ABSTRACT
PURPOSE: To evaluate the diagnostic performance of contrast-enhanced (CE) ultrasound using Sonazoid (SNZ-CEUS) by comparing with contrast-enhanced computed tomography (CE-CT) and contrast-enhanced magnetic resonance imaging (CE-MRI) for differentiating benign and malignant renal masses. MATERIALS AND METHODS: 306 consecutive patients (from 7 centers) with renal masses (40 benign tumors, 266 malignant tumors) diagnosed by both SNZ-CEUS, CE-CT or CE-MRI were enrolled between September 2020 and February 2021. The examinations were performed within 7 days, but the sequence was not fixed. Histologic results were available for 301 of 306 (98.37%) lesions and 5 lesions were considered benign after at least 2 year follow-up without change in size and image characteristics. The diagnostic performances were evaluated by sensitivity, specificity, positive predictive value, negative predictive value, and compared by McNemar's test. RESULTS: In the head-to-head comparison, SNZ-CEUS and CE-MRI had comparable sensitivity (95.60 vs. 94.51%, P = 0.997), specificity (65.22 vs. 73.91%, P = 0.752), positive predictive value (91.58 vs. 93.48%) and negative predictive value (78.95 vs. 77.27%); SNZ-CEUS and CE-CT showed similar sensitivity (97.31 vs. 96.24%, P = 0.724); however, SNZ-CEUS had relatively lower than specificity than CE-CT (59.09 vs. 68.18%, P = 0.683). For nodules > 4 cm, CE-MRI demonstrated higher specificity than SNZ-CEUS (90.91 vs. 72.73%, P = 0.617) without compromise the sensitivity. CONCLUSIONS: SNZ-CEUS, CE-CT, and CE-MRI demonstrate desirable and comparable sensitivity for the differentiation of renal mass. However, the specificity of all three imaging modalities is not satisfactory. SNZ-CEUS may be a suitable alternative modality for patients with renal dysfunction and those allergic to gadolinium or iodine-based agents.
Subject(s)
Contrast Media , Ferric Compounds , Iron , Kidney Neoplasms , Magnetic Resonance Imaging , Oxides , Tomography, X-Ray Computed , Ultrasonography , Humans , Kidney Neoplasms/diagnostic imaging , Kidney Neoplasms/pathology , Male , Female , Middle Aged , Prospective Studies , Ultrasonography/methods , Tomography, X-Ray Computed/methods , Magnetic Resonance Imaging/methods , Aged , Diagnosis, Differential , Adult , Aged, 80 and overABSTRACT
BACKGROUND: TAP1 is an immunomodulation-related protein that plays different roles in various malignancies. This study investigated the transcriptional expression profile of TAP1 in uveal melanoma (UVM), revealed its potential biological interaction network, and determined its prognostic value. METHODS: CIBERSORT and ESTIMATE bioinformatic methods were used on data sourced from The Cancer Genome Atlas database (TCGA) to determine the correlation between TAP1 expression, UVM prognosis, biological characteristics, and immune infiltration. Gene set enrichment analysis (GSEA) was used to discover the signaling pathways associated with TAP1, while STRING database and CytoHubba were used to construct protein-protein interaction (PPI) and competing endogenous RNA (ceRNA) networks, respectively. An overall survival (OS) prognostic model was constructed to test the predictive efficacy of TAP1, and its effect on the in vitro proliferation activity and metastatic potential of UVM cell line C918 cells was verified by RNA interference. RESULTS: There was a clear association between TAP1 expression and UVM patient prognosis. Upregulated TAP1 was strongly associated with a shorter survival time, higher likelihood of metastasis, and higher mortality outcomes. According to GSEA analysis, various immunity-related signaling pathways such as primary immunodeficiency were enriched in the presence of elevated TAP1 expression. A PPI network and a ceRNA network were constructed to show the interactions among mRNAs, miRNAs, and lncRNAs. Furthermore, TAP1 expression showed a significant positive correlation with immunoscore, stromal score, CD8+ T cells, and dendritic cells, whereas the correlation with B cells and neutrophils was negative. The Cox regression model and calibration plots confirmed a strong agreement between the estimated OS and actual observed patient values. In vitro silencing of TAP1 expression in C918 cells significantly inhibited cell proliferation and metastasis. CONCLUSIONS: This study is the first to demonstrate that TAP1 expression is positively correlated with clinicopathological factors and poor prognosis in UVM. In vitro experiments also verified that TAP1 is associated with C918 cell proliferation, apoptosis, and metastasis. These results suggest that TAP1 may function as an oncogene, prognostic marker, and importantly, as a novel therapeutic target in patients with UVM.
Subject(s)
Biomarkers, Tumor , MicroRNAs , Humans , Biomarkers, Tumor/genetics , Gene Regulatory Networks , MicroRNAs/genetics , MicroRNAs/metabolism , Prognosis , ATP Binding Cassette Transporter, Subfamily B, Member 2/geneticsABSTRACT
Prostaglandin F2α (PGF2α), the first-line anti-glaucoma medication, can cause the deepening of the upper eyelid sulcus due to orbital lipoatrophy. However, the pathogenesis of Graves' ophthalmopathy (GO) involves the excessive adipogenesis of the orbital tissues. The present study aimed to determine the therapeutic effects and underlying mechanisms of PGF2α on adipocyte differentiation. In this study primary cultures of orbital fibroblasts (OFs) from six patients with GO were established. Immunohistochemistry, immunofluorescence, and Western blotting (WB) were used to evaluated the expression of the F-prostanoid receptor (FPR) in the orbital adipose tissues and the OFs of GO patients. The OFs were induced to differentiate into adipocytes and treated with different incubation times and concentrations of PGF2α. The results of Oil red O staining showed that the number and size of the lipid droplets decreased with increasing concentrations of PGF2α and the reverse transcription-polymerase chain reaction (RT-PCR) and WB of the peroxisome proliferator-activated receptor γ (PPARγ) and fatty-acid-binding protein 4 (FABP4), both adipogenic markers, were significantly downregulated via PGF2α treatment. Additionally, we found the adipogenesis induction of OFs promoted ERK phosphorylation, whereas PGF2α further induced ERK phosphorylation. We used Ebopiprant (FPR antagonist) to interfere with PGF2α binding to the FPR and U0126, an Extracellular Signal-Regulated Kinase (ERK) inhibitor, to inhibit ERK phosphorylation. The results of Oil red O staining and expression of adipogenic markers showed that blocking the receptor binding or decreasing the phosphorylation state of the ERK both alleviate the inhibitory effect of PGF2a on the OFs adipogenesis. Overall, PGF2α mediated the inhibitory effect of the OFs adipogenesis through the hyperactivation of ERK phosphorylation via coupling with the FPR. Our study provides a further theoretical reference for the potential application of PGF2α in patients with GO.
Subject(s)
Dinoprost , Graves Ophthalmopathy , Humans , Dinoprost/metabolism , Adipogenesis , Extracellular Signal-Regulated MAP Kinases/metabolism , Graves Ophthalmopathy/pathology , Fibroblasts/metabolism , Cells, CulturedABSTRACT
Pancreatic cancer is a highly lethal cancer characterized by local invasiveness, early metastasis, recurrence and high resistance to current therapies. Extensive stroma or desmoplasia is a key histological feature of the disease, and interactions between cancer and stromal cells are critical for pancreatic cancer development and progression. Mesenchymal stromal cells [MSCs] exhibit preferential tropism to primary and metastatic tumor sites and may either suppress or support tumor growth. Although MSCs represent a potential source of pancreatic cancer stroma, their contribution to pancreatic tumor growth remains poorly known. Here, we show that bone marrow MSCs significantly contribute to pancreatic cancer growth in vitro and in vivo. Furthermore, MSCs create a pro-carcinogenic microenvironment through the release of key factors mediating growth and angiogenesis, including interleukin (IL)-6, IL-8, vascular endothelial growth factor and activation of STAT3 signaling in tumor cells. IL-6 released by MSCs was largely responsible for the pro-tumorigenic effects of MSCs. Knockdown of IL-6 expression in MSCs by small interfering RNA (siRNA) abolished the MSC growth-promoting effect in vitro, reducing tumor cell proliferation and clonogenic potential. In addition, in a heterotopic nude mouse model of human pancreatic tumor xenografts, blockade of IL-6 with the anti-IL-6 receptor antibody, tocilizumab, or of its downstream effector STAT3 with the small molecule STAT3 inhibitor S3I-201, abrogated MSC-mediated tumor promotion and delayed tumor formation significantly. Our data demonstrate that MSCs promote pancreatic cancer growth, with IL-6 produced by MSCs playing a pivotal role.
Subject(s)
Interleukin-6/metabolism , Mesenchymal Stem Cells , Pancreatic Neoplasms , Animals , Bone Marrow/metabolism , Cell Line, Tumor , Humans , Mice , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/therapy , Tumor Microenvironment , Vascular Endothelial Growth Factor A/metabolism , Pancreatic NeoplasmsABSTRACT
An aerobic, Gram-staining-positive, rod-shaped, endospore-forming and motile bacterial strain, designated SJY2T, was isolated from the rhizosphere soil of tea plants (Camellia sinensis var. assamica) collected in the organic tea garden of the Jingmai Pu-erh tea district in Pu'er city, Yunnan, southwest China. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolate belonged to the genus Paenibacillus. The closest phylogenetic relative was Paenibacillus filicis DSM 23916T (98.1% similarity). The major fatty acids (> 10% of the total fatty acids) were anteiso-C15:0 and isoC16:0. The major respiratory quinone was MK-7 and the major polar lipid was diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylmonomethylethanolamine. The peptidoglycan contained glutamic acid, serine, alanine and meso-diaminopimelic acid. Genome sequencing revealed a genome size of 6.71 Mbp and a G + C content of 53.1%. Pairwise determined whole genome average nucleotide identity (gANI) values and digital DNA-DNA hybridization (dDDH) values suggested that strain SJY2T represents a new species, for which we propose the name Paenibacillus puerhi sp. nov. with the type strain SJY2T (= CGMCC 1.17156T = KCTC 43242T).
Subject(s)
Camellia sinensis/microbiology , Paenibacillus/classification , Rhizosphere , Soil Microbiology , Benzoquinones/analysis , China , DNA, Bacterial/genetics , Fatty Acids/analysis , Genome, Bacterial/genetics , Paenibacillus/chemistry , Paenibacillus/genetics , Paenibacillus/physiology , Peptidoglycan/analysis , Phospholipids/analysis , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
During oocyte meiosis, mitochondria usually surround spindle to meet the energy demand of spindle migration and chromosome segregation. Therefore, the mitochondrion surrounding spindle is widely accepted as an important indicator to demonstrate the mitochondrial function in oocyte studies. However, the role of mitochondria surrounding spindle in oocyte quality is not exactly addressed. Mitofusin-2 (MFN2) is a mitochondrial outer membrane GTPase that mediates mitochondrial clustering and fusion. Here, we increased the mitochondria surrounding spindle by overexpression of MFN2 in mouse oocytes. Results indicate that the increase of mitochondria surrounding spindle has little effect on germinal vesicle breakdown (GVBD), spindle migration, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) production and Endoplasmic reticulum (ER) distribution, while blocks chromosome segregation, destroys the spindle, and finally causes most of the oocytes to arrest at metaphase I stage. Collectively, our results demonstrate the mitochondria surrounding spindle is precisely regulated during oocyte maturation, while too much of it may cause abnormal oocyte meiosis. Therefore, although mitochondrion surrounding spindle is a typical biological event during oocyte maturation, utilizing it to demonstrate the mitochondrial function and oocyte quality should be much careful.
Subject(s)
Metaphase , Mitochondria/metabolism , Oocytes/cytology , Spindle Apparatus/metabolism , Animals , Cells, Cultured , Female , GTP Phosphohydrolases/genetics , GTP Phosphohydrolases/metabolism , HeLa Cells , Humans , Membrane Potential, Mitochondrial , Mice , Mice, Inbred ICR , Mitochondria/genetics , Oocytes/metabolism , Oogenesis , Reactive Oxygen Species/metabolism , Spindle Apparatus/genetics , Up-RegulationABSTRACT
Mesenchymal stromal cells (MSCs) are the subject of numerous clinical trials, largely due to their immunomodulatory and tissue regenerative properties. Toll-like receptors (TLRs), especially TLR3 and TLR4, are highly expressed on MSCs and their activation can significantly modulate the immunosuppressive and anti-inflammatory functions of MSCs. While MSCs can recruit and promote the generation of regulatory T cells (Tregs), the effect of TLR activation on MSC-mediated Treg induction is unknown. In this study, we investigated the effect of ligand-mediated activation of TLR3 and TLR4 on Treg induction by human MSCs. We found that generation of Tregs in human CD4(+) lymphocyte and MSC cocultures was enhanced by either TLR3 or TLR4 activation of MSCs and that the increase was abolished by TLR3 and TLR4 gene-silencing. Augmented Treg induction by TLR-activated MSCs was cell contact-dependent and associated with increased gene expression of the Notch ligand, Delta-like 1. Moreover, inhibition of Notch signaling abrogated the augmented Treg levels in the MSC cocultures. Our data show that TLR3 or TLR4 activation of MSCs increases Treg induction via the Notch pathway and suggest new means to enhance the potency of MSCs for treating disorders with an underlying immune dysfunction, including steroid resistant acute graft-versus-host disease. Stem Cells 2017;35:265-275.
Subject(s)
Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Receptors, Notch/metabolism , Signal Transduction , Toll-Like Receptor 3/metabolism , Toll-Like Receptor 4/metabolism , Antigens, CD/metabolism , Cell Communication , Gene Silencing , Humans , Immunosuppression Therapy , Ligands , T-Lymphocytes, Regulatory , Transplantation, HomologousABSTRACT
Patients with acute myeloid leukemia (AML) often relapse after initial therapy because of persistence of leukemic stem cells that frequently express the IL-3 receptor alpha chain CD123. Natural killer (NK) cell-based therapeutic strategies for AML show promise and we explore the NK cell lines, NK-92 and CD16+ NK-92, as a treatment for AML. NK-92 has been tested in phase I clinical trials with minimal toxicity; irradiation prior to infusion prevents risk of engraftment. The CD16 negative NK-92 parental line was genetically modified to express the high affinity Fc gamma receptor, enabling antibody-dependent cell-mediated cytotoxicity, which we utilized in combination with an anti-CD123 antibody to target leukemic stem cells. NK-92 was preferentially cytotoxic against leukemic stem and progenitor cells compared with bulk leukemia in in vitro assays, while CD16+ NK-92 in combination with an anti-CD123 mAb mediated antibody-dependent cell-mediated cytotoxicity against CD123+ leukemic targets. Furthermore, NK-92 infusions (with or without prior irradiation) improved survival in a primary AML xenograft model. Mice xenografted with primary human AML cells had a superior survival when treated with irradiated CD16+NK-92 cells and an anti-CD123 monoclonal antibody (7G3) versus treatment with irradiated CD16+NK-92 cells combined with an isotype control antibody. In this proof-of-principle study, we show for the first time that a CD16+NK-92 cell line combined with an antibody that targets a leukemic stem cell antigen can lead to improved survival in a relevant pre-clinical model of AML.
Subject(s)
Antineoplastic Agents, Immunological/pharmacology , Interleukin-3 Receptor alpha Subunit/antagonists & inhibitors , Leukemia, Myeloid, Acute/drug therapy , Neoplasm Proteins/antagonists & inhibitors , Receptors, IgG/antagonists & inhibitors , Animals , GPI-Linked Proteins/antagonists & inhibitors , GPI-Linked Proteins/metabolism , Humans , Interleukin-3 Receptor alpha Subunit/metabolism , K562 Cells , Leukemia, Myeloid, Acute/metabolism , Leukemia, Myeloid, Acute/pathology , Mice , Mice, Inbred NOD , Mice, SCID , Neoplasm Proteins/metabolism , Receptors, IgG/metabolism , Xenograft Model Antitumor AssaysABSTRACT
BACKGROUND: Mesenchymal stromal cells (MSCs) promote wound healing, including after radiotherapy (RT) and surgery. The use of MSCs in regenerative medicine in the context of malignancy, such as to enhance wound healing post-RT/surgery in patients with soft tissue sarcomas (STSs), requires safety validation. The aim of this study was to determine the effects of human MSCs on STS growth in vitro and local recurrence and metastasis in vivo. METHODS: Human primary STS and HT-1080 fibrosarcoma lines were transduced to express luciferase/eGFP (enhanced green fluorescent protein). Sarcoma cells were co-cultured or co-injected with bone marrow-derived MSCs for growth studies. Xenograft tumor models were established with STS lines in NOD/SCID/γcnull mice. To emulate a clinical scenario, subcutaneous tumors were treated with RT/surgery prior to MSC injection into the tumor bed. Local and distant tumor recurrence was studied using histology and bioluminescence imaging. RESULTS: MSCs did not promote STS proliferation upon co-culture in vitro, which was consistent among MSCs from different donors. Co-injection of MSCs with sarcoma cells in mice exhibited no significant tumor-stimulating effect, compared with control mice injected with sarcoma cells alone. MSC administration after RT/surgery had no effect on local recurrence or metastasis of STS. DISCUSSION: These studies are important for the establishment of a safety profile for MSC administration in patients with STS. Our data suggest that MSCs are safe in STS management after standard of care RT/surgery, which can be further investigated in early-phase clinical trials to also determine the efficacy of MSCs in reducing morbidity and to mitigate wound complications in these patients.
Subject(s)
Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells/physiology , Radiotherapy , Sarcoma/pathology , Sarcoma/therapy , Surgical Procedures, Operative , Adult , Animals , Coculture Techniques , Combined Modality Therapy , HEK293 Cells , Heterografts , Humans , Male , Mesenchymal Stem Cell Transplantation/adverse effects , Mice , Mice, Inbred NOD , Mice, SCID , Mice, Transgenic , Neoplasm Recurrence, Local/etiology , Neoplasm Recurrence, Local/pathology , Radiotherapy/adverse effects , Radiotherapy/methods , Surgical Procedures, Operative/adverse effects , Surgical Procedures, Operative/methods , Tumor Cells, Cultured , Wound Healing , Xenograft Model Antitumor AssaysABSTRACT
BACKGROUND: The MADS-box transcription factor myocyte enhancer factor 2C (MEF2C) is required for the cardiac development and postnatal adaptation and in mice-targeted disruption of the MEF2C gene results in dilated cardiomyopathy (DCM). However, in humans, the association of MEF2C variation with DCM remains to be investigated. METHODS: The coding regions and splicing boundaries of the MEF2C gene were sequenced in 172 unrelated patients with idiopathic DCM. The available close relatives of the index patient harboring an identified MEF2C mutation and 300 unrelated, ethnically matched healthy individuals used as controls were genotyped for MEF2C. The functional effect of the mutant MEF2C protein was characterized in contrast to its wild-type counterpart by using a dual-luciferase reporter assay system. RESULTS: A novel heterozygous MEF2C mutation, p.Y157X, was detected in an index patient with adult-onset DCM. Genetic screen of the mutation carrier's family members revealed that the mutation co-segregated with DCM, which was transmitted as an autosomal dominant trait with complete penetrance. The non-sense mutation was absent in 300 control individuals. Functional analyses unveiled that the mutant MEF2C protein had no transcriptional activity. Furthermore, the mutation abolished the synergistic transactivation between MEF2C and GATA4 as well as HAND1, two other transcription factors that have been associated with DCM. CONCLUSIONS: This study indicates MEF2C as a new gene responsible for human DCM, which provides novel insight into the mechanism underpinning DCM, suggesting potential implications for development of innovative prophylactic and therapeutic strategies for DCM, the most prevalent form of primary myocardial disease.
Subject(s)
Cardiomyopathy, Dilated/genetics , Adult , Cardiomyopathy, Dilated/metabolism , Female , HeLa Cells , Humans , MEF2 Transcription Factors/deficiency , MEF2 Transcription Factors/genetics , MEF2 Transcription Factors/metabolism , Male , Middle Aged , Mutation , Tumor Cells, CulturedABSTRACT
To improve the sensitivity of the molecular imprinting sensor detection of protein, a new strategy based on enzyme amplification was proposed. The determination of bovine serum albumin (BSA) was achieved by using the epitope imprinted techniques coupling with electrochemical measurement method. Nonapeptide, separated from BSA, was selected as a template molecule to prepare the molecularly imprinted polymer (MIP) film, and it could bind with the cavities of the MIP. By the use of epitope imprinted techniques, BSA can be recognized by the MIP via the nonapeptide on the surface of BSA. The synthesized horseradish peroxidase-labeled nonapeptide (HRP-nonapeptide) can also be recognized by the MIP. After the competitive reaction between HRP-nonapeptide and BSA, the enzymatic reaction derived from labeled HRP on the H2O2-hydroquinone system make the electrochemical current of hydroquinone change, then the concentration of BSA can be indirectly determined. BSA in the range of 1.0-150 ng/mL exhibited a linear relationship with the differential pulse voltammetric current variation and the detection limit was 0.02 ng/mL. The sensor has high sensitivity, good selectivity, and reproducibility. It has been applied to the determination of residual bovine serum albumin in human rabies vaccine with the recovery rate of 98.3%-102.5%.
Subject(s)
Biosensing Techniques/methods , Electrochemical Techniques/methods , Molecular Imprinting/methods , Peptide Fragments/chemistry , Polymers/chemistry , Rabies Vaccines/analysis , Serum Albumin, Bovine/analysis , Animals , Cattle , Chlorocebus aethiops , Electrodes , Horseradish Peroxidase/chemistry , Horseradish Peroxidase/metabolism , Humans , Hydrogen Peroxide/chemistry , Limit of Detection , Reproducibility of Results , Serum Albumin, Bovine/chemistry , Serum Albumin, Bovine/isolation & purification , Spectroscopy, Fourier Transform Infrared , Vero CellsABSTRACT
Objective To explore the ultrasound features and levels of cervical lymph node metastases in primary and recurrent/persistent papillary thyroid cancer (PTC).Methods We retrospectively analyzed the clinical data of 2181 patients who underwent cervical lymph nodes dissection for PTC from January 1st 2015 to January 1st 2016.Totally 418 PTC patients (with 622 lymph nodes) who met the inclusion criteria entered the final analysis.Patients who had not received any prior thyroid treatment (surgery with or without radioactive iodine) were categorized as the primary group (352 patients with 527 metastatic lymph nodes),and patients who had received prior treatment (thyroidectomy with or without radioactive iodine) for PTC were categorized as recurrent/persistent group (66 patients with 95 metastatic lymph nodes).Pathological results from lymph node dissections were used as the gold standards by means of level-to-level analysis.Results The mean of the minimum axis diameter of the lymph nodes in the primary group was (6.7±3.6)mm,and that of the recurrent/persistent group was (6.6±3.1)mm (U=0.180,P=0.857).The proportion of metastasis in the central area of primary group was 40.0%,which was significantly higher than that in the recurrent/persistent group (12.6%);the proportion of metastasis in the lateral area was 60.6% in the primary group,which was significantly lower than that in the recurrent/persistent group (87.4%)(χ2=26.288,P<0.001).In lateral metastatic lymph nodes,â ¢ level was the most common place in both groups.Level â ¤ metastatic lymph was rare in both primary group and recurrent/persistent group.Calcifications (63.1% vs. 48.2%;χ2=7.207,P=0.007) and peripheral vascularity (81.1% vs. 59.4%;χ2= 16.147, P<0.001) were more common in the recurrent/persistent group.The round shape,absence of an echogenic hilum,hyperechogenicity,and cystic aspects were not significantly different between these two groups (all P>0.05).Conclusions Primary metastatic lymph nodes often occur in the central area of lymph nodes,while lateral metastatic lymph nodes are more common in recurrent/persistent PTC.For metastatic lymph nodes,calcifications and peripheral vascularity are more common in recurrent/persistent PTC.
Subject(s)
Carcinoma, Papillary/diagnostic imaging , Lymph Nodes/diagnostic imaging , Thyroid Cancer, Papillary/diagnostic imaging , Thyroid Neoplasms/diagnostic imaging , Humans , Lymphatic Metastasis/diagnostic imaging , Neoplasm Recurrence, Local/diagnostic imaging , Retrospective Studies , ThyroidectomyABSTRACT
BACKGROUND AIMS: The use of bone marrow-derived mesenchymal stromal cells (MSCs) in cell-based therapies is currently being developed for a number of diseases. Thus far, the clinical results have been inconclusive and variable, in part because of the variety of cell isolation procedures and culture conditions used in each study. A new isolation technique that streamlines the method of concentration and demands less time and attention could provide clinical and economic advantages compared with current methodologies. In this study, we evaluated the concentrating capability of an integrated centrifuge-based technology compared with standard Ficoll isolation. METHODS: MSCs were concentrated from bone marrow aspirate using the new device and the Ficoll method. The isolation capabilities of the device and the growth characteristics, secretome production, and differentiation capacity of the derived cells were determined. RESULTS: The new MSC isolation device concentrated the bone marrow in 90 seconds and resulted in a mononuclear cell yield 10-fold higher and with a twofold increase in cell retention compared with Ficoll. The cells isolated using the device were shown to exhibit similar morphology and functional activity as assessed by growth curves and secretome production compared to the Ficoll-isolated cells. The surface marker and trilineage differentiation profile of the device-isolated cells was consistent with the known profile of MSCs. DISCUSSION: The faster time to isolation and greater cell yield of the integrated centrifuge-based technology may make this an improved approach for MSC isolation from bone marrow aspirates.
Subject(s)
Bone Marrow Cells/cytology , Cell Separation/methods , Centrifugation/methods , Mesenchymal Stem Cells/cytology , Bone Marrow , Cell Differentiation/physiology , Cell Proliferation , Cell- and Tissue-Based Therapy/methods , Ficoll , Humans , Osteoblasts/cytologyABSTRACT
This study deciphered the molecular mechanisms of the inhibition of MMP-9 expression using rosuvastatin in cultured human umbilical vein endothelial cells (HUVECs) and apoE knockout mice and whether the combination of rosuvastatin and probucol enhanced this effect. The role that microRNA (miR)-497 plays in the regulation of MMP-9 expression was evaluated in cultured HUVECs and apoE knockout mice using quantitative real-time reverse transcription polymerase chain reaction and Western blotting. First, TNFα significantly increased mitogen-activated protein kinases/extracellular signal-regulated kinase (MAPK/ERK) signaling and MMP-9 levels, and the transfection of miR-497 prevented this increase. The converse results were obtained after miR-497 suppression. Second, the administration of rosuvastatin or the combination of two drugs decreased MAPK/ERK signaling and MMP-9 levels, and the suppression of miR-497 upregulated these levels. Third, the administration of rosuvastatin or the combination of two drugs increased miR-497 expression levels in the aortas of apoE knockout mice, but the levels of serum lipids and plaque areas decreased, which improved plaque components and decreased the MAPK/ERK signaling and MMP-9 levels. Finally, the combination of the two drugs was more effective than the use of rosuvastatin alone. Rosuvastatin inhibits MMP-9 expression by upregulating miR-497 in HUVECs and apoE knockout mice, and the combination of rosuvastatin and probucol enhances this effect.
Subject(s)
Apolipoproteins E/deficiency , Gene Expression Regulation, Enzymologic/drug effects , Human Umbilical Vein Endothelial Cells/metabolism , Matrix Metalloproteinase 9/biosynthesis , MicroRNAs/biosynthesis , Probucol/pharmacology , Rosuvastatin Calcium/pharmacology , Animals , Humans , Matrix Metalloproteinase 9/genetics , Mice , Mice, Knockout , MicroRNAs/geneticsABSTRACT
OBJECTIVES: To investigate the effect of inflammation on fibrosis staging measured by quantitative elasticity parameters in rats with immune hepatitis. METHODS: Fifty-two rats were injected with swine serum as a model group, whereas 8 rats were injected with saline as a control group. Rats were randomly subjected to real-time tissue elastography biweekly. Tissue dispersion quantitative analysis was performed to obtain 12 quantitative elasticity parameters: relative mean value, standard deviation, blue area percentage, complexity, kurtosis, skewness, contrast, entropy, inverse difference moment, angular second moment, correlation, and liver fibrosis index. Subsequently, rats were euthanized, and liver specimens were taken for pathologic examination. The Kruskal-Wallis test was used for comparisons among groups. Spearman rank correlation analysis was used for correlation analysis. Receiver operating characteristic curves were used to optimize cutoff values and evaluate the diagnostic performance of the liver fibrosis index. RESULTS: Except for complexity, kurtosis, and correlation, the other 9 parameters had statistical differences (P < .05), and among these 9 parameters, the liver fibrosis index had the strongest correlation with fibrosis staging (r = 0.809; P < .05). Except for kurtosis and correlation, the other 10 parameters had statistical differences (P < .05), and among these 10 parameters, the liver fibrosis index had the highest correlation with inflammation grading (r= 0.766; P< .05). The fibrosis index cutoff values were 2.35 for stage S1 or higher (area under the curve [AUC], 0.940; sensitivity, 97.1%; specificity, 73.3%), 2.99 for stage S2 or higher (AUC, 0.865; sensitivity, 78.6%; specificity, 81.0%), 3.48 for stage S3 or higher (AUC, 0.924; sensitivity, 94.4%; specificity, 87.1%), and 4.05 for stage S4 (AUC, 0.933; sensitivity, 87.5%; specificity, 95.1%). CONCLUSIONS: Real-time elastography could be used to noninvasively evaluate fibrosis staging in rats with immune hepatitis. However, inflammation had an effect on the accuracy of this technique.
Subject(s)
Elasticity Imaging Techniques/methods , Hepatitis, Autoimmune/complications , Inflammation/complications , Liver Cirrhosis/complications , Liver Cirrhosis/diagnostic imaging , Animals , Area Under Curve , Disease Models, Animal , Inflammation/diagnostic imaging , Liver/diagnostic imaging , Liver/pathology , Liver Cirrhosis/pathology , Male , Rats , Rats, Sprague-Dawley , Reproducibility of Results , Sensitivity and Specificity , Severity of Illness Index , SwineABSTRACT
BACKGROUND: Shenghui soup is a traditional Chinese herbal medicine used in clinic for the treatment of forgetfulness. In order to understanding the prescription principle, the effects of "tonifying qi and strengthening spleen" group (TQSS) including Poria cocos (Schw.) Wolf. and Panax ginseng C.A.Mey and "eliminating phlegm and strengthening intelligence" group (EPSI) composed of Polygala tenuifolia Willd., Acorus calamus L. and Sinapis alba L from the herb complex on neurite growth in PC12 cells, two disassembled prescriptions derived from Shenghui soup and their molecular mechanisms were investigated. METHODS: Firstly, CCK-8 kit was used to detect the impact of the two prescriptions on PC12 cell viability; and Flow cytometry was performed to measure the cell apoptosis when PC12 cells were treated with these drugs. Secondly, the effect of the two prescriptions on the differentiation of PC12 cells was observed. Finally, the mRNA and protein expression levels of GAP-43 were analyzed by RT-PCR and western blot, respectively. RESULTS: "Tonifying qi and strengthening spleen" prescription decreased cell viability in a dose-dependent manner, but had no significant effect on cell apoptosis. Meanwhile, it could improve neurite growth and elevate the mRNA and protein expression level of GAP-43. "Eliminating phlegm and strengthening intelligence" prescription also exerted the similar effects on cell viability and apoptosis. Furthermore, it could also enhance cell neurite growth, with a higher expression level of GAP-43 mRNA and protein. CONCLUSION: "Tonifying qi and strengthening spleen" and "eliminating phlegm and strengthening intelligence" prescriptions from Shenghui soup have a positive effect on neurite growth. Their effects are related to the up-regulating expression of GAP-43.
Subject(s)
Drugs, Chinese Herbal/pharmacology , GAP-43 Protein/metabolism , Gene Expression/drug effects , Neurites/drug effects , Animals , GAP-43 Protein/genetics , PC12 Cells , RNA, Messenger/genetics , RNA, Messenger/metabolism , RatsABSTRACT
The inbred rat is a suitable model for studying human disease and because of its larger size is more amenable to complex surgical manipulation than the mouse. While the rodent fulfills many of the criteria for transplantation research, an important requirement is the ability to mark and track donors cells and assess organ viability. However, tracking ability is limited by the availability of transgenic (Tg) rats that express suitable luminescent or fluorescent proteins. Red fluorescent protein cloned from Discosoma coral (DsRed) has several advantages over other fluorescent proteins, including in vivo detection in the whole animal and ex vivo visualization in organs as there is no interference with autofluorescence. We generated and characterized a novel inbred Tg Lewis rat strain expressing DsRed monomeric (DsRed mono) fluorescent protein under the control of a ubiquitously expressed ROSA26 promoter. DsRed mono Tg rats ubiquitously expressed the marker gene as detected by RT-PCR but the protein was expressed at varying levels in different organs. Conventional skin grafting experiments showed acceptance of DsRed monomeric Tg rat skin on wild-type rats for more than 30 days. Cardiac transplantation of DsRed monomeric Tg rat hearts into wild-type recipients further showed graft acceptance and long-term organ viability (>6 months). The DsRed monomeric Tg rat provides marked cells and/or organs that can be followed for long periods without immune rejection and therefore is a suitable model to investigate cell tracking and organ transplantation.
Subject(s)
Animals, Genetically Modified/genetics , Luminescent Proteins/genetics , Rats, Inbred Strains/genetics , Animals , Animals, Genetically Modified/metabolism , Heart Transplantation/methods , Luminescent Proteins/immunology , Luminescent Proteins/metabolism , Magnetic Resonance Imaging , Rats , Reverse Transcriptase Polymerase Chain Reaction , Skin Transplantation/methodsABSTRACT
Surface glossiness is one of the important visual appearance parameters of natural polymer material (wood) and its related products. To realize the fast measurement of natural polymer material surface glossiness is of great significance to the online quality control and assessment of its surface. In order to broaden the application of near infrared (NIR) spectroscopy in the field of polymer material surface quality control and realize the feasibility of NIR as a fast measurement of surface glossiness, the NIR combined with partial least squares (PLS) analysis were used to analyse the correlations of natural polymer material wood surface glossiness between the NIR predicted and lab measured, and then to investigate the feasibility of NIR to rapidly predict the surface glossiness of natural polymer material wood. The results showed that the wood NIR diffuse reflectance spectroscopy regularly varied with the different wood surface glossiness, from which we can concluded that the NIR spectrums reflected the information of wood surface glossiness. The correlation coefficients of surface glossiness between the PLS models predicted and lab measured were up to 0.90. Additionally, by changing the degree between the fiber and sample surface, we collected the different wood NIR spectrums, the accuracy of NIR surface glossiness models based on these NIR spectrums had not significantly improved, and models based on the NIR spectrums collected by the 90 degree between the fiber and sample surface performed better.
Subject(s)
Spectroscopy, Near-Infrared , Wood/analysis , Least-Squares Analysis , Polymers , Quality Control , Surface PropertiesABSTRACT
BACKGROUND: Cerebral palsy (CP) is characterized by abnormal pronunciation, posture, and movement. Spastic CP accounts for more than 70% of all CP. To date, there has been no bibliometric analysis to summarize study on spastic CP. Here, we aim to conduct a bibliometric analysis of spastic CP to summarize this field's knowledge structure, research hotspots, and frontiers. METHOD: Publications about spastic CP were searched utilizing the Web of Science Core Collection (WoSCC) database from 1 January 2000 to 30 November 2022, the WoSCC literature analysis wire, VOSviewer 1.6.18, CiteSpace 6.1.R4 and Online analysis platform for bibliometrics were used to conduct the analysis. RESULTS: A total of 3988 publications, consisting of 3699 articles and 289 reviews, were included in our study. The United States emerged as the most productive country, while Kathleen Univ Leuven was the most productive institution. The leading author was Desloovere K. A total of 238 journals contributed to this field, with Developmental medicine and child neurology being the leading journal. Important keywords and keyword clusters included Spastic cerebral palsy, Reliability, and Gross motor function. Keywords identified through burst detection indicated that hotspots in this field were management, randomized controlled trials, and definition. CONCLUSION: Based on the analysis of bibliometric on spastic CP over the past 20 years, the trends and the knowledge graph of the countries, institutions, authors, references, and the keywords have been identified, providing accurate and expedited insights into critical information and potentially new directions in the study of spastic CP.