 95% CI 2.2-6.2) compared to glucocorticoid treatment. Additionally, an interaction test indicated that the association between non-glucocorticoid treatment and mortality was more robust in the sCD163 ≥ 18.2 mg/L group (HR = 7.6, 95% CI 2.9-19.9) but weaker in the sCD163 < 18.2 mg/L group (HR = 2.2, 95% CI 1.2-4.3) (P for interaction < .05). Conclusions: These findings suggest that glucocorticoids are an effective treatment for reducing mortality in HBV-ACLF patients, with particular effectiveness observed in patients with high sCD163 concentrations.

Polysaccharide H-1-2 Ameliorates High Glucose-Induced Podocyte Dysfunction by Suppressing Epithelial-to-Mesenchymal Transition via Restoration of SIRT1 in Vivo and in Vitro.

Renal interstitial fibrosis, a pathological feature of diabetic nephropathy, is closely related to endothelial-to-mesenchymal transition (EMT). This study aimed to explore the effect of H-1-2, a polysaccharide of Pseudostellaria heterophylla, on high glucose (HG) induced-podocyte EMT in vivo and ex vivo. DBA/2 mice were given five consecutive days of streptozotocin injection to induce the diabetic nephropathy model. H-1-2 treatment effectively attenuated general states (bodyweight and blood glucose level) and reduced oral glucose tolerance, insulin tolerance, kidney index, as well as the level of serum urine nitrogen, serum creatinine, and urinary albumin excretion rate in diabetic nephropathy mice. The injury and EMT of podocytes in diabetic nephropathy mice were restrained by H-1-2. After exposing podocytes to HG, the impaired cell viability, apoptosis, the downregulation of nephrin, synaptopodin, sirtuin 1 (SIRT1) and P-cadherin, and the upregulation of N-cadherin were observed in podocytes. H-1-2 treatment could reverse these effects induced by HG. To uncover the mechanism underlying H-1-2 suppressing EMT, small interference RNA for SIRT1 was transfected into podocytes. Mechanically, silencing SIRT1 largely restrained the protective effect of H-1-2 on HG-induced podocytes. In conclusion, H-1-2 exerts a potential role in alleviating HG-induced dysfunction and EMT of podocytes in vivo and ex vivo via SIRT1.

Improved Dynamic Event-Triggered Robust Control for Flexible Robotic Arm Systems with Semi-Markov Jump Process.

In this paper, we investigate the problem of a dynamic event-triggered robust controller design for flexible robotic arm systems with continuous-time phase-type semi-Markov jump process. In particular, the change in moment of inertia is first considered in the flexible robotic arm system, which is necessary for ensuring the security and stability control of special robots employed under special circumstances, such as surgical robots and assisted-living robots which have strict lightweight requirements. To handle this problem, a semi-Markov chain is conducted to model this process. Furthermore, the dynamic event-triggered scheme is used to solve the problem of limited bandwidth in the network transmission environment, while considering the impact of DoS attacks. With regard to the challenging circumstances and negative elements previously mentioned, the adequate criteria for the existence of the resilient H∞ controller are obtained using the Lyapunov function approach, and the controller gains, Lyapunov parameters and event-triggered parameters are co-designed. Finally, the effectiveness of the designed controller is demonstrated via numerical simulation using the LMI toolbox in MATLAB.

The Effect of Epimedium Isopentenyl Flavonoids on the Broiler Gut Health Using Microbiomic and Metabolomic Analyses.

Epimedium (EM), also known as barrenwort, is a traditional medicinal plant rich in isopentenyl flavonols, which have beneficial biological activities and can improve human and animal health, but its mechanism is still unclear. In this study, ultra-high-performance liquid chromatography/quadrupole-time-of-flight-mass spectrometry (UHPLC-Q-TOF/MS) and ultra-high-performance liquid chromatography triple-quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) were used to analyse the main components of EM, and isopentenyl flavonols such as Epimedin A, B, and C as well as Icariin were the major components of EM. Meanwhile, broilers were selected as model animals to illuminate the mechanism of Epimedium isopentenyl flavonols (EMIE) on gut health. The results showed that supplementation with 200 mg/kg EM improved the immune response, increased cecum short-chain fatty acids (SCFAs) and lactate concentrations, and improved nutrient digestibility in broilers. In addition, 16S rRNA sequencing showed that EMIE altered the composition of cecal microbiome, increasing the relative abundance of beneficial bacteria (Candidatus Soleaferrea and Lachbospiraceae NC2004 group and Butyricioccus) and reducing that of harmful bacteria (UBA1819, Negativibacillus, and Eisenbergiella). Metabolomic analysis identified 48 differential metabolites, of which Erosnin and Tyrosyl-Tryptophan were identified as core biomarkers. Erosnin and tyrosyl-tryptophan are potential biomarkers to evaluate the effects of EMIE. This shows that EMIE may regulate the cecum microbiota through Butyricicoccus, with changes in the relative abundance of the genera Eisenbergiella and Un. Peptostreptococcaceae affecting the serum metabolite levels of the host. EMIE is an excellent health product, and dietary isopentenyl flavonols, as bioactive components, can improve health by altering the microbiota structure and the plasma metabolite profiles. This study provides the scientific basis for the future application of EM in diets.

Effects of Tigecycline combined with Cefoperazone on bacterial clearance and serum biochemical indexes in patients with pulmonary infections in ICU.

Objectives: To investigate the effects of tigecycline combined with Cefoperazone on bacterial clearance and the expression of serum biochemical indexes [C-reactive protein (CRP), leukocyte count (WBC) and procalcitonin (PCT)] in patients with infection in an intensive care unit (ICU). Methods: The clinical data of 79 patients with pulmonary infections within the ICU of Chenzhou first people's Hospital from October 2019 to September 2021 were retrospectively analyzed. From the total, 38 patients received intravenous drip of Cefoperazone (control group), and 41 patients received intravenous drip of Cefoperazone and tigecycline (observation group). The treatment effect, bacterial clearance effect, serum biochemical index level and adverse reactions of the two groups were counted before and after treatment. Results: The total efficacy in the observation group (95.12%) was higher than that of the control group (78.95%) (P<0.05). After treatment, the bacterial clearance rate in the observation group (87.04%) was higher than that in the control group (66.67%) (P<0.05). After treatment, the levels of CRP, WBC and PCT in the two groups were lower than those before treatment (P<0.05), and the levels in the observation group were lower than those in the control group (P<0.05). There was no significant difference in the incidence of adverse reactions between the observation group (9.76%) and the control group (5.26%) (P>0.05). Conclusions: The combination of Cefoperazone and tigecycline in the treatment of ICU infection can effectively improve the treatment effect of the disease, have a significant bacterial clearance effect, and can reduce the serum levels of CRP, WBC and PCT.

Fluorescent probes based on nucleophilic aromatic substitution reactions for reactive sulfur and selenium species: Recent progress, applications, and design strategies.

Reactive sulfur species (RSS) and reactive selenium species (RSeS) are important substances for the maintenance of physiological balance. Imbalance of RSS and RSeS is closely related to a series of human diseases, so it is considered to be an important biomarker in early diagnosis, treatment, and stage monitoring. Fast and accurate quantitative analysis of different RSS and RSeS in complex biological systems may promote the development of personalized diagnosis and treatment in the future. One way to explore the physiological function of various types of RSS and RSeS in vivo is to detect them at the molecular level, and one of the most effective methods for this is to use fluorescent probes. Nucleophilic aromatic substitution (SNAr) reactions are commonly exploited as a detection mechanism for RSS and RSeS in fluorescent probes. In this review, we cover recent progress in fluorescent probes for RSS and RSeS based on SNAr reactions, and discuss their response mechanisms, properties, and applications. Benzenesulfonate, phenyl-O ether, phenyl-S ether, phenyl-Se ether, 7-nitro-2,1,3-benzoxadiazole (NBD), benzoate, and selenium-nitrogen bonds are all good detection groups. Moreover, based on an integration of different reports, we propose the design and synthesis of RSS- and RSeS-selective probes based on SNAr reactions, current challenges, and future research directions, considering the selection of active sites, the effect of substituents on the benzene ring, and the introduction of other functional groups.

Chinese pediatric Tuina on children with acute diarrhea: a randomized sham-controlled trial.

BACKGROUND: Pediatric Tuina has been widely used in children with acute diarrhea in China. However, due to the lack of high-quality clinical evidence, the benefit of Tuina as a therapy is not clear. We aimed to assess the effect of pediatric Tuina compared with sham Tuina as an add-on therapy in addition to usual care for 0-6-year-old children with acute diarrhea. METHODS: Eighty-six participants aged 0-6 years with acute diarrhea were randomized to receive pediatric Tuina plus usual care (n = 43) or sham Tuina plus usual care (n = 43). The primary outcomes were days of diarrhea from baseline and times of diarrhea on day 3. Secondary outcomes included a global change rating (GCR) and the number of days when the stool characteristics returned to normal. Adverse events were assessed. RESULTS: Pediatric Tuina was associated with a reduction in times of diarrhea on day 3 compared with sham Tuina in both ITT (crude RR, 0.73 [95% CI, 0.59-0.91]) and PP analyses (crude RR, 0.66 [95% CI, 0.53-0.83]). However, the results were not significant when we adjusted for social demographic and clinical characteristics. No significant difference was found between groups in days of diarrhea, global change rating, or number of days when the stool characteristics returned to normal. CONCLUSIONS: In children aged 0-6 years with acute diarrhea, pediatric Tuina showed significant effects in terms of reducing times of diarrhea compared with sham Tuina. Studies with larger sample sizes and adjusted trial designs are warranted to further evaluate the effect of pediatric Tuina therapy. TRIAL REGISTRATION: Clinicaltrials.gov, Identifier: NCT03005821 , Data of registration: 2016-12-29.

Optimized Golay-9 array configurations for mid-frequency compensation in optical sparse aperture systems.

Optical sparse aperture (OSA) imaging systems show great potential to generate higher resolution images than those of equivalent single filled aperture systems. However, due to the sparsity and dispersion of sparse aperture arrays, pupil function is no longer a connected domain, which further attenuates or loses the mid-frequency modulation transfer function (MTF), resulting in lower mid-frequency contrast and blurred images. Therefore, an improved traversal algorithm is proposed to optimize Golay-9 array configurations for compensating the mid-frequency MTF. Its structural parameters include diameters of sub-apertures, relative rotation angles between individual sub-apertures, and radius of concentric circles. Then, these parameters are traversed successively in order. Finally, the influences of the obtained optimized array configurations on the mid-frequency MTF are analyzed in detail, and the image performances are evaluated. The experimental results prove the contrast enhancement. Compared with a Golay-9 array at F=36.5%, the maximum MTF increases from 0.1503 to 0.307, and the mid-frequency MTF is boosted from 0.0565 to 0.0767. In addition, the peak signal to noise ratio of the degraded image is promoted from 19.75 dB to 20.63 dB. Both quantitative and qualitative evaluations demonstrate the validity of the proposed method.

Synergistic effects of D-arginine, D-methionine and D-histidine against Porphyromonas gingivalis biofilms.

Porphyromonas gingivalis biofilms are implicated in the pathology of peri-implantitis and periodontitis. In this study, D-arginine (R), D-methionine (M), D-histidine (H), and a mixture of these D-amino acids (D-AAs) were investigated as an effective therapeutic strategy against P. gingivalis biofilms. The bacterial growth activity and minimum inhibitory concentrations were determined for each D-AA, along with the effects of the D-AAs mixture on biofilm development, morphology, structure, extracellular polysaccharides (EPS), cytotoxicity towards commensals, and bacterial structure. The D-AA mixture delayed the proliferation of P. gingivalis, changed its membrane structure, and decreased biofilm thickness and integrity, as compared with individual D-AAs. The EPS content increased with the concentration of D-AAs. The present study shows that a 4 mM RMH, triple D-AA mixture, enhanced deleterious effects on P. gingivalis biofilms without any cytotoxicity compared with individual D-AAs, thus providing a new strategy for the treatment of peri-implantitis and periodontitis.

Isoorientin plays an important role in alleviating Cadmium-induced DNA damage and G0/G1 cell cycle arrest.

Cadmium is a heavy metal pollutant that has been reported to cause oxidative stress, apoptosis, and autophagy in cells, while the flavone isoorientin is a traditional Chinese medicine extract that has proven antioxidant and anti-inflammatory properties. Accordingly, in this study we used the rat proximal tubular cell line NRK-52E and primary rat proximal tubular (rPT) cells as models to investigate the effects of isoorientin against Cadmium-induced cell injury and the mechanism of these effects. Comet assay, Western blot, flow cytometry, immunofluorescence, and transmission electron microscopy were used to evaluate cell damage and cell-cycle-related protein expression. Furthermore, real-time cell analysis, cell-counting kit-8, and ELISA were used to investigate the role of isoorientin in Cadmium-induced cell injury. The results revealed that treatment of rat renal tubular epithelial cells with 2.5 µM Cd for 12 h resulted in DNA damage and G0/G1 cell cycle arrest, while isoorientin attenuated this Cd-induced damage.

Effect of oleic acid on induction of steatosis and cytotoxicity in BRL 3A cells.

Recent studies have shown that monounsaturated oleic acid induces steatosis in cultured hepatocyte steatosis in the form of nonalcoholic fatty liver disease models in vitro. However, the underlying mechanism of steatosis development is not completely understood. Therefore, we investigated the molecular mechanism of steatosis and the role of mitogen-activated protein kinase (MAPK)/toll-like receptor 4-related protein (TLR4) expression in this study. Rat hepatocyte cells were subjected to oleic acid in different concentrations (1.2-2.4 mM) for 24 hours. The cell morphological injury index and the changes in the MAPK/TLR4 signaling pathway-related proteins were evaluated. We found that the microstructure of the cells in the oleic acid treatment group was damaged, and higher phosphorylation levels of the MAPK pathway-related proteins were detected than those in the control group. In addition, the protein expression of TLR4, sterol regulatory element-binding protein-1, and fatty acid synthase were increased in the oleic acid treatment group. Our findings demonstrate that oleic acid causes toxic damage to rat hepatocyte cells, and the MAPK/TLR4 signaling pathway plays a significant role in lipid storage.

Comparison of soybean peroxidase with horseradish peroxidase and alkaline phosphatase used in immunoassays.

Enzyme labeling of an antigen or an antibody helps to visualize and amplify the signal and is an important reagent used in immunoassays for the detection of a target of interest. In this research, soybean peroxidase (SBP), a less commonly used enzyme reporter, was compared in immunoassays with the two most commonly used reagents, horseradish peroxidase (HRP) and alkaline phosphatase (ALP). The enzyme-antibody conjugates were evaluated by their performance in an indirect competitive enzyme-linked immunosorbent assay (icELISA) and in an indirect competitive chemiluminescent enzyme immunoassay (icCLEIA) for ractopamine (RAC). The results revealed that the more affordable SBP offers a long-lasting chemiluminescent signal, which outperformed ALP and HRP. SBP-antibody conjugate (SBP-Ab) based immunoassays produced lower limits of detection (LODs) and better accuracy in the detection of RAC in animal urine samples. Additionally, SBP-Ab has advantages in being more resistant to heat, acid and organic solvents. These results suggest that SBP could be a potentially excellent alternative to HRP and ALP for the development of immunoassay in food safety field.

MicroRNA-224 enhances the osteoblastic differentiation of hMSCs via Rac1.

Osteogenesis is the differentiation of mesenchymal stem cells (MSCs) into osteoblasts. MicroRNAs (miRNAs) are short noncoding RNAs that target specific genes to mediate translational activities. In this study, we investigated how miR-224 regulates the osteoblastic differentiation of human MSCs (hMSCs) as well as the underlying mechanism. The results revealed the upregulation of miR-224 during hMSC differentiation. In vitro experiments showed that the downregulation of miR-224 suppressed the differentiation of hMSCs into osteoblasts. However, upregulation of miR-224 was concomitant with increased expression of relevant genes and augmented activity of alkaline phosphatase. Furthermore, the results indicated that Rac1 acted as the bona fide target of miR-224 and that Rac1 depletion promoted osteogenic differentiation in miR-224-silenced hMSCs. In addition, we found that both JAK/STAT3 and Wnt/ß-catenin pathways were repressed by Rac1 depletion using quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blotting, and immunofluorescence. Our data indicate a novel molecular mechanism in relation to hMSCs differentiation into osteoblasts, which may facilitate bone anabolism via miR-224. SIGNIFICANCE OF THE STUDY: In this study, we mainly explored the effects of miR-224 on hMSCs differentiation into osteoblasts. We find that induced miR-224 expression in hMSCs is considered closely associated with specific osteogenesis-related genes, alkaline phosphatase activity, and matrix mineralization, indicating that miR-224 may serve as a promising biomarker for osteogenic differentiation. Our data indicate a novel molecular mechanism in relation to hMSCs differentiation into osteoblasts, which may facilitate bone anabolism via miR-224.

Metabolic Profile, Bioavailability and Toxicokinetics of Zearalenone-14-Glucoside in Rats after Oral and Intravenous Administration by Liquid Chromatography High-Resolution Mass Spectrometry and Tandem Mass Spectrometry.

Zearalenone-14-glucoside (ZEN-14G), a key modified mycotoxin, has attracted a great deal of attention due to the possible conversion to its free form of zearalenone (ZEN) exerting toxicity. In this study, the toxicokinetics of ZEN-14G were investigated in rats after oral and intravenous administration. The plasma concentrations of ZEN-14G and its major five metabolites were quantified using a validated liquid chromatography tandem mass spectrometry (LC-MS/MS) method. The data were analyzed via non-compartmental analysis using software WinNonlin 6.3. The results indicated that ZEN-14G was rapidly hydrolyzed into ZEN in vivo. In addition, the major parameters of ZEN-14G following intravenous administration were: area under the plasma concentration-time curve (AUC), 1.80 h·ng/mL; the apparent volume of distribution (VZ), 7.25 L/kg; and total body clearance (CL), 5.02 mL/h/kg, respectively. After oral administration, the typical parameters were: AUC, 0.16 h·ng/mL; VZ, 6.24 mL/kg; and CL, 4.50 mL/h/kg, respectively. The absolute oral bioavailability of ZEN-14G in rats was about 9%, since low levels of ZEN-14G were detected in plasma, which might be attributed to its extensive metabolism. Therefore, liquid chromatography high-resolution mass spectrometry (LC-HRMS) was adopted to clarify the metabolic profile of ZEN-14G in rats' plasma. As a result, eight metabolites were identified in which ZEN-14-glucuronic acid (ZEN-14GlcA) had a large yield from the first time-point and continued accumulating after oral administration, indicating that ZEN-14-glucuronic acid could serve a potential biomarker of ZEN-14G. The obtained outcomes would prompt the accurate safety evaluation of ZEN-14G.