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Soil seed bank (SSB) play an essential role in the restoration and regeneration of plant communities. Grazing is a common disturbance factor that exerts an important effect on the diversity and composition of the SSB community in forests. Here, we evaluated the structure and composition of SSB along a grazing disturbance gradient of two types of vegetation (broad-leaved forests and mixed coniferous broad-leaved forests) in southwest China. A comparison of the similarity of SSBs with the above-ground vegetation strata (tree, shrub and herb layers) was also analyzed. The experiment revealed the germination of 25 species that belonged to 14 families and 24 genera. The density of SSBs did not show significant differences along the grazing disturbance. The alpha diversity of SSBs was the highest at moderate grazing intensity in broad-leaved forests, and not significantly different along disturbance intensities in mixed coniferous broad-leaved forests. No significant changes were observed in the species composition of SSB along the grazing gradients. Nonmetric multidimensional scaling analyses revealed low similarities between SSBs and above-ground vegetation, with the lowest similarity occurring under medium grazing disturbance intensity. Overall, our research offers significant insight into the response of SSBs to grazing in different vegetation types. It serves as a guide for designing grazing management strategies tailored to natural forests.

Background: The chondrogenic differentiation of mesenchymal stem cells (MSCs) to enhance cartilage repair and regeneration is a promising strategy to alleviate osteoarthritis (OA) progression. Method: The potency of JD-312 in inducing chondrogenic differentiation of MSCs was assessed and verified. The efficacy of JD-312-treated MSCs was evaluated using a Sprague–Dawley rat DMM model. Additionally, the capacity of JD-312 to successfully recruit bone marrow-derived mesenchymal stem cells (BMSCs) for the treatment of OA in vitro was confirmed via intra-articular injection. The repair status of the articular cartilage was analyzed in vivo through histological examination. Result: In this study, we identify JD-312 as a novel non-toxic small molecule that can promote chondrogenic differentiation in human umbilical cord-derived MSCs (hUCMSCs) and human bone marrow MSCS (hBMSCs) in vitro. We also show that transient differentiation of MSCs with JD-312 prior to in vivo administration remarkably improves the regeneration of cartilage and promotes Col2a1 and Acan expression in rat models of DMM, in comparison to kartogenin (KGN) pre-treatment or MSCs alone. Furthermore, direct intra-articular injection of JD-312 in murine model of OA showed reduced loss of articular cartilage and improved pain parameters. Lastly, we identified that the effects of JD-312 are at least in part mediated via upregulation of genes associated with the focal adhesion, PI3K-Akt signaling and the ECM-receptor interaction pathways, and specifically cartilage oligomeric matrix protein (COMP) may play a vital role. Conclusion: Our study demonstrated that JD-312 showed encouraging repair effects for OA in vivo. The translational potential of this article: Together, our findings demonstrate that JD-312 is a promising new therapeutic molecule for cartilage regeneration with clinical potential.

Gut microbes are pivotal reference indicators for assessing the health status of animals. Before introducing artificially bred species into the wild, examining their gut microbe composition is crucial to help mitigate potential threats posed to wild populations. However, gut microbiological trait similarities between wild and artificially bred green turtles remain unexplored. Therefore, this study compared the gut microbiological characteristics of wild and artificially bred green turtles (Chelonia mydas) through high-throughput Illumina sequencing technology. The α-diversity of intestinal bacteria in wild green turtles, as determined by Shannon and Chao indices, significantly surpasses that of artificial breeding green turtles (p

Abstract Pluripotent stem cell‐derived small extracellular vesicles (PSC‐sEVs) have demonstrated great clinical translational potential in multiple aging‐related degenerative diseases. Characterizing the PSC‐sEVs is crucial for their clinical applications. However, the specific marker pattern of PSC‐sEVs remains unknown. Here, the sEVs derived from two typical types of PSCs including induced pluripotent stem cells (iPSC‐sEVs) and embryonic stem cells (ESC‐sEVs) were analysed using proteomic analysis by liquid chromatography with tandem mass spectrometry (LC‐MS/MS), and surface marker phenotyping analysis by nanoparticle flow cytometry (NanoFCM). A group of pluripotency‐related proteins were found to be enriched in PSC‐sEVs by LC‐MS/MS and then validated by Western Blot analysis. To investigate whether these proteins were specifically expressed in PSC‐sEVs, sEVs derived from seven types of non‐PSCs (non‐PSC‐sEVs) were adopted for analysis. The results showed that PODXL, OCT4, Dnmt3a, and LIN28A were specifically enriched in PSC‐sEVs but not in non‐PSC‐sEVs. Then, commonly used surface antigens for PSC identification (SSEA4, Tra‐1‐60 and Tra‐1‐81) and PODXL were gauged at single‐particle resolution by NanoFCM for surface marker identification. The results showed that the positive rates of PODXL (>50%) and SSEA4 (>70%) in PSC‐sEVs were much higher than those in non‐PSC‐sEVs (

ABSTRACT Sputum collection and testing for tuberculosis (TB) have been problematic due to the potential for aerosolization, difficulty in generating a quality sample, and complex DNA extraction methods. Tongue swabs are inexpensive, minimally invasive, and a promising alternative to sputum collection. We investigated the diagnostic accuracy of tongue swabs for TB diagnosis using the Truenat MTB Plus assay from Molbio Diagnostics with a direct to PCR processing method. Four tongue swabs were collected per participant using two nylon-flocked swabs and two spun polyester swabs. Following tongue swab sample collection, participants also provided two sputum samples, which were tested by Cepheid Xpert MTB/RIF Ultra or culture. Of the 81 participants enrolled, 24 (30%) were positive for TB disease by sputum Ultra. Using the Truenat MTB Plus test, tongue swabs had 54% (52/96) sensitivity and 99% (218/220) specificity compared to sputum Ultra. Crude lysate was also tested using an in-house qPCR assay, which allowed for increased sample input. Using this method, tongue swabs had 70% (67/96) sensitivity and 94% (216/224) specificity. Mycobacterium tuberculosis (MTB) sample quantification using digital PCR yielded between 20 copies (minimum) and 34,000 copies (maximum) of MTB per swab. In addition, serially collected tongue swabs resulted in similar levels of detected MTB, and spun polyester swabs performed equivalently to nylon-flocked swabs. Overall, this study demonstrates that tongue swab samples are compatible with the Truenat MTB testing platform and that a direct to PCR method is a viable diagnostic solution. IMPORTANCE Tuberculosis (TB) remains one of the world’s leading infectious disease killers, despite available treatments. Although highly sensitive molecular diagnostics are available, expensive equipment and poor infrastructure have hindered their implementation in low-resource settings. Furthermore, the collection of sputum poses challenges as it is difficult for patients to produce and creates dangerous aerosols. This manuscript explores tongue swabs as a promising alternative to sputum collection. While previous studies have explored the sensitivity of tongue swabs as compared to sputum, existing literature has not addressed the need to standardize and simplify laboratory processing for easy implementation in high TB burden areas. This manuscript provides the first evidence that detection of TB from a tongue swab is possible without the use of DNA extraction or purification steps. The data provided in this manuscript will improve the collection and testing of tongue swabs for the diagnosis of TB disease.

Objective: To assess the quality of systematic reviews assessing the effects of traditional Chinese exercises on controlling blood indices, anthropometric indices, psychological indices, and quality of life in patients with diabetes. Methods: Systematic studies/meta-analyses of intervention with traditional Chinese exercises on diabetes mellitus were searched in the PubMed, Web of Science, the China National Knowledge Infrastructure Databases (CNKI), Wan Fang Database, and Chinese Scientific Journal Database (VIP). Two researchers independently screened the studies and extracted the data. The methodology and quality of evidence of the included studies were assessed using A Measurement Tool to Assess Systematic Reviews 2 (AMSTAR-2) and Grading of Recommendations Assessment, Development and Evaluation (GRADE) criteria, respectively. Results: A total of 32 systematic reviews/meta-analyses were included in the present study. Of them, the methodological quality of 4 studies was graded as low, while that of the other 28 was graded as extremely low. The most common quality flaws in key items included a lack of preliminary proposals for systematic reviews, failure to explain the reasons for the inclusion criteria, failure to provide a list of excluded studies and reasons for exclusion, failure to report potential conflicts of interest, and inadequate assessment of publication bias. The quality of evidence for most of the 18 outcomes was subsequently graded as medium or low. Overall, the results of these studies indicated that Tai Chi, health qigong, and other traditional Chinese exercises lowered fasting blood glucose (FBG), 2-h postprandial blood glucose (2hPBG), hemoglobin A1c (HbA1c), and body mass index, and relieved anxiety and depression in patients with diabetes. Conclusion: Our findings indicate that the methodological quality of systematic reviews related to traditional Chinese exercises in the diabetic population is generally low, and the quality of evidence is also relatively poor. Therefore, we suggest that the quality of systematic reviews and meta-analyses on traditional Chinese exercises for controlling diabetes mellitus needs to be improved. In the future, researchers should conduct higher-quality clinical studies with reference to the AMSTAR2 checklist and GRADE system.

Osteoarthritis (OA) is one of the most common chronic diseases in the world. However, current treatment modalities mainly relieve pain and inhibit cartilage degradation, but do not promote cartilage regeneration. In this study, we show that G protein-coupled receptor class C group 5 member B (GPRC5B), an orphan G-protein-couple receptor, not only inhibits cartilage degradation, but also increases cartilage regeneration and thereby is protective against OA. We observed that Gprc5b deficient chondrocytes had an upregulation of cartilage catabolic gene expression, along with downregulation of anabolic genes in vitro. Furthermore, mice deficient in Gprc5b displayed a more severe OA phenotype in the destabilization of the medial meniscus (DMM) induced OA mouse model, with upregulation of cartilage catabolic factors and downregulation of anabolic factors, consistent with our in vitro findings. Overexpression of Gprc5b by lentiviral vectors alleviated the cartilage degeneration in DMM-induced OA mouse model by inhibiting cartilage degradation and promoting regeneration. We also assessed the molecular mechanisms downstream of Gprc5b that may mediate these observed effects and identify the role of protein kinase B (AKT)-mammalian target of rapamycin (mTOR)-autophagy signaling pathway. Thus, we demonstrate an integral role of GPRC5B in OA pathogenesis, and activation of GPRC5B has the potential in preventing the progression of OA.

Freshwater acidification (FA) has become a global environmental problem, posing a potential threat to freshwater ecosystems. The gut microbiota plays a crucial role in the host’s response and adaptation to new environments. In this study, we investigated the changes in microbial communities in Red-eared slider (Trachemys scripta elegans) under acidic conditions to reveal the ecological impacts of acidification on freshwater turtles. The results showed that there were significant differences in β-diversity (p = 0.03), while there were no significant differences in the α-diversity of gut microbiota in T. s. elegans between the different levels of acidification (pH of 5.5, 6.5, 7.5). Both the Gut Microbiome Health Index (GMHI) and the Microbial Dysbiosis Index (MDI) exhibited significant differences when comparing environments with a pH of 5.5 to those with a pH of 6.5 (p < 0.01). A comparative analysis between pH levels of 5.5 and 6.5 also revealed substantial differences (p < 0.01). Likewise, a comparative analysis between pH levels of 6.5 and 7.5 also revealed substantial differences (p < 0.01). At the phylum level, Firmicutes, Fusobacteria, and Bacteroidota formed a major part of the gut microbial community, Fusobacteria showed significant differences in different acidity environments (p = 0.03). At the genus level, Cetobacterium, Turicibacter, unclassified Eubacteriaceae, and Anaerorhabdus_furcosa_group showed significant differences in different acidity environments. The pH reduced interactivity in the gut microbiota of T. s. elegans. In addition, LEfSe analysis and functional prediction revealed that the potentially_pathogenic and stress_tolerant functional characteristics also showed significant differences in different acidity environments. The findings underscore the pivotal role of the gut microbiota in T. s. elegans in response to freshwater acidification and provide a foundation for further exploration into the impacts of acidification on freshwater ecosystems.

Abstract Deubiquitylating enzymes (DUBs) maintain relative homeostasis of the cellular ubiquitome by removing the post‐translational modification ubiquitin moiety from substrates. Numerous DUBs have been demonstrated specificity for cleaving a certain type of ubiquitin linkage or positions within ubiquitin chains. Moreover, several DUBs perform functions through specific protein–protein interactions in a catalytically independent manner, which further expands the versatility and complexity of DUBs’ functions. Dysregulation of DUBs disrupts the dynamic equilibrium of ubiquitome and causes various diseases, especially cancer and immune disorders. This review summarizes the Janus‐faced roles of DUBs in cancer including proteasomal degradation, DNA repair, apoptosis, and tumor metastasis, as well as in immunity involving innate immune receptor signaling and inflammatory and autoimmune disorders. The prospects and challenges for the clinical development of DUB inhibitors are further discussed. The review provides a comprehensive understanding of the multi‐faced roles of DUBs in cancer and immunity.

IntroductionThe adverse effects of high glucose on embryos can be traced to the preimplantation stage. This study aimed to observe the effect of high glucose on early-stage embryos. Methods and resultsSeven-week-old ICR female mice were superovulated and mated, and the zygotes were collected. The zygotes were randomly cultured in 5 different glucose concentrations (control, 20mM, 40mM, 60mM and 80mM glucose). The cleavage rate, blastocyst rate and total cell number of blastocyst were used to assess the embryo quality. 40 mM glucose was selected to model high glucose levels in this study. 40mM glucose arrested early embryonic development, and the blastocyst rate and total cell number of the blastocyst decreased significantly as glucose concentration was increased. The reduction in the total cell number of blastocysts in the high glucose group was attributed to decreased proliferation and increased cell apoptosis, which is associated with the diminished expression of GLUTs (GLUT1, GLUT2, GLUT3). Furthermore, the metabolic characterization of blastocyst culture was observed in the high-glucose environment. DiscussionThe balance of glycolysis and oxidative phosphorylation at the blastocyst stage was disrupted. And embryo development arrest due to high glucose is associated with changes in glycolysis and oxidative phosphorylation, as well as abnormalities in the TCA cycle and amino acid metabolism.

Lyophyllum decastes is a mushroom that is highly regarded for its culinary and medicinal properties. Its delectable taste and texture make it a popular choice for consumption. To gain a deeper understanding of the molecular mechanisms involved in the development of the fruiting body of L. decastes, we used RNA sequencing to conduct a comparative transcriptome analysis. The analysis encompassed various developmental stages, including the vegetative mycelium, primordial initiation, young fruiting body, medium-size fruiting body, and mature fruiting body stages. A range of 40.1 to 60.6 million clean reads were obtained, and de novo assembly generated 15,451 unigenes with an average length of 1,462.68 bp. Functional annotation of transcriptomes matched 76.84% of the unigenes to known proteins available in at least one database. The gene expression analysis revealed a significant number of differentially expressed genes (DEGs) between each stage. These genes were annotated and subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses. Highly differentially expressed unigenes were also identified, including those that encode extracellular enzymes, transcription factors, and signaling pathways. The accuracy of the RNA-Seq and DEG analyses was validated using quantitative PCR. Enzyme activity analysis experiments demonstrated that the extracellular enzymes exhibited significant differences across different developmental stages. This study provides valuable insights into the molecular mechanisms that underlie the development of the fruiting body in L. decastes.

Abstract Biomolecular condensates are cellular structures composed of membraneless assemblies comprising proteins or nucleic acids. The formation of these condensates requires components to change from a state of solubility separation from the surrounding environment by undergoing phase transition and condensation. Over the past decade, it has become widely appreciated that biomolecular condensates are ubiquitous in eukaryotic cells and play a vital role in physiological and pathological processes. These condensates may provide promising targets for the clinic research. Recently, a series of pathological and physiological processes have been found associated with the dysfunction of condensates, and a range of targets and methods have been demonstrated to modulate the formation of these condensates. A more extensive description of biomolecular condensates is urgently needed for the development of novel therapies. In this review, we summarized the current understanding of biomolecular condensates and the molecular mechanisms of their formation. Moreover, we reviewed the functions of condensates and therapeutic targets for diseases. We further highlighted the available regulatory targets and methods, discussed the significance and challenges of targeting these condensates. Reviewing the latest developments in biomolecular condensate research could be essential in translating our current knowledge on the use of condensates for clinical therapeutic strategies.

Abstract Gene dosage disorders (GDDs) constitute a major class of genetic risks for psychopathology, but there is considerable debate regarding the extent to which different GDDs induce different psychopathology profiles. The current research speaks to this debate by compiling and analyzing dimensional measures of several autism-related traits (ARTs) across seven diverse GDDs. The sample included 350 individuals with one of 7 GDDs, as well as reference idiopathic autism spectrum disorder (ASD; n = 74) and typically developing control (TD; n = 171) groups. The GDDs were: Down, Williams–Beuren, and Smith–Magenis (DS, WS, SMS) syndromes, and varying sex chromosome aneuploidies (“plusX”, “plusXX”, “plusY”, “plusXY”). The Social Responsiveness Scale (SRS-2) was used to measure ARTs at different levels of granularity—item, subscale, and total. General linear models were used to examine ART profiles in GDDs, and machine learning was used to predict genotype from SRS-2 subscales and items. These analyses were completed with and without covariation for cognitive impairment. Twelve of all possible 21 pairwise GDD group contrasts showed significantly different ART profiles (7/21 when co-varying for IQ, all Bonferroni-corrected). Prominent GDD–ART associations in post hoc analyses included relatively preserved social motivation in WS and relatively low levels of repetitive behaviors in plusX. Machine learning revealed that GDD group could be predicted with plausible accuracy (~60–80%) even after controlling for IQ. GDD effects on ARTs are influenced by GDD subtype and ART dimension. This observation has consequences for mechanistic, clinical, and translational aspects of psychiatric neurogenetics.

Abstract Objective To determine the benefits of aquatic physical therapy as a rehabilitation strategy for knee osteoarthritis patients. Methods Electronic databases systematically searched up to July 2021. Results 580 RCTs were selected. A total of thirteen studies comprising 883 participants were included in the study. For pain, meta-analyses showed that aquatic physical therapy is associated with a significant change in Western Ontario and McMaster University Osteoarthritis Index (WOMAC) pain (SMD = − 1.09, 95%CI − 1.97, − 0.21, p = 0.02) and visual analog scale (VAS) (SMD = − 0.55, 95%CI − 0.98, − 0.12, p = 0.01). In addition, for physical function, meta-analyses showed that aquatic physical therapy effectively improved WOMAC physical function (SMD = − 0.57, 95%CI − 1.14, − 0.01, p = 0.05). However, our findings showed no significant improvements in symptoms of joints, quality of life (QOL), flexibility, and body composition with knee osteoarthritis. For muscle strength, we found that aquatic physical therapy can only improve knee extension muscle strength (MD = 2.11, 95%CI 0.02, 4.20, p = 0.05). Additionally, for walking ability, we observed that aquatic physical therapy effectively reduced Timed-Up-and-Go Test (TUGT) in a large degree (MD = − 0.89, 95%CI − 1.25, − 0.53, p

Abstract Inflammatory diseases decrease the extracellular environmental pH. However, whether proton-activated G protein-coupled receptors (GPCRs) can regulate the development of osteoarthritis (OA) is largely unknown. In this study, we report that proton-activated GPR4 is essential for OA development. We found a marked increase in expression of the proton-activated GPR4 in human and mouse OA cartilage. Lentivirus-mediated overexpression of GPR4 in mouse joints accelerated the development of OA, including promotion of articular cartilage damage, synovial hyperplasia, and osteophyte formation, while Gpr4 knockout effectively attenuated the development of posttraumatic and aging-associated OA in mice. We also found that inhibition of GPR4 with the antagonist NE52-QQ57 ameliorated OA progression in mice, promoted extracellular matrix (ECM) production, and protected cartilage from degradation in human articular cartilage explants. Moreover, GPR4 overexpression upregulated matrix-degrading enzymes’ expression and inflammation factors under pro-inflammatory and slightly acidic conditions. Mechanistically, GPR4 suppressed chondrocyte differentiation and upregulated cartilage homeostasis through NF-κB/MAPK signaling activation by regulating CXCR7/CXCL12 expression. Together, our results take the lead to illustrate that proton-activated GPCR acts as a key regulator for OA pathogenesis in vivo, and support that GPR4 could be a promising therapeutic target for OA treatment.

Neuroimaging-based brain-age estimation via machine learning has emerged as an important new approach for studying brain aging. The difference between one's estimated brain age and chronological age, the brain age gap (BAG), has been proposed as an Alzheimer's Disease (AD) biomarker. However, most past studies on the BAG have been cross-sectional. Quantifying longitudinal changes in an individual's BAG temporal pattern would likely improve prediction of AD progression and clinical outcome based on neurophysiological changes. To fill this gap, our study conducted predictive modeling using a large neuroimaging dataset with up to 8 years of follow-up to examine the temporal patterns of the BAG's trajectory and how it varies by subject-level characteristics (sex, APOEɛ4 carriership) and disease status. Specifically, we explored the pattern and rate of change in BAG over time in individuals who remain stable with normal cognition or mild cognitive impairment (MCI), as well as individuals who progress to clinical AD. Combining multimodal imaging data in a support vector regression model to estimate brain age yielded improved performance over single modality. Multilevel modeling results showed the BAG followed a linear increasing trajectory with a significantly faster rate in individuals with MCI who progressed to AD compared to cognitively normal or MCI individuals who did not progress. The dynamic changes in the BAG during AD progression were further moderated by sex and APOEɛ4 carriership. Our findings demonstrate the BAG as a potential biomarker for understanding individual specific temporal patterns related to AD progression.

Abstract Grading individual knee osteoarthritis (OA) features is a fine-grained knee OA severity assessment. Existing methods ignore following problems: (1) more accurately located knee joints benefit subsequent grades prediction; (2) they do not consider knee joints’ symmetry and semantic information, which help to improve grades prediction performance. To this end, we propose a SE-ResNext50-32x4d-based Siamese network with adaptive gated feature fusion method to simultaneously assess eight tasks. In our method, two cascaded small convolution neural networks are designed to locate more accurate knee joints. Detected knee joints are further cropped and split into left and right patches via their symmetry, which are fed into SE-ResNext50-32x4d-based Siamese network with shared weights, extracting more detailed knee features. The adaptive gated feature fusion method is used to capture richer semantic information for better feature representation here. Meanwhile, knee OA/non-knee OA classification task is added, helping extract richer features. We specially introduce a new evaluation metric (top±1 accuracy) aiming to measure model performance with ambiguous data labels. Our model is evaluated on two public datasets: OAI and MOST datasets, achieving the state-of-the-art results comparing to competing approaches. It has the potential to be a tool to assist clinical decision making.

Soil inorganic carbon (SIC) is an essential component of the soil carbon pool and plays a vital role in the global carbon cycle and climate change. However, few studies have examined the effects of different agronomic practices on the SIC content. This paper aims to study the effects of different agronomic practices on the SIC content in the 0- to 40-cm soil layer of dryland fields; the innovations are intended to explore the relative importance and synergistic effects of different agronomic practices on the SIC content of the plough layer of dryland fields. We screened out 136 peer-reviewed articles worldwide from 1990–2022, with a total of 2612 valid data pairs, using meta-analysis to assess the effects of different agronomic practices on the SIC content of the plough layer of dryland fields. Compared to conventional tillage (CT), both no-tillage (NT) and plastic film mulching (PM) were able to increase the SIC content in the 0- to 40-cm soil layer of dryland fields. NT increased the SIC content by 15.07 ± 3.48%, while PM gradually increased SIC accumulation as the soil layer deepened, with the greatest increase in SIC content in the 30- to 40-cm soil layer at 11.61 ± 5.89%. When organic manure application (M) and straw return mulching (SM) were applied, the SIC content in the 0- to 40-cm soil layer of dryland fields showed a non-significant reduction trend, with the largest changes in SIC content in the 0- to 10-cm soil layer, at 5.23 ± 2% and 4.69 ± 3.53% reductions, respectively. No-tillage straw return (NTS) significantly increased the SIC content in the 0- to 40-cm soil layer of dryland fields by 77.34 ± 5.6%, which was significantly higher than the independent effects of NT and SM, showing a more substantial synergistic effect. Different agronomic practices have different effects on the SIC content of dryland fields in the 0- to 40-cm soil layer, with NTS > NT > PM > SM > M. Among practices, NTS, NT and PM can increase the SIC content of the plough layer of dryland fields; in particular, NTS can increase the SIC content of 0- to 40-cm in dryland fields to the greatest extent through a synergistic effect. At the same time, SM and M showed an insignificant reduction effect.

Therapeutics targeting osteoclasts are commonly used treatments for bone metastasis; however, whether and how osteoclasts regulate premetastatic niche and bone tropism are largely unknown. In this study, we report that osteoclast precursors (OPs) can function as a premetastatic niche component that facilitates breast cancer (BCa) bone metastasis at early stages. At the molecular level, unbiased GPCR ligand/agonist screening in BCa cells suggested that R-spondin 2 (RSPO2) and RANKL, through interaction with their receptor LGR4, promoted osteoclastic premetastatic niche formation and enhanced BCa bone metastasis. This was achieved by RSPO2/RANKL-LGR4 signal modulating the WNT inhibitor DKK1 through Gαq and β-catenin signaling. DKK1 directly facilitated OP recruitment through suppression of its receptor LDL receptor–related protein 5 (LRP5) but not LRP6, upregulating Rnasek expression via inhibition of canonical WNT signaling. In clinical samples, RSPO2, LGR4, and DKK1 expression showed a positive correlation with BCa bone metastasis. Furthermore, soluble LGR4 extracellular domain (ECD) protein, acting as a decoy receptor for RSPO2 and RANKL, significantly alleviated bone metastasis and osteolytic lesions in a mouse bone metastasis model. These findings provide unique insights into the functional role of OPs as key components of the premetastatic niche for BCa bone metastasis and identify RSPO2/RANKL-LGR4 signaling as a promising target for inhibiting BCa bone metastasis.

Objective: To investigate the effects of Ganjiang granule (GG) on cecal microflora and serum biochemical components in rats with constipation-predominant irritable bowel syndrome (IBS–C). Methods: Twenty-four Sprague–Dawley rats were randomly divided into four groups: control, model, GG, and probiotics. Rats in the model, GG, and probiotics groups received 3 °C tap water intragastrically; rats in the GG group were treated with GG; rats in the probiotics group were treated with probiotics. For all rats, enzyme-linked immunosorbent assay and colorimetry were used to assess serum biochemical components related to gastrointestinal function; 16S rRNA high-throughput sequencing was used to analyze the cecal microflora. Results: The serum level of 5-hydroxytryptamine (HT) was higher in the model group than in the control group (Z = −2.082, P = .037). The model group exhibited changes in cecal microflora: the relative abundances of Lactobacillus decreased (Z = −2.882, P = .004) and Dorea increased (t = −3.030, P = .023), compared with the control group. The GG and probiotics groups exhibited normal serum levels of 5-HT. The GG and probiotics groups exhibited improved serum levels of gastrin; the probiotics group exhibited an improved serum level of vasoactive intestinal peptide. Compared with the model group, The GG group exhibited greater relative abundance of Ruminococcus (Z = −2.402, P = .016); the probiotics group exhibited greater relative abundance of SMB53 (Z = −2.823, P = .005) and lower relative abundances of Desulfovibrio (Z = −2.823, P = .005) and Facklamia (Z = −2.608, P = .009). Conclusion: The effects of GG on IBS-C may be related to regulation of the serum level of 5-HT, as well as elevated relative abundance of beneficial bacteria in cecal microflora.

Abstract Background Premature ovarian failure (POF) has a great impact on reproductive endocrine function in females, and it is an important cause of infertility. Previous studies have demonstrated that small extracellular vesicles (sEVs) derived from stem cells play an important role in tissue regeneration. This study aimed to investigate the therapeutic effect of sEVs derived from embryonic stem cells (ESCs-sEVs) on damaged ovaries and explore the underlying molecular mechanisms. Methods Mice POF models were established by injecting mice with cyclophosphamide and busulfan. Then, ESCs-sEVs were intravenously transplanted into POF mice. The plasma of mice was harvested at 1 and 2 weeks after treatment to analyze the levels of anti-Mullerian hormone (AMH), estradiol (E2), and follicle stimulating hormone (FSH) by ELISA. The morphology of ovaries and follicles was observed by H&E staining, and apoptosis of granulosa cells was detected by TUNEL. In vitro, EdU and CCK-8 tests were used to evaluate the proliferation of cultured granulosa cells stimulated by ESCs-sEVs. Western blotting was used to determine the expression of PI3K/AKT and apoptotic-related proteins. Results After transplantation of ESCs-sEVs, the levels of serum sex hormones recovered to normal levels. In addition, the number of follicles was significantly increased, and the number of apoptotic cells was decreased. The results in vitro revealed that ESCs-sEVs could significantly improve the proliferation rate of granulosa cells and increase the expression of phosphorylated PI3K and AKT. Meanwhile, the positive effect on proliferation and the negative effect on apoptosis observed in granulosa cells were obviously decreased when the PI3K/AKT signaling pathway was inhibited. Conclusion Our findings suggested that ESCs-sEVs could improve ovarian function by regulating the PI3K/AKT signaling pathway, which could provide a promising clinical therapy for POF.

Abstract Background As one of the most important transcription factor families, GRAS proteins are involved in numerous regulatory processes, especially plant growth and development. However, they have not been systematically analyzed in Brachypodium distachyon, a new model grass. Results In this study, 48 BdGRAS genes were identified. Duplicated genes account for 41.7% of them and contribute to the expansion of this gene family. 33, 39, 35 and 35 BdGRAS genes were identified by synteny with their orthologs in rice, sorghum, maize and wheat genome, respectively, indicating close relationships among these species. Based on their phylogenic relationships to GRAS genes in rice and maize, BdGRAS genes can be divided into ten subfamilies in which members of the same subfamily showed similar protein sequences, conserved motifs and gene structures, suggesting possible conserved functions. Although expression variation is high, some BdGRAS genes are tissue-specific, phytohormones- or abiotic stresses-responsive, and they may play key roles in development, signal transduction pathways and stress responses. In addition, DELLA genes BdSLR1 and BdSLRL1 were functionally characterized to play a role in plant growth via the GA signal pathway, consistent with GO annotations and KEGG pathway analyses. Conclusions Systematic analyses of BdGRAS genes indicated that members of the same subfamily may play similar roles. This was supported by the conserved functions of BdSLR1 and BdSLRL1 in GA pathway. These results laid a foundation for further functional elucidation of BdGRAS genes, especially, BdSLR1 and BdSLRL1.

Abstract Background Angiogenesis, as an endogenous repair mechanism, plays crucial roles in wound healing and tissue regeneration. However, this process is impaired in the elderly due to aging-related vascular endothelial dysfunction. This study was aimed to explore the pro-angiogenic effects of exosomes from human embryonic stem cells (ESC-Exos) in aged mice of pressure-induced ulcer model and the underlying mechanism. Methods Pressure ulcer wounds were created on the back of d-galactose-induced aging mice. ESC-Exos were locally applied onto the wound beds, with PBS as control. The effects of ESC-Exos on wound healing were analyzed by measuring wound closure rates, histological and immunofluorescence analyses. Then, the anti-aging effect of ESC-Exos on vascular endothelial cells was tested in an in vitro d-galactose-induced HUVEC senescence model. Results ESC-Exos could accelerate wound closure and enhance angiogenesis, and the senescence of vascular endothelial cells was significantly ameliorated after ESC-Exos treatment. In vitro, ESC-Exos could rejuvenate the senescence of endothelial cells and recover compromised proliferation, migratory capacity, and tube formation. This recovery was Nrf2-activation-dependent, since cotreatment with Nrf2 inhibitor Brusatol could abolish the rejuvenative effects of ESC-Exos. Further study revealed that miR-200a was highly enriched in ESC-Exos and played a crucial role in ESC-Exos-mediated rejuvenation through downregulating Keap1, which negatively regulates Nrf2 expression. Conclusions ESC-Exos ameliorate endothelial senescence by activating Nrf2 and recover aging-related angiogenic dysfunction, thereby accelerating wound healing in aged mice. ESC-Exos might be a natural nano-biomaterial for aging-related diseases therapy.

Abstract Background Previous studies revealed that urine-derived stem cells (USCs) could promote myogenesis after the impairment of the sphincter muscles. However, the effects of exosomes secreted by USCs (USCs-Exo) were not elucidated. Exosomes are nanosized membrane vesicles secreted by the cells. They have been proved to be effective in protecting against tissue injury and therapeutic in tissue repair. USCs are ideal sources of exosomes because of the noninvasive obtaining method and self-renewal abilitiy. This study aimed to show the therapeutic effects of USCs-Exo on improving stress urinary incontinence (SUI). Methods Rat SUI models were established in this study using vaginal balloon inflation, and urodynamic and histological examination were carried out after exosome application. The proliferation and differentiation of muscle satellite cells (SCs) were evaluated using EdU, Cell Counting Kit 8, immunofluorescence staining, and Western blot analysis. mRNAs and proteins related to the activation of SCs were detected by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) and Western blot analysis. Results After exosome injection, the urodynamic parameters significantly improved and the injured muscle tissue recovered well. The activation, proliferation, and differentiation of SCs were promoted. The phosphorylation of extracellular-regulated protein kinases (ERK) was enhanced. When ERK was inhibited, the promoting effect of USCs-Exo treatment disappeared. Conclusion The findings of this study elucidated the functional roles of USCs-Exo in satellite cell ERK phosphorylation and identified a novel agent for skeletal muscle regeneration, providing a basis for further exploring a cell-free correction for SUI.

Objective: To compare the effects of pulsating and static cupping on non-specific neck pain and local skin microcirculation blood perfusion, which is a pilot study. Methods: Seventy participants with non-specific neck pain were randomized to the following groups: low-frequency pulsating cupping (LF, n = 20); high-frequency pulsating cupping (HF, n = 20); static cupping (SC, n = 20), or waiting list (WL, n = 10). The LF, HF, and SC received a bilateral 10-minute cupping treatment at Jianzhongshu (SI 15). Outcomes were pain intensity (visual analog scale, VAS), functional status (Neck Disability Index, NDI), and skin blood perfusion at the SI 15, Dazhui (GV 14), and Shenzhu (GV 12) acupoint areas, measured using Laser Speckle Contrast Analysis technology. Results: Both LF and HF groups showed a significant reduction in VAS scores compared with the SC group (9.00, 95% CI 1.05–16.95, P = .027; 8.75, 95% CI 0.80–16.70, P = .031). There was no significant difference in VAS scores between the LF and HF groups (P > .05) and between NDI scores measured 3 days after intervention among the four groups (P > .05). In the SI 15 area, blood perfusion in the three treatment groups was higher than that in WL group (P

In recent years, the research of flexible sensors has become a hot topic in the field of wearable technology, attracting the attention of many researchers. However, it is still a difficult challenge to prepare low-cost and high-performance flexible sensors by a simple process. Three-dimensional spacer fabric (SF) are the ideal substrate for flexible pressure sensors due to its good compression resilience and high permeability (5747.7 mm/s, approximately 10 times that of cotton). In this paper, Thermoplastic polyurethane/Polypyrrole/Polydopamine/Space Fabric (TPU/PPy/PDA/SF) composite fabrics were prepared in a simple in-situ polymerization method by sequentially coating polydopamine (PDA) and Polypyrrole (PPy) on the surface of SF, followed by spin-coating of different polymers (thermoplastic polyurethane (TPU), polydimethylsiloxane (PDMS) and Ecoflex) on the PPy/PDA/SF surface. The results showed that the TPU/PPy/PDA/SF pressure sensors prepared by spin-coating TPU at 900 rpm at a concentration of 0.3 mol of pyrrole monomer (py) and a polymerization time of 60 min have optimum sensing performance, a wide working range (0–10 kPa), high sensitivity (97.28 kPa−1), fast response (60 ms), good cycling stability (>500 cycles), and real-time motion monitoring of different parts of the body (e.g., arms and knees). The TPU/PPy/PDA/SF piezoresistive sensor with high sensitivity on a highly permeable spacer fabric base developed in this paper has promising applications in the field of health monitoring.

Abstract Vascular dementia (VD) is one of the most common types of dementia, however, the intrinsic mechanism is unclear and there is still lack of effective medications. In this study, the VD rats exhibit a progressive cognitive impairment, as well as a time‐related increasing in hippocampal neural stem cells (H‐NSCs) senescence, lost and neurogenesis decline. Then, embryonic stem cell‐derived small extracellular vesicles (ESC‐sEVs) are intravenously injected into VD rats. ESC‐sEVs treatment significantly alleviates H‐NSCs senescence, recovers compromised proliferation and neuron differentiation capacity, and reverses cognitive impairment. By microarray analysis and RT‐qPCR it is identified that several miRNAs including miR‐17‐5p, miR‐18a‐5p, miR‐21‐5p, miR‐29a‐3p, and let‐7a‐5p, that can inhibit mTORC1 activation, exist in ESC‐sEVs. ESC‐sEVs rejuvenate H‐NSCs senescence partly by transferring these miRNAs to inhibit mTORC1 activation, promote transcription factor EB (TFEB) nuclear translocation and lysosome resumption. Taken together, these data indicate that H‐NSCs senescence cause cell depletion, neurogenesis reduction, and cognitive impairment in VD. ESC‐sEVs treatment ameliorates H‐NSCs senescence by inhibiting mTORC1 activation, and promoting TFEB nuclear translocation and lysosome resumption, thereby reversing senescence‐related neurogenesis dysfunction and cognitive impairment in VD. The application of ESC‐sEVs may be a novel cell‐free therapeutic tool for patients with VD, as well as other aging‐related diseases.

Tissue-resident stem cell senescence leads to stem cell exhaustion, which is a major cause of physiological and pathological ageing. Stem cell-derived extracellular vesicles (SC-EVs) have been reported in preclinical studies to possess therapeutic potential for diverse diseases. However, whether SC-EVs can rejuvenate senescent tissue stem cells to prevent age-related disorders still remains unknown. Here, we show that chronic application of human embryonic stem cell-derived small extracellular vesicles (hESC-sEVs) rescues the function of senescent bone marrow mesenchymal stem cells (BM-MSCs) and prevents age-related bone loss in ageing mice. Transcriptome analysis revealed that hESC-sEVs treatment upregulated the expression of genes involved in antiaging, stem cell proliferation and osteogenic differentiation in BM-MSCs. Furthermore, liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis identified 4122 proteins encapsulated in hESC-sEVs. Bioinformatics analysis predicted that the protein components in the hESCs-sEVs function in a synergistic way to induce the activation of several canonical signalling pathways, including Wnt, Sirtuin, AMPK, PTEN signalling, which results in the upregulation of antiaging genes in BM-MSCs and then the recovery of senescent BM-MSCs function. Collectively, our findings reveal the effect of hESC-sEVs in reversing BM-MSCs senescence and age-related osteogenic dysfunction, thereby preventing age-related bone loss. Because hESC-sEVs could alleviate senescence of tissue-resident stem cells, they might be promising therapeutic candidates for age-related diseases.

Background/Aims: Our previous studies have shown that human urine-derived stem cells (USCs) have great potential as a cell source for cytotherapy and tissue engineering and that extracellular vesicles (EVs) secreted by USCs (USCs-EVs) can prevent diabetes-induced kidney injury in an animal model. The present study was designed to evaluate the effects of USCs-EVs on ischemia repair. Methods: USCs-EVs were isolated and purified by a battery of centrifugation and filtration steps. The USCs-EVs were then characterized by transmission electron microscopy, western blot and tunable resistive pulse sensing techniques. After intramuscularly transplanting USCs-EVs into an ischemic mouse hind-limb, we observed the therapeutic effects of USCs-EVs on perfusion by laser doppler perfusion imaging, angiogenesis and muscle regeneration by histology and immunohistochemistry techniques over 21 days. We subsequently tested whether USCs-EVs can induce the proliferation of a human microvascular endothelial cell line HMEC-1 and a mouse myoblast cell line C2C12 by cell counting kit 8 assay in vitro. Meanwhile, the potential growth factors in the USCs-EVs and supernatants of the USCs cultures were detected by enzyme-linked immunosorbent assay. Results: The USCs-EVs were spherical vesicles with a diameter of 30–150 nm and expressed exosomal markers, such as CD9, CD63 and Tsg101. Ischemic limb perfusion and function were markedly increased in the hind-limb ischemia (HLI) model after USCs-EVs administration. Moreover, angiogenesis and muscle regeneration levels were significantly higher in the USCs-EVs treatment group than in the PBS group. The in vitro experiments showed that USCs-EVs facilitated HMEC-1 and C2C12 cell proliferation in a dose-dependent manner. Conclusions: These results revealed for the first time that USCs-EVs efficiently attenuate severe hind-limb ischemic injury and represent a novel therapy for HLI.

Abstract Background Breast cancer survivors are at high risk for fracture due to cancer treatment-induced bone loss, however, data is scarce regarding the scope of this problem from an epidemiologic and health services perspective among Chinese women with breast cancer. Methods We designed a cross-sectional study comparing prevalence of vertebral fractures among age- and BMI-matched women from two cohorts. Women in the Breast Cancer Survivors cohort were enrolled from a large cancer hospital in Beijing. Eligibility criteria included age 50–70 years, initiation of treatment for breast cancer at least 5 years prior to enrollment, and no history of metabolic bone disease or bone metastases. Data collected included sociodemographic characteristics; fracture-related risk factors, screening and preventive measures; breast cancer history; and thoracolumbar x-ray. The matched comparator group was selected from participants enrolled in the Peking Vertebral Fracture Study, an independent cohort of healthy community-dwelling postmenopausal women from Beijing. Results Two hundred breast cancer survivors were enrolled (mean age 57.5 ± 4.9 years), and compared with 200 matched healthy women. Twenty-two (11%) vertebral fractures were identified among breast cancer survivors compared with 7 (3.5%) vertebral fractures in the comparison group, yielding an adjusted odds ratio for vertebral fracture of 4.16 (95%CI 1.69–10.21, p

Abstract Background Human induced pluripotent stem cells (iPSCs) have been verified as a powerful cell model for the study of pathogenesis in hereditary disease. Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations of PKD or non-PKD genes. The pathogenesis of ADPKD remains unexplored because of the lack of a true human cell model. Methods Six ADPKD patients and four healthy individuals were recruited as donors of somatic cells from a Chinese ADPKD family without mutations of the PKD genes but carrying SAMSN1 gene deletion. The ADPKD-iPSCs were generated from somatic cells and were induced into kidney-like cells (KLCs) by a novel three-step method involving cytokines and renal epithelium growth medium. Furthermore, we analyzed functional properties of these KLCs by water transportation and albumin absorption assays. Results We successfully generated iPSCs from ADPKD patients and differentiated them into KLCs that showed morphological and functional characteristics of human kidney cells. Further, we also found that ADPKD-iPSC-KLCs had a significantly higher rate of apoptosis and a significantly lower capacity for water transportation and albumin absorption compared to healthy sibling-derived differentiated KLCs. Furthermore, knockdown of SAMSN1 in control iPSCs may attenuate differentiation and/or function of KLCs. Conclusions These data show that we have created the first iPSCs established from ADPKD patients without mutations in the PKD genes, and suggest that the deletion mutation of SAMSN1 might be involved in the differentiation and/or function of KLCs. ADPKD-iPSC-KLCs can be used as a versatile model system for the study of kidney disease.

Abstract Background As a superfamily of transcription factors (TFs), the basic helix-loop-helix (bHLH) proteins have been characterized functionally in many plants with a vital role in the regulation of diverse biological processes including growth, development, response to various stresses, and so on. However, no systemic analysis of the bHLH TFs has been reported in Brachypodium distachyon, an emerging model plant in Poaceae. Results A total of 146 bHLH TFs were identified in the Brachypodium distachyon genome and classified into 24 subfamilies. BdbHLHs in the same subfamily share similar protein motifs and gene structures. Gene duplication events showed a close relationship to rice, maize and sorghum, and segment duplications might play a key role in the expansion of this gene family. The amino acid sequence of the bHLH domains were quite conservative, especially Leu-27 and Leu-54. Based on the predicted binding activities, the BdbHLHs were divided into DNA binding and non-DNA binding types. According to the gene ontology (GO) analysis, BdbHLHs were speculated to function in homodimer or heterodimer manner. By integrating the available high throughput data in public database and results of quantitative RT-PCR, we found the expression profiles of BdbHLHs were different, implying their differentiated functions. Conclusion One hundred fourty-six BdbHLHs were identified and their conserved domains, sequence features, phylogenetic relationship, chromosomal distribution, GO annotations, gene structures, gene duplication and expression profiles were investigated. Our findings lay a foundation for further evolutionary and functional elucidation of BdbHLH genes.

To enhance the decay resistance of bamboo oriented strand board (OSB) products, the strands were dipped in solutions of alkaline copper quat (ACQ) and copper azole (CA) and bonded with phenol formaldehyde resin into two types of OSB panels, i.e., panels with 100% treated strands and those with treated strands only in the face layers. The results indicated that the decay resistance of treated panels was effectively enhanced. The physical and mechanical properties of all treated panels exceeded the requirements specified for category OSB/4 or OSB/3 in the standard LY/T 1580-2010. Statistical data analysis showed that pretreatment with ACQ and CA did not have detrimental effects on the overall physical and mechanical properties of panels at the loading levels investigated in this study. Panels with pretreated strands only in the face layers had strong decay resistance and comparable overall properties as those with 100% treated strands. The results suggest that pretreatment is a promising way to introduce waterborne ACQ and CA to protect bamboo OSB.

Aircraft recognition has great application value, but aircraft in remote sensing images have some problems such as low resolution, poor contrasts, poor sharpness, and lack of details caused by the vertical view, which make the aircraft recognition very difficult. Especially when there are many kinds of aircraft and the differences between aircraft are subtle, the fine-grained recognition of aircraft is more challenging. In this paper, we propose a non-locally enhanced feature fusion network(NLFFNet) and attempt to make full use of the features from discriminative parts of aircraft. First, according to the long-distance self-correlation in aircraft images, we adopt non-locally enhanced operation and guide the network to pay more attention to the discriminating areas and enhance the features beneficial to classification. Second, we propose a part-level feature fusion mechanism(PFF), which crops 5 parts of the aircraft on the shared feature maps, then extracts the subtle features inside the parts through the part full connection layer(PFC) and fuses the features of these parts together through the combined full connection layer(CFC). In addition, by adopting the improved loss function, we can enhance the weight of hard examples in the loss function meanwhile reducing the weight of excessively hard examples, which improves the overall recognition ability of the network. The dataset includes 47 categories of aircraft, including many aircraft of the same family with slight differences in appearance, and our method can achieve 89.12% accuracy on the test dataset, which proves the effectiveness of our method.

RhoA is a critical signaling molecule regulating a variety of cellular processes, such as cytoskeletal organization, adhesion, and apoptosis. It is recently considered responsive to reactive oxygen species (ROS). Nevertheless, how RhoA regulates anoikis, a detachment-initiated apoptosis, and how this regulation is affected by ROS are not clear. The present study investigated the role of RhoA in apoptosis/anoikis in gastric cancer cells and the changes of RhoA and anoikis under oxidative stress. Immunohistochemistry showed that RhoA expression was upregulated in the primary gastric carcinoma compared with normal gastric mucosa. Overactivation of RhoA by transfection with the V14RhoA mutant prevented gastric cancer line SGC-7901 cells from arsenic-induced apoptosis and conferred anoikis resistance through, at least in part, promoting formations of F-actin fibers and focal adhesion. Oxidative stress caused by emodin, an ROS producer, in combination with arsenic trioxide (ATO) led to RhoA inactivation that triggered structural disruption of focal adhesion complex and eventually resulted in anoikis, and these effects could be partially reversed by antioxidant N-acetylcysteine (NAC). In conclusion, activation of RhoA is required for the maintenance of anoikis resistance phenotype of gastric cancer cells, and oxidative stress might be a therapeutic strategy for the inhibition of RhoA in cancer cells.

Bone tissue engineering requires highly proliferative stem cells that are easy to isolate. Human urine stem cells (USCs) are abundant and can be easily harvested without using an invasive procedure. In addition, in our previous studies, USCs have been proved to be able to differentiate into osteoblasts, chondrocytes, and adipocytes. Therefore, USCs may have great potential and advantages to be applied as a cell source for tissue engineering. However, there are no published studies that describe the interactions between USCs and biomaterials and applications of USCs for bone tissue engineering. Therefore, the objective of the present study was to evaluate the interactions between USCs with a typical bone tissue engineering scaffold, beta-Tricalcium Phosphate (β-TCP), and to determine whether the USCs seeded onto β-TCP scaffold can promote bone regeneration in a segmental femoral defect of rats. Primary USCs were isolated from urine and seeded on β-TCP scaffolds. Results showed that USCs remained viable and proliferated within β-TCP. The osteogenic differentiation of USCs within the scaffolds was demonstrated by increased alkaline phosphatase activity and calcium content. Furthermore, β-TCP with adherent USCs (USCs/β-TCP) were implanted in a 6-mm critical size femoral defect of rats for 12 weeks. Bone regeneration was determined using X-ray, micro-CT, and histologic analyses. Results further demonstrated that USCs in the scaffolds could enhance new bone formation, which spanned bone defects in 5 out of 11 rats while β-TCP scaffold alone induced modest bone formation. The current study indicated that the USCs can be used as a cell source for bone tissue engineering as they are compatible with bone tissue engineering scaffolds and can stimulate the regeneration of bone in a critical size bone defect.

The discovery of induced pluripotent stem cells (iPSCs) rendered the reprogramming of terminally differentiated cells to primary stem cells with pluripotency possible and provided potential for the regeneration and restoration of cartilage defect. Chondrogenic differentiation of iPSCs is crucial for their application in cartilage tissue engineering. In this study we investigated the effect of 3D nanofibrous scaffolds on the chondrogenesis of iPSCs and articular cartilage defect restoration. Super-hydrophilic and durable mechanic polycaprolactone (PCL)/gelatin scaffolds were fabricated using two separate electrospinning processes. The morphological structure and mechanical properties of the scaffolds were characterized. The chondrogenesis of the iPSCs in vitro and the restoration of the cartilage defect was investigated using scanning electron microscopy (SEM), the Cell Counting Kit-8 (CCK-8), histological observation, RT-qPCR, and western blot analysis. iPSCs on the scaffolds expressed higher levels of chondrogenic markers than the control group. In an animal model, cartilage defects implanted with the scaffold-cell complex exhibited an enhanced gross appearance and histological improvements, higher cartilage-specific gene expression and protein levels, as well as subchondral bone regeneration. Therefore, we showed scaffolds with a 3D nanofibrous structure enhanced the chondrogenesis of iPSCs and that iPSC-containing scaffolds improved the restoration of cartilage defects to a greater degree than did scaffolds alone in vivo.

In this experiment, we established an animal model of gestational diabetes mellitus rats using streptozotocin. Using the rat model of GDM, the pregnant rats in 1-19d were divided into three groups: (1) Zuogui Wan gestational diabetes mellitus group (group I, n=12), (2) gestational diabetes mellitus rats as the control group (group II, n=11), and (3) rats of normal pregnancy group (group III, n=11). Compared with gestational diabetes mellitus rats as the control group, Zuogui Wan can change the indexes of fasting blood glucose, body weight, total cholesterol, insulin, and metabolism cage index significantly in Zuogui Wan gestational diabetes mellitus group. We can conclude that Zuogui Wan has the therapeutic effect on gestational diabetes mellitus.