Host specificity of plant-associated bacteria is negatively associated with genome size and host abundance along a latitudinal gradient.

Host specialization plays a critical role in the ecology and evolution of plant-microbe symbiosis. Theory predicts that host specialization is associated with microbial genome streamlining and is influenced by the abundance of host species, both of which can vary across latitudes, leading to a latitudinal gradient in host specificity. Here, we quantified the host specificity and composition of plant-bacteria symbioses on leaves across 329 tree species spanning a latitudinal gradient. Our analysis revealed a predominance of host-specialized leaf bacteria. The degree of host specificity was negatively correlated with bacterial genome size and the local abundance of host plants. Additionally, we found an increased host specificity at lower latitudes, aligning with the high prevalence of small bacterial genomes and rare host species in the tropics. These findings underscore the importance of genome streamlining and host abundance in the evolution of host specificity in plant-associated bacteria along the latitudinal gradient.

In Operando Monitoring the Stress Evolution of Silicon Anode Electrodes during Battery Operation via Optical Fiber Sensors.

Silicon (Si) anode has attracted broad attention because of its high theoretical specific capacity and low working potential. However, the severe volumetric changes of Si particles during the lithiation process cause expansion and contraction of the electrodes, which induces a repeatedly repair of solid electrolyte interphase, resulting in an excessive consuming of electrolyte and rapid capacity decay. Clearly known the deformation and stress changing at µÎµ resolution in the Si-based electrode during battery operation provides invaluable information for the battery research and development. Here, an in operando approach is developed to monitor the stress evolution of Si anode electrodes via optical fiber Bragg grating (FBG) sensors. By implanting FBG sensor at specific locations in the pouch cells with different Si anodes, the stress evolution of Si electrodes has been systematically investigated, and Δσ/areal capacity is proposed for stress assessment. The results indicate that the differences in stress evolution are nested in the morphological changes of Si particles and the evolution characteristics of electrode structures. The proposed technique provides a brand-new view for understanding the electrochemical mechanics of Si electrodes during battery operation.

Rheumatoid arthritis patients harbour aberrant enteric bacteriophages with autoimmunity-provoking potential: a paired sibling study.

OBJECTIVES: Viruses have been considered as important participants in the development of rheumatoid arthritis (RA). However, the profile of enteric virome and its role in RA remains elusive. This study aimed to investigate the atlas and involvement of virome in RA pathogenesis. METHODS: Faecal samples from 30 pairs of RA and healthy siblings that minimise genetic interferences were collected for metagenomic sequencing. The α and ß diversity of the virome and the virome-bacteriome interaction were analysed. The differential bacteriophages were identified, and their correlations with clinical and immunological features of RA were analysed. The potential involvement of these differential bacteriophages in RA pathogenesis was further investigated by auxiliary metabolic gene annotation and molecular mimicry study. The responses of CD4+ T cells and B cells to the mimotopes derived from the differential bacteriophages were systemically studied. RESULTS: The composition of the enteric bacteriophageome was distorted in RA. The differentially presented bacteriophages correlated with the immunological features of RA, including anti-CCP autoantibody and HLA-DR shared epitope. Intriguingly, the glycerolipid and purine metabolic genes were highly active in the bacteriophages from RA. Moreover, peptides of RA-enriched phages, in particular Prevotella phage and Oscillibacter phage could provoke the autoimmune responses in CD4+ T cells and plasma cells via molecular mimicry of the disease-associated autoantigen epitopes, especially those of Bip. CONCLUSIONS: This study provides new insights into enteric bacteriophageome in RA development. In particular, the aberrant bacteriophages demonstrated autoimmunity-provoking potential that would promote the occurrence of the disease.

A nomogram to predict ventricular thrombus in dilated cardiomyopathy patients.

BACKGROUND: VT (Ventricular Thrombus) is a serious complication of dilated cardiomyopathy (DCM). Our goal is to develop a nomogram for personalized prediction of incident VT in DCM patients. METHODS: 1267 patients (52.87 ± 11.75 years old, 73.8% male) were analyzed retrospectively from January 01, 2015, to December 31, 2020. A nomogram model for VT risk assessment was established using minimum absolute contraction and selection operator (LASSO) and multivariate logistic regression analysis, and its effectiveness was validated by internal guidance. The model was evaluated by the area under the receiver operating characteristic curve (AUC), calibration curves, and decision curve analysis (DCA). We compared the performance in predicting VT between nomogram and CHA2DS2, CHA2DS2- VASc or ATRIA by AUC, akaike information criterion (AIC), bayesian information criterion (BIC), net reclassification index (NRI), and integrated discrimination index (IDI). RESULTS: 89 patients (7.02%) experienced VT. Multivariate logistic regression analysis revealed that age, left ventricular ejection fraction (LVEF), uric acid (UA), N-terminal precursor B-type diuretic peptide (NT-proBNP), and D-dimer (DD) were important independent predictors of VT. The nomogram model correctly separates patients with and without VT, with an optimistic C score of 0.92 (95%CI: 0.90-0.94) and good calibration (Hosmer-Lemeshow χ2 = 11.51, P = 0.12). Our model showed improved prediction of VT compared to CHA2DS2, CHA2DS2-VASc or ATRIA (all P < 0.05). CONCLUSIONS: The novel nomogram demonstrated better than presenting scores and showed an improvement in predicting VT in DCM patients.

Cross-scale spatial variability and associations of carbon pools provide insight into regulating carbon sequestration in tropical montane rainforests.

The spatial distribution of plant, soil, and microbial carbon pools, along with their intricate interactions, presents a great challenge for the current carbon cycle research. However, it is not clear what are the characteristics of the spatial variability of these carbon pools, particularly their cross-scale relationships. We investigated the cross-scale spatial variability of microbial necromass carbon (MNC), soil organic carbon (SOC) and plant biomass (PB), as well as their correlation in a tropical montane rainforest using multifractal analysis. The results showed multifractal spatial variations of MNC, SOC, and PB, demonstrating their adherence to power-law scaling. MNC, especially low MNC, exhibited stronger spatial heterogeneity and weaker evenness compared with SOC and PB. The cross-scale correlation between MNC and SOC was stronger than their correlations at the measurement scale. Furthermore, the cross-scale spatial variability of MNC and SOC exhibited stronger and more stable correlations than those with PB. Additionally, this research suggests that when SOC and PB are both low, it is advisable for reforestations to potentiate MNC formation, whereas when both SOC and PB are high some thinning can be advisable to favour MNC formation. Thus, these results support the utilization of management measures such as reforestation or thinning as nature-based solutions to regulate carbon sequestration capacity of tropical forests by affecting the correlations among various carbon pools.

Immobilized microalgae: principles, processes and its applications in wastewater treatment.

Microalgae have emerged as potential candidates for biomass production and pollutant removal. However, expensive biomass harvesting, insufficient biomass productivity, and low energy intensity limit the large-scale production of microalgae. To break through these bottlenecks, a novel technology of immobilized microalgae culture coupled with wastewater treatment has received increasing attention in recent years. In this review, the characteristics of two immobilized microalgae culture technologies are first presented and then their mechanisms are discussed in terms of biofilm formation theories, including thermodynamic theory, Derjaguin-Landau-Verwei-Overbeek theory (DLVO) and its extended theory (xDLVO), as well as ionic cross-linking mechanisms in the process of microalgae encapsulated in alginate. The main factors (algal strains, carriers, and culture conditions) affecting the growth of microalgae are also discussed. It is also summarized that immobilized microalgae show considerable potential for nitrogen and phosphorus removal, heavy metal removal, pesticide and antibiotic removal in wastewater treatment. The role of bacteria in the cultivation of microalgae by immobilization techniques and their application in wastewater treatment are clarified. This is economically feasible and technically superior. The problems and challenges faced by immobilized microalgae are finally presented.

Novel Circovirus in Blood from Intravenous Drug Users, Yunnan, China.

We identified a novel circovirus (human-associated circovirus 2 [HuCV2]) from the blood of 2 intravenous drug users in China who were infected with HIV-1, hepatitis C virus, or both. HuCV2 is most closely related to porcine circovirus 3. Our findings underscore the risk for HuCV2 and other emerging viruses among this population.

Diversity and biogeography of plant phyllosphere bacteria are governed by latitude-dependent mechanisms.

Predicting and managing the structure and function of plant microbiomes requires quantitative understanding of community assembly and predictive models of spatial distributions at broad geographic scales. Here, we quantified the relative contribution of abiotic and biotic factors to the assembly of phyllosphere bacterial communities, and developed spatial distribution models for keystone bacterial taxa along a latitudinal gradient, by analyzing 16S rRNA gene sequences from 1453 leaf samples taken from 329 plant species in China. We demonstrated a latitudinal gradient in phyllosphere bacterial diversity and community composition, which was mostly explained by climate and host plant factors. We found that host-related factors were increasingly important in explaining bacterial assembly at higher latitudes while nonhost factors including abiotic environments, spatial proximity and plant neighbors were more important at lower latitudes. We further showed that local plant-bacteria associations were interconnected by hub bacteria taxa to form metacommunity-level networks, and the spatial distribution of these hub taxa was controlled by hosts and spatial factors with varying importance across latitudes. For the first time, we documented a latitude-dependent importance in the driving factors of phyllosphere bacteria assembly and distribution, serving as a baseline for predicting future changes in plant phyllosphere microbiomes under global change and human activities.

Sesquiterpenoids from the Resina Commiphora Promoting the Apoptotic Activity of PC-3 Prostate Cancer Cells.

Four new germacrane-type sesquiterpenes commiphoranes M1-M4 (1-4) together with eighteen sesquiterpenes were isolated from the Resina Commiphora. The structures and relative configurations of new substances were determined by using spectroscopic methods. Biological activity investigation revealed that nine compounds including 7, 9, 14, 16, (+)-17, (-)-17, 18, 19, and 20 could induce the apoptosis of prostate cancer originated PC-3 cells, through classic apoptosis signaling pathway, even using flow cytometry showed that the compound (+)-17 caused apoptosis of PC-3 cells more than 40 %, suggesting their potential therapeutic application in the development of novel drugs against prostate cancer.

Arbuscular mycorrhizal fungi improve growth and tolerance of Platycladus orientalis under lead stress.

Platycladus orientalis is a significant woody plant for phytoremediation in heavy metals contaminated soils. The growth and tolerance of host plants under the lead (Pb) stress were enhanced by arbuscular mycorrhizal fungi (AMF). To evaluate the adjustment by AMF on growth and activity of antioxidant system of P. orientalis under Pb stress. The two-factor pot experiment was conducted with three AM fungal treatments (noninoculated, Rhizophagus irregularis, and Funneliformis mosseae) and four Pb levels (0, 500, 1000, and 2000 mg kg-1). AMF increased dry weight, phosphorus uptake, root vitality, and total chlorophyll content of P. orientalis in spite of Pb stress. Compared with nonmycorrhizal treatments, mycorrhizal P. orientalis had lower H2O2 and malondialdehyde (MDA) contents under Pb stress. AMF increased Pb uptake in roots and decreased the Pb translating to the shoots yet under Pb stress. Total glutathione and ascorbate in roots of P. orientalis were decreased by AMF inoculation. Mycorrhizal P. orientalis had higher superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and glutathione S-transferase (GST) activities in shoots and roots than nonmycorrhizal counterparts. Mycorrhizal P. orientalis under Pb stress showed higher expression of PoGST1 and PoGST2 in roots than that in CK treatments. Future studies will explore the function of induced tolerance genes by AMF of P. orientalis under Pb stress.

Arbuscular mycorrhizal fungi (AMF) decreased the reduced toxicity of lead to Platycladus orientalis under lead stress, including improving growth, root activity, photosynthesis, and antioxidant system activity, while reducing its oxidative damage. At the same time, lead inhibited the symbiosis between AMF and Platycladus orientalis.

Experimental and numerical study on Zn and Pb migration in the farmland soil under wastewater irrigation conditions.

Long-term wastewater irrigation may lead to the accumulation, transformation, and migration of heavy metals in the farmland soil, increasing the risk of groundwater pollution. However, it is currently uncertain whether using wastewater for irrigation would lead to the migration of heavy metals zinc (Zn) and lead (Pb) into deeper layers of soil, in the local undeveloped wastewater irrigation farmland. In this study, the migration characteristics of Zn and Pb from irrigation wastewater in local farmland soil were investigated through a series of experiments including adsorption experiments, tracer, and heavy metals breakthrough experiments, as well as numerical simulations using HYDRUS-2D software. The results revealed that the Langmuir adsorption model, CDE model, and TSM model were effective in fitting the required adsorption and solute transport parameters for the simulations. Furthermore, both the soil experiments and simulation results showed that in the test soil, Pb had a stronger affinity for adsorption sites than Zn, while Zn exhibited greater mobility than Pb. After 10 years of wastewater irrigation, it was found that Zn had migrated to a maximum depth of 32.69 cm underground and Pb had only migrated to 19.59 cm. Despite their migration, the two heavy metals have not yet reached the groundwater zone. Instead, they had accumulated to higher concentrations in the local farmland soil. Moreover, the proportion of active forms of Zn and Pb decreased after flooded incubation. The present results can improve understanding of the environmental behavior of Zn and Pb in the farmland soil and can be used as a basis for risk assessment of Zn and Pb polluting groundwater.

Atomically Dispersed CoN3 C1 -TeN1 C3 Diatomic Sites Anchored in N-Doped Carbon as Efficient Bifunctional Catalyst for Synergistic Electrocatalytic Hydrogen Evolution and Oxygen Reduction.

A encapsulation-adsorption-pyrolysis strategy for the construction of atomically dispersed Co-Te diatomic sites (DASs) that are anchored in N-doped carbon is reported as an efficient bifunctional catalyst for electrocatalytic hydrogen evolution reaction (HER) and oxygen reduction reaction (ORR). The as-constructed catalyst shows the stable CoN3 C1 -TeN1 C3 coordination structure before and after HER and ORR. The *OOH/*H intermediate species are captured by in situ Raman and in situ attenuated total reflectance-surface enhanced infrared absorption spectroscopy, indicating that the reactant O2 /H2 O molecule has a strong interaction with the Co site, revealing that Coδ+ is an effective active site. Theoretical calculations show that the Coδ+ has adsorption-activation function and the neighboring Teδ+ acts as an electron donor adjusting the electronic structure of Coδ+ , promoting the dissociation of H2 O molecules and the adsorption of H and oxygen-containing intermediates in HER and ORR. In the meanwhile, the nearest C atom around Co also profoundly affects the adsorption of H atoms. This results in the weakening of the OH adsorption and enhancement of H adsorption, as well as the more stable water molecule dissociation transition state, thus significantly boosting ORR and HER performance.

Virome of Bat Guano from Nine Northern California Roosts.

Bats are hosts to a large variety of viruses, including many capable of cross-species transmissions to other mammals, including humans. We characterized the virome in guano from five common bat species in 9 Northern California roosts and from a pool of 5 individual bats. Genomes belonging to 14 viral families known to infect mammals and 17 viral families infecting insects or of unknown tropism were detected. Nearly complete or complete genomes of a novel parvovirus, astrovirus, nodavirus, circular Rep-encoding single-stranded DNA (CRESS-DNA) viruses, and densoviruses, and more partial genomes of a novel alphacoronavirus and a bunyavirus were characterized. Lower numbers of reads with >90% amino acid identity to previously described calicivirus, circovirus, adenoviruses, hepatovirus, bocaparvoviruses, and polyomavirus in other bat species were also found, likely reflecting their wide distribution among different bats. Unexpectedly, a few sequence reads of canine parvovirus 2 and the recently described mouse kidney parvovirus were also detected and their presence confirmed by PCR; these possibly originated from guano contamination by carnivores and rodents. The majority of eukaryotic viral reads were highly divergent, indicating that numerous viruses still remain to be characterized, even from such a heavily investigated order as Chiroptera.IMPORTANCE Characterizing the bat virome is important for understanding viral diversity and detecting viral spillover between animal species. Using an unbiased metagenomics method, we characterize the virome in guano collected from multiple roosts of common Northern California bat species. We describe several novel viral genomes and report the detection of viruses with close relatives reported in other bat species, likely reflecting cross-species transmissions. Viral sequences from well-known carnivore and rodent parvoviruses were also detected, whose presence are likely the result of contamination from defecation and urination atop guano and which reflect the close interaction of these mammals in the wild.

Multi-channel parallel ultrasound detection based on a photothermal tunable fiber optic sensor array.

A multi-channel parallel ultrasound detection system based on a photothermal tunable fiber optic sensor array is proposed. The resonant wavelength of the ultrasound sensor has a quadratic relationship with the power of a 980-nm heating laser. The maximum tuning range is larger than 15 nm. Through photothermal tuning, the inconsistent operating wavelengths of the Fabry-Perot (FP) sensor array can be solved, and then a multiplexing capacity of up to 53 can be theoretically realized, which could greatly reduce the time required for data acquisition. Then, a fixed wavelength laser with ultra-narrow linewidth is used to interrogate the sensor array. The interrogation system demonstrates a noise equivalent pressure (NEP) as low as 0.12 kPa, which is 5.5-times lower than the commercial hydrophone. Furthermore, a prototype of a four-channel ultrasound detection system is built to demonstrate the parallel detection capability. Compared with the independent detection, the SNR of parallel detection does not deteriorate, proving that the parallel detection system and the sensor array own very low cross talk characteristics. The parallel detection technique paves a way for real-time photoacoustic/ultrasound imaging.

Experimental and numerical study on Cu and Cd migration in different functional-area soils under simulated rainfall conditions.

Natural rainfall exerts a significant influence on the migration of heavy metals in soil. However, the knowledge of migration characteristics and release kinetics of heavy metals in contaminated soils under different rainfall intensities still remains unclear. In this study, the simulated rainfall of different intensities was designed to experimentally and numerically investigate Cu and Cd movements in different functional-area (agriculture, industrial, urban) soils. A HYDRUS-2D model was optimized to simulate the migration process of Cu and Cd in soil under different rainfall conditions. The hydraulic properties and solute transport parameters used in the model were estimated based on isothermal adsorption and chloride ion penetration experimental measurements and related model fitting. Furthermore, Cu and Cd BTCs (Breakthrough Curves) were fitted using the HYDRUS-2D inverse solution function with two-site model. The results showed that the order of the migration capacity of Cu and Cd in different functional-area soils was agriculture soil > industrial soil > urban soil, and Cd had a greater risk of groundwater pollution than Cu. With the increase of rainfall intensity, the high proportion of the exchangeable state of Cu and Cd in contaminated soil is easy to be released. Furthermore, the model was proved to describe the distribution of Cu and Cd in the soil profile very well. The present results can improve understanding of the environmental behavior of Cu and Cd in different functional-areas soils and can be used as a basis for risk assessment of Cu and Cd polluting groundwater.

Faecalibacter rhinopitheci sp. nov., a bacterium isolated from the faeces of Rhinopithecus bieti.

A novel Gram-stain-negative strain, WQ 117T, isolated from the faeces of Rhinopithecus bieti collected at Yunnan Snub-nosed Monkey National Park, Yunnan province, PR China, was subjected to a polyphasic taxonomic study. The results of phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolate represented a member of the genus Faecalibacter, sharing 97.64 % sequence similarity with the type strain Faecalibacter macacae YIM 102668T. The G+C content of the genomic DNA of WQ117T was 30.5 mol%. The predominant isoprenoid quinone was MK-6. The major cellular fatty acids was iso-C15 : 0. The whole genome average nucleotide identity (gANI) values and the digital DNA-DNA hybridization values between WQ 117T and YIM 102668T were 79.66 % and 22.20 %, respectively. Growth occurred at 0-50 °C (optimally at 28-35 °C), pH 7.0-9.0 (optimally at pH 8.0) and with 0-2 % (w/v) NaCl (optimally without NaCl). On the basis of the taxonomic evidence, a novel species, Faecalibacter rhinopitheci sp. nov., is proposed. The type strain is WQ 117T (=KCTC 82394T=CCTCC AA 2020027T).

Evolutionary dynamics of group A and B respiratory syncytial virus in China, 2009-2018.

Respiratory syncytial virus (RSV) is a major cause of acute respiratory tract infections in children and is a public health threat globally. To investigate the spatiotemporal dynamics of RSV evolution, we performed systematic phylogenetic analysis using all available sequences from the GenBank database, together with sequences from Shanghai, China. Both RSV-A and RSV-B appear to have originated in North America, with an inferred origin time of 1954.0 (1938.7-1967.6) and 1969.7 (1962.6-1975.5), respectively. BA-like strains of RSV-B, with a 60-nt insertion, and the ON1 strain of RSV-A, with a 72-nt insertion, emerged in 1997.6 (1996.2-1998.6) and 2010.1 (2009.1-2010.3), respectively. Since their origin, both genotypes have gradually replaced the former circulating genotypes to become the dominant strain. The population dynamic of RSV-A showed a seasonal epidemic pattern with obvious expansion in the periods of 2006-2007, 2010-2011, 2011-2012, and 2013-2014. Thirty fixed amino acid substitutions were identified during the divergence of NA4 from GA1 genotypes of RSV-A, and 13 were found during the divergence of SAB4 from GB1 of RSV-B. Importantly, ongoing evolution has occurred since the emergence of ON1, including four amino acid substitutions (I208L, E232G, T253K, and P314L). RSV-A genotypes GA5, NA4, NA1, and ON1 and RSV-B genotypes CB1, SAB4, BA-C, BA10, BA7, and BA9 were co-circulating in China from 2005 to 2015. In particular, RSV-A genotype ON1 was first detected in China in 2011, and it completely replaced GA2 to become the predominant strain after 2016. These data provide important insights into the evolution and epidemiology of RSV.

Gancochlearols E - I, meroterpenoids from Ganoderma cochlear against COX-2 and triple negative breast cancer cells and the absolute configuration assignment of ganomycin K.

Five new meroterpenoids, gancochlearols E - I (1, 3-6), and one compound ganomycin K (2) were isolated from the fruiting bodies of G. cochlear. Their structures were assigned by 1D and 2D NMR, MS, and CD analysis. Rh2(OCOCF3)4-induced ECD method was used to clarify the absolute configuration of secondary alcohol in 1 and 2. Biochemical evaluation showed that all the isolates significantly inhibit COX-2 enzyme in vitro with the IC50 values range from 1.03 µM to 2.71 µM. Further cellular assay revealed that (+)-3 and (-)-6 could suppress metastatic phenotype of triple-negative breast cancer (TNBC) cells via impeding the epithelial-mesenchymal transition (EMT).

In-situ DNA hybridization detection based on a reflective microfiber probe.

A label-free biosensor based on a reflective microfiber probe for in-situ real-time DNA hybridization detection is proposed and experimentally demonstrated. The microfiber probe is simply fabricated by snapping a non-adiabatic biconical microfiber through closing the oxyhydrogen flame during fiber stretching. Assisted with the Fresnel reflection at the end of microfiber, a reflective microfiber modal interferometer is realized. The in-situ DNA hybridization relies on the surface functionalization of a monolayer of Poly-L-lysine (PLL) and synthetic DNA sequences that bind to a given target with high specificity. The detection processes of DNA hybridization in various concentration of target DNA solutions are monitored in real-time and the experimental results present a minimum detectable concentration of 10pM with good repeatability. Additionally, the detection specificity is also investigated by immersing the microfiber probe into the non-complementary ssDNA solutions and observing the spectral variation. The proposed biosensor has advantages of high sensitivity, compact size, ease of use and simple fabrication, which makes it has great potential to be applied in a lot of fields such as disease diagnosis, medicine, and environmental science.

The p66shc-mediated Regulation of Hepatocyte Senescence Influences Hepatic Steatosis in Nonalcoholic Fatty Liver Disease.

BACKGROUND Recent studies have suggested that hepatocyte senescence could contribute to hepatic steatosis and its progression in nonalcoholic fatty liver disease (NAFLD). However, the underlying mechanism causing hepatocyte senescence in this pathological condition is still unclear. A thorough understanding of the mechanism could provide a new target for therapeutic intervention. The purpose of this study was to investigate the role of p66shc in hepatocyte senescence and hepatocyte damage in NAFLD progression. MATERIAL AND METHODS We examined the expression levels of hepatic p66shc and senescence markers in rats and humans with NAFLD, and we assessed the effect of p66shc knockdown or overexpression on senescence and steatosis in human liver cells. RESULTS In this study, we showed that increased hepatic p66shc expression was consistent with upregulated expression of the following senescence markers in NAFLD rats: heterochromatin protein-1-beta (HP1ß), p16, p21, and p53. Furthermore, senescence and steatosis could be induced in hepatoblastoma cell line (HepG2) cells when cells were stimulated with a low concentration of H2O2, and this effect was significantly alleviated by knockdown of p66shc. However, overexpression of p66shc could promote senescence and steatosis in L02 cells. Finally, increased hepatic p66shc protein levels correlated with enhanced expression of the senescence marker p21 and mirrored the degree of disease severity in NAFLD patients. CONCLUSIONS Our findings indicated that the increase in hepatocyte senescence and steatosis in NAFLD may be caused by the upregulation of p66shc expression, implying that strategies for p66shc-mediated regulation of hepatocyte senescence may provide new therapeutic tools for NAFLD.