Inhibition of ROS/caspase-3/GSDME-mediated pyroptosis alleviates high glucose-induced injury in AML-12 cells.

Diabetic liver injury (DLI) is a chronic complication of the liver caused by diabetes, and its has become one of the main causes of nonalcoholic fatty liver disease (NAFLD). The gasdermin E (GSDME)-dependent pyroptosis signaling pathway is involved in various physiological and pathological processes; however, its role and mechanism in DLI are still unknown. This study was performed to investigate the role of GSDME-mediated pyroptosis in AML-12 cell injury induced by high glucose and to evaluate the therapeutic potential of caspase-3 inhibition for DLI. The results showed that high glucose activated apoptosis by regulating the apoptotic protein levels including Bax, Bcl-2, and enhanced cleavage of caspase-3 and PARP. Notably, some of the hepatocytes treated with high glucose became swollen, accompanied by GSDME-N generation, indicating that pyroptosis was further induced by active caspase-3. Moreover, the effects of high glucose on AML-12 cells could be partly reversed by a reactive oxygen scavenger (NAC) and caspase-3 specific inhibitor (Z-DEVD-FMK), which suggests high glucose induced GSDME-dependent pyroptosis in AML-12 cells through increasing ROS levels and activating caspase-3. In conclusion, our results show that high glucose can induce pyroptosis in AML-12 cells, at least in part, through the ROS/caspase-3/GSDME pathway，and inhibition of caspase-3 can ameliorate high glucose-induced hepatocyte injury, providing an important basis for clarifying the pathogenesis and treatment of DLI.

Emerging programmable nuclease-based detection for food safety.

Food safety issues are an important challenge across the world. Programmable nucleases are emerging as new tools because of their significant biological advantages. This forum article provides an overview of recent advances and challenges in the novel paradigm of programmable nuclease-based detection for food safety.

Recent advances of food safety detection by nucleic acid isothermal amplification integrated with CRISPR/Cas.

Food safety problems have become one of the most important public health issues worldwide. Therefore, the development of rapid, effective and robust detection is of great importance. Amongst a range of methods, nucleic acid isothermal amplification (NAIA) plays a great role in food safety detection. However, the widespread application remains limited due to a few shortcomings. CRISPR/Cas system has emerged as a powerful tool in nucleic acid detection, which could be readily integrated with NAIA to improve the detection sensitivity, specificity, adaptability versatility and dependability. However, currently there was a lack of a comprehensive summary regarding the integration of NAIA and CRISPR/Cas in the field of food safety detection. In this review, the recent advances in food safety detection based on CRISPR/Cas-integrated NAIA were comprehensively reviewed. To begin with, the development of NAIA was summarized. Then, the types and working principles of CRISPR/Cas were introduced. The applications of the integration of NAIA and CRISPR/Cas for food safety were mainly introduced and objectively discussed. Lastly, current challenges and future opportunities were proposed. In summary, this technology is expected to become an important approach for food safety detection, leading to a safer and more reliable food industry.

Ghrelin protects against ischemia/reperfusion-induced hepatic injury via inhibiting Caspase-11-mediated noncanonical pyroptosis.

BACKGROUND: Ischemia/reperfusion (I/R) injury is a complication of liver transplantation. I/R-induced inflammatory cell death, namely, pyroptosis, that is triggered by overactive inflammasomes results in the production of proinflammatory cytokines. Hepatic I/R injury correlates with the activation of the Caspase-11-mediated pyroptosis pathway. We investigated whether ghrelin, which is a pleiotropic gut hormone, may have anti-hepatic I/R injury effects, but the mechanism by which Ghrelin ameliorates hepatic I/R -induced injury remains a mystery. METHODS: Hepatic I/R injury was induced in a mouse model by clamping the left and right lobes of the liver for 90 min followed by reperfusion for 6 h, 12 h, or 24 h. As treatment, a saline with or without ghrelin was infused via the tail vain. Hepatocytes were isolated using a two-step collagenase liver perfusion method. RESULTS: In our study, treatment with ghrelin protected against hepatic I/R injury as shown by decreased alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) levels (p < 0.001) and reduced the histological injury in liver tissues compared with untreated controls. The LDH level of primary hepatocytes was increased by hypoxia/reoxygenation (H/R), and it was then restored to normal levels by ghrelin-treatment (p < 0.05). Western blotting analysis showed that ghrelin significantly inhibited the expression of pyroptosis-related proteins, including Caspase-11, GSDMD-N, NLRP3 and HMGB1, both in vivo and in vitro (all p < 0.05) compared with the untreated controls. Immunofluorescence showed that the expression of Gasdamin D (GSDMD) in hepatocytes was increased after I/R or H/R, whereas GSDMD expression was reduced by ghrelin treatment (p < 0.05). CONCLUSIONS: Our findings suggest that ghrelin ameliorated I/R-induced hepatic injury by inhibiting Caspase-11-mediated pyroptosis. Ghrelin may be a potential therapeutic option to prevent hepatic I/R injury after liver transplantation.

Litchi procyanidins inhibit colon cancer proliferation and metastasis by triggering gut-lung axis immunotherapy.

Litchi chinensis seed, as a valuable by-product of the subtropical fruit litchi (Litchi chinensis Sonn.), has been confirmed to be rich in procyanidins (LPC). The anticarcinogenic properties of procyanidins has been primarily attributed to their antioxidant and anti-inflammatory activities. However, there is a comparative paucity of information on if and how LPC inhibits colon cancer. Here, LPC significantly inhibited CT26 colon cancer cells proliferation and metastasis in vivo and in vitro. In CT26 lung metastatic mice, the anti-metastatic effect of LPC relied on its regulation of gut microbiota such as increase of Lachnospiraceae UCG-006, Ruminococcus, and their metabolites such as acetic acid, propionic acid and butyric acid. In addition, LPC significantly inhibited CT26 colon cancer cells metastasis through increasing CD8+ cytotoxic T lymphocytes infiltration and decreasing the number of macrophages. Antibiotics treatment demonstrated that the therapeutic effect of LPC depended on the gut microbiota, which regulated T cells immune response. Taken together, LPC had strong inhibitory effects on colon cancer pulmonary metastasis by triggering gut-lung axis to influence the T cells immune response. Our research provides a novel finding for the utilization of procyanidins in the future, that is, supplementing more fruits and vegetables rich in procyanidins is beneficial to the treatment of colon cancer, or it can be used as an adjuvant drug in clinical anti-tumor immunotherapy.

A simple nanocomposite photocatalyst HT-rGO/TiO2 for deoxynivalenol degradation in liquid food.

A simple nanocomposite photocatalyst HT-rGO/TiO2 for deoxynivalenol (DON) degradation was synthesized by hydrothermal method to maintain the quality of cereal grains and byproducts. The characterization of HT-rGO/TiO2 was analyzed by XRD, FTIR, Raman spectroscopy, and XPS. Moreover, according to UV-vis DRS analysis, HT-rGO/TiO2 had a smaller band gap, indicating a wider response range to light and a higher utilized rate of quantum photons. Additionally, the results of LC-MS showed that the hydroxyl group at the C3 position, and the unsaturated bond between C9 and C10, and the epoxy group at C12 and C13 positions of DON molecule were destroyed step by step by photocatalytic degradation. These groups have active effects on the DON toxicity, which means it is successful to degrade DON in liquid-food by HT-rGO/TiO2 photocatalyst.

CRISPR-Cas-based detection for food safety problems: Current status, challenges, and opportunities.

Food safety is one of the biggest public issues occurring around the world. Microbiological, chemical, and physical hazards can lead to food safety issues, which may occur at all stages of the supply chain. In order to tackle food safety issues and safeguard consumer health, rapid, accurate, specific, and field-deployable detection methods meeting diverse requirements are one of the imperative measures for food safety assurance. CRISPR-Cas system, a newly emerging technology, has been successfully repurposed in biosensing and has demonstrated huge potential to establish conceptually novel detection methods with high sensitivity and specificity. This review focuses on CRISPR-Cas-based detection and its current status and huge potential specifically for food safety inspection. We firstly illustrate the pending problems in food safety and summarize the popular detection methods. We then describe the potential applications of CRISPR-Cas-based detection in food safety inspection. Finally, the challenges and futuristic opportunities are proposed and discussed. Generally speaking, the current food safety detection methods are still unsatisfactory in some ways such as being time-consuming, displaying unmet sensitivity and specificity standards, and there is a comparative paucity of multiplexed testing and POCT. Recent studies have shown that CRISPR-Cas-based biosensing is an innovative and fast-expanding technology, which could make up for the shortcomings of the existing methods or even replace them. To sum up, the implementation of CRISPR-Cas and the integration of CRISPR-Cas with other techniques is promising and desirable, which is expected to provide "customized" and "smart" detection methods for food safety inspection in the coming future.

Ghrelin ameliorates transformation of hepatic ischemia-reperfusion injury to liver fibrosis by blocking Smad and ERK signalling pathways, and promoting anti-inflammation and anti-oxidation effects.

BACKGROUND & AIMS: Ghrelin, a gut hormone with pleiotropic effects, may act as a protective signal in parenchymal cells. Hepatic ischemia-reperfusion injury (HIRI) causes acute-on-chronic liver failure and induces transformation of acute to chronic injury. HIRI model of mice was established by a semi-hepatic blocking method and treated with Ghrelin. This process is involved in inflammation, oxidative stress damage and apoptosis, and is associated with the expansion and activation of fibrotic haematopoietic stem cells (HSCs) which express and secrete high levels of collagen that induces liver fibrosis. Therefore, we investigated the effects of Ghrelin during transformation of HIRI to liver fibrosis, and explored the molecular mechanism of Ghrelin's action based on Smad and ERK pathways. METHODS: Hepatic injury was detected by plasma ALT levels. The hepatic histology and collagen were elucidated by HE staining and Masson staining, respectively. Liver inflammation levels and inflammatory cell counts were assessed by MPO and HE staining, respectively. The antioxidant capacity of plasma was evaluated based on the levels of SOD, MDA, and XOD. The mRNA or protein expression levels of genes related to apoptosis, fibrosis, Smad, and ERK pathways were assessed by real-time quantitative PCR (RT-qPCR), ELISA, or western blotting. RESULTS: The HIRI model was established to investigate the effects of the liver injury transformed to liver fibrosis. Ghrelin exhibited good hepatic protection by ameliorating liver histological changes and decreasing plasma ALT levels. Ghrelin significantly decreased the expression of MPO than that in model group, suggesting that Ghrelin blocked the inflammatory response in the HIRI liver tissue; this supports the anti-inflammatory effects of Ghrelin. Ghrelin significantly decreased apoptosis (enhanced Bcl-2 expression, and down-regulated Bax and Caspase 3). Ghrelin exhibited anti-oxidative effects as it inhibited plasma MDA levels, and promoted plasma SOD and XOD levels. Moreover, Ghrelin inhibited activation of hepatic stellate cells, blocked traditional fibrotic Smad and ERK signalling pathways, and reduced hepatic fibrosis by stimulating degradation of extracellular matrices (ECMs; such as collagen I, collagen III, HA, and LN). CONCLUSIONS: This study demonstrates that Ghrelin delays the transformation of HIRI to liver fibrosis process which is correlated to its anti-apoptotic, anti-inflammatory, and anti-oxidative effects. Moreover, Ghrelin alleviates HIRI-mediated liver fibrosis, inhibits activation of HSCs, and reduces accumulation of ECM via inhibition of Smad and ERK signalling pathways.

A smartphone-based visual biosensor for CRISPR-Cas powered SARS-CoV-2 diagnostics.

The pandemic of coronavirus disease 2019 (COVID-19) resulted from novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become a worldwide concern. It is imperative to develop rapid, sensitive, and specific biosensing methods. Herein, we developed a CRISPR-Cas12a powered visual biosensor with a smartphone readout for ultrasensitive and selective detection of SARS-CoV-2. Simply, the SARS-CoV-2 derived nucleic acids triggered CRISPR-Cas12a based indiscriminate degradation of a single-stranded DNA that was supposed to link two gold nanoparticles, inducing the dis-aggregation of gold nanoparticles and thus generating observable color changes. This change can be readily distinguished by naked eyes as well as a smartphone with a Color Picker App. The proposed biosensor was successfully applied to detect SARS-CoV-2 gene in synthetic vectors, transcribed RNA and SARS-CoV-2 pseudoviruses. It rendered "single copy resolution" as evidenced by the 1 copy/µL limit of detection of pseudoviruses with no cross-reactivity. When the developed biosensor was challenged with SARS-CoV-2 clinical bio-samples, it provided 100% agreement (both positive and negative) with qPCR results. The sample-to-result time was roughly 90 min. Our work provides a novel and robust technology for ultrasensitive detection of SARS-CoV-2 that could be used clinically.

A novel Ag2O-TiO2-Bi2WO6/polyvinyl alcohol composite film with ethylene photocatalytic degradation performance towards banana preservation.

In this research, the Ag2O-TiO2-Bi2WO6(ATB) ternary heterojunction photocatalyst was synthesized by hydrothermal and surface deposition method, and the ATB/PVA composite film with ethylene photocatalytic degradation performance was constructed by the casting method. The structure and properties of ATB and ATB/PVA films were characterized and applied to banana preservation. The results showed that the addition of ATB could improve the mechanical properties, thermal stability, oxygen and moisture resistance, and reduce the crystallinity and light transmittance of PVA films. Compared with TiO2, Bi2WO6 and TB photocatalysts, ATB had the best photocatalytic degradation effect of ethylene under LED light. Compared with blank group, the ethylene concentration decreased by 17.17%. This was mainly attributed to the formation of heterostructure among Ag2O, TiO2 and Bi2WO6, which promoted the separation and transfer of photogenerated carriers. The ATB/PVA composite coating could effectively prevent the respiration of the bananas by inhibiting gas exchange and degrading ethylene, which reduced the weight loss, inhibited glycogen decomposition, improved the pulp hardness, increased titratable acid content, reduced the PPO activity, hindered the phenol oxidation and keep better apparent color of bananas. The safety study suggested that the ATB/PVA film is safe for bananas packaging application.

CRISPR-Cas based virus detection: Recent advances and perspectives.

Viral infections are one of the most intimidating threats to human beings. One convincing example is the coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2. Rapid, sensitive, specific and field-deployable identification of causal viruses is critical for disease surveillance, control and treatment. The shortcomings of current methods create an impending need for developing novel biosensing platforms. CRISPR-Cas systems, especially CRISPR-Cas12a and CRISPR-Cas13a, characterized by their sensitivity, specificity, high base resolution and programmability upon nucleic acid recognition, have been repurposed for molecular diagnostics, surging a new path forward in biosensing. They, as the core of some robust diagnostic tools, are revolutionizing the way that virus can be detected. This review focuses on recent advances in virus detection with CRISPR-Cas systems especially CRISPR-Cas12a/Cas13a. We started with a short introduction to CRISPR-Cas systems and the properties of Cas12a and Cas13a effectors, and continued with reviewing the current advances of virus detection utilizing CRISPR-Cas systems. The significance and advantages of such methods were then discussed. Finally, the challenges and perspectives were proposed. We tried to provide readers with a concise profile of emerging and fast-expanding CRISPR-Cas based biosensing technology, and highlighted its potential applications in a range of scenarios with regard to virus detection.

Polyphenol-Rich Extract from Litchi chinensis Seeds Alleviates Hypertension-Induced Renal Damage in Rats.

Litchi chinensis seed is a valuable byproduct of the subtropical fruit litchi (L. chinensis Sonn.), whose extract (LSE) has been confirmed to ameliorate dyslipidemia, hyperglycemia, and oxidative stress caused by type 2 diabetes. However, if LSE exerts an effect on anti-hypertension and hypertensive renal damage remains unknown. In this study, 13 polyphenols and one fatty acid were identified by UPLC-Q/TOF-MS. Network pharmacological analysis revealed that the therapeutic effects of LSE may be involved in multitargets and multipathways, such as the TNF signaling pathway, interleukin (IL)-6-mediated signaling pathway, NF-kappa B signaling pathway, removal of superoxide radicals, negative regulation of blood pressure, and so forth. Moreover, spontaneously hypertensive rats (SHRs) were daily gavaged with LSE (60 mg/kg) for 10 weeks. LSE remarkably reduced systolic blood pressure (SBP). The hypertension-induced renal damage was improved by suppressing inflammation and oxidative stress, which was consistent with the prediction of network pharmacology. In addition, LSE treatment remarkably increased the relative abundances of Lactobacillus and the production of short-chain fatty acids in the intestine. Our study indicated that a byproduct of litchi, namely, litchi seed, may be effective in reducing SBP and alleviating hypertensive renal damage.

Novel visible light-responsive graphene oxide/Bi2WO6/starch composite membrane for efficient degradation of ethylene.

Graphene oxide/Bi2WO6 (GBW) photocatalyst was synthesized using a hydrothermal and surface deposition method. GBW/starch composite films with different graphene oxide (GO) additions (0, 0.25, 0.5, 0.75, 1 %) were prepared using a casting method. The GBW photocatalyst and composite starch film were characterized using X-ray diffractometry, X-ray photoelectron spectroscopy, Ultraviolet-visible diffuse reflectance spectroscopy, scanning electron microscopy, Fourier transform infrared spectroscopy, synchronous thermal analyzer, and the capacity of photocatalytic degradation of ethylene under visible light was evaluated. The results showed that GBW strengthens the mechanical properties, water vapor resistance and thermal stability of the composite film. Proper introduction of GO can refine lattice size, reduce bandgap and enhance visible light absorption. When the addition of GO was 0.5 %, GBW/starch composite film showed the strongest visible light degradation activity for ethylene, and the rate constant K' was 9.91 × 10-4 min-1, 4.4 times that of pure Bi2WO6. The composite film also had good recycling performance.

Construction of nanoscale liposomes loaded with melatonin via supercritical fluid technology.

Melatonin-loaded liposomes (MLL) were successfully prepared using rapid expansion of supercritical solution technology. The effects of supercritical pressure on encapsulation efficiency (EE) and average particle size were then analysed. Meanwhile, temperature, formation time and ethanol concentration in the products were studied and optimised based on the response surface methodology (RSM). An in vitro simulated digestion model was also established to evaluate the release performance of MLL. The results showed that 140 bar was the best pressure for maximising the EE value using RSM optimisation, reaching up to 82.2%. MLL characterisations were performed using analytic techniques including infrared spectroscopy, transmission electron microscopy, a laser scattering particle size analyser and gas chromatograph-mass spectrometer. The size distribution was uniform, with an average diameter of 66 nm. Stability tests proved that MLL maintained good preservation duration, and residual solvent experiments indicated that only 1.03% (mass ratio) of ethanol remained in the products. Simulated release experiments indicated the slow release feature in early digestive stages and more thorough characteristics in later stages of simulated digestion.

Enhanced photocatalytic disinfection of P. expansum in cold storage using a TiO2/ACF film.

Spores of Penicilliumexpansum are the most important airborne component of fungal contamination and are commonly present in the moist air in cold storage rooms where fruits and vegetables are stored. To improve the ability of activated carbon felt-supported titanium dioxide (TiO2/ACF) to remove spores of P. expansum from the atmosphere, measurements for the removal efficiency of the spores at a temperature of 3 degrees C+/-1 degrees C and relative humidity of 90%+/-3% had been made on photocatalytically-activated (PC) silver-doped TiO2/ACF prepared by ion sputtering in two different modes and photoelectrocatalytically-activated (PEC) TiO2/ACF films. The Weibull distribution model was used to define the degree of microbial inactivation attributable to the PC or PEC films. The key parameters of the PC or PEC affecting the disinfection efficiency in terms of the reliable life (t(R) value) of this model were studied. Both silver-doped TiO2/ACF and PEC on TiO2/ACF improved destruction of this airborne microbe. Silver deposited on the surface of the prepared ACF-supported TiO2 films (Ag/Ti=0.012, atomic ratios) dramatically reduced t(R) value. With respect to the PEC, a model was established using response surface methodology to describe the relationship between the t(R) value and the key affecting parameters, including light intensity and bias voltage. The optimized parameters were found to be a light intensity of 2.3 mW x cm(-2) with a bias voltage of 66.7 V.

Hydroxysafflor yellow A protects neuron against hypoxia injury and suppresses inflammatory responses following focal ischemia reperfusion in rats.

Previous data demonstrated that hydroxysafflor yellow A (HSYA), a yellow color pigments extracted from the safflower, was an effective agent against focal cerebral ischemia. In the present study we demonstrated that HSYA prevented the injury in cultured cerebral cortical neurons induced by oxygen-glucose deprivation and increased the cell viability, as shown by the inhibition of both LDH and NO efflux. Further, HSYA administered orally 3 d before middle cerebral artery occlusion has the capacity to reduce cerebral infarct size and edema after 2 h cerebral ischemia followed by 24 h reperfusion in rats, and to significantly improve neurological behavior scores. Mean while, treatment with HSYA significantly decreased both mRNA and protein levels of IL-1beta, TNF-alpha in ischemic brain tissue. These results suggested that the protection of HSYA results from, at least in part, suppression of inflammatory responses following focal ischemia reperfusion.