Freeze-Casting of Alumina and Permeability Analysis Based on a 3D Microstructure Reconstructed Using Generative Adversarial Networks.

In this study, alumina ceramics with hierarchical pores were successfully fabricated using freeze casting. Experimental studies show that both the solid loading of the slurry and the thermal insulation layer at the interface of the slurry and cooling plate can influence the pore characteristics of cast samples. In order to examine the pore characteristics and evaluate the permeability of the freeze-cast samples fabricated under different conditions, a generative adversarial network (GAN) method was employed to reconstruct the three-dimensional (3D) microstructure from two-dimensional (2D) scanning electron microscopy (SEM) images of the samples. Furthermore, GAN 3D reconstruction was validated against X-ray tomography 3D reconstruction results. Based on the GAN reconstructed microstructures, the permeability and pore distribution of the various samples were analyzed. The sample cast with 35 wt.% solid loading shows an optimal permeability.

Efficient expansion and CRISPR-Cas9-mediated gene correction of patient-derived hepatocytes for treatment of inherited liver diseases.

Cell-based ex vivo gene therapy in solid organs, especially the liver, has proven technically challenging. Here, we report a feasible strategy for the clinical application of hepatocyte therapy. We first generated high-quality autologous hepatocytes through the large-scale expansion of patient-derived hepatocytes. Moreover, the proliferating patient-derived hepatocytes, together with the AAV2.7m8 variant identified through screening, enabled CRISPR-Cas9-mediated targeted integration efficiently, achieving functional correction of pathogenic mutations in FAH or OTC. Importantly, these edited hepatocytes repopulated the injured mouse liver at high repopulation levels and underwent maturation, successfully treating mice with tyrosinemia following transplantation. Our study combines ex vivo large-scale cell expansion and gene editing in patient-derived transplantable hepatocytes, which holds potential for treating human liver diseases.

Effect of Phenol and Alkylamide Interaction on α-Glucosidase Inhibition and Cellular Antioxidant Activity during In Vitro Digestion: Using Szechuan Pepper (Zanthoxylum genus) as a Model.

Although recent evidence indicated significant phenol and alkylamide interaction in aqueous solutions, the gastrointestinal digestion influence of the combination remains unclear. This study aims to investigate phenol and alkylamide interaction during in vitro digestion, focusing on bioaccessibility and bioactivity, including α-glucosidase inhibition and cellular antioxidant activity. Additionally, the structural mechanism of phenol and alkylamide interaction during in vitro digestion was explored. The results indicated that the presence of phenols and alkylamides significantly increased or decreased their respective bioaccessibility, depending on the Zanthoxylum varieties. Furthermore, although antagonistic phenol/alkylamide interaction was evident during α-glucosidase inhibition, cellular oxidative stress alleviation, and antioxidant gene transcription upregulation, this effect weakened gradually as digestion progressed. Glycoside bond cleavage and the methylation of phenols as well as alkylamide isomerization and addition were observed during digestion, modifying the hydrogen bonding sites and interaction behavior. This study provided insights into the phenol/alkylamide interaction in the gastrointestinal tract.

Fork coupling directs DNA replication elongation and termination.

DNA replication is initiated at multiple loci to ensure timely duplication of eukaryotic genomes. Sister replication forks progress bidirectionally, and replication terminates when two convergent forks encounter one another. To investigate the coordination of replication forks, we developed a replication-associated in situ HiC method to capture chromatin interactions involving nascent DNA. We identify more than 2000 fountain-like structures of chromatin contacts in human and mouse genomes, indicative of coupling of DNA replication forks. Replication fork interaction not only occurs between sister forks but also involves forks from two distinct origins to predetermine replication termination. Termination-associated chromatin fountains are sensitive to replication stress and lead to coupled forks-associated genomic deletions in cancers. These findings reveal the spatial organization of DNA replication forks within the chromatin context.

Classification of Camellia oleifera Diseases in Complex Environments by Attention and Multi-Dimensional Feature Fusion Neural Network.

The use of neural networks for plant disease identification is a hot topic of current research. However, unlike the classification of ordinary objects, the features of plant diseases frequently vary, resulting in substantial intra-class variation; in addition, the complex environmental noise makes it more challenging for the model to categorize the diseases. In this paper, an attention and multidimensional feature fusion neural network (AMDFNet) is proposed for Camellia oleifera disease classification network based on multidimensional feature fusion and attentional mechanism, which improves the classification ability of the model by fusing features to each layer of the Inception structure and enhancing the fused features with attentional enhancement. The model was compared with the classical convolutional neural networks GoogLeNet, Inception V3, ResNet50, and DenseNet121 and the latest disease image classification network DICNN in a self-built camellia disease dataset. The experimental results show that the recognition accuracy of the new model reaches 86.78% under the same experimental conditions, which is 2.3% higher than that of GoogLeNet with a simple Inception structure, and the number of parameters is reduced to one-fourth compared to large models such as ResNet50. The method proposed in this paper can be run on mobile with higher identification accuracy and a smaller model parameter number.

Cohesin maintains replication timing to suppress DNA damage on cancer genes.

Cohesin loss-of-function mutations are frequently observed in tumors, but the mechanism underlying its role in tumorigenesis is unclear. Here, we found that depletion of RAD21, a core subunit of cohesin, leads to massive genome-wide DNA breaks and 147 translocation hotspot genes, co-mutated with cohesin in multiple cancers. Increased DNA damages are independent of RAD21-loss-induced transcription alteration and loop anchor disruption. However, damage-induced chromosomal translocations coincide with the asymmetrically distributed Okazaki fragments of DNA replication, suggesting that RAD21 depletion causes replication stresses evidenced by the slower replication speed and increased stalled forks. Mechanistically, approximately 30% of the human genome exhibits an earlier replication timing after RAD21 depletion, caused by the early initiation of >900 extra dormant origins. Correspondingly, most translocation hotspot genes lie in timing-altered regions. Therefore, we conclude that cohesin dysfunction causes replication stresses induced by excessive DNA replication initiation, resulting in gross DNA damages that may promote tumorigenesis.

Assessment of genetic variants in D2 dopamine receptor (DRD2) gene as risk factors for post-traumatic stress disorder (PTSD) and major depressive disorder (MDD): A systematic review and meta-analysis.

BACKGROUND: Many studies have performed assessments of genetic variants in the D2 dopamine receptor (DRD2) gene as risk factors for post-traumatic stress disorder (PTSD) and major depressive disorder (MDD). However, the results are inconsistent. This meta-analysis aimed to systematically summarize published data to evaluate the reliable association between the DRD2 genetic variants and the risk of PTSD and MDD. METHODS: A systematic literature search was conducted using the Web of Science, PubMed, Google Scholar, Excerpta Medica Database (EMBASE), Springer, ScienceDirect, Wiley Online Library, Cochrane Central Register of Controlled Trials, Chinese Biomedical Literature Database (CBM), WANFANG Data, CQVIP, and Chinese National Knowledge Infrastructure (CNKI) databases before January 1st, 2022. RESULTS: A total of 27 genetic variants in the DRD2 gene were retrieved, and 7 of them met the inclusion criteria for meta-analysis. Our meta-analysis results indicated that the rs1800497 (TaqIA) polymorphism was significantly associated with the increased risk of PTSD (Dominant model (A1A1 + A1A2 vs. A2A2): OR = 1.49, 95 % CI, 1.08-2.04 Z = 2.46, P = 0.014). Subgroup analysis for ethnicity suggested that a significantly increased risk of PTSD was observed in Asians (Dominant model (A1A1 + A1A2 vs. A2A2): OR = 1.39, 95 % CI, 1.08-1.79, Z = 2.60, P = 0.009) and Caucasians (Dominant model (A1A1 + A1A2 vs. A2A2): OR = 1.87, 95 % CI 1.02-3.41, Z = 2.04, P = 0.042). Meanwhile, we detected significant association strengths between the rs1799978 and rs2075652 polymorphisms in the DRD2 gene and MDD (for rs1799978, Homozygote comparison (GG vs. AA): OR = 0.60, 95 % CI = 0.37-0.97, Z = 2.08, P = 0.038; for rs2075652, Homozygote comparison (AA vs. GG): OR = 1.82, 95 % CI = 1.32-2.50, Z = 3.67, P < 0.001). Our cumulative meta-analyses indicated a continuous trend toward association strength with PTSD and MDD. CONCLUSIONS: This meta-analysis indicated that genetic variants in the DRD2 gene might potentially contribute to genetic susceptibility for PTSD and MDD. The utilization of DRD2 genetic variants as risk factors for PTSD and MDD requires further validation by large well-designed case-control studies.

Understanding the Configurational Entropy Evolution in Metal-Phosphorus Solid Solution for Highly Reversible Li-Ion Batteries.

The high-entropy materials (HEM) have attracted increasing attention in catalysis and energy storage due to their large configurational entropy and multiunique properties. However, it is failed in alloying-type anode due to their Li-inactive transition-metal compositions. Herein, inspired by high-entropy concept, the Li-active elements instead of transition-metal ones are introduced for metal-phosphorus synthesis. Interestingly, a new Znx Gey Cuz Siw P2 solid solution is successfully synthesized as proof of concept, which is first verified to cubic system in F-43m. More specially, such Znx Gey Cuz Siw P2 possesses wide-range tunable region from 9911 to 4466, in which the Zn0.5 Ge0.5 Cu0.5 Si0.5 P2 accounts for the highest configurational entropy. When served as anode, Znx Gey Cuz Siw P2 delivers large capacity (>1500 mAh g-1 ) and suitable plateau (≈0.5 V) for energy storage, breaking the conventional view that HEM is helpless for alloying anode due to its transition-metal compositions. Among them, the Zn0.5 Ge0.5 Cu0.5 Si0.5 P2 exhibits the highest initial coulombic efficiency (ICE) (93%), Li-diffusivity (1.11 × 10-10 ), lowest volume-expansion (34.5%), and best rate performances (551 mAh g-1 at 6400 mA g-1 ) owing to its largest configurational entropy. Possible mechanism reveals the high entropy stabilization enables good accommodation of volume change and fast electronic transportation, thus supporting superior cyclability and rate performances. This large configurational entropy strategy in metal-phosphorus solid solution may open new avenues to develop other high-entropy materials for advanced energy storage.

CRISPR/Cas9-induced structural variations expand in T lymphocytes in vivo.

CRISPR/Cas9 has been adapted to disrupt endogenous genes in adoptive T-lymphocyte therapy to prevent graft-versus-host disease. However, genome editing also generates prevalent deleterious structural variations (SVs), including chromosomal translocations and large deletions, raising safety concerns about reinfused T cells. Here, we dynamically monitored the progression of SVs in a mouse model of T-cell receptor (TCR)-transgenic T-cell adoptive transfer, mimicking TCR T therapeutics. Remarkably, CRISPR/Cas9-induced SVs persist and undergo clonal expansion in vivo after three weeks or even two months, evidenced by high enrichment and low junctional diversity of identified SVs post infusion. Specifically, we detected 128 expanded translocations, with 20 615 as the highest number of amplicons. The identified SVs are stochastically selected among different individuals and show an inconspicuous locus preference. Similar to SVs, viral DNA integrations are routinely detected in edited T cells and also undergo clonal expansion. The persistent SVs and viral DNA integrations in the infused T cells may constantly threaten genome integrity, drawing immediate attention to the safety of CRISPR/Cas9-engineered T cells mediated immunotherapy.

Seasonal expressions of GPR41 and GPR43 in the colon of the wild ground squirrels (Spermophilus dauricus).

G-protein-coupled receptor 41 (GPR41) and G-protein-coupled receptor 43 (GPR43) are important short-chain fatty acids (SCFAs) receptors. Previous studies indicated that GPR41 and GPR43 are involved in the secretion of gastrointestinal peptides, and glucose and lipid metabolism, and are closely related to obesity and type II diabetes, and other diseases. The purpose of the study was to explore the relationship between the GPR41 and GPR43 and seasonal breeding, and provide new prospects for further exploring the nutritional needs of breeding. We identified the localization and expression levels of GPR41 and GPR43 in the colon of the wild ground squirrels (Spermophilus dauricus) both in the breeding season and non-breeding season. The histological results revealed that the lumen diameter of the colon had obvious seasonal changes, and the diameter of the colonic lumen in the non-breeding season was larger than that in the breeding season. Immunohistochemical staining suggested GPR41 and GPR43 have expressed in the simple layer columnar epithelium. In addition, compared with the breeding season, the mRNA and protein expression levels of GPR41 and GPR43 in the colon were higher during the non-breeding season. In general, these results indicated GPR41 and GPR43 might play a certain role in regulating seasonal breeding.

Transcription shapes DNA replication initiation to preserve genome integrity.

BACKGROUND: Early DNA replication occurs within actively transcribed chromatin compartments in mammalian cells, raising the immediate question of how early DNA replication coordinates with transcription to avoid collisions and DNA damage. RESULTS: We develop a high-throughput nucleoside analog incorporation sequencing assay and identify thousands of early replication initiation zones in both mouse and human cells. The identified early replication initiation zones fall in open chromatin compartments and are mutually exclusive with transcription elongation. Of note, early replication initiation zones are mainly located in non-transcribed regions adjacent to transcribed regions. Mechanistically, we find that RNA polymerase II actively redistributes the chromatin-bound mini-chromosome maintenance complex (MCM), but not the origin recognition complex (ORC), to actively restrict early DNA replication initiation outside of transcribed regions. In support of this finding, we detect apparent MCM accumulation and DNA replication initiation in transcribed regions due to anchoring of nuclease-dead Cas9 at transcribed genes, which stalls RNA polymerase II. Finally, we find that the orchestration of early DNA replication initiation by transcription efficiently prevents gross DNA damage. CONCLUSION: RNA polymerase II redistributes MCM complexes, but not the ORC, to prevent early DNA replication from initiating within transcribed regions. This RNA polymerase II-driven MCM redistribution spatially separates transcription and early DNA replication events and avoids the transcription-replication initiation collision, thereby providing a critical regulatory mechanism to preserve genome stability.

Interactions between Phenols and Alkylamides of Sichuan Pepper (Zanthoxylum Genus) in α-Glucosidase Inhibition: A Structural Mechanism Analysis.

The phenols and alkylamides in 26 varieties of Zanthoxylum pericarps (ZP) were comparatively identified, and the contribution of these key components to the inhibition of in vitro α-glucosidase (α-Glu) was confirmed using principal component analysis (PCA) and ingredient recombination models. Additionally, spectrophotometric assays, nuclear magnetic resonance (NMR), and molecular docking (MD) were employed to characterize the interactions among key components in ZP when exposed to α-Glu. Four phenols and hydroxy-α-sanshool (α-SOH), which were recognized as main ingredients, presented an antagonistic effect in the inhibition of α-Glu. 1H NMR demonstrated chemical shifts of certain hydrogens in the B phenolic ring and tetraenyl group, indicating a possible p-π conjugation between phenols and α-SOH. In addition, using MD analysis, the phenol-binding sites were observed to be negatively affected when α-SOH initially interacted with α-Glu. The combined results of the NMR and MD clarified the structural mechanism behind phenol/α-SOH antagonistic behavior in α-Glu inhibition.

Fabricating efficient and stable quasi-3D and 3D/2D perovskite solar cells with 2D-sheets connected by inorganic type ionic-bond.

In this work, we report a novel two-dimensional (2D) (SrBr)2PbI4perovskite layered architecture, which were formed by the reaction of strontium bromide (SrBr2), strontium iodide (SrI2) and lead iodide (PbI2). Formation of 2D (SrBr)2PbI4was verified by small angle XRD peak at 6.4°, which corresponds to the layer distance of 13.78 Å. The best one of solar cells fabricated with Quasi-3D perovskite, (SrBr)2FA59Pb60I181(n= 60), showed a power conversion efficiency (PCE) of 18.46% and retained 95% of the initial PCE at 1000 h in the dry air. Further, the 2D (SrBr)2PbI4as the surface passivation layer on the 3D FAPbI3perovskite greatly reduced the defects at the perovskite/Spiro-OMeTAD interface, and the corresponding solar cells with FAPbI3/(SrBr)2PbI43D/2D structure achieved a PCE of 22.14% and over 90% retention of the original PCE at 1000 h. In short, this work provides an example of inorganic complex cations that can form 2D perovskites and achieve perovskite solar cells with high PCE and stabilization at the same time.

Strontium Chloride-Passivated Perovskite Thin Films for Efficient Solar Cells with Power Conversion Efficiency over 21% and Superior Stability.

Industrialization of perovskite solar cells is constrained by adverse stability in the air. Herein, we report effective strontium chloride (SrCl2) passivation upon HC(NH2)2-CH3NH3 (FA-MA)-based perovskite thin films for the suppression of nonradiative recombination. Moreover, the recombination dynamics, crystallinity, carrier transport, morphology, and the elemental stoichiometry of this film were systematically studied. By optimizing the concentration of SrCl2, the corresponding devices exhibited an increased open-circuit voltage (1.00 V vs 1.09 V), consistent with the enhanced photoluminescence lifetime. The champion passivated device showed an ascendant power conversion efficiency (PCE) about 21.11%, with over 90% retention of the primal PCE in dry air after 1000 h of aging with 20-30% humidity. A superior stability and an accelerated electron/hole-extraction ability were further observed by time-resolved photoluminescence spectroscopy.

[Pestle needle at Yaoyangguan bazhen and Heche-mingqiang section for 31 cases of bi syndrome at low back].

OBJECTIVE: To observe the clinical effect of pestle needle at Yaoyangguan bazhen and Heche-mingqiang section for bi syndrome (cold dampness type) at low back. METHODS: Thirty-one patients of bi syndrome (cold dampness type) at low back were treated with pestle needle at Yaoyangguan bazhen and Heche-mingqiang section. After the treatment of pestle needle, traditional acupuncture was used at Shenshu (BL 23), Dachangshu (BL 25), Huantiao (GB 30), Weizhong (BL 40), Chengshan (BL 57), Kunlun (BL 60), etc., followed by TCM washing at low back and legs. Once daily, five days of treatment were taken as one course, and there was an interval of 2 days between two courses. Totally 4 courses were given. The visual analogue scale (VAS) before and after treatment was observed, and the clinical efficacy was evaluated. RESULTS: The total effective rate was 90.3% (28/31) in 31 patients; the VAS after treatment was (2.42 ± 0.78), which was significantly, different from (5.59 ± 1.17) before treatment (P < 0.01). CONCLUSION The pestle needle at Yaoyangguan bazhen and Heche-mingqiang section is effective, noninvasive, painless for low-back bi syndrome (cold dampness type), and it is convenient for operation and easily accepted.