An efficient segmentation model for abnormal chicken droppings recognition based on improved deep dual-resolution network.

The characteristics of chicken droppings are closely linked to their health status. In prior studies, chicken droppings recognition is treated as an object detection task, leading to challenges in labeling and missed detection due to the diverse shapes, overlapping boundaries, and dense distribution of chicken droppings. Additionally, the use of intelligent monitoring equipment equipped with edge devices in farms can significantly reduce manual labor. However, the limited computational power of edge devices presents challenges in deploying real-time segmentation algorithms for field applications. Therefore, this study redefines the task as a segmentation task, with the main objective being the development of a lightweight segmentation model for the automated monitoring of abnormal chicken droppings. A total of 60 Arbor Acres broilers were housed in 5 specific pathogen-free cages for over 3 wk, and 1650 RGB images of chicken droppings were randomly divided into training and testing sets in an 8:2 ratio to develop and test the model. Firstly, by incorporating the attention mechanism, multi-loss function, and auxiliary segmentation head, the segmentation accuracy of the DDRNet was enhanced. Then, by employing the group convolution and an advanced knowledge-distillation algorithm, a lightweight segmentation model named DDRNet-s-KD was obtained, which achieved a mean Dice coefficient (mDice) of 79.43% and an inference speed of 86.10 frames per second (FPS), showing a 2.91% and 61.2% increase in mDice and FPS compared to the benchmark model. Furthermore, the DDRNet-s-KD model was quantized from 32-bit floating-point values to 8-bit integers and then converted to TensorRT format. Impressively, the weight size of the quantized model was only 13.7 MB, representing an 82.96% reduction compared to the benchmark model. This makes it well-suited for deployment on the edge device, achieving an inference speed of 137.51 FPS on Jetson Xavier NX. In conclusion, the methods proposed in this study show significant potential in monitoring abnormal chicken droppings and can provide an effective reference for the implementation of other agricultural embedded systems.

The characteristics of chicken droppings are closely related to their health. In this study, we developed a lightweight segmentation model for chicken droppings and evaluated its inference speed on the edge device with limited computational power. The results showed that the proposed model exhibits significant potential in the early warning of abnormal chicken droppings, which can help producers implement interventions before disease outbreaks, thereby avoiding great economic losses. Additionally, the model optimization and compression processes proposed in this study can provide an effective reference for the implementation of other embedded systems.

Utilizing Machine Learning to Identify Biomarkers of Endoplasmic Reticulum Stress and Analyze Immune Cell Infiltration in Parkinson's Disease.

The neurodegenerative disorder known as Parkinson's disease (PD) affects many people. The objective of this investigation was to examine the relationship between immune system infiltration, ATP-binding cassette transporter subfamily A member 7 (ABCA7) and TBL2 as well as potential therapeutic targets for the identification of PD associated to endoplasmic reticulum (ER) stress. First, we obtained PD data through GEO and divided it into two sets: a training set (GSE8397) plus a set for validation (GSE7621). Functional enrichment analysis was performed on a set of DEGs that overlapped with genes involved in endoplasmic reticulum stress. To identify genes of PD linked with endoplasmic reticulum stress, we employed random forest (RF) along with the least absolute shrinkage and selection operator (LASSO) logistic regression. Spearman's rank correlation analysis was then used to find associations among diagnostic markers with immune cell penetration. A grand total of 2 stress-related endoplasmic reticulum signature transcripts were identified. ABCA7 and TBL2 were shown to have diagnostic potential for PD and immune infiltrating cells have a role in the etiology of the disease. Additionally, resting CD4 memory, plasma cells, and NK cells overall exhibited positive associations with ABCA7, whereas triggered macrophages, T cells with active CD4 memory, activating NK cells, T cells with activated CD4 naive, engaged NK cells, and neutrophils all had adverse interactions with ABCA7. Overall, ABCA7 together with TBL2 have diagnostic utility for PD, and several types of immune cells, especially macrophages, may be involved in the development and progression of the disease.

Enhancing Oral Mucosal Barrier, Mitigating Microinflammation, and Addressing Malnutrition in Dialysis Patients: The Impact of Tangerine Peel Lemon Glycerin.

Objective: This study investigates the efficacy of tangerine peel lemon glycerin extract oral spray in improving oral mucosal barrier, reducing microinflammation, and addressing malnutrition in maintenance dialysis (MHD) patients. Methods: Tangerine peel and dry lemon underwent glycerin extraction. From January 2021 to June 2022, 72 MHD patients with thirst were prospectively chosen at Sinopharm Gezhouba Central Hospital. Randomization divided them into an observation group (n=36) and a control group (n=36). Both received routine maintenance dialysis and chronic kidney disease management. Oral conditions were assessed using OHIP-14, a homemade visual thirst score scale, SFR, sAA, and saliva pH. Microinflammatory indexes (CRP, TNF-α, IL-6) and nutritional status indicators (Alb, PA, Hb) were measured. The observation group used 1ml of tangerine peel lemon glycerin extract with a pH value of 5.9~6.1 q6h, while the control group used 1ml of purified water q6h. Results: After 3 months, the observation group showed significant improvement in OHIP-14 and visual thirst score scale (P < .01). Saliva pH, CRP, TNF-α, and IL-6 levels decreased, and SAA activity, SFR, Alb, PA, and Hb levels increased significantly in the observation group compared to the control group (P < .01). Conclusions: Tangerine peel lemon glycerin spray demonstrates promise in improving the oral mucosal barrier, exhibiting antibacterial and anti-inflammatory properties that mitigate microinflammation and malnutrition. This finding suggests a connection between oral health, systemic pathology, psychological state, and social adaptability.

ß-cyclocitral, a novel AChE inhibitor, contributes to the defense of Microcystis aeruginosa against Daphnia grazing.

ß-cyclocitral is one of the major compounds in cyanobacterial volatile organic compound (VOCs) and can poison other aquatic organisms. To investigate the effect of ß-cyclocitral on cyanobacterial-grazer interactions, Daphnia sinensis was fed Microcystis aeruginosa and exposed to ß-cyclocitral. Our present study demonstrated that M. aeruginosa could significantly inhibit D. sinensis grazing. And the grazing inhibition by Microcystis aeruginosa results from the suppression of feeding rate, heart rate, thoracic limb activity and swimming speed of D. sinensis. In addition, M. aeruginosa could also induce intestinal peristalsis and emptying in D. sinensis. Interestingly, our present study found that the exposure to ß-cyclocitral could mimic a range of phenotypes induced by M. aeruginosa in D. sinensis. These results suggested that M. aeruginosa could release ß-cyclocitral to inhibit Daphnia grazing. To further examine the toxic mechanism of ß-cyclocitral in Daphnia, several in vivo and in vitro experiments displayed that ß-cyclocitral was a novel inhibitor of acetylcholinesterase (AChE). It could induce the accumulation of acetylcholine (ACh) by inhibiting AchE activity in D. sinensis. High level of endogenous Ach could inhibit feeding rate and induce intestinal peristalsis and emptying in D. sinensis.

In Situ Preparation of Three-Dimensional Porous Nickel Sulfide as a Battery-Type Supercapacitor.

A one-step sulfurization method to fabricate Ni3S2 nanowires (Ni3S2 NWs) directly on a Ni foam (NF) was developed as a simple, low-cost synthesis method for use as a supercapacitor (SC), aimed at optimizing energy storage. Ni3S2 NWs have high specific capacity and are considered a promising electrode material for SCs; however, their poor electrical conductivity and low chemical stability limit their applications. In this study, highly hierarchical three-dimensional porous Ni3S2 NWs were grown directly on NF by a hydrothermal method. The feasibility of the use of Ni3S2/NF as a binder-free electrode for achieving high-performance SCs was examined. Ni3S2/NF exhibited a high specific capacity (255.3 mAh g-1 at a current density of 3 A g-1), good rate capability (2.9 times higher than that of the NiO/NF electrode), and competitive cycling performance (capacity retention of specific capacity of 72.17% after 5000 cycles at current density of 20 A g-1). Owing to its simple synthesis process and excellent performance as an electrode material for SCs, the developed multipurpose Ni3S2 NWs electrode is expected to be a promising electrode for SC applications. Furthermore, the synthesis method of self-growing Ni3S2 NW electrodes on 3D NF via hydrothermal reactions could potentially be applied to the fabrication of SC electrodes using a variety of other transition metal compounds.

Eliciting effective tumor immunity against ovarian cancer by cancer stem cell vaccination.

Advanced ovarian cancer (OC) patients have limited benefit from current relevant cytotoxic and targeted therapies following debulking surgery. Therefore, new therapeutic strategies are in urgent need. Immunotherapy has shown great potential in tumor treatment, especially in tumor vaccine development. The study objective was to evaluate the immune effects of cancer stem cells (CSCs) vaccines on OC. The CD44+CD117+CSCs were isolated from human OC HO8910 and SKOV3 cells using the magnetic cell sorting system; the cancer stem-like cells were selected from murine OC ID8 cell by no-serum formed sphere culture. The CSC vaccines were prepared by freezing and thawing these CSCs, which were then injected into mice followed by challenging the different OC cells. The in vivo antitumor efficacy of CSC immunization revealed the vaccines were capable of significantly provoking immune responses to autologous tumor antigens in vaccinated mice as the mice were found to have markedly inhibited tumor growth, prolonged survival, and decreased CSC counts in OC tissues when compared to mice without the CSC vaccination. The in vitro cytotoxicities of immunocytes toward SKOV3, HO8910 and ID8 cells indicated a significant killing efficacy compared with the controls. However, the antitumor efficacy was remarkably reduced whilst the mucin-1 expression in CSC vaccines was down-regulated by small interfering RNA. Overall, findings from this study provided the evidence that has deepened our understanding of CSC vaccine immunogenicity and anti-OC efficacy, particularly for the role of dominant antigen mucin-1. It is possible to turn the CSC vaccine into an immunotherapeutic approach against ovarian cancer.

Clinical characteristics of catheter-related infection in patients with chronic renal failure End Stage Renal failure undergoing semi-permanent catheter placement during maintenance hemodialysis through tunnelled cuffed hemodialysis catheter.

Objective: To analysis the relevant infections and risk factors of patients undergoing hemodialysis semi-permanent catheter (tunneled cuffed) placement during for maintenance hemodialysis. Methods: A total of 158 patients with chronic renal failure (CRF) End stage renal failure (ESRF) treated in our hospital from September 2018 to September 2021 were retrospectively analyzed. All the patients underwent semi-permanent catheter placement during maintenance hemodialysis. The occurrence of catheter-related infections in the patients were recorded. The patients with catheter-related infections were included in the infection group, and the others without infection in the non-infection group. The differences in hypertension, gender, diabetes, age, catheter indwelling time and dialysis time between the two groups were analyzed, and the distribution of pathogens in the patients with infections was analyzed. Results: The patients were followed up for 13 to 36 months, with an average of (22.18 ± 6.09) months. Among the 158 patients who underwent going semi-permanent catheter placement, 42 (26.58%) presented semi-permanent catheter-related infections, including four cases of catheter-related bacteremia, 16 cases of tunnel infection and 22 cases of catheter exit-site infection. Among total of 42 strains of pathogens were isolated from the 42 patients with catheter-related infections, including 243 strains of Gram-positive cocci were identified in 24/42(57.14%), and 163 strains of Gram-negative bacilli were identified 16/42(38.10%) and one starin of fungus was identified in 2/42 patients. Statistically significant differences were found in dialysis duration time, hypoalbuminemia, average mean age, diabetes and catheter indwelling time between patients with and without catheter-related infections (P < 0.05). Hypoalbuminemia, catheter indwelling time and diabetes were risk factors for catheter-related infections (P < 0.05). Conclusions: Patients with ESRF CRF are at risk and prone to catheter-related infections during hemodialysis using catheter, mainly tunnel infection and catheter exit-site infection. Gram-positive cocci are the main pathogens. Hypoalbuminemia, too long catheter indwelling time and diabetes are the risk factors for infections.

Study on Continuous Hydrolysis in a Microchannel Reactor for the Production of Titanium Dioxide by a Sulfuric Acid Process.

The development of a continuous hydrolysis process of titanium sulfate is an innovation to the traditional production process of titanium dioxide by the sulfuric acid process. In the experiment, a microchannel reactor was designed, and the hydrolysis rate of titanium sulfate, the particle size, and particle size distribution of metatitanic acid agglomerates were used as indicators to investigate the effect of operating conditions on the continuous hydrolysis of titanium sulfate. The results have shown that as the amount of dilution water increased, the hydrolysis rate of titanium sulfate decreased, and the particle size of primary aggregates of metatitanic acid increased from 39 to 54 nm. As the alkali mass concentration of dilution water increased, the hydrolysis rate of titanyl sulfate increased, and the particle size of primary aggregates of metastatic acid first decreased and then increased, and the particle size range was 40-48 nm. As the flow rate increased, the hydrolysis rate of titanyl sulfate increased, and the particle size of primary aggregates of metatitanic acid dropped from 59 to 43 nm. Compared with the batch hydrolysis operation, the continuous process has stronger anti-disturbance ability, significantly shorter operation time of the reaction section, and narrower particle size distribution of the product metatitanic acid.

Chemical Profiling of Kaliziri Injection and Quantification of Six Caffeoyl Quinic Acids in Beagle Plasma by LC-MS/MS.

Vitiligo is a stubborn multifactorial skin disease with a prevalence of approximately 1% in the global population. Kaliziri, the seeds of Vernonia anthelmintica (L.) Willd., is a well-known traditional Uyghur medicine for the treatment of vitiligo. Kaliziri injections is a Chinese-marketed treatment approved by the China Food and Drug Administration for the treatment of vitiligo. The significant effects of Kaliziri injection have been thoroughly studied. However, chemical components studies and plasma quantification studies are lacking for Kaliziri injection. Ultra-high-performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry was employed to comprehensively characterize the caffeoyl quinic acid derivatives present in Kaliziri injection. Based on accurate mass measurements, key fragmental ions and comparisons with reference standards, 60 caffeoyl quinic acid derivatives were identified in Kaliziri injections, including caffeoyl quinic acids, coumaroyl caffeoyl quinic acids, dicaffeoyl quinic acids, feruloyl caffeoyl quinic acids, and dicaffeoyl quinic acid hexosides. Moreover, an HPLC-MS/MS method was developed and validated for the quantitative analysis of 5-caffeoyl quinic acid, 4-caffeoyl quinic acid, 1,3-dicaffeoyl quinic acid, 3,4-dicaffeoyl quinic acid, 3,5-dicaffeoyl quinic acid and 4,5-dicaffeoyl quinic acid in beagle plasma. The quantitative HPLC-MS/MS method was applied to quantify these six major caffeoyl quinic acids in beagle plasma after the subcutaneous administration of Kaliziri injection. All of the six analytes reached their peak plasma of concentrations within 30 min.

Elevated IL-35 level and iTr35 subset increase the bacterial burden and lung lesions in Mycobacterium tuberculosis-infected mice.

This study aimed to investigate the relationship between interleukin (IL)-35 level and IL-35-producing regulatory T cells (iTr35 subset) in Mycobacterium tuberculosis (Mtb)-infected mice. After the mice were injected with Mtb strain H37R via tail vein, the bacterial burden, lung lesions, and the impact of immune suppression on the infected mice were respectively assessed. The results, when compared with the control mice, showed that the mRNA expression levels of the p35 and Epstein-Barr virus-induced gene 3 of IL-35 were significantly increased in the Mtb-infected mouse spleen at 4 or 8 weeks post-infection and their protein expression levels were concurrently increased in the lungs of the mice, especially in 8 week infected mice. In addition, the levels of serum IL-35 and the iTr35 subset in the spleen of mice were also increased in 4 or 8 weeks post-infection compared with the control mice. Importantly, the high bacterial burden and lung lesions and the low mouse weight were found at 8 week post-infection. Therefore, the mice infected with Mtb resulted in elevating IL-35 level and iTr35 subset and increasing bacterial burden and lung lesions. The findings from the study suggest IL-35 and iTr35 cells may exert an immune suppression role in chronic Mtb-infected mice.

Multi-omics perspective on studying reproductive biology in Daphnia sinensis.

Daphnia sinensis is a widespread freshwater microcrustacean. The assembled D. sinensis genome totaled 131.58 Mb with 92.23% of the assembly anchored onto 10 chromosomes. Based on the whole genome information, we further compared the transcriptomic and epigenomic characterization among parthenogenetic females, sexual females and males in D. sinensis. Transcriptomic analysis showed that the up-regulated genes in males were mainly grouped into the cuticle, sex differentiation and methyl farnesoate synthesis, which might play a pivotal role in steering development and reproduction processes. By comparison, the highly expressed genes in parthenogenetic females were mainly grouped into energy metabolism, mitosis, and DNA replication, which might contribute to maintaining rapid production of parthenogenetic females, and nutrient uptake for the growth of neonates. The whole-genome DNA methylation analysis showed that the methylation rate in parthenogenetic females was higher than that in sexual females and males, which might contribute to its rapid response to environment stress.

Mucin 1 downregulation decreases the anti-tumor effects of melanoma vaccine.

Background: Immunotherapy-based approaches are important breakthroughs with potential treatment benefits for melanoma patients. Mucin 1 (MUC1) is significantly upregulated in melanoma relative to normal cells. It has been reported that MUC1 influences cancer cell proliferation, apoptosis, invasion, and metastasis.The study aimed to explore the effect of MUC1 knockdown on the biological characteristics of the melanoma cell line B16F10 and evaluate whether MUC1 is an effective candidate target antigen for melanoma vaccine development. Methods: First, lentiviral vector-mediated short hairpin RNA (shRNA) was used to knockdown MUC1 in B16F10 cells (shMUC1-B16F10 cells). Next, we examined epithelial-mesenchymal transition (EMT), migration, proliferative capacity, clone formation, and distribution of cell cycle in shMUC1-B16F10 cells. Finally, the vaccine was prepared by repeated freeze-thawing of the shMUC1-B16F10 cells and used to subcutaneously immunize C57BL/6 mice, which were then challenged using B16F10 cells 10 days after the final vaccination. Results: It was revealed that shMUC1 suppressed B16F10 proliferative and colony formation capacity, induced the arrest of cell cycle in the G0/G1 phase, and adjusted the expression of EMT-associated factors. MUC1 downregulation markedly suppressed the effect of B16F10 vaccine against melanoma in a mouse model. As compared with B16F10-vaccinated mice, B16F10-vaccinated mice in which MUC1 was silenced had reduced natural killer (NK) cytotoxicity, lower production of interferon-γ (IFN-γ), anti-MUC1 antibodies, perforin, granzyme B, and elevated tumor growth factor-ß (TGF-ß) level. Conclusions: MUC1 has strong melanoma vaccine immunogenicity, and induces the host's anti-tumor reaction. MUC1 knockdown inhibits the immune activity of B16F10 cell vaccine and anti-melanoma effect, suggesting the MUC1 is an important candidate target antigen of the melanoma vaccine.

Knockdown of ALDH1A3 reduces breast cancer stem cell marker CD44 via the miR-7-TGFBR2-Smad3-CD44 regulatory axis.

Inhibition of aldehyde dehydrogenase 1 family member A3 (ALDH1A3) has been revealed to lead to significant increase of microRNA (miR)-7 expression and decrease of CD44 expression in breast cancer stem cells (BCSCs), however the mechanism is not clear. The aim of the present study was to investigate the regulatory relationship between ALDH1A3, miR-7, and CD44 in BCSCs. The expression of ALDH1A3 was inhibited by small interfering RNA (siRNA or si), and the expression of miR-7 was detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Then, the ratio of CD44+ cells was analyzed by flow cytometry in MDA-MB-231 cells. The dual-luciferase reporter system was used to demonstrate that miR-7 binds to transforming growth factor-ß receptor 2 (TGFBR2) 3'UTR, and ChIP-PCR determined whether the transcription factor Smad3 binds to the upstream regulatory region of the CD44 promoter. The results revealed that siALDH1A3 downregulated ALDH1A3 and promoted miR-7 expression, which resulted in downregulation of CD44 expression. siALDH1A3 also downregulated the CD44 expression on the surface of MDA-MB-231 cells and inhibited the G2/M phase in BCSCs as analyzed by flow cytometry. In addition, lenti-miR-7 cells transfected with TGF-ß1 + SB431542 revealed that lenti-miR-7 inhibited the TGF-ß1 pathway by inhibiting Smad2/3/4 expression and, thus, downregulated CD44 expression. miR-7 was revealed to directly bind to the TGFBR2 3'UTR through dual-luciferase reporter assay, and Smad3, a transcription factor, through ChIP-PCR was demonstrated to bind to the upstream region of the CD44 promoter. These results demonstrated the existence of the ALDH1A3-miR-7-TGFBR2-Smad3-CD44 axis in MDA-MB-231 cells. RT-qPCR results of 12 breast cancer surgical specimens and SK-BR-3, MCF-7, and LD cell lines further confirmed the presence of the regulatory axis. In conclusion the findings from the present study demonstrated that the ALDH1A3-miR-7-TGFBR2-Smad3-CD44 regulatory axis was highly efficient in the inhibition of CD44 expression in BCSCs, and that the regulatory expression of ALDH1A3 and miR-7 may provide a strategy in the therapy of breast cancer.

Effects of different chemical modifications on the structure and biological activities of polysaccharides from Orchis chusua D. Don.

In this study, an enzyme-assisted extraction method was used to extract Orchis chusua D. Don (Salep) polysaccharide (SP), which was then modified by sulfation, acetylation, phosphorylation, and carboxymethylation to obtain modified polysaccharides. Furthermore, their degree of substitution, chemical composition, and molecular weight were evaluated. The primary structural features were characterized by UV spectra, FT-IR spectra, Congo-red test, and scanning electron microscope. The phosphorylated polysaccharide (SP-P) was demonstrated the highest scavenging ability on hydroxyl radical and growth-promoting activity on Lactobacillus Bulgaricus. The carboxymethylated polysaccharide (SP-C) was exhibited the strongest DPPH and ABTS radical scavenging effects. The acetylated polysaccharide (SP-A) displayed the best proliferation effects on Bifidobacterium adolescentis, whereas the sulfated polysaccharide (SP-S) maintained moderately stable antioxidant and probiotic ability. These findings indicate that the modified polysaccharides had their potential significance as new antioxidants and probiotics for the food industry. PRACTICAL APPLICATION: This article provides a new source for the development of polysaccharide derivatives as new antioxidants and probiotics for the food industry.

Engineered endolysin-based "artilysins" for controlling the gram-negative pathogen Helicobacter pylori.

Helicobacter pylori infection can cause a variety of gastrointestinal diseases. In severe cases, there is a risk of gastric cancer. Antibiotics are often used for clinical treatment of H. pylori infections. However, because of antibiotic overuse in recent years and the emergence of multidrug-resistant bacteria, there is an urgent need to develop new treatment methods and drugs to achieve complete eradication of H. pylori. Endolysins and holins encoded by bacterial viruses (i.e., phages) represent a promising avenue of investigation. These lyase-based antibacterial drugs act on the bacterial cell wall to destroy the bacteria. Currently, a type of endolysin that has been studied more frequently acts on the amide bond between peptidoglycans, and holin is a transmembrane protein that can punch holes in the cell membrane. However, as a Gram-negative bacterium, H. pylori possesses a layer of impermeable lipopolysaccharides on the cell wall, which prevents endolysin interaction with the cell wall. Therefore, we designed a genetic linkage between an endolysin enzyme and a holin enzyme with a section of polypeptides (e.g., polycations and hydrophobic peptides) that enable penetration of the outer membrane. These complexes were designated "artilysins" and were efficiently expressed in Escherichia coli. In vitro bacteriostasis experiments showed that the purified artilysins had strong bacteriostatic effects on H. pylori. In addition, the surface of H. pylori was perforated and destroyed, as confirmed by electron microscopy, which was proved that artilysins had bacteriolytic effect on H. pylori.

Iron Oxide Nanoparticles Combined with Cytosine Arabinoside Show Anti-Leukemia Stem Cell Effects on Acute Myeloid Leukemia by Regulating Reactive Oxygen Species.

BACKGROUND AND AIM: Acute myeloid leukemia (AML), initiated and maintained by leukemia stem cells (LSCs), is often relapsed or refractory to therapy. The present study aimed at assessing the effects of nanozyme-like Fe3O4 nanoparticles (IONPs) combined with cytosine arabinoside (Ara-C) on LSCs in vitro and in vivo. METHODS: The CD34+CD38-LSCs, isolated from human AML cell line KG1a by a magnetic activated cell sorting method, were treated with Ara-C, IONPs, and Ara-C+ IONPs respectively in vitro. The cellular proliferation, apoptosis, reactive oxygen species (ROS), and the related molecular expression levels in LSCs were analyzed using flow cytometry, RT-qPCR, and Western blot. The nonobese diabetic/severe combined immune deficiency mice were transplanted with LSCs or non-LSCs via tail vein, and then the mice were treated with Ara-C, IONPs and IONPs plus Ara-C, respectively. The therapeutic effects on the AML bearing mice were further evaluated. RESULTS: LSCs indicated stronger cellular proliferation, more clone formation, and more robust resistance to Ara-C than non-LSCs. Compared with LSCs treated with Ara-C alone, LSCs treated with IONPs plus Ara-C showed a significant increase in apoptosis and ROS levels that might be regulated by nanozyme-like IONPs via improving the expression of pro-oxidation molecule gp91-phox but decreasing the expression of antioxidation molecule superoxide dismutase 1. The in vivo results suggested that, compared with the AML bearing mice treated with Ara-C alone, the mice treated with IONPs plus Ara-C markedly reduced the abnormal leukocyte numbers in peripheral blood and bone marrow and significantly extended the survival of AML bearing mice. CONCLUSION: IONPs combined with Ara-C showed the effectiveness on reducing AML burden in the mice engrafted with LSCs and extending mouse survival by increasing LSC's ROS level to induce LSC apoptosis. Our findings suggest that targeting LSCs could control the AML relapse by using IONPs plus Ara-C.

LRRC31 inhibits DNA repair and sensitizes breast cancer brain metastasis to radiation therapy.

Breast cancer brain metastasis (BCBM) is a devastating disease. Radiation therapy remains the mainstay for treatment of this disease. Unfortunately, its efficacy is limited by the dose that can be safely applied. One promising approach to overcoming this limitation is to sensitize BCBMs to radiation by inhibiting their ability to repair DNA damage. Here, we report a DNA repair suppressor, leucine-rich repeat-containing protein 31 (LRRC31), that was identified through a genome-wide CRISPR screen. We found that overexpression of LRRC31 suppresses DNA repair and sensitizes BCBMs to radiation. Mechanistically, LRRC31 interacts with Ku70/Ku80 and the ataxia telangiectasia mutated and RAD3-related (ATR) at the protein level, resulting in inhibition of DNA-dependent protein kinase, catalytic subunit (DNA-PKcs) recruitment and activation, and disruption of the MutS homologue 2 (MSH2)-ATR module. We demonstrate that targeted delivery of the LRRC31 gene via nanoparticles improves the survival of tumour-bearing mice after irradiation. Collectively, our study suggests LRRC31 as a major DNA repair suppressor that can be targeted for cancer radiosensitizing therapy.

The surface dominant antigen MUC1 is required for colorectal cancer stem cell vaccine to exert anti-tumor efficacy.

Colorectal cancer (CRC), initiated and maintained by colorectal cancer stem cells (CCSCs), ranks the third most common cancers and has drawn wide attentions worldwide. Therefore, targeting clearance of CCSCs has become an important strategy of CRC immunotherapy. Mucin1 (MUC1) is a tumor-associated cell surface antigen of CRC, but its role in CCSC vaccine remains unclear. In the study, we demonstrated that MUC1 may be a dominant antigen to exert antitumor immunity in CCSC vaccine. First, CCSCs were enriched from CT26 cell line via a serum-free sphere formation approach, and were identified by detecting expression of CD133, ALDH, and ALCAM. Then, the isolated CCSCs were frozen for 30 min and thawed for 30 min to prepare the cell lysate. The specific anti-MUC1 antibody was added to the cell lysate to neutralize the dominant antigen MUC1. Finally, mice were subcutaneously immunized with the cell lysate, followed by a challenge with CT26 cells at one week after final vaccination. Attractively, CCSC vaccine significantly activated the NK cells, T cells, and B cells, resulting in inhibiting the tumor growth via a target killing of CCSCs as evidenced by a decrease of CD133+cells in tumor compared to CCSC vaccine with specific anti-MUC1 antibody. In addition, CCSC vaccine reduced expression of inflammatory factors in vaccinated mice. As expected, neutralizing antibody against MUC1 significantly impaired the antitumor efficacy of CCSC vaccine. Overall, CCSC vaccine could serve as a potent vaccine for CRC immunotherapy. The surface dominant antigen MUC1 may play a key role in regulating immunogenicity of CCSCs.

Colorectal cancer stem cell vaccine with high expression of MUC1 serves as a novel prophylactic vaccine for colorectal cancer.

Targeted clearance of colorectal cancer stem cells (CCSCs) has become a novel strategy for tumor immunotherapy. Molecule mucin1 (MUC1) is one of targetable cell surface antigens in CCSCs. However, the critical role of MUC1 in anti-tumor effects of CCSC vaccine remains unclear. In the present study, we showed that MUC1 may be required for CCSC vaccine to exert tumor immunity. CD133+CCSCs were isolated from CT26 cell line using a magnetic-activated cell sorting system, and MUC1 shRNA or recombinant plasmid was further used to decrease or increase the expression of MUC1 in CD133+CCSCs. Mice were subcutaneously immunized with the CCSC lysates, MUC1 knockin CCSCs, and MUC1 knockdown CCSCs respectively, followed by a challenge with CT26 cells. We found that CCSC vaccine significantly reduced the tumor growth via a target killing of CCSCs as evidenced by a decrease of CD133+ cells and ALDH+ cells in tumors. Moreover, CCSC vaccine markedly increased the cytotoxicity of NK cells and the splenocytes, and promoted the release of IFN-γ, Perforin, and Granzyme B, and also reduced the TGF-ß1 expression. Additionally, CCSC vaccination enhanced the antibody production and decreased the myeloid derived suppressor cells and Treg subsets. More importantly, MUC1 knockdown partly impaired the anti-tumor efficacy of CCSC vaccine, whereas MUC1 overexpression dramatically enhanced the CCSC vaccine immunity. Overall, these results reveal a novel role and molecular mechanisms of MUC1 in CCSC vaccine against colorectal cancer.