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The abundance of molecules on early Earth likely enabled a wide range of prebiotic chemistry, with peptides playing a key role in the development of early life forms and the evolution of metabolic pathways. Among peptides, those with enzyme-like activities occupy a unique position between peptides and enzymes, combining both structural flexibility and catalytic functionality. However, their full potential remains largely untapped. Further exploration of these enzyme-like peptides at the nanoscale could provide valuable insights into modern nanotechnology, biomedicine, and even the origins of life. Hence, this review introduces the groundbreaking concept of “peptide nanozymes (PepNzymes)”, which includes single peptides exhibiting enzyme-like activities, peptide-based nanostructures with enzyme-like activities, and peptide-based nanozymes, thus enabling the investigation of biological phenomena at nanoscale dimensions. Through the rational design of enzyme-like peptides or their assembly with nanostructures and nanozymes, researchers have found or created PepNzymes capable of catalyzing a wide range of reactions. By scrutinizing the interactions between the structures and enzyme-like activities of PepNzymes, we have gained valuable insights into the underlying mechanisms governing enzyme-like activities. Generally, PepNzymes play a crucial role in biological processes by facilitating small-scale enzyme-like reactions, speeding up molecular oxidation-reduction, cleavage, and synthesis reactions, leveraging the functional properties of peptides, and creating a stable microenvironment, among other functions. These discoveries make PepNzymes useful for diagnostics, cellular imaging, antimicrobial therapy, tissue engineering, anti-tumor treatments, and more while pointing out opportunities. Overall, this research provides a significant journey of PepNzymes’ potential in various biomedical applications, pushing them towards new advancements.

Abstract In foundation pit excavation, granite residual soil is frequently used as a foundation-bearing layer, necessitating the consideration of its unique engineering characteristics in stability studies. Numerical analysis is a crucial method for evaluating the stability of excavation pits, with the Modified Mohr–Coulomb (MMC) model showing good applicability. However, research on the MMC model in granite residual soil regions remains limited. This study investigates the impact of key stiffness parameters ( $${E}_{ur}^{ref}$$ E ur ref , $${E}_{oed}^{ref}$$ E oed ref , and $${E}_{50}^{ref}$$ E 50 ref ) of the MMC model on the stability of excavation pit retaining structures, using case studies from the Zhuhai area. The research addresses the selection of stiffness parameters for different soil layers and proposes a proportional relationship between model stiffness parameters and field test mechanical parameters. Using the horizontal displacement of the underground diaphragm wall as an indicator, single-factor sensitivity analysis and orthogonal experimental methods were employed to determine the sensitivity and proportional relationships of stiffness parameters in the Zhuhai region. The findings indicate that $${E}_{ur}^{ref}$$ E ur ref has the most significant impact on model uncertainty, while $${E}_{50}^{ref}$$ E 50 ref has the least effect on horizontal displacement. The ratios of $${E}_{ur}^{ref}$$ E ur ref / $${E}_{50}^{ref}$$ E 50 ref , $${E}_{oed}^{ref}$$ E oed ref / $${E}_{50}^{ref}$$ E 50 ref , and $${E}_{50}^{ref}$$ E 50 ref to the deformation modulus ( $${E}_{0}$$ E 0 ) provide essential references for numerical simulation of excavation pits in the Zhuhai area.

Abstract Background This study focused on the aging mechanism and degradation of mechanical and structural features of elastodontic appliances (EA) under artificial and intraoral aging to achieve oral myofunctional therapy with particular removable silicone elastomer devices. Materials and methods EAs artificially aged in saliva with different pH values were investigated through cyclic compression testing along with characterization techniques (Scanning electron microscopy, X-ray diffraction, and Fourier transform infrared spectroscopy), and characterization analysis was also performed on clinically retrieved EAs. Results Artificial aging was found to have minimal effect on the structural properties of EAs, and intraorally aged samples showed perceptible micro-morphology. The Mullins index and peak stress decreased (P

Abstract Background An adaptive design allows modifying the design based on accumulated data while maintaining trial validity and integrity. The final sample size may be unknown when designing an adaptive trial. It is therefore important to consider what sample size is used in the planning of the study and how that is communicated to add transparency to the understanding of the trial design and facilitate robust planning. In this paper, we reviewed trial protocols and grant applications on the sample size reporting for randomised adaptive trials. Method We searched protocols of randomised trials with comparative objectives on ClinicalTrials.gov (01/01/2010 to 31/12/2022). Contemporary eligible grant applications accessed from UK publicly funded researchers were also included. Suitable records of adaptive designs were reviewed, and key information was extracted and descriptively analysed. Results We identified 439 records, and 265 trials were eligible. Of these, 164 (61.9%) and 101 (38.1%) were sponsored by industry and public sectors, respectively, with 169 (63.8%) of all trials using a group sequential design although trial adaptations used were diverse. The maximum and minimum sample sizes were the most reported or directly inferred (n = 199, 75.1%). The sample size assuming no adaptation would be triggered was usually set as the estimated target sample size in the protocol. However, of the 152 completed trials, 15 (9.9%) and 33 (21.7%) had their sample size increased or reduced triggered by trial adaptations, respectively. The sample size calculation process was generally well reported in most cases (n = 216, 81.5%); however, the justification for the sample size calculation parameters was missing in 116 (43.8%) trials. Less than half gave sufficient information on the study design operating characteristics (n = 119, 44.9%). Conclusion Although the reporting of sample sizes varied, the maximum and minimum sample sizes were usually reported. Most of the trials were planned for estimated enrolment assuming no adaptation would be triggered. This is despite the fact a third of reported trials changed their sample size. The sample size calculation was generally well reported, but the justification of sample size calculation parameters and the reporting of the statistical behaviour of the adaptive design could still be improved.

BackgroundEpstein–Barr virus (EBV) is a significant global public health concern because of its association with various malignancies and autoimmune diseases. Over 90% of the global population is chronically infected with EBV, impacting numerous cancer-related cases annually. However, none of the effective prophylactic vaccines against EBV is approved at present.MethodsIn this study, we developed a novel vaccine candidate based on epitope peptides from the receptor-binding domain of EBV-encoded gp350 glycoprotein to prevent EBV infection. These epitope peptides detected a binding capability with host cells were then fused by flexibility linkers and expressed in Escherichia coli to reduce the unnecessary glycan modifications to simulate their free-glycan status. The fused recombinant protein (L350) was displayed on the surface of ferritin-based nanoparticle. The immunogenicity of the L350–ferritin nanoparticle was evaluated in Balb/c mice, and the neutralizing titers of sera from immunized mice were detected by means of an infection blocking assay in an in vitro cell model.ResultsAll the five epitope peptides could bind to AKATA cells, and their fused recombinant protein (L350) was successfully presented on the surface of self-assembled ferritin nanoparticles. Sera from the L350–ferritin nanoparticle-immunized mice showed high titers of both L350 protein-specific and gp350D123 protein-specific antibodies, and sera from gp350D123 protein-immunized mice could also recognize L350 protein well. Most importantly, the L350–ferritin nanoparticle induced efficient neutralizing antibodies to block EBV-GFP infection in AKATA cells and also constructed a strong antigen-specific B-cell memory in immunized mice. Moreover, histopathological changes of main tissues from all vaccinated mice were not observed.ConclusionThese data indicate that the L350–ferritin nanoparticle vaccine candidate has considerable potential application in preventing EBV infection and provides a promising basis for developing prophylactic EBV vaccines.

ABSTRACT Background As the primary organ of metabolism and detoxification, the liver may contribute to the pathogenesis of lung cancer. We aimed to illuminate the intricate link between liver function biomarkers and lung cancer risk, as well as delineate the role of smoking behavior within this association. Methods We investigated the associations of seven liver function biomarkers levels (alkaline phosphatase [ALP], alanine transaminase [ALT], total bilirubin [TBIL], albumin [ALB], gamma‐glutamyltransferase [GGT], aspartate transaminase [AST], and total protein [TP]) with lung cancer risk across the UK Biobank (N = 337 499) through restricted cubic splines and Cox proportional hazards models. Moreover, Mendelian randomization (MR) was utilized to evaluate the causal effect of smoking behavior on these biomarkers. Then a lung cancer risk prediction model was developed among smokers by backward stepwise logistic regression. Results During a median follow‐up of 13.3 years, 3003 lung cancer cases were identified. We found ALP levels positively associated with lung cancer risk, whereas ALT, TBIL, ALB, and AST were inversely correlated; TP exhibited a U‐shaped association, whereas GGT displayed a mirrored J‐shaped relationship. These associations were amplified among smokers. MR analysis indicated that smoking behavior could increase ALP (odds ratio [OR]: 1.05) and GGT (OR: 1.15) levels while decreasing TBIL (OR: 0.92), ALB (OR: 0.92), and TP (OR: 0.96) levels. The lung cancer risk model incorporating these biomarkers in smokers demonstrated robust discrimination. Conclusion Our finding provides perspectives and evidences towards the intricate crosstalk between the hepatic and pulmonary systems, as well as the processes through which tobacco catalyzes lung carcinogenesis.

The successful management of lepidopteran moths in orchards usually depends on the precise forecast of adult activity. However, the seasonal phenology of moths varies between crop cultivars and years, making it difficult to schedule the control measures. Here, we monitored male flight activity of oriental fruit moth Grapholita molesta and summer fruit tortrix moth Adoxophyes orana by using sex pheromone traps in peach orchards of three different cultivars for three successive years. We developed a logistic multiple-peaks model to fit data and then calculated degree-days (DD) required for male activity and neonate emergency. Results show that G. molesta and A. orana males had 4–5 and 3 flight peaks per year, respectively. The seasonal phenology of G. molesta or A. orana was quite stable with an identical timing of each flight peak between cultivars in a year. The flight activity was usually higher in the second and third peaks for both moths, with a higher cumulative number of G. molesta males captured than that of A. orana. Compared to A. orana, G. molesta emerged early in spring and required lower degree-days to reach the subsequent flight peaks and for neonate emergency. Our results suggest that to decline the possibility of outbreaks of moths during the growing seasons, pheromone traps should be scheduled in April with a cumulative DD between 49.6 and 207.1 for G. molesta and in mid-May–early June with a cumulative DD between 450.4 and 866.7 for A. orana, aiming to trap the newly emerged male adults or disrupting female mating success of overwintered moths in orchards. Based on the thermal requirement for egg hatching (i.e., 79.4 DD for G. molesta and 90.0 DD for A. orana), insecticide treatments would be applied in late-April–early May and late May–early June to reduce the field population density of neonates of G. molesta and A. orana, respectively, to reduce fruit damage in orchards. Furthermore, pheromone traps set up in late July–early August (573.8–1025.2 DD) for G. molesta and in mid-September (1539.7–1788.9 DD) for A. orana may suppress overwintering populations and thus decrease pest infestation in next year.

Abstract Background As a biennial plant, Secale cereale L is usually harvested in the autumn in the northern part of China where the temperature difference between day and night is of great disparity Through the pot experiment, the seedlings were cut to 2, 6 and 10 cm stubble height, and the simulated freeze-thaw (FT) stress (10/− 5 °C) was carried out after 6 days regrowth. The physiological effects of FT with different stubble height were revealed by analyzing the relative water content (RWC), osmotic adjustment substance concentration (soluble sugar and protein), membrane peroxidation (MDA) and catalase (CAT) activity. Results The results demonstrated that under freeze stress (− 5 °C), the content of soluble protein and MDA decreased and the seedlings of 2 cm treatment kept higher level of soluble protein and MDA, while the seedlings of 6 and 10 cm treatments kept higher level of the RWC, soluble sugar content, and CAT activity. After FT stress, the content of soluble sugar and protein, RWC in the 6 cm treatment were higher than those in 2 cm and 10 cm treatments, and the CAT activity in 10 cm treatment was the highest while the MDA content is lower. Conclusion These data suggest that keeping high stubble height is more adaptive for short-term FT stress.

Abstract Chimeric antigen receptor-based T-cell immunotherapy is a promising strategy for treatment of hematological malignant tumors; however, its efficacy towards solid cancer remains challenging. We therefore focused on developing nanobody-based CAR-T cells that treat the solid tumor. CD105 expression is upregulated on neoangiogenic endothelial and cancer cells. CD105 has been developed as a drug target. Here we show the generation of a CD105-specific nanobody, an anti-human CD105 CAR-T cells, by inserting the sequences for anti-CD105 nanobody-linked standard cassette genes into AAVS1 site using CRISPR/Cas9 technology. Co-culture with CD105+ target cells led to the activation of anti-CD105 CAR-T cells that displayed the typically activated cytotoxic T-cell characters, ability to proliferate, the production of pro-inflammatory cytokines, and the specific killing efficacy against CD105+ target cells in vitro. The in vivo treatment with anti-CD105 CAR-T cells significantly inhibited the growth of implanted CD105+ tumors, reduced tumor weight, and prolonged the survival time of tumor-bearing NOD/SCID mice. Nanobody-based CAR-T cells can therefore function as an antitumor agent in human tumor xenograft models. Our findings determined that the strategy of nanobody-based CAR-T cells engineered by CRISPR/Cas9 system has a certain potential to treat solid tumor through targeting CD105 antigen.

Abstract Background This study aims to assess the association between body mass index (BMI) and health-related quality of life (HRQOL), and to further explore gender differences in BMI-HRQOL association among adults. Methods We used data from the fifth Health Service Survey of Shandong Province, which was part of China’s National Health Service Survey (NHSS), a total of 27,257 adults aged 18 and over were interviewed. The HRQOL was measured using the EuroQOL-5 Dimensions (EQ-5D) instrument. One-way ANOVA and Post hoc tests were used to compare EQ-5D utility values and visual analogue scale (VAS) scores between BMI categories. Tobit regression models were used to identify the association between BMI and HRQOL for male and female separately after controlling for influential confounders, and to assess gender differences on the relationship between BMI and HRQOL. Results The prevalence of underweight in men and women were 3.2 and 5.3%, respectively, while the prevalence of overweight/obesity in men and women were 35.7 and 34.6%, respectively. Men had higher EQ-5D utility values and VAS scores than women. The mean EQ-5D utility value and VAS score was highest in obese men and normal-weight women, respectively. After controlling potential confounders, being underweight was significantly and negatively associated with lower HRQOL among adults. The relationship between obesity and gender was that in women obesity was negatively and significantly associated with HRQOL, whereas in men this association was positive but not statistically significant. Results of gender by BMI interaction in regression model showed that this difference between men and women in this respect was significant. Conclusions The association between BMI and HRQOL differed by gender and the so-called “obesity-HRQOL paradox” phenomenon was verified in male adults. Gender difference should be considered when implementing targeted weight control programs and appropriate interventions to improve HRQOL.

Abstract Backgrounds Suicidal ideation is an important public health issue due to devastating mortality. In the meantime, interpersonal trust was found to be negatively associated with mental disorder and physical health. Although there is increasing evidence that interpersonal trust is a significant predictor of suicidal ideation, evidence of this association is still lacking in the developing world. The aim of this study was to test the association between interpersonal trust and suicidal ideation among older adults in China. Methods Using a multi-stage stratified sampling strategy, 7070 older adults aged 60 and above from Shandong Province, China were recruited in this study. Socio-demographic characteristics, health status, family relationship, psychological distress, interpersonal trust and suicidal ideation in the last 12 months were obtained through face to face interviews. The association between interpersonal trust and suicidal ideation was assessed using multiple logistic regression models adjusting for socio-demographic characteristics, health status, family relationship and psychological distress. Results 7.1% of participants reported suicidal ideation in the past 12 months, which was related to gender, resident area, marital status, educational level, self-rated economic, chronic disease, self-rated health status and family relationship within 1 month, psychological distress and interpersonal trust. After adjusting for sociodemographic factors, health status, family relationship and psychological distress, interpersonal mistrust was associated with two times odds of suicidal ideation when compared to interpersonal trust Conclusions The interpersonal trust was associated with suicidal ideation among elderly in Shandong, China. Intervention approaches regarding inducing and promoting interpersonal trust should be developed to prevent suicide.

Abstract Background Blood pressure measurement is the first step in preventing and controlling hypertension. The objective of this study is to examine the rural–urban difference towards blood pressure measurement among elderly with hypertension. Methods A total of 2007 elderly (65+) were selected from the fifth Health Service Survey of Shandong Province in 2013. A standardized questionnaire was used to investigate the demographic characters, socioeconomic status, self-rated health, and blood pressure related index. Three logistic regression models were used to examine the difference in blood pressure measurement between rural and urban elderly. Unadjusted and adjusted logistic regression models were used to explore the associated factors of blood pressure measurement in both rural areas and urban areas. Results The prevalence of weekly blood pressure measurement in urban elderly was higher than that in rural elderly (63.9% vs 34.3%). The rural elderly had an odds ratio (OR) for weekly blood pressure measurement of 0.467 (95%CI = 0.380–0.575) compared with urban elderly. Binary logistic regression analysis showed that medication frequency and accepting health care professionals’ guidance were common associated factors of blood pressure measurement among both rural and urban elderly; personal income was unique associated factor of blood pressure measurement among rural elderly; marital status, education level, self-rated health, and blood pressure level currently were unique associated factors of blood pressure measurement among urban elderly. Conclusions There is a big difference in blood pressure measurement between rural and urban elderly. Interventions targeting identified at-risk subgroups, especially for those rural elderly, should be made to reduce such a gap.

Abstract Protein therapeutics are essential in treating various diseases, but their inherent biological instability and short circulatory half-lives in vivo pose challenges. Herein, a quantitative one-pot iterative living polymerization technique is reported towards precision control over the molecular structure and monomer sequence of protein-polymer conjugates, aiming to maximize physicochemical properties and biological functions of proteins. Using this quantitative one-pot iterative living polymerization technique, we successfully develop a series of sequence-controlled protein-multiblock polymer conjugates, enhancing their biostability, pharmacokinetics, cellular uptake, and in vivo biodistribution. All-atom molecular dynamics simulations are performed to disclose the definite sequence-function relationship of the bioconjugates, further demonstrating their sequence-encoded cellular uptake behavior and in vivo biodistribution in mice. Overall, this work provides a robust approach for creating precision protein-polymer conjugates with defined sequences and advanced functions as a promising candidate in disease treatment.

The photo-response non-uniformity (PRNU) noise of imaging sensors significantly aids digital forensics and judicial identification, as it can be used as the fingerprint for uniquely identifying individual imaging devices. During the PRNU fingerprint extraction, it is very important for source camera identification to estimate the natural noise from real-world images. Most existing deep learning-based neural networks have a large number of model parameters, and they may not be practical in realistic scenarios such as deploying the model on small devices like smartphones and remote forensics equipment. In this paper, we present a lightweight PRNU fingerprint extraction algorithm based on an invertible denoising network (InvDN) for source camera identification. Specifically, to reduce the number of parameters, the deep network uses a constant amount of memory to compute the gradient and employs the same parameters for both forward and backward propagation. In addition, by introducing an information-loss-less reversible network, more complete PRNU fingerprint information can be extracted. Experimental results show that this algorithm exhibits superior PRNU fingerprint extraction performance and generalization ability compared to the state-of-the-art PRNU fingerprint extraction algorithms.

Triptolide (TP) is the major bioactive component of traditional Chinese medicine Tripterygium wilfordii Hook. F., a traditional Chinese medicinal plant categorized within the Tripterygium genus of the Celastraceae family. It is recognized for its therapeutic potential in addressing a multitude of diseases. Nonetheless, TP is known to exhibit multi-organ toxicity, notably hepatotoxicity, which poses a significant concern for the well-being of patients undergoing treatment. The precise mechanisms responsible for TP-induced hepatotoxicity remain unresolved. In our previous investigation, it was determined that TP induces heightened hepatic responsiveness to exogenous lipopolysaccharide (LPS). Additionally, natural killer (NK) cells were identified as a crucial effector responsible for mediating hepatocellular damage in this context. However, associated activating receptors and the underlying mechanisms governing NK cell represented innate lymphoid cell (ILC) activation remained subjects of inquiry and were not yet investigated. Herein, activating receptor Killer cell lectin like receptor K1 (NKG2D) of group 1 ILCs was specifically upregulated in TP- and LPS-induced acute liver failure (ALF), and in vivo blockade of NKG2D significantly reduced group 1 ILC mediated cytotoxicity and mitigated TP- and LPS-induced ALF. NKG2D ligand UL16-binding protein-like transcript 1 (MULT-1) was found upregulated in liver resident macrophages (LRMs) after TP administration, and LRMs did exhibit NK cell activating effect. Furthermore, M1 polarization of LRMs cells was observed, along with an elevation in intracellular tumor necrosis factor (TNF)-α levels. In vivo neutralization of TNF-α significantly alleviated TP- and LPS-induced ALF. In conclusion, the collaborative role of group 1 ILCs and LRMs in mediating hepatotoxicity was confirmed in TP- and LPS-induced ALF. TP-induced MULT-1 expression in LRMs was the crucial mechanism in the activation of group 1 ILCs via MULT-1-NKG2D signal upon LPS stimulation, emphasizing the importance of infection control after TP administration.

The aim of this study was to determine the optimal velocity loss (VL) threshold that maximises the post activation potentiation (PAP) stimulus for achieving larger and more consistent performance gains in track and field athletes. Twenty-two athletes from athletics participated in four back squat PAP tests with four different VL threshold (5%, 10%, 15% and 20% VL) at an intensity of 85%1RM. Countermovement jump (CMJ) height, power, and momentum were assessed before, and 10 s, 4, 8, 12, 16 minutes after the PAP condition. Repetitions of the squat in all the PAP conditions were also recorded. Only the 5% VL condition produced significant improvements in height (ES = 0.73, P = 0.038), peak power output (ES = 0.73, P = 0.038) and momentum (ES = 0.72, P = 0.041) of CMJ, and these changes appeared 8 minutes after the condition. The total number of repetitions during the 5% VL condition was significantly lower than that observed in the 15% (P = 0.003) and 20% VL (P < 0.001) trials. The results from this study indicate that 5%VL during the 2 sets preconditioning squat at 85%1RM was optimal for eliciting PAP in a CMJ exercise, and resulted in significant increases at the 8-min recovery period. The same squat condition also had the least number of repetitions. However, considering the efficiency in practice, athletes can also choose the rest time of 4-min, which can also achieve similar results.

Short-circuit fault and local demagnetization faults are two common fault types of permanent-magnet machines, aiming to improve the reliability of multi-phase PM machines in safety-critical applications, a method to distinguish these two kinds of faults in six-phase interior PM machines is proposed. The voltage characteristics in synchronous frame of local demagnetization fault and short-circuit fault are investigated, and the influences of load current and internal power factor angle on the voltage harmonic characteristics are analyzed. To solve the problem of the interaction between the extracted harmonic amplitudes in integer and non-integer orders, the average and variance of the harmonic coefficients of the voltages in synchronous frame are utilized to distinguish two types of faults. Through comparing the fault indices under different load conditions of the six-phase interior PM machine, the distinguishing ability of the method is validated.

Tripterygium wilfordii multiglycoside (GTW), extracted and purified from the peeled roots of T. wilfordii Hook.f. (TwHF), is a well-known traditional Chinese medicine and applied to various autoimmune diseases clinically. However, it has been reported to cause severe liver injury. At present, the mechanism underlying GTW-induced hepatotoxicity remain poorly defined. Here, we evaluated the effects of GTW on mouse liver and elucidated the associated mechanisms via label-free proteomics combined with bioinformatics analysis. Male C57BL/6J mice were randomly divided into normal group, a low-dose GTW (70 mg/kg) group and a high-dose GTW (140 mg/kg) group. After 1-week administration, GTW dose-dependently induced hepatotoxicity. Further analysis showed that GTW could act on the intestinal immune network for IgA production pathway, which plays an important role in maintaining intestinal homeostasis and influences the crosstalk between gut and liver. Western blots confirmed that GTW could decrease pIgR protein expression in the liver and ileum, and, as a result, the secretion of IgA into gut lumen was reduced. Further validation showed that intestinal barrier integrity was impaired in GTW-treated mice, promoting bacteria transferring to the liver and triggering proinflammatory response. Our study demonstrated that gut-liver axis may play a vital part in the progression of GTW-induced hepatotoxicity, which provides guidance for basic research and clinical application of GTW.

Cholestasis is a common, chronic liver disease that may cause fibrosis and cirrhosis. Tripterygium wilfordii Hook.f (TWHF) is a species in the Euonymus family that is commonly used as a source of medicine and food in Eastern and Southern China. Triptolide (TP) is an epoxy diterpene lactone of TWHF, as well as the main active ingredient in TWHF. Here, we used a mouse model of common bile duct ligation (BDL) cholestasis, along with cultured human intrahepatic biliary epithelial cells, to explore whether TP can relieve cholestasis. Compared with the control treatment, TP at a dose of 70 or 140 μg/kg reduced the serum levels of the liver enzymes alanine transaminase, aspartate aminotransferase, and alkaline phosphatase in mice; hematoxylin and eosin staining also showed that TP reduced necrosis in tissues. Both in vitro and in vivo analyses revealed that TP inhibited cholangiocyte proliferation by reducing the expression of RelB. Immunohistochemical staining of CK19 and Ki67, as well as measurement of Ck19 mRNA levels in hepatic tissue, revealed that TP inhibited the BDL-induced ductular reaction. Masson 3 and Sirius Red staining for hepatic hydroxyproline showed that TP alleviated BDL-induced hepatic fibrosis. Additionally, TP substantially inhibited BDL-induced hepatic inflammation. In summary, TP inhibited the BDL-induced ductular reaction by reducing the expression of RelB in cholangiocytes, thereby alleviating liver injury, fibrosis, and inflammation.

Previously, we proposed a new perspective of triptolide (TP)-associated hepatotoxicity: liver hypersensitivity upon lipopolysaccharide (LPS) stimulation. However, the mechanisms for TP/LPS-induced hepatotoxicity remained elusive. The present study aimed to clarify the role of LPS in TP/LPS-induced hepatotoxicity and the mechanism by which TP induces liver hypersensitivity upon LPS stimulation. TNF-α inhibitor, etanercept, was injected intraperitoneally into mice to investigate whether induction of TNF-α by LPS participated in the liver injury induced by TP/LPS co-treatment. Mice and hepatocytes pretreated with TP were stimulated with recombinant TNF-α to assess the function of TNF-α in TP/LPS co-treatment. Additionally, time-dependent NF-κB activation and NF-κB-mediated pro-survival signals were measured in vivo and in vitro. Finally, overexpression of cellular FLICE-inhibitory protein (FLIP), the most potent NF-κB-mediated pro-survival protein, was measured in vivo and in vitro to assess its function in TP/LPS-induced hepatotoxicity. Etanercept counteracted the toxic reactions induced by TP/LPS. TP-treatment sensitized mice and hepatocytes to TNF-α, revealing the role of TNF-α in TP/LPS-induced hepatotoxicity. Mechanistic studies revealed that TP inhibited NF-κB dependent pro-survival signals, especially FLIP, induced by LPS/TNF-α. Moreover, overexpression of FLIP alleviated TP/LPS-induced hepatotoxicity in vivo and TP/TNF-α-induced apoptosis in vitro. Mice and hepatocytes treated with TP were sensitive to TNF-α, which was released from LPS-stimulated immune cells. These and other results show that the TP-induced inhibition of NF-κB-dependent transcriptional activity and FLIP production are responsible for liver hypersensitivity.

Intrahepatic cholestasis is a kind of clinical syndrome along with hepatotoxicity which caused by intrahepatic and systemic accumulations of bile acid. There are several crucial generating factors of the pathogenesis of cholestasis, such as inflammation, dysregulation of bile acid transporters and oxidative stress. SIRT1 is regarded as a class III histone deacetylase (HDAC). According to a set of researches, SIRT1 is one of the most important factors which can regulate the hepatic bile acid metabolism. SRT1720 is a kind of activator of SIRT1 which is 1000 times more potent than resveratrol, and this paper is aimed to study its protective influence on hepatotoxicity and cholestasis induced by alpha-naphthylisothiocyanate (ANIT) in mice. The findings revealed that SRT1720 treatment increased FXR and Nrf2 gene expressions to shield against hepatotoxicity and cholestasis induced by ANIT. The mRNA levels of hepatic bile acid transporters were also altered by SRT1720. Furthermore, SRT1720 enhanced the antioxidative system by increasing Nrf2, SOD, GCLc, GCLm, Nqo1, and HO-1 gene expressions. In conclusion, a protective influence could be provided by SRT1720 to cure ANIT-induced hepatotoxicity and cholestasis, which was partly through FXR and Nrf2 activations. These results indicated that SIRT1 could be regarded as a therapeutic target to cure the cholestasis.