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OBJECTIVE: To explore the clinical characteristics and variant of CREBBP gene in a fetus with Rubinstein-Taybi syndrome (RSTS). METHODS: A fetus with RSTS diagnosed at the Third Affiliated Hospital of Zhengzhou University in August 2022 was selected as the study subject. Clinical data, amniotic fluid sample of the fetus and peripheral blood samples of its parents were collected for whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. RESULTS: Foot malformation, cerebellar vermis agenesis, brain agenesis, polysyndactyly of the big toes and other phenotypes were found by prenatal ultrasound. WES revealed that the fetus has harbored a heterozygous c.4684G>T (p.E1562*) variant in exon 28 of the CREBBP gene (NM_004380.3), which was de novo in origin. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be pathogenic (PVS1+PS2_Moderate+PM2_Supporting). After genetic counseling, the couple had opted to terminate the pregnancy and refused autopsy of the fetus. CONCLUSION: The c.4684G>T (p.E1562*) variant of the CREBBP gene probably underlay the RSTS in this fetus. The newly discovered variant has enriched the mutational spectrum of the CREBBP gene and illustrated that WES is an efficient tool for the prenatal diagnosis of RSTS.
Subject(s)
CREB-Binding Protein , Exome Sequencing , Prenatal Diagnosis , Rubinstein-Taybi Syndrome , Humans , Rubinstein-Taybi Syndrome/genetics , Female , Pregnancy , CREB-Binding Protein/genetics , Adult , Fetus/abnormalities , Fetus/diagnostic imaging , Mutation , Male , Ultrasonography, PrenatalABSTRACT
Background: This cohort study was conducted to devise an approximation formula for predicting the glomerular filtration rate (GFR) after 1 year using annual medical examination data from the general population. Methods: Consecutive annual medical examination data were obtained for 41,337 inhabitants. Machine learning with the random forest method was used to assess the importance of each clinical parameter in terms of its association with estimated GFR (eGFR) after 1 year. An approximation formula was developed by multiple linear regression analysis based on the four most important clinical parameters. The relationship between the GFR after 1 year approximated by our formula and the eGFR after 1 year was analyzed using Pearson's correlation coefficient. Results: The following approximation formula was obtained by multiple linear regression analysis: approximate GFR after 1 year (mL/min/1.73 m2) = -0.054 × age + 0.162 × hemoglobin - 0.085 × uric acid + 0.849 × eGFR + 11.5. The approximate GFR after 1 year was significantly and strongly correlated with the eGFR at that time (r = 0.884; p < 0.001). Conclusions: An approximation formula including age, hemoglobin, uric acid, and eGFR may be useful for predicting GFR after 1 year among members of the general population.
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Person-centered primary care measures (PCPCM) facilitate high-quality and culturally appropriate primary care. Access to PCPCM remains unequal between rural and urban areas, and the available evidence on rural PCPCM is still lacking. A cross-sectional survey was conducted with stratified sampling by regions, and four districts (Xicheng, Fengtai, Huairou, and Daxing) in Beijing were selected to test the performance of PCPCM in both urban and rural areas. Descriptive statistical methods were used to compare the urban-rural differences in the demographic characteristics of PCPCM. Correlation and regression analyses were performed to determine the associations between PCPCM in demographics and utilization of primary care. The PCPCM showed good reliability and validity in both urban and rural areas (P < .001), slightly lower in rural areas, but scores of rural PCPCM (R-PCPCM) in all items were lower than urban PCPCM (U-PCPCM). Patients in either the preferred urban or rural health centers all showed the highest PCPCM scores, with U-PCPCM= 3.31 for CHCs and R-PCPCM= 3.10 for RHCs, respectively. Patients in urban areas were more likely to receive higher-quality primary care than in rural areas (P < .001). Patients who preferred hospitals (ß = 2.61, P < .001) or CHCs (ß = 0.71, P = .003) as providers was a significant positive predictor of U-PCPCM but it was the preference for hospitals (ß = 2.95, P < .001) for R-PCPCM. Urban-rural differences existed in the performance of PCPCM, with rural areas typically more difficult to access better PCPCM. To promote health equity in rural areas, healthcare providers should strive to minimize urban-rural differences in the quality and utilization of primary care services as much as feasible.
Subject(s)
Health Equity , Primary Health Care , Rural Population , Humans , Primary Health Care/statistics & numerical data , Cross-Sectional Studies , Male , Female , Middle Aged , Adult , Rural Population/statistics & numerical data , Beijing , Patient-Centered Care/statistics & numerical data , Urban Population/statistics & numerical data , Health Services Accessibility/statistics & numerical data , China , Aged , Quality of Health Care , Rural Health Services/statistics & numerical data , Surveys and Questionnaires , Reproducibility of ResultsABSTRACT
BACKGROUND: Dendritic cells (DCs) regulate the immune response associated with T lymphocytes, but their role in stroke remains unclear. In this study, we investigated the causal relationship between DCs and T-cell response in intracerebral hemorrhage (ICH) by focusing on TLRs (toll-like receptors) that may modulate the function of DCs. METHODS: We studied the effects of TLR4, TLR2, and TLR9 on DC-mediated T-cell response and the outcomes of ICH using male C57BL/6 and CD11c-DTx (diphtheria toxin) receptor mice. We administered specific agents intraperitoneally or orally and evaluated the results using flow cytometry, real-time polymerase chain reaction, Western blotting, immunofluorescence staining, histopathology, and behavioral tests. RESULTS: TLR4 and TLR2 activation induces DC maturation and reduces the ratio of regulatory T to T-helper 17 cells in the brain and periphery after ICH. When either of these receptors is activated, it can worsen neuroinflammation and exacerbate ICH outcomes. TLR9 also promotes DC maturation, stabilizing the number of DCs, particularly conventional DCs. TLR9 has the opposite effects on regulatory T/T-helper 17 balance, neuroinflammation, and ICH outcomes compared with TLR4 and TLR2. Upon stimulation, TLR4 and TLR9 may achieve these effects through the p38-MAPK (p38-mitogen-activated protein kinase)/MyD88 (myeloid differentiation primary response gene 88) and indoleamine 2,3-dioxygenase 1 (IDO1)/GCN2 (general control nonderepressible 2) signaling pathways, respectively. DCs act as intermediaries for TLR-mediated T-cell response. CONCLUSIONS: TLR-mediated opposing effects of DCs on T-cell response may provide novel strategies to treat ICH.
Subject(s)
Cerebral Hemorrhage , Dendritic Cells , Mice, Inbred C57BL , T-Lymphocytes, Regulatory , Th17 Cells , Animals , Cerebral Hemorrhage/immunology , Cerebral Hemorrhage/metabolism , Dendritic Cells/immunology , T-Lymphocytes, Regulatory/immunology , Mice , Th17 Cells/immunology , Male , Toll-Like Receptors/metabolism , Toll-Like Receptor 2/metabolism , Toll-Like Receptor 9/metabolism , Toll-Like Receptor 4/metabolism , Toll-Like Receptor 4/immunologyABSTRACT
BACKGROUND: With the conflict between the promise of ageing in health and longevity and the limited availability of health resources and social support, older adults in China inevitably experience anxieties surrounding health risks. This study aims to investigate how older adults perceive the health risks that come with getting older, explore the degree to which health risks affect older adults, and advocate for active engagement in practices for managing health risks. METHODS: Using purposive sampling, three districts of Beijing (Xicheng District, Fengtai District, and Daxing District, respectively) were selected for the research. Qualitative semi-structured and in-depth interviews were conducted with 70 community-dwelling older adults who participated in the study. Data were extracted and analyzed based on a thematic framework approach. RESULTS: Three main themes were identified: (i) the anxieties of older adults concerning health risks in ageing; (ii) the priorities of older adults for health risk management in ageing; (iii) the expectations of older adults for health risk management in ageing. The primary health concerns among older adults included disease incidence and function decline. It was found that basic health management emerged as a critical need for older adults to mitigate health risks. Moreover, it was observed that healthcare support for older adults from familial, institutional, and governmental levels exhibited varying degrees of inadequacy. CONCLUSIONS: The primary source of anxieties among older adults regarding health risks predominantly stems from a perceived sense of health deprivation. It is often compounded by persistent barriers to primary care of priorities in managing health risks among older adults. In addition, the expectations of older adults for health risk management emphasize the necessity for integrated care approaches. Therefore, further research should give priority to the prevention and management of health risks, aim to reduce anxieties, provide integrated care to meet the primary needs and expectations of older adults, and ultimately strive toward the overarching goal of promoting health and longevity.
Subject(s)
Anxiety , Independent Living , Qualitative Research , Humans , Aged , Female , Male , Independent Living/psychology , Aged, 80 and over , Anxiety/psychology , Anxiety/epidemiology , Middle Aged , Aging/psychology , Interviews as Topic , China/epidemiology , Risk Assessment , Health PrioritiesABSTRACT
BACKGROUND: Bawei Chenxiang Wan (BCW) is among the most effective and widely used therapies for coronary heart disease and angina pectoris in Tibet. However, whether it confers protection through a right-ventricle (RV) myocardial metabolic mechanism is unknown. METHODS: Male Sprague-Dawley rats were orally administrated with BCW, which was injected concurrently with a bolus of Sugen5416, and subjected to hypoxia exposure (SuHx; 5000 m altitude) for 4 weeks. Right ventricular hypertrophy (RVH) in high-altitude heart disease (HAHD) was assessed using Fulton's index (FI; ratio of RV to left ventricle + septum weights) and heart-weight-to-body-weight ratio (HW/BW). The effect of therapeutic administration of BCW on the RVH hemodynamics was assessed through catheterization (mean right ventricular pressure and mean pulmonary artery pressure (mRVP and mPAP, respectively)). Tissue samples were used to perform histological staining, and confirmatory analyses of mRNA and protein levels were conducted to detect alterations in the mechanisms of RVH in HAHD. The protective mechanism of BCW was further verified via cell culture. RESULTS: BCW considerably reduced SuHx-associated RVH, as indicated by macro morphology, HW/BW ratio, FI, mPAP, mRVP, hypertrophy markers, heart function, pathological structure, and myocardial enzymes. Moreover, BCW can alleviate the disorder of glucose and fatty acid metabolism through upregulation of carnitine palmitoyltransferase1É, citrate synthase, and acetyl-CoA and downregulation of glucose transport-4, phosphofructokinase, and pyruvate, which resulted in the reduced levels of free fatty acid and lactic acid and increased aerobic oxidation. This process may be mediated via the regulation of sirtuin 3 (SIRT3)-hypoxia-inducible factor 1α (HIF1α)-pyruvate dehydrogenase kinase (PDK)/pyruvate dehydrogenase (PDH) signaling pathway. Subsequently, the inhibition of SIRT3 expression by 3-TYP (a selective inhibitor of SIRT3) can reverse substantially the anti-RVH effect of BCW in HAHD, as indicated by hypertrophy marker and serum myocardial enzyme levels. CONCLUSIONS: BCW prevented SuHx-induced RVH in HAHD via the SIRT3-HIF1É-PDK/PDH signaling pathway to alleviate the disturbance in fatty acid and glucose metabolism. Therefore, BCW can be used as an alternative drug for the treatment of RVH in HAHD.
Subject(s)
Drugs, Chinese Herbal , Hypertrophy, Right Ventricular , Hypoxia-Inducible Factor 1, alpha Subunit , Rats, Sprague-Dawley , Animals , Rats , Altitude Sickness/complications , Altitude Sickness/drug therapy , Disease Models, Animal , Drugs, Chinese Herbal/pharmacology , Fatty Acids/metabolism , Glucose/metabolism , Hypertrophy, Right Ventricular/drug therapy , Hypoxia-Inducible Factor 1, alpha Subunit/drug effects , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Pyruvate Dehydrogenase Acetyl-Transferring Kinase/drug effects , Signal Transduction/drug effects , Sirtuin 3/drug effects , Sirtuin 3/metabolismABSTRACT
Epidural fluid collection is commonly seen as a low-density accumulation beneath the dura on postoperative computed tomography scans. It is discussed less in the literature because, in most cases, the fluid amount is small, self-absorbing, and typically does not cause neurological deficits, so intervention is usually unnecessary. However, when a significant amount of fluid accumulates, patients may experience symptoms such as altered consciousness and even coma. In such cases, treatment is necessary to reduce intracranial pressure, preventing further deterioration of consciousness and potentially life-threatening situations. The authors report a case of a large epidural fluid collection following cranioplasty, resulting in progressive deterioration of consciousness in the patient. Computed tomography scans indicated brain herniation. Subsequently, percutaneous puncture and suction treatment were performed, followed by appropriate pressure dressing. The patient gradually recovered from a shallow coma to clear consciousness and was discharged after rehabilitation.
ABSTRACT
This study observed the effects of Notoginseng Radix et Rhizoma on the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin complex 1(mTORC1) signaling pathway and mitochondrial energy metabolism in the rat model of adriamycin-induced renal fibrosis with blood stasis syndrome to explore the mechanism of Notoginseng Radix et Rhizoma in protecting the kidney. Thirty male rats with adriamycin-induced renal fibrosis were randomized into model, low-, medium-, and high-dose Notoginseng Radix et Rhizoma, and positive control groups(n=6). Six clean SD male rats were selected into the normal group. The normal group and model group were administrated with normal saline, and other groups with corresponding drugs. After 8 weeks of treatment, the renal function, renal pathology, adenosine triphosphate(ATP) levels, Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)-ATPase activities, and the protein levels of ATP5B, mTORC1, 70 kDa ribosomal protein S6 kinase(P70S6K), P85, Akt, p-Akt, and SH2-containing inositol phosphatase(SHIP2) in the renal tissue were determined. Compared with the normal group, the model group showed elevated levels of blood urea nitrogen(BUN) and serum creatinine(SCr)(P<0.01). Compared with the model group, Notoginseng Radix et Rhizoma and the positive control lowered the levels of BUN and SCr, which were significant in the medium-and high-dose Noto-ginseng Radix et Rhizoma groups and the positive control group(P<0.05). Compared with the model group, Notoginseng Radix et Rhizoma and the positive control alleviated the pathological changes in the renal tissue, such as vacuolar and fibroid changes, glomerulus atrophy, cystic expansion of renal tubules, and massive infiltration of inflammatory cells. Compared with the normal group, the model group showed decreased mitochondrial ATP content and Na~+-K~+-ATPase and Ca~(2+)-Mg~(2+)-ATPase activities in the renal tissue(P<0.05), and medium-and high-dose Notoginseng Radix et Rhizoma and positive control mitigated such decreases(P<0.05). Compared with the model group, medium-and high-dose Notoginseng Radix et Rhizoma and the positive control up-regulated the protein levels of ATP5B and SHIP2 and down-regulated the protein levels of mTORC1, P70S6K, P85, Akt, and p-Akt(P<0.05 or P<0.01 or P<0.001). Notoginseng Radix et Rhizoma may exert an anti-fibrosis effect by inhibiting the activation of the PI3K/Akt/mTORC1 pathway to restore mitochondrial energy metabolism, thus protecting the kidney.
Subject(s)
Drugs, Chinese Herbal , Energy Metabolism , Mechanistic Target of Rapamycin Complex 1 , Mitochondria , Panax notoginseng , Proto-Oncogene Proteins c-akt , Rats, Sprague-Dawley , Animals , Male , Rats , Proto-Oncogene Proteins c-akt/metabolism , Proto-Oncogene Proteins c-akt/genetics , Mechanistic Target of Rapamycin Complex 1/metabolism , Panax notoginseng/chemistry , Mitochondria/drug effects , Mitochondria/metabolism , Energy Metabolism/drug effects , Drugs, Chinese Herbal/pharmacology , Drugs, Chinese Herbal/administration & dosage , Phosphatidylinositol 3-Kinases/metabolism , Phosphatidylinositol 3-Kinases/genetics , Rhizome/chemistry , Humans , Signal Transduction/drug effects , Kidney/drug effects , Kidney/metabolism , Renal Insufficiency/drug therapy , Renal Insufficiency/metabolismABSTRACT
Prime editing is a programmable genetic method that can precisely generate any desired small-scale variations in cells without requiring double-strand breaks and DNA donors. However, higher editing efficiency is greatly desirable for wide practical applications. In this study, we developed a target-specific prime editing reporter (tsPER) and a universal prime editing reporter (UPER) to facilitate rapid selection of desired edited cells through puromycin screening. The modification efficiency of HEK3_i1CTT_d5G in HEK293T cells improved from 36.37 % to 64.84 % with the incorporation of tsPER. The target sequence of interested genes could be custom inserted into a selection cassette in tsPER to establish personalized reporters. The UPER demonstrated PE3 editing efficiency up to 74.49 % on HEK3_i1CTT_d5G and 73.52 % on HEK3_i1His6, achieved through co-selection with an additional pegRNA (puro) to repair the mutant PuroR cassette. Overall, tsPER and UPER robustly improved the efficiency of prime editing. Both of these approaches expand enrichment strategies for genomically modified cells and accelerate the generation of genetically modified models.
Subject(s)
Gene Editing , Humans , Gene Editing/methods , HEK293 Cells , Genes, Reporter , CRISPR-Cas Systems , Puromycin/pharmacologyABSTRACT
Metal-organic frameworks (MOFs) are potential candidates for gas-selective adsorbents for the separation of an ethylene/ethane mixture. To accelerate material discovery, high-throughput computational screening is a viable solution. However, classical force fields, which were widely employed in recent studies of MOF adsorbents, have been criticized for their failure to cover complicated interactions such as those involving π electrons. Herein, we demonstrate that machine learning force fields (MLFFs) trained on quantum-chemical reference data can overcome this difficulty. We have constructed a MLFF to accurately predict the adsorption energies of ethylene and ethane on the organic linkers of MOFs and discovered that the π electrons from both the ethylene molecule and the aromatic rings in the linkers could substantially influence the selectivity for gas adsorption. Four kinds of MOF linkers are identified as having promise for the separation of ethylene and ethane, and our results could also offer a new perspective on the design of MOF building blocks for diverse applications.
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INTRODUCTION: Physical exercise (PE) is essential for alleviating the symptoms of sarcopenia. Low motivation is a major barrier to PE. Mindfulness-based intervention (MBI) has the potential to improve motivation. However, few studies have used a mindfulness-based PE (MBPE) intervention among older people with sarcopenia. OBJECTIVES: To assess the feasibility, acceptability and preliminary effects of the MBPE program among community-dwelling older people with sarcopenia. METHODS AND ANALYSIS: A two-arm pilot randomised controlled trial will be conducted to assess the feasibility, acceptability and preliminary effects of an MBPE program among community-dwelling older people with sarcopenia. A total of 60 participants will be randomised into the intervention group, receiving the MBPE intervention twice a week over 12 weeks, or the control group, receiving health education with the same duration, number of sessions and frequency as the intervention group. Each session of the MBPE program will last about 60 min, including 5-10- min introduction, 20-min MBI, 30-min PE and 5-10-min sharing and discussion. The primary outcomes will be the feasibility (i.e., the time spent recruiting participants, the eligibility rate and the recruitment rate) and acceptability (i.e., the attendance rate, completion rate and attrition rate) of the MBPE program. The secondary outcomes will be the preliminary effects of the MBPE program on symptoms of sarcopenia, motivation for PE, psychological well-being, mindfulness level, physical activity level and quality of life. Individual interviews will be conducted to identify the strengths, limitations and therapeutic components of the intervention. The quantitative data will be analysed by generalised estimating equations. The qualitative data will be analysed by Braun and Clarke's thematic approach. CONCLUSION: The findings of this study will be able to provide evidence for the health professionals in adopting MBPE as a supportive intervention for the older adults with sarcopenia and the groundworks for the researchers in developing non-pharmacological intervention for older adults. The positive effects could facilitate healthy ageing and relief the burden of the medical system, especially in the countries facing the ageing population. TRIAL REGISTRATION NUMBER: NCT05982067; ClinicalTrials.gov.
Subject(s)
Mindfulness , Sarcopenia , Humans , Aged , Sarcopenia/therapy , Quality of Life , Independent Living , Feasibility Studies , Pilot Projects , Exercise , Randomized Controlled Trials as TopicABSTRACT
Introduction: Cells expressing taste signaling elements in non-gustatory tissues have been described as solitary chemosensory cells (SCCs) or tuft cells. These "taste-like" cells play a critical role in the maintenance of tissue homeostasis. Although the expression of SCC markers and taste signaling constituents has been identified in mouse gingivae, their role in periodontal homeostasis is still unclear. Methods: Public RNA sequencing datasets were re-analyzed and further validated with RT-PCR/qRT-PCR and immunofluorescent staining to explore the expression of TAS2Rs and downstream signaling constituents in mouse gingival fibroblasts (MGFs). The specific action of salicin on MGFs via Tas2r143 was validated with RNA silence, heterologous expression of taste receptor/Gα-gustducin and calcium imaging. The anti-inflammatory effects of salicin against LPS-induced MGFs were investigated in cell cultures, and were further validated with a ligature-induced periodontitis mouse model using Ga-gustducin-null (Gnat3-/-) mice. Results: The expression of Tas2r143, Gnat3, Plcb2, and TrpM5 was detected in MGFs. Moreover, salicin could activate Tas2r143, elicited taste signaling and thus inhibited LPS-induced chemokines expression (CXCL1, CXCL2, and CXCL5) in MGFs. Consistently, salicin-treatment inhibited periodontal bone loss, inflammatory/chemotactic factors expression, and neutrophil infiltration in periodontitis mice, while these effects were abolished in Gnat3-/- mice. Discussion: Gingival fibroblasts play a critical role in the maintenance of periodontal homeostasis via "SCC-like" activity. Salicin can activate Tas2r143-mediated bitter taste signaling and thus alleviate periodontitis in mouse, indicating a promising approach to the resolution of periodontal inflammation via stimulating the "SCC-like" function of gingival fibroblasts.
Subject(s)
Benzyl Alcohols , Fibroblasts , Glucosides , Periodontitis , Transducin , Animals , Mice , Fibroblasts/metabolism , Lipopolysaccharides , Periodontitis/drug therapy , Periodontitis/metabolismABSTRACT
Breast cancer (BC) comprises multiple biological and histologic properties. MicroRNAs show key functions in cancer prognosis. This paper explored the relationship between miR-497-5p with clinicopathological characteristics and prognosis in BC. Cancer tissues and normal adjacent tissues (NATs) were collected from 140 included patients with BC. The clinical baseline data, including age, tumor size, pathologic grade, clinical stage, modified Scraff-Bloom-Richardson grade, and lymph node metastasis, were recorded. miR-497-5p expression in cancer tissues and NAT was determined by reverse transcription-quantitative polymerase chain reaction. Patients with BC were followed up for 5 years to record their survival. Patients were divided into the miR-497-5p low expression and high expression groups to assess the correlation between miR-497-5p expression with clinicopathological characteristics and overall survival of patients. The role of miR-497-5p as an independent risk factor for death was further analyzed by a multivariate Cox regression model. miR-497-5p was downregulated in BC tissues than NAT. Tumor size, clinical stage, and lymph node metastasis showed significant differences among patients with high and low miR-497-5p expression levels. Patients with BC with low miR-497-5p expression presented decreased survival. Lowly-expressed miR-497-5p was an independent risk factor for death in patients. Collectively, cancer tissue miR-497-5p low expression increases the risk of death and serves as an independent risk factor for death in patients with BC.
Subject(s)
Breast Neoplasms , MicroRNAs , Humans , Female , Breast Neoplasms/genetics , Breast Neoplasms/pathology , Lymphatic Metastasis , Biomarkers, Tumor , Gene Expression Regulation, Neoplastic , MicroRNAs/geneticsABSTRACT
A facile and efficient copper-catalyzed domino-double annulation strategy was developed from easily accessible o-aminobenzamides and 2-iodoisothiocyanates, which affords a direct pathway for the synthesis of tetracyclic fused 12H-benzo[4,5]thiazolo[2,3-b]quinazolin-12-ones in moderate to good yields without the addition of ligands, bases, and external oxidants. The reaction involves a C-N bond cleavage and the formation of a C-N/C-S bond in one step with the advantages of using an inexpensive copper catalyst and easy operation. Mechanistic studies suggest that this transformation proceeds via intermolecular condensation of o-aminobenzamides with 2-iodoisothiocyanates, followed by an intramolecular Ullmann-type cross-coupling cyclization reaction.
ABSTRACT
A palladium-catalyzed intermolecular [2 + 2 + 2] oxidative coupling-annulation of terminal alkenes and alkynes using copper(II) as the oxidant has been developed through direct C-C bond formation. These reactions provide effective access to multiaryl-substituted benzenes with high regioselectivity in the absence of any ligands. The features of this protocol are broad substrate scope, and high atom and step economy. The aggregation-induced emission properties of selected products were further investigated. These synthesized multiaryl-substituted benzenes may be worth exploring for further applications in the fields of advanced functional materials or drugs.
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As an intrinsic cellular mechanism responsible for the internalization of extracellular ligands and membrane components, caveolae-mediated endocytosis (CavME) is also exploited by certain pathogens for endocytic entry [e.g., Newcastle disease virus (NDV) of paramyxovirus]. However, the molecular mechanisms of NDV-induced CavME remain poorly understood. Herein, we demonstrate that sialic acid-containing gangliosides, rather than glycoproteins, were utilized by NDV as receptors to initiate the endocytic entry of NDV into HD11 cells. The binding of NDV to gangliosides induced the activation of a non-receptor tyrosine kinase, Src, leading to the phosphorylation of caveolin-1 (Cav1) and dynamin-2 (Dyn2), which contributed to the endocytic entry of NDV. Moreover, an inoculation of cells with NDV-induced actin cytoskeletal rearrangement through Src to facilitate NDV entry via endocytosis and direct fusion with the plasma membrane. Subsequently, unique members of the Rho GTPases family, RhoA and Cdc42, were activated by NDV in a Src-dependent manner. Further analyses revealed that RhoA and Cdc42 regulated the activities of specific effectors, cofilin and myosin regulatory light chain 2, responsible for actin cytoskeleton rearrangement, through diverse intracellular signaling cascades. Taken together, our results suggest that an inoculation of NDV-induced Src-mediated cellular activation by binding to ganglioside receptors. This process orchestrated NDV endocytic entry by modulating the activities of caveolae-associated Cav1 and Dyn2, as well as specific Rho GTPases and downstream effectors. IMPORTANCE: In general, it is known that the paramyxovirus gains access to host cells through direct penetration at the plasma membrane; however, emerging evidence suggests more complex entry mechanisms for paramyxoviruses. The endocytic entry of Newcastle disease virus (NDV), a representative member of the paramyxovirus family, into multiple types of cells has been recently reported. Herein, we demonstrate the binding of NDV to induce ganglioside-activated Src signaling, which is responsible for the endocytic entry of NDV through caveolae-mediated endocytosis. This process involved Src-dependent activation of the caveolae-associated Cav1 and Dyn2, as well as specific Rho GTPase and downstream effectors, thereby orchestrating the endocytic entry process of NDV. Our findings uncover a novel molecular mechanism of endocytic entry of NDV into host cells and provide novel insight into paramyxovirus mechanisms of entry.
Subject(s)
Macrophages , Newcastle Disease , Newcastle disease virus , Signal Transduction , Virus Internalization , Animals , Endocytosis , Gangliosides/metabolism , Macrophages/metabolism , Macrophages/virology , Newcastle Disease/virology , Newcastle disease virus/physiology , rho GTP-Binding Proteins/metabolismABSTRACT
As a series of our previous studies reported, recombinant yeast can be the oral vaccines to deliver designed protein and DNA, as well as functional shRNA, into dendritic cells (DCs) in mice for specific immune regulation. Here, we report the further optimization of oral yeast-based vaccine from two aspects (yeast characteristics and recombinant DNA constitution) to improve the effect of immune regulation. After screening four genes in negative regulation of glucan synthesis in yeast (MNN9, GUP1, PBS2 and EXG1), this research combined HDR-based genome editing technology with Cre-loxP technology to acquire 15 gene-knockout strains without drug resistance-gene to exclude biosafety risks; afterward, oral feeding experiments were performed on the mice using 15 oral recombinant yeast-based vaccines constructed by the gene-knockout strains harboring pCMV-MSTN plasmid to screen the target strain with more effective inducing mstn-specific antibody which in turn increasing weight gain effect. And subsequently based on the selected gene-knockout strain, the recombinant DNA in the oral recombinant yeast-based vaccine is optimized via a combination of protein fusion expression (OVA-MSTN) and interfering RNA technology (shRNA-IL21), comparison in terms of both weight gain effect and antibody titer revealed that the selected gene-knockout strain (GUP1ΔEXG1Δ) combined with specific recombinant DNA (pCMV-OVA-MSTN-shIL2) had a better effect of the vaccine. This study provides a useful reference to the subsequent construction of a more efficient oral recombinant yeast-based vaccine in the food and pharmaceutical industry.
Subject(s)
DNA, Recombinant , Saccharomyces cerevisiae , Mice , Animals , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , DNA, Recombinant/metabolism , Vaccines, Synthetic , RNA, Small Interfering , Weight GainABSTRACT
Facial soft tissue injuries, often resulting in scarring, pose a challenge in reconstructive and aesthetic surgery due to the need for functional and aesthetic restoration. This study evaluates the efficacy of recombinant human growth factors (rhGFs) in scar remodelling for such injuries. A retrospective evaluation was conducted from January 2020 to January 2023, involving 100 patients with facial soft tissue injuries. Participants were divided equally into a control group, receiving standard cosmetic surgical repair, and an observation group, treated with rhGFs supplemented cosmetic surgery. The study assessed scar characteristics (pigmentation, pliability, vascularity, height), hospital stay duration, tissue healing time, complication rates and patient satisfaction. The observation group demonstrated significant improvements in all scar characteristics, with notably better pigmentation, pliability, vascularity and height compared with the control group. The rhGF treatment also resulted in reduced hospital stay duration and faster tissue healing. Notably, the total complication rate was significantly lower in the observation group (10%) compared with the control group (34%). Additionally, patient satisfaction levels were higher in the observation group, with 98% combined satisfaction compared with 76% in the control group. The application of rhGFs in treating facial soft tissue injuries significantly enhances scar remodelling, expedites healing, reduces complications and improves patient satisfaction. These findings establish rhGFs as a valuable tool in the management of facial soft tissue injuries, highlighting their potential in improving both functional and aesthetic outcomes.
Subject(s)
Facial Injuries , Plastic Surgery Procedures , Soft Tissue Injuries , Humans , Cicatrix/drug therapy , Cicatrix/surgery , Retrospective Studies , Wound Healing , Soft Tissue Injuries/drug therapy , Soft Tissue Injuries/surgery , Facial Injuries/drug therapy , Facial Injuries/surgery , Treatment OutcomeABSTRACT
Glycoprotein non-metastatic melanoma protein B (GPNMB) is ubiquitously expressed and has protective effects on the central nervous system. In particular, it is also expressed in the peripheral nervous system (PNS) and upregulated after peripheral nerve injury. However, the role and underlying mechanism of GPNMB in the PNS, especially in peripheral nerve regeneration (PNR), are still unknown and need to be further investigated. In this study, recombinant human GPNMB (rhGPNMB) was injected into a sciatic nerve injury model. It was found that rhGPNMB facilitated the regeneration and functional recovery of the injured sciatic nerve in vivo. Moreover, it was also confirmed that GPNMB activated the Erk1/2 and Akt pathways via binding with Na+/K + -ATPase α1 (NKA α1) and promoted the proliferation and migration of Schwann cells (SCs) and their expression and secretion of neurotrophic factors and neural adhesion molecules in vitro. Our findings demonstrate that GPNMB facilitates PNR through activation of the Erk1/2 and Akt pathways in SCs by binding with NKA α1 and may be a novel strategy for PNR.
Subject(s)
Melanoma , Peripheral Nerve Injuries , Receptors, Fc , Humans , Proto-Oncogene Proteins c-akt/metabolism , Melanoma/metabolism , Melanoma/pathology , Schwann Cells/metabolism , Nerve Regeneration/physiology , Sciatic Nerve/injuries , Sodium-Potassium-Exchanging ATPase/metabolism , Glycoproteins , Peripheral Nerve Injuries/metabolism , Membrane Glycoproteins/metabolismABSTRACT
SARS-CoV-2 viruses are highly transmissible and immune evasive. It is critical to develop broad-spectrum prophylactic and therapeutic antibodies for potential future pandemics. Here, we used the phage display method to discover nanobodies (Nbs) for neutralizing SARS-CoV-2 viruses especially Omicron strains. The leading nanobody (Nb), namely, Nb4, with excellent physicochemical properties, can neutralize Delta and Omicron subtypes, including BA.1, BA.1.1 (BA.1 + R346K), BA.2, BA.5, BQ.1, and XBB.1. The crystal structure of Nb4 in complex with the receptor-binding domain (RBD) of BA.1 Spike protein reveals that Nb4 interacts with an epitope on the RBD overlapping with the receptor-binding motif, and thus competes with angiotensin-converting enzyme 2 (ACE2) binding. Nb4 is expected to be effective for neutralizing most recent Omicron variants, since the epitopes are evolutionarily conserved among them. Indeed, trivalent Nb4 interacts with the XBB1.5 Spike protein with low nM affinity and competes for ACE2 binding. Prophylactic and therapeutic experiments in mice indicated that Nb4 could reduce the Omicron virus loads in the lung. In particular, in prophylactic experiments, intranasal administration of multivalent Nb4 completely protected mice from Omicron infection. Taken together, these results demonstrated that Nb4 could serve as a potent and broad-spectrum prophylactic and therapeutic Nb for COVID-19.