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Since its invention in the late 1980s, the air-liquid-interface (ALI) culture system has been the standard in vitro model for studying human airway biology and pulmonary diseases. However, in a conventional ALI system, cells are cultured on a porous plastic membrane that is much stiffer than human airway tissues. Here, we develop a gel-ALI culture system by simply coating the plastic membrane with a thin layer of hydrogel with tunable stiffness matching that of healthy and fibrotic airway tissues. We determine the optimum gel thickness that does not impair the transport of nutrients and biomolecules essential to cell growth. We show that the gel-ALI system allows human bronchial epithelial cells (HBECs) to proliferate and differentiate into pseudostratified epithelium. Furthermore, we discover that HBECs migrate significantly faster on hydrogel substrates with stiffness matching that of fibrotic lung tissues, highlighting the importance of mechanical cues in human airway remodeling. The developed gel-ALI system provides a facile approach to studying the effects of mechanical cues in human airway biology and in modeling pulmonary diseases.NEW & NOTEWORTHY In a conventional ALI system, cells are cultured on a plastic membrane that is much stiffer than human airway tissues. We develop a gel-ALI system by coating the plastic membrane with a thin layer of hydrogel with tunable stiffness matching that of healthy and fibrotic airway tissues. We discover that human bronchial epithelial cells migrate significantly faster on hydrogel substrates with pathological stiffness, highlighting the importance of mechanical cues in human airway remodeling.
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Remodelação das Vias Aéreas , Pneumopatias , Humanos , Células Epiteliais , Pulmão , Hidrogéis , Células CultivadasRESUMO
CircRNAs are a member of noncoding RNAs and have been verified to play an important regulatory role in cancers. In CRC, the regulatory mechanisms of various circRNAs have not been elucidated. The expression of circPACRGL and miR-330-3p was detected with qRT-PCR. The protein expression of CDK4, MMP-9, Bcl-2, Bax, cellular nucleic acid-binding protein (CNBP) and ß-actin was measured with western blot. Cell proliferation was analyzed using MTT assay, colony formation assay, and EDU assay. Cell apoptosis was detected using flow cytometry. Cell migration and invasion were measured with wound healing and transwell invasion assay. Luciferase reporter assay and RIP assay was used to determine the relationship of among miR-330-3p, circPACRGL and CNBP in CRC cells. In this study, we found that circPACRGL and CNBP expressed high and miR-330-3p expressed low in CRC tissues and cells. Functional experiments showed that inhibition of circPACRGL reduced cell proliferation, migration and invasion in CRC. In addition, knockdown of circPACRGL contributed to cell apoptosis in CRC. Dual-luciferase report assay determined that circPACRGL was a miR-330-3p sponge molecular and CNBP was a target of miR-330-3p. Reversed experiments showed that the effects of sh-circPACRGL transfection on CRC cells were rescued by up-regulating CNBP expression. In this study, we for the first time found a novel regulatory network of circPACRGL in CRC. The results manifested that circPACRGL affected tumor growth by targeting miR-330-3p/CNBP axis in CRC, highlighting the potential of circPACRGL as a therapeutic target for colorectal cancer.
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Neoplasias Colorretais , MicroRNAs , Humanos , MicroRNAs/genética , MicroRNAs/metabolismo , Linhagem Celular Tumoral , RNA Circular/genética , Neoplasias Colorretais/metabolismo , Movimento Celular , Proliferação de Células , Apoptose , Proteínas de Ligação a RNARESUMO
PURPOSE: Acute respiratory distress syndrome (ARDS), a severe medical condition, is among the major causes of death in critically ill patients. Morphine is used as a therapeutic agent against severe pain. The mechanisms of its reactions over ARDS are not fully understood. The aim of this study was to assess the mechanism of morphine in rats with ARDS. METHODS: Rats were injected with lipopolysaccharide to induce ARDS, and some rats were pre-treated with graded doses of morphine in the lateral ventricles to assess survival and non-infected mortality. Immunohistochemical and HE staining were performed to measure MPO and CD68 activity in the lungs and lung injury. ELISA was conducted to detect the inflammatory factor levels in the plasma and BALF. Co-labeling of µ-opioid receptor (MOR) and c-Fos was observed in the brain tissues. MOR-positive cells in brain tissues were evaluated using immunohistochemistry. The effect of MOR antagonists on ARDS was examined in rats by pre-injection of naloxone or methylnaltrexone. The expression of MyD88, TLR4, and NF-κB was lastly assessed. RESULTS: Dose-independent improvement was observed in respiratory capacity and lung injury in ARDS rats after morphine pre-injection, along with reduced inflammatory factors in the plasma and BALF. MOR-positive cells were elevated after morphine, which occurred within the ventral part of the gigantocellular reticular nucleus (GiV). Naloxone and methylnaltrexone blocked the effects of morphine via central and peripheral MOR. Morphine activated TLR pathway in a MyD88-dependent manner. CONCLUSION: Morphine activates MOR within the GiV and the TLR pathway to attenuate ARDS in rats.
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Lesão Pulmonar , Síndrome do Desconforto Respiratório , Animais , Lipopolissacarídeos , Morfina/farmacologia , Fator 88 de Diferenciação Mieloide , Naloxona/farmacologia , Ratos , Receptores Opioides , Síndrome do Desconforto Respiratório/induzido quimicamente , Síndrome do Desconforto Respiratório/tratamento farmacológicoRESUMO
BACKGROUND: The previous study investigated whether the functions of small, medium, and large high density lipoprotein (S/M/L-HDL) are correlated with protein changes in mice. Herein, the proteomic and functional analyses of high density lipoprotein (HDL) subclasses were performed in humans and rats. METHODS: After purifying S/M/L-HDL subclasses from healthy humans (n = 6) and rats (n = 3) using fast protein liquid chromatography (FPLC) with calcium silica hydrate (CSH) resin, the proteomic analysis by mass spectrometry was conducted, as well as the capacities of cholesterol efflux and antioxidation was measured. RESULTS: Of the 120 and 106 HDL proteins identified, 85 and 68 proteins were significantly changed in concentration among the S/M/L-HDL subclasses in humans and rats, respectively. Interestingly, it was found that the relatively abundant proteins in the small HDL (S-HDL) and large HDL (L-HDL) subclasses did not overlap, both in humans and in rats. Next, by searching for the biological functions of the relatively abundant proteins in the HDL subclasses via Gene Ontology, it was displayed that the relatively abundant proteins involved in lipid metabolism and antioxidation were enriched more in the medium HDL (M-HDL) subclass than in the S/L-HDL subclasses in humans, whereas in rats, the relatively abundant proteins associated with lipid metabolism and anti-oxidation were enriched in M/L-HDL and S/M-HDL, respectively. Finally, it was confirmed that M-HDL and L-HDL had the highest cholesterol efflux capacity among the three HDL subclasses in humans and rats, respectively; moreover, M-HDL exhibited higher antioxidative capacity than S-HDL in both humans and rats. CONCLUSIONS: The S-HDL and L-HDL subclasses are likely to have different proteomic components during HDL maturation, and results from the proteomics-based comparison of the HDL subclasses may explain the associated differences in function.
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Antioxidantes , Proteômica , Humanos , Ratos , Camundongos , Animais , Estudo de Prova de Conceito , Lipoproteínas HDL , ColesterolRESUMO
The use of proteins as biomolecular templates to synthesize atomically precise metal nanoclusters has been gaining traction due to their appealing properties such as photoluminescence, good colloidal- and photostability and biocompatibility. The synergistic effect of using a protein scaffold and metal nanoclusters makes it especially attractive for biomedical applications. Unlike other reviews, we focus on proteins in general as the protective ligand for various metal nanoclusters and highlight their applications in the biomedical field. We first introduce the approaches and underlined principles in synthesizing protein-templated metal nanoclusters and summarize some of the typical proteins that have been used thus far. Afterwards, we highlight the key physicochemical properties and the characterization techniques commonly used for the size, structure and optical properties of protein-templated metal nanoclusters. We feature two case studies to illustrate the importance of combining these characterization techniques to elucidate the formation process of protein-templated metal nanoclusters. Lastly, we highlight the promising applications of protein-templated metal nanoclusters in three areas-biosensing, diagnostics and therapeutics.
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Ouro , Nanopartículas Metálicas , Ouro/química , Biofarmácia , Proteínas , Nanopartículas Metálicas/químicaRESUMO
PURPOSE: To compare the impact of different statins therapies on the reduction of carotid intima-media thickness (CIMT) may reflect their cardiovascular benefits which is useful in clinical decision. METHODS: PubMed, EMBASE, Cochrane Library, and Web of Science were searched, and 3539 articles published from 1992 to 2020 were retrieved. CIMT in randomized controlled trials for statins therapies were included for traditional and network meta-analyses analyzed by Stata 16. The quality of included studies was assessed by the Cochrane Collaboration's tool. RESULTS: Thirty-three randomized controlled trials (n=8762) were eligible for network meta-analysis, of which 18 randomized controlled trials (n=5252) were included for comparison between statins and no statins and 11 randomized controlled trials (n=1338) were included for comparison between high-intensity statins or combination with niacin/ezetimibe and moderate/low-intensity statins in 2 traditional meta-analyses. In the traditional meta-analyses, the statins groups significantly reduce CIMT compared to no statins (standard mean difference=-0.207, 95% confidence interval: -0.291 to -0.123, p<0.001), while high-intensity statins or combination with niacin/ezetimibe performed significant CIMT reduction compared to moderate/low-intensity statins (standard mean difference=-0.287, 95% confidence interval: -0.460 to -0.114, p=0.001). In the network meta-analysis, a relative rank for the ability to reduce CIMT was given as follows: combination therapy with niacin (mean rank: 1.7), high-intensity statins, combination therapy with ezetimibe, and moderate/low-intensity statins. CONCLUSION: Statins combined with niacin performed a greater CIMT reduction compared to high-intensity statins alone and combination therapies with ezetimibe. The advantage of niacin-combined statins therapies to improve cardiovascular endpoint needs further validation through randomized controlled trials. CLINICAL TRIAL REGISTRATION: PROSPERO, CRD42020175972.
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Espessura Intima-Media Carotídea , Ezetimiba/uso terapêutico , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Hipolipemiantes/uso terapêutico , Niacina/uso terapêutico , Quimioterapia Combinada , Ezetimiba/administração & dosagem , Humanos , Inibidores de Hidroximetilglutaril-CoA Redutases/administração & dosagem , Hipolipemiantes/administração & dosagem , Metanálise em Rede , Niacina/administração & dosagem , Ensaios Clínicos Controlados Aleatórios como AssuntoRESUMO
Increased protein synthesis during cell proliferation is accompanied by a compensatory increase in efficient ribosome production, but the mechanisms by which cells adapt to this requirement are not fully understood. In the present study, we demonstrate evidence that Pygo (Pygopus), a protein originally identified as a core component of the Wnt-ß-catenin transcription complex is also involved in rRNA transcription during cancer cell growth. Pygo was detected in the nucleoli of several transformed cell lines and was associated with treacle and UBF (upstream binding factor), proteins that are essential for ribosome biogenesis in development and cancer. Pygo was also detected at the ribosomal gene promoter along with core components of the rDNA (ribosomal DNA) transcription complex. RNAi (RNA interference)-mediated depletion of hPygo2 (human Pygo 2) reduced histone H4 acetylation at the rDNA promoter, down-regulated rRNA production, and induced growth arrest in both p53-positive and -negative cells. In p53-positive cells, hPygo2 knockdown triggered the ribosomal stress pathway, culminating in p53-dependent growth arrest at G1-phase of the cell cycle. The results of the present study suggest a novel involvement of Pygo in the promotion of rRNA transcription in cancer cells.
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Peptídeos e Proteínas de Sinalização Intracelular/fisiologia , Transcrição Gênica/efeitos dos fármacos , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Nucléolo Celular/metabolismo , Células HeLa , Histona Acetiltransferases/metabolismo , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , RNA Ribossômico/genéticaRESUMO
Pulmonary drug delivery is critical for the treatment of respiratory diseases. However, the human airway surface presents multiple barriers to efficient drug delivery. Here, we report a bottlebrush poly(ethylene glycol) (PEG-BB) nanocarrier that can translocate across all barriers within the human airway surface. Guided by a molecular theory, we design a PEG-BB molecule consisting of a linear backbone densely grafted by many (â¼1000) low molecular weight (â¼1000 g/mol) polyethylene glycol (PEG) chains; this results in a highly anisotropic, wormlike nanocarrier featuring a contour length of â¼250 nm, a cross-section of â¼20 nm, and a hydrodynamic diameter of â¼40 nm. Using the classic air-liquid-interface culture system to recapitulate essential biological features of the human airway surface, we show that PEG-BB rapidly penetrates through endogenous airway mucus and periciliary brush layer (mesh size of 20-40 nm) to be internalized by cells across the whole epithelium. By quantifying the cellular uptake of polymeric carriers of various molecular architectures and manipulating cell proliferation and endocytosis pathways, we show that the translocation of PEG-BB across the epithelium is driven by bottlebrush architecture-enhanced endocytosis. Our results demonstrate that large, wormlike bottlebrush PEG polymers, if properly designed, can be used as a carrier for pulmonary and mucosal drug delivery.
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Portadores de Fármacos , Endocitose , Polietilenoglicóis , Humanos , Polietilenoglicóis/química , Portadores de Fármacos/química , Nanopartículas/química , Mucosa Respiratória/metabolismo , Mucosa Respiratória/citologia , Tamanho da Partícula , Sistemas de Liberação de Medicamentos , Proliferação de Células/efeitos dos fármacosRESUMO
Pulmonary drug delivery is critical to the treatment of respiratory diseases. However, the human airway surface presents multiscale barriers to efficient drug delivery. Here we report a bottlebrush polyethylene glycol (PEG-BB) nanocarrier that can translocate across all barriers within the human airway surface. Guided by the molecular theory, we design a PEG-BB molecule consisting of a linear backbone densely grafted by many (â¼1,000) low molecular weight (â¼1000 g/mol) PEG chains; this results in a highly anisotropic, wormlike nanocarrier featuring a contour length of â¼250 nm, a cross-section of â¼20 nm, and a hydrodynamic diameter of â¼40 nm. Using the classic air-liquid-interface culture system to recapitulate essential biological features of the human airway surface, we show that PEG-BB rapidly penetrates through endogenous airway mucus and periciliary brush layer (mesh size of 20-40 nm) to be internalized by cells across the whole epithelium. By quantifying the cellular uptake of polymeric carriers of various molecular architectures and manipulating cell proliferation and endocytosis pathways, we show that the translocation of PEG-BB across the epithelium is driven by bottlebrush architecture enhanced endocytosis. Our results demonstrate that large, wormlike bottlebrush PEG polymers, if properly designed, can be used as a novel carrier for pulmonary and mucosal drug delivery.
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Cantharidin, a terpenoid produced by blister beetles, has been used in traditional Chinese medicine to treat various ailments and cancers. However, its biological activity, impact, and anticancer mechanisms remain unclear. The Cantharidin chemical gene connections were identified using various databases. The GSE21815 dataset was used to collect the gene expression information. Differential gene analysis and gene ontology analyses were performed. Gene set enrichment analysis was used to assess the activation of disease pathways. Weighted gene co-expression network analysis and differential analysis were used to identify illness-associated genes, examine differential genes, and discover therapeutic targets via protein-protein interactions. MCODE analysis of major subgroup networks was used to identify critical genes influenced by Cantharidin, examine variations in the expression of key clustered genes in colorectal cancer vs. control samples, and describe the subject operators. Single-cell GSE188711 dataset was preprocessed to investigate Cantharidin's therapeutic targets and signaling pathways in colorectal cancer. Single-cell RNA sequencing was utilized to identify 22 cell clusters and marker genes for two different cell types in each cluster. The effects of different Cantharidin concentrations on colorectal cancer cells were studied in vitro. One hundred and ninety-seven Cantharidin-associated target genes and 480 critical genes implicated in the development of the illness were identified. Cantharidin significantly inhibited the proliferation and migration of HCT116 cells and promoted apoptosis at certain concentrations. Patients on current therapy develop inherent and acquired resistance. Our study suggests that Cantharidin may play an anti-CRC role by modulating immune function.
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Carbon dioxide phase transition fracturing (CDPTF) is widely regarded as a promising coal seam mining technique because it can effectively improve coal seam permeability and prevent gas outbursts. An impact pressure test system of CDPTF was developed, and the effects of different factors on impact pressure were investigated by combining CO2 release experiments and smoothed particle hydrodynamics numerical simulation. In addition, based on the Peng-Robinson equation and the pipeline pressure drop formula, new mathematical models for the pressure equation in the buffer tank and the velocity of gaseous CO2 at the nozzle were established. The results show that the impact pressure of CDPTF can be divided into rapid boost, fluctuation, and attenuation stages. The impact distance and impact angle have the most significant effects on pressure. The models of the pressure in the buffer tank and the velocity of gaseous CO2 at the nozzle well-simulated the experimentally obtained impact pressure curves. The research results could provide a reference for the loading study of CDPTF.
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Introduction: Background and aims: although sarcopenia is associated with several types of cancer, there is limited research regarding its effect on breast cancer. We aimed to explore the causality between sarcopenia-related traits and the incidence and prognosis of breast cancer. Methods: two-sample bidirectional and multivariate Mendelian randomization (MR) analyses were utilized in this study. Genome-wide association studies were used to genetically identify sarcopenia-related traits, such as appendicular lean mass, grip strength of both hands, and walking pace. Data on the incidence and prognosis of breast cancer were collected from two extensive cohort studies. Multivariate MR analysis was used to adjust for body mass index, waist circumference, and whole-body fat mass. The primary method used for analysis was inverse-variance weighted analysis. Results: a significant association was found between appendicular lean mass and ER- breast cancer (OR = 0.873, 95 % CI: 0.817-0.933, p = 6.570 × 10-5). Increased grip strength of the left hand was associated with a reduced risk of ER- breast cancer (OR = 0.744, 95 % CI: 0.579-0.958, p = 0.022). Stronger grip strength of the right hand was associated with prolonged survival time of ER+ breast cancer patients (OR = 0.463, 95 % CI: 0.242-0.882, p = 0.019). In the multivariable MR analysis, appendicular lean mass, grip strength of both hands, and walking pace were still genetically associated with the development of total breast cancer and ER-/+ breast cancer. Conclusions: several sarcopenia-related traits were genetically associated with the occurrence and prognosis of breast cancer. It is crucial for elderly women to increase their strength and muscle mass to help prevent breast cancer.
Introducción: Antecedentes y objetivos: aunque la sarcopenia se asocia a múltiples tipos de cáncer, los estudios sobre sus efectos sobre el cáncer de mama son limitados. Nuestro objetivo es explorar la relación causal entre las características relacionadas con la sarcopenia y la incidencia y el pronóstico del cáncer de mama. Método: este estudio utilizó un análisis de aleatorización mendeliana (MR) bidireccional y multivariable de doble muestra. Los estudios de asociación genómica completa se utilizan para identificar genéticamente características relacionadas con la sarcopenia, como la masa magra apendicular, la fuerza de agarre de las manos y la velocidad al caminar. Los datos de incidencia y pronóstico del cáncer de mama provienen de dos amplios estudios de cohortes. El análisis de MR multivariable se utilizó para ajustar el índice de masa corporal, la circunferencia de la cintura y la masa grasa corporal total. El principal método de análisis fue el análisis ponderado por ANOVA inverso. Resultados: la masa magra apendicular se asoció significativamente al cáncer de mama ER- (OR = 0,873, IC 95 %: 0,817-0,933, p = 6,570 × 10-5), el aumento de la fuerza de agarre del lado izquierdo se asoció a una disminución del riesgo de cáncer de mama ER- (OR = 0,744, IC 95 %: 0,579-0,958, p = 0,022) y el aumento de la fuerza de agarre del lado derecho se asoció a una mayor supervivencia de los pacientes con cáncer de mama ER+ (OR = 0,463, IC 95 %: 0,24-0,882, P = 0,019). En el análisis MR multivariable, la masa magra apendicular, la fuerza de agarre de ambas manos y la velocidad al caminar mantuvieron su asociación genética con la aparición del cáncer de mama total y del cáncer de mama ER-/+. Conclusión: varios rasgos relacionados con la sarcopenia tienen correlación genética con la aparición y el pronóstico del cáncer. Mejorar la fuerza y la masa muscular de las mujeres mayores es fundamental para ayudar a prevenir el cáncer de mama.
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Neoplasias da Mama , Força da Mão , Análise da Randomização Mendeliana , Sarcopenia , Humanos , Neoplasias da Mama/genética , Sarcopenia/genética , Sarcopenia/epidemiologia , Feminino , Prognóstico , Pessoa de Meia-Idade , Estudo de Associação Genômica Ampla , Idoso , Estudos de Coortes , IncidênciaRESUMO
Background: Single-branched stent grafts and the chimney technique are widely used in the treatment of type B aortic dissection (TBAD). The main objective of this study was to compare the outcomes of single-branched stent grafts and the chimney technique in the treatment of TBAD. Methods: From January 2019 to December 2021, the retrospective cohort study contained a cohort of 91 patients with TBAD undergoing thoracic endovascular aortic repair (TEVAR) using single-branched stent grafts and the chimney technique. Group A included 55 patients treated with single-branched covered stents, and Group B included 36 patients treated with the chimney technique. We compared the effects of the procedures on peri-/post-operative outcomes between the two groups. The primary endpoint is clinical death, and the secondary endpoints include the patency of branch stents, the incidence of cerebral infarction, false lumen thrombosis, and the proportion of paraplegia. Results: For the baseline data, the two groups of patients show no differences in terms of age, gender, and associated symptoms. All procedures were successfully performed in both groups. The median follow-up period was 17.6 months (range, 10-34 months). During TEVAR, 5 (9.1%) type I endoleaks occurred in group A, and 11 (30.6%) occurred in group B (P<0.05). During follow-up, there were 2 cases (3.6%) of paraplegia and 1 case (1.8%) of cerebral infarction in Group A, while Group B had 1 case (2.8%) of paraplegia. Three patients in group B reported retrograde type A aortic dissection (RTAD), and 1 of them died (2.8%); however, there were no RTAD cases in group A. Complete thrombosis of the false lumen in the thoracic aorta was observed in 45.5% (25/55) of patients in group A and in 41.7% (15/36) in group B (P=0.72). No significant difference in the thrombosis-volume ratio in the whole false lumen was found during follow-up between group A (81.0%±2.9%) and group B (81.8%±2.6%; P=0.23). Conclusions: Branched stent grafts can be used in cases with insufficient proximal landing zones and reduce the occurrence of type 1 endoleak compared to the chimney technique. This may help to prevent RTAD. Further research, including more cases and longer follow-up periods, is needed to substantiate these results.
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[This corrects the article DOI: 10.1371/journal.pone.0061852.].
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The Amur tiger is currently confronted with challenges of anthropogenic development, leading to its population becoming fragmented into two geographically isolated groups: smaller and larger ones. Small and isolated populations frequently face a greater extinction risk, yet the small tiger population's genetic status and survival potential have not been assessed. Here, a total of 210 samples of suspected Amur tiger feces were collected from this small population, and the genetic background and population survival potentials were assessed by using 14 microsatellite loci. Our results demonstrated that the mean number of alleles in all loci was 3.7 and expected heterozygosity was 0.6, indicating a comparatively lower level of population genetic diversity compared to previously reported studies on other subspecies. The genetic estimates of effective population size (Ne) and the Ne/N ratio were merely 7.6 and 0.152, respectively, representing lower values in comparison to the Amur tiger population in Sikhote-Alin (the larger group). However, multiple methods have indicated the possibility of genetic divergence within our isolated population under study. Meanwhile, the maximum kinship recorded was 0.441, and the mean inbreeding coefficient stood at 0.0868, both of which are higher than those observed in other endangered species, such as the African lion and the grey wolf. Additionally, we have identified a significant risk of future extinction if the lethal equivalents were to reach 6.26, which is higher than that of other large carnivores. Further, our simulation results indicated that an increase in the number of breeding females would enhance the prospects of this population. In summary, our findings provide a critical theoretical basis for further bailout strategies concerning Amur tigers.
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Leões , Tigres , Animais , Feminino , Tigres/genética , Espécies em Perigo de Extinção , Heterozigoto , Densidade Demográfica , Repetições de Microssatélites/genética , Leões/genética , Conservação dos Recursos Naturais , Variação GenéticaRESUMO
BACKGROUND: The effects of hypoxia conditioning, which involves recurrent exposure to hypoxia combined with exercise training, on improving body composition in the ageing population have not been extensively investigated. OBJECTIVE: This meta-analysis aimed to determine if hypoxia conditioning, compared to similar training near sea level, maximizes body composition benefits in middle-aged and older adults. METHODS: A literature search of PubMed, EMBASE, Web of Science, Scopus and CNKI (China National Knowledge Infrastructure) databases (up to 27th November 2022) was performed, including the reference lists of relevant papers. Three independent reviewers extracted study characteristics and health outcome measures. Search results were limited to original studies of the effects of hypoxia conditioning on body composition in middle-aged and older adults. RESULTS: Twelve studies with a total of 335 participants were included. Hypoxia conditioning induced greater reductions in body mass index (MD = -0.92, 95%CI: -1.28 to -0.55, I2 = 0%, p < 0.00001) and body fat (SMD = -0.38, 95%CI: -0.68 to -0.07, I2 = 49%, p = 0.01) in middle-aged and older adults compared with normoxic conditioning. Hypoxia conditioning improved lean mass with this effect not being larger than equivalent normoxic interventions in either middle-aged or older adults (SMD = 0.07, 95%CI -0.12 to 0.25, I2 = 0%, p = 0.48). Subgroup analysis showed that exercise in moderate hypoxia (FiO2 > 15%) had larger effects than more severe hypoxia (FiO2 ≤ 15%) for improving body mass index in middle-aged and older adults. Hypoxia exposure of at least 60 min per session resulted in larger benefits for both body mass index and body fat. CONCLUSION: Hypoxia conditioning, compared to equivalent training in normoxia, induced greater body fat and body mass index improvements in middle-aged and older adults. Adding hypoxia exposure to exercise interventions is a viable therapeutic solution to effectively manage body composition in ageing population.
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OBJECTIVES: This study conducted integrated analysis of bulk RNA sequencing, single-cell RNA sequencing and Weighted Gene Co-expression Network Analysis (WGCNA), to comprehensively decode the most essential genes of intervertebral disc degeneration (IDD); then mainly focused on the JAK3 macromolecule to identify natural compounds to provide more candidate drug options in alleviating IDD. METHODS: In the first part, we performed single-cell transcriptome analysis and WGCNA workflow to delineate the most pivotal genes of IDD. Then series of structural biology approaches and high-throughput virtual screening techniques were performed to discover potential compounds targeting JAK-STAT signaling pathway, such as Libdock, ADMET, precise molecular docking algorithm and in-vivo drug stability assessment. RESULTS: Totally 4 hub genes were determined in the development of IDD, namely VEGFA, MMP3, TNFSF11, and TIMP3, respectively. Then, 3 novel natural materials, ZINC000014952116, ZINC000003938642 and ZINC000072131515, were determined as potential compounds, with less toxicities and moderate ADME characteristics. In-vivo drug stability assessment suggested that these drugs could interact with JAK3, and their ligand-JAK3 complexes maintained the homeostasis in-vivo, which acted as regulatory role to JAK3 protein. Among them, ZINC000072131515, also known as Menaquinone, demonstrated significant protective roles to alleviate the progression of IDD in vitro, which proved the nutritional therapy in alleviating IDD. CONCLUSIONS: This study reported the essential genes in the development of IDD, and also the roles of Menaquinone to ameliorate IDD through inhibiting JAK3 protein. This study also provided more options and resources on JAK3 targeted screening, which may further expand the drug resources in the pharmaceutical market.
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Degeneração do Disco Intervertebral , Janus Quinase 3 , Humanos , Biologia , Degeneração do Disco Intervertebral/tratamento farmacológico , Degeneração do Disco Intervertebral/genética , Degeneração do Disco Intervertebral/metabolismo , Janus Quinase 3/genética , Simulação de Acoplamento Molecular , Transcriptoma , Vitamina K 2RESUMO
Intervertebral disc degeneration (IDD) is a progressive and chronic disease, the mechanisms have been studied extensively as a whole, while the cellular heterogeneity of cells in nucleus pulposus (NP) tissues remained controversial for a long time. This study conducted integrated analysis through single-cell sequencing analysis, weighted gene co-expression network analysis (WGCNA), and differential expression analysis, to systematically decipher the longitudinal alterations of distinct NP subtypes, and also analyzed the most essential genes in the development of IDD. Then, this study further conducted structural biology method to discover the potential lead compounds through a suite of advanced approaches like high-throughput screening (HTVS), pharmaceutical characteristics assessment, CDOCKER module as well as molecular dynamics simulation, etc., aiming to ameliorate the progression of IDD. Totally 5 NP subpopulations were identified with distinct biological functions based on their unique gene expression patterns. The predominant dynamics changes mainly involved RegNPs and EffNPs, the RegNPs were mainly aggregated in normal NP tissues and drastically decreased in degenerative NP, while EffNPs, as pathogenic subtype, exhibited opposite phenomenon. Importantly, this study further reported the essential roles of Menaquinone in alleviating degenerative NP cells for the first time, which could provide solid evidence for the application of nutritional therapy in the treatment of IDD. This study combined scRNA-seq, bulk-RNA seq and HTVS techniques to systematically decipher the longitudinal changes of NP subtypes during IDD. EffNPs were considered to be 'chief culprit' in IDD progression, while the novel natural drug Menaquinone could reverse this phenomenon.Communicated by Ramaswamy H. Sarma.
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Intervertebral disk degeneration (IDD) is a chronic inflammatory disease with intricate connections between immune infiltration and oxidative stress (OS). Complex cell niches exist in degenerative intervertebral disk (IVD) and interact with each other and regulate the disk homeostasis together. However, few studies have used longitudinal approach to describe the immune response of IDD progression. Here, we conducted conjoint analysis of bulk-RNA sequencing and single-cell sequencing, together with a series of techniques like weighted gene co-expression network analysis (WGCNA), immune infiltration analysis, and differential analysis, to systematically decipher the difference in OS-related functions of different cell populations within degenerative IVD tissues, and further depicted the longitudinal alterations of immune cells, especially monocytes/macrophages in the progression of IDD. The OS-related genes CYP1A1, MMP1, CCND1, and NQO1 are highly expressed and might be diagnostic biomarkers for the progression of IDD. Further landscape of IVD microenvironment showed distinct changes in cell proportions and characteristics at late degeneration compared to early degeneration of IDD. Monocytes/macrophages were classified into five distinct subpopulations with different roles. The trajectory lineage analysis revealed transcriptome alterations from effector monocytes/macrophages and regulatory macrophages to other subtypes during the evolution process and identified monocytes/macrophage subpopulations that had rapidly experienced the activation of inflammatory or anti-inflammatory responses. This study further proposed that personalized therapeutic strategies are needed to be formulated based on specific monocyte/macrophage subtypes and degenerative stages of IDD.