Drip-Dry Strategy Assisted Blu-Ray Disc Biosensor for Fast Point of Care Testing.

Discs and numerous other consumer products have been developed for point of care testing (POCT) to replace traditional large and expensive biochemical devices in certain scenarios. Herein, we propose a drip-dry strategy (2D strategy) assisted Blu-ray disc (BD) biosensor, termed BDB, for rapid and portable POCT within 30 min with the cost of a single test < $1. The platform utilizes the covered area formed by the deposition of the substance to be measured on the activated BD surface after the evaporation of water and realizes the quantitative detection of the target through the error readout of free disc quality diagnosis software. As a proof of concept, we first demonstrated the feasibility of direct quantitative detection of substances in solution in a single system through the detection of pure proteins avoiding colorimetric reagent used in traditional optical detection. For the complex mixed systems, we then innovatively utilize the principle that soluble targets promote/inhibit the dissolution of insoluble precipitates to achieve specific detection of targets and successfully apply BDB to the indirect quantitative detection of glutathione (GSH) with LOD of 0.447 mM in the range of 2-16 mM and organophosphorus pesticides (OPs) with LOD of 2.122 × 10-7 M in the range of 1.289 × 10-7-1.289 × 10-4 M. The BDB is widely applicable, easy to operate, and less time-consuming, which is anticipated to provide an alternative method for early, on-site detection or screening.

Advanced Palladium Nanosheet-Enhanced Phototherapy for Treating Wound Infection Caused by Multidrug-Resistant Bacteria.

With the increasing spread of multidrug-resistant (MDR) bacteria worldwide, it is needed to develop antibiotics-alternative strategies for the treatment of bacterial infections. This work develops a multifunctional single-component palladium nanosheet (PdNS) with broad-spectrum and highly effective bactericidal activity against MDR bacteria. PdNS exerts its endogenous nanoknife (mechanical cutting) effect and peroxidase-like activity independent of light. Under near-infrared region (NIR) light irradiation, PdNS exhibits photothermal effect to produce local heat and meanwhile possesses photodynamic effect to generate 1O2; notably, PdNS has catalase-like activity-dependent extra photodynamic effect upon H2O2 addition. PdNS+H2O2+NIR employs a collectively synergistic mechanism of nanoknife effect, peroxidase/catalase-like catalytic activity, photothermal effect, and photodynamic effect for bacterial killing. PdNS+H2O2+NIR causes compensatory elevated phospholipid biosynthesis, disordered energy metabolism, increased cellular ROS levels and excessive oxidative stress, and inhibited nucleic acid synthesis in bacteria. In mice, PdNS+H2O2+NIR gives >92.7% bactericidal rates at infected wounds and almost the full recovery of infected wounds, and it leads to extensive down-regulation of proinflammatory pathways and comprehensive up-regulation of wound healing pathways, conferring elevated inflammation resolution and meanwhile accelerated wound repair. PdNS+H2O2+NIR represents a highly efficient nanoplatform for photoenhanced treatment of superficial infections.

Fully Integrated and High-Throughput Microfluidic System for Multiplexed Point-Of-Care Testing.

For every epidemic outbreak, the prevention and treatments in resource-limited areas are always out of reach. Critical to this is that high accuracy, stability, and more comprehensive analytical techniques always rely on expensive and bulky instruments and large laboratories. Here, a fully integrated and high-throughput microfluidic system is proposed for ultra-multiple point-of-care immunoassay, termed Dac system. Specifically, the Dac system only requires a handheld portable device to automatically recycle repetitive multi-step reactions including on-demand liquid releasing, dispensing, metering, collecting, oscillatory mixing, and discharging. The Dac system performs high-precision enzyme-linked immunosorbent assays for up to 17 samples or targets simultaneously on a single chip. Furthermore, reagent consumption is only 2% compared to conventional ELISA, and microbubble-accelerated reactions shorten the assay time by more than half. As a proof of concept, the multiplexed detections are achieved by detecting at least four infection targets for two samples simultaneously on a singular chip. Furthermore, the barcode-based multi-target results can rapidly distinguish between five similar cases, allowing for accurate therapeutic interventions. Compared to bulky clinical instruments, the accuracy of clinical inflammation classification is 92.38% (n = 105), with a quantitative correlation coefficient of R2 = 0.9838, while the clinical specificity is 100% and the sensitivity is 98.93%.

Comparative pathogenicity of influenza virus-induced pneumonia mouse model following intranasal and aerosolized intratracheal inoculation.

BACKGROUND: Infection of mice with mouse-adapted strains of influenza virus has been widely used to establish mouse pneumonia models. Intranasal inoculation is the traditional route for constructing an influenza virus-induced pneumonia mouse model, while intratracheal inoculation has been gradually applied in recent years. In this article, the pathogenicity of influenza virus-induced pneumonia mouse models following intranasal and aerosolized intratracheal inoculation were compared. METHODS: By comparing the two ways of influenza inoculation, intranasal and intratracheal, a variety of indices such as survival rate, body weight change, viral titer and load, pathological change, lung wet/dry ratio, and inflammatory factors were investigated. Meanwhile, the transcriptome was applied for the initial exploration of the mechanism underlying the variations in the results between the two inoculation methods. RESULTS: The findings suggest that aerosolized intratracheal infection leads to more severe lung injury and higher viral loads in the lungs compared to intranasal infection, which may be influenced by the initial site of infection, sialic acid receptor distribution, and host innate immunity. CONCLUSION: Intratracheal inoculation is a better method for modelling severe pneumonia in mice than intranasal infection.

A new method for the joint estimation of instantaneous reproductive number and serial interval during epidemics.

Although some methods for estimating the instantaneous reproductive number during epidemics have been developed, the existing frameworks usually require information on the distribution of the serial interval and/or additional contact tracing data. However, in the case of outbreaks of emerging infectious diseases with an unknown natural history or undetermined characteristics, the serial interval and/or contact tracing data are often not available, resulting in inaccurate estimates for this quantity. In the present study, a new framework was specifically designed for joint estimates of the instantaneous reproductive number and serial interval. Concretely, a likelihood function for the two quantities was first introduced. Then, the instantaneous reproductive number and the serial interval were modeled parametrically as a function of time using the interpolation method and a known traditional distribution, respectively. Using the Bayesian information criterion and the Markov Chain Monte Carlo method, we ultimately obtained their estimates and distribution. The simulation study revealed that our estimates of the two quantities were consistent with the ground truth. Seven data sets of historical epidemics were considered and further verified the robust performance of our method. Therefore, to some extent, even if we know only the daily incidence, our method can accurately estimate the instantaneous reproductive number and serial interval to provide crucial information for policymakers to design appropriate prevention and control interventions during epidemics.

Combining early post-resuscitation EEG and HRV features improves the prognostic performance in cardiac arrest model of rats.

OBJECTIVE: Early and reliable prediction of neurological outcome remains a challenge for comatose survivors of cardiac arrest (CA). The purpose of this study was to evaluate the predictive ability of EEG, heart rate variability (HRV) features and the combination of them for outcome prognostication in CA model of rats. METHODS: Forty-eight male Sprague-Dawley rats were randomized into 6 groups (n=8 each) with different cause and duration of untreated arrest. Cardiopulmonary resuscitation was initiated after 5, 6 and 7min of ventricular fibrillation or 4, 6 and 8min of asphyxia. EEG and ECG were continuously recorded for 4h under normothermia after resuscitation. The relationships between features of early post-resuscitation EEG, HRV and 96-hour outcome were investigated. Prognostic performances were evaluated using the area under receiver operating characteristic curve (AUC). RESULTS: All of the animals were successfully resuscitated and 27 of them survived to 96h. Weighted-permutation entropy (WPE) and normalized high frequency (nHF) outperformed other EEG and HRV features for the prediction of survival. The AUC of WPE was markedly higher than that of nHF (0.892 vs. 0.759, p<0.001). The AUC was 0.954 when WPE and nHF were combined using a logistic regression model, which was significantly higher than the individual EEG (p=0.018) and HRV (p<0.001) features. CONCLUSIONS: Earlier post-resuscitation HRV provided prognostic information complementary to quantitative EEG in the CA model of rats. The combination of EEG and HRV features leads to improving performance of outcome prognostication compared to either EEG or HRV based features alone.

Comparison of Quantitative Characteristics of Early Post-resuscitation EEG Between Asphyxial and Ventricular Fibrillation Cardiac Arrest in Rats.

BACKGROUND: Quantitative electroencephalogram (EEG) analysis has shown promising results in studying brain injury and functional recovery after cardiac arrest (CA). However, whether the quantitative characteristics of EEG, as potential indicators of neurological prognosis, are influenced by CA causes is unknown. The purpose of this study was designed to compare the quantitative characteristics of early post-resuscitation EEG between asphyxial CA (ACA) and ventricular fibrillation CA (VFCA) in rats. METHODS: Thirty-two Sprague-Dawley rats of both sexes were randomized into either ACA or VFCA group. Cardiopulmonary resuscitation was initiated after 5-min untreated CA. Characteristics of early post-resuscitation EEG were compared, and the relationships between quantitative EEG features and neurological outcomes were investigated. RESULTS: Compared with VFCA, serum level of S100B, neurological deficit score and brain histopathologic damage score were dramatically higher in the ACA group. Quantitative measures of EEG, including onset time of EEG burst, time to normal trace, burst suppression ratio, and information quantity, were significantly lower for CA caused by asphyxia and correlated with the 96-h neurological outcome and survival. CONCLUSIONS: Characteristics of earlier post-resuscitation EEG differed between cardiac and respiratory causes. Quantitative measures of EEG not only predicted neurological outcome and survival, but also have the potential to stratify CA with different causes.

Simulating the potential role of media coverage and infected bats in the 2014 Ebola outbreak.

Multiple epidemiological models have been developed to model the transmission dynamics of Ebola virus (EBOV) disease in West Africa in 2014 because the severity of the epidemic is commonly overestimated. A compartmental model that incorporates the media impact and the effect of infected bats was constructed and calibrated using data reported until the end of 2014. The final cumulative number of deaths and confirmed cases were estimated to be 1.0921×104 (95% CI 9.7706×103-1.2072×104) and 1.5193×104 (95% CI 1.3593×104-1.6795×104), respectively. The epidemic was estimated to end on June 2015, which was similar to the data reported by the World Health Organization. A sensitivity analysis indicated that an increase of either the media impact or the number of infectious bats that are captured daily can increase the cumulative number of confirmed cases/deaths. Of the considered epidemiological parameters, only the media coverage can significantly reduce both the peak time and the value of the cumulative confirmed cases/deaths. Thus, we propose 'the cumulative confirmed cases and deaths' as another media mechanism. In conclusion, the media impact contributed to the control of the 2014 Ebola outbreak, and infectious bats may be a potential source of the epidemic.

An Effective Terrain Aided Navigation for Low-Cost Autonomous Underwater Vehicles.

Terrain-aided navigation is a potentially powerful solution for obtaining submerged position fixes for autonomous underwater vehicles. The application of terrain-aided navigation with high-accuracy inertial navigation systems has demonstrated meter-level navigation accuracy in sea trials. However, available sensors may be limited depending on the type of the mission. Such limitations, especially for low-grade navigation sensors, not only degrade the accuracy of traditional navigation systems, but further impact the ability to successfully employ terrain-aided navigation. To address this problem, a tightly-coupled navigation is presented to successfully estimate the critical sensor errors by incorporating raw sensor data directly into an augmented navigation system. Furthermore, three-dimensional distance errors are calculated, providing measurement updates through the particle filter for absolute and bounded position error. The development of the terrain aided navigation system is elaborated for a vehicle equipped with a non-inertial-grade strapdown inertial navigation system, a 4-beam Doppler Velocity Log range sensor and a sonar altimeter. Using experimental data for navigation performance evaluation in areas with different terrain characteristics, the experiment results further show that the proposed method can be successfully applied to the low-cost AUVs and significantly improves navigation performance.

Unraveling host-pathogen dynamics in a murine Model of septic peritonitis induced by vancomycin-resistant Enterococcus faecium.

Vancomycin-resistant Enterococcus faecium (E. faecium) infection is associated with higher mortality rates. Previous studies have emphasized the importance of innate immune cells and signalling pathways in clearing E. faecium, but a comprehensive analysis of host-pathogen interactions is lacking. Here, we investigated the interplay of host and E. faecium in a murine model of septic peritonitis. Following injection with a sublethal dose, we observed significantly increased murine sepsis score and histological score, decreased weight and bacterial burden, neutrophils and macrophages infiltration, and comprehensive activation of cytokine-mediated signalling pathway. In mice receiving a lethal dose, hypothermia significantly improved survival, reduced bacterial burden, cytokines, and CD86 expression of MHC-II+ recruited macrophages compared to the normothermia group. A mathematical model constructed by observational data from 80 animals, recapitulated the host-pathogen interplay, and further verified the benefits of hypothermia. These findings indicate that E. faecium triggers a severe activation of cytokine-mediated signalling pathway, and hypothermia can improve outcomes by reducing bacterial burden and inflammation.

Fully integrated and automated centrifugal microfluidic chip for point-of-care multiplexed molecular diagnostics.

Public health events caused by pathogens have imposed significant economic and societal burdens. However, conventional methods still face challenges including complex operations, the need for trained operators, and sophisticated instruments. Here, we proposed a fully integrated and automated centrifugal microfluidic chip, also termed IACMC, for point-of-care multiplexed molecular diagnostics by harnessing the advantages of active and passive valves. The IACMC incorporates multiple essential components including a pneumatic balance module for sequential release of multiple reagents, a pneumatic centrifugation-assisted module for on-demand solution release, an on-chip silicon membrane module for nucleic acid extraction, a Coriolis force-mediated fluid switching module, and an amplification module. Numerical simulation and visual validation were employed to iterate and optimize the chip's structure. Upon sample loading, the chip automatically executes the entire process of bacterial sample lysis, nucleic acid capture, elution quantification, and isothermal LAMP amplification. By optimizing crucial parameters including centrifugation speed, direction of rotation, and silicone membrane thickness, the chip achieves exceptional sensitivity (twenty-five Salmonella or forty Escherichia coli) and specificity in detecting Escherichia coli and Salmonella within 40 min. The development of IACMC will drive advancements in centrifugal microfluidics for point-of-care testing and holds potential for broader applications in precision medicine including high-throughput biochemical analysis immune diagnostics, and drug susceptibility testing.

Real-time amplitude spectrum area estimation during chest compression from the ECG waveform using a 1D convolutional neural network.

Introduction: Amplitude spectrum area (AMSA) is a well-established measure than can predict defibrillation outcome and guiding individualized resuscitation of ventricular fibrillation (VF) patients. However, accurate AMSA can only be calculated during cardiopulmonary resuscitation (CPR) pause due to artifacts produced by chest compression (CC). In this study, we developed a real-time AMSA estimation algorithm using a convolutional neural network (CNN). Methods: Data were collected from 698 patients, and the AMSA calculated from the uncorrupted signals served as the true value for both uncorrupted and the adjacent corrupted signals. An architecture consisting of a 6-layer 1D CNN and 3 fully connected layers was developed for AMSA estimation. A 5-fold cross-validation procedure was used to train, validate and optimize the algorithm. An independent testing set comprised of simulated data, real-life CC corrupted data, and preshock data was used to evaluate the performance. Results: The mean absolute error, root mean square error, percentage root mean square difference and correlation coefficient were 2.182/1.951 mVHz, 2.957/2.574 mVHz, 22.887/28.649% and 0.804/0.888 for simulated and real-life testing data, respectively. The area under the receiver operating characteristic curve regarding predicting defibrillation success was 0.835, which was comparable to that of 0.849 using the true value of the AMSA. Conclusions: AMSA can be accurately estimated during uninterrupted CPR using the proposed method.

Manganese-Based Nanoparticle Vaccine for Combating Fatal Bacterial Pneumonia.

Bacterial pneumonia is the leading cause of death worldwide among all infectious diseases. However, currently available vaccines against fatal bacterial lung infections, e.g., pneumonic plague, are accompanied by limitations, including insufficient antigen-adjuvant co-delivery and inadequate immune stimulation. Therefore, there is an urgent requirement to develop next-generation vaccines to improve the interaction between antigen and adjuvant, as well as enhance the effects of immune stimulation. This study develops a novel amino-decorated mesoporous manganese silicate nanoparticle (AMMSN) loaded with rF1-V10 (rF1-V10@AMMSN) to prevent pneumonic plague. These results suggest that subcutaneous immunization with rF1-V10@AMMSN in a prime-boost strategy induces robust production of rF1-V10-specific IgG antibodies with a geometric mean titer of 315,844 at day 42 post-primary immunization, which confers complete protection to mice against 50 × LD50 of Yersinia pestis (Y. pestis) challenge via the aerosolized intratracheal route. Mechanistically, rF1-V10@AMMSN can be taken up by dendritic cells (DCs) and promote DCs maturation through activation of the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway and production of type I interferon. This process results in enhanced antigen presentation and promotes rF1-V10-mediated protection against Y. pestis infection. This manganese-based nanoparticle vaccine represents a valuable strategy for combating fatal bacterial pneumonia.

Adaptive Weighted Neighbors Method for Sensitivity Analysis.

Identifying key factors from observational data is important for understanding complex phenomena in many disciplines, including biomedical sciences and biology. However, there are still some limitations in practical applications, such as severely nonlinear input-output relationships and highly skewed output distributions. To acquire more reliable sensitivity analysis (SA) results in these extreme cases, inspired by the weighted k-nearest neighbors algorithm, we propose a new method called adaptive weighted neighbors (AWN). AWN makes full use of the information contained in all training samples instead of limited samples and automatically gives more weight to nearby samples. Then, the bootstrap technique and Jansen's method are used to obtain reliable SA results based on AWN. We demonstrate the performance and accuracy of AWN by analyzing various biological and biomedical data sets, three simulated examples and two case studies, showing that it can effectively overcome the above limitations. We therefore expect it to be a complementary approach for SA.

Limonin relieves TGF-ß-induced hepatocyte EMT and hepatic stellate cell activation in vitro and CCl4-induced liver fibrosis in mice via upregulating Smad7 and subsequent suppression of TGF-ß/Smad cascade.

Liver fibrosis is a pathological process as a result of intrahepatic deposition of excessive ECM. EMT of hepatocytes and activation of HSCs both play important roles in the etiology of liver fibrosis. Here, we found that limonin repressed TGF-ß-induced EMT in AML-12 hepatocytes and activation of LX-2 HSCs. Limonin suppressed TGF-ß-provoked Smad2/3 C-terminal phosphorylation and subsequent nuclear translocation. However, limonin exerted few effects on Smad2/3 phosphorylation atlinker region. Mechanistically, limonin increased Smad7 in both AML-12 and LX-2 cells. Knockdown of Smad7 abrogated inhibitory effects of limonin on TGF-ß-induced changes in both two cells. Further studies revealed that limonin upregulated Smad7 and declined C-terminal phosphorylation and nuclear translocation of Smad2/3 to alleviate mouse CCl4-induced liver fibrosis. Our findings indicated that limonin inhibits TGF-ß-induced EMT of hepatocytes and activation of HSCs in vitro and CCl4-induced liver fibrosis in mice. Upregulated Smad7 which suppresses Smad2/3-dependent gene transcription is implicated in the hepatoprotective activity of limonin.

A switching dynamic model based on phased COVID-19 data in Chongqing and its evaluation.

OBJECTIVES: Although COVID-19 has been controlled in China, the risk of invasion of imported cases remains. We aimed to characterize the impact of the number of imported cases and the implementation of first-level emergency response (FLER) policy. METHODS: A SCQIHR switching model was constructed and verified by the complete phased data of COVID-19 in Chongqing in 2020. Then it was used to investigate the impact of the number of imported cases and the timing of FLER. Lastly, it was evaluated by three actual scenarios in Chongqing in 2021. RESULTS: The proposed model can fit the multidimensional time series well. After the implementation of FLER, the mean effective reproduction number, contact rate and misdetection rate were decreased significantly, but the quarantine rate for close contacts and isolation rate for non-hospitalized infectious cases were increased significantly. The peaks of quarantined close contacts and hospitalized infectious cases increased linearly with the increase of the number of imported cases and the lag of FLER time, which was verified by three actual scenarios in Chongqing in 2021. CONCLUSIONS: These findings can provide guidance for local public health policy-making and allocation of medical resources, reduce the impact of COVID-19 on the local population.

Trimetazidine Alleviates Postresuscitation Myocardial Dysfunction and Improves 96-Hour Survival in a Ventricular Fibrillation Rat Model.

Background Myocardial dysfunction is a critical cause of post-cardiac arrest hemodynamic instability and circulatory failure that may lead to early mortality after resuscitation. Trimetazidine is a metabolic agent that has been demonstrated to provide protective effects in myocardial ischemia. However, whether trimetazidine protects against postresuscitation myocardial dysfunction is unknown. Methods and Results Cardiopulmonary resuscitation was initiated after 8 minutes of untreated ventricular fibrillation in Sprague-Dawley rats. Animals were randomized to 4 groups immediately after resuscitation (n=15/group): (1) normothermia control (NTC); (2) targeted temperature management; (3) trimetazidine-normothermia; (4) trimetazidine-targeted temperature management. TMZ was administered at a single dose of 10 mg/kg in rats with trimetazidine. The body temperature was maintained at 34.0°C for 2 hours and then rewarmed to 37.5°C in rats with targeted temperature management. Postresuscitation hemodynamics, 96-hours survival, and pathological analysis were assessed. Heart tissues and blood samples of additional rats (n=6/group) undergoing the same experimental procedure were collected to measure myocardial injury, inflammation and oxidative stress-related biomarkers with ELISA-based quantification assays. Compared with normothermia control, tumor necrosis factor-α, and cardiac troponin-I were significantly reduced, whereas the left ventricular ejection fraction and 96-hours survival rates were significantly improved in the 3 experimental groups. Furthermore, inflammation and oxidative stress-related biomarkers together with collagen volume fraction were significantly decreased in rats undergoing postresuscitation interventions. Conclusions Trimetazidine significantly alleviates postresuscitation myocardial dysfunction and improves survival by decreasing oxidative stress and inflammation in a ventricular fibrillation rat model. A single dose of trimetazidine administrated immediately after resuscitation can effectively improve cardiac function, whether used alone or combined with targeted temperature management.

A Dynamic Compartmental Model to Explore the Optimal Strategy of Varicella Vaccination: An Epidemiological Study in Jiangsu Province, China.

Varicella (chickenpox) is highly contagious among children and frequently breaks out in schools. In this study, we developed a dynamic compartment model to explore the optimal schedule for varicella vaccination in Jiangsu Province, China. A susceptible-infected-recovered (SIR) model was proposed to simulate the transmission of varicella in different age groups. The basic reproduction number was computed by the kinetic model, and the impact of three prevention factors was assessed through the global sensitivity analysis. Finally, the effect of various vaccination scenarios was qualitatively evaluated by numerical simulation. The estimated basic reproduction number was 1.831 ± 0.078, and the greatest contributor was the 5-10 year-old group (0.747 ± 0.042, 40.80%). Sensitivity analysis indicated that there was a strong negative correlation between the second dose vaccination coverage rate and basic reproduction number. In addition, we qualitatively found that the incidence would significantly decrease as the second dose vaccine coverage expands. The results suggest that two-dose varicella vaccination should be mandatory, and the optimal age of second dose vaccination is the 5-10 year-old group. Optimal vaccination time, wide vaccine coverage along with other measures, could enhance the effectiveness of prevention and control of varicella in China.

A characterized saponin extract of Panax japonicus suppresses hepatocyte EMT and HSC activation in vitro and CCl4-provoked liver fibrosis in mice: Roles of its modulatory effects on the Akt/GSK3ß/Nrf2 cascade.

BACKGROUND AND PURPOSE: Liver fibrosis constitutes a pathologic condition resulting in a series of advanced liver diseases. Oleanane-type saponins are distinctive active constituents in the medicinal plant Panax japonicus C. A. Mey (P. japonicus). Herein, we assessed protective effects of a characterized saponin extract of rhizomes of P. japonicus (SEPJ) on hepatocyte EMT and HSC activation in vitro and liver fibrosis in mice. We also investigated molecular mechanisms underlying the hepatoprotective activity of SEPJ. METHODS: EMT of AML-12 hepatocytes was evaluated by observing morphology of cells and quantifying EMT marker proteins. Activation of LX-2 HSCs was assessed via scratch assay, transwell assay, and EdU-incorporation assay, and by quantifying activation marker proteins. Liver fibrosis in mice was evaluated by HE, SR, and Masson staining, and by measuring related serum indicators. Immunoblotting and RT-PCR were performed to study mechanisms underlying the action of SEPJ. RESULTS: SEPJ inhibited TGF-ß-induced EMT in AML-12 hepatocytes and activation of LX-2 HSCs. SEPJ elevated Akt phosphorylation at Ser473 and GSK3ß phosphorylation at Ser9 in these cells, giving rise to a descent of the catalytic activity of GSK3ß. These events increased levels of both total and nuclear Nrf2 protein and upregulated expressions of Nrf2-responsive antioxidative genes. In addition, enhanced phosphorylation of Akt and GSK3ß acted upstream of SEPJ-mediated activation of Nrf2. Knockdown of Nrf2 or inhibition of Akt diminished the protective activity of SEPJ against TGF-ß in both AML-12 and LX-2 cells. Our further in vivo experiments revealed that SEPJ imposed a considerable alleviation on CCl4-provoked mouse liver fibrosis. Moreover, hepatic Akt/GSK3ß/Nrf2 cascade were potentiated by SEPJ. Taken together, our results unveiled that SEPJ exerted protective effects against fibrogenic cytokine TGF-ß in vitro and ameliorated liver fibrosis in mice. Mechanistically, SEPJ regulated the Akt/GSK3ß/Nrf2 signaling which subsequently enhanced intracellular antioxidative capacity. CONCLUSIONS: SEPJ inhibits hepatocyte EMT and HSC activation in vitro and alleviates liver fibrosis in mice. Modulation of the Akt/GSK3ß/Nrf2 cascade attributes to its hepatoprotective effects. Our findings support a possible application of SEPJ in the control of liver fibrosis.

Piperine inhibits AML-12 hepatocyte EMT and LX-2 HSC activation and alleviates mouse liver fibrosis provoked by CCl4: roles in the activation of the Nrf2 cascade and subsequent suppression of the TGF-ß1/Smad axis.

Piperine (PIP) is an alkaloid derived from peppercorns. Herein, we assessed its effects on hepatocyte EMT and HSC activation in vitro and CCl4-elicited liver fibrosis in mice. Further experiments were performed to unveil the molecular mechanisms underlying the hepatoprotective activity of PIP. We found that PIP inhibited TGF-ß1-provoked AML-12 hepatocyte EMT and LX-2 HSC activation. Mechanistically, in AML-12 and LX-2 cells, PIP evoked Nrf2 nuclear translocation and increased transcriptions of Nrf2-responsive antioxidative genes. These events decreased TGF-ß1-induced production of ROS. Moreover, PIP increased the expression of Smad7, suppressed phosphorylation and nuclear translocation of Smad2/3, and decreased the transcriptions of Smad2/3-downstream genes. Knockdown of Nrf2 abrogated the protective activity of PIP against TGF-ß1. Modulatory effects of PIP on the TGF-ß1/Smad cascade were also crippled, which suggested that activation of Nrf2 played critical roles in the regulatory effects of PIP on TGF-ß1/Smad signaling. Experiments in vivo unveiled that PIP ameliorated mouse liver fibrosis provoked by CCl4. PIP modulated the intrahepatic contents of the markers of EMT and HSC activation. In mouse livers, PIP activated Nrf2 signaling and reduced Smad2/3-dependent gene transcriptions. Our findings collectively suggested PIP as a new chemical entity with the capacity of alleviating liver fibrosis. The activation of the Nrf2 cascade and subsequent suppression of the TGF-ß1/Smad axis are implicated in the hepatoprotective activity of PIP.