Silencing PinX1 enhances radiosensitivity and antitumor-immunity of radiotherapy in non-small cell lung cancer.

BACKGROUND: We aimed to investigate the effects of PinX1 on non-small cell lung cancer(NSCLC) radiosensitivity and radiotherapy-associated tumor immune microenvironment and its mechanisms. METHODS: The effect of PinX1 silencing on radiosensitivity in NSCLC was assessed by colony formation and CCK8 assay, immunofluorescence detection of γ- H2AX and micronucleus assay. Western blot was used to assess the effect of PinX1 silencing on DNA damage repair pathway and cGAS-STING pathway. The nude mouse and Lewis lung cancer mouse model were used to assess the combined efficacy of PinX1 silencing and radiotherapy in vivo. Changes in the tumor immune microenvironment were assessed by flow cytometry for different treatment modalities in the Lewis luuse model. The interaction protein RBM10 was screened by immunoprecipitation-mass spectrometry. RESULTS: Silencing PinX1 enhanced radiosensitivity and activation of the cGAS-STING pathway while attenuating the DNA damage repair pathway. Silencing PinX1 further increases radiotherapy-stimulated CD8+ T cell infiltration and activation, enhances tumor control and improves survival in vivo; Moreover, PinX1 downregulation improves the anti-tumor efficacy of radioimmunotherapy, increases radioimmune-stimulated CD8+ T cell infiltration, and reprograms M2-type macrophages into M1-type macrophages in tumor tissues. The interaction of PinX1 and RBM10 may promote telomere maintenance by assisting telomerase localization to telomeres, thereby inhibiting the immunostimulatory effects of IR. CONCLUSIONS: In NSCLC, silencing PinX1 significantly contributed to the radiosensitivity and promoted the efficacy of radioimmunotherapy. Mechanistically, PinX1 may regulate the transport of telomerase to telomeres through interacting with RBM10, which promotes telomere maintenance and DNA stabilization. Our findings reveal that PinX1 is a potential target to enhance the efficacy of radioimmunotherapy in NSCLC patients.

BIBR1532 combined with radiotherapy induces ferroptosis in NSCLC cells and activates cGAS-STING pathway to promote anti-tumor immunity.

BACKGROUND: Telomerase, by safeguarding damaged telomeres and bolstering DNA damage repair, has the capacity to heighten the radioresistance of tumour cells. Thus, in turn, can compromise the efficacy of radiotherapy (RT) and radioimmunotherapy. Our previous studies have revealed that the highly selective telomerase inhibitor, BIBR1532, possesses the potential to enhance the radiosensitivity of Non-small cell lung cancer (NSCLC). In this study, we delve further into the impact of BIBR1532 on the immune activation induced by RT and elucidate the underlying mechanisms. METHODS: Biological information analyses, immunofluorescence assays, western blot assays, flow cytometry analysis were conducted to elucidate the functions of the combination of BIBR1532 with radiotherapy in NSCLC. Intracellular levels of lipid peroxides, glutathione, malondialdehyde, and Fe2+ were measured as indicators of ferroptosis status. Both in vitro and in vivo studies were conducted to examine the antitumor effects. RESULTS: Our findings indicate that the confluence of BIBR1532 with RT significantly augments the activation of the cGAS-STING pathway in both in vivo and in vitro settings, thereby fostering an effective anti-tumoral immune response. The effects can be ascribed to two key processes. Firstly, ionizing radiation, in precipitating DNA double-strand breaks (DSBs), prompts the release of tumour-derived double-stranded DNA (dsDNA) into the cytoplasm. Subsequently, BIBR1532 amplifies the activation of antigen-presenting cells by dsDNA post-RT and instigates the cGAS-STING pathway. Secondly, BIBR1532 enhances the ferroptosis response in NSCLC following RT, thereby promoting unrestrained lipid peroxidation and elevated levels of reactive oxygen species (ROS) within tumour cells. This ultimately leads to mitochondrial stress and the release of endogenous mitochondrial DNA (mtDNA) into the cytoplasm, thus facilitating the activation of the STING pathway and the induction of a type I interferon (IFN)-linked adaptive immune response. CONCLUSION: This study underscores the potential of BIBR1532 as an efficacious and safe radiosensitizer and radioimmunotherapy synergist, providing robust preclinical research evidence for the treatment of NSCLC.

miR-125a-3p regulates the expression of FSTL1, a pro-inflammatory factor, during adipogenic differentiation, and inhibits adipogenesis in mice.

Adipogenesis is tightly regulated by various factors, including genes and microRNAs. Excessive fat deposition is the key feature of obesity, which is a low-grade chronic inflammatory disease. Follistatin-like 1 (FSTL1) has been reported to be an important mediator involved in various inflammatory diseases. However, the underlying mechanism of FSTL1 in preadipocyte differentiation and inflammatory response is still unclear. The current study was designed to explore the biological function and potential mechanism of FSTL1 in mouse subcutaneous preadipocyte differentiation. We found that FSTL1 was highly expressed in the early stage of differentiation and subsequently decreased sharply, suggesting that FSTL1 played a possible role in adipogenesis. Meanwhile, the gain- and loss-of-function assays showed that FSTL1 was not only involved in the inflammatory response by inducing the expression of pro-inflammatory factors IL-1ß and CCL2 but also significantly attenuated preadipocyte differentiation, as evidenced by the reduction of lipid accumulation and the levels of adipogenic genes, including PPARγ and FABP4. In addition, the target gene prediction and luciferase reporter assay validated that miR-125a-3p targeted the 3' UTR region of FSTL1. These results demonstrated that miR-125a-3p negatively regulated the expression of FSTL1 at the mRNA and protein levels. Furthermore, overexpressing miR-125a-3p in preadipocytes dramatically accelerated adipogenic differentiation and downregulated the levels of IL-1ß and CCL2, which were in accordance with the knockdown of FSTL1. On the contrary, treatment with miR-125a-3p inhibitors attenuated adipogenesis but induced the expression of inflammatory genes. In summary, this study suggests a positive function of FSTL1 in adipocyte-induced inflammation and negatively regulates preadipocyte differentiation. Further studies demonstrated that miR-125a-3p could reverse the effect by targeting FSTL1, which might provide a better understanding of treating obesity-related inflammatory diseases.

LncRNA H19 Influences Cellular Activities via the miR-454-3p/BHLHE40 Axis in Anaplastic Thyroid Carcinoma.

Anaplastic thyroid carcinoma (ATC) is an aggressive malignancy threatening patients' life quality. Our previous study has demonstrated that inhibition of long non-coding RNA H19 (lncRNA h19; H19) blocks ATC growth and metastasis. The current study aimed to further explore the potential mechanism of H19 in ATC. Expression of H19, miR-454-3p, and BHLHE40 mRNA was measured using RT-qPCR in tissue samples and cell lines. The dual-luciferase reporter assay and Pearson correlation analysis were used to explore the interaction among H19, miR-454-3p, and BHLHE40. The biological process of proliferation, migration, and invasion was determined using loss- or gain-function CCK-8 and Transwell assays. Western blot assay was used to evaluate the changes in protein levels. H19 was elevated in ATC tissues and cell lines. Based on online prediction database results, miR-454-3p might be a target of H19, and BHLHE40 might be a direct target of miR-454-3p. miR-454-3p expression was decreased in ATC and had a negative interaction with H19. BHLHE40 mRNA expression was increased and has a negative correlation with miR-454-3p and a positive correlation with H19. Downregulation of miR-454-3p and upregulation of BHLHE40 could reverse the decreased cellular activities caused by si-H19. Moreover, the silence of H19 modulates BHLHE40 to affect the PI3K/AKT protein levels and apoptotic-related protein levels. The current study provided a potential detailed mechanism of H19 in ATC, and lncRNA H19-miR-454-3p-BHLHE40 interaction may be a new experimental basis for prognosis and targeted therapy for ATC patients.

Silencing of circIgf1r plays a protective role in neuronal injury via regulating astrocyte polarization during epilepsy.

Epilepsy is a common brain disorder, repeated seizures of epilepsy may lead to a series of brain pathological changes such as neuronal or glial damage. However, whether circular RNAs are involved in neuronal injury during epilepsy is not fully understood. Here, we screened circIgf1r in the status epilepticus model through circRNA sequencing, and found that it was upregulated after the status epilepticus model through QPCR analysis. Astrocytes polarizing toward neurotoxic A1 phenotype and neurons loss were observed after status epilepticus. Through injecting circIgf1r siRNA into the lateral ventricle, it was found that knocking down circIgf1r in vivo would induce the polarization of astrocytes to phenotype A2 and reduce neuronal loss. The results in vitro further confirmed that inhibiting the expression of circIgf1r in astrocytes could protect neurons by converting reactive astrocytes from A1 to the protective A2. In addition, knocking down circIgf1r in astrocytes could functionally promote astrocyte autophagy and relieve the destruction of 4-AP-induced autophagy flux. In terms of mechanism, circIgf1r promoted the polarization of astrocytes to phenotype A1 by inhibiting autophagy. Taken together, our results reveal circIgf1r may serve as a potential target for the prevention and treatment of neuron damage after epilepsy.

Poly(N,N-dimethyl)acrylamide-based ion-conductive gel with transparency, self-adhesion and rapid self-healing properties for human motion detection.

Flexible strain sensors have been extensively studied for their potential value in monitoring human activity and health. However, it is still challenging to develop multifunctional flexible strain sensors with simultaneously high transparency, strong self-adhesion, fast self-healing and excellent tensile properties. In this study, we used N,N-dimethylacrylamide (DMA) in the imidazolium-based ionic liquid 1-butyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl] imide ([BMIM][Tf2N]) for "one-step" UV irradiation. A poly(N,N-dimethyl)acrylamide (PDMA) ion-conductive gel was prepared by site polymerization. Based on the good compatibility between PDMA and ionic liquid, the prepared ion-conductive gel has good transparency (∼90%), excellent stretchability (1080%), strong self-adhesion (67.57 kPa), fast self-healing (2 s at room temperature) and great antibacterial activity (∼99% bacterial killing efficiency). Moreover, the strain sensor based on the PDMA ion-conductive gel has good electromechanical performance and can detect different human motions. Based on the simple and easy-to-operate preparation method and the endowed multifunctionality of the PDMA ion-conductive gel, it has broad application prospects in the field of flexible electronic devices.

The impact of human resources for health on the health outcomes of Chinese people.

Human resources for health (HRH) is a cornerstone in the medical system. This paper combined data envelopment analysis (DEA) with Tobit regression analysis to evaluate the efficiency of health care services in China over the years between 2007 and 2019. Efficiency was first estimated by using DEA with the choice of inputs and outputs being specific to health care services and residents' health status. Malmquist index model was selected for estimating the changes in total factor productivity of provinces and exploring whether their performance had improved over the years. Tobit regression model was then employed in which the efficiency score obtained from the DEA computations used as the dependent variable, and HRH was chosen as the independent variables. The results showed that all kinds of health personnel had a significantly positive impact on the efficiency, and more importantly, pharmacists played a critical role in affecting both the provincial and national efficiency. Therefore, the health sector should pay more attention to optimizing allocation of HRH and focusing on professional training of clinical pharmacists.

[Rutin activates AMPKα to ameliorate liver damage caused by perfluorooctanoic acid in mice].

OBJECTIVE: To study the protective effect and mechanism of rutin on perfluorooctanoic acid(PFOA)-induced liver damage in mice. METHODS: A total of 24 male ICR mice were randomly divided into 3 groups, 8 in each group: control group(ctrl group: ddH_2O), perfluorooctanoic acid group(PFOA group: PFOA 20 mg/(kg·d)), rutin intervention group(rutin+PFOA group: PFOA 20 mg/(kg·d)+rutin 20 mg/(kg·d)), normal diet, oral gavage, daily observation, weighting, and recording. After 14 days of treatment, the liver was quickly stripped and weighted after blood sampling and execution. Part of the liver tissue is fixed in 4% paraformaldehyde for HE staining and PAS staining; glutamic-pyruvic transaminase(GPT) activity in the sera samples and triglyceride(TG), total cholesterol(TC), malondialhyde(MDA) content in hepatic tissue homogenate, as well as the activity of some enzymes were assayed; Using western blot to detect adenylate activated protein kinase α(AMPKα) and phosphorylated adenylate-activated protein kinase α(p-AMPKα) expression levels. RESULTS: PFOA caused a significant decrease in the weight of mice, a significant increase in liver weight and liver relative body weight coefficient(P& lt; 0.05), hepatocyte cord dissociation, hepatocyte swelling, dissolution, and obvious damage, and PAS staining positive result were significantly reduced. The GPT activity of the PFOA group was 204.63±11.26 U/L, which was significantly higher than that of the control group(28.80±4.51 U/L)(P& lt; 0.05); The contents of TG, TC and MDA in the liver tissues of mice were 4.89±0.51 mmol/g prot, 8.06±0.84 mmol/g prot and 315.38±60.79 nmol/mg prot, respectively, which were significantly higher than those of the control group(P& lt; 0.05); The activity of T-SOD was 4175.56±334.96 U/mg prot, which was significantly lower than that of the control group(P& lt; 0.05); After rutin intervention, compared with PFOA exposure group, the weight of the mice did not change significantly(P& gt; 0.05), however, the liver weight and the relative weight coefficient of the liver were significantly reduced(P& lt; 0.05), and the hepatocytes swollen and cell lysis were attenuated, the structure of cell cord changed clear, the positive PAS staining cells were significantly increased, the GPT activity of the rutin intervention group was 168.75±18.32 U/L, which was significantly lower than that of the PFOA group(P& lt; 0.05); The contents of TC and MDA in the liver tissues of mice were 4.25±0.77 mmol/g prot and 211.27±44.44 nmol/mg prot, respectively, which were significantly lower than those in the PFOA group(P& lt; 0.05); The T-SOD activity was 7368.88±673.09 U/mg prot, which was significantly higher than that of the PFOA group(P& lt; 0.05). There was no significant difference in AMPKα protein expression in the liver tissues of the three groups. Compared with the PFOA group, the p-AMPKα protein expression in the PFOA+rutin group was significantly up-regulated. CONCLUSION: PFOA exposure can cause liver damage and lipid accumulation in mice. Rutin has a certain protective effect on this damage, which may be related to rutin& apos; s activation of AMPKα protein expression to effectively correct TG levels and enhance the activity of antioxidant enzymes.

[Effect of lentivirus-mediated shRNA targeting down-regulation of PPARα on perfluorododecanoic acid induced oxidative damage in rat hepatocytes BRL 3A].

OBJECTIVE: To investigate the role of peroxisome proliferator-activated receptor alpha(PPARα) in perfluorododecanoic acid(PFDoA)-induced liver oxidative damage in rats by observing lentivirus-mediated shRNA targeting and down-regulating PPARα expression in rat hepatocytes BRL 3 A. METHODS: A PPARα lentivirus-compatible shRNA interference vector Lenti-iPα and a negative control vector Lenti-NC were constructed, and co-transfected with lentivirus packaging helper plasmids into 293 FT cells for lentivirus packaging. The lentivirus stock solution was collected, concentrated and the virus titer was determined. The experimental grouping was as follows, NC-group(infected with negative control lentivirus, without PFDoA exposure), NC+ group(infected with negative control lentivirus, 75 µmol/L PFDoA exposure), iPα-group(infected with interference lentivirus, without PFDoA exposure), iPα+ group(infected with interference lentivirus, 75 µmol/L PFDoA exposure). Rat hepatocytes BRL 3 A cells were treated with lentivirus for 96 h, and then exposed with 75 µmol/L PFDoA in the NC+ group and iPα+ group in the last 24 h. The interference of PPARα in BRL 3 A cells and the role of PPARα in reactive oxygen species(ROS) changes caused by PFDoA were observed. RESULTS: Lentivirus-mediated shRNA successfully achieved targeted downregulation of PPARα expression in BRL 3 A cells. Compared with the NC-group, the mean fluorescence intensity of ROS in rat hepatocytes BRL 3 A in the iPα-group was 12043. 42±808. 58, significantly increased(P<0. 05); The transcription levels of acyl-CoA thioesterases(Acot) 1 gene and its protein expression levels were 0. 43±0. 04 and 0. 34±0. 08, respectively, both significantly decreased(P<0. 05). After PFDoA treatment, compared with NC+ group, the mean fluorescence intensity of ROS in iPα+ group was 12386. 25±356. 36, which also increased significantly(P<0. 05). The transcription levels of Acot1 gene and its protein expression levels were 0. 85±0. 10 and 0. 33±0. 04, respectively, which also decreased significantly(P<0. 05). CONCLUSION: PPARα and its downstream target protein Acot1 may play a role in scavenging ROS in rat hepatocytes BRL 3 A, keeping hepatocytes from oxidative damage caused by foreign substances to the liver.

TRPV1 translocated to astrocytic membrane to promote migration and inflammatory infiltration thus promotes epilepsy after hypoxic ischemia in immature brain.

BACKGROUND: Neonatal hypoxic-ischemic brain damage (HIBD), a leading cause of neonatal mortality, has intractable sequela such as epilepsy that seriously affected the life quality of HIBD survivors. We have previously shown that ion channel dysfunction in the central nervous system played an important role in the process of HIBD-induced epilepsy. Therefore, we continued to validate the underlying mechanisms of TRPV1 as a potential target for epilepsy. METHODS: Neonatal hypoxic ischemia and oxygen-glucose deprivation (OGD) were used to simulate HIBD in vivo and in vitro. Primarily cultured astrocytes were used to assess the expression of TRPV1, glial fibrillary acidic protein (GFAP), cytoskeletal rearrangement, and inflammatory cytokines by using Western blot, q-PCR, and immunofluorescence. Furthermore, brain electrical activity in freely moving mice was recorded by electroencephalography (EEG). TRPV1 current and neuronal excitability were detected by whole-cell patch clamp. RESULTS: Astrocytic TRPV1 translocated to the membrane after OGD. Mechanistically, astrocytic TRPV1 activation increased the inflow of Ca2+, which promoted G-actin polymerized to F-actin, thus promoted astrocyte migration after OGD. Moreover, astrocytic TRPV1 deficiency decreased the production and release of pro-inflammatory cytokines (TNF, IL-6, IL-1ß, and iNOS) after OGD. It could also dramatically attenuate neuronal excitability after OGD and brain electrical activity in HIBD mice. Behavioral testing for seizures after HIBD revealed that TRPV1 knockout mice demonstrated prolonged onset latency, shortened duration, and decreased seizure severity when compared with wild-type mice. CONCLUSIONS: Collectively, TRPV1 promoted astrocyte migration thus helped the infiltration of pro-inflammatory cytokines (TNF, IL-1ß, IL-6, and iNOS) from astrocytes into the vicinity of neurons to promote epilepsy. Our study provides a strong rationale for astrocytic TRPV1 to be a therapeutic target for anti-epileptogenesis after HIBD.

Characterization and Relaxation Properties of a Series of Monodispersed Magnetic Nanoparticles.

Magnetic iron oxide nanoparticles are relatively advanced nanomaterials, and are widely used in biology, physics and medicine, especially as contrast agents for magnetic resonance imaging. Characterization of the properties of magnetic nanoparticles plays an important role in the application of magnetic particles. As a contrast agent, the relaxation rate directly affects image enhancement. We characterized a series of monodispersed magnetic nanoparticles using different methods and measured their relaxation rates using a 0.47 T low-field Nuclear Magnetic Resonance instrument. Generally speaking, the properties of magnetic nanoparticles are closely related to their particle sizes; however, neither longitudinal relaxation rate r 1 nor transverse relaxation rate r 2 changes monotonously with the particle size d . Therefore, size can affect the magnetism of magnetic nanoparticles, but it is not the only factor. Then, we defined the relaxation rates r i ' (i = 1 or 2) using the induced magnetization of magnetic nanoparticles, and found that the correlation relationship between r 1 ' relaxation rate and r 1 relaxation rate is slightly worse, with a correlation coefficient of R 2 = 0.8939, while the correlation relationship between r 2 ' relaxation rate and r 2 relaxation rate is very obvious, with a correlation coefficient of R 2 = 0.9983. The main reason is that r 2 relaxation rate is related to the magnetic field inhomogeneity, produced by magnetic nanoparticles; however r 1 relaxation rate is mainly a result of the direct interaction of hydrogen nucleus in water molecules and the metal ions in magnetic nanoparticles to shorten the T 1 relaxation time, so it is not directly related to magnetic field inhomogeneity.

High expression of Rad51c predicts poor prognostic outcome and induces cell resistance to cisplatin and radiation in non-small cell lung cancer.

Rad51c is critical for homologous recombination repair and genomic stability and may play roles in tumorigenesis and cancer therapy. We investigated the expression level and clinical significance of Rad51c in non-small cell lung cancer (NSCLC) and determined the effect of Rad51c on NSCLC cell chemosensitivity and radiosensitivity. Rad51c expression was detected using immunohistochemistry and was higher in NSCLC patient samples than in adjacent normal tissues. Kaplan-Meier analysis revealed that high Rad51c expression was an independent predictor of short overall survival (OS) and disease-free survival (DFS) in NSCLC patients receiving chemotherapy and/or radiotherapy. Furthermore, Rad51c knockdown increased the killing effect of ionizing radiation (IR) and enhanced cisplatin-induced apoptotic cells in NSCLC cells by disrupting the repair of cisplatin- and IR-induced DNA damage. In addition, ectopic expression of Rad51c dramatically enhanced NSCLC cell resistance to cisplatin and radiotherapy. These findings suggest that increased expression of Rad51c may confer resistance to chemotherapy and/or radiotherapy of NSCLC, and also be an independent prognostic factor for patient outcome. Therefore, targeting Rad51c may represent an improved therapeutic strategy for NSCLC patients with locally advanced disease.

A Digital Lock-In Amplifier for Use at Temperatures of up to 200 °C.

Weak voltage signals cannot be reliably measured using currently available logging tools when these tools are subject to high-temperature (up to 200 °C) environments for prolonged periods. In this paper, we present a digital lock-in amplifier (DLIA) capable of operating at temperatures of up to 200 °C. The DLIA contains a low-noise instrument amplifier and signal acquisition and the corresponding signal processing electronics. The high-temperature stability of the DLIA is achieved by designing system-in-package (SiP) and multi-chip module (MCM) components with low thermal resistances. An effective look-up-table (LUT) method was developed for the lock-in amplifier algorithm, to decrease the complexity of the calculations and generate less heat than the traditional way. The performance of the design was tested by determining the linearity, gain, Q value, and frequency characteristic of the DLIA between 25 and 200 °C. The maximal nonlinear error in the linearity of the DLIA working at 200 °C was about 1.736% when the equivalent input was a sine wave signal with an amplitude of between 94.8 and 1896.0 nV and a frequency of 800 kHz. The tests showed that the DLIA proposed could work effectively in high-temperature environments up to 200 °C.

XRCC3 is a promising target to improve the radiotherapy effect of esophageal squamous cell carcinoma.

Radiotherapy is widely applied for treatment of esophageal squamous cell carcinoma (ESCC). The Rad51-related protein XRCC3 plays roles in the recombinational repair of DNA double-strand breaks to maintain chromosome stability and repair DNA damage. The present study aimed to investigate the effect of XRCC3 on the radiotherapy response of ESCC and the underlying mechanisms of the roles of XRCC3 in ESCC radiosensitivity. XRCC3 expression in ESCC cells and tissues was higher than that in normal esophageal epithelial cells and corresponding adjacent noncancerous esophageal tissue. High XRCC3 expression was positively correlated with resistance to chemoradiotherapy in ESCC and an independent predictor for short disease-specific survival of ESCC patients. Furthermore, the therapeutic efficacy of radiotherapy in vitro and in vivo was substantially increased by knockdown of XRCC3 in ESCC cells. Ectopic overexpression of XRCC3 in both XRCC3-silenced ESCC cells dramatically enhanced ESCC cells' resistance to radiotherapy. Moreover, radiation resistance conferred by XRCC3 was attributed to enhancement of homologous recombination, maintenance of telomere stability, and a reduction of ESCC cell death by radiation-induced apoptosis and mitotic catastrophe. Our data suggest that XRCC3 protects ESCC cells from ionizing radiation-induced death by promoting DNA damage repair and/or enhancing telomere stability. XRCC3 may be a novel radiosensitivity predictor and promising therapeutic target for ESCC.

Endoplasmic reticulum stress involved in high-fat diet and palmitic acid-induced vascular damages and fenofibrate intervention.

Fenofibrate (FF) is widely used to lower blood lipids in clinical practice, but whether its protective effect on endothelium-dependent vasodilatation (EDV) in thoracic aorta is related with endoplasmic reticulum (ER) stress remains unknown. In this study, female Sprauge Dawley rats were divided into standard chow diets (SCD), high-fat diets (HFD) and HFD plus FF treatment group (HFD + FF) randomly. The rats of latter two groups were given HFD feeding for 5 months, then HFD + FF rats were treated with FF (30 mg/kg, once daily) via gavage for another 2 months. The pathological and tensional changes, protein expression of eNOS, and ER stress related genes in thoracic aorta were measured. Then impacts of palmitic acid (PA) and FF on EDV of thoracic aorta from normal female SD rats were observed. Ultimately the expression of ER stress related genes were assessed in primary mouse aortic endothelial cells (MAEC) treated by fenofibric acid (FA) and PA. We found that FF treatment improved serum lipid levels and pathological changes in thoracic aorta, accompanied with decreased ER stress and increased phosphorylation of eNOS. FF pretreatment also improved EDV impaired by different concentrations of PA treatment. The dose- and time-dependent inhibition of cell proliferation by PA were inverted by FA pretreatment. Phosphorylation of eNOS and expression of ER stress related genes were all inverted by FA pretreatment in PA-treated MAEC. Our findings show that fenofibrate recovers damaged EDV by chronic HFD feeding and acute stimulation of PA, this effect is related with decreased ER stress and increased phosphorylation of eNOS.

Fenofibrate treatment attenuated chronic endoplasmic reticulum stress in the liver of nonalcoholic fatty liver disease mice.

Fenofibrate is widely used in clinical practice, but its influence on chronic endoplasmic reticulum (ER) stress induced by feeding a high-calorie and high-cholesterol diet (HCD) has still not been studied. We thus investigated its effects on the liver of the nonalcoholic fatty liver disease (NAFLD) mouse model. Male C57BL/6 mice fed an HCD for 3 months were treated with fenofibrate (HCD + FF, 40 mg/kg, once daily) via gavage for 4 weeks. Insulin sensitivity, serum lipid and inflammatory cytokines were measured. Liver tissues were procured for histological examination as well as analysis of hepatic triglyceride levels, distribution of inflammatory cytokines and genes involved in ER stress. Our results showed that chronic feeding of an HCD successfully induced an NAFLD model accompanied by inflammatory activation, apoptosis and severe ER stress in the liver. Fenofibrate administration significantly improved symptoms of NAFLD and decreased apoptosis, expression of inflammatory cytokines and genes involved in ER stress, such as inositol-requiring enzyme 1α (IRE1α), X-box binding protein 1 (XBP1) and JNK phosphorylation. Thus, our study suggests that fenofibrate protected against inflammatory injury and apoptosis, maybe alleviating ER stress through the IRE1α-XBP1-JNK pathway in the liver of NAFLD mice.

Inhibition of long non-coding RNA NEAT1 impairs myeloid differentiation in acute promyelocytic leukemia cells.

BACKGROUND: Acute promyelocytic leukemia (APL) is characterized by the reciprocal translocation t(15;17), which fuses PML with retinoic acid receptor alpha (RARα). Although PML-RARα is crucially important for pathogenesis and responsiveness to treatment, the molecular and cellular mechanisms by which PML-RARα exerts its oncogenic potential have not been fully elucidated. Recent reports have suggested that long non-coding RNAs (lncRNAs) contribute to the precise control of gene expression and are involved in human diseases. Little is known about the role of lncRNA in APL. METHODS: We analyzed NEAT1 expression in APL samples and cell lines by real-time quantitative reverse transcription-PCR (qRT-PCR). The expression of PML-RARα was measured by Western blot. Cell differentiation was assessed by measuring the surface CD11b antigen expression by flow cytometry analysis. RESULTS: We found that nuclear enriched abundant transcript 1 (NEAT1), a lncRNA essential for the formation of nuclear body paraspeckles, is significantly repressed in de novo APL samples compared with those of healthy donors. We further provide evidence that NEAT1 expression was repressed by PML-RARα. Furthermore, significant NEAT1 upregulation was observed during all-trans retinoic acid (ATRA)-induced NB4 cell differentiation. Finally, we demonstrate the importance of NEAT1 in myeloid differentiation. We show that reduction of NEAT1 by small interfering RNA (siRNA) blocks ATRA-induced differentiation. CONCLUSIONS: Our results indicate that reduced expression of the nuclear long noncoding RNA NEAT1 may play a role in the myeloid differentiation of APL cells.

Network pharmacology and molecular docking-based strategy for predicting anti-tumour mechanism of linarin.

The aim was to explore the anti-tumour mechanism of linarin (LIN) based on network pharmacology and molecular docking. PharmMapper database and GeneCards database were used to screen anti-tumour related targets of LIN. Enrichment analysis of GO and KEGG was conducted to predict the key targets and pathways. At last, LIN was docked with the key targets. ESR1, ESR2, EGFR, AR, TGFBR2, F2, MAPK10, MAPK14, CDK2 and HSP90AA1 were identified as the key targets. The key pathways included pathways in cancer, prostate cancer, pancreatic cancer and breast cancer. KEGG pathway maps indicated that the anti-tumour effect of LIN may be mainly achieved by intervening related targets in the following pathways: AR-HSP/AR-AR/PSA/proliferation and evading apoptosis;F2/GPCR/…/ROCK/tissue invasion and metastasis;F2/GPCR/…/Raf/MAPK signalling pathway/proliferation and sustained angiogenesis; EGFR/Grb2/…/Raf/MAPK signalling pathway/proliferation and sustained angiogenesis; ER/Oestrogen signalling pathway/proliferation;TGFBR2/Smad2/3/TGF-ß signalling pathway/insensitivity to anti-growth signals; oxidative stress/KEAP1/NRF2/…/proliferation and evading apoptosis. LIN had strong binding activity with ESR2, EGFR, AR, CDK2 and HSP90AA1.

Fabrication, characterization, and application of pea protein-based edible film enhanced by oregano essential oil (OEO) micro- or nano-emulsion.

Pea protein isolate (PPI)-based active films were prepared by incorporating 0.5 %, 1.0 %, or 2.0 % of oregano essential oil (OEO), either in the form of micro-emulsion (MOEO) or nano-emulsion (NOEO). The particle size and polydispersity index of OEO droplets were 2755.00 nm and 0.63 for MOEO, and 256.30 nm and 0.20 for NOEO. The surface and cross-sectional SEM results revealed the presence of holes and internal pores within the film upon the addition of OEO. The molecular interaction between PPI and OEO was confirmed by FTIR. The addition of OEO significantly increased film thickness, decreased water contact angle, and imparted a more yellow color. At a low concentration (0.5 %), the addition of OEO significantly improved the water vapor barrier and mechanical properties of the film. However, at higher concentrations, these film properties were significantly weakened. Additionally, the film antimicrobial properties were assessed after OEO addition. In vitro inhibition zone results indicated that a 2.0 % addition of OEO significantly suppressed the growth of three Salmonella strains [Salmonella Typhimurium (ATCC14028), Salmonella Infantis 94-1, and Salmonella Enteritidis PT-30]. Application of pea protein-based film with 2.0 % OEO on chicken breast demonstrated significant reduction in microbial count. Our results further showed that reducing the particle size of OEO from micrometer-scale to nanometer-scale in the PPI film matrix did not significantly alter film properties or antimicrobial activities. The study demonstrated that the antibacterial film based on pea protein and OEO is an innovative food packing material for prohibiting bacteria growth on poultry products.