Prediction of potential invasion of two weeds of the genus Avena in Asia under climate change based on Maxent.

Avena sterilis L. (A. sterilis) and Avena ludoviciana Dur. (A. ludoviciana) are extremely invasive weeds with strong competitive ability and multiple transmission routes. Both species can invade a variety of dryland crops, including wheat, corn, and beans. Asia, as the world's major food-producing continent, will experience significant losses to agricultural production if it is invaded by these weeds on a large scale. This study used the MaxEnt model and ArcGIS to map the distribution of suitable habitats of the two species in Asia under climate change conditions. The constructed model comprised four levels, with a total of 25 index-level indicator factors used to evaluate the invasion risk of the two species. The results showed that the distribution of suitable habitats for both Avena species was highly dependent on precipitation and temperature. Under climate warming conditions, although overall the total suitable area is predicted to decrease compared to the current period, there are still moderately or highly suitable areas. Asian countries need to provide early warning for areas with significant increases in moderate and highly suitable zones for these two species of weeds under the background of climate change. If there is already an invaded area or if the suitability of the original area is increased, this should be closely monitored, and control measures should be taken to prevent further spread and deterioration.

Clinical nurses' work procrastination and smartphone addiction: a potential profile study.

Background: In the medical field, effective time management by clinical nurses is crucial for enhancing the quality of patient care. However, in recent years, with increasing work pressure for clinical nurses, procrastination has become a prevalent issue. Many nurses use smartphones as a way to alleviate stress and manage emotions, but excessive smartphone use could exacerbate procrastination, thereby jeopardizing patient safety and healthcare quality. Therefore, understanding the current state of work procrastination among clinical nurses, its heterogeneity, and exploring the impact of smartphone addiction and demographic factors on different aspects of nurse procrastination hold significant importance for improving patient care quality. Objective: This study aims to explore the current state of work procrastination among clinical nurses and identify potential profile categories. It further analyzes the impact of mobile phone addiction and demographic factors on work procrastination among clinical nurses. Methods: Convenience sampling was employed to recruit participants from three tertiary hospitals in central China from October to November 2023. Surveys measuring nurses' work procrastination and smartphone addiction were distributed and collected through online platforms. A total of 1,536 nurses participated in this study. Mplus 8.3 statistical software was used for latent profile analysis of clinical nurses' work procrastination, and SPSS 26.0 software was utilized for chi-square tests, rank-sum tests, and multi-classification logistic regression analyses. Results: The median total score for clinical nurses' work procrastination was 21.00 (17.00, 28.00), and three subgroups were identified: low procrastination (66.93%), medium-low procrastination (20.66%), and medium-high procrastination (12.41%). Additionally, logistic regression analysis revealed that smartphone addiction and department atmosphere were common influencing factors for medium-low and medium-high work procrastination. Hospitals with stricter management and nurses holding the position of head nurse were more likely to belong to the low work procrastination group. Nurses with higher incomes or those holding intermediate titles were more prone to medium-low work procrastination, while those experiencing career advancement difficulties were more likely to exhibit medium-high work procrastination (p < 0.05). Conclusion: Clinical nurses' work procrastination is generally at a medium-to-low level, with three subgroups identified: low procrastination, medium-low procrastination, and medium-high procrastination. Additionally, clinical nurses in surgical departments or those with intermediate titles exhibit higher levels of procrastination. Factors such as smartphone addiction, higher monthly income, tense departmental atmosphere, and unsuccessful career advancement are more likely to lead to work procrastination. Conversely, nurses in hospitals with strict management or those holding the position of head nurse exhibit lower levels of work procrastination. Therefore, nursing managers should pay close attention to the work procrastination behaviors of clinical nurses, actively monitor predictive factors among different groups, and provide psychological counseling and relevant training based on individual nurse circumstances. Additionally, it is also essential to focus on and improve departmental atmosphere and nurse smartphone addiction to enhance clinical nurses' work efficiency and reduce work procrastination.

Rolling up 2D WSe2 Nanosheets to 1D Anisotropic Nanoscrolls for Polarization-Sensitive Photodetectors.

The intrinsic low-symmetry crystal structures or external geometries of low-dimensional materials are crucial for polarization-sensitive photodetection. However, these inherently anisotropic materials are limited in variety, and their anisotropy is confined to specific crystal directions. Transforming 2D semiconductors, such as WSe2, from isotropic 2D nanosheets into anisotropic 1D nanoscrolls expands their application in polarization photodetection. Despite this considerable potential, research on polarization photodetection based on nanoscrolls remains scarce. Here, the uniform crystalline orientation of WSe2 nanoscrolls is achieved conveniently and efficiently by applying ethanol droplets to vapor deposition-grown bilayer WSe2 nanosheets. Angle-resolved polarized Raman spectroscopy of WSe2 nanoscrolls demonstrates vibrational anisotropy. Photodetectors based on these nanoscrolls show competitive overall performance with a broadband detection range from 405 to 808 nm, a competitive on/off ratio of ≈900, a high detectivity of 3.4 × 108 Jones, and a fast response speed of ≈30 ms. Additionally, WSe2 nanoscroll-based photodetectors exhibit strong polarization-sensitive detection with a maximum dichroic ratio of 1.5. More interestingly, due to high photosensitivity, the WSe2 nanoscroll detectors can easily record sequential puppy images. This work reveals the potential of WSe2 nanoscrolls as excellent polarization-sensitive photodetectors and provides new insights into the development of high-performance optoelectronic devices.

Misdiagnosis of hemangioma of left triangular ligament of the liver as gastric submucosal stromal tumor: Two case reports.

BACKGROUND: Extragastric lesions are typically not misdiagnosed as gastric submucosal tumor (SMT). However, we encountered two rare cases where extrinsic lesions were misdiagnosed as gastric SMTs. CASE SUMMARY: We describe two cases of gastric SMT-like protrusions initially misdiagnosed as gastric SMTs by the abdominal contrast-enhanced computed tomography (CT) and endoscopic ultrasound (EUS). Based on the CT and EUS findings, the patients underwent gastroscopy; however, no tumor was identified after incising the gastric wall. Subsequent surgical exploration revealed no gastric lesions in both patients, but a mass was found in the left triangular ligament of the liver. The patients underwent laparoscopic tumor resection, and the postoperative diagnosis was hepatic hemangiomas. CONCLUSION: During EUS procedures, scanning across different layers and at varying degrees of gastric cavity distension, coupled with meticulous image analysis, has the potential to mitigate the likelihood of such misdiagnoses.

The potential habitat of Angelica dahurica in China under climate change scenario predicted by Maxent model.

Since the 20th century, global climate has been recognized as the most important environmental factor affecting the distribution of plants. Angelica dahurica (A. dahurica) has been in great demand as a medicinal herb and flavoring, but the lack of seed sources has hindered its development. In this study, we utilized the Maxent model combined with Geographic Information System (GIS) to predict the potential habitat of A. dahurica in China based on its geographical distribution and 22 environmental factors. This prediction will serve as a valuable reference for the utilization and conservation of A. dahurica resources.The results indicated that: (1) the Maxent model exhibited high accuracy in predicting the potential habitat area of A. dahurica, with a mean value of the area under the ROC curve (AUC) at 0.879 and a TSS value above 0.6; (2) The five environmental variables with significant effects were bio6 (Min temperature of the coldest month), bio12 (Annual Precipitation), bio17 (Precipitation of Driest Quarter), elevation, and slope, contributing to a cumulative total of 89.6%. Suitable habitats for A. dahurica were identified in provinces such as Yunnan, Guizhou, Guangxi, Sichuan, Hunan, and others. The total area of suitable habitat was projected to increase, with expansion primarily in middle and high latitudes, while areas of decrease were concentrated in lower latitudes. Under future climate change scenarios, the centers of mass of suitable areas for A. dahurica were predicted to shift towards higher latitudes in the 2050s and 2090s, particularly towards the North China Plain and Northeast Plain. Overall, it holds great significance to utilize the Maxent model to predict the development and utilization of A. dahurica germplasm resources in the context of climate change.

Impact of health shocks on household consumption structure.

Introduction: In the aftermath of the pandemic, the impact of health shocks on household expenditure patterns has become a critical area of focus due to the heightened uncertainty surrounding consumers' expectations. Household medical expenditures have emerged as a key factor in the evolving consumption structure. Method: This research developed a practical framework to assess the influence of health shocks on family spending patterns, focusing on health shortfalls. Health emergencies were measured through randomized medical spending. Data was sourced from the 2010-2018 Wave 5 Statistical Report of the China Family Panel Studies (CFPS), which included a total of 25,809 participants. Results: The findings revealed that health shocks significantly increased the proportion of household spending on medical expenses. Concurrently, sub-expenditures such as food and education were reduced to varying degrees as households adjusted their consumption patterns to mitigate the impact of health shocks. The effects of these shocks were more pronounced in low-income households and those with health insurance. Discussions: The government should take steps to promote public health, reduce the burden of medical expenses resulting from health shocks, and unlock the consumption potential. Additionally, efforts should be made to boost economic growth and systematically upgrade household consumption patterns to effectively cushion the impact of health shocks.

Research perspective in the clinical management of Kawasaki disease.

This study examines research perspective in the clinical diagnosis, treatment, and prevention of cardiovascular complications in Kawasaki Disease (KD). Starting with an overview of the disease, it introduces KD's clinical manifestations, etiology, epidemiological features, and its impact on the cardiovascular system. Subsequently, the study discusses in detail the diagnostic methods, pathological mechanisms, and treatment strategies for KD, including foundational and emerging approaches such as high-dose intravenous immunoglobulin and aspirin therapy, biologic therapy, and corticosteroid pulse therapy. Additionally, it outlines strategies for preventing cardiovascular complications, including early risk assessment and long-term management. The study also explores the intersection of the COVID-19 pandemic with an increase in KD-like symptoms, emphasizing the need for further studies on the association between SARS-CoV-2 and KD. Lastly, it explores future research directions to enhance understanding of KD and improve patient outcomes and quality of life. This study provides valuable insights into the comprehensive treatment and management of KD and highlights avenues for future research.

High-Stability Composite Solid Polymer Electrolyte Composed of PAEPU/PP Nonwoven Fabric for Lithium-Ion Batteries.

Solid polymer electrolytes have attracted considerable attention, owing to their flexibility and safety. At present, poly(ethylene oxide) is the most widely studied polymer electrolyte matrix. It exhibits higher safety than the polyolefin diaphragm used in traditional lithium-ion batteries. However, it readily crystallizes at room temperature, resulting in low ionic conductivity, and the preparation process involves organic solvents. In this study, from the perspective of molecular design, solvent-free polyaspartate polyurea (PAEPU) and the cheap and easily available polypropylene (PP) nonwoven fabric were used as support materials for the PAEPU/PP composite solid polymer electrolyte (PAEPU/PP m -CPE). This CPE has good thermal stability, dimensional stability, flexibility, and mechanical properties. Among the different CPEs that were analyzed, PAEPU/PP10-CPE@20 had the highest ionic conductivity, which was reinforced with 10 g/m2 PP nonwoven fabric and the content of lithium salt was 20 wt %. Furthermore, PAEPU/PP10-CPE@20 exhibited the highest electrochemical stability with an electrochemical window value of 5.53 V. Moreover, the capacity retention rate of the Li//PAEPU/PP10-CPE@20//LiFePO4 half-cell was 96.82% after 150 cycles at 0.05 C and 60 °C, and the capacity recovery rate in the rate test reached 98.81%.

PD1+CD4+ T cells promote receptor editing and suppress autoreactivity of CD19+CD21low B cells within the lower respiratory airways in adenovirus pneumonia.

Human adenovirus (HAdV) pneumonia poses a major health burden for young children, however, factors that contribute to disease severity remain elusive. We analyzed immune cells from bronchoalveolar lavage (BAL) of children with HAdV pneumonia and found that CD19+CD21low B cells were significantly enriched in the BAL and were associated with increased autoantibody concentrations and disease severity. Myeloid cells, PD-1+CD4+ T helper cells and CD21low B cells formed tertiary lymphoid structures within the respiratory tracts. Myeloid cells promoted autoantibody production by expressing high amounts of B cell activating factor (BAFF). In contrast, PD-1+CD4+ T helper cells induced production of IgG1 and IgG3 antibodies but suppressed autoreactive IgGs by initiating B cell receptor editing. In summary, this study reveals cellular components involved in protective versus autoreactive immune pathways in the respiratory tract, and these findings provide potential therapeutic targets for severe HAdV lower respiratory tract infections.

Combination of light quality and melatonin regulates the quality in mustard sprouts.

Mustard sprouts is a new form of vegetable product that is gaining attention due to its high content of health-promoting compounds such as glucosinolates. This study investigated the effects of different light qualities (white, red, and blue) alone and in combination with 100 µmol L-1 melatonin on the growth and health-promoting substance content of mustard sprouts. The results showed that white light + melatonin treatment promoted the accumulation of glucosinolates in sprouts (compared with white light increased by 47.89%). The edible fresh weight of sprouts treated with red light + melatonin was the highest, followed by white light + melatonin treatment. In addition, the sprouts treated with blue light + melatonin contained more ascorbic acid, flavonoids, and total phenolics. Therefore, the combined treatment of light quality (especially white light) and melatonin can provide a new strategy to improve the quality of mustard sprouts.

Gut microbiota regulation of T lymphocyte subsets during systemic lupus erythematosus.

BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by disturbance of pro-inflammatory and anti-inflammatory lymphocytes. Growing evidence shown that gut microbiota participated in the occurrence and development of SLE by affecting the differentiation and function of intestinal immune cells. The purpose of this study was to investigate the changes of gut microbiota in SLE and judge its associations with peripheral T lymphocytes. METHODS: A total of 19 SLE patients and 16 HCs were enrolled in this study. Flow cytometry was used to detect the number of peripheral T lymphocyte subsets, and 16 s rRNA was used to detect the relative abundance of gut microbiota. Analyzed the correlation between gut microbiota with SLEDAI, ESR, ds-DNA and complement. SPSS26.0 software was used to analyze the experimental data. Mann-Whitney U test was applied to compare T lymphocyte subsets. Spearman analysis was used for calculating correlation. RESULTS: Compared with HCs, the proportions of Tregs (P = 0.001), Tfh cells (P = 0.018) and Naïve CD4 + T cells (P = 0.004) significantly decreased in SLE patients, and proportions of Th17 cells (P = 0.020) and γδT cells (P = 0.018) increased in SLE. The diversity of SLE patients were significantly decreased. Addition, there were 11 species of flora were discovered to be distinctly different in SLE group (P < 0.05). In the correlation analysis of SLE, Tregs were positively correlated with Ruminococcus2 (P = 0.042), Th17 cells were positively correlated with Megamonas (P = 0.009), γδT cells were positively correlated with Megamonas (P = 0.003) and Streptococcus (P = 0.004), Tfh cells were positively correlated with Bacteroides (P = 0.040), and Th1 cells were negatively correlated with Bifidobacterium (P = 0.005). As for clinical indicators, the level of Tregs was negatively correlated with ESR (P = 0.031), but not with C3 and C4, and the remaining cells were not significantly correlated with ESR, C3 and C4. CONCLUSION: Gut microbiota and T lymphocyte subsets of SLE changed and related to each other, which may break the immune balance and affect the occurrence and development of SLE. Therefore, it is necessary to pay attention to the changes of gut microbiota and provide new ideas for the treatment of SLE.

Gut mycobiome alterations in obesity in geographically different regions.

The gut fungi play important roles in human health and are involved in energy metabolism. This study aimed to examine gut mycobiome composition in obese subjects in two geographically different regions in China and to identify specific gut fungi associated with obesity. A total of 217 subjects from two regions with different urbanization levels [Hong Kong (HK): obese, n = 59; lean, n = 59; Kunming (KM): obese, n = 50; lean, n = 49. Mean body mass index (BMI) for obesity = 33.7] were recruited. We performed deep shotgun metagenomic sequencing on fecal samples to compare gut mycobiome composition and trophic functions in lean and obese subjects across these two regions. The gut mycobiome of obese subjects in both HK and KM were altered compared to those of lean subjects, characterized by a decrease in the relative abundance of Nakaseomyces, Schizosaccharomyces pombe, Candida dubliniensis and an increase in the abundance of Lanchanceathermotolerans, Saccharomyces paradox, Parastagonospora nodorum and Myceliophthorathermophila. Reduced fungal - bacterial and fungal - fungal correlations as well as increased negative fungal-bacterial correlations were observed in the gut of obese subjects. Furthermore, the anti-obesity effect of fungus S. pombe was further validated using a mouse model. Supplementing high-fat diet-induced obese mice with the fungus for 12 weeks led to a significant reduction in body weight gain (p < 0.001), and an improvement in lipid and glucose metabolism compared to mice without intervention. In conclusion, the gut mycobiome composition and functionalities of obese subjects were altered. These data shed light on the potential of utilizing fungus-based therapeutics for the treatment of obesity. S. pombe may serve as a potential fungal probiotic in the prevention of diet-induced obesity and future human trials are needed.

Inoculation with Funneliformis mosseae promotes selenium accumulation and transformation in pepper (Capsicum annuum L.).

Selenium (Se) is one of the fifteen essential nutrients required by the human body. Mycorrhizal microorganisms play a crucial role in enhancing selenium availability in plants. However, limited research exists on the impact of arbuscular mycorrhizal fungi (AMF) on selenium accumulation and transport in pepper plants. This study employed a pot experiment to investigate the changes in pepper plant growth, selenium accumulation, and transformation following inoculation with AMF and varying concentrations of exogenous selenium. The results indicate that exogenous selenium application in pepper has dual effects. At low concentrations (≤8 mg L⁻1), it promotes growth and nutrient accumulation, whereas high concentrations (>16 mg L⁻1) inhibit these processes. AMF inoculation positively influences selenium accumulation and transport in peppers, significantly increasing yield per plant by 17.89%, vitamin C content by 67.36%, flavonoid content by 43.26%, capsaicin content by 14.82%, DPPH radical scavenging rate by 18.18%, and ABTS radical scavenging rate by 27.81%. Additionally, it significantly reduces selenocysteine methyltransferase (SMT) enzyme activity, while minimally affecting ATP sulfurylase (ATPS) and adenosyl sulfate reductase (APR) enzyme activities. The combined treatment of AMF and 8 mg L⁻1 exogenous selenium has been proven to be the most effective for selenium enrichment in peppers, offering new insights into utilizing exogenous selenium and AMF inoculation to enhance selenium content in peppers.

ICAM-1 may promote the loss of dopaminergic neurons by regulating inflammation in MPTP-induced Parkinson's disease mouse models.

Parkinson's disease (PD) is a chronic neurodegenerative disease with unclear pathogenesis that involves neuroinflammation and intestinal microbial dysbiosis. Intercellular adhesion molecule-1 (ICAM-1), an inflammatory marker, participates in neuroinflammation during dopaminergic neuronal damage. However, the explicit mechanisms of action of ICAM-1 in PD have not been elucidated. We established a subacute PD mouse model by the intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and observed motor symptoms and gastrointestinal dysfunction in mice. Immunofluorescence was used to examine the survival of dopaminergic neurons, expression of microglial and astrocyte markers, and intestinal tight junction-associated proteins. Then, we use 16 S rRNA sequencing to identify alterations in the microbiota. Our findings revealed that ICAM-1-specific antibody (Ab) treatment relieved behavioural defects, gastrointestinal dysfunction, and dopaminergic neuronal death in MPTP-induced PD mice. Further mechanistic investigations indicated that ICAM-1Ab might suppress neuroinflammation by inhibiting the activation of astrocytes and microglia in the substantia nigra and relieving colon barrier impairment and intestinal inflammation. Furthermore, 16 S rRNA sequencing revealed that the relative abundances of bacterial Firmicutes, Clostridia, and Lachnospiraceae were elevated in the PD mice. However, ICAM-1Ab treatment ameliorated the MPTP-induced disorders in the intestinal microbiota. Collectively, we concluded that the suppressing ICAM-1 might lead to the a significant decrease of inflammation and restore the gut microbial community, thus ameliorating the damage of DA neurons.

Chromosome-scale genome assembly of oil-tea tree Camellia crapnelliana.

Camellia crapnelliana Tutch., belonging to the Theaceae family, is an excellent landscape tree species with high ornamental values. It is particularly an important woody oil-bearing plant species with high ecological, economic, and medicinal values. Here, we first report the chromosome-scale reference genome of C. crapnelliana with integrated technologies of SMRT, Hi-C and Illumina sequencing platforms. The genome assembly had a total length of ~2.94 Gb with contig N50 of ~67.5 Mb, and ~96.34% of contigs were assigned to 15 chromosomes. In total, we predicted 37,390 protein-coding genes, ~99.00% of which could be functionally annotated. The chromosome-scale genome of C. crapnelliana will become valuable resources for understanding the genetic basis of the fatty acid biosynthesis, and greatly facilitate the exploration and conservation of C. crapnelliana.

BoaBZR1.1 mediates brassinosteroid-induced carotenoid biosynthesis in Chinese kale.

Brassinazole resistant 1 (BZR1), a brassinosteroid (BR) signaling component, plays a pivotal role in regulating numerous specific developmental processes. Our study demonstrated that exogenous treatment with 2,4-epibrassinolide (EBR) significantly enhanced the accumulation of carotenoids and chlorophylls in Chinese kale (Brassica oleracea var. alboglabra). The underlying mechanism was deciphered through yeast one-hybrid (Y1H) and dual-luciferase (LUC) assays, whereby BoaBZR1.1 directly interacts with the promoters of BoaCRTISO and BoaPSY2, activating their expression. This effect was further validated through overexpression of BoaBZR1.1 in Chinese kale calli and plants, both of which exhibited increased carotenoid accumulation. Additionally, qPCR analysis unveiled upregulation of carotenoid and chlorophyll biosynthetic genes in the T1 generation of BoaBZR1.1-overexpressing plants. These findings underscored the significance of BoaBZR1.1-mediated BR signaling in regulating carotenoid accumulation in Chinese kale and suggested the potential for enhancing the nutritional quality of Chinese kale through genetic engineering of BoaBZR1.1.

Self-Rolled-Up WSe2 One-Dimensional/Two-Dimensional Homojunctions: Enabling High-Performance Self-Powered Polarization-Sensitive Photodetectors.

Mixed-dimensional heterostructures integrate materials of diverse dimensions with unique electronic functionalities, providing a new platform for research in electron transport and optoelectronic detection. Here, we report a novel covalently bonded one-dimensional/two-dimensional (1D/2D) homojunction structure with robust junction contacts, which exhibits wide-spectrum (from the visible to near-infrared regions), self-driven photodetection, and polarization-sensitive photodetection capabilities. Benefiting from the ultralow dark current at zero bias voltage, the on/off ratio and detectivity of the device reach 1.5 × 103 and 3.24 × 109 Jones, respectively. Furthermore, the pronounced anisotropy of the WSe2 1D/2D homojunction is attributed to its low symmetry, enabling polarization-sensitive detection. In the absence of any external bias voltage, the device exhibits strong linear dichroism for wavelengths of 638 and 808 nm, with anisotropy ratios of 2.06 and 1.96, respectively. These results indicate that such mixed-dimensional structures can serve as attractive building blocks for novel optoelectronic detectors.

Longitudinal dynamic single-cell mass cytometry analysis of peripheral blood mononuclear cells in COVID-19 patients within 6 months after viral RNA clearance.

This study investigates the longitudinal dynamic changes in immune cells in COVID-19 patients over an extended period after recovery, as well as the interplay between immune cells and antibodies. Leveraging single-cell mass spectrometry, we selected six COVID-19 patients and four healthy controls, dissecting the evolving landscape within six months post-viral RNA clearance, alongside the levels of anti-spike protein antibodies. The T cell immunophenotype ascertained via single-cell mass spectrometry underwent validation through flow cytometry in 37 samples. Our findings illuminate that CD8 + T cells, gamma-delta (gd) T cells, and NK cells witnessed an increase, in contrast to the reduction observed in monocytes, B cells, and double-negative T (DNT) cells over time. The proportion of monocytes remained significantly elevated in COVID-19 patients compared to controls even after six-month. Subpopulation-wise, an upsurge manifested within various T effector memory subsets, CD45RA + T effector memory, gdT, and NK cells, whereas declines marked the populations of DNT, naive and memory B cells, and classical as well as non-classical monocytes. Noteworthy associations surfaced between DNT, gdT, CD4 + T, NK cells, and the anti-S antibody titer. This study reveals the changes in peripheral blood mononuclear cells of COVID-19 patients within 6 months after viral RNA clearance and sheds light on the interactions between immune cells and antibodies. The findings from this research contribute to a better understanding of immune transformations during the recovery from COVID-19 and offer guidance for protective measures against reinfection in the context of viral variants.

[Mechanism of acupotomy on chondrocyte ferroptosis in rabbits with knee osteoarthritis based on HSPA5/GPX4 signaling pathway].

OBJECTIVE: To observe the effect of acupotomy on heat shock protein A family member 5 (HSPA5)/glutathione peroxidase 4 (GPX4) signaling pathway in the chondrocytes of the rabbits with knee osteoarthritis (KOA) and explore the mechanism of acupotomy on chondrocyte ferroptosis in KOA. METHODS: Twenty-seven New Zealand rabbits were randomly divided into a normal group, a model group and an acupotomy group, with 9 rabbits in each group. The left hind limb was fixed by the modified Videman method for 6 weeks to establish KOA model. After modeling, acupotomy was given in the acupotomy group, once a week and for consecutive 3 weeks. Using Lequesne MG score, the local symptoms, physical signs and functions of knee joint were evaluated. With HE staining and saffrane-solid green staining adopted, the morphology of chondrocytes and cartilage tissue was observed. Under transmission electron microscope, the mitochondrial structure of chondrocytes was observed. The iron content of cartilage tissue was detected by iron ion kit. The mitochondrial membrane potential (Δψm) and the reactive oxygen species (ROS) level in cartilage tissue were determined by flow cytometry, and the mitochondrial damage rate was calculated. The mRNA expression of HSPA5, GPX4, type Ⅱ collagen α1 chain (COL2A1), matrix metalloproteinases (MMP) 3 and MMP13 was detected by the real-time quantitative PCR; and the protein expression of HSPA5, GPX4, type Ⅱ collagen (COL-Ⅱ), MMP3 and MMP13 was detected by Western blot. The mean flourscence intensity of HSPA5 and GPX4 in cartilage tissue was determined by immunofluorescence. RESULTS: Before intervention, compared with the normal group, the Lequesne MG scores were increased in the model group and the acupotomy group (P<0.01). After intervention, the Lequesne MG score in the acupotomy group was decreased when compared with that in the model group. In comparison with that in the normal group, the number of chondrocytes was reduced and the cells were disarranged; the layers of cartilage structure were unclear, the tide lines disordered and blurred; the mitochondria were wrinkled and the mitochondrial crista decreased or even disappeared in the model group. Compared with the model group, the number of chondrocytes was increased, the layers of cartilage structure were clear, the tide lines recovered, the number of mitochondria elevated, with normal structure and more crista in the acupotomy group. The iron content of cartilage tissue was increased (P<0.01), the Δψm of chondrocytes was declined, the mitochondrial damage rate was increased (P<0.01), the average fluorescence intensity of ROS was increased (P<0.01); the mRNA and corresponding protein expression of HSPA5, GPX4 and COL2A1 was decreased (P<0.01), the mRNA and protein expression of MMP3 and MMP13 was increased (P<0.01) and the average fluorescence intensity of HSPA5, GPX4 was decreased (P<0.01) in the model group when compared with those in the normal group. Compared with the model group, the iron content in cartilage tissue was reduced (P<0.01), the Δψm of chondrocytes was increased, the mitochondrial damage rate was decreased (P<0.01), and the average fluorescence intensity of ROS was decreased (P<0.01); the mRNA and corresponding protein expression of HSPA5, GPX4 and COL2A1 was higher (P<0.01), and the mRNA and protein expression of MMP3 and MMP13 was lower, and the average fluorescence intensity of HSPA5, GPX4 was increased (P<0.01) in the acupotomy group. CONCLUSION: Acupotomy can alleviate cartilage injury of KOA rabbits, and its mechanism may be related to the regulation of HSPA5/GPX4 signaling pathway to maintain iron homeostasis in articular cartilage, thus inhibiting chondrocyte ferroptosis and relieving extracellular matrix degradation.

A GMP-compliant manufacturing method for Wharton's jelly-derived mesenchymal stromal cells.

BACKGROUND: Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) hold great therapeutic potential in regenerative medicine. Therefore, it is crucial to establish a Good Manufacturing Practice (GMP)-compliant methodology for the isolation and culture of WJ-MSCs. Through comprehensive research, encompassing laboratory-scale experiments to pilot-scale studies, we aimed to develop standardized protocols ensuring the high yield and quality of WJ-MSCs manufacturing. METHODS: Firstly, optimization of parameters for the enzymatic digestion method used to isolate WJ-MSCs was conducted. These parameters included enzyme concentrations, digestion times, seeding densities, and culture media. Additionally, a comparative analysis between the explant method and the enzymatic digestion method was performed. Subsequently, the consecutive passaging of WJ-MSCs, specifically up to passage 9, was evaluated using the optimized method. Finally, manufacturing processes were developed and scaled up, starting from laboratory-scale flask-based production and progressing to pilot-scale cell factory-based production. Furthermore, a stability study was carried out to assess the storage and use of drug products (DPs). RESULTS: The optimal parameters for the enzymatic digestion method were a concentration of 0.4 PZ U/mL Collagenase NB6 and a digestion time of 3 h, resulting in a higher yield of P0 WJ-MSCs. In addition, a positive correlation between the weight of umbilical cord tissue and the quantities of P0 WJ-MSCs has been observed. Evaluation of different concentrations of human platelet lysate revealed that 2% and 5% concentrations resulted in similar levels of cell expansion. Comparative analysis revealed that the enzymatic digestion method exhibited faster outgrowth of WJ-MSCs compared to the explant method during the initial passage. Passages 2 to 5 exhibited higher viability and proliferation ability throughout consecutive passaging. Moreover, scalable manufacturing processes from the laboratory scale to the pilot scale were successfully developed, ensuring the production of high-quality WJ-MSCs. Multiple freeze-thaw cycles of the DPs led to reduced cell viability and viable cell concentration. Subsequent thawing and dilution of the DPs resulted in a significant decrease in both metrics, especially when stored at 20-27 °C. CONCLUSION: This study offers valuable insights into optimizing the isolation and culture of WJ-MSCs. Our scalable manufacturing processes facilitate the large-scale production of high-quality WJ-MSCs. These findings contribute to the advancement of WJ-MSCs-based therapies in regenerative medicine.