Identification and characterization of extrachromosomal circular DNA in large-artery atherosclerotic stroke.

Extrachromosomal circular DNA (eccDNA) is a new biomarker and regulator of diseases. However, the role of eccDNAs in large-artery atherosclerotic (LAA) stroke remains unclear. Through high-throughput circle-sequencing technique, the length distribution, genomic characteristic and motifs feature of plasma eccDNA from healthy controls (CON) and patients with LAA stroke were analysed. Then, the potential functions of the annotated eccDNAs were investigated using GO and KEGG pathway analyses. EccDNAs mapped to the reference genome showed SHN3 and BCL6 were LAA stroke unique transcription factors. The genes of differentially expressed eccDNAs between LAA stroke patients and CON were mainly involved in axon/dendrite/neuron projection development and maintenance of cellular structure via Wnt, Rap1 and MAPK pathways. Moreover, LAA stroke unique eccDNA genes played a role in regulation of coagulation and fibrinolysis, and there were five LAA stroke unique eccDNAs (Chr2:12724406-12724784, Chr4:1867120-186272046, Chr4:186271494-186271696, Chr7:116560296-116560685 and Chr11:57611780-5761192). Additionally, POLR2C and AURKA carried by ecDNAs (eccDNA size >100 kb) of LAA stroke patients were significantly associated with development of LAA stroke. Our data firstly revealed the characteristics of eccDNA in LAA stroke and the functions of LAA stroke unique eccDNAs and eccDNA genes, suggesting eccDNA is a novel biomarker and mechanism of LAA stroke.

Signature and function of plasma exosome-derived circular RNAs in patients with hypertensive intracerebral hemorrhage.

Intracerebral hemorrhage (ICH) is one of the major causes of death and disability, and hypertensive ICH (HICH) is the most common type of ICH. Currently, the outcomes of HICH patients remain poor after treatment, and early prognosis prediction of HICH is important. However, there are limited effective clinical treatments and biomarkers for HICH patients. Although circRNA has been widely studied in diseases, the role of plasma exosomal circRNAs in HICH remains unknown. The present study was conducted to investigate the characteristics and function of plasma exosomal circRNAs in six HICH patients using circRNA microarray and bioinformatics analysis. The results showed that there were 499 differentially expressed exosomal circRNAs between the HICH patients and control subjects. According to GO annotation and KEGG pathway analyses, the targets regulated by differentially expressed exosomal circRNAs were tightly related to the development of HICH via nerve/neuronal growth, neuroinflammation and endothelial homeostasis. And the differentially expressed exosomal circRNAs could mainly bind to four RNA-binding proteins (EIF4A3, FMRP, AGO2 and HUR). Moreover, of differentially expressed exosomal circRNAs, hsa_circ_00054843, hsa_circ_0010493 and hsa_circ_00090516 were significantly associated with bleeding volume and Glasgow Coma Scale score of the subjects. Our findings firstly revealed that the plasma exosomal circRNAs are significantly involved in the progression of HICH, and could be potent biomarkers for HICH. This provides the basis for further research to pinpoint the best biomarkers and illustrate the mechanism of exosomal circRNAs in HICH.

CircRNA and Stroke: New Insight of Potential Biomarkers and Therapeutic Targets.

Stroke, the second-largest cause of death and the leading cause of disability globally, presents significant challenges in terms of prognosis and treatment. Identifying reliable prognosis biomarkers and treatment targets is crucial to address these challenges. Circular RNA (circRNA) has emerged as a promising research biomarkers and therapeutic targets because of its tissue specificity and conservation. However, the potential role of circRNA in stroke prognosis and treatment remains largely unexplored. This review briefly elucidate the mechanism underlying circRNA's involvement in stroke pathophysiology. Additionally, this review summarizes the impact of circRNA on different forms of strokes, including ischemic stroke and hemorrhagic stroke. And, this article discusses the positive effects of circRNA on promoting cerebrovascular repair and regeneration, maintaining the integrity of the blood-brain barrier (BBB), and reducing neuronal injury and immune inflammatory response. In conclusion, the significance of circRNA as a potential prognostic biomarker and a viable therapeutic target was underscored.

Thromboelastography-Guided Antiplatelet Therapy for Patients with Ischemic Cerebrocardiovascular Diseases: A Systematic Review and Meta-Analysis.

INTRODUCTION: The effectiveness of thromboelastography (TEG)-guided antiplatelet therapy in patients with ischemic cerebrocardiovascular diseases is not well-established. This systematic review evaluates the efficacy and safety of TEG-guided antiplatelet therapy compared to standard treatment in patients with ischemic cerebrocardiovascular diseases. METHODS: Randomized controlled trials (RCTs) and observational studies comparing TEG-guided antiplatelet therapy with standard therapy in patients suffering from ischemic stroke (IS) or coronary artery disease (CAD) were identified. The primary efficacy measure was a composite of ischemic and hemorrhagic events. Secondary efficacy measures included any ischemic events, while safety was assessed by the occurrence of bleeding events. RESULTS: Ten studies involving 4 RCTs and 6 observational studies with a total of 1,678 patients were included. When considering a composite of ischemic and hemorrhagic events in RCTs, a significant reduction was observed in IS or CAD patients under TEG-guided therapy compared to standard therapy (OR: 0.45, 95% CI: 0.27-0.75, p = 0.002). After pooling RCTs and observational studies together, compared to standard antiplatelet therapy, TEG-guided therapy significantly reduced the risk of a composite of ischemic and hemorrhagic events (OR: 0.26, 95% CI: 0.19-0.37; p < 0.00001), ischemic events (OR: 0.28, 95% CI: 0.19-0.41; p < 0.00001), and bleeding events (OR: 0.31, 95% CI: 0.16-0.62; p = 0.0009) in patients with IS or CAD. CONCLUSION: TEG-guided antiplatelet therapy appears to be both effective and safe for patients with IS or CAD. These findings support the use of TEG testing to tailor antiplatelet therapy in individuals with ischemic cerebrocardiovascular diseases.

Lower Plasma miR-223 Level Is Associated with Clopidogrel Resistance in Acute Coronary Syndrome: A Systematic Review and Meta-Analysis.

Objectives: To evaluate the relationship between the plasma miR-223 expression level and clopidogrel resistance in acute coronary syndrome (ACS) patients. Methods: We performed a search for publications using online databases including PubMed, EMBASE, Cochrane Library, and Chinese Databases (CNKI database, Weipu database, and Wanfang database) from the inception of the databases to June 18, 2023, to identify studies reporting the relationship between the plasma miR-223 level and clopidogrel resistance in ACS patients. Two researchers independently searched and screened to ensure the consistency of the results and assess the quality of the included studies according to the Newcastle-Ottawa scale. A fixed-effects model was used for pooling data with STATA 14.0. Results: Four articles including 399 Chinese ACS patients were eligible for the meta-analysis. Low plasma miR-223 levels were independently correlated with clopidogrel resistance in Chinese ACS patients (OR 0.58, 95% CI: 0.33-1.04). Conclusion: Lower plasma miR-223 levels are associated with clopidogrel resistance in Chinese ACS patients, suggesting that miR-223 may be a potential diagnostic biomarker of clopidogrel resistance.

Flavokavain C Suppresses Breast Cancer Cell Viability and Induces Cell Apoptosis by Triggering DNA Damage.

Breast cancer, presented by multiple breast cancer subtypes that coexist within a diagnosed tumor in clinical, has ranked as the most common malignancy in women in recent years. Evidence suggested that limited effective drugs caused the unsatisfactory therapeutic efficacy of breast cancer. Flavokavain C exhibited anticancer activity on colon cancer cells HCT116. It is yet unknown if it can be used to treat breast cancer. This study aims to believe the mechanisms by which Flavokavain C suppresses cell proliferation and the pathways that impact on this effect in breast cancer. 3-(4,5-Dimethythiazol)-2,5-diphenyltetrazolium bromide assay was chosen to evaluate cell cytotoxicity. Colony formation and cell proliferation assays using 5-ethynyl-2'-deoxyuridine staining were performed. Cell cycle progression and apoptosis were examined via flow cytometry and Western blotting, respectively. Five methods (comet assay, immunofluorescence, Western blotting, agarose gel electrophoresis and molecular docking) were used to quantify DNA damage and its cellular response. Compared to cisplatin, Flavokavain C possessed a comparable or more substantial inhibitory effect on breast cancer cell viability while having lower cytotoxicity on human mammary cells. Breast cancer cells treated with Flavokavain C had their colony formation suppressed, DNA replication blocked, the G2/M phase cell cycle arrested, and apoptosis. Furthermore, the results indicated that Flavokavain C would directly interact with DNA and induce DNA cleavage, demonstrating that DNA is an attractive substrate for Flavokavain C. These results suggested that Flavokavain C had strong anticancer activity against multiple subtypes of breast cancer cells.

A stochastic agent-based model to evaluate COVID-19 transmission influenced by human mobility.

The COVID-19 pandemic has created an urgent need for mathematical models that can project epidemic trends and evaluate the effectiveness of mitigation strategies. A major challenge in forecasting the transmission of COVID-19 is the accurate assessment of the multiscale human mobility and how it impacts infection through close contacts. By combining the stochastic agent-based modeling strategy and hierarchical structures of spatial containers corresponding to the notion of geographical places, this study proposes a novel model, Mob-Cov, to study the impact of human traveling behavior and individual health conditions on the disease outbreak and the probability of zero-COVID in the population. Specifically, individuals perform power law-type local movements within a container and global transport between different-level containers. It is revealed that frequent long-distance movements inside a small-level container (e.g., a road or a county) and a small population size reduce both the local crowdedness and disease transmission. It takes only half of the time to induce global disease outbreaks when the population increases from 150 to 500 (normalized unit). When the exponent c1 of the long-tail distribution of distance k moved in the same-level container, p(k)∼k-c1·level, increases, the outbreak time decreases rapidly from 75 to 25 (normalized unit). In contrast, travel between large-level containers (e.g., cities and nations) facilitates global spread of the disease and outbreak. When the mean traveling distance across containers 1d increases from 0.5 to 1 (normalized unit), the outbreak occurs almost twice as fast. Moreover, dynamic infection and recovery in the population are able to drive the bifurcation of the system to a "zero-COVID" state or to a "live with COVID" state, depending on the mobility patterns, population number and health conditions. Reducing population size and restricting global travel help achieve zero-COVID-19. Specifically, when c1 is smaller than 0.2, the ratio of people with low levels of mobility is larger than 80% and the population size is smaller than 400, zero-COVID can be achieved within fewer than 1000 time steps. In summary, the Mob-Cov model considers more realistic human mobility at a wide range of spatial scales, and has been designed with equal emphasis on performance, low simulation cost, accuracy, ease of use and flexibility. It is a useful tool for researchers and politicians to apply when investigating pandemic dynamics and when planning actions against disease. Supplementary Information: The online version contains supplementary material available at 10.1007/s11071-023-08489-5.

Analysis of the miRNA expression profile of laboratory red crucian carp under low-dose caesium-137 irradiation.

Radiation can cause the differential expression of biological miRNA molecules. This research was based on the development of the laboratory red crucian carp (LRCC) to explore the feasibility of its application in the detection of low-dose ionizing radiation-induced biological damage in aquatic environments and the development of related molecular markers. Adult LRCC were irradiated with caesium-137 at 0.3 Gy, while RNA-seq and bioinformatics techniques were used to identify miRNAs that were differentially expressed relative to their levels in the nonirradiation group. Analysis of liver sections showed that liver cells in the radiation group showed nuclear pyknosis. In this study, 34 miRNAs differentially expressed in the liver of LRCC after irradiation were identified, among which seven were new crucian carp miRNAs; a total of 632 target genes were predicted in the prediction analysis. The results of comprehensive GO enrichment and KEGG pathway analyses showed that these target genes were mainly involved in energy transfer and material catabolism, especially malonyl-CoA biosynthesis, acetyl-CoA carboxylase activity, fatty acid biosynthesis and metabolism, and pyruvate metabolism; in addition, the AMPK signalling pathway was the most active pathway. This study shows that the LRCC is sensitive to radiation, or can be used as a candidate experimental animal to study the biological effects of radiation, and the screened miRNA can be used as a pre-selected biomarker for radiation damage detection and radiation biological environmental monitoring. CLINICAL TRIALS REGISTRATION: None.

A microfluidic generator of dynamic shear stress and biochemical signals based on autonomously oscillatory flow.

Biological cells in vivo typically reside in a dynamic flowing microenvironment with extensive biomechanical and biochemical cues varying in time and space. These dynamic biomechanical and biochemical signals together act to regulate cellular behaviors and functions. Microfluidic technology is an important experimental platform for mimicking extracellular flowing microenvironment in vitro. However, most existing microfluidic chips for generating dynamic shear stress and biochemical signals require expensive, large peripheral pumps and external control systems, unsuitable for being placed inside cell incubators to conduct cell biology experiments. This study has developed a microfluidic generator of dynamic shear stress and biochemical signals based on autonomously oscillatory flow. Further, based on the lumped-parameter and distributed-parameter models of multiscale fluid dynamics, the oscillatory flow field and the concentration field of biochemical factors has been simulated at the cell culture region within the designed microfluidic chip. Using the constructed experimental system, the feasibility of the designed microfluidic chip has been validated by simulating biochemical factors with red dye. The simulation results demonstrate that dynamic shear stress and biochemical signals with adjustable period and amplitude can be generated at the cell culture chamber within the microfluidic chip. The amplitudes of dynamic shear stress and biochemical signals is proportional to the pressure difference and inversely proportional to the flow resistance, while their periods are correlated positively with the flow capacity and the flow resistance. The experimental results reveal the feasibility of the designed microfluidic chip. Conclusively, the proposed microfluidic generator based on autonomously oscillatory flow can generate dynamic shear stress and biochemical signals without peripheral pumps and external control systems. In addition to reducing the experimental cost, due to the tiny volume, it is beneficial to be integrated into cell incubators for cell biology experiments. Thus, the proposed microfluidic chip provides a novel experimental platform for cell biology investigations.

A microfluidic system for precisely reproducing physiological blood pressure and wall shear stress to endothelial cells.

To reproduce hemodynamic stress microenvironments of endothelial cells in vitro is of vital significance, by which one could exploit the quantitative impact of hemodynamic stresses on endothelial function and seek innovative approaches to prevent circulatory system diseases. Although microfluidic technology has been regarded as an effective method to create physiological microenvironments, a microfluidic system to precisely reproduce physiological arterial hemodynamic stress microenvironments has not been reported yet. In this paper, a novel microfluidic chip consisting of a cell culture chamber with on-chip afterload components designed by the principle of input impedance to mimic the global hemodynamic behaviors is proposed. An external feedback control system is developed to accurately generate the input pressure waveform. A lumped parameter hemodynamic model (LPHM) is built to represent the input impedance to mimic the on-chip global hemodynamic behaviors. Sensitivity analysis of the model parameters is also elaborated. The performance of reproducing physiological blood pressure and wall shear stress is validated by both numerical characterization and flow experiment. Investigation of intracellular calcium ion dynamics in human umbilical vein endothelial cells is finally conducted to demonstrate the biological applicability of the proposed microfluidic system.

Attenuated Cardiac oxidative stress, inflammation and apoptosis in Obese Mice with nonfatal infection of Escherichia coli.

Obesity is a risk factor of many diseases, but could be beneficial to the individuals with bacterial infection. The present study was conducted to investigate the relationship between obesity and heart during nonfatal bacterial infection. Male normal (lean) and diet-induced obesity mice (DIO, fed with high-fat diet) were chosen to perform nasal instillation with E. coli to establish a nonfatal acute mouse model. The cardiac histopathology, inflammation and oxidative damage, as well as apoptosis were detected post-infection. The results revealed that the Escherichia coli (E.coli)-infected mice exhibited increased cardiac index, contents of IL-1ß, IL-6, IL-8, TNF-α, leptin and resistin, levels of apoptotic proteins (caspase-3 and caspase-9, and bax/bcl-2 ratio), cardiac pathological changes and oxidative stress. Furthermore, these parameters were more serious in the lean mice than those in the DIO mice. In summary, our findings gave a new sight that E.coli infection impaired heart via histopathological lesions, inflammation and oxidative stress and excessive apoptosis of cardiomyocytes. Interestingly, obesity exerted attenuated effects on the heart of mice with non-fatal infection of E.coli through decreased inflammation, oxidative stress and apoptosis of cardiac tissue.

Vitamin E protects against cadmium-induced sub-chronic liver injury associated with the inhibition of oxidative stress and activation of Nrf2 pathway.

Hepatic oxidative stress, as one important mechanism of cadmium (Cd)-induced hepatic toxicity, could, as known, be ameliorated by vitamin E (VE). However, the underlying mechanism remains to be elucidated. To investigate whether the antioxidant vitamin E can protect against Cd-induced sub-chronic liver injury associated with oxidative stress and nuclear factor erythrocyte 2-related factor 2 (Nrf2) pathway, male Sprague-Dawley rats (nine-week-old) were randomly divided into four groups (eight rats/group), namely, control, VE (100 mg/kg VE), Cd (5 mg/kg CdCl2) and VE+Cd (100 mg/kg VE+5 mg/kg CdCl2), and received intragastric administration of Cd and/or VE for four weeks. Cd-exposure alone resulted in reduced liver weight, liver histological alteration and oxidative stress, accumulation of Cd in the liver, elevated ALT and AST concentrations in serum together with decreased mRNA and protein expressions of Nrf2 pathway related molecules (Nrf2, HO-1, NQO-1, GCLC, GCLM and GST). However, the co-treatment of Cd and VE significantly ameliorated the changes mentioned above, and promoted the expression of genes and proteins of Nrf2 pathway related molecules in comparison to the Cd-exposure alone. Our results indicate that the protective effect of VE against Cd-induced sub-chronic hepatic damage in rats is associated with the inhibition of oxidative stress and activation of Nrf2 pathway.

Puerarin Loaded PLGA Nanoparticles: Optimization Processes of Preparation and Anti-alcohol Intoxication Effects in Mice.

To improve the bioavailability of puerarin in liver, the optimized preparation method of puerarin-PLGA nanoparticles (Pue-PLGA-nps) and the effect of Pue-PLGA-nps on alcoholism mice were studied. The preparation of Pue-PLGA-nps was optimized by the Box-Behnken design and response surface methodology (RSM). To estimate the anti-alcoholism of Pue-PLGA-nps in vivo, drunkenness incubation period and sober time of mice were detected, and Morris water maze (MWM) test was performed. AST, ALT, and SOD were used to determine the damages and oxidative stress in the liver, as well as histopathological observation of the liver. The optimal preparation conditions of Pue-PLGA-nps in RSM were as follows: the drug-material ratio was 1:1.4, the reaction temperature was 65°C, and the reaction time was 13 min. The drug entrapment efficiency of Pue-PLGA-nps was 90.6% and closely up to 98.9% of the standard prediction value. The results in vivo showed that the Pue-PLGA-nps significantly increased the drunkenness incubation period in comparison with the model group and decreased drunkenness sober time and landing time in MWM in comparison with the model group and puerarin group (P<0.05) . The contents of AST and ALT in the liver of Pue-PLGA-nps group were significantly lower than those of model group and Puerarin group (P<0.05), and the activity of SOD in the liver of Pue-PLGA-nps group was higher than that of model group (P<0.05). By histopathological observation, moreover, Pue-PLGA-nps significantly attenuated the impairment of the liver caused by alcoholism. In conclusion, through BBD and RSM, the process conditions of the Pue-PLGA-nps were successfully optimized. The Pue-PLGA-nps exerted higher bioavailability and better effect of anti-alcoholism than puerarin, indicating PLGA nanoparticles could be potential to deliver drug.

Acoustic Separation of Nanoparticles in Continuous Flow.

The separation of nanoscale particles based on their differences in size is an essential technique to the nanoscience and nanotechnology community. Here, nanoparticles are successfully separated in a continuous flow by using tilted-angle standing surface acoustic waves. The acoustic field deflects nanoparticles based on volume, and the fractionation of nanoparticles is optimized by tuning the cutoff parameters. The continuous separation of nanoparticlesis demonstrated with a ≈90% recovery rate. The acoustic nanoparticle separation method is versatile, non-invasive, and simple.

Commensal Bacteria-induced Interleukin 1ß (IL-1ß) Secreted by Macrophages Up-regulates Hepcidin Expression in Hepatocytes by Activating the Bone Morphogenetic Protein Signaling Pathway.

The liver hormone hepcidin is the central regulator of systemic iron metabolism. Its increased expression in inflammatory states leads to hypoferremia and anemia. Elucidation of the mechanisms that up-regulate hepcidin during inflammation is essential for developing rational therapies for this anemia. Using mouse models of inflammatory bowel disease, we have shown previously that colitis-associated hepcidin induction is influenced by intestinal microbiota composition. Here we investigate how two commensal bacteria, Bifidobacterium longum and Bacteroides fragilis, representative members of the gut microbiota, affect hepcidin expression. We found that supernatants of a human macrophage cell line infected with either of the bacteria up-regulated hepcidin when added to a human hepatocyte cell line. This activity was abrogated by neutralization of IL-1ß. Moreover, purified IL-1ß increased hepcidin expression when added to the hepatocyte line or primary human hepatocytes and when injected into mice. IL-1ß activated the bone morphogenetic protein (BMP) signaling pathway in hepatocytes and in mouse liver, as indicated by increased phosphorylation of small mothers against decapentaplegic proteins. Activation of BMP signaling correlated with IL-1ß-induced expression of BMP2 in human hepatocytes and activin B in mouse liver. Treatment of hepatocytes with two different chemical inhibitors of BMP signaling or with a neutralizing antibody to BMP2 prevented IL-1ß-induced up-regulation of hepcidin. Our results clarify how commensal bacteria affect hepcidin expression and reveal a novel connection between IL-1ß and activation of BMP signaling. They also suggest that there may be differences between mice and humans with respect to the mechanism by which IL-1ß up-regulates hepcidin.

Aflatoxin B1 affects apoptosis and expression of Bax, Bcl-2, and Caspase-3 in thymus and bursa of fabricius in broiler chickens.

Aflatoxin B1 is known as a mycotoxin that develops various health problems of animals, the effects of AFB1 on thymus and bursa of Fabricius in chickens are not clear. The objective of this study was to investigate the apoptosis of thymus and bursa of Fabricius in broilers fed with AFB1 . Two hundred Avian broilers were randomly divided into four groups of 50 each, namely control group and three AFB1 groups fed with 0.15 mg, 0.3 mg, and 0.6 mg AFB1 /kg diet, respectively. In this study, flow cytometer and immunohistochemical approaches were used to determine the percentage of apoptotic cells and the expression of Bax, Bcl-2, and Caspase-3. The results showed that consumption of AFB1 diets results in increased percentage of apoptotic cells and increased expression of Caspase-3 in both thymus and bursa of Fabricius. The expression of Bax was increased and the expression of Bcl-2 was decreased in the thymus, but no significant changes in Bax and Bcl-2 expression were observed in the bursa of Fabricius when broilers fed with AFB1 . These findings suggest that adverse effects of AFB1 on thymus and bursa of Fabricius in broilers were confirmed by increased apoptotic cells and abnormal expression of Caspase-3. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1113-1120, 2016.

Effects of aflatoxin B1 on oxidative stress markers and apoptosis of spleens in broilers.

The purpose of the present study was to investigate the oxidative damage and apoptosis induced by aflatoxin B1 (AFB1) in spleen of broilers. A total of 200 one-day-old avian male broilers were randomly divided into 4 equal groups of 50 each and were fed for 21 days as follows: a control diet and three AFB1 diets containing 0.15, 0.3, and 0.6 mg AFB1/kg diet. Consumption of AFB1 diets induced oxidative stress in the spleen of chicken as evidenced by reduced glutathione peroxidase, glutathione reductase, and catalase activities, decreased glutathione contents, and increased malondialdehyde contents in explaining the pathogenesis. Flow cytometer method and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling assay revealed that the apoptotic splenocytes were increased in AFB1 groups. The results suggest that AFB1 induced excessive apoptosis of splenic lymphocytes, which is correlated with increased oxidative stress. The present results may be helpful for explaining the pathogenesis of AFB1-induced immunosuppression.

Modulation of the PI3K/Akt Pathway and Bcl-2 Family Proteins Involved in Chicken's Tubular Apoptosis Induced by Nickel Chloride (NiCl2).

Exposure of people and animals to environments highly polluted with nickel (Ni) can cause pathologic effects. Ni compounds can induce apoptosis, but the mechanism and the pathway of Ni compounds-induced apoptosis are unclear. We evaluated the alterations of apoptosis, mitochondrial membrane potential (MMP), phosphoinositide-3-kinase (PI3K)/serine-threonine kinase (Akt) pathway, and Bcl-2 family proteins induced by nickel chloride (NiCl2) in the kidneys of broiler chickens, using flow cytometry, terminal deoxynucleotidyl transferase 2'-deoxyuridine 5'-triphosphate dUTP nick end-labeling (TUNEL), immunohistochemstry and quantitative real-time polymerase chain reaction (qRT-PCR). We found that dietary NiCl2 in excess of 300 mg/kg resulted in a significant increase in apoptosis, which was associated with decrease in MMP, and increase in apoptosis inducing factor (AIF) and endonuclease G (EndoG) protein and mRNA expression. Concurrently, NiCl2 inhibited the PI3K/Akt pathway, which was characterized by decreasing PI3K, Akt1 and Akt2 mRNA expression levels. NiCl2 also reduced the protein and mRNA expression of anti-apoptotic Bcl-2 and Bcl-xL and increased the protein and mRNA expression of pro-apoptotic Bax and Bak. These results show that NiCl2 causes mitochondrial-mediated apoptosis by disruption of MMP and increased expression of AIF and EndoG mRNA and protein, and that the underlying mechanism of MMP loss involves the Bcl-2 family proteins modulation and PI3K/Akt pathway inhibition.

Cross border e-commerce development and enterprise digital technology innovation-empirical evidence from listed companies in China.

As the main form of digital trade, cross-border e-commerce plays an important role, allowing China to expand its opening-up and promote the optimal foreign trade structure. It also provides opportunities for Chinese enterprises to develop digital technology. From the perspective of the establishment of China's cross-border e-commerce comprehensive pilot zone (CBECPZ), this article uses the multi-period DID method to examine the effects of cross-border e-commerce on enterprise digital technology innovation based on listed companies in the Shanghai and Shenzhen stock markets from 2007 to 2020. The CBECPZ dramatically promotes enterprise digital technology innovation. The mechanism test shows that the CBECPZ promotes digital technology innovation by financing constraint alleviation, digital transformation, and producer service industry agglomeration. The heterogeneity test shows that the direct effect is more significant in the enterprises of large-scale, non-state-owned, with high ICT correlation and in areas with strong government resource allocation capabilities. The research findings have important reference value for how to utilize cross-border e-commerce to promote digital technology innovation, and they also provide directional references for other developing countries to develop cross-border e-commerce.