An improved approach to generate IL-15+/+/TGFßR2-/- iPSC-derived natural killer cells using TALEN.

We present a TALEN-based workflow to generate and maintain dual-edited (IL-15+/+/TGFßR2-/-) iPSCs that produce enhanced iPSC-derived natural killer (iNK) cells for cancer immunotherapy. It involves using a cell lineage promoter for knocking in (KI) gene(s) to minimize the potential effects of expression of any exogenous genes on iPSCs. As a proof-of-principle, we KI IL-15 under the endogenous B2M promoter and show that it results in high expression of the sIL-15 in iNK cells but minimal expression in iPSCs. Furthermore, given that it is known that knockout (KO) of TGFßR2 in immune cells can enhance resistance to the suppressive TGF-ß signaling in the tumor microenvironment, we develop a customized medium containing Nodal that can maintain the pluripotency of iPSCs with TGFßR2 KO, enabling banking of these iPSC clones. Ultimately, we show that the dual-edited IL-15+/+/TGFßR2-/- iPSCs can be efficiently differentiated into NK cells that show enhanced autonomous growth and are resistant to the suppressive TGF-ß signaling.

Improved method for detecting protein-protein interactions using proximity ligation assay.

Proximity ligation assay has been widely used to detect protein-protein interaction in cells and tissues. While with great sensitivity, its specificity was often neglected. Here, we report the existence of varying levels of false positives observed with this assay and provide suggestions to minimize false positives for more accurate detection of protein-protein interactions, especially for membrane proteins.

Mei5-Sae3 stabilizes Dmc1 nucleating clusters for efficient Dmc1 assembly on RPA-coated single-stranded DNA.

Interhomolog recombination in meiosis requires a meiosis-specific recombinase, Dmc1. In Saccharomyces cerevisiae, the Mei5-Sae3 complex facilitates the loading of Dmc1 onto the replication protein A (RPA)-coated single-stranded DNA (ssDNA) to form nucleoprotein filaments. In vivo, Dmc1 and Mei5-Sae3 are interdependent in their colocalization on the chromosomes. However, the mechanistic role of Mei5-Sae3 in mediating Dmc1 activity remains unclear. We used single-molecule fluorescence resonance energy transfer and colocalization single-molecule spectroscopy experiments to elucidate how Mei5-Sae3 stimulates Dmc1 assembly on ssDNA and RPA-coated ssDNA. We showed that Mei5-Sae3 stabilized Dmc1 nucleating clusters with two to three molecules on naked DNA by preferentially reducing Dmc1 dissociation rates. Mei5-Sae3 also stimulated Dmc1 assembly on RPA-coated DNA. Using green fluorescent protein-labeled RPA, we showed the coexistence of an intermediate with Dmc1 and RPA on ssDNA before RPA dissociation. Moreover, the displacement efficiency of RPA depended on Dmc1 concentration, and its dependence was positively correlated with the stability of Dmc1 clusters on short ssDNA. These findings suggest a molecular model that Mei5-Sae3 mediates Dmc1 binding on RPA-coated ssDNA by stabilizing Dmc1 nucleating clusters, thus altering RPA dynamics on DNA to promote RPA dissociation.

Fitness implications of low-temperature storage for Eocanthecona furcellata (Hemiptera: Pentatomidae).

Exploring the impact of low-temperature storage on the fitness of natural enemy insects is crucial for practical field applications because this parameter directly influences their potential for population growth and effective pest control. Eocanthecona furcellata (Wolff) (Hemiptera: Pentatomidae) is widely used in biological pest control. This study aimed to identify optimal storage stages, temperatures, and durations for E. furcellata to produce high-quality individuals for practical use. The quality of E. furcellata after storage was evaluated by assessing parameters such as predatory capacity and fecundity, along with age-stage, two-sex life table. The findings revealed that the adult stage was the optimal storage form for E. furcellata, and the most favorable temperature for storage was 12 °C. Adult females had the highest predatory ability after 15 days of storage at 12 °C. Although survival rates declined with prolonged storage, they remained above 50% after 30 days, and longevity, fecundity, and predatory capacity of surviving individuals remained comparable to those of individuals in the control group (rearing at a constant temperature of 26 °C without low-temperature storage). The effects of low-temperature storage extended to the F1 generation of E. furcellata, which exhibited maximum mean longevity, fecundity, net reproductive rate, and mean generation time as well as fastest population growth after 30 days of storage at 12 °C. These results can be used to achieve optimal low-temperature storage conditions for E. furcellata production, particularly for extending its shelf life.

Stigmatization and Preferences in Monkeypox Vaccine Regimens.

A significant monkeypox (mpox) outbreak occurred in 2022, particularly involving sexual and gender minority (SGM) groups. Stigma and misperceptions have led to fear of being labeled a member of the SGM group when obtaining immunization for mpox. We hypothesized that the most recommended injection site, intradermal injection in the forearm, stresses stigmatization. We conducted an online survey in a medical center in Taiwan between May 2023 and June 2023 among adults who were going to receive the second preexposure vaccination. The questionnaire comprised questions about physical and psychological impacts of the first mpox vaccination and the preference for the second vaccination location and factors influencing the preference. A total of 2,827 individuals (98.4% male) completed the questionnaires. Intradermal injection in the forearm was related to greater physical and psychological impacts of local adverse events, especially discoloration. "Beauty," "scar," and "others' view" were the most significant factors influencing preference for vaccination regimens. Compared to intradermal injection in the forearm, subjects who cared about "others' views" were likely to prefer vaccination in the deltoid. The odds ratio for preferring intradermally injection in the deltoid over in the forearm was 1.88 (95% CI 1.38-2.56). The odds ratio for preferring subcutaneous injection in the deltoid over intradermally injection in the forearm was 1.69 (95% CI 1.23-2.32). The odds ratio for preferring intradermally injection in the deltoid regardless of the route over intradermally injection in the forearm was 2.11 (95% CI 1.53-2.92). This study demonstrated the adverse events of different mpox vaccination regimens and their association with stigma. Recognizing the factors affecting the preference for mpox vaccine regimens is crucial for easing the mental stress of vaccinee.

A novel tumor theranostic strategy based on metabolic glycoengineering and disulfidptosis.

Traditional metabolic glycoengineering involves participation of a monofunctional unnatural monosaccharide. To broaden the application boundary of this powerful technique, a bifunctional molecule, Ac4ManNSSN3, is designed and applied for a tumor theranostic strategy in this work. The results from both cell and animal experiments show good in situ tumor detection and tumor inhibition effects.

Actively Targeting Redox-Responsive Multifunctional Micelles for Synergistic Chemotherapy of Cancer.

Stimuli-responsive polymeric micelles decorated with cancer biomarkers represent an optimal choice for drug delivery applications due to their ability to enhance therapeutic efficacy while mitigating adverse side effects. Accordingly, we synthesized a digoxin-modified novel multifunctional redox-responsive disulfide-linked poly(ethylene glycol-b-poly(lactic-co-glycolic acid) copolymer (Bi(Dig-PEG-PLGA)-S2) for the targeted and controlled release of doxorubicin (DOX) in cancer cells. Within the micellar aggregate, the disulfide bond confers redox responsiveness, while the presence of the digoxin moiety acts as a targeting agent and chemosensitizer for DOX. Upon self-assembly in aqueous solution, Bi(Dig-PEG-PLGA)-S2 formed uniformly distributed spherical micelles with a hydrodynamic diameter (D h ) of 58.36 ± 0.78 nm and a zeta potential of -24.71 ± 1.01 mV. The micelles exhibited desirable serum and colloidal stability with a substantial drug loading capacity (DLC) of 6.26% and an encapsulation efficiency (EE) of 83.23%. In addition, the release of DOX demonstrated the redox-responsive behavior of the micelles, with approximately 89.41 ± 6.09 and 79.64 ± 6.68% of DOX diffusing from DOX@Bi(Dig-PEG-PLGA)-S2 in the presence of 10 mM GSH and 0.1 mM H2O2, respectively, over 96 h. Therefore, in HeLa cell lines, DOX@Bi(Dig-PEG-PLGA)-S2 showed enhanced intracellular accumulation and subsequent apoptotic effects, attributed to the targeting ability and chemosensitization potential of digoxin. Hence, these findings underscore the promising characteristics of Bi(Dig-PEG-PLGA)-S2 as a multifunctional drug delivery vehicle for cancer treatment.

Surgical outcomes and intraoperative parameters of 3D visualization system versus conventional microscopes for macular surgery in highly myopic eyes, a prospective randomized clinical trial.

PURPOSE: To investigate the surgical outcomes and intraoperative parameters of 3D visualization system for macular diseases in highly myopic eyes. METHODS: In this single-center, prospective, randomized, comparative interventional study, 40 highly myopic eyes (axial length > 26mm) were randomly assigned to either a 3D visualization system or a conventional microscope (CM) group. Surgical outcomes and intraoperative parameters, including the number of indocyanine green (ICG) injections, surgical time, and epiretinal membrane (ERM)/ internal limiting membrane (ILM) peeling time, were compared. RESULTS: The 3D group required significantly fewer ICG injections (1.3 ± 0.5 vs. 2.3 ± 0.7, p < 0.001), had shorter ERM/ILM peeling times (522.8 ± 258.0 vs. 751.8 ± 320.2 sec, p < 0.05), and experienced fewer intraoperative retinal hemorrhages (0 vs. 7 cases, p < 0.05) compared to the CM group. Anatomical and functional outcomes were comparable between the two groups. CONCLUSION: The 3D system exhibited a lower number of ICG injections, shorter ERM/ILM peeling times and a reduced incidence of intraoperative retinal hemorrhages, suggesting the 3D visualization system may offer advantages for macular surgery in highly myopic eyes.

Stress-sensitive neural circuits change the gut microbiome via duodenal glands.

Negative psychological states impact immunity by altering the gut microbiome. However, the relationship between brain states and microbiome composition remains unclear. We show that Brunner's glands in the duodenum couple stress-sensitive brain circuits to bacterial homeostasis. Brunner's glands mediated the enrichment of gut Lactobacillus species in response to vagus nerve stimulation. Cell-specific ablation of the glands markedly suppressed Lactobacilli counts and heightened vulnerability to infection. In the forebrain, we mapped a vagally mediated, polysynaptic circuit connecting the central nucleus of the amygdala to Brunner's glands. Chronic stress suppressed central amygdala activity and phenocopied the effects of gland lesions. Conversely, excitation of either the central amygdala or parasympathetic vagal neurons activated Brunner's glands and reversed the effects of stress on the gut microbiome and immunity. The findings revealed a tractable brain-body mechanism linking psychological states to host defense.

APOE ε4-associated downregulation of the IL-7/IL-7R pathway in effector memory T cells: Implications for Alzheimer's disease.

INTRODUCTION: The apolipoprotein E (APOE) ε4 allele exerts a significant influence on peripheral inflammation and neuroinflammation, yet the underlying mechanisms remain elusive. METHODS: The present study enrolled 54 patients diagnosed with late-onset Alzheimer's disease (AD; including 28 APOE ε4 carriers and 26 non-carriers). Plasma inflammatory cytokine concentration was assessed, alongside bulk RNA sequencing (RNA-seq) and single-cell RNA sequencing (scRNA-seq) analysis of peripheral blood mononuclear cells (PBMCs). RESULTS: Plasma tumor necrosis factor α, interferon γ, and interleukin (IL)-33 levels increased in the APOE ε4 carriers but IL-7 expression notably decreased. A negative correlation was observed between plasma IL-7 level and the hippocampal atrophy degree. Additionally, the expression of IL-7R and CD28 also decreased in PBMCs of APOE ε4 carriers. ScRNA-seq data results indicated that the changes were mainly related to the CD4+ Tem (effector memory) and CD8+ Tem T cells. DISCUSSION: These findings shed light on the role of the downregulated IL-7/IL-7R pathway associated with the APOE ε4 allele in modulating neuroinflammation and hippocampal atrophy. HIGHLIGHTS: The apolipoprotein E (APOE) ε4 allele decreases plasma interleukin (IL)-7 and aggravates hippocampal atrophy in Alzheimer's disease. Plasma IL-7 level is negatively associated with the degree of hippocampal atrophy. The expression of IL-7R signaling decreased in peripheral blood mononuclear cells of APOE ε4 carriers Dysregulation of the IL-7/IL-7R signal pathways enriches T cells.

Genomic and pathogenicity analysis of two novel highly pathogenic recombinant NADC30-like PRRSV strains in China, in 2023.

Porcine reproductive and respiratory syndrome viruses (PRRSVs) exhibit high mutability and recombination, posing challenges to their immunization and control. This study isolated two new PRRSV strains, GD-7 and GX-3, from samples collected in Guangdong and Guangxi in 2023. Whole-genome sequencing, along with phylogenetic and recombination analyses, confirmed that GD-7 and GX-3 are natural novel recombinant strains of NADC30 PRRSV. Moreover, we established a pathogenicity model for piglets and sows based on the two isolates. The results of piglet pathogenicity revealed that both GD-7 and GX-3 caused clinical symptoms such as fever, loss of appetite, depression, and slow weight gain. Moreover, we observed that the mortality rate of GD-7-inoculated group piglets was 33.3%, which was similar to that of piglets infected with other highly pathogenic PRRSV strains and exceeded the mortality rate of most NADC30-like PRRSV. In pregnant sow models, the survival rate of sows in the GD-7 group was 75%, in contrast to the GX-3 group, where no sow mortality was observed, and both strains resulted in abortion, mummified fetuses, and stillbirths. These results highlight the elevated pathogenicity of these recombinant strains in sows, with GD-7 mainly causing sows to abort, and GX-3 mainly causing sows to give birth to mummified fetuses. This study introduces two distinct clinical recombinant PRRSV strains that differ from the prevalent strains in China. This research furthers our understanding of the epidemiology of PRRSV and underscores the significance of ongoing monitoring and research in the face of evolving virus strains. Moreover, these discoveries act as early warnings, underscoring the necessity for active control and immunization against PRRSV.IMPORTANCESince the discovery of NADC30-like PRRSV in China in 2013, it has gradually become the dominant strain of PRRSV in China. NADC30-like PRRSV exhibits high recombination characteristics, constantly recombining with different strains, leading to the emergence of numerous novel strains. Of particular importance is the observation that NADC30-like PRRSV with different recombination patterns exhibits varying pathogenicity, which has a significant impact on the pig farming industry. This emphasizes the necessity of monitoring and responding to evolving PRRSV strains to develop effective immunization and control strategies. In this paper, we conducted pathogenicity studies on the isolated NADC30-like PRRSV and analyzed the differences in the genomes and pathogenicity of the different strains by recording clinical symptoms, temperature changes, detoxification tests, and changes in viremia and histopathology in infected pigs. This was done to provide a theoretical basis for the epidemiological situation and epidemic prevention and control of PRRSV.

Potential damage to ovarian reserve from laparoscopic electrocoagulation in endometriomas and benign ovarian cysts: a systematic review and meta-analysis.

PURPOSE: Laparoscopic cystectomy for ovarian endometriomas and benign ovarian cysts is often conducted through hemostatic methods, with bipolar electrocoagulation as a common approach. This study evaluated the impact of electrocoagulation, primarily through bipolar energy, versus nonthermal hemostatic methods on ovarian reserve in patients undergoing laparoscopic cystectomy for ovarian endometriomas and benign ovarian cysts. METHODS: A systematic review with meta-analysis was conducted by searching the Cochrane Library, PubMed, EMBASE, and Web of Science databases. Randomized controlled trials (RCTs) comparing the impact of nonthermal hemostatic methods and electrocoagulation on the ovarian reserve during laparoscopic cystectomy were included. The Cochrane Risk of Bias Tool for Randomized Controlled Trials (ROB 2.0) was utilized to assess the quality of the included studies. The meta-analysis included 13 RCTs involving 1043 patients. Postoperative serum anti-Müllerian hormone (AMH) levels and antral follicle counts (AFCs) were analyzed using Review Manager ver. 5.4. RESULTS: Compared with the bipolar group, patients with endometriomas in the nonthermal hemostatic group exhibited significantly higher postoperative AMH levels at 1, 3, 6, and 12 months. Conversely, no significant differences in AMH levels were observed in patients with benign ovarian cysts. Similarly, AFCs showed no significant differences, except for lower postoperative AFCs in patients with endometrioma in the electrocoagulation group. CONCLUSION: Nonthermal hemostatic methods are associated with more effective preservation of the ovarian reserve compared with bipolar electrocoagulation in laparoscopic cystectomy for ovarian endometriomas. However, no significant impact of bipolar electrocoagulation on the ovarian reserve was observed in patients with benign ovarian cysts. TRIAL REGISTRATION: Registered in PROSPERO on April 10, 2023; ID # CRD42023413158.

Relationship between genotype, phenotype, and refractive status in patients of inherited retinal degeneration.

BACKGROUND: To elucidate the relationship between inherited retinal disease, visual acuity and refractive error development in Asian patients. SUBJECTS: Five hundred phakic eyes with refractive data were analysed in this retrospective cohort. Diseases were categorized by clinical phenotypes, and the prevalent genotypes identified in the Taiwan Inherited Retinal Degeneration Project were analysed. Consecutive surveys in Taiwan have provided the rates of myopia in the general population. RESULTS: No differences were observed among the disease phenotypes with respect to myopia (P = 0.098) and high myopia rates (P = 0.037). The comparison of refractive error between retinitis pigmentosa and diseases mainly affecting the central retina showed no difference, and the refraction analyses in diseases of different onset ages yielded no significance. Moreover, there was no difference in the myopia rate between the diseases and general population. Among the genotypes, a higher spherical equivalent was seen in RPGR and PROM1-related patients and emmetropic trends were observed in patients with CRB1 and PRPF31 mutations. Furthermore, significantly poorer visual acuity was found in ABCA4, CRB1 and PROM1-related patients, and more preserved visual acuity was seen in patients with EYS, USH2A, and RDH12 mutations. CONCLUSIONS: No significant differences were observed in visual acuity, refractive state and myopia rate between patients with inherited retinal disease and the general population, and different subtypes of inherited retinal disease shared similar refractive state, except for higher cylindrical dioptres found in patients with Leber's congenital amaurosis. The heterogeneity of disease-causing genes in Asian patients may lead to variable refractive state.

Laser-Induced Interference to Infrared Detector Using Continuous Wave and Short-Pulse Lasers.

The response of a DPbS3200 infrared detector irradiated by a nanosecond pulsed laser and CW laser has been investigated to study laser-induced interference. A laser interference experiment system was constructed to measure the time-varying response signal. A nanosecond pulsed laser and a CW laser of 10 Hz were used, with a 1064 nm wavelength and a millimeter-scale irradiation spot diameter. Firstly, the characteristics of transient interference signals induced by pulsed lasers were analyzed. Then, the characteristics of response signal interference by both CW laser and pulsed laser irradiation were further investigated. The results showed that the pulsed laser only produced transient interference. However, the CW laser led to a significant amplitude reduction of the response signal, which could continuously interfere in the operating time. For transient interferences, the amplitude of the interference signal increased linearly with the laser fluence. The relation between the pulse repetition rate of the incident laser and the operating frequency of the detector determined the numbers of transient interference signals in one response period; for the interference induced by both the CW laser and pulsed laser, CW laser interference played a leading role when CW laser power density increased to 4.1 W/cm2 or more. As the CW laser fluence reached 6.1 W/cm2, the PbS infrared detector was no longer able to detect any signal, which caused temporary blindness. In the end, a probit model was used to determine the interference threshold.

Damage Characteristics Analysis of Laser Ablation Triple-Junction Solar Cells Based on Electroluminescence Characteristics.

To study the physical property effects of the laser on GaInP/GaAs/Ge solar cells and their sub-cell layers, a pulsed laser with a wavelength of 532 nm was used to irradiate the solar cells under various energy conditions. The working performance of the cell was measured with a source meter. The electroluminescence (EL) characteristics were assessed using an ordinary and an infrared camera. Based on the detailed balance theory, in the voltage characteristics of an ideal pristine cell, the GaInP layer made the most significant voltage contribution, followed by the GaAs layer, with the Ge layer contributing the least. When a bias voltage was applied to the pristine cell, the top GaInP cell emitted red light at 670 nm, the middle GaAs cell emitted near-infrared light at 926 nm, and the bottom Ge cell emitted infrared light at 1852 nm. In the experiment, the 532 nm laser wavelength within the response spectrum bands of the GaInP layer and the laser passed through the glass cover slip and directly interacted with the GaInP layer. The experimental results indicated that the GaInP layer first exhibited different degrees of damage under laser irradiation, and the cell voltage was substantially attenuated. The GaInP/GaAs/Ge solar cell showed a decrease in electrical and light emission characteristics. As the laser energy increased, the cell's damage intensified, gradually leading to a loss of photoelectric conversion capability, the near-complete disappearance of red light emission, and a gradual degradation of near-infrared emission properties. The EL imaging revealed varying damage states across the triple-junction gallium arsenide solar cell's sub-cells.

Assessment of foetal dose and occupational exposure for pregnant workers in nuclear medicine using the Taiwanese pregnancy phantom.

For pregnant workers in nuclear medicine, radiation doses can pose a risk to their foetus. However, foetal radiation doses cannot be measured directly. In this study, a method of estimating foetal radiation doses was developed through simulations and measurements of phantoms of pregnant women in the three trimesters. The uterus and abdominal surface doses for monoenergetic photons (137Cs) and medical diagnostic X-rays were measured, and uterine dose conversion coefficients (UDCCs) were calculated. The accuracy of the UDCC estimates were validated for measurements from thermoluminescent dosemeter (TLD) chips and TLD badges on the abdomen or chest. The foetal effective dose could be estimated using TLD chips and TLD badges on the abdomen or chest, or through literature estimation method. The proposed method can be used to easily and accurately estimate foetal effective doses from chest-worn TLD badges, ensuring accurate estimation in the early stage of pregnancy when a worker may not yet be wearing an abdominal badge. A flowchart for applying the UDCC method to approximate a foetal dose is also provided to ensure that total doses remain below the maximum of 1 mSv recommended in the International Commission on Radiological Protection 103 guidelines.

Remote Sensing Change Detection With Bitemporal and Differential Feature Interactive Perception.

Recently, the transformer has achieved notable success in remote sensing (RS) change detection (CD). Its outstanding long-distance modeling ability can effectively recognize the change of interest (CoI). However, in order to obtain the precise pixel-level change regions, many methods directly integrate the stacked transformer blocks into the UNet-style structure, which causes the high computation costs. Besides, the existing methods generally consider bitemporal or differential features separately, which makes the utilization of ground semantic information still insufficient. In this paper, we propose the multiscale dual-space interactive perception network (MDIPNet) to fill these two gaps. On the one hand, we simplify the stacked multi-head transformer blocks into the single-layer single-head attention module and further introduce the lightweight parallel fusion module (LPFM) to perform the efficient information integration. On the other hand, based on the simplified attention mechanism, we propose the cross-space perception module (CSPM) to connect the bitemporal and differential feature spaces, which can help our model suppress the pseudo changes and mine the more abundant semantic consistency of CoI. Extensive experiment results on three challenging datasets and one urban expansion scene indicate that compared with the mainstream CD methods, our MDIPNet obtains the state-of-the-art (SOTA) performance while further controlling the computation costs.

Huangqin tang alleviates colitis-associated colorectal cancer via amino acids homeostasisand PI3K/AKT/mtor pathway modulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Huangqin Tang (HQT), a traditional Chinese medicine formula, is commonly used in clinical practice for the treatment of inflammatory bowel diseases. It has been reported that HQT exerts antitumor effects on colitis-associated colorectal cancer (CAC). However, the mechanism by which HQT interferes with the inflammation-to-cancer transformation remains unclear. AIMS OF THE STUDY: The purpose of this study was to dynamically evaluate the efficacy of HQT in alleviating or delaying CAC and to reveal the underlying mechanism. METHODS: We established a mouse model of CAC using azoxymethane combined with 1.5% dextran sodium sulphate. The efficacy of HQT was evaluated based on pathological sections and serum biochemical indices. Subsequently, amino acids (AAs) metabolism analyses were performed using ultra-performance liquid chromatography-tandem mass spectrometry, and the phosphatidylinositol 3 kinase/protein kinase B/mechanistic target of rapamycin (PI3K/AKT/mTOR) pathway was detected by western blotting. RESULTS: The data demonstrated that HQT could alleviate the development of CAC in the animal model. HQT effectively reduced the inflammatory response, particularly interleukin-6 (IL-6), in the inflammation induction stage, as well as in the stages of proliferation initiation and tumorigenesis. During the proliferation initiation and tumorigenesis stages, immunohistochemistry staining showed that the expression of the proliferation marker Ki67 was reduced, while apoptosis was increased in the HQT group. Accordingly, HQT substantially decreased the levels of specific AAs in the colon with CAC, including glutamic acid, glutamine, arginine, and isoleucine. Furthermore, HQT significantly inhibited the activated PI3K/AKT/mTOR pathway, which may contribute to suppression of cell proliferation and enhancement of apoptosis. CONCLUSION: HQT is effective in alleviating and delaying the colon "inflammation-to-cancer". The mechanism of action may involve HQT maintained AAs metabolism homeostasis and regulated PI3K/AKT/mTOR pathway, so as to maintain the balance between proliferation and apoptosis, and then interfere in the occurrence and development of CAC.

Colonization compatibility with Bacillus altitudinis confers soybean seed rot resistance.

The plant microbiome and plant-associated bacteria are known to support plant health, but there are limited studies on seed and seedling microbiome to reveal how seed-associated bacteria may confer disease resistance. In this study, the application of antibiotics on soybean seedlings indicated that seed-associated bacteria were involved in the seed rot resistance against a soil-borne pathogen Calonectria ilicicola, but this resistance cannot be carried to withstand root rot. Using PacBio 16S rRNA gene full-length sequencing and microbiome analyses, 14 amplicon sequence variants (ASVs) including 2 ASVs matching to Bacillus altitudinis were found to be more abundant in the four most resistant varieties versus the four most susceptible varieties. Culture-dependent isolation obtained two B. altitudinis isolates that both exhibit antagonistic capability against six fungal pathogens. Application of B. altitudinis on the most resistant and susceptible soybean varieties revealed different colonization compatibility, and the seed rot resistance was restored in the five varieties showing higher bacterial colonization. Moreover, quantitative PCR confirmed the persistence of B. altitudinis on apical shoots till 21 days post-inoculation (dpi), but 9 dpi on roots of the resistant variety TN5. As for the susceptible variety HC, the persistence of B. altitudinis was only detected before 6 dpi on both shoots and roots. The short-term colonization of B. altitudinis on roots may explain the absence of root rot resistance. Collectively, this study advances the insight of B. altitudinis conferring soybean seed rot resistance and highlights the importance of considering bacterial compatibility with plant varieties and colonization persistence on plant tissues.

Separable nanocomposite hydrogel microneedles for intradermal and sustained delivery of antigens to enhance adaptive immune responses.

Vaccines play a critical role in combating infectious diseases and cancers, yet improving their efficacy remains challenging. Here, we introduce a separable nanocomposite hydrogel microneedle (NHMN) patch designed for intradermal and sustained delivery of ovalbumin (OVA), a model antigen, to enhance adaptive immune responses. The NHMN patch consists of an array of OVA-loaded microneedles made from photo-cross-linked methacrylated hyaluronic acid and laponite (LAP), supported by a hyaluronic acid backing. The incorporation of LAP not only enhances the mechanical strength of the pure hydrogel microneedles but also significantly prolongs OVA release. Furthermore, in vitro cell experiments demonstrate that NHMNs effectively activate dendritic cells without compromising cell viability. Upon skin penetration, NHMNs detach from the backing as the hyaluronic acid rapidly dissolves upon contact with the skin interstitial fluid, thereby acting as antigen reservoirs to release antigens to abundant skin dendritic cells. NHMNs containing 0.5% w/v LAP achieved a 15-day OVA release in vivo. Immunization studies demonstrate that the intradermal and sustained release of OVA via NHMNs elicited stronger and longer-lasting adaptive immune responses compared to conventional bolus injection. Given its easy to use, painless and minimally invasive features, the NHMN patch shows promise in improving vaccination accessibility and efficacy against a range of diseases. STATEMENT OF SIGNIFICANCE: The study introduces a separable nanocomposite hydrogel microneedle (NHMN) patch. This patch consists of an array of ovalbumin (OVA, a model antigen)-loaded microneedles made from photo-cross-linked methacrylated hyaluronic acid and laponite, with a hyaluronic acid backing, designed for intradermal and sustained delivery of antigens. This patch addresses several key challenges in traditional vaccination methods, including poor antigen uptake and presentation, and rapid systematic clearance. The incorporation of laponite enhances mechanical strength of microneedles, promotes dendritic cell activation, and significantly slows down antigen release. NHMN-based vaccination elicits stronger and longer-lasting adaptive immune responses compared to conventional bolus injection. This NHMN patch holds great potential for improving the efficacy, accessibility, and patient comfort of vaccinations against a range of diseases.