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PURPOSE: Urgent start peritoneal dialysis (USPD) is an effective therapeutic method for end-stage renal disease (ESRD). However, whether it is safe to initiate peritoneal dialysis (PD) within 24 h unclear. We examined the short-term outcomes of a break-in period (BI) of 24 h for patients undergoing USPD. METHODS: This real-world, multicenter, retrospective cohort study evaluated USPD patients from five centers from January 2013 to August 2020. Patients were divided into BI ≤ 24 h or BI > 24 h groups. The Primary outcomes included incidence of mechanical and infectious complications. The secondary outcome was technique failure. Moreover, we presented a subgroup analysis for patients who did not receive temporary hemodialysis (HD). RESULTS: A total of 871 USPD patients were included: 470 in the BI ≤ 24 h and 401 in the BI > 24 h groups. Mechanical and infectious complications did not differ between the two groups across the follow-up timepoints (2 weeks, 1 month, 3 months, and 6 months) (p > 0.05). Multiple logistic regression analysis revealed that BI ≤ 24 h was not an independent risk factor for mechanical complications, catheter migration, or infectious complications (p > 0.05). A BI ≤ 24 h was not an independent significant risk factor for technique failure by multivariate Cox regression analysis (p > 0.05). The subgroup analysis of patients who did not receive temporary HD returned the same results. CONCLUSION: Initiating PD within 24 h of catheter insertion was not associated with increased mechanical complications, infectious complications, or technique failures.
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Fallo Renal Crónico/terapia , Diálisis Peritoneal/efectos adversos , Diálisis Peritoneal/métodos , Adulto , China , Estudios de Factibilidad , Femenino , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Complicaciones Posoperatorias/epidemiología , Prevalencia , Estudios Retrospectivos , Factores de Riesgo , Factores de TiempoRESUMEN
Modifying the side chain of poly(meth)acrylate with moieties originating from biocompatible polymers can be an effective method for developing novel blood-compatible polymers. Inspired by biocompatible poly(2-methyl-2-oxazoline) (PMeOx) and poly(2-ethyl-2-oxazoline) (PEtOx), four water-soluble poly(tertiary amide acylate) analogues bearing a pendant tertiary amide were synthesized. The results of hemolysis and cell viability tests showed that all the poly(tertiary amide acylate) analogues were compatible with red blood cells, HeLa cells, and normal human dermal fibroblasts as PMeOx or PEtOx. Among the four poly(tertiary amide acylate) analogues, poly[2-(N-methylpropionamido)ethyl acrylate] (PPEA) and poly[2-(N-ethylacetamido)ethyl acrylate] (PEAE) showed thermosensitivity in aqueous solution; especially, PPEA (10 mg mL-1) exhibited a lower critical solution temperature of 37 °C. Water-insoluble copolymers were prepared to investigate the possibility of applying these synthesized polymers as blood-compatible coatings. The poly[n-butyl methacrylate70-co-2-(N-methylacetamido)ethyl methacrylate30] (coPAEM) coatings significantly suppressed plasma protein adsorption, denaturation degree of adsorbed fibrinogen, and platelet adhesion. Intermediate water (IW), whose content can generally indicate the blood compatibility of polymers, was found in all hydrated homopolymers and copolymers by differential scanning calorimetry. The present work demonstrated that the tertiary amide moiety in the side chain of poly(meth)acrylate was an effective contributor to blood compatibility and IW.
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Amidas , Materiales Biocompatibles , Acrilatos , Células HeLa , Humanos , Oxazoles , Polímeros , AguaRESUMEN
Upregulated expression of P-selectin on activated endothelium and platelets significantly contributes to the initiation and progression of vaso-occlusive crises (VOC), a major cause of morbidity in sickle cell disease (SCD). Crizanlizumab (ADAKVEO®), a humanized monoclonal antibody against P-selectin, primarily inhibits the interaction between leukocytes and P-selectin, and has been shown to decrease the frequency of VOCs in clinical trials. However, the lack of reliable in vitro assays that objectively measure leukocyte adhesion to P-selectin remains a critical barrier to evaluating and improving the therapeutic treatment in SCD. Here, we present a standardized microfluidic BioChip whole blood adhesion assay to assess leukocyte adhesion to P-selectin under physiologic flow conditions. Our results demonstrated heterogeneous adhesion by leukocytes to immobilized P-selectin, and dose-dependent inhibition of this adhesion following pre-exposure to Crizanlizumab. Importantly, treatment with Crizanlizumab following adhesion to P-selectin promoted detachment of rolling, but not of firmly adherent leukocytes. Taken together, our results suggest that the microfluidic BioChip system is a promising in vitro assay with which to screen patients, monitor treatment response, and guide current and emerging anti-adhesive therapies in SCD.
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Anemia de Células Falciformes/tratamiento farmacológico , Anticuerpos Monoclonales Humanizados/farmacología , Adhesión Celular/efectos de los fármacos , Leucocitos/efectos de los fármacos , Selectina-P/antagonistas & inhibidores , Adulto , Anciano , Femenino , Humanos , Dispositivos Laboratorio en un Chip/normas , Leucocitos/citología , Masculino , Técnicas Analíticas Microfluídicas/instrumentación , Técnicas Analíticas Microfluídicas/métodos , Técnicas Analíticas Microfluídicas/normas , Persona de Mediana Edad , Adulto JovenRESUMEN
Sickle cell disease (SCD) is a recessive genetic blood disorder exhibiting abnormal blood rheology. Polymerization of sickle hemoglobin, due to a point mutation in the ß-globin gene of hemoglobin, results in aberrantly adhesive and stiff red blood cells (RBCs). Hemolysis, abnormal RBC adhesion, and abnormal blood rheology together impair endothelial health in people with SCD, which leads to cumulative systemic complications. Here, we describe a microfluidic assay combined with a micro particle image velocimetry technique for the integrated in vitro assessment of whole blood viscosity (WBV) and RBC adhesion. We examined WBV and RBC adhesion to laminin (LN) in microscale flow in whole blood samples from 53 individuals with no hemoglobinopathies (HbAA, N = 10), hemoglobin SC disease (HbSC, N = 14), or homozygous SCD (HbSS, N = 29) with mean WBV of 4.50 cP, 4.08 cP, and 3.73 cP, respectively. We found that WBV correlated with RBC count and hematocrit in subjects with HbSC or HbSS. There was a significant inverse association between WBV and RBC adhesion under both normoxic and physiologically hypoxic (SpO2 of 83%) tests, in which lower WBV associated with higher RBC adhesion to LN in subjects with HbSS. Low WBV has been found by others to associate with endothelial activation. Altered WBV and abnormal RBC adhesion may synergistically contribute to the endothelial damage and cumulative pathophysiology of SCD. These findings suggest that WBV and RBC adhesion may serve as clinically relevant biomarkers and endpoints in assessing emerging targeted and curative therapies in SCD.
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Anemia de Células Falciformes/sangre , Viscosidad Sanguínea , Adhesión Celular , Eritrocitos Anormales/metabolismo , Biomarcadores/sangre , Femenino , Humanos , MasculinoRESUMEN
BACKGROUND The retropancreatic fusion fascia is recognized as the anatomical landmark during retropancreatic mobilization. However, its role in D2 total gastrectomy for proximal advanced gastric cancer (PAGC) remains unexplored. This study aimed to develop the retropancreatic fusion fascia-oriented ex vivo approach for splenic hilar lymphadenectomy (RP-SL) to avoid difficulty in using the total laparoscopic approach (TL-SL). MATERIAL AND METHODS The data for patients with PAGC who underwent D2 total gastrectomy were retrieved from our clinical database and electronic medical records (December 2016 to December 2018), with a 1:1 match ratio for balance of the 2 groups. RESULTS In sum, 84 matched patients were included in the study. There were 2360 retrieved lymph nodes (LNs), with an average of 28.10. Sixteen patients were confirmed with positive splenic hilar lymphadenectomy, and the mean harvested lymph nodes (LNs) were significantly increased in the RP-SL group compared to the TL-SL group (3.07 vs. 2.29, P<0.001), decreased operative time (193.21 min vs. 247.74 min, P<0.001), and less blood loss (96.90 mL vs. 185.24 mL, P=0.001) in the RP-SL group. Postoperative hospital stay (6.55 days vs. 7.26 days), rate of morbidity (9.50% vs. 11.91%), and overall costs (¥65255.64 vs. ¥64419.91) were comparable between the groups (P>0.05). CONCLUSIONS The landmark at the conjunction between the superior mesenteric vessels and the inferior pancreatic margin made it feasible to identify the retropancreatic areolar. The RP-SL approach was safe and efficient for splenic hilar lymphadenectomy.
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Puntos Anatómicos de Referencia/anatomía & histología , Gastrectomía/métodos , Neoplasias Gástricas/cirugía , China , Fascia/patología , Tiempo de Internación , Escisión del Ganglio Linfático/métodos , Ganglios Linfáticos/patología , Tempo Operativo , Estudios Retrospectivos , Bazo/patología , Neoplasias Gástricas/patologíaRESUMEN
There are many factors to consider in the field of tissue engineering. For articular cartilage repair, this includes seed cells, scaffolds and chondrotrophic hormones. This review primarily focuses on the seed cells and scaffolds. Extracellular matrix proteins provide a natural scaffold for cell attachment, proliferation and differentiation. The structure and composition of tissue-derived scaffolds and native tissue are almost identical. As such, tissue-derived scaffolds hold great promise for biomedical applications. However, autologous tissue-derived scaffolds also have many drawbacks for transplantation, as harvesting autografts is limited to available donor sites and requires secondary surgery, therefore imparting additional damage to the body. This review summarizes and analyzes various cell sources and tissue-derived scaffolds applied in orthopedic tissue engineering.
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Cartílago Articular/fisiología , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Animales , Cartílago Articular/citología , HumanosRESUMEN
The use of a resin to selectively separate thiomolybdate from a tungsten (W) feed solution is a well-known protocol for achieve high-purity W products; however, the regeneration of saturated resin is laborious. In this study, poly(diallyl dimethyl ammonium chloride) (PDADMA) was used to modify ultrasound-pretreated montmorillonite (Mt) for W and molybdenum (Mo) separation for the first time, and the resultant tetrathiomolybdate (MoS42-)-loaded composite was further tested to remove heavy metals instead of regeneration. Among the three variables of ultrasound pretreatment, that is, Mt concentration, ultrasound power, and treatment time, the Mt concentration exhibited the most significant influence followed by ultrasound power on the separation performance of W and Mo. Compared to the distance of the interlayer space and the surface charge of the modified Mt, the PDADMA content showed a closer correlation with the W/Mo separation coefficient. Assisted by Box-Behnken design, with Mt concentration of 6.9 g/L, ultrasound power of 593.8 W, and treatment time of 13.8 min, the composite with the greatest separation coefficient was obtained. The adsorption of Cu(II) on the optimal W/Mo separation-derived composite was ascribed to the formation of Cu-S complexes, while that of Pb(II) was attributed to complexation and surface precipitation. In contrast, ion exchange with the initially loaded anions, reduction by sulfide to Cr(III), and formation of Cr(III)-S complexes accounted for Cr(VI) removal. The adsorption of Cu(II) and Pb(II) equilibrated faster and showed higher acid-resistance than that of Cr(VI). The adsorption capacities for Cu(II), Pb(II), and Cr(VI) were 0.535, 1.398, and 0.882 mmol/g, respectively. Applying PDADMA to modify Mt as a reagent for W/Mo separation was feasible, and the derived composite was capable of removing cationic and anionic heavy metals.
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Drug-eluting balloon (DEB) angioplasty has emerged as an effective treatment for cardiovascular and cerebrovascular diseases. However, distal embolism and late lumen restenosis could be caused by drug loss during DEB handling and rapid drug metabolization. Here, a drug-loaded balloon equipped with tip-separable microneedles on the balloon surface (MNDLB) was developed. Inbuilt near-infrared (NIR) ring laser inside the catheter inner shaft was introduced to activate the biodegradable microneedle tips for the first time. The drug-loaded tips thus could be embedded in the vasculature and then released antiproliferative drug - paclitaxel slowly via polymer degradation for more than half a year. A significant increase in drug delivery efficiency and superior therapeutic effectiveness compared with the standard DEB were demonstrated using an atherosclerosis rabbit model.
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INTRODUCTION: Studies focusing on catheter removal and the pathogenic spectrum of peritoneal dialysis-associated peritonitis (PDAP) need to be updated. METHODS: Data were collected from four peritoneal dialysis (PD) centers. Peritonitis rates were compared using Poisson regression and Logistic regression was used to examine the risk factors for catheter removal. RESULTS: The PD duration (odds ratio [OR], 1.021; 95% confidence interval [CI], 1.010-1.032), number of previous PDAP episodes (OR, 1.267; 95% CI, 1.039-1.545), dialysate white cell count >100 × 106 /L on Day 5 of PDAP (OR, 6.088; 95% CI, 3.277-11.312), Pseudomonas aeruginosa (OR, 4.122; 95% CI, 1.071-15.874) and polymicrobial infections (OR, 3.257; 95% CI, 1.519-6.982) were independent predictors of catheter removal (p < 0.05). The prevalence of polymicrobial peritonitis and fungal peritonitis has increased (p < 0.05). CONCLUSION: Attention should be paid to patients with long PD duration or a history of previous episodes of PDAP characteristics.
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Diálisis Peritoneal , Peritonitis , Humanos , Estudios Retrospectivos , Diálisis Peritoneal/efectos adversos , Factores de Riesgo , Peritonitis/epidemiología , Peritonitis/etiología , Peritonitis/tratamiento farmacológico , Catéteres/efectos adversosRESUMEN
Clonal bacterial populations rely on transcriptional variation across individual cells to produce specialized states that increase fitness. Understanding all cell states requires studying isogenic bacterial populations at the single-cell level. Here we developed probe-based bacterial sequencing (ProBac-seq), a method that uses libraries of DNA probes and an existing commercial microfluidic platform to conduct bacterial single-cell RNA sequencing. We sequenced the transcriptome of thousands of individual bacterial cells per experiment, detecting several hundred transcripts per cell on average. Applied to Bacillus subtilis and Escherichia coli, ProBac-seq correctly identifies known cell states and uncovers previously unreported transcriptional heterogeneity. In the context of bacterial pathogenesis, application of the approach to Clostridium perfringens reveals heterogeneous expression of toxin by a subpopulation that can be controlled by acetate, a short-chain fatty acid highly prevalent in the gut. Overall, ProBac-seq can be used to uncover heterogeneity in isogenic microbial populations and identify perturbations that affect pathogenicity.
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Secuenciación de Nucleótidos de Alto Rendimiento , Transcriptoma , Análisis de Secuencia de ARN/métodos , Secuenciación de Nucleótidos de Alto Rendimiento/métodosRESUMEN
Neutrophil recruitment to the inflamed endothelium is a multistep process and is of utmost importance in the development of the hallmark vaso-occlusive crisis in sickle cell disease (SCD). However, there lacks a standardized, clinically feasible approach for assessing neutrophil recruitment to the inflamed endothelium for individualized risk stratification and therapeutic response prediction in SCD. Here, we describe a microfluidic device functionalized with E-selectin, a critical endothelial receptor for the neutrophil recruitment process, as a strategy to assess neutrophil binding under physiologic flow in normoxia and clinically relevant hypoxia in SCD. We show that hypoxia significantly enhances neutrophil binding to E-selectin and promotes the formation of neutrophil-platelet aggregates. Moreover, we identified two distinct patient populations: a more severe clinical phenotype with elevated lactate dehydrogenase levels and absolute reticulocyte counts but lowered fetal hemoglobin levels associated with constitutively less neutrophil binding to E-selectin. Mechanistically, we demonstrate that the extent of neutrophil activation correlates with membrane L-selectin shedding, resulting in the loss of ligand interaction sites with E-selectin. We also show that inhibition of E-selectin significantly reduces leukocyte recruitment to activated endothelial cells. Our findings add mechanistic insight into neutrophil-endothelial interactions under hypoxia and provide a clinically feasible means for assessing neutrophil binding to E-selectin using clinical whole blood samples, which can help guide therapeutic decisions for SCD patients.
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Anemia de Células Falciformes , Técnicas Biosensibles , Humanos , Selectina E/uso terapéutico , Células Endoteliales/metabolismo , Infiltración Neutrófila , Adhesión Celular , Endotelio/metabolismo , Anemia de Células Falciformes/tratamiento farmacológico , Anemia de Células Falciformes/metabolismo , Dispositivos Laboratorio en un Chip , HipoxiaRESUMEN
Rendering bridges the gap between 2D vision and 3D scenes by simulating the physical process of image formation. By inverting such renderer, one can think of a learning approach to infer 3D information from 2D images. However, standard graphics renderers involve a fundamental step called rasterization, which prevents rendering to be differentiable. Unlike the state-of-the-art differentiable renderers (Kato et al. 2018 and Loper 2018), which only approximate the rendering gradient in the backpropagation, we propose a natually differentiable rendering framework that is able to (1) directly render colorized mesh using differentiable functions and (2) back-propagate efficient supervisions to mesh vertices and their attributes from various forms of image representations. The key to our framework is a novel formulation that views rendering as an aggregation function that fuses the probabilistic contributions of all mesh triangles with respect to the rendered pixels. Such formulation enables our framework to flow gradients to the occluded and distant vertices, which cannot be achieved by the previous state-of-the-arts. We show that by using the proposed renderer, one can achieve significant improvement in 3D unsupervised single-view reconstruction both qualitatively and quantitatively. Experiments also demonstrate that our approach can handle the challenging tasks in image-based shape fitting, which remain nontrivial to existing differentiable renders.
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During the development of the vertebrate embryo, segmented structures called somites are periodically formed from the presomitic mesoderm (PSM) and give rise to the vertebral column. While somite formation has been studied in several animal models, it is less clear how well this process is conserved in humans. Recent progress has made it possible to study aspects of human paraxial mesoderm (PM) development such as the human segmentation clock in vitro using human pluripotent stem cells (hPSCs); however, somite formation has not been observed in these monolayer cultures. Here, we describe the generation of human PM organoids from hPSCs (termed Somitoids), which recapitulate the molecular, morphological, and functional features of PM development, including formation of somite-like structures in vitro. Using a quantitative image-based screen, we identify critical parameters such as initial cell number and signaling modulations that reproducibly yielded formation of somite-like structures in our organoid system. In addition, using single-cell RNA-sequencing and 3D imaging, we show that PM organoids both transcriptionally and morphologically resemble their in vivo counterparts and can be differentiated into somite derivatives. Our organoid system is reproducible and scalable, allowing for the systematic and quantitative analysis of human spine development and disease in vitro.
Humans are part of a group of animals called vertebrates, which are all the animals with backbones. Broadly, all vertebrates have a similar body shape with a head at one end and a left and right side that are similar to each other. Although this is not very obvious in humans, vertebrate bodies are derived from pairs of segments arranged from the head to the tail. Each of these segments or somites originates early in embryonic development. Cells from each somite then divide, grow and specialize to form bones such as the vertebrae of the vertebral column, muscles, skin, and other tissues that make up each segment. Studying different animals during embryonic development has provided insights into how somites form and grow, but it is technically difficult to do and only provides an approximate model of how somites develop in humans. Being able to make and study somites using human cells in the lab would help scientists learn more about how somite formation in humans is regulated. Budjan et al. grew human stem cells in the lab as three-dimensional structures called organoids, and used chemical signals similar to the ones produced in the embryo during development to make the cells form somites. Various combinations of signals were tested to find the best way to trigger somite formation. Once the somites formed, Budjan et al. measured them and studied their structure and the genes they used. They found that these lab-grown somites have the same size and structure as natural somites and use many of the same genes. This new organoid model provides a way to study human somite formation and development in the lab for the first time. This can provide insights into the development and evolution of humans and other animals that could then help scientists understand diseases such as the development of abnormal spinal curvature that affects around 1 in 10,000 newborns.
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Células Madre Pluripotentes , Somitos , Animales , Diferenciación Celular , Humanos , Mesodermo , OrganoidesRESUMEN
Acute limb ischemia (ALI) is the most severe manifestation of peripheral artery disease, accompanied by pH/temperature-microenvironment changes in two different phases. In the acute phase, temperature and pH are significantly decreased, and reactive oxygen species (ROS) are excessively generated owing to the sharp reduction of blood perfusion. Afterwards, in the chronic phase, although the temperature gradually recovers, angiogenesis is delayed due to chronic vascular injury, skeletal muscle cell apoptosis and endothelial cell dysfunction. Current therapeutic strategies mainly focus on recanalization; however, their effects on scavenging ROS in the acute phase and promoting angiogenesis in the chronic phase are quite limited. Herein, an injectable pH and temperature dual-responsive poloxamer 407 (PF127)/hydroxymethyl cellulose (HPMC)/sodium alginate (SA)-derived hydrogel (FHSgel), encapsulating melatonin and diallyl trisulfide-loaded biodegradable hollow mesoporous silica nanoparticles (DATS@dHMSNs), is developed, which can intelligently respond to the different phases of ALI. In the acute phase of ischemia, the decreased pH results in the rapid release of melatonin to scavenge excessive ischemia-induced ROS. On the other hand, in the chronic repair phase, the recovered temperature triggers the sustained release of DATS@dHMSNs from the FHSgel, thus generating hydrogen sulfide (H2S) to enhance the angiogenesis and microcirculation reconstruction of ischemic limbs.
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Sulfuro de Hidrógeno , Melatonina , Alginatos/uso terapéutico , Compuestos Alílicos , Celulosa , Preparaciones de Acción Retardada/uso terapéutico , Humanos , Hidrogeles/uso terapéutico , Sulfuro de Hidrógeno/farmacología , Concentración de Iones de Hidrógeno , Isquemia/tratamiento farmacológico , Poloxámero/uso terapéutico , Especies Reactivas de Oxígeno , Dióxido de Silicio/uso terapéutico , SulfurosRESUMEN
Background: The optimal break-in period (BI) of urgent-start peritoneal dialysis (USPD) initiation for patients with end-stage renal disease (ESRD) and diabetes is unclear. We aimed to explore the safety and applicability of a BI ≤24 h in patients with ESRD and diabetes. Methods: We used a retrospective cohort design wherein we recruited patients with ESRD and diabetes who underwent USPD at five institutions in China between January 2013 and August 2020. The enrolled patients were grouped according to BI. The primary outcomes were mechanical and infectious complication occurrences, whereas the secondary outcome was technique survival. Results: We enrolled 310 patients with diabetes, of whom 155 and 155 patients were in the BI ≤24 h and BI >24 h groups, respectively. The two groups showed a comparable incidence of infectious and mechanical complications within 6 months after catheter insertion (p>0.05). Logistic regression analysis revealed that a BI ≤24 h was not an independent risk factor for mechanical or infectious complications. Kaplan-Meier estimates showed no statistically significant between-group differences in technique survival rates (p>0.05). Cox multivariate regression analysis revealed that a BI ≤24 h was not an independent risk factor for technique failure. Conclusion: USPD initiation with a BI ≤24 h may be safe and feasible for patients with ESRD and diabetes.
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Diabetes Mellitus , Fallo Renal Crónico , Diálisis Peritoneal , Humanos , Fallo Renal Crónico/terapia , Diálisis Peritoneal/efectos adversos , Diálisis Peritoneal/métodos , Estudios Retrospectivos , Factores de TiempoRESUMEN
Thromboembolism is the leading cause of cardiovascular mortality. Currently, for the lack of targeting, short half-life, low bioavailability and high bleeding risk of the classical thrombolytic drugs, pharmacological thrombolysis is usually a slow process based on micro-pumping. In addition, frequently monitoring and regulating coagulation functions are also required during (and after) the process of thrombolysis. To address these issues, a targeted thrombolytic and anticoagulation nanoplatform (UCATS-UK) is developed based on upconversion nanoparticles (UCNPs) that can convert 808 or 980 nm near-infrared (NIR) light into UV/blue light. This nanoplatform can target and enrich in the thrombus site. Synergistic thrombolysis and anticoagulation therapy thus could be realized through the controlled release of urokinase (UK) and nitric oxide (NO). Both in vitro and in vivo experiments have confirmed the excellent thrombolytic and anticoagulative capabilities of this multifunctional nanoplatform. Combined with the unique fluorescent imaging capability of UCNPs, this work is expected to contribute to the development of clinical thrombolysis therapy towards an integrated system of imaging, diagnosis and treatment.
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The JAK2-V617F mutation is the most common cause of myeloproliferative neoplasms. Although experiments have revealed that this gain-of-function mutation is associated with myeloid blood cell expansion and increased production of white cells, red cells, and platelets, the transcriptional consequences of the JAK2-V617F mutation in different cellular compartments of the bone marrow have not yet been fully elucidated. To study the direct effects of JAK2-V617F on bone marrow cells in patients with myeloproliferative neoplasms, we performed joint single-cell RNA sequencing and JAK2 genotyping on CD34+-enriched cells from eight patients with newly diagnosed essential thrombocythemia or polycythemia vera. We found that the JAK2-V617F mutation increases the expression of interferon-response genes (e.g., HLAs) and the leptin receptor in hematopoietic progenitor cells. Furthermore, we sequenced a population of CD34- bone marrow monocytes and found that the JAK2 mutation increased expression of intermediate monocyte genes and the fibrocyte-associated surface protein SLAMF7 in these cells.
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Trastornos Mieloproliferativos , Policitemia Vera , Trombocitemia Esencial , Células de la Médula Ósea/metabolismo , Humanos , Janus Quinasa 2/genética , Janus Quinasa 2/metabolismo , Mutación , Trastornos Mieloproliferativos/genética , Policitemia Vera/genética , Trombocitemia Esencial/genéticaRESUMEN
In many biological applications, the readout of somatic mutations in individual cells is essential. For example, it can be used to mark individual cancer cells or identify progenies of a stem cell. Here, we present a protocol to perform single-cell RNA-seq and single-cell amplicon-seq using 10X Chromium technology. Our protocol demonstrates how to (1) isolate CD34+ progenitor cells from human bone marrow aspirate, (2) prepare single-cell amplicon libraries, and (3) analyze the libraries to assign somatic mutations to individual cells. For complete details on the use and execution of this protocol, please refer to Van Egeren et al. (2021).
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Mutación , Análisis de Secuencia de ARN/métodos , Análisis de la Célula Individual/métodos , Antígenos CD34 , Células de la Médula Ósea/fisiología , Cartilla de ADN , Biblioteca de Genes , HumanosRESUMEN
ABSTRACT: This study explored the effect of percutaneous intervertebral foraminoscopic discectomy (PIFD) in the treatment of lumbar disc herniation (LDH).This retrospective study collected a total of 88 patient cases for inclusion. Epidemiological and clinical data of patients with LDH at the First Affiliated Hospital of Jiamusi University between May 2017 and January 2020 were retrospectively analyzed. Of those, 44 patients received PIFD and were allocated to an intervention group. The other 44 patients administrated fenestration discectomy (FD), and were assigned to a control group. We compared surgery time (minute), incision length (cm), duration of hospital stay after surgery (day), pain intensity (as checked by Visual Analogue Scale (VAS), health-related quality of life (as examined by Oswestry Disability Index, ODI), and complications between 2 groups.There were not significant differences in surgery time (minute) (Pâ=â.56), VAS (Pâ=â.33) and ODI (Pâ=â.46) after surgery between 2 groups. However, there were significant differences in incision size (cm) (Pâ<â.01) and length of hospital stay (day) (Pâ<â.01) after surgery between 2 groups. When compared before the surgery, patients in both groups had significant improvements in VAS (Pâ<â.01) and ODI (Pâ<â.01) after the surgery. Moreover, both groups had similar safety profiles (Pâ>â.05).The findings of this study showed that both PIFD and FD benefit patients with LDH. However, PIFD can benefit patients more than FD in the incision size and duration of hospital stay after surgery.
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Discectomía Percutánea , Endoscopía , Desplazamiento del Disco Intervertebral/cirugía , Vértebras Lumbares , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Tempo Operativo , Calidad de Vida , Estudios Retrospectivos , Canal Medular , Resultado del TratamientoRESUMEN
Some cancers originate from a single mutation event in a single cell. Blood cancers known as myeloproliferative neoplasms (MPNs) are thought to originate when a driver mutation is acquired by a hematopoietic stem cell (HSC). However, when the mutation first occurs in individuals and how it affects the behavior of HSCs in their native context is not known. Here we quantified the effect of the JAK2-V617F mutation on the self-renewal and differentiation dynamics of HSCs in treatment-naive individuals with MPNs and reconstructed lineage histories of individual HSCs using somatic mutation patterns. We found that JAK2-V617F mutations occurred in a single HSC several decades before MPN diagnosis-at age 9 ± 2 years in a 34-year-old individual and at age 19 ± 3 years in a 63-year-old individual-and found that mutant HSCs have a selective advantage in both individuals. These results highlight the potential of harnessing somatic mutations to reconstruct cancer lineages.