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Abstract Background The high complexity of systemic autoimmune diseases (SADs) has hindered precise management. This study aims to investigate heterogeneity in SADs. Methods We applied a joint cluster analysis, which jointed multiple correspondence analysis and k-means, to immunomarkers and measured the heterogeneity of clusters by examining differences in immunomarkers and clinical manifestations. The electronic health records of patients who received an antinuclear antibody test and were diagnosed with SADs, namely systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and Sjögren’s syndrome (SS), were retrieved between 2001 and 2016 from hospitals in Taiwan. Results With distinctive patterns of immunomarkers, a total of 11,923 patients with the three SADs were grouped into six clusters. None of the clusters was composed only of a single SAD, and these clusters demonstrated considerable differences in clinical manifestation. Both patients with SLE and SS had a more dispersed distribution in the six clusters. Among patients with SLE, the occurrence of renal compromise was higher in Clusters 3 and 6 (52% and 51%) than in the other clusters (p

Background and purpose: Acupuncture exerts an anti-inflammatory effect and is recommended by the World Health Organization as a complementary therapy for stroke. This study investigated the improvement in neurological function outcome in acute-stage intervention of acute ischemic stroke (AIS), and the anti-inflammatory effect of early acupuncture. Methods: Fifty patients with AIS were randomly assigned to either a control group (CG, 25 patients, received sham acupuncture) or treatment group (TG, 25 patients, received acupuncture treatment). Acupuncture intervention was administered twice a week for a total of 8 sessions over 4 consecutive weeks. The primary outcome was the changes in the National Institutes of Health Stroke Scale (NIHSS), modified Rankin Scale (mRS), and Barthel Index (BI) scores. The secondary outcome was the changes in serum inflammation-related biomarker levels.(ANAIS trial) Results: A total of 35 patients (18 patients in the CG and 17 patients in the TG) completed the trial. The reduction in NIHSS scores was greater in the TG than in the CG between V2 (second assessment administered after acupuncture intervention) and V1 (first assessment administered before acupuncture intervention; 4.33 ± 1.91 vs. 2.68 ± 1.42, p = 0.005) and between V3 (third assessment administered 28 days after last acupuncture intervention) and V1 (6.00 ± 2.53 vs. 3.83 ± 2.31, p = 0.012). The increase in BI scores was greater in the TG than in the CG between V2 and V1 (28.89 ± 15.39 vs. 14.21 ± 19.38, p = 0.016) and between V3 and V1 (39.41 ± 20.98 vs. 25.00 ± 18.47, p = 0.038). Among participants with high inflammation, the increase in serum IL-12p70 level between V2 and V1 was greater in the TG than in the CG (0.20 ± 0.19 vs. −0.14 ± 0.30, pg/mL p = 0.006). Conclusions: Acupuncture improved the neurological function of patients with AIS, and the relationship between acupuncture improving neurological function and anti-inflammatory effect needs further study. In addition, studies with larger sample sizes and longer follow-ups as well as multicenter clinical trials are expected in the future.

Aims: Several artificial bone grafts have been developed but fail to achieve anticipated osteogenesis due to their insufficient neovascularization capacity and periosteum support. This study aimed to develop a vascularized bone-periosteum construct (VBPC) to provide better angiogenesis and osteogenesis for bone regeneration. Methods: A total of 24 male New Zealand white rabbits were divided into four groups according to the experimental materials. Allogenic adipose-derived mesenchymal stem cells (AMSCs) were cultured and seeded evenly in the collagen/chitosan sheet to form cell sheet as periosteum. Simultaneously, allogenic AMSCs were seeded onto alginate beads and were cultured to differentiate to endothelial-like cells to form vascularized bone construct (VBC). The cell sheet was wrapped onto VBC to create a vascularized bone-periosteum construct (VBPC). Four different experimental materials – acellular construct, VBC, non-vascularized bone-periosteum construct, and VBPC – were then implanted in bilateral L4-L5 intertransverse space. At 12 weeks post-surgery, the bone-forming capacities were determined by CT, biomechanical testing, histology, and immunohistochemistry staining analyses. Results: At 12 weeks, the VBPC group significantly increased new bone formation volume compared with the other groups. Biomechanical testing demonstrated higher torque strength in the VBPC group. Notably, the haematoxylin and eosin, Masson’s trichrome, and immunohistochemistry-stained histological results revealed that VBPC promoted neovascularization and new bone formation in the spine fusion areas. Conclusion: The tissue-engineered VBPC showed great capability in promoting angiogenesis and osteogenesis in vivo. It may provide a novel approach to create a superior blood supply and nutritional environment to overcome the deficits of current artificial bone graft substitutes. Cite this article: Bone Joint Res 2023;12(12):722–733.

Abstract Backgrounds Bone marrow stem cell can differentiate to osteoblast by growth factors, pulsed low-intensity ultrasound and electric magnetic field. In the research, bone marrow stem cells were cultured; bone marrow stem cells in culture can be stimulated by platelet-rich plasma and electric field. Methods The culture well of the co-cultivation device has a radius of 7.5 mm and a depth of 7 mm. It is divided into two sub-chambers separated by a 3 mm high and 1 mm wide barrier. The bone marrow stem cells were seeded at a density of 2 × 104 cells and the medium volume was 120μl. Platelet-rich plasma (PRP) or platelet-poor plasma (PPP) was added to the other sub-chamber at a volume of 10μl. The bone marrow stem cells were subjected to different electric fields (0 ~ 1 V/cm) at a frequency of 70 kHz for 60 min. Results The highest osteogenic capacity of bone marrow stem cells was achieved by addition of PRP to electric field stimulation (0.25 V/cm) resulted in a proliferation rate of 599.78%. In electric field stimulation (0.75 V/cm) with PPP, the proliferation rate was only 10.46%. Conclusions Bone marrow stem cell with PRP in the co-culture device combined with electric field at 0.25 V/cm strength significantly promoted the growth of bone marrow stem cells.

Notch signaling is a conserved pathway crucial for nervous system development. Disruptions in this pathway are linked to neurodevelopmental disorders, neurodegenerative diseases, and brain tumors. Hairy/E(spl) (HES) genes, major downstream targets of Notch, are commonly used as markers for Notch activation. However, these genes can be activated, inhibited, or function independently of Notch signaling, and their response to Notch disruption varies across tissues and developmental stages. MIB1/Mib1 is an E3 ubiquitin ligase that enables Notch receptor activation by processing ligands like Delta and Serrate. We investigated Notch signaling disruption using the zebrafish Mib1 mutant line, mib1ta52b, focusing on changes in the expression of Hairy/E(spl) (her) genes. Our findings reveal significant variability in her gene expression across different neural cell types, regions, and developmental stages following Notch disruption. This variability questions the reliability of Hairy/E(spl) genes as universal markers for Notch activation, as their response is highly context-dependent. This study highlights the complex and context-specific nature of Notch signaling regulation. It underscores the need for a nuanced approach when using Hairy/E(spl) genes as markers for Notch activity. Additionally, it provides new insights into Mib1’s role in Notch signaling, contributing to a better understanding of its involvement in Notch signaling-related disorders.

Background: Hip fracture is a common disease in the elderly. Among these patients, surgical intervention for hip fracture should be carefully considered because of old age and multiple comorbidities. There are still insufficient comparisons between nonagenarian patients treated with surgery and those treated non-surgically. We studied hip fracture nonagenarian patients to compare the different outcomes between surgical and non-surgical treatments. Materials and methods: Nonagenarian patients visiting the emergency department with hip fractures between March 2010 and December 2020 were identified. Overall survival was estimated using multivariate Cox proportional hazards models. The mortality rates, the length of hospital stay, complication and readmission rates were also recorded. Results: A total of 173 patients who underwent surgery and 32 who received conservative treatments were included. The median survival time was 58.47 months in the OP group, which was significantly higher than the 24.28 months in the non-OP group. After adjusting for covariates, including age, sex, Charlson Comorbidity Index (CCI), injury severity score, and fracture type, the risk of death was reduced by surgery (hazard ratio [HR] = 0.427; 95 % confidence interval [CI]: 0.207–0.882; p = 0.021). CCI was also an independent risk factor for poor survival rate (HR = 1.3; 95 % CI: 1.115–1.515; p = 0.001). After adjusting for several factors, surgery within 48 h improved overall survival (HR: 2.518; 95 % CI: 1.299–4.879; p = 0.006) in operative group. Conclusion: Our study suggests that surgical treatment may provide better survival for nonagenarian patients with hip fractures than non-operation, especially patients with less concurrent comorbidities.

Key message Congenital anomalies of the large airways are occasionally asymptomatic and are incidentally discovered through radiography, often using computed tomography. Bronchoscopy can aid in the direct visual examination of the large airway abnormalities detected on radiography.

Frozen-thawed embryo transfer (FET) has been a viable alternative to fresh embryo transfer in recent years because of the improvement in vitrification methods. Laboratory-based studies indicate that complex molecular and morphological changes in endometrium during the window of implantation after exogenous hormones with controlled ovarian stimulation may alter the interaction between the embryo and endometrium, leading to a decreased implantation potential. Based on the results obtained from randomized controlled studies, increased pregnancy rates and better perinatal outcomes have been reported following FET. Compared to fresh embryo transfer, fewer preterm deliveries, and reduced incidence of ovarian hyperstimulation syndrome were found after FETs, yet there is a trend of increased pregnancy-related hypertensive diseases in women receiving FET. Despite the increased application of FET, the search for the most optimal priming protocol for the endometrium is still undergoing. Three available FET protocols have been proposed to prepare the endometrium: i) natural cycle (true natural cycle and modified natural cycle) ii) artificial cycle (AC) or hormone replacement treatment cycle iii) mild ovarian stimulation (mild-OS) cycle. Emerging evidence suggests that the optimal timing for FET using warmed blastocyst transfer is the LH surge+6 day, hCG administration+7 day, and the progesterone administration+6 day in the true natural cycle, modified natural cycle, and AC protocol, respectively. Although still controversial, better clinical pregnancy rates and live birth rates have been reported using the natural cycle (true natural cycle/modified natural cycle) compared with the AC protocol. Additionally, a higher early pregnancy loss rate and an increased incidence of gestational hypertension have been found in FETs using the AC protocol because of the lack of a corpus luteum. Although the common clinical practice is to employ luteal phase support (LPS) in natural cycles and mild-OS cycles for FET, the requirement for LPS in these protocols remains equivocal. Recent findings obtained from RCTs do not support the routine application of endometrial receptivity testing to optimize the timing of FET. More RCTs with rigorous methodology are needed to compare different protocols to prime the endometrium for FET, focusing not only on live birth rate, but also on maternal, obstetrical, and neonatal outcomes.

Background: Surgical intervention is important for acute appendicitis. Perforated appendicitis leads to more complications and is more difficult to manage. Studies differentiating ruptured from nonruptured appendicitis are controversial. Aim: Early identification of perforated appendicitis is important for first-line medical practitioners. Our study aimed to provide more indicators to evaluate the risk of perforation before surgery. Methods: This retrospective study included 116 patients who underwent an appendectomy at the Tri-Service General Hospital between 2014 and 2018 with pathological diagnosis. Based on the surgical records, the patients were classified into two groups: simple (nonperforation) and complicated (perforation). Results: The patients (68 boys, 48 girls) were all under 18 years of age. Simple appendicitis was diagnosed in 87 patients and perforated appendicitis in 29 patients. Analysis revealed significant differences in blood C-reactive protein (CRP) levels between the two groups. The highest Youden's index was at a CRP level of 1.755 mg/dL. Positive findings were obtained for suppurative change as well as chylous ascites. Conclusion: Elevated CRP level was found to be a useful biomarker in predicting perforated appendicitis, excluding other general factors in pediatric patients. Suppurative change and chylous ascites could serve as indicators for the possibility of perforation during surgery.

Background/Purpose: Hip fractures are associated with physical dysfunction, and poor quality of life in the elderly. Post-acute care (PAC) would facilitate functional recovery in patients with hip fractures after surgeries. Taiwan has proposed a nationwide PAC program for hip fractures since 2017, but little has been known about its effectiveness. Therefore, this study aimed to evaluate the efficacy and cost-effectiveness of the PAC program for hip fracture patients in Taiwan. Methods: This was a prospective study. Patients aged ≥ 65 years with hip fractures after surgeries were recruited and divided into home-based, hospital-based, and control groups. Outcome measures included pain, physical function (sit-to-stand test, Barthel Index [BI], and Harris hip score [HHS]), and quality of life (EuroQol instrument [EQ-5D]). Direct medical and non-medical costs were recorded. Cost-effectiveness ratio (CER) was calculated as the amount of New Taiwanese Dollars (NTDs) paid per BI and EQ-5D unit improvement. Results: Forty-one patients participated in this study, with 17, 12, and 12 in the home-based, hospital-based, and control groups, respectively. The home-based group showed significant improvements in BI and HHS compared to the controls (p = 0.018 and p = 0.029, respectively). The hospital-based group demonstrated significant improvement in EQ-5D compared to the controls (p = 0.015). The home-based PAC program demonstrated the best CER for BI (NTD 554) and EQ-5D (NTD 41948). Conclusion: Both PAC programs would significantly improve the physical function and quality of life in patients with hip fractures. However, the home-based PAC provided the best CER for BI and EQ-5D.

Early pregnancy is a complex and well-orchestrated differentiation process that involves all the cellular elements of the fetal-maternal interface. Aberrant trophoblast-decidual interactions can lead to miscarriage and disorders that occur later in pregnancy, including preeclampsia, intrauterine fetal growth restriction, and preterm labor. A great deal of research on the regulation of implantation and placentation has been performed in a wide range of species. However, there is significant species variation regarding trophoblast differentiation as well as decidual-specific gene expression and regulation. Most of the relevant information has been obtained from studies using mouse models. A comprehensive understanding of the physiology and pathology of human implantation and placentation has only recently been obtained because of emerging advanced technologies. With the derivation of human trophoblast stem cells, 3D-organoid cultures, and single-cell analyses of differentiated cells, cell type-specific transcript profiles and functions were generated, and each exhibited a unique signature. Additionally, through integrative transcriptomic information, researchers can uncover the cellular dysfunction of embryonic and placental cells in peri-implantation embryos and the early pathological placenta. In fact, the clinical utility of fetal-maternal cellular trafficking has been applied for the noninvasive prenatal diagnosis of aneuploidies and the prediction of pregnancy complications. Furthermore, recent studies have proposed a viable path toward the development of therapeutic strategies targeting placenta-enriched molecules for placental dysfunction and diseases.

Strongyloidiasis is caused by infection with the helminth, Strongyloides stercoralis. The life cycle of S. stercoralis in the host could last decades owing to autoinfection. Most symptoms of strongyloidiasis are nonspecific and may involve the skin, respiratory tract, or gastrointestinal tract. Severe anemia is a less frequently reported symptom of strongyloidiasis. The prevalence of strongyloidiasis is low in Taiwan, and sporadic cases have been previously reported. We present a case of strongyloidiasis in an 82-year-old woman diagnosed by microscopic examination of stool, presenting with severe normocytic anemia. After treatment with the antiparasitic medication, ivermectin, and blood transfusion, her anemia improved. Therefore, parasite infection should be considered in immunosuppressed patients despite atypical laboratory results. Physicians should be aware of the rare incidence of strongyloidiasis in Taiwan and consider it as a possible differential diagnosis for anemia.

Secreted by the anterior pituitary gland, growth hormone (GH) is a peptide that plays a critical role in regulating cell growth, development, and metabolism in multiple targeted tissues. Studies have shown that GH and its functional receptor are also expressed in the female reproductive system, including the ovaries and uterus. The experimental data suggest putative roles for GH and insulin-like growth factor 1 (IGF-1, induced by GH activity) signaling in the direct control of multiple reproductive functions, including activation of primordial follicles, folliculogenesis, ovarian steroidogenesis, oocyte maturation, and embryo implantation. In addition, GH enhances granulosa cell responsiveness to gonadotropin by upregulating the expression of gonadotropin receptors (follicle-stimulating hormone receptor and luteinizing hormone receptor), indicating crosstalk between this ovarian regulator and the endocrine signaling system. Notably, natural gene mutation of GH and the age-related decline in GH levels may have a detrimental effect on female reproductive function, leading to several reproductive pathologies, such as diminished ovarian reserve, poor ovarian response during assisted reproductive technology (ART), and implantation failure. Association studies using clinical samples showed that mature GH peptide is present in human follicular fluid, and the concentration of GH in this fluid is positively correlated with oocyte quality and the subsequent embryo morphology and cleavage rate. Furthermore, the results obtained from animal experiments and human samples indicate that supplementation with GH in the in vitro culture system increases steroid hormone production, prevents cell apoptosis, and enhances oocyte maturation and embryo quality. The uterine endometrium is another GH target site, as GH promotes endometrial receptivity and pregnancy by facilitating the implantation process, and the targeted depletion of GH receptors in mice results in fewer uterine implantation sites. Although still controversial, the administration of GH during ovarian stimulation alleviates age-related decreases in ART efficiency, including the number of oocytes retrieved, fertilization rate, embryo quality, implantation rate, pregnancy rate, and live birth rate, especially in patients with poor ovarian response and recurrent implantation failure.

Background On the basis of molecular dating, Pleistocene glaciations have been proposed as the major driving force of biota speciation in the Palearctic and the pre-Quaternary origin of Amazonian taxa. However, the major driving factors in East Asia remain unclear. All 16 saturniine species inhabiting Taiwan with congeners of populations, subspecies, or species in East Asia constitute research objects for addressing the mode of speciation because of the repeated formation and disappearance of a landbridge from the Asian mainland to Taiwan during glacial cycles. Methods The genetic divergences of mitochondrial cytochrome c oxidase subunit I (COI) and 16S rDNA and the nuclear 28S rDNA of the saturniine species from Taiwan and the Asian mainland were assessed to determine the monophyly of each genus and species of Saturniinae. Moreover, 519 saturniine COI sequences of 114 taxa from adjacent East and Southeast Asian populations and closely related species were retrieved from GenBank and analyzed. The differentiation timing and possible origination of the insular saturniines were elucidated based on phylogenetic relationships, haplotype networks, and lineage calibrations. Results Approximately 90% of intraspecific COI divergence was

Bone morphogenetic protein (BMP) signaling regulates neural induction, neuronal specification, and neuronal differentiation. However, the role of BMP signaling in neural progenitors remains unclear. This is because interruption of BMP signaling before or during neural induction causes severe effects on subsequent neural developmental processes. To examine the role of BMP signaling in the development of neural progenitors in zebrafish, we bypassed the effect of BMP signaling on neural induction and suppressed BMP signaling at different time points during gastrulation using a temporally controlled transgenic line carrying a dominant-negative form of Bmp receptor type 1aa and a chemical inhibitor of BMP signaling, DMH1. Inhibiting BMP signaling from 8 hpf could bypass BMP regulation on neural induction, induce the number of proliferating neural progenitors, and reduce the number of neuronal precursors. Inhibiting BMP signaling upregulates the expression of the Notch downstream gene hairy/E(spl)-related 2 (her2). Inhibiting Notch signaling or knocking down the Her2 function reduced neural progenitor proliferation, whereas inactivating BMP signaling in Notch-Her2 deficient background restored the number of proliferating neural progenitors. These results reveal the time window for the proliferation of neural progenitors during zebrafish development and a fine balance between BMP and Notch signaling in regulating the proliferation of neural progenitor cells.

Summary: Lymphoma is a group of blood cancers that develop from the immune system, and one of the main risk factors is associated with exposure to environmental chemicals. Bisphenol A (BPA) is a common chemical used in the manufacture of materials in polycarbonate and epoxy plastic products and can interfere with the immune system. BPA is considered to possibly induce lymphoma development by affecting the immune system, but its potential mechanisms have not been well established. This study performed a gene-network analysis of microarray data sets in human lymphoma tissues as well as in human cells with BPA exposure to explore module genes and construct the potential pathway for lymphomagenesis in response to BPA. This study provided evidence that BPA exposure resulted in disrupted cell cycle and DNA damage by activating CTNNB1, the initiator of the aberrant constructed CTNNB1-NFKB1-AR-IGF1-TWIST1 pathway, which may potentially lead to lymphomagenesis.

In addition to regulating immune responses by producing antibodies that confer humoral immunity, B cells can also affect these responses by producing cytokines. How B cells participate in the clearance of pathogenic infections via functions other than the production of pathogen-specific antibodies is still largely unknown. Marginal zone (MZ) B cells can quickly respond to bacterial invasion by providing the initial round of antibodies. After a bloodborne bacterial infection, neutrophils promptly migrate to the MZ. However, the mechanisms regulating neutrophil accumulation in the MZ during the initial phase of infection also remain obscure. Here, we found that MZ B cell-deficient mice are more susceptible to systemic Staphylococcus aureus (S. aureus) infection compared with wildtype mice. The expression levels of interleukin (IL)-6 and CXCL1/CXCL2 in MZ B cells increased significantly in mice at 3–4 h after infection with S. aureus, then decreased at 24 h post-infection. After systemic S. aureus infection, splenic neutrophils express increased CXCR2 levels. Our results from confocal microscopy imaging of thick-section staining demonstrate that neutrophils in wildtype mice form cell clusters and are in close contact with MZ B cells at 3 h post-infection. This neutrophil cluster formation shortly after infection was diminished in both MZ B cell-deficient mice and IL-6-deficient mice. Blocking the action of CXCL1/CXCL2 by injecting anti-CXCL1 and anti-CXCL2 antibodies 1 h before S. aureus infection significantly suppressed the recruitment of neutrophils to the MZ at 3 h post-infection. Compared with peptidoglycan stimulation alone, peptidoglycan stimulation with neutrophil co-culture further enhanced MZ B-cell activation and differentiation. Using a Förster resonance energy transfer by fluorescence lifetime imaging (FLIM-FRET) analysis, we observed evidence of a direct interaction between neutrophils and MZ B cells after peptidoglycan stimulation. Furthermore, neutrophil depletion in mice resulted in a reduced production of S. aureus-specific immunoglobulin (Ig)M at 24 h post-infection. Together, our results demonstrate that MZ B cells regulate the rapid neutrophil swarming into the spleen during the early phase of systemic S. aureus infection. Interaction with neutrophils assists MZ B cells with their differentiation into IgM-secreting cells and contributes to the clearance of systemic bacterial infections.

Abstract Terminally differentiated B cell, the plasma cell, is the sole cell type capable of producing antibodies in our body. Over the past 30 years, the identification of many key molecules controlling B cell activation and differentiation has elucidated the molecular pathways for generating antibody-producing plasma cells. Several types of regulation modulating the functions of the important key molecules in B cell activation and differentiation add other layers of complexity in shaping B cell responses following antigen exposure in the absence or presence of T cell help. Further understanding of the mechanisms contributing to the proper activation and differentiation of B cells into antibody-secreting plasma cells may enable us to develop new strategies for managing antibody humoral responses during health and disease. Herein, we reviewed the effect of different types of regulation, including transcriptional regulation, post-transcriptional regulation and epigenetic regulation, on B cell activation, and on mounting memory B cell and antibody responses. We also discussed the link between the dysregulation of the abovementioned regulatory mechanisms and B cell-related disorders.

Pain and itch are both important manifestations of various disorders, such as herpes zoster, atopic dermatitis, and psoriasis. Growing evidence suggests that both sensations have shared mediators, overlapping neural circuitry, and similarities in sensitization processes. In fact, pain and itch coexist in some disorders. Determining pharmaceutical agents and targets for treating pain and itch concurrently is of scientific and clinical relevance. Here we review the neurobiology of pain and itch and discuss the pharmaceutical targets as well as novel compounds effective for the concurrent treatment of these sensations.

Vernonia amygdalina (VA) has been reported to have antioxidant potential; however, its DNA protection and anti-inflammatory properties remain unclear. We aimed to investigate whether aqueous (WEVAL) and alcoholic (EEVAL) VA extracts exert similar antioxidant, DNA protection and anti-inflammatory effects and attempted to explore the mechanism underlying the anti-inflammatory effects. These results demonstrated that WEVAL had greater polyphenolic and flavonoid contents, as well as a stronger reducing power, DPPH radical scavenging and DNA protective activity. Moreover, both extracts reduced lipopolysaccharide (LPS)-induced expression of COX-II, iNOS, pro-inflammatory factors, including NO, TNF-α, IL-1β, and IL-10. Compared with WEVAL, EEVAL was a more potent inflammatory inhibitor. Both extracts similarly inhibited LPS-induced MAPK (p38) and NF-κB expression. Our findings indicate that WEVAL and EEVAL have diverse antioxidant and anti-inflammatory effects. WEVAL had a stronger antioxidant and DNA protection activity; contrastingly, EEVAL had a stronger anti-inflammatory ability. The anti-inflammatory activity involves reduced pro-inflammatory cytokines through NF-κB down-regulation and MAPK inhibition. These results demonstrated that production of WEVAL and EEVAL from VA leaves may provide a safe and efficacious source of pharmaceutical applications, with antioxidant, DNA protective and anti-inflammation activities.

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Mutations in the FHL2 (Four and a half LIM domains protein 2) gene are associated with cardiomyopathy in patients. Here, we generated two homozygous knockout lines using CRISPR/Cas9-mediated ablation in a human embryonic stem cell (hESC) WA09 line. These knockout lines exhibit a normal karyotype without expressing FHL2 protein, while maintaining pluripotency and differentiation properties. These isogenic mutation lines will be provided as a disease model for cardiomyopathy studies and drug screening.

The manufacture and properties of waterborne UV-cured coatings (WUV coatings) by acetone process based on urushiol for wood finishing were investigated. Firstly, epoxide urushiol (EU) was prepared by reacting urushiol with epichlorohydrin. Secondly, the EU was reacted with acrylic acid to obtain acrylic epoxide urushiol (AEU). Next, the prepolymers were synthesized by the reaction of AEU, 2,2-Bis(hydroxymethyl)propionic acid (DMPA), and isophorone diisocyanate (IPDI) and hexamethylene diisocyanate (HDI), respectively, using acetone as a solvent. The prepolymers were further neutralized by triethylamine (TEA) to obtain ionomers and dispersed in the water. After removing the acetone by vacuum distillation, the polyurethane dispersions (PUDs) were obtained. Finally, the WUV coatings were performed by adding a photoinitiator (Irgacure 2959). The products in the synthesized processes and the properties of the WUV coatings were examined. The results showed that the EU, AEU, prepolymers, and ionomers could be synthesized stably. The PUDs synthesized by the IPDI and HDI had a similar solid content of 25.2% and 26.2%, and similar pH values of 7.8 and 7.6. However, the IPDI-containing PUD displayed lower viscosity, smaller particle size, and a more even polydispersity index. The IPDI-containing WUV film displayed a higher hardness, gloss, and lightfastness. The HDI-containing WUV film possessed superior impact resistance. Both IPDI-containing and HDI-containing WUV films showed excellent adhesion, bending resistance, and mass retention, and demonstrated a potential for wood finishing.

In this study, a renewable polymeric material, refined oriental lacquer (ROL), used as a wood protective coating, and the Acacia confusa Merr. heartwood extractive, which was added as a natural photostabilizer for improving the lightfastness of ROL, were investigated. The best extract conditions for preparing heartwood extractives and the most suitable amount of addition (0, 1, 3, 5, and 10 phr) were investigated. The lightfastness index including brightness difference (ΔL *), yellowness difference (ΔYI), and color difference (ΔE *), and their applied properties of coating and film were measured. In the manufacture of heartwood extractives, the yield of extractives with acetone solvent was 9.2%, which was higher than that from toluene/ethanol solvent of 2.6%, and also had the most abundant total phenolic contents (535.2 mgGAE/g) and total flavonoid contents (252.3 μgRE/g). According to the SEM inspection and FTIR analysis, the plant gums migration to the surface of films and cracks occurred after UV exposure. The phenomena for photodegradation of ROL films were reduced after the addition of heartwood extractives. Among the different amounts of the heartwood extractives, the 10 phr addition was the best choice; however, the 1 phr heartwood extractive addition already showed noticeable lightfastness improvement. The drying times of ROL were extended and film performances worse with higher additions of heartwood extractives. Among the ROL films with different heartwood extractive additions, the ROL film with 1 phr addition had superior films properties, regarding adhesion and thermal stability, compared with the films of raw oriental lacquer.

Abstract Background Atypical subtrochanteric fracture and femoral fracture have been considered to be rare complications related to long-term bisphosphonates use. A reduced bone turnover rate may lead to delayed bone healing. Limited data have revealed that teriparatide treatment may reverse the effect of bisphosphonates and be effective in bone healing. Methods We reviewed patients with atypical subtrochanteric and femoral fracture related to bisphosphonates use between January 2008 and December 2014. Thirteen female patients were enrolled. Radiographic findings were compatible with the characteristics of atypical fracture. Surgical intervention was performed for all, and teriparatide use was advised postoperatively. Outcome measures included perioperative results, and clinical and radiographic outcome. Results Of the 13 female patients enrolled, 10 had subtrochanteric and 6 had proximal femoral fracture; 3 had bilateral fractures. The mean age of the patients at surgery was 70.15±6.36 years. Most fractures (68.8%) presented prodromal thigh pain. All patients were treated with an intramedullary fixation system without severe complications. The patients were divided into 2 groups based on whether they had received treatment with teriparatide or not. The mean time to bone union was 4.4 months in the teriparatide-treated group, and 6.2 months in the non-teriparatide-treated group (p=0.116). Six patients (75%) in the teriparatide-treated group and 4 (50%) in the non-teriparatide-treated group (p= 0.3) achieved bone union within 6 months. The means of the modified Harris Hip Score and Numerical Rating Scale were significantly better in the teriparatide-treated group at postoperative 6 months. Seven patients had the same ability to walk at the 1-year follow-up as they did before the atypical fracture. Conclusions Teriparatide treatment in patients with atypical fracture may help in fracture healing, hip function recovery, and pain relief in this reduced bone turnover patient group.

Summary: Pluripotent stem cells (PSCs) deficient for microRNAs (miRNAs), such as Dgcr8−/− or Dicer−/– embryonic stem cells (ESCs), contain no mature miRNA and cannot differentiate into somatic cells. How miRNA deficiency causes differentiation defects remains poorly understood. Here, we report that miR-302 is sufficient to enable neural differentiation of differentiation-incompetent Dgcr8−/− ESCs. Our data showed that miR-302 directly suppresses the tumor suppressor p53, which is modestly upregulated in Dgcr8−/− ESCs and serves as a barrier restricting neural differentiation. We demonstrated that direct inactivation of p53 by SV40 large T antigen, a short hairpin RNA against Trp53, or genetic ablation of Trp53 in Dgcr8−/− PSCs enables neural differentiation, while activation of p53 by the MDM2 inhibitor nutlin-3a in wild-type ESCs inhibits neural differentiation. Together, we demonstrate that a major function of miRNAs in neural differentiation is suppression of p53 and that modest activation of p53 blocks neural differentiation of miRNA-deficient PSCs. : In this article, Zhao and colleagues show that expression of miR-302 is sufficient to enable neural differentiation of differentiation-incompetent Dgcr8−/− ESCs. The authors demonstrated that miR-302 directly suppresses p53, which serves as a barrier restricting neural differentiation. Keywords: pluripotent stem cells, microRNA, differentiation, miR-302, p53, Dgcr8, neural differentiation, nutlin-3a, apoptosis

In our previous report, the antibacterial agents with different metals, mono(hydroxyethoxyethyl)phthalate [M(HEEP)2, M = Zn, Mn, and Ca], were synthesized. For increasing their yields, modified synthesis and purified processes were further investigated. The result of energy-dispersive X-ray spectroscopy showed the M(HEEP)2 could be stable and successfully synthesized, and their yields were raised to 73–85% from our previous report of 43–55%. For ultraviolet-cured wood floor coating application, the Zn(HEEP)2 was selected as an antibacterial agent and mixed with commercial UV wood floor coating. The effects on the antibacterial activity of UV films with different Zn(HEEP)2 additions of 0, 4, 8, and 12 phr as well as the commercial nano-Ag of 12 phr against Escherichia coli were evaluated. In the static antibacterial test, the UV films with Zn(HEEP)2 additions had similar antibacterial activity of 57–59%. In another dynamic shaking antibacterial test, the film containing 12 phr Zn(HEEP)2 had the best antibacterial activity among all the UV films. On the film properties, the Zn(HEEP)2-containing UV films had lower gloss and abrasion resistance, and slightly increased the hardness than those of UV film without Zn(HEEP)2 addition. However, there were no noticeable differences in mass retention, lightfastness, and thermal stability between UV films with and without the Zn(HEEP)2 addition. In this study, the 12 phr Zn(HEEP)2-containing UV film provided the best antibacterial activity against E. coli and had the balanced film properties for application on the UV wood floor coating.

High entropy oxides (HEOx) are novel materials, which increase the potential application in the fields of energy and catalysis. However, a series of HEOx is too novel to evaluate the synthesis properties, including formation and fundamental properties. Combining first-principles calculations with machine learning (ML) techniques, we predict the lattice constants and formation energies of spinel-structured photocatalytic HEOx, (Co,Cr,Fe,Mn,Ni)3O4, for stoichiometric and non-stoichiometric structures. The effects of site occupation by different metal cations in the spinel structure are obtained through first-principles calculations and ML predictions. Our predicted results show that the lattice constants of these spinel-structured oxides are composition-dependent and that the formation energies of those oxides containing Cr atoms are low. The computing time and computing energy can be greatly economized through the tandem approach of first-principles calculations and ML.

Polypropylene has been widely used as dielectric material in organic thin-film capacitors due to their high breakdown strength, low dielectric loss and self-healing capability. However, polypropylene’s energy density is relatively low. Increasing the energy density of polypropylene by adding materials with a high dielectric constant is commonly used. Still, it often leads to an increase in dielectric loss, lower dielectric strength and other shortcomings. In this study, a thin 2D platelet of mica/graphene oxide composite material was made from exfoliated mica as a substrate and attached by graphene oxide. The mica/graphene oxide platelets were added to polypropylene to make a plastic dielectric composite. The non-conductive flat inorganic additive can increase the dielectric constant and dielectric strength of the composite without increasing dielectric loss. The tiny mica/graphene oxide platelets can significantly improve the dielectric properties of polypropylene. The results show that by adding a small amount (less than 1 wt%) mica/graphene oxide, the relative dielectric constant of polypropylene can increase to more than 3.7 without causing an increase in dielectric loss and the dielectric strength of polypropylene can also enhance.

Oriental lacquer is a natural polymeric coating with a satiny texture and excellent characteristics, such as chemical resistance and durability. However, the poor lightfastness resulted in the natural aromatic structures of the urushiol structures limited its suitability for outdoor application. This study aimed at the improvement of the lightfastness by adding the different hindered amine light stafbilizers (HALS) with 2 phr addition as well as the effects on the coating and film property of the refined oriental lacquers (RL). The Cryptomeria japonica plate, glass sheets, and the other substrates were used for finishing. The results showed that the lightfastness of RL film was obviously improved by adding 2 phr HALS of Bis(2,2,6,6-tetramethyl-4-piperidinyl) sebacate (H90) containing -NH group and Bis (1,2,2,6,6-pentamethyl-4-piperidinyl)-[[3,5-bis(1,1-dimethyl ethyl) -4-hydroxyphenyl] methyl] butylmalonate (H60) containing -NCH3 groups. The HALS additions increased the pH value of RL and improved the activities of laccase-catalyzed reaction. Meanwhile, the oxidative polymerization of the side chains of RL was inhibited, caused by a radical scavenging ability of HALS. The changes in the drying process affected not only the curing time, but also the film properties. Among the 2 phr additions of different HALS, the film containing H90 had the best lightfastness. Meanwhile, it kept the most similar properties with RL and shortened the drying time of RL, and it was selected as the best HALS addition under 2 phr in this study.

Mutations of the Janus family kinase JAK3 gene cause severe combined immunodeficiency (SCID). JAK3 deficiency in humans is characterized by the absence of circulating T cells and natural killer (NK) cells with normal numbers of poorly functioning B cells (T–B+NK–). Using SCID patient-specific induced pluripotent stem cells (iPSCs) and a T cell in vitro differentiation system, we demonstrate a complete block in early T cell development of JAK3-deficient cells. Correction of the JAK3 mutation by CRISPR/Cas9-enhanced gene targeting restores normal T cell development, including the production of mature T cell populations with a broad T cell receptor (TCR) repertoire. Whole-genome sequencing of corrected cells demonstrates no CRISPR/Cas9 off-target modifications. These studies describe an approach for the study of human lymphopoiesis and provide a foundation for gene correction therapy in humans with immunodeficiencies.

Background: It remains unknown whether hearing loss increases the risk of Alzheimer’s disease. This study aimed to examine the association between hearing loss and risk of Alzheimer’s disease in older people in Taiwan. Methods: Analyzing the database from Taiwan’s National Health Insurance Program, this case-control study enrolled 488 subjects ≥65 years old with newly diagnosed Alzheimer’s disease as a case group and 1952 subjects without Alzheimer’s disease as a control group from 1998–2011. Patients with Alzheimer’s disease and other comorbidities were identified by analyzing ICD-9 coding in claims data. The association of hearing loss, other comorbidities, and risk of Alzheimer’s disease were compared between groups. Results: After controlling for confounders, multivariable logistic regression showed an adjusted odds ratio of Alzheimer’s disease of 1.39 in people with hearing loss (95% CI, 1.05–1.84) versus those without. Parkinson’s disease (OR 4.44; 95% CI, 2.54–7.78), head injury (OR 2.31; 95% CI, 1.46–3.66), depression (OR 1.68; 95% CI, 1.19–2.39), hypertension (OR 1.40; 95% CI, 1.10–1.79), and age (each year, OR 1.03; 95% CI, 1.01–1.05) also showed strong links with Alzheimer’s. Conclusions: Hearing loss is associated with increased risk of Alzheimer’s disease in older people in Taiwan.

The PI3K enhancer PIKE links PI3K catalytic subunits to group 1 metabotropic glutamate receptors (mGlu1/5) and activates PI3K signaling. The roles of PIKE in synaptic plasticity and the etiology of mental disorders are unknown. Here, we show that increased PIKE expression is a key mediator of impaired mGlu1/5-dependent neuronal plasticity in mouse and fly models of the inherited intellectual disability fragile X syndrome (FXS). Normalizing elevated PIKE protein levels in FXS mice reversed deficits in molecular and cellular plasticity and improved behavior. Notably, PIKE reduction rescued PI3K-dependent and -independent neuronal defects in FXS. We further show that PI3K signaling is increased in a fly model of FXS and that genetic reduction of the Drosophila ortholog of PIKE, CenG1A rescued excessive PI3K signaling, mushroom body defects, and impaired short-term memory in these flies. Our results demonstrate a crucial role of increased PIKE expression in exaggerated mGlu1/5 signaling causing neuronal defects in FXS.

Experience and activity refine cortical circuits through synapse elimination, but little is known about the activity patterns and downstream molecular mechanisms that mediate this process. We used optogenetics to drive individual mouse CA1 hippocampal neurons to fire in theta frequency bursts to understand how cell autonomous, postsynaptic activity leads to synapse elimination. Brief (1 hr) periods of postsynaptic bursting selectively depressed AMPA receptor (R) synaptic transmission, or silenced excitatory synapses, whereas more prolonged (24 hr) firing depressed both AMPAR and NMDAR EPSCs and eliminated spines, indicative of a synapse elimination. Both synapse silencing and elimination required de novo transcription, but only silencing required the activity-dependent transcription factors MEF2A/D. Burst firing induced MEF2A/D-dependent induction of the target gene Arc which contributed to synapse silencing and elimination. This work reveals new and distinct forms of activity and transcription-dependent synapse depression and suggests that these processes can occur independently.

In Taiwan, it is important to maintain sustainable development of the forestry industry in order to raise the self-sufficiency of domestic timber. Japanese cedar (Cryptomeria D. Don and Formosa acacia (Acacia confusa Merr.(Leguminosae)) have abundant storage options and are the potential candidates for this purpose. Heat treatment is a new environment-friendly method used to enhance the dimensional stability and durability of wood. On treatment, a surface with new characteristics is produced because of wood component changes. Consequently, an inactivated surface and a weak boundary layer are generated, and the wettability for adhesives and coatings is reduced. Furthermore, it decreases the pH value of the wood surface, and results in delay or acceleration during the curing of adhesives. This phenomenon must be paid attention to for practical applications of heat-treated wood. Ideal heat-treated conditions of C. japonica and A. confusa woods with productive parameters such as temperature, holding time, heating rate, and thicknesses of wood were identified in our previous study. In this research work, we focus on the normal shear strength of heat-treated wood with adhesives such as urea-formaldehyde resin (UF) and polyvinyl acetate (PVAc), and the finishing performances of heat-treated wood with polyurethane (PU) and nitrocellulose lacquer (NC) coatings as well as assessing the decay-resistance of heat-treated wood. The results show that heat-treated wood had a better decay resistance than untreated wood. The mass decrease of heat-treated wood was only 1/3 or even less than the untreated wood. The normal shear strength of heat-treated wood with UF and PVAc decreased from 99% to 72% compared to the untreated wood, but the wood failure of heat-treated wood was higher than that of the untreated one. Furthermore, the adhesion and impact resistance of wood finished by PU and NC coatings showed no difference between the heat-treated wood and untreated wood. The finished heat-treated wood had a superior durability and better gloss retention and lightfastness than that of the untreated wood.

The linseed oil glyceride (LOG) was synthesized by using a transesterification process with a glycerol/linseed oil molar ratio of 1.0. The waterborne urethane oil (WUO) wood coating was prepared by acetone process. First, dimethylolpropionic acid was reacted with hexamethylene diisocyanate (HDI) or isophorone diisocyanate (IPDI), followed by adding LOG at various NCO/OH molars of 0.7, 0.8, and 0.9, respectively, and the COOH-containing prepolymer was obtained. Then, the ionomer which was prepared by neutralizing prepolymer with trimethylamine, was dispersed by adding deionized water, and the water⁻acetone dispersion was obtained. Finally, the acetone was removed by vacuum distillation. In the whole synthesized process, the LOG and COOH-containing prepolymer could be steadily synthesized by FTIR analysis, and the weight-average molecular weight and polydispersity of COOH-containing prepolymer increased with an increase of NCO/OH molar ratios. During the water dispersion process of the ionomer acetone solution, the point of phase inversion was prolonged, meaning the solid content decreased with an increase of NCO/OH molar ratios. After acetone was removed, the color of WUO was milky-white, and it was weakly alkaline and possessed a pseudoplastic fluid behavior. The particle size of WUO increased with increasing of NCO/OH molar ratios, however, the storage stability was extended for HDI and shortened for IPDI synthesized with increasing of NCO/OH molar ratios.

In this study, the distributed consensus control and model predictive control (MPC)-based formation strategies for quadrotors are proposed. First, the formation-control problem is decoupled into horizontal and vertical motions. The distributed consensus control and MPC-based formation strategy are implemented in the follower’s horizontal formation control. In the horizontal motion, the leader tracks the given waypoints by simply using the MPC, and generates the desired formation trajectory for each follower based on its flight information, predicted trajectory, and the given formation pattern. On the other hand, the followers carry out the formation flight based on the proposed horizontal formation strategy and the desired formation trajectories generated by the leader. In the vertical motion, formation control is carried out using only the MPC for both the leader and the follower. Likewise, the leader tracks the desired altitude/climb rate and generates the desired formation trajectories for the followers, and the followers track the desired formation trajectories generated by the leader using the MPC. The optimization problem considered in the MPC differs for the horizontal and vertical motions. The problem is formulated as a quadratic programming (QP) problem for the horizontal motion, and as a linear quadratic tracker (LQT) for the vertical motion. Simulation of a comprehensive maneuver was carried out under a Matlab/Simulink environment to examine the performance of the proposed formation strategies.

In this work, silver nanoparticles (Ag NPs) were decorated on thiol (–SH) grafted graphene oxide (GO) layers to investigate the antibacterial activities in Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (Pseudomonas aeruginosa). The quasi-spherical, nano-sized Ag NPs were attached to the GO surface layers, as confirmed by using field emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM), respectively. The average size of GO-Ag nanocomposites was significantly reduced (327 nm) from those of pristine GO (962 nm) while the average size of loaded Ag NPs was significantly smaller than the Ag NPs without GO. Various concentrations of AgNO3 solutions (0.1, 0.2, and 0.25 M) were loaded into GO nanosheets and resulted in the Ag contents of 31, 43, and 65%, respectively, with 1–2 nm sizes of Ag NPs anchored on the GO layers. These GO-Ag samples have negative surface charges but the GO-Ag 0.2 M sample (43% Ag) demonstrated the highest antibacterial efficiency. At 10 ppm load of GO-Ag suspension, only a GO-Ag 0.2 M sample yielded slight bacterial inhibition (5.79–7.82%). As the GO-Ag content was doubled to 20 ppm, the GO-Ag 0.2 M composite exhibited ~49% inhibition. When the GO-Ag 0.2 M composite level was raised to 100 ppm, almost 100% inhibition efficiencies were found on both Staphylococcus aureus (S.A.) and Pseudomonas aeruginosa (P.A.), which were significantly higher than using pristine GO (27% and 33% for S.A. and P.A.). The combined effect of GO and Ag nanoparticles demonstrate efficient antibacterial activities.

Human induced pluripotent stem cells (hiPSCs) have enormous potential for the treatment of inherited and acquired disorders. Recently, antigen-specific T lymphocytes derived from hiPSCs have been reported. However, T lymphocyte populations with broad T cell receptor (TCR) diversity have not been generated. We report that hiPSCs derived from skin biopsy are capable of producing T lymphocyte populations with a broad TCR repertoire. In vitro T cell differentiation follows a similar developmental program as observed in vivo, indicated by sequential expression of CD7, intracellular CD3 and surface CD3. The γδ TCR locus is rearranged first and is followed by rearrangement of the αβ locus. Both γδ and αβ T cells display a diverse TCR repertoire. Upon activation, the cells express CD25, CD69, cytokines (TNF-α, IFN-γ, IL-2) and cytolytic proteins (Perforin and Granzyme-B). These results suggest that most, if not all, mechanisms required to generate functional T cells with a broad TCR repertoire are intact in our in vitro differentiation protocol. These data provide a foundation for production of patient-specific T cells for the treatment of acquired or inherited immune disorders and for cancer immunotherapy.

This study investigated the functional roles of the N-terminal Ca(2+) ion-binding sites, in terms of enzyme catalysis and stability, of peptidylarginine deiminase 4 (PAD4). Amino acid residues located in the N-terminal Ca(2+)-binding site of PAD4 were mutated to disrupt the binding of Ca(2+) ions. Kinetic data suggest that Asp155, Asp157 and Asp179, which directly coordinate Ca3 and Ca4, are essential for catalysis in PAD4. For D155A, D157A and D179A, the k(cat)/K(m,BAEE) values were 0.02, 0.63 and 0.01 s(-1)mM(-1) (20.8 s(-1)mM(-1) for WT), respectively. Asn153 and Asp176 are directly coordinated with Ca3 and indirectly coordinated with Ca5 via a water molecule. However, N153A displayed low enzymatic activity with a k(cat) value of 0.3 s(-1) (13.3 s(-1) for wild-type), whereas D176A retained some catalytic power with a k(cat) of 9.7 s(-1). Asp168 is the direct ligand for Ca5, and Ca5 coordination by Glu252 is mediated by two water molecules. However, mutation of these two residues to Ala did not cause a reduction in the k(cat)/K(m,BAEE) values, which indicates that the binding of Ca5 may not be required for PAD4 enzymatic activity. The possible conformational changes of these PAD4 mutants were examined. Thermal stability analysis of the PAD4 mutants in the absence or presence of Ca(2+) indicated that the conformational stability of the enzyme is highly dependent on Ca(2+) ions. In addition, the results of urea-induced denaturation for the N153, D155, D157 and D179 series mutants further suggest that the binding of Ca(2+) ions in the N-terminal Ca(2+)-binding site stabilizes the overall conformational stability of PAD4. Therefore, our data strongly suggest that the N-terminal Ca(2+) ions play critical roles in the full activation of the PAD4 enzyme.

DNA replication in mammals is regulated via the coordinate firing of clusters of replicons that duplicate megabase-sized chromosome segments at specific times during S-phase. Cytogenetic studies show that these "replicon clusters" coalesce as subchromosomal units that persist through multiple cell generations, but the molecular boundaries of such units have remained elusive. Moreover, the extent to which changes in replication timing occur during differentiation and their relationship to transcription changes has not been rigorously investigated. We have constructed high-resolution replication-timing profiles in mouse embryonic stem cells (mESCs) before and after differentiation to neural precursor cells. We demonstrate that chromosomes can be segmented into multimegabase domains of coordinate replication, which we call "replication domains," separated by transition regions whose replication kinetics are consistent with large originless segments. The molecular boundaries of replication domains are remarkably well conserved between distantly related ESC lines and induced pluripotent stem cells. Unexpectedly, ESC differentiation was accompanied by the consolidation of smaller differentially replicating domains into larger coordinately replicated units whose replication time was more aligned to isochore GC content and the density of LINE-1 transposable elements, but not gene density. Replication-timing changes were coordinated with transcription changes for weak promoters more than strong promoters, and were accompanied by rearrangements in subnuclear position. We conclude that replication profiles are cell-type specific, and changes in these profiles reveal chromosome segments that undergo large changes in organization during differentiation. Moreover, smaller replication domains and a higher density of timing transition regions that interrupt isochore replication timing define a novel characteristic of the pluripotent state.

This study aimed to develop e-learning materials for a blended learning course focusing on programmable logic controller (PLC) technology and assessed its effectiveness using the technology acceptance model. The course utilized the university's cloud-based learning platform as the teaching platform and incorporated specially designed targeting PLC technology. The objective was to enhance the interactivity and efficiency of the instructional activities while fostering the students' interest in programmable control technology. This study placed emphasis on helping students establish a solid foundation in programmable control technology and improving learning outcomes, thereby achieving the goals of blended instructional design. The results of questionnaire survey were analyzed using statistical methods. Structural equation modeling using Smart PLS software was used to measure the structural model. A number ofconclusions were drawn based on the analysis results. First, the developed e-learning material effectively enhanced the learners' interest and learning outcomes. Second. the students generally exhibited high levels of satisfaction with the blended learning course in PLC technology. indicating widespread recognition and affirmation of the course. Third, the blended course for PLC technology demonstrated applicability in the fields of engineering and technology education. [ABSTRACT FROM AUTHOR]