JoVE Visualize What is visualize?
Stop Reading. Start Watching.
Advanced Search
Stop Reading. Start Watching.
Regular Search
Find video protocols related to scientific articles indexed in Pubmed.
The dose-effect relationship between the seeding quantity of human marrow mesenchymal stem cells and in vivo tissue engineered bone yield.
Cell Transplant
PUBLISHED: 11-15-2014
Show Abstract
Hide Abstract
Although the feasibility of human bone marrow mesenchymal stem cells (hBMMSCs) based tissue engineered bone (TEB) has been proven in a number of studies, reaching a high positive fraction and bone yield of TEB still remains a challenge. Here, we report a dose-effect relationship of the quantity of seeded cell with in vivo bone yield, and the required quantity of hBMMSCs for the effective, stable bone formation of TEB. In our study, TEB was constructed using the static seeding technique with the gradient of seeding densities and volumes of Passage 3 hBMMSCs. The in vitro characteristics of seeding efficiency, proliferation, viability, distribution, and osteogenic differentiation of hBMMSCs seeded on two commercial scaffolds of ?-TCP and CHA were investigated using alamar blue assay, live/dead staining, confocal laser scanning microscope (CLSM), scanning electronic microscopy (SEM) examination, and mRNA expression analysis of osteogenic differentiation markers. After 3 months of ectopic implantation, in vivo bone regeneration was examined by quantitative analysis of histology and Micro-CT. The results showed that 10×10(6) cells/mL was the minimum cell seeding density for CHA and ?-TCP to generate new bone in vivo. In addition, 20×10(6) cells/mL and 30×10(6) cells/mL were the saturating seeding densities for CHA and ?-TCP to produce new bone effectively and stably. Thus for different scaffold, the saturating seeding density should be investigated firstly to ensure the effectiveness and stability of TEB construction with minimum donor injury, which is essential for the clinical application of TEB.
Related JoVE Video
Pirenzepine Inhibits Myopia in Guinea Pig Model by Regulating the Balance of MMP-2 and TIMP-2 Expression and Increased Tyrosine Hydroxylase Levels.
Cell Biochem. Biophys.
PUBLISHED: 11-13-2014
Show Abstract
Hide Abstract
In this study, we investigated the effects and mechanism of action of pirenzepine in a guinea pig model of myopia induced by exposure to monochromatic light. It was observed that pirenzepine inhibited the increase of diopter and extension of ocular axial length. Immunohistochemistry staining showed that the number of tyrosine hydroxylase (TH)-positive cells in pirenzepine group was significantly higher compared to the other treatment groups pointing to a highly positive correlation between TH expression levels and the diopter and axial length change. RT-PCR analysis further showed that pirenzepine treatment reduced the expression of matrix metalloproteinase (MMP-2) and enhanced the expression of tissue inhibitors of metalloproteinase (TIMP-2) compared to the other treatment and control groups. To conclude, we demonstrate that pirenzepine may improve the prognosis of monochromatic light-induced myopia in guinea pigs, possibly by both regulating the balance of MMP-2 and TIMP-2 in sclera and increasing the TH expression in retina.
Related JoVE Video
Oxidation of ascorbic acid by a (salen)ruthenium(vi) nitrido complex in aqueous solution.
Chem. Commun. (Camb.)
PUBLISHED: 11-06-2014
Show Abstract
Hide Abstract
The oxidation of ascorbic acid (H2A) by [Ru(VI)(N)(L)(MeOH)](+) in aqueous acidic solutions has the following stoichiometry: 2[Ru(VI)(N)] + 3H2A ? 2[Ru(III)(NH2-HA)](+) + A. Mechanisms involving HAT/N-rebound at low pH (?2) and nucleophilic attack at the nitride at high pH (?5) are proposed.
Related JoVE Video
tert-Butyl Peroxybenzoate-Promoted ?-Methylation of 1,3-Dicarbonyl Compounds.
J. Org. Chem.
PUBLISHED: 10-31-2014
Show Abstract
Hide Abstract
A tert-butyl peroxybenzoate (TBPB)-promoted direct ?-methylation of 1,3-dicarbonyl compounds has been developed, providing ?-methyl derivatives in moderate to good yields. In this procedure, TBPB plays a dual role, serving as both the methyl source and radical initiator. This work represents a key complement to the traditional ?-methylation of 1,3-dicarbonyl compounds using methyl iodide.
Related JoVE Video
[Downregulation of 53BP1 by short hairpin RNA enhances radiosensitivity in laryngeal carcinoma cells].
Sichuan Da Xue Xue Bao Yi Xue Ban
PUBLISHED: 10-25-2014
Show Abstract
Hide Abstract
To determine the effect of downregulation of 53BP1 expression on cell growth and radiosensitivity in laryngeal carcinoma Hep-2 cells.
Related JoVE Video
[Long term follow-up and prognostic analysis of 85 cases with primary gastrointestinal diffuse large B cell lymphoma].
Zhonghua Xue Ye Xue Za Zhi
PUBLISHED: 10-24-2014
Show Abstract
Hide Abstract
To analyze the clinical characteristics, prognostic factors in patients with primary gastrointestinal diffuse large B cell lymphoma (PGI-DLBCL).
Related JoVE Video
Testing of MERS-CoV replication inhibitors for their ability to block viral entry.
Antimicrob. Agents Chemother.
PUBLISHED: 10-22-2014
Show Abstract
Hide Abstract
As of 23 July 2014, 837 laboratory-confirmed cases 19 es of MERS-CoV infection, including 291 deaths, had been reported to WHO (http://www.who.int/csr/disease/coronavirus_infections/en/), raising concerns about its pandemic potential and calling for the development of vaccines and therapeutics against MERS-CoV infection.…
Related JoVE Video
Establishment of a New Zealand Rabbit Model of Spinal Tuberculosis.
J Spinal Disord Tech
PUBLISHED: 10-18-2014
Show Abstract
Hide Abstract
experimental study.
Related JoVE Video
Oxalyl Amide Assisted Palladium-Catalyzed Arylation of C(sp(2))-H Bond at the ? Position.
Org. Lett.
PUBLISHED: 10-16-2014
Show Abstract
Hide Abstract
A successful protocol has been developed for ?-arylation of ?-arylethamines at the ortho position under mild conditions. The newly developed methodology first presents broad substrate scope, great functional group tolerance, and good to excellent yield in the synthesis of substituted ?-arylethylamines. The transformation represents a practical advantage of oxalyl amide in assistance with C-H functionalization at a remote position.
Related JoVE Video
[Effects of interrupted abdominal aorta compression on cardiopulmonary cerebral resuscitation after cardiac arrest in rabbit].
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue
PUBLISHED: 10-16-2014
Show Abstract
Hide Abstract
To explore the effect of the interrupted abdominal aorta compression after cardiopulmonary resuscitation (IAAC-CPR) on cardiopulmonary cerebral resuscitation in a rabbit model of cardiac arrest (CA).
Related JoVE Video
Unified strategy to monoterpene indole alkaloids: total syntheses of (±)-goniomitine, (±)-1,2-dehydroaspidospermidine, (±)-aspidospermidine, (±)-vincadifformine, and (±)-kopsihainanine a.
J. Am. Chem. Soc.
PUBLISHED: 10-13-2014
Show Abstract
Hide Abstract
Total syntheses of (±)-goniomitine, (±)-1,2-dehydroaspidospermidine, (±)-aspidospermidine, (±)-vincadifformine, and (±)-kopsihainanine A were achieved featuring two common key steps: (1) a palladium-catalyzed decarboxylative vinylation that provides quick access to cyclopentene intermediates containing all of the carbons present in the natural products and (2) an integrated oxidation/reduction/cyclization (iORC) sequence for skeletal reorganization that converts the cyclopentenes to the pentacyclic structures of the natural products. By incorporation of a geometric constraint to iORC substrates, both the chemoselectivity (C7 vs N1 cyclization) and the stereoselectivity (trans- vs cis-fused ring system) of the cyclization process can be controlled.
Related JoVE Video
Drug delivery: nanoengineered particles for enhanced intra-articular retention and delivery of proteins (adv. Healthcare mater. 10/2014).
Adv Healthc Mater
PUBLISHED: 10-11-2014
Show Abstract
Hide Abstract
Localized intra-articular delivery of anti-inflammatory proteins can reduce inflammation in osteoarthritis but poses a challenge because of rapid clearance within few hours of injection. On page 1562, A. J. García and co-workers report a new class of polymer that forms self-assembled nanoparticles with therapeutic proteins for prolonged retention in intra-articular joint spaces compared to bolus protein doses.
Related JoVE Video
Does Water-jet Force Make a Difference in Fat Grafting: in Vitro and in Vivo Evidence of Improved Lipoaspirates Viability and Fat Grafts Survival.
Plast. Reconstr. Surg.
PUBLISHED: 10-07-2014
Show Abstract
Hide Abstract
Recent literatures reveal that water-jet assisted liposuction offers a new method to conventional liposuction techniques by using the gentle spray of fluid. However, there has not yet been a systematic, randomized, controlled study to demonstrate its effect on fresh lipoaspirates' vitality and postoperative fat survival. In this study, we compared the liposuction procedure with or without water-jet assistance respectively in a blinded way.
Related JoVE Video
Charge-transfer interactions for the fabrication of multifunctional viral nanoparticles.
Chem. Commun. (Camb.)
PUBLISHED: 10-03-2014
Show Abstract
Hide Abstract
A facile strategy to fabricate multifunctional viral nanoparticles was described by introducing charge-transfer interactions between a pyrenyl motif with dinitrophenyl and pyridinium-contained guest molecules.
Related JoVE Video
Removal of elemental mercury from flue gas by thermally activated ammonium persulfate in a bubble column reactor.
Environ. Sci. Technol.
PUBLISHED: 10-03-2014
Show Abstract
Hide Abstract
In this article, a novel technique on removal of elemental mercury (Hg(0)) from flue gas by thermally activated ammonium persulfate ((NH4)2S2O8) has been developed for the first time. Some experiments were carried out in a bubble column reactor to evaluate the effects of process parameters on Hg(0) removal. The mechanism and kinetics of Hg(0) removal are also studied. The results show that the parameters, (NH4)2S2O8 concentration, activation temperature and solution pH, have significant impacts on Hg(0) removal. The parameters, Hg(0), SO2 and NO concentration, only have small effects on Hg(0) removal. Hg(0) is removed by oxidations of (NH4)2S2O8, sulfate and hydroxyl free radicals. When (NH4)2S2O8 concentration is more than 0.1 mol/L and solution pH is lower than 9.71, Hg(0) removal by thermally activated (NH4)2S2O8 meets a pseudo-first-order fast reaction with respect to Hg(0). However, when (NH4)2S2O8 concentration is less than 0.1 mol/L or solution pH is higher than 9.71, the removal process meets a moderate speed reaction with respect to Hg(0). The above results indicate that this technique is a feasible method for emission control of Hg(0) from flue gas.
Related JoVE Video
Anticancer and antimicrobial activities and chemical composition of the birch mazegill mushroom Lenzites betulina (higher Basidiomycetes).
Int J Med Mushrooms
PUBLISHED: 10-02-2014
Show Abstract
Hide Abstract
The anticancer properties, antibiotic activity, and chemical composition of Lenzites betulina ethanol extract (EE) were evaluated. Eight compounds including 5 sterols were isolated from L. betulina, and 7 compounds were isolated from L. betulina for the first time. The EE displayed strong anticancer activity against tumor cell line MDA-MB-231, with a half maximal inhibitory concentration of 51.46 ?g/mL, and there was 83.15% inhibition at a concentration of 200 ?g/mL (MTT assay). The antimicrobial activity of the EE was evaluated against 6 microorganisms-Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Fusarium graminearum, Gibberella zeae, and Cercosporella albo-maculans-and the EE showed moderate antibiotic activity. These results suggest that L. betulina could be a good anticancer and antibiotic agent.
Related JoVE Video
RP5-833A20.1/miR-382-5p/NFIA-Dependent Signal Transduction Pathway Contributes to the Regulation of Cholesterol Homeostasis and Inflammatory Reaction.
Arterioscler. Thromb. Vasc. Biol.
PUBLISHED: 09-30-2014
Show Abstract
Hide Abstract
Cardiovascular disease caused by atherosclerosis is the number one cause of death in Western countries and threatens to become the major cause of morbidity and mortality worldwide. Long noncoding RNAs are emerging as new players in gene regulation, but how long noncoding RNAs operate in the development of atherosclerosis remains unclear.
Related JoVE Video
Analyte migration electrospray ionization for rapid analysis of complex samples with small volume using mass spectrometry.
Analyst
PUBLISHED: 09-30-2014
Show Abstract
Hide Abstract
Complex biological matrices can be effectively removed in the analyte migration process because of their weak solubility in organic solvents. This technique offers other potential advantages, including high-sensitivity and the capability of performing a direct analysis of the components and their metabolites in small-volume raw samples without any pretreatment.
Related JoVE Video
[Evaluating the missing heritability of bipolar disorder using the multifactorial liability threshold model].
Yi Chuan
PUBLISHED: 09-25-2014
Show Abstract
Hide Abstract
In order to evaluate the missing heritability of bipolar disorder, we queried the GWAS catalog of National Human Genome Research Institute, retrieve all the susceptible gene variation of bipolar disorder, and calculate the heritability explanation degree of each susceptibility variant using the multifactorial liability threshold model. The total heritability explanation degree of bipolar disorder was obtained through summing up the heritability explanation degree of each susceptibility variant. Then, we evaluated the missing heritability of bipolar disorder based on the total heritability explanation degree. The results showed that the total heritability explanation degree of bipolar disorder explained by known susceptible variants was 38.34%, and the other 61.66% of heritability can't be explained by known susceptibility variants, which belong to the missing heritability of bipolar disorder. The total heritability explanation degree of bipolar disorder in this study was significantly increased compared to earlier similar studies abroad. With constant discovery of new bipolar disorder susceptibility variants, the missing heritability of bipolar disorder has been greatly reduced, but the missing heritability of bipolar disorder still exists and occupies a large part of the bipolar disorder heritability, indicating that the molecular genetic mechanisms of bipolar disorder need to be further clarified.
Related JoVE Video
Cardioprotective effects of single oral dose of nicorandil before selective percutaneous coronary intervention.
Anadolu Kardiyol Derg
PUBLISHED: 09-25-2014
Show Abstract
Hide Abstract
Nicorandil, an opener of ATP-sensitive K+ channels, was used to treat angina in patients with coronary artery disease. In this study, we aim to investigate the cardioprotective effects of single oral dose of nicorandil in patients undergoing selective percutaneous coronary intervention (PCI).
Related JoVE Video
A Nanoparticle-Encapsulated non-Nucleoside Reverse-Transcriptase Inhibitor with Enhanced Anti-HIV-1 Activity and Prolonged Circulation Time in Plasma.
Curr. Pharm. Des.
PUBLISHED: 09-21-2014
Show Abstract
Hide Abstract
Non-nucleoside reverse-transcriptase inhibitors (NNRTIs), major components of highly active antiretroviral therapy (HAART), are effective in suppressing viral replication and preventing the progress of HIV-1 infection to AIDS. However, rapid blood clearance in vivo could significantly impair the efficiency of the anti-HIV-1 activity and result in multiple daily doses which might lead to poor patient compliance. Here we attempted to employ biodegradable organic nanoparticles (NPs) to encapsulate DAAN15h, a derivative of 4-substituted 1,5-diarylaniline with potent anti-HIV activities. Nanoparticles encapsulating DAAN15h (NP-DAAN15h) displayed a spherical shape with a size of 97.01 ± 3.64 nm and zeta potential of -19.1 ± 3.78 mV, and they exhibited a sustained controlled release behavior in vitro. The cellular uptake of NPs on TZM-b1 cells, MT-2 cells and M7 cells, possibly through lipid raft-mediated and energy-dependent active transport processes, was significantly enhanced. NP-DAAN15h, which possessed no significant in vitro cytotoxicity, showed improved antiviral activity against laboratory-adapted and primary HIV-1 isolates with different subtypes and tropisms, including RT-resistant variants. NP-DAAN15h exhibited a significantly prolonged blood circulation time, decreased plasma elimination rate, and enhanced AUC(0-t). NP-DAAN15h, a nanoparticle-encapsulated NNRTI, exhibits enhanced cellular uptake, improved anti-HIV-1 efficacy and prolonged in vivo circulation time, suggesting good potential for further development as a new NNRTI formulation for clinical use.
Related JoVE Video
New KF-PP-SVM classification method for EEG in brain-computer interfaces.
Biomed Mater Eng
PUBLISHED: 09-18-2014
Show Abstract
Hide Abstract
Classification methods are a crucial direction in the current study of brain-computer interfaces (BCIs). To improve the classification accuracy for electroencephalogram (EEG) signals, a novel KF-PP-SVM (kernel fisher, posterior probability, and support vector machine) classification method is developed. Its detailed process entails the use of common spatial patterns to obtain features, based on which the within-class scatter is calculated. Then the scatter is added into the kernel function of a radial basis function to construct a new kernel function. This new kernel is integrated into the SVM to obtain a new classification model. Finally, the output of SVM is calculated based on posterior probability and the final recognition result is obtained. To evaluate the effectiveness of the proposed KF-PP-SVM method, EEG data collected from laboratory are processed with four different classification schemes (KF-PP-SVM, KF-SVM, PP-SVM, and SVM). The results showed that the overall average improvements arising from the use of the KF-PP-SVM scheme as opposed to KF-SVM, PP-SVM and SVM schemes are 2.49%, 5.83 % and 6.49 % respectively.
Related JoVE Video
[Potential medical applications of nanoscale particles of viruses].
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi
PUBLISHED: 09-16-2014
Show Abstract
Hide Abstract
The study of viruses traditionally focused on their roles as infectious agents and as tools for understanding cell biology. Recently, however, with the development of structural biology, viruses have now been receiving particular attention in nanotechnology. By chemical methods or by gene modification, viruses have been functionalized as potential building blocks for several applications, such as drug/gene delivery vehicles, advanced vaccine vehicles, and special inorganic or organic nanomaterials. Here we highlight some of the recent progresses in the medical applications of viruses.
Related JoVE Video
Expression of polycystins and fibrocystin on primary cilia of lung cells.
Biochem. Cell Biol.
PUBLISHED: 09-12-2014
Show Abstract
Hide Abstract
Mutations in polycystin-1, polycystin-2, or fibrocystin account for autosomal dominant or recessive polycystic kidney disease. Renal cystogenesis is linked to abnormal localization and function of these cystoproteins in renal primary cilia. They are also expressed in extrarenal tissues in which their functions are unclear. Here we found that human type-II alveolar epithelial A549, airway submucosal Calu-3 cells, and rat bronchioles contain primary or multiple cilia in which we detected these cystoproteins. At sub-confluency, polycystin-1 was expressed on plasma membrane, while polycystin-2 was localized to the ER of resting cells. Both polycystins were detected on the spindle and mid-body of mitotic cells, while fibrocystin was on centrosome throughout cell cycle. Polycystins and fibrocystin may participate in regulating mucociliary sensing and transport within pulmonary airways.
Related JoVE Video
A robust electrode configuration for bioimpedance measurement of respiration.
J Healthc Eng
PUBLISHED: 09-07-2014
Show Abstract
Hide Abstract
Electrode configuration is an important issue in the continuous measurement of respiration using impedance pneumography (IP). The robust configuration is usually confirmed by comparing the amplitude of the IP signals acquired with different electrode configurations, while the relative change in waveform and the effects of body posture and respiratory pattern are ignored. In this study, the IP signals and respiratory volume are simultaneously acquired from 8 healthy subjects in supine, left lying, right lying and prone postures, and the subjects are asked to perform four respiratory patterns including free breathing, thoracic breathing, abdominal breathing and apnea. The IP signals are acquired with four different chest electrode configurations, and the volume are measured using pneumotachograph (PNT). Differences in correlation and absolute deviation between the IP-derived and PNT-derived respiratory volume are assessed. The influences of noise, respiratory pattern and body posture on the IP signals of different configurations have significant difference (p < 0.05). The robust electrode configuration is found on the axillary midline, which is suitable for long term respiration monitoring.
Related JoVE Video
Copper-mediated/catalyzed oxyalkylation of alkenes with alkylnitriles.
Chemistry
PUBLISHED: 09-02-2014
Show Abstract
Hide Abstract
A copper-promoted oxyalkylation of alkenes with alkylnitriles has been developed. The protocol provides rapid access to phthalides (?-lactones) or isochromanones (?-lactones) via the formation of a C(sp(3) )?C(sp(3) ) and a C(sp(3) )?O bond with the generation of up to two quaternary carbon atoms. Mechanistic studies suggest that this reaction is initiated by the formation of the C(sp(3) )?C(sp(3) ) bond rather than the C(sp(3) )?O bond. Catalytic conditions were subsequently developed using carboxylic acid as an internal nucleophile.
Related JoVE Video
[P38 MAPK signaling pathway regulates nuclear factor-?B and inducible nitric oxide synthase expressions in the substantia nigra in a mouse model of Parkinson's disease].
Nan Fang Yi Ke Da Xue Xue Bao
PUBLISHED: 09-02-2014
Show Abstract
Hide Abstract
To investigate the role of P38 mitogen-activated protein kinase (P38 MAPK) signaling pathway in regulating the expression of nuclear factor-?B (NF-?B) and inducible nitric oxide synthase (iNOS) in the substantia nigra (SN) of a mouse model of Parkinson's disease (PD).
Related JoVE Video
Oroxylin A inhibits ATRA-induced IL-6 expression involved in retinoic acid syndrome by down-regulating CHOP.
Gene
PUBLISHED: 09-01-2014
Show Abstract
Hide Abstract
Production of IL-6 constituted the major cause of death in the ATRA trial called retinoic acid syndrome (RAS). LAP and LIP are active and inactive isoforms of C/EBP?, respectively. Inactive LIP dimerized with LAP to eliminate its activity. Following treatment with ATRA, CHOP expression was increased and dimerized with LIP more preferentially than LAP to rescue function of LAP. Oroxylin A has been reported to activate CHOP, a key mediator of unfolded protein response (UPR) pathway, and resulted in apoptosis. Interestingly, we found that low concentration of oroxylin A (? 40 ?M) showed no apoptosis effect on NB4 and HL-60 cells and decreased the CHOP protein level via promoting its degradation. MG132 was utilized to conform the effect of oroxylin A on degrading CHOP. Our results showed that oroxylin A decreased the level of IL-6 secretion of NB4 cells with or without ATRA treatment while the effect was eliminated by C/EBP? siRNA. We conclude that oroxylin A possessed abilities of inhibiting the ATRA-induced IL-6 production via modulation of LAP/LIP/CHOP in leukemia cell lines, which could providing a therapeutic strategy for RAS.
Related JoVE Video
Association between raf kinase inhibitor protein loss and prognosis in cancers of the digestive system: a meta-analysis.
Cancer Biomark
PUBLISHED: 08-30-2014
Show Abstract
Hide Abstract
Loss of Raf kinase inhibitor protein (RKIP) may contribute to metastasis in a variety of human cancers. Many studies have evaluated whether loss of RKIP expression is a prognostic factor for survival in cancers of the digestive system, however, its predictive value remains controversial. Thus, we performed a meta-analysis to obtain a more comprehensive estimate of the prognostic value of RKIP expression in digestive system cancers.
Related JoVE Video
[Progress on bone marrow mesenchymal stem cells transplantation for spinal cord injury].
Zhongguo Gu Shang
PUBLISHED: 08-30-2014
Show Abstract
Hide Abstract
Bone marrow mesenchymal cells (BMSCs) are regarded as donor cells in cell transplantation therapies for spinal cord injury (SCI) for they have the ability of favourable proliferation and multi-directional differentiation, and are easily isolated and culturd and have less immunological reaction. It has been confirmed that subarachnoid space injection is the most ideal delivery technique of BMSCs. Bone marrow mesenchymal stem cells transplantation is safe and its reconditioning role is certain for SCI in early clinical application. The mechanism of BMSCs promoting functional recovery after SCI is probably concerned with vicarious function, nerve trophism, immunosuppression and promoting axonal regeneration by BMSCs.
Related JoVE Video
Tumor suppressor microRNA-27a in colorectal carcinogenesis and progression by targeting SGPP1 and Smad2.
PLoS ONE
PUBLISHED: 08-28-2014
Show Abstract
Hide Abstract
The aberrant expression of microRNAs (miRNAs) is associated with colorectal carcinogenesis, but the underlying mechanisms are not clear. This study showed that the miRNA-27a (miR-27a) was significantly reduced in colorectal cancer tissues and colorectal cancer cell lines, and that the reduced miR-27a was associated with distant metastasis and colorectal cancer clinical pathological stages-miR-27a was lower at stages III/IV than that at stage II. Bioinformatic and systemic biological analysis predicted several targets of miR-27a, among them SGPP1 and Smad2 were significantly affected. SGPP1 and Smad2 at mRNA and protein levels were negatively correlated with miR-27a in human colorectal cancer tissues and cancer cell lines. Increased miR-27a significantly repressed SGPP1 and Smad2 at transcriptional and translational levels. Functional studies showed that increasing miR-27a inhibited colon cancer cell proliferation, promoted apoptosis and attenuated cell migration, which were also linked to downregulation of p-STAT3 and upregulation of cleaved caspase 3. In vivo, miR-27a inhibited colon cancer cell growth in tumor-bearing mice. Taken together, this study has revealed miR-27a as a tumor suppressor and has identified SGPP1 and Smad2 as novel targets of miR-27a, linking to STAT3 for regulating cancer cell proliferation, apoptosis and migration in colorectal cancer. Therefore, miR-27a could be a useful biomarker for monitoring colorectal cancer development and progression, and also could have a therapeutic potential by targeting SGPP1, Smad2 and STAT3 for colorectal cancer therapy.
Related JoVE Video
Conjugation of a nonspecific antiviral sapogenin with a specific HIV fusion inhibitor: a promising strategy for discovering new antiviral therapeutics.
J. Med. Chem.
PUBLISHED: 08-26-2014
Show Abstract
Hide Abstract
Triterpene saponins are a major group of active components in natural products with nonspecific antiviral activities, while T20 peptide (enfuvirtide), which contains a helix zone-binding domain (HBD), is a gp41-specific HIV-1 fusion inhibitor. In this paper, we report the design, synthesis, and structure-activity relationship (SAR) of a group of hybrid molecules in which bioactive triterpene sapogenins were covalently attached to the HBD-containing peptides via click chemistry. We found that either the triterpenes or peptide part alone showed weak activity against HIV-1 Env-mediated cell-cell fusion, while the hybrids generated a strong cooperative effect. Among them, P26-BApc exhibited anti-HIV-1 activity against both T20-sensitive and -resistant HIV-1 strains and improved pharmacokinetic properties. These results suggest that this scaffold design is a promising strategy for developing new HIV-1 fusion inhibitors and possibly novel antiviral therapeutics against other viruses with class I fusion proteins.
Related JoVE Video
Persistent hepatitis C virus infections and hepatopathological manifestations in immune-competent humanized mice.
Cell Res.
PUBLISHED: 08-26-2014
Show Abstract
Hide Abstract
The majority of hepatitis C virus (HCV) infection develops chronic infection, which causes steatosis, cirrhosis and hepatocellular carcinoma. However, understanding HCV chronicity and pathogenesis is hampered by its narrow host range, mostly restricted to human and chimpanzee. Recent endeavour to infect a variety of humanized mice has not been able to achieve persistent HCV infection unless the essential innate immune responsive genes are knocked out. Nevertheless, such immune-compromised humanized mice still lacked HCV infection-induced hepatopathogenesis. Here we report that transgenic mice in ICR background harboring both human CD81 and occludin genes (C/O(Tg)) are permissive to HCV infection at a chronicity rate comparable to humans. In this mouse model, HCV accomplishes its replication cycle, leading to sustained viremia and infectivity for more than 12 months post infection with expected fibrotic and cirrhotic progression. Host factors favorable for HCV replication, and inadequate innate immune-response may contribute to the persistence. Lastly, NS3/4 protease inhibitor telaprevir can effectively inhibit de novo RNA synthesis and acute HCV infection of C/O(Tg) mice. Thus, chronic HCV infection with complete replication cycle and hepatopathologic manifestations is recapitulated, for the first time, in immune-competent mice. This model will open a new venue to study the mechanisms of chronic hepatitis C and develop better treatments.
Related JoVE Video
Wogonin induces cell cycle arrest and erythroid differentiation in imatinib-resistant K562 cells and primary CML cells.
Oncotarget
PUBLISHED: 08-24-2014
Show Abstract
Hide Abstract
Wogonin, a flavonoid derived from Scutellaria baicalensis Georgi, has been demonstrated to be highly effective in treating hematologic malignancies. In this study, we investigated the anticancer effects of wogonin on K562 cells, K562 imatinib-resistant cells, and primary patient-derived CML cells. Wogonin up-regulated transcription factor GATA-1 and enhanced binding between GATA-1 and FOG-1, thereby increasing expression of erythroid-differentiation genes. Wogonin also up-regulated the expression of p21 and induced cell cycle arrest. Studies employing benzidine staining and analyses of cell surface markers glycophorin A (GPA) and CD71 indicated that wogonin promoted differentiation of K562, imatinib-resistant K562, and primary patient-derived CML cells. Wogonin also enhanced binding between GATA-1 and MEK, resulting in inhibition of the growth of CML cells. Additionally, in vivo studies showed that wogonin decreased the number of CML cells and prolonged survival of NOD/SCID mice injected with K562 and imatinib-resistant K562 cells. These data suggested that wogonin induces cycle arrest and erythroid differentiation in vitro and inhibits proliferation in vivo.
Related JoVE Video
[The prevalence and clinical significance of arthritis in patients with systemic sclerosis].
Zhonghua Nei Ke Za Zhi
PUBLISHED: 08-23-2014
Show Abstract
Hide Abstract
To explore the prevalence and independent influencing factors of arthritis in Chinese patients with systemic sclerosis (SSc).
Related JoVE Video
[The clinical characteristics of polyserositis as main presentation of chronic graft-versus-host rejection disease after allogeneic hematopoietic stem cell transplantation].
Zhonghua Nei Ke Za Zhi
PUBLISHED: 08-23-2014
Show Abstract
Hide Abstract
To analyze the clinical characteristics of polyserositis associated with chronic graft-versus-host disease (cGVHD) after allogeneic hematopoietic transplantation(allo-HSCT).
Related JoVE Video
[The clinical characteristics of systemic sclerosis-related pulmonary arterial hypertension].
Zhonghua Nei Ke Za Zhi
PUBLISHED: 08-23-2014
Show Abstract
Hide Abstract
To study the clinical, cardiopulmonary functional and hemodynamic profiles of systemic sclerosis patients with pulmonary hypertension (SSc-PAH) compared with those of idiopathic pulmonary hypertension (IPAH).
Related JoVE Video
Simvastatin to modify neutrophil function in older patients with septic pneumonia (SNOOPI): study protocol for a randomised placebo-controlled trial.
Trials
PUBLISHED: 08-22-2014
Show Abstract
Hide Abstract
Community-acquired pneumonia (CAP) is considered the leading cause of death from infectious disease in developed countries, while complications of CAP - sepsis being the most common and challenging - increase the risk of mortality. During the progression of sepsis, a state of neutrophil 'paralysis' develops resulting in the impairment of neutrophil anti-microbial functions including: chemotaxis, production of reactive oxygen species, and formation of neutrophil extracellular traps (NETs). Mechanisms underlying defective neutrophil function remain elusive although NET formation has been implicated in the immunosuppression and increased rates of sepsis observed in neonates. There is, however, increasing evidence that statins are able to modulate neutrophil function in sepsis as several systematic reviews have concluded that statins have a role in improving infection-related outcomes and mortality while, in vitro, statins have also been shown to boost NET formation in healthy individuals.
Related JoVE Video
Tuning electronic and magnetic properties of silicene with magnetic superhalogens.
Phys Chem Chem Phys
PUBLISHED: 08-22-2014
Show Abstract
Hide Abstract
Due to its compatibility with the well-developed Si-based semiconductor industry, silicene has attracted considerable attention. Using density functional theory we show for the first time that the recently synthesized superhalogen MnCl3 can be used to tune the electronic and magnetic properties of silicene, from semi-metallic to semiconducting with a wide range of band gaps, as well as from nonmagnetic to ferromagnetic (or antiferromagnetic) by changing the coverage of the superhalogen molecules. The electronic properties can be further modulated when a superhalogen and a halogen are used synergistically. The present study indicates that because of the large electron affinity and rich structural diversity superhalogen molecules have advantages over the conventional halogen atoms in modulating the material properties of silicene.
Related JoVE Video
Vacuolin-1 potently and reversibly inhibits autophagosome-lysosome fusion by activating RAB5A.
Autophagy
PUBLISHED: 08-20-2014
Show Abstract
Hide Abstract
Autophagy is a catabolic lysosomal degradation process essential for cellular homeostasis and cell survival. Dysfunctional autophagy has been associated with a wide range of human diseases, e.g., cancer and neurodegenerative diseases. A large number of small molecules that modulate autophagy have been widely used to dissect this process and some of them, e.g., chloroquine (CQ), might be ultimately applied to treat a variety of autophagy-associated human diseases. Here we found that vacuolin-1 potently and reversibly inhibited the fusion between autophagosomes and lysosomes in mammalian cells, thereby inducing the accumulation of autophagosomes. Interestingly, vacuolin-1 was less toxic but at least 10-fold more potent in inhibiting autophagy compared with CQ. Vacuolin-1 treatment also blocked the fusion between endosomes and lysosomes, resulting in a defect in general endosomal-lysosomal degradation. Treatment of cells with vacuolin-1 alkalinized lysosomal pH and decreased lysosomal Ca (2+) content. Besides marginally inhibiting vacuolar ATPase activity, vacuolin-1 treatment markedly activated RAB5A GTPase activity. Expression of a dominant negative mutant of RAB5A or RAB5A knockdown significantly inhibited vacuolin-1-induced autophagosome-lysosome fusion blockage, whereas expression of a constitutive active form of RAB5A suppressed autophagosome-lysosome fusion. These data suggest that vacuolin-1 activates RAB5A to block autophagosome-lysosome fusion. Vacuolin-1 and its analogs present a novel class of drug that can potently and reversibly modulate autophagy.
Related JoVE Video
Predictive value of decoy receptor 3 in postoperative nosocomial bacterial meningitis.
Int J Mol Sci
PUBLISHED: 08-13-2014
Show Abstract
Hide Abstract
Nosocomial bacterial meningitis requires timely treatment, but what is difficult is the prompt and accurate diagnosis of this disease. The aim of this study was to assess the potential role of decoy receptor 3 (DcR3) levels in the differentiation of bacterial meningitis from non-bacterial meningitis. A total of 123 patients were recruited in this study, among them 80 patients being with bacterial meningitis and 43 patients with non-bacterial meningitis. Bacterial meningitis was confirmed by bacterial culture of cerebrospinal fluid (CSF) culture and enzyme-linked immunosorbent assay (ELISA) was used to detect the level of DcR3 in CSF. CSF levels of DcR3 were statistically significant between patients with bacterial meningitis and those with non-bacterial meningitis (p < 0.001). A total of 48.75% of patients with bacterial meningitis received antibiotic >24 h before CSF sampling, which was much higher than that of non-bacterial meningitis. CSF leucocyte count yielded the highest diagnostic value, with an area under the receiver operating characteristic curve (ROC) of 0.928, followed by DcR3. At a critical value of 0.201 ng/mL for DcR3, the sensitivity and specificity were 78.75% and 81.40% respectively. DcR3 in CSF may be a valuable predictor for differentiating patients with bacterial meningitis from those with non-bacterial meningitis. Further studies are needed for the validation of this study.
Related JoVE Video
Emodin suppresses pulmonary metastasis of breast cancer accompanied with decreased macrophage recruitment and M2 polarization in the lungs.
Breast Cancer Res. Treat.
PUBLISHED: 08-08-2014
Show Abstract
Hide Abstract
Breast cancer is the leading cause of death in female cancer patients due to the lack of effective treatment for metastasis. Macrophages are the most abundant immune cells in the primary and metastatic tumors, and contribute to tumor initiation, progression, and metastasis. Emodin has been found to exert anti-tumor effects through promoting cell cycle arrest and apoptosis, and inhibiting angiogenesis, but its effects on tumor-associated macrophages during cancer metastasis have not been investigated. Mice inoculated with 4T1 or EO771 breast cancer cells orthotopically were treated with Emodin after the primary tumors reached 200 mm(3) in size. Primary tumor growth, lung metastasis, and macrophage infiltration in the lungs were analyzed. In vitro experiments were performed to examine the effects of Emodin on macrophage migration and M2 polarization, and the underlying mechanisms. Emodin significantly suppressed breast cancer lung metastasis in both orthotopic mouse models without apparent effects on primary tumors. Reduced infiltration of F4/80+ macrophages and Ym1+ M2 macrophages in lungs was observed in Emodin-treated mice. In vitro experiments demonstrated that Emodin decreased the migration of macrophages toward tumor cell-conditioned medium (TCM) and inhibited macrophage M2 polarization induced by TCM. Mechanistically, Emodin suppressed STAT6 phosphorylation and C/EBP? expression, two crucial signaling events in macrophage M2 polarization, in macrophages treated with IL-4 or TCM. Taken together, our study, for the first time, demonstrated that Emodin suppressed pulmonary metastasis of breast cancer probably through inhibiting macrophage recruitment and M2 polarization in the lungs by reducing STAT6 phosphorylation and C/EBP? expression.
Related JoVE Video
Sandwichlike magnesium silicate/reduced graphene oxide nanocomposite for enhanced Pb²? and methylene blue adsorption.
ACS Appl Mater Interfaces
PUBLISHED: 08-07-2014
Show Abstract
Hide Abstract
A sandwichlike magnesium silicate/reduced graphene oxide nanocomposite (MgSi/RGO) with high adsorption efficiency of organic dye and lead ion was synthesized by a hydrothermal approach. MgSi nanopetals were formed in situ on both sides of RGO sheets. The nanocomposite with good dispersion of nanopetals exhibits a high specific surface area of 450 m(2)/g and a good mass transportation property. Compared to MgSi and RGO, the mechanical stability and adsorption capacity of the nanocomposite is significantly improved due to the synergistic effect. The maximum adsorption capacities for methylene blue and lead ion are 433 and 416 mg/g, respectively.
Related JoVE Video
A mini-electrochemical system integrated micropipet tip and pencil graphite electrode for detection of anticancer drug sensitivity in vitro.
Biosens Bioelectron
PUBLISHED: 07-20-2014
Show Abstract
Hide Abstract
Developing a reliable and cost-effective miniaturized electroanalysis tool is of vital importance for cell electrochemical analysis. In this work, a novel mini-electrochemical system has been constructed for trace detection of cell samples. The mini-electrochemical system was constructed by integrating a pencil graphite modified by threonine (PT/PGE) as working electrode, an Ag/AgCl (Sat'd) as reference electrode, platinum wire as counter electrode and a micropipet tip as electrochemical cell. The mini-electrochemical system not only saved dramatically usage of samples from 500?L in traditional electrochemical system to 10?L, but also possessed an adjustable active surface area by changing the length of PT/PGE immersed into the cell suspension from 3mm to 15mm, and the linear equation was ipa=2.25l-2.64 (R(2)=0.990). The system was successfully used in detection of MCF-7 cells, and a nonlinear exponent relationship between peak current and the cell number range from 3.0×l0(3) to 7.0×l0(6)cellsmL(-1) was established firstly with the index equation ipa=59.557e (-C/1.709)-71.486 (R(2)=0.954). Finally, the system was used for evaluating the sensitivity of cyclophosphamide on MCF-7 cell, and the result was corresponded well with that of MTT assay. The proposed system is sufficiently simple, cheap and easy operated, and could be applied in electrochemical detection of other biological samples.
Related JoVE Video
Next generation sequencing unravels the biosynthetic ability of Spearmint ( Mentha spicata ) peltate glandular trichomes through comparative transcriptomics.
BMC Plant Biol.
PUBLISHED: 07-09-2014
Show Abstract
Hide Abstract
BackgroundPlant glandular trichomes are chemical factories with specialized metabolic capabilities to produce diverse compounds. Aromatic mint plants produce valuable essential oil in specialised glandular trichomes known as peltate glandular trichomes (PGT). Here, we performed next generation transcriptome sequencing of different tissues of Mentha spicata (spearmint) to identify differentially expressed transcripts specific to PGT. Our results provide a comprehensive overview of PGT¿s dynamic metabolic activities which will help towards pathway engineering.ResultsSpearmint RNAs from 3 different tissues: PGT, leaf and leaf stripped of PGTs (leaf-PGT) were sequenced by Illumina paired end sequencing. The sequences were assembled de novo into 40,587 non-redundant unigenes; spanning a total of 101 Mb. Functions could be assigned to 27,025 (67%) unigenes and among these 3,919 unigenes were differentially expressed in PGT relative to leaf - PGT. Lack of photosynthetic transcripts in PGT transcriptome indicated the high levels of purity of isolated PGT, as mint PGT are non-photosynthetic. A significant number of these unigenes remained unannotated or encoded hypothetical proteins. We found 16 terpene synthases (TPS), 18 cytochrome P450s, 5 lipid transfer proteins and several transcription factors that were preferentially expressed in PGT. Among the 16 TPSs, two were characterized biochemically and found to be sesquiterpene synthases.ConclusionsThe extensive transcriptome data set renders a complete description of genes differentially expressed in spearmint PGT. This will facilitate the metabolic engineering of mint terpene pathway to increase yield and also enable the development of strategies for sustainable production of novel or altered valuable compounds in mint.
Related JoVE Video
Formula Optimization of the Jiashitang Scar Removal Ointment and Antiinflammatory Compounds Screening by NF-?B Bioactivity-guided Dual-luciferase Reporter Assay System.
Phytother Res
PUBLISHED: 07-09-2014
Show Abstract
Hide Abstract
Inflammation plays a role in scar formation; therefore, decreasing inflammation benefits scar removal. Jiashitang scar removal ointment (JST) is a commercially available traditional Chinese medicinal formulation. It is composed of extracts from Carthamus tinctorius L. (Car), Rheum officinale Baill. (Rhe), Salvia miltiorrhiza Beg. (Sal), and Panax notoginseng (Burk.) F.?H. Chen (Pan), which are all herbs with potent antiinflammatory activities. Our aims are to optimize the formula of JST and to elucidate its antiinflammatory active components. Response surface methodology was applied to optimize proportions of the four herb extracts. The antiinflammatory effects were evaluated using in vitro and in vivo models. To screen for active components in this formula, a bioactivity-based ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry analysis was performed. After optimization, the antiinflammatory effects of the new formula were significantly superior to the original one. Screening identified 13 active ingredients: a series of saffiomin, emodin, salvianolic acid, tanshinone, and triterpenoid saponin derivatives. These active ingredients were predicted to exert nuclear factor-?B inhibiting effects through MAPK, PI3K/AKT, and NIK-IKK pathways. In conclusion, the original formula was successfully optimized with more potent antiinflammatory activity. These methods can be applied to researches of other formulas. Copyright © 2014 John Wiley & Sons, Ltd.
Related JoVE Video
Glucocorticoid Receptor ? Acts as a Co-activator of T-Cell Factor 4 and Enhances Glioma Cell Proliferation.
Mol. Neurobiol.
PUBLISHED: 07-07-2014
Show Abstract
Hide Abstract
We previously reported that glucocorticoid receptor ? (GR?) regulates injury-mediated astrocyte activation and contributes to glioma pathogenesis via modulation of ?-catenin/T-cell factor/lymphoid enhancer factor (TCF/LEF) transcriptional activity. The aim of this study was to characterize the mechanism behind cross-talk between GR? and ?-catenin/TCF in the progression of glioma. Here, we reported that GR? knockdown reduced U118 and Shg44 glioma cell proliferation in vitro and in vivo. Mechanistically, we found that GR? knockdown decreased TCF/LEF transcriptional activity without affecting ?-catenin/TCF complex. Both GR? and GR? directly interact with TCF-4, while only GR? is required for sustaining TCF/LEF activity under hormone-free condition. GR? bound to the N-terminus domain of TCF-4 its influence on Wnt signaling required both ligand- and DNA-binding domains (LBD and DBD, respectively). GR? and TCF-4 interaction is enough to maintain the TCF/LEF activity at a high level in the absence of ?-catenin stabilization. Taken together, these results suggest a novel cross-talk between GR? and TCF-4 which regulates Wnt signaling and the proliferation in gliomas.
Related JoVE Video
Neutrophils sense microbe size and selectively release neutrophil extracellular traps in response to large pathogens.
Nat. Immunol.
PUBLISHED: 07-01-2014
Show Abstract
Hide Abstract
Neutrophils are critical for antifungal defense, but the mechanisms that clear hyphae and other pathogens that are too large to be phagocytosed remain unknown. We found that neutrophils sensed microbe size and selectively released neutrophil extracellular traps (NETs) in response to large pathogens, such as Candida albicans hyphae and extracellular aggregates of Mycobacterium bovis, but not in response to small yeast or single bacteria. NETs were fundamental in countering large pathogens in vivo. Phagocytosis via dectin-1 acted as a sensor of microbe size and prevented NET release by downregulating the translocation of neutrophil elastase (NE) to the nucleus. Dectin-1 deficiency led to aberrant NET release and NET-mediated tissue damage during infection. Size-tailored neutrophil responses cleared large microbes and minimized pathology when microbes were small enough to be phagocytosed.
Related JoVE Video
Population transcriptomics reveals a potentially positive role of expression diversity in adaptation.
J Integr Plant Biol
PUBLISHED: 06-29-2014
Show Abstract
Hide Abstract
While it is widely accepted that genetic diversity determines the potential of adaptation, the role that gene expression variation plays in adaptation remains poorly known. Here we show that gene expression diversity could have played a positive role in the adaptation of Miscanthus lutarioriparius. RNA-seq was conducted for 80 individuals of the species, with half planted in the energy crop domestication site and the other half planted in the control site near native habitats. A leaf reference transcriptome consisting of 18 503 high-quality transcripts was obtained using a pipeline developed for de novo assembling with population RNA-seq data. The population structure and genetic diversity of M. lutarioriparius were estimated based on 30 609 genic SNPs. Population expression (Ep ) and expression diversity (Ed ) were defined to measure the average level and the magnitude of variation of a gene expression in the population, respectively. It was found that expression diversity increased while genetic diversity decreased after the species was transplanted from the native habitats to the harsh domestication site, especially for genes involved in abiotic stress resistance, histone methylation, and biomass synthesis under water limitation. The increased expression diversity could have enriched phenotypic variation directly subject to selections in the new environment.
Related JoVE Video
Recombination of Thermo-Alkalistable, High Xylooligosaccharides Producing Endo-Xylanase from Thermobifida fusca and Expression in Pichia pastoris.
Appl. Biochem. Biotechnol.
PUBLISHED: 06-26-2014
Show Abstract
Hide Abstract
For xylooligosaccharide (XO) production, endo-xylanase from Thermobifida fusca was modified by error-prone PCR and DNA shuffling. The G4SM1 mutant (S62T, S144C, N198D, and A217V) showed the most improved hydrolytic activity and was two copies expressed in Pichia pastoris under the control of GAP promoter. The maximum xylanase activity in culture supernatants was 165?±?5.5 U/ml, and the secreted protein concentration reached 493 mg/l in a 2-l baffled shake flask. After 6× His-tagged protein purification, the specific activity of G4SM1 was 2036?±?45.8 U/mg, 2.12 times greater than that of wild-type enzyme. Additionally, G4SM1 was stable over a wide pH range from 5.0 to 9.0. Meanwhile, half-life of G4SM1 thermal inactivation at 70 °C increased 8.5-fold. Three-dimensional structures suggest that two amino acid substitutions, S62T and S144C, located at catalytic domain may be responsible for the enhanced activity and thermostability of xylanase. Xylobiose was the dominant end product of xylan hydrolysis by G4SM1. Due to its attractive biochemical properties, G4SM1 has potential value in commercial XO production.
Related JoVE Video
Purtscher-like Retinopathy in Systemic Lupus Erythematosus.
Am. J. Ophthalmol.
PUBLISHED: 06-23-2014
Show Abstract
Hide Abstract
To investigate clinical characteristics of Purtscher-like retinopathy and its clinical implications among patients with systemic lupus erythematosus (SLE).
Related JoVE Video
Platelet-Inspired Multiscaled Cytophilic Interfaces with High Specificity and Efficiency toward Point-of-Care Cancer Diagnosis.
Small
PUBLISHED: 05-29-2014
Show Abstract
Hide Abstract
Inspired by the interactions between platelets and tumor cells, multiscaled cytophilic interfaces are fabricated by assembling platelet-mimicking microspheres via oxidative polymerization of polyaniline nanohairs through transverse nanochannels in the shells of hollow polystyrene microspheres, and modifying specific recognition molecules. This interface realizes efficient tumor-cell recognition and isolation from artificial blood, suggesting a new platform for metastasis diagnosis and cancer treatment.
Related JoVE Video
HLA Disparity is not crucial for the survival rate and severity of chronic health conditions in adult recipients following family donor hematopoietic stem cell transplantation.
Int. J. Hematol.
PUBLISHED: 05-05-2014
Show Abstract
Hide Abstract
The shortage of HLA-identical siblings or unrelated donors has restricted the application of hematopoietic stem cell transplantation (HSCT). Few studies have systematically assessed survival and chronic health conditions (CHCs) in the same cohort of patients after HLA-mismatched/haploidentical (mismatched) family donor transplantation. In the present study, we retrospectively analyzed the survival of 127 adult patients receiving either HLA-matched (71 cases) or HLA-mismatched (56 cases) family donor transplantation. Of 127 patients, 81 patients survived at least 2 years after HSCT and were still alive until the present investigation. We evaluated the CHCs in 76 survivors (41 matched and 35 mismatched). CHC-related information was scored according to the Bone Marrow Transplant Survivor Study questionnaire. There was no significant difference in overall survival or disease-free survival between HLA-matched and -mismatched transplant recipients. The CHCs were less severe in HLA-mismatched recipients than in matched cohorts. Multivariate analysis identified that age over 40 years at transplantation and presence of chronic graft-versus-host disease were independent risk factors for CHCs, while anti-thymocyte globulin-containing conditioning regimens might be protective. However, HLA disparity was not crucial for either the survival rate or CHCs. In conclusion, HLA-mismatched family donor transplantation can achieve comparable therapeutic effects to HLA-identical sibling transplantation.
Related JoVE Video
Clinical and laboratory characteristics of systemic sclerosis patients with pulmonary arterial hypertension in China.
Clin. Exp. Rheumatol.
PUBLISHED: 04-27-2014
Show Abstract
Hide Abstract
Pulmonary arterial hypertension (PAH) is associated with significant morbidity and mortality, especially in systemic sclerosis (SSc). Since there was no published study regarding PAH in the Chinese SSc population, we aimed to describe a cohort to provide some data for early diagnosis.
Related JoVE Video
The impact of hepatitis B virus x protein and microRNAs in hepatocellular carcinoma: a comprehensive analysis.
Tumour Biol.
PUBLISHED: 04-24-2014
Show Abstract
Hide Abstract
microRNAs (miRNAs) are evolutionarily conserved small non-coding RNAs, approximately 22 nucleotides (nts) in length, widely found in animals, plants, and viruses. Mature miRNAs control gene expression at a post-transcriptional level through blocking protein translation or inducing mRNA degradation. Many recent studies have shown that hepatitis B virus x protein (HBx), a viral protein with a crucial role in hepatogenesis, is associated with the regulation of miRNAs. This interaction impacts fundamental tumor processes, such as cell proliferation, differentiation, and apoptosis. In this review, we summarized the recent literature on the roles of HBx-regulated miRNAs in the pathogenesis of hepatocellular carcinoma.
Related JoVE Video
Role of TREM-1 in response to Aspergillus fumigatus infection in corneal epithelial cells.
Int. Immunopharmacol.
PUBLISHED: 04-21-2014
Show Abstract
Hide Abstract
Triggering receptor expressed on myeloid cells-1 (TREM-1) is a cell surface receptor that is highly expressed in inflammatory lesions caused by infectious agents such as bacteria and fungi and amplifies immune responses. The aim of the current study was to investigate TREM-1 expression in corneal epithelial cells infected by Aspergillus fumigatus (A. fumigatus) and evaluate its role. In this study, infection with A. fumigatus upregulated TREM-1 expression in corneal epithelial cells both in vitro and in vivo. Furthermore, treatment with the antagonistic peptide of TREM-1 decreased the levels of inflammatory cytokines that were enhanced by the fungal infection. We speculated that cross-talk occurs between TREM-1 and Toll-like receptor-4 (TLR-4) in cornea fungal infection. Inhibitors of TLR-4 and myeloid differentiation factor 88 (MyD88) could partially inhibit the upregulation of TREM-1 induced by A. fumigatus respectively. In addition, TLR-4 blockade enhanced the inhibitory effect of the antagonistic peptide of TREM-1 on A. fumigatus-induced inflammation. These findings suggest that TREM-1 plays critical roles in fungal infection, and targeting it may represent a novel therapeutic strategy for patients with fungal keratitis.
Related JoVE Video
Development and evaluation of a competitive ELISA using a monoclonal antibody for antibody detection after goose parvovirus virus-like particles (VLPs) and vaccine immunization in goose sera.
J. Virol. Methods
PUBLISHED: 04-15-2014
Show Abstract
Hide Abstract
An assay protocol based on a monoclonal antibody-based competitive enzyme-linked immunosorbent assay (MAb-based C-ELISA) for detecting antibodies against goose parvovirus (GPV) and its virus-like particles (VLPs) is described. The assay was developed using baculovirus-expressed recombinant VP2 virus-like particles (rVP2-VLPs) as antigens and a monoclonal antibody against GPV as the competitive antibody. Of the four anti-GPV MAbs that were screened, MAb 1G3 was selected as it was blocked by the GPV positive serum. Based on the distribution of percent inhibition (PI) of the known negative sera (n=225), a cut-off value was set at 36% inhibition. Using this cut-off value, the sensitivity of the assay was 93.3% and the specificity was 95.8%, as compared with the gold standard (virus neutralization assay). The rVP2-VLPs did not react with anti-sera to other goose pathogens, indicating that it is specific for the recognization of goose parvovirus antibodies. The assay was then validated with serum samples from goslings vaccinated with several VLPs (rVP1-VLPs, rVP2-VLPs, rVP3-VLPs, and rCGV-VLPs) and other vaccines (inactivated and attenuated). The C-ELISA described in this study is a sensitive and specific diagnostic test and should have wide applications for the sero-diagnosis and immunologic surveillance of GPV.
Related JoVE Video
Glucosylceramide synthase promotes Bcl-2 expression via the ERK signaling pathway in the K562/A02 leukemia drug-resistant cell line.
Int. J. Hematol.
PUBLISHED: 04-02-2014
Show Abstract
Hide Abstract
Multidrug resistance (MDR) to chemotherapeutic agents is a major obstacle to curative treatment of cancer. In various types of cancers, overexpression of glucosylceramide synthase (GCS) has been observed to be associated with MDR, thus making GCS a target for reversal of resistance. Our previous work demonstrated that GCS and Bcl-2 are co-overexpressed in the K562/A02 leukemia multidrug-resistant cell line compared with its sensitive counterpart, K562. In the present study, we investigated the effects of GCS on apoptosis in K562/A02 and the associated molecular mechanisms. Our results indicate that the inhibition of GCS caused downregulation of Bcl-2 as well as apoptosis enhancement in response to ADM via the ERK pathway, while JNK or p38 MAPK signaling appeared to play less significant roles in the regulation of apoptosis and MDR in K562/A02 cells. Targeting GCS by siRNA also enhanced ceramide accumulation, which is involved in GCS knockdown-induced inhibition of ERK activation and Bcl-2 expression levels.
Related JoVE Video
The synthesis of tenofovir and its analogues via asymmetric transfer hydrogenation.
Org. Lett.
PUBLISHED: 03-20-2014
Show Abstract
Hide Abstract
A series of tenofovir analogues with potential antiviral and immunobiologically active compounds were synthesized through an asymmetric transfer hydrogenation reaction from achiral purine derivatives. Up to 97% ee and good to excellent yields were achieved under mild conditions through short reaction steps. The present report suggests an efficient process to acquire tenofovir and its analogues.
Related JoVE Video
Dynamic changes in the hypothalamic- pituitary-adrenal axis during growth hormone therapy in children with growth hormone deficiency: a multicenter retrospective study.
J. Pediatr. Endocrinol. Metab.
PUBLISHED: 02-24-2014
Show Abstract
Hide Abstract
Abstract Objective: The objective of this study was to investigate changes in the hypothalamic-pituitary-adrenal (HPA) axis after recombinant human growth hormone (rhGH) therapy. Subjects: Subjects included children with growth hormone deficiency (GHD). Methods: We conducted a multicenter, retrospective study that assessed 72 GHD patients treated with rhGH during 6 months. Patients were classified into two groups: isolated GHD (IGHD; n=20) and multiple pituitary hormone deficiencies (MPHD; n=52). The HPA axis and other hormones were evaluated at baseline and every 3 months. Results: In the MPHD group, 32 patients had adrenocorticotrophic hormone deficiency and received hydrocortisone before rhGH therapy. In the other 20/52 MPHD patients, the cortisol (COR) level was significantly reduced after rhGH therapy. Moreover, 10 patients showed low COR levels. In the IGHD group, COR levels also decreased, but remained within the normal range. Conclusion: During rhGH therapy, COR levels were reduced, particularly in patients with MPHD. HPA axis should be monitored during rhGH therapy.
Related JoVE Video
Gene Polymorphism of rs556621 but Not rs11984041 is Associated with the Risk of Large Artery Atherosclerotic Stroke in a Xinjiang Uyghur Population.
J Stroke Cerebrovasc Dis
PUBLISHED: 02-21-2014
Show Abstract
Hide Abstract
Stroke is one of the main causes of death and adult chronic disability. Recently, 2 independent genome-wide association studies reported that the genetic variants (rs556621 and rs11984041) are significantly associated with large artery atherosclerosis (LAA).
Related JoVE Video
Arcuate AgRP neurons mediate orexigenic and glucoregulatory actions of ghrelin.
Mol Metab
PUBLISHED: 02-01-2014
Show Abstract
Hide Abstract
The hormone ghrelin stimulates eating and helps maintain blood glucose upon caloric restriction. While previous studies have demonstrated that hypothalamic arcuate AgRP neurons are targets of ghrelin, the overall relevance of ghrelin signaling within intact AgRP neurons is unclear. Here, we tested the functional significance of ghrelin action on AgRP neurons using a new, tamoxifen-inducible AgRP-CreER(T2) transgenic mouse model that allows spatiotemporally-controlled re-expression of physiological levels of ghrelin receptors (GHSRs) specifically in AgRP neurons of adult GHSR-null mice that otherwise lack GHSR expression. AgRP neuron-selective GHSR re-expression partially restored the orexigenic response to administered ghrelin and fully restored the lowered blood glucose levels observed upon caloric restriction. The normalizing glucoregulatory effect of AgRP neuron-selective GHSR expression was linked to glucagon rises and hepatic gluconeogenesis induction. Thus, our data indicate that GHSR-containing AgRP neurons are not solely responsible for ghrelin's orexigenic effects but are sufficient to mediate ghrelin's effects on glycemia.
Related JoVE Video
Different ratios of bone marrow mesenchymal stem cells and chondrocytes used in tissue-engineered cartilage and its application for human ear-shaped substitutes in vitro.
Cells Tissues Organs (Print)
PUBLISHED: 02-01-2014
Show Abstract
Hide Abstract
The application of chondrocyte-based cartilage tissue engineering is limited because of the lack of autologous cartilage sources and chondrocyte dedifferentiation after in vitro expansion. Coculture of bone marrow mesenchymal stem cells (BMSCs) and chondrocytes has been a promising strategy for cartilage engineering as chondrocytes can provide a chondrogenic environment for BMSCs. However, there are no systematic comparison studies for engineered cartilage constructed using different mixing ratios of BMSCs and chondrocytes, and the most effective mixing ratio with the lowest number of chondrocytes is unknown. Here, we set a gradient of mixing ratios of BMSCs to chondrocytes for an in vitro coculture system and compared the shape retention and quality of the engineered cartilage using macroscopic and histological assays, glycosaminoglycan content assessment and immunohistochemical staining of type II collagen, biomechanical evaluation and hypertrophy-related gene expression analysis. The results showed that at least 30% chondrocytes were required to generate cartilage tissue with satisfactory shape and quality. Therefore, we preliminarily assessed the feasibility of engineering a human ear-shaped substitute using a coculture system with a 7:3 ratio of BMSCs to chondrocytes. After 8 weeks of in vitro culture, the precise architecture of the human ear-shaped construct was well maintained with the typical cartilaginous composition confirmed by histological assays.
Related JoVE Video
Domain I-IV of ?2-glycoprotein I inhibits advanced glycation end product-induced angiogenesis by down-regulating vascular endothelial growth factor 2 signaling.
Mol Med Rep
PUBLISHED: 01-31-2014
Show Abstract
Hide Abstract
Advanced glycation end products (AGEs) are a contributing factor in the angiogenesis that is characteristic of proliferative diabetic retinopathy. However, a previous study made a promising observation that domain I?IV of ?2?glycoprotein I (DI?IV) inhibits angiogenesis in human umbilical vein cells. The present study aimed to confirm the inhibition of AGE?induced angiogenesis in retinal endothelial cells by DI?IV and to investigate the potential underlying mechanisms. The RF/6A rhesus macaque choroid?retinal vascular endothelial cell line was cultured in vitro and treated with AGEs in the presence or absence of different concentrations of DI?IV. The proliferation, migration and tube formation of the RF/6A cells were evaluated using MTS assays, in vitro wound healing assays and in vitro Matrigel angiogenesis assays, respectively. The mRNA expression of vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR) 2, VEGFR 1 and receptor for AGE (RAGE) were quantified by reverse transcription quantitative polymerase chain reaction. The expression of VEGFR?1, VEGFR?2 and the activation of protein kinase B (Akt) and extracellular signal?regulated kinase (ERK) were also assessed by western blot analysis. The results indicated that AGEs promoted the migration, proliferation and tube formation of RF/6A cells in vitro (P<0.05), increased the expression of VEGF, VEGFR?2 and RAGE (P<0.05) and increased the phosphorylation of Akt and ERK (P<0.05). DI?IV inhibited the increase in VEGFR?2 mRNA and protein, but did not inhibit the increase in VEGF or RAGE mRNAs. These results led to the conclusion that DI?IV inhibited AGE?induced angiogenesis in the RF/6A cells, which was accompanied by a downregulation in the expression of VEGFR?2 and its downstream phosphatidylinosol 3?kinase/Akt and mitogen?activated protein kinase/ERK1/2 pathways. These findings provide further support towards the treatment of proliferative diabetic retinopathy by interventions that act via a mechanism similar to that of DI?IV.
Related JoVE Video
Role of STIM1 in survival and neural differentiation of mouse embryonic stem cells independent of Orai1-mediated Ca2+ entry.
Stem Cell Res
PUBLISHED: 01-16-2014
Show Abstract
Hide Abstract
Store-operated Ca(2+) entry (SOCE) is an important Ca(2+) influx pathway in non-excitable cells. STIM1, an ER Ca(2+) sensor, and Orai1, a plasma membrane Ca(2+) selective channel, are the two essential components of the Ca(2+) release activated channel (CRAC) responsible for SOCE activity. Here we explored the role of STIM1 and Orai1 in neural differentiation of mouse embryonic stem (ES) cells. We found that STIM1 and Orai1 were expressed and functionally active in ES cells, and expressions of STIM1 and Orai1 were dynamically regulated during neural differentiation of mouse ES cells. STIM1 knockdown inhibited the differentiation of mouse ES cells into neural progenitors, neurons, and astrocytes. In addition, STIM1 knockdown caused severe cell death and markedly suppressed the proliferation of neural progenitors. Surprisingly, Orai1 knockdown had little effect on neural differentiation of mouse ES cells, but the neurons derived from Orai1 knockdown ES cells, like those from STIM1 knockdown cells, had defective SOCE. Taken together, our data indicate that STIM1 is involved in both early neural differentiation of ES cells and survival of early differentiated ES cells independent of Orai1-mediated SOCE.
Related JoVE Video
The integrated production of microbial lipids and bio-SiO2 from rice husks by an organic electrolytes pretreatment technology.
Bioresour. Technol.
PUBLISHED: 01-09-2014
Show Abstract
Hide Abstract
In this study, a full dissolution behavior of rice husks (RHs) in ionic liquids-based organic electrolytes was achieved, and physicochemical effect of the dissolution pretreatment on the structures of RHs was elucidated. The physicochemical changes led to an enhanced subsequent enzymatic saccharification of RHs, and a total reducing sugars (TRSs) yield of 0.70gg(-1), and a glucose yield of 0.43gg(-1) were obtained. The hydrolysates could be used as carbon sources for the cultivation of Rhodosporidium toruloides Y4 for the production of microbial lipids with a satisfactory productivity of cell biomass (13.3gL(-1)) and lipid content (32.5%) after 100h cultivation. Further pyrolysis of the residuals after the enzymatic hydrolysis at 600°C for 3h resulted in new uniform, spherical silica powder materials with particle size around 150nm, and surface area of 179.3m(2)g(-1).
Related JoVE Video
Unique Epitopes Recognized by Monoclonal Antibodies against HP-PRRSV: Deep Understanding of Antigenic Structure and Virus-Antibody Interaction.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is a member of the genus Arterivirus within the family Arteriviridae. N and GP3 proteins are the immunodominance regions of the PRRSV viral proteins. To identify the B-cell linear antigenic epitopes within HP-PRRSV N and GP3 proteins, two monoclonal antibodies (mAbs) against N and GP3 proteins were generated and characterized, designated as 3D7 and 1F10 respectively. The mAb 3D7 recognized only HuN4-F112 not the corresponding virulent strain (HuN4-F5). It also recognized two other commercial vaccines (JXA1-R and TJM-F92), but not two other HP-PRRSV strains (HNZJJ-F1 and HLJMZ-F2). The B-cell epitope recognized by the mAb 3D7 was localized to N protein amino acids 7-33. Western blot showed that the only difference amino acid between HuN4-F112-N and HuN4-F5-N did not change the mAb 3D7 recognization to N protein. The epitope targeted by the mAb 1F10 was mapped by truncated proteins. We found a new epitope (68-76aa) can be recognized by the mAb. However, the epitope could not be recognized by the positive sera, suggesting the epitope could not induce antibody in pigs. These results should extend our understanding of the antigenic structure of the N protein and antigen-antibody reactions of the GP3 protein in different species.
Related JoVE Video
Lipid production from corn stover by the oleaginous yeast Cryptococcus curvatus.
Biotechnol Biofuels
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Microbial lipids produced from lignocellulosic biomass hold great promise for the biodiesel industry. These lipids usually consist of three major processes: pretreatment, enzymatic hydrolysis and lipid production. However, the conventional strategy of using biomass hydrolysates as the feedstock for lipid production suffers from low lipid coefficient and prohibitively high costs. More cost-effective and integrated processes are required to advance lignocellulosic biomass-based microbial lipid technology.
Related JoVE Video
Genetic Association Study of TNFAIP3, IFIH1, IRF5 Polymorphisms with Polymyositis/Dermatomyositis in Chinese Han Population.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
Single-nucleotide polymorphisms (SNPs) in the TNFAIP3, IFIH1, and IRF5 genes have been associated with several auto-inflammation diseases, while the susceptibility between these genes and idiopathic inflammatory myopathies (IIMs) were not reported. This study aimed to investigate whether TNFAIP3, IFIH1, and IRF5 gene polymorphisms confer susceptibility for the IIMs in Chinese Han population.
Related JoVE Video

What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.