CT Reconstruction using Diffusion Posterior Sampling conditioned on a Nonlinear Measurement Model.

Diffusion models have been demonstrated as powerful deep learning tools for image generation in CT reconstruction and restoration. Recently, diffusion posterior sampling, where a score-based diffusion prior is combined with a likelihood model, has been used to produce high quality CT images given low-quality measurements. This technique is attractive since it permits a one-time, unsupervised training of a CT prior; which can then be incorporated with an arbitrary data model. However, current methods rely on a linear model of x-ray CT physics to reconstruct or restore images. While it is common to linearize the transmission tomography reconstruction problem, this is an approximation to the true and inherently nonlinear forward model. We propose a new method that solves the inverse problem of nonlinear CT image reconstruction via diffusion posterior sampling. We implement a traditional unconditional diffusion model by training a prior score function estimator, and apply Bayes rule to combine this prior with a measurement likelihood score function derived from the nonlinear physical model to arrive at a posterior score function that can be used to sample the reverse-time diffusion process. This plug-and-play method allows incorporation of a diffusion-based prior with generalized nonlinear CT image reconstruction into multiple CT system designs with different forward models, without the need for any additional training. We develop the algorithm that performs this reconstruction, including an ordered-subsets variant for accelerated processing and demonstrate the technique in both fully sampled low dose data and sparse-view geometries using a single unsupervised training of the prior.

Exploring the mechanism of LncRNA CASC15 affecting hepatocellular carcinoma through miRNA.

This study aimed to determine the potential mechanisms through which long noncoding (Lnc) RNA cancer susceptibility candidate 15 (CASC15) affects hepatocellular carcinoma (HCC). We retrieved HCC RNA-seq and clinical information from the UCSC Xena database. The differential expression (DE) of CASC15 was detected. Overall survival was analyzed using Kaplan-Meier (K-M) curves. Molecular function and signaling pathways affected by CASC15 were determined using Gene Set Enrichment Analysis. Associations between CASC15 and the HCC microenvironment were investigated using immuno-infiltration assays. A differential CASC15-miRNA-mRNA network and HCC-specific CASC15-miRNA-mRNA ceRNA network were constructed. The overexpression of CASC15 in HCC tissues was associated with histological grade, clinical stage, pathological T stage, poor survival, more complex immune cell components, and 12 immune checkpoints. We identified 27 DE miRNAs and 270 DE mRNAs in the differential CASC15-miRNA-mRNA network, and 10 key genes that were enriched in 12 cancer-related signaling pathways. Extraction of the HCC-specific CASC15-miRNA-mRNA network revealed that IGF1R, MET, and KRAS were associated with HCC progression and occurrence. Our bioinformatic findings confirmed that CASC15 is a promising prognostic biomarker for HCC, and elevated levels in HCC are associated with the tumor microenvironment. We also constructed a disease-specific CASC15-miRNA-mRNA regulatory ceRNA network that provides a new perspective for the precise indexing of patients with elevated levels of CASC15.

A new method proposed for realizing human gait pattern recognition: Inspirations for the application of sports and clinical gait analysis.

BACKGROUND: Finding the best subset of gait features among biomechanical variables is considered very important because of its ability to identify relevant sports and clinical gait pattern differences to be explored under specific study conditions. This study proposes a new method of metaheuristic optimization-based selection of optimal gait features, and then investigates how much contribution the selected gait features can achieve in gait pattern recognition. METHODS: Firstly, 800 group gait datasets performed feature extraction to initially eliminate redundant variables. Then, the metaheuristic optimization algorithm model was performed to select the optimal gait feature, and four classification algorithm models were used to recognize the selected gait feature. Meanwhile, the accuracy results were compared with two widely used feature selection methods and previous studies to verify the validity of the new method. Finally, the final selected features were used to reconstruct the data waveform to interpret the biomechanical meaning of the gait feature. RESULTS: The new method finalized 10 optimal gait features (6 ankle-related and 4-related knee features) based on the extracted 36 gait features (85 % variable explanation) by feature extraction. The accuracy in gait pattern recognition among the optimal gait features selected by the new method (99.81 % ± 0.53 %) was significantly higher than that of the feature-based sorting of effect size (94.69 % ± 2.68 %), the sequential forward selection (95.59 % ± 2.38 %), and the results of previous study. The interval between reconstructed waveform-high and reconstructed waveform-low curves based on the selected feature was larger during the whole stance phase. SIGNIFICANCE: The selected gait feature based on the proposed new method (metaheuristic optimization-based selection) has a great contribution to gait pattern recognition. Sports and clinical gait pattern recognition can benefit from population-based metaheuristic optimization techniques. The metaheuristic optimization algorithms are expected to provide a practical and elegant solution for sports and clinical biomechanical feature selection with better economy and accuracy.

Polydatin inhibited TNF-α-induced apoptosis of skeletal muscle cells through AKT-mediated p38 MAPK and NF-κB pathways.

Skeletal muscle atrophy severely impacts one's quality of life. The effects and mechanism of polydatin on skeletal muscle atrophy are unclear. This study investigated the effects and mechanism of polydatin on TNF-α-induced skeletal muscle cells. The skeletal muscle cell atrophy model was established by inducing C2C12 cells with TNF-α. Cell viability, IL-1ß levels and cell apoptosis were assessed. The mRNA and protein expression levels of apoptosis-related proteins were measured. Meanwhile, the binding of polydatin to AKT was analyzed by molecular docking. TNF-α reduced cell fusion and viability while up-regulated IL-1ß level and promoted cell apoptosis. TNF-α activated AKT, NF-κB, and p38 MAPK signaling pathways. Polydatin reversed these effects induced by TNF-α, with a low concentration being more effective. Polydatin was predicted to bind to GLY162, PHE161, GLU198, THR195 and GLU191 sites of AKT protein through van der Waals force and conventional hydrogen bonds. Overexpression of AKT led to increased phosphorylation levels of AKT, p38, and p65 proteins, as well as IL-1ß levels and cell apoptosis. Polydatin inhibited TNF-α-induced apoptosis of C2C12 cells by regulating NF-κB and p38 MAPK signaling pathways through AKT. This suggests that polydatin shows promise as a new drug for the treatment of skeletal muscle atrophy.

Prognostic significance of LRRC1 in hepatocellular carcinoma and construction of relevant prognostic model.

This study aimed to elucidate the prognostic value of the leucine rich repeat containing 1 (LRRC1) gene in hepatocellular carcinoma (HCC) and to determine the effects of high and low LRRC1 expression on mutation and immune cell infiltration. We downloaded HCC mRNA-seq expression and clinical data from University of California Santa Cruz Xena. The expression of LRRC1 was compared between HCC tumor and normal samples. Tumor samples were divided according to high and low LRRC1 expression. Differentially expressed genes between the 2 groups were identified, and function, mutation, and immune cell infiltration were analyzed. Genes associated with immune cells were identified using weighted gene co-expression network analysis, and transcription factors of these genes were predicted. Moreover, a prognostic model was developed and its performance was evaluated. The expression of LRRC1 was upregulated in HCC tissues, and this indicated a poor prognosis for patients with HCC. Differentially expressed genes between high and low LRRC1 expression were significantly enriched in pathways associated with cancer, amino acid metabolism, carbohydrate metabolism, and the immune system. We identified 15 differentially infiltrated immune cells between tumors with high and low LRRC1 expression and 14 of them correlated with LRRC1 gene expression. Weighted gene co-expression network analysis identified 83 immune cell-related genes, 27 of which had prognostic value. Cyclic AMP-response element binding protein regulated annexin A5, matrix metallopeptidase 9, and LRRC1 in the transcription factor regulatory network. Finally, a prognostic model composed of 7 genes were generated, which could accurately predict the prognosis of HCC patients. The LRRC1 gene might serve as a potential immune-associated prognostic biomarker for HCC.

Identification of molecular subtypes and prognostic signatures based on transient receptor potential channel-related genes to predict the prognostic risk of hepatocellular carcinoma: A review.

Abnormal transient receptor potential (TRP) channel function interferes with intracellular calcium-based signaling and causes malignant phenotypes. However, the effects of TRP channel-related genes on hepatocellular carcinoma (HCC) remain unclear. This study aimed to identify HCC molecular subtypes and prognostic signatures based on TRP channel-related genes to predict prognostic risks. Unsupervised hierarchical clustering was applied to identify HCC molecular subtypes using the expression data of TRP channel-related genes. This was followed by a comparison of the clinical and immune microenvironment characteristics between the resulting subtypes. After screening for differentially expressed genes among subtypes, prognostic signatures were identified to construct risk score-based prognostic and nomogram models and predict HCC survival. Finally, tumor drug sensitivities were predicted and compared between the risk groups. Sixteen TRP channel-related genes that were differentially expressed between HCC and non-tumorous tissues were used to identify 2 subtypes. Cluster 1 had higher TRP scores, better survival status, and lower levels of clinical malignancy. Immune-related analyses also revealed higher infiltration of M1 macrophages and higher immune and stromal scores in Cluster 1 than in Cluster 2. After screening differentially expressed genes between subtypes, 6 prognostic signatures were identified to construct prognostic and nomogram models. The potential of these models to assess the prognostic risk of HCC was further validated. Furthermore, Cluster 1 was more distributed in the low-risk group, with higher drug sensitivities. Two HCC subtypes were identified, of which Cluster 1 was associated with a favorable prognosis. Prognostic signatures related to TRP channel genes and molecular subtypes can be used to predict HCC risk.

New Insights for the Design of Bionic Robots: Adaptive Motion Adjustment Strategies During Feline Landings.

Felines have significant advantages in terms of sports energy efficiency and flexibility compared with other animals, especially in terms of jumping and landing. The biomechanical characteristics of a feline (cat) landing from different heights can provide new insights into bionic robot design based on research results and the needs of bionic engineering. The purpose of this work was to investigate the adaptive motion adjustment strategy of the cat landing using a machine learning algorithm and finite element analysis (FEA). In a bionic robot, there are considerations in the design of the mechanical legs. (1) The coordination mechanism of each joint should be adjusted intelligently according to the force at the bottom of each mechanical leg. Specifically, with the increase in force at the bottom of the mechanical leg, the main joint bearing the impact load gradually shifts from the distal joint to the proximal joint; (2) the hardness of the materials located around the center of each joint of the bionic mechanical leg should be strengthened to increase service life; (3) the center of gravity of the robot should be lowered and the robot posture should be kept forward as far as possible to reduce machine wear and improve robot operational accuracy.

The Heterogeneous Severity of COVID-19 in African Countries: A Modeling Approach.

The COVID-19 pandemic has had a considerable impact on global health and economics. The impact in African countries has not been investigated thoroughly via fitting epidemic models to the reported COVID-19 deaths. We downloaded the data for the 12 most-affected countries with the highest cumulative COVID-19 deaths to estimate the time-varying basic reproductive number ([Formula: see text]) and infection attack rate. We develop a simple epidemic model and fitted it to reported COVID-19 deaths in 12 African countries using iterated filtering and allowing a flexible transmission rate. We observe high heterogeneity in the case-fatality rate across the countries, which may be due to different reporting or testing efforts. South Africa, Tunisia, and Libya were most affected, exhibiting a relatively higher [Formula: see text] and infection attack rate. Thus, to effectively control the spread of COVID-19 epidemics in Africa, there is a need to consider other mitigation strategies (such as improvements in socioeconomic well-being, healthcare systems, the water supply, and awareness campaigns).

The ubiquitin-dependent ATPase p97 removes cytotoxic trapped PARP1 from chromatin.

Poly (ADP-ribose) polymerase (PARP) inhibitors elicit antitumour activity in homologous recombination-defective cancers by trapping PARP1 in a chromatin-bound state. How cells process trapped PARP1 remains unclear. Using wild-type and a trapping-deficient PARP1 mutant combined with rapid immunoprecipitation mass spectrometry of endogenous proteins and Apex2 proximity labelling, we delineated mass spectrometry-based interactomes of trapped and non-trapped PARP1. These analyses identified an interaction between trapped PARP1 and the ubiquitin-regulated p97 ATPase/segregase. We found that following trapping, PARP1 is SUMOylated by PIAS4 and subsequently ubiquitylated by the SUMO-targeted E3 ubiquitin ligase RNF4, events that promote recruitment of p97 and removal of trapped PARP1 from chromatin. Small-molecule p97-complex inhibitors, including a metabolite of the clinically used drug disulfiram (CuET), prolonged PARP1 trapping and enhanced PARP inhibitor-induced cytotoxicity in homologous recombination-defective tumour cells and patient-derived tumour organoids. Together, these results suggest that p97 ATPase plays a key role in the processing of trapped PARP1 and the response of tumour cells to PARP inhibitors.

A Hierarchical Error Correction Strategy for Text DNA Storage.

DNA storage has been a thriving interdisciplinary research area because of its high density, low maintenance cost, and long durability for information storage. However, the complexity of errors in DNA sequences including substitutions, insertions and deletions hinders its application for massive data storage. Motivated by the divide-and-conquer algorithm, we propose a hierarchical error correction strategy for text DNA storage. The basic idea is to design robust codes for common characters which have one-base error correction ability including insertion and/or deletion. The errors are gradually corrected by the codes in DNA reads, multiple alignment of character lines, and finally word spelling. On one hand, the proposed encoding method provides a systematic way to design storage friendly codes, such as 50% GC content, no more than 2-base homopolymers, and robustness against secondary structures. On the other hand, the proposed error correction method not only corrects single insertion or deletion, but also deals with multiple insertions or deletions. Simulation results demonstrate that the proposed method can correct more than 98% errors when error rate is less than or equal to 0.05. Thus, it is more powerful and adaptable to the complicated DNA storage applications.

Competitive and Recreational Running Kinematics Examined Using Principal Components Analysis.

Kinematics data are primary biomechanical parameters. A principal component analysis (PCA) of waveforms is a statistical approach used to explore patterns of variability in biomechanical curve datasets. Differences in experienced and recreational runners' kinematic variables are still unclear. The purpose of the present study was to compare any differences in kinematics parameters for competitive runners and recreational runners using principal component analysis in the sagittal plane, frontal plane and transverse plane. Forty male runners were divided into two groups: twenty competitive runners and twenty recreational runners. A Vicon Motion System (Vicon Metrics Ltd., Oxford, UK) captured three-dimensional kinematics data during running at 3.3 m/s. The principal component analysis was used to determine the dominating variation in this model. Then, the principal component scores retained the first three principal components and were analyzed using independent t-tests. The recreational runners were found to have a smaller dorsiflexion angle, initial dorsiflexion contact angle, ankle inversion, knee adduction, range motion in the frontal knee plane and hip frontal plane. The running kinematics data were influenced by running experience. The findings from the study provide a better understanding of the kinematics variables for competitive and recreational runners. Thus, these findings might have implications for reducing running injury and improving running performance.

DNMTs Play an Important Role in Maintaining the Pluripotency of Leukemia Inhibitory Factor-Dependent Embryonic Stem Cells.

Naive pluripotency can be maintained in medium with two inhibitors plus leukemia inhibitory factor (2i/LIF) supplementation, which primarily affects canonical WNT, FGF/ERK, and JAK/STAT3 signaling. However, whether one of these three supplements alone is sufficient to maintain naive self-renewal remains unclear. Here we show that LIF alone in medium is sufficient for adaptation of 2i/L-ESCs to embryonic stem cells (ESCs) in a hypermethylated state (L-ESCs). Global transcriptomic analysis shows that L-ESCs are close to 2i/L-ESCs and in a stable state between naive and primed pluripotency. Notably, our results demonstrate that DNA methyltransferases (DNMTs) play an important role in LIF-dependent mouse ESC adaptation and self-renewal. LIF-dependent ESC adaptation efficiency is significantly increased in serum treatment and reduced in Dnmt3a or Dnmt3l knockout ESCs. Importantly, unlike epiblast stem cells, L-ESCs contribute to somatic tissues and germ cells in chimeras. L-ESCs cultured under such simple conditions as in this study would provide a more conducive platform to clarify the molecular mechanism of ESCs in in vitro culture.

The biomechanical characteristics of a feline distal forelimb: A finite element analysis study.

As a typical digitigrade mammal, the uniquely designed small distal limbs of the feline support two to three times of its body weight during daily movements. To understand how force transmission occurs in relation to the distal joint in a feline limb, which transfers bodyweight to the ground, it is necessary to examine the internal stress distribution of the distal joint limb in detail. Therefore, finite element models (FEM) of a healthy feline were established to predict the internal stress distribution of the distal limb. The FEM model included 23 bony components, various cartilaginous ligaments, as well as the encapsulated soft tissue of the paw. The FEM model was validated by comparison of paw pressure distribution, obtained from an experiment for balance standing. The results demonstrated a good agreement between the experimentally measured and numerically predicted pressure distribution in the feline paw. Additionally, higher stress levels were noted in the metacarpal segment, with smaller stresses observed in the phalanges portion including the proximal, middle, and distal segments. The raised metacarpal segment plays an important role in creating a stiff junction between the metacarpophalangeal (MCP) and wrist joint, stabilizing the distal limb. The paw pads help to optimize stress distribution in phalanx region. Findings from this study contribute to our understanding of feline distal forelimb biomechanical behavior. This information can be applied to bionic design of footwear since an optimal stiff junction and pressure distribution can be adapted to enhance injury relief and sports activities. Further developments may include progress, evaluation, and treatment of metatarsophalangeal joint injuries in human populations.

The Influence of Heel Height on Strain Variation of Plantar Fascia During High Heel Shoes Walking-Combined Musculoskeletal Modeling and Finite Element Analysis.

The therapeutic benefit of high heel shoes (HHS) for plantar fasciitis treatment is controversial. It has been suggested that plantar fascia strain can be decreased by heel elevation of shoes which helps in body weight redistribution throughout the length of the foot. Yet it is a fact that the repetitive tension caused by HHS wearing resulting in plantar fasciitis is a high-risk disease in HHS individuals who suffer heel and plantar pain. To explore the biomechanical function on plantar fascia under HHS conditions, in this study, musculoskeletal modeling (MsM) and finite element method (FEM) were used to investigate the effect of heel height on strain distribution of plantar fascia. Three-dimensional (3D) and one-dimensional (1D) finite element models of plantar fascia were generated to analyze the computed strain variation in 3-, 5-, and 7-cm heel heights. For validation, the computed foot contact pressure was compared with experimental measurement, and the strain value on 1D fascia was compared with previous studies. Results showed that the peak strain of plantar fascia was progressively increased on both 3D and 1D plantar fascia as heel elevated from 3 to 7 cm, and the maximum strain of plantar fascia occurs near the heel pain site at second peak stance. The 3D fascia model predicted a higher strain magnitude than that of 1D and provided a more reliable strain distribution on the plantar fascia. It is concluded that HHS with narrow heel support could pose a high risk on plantar fasciitis development, rather than reducing symptoms. Therefore, the heel elevation as a treatment recommendation for plantar fasciitis is questionable. Further studies of different heel support structures of shoes to quantify the effectiveness of heel elevation on the load-bearing mechanism of plantar fascia are recommended.

A Mixed Comparisons of Aerobic Training With Different Volumes and Intensities of Physical Exercise in Patients With Hypertension: A Systematic Review and Network Meta-Analysis.

It is essential for patients with hypertension to effectively reduce and maintain appropriate blood pressure levels. As one of the non-pharmacological and invasive methods, physical exercise seems to improve blood pressure of the patients with hypertension. However, different volumes and intensities of physical exercise on the improvement of hypertension are different. To understand the effects of the type of exercise training on blood pressure and the other health status of patients with hypertension, a network meta-analysis was used to compare the mixed effects of different types of exercise training. This systematic review includes all eligible randomized controlled trials of PubMed, Medline, Cochrane Library, and CINAHL. Twelve studies met the inclusion criteria (n = 846 participants at the end of the study). The results show that a medium-intensity training (MIT) is best in improving the blood pressure of patients with hypertension, while a high-volume high-intensity interval training (HVHIIT) is better in reducing body mass and resting heart rate. In addition, the analysis of the exercise capacity shows that HVHIIT has a better effect on the improvement of patients with hypertension. Noticeably, long-term high-volume and appropriate intensity exercise can effectively improve the health status of patients with hypertension. In short, for patients with high blood pressure, MIT seems to be better at lowering blood pressure, while HVHIIT can better improve exercise ability and physical fitness. However, larger randomized controlled trials with a longer duration than those included in this meta-analysis are needed to confirm these results.

Solid inoculants as a practice for bioaugmentation to enhance bioremediation of hydrocarbon contaminated areas.

Vacuum freeze-drying is a scientifically advanced method to prepare solid inoculants from oil degrading bacterium. The introduction of oil-degrading microbes or bioaugmentation can be an efficient way to bioremediate oil spills in marine areas, where oil-degrading bacteria are deficient. The purpose of this study is to evaluate the potential use of solid inoculants of LZ-2 bacteria to enhance the degradation rate of crude oil. In this study, response surface methodology (RSM) was incorporated into the experimental design to optimize a response, which is influenced by different protectants. Our results showed that five factors have interactive and synergistic protective effects on the growth of LZ-2. Optimal growth of freeze-dried LZ-2 (63.8%) was observed with a 10.5% solution of skim milk supplemented with 14.3% sucrose, 14.4% of trehalose, 4.9% of glycerin and 14.7% of ß-cyclodextrin. The culture grew in medium containing crude oil (3 g L-1) at 37 °C at 150 rpm for 30 d, GC and GC-MS analysis showed biodegradation of 44.2 and 21.6% for total saturate and aromatic hydrocarbons respectively. These results indicated that the solid inoculants of LZ-2 bacteria had the potential to be used for ex-situ bioremediation of hydrocarbon pollutants associated with crude oil.

TEX264 coordinates p97- and SPRTN-mediated resolution of topoisomerase 1-DNA adducts.

Eukaryotic topoisomerase 1 (TOP1) regulates DNA topology to ensure efficient DNA replication and transcription. TOP1 is also a major driver of endogenous genome instability, particularly when its catalytic intermediate-a covalent TOP1-DNA adduct known as a TOP1 cleavage complex (TOP1cc)-is stabilised. TOP1ccs are highly cytotoxic and a failure to resolve them underlies the pathology of neurological disorders but is also exploited in cancer therapy where TOP1ccs are the target of widely used frontline anti-cancer drugs. A critical enzyme for TOP1cc resolution is the tyrosyl-DNA phosphodiesterase (TDP1), which hydrolyses the bond that links a tyrosine in the active site of TOP1 to a 3' phosphate group on a single-stranded (ss)DNA break. However, TDP1 can only process small peptide fragments from ssDNA ends, raising the question of how the ~90 kDa TOP1 protein is processed upstream of TDP1. Here we find that TEX264 fulfils this role by forming a complex with the p97 ATPase and the SPRTN metalloprotease. We show that TEX264 recognises both unmodified and SUMO1-modifed TOP1 and initiates TOP1cc repair by recruiting p97 and SPRTN. TEX264 localises to the nuclear periphery, associates with DNA replication forks, and counteracts TOP1ccs during DNA replication. Altogether, our study elucidates the existence of a specialised repair complex required for upstream proteolysis of TOP1ccs and their subsequent resolution.

Activin A and BMP4 Signaling Expands Potency of Mouse Embryonic Stem Cells in Serum-Free Media.

Inhibitors of Mek1/2 and Gsk3ß, known as 2i, and, together with leukemia inhibitory factor, enhance the derivation of embryonic stem cells (ESCs) and promote ground-state pluripotency (2i/L-ESCs). However, recent reports show that prolonged Mek1/2 suppression impairs developmental potential of ESCs, and is rescued by serum (S/L-ESCs). Here, we show that culturing ESCs in Activin A and BMP4, and in the absence of MEK1/2 inhibitor (ABC/L medium), establishes advanced stem cells derived from ESCs (esASCs). We demonstrate that esASCs contributed to germline lineages, full-term chimeras and generated esASC-derived mice by tetraploid complementation. We show that, in contrast to 2i/L-ESCs, esASCs display distinct molecular signatures and a stable hypermethylated epigenome, which is reversible and similar to serum-cultured ESCs. Importantly, we also derived novel ASCs (blASCs) from blastocysts in ABC/L medium. Our results provide insights into the derivation of novel ESCs with DNA hypermethylation from blastocysts in chemically defined medium.

The p97-Ataxin 3 complex regulates homeostasis of the DNA damage response E3 ubiquitin ligase RNF8.

The E3 ubiquitin ligase RNF8 (RING finger protein 8) is a pivotal enzyme for DNA repair. However, RNF8 hyper-accumulation is tumour-promoting and positively correlates with genome instability, cancer cell invasion, metastasis and poor patient prognosis. Very little is known about the mechanisms regulating RNF8 homeostasis to preserve genome stability. Here, we identify the cellular machinery, composed of the p97/VCP ubiquitin-dependent unfoldase/segregase and the Ataxin 3 (ATX3) deubiquitinase, which together form a physical and functional complex with RNF8 to regulate its proteasome-dependent homeostasis under physiological conditions. Under genotoxic stress, when RNF8 is rapidly recruited to sites of DNA lesions, the p97-ATX3 machinery stimulates the extraction of RNF8 from chromatin to balance DNA repair pathway choice and promote cell survival after ionising radiation (IR). Inactivation of the p97-ATX3 complex affects the non-homologous end joining DNA repair pathway and hypersensitises human cancer cells to IR. We propose that the p97-ATX3 complex is the essential machinery for regulation of RNF8 homeostasis under both physiological and genotoxic conditions and that targeting ATX3 may be a promising strategy to radio-sensitise BRCA-deficient cancers.

[Immunogenicity of the truncated NDV F protein surface-displayed on Lactobacillus casei].

To evaluate immune efficacy of the recombinant Lactobacillus casei, we constructed pLA-Newcastle disease virus (NDV)-F/L. casei and obtained the expression products. PCR amplified the NDV F gene carrying part of the major epitopes. The target gene was inserted to the shuttle plasmid pLA, and then transformed into Escherichia coli BL21 (DE3) in order to screen positive recombinant plasmid. The positive recombinant plasmid was transformed into L. casei by electroporation to construct pLA-NDV-F/L. casei. The positive strains were identified by PCR. The reactivity of the recombinant bacteria was identified by Western blotting and the protein expression was detected by indirect immunofluorescence, flow cytometry and laser confocal microscopy. The 14-day-old chickens in each group were vaccinated by oral plus nose drops. The pLA-NDV-F/L. casei twice immunization group and three times immunization group, the commercial vaccine group, the pLA/L. casei group, the unchallenge PBS and the challenge PBS group were established. IgG in serum and sIgA in the lavage fluid of intestinal, nasal and lung were detected by ELISA. The protection rate of chickens was evaluated. The results showed that 94.10% of the recombinant bacteria expressed the F protein. The recombinant protein was highly expressed on the surface of L. casei with a protein size of 62 kDa, which specifically bound to anti-NDV serum. The levels of anti-F IgG and sIgA antibodies in each test group were significantly higher than those in the control groups. The duration of antibody in the pLA-NDV-F/L. casei three-time immunization group lasted 28 days longer than that in the twice immunized group, and there was no significant difference between antibody peak values. The attack protection rates in each group of immunized pLA-NDV-F/L. casei three times, twice, attenuated vaccine, pLA/L. casei and PBS were 80%, 80%, 90%, 0% and 0%, respectively. Therefore, the antigenic protein of NDV F was successfully expressed by L. casei expression system, which has of reactogenicity and immunogenicity, and could induce protective immune responses in chickens.