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.
Expression of Attractin in male reproductive tract of human and mice and its correlation with male reproduction.
J. Huazhong Univ. Sci. Technol. Med. Sci.
PUBLISHED: 08-21-2014
Show Abstract
Hide Abstract
The expression of Attractin mRNA and protein in testis and semen of human and male mice was investigated. Human testis and semen samples were all collected from Reproductive Center of Renmin Hospital, Wuhan University in December, 2012. Testis samples were collected from 7 cases of obstructive azoospermias when they were subjected to diagnosed testis biopsy, and 30 normal human semen samples were obtained from those cases of semen analysis. Adult mice testis tissues were obtained from 10 2-month-old male BALB/c mice, and 60 male mice at different ages were classified into 10 groups (day 1, 5, 10, 15, 21, 28, 35, 42, 56, and 120 respectively, n=6 each). The expression of Attractin mRNA and protein in testis was detected by RT-PCR and Western blotting respectively. Human semen samples were centrifuged into sperm plasma (SP) and sperm extract (SE), and mice sperm samples were collected from the epididymis of 10 adult male BALB/c mice. Western blotting was used to determine the Attractin protein expression level. Attractin mRNA and protein were expressed in the testis of both patients with obstructive azoospermias and adult Bcl/B mice. Quantitative RT-PCR revealed that no Attractin mRNA was detectable in day 1 male BALB/c mice group. The Attractin mRNA and protein levels were low on the day 10, and increased with age until day 56. On the day 120, the expression levels of Attractin were decreased. As for human semen samples, Attractin protein was expressed in both SP and SE, but didn't exist in samples from the epididymis of male BALB/c mice. It was suggested that Attractin acted as a novel active substance and was involved in male reproduction in both human and BALB/c mice, but it exerted a different expression profile in different mammal species.
Related JoVE Video
A new potential risk factor in patients with erectile dysfunction and premature ejaculation: folate deficiency.
Asian J. Androl.
PUBLISHED: 08-02-2014
Show Abstract
Hide Abstract
We investigated serum folic acid (FA) levels in patients with erectile dysfunction (ED) and/or premature ejaculation (PE). Fasting serum samples were obtained from 42 patients with ED, 36 with PE, 25 ED patients with PE, and 30 healthy men; the mean intravaginal ejaculation latency time (IELT) was measured during a 4 weeks baseline period. Levels of sex hormones (follicle-stimulating hormone, luteinizing hormone, total testosterone), homocysteine (Hcys), and FA were measured using chemiluminescent immunoassays. The sexual functions of PE patients and normal control men were evaluated using the Chinese Index of Premature Ejaculation (CIPE). The abridged International Index of Erectile Function-5 (IIEF-5) questionnaire was used to gauge erectile quality for ED patients and for normal controls. Serum FA concentrations were lower in ED (7.61 ± 3.97 ng ml -1), PE (9.37 ± 3.40 ng ml -1), and ED/PE (8.84 ± 4.28 ng ml -1 ) patients than in healthy men (12.23 ± 5.76 ng ml -1 , P < 0.05). No significant differences in sex hormone levels were found between patients with sexual dysfunction and healthy controls (P > 0.05). There were positive correlations between serum FA concentrations and CIPE scores (r = 0.530, P < 0.01), IIEF-5 scores (r = 0.589, P < 0.01), and IELT (r = 0.445, P < 0.01); negative correlations with Hcys concentrations (r = -0.487, P < 0.01) were found in all participants. These findings showed a strong relationship between serum FA levels and sexual dysfunction, possibly due to an effect of FA on the metabolism of nitric oxide, Hcys, and 5-hydroxytryptamine.
Related JoVE Video
Downregulation of phosphoglycerate kinase 1 by shRNA sensitizes U251 xenografts to radiotherapy.
Oncol. Rep.
PUBLISHED: 07-23-2014
Show Abstract
Hide Abstract
Phosphoglycerate kinase 1 (PGK1) has been demonstrated to be involved in radioresistance. The present study was designed to investigate the effect of PGK1 on the radioresistance in vivo. U251 glioma cells were transfected with the short hairpin RNA (shRNA)-PGK1 and pcDNA3.1-PGK1 using Lipofectamine 2000. The radiosensitivity of U251 xenografts was observed by tumor growth curve following radiotherapy. Quantitative PCR, western blot analysis and immunohistochemistry were performed to evaluate PGK1 expression in the xenografts from the different tumor models. The expression of PGK1 was maximally inhibited in response to shRNA4 at 24 h after the transfection in vitro. Tumor growth of the U251 xenografts was significantly inhibited following treatment with shRNA-PGK1 and radiotherapy. The expression of PGK1 in vivo at the mRNA and protein levels was downregulated by the treatment of shRNA1 when compared to levels following treatment with shNC and PBS after radiotherapy. The results showed that suppression of PGK1 enhanced the radiosensitivity of U251 xenografts and suggest that PGK1 may serve as a useful target in the treatment of radioresistant glioma.
Related JoVE Video
Comprehensive analysis of cystatin family genes suggests their putative functions in sexual reproduction, embryogenesis, and seed formation.
J. Exp. Bot.
PUBLISHED: 07-04-2014
Show Abstract
Hide Abstract
Cystatins are tightly bound and reversible inhibitors of cysteine proteases in C1A and C13 peptidase families, which have been identified in several species and shown to function in vegetative development and response to biotic/abiotic stresses in plants. Recent work revealed their critical role in regulating programmed cell death during embryogenesis in tobacco and suggested their more comprehensive roles in the process of sexual plant reproduction, although little is known about cystatin family genes in the processes. Here, 10 cystatin family genes in Nicotiana tabacum were identified using an expressed sequence tag (EST)-based gene clone strategy. Analysis of their biochemical properties showed that nine of them have the potency to inhibit the activities of both commercial cathepsin L-like proteases and extracted cysteine proteases from seeds, but with different K i values depending on the types of proteases and the developmental stages of the seed tested. This suggests that cystatin-dependent cathepsin L-like proteolytic pathways are probably important for early seed development. Comprehensive expression profile analysis revealed that cystatin family genes showed manifold variations in their transcription levels in different plant cell types, including the sperm, egg, and zygote, especially in the embryo and seed at different developmental stages. More interestingly, intracellular localization analysis of each cystatin revealed that most members of cystatin families are recognized as secretory proteins with signal peptides that direct them to the endoplasmic reticulum. These results suggest their widespread roles in cell fate determination and cell-cell communication in the process of sexual reproduction, especially in gamete and embryo development, as well as in seed formation.
Related JoVE Video
Association between optic nerve sheath diameter and mortality in patients with severe traumatic brain injury.
Neurocrit Care
PUBLISHED: 06-28-2014
Show Abstract
Hide Abstract
Increased intracranial pressure (ICP) is associated with worse outcomes following traumatic brain injury (TBI). Studies have confirmed that ICP is correlated with optic nerve sheath diameter (ONSD) on ultrasound. The aim of our study was to assess the independent relationship between ONSD measured using CT and mortality in a population of patients admitted with severe TBI.
Related JoVE Video
Exogenous ?-aminobutyric acid (GABA) affects pollen tube growth via modulating putative Ca2+-permeable membrane channels and is coupled to negative regulation on glutamate decarboxylase.
J. Exp. Bot.
PUBLISHED: 05-05-2014
Show Abstract
Hide Abstract
?-Aminobutyric acid (GABA) is implicated in pollen tube growth, but the molecular and cellular mechanisms that it mediates are largely unknown. Here, it is shown that exogenous GABA modulates putative Ca(2+)-permeable channels on the plasma membranes of tobacco pollen grains and pollen tubes. Whole-cell voltage-clamp experiments and non-invasive micromeasurement technology (NMT) revealed that the influx of Ca(2+) increases in pollen tubes in response to exogenous GABA. It is also demonstrated that glutamate decarboxylase (GAD), the rate-limiting enzyme of GABA biosynthesis, is involved in feedback controls of Ca(2+)-permeable channels to fluctuate intracellular GABA levels and thus modulate pollen tube growth. The findings suggest that GAD activity linked with Ca(2+)-permeable channels relays an extracellular GABA signal and integrates multiple signal pathways to modulate tobacco pollen tube growth. Thus, the data explain how GABA mediates the communication between the style and the growing pollen tubes.
Related JoVE Video
Auxin transport and activity regulate stomatal patterning and development.
Nat Commun
PUBLISHED: 01-28-2014
Show Abstract
Hide Abstract
Stomata are two-celled valves that control epidermal pores whose spacing optimizes shoot-atmosphere gas exchange. They develop from protodermal cells after unequal divisions followed by an equal division and differentiation. The concentration of the hormone auxin, a master plant developmental regulator, is tightly controlled in time and space, but its role, if any, in stomatal formation is obscure. Here dynamic changes of auxin activity during stomatal development are monitored using auxin input (DII-VENUS) and output (DR5:VENUS) markers by time-lapse imaging. A decrease in auxin levels in the smaller daughter cell after unequal division presages the acquisition of a guard mother cell fate whose equal division produces the two guard cells. Thus, stomatal patterning requires auxin pathway control of stem cell compartment size, as well as auxin depletion that triggers a developmental switch from unequal to equal division.
Related JoVE Video
Effects of rapamycin on DC-SIGN expression and biological functions in DC.
Front Biosci (Landmark Ed)
PUBLISHED: 01-07-2014
Show Abstract
Hide Abstract
Rapamycin, a macrolide antibiotic, has potent immunosuppressive properties as an antirejection therapy in organ transplantation. Studies show that dendritic cells (DC) are important targets for rapamycin, which can inhibit DC maturation and DC-induced allogeneic T cell proliferation. In this study, we investigated the effects of rapamycin on the expressions of DC-SIGN and transcription factor PU.1 and the function of DC. Treatment with rapamycin significantly reduced the expression of DC-SIGN in a dose-dependent manner associated with suppression of PU.1 gene expression and the ability of DC to migrate and stimulate T cell proliferation. The expression of DC-SIGN was significantly suppressed using PU.1 siRNA. Intriguingly, rapamycin treatment largely decreased the expressions of PU.1 and DC-SIGN in THP-1 cells. In addition, treatment with rapamycin down-regulated the promoter activity of DC-SIGN. In conclusion, rapamycin inhibits DC-SIGN expression and suppresses the ability of DC to migrate and stimulate T cell proliferation through the PU.1 gene transcription pathway.
Related JoVE Video
Hypoxia Induces autophagic cell death through hypoxia-inducible factor 1? in microglia.
PLoS ONE
PUBLISHED: 01-01-2014
Show Abstract
Hide Abstract
As phagocytic cells of central nervous system, excessive activation or cell death of microglia is involved in a lot of nervous system injury and degenerative disease, such as stroke, epilepsy, Parkinson's disease, Alzheimer's disease. Accumulating evidence indicates that hypoxia upregulates HIF-1? expression leading to cell death of microglia. However, the exact mechanism of cell death induced by hypoxia in microglia is not clear. In the current study, we showed that hypoxia induced cell death and autophagy in microglia. The suppression of autophagy using either pharmacologic inhibitors (3-methyladenine, bafilomycin A1) or RNA interference in essential autophagy genes (BECN1 and ATG5) decreased the cell death induced by hypoxia in microglia cells. Moreover, the suppression of HIF-1? using either pharmacologic inhibitors (3-MA, Baf A1) or RNA interference decreased the microglia death and autophagy in vitro. Taken together, these data indicate that hypoxia contributes to autophagic cell death of microglia through HIF-1?, and provide novel therapeutic interventions for cerebral hypoxic diseases associated with microglia activation.
Related JoVE Video
Beneficial effects of the Chinese herbal medicine Sanjie Zhentong Capsule on experimental endometriosis in rats.
Chin J Nat Med
PUBLISHED: 12-19-2013
Show Abstract
Hide Abstract
To analyze the composition of the Chinese herbal medicine Sanjie Zhentong Capsule (SJZTC) and test the therapeutic efficacy of each component in a rat model of endometriosis.
Related JoVE Video
CD8(+) T cell-mediated cytotoxicity toward Schwann cells promotes diabetic peripheral neuropathy.
Cell. Physiol. Biochem.
PUBLISHED: 08-22-2013
Show Abstract
Hide Abstract
Damage to Schwann cells has been reported in the development of diabetic peripheral neuropathy (DPN), but how Schwann cells are damaged has not been elucidated.
Related JoVE Video
Daucosterol promotes the proliferation of neural stem cells.
J. Steroid Biochem. Mol. Biol.
PUBLISHED: 07-06-2013
Show Abstract
Hide Abstract
Neural stem cells (NSCs) are self-regenerating cells, but their regenerative capacity is limited. The present study was conducted to investigate the effect of daucosterol (a sterolin) on the promotion of NSC proliferation and determine the corresponding molecular mechanism. Results of cell counting kit-8 (CCK-8) assay showed that daucosterol significantly increased the quantity of viable cells and the effectiveness of daucosterol was similar to that of basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Flow cytometry detection of CFSE-labeled (CFSE, carboxyfluorescein diacetate succinimidyl ester) NSCs showed that Div Index (or the average number of cell divisions) and % Divided (or the percentage of cells that divided at least once) of the cells were increased, indicating that daucosterol increased the percentage of NSCs re-entering the cell cycle. mRNA microarray analysis showed that 333 genes that are mostly involved in the mitotic cell cycle were up-regulated. By contrast, 627 genes that are mostly involved in differentiation were down-regulated. In particular, insulin-like growth factor I (IGF1) was considered as an important regulatory gene that functionally promoted NSC proliferation, and the increased expression of IGF1 protein was validated by ELISA. In addition, the phosphorylation of AKT was increased, indicating that the proliferation-enhancing activity of daucosterol may be involved in IGF1-AKT pathway. Our study provided information about daucosterol as an efficient and inexpensive growth factor alternative that could be used in clinical medicine and research applications.
Related JoVE Video
Regulation of C-type natriuretic peptides and natriuretic peptide receptor-B expression in diabetic rats renal treated by Tongluo Recipe.
Chin J Integr Med
PUBLISHED: 07-02-2013
Show Abstract
Hide Abstract
To investigate the expression of C-type natriuretic peptides (CNP) and natriuretic peptide receptor-B (NPR-B) receptor in diabetic rats renal cortex, and the regulation by Tongluo Recipe (TLR).
Related JoVE Video
Heat shock factor OsHsfB2b negatively regulates drought and salt tolerance in rice.
Plant Cell Rep.
PUBLISHED: 06-22-2013
Show Abstract
Hide Abstract
Expression of OsHsfB2b was strongly induced by heat, salt, ABA and PEG treatments. Drought and salt tolerances were significantly decreased by OsHsfB2b overexpression, but were enhanced by RNA interference.
Related JoVE Video
Effect of lesion location on endometriotic adhesion and angiogenesis in SCID mice.
Arch. Gynecol. Obstet.
PUBLISHED: 06-05-2013
Show Abstract
Hide Abstract
To evaluate the effects of lesion location on adhesion and angiogenesis of transplanted endometriotic lesions in SCID mice.
Related JoVE Video
A differential role of macrophage TRPM2 channels in Ca²? signaling and cell death in early responses to H?O?.
Am. J. Physiol., Cell Physiol.
PUBLISHED: 04-17-2013
Show Abstract
Hide Abstract
Reactive oxygen species such as H?O? elevates the cytosolic Ca²? concentration ([Ca²?]c) and causes cell death via poly(ADPR) polymerase (PARP) activation, which also represents the primary mechanism by which H?O? activate the transient receptor potential melastatin-related 2 (TRPM2) channel as a Ca²?-permeable channel present in the plasma membrane or an intracellular Ca²?-release channel. The present study aimed to define the contribution and mechanisms of the TRPM2 channels in macrophage cells in mediating Ca²? signaling and cell death during initial response to H?O?, using mouse peritoneal macrophage, RAW264.7, and differentiated THP-1 cells. H?O? evoked robust increases in the [Ca²?]c, and such Ca²? responses were significantly greater at body temperature than room temperature. H?O?-induced Ca²? responses were strongly inhibited by pretreatment with PJ-34, a PARP inhibitor, and largely prevented by removal of extracellular Ca²?. Furthermore, H?O?-induced increases in the [Ca²?]c were completely abolished in macrophage cells isolated from trpm2-/- mice. H?O? reduced macrophage cell viability in a duration- and concentration-dependent manner. H?O?-induced cell death was significantly attenuated by pretreatment with PJ-34 and TRPM2 channel deficiency but remained significant and persistent. Taken together, these results show that the TRPM2 channel in macrophage cells functions as a cell surface Ca²?-permeable channel that mediates Ca²? influx and constitutes the principal Ca²? signaling mechanism but has a limited, albeit significant, role in cell death during early exposure to H?O?.
Related JoVE Video
Epstein Barr virus DNA analysis in blood predicts disease progression in a rare case of plasmablastic lymphoma with effusion.
Infect. Agents Cancer
PUBLISHED: 02-07-2013
Show Abstract
Hide Abstract
In HIV-1-infected patients a long lasting CD4+ cell decline influences the host-EBV balance and thereby increases the risk for EBV related malignancies. In spite of a world-wide access to combination antiretroviral therapy (cART) there are still a considerable number of HIV-1-infected patients who will develop severe immunodeficiency. These undiagnosed HIV-1 infected patients, so called late testers, demonstrate an increased lymphoma risk, compared to patients diagnosed early. Consecutive individual screening for EBV DNA-load in late testers might be a useful predictor of emerging EBV-malignancy.
Related JoVE Video
Over-expression of OsHsfA7 enhanced salt and drought tolerance in transgenic rice.
BMB Rep
PUBLISHED: 01-29-2013
Show Abstract
Hide Abstract
Heat shock proteins play an important role in plant stress tolerance and are mainly regulated by heat shock transcription factors (Hsfs). In this study, we generated transgenic rice over-expressing OsHsfA7 and carried out morphological observation and stress tolerance assays. Transgenic plants exhibited less, shorter lateral roots and root hair. Under salt treatment, over-expressing OsHsfA7 rice showed alleviative appearance of damage symptoms and higher survival rate, leaf electrical conductivity and malondialdehyde content of transgenic plants were lower than those of wild type plants. Meanwhile, transgenic rice seedlings restored normal growth but wild type plants could not be rescued after drought and re-watering treatment. These findings indicate that over-expression of OsHsfA7 gene can increase tolerance to salt and drought stresses in rice seedlings.
Related JoVE Video
Effect of oxygen tensions on the proliferation and angiogenesis of endometriosis heterograft in severe combined immunodeficiency mice.
Fertil. Steril.
PUBLISHED: 01-20-2013
Show Abstract
Hide Abstract
To investigate the effects of oxygen on the proliferation and angiogenesis of endometriosis in vivo.
Related JoVE Video
Notch1 is required for hypoxia-induced proliferation, invasion and chemoresistance of T-cell acute lymphoblastic leukemia cells.
J Hematol Oncol
PUBLISHED: 01-05-2013
Show Abstract
Hide Abstract
Notch1 is a potent regulator known to play an oncogenic role in many malignancies including T-cell acute lymphoblastic leukemia (T-ALL). Tumor hypoxia and increased hypoxia-inducible factor-1? (HIF-1?) activity can act as major stimuli for tumor aggressiveness and progression. Although hypoxia-mediated activation of the Notch1 pathway plays an important role in tumor cell survival and invasiveness, the interaction between HIF-1? and Notch1 has not yet been identified in T-ALL. This study was designed to investigate whether hypoxia activates Notch1 signalling through HIF-1? stabilization and to determine the contribution of hypoxia and HIF-1? to proliferation, invasion and chemoresistance in T-ALL.
Related JoVE Video
Outer membrane protein OmpW of Escherichia coli is required for resistance to phagocytosis.
Res. Microbiol.
PUBLISHED: 01-04-2013
Show Abstract
Hide Abstract
Eight-stranded ?-barrel outer membrane proteins can confer bacterial virulence via resistance to host innate defenses. This resistance function of OmpW, which was recently identified as an eight-stranded ?-barrel protein, was investigated in this study. Our results demonstrated that upregulation of OmpW correlated with increased bacterial survival during phagocytosis. Bacterial mutants harboring a deletion of ompW exhibited a significantly increased phagocytosis rate. Both observations suggest that the OmpW protein protects bacteria against host phagocytosis. In addition, expression of ompW is regulated by iron, which implies that the resistance provided by OmpW may be an important factor in iron-related infectious diseases. Furthermore, OmpW has been identified as a protective antigen that protects mice against bacterial infection and is therefore a promising target for vaccine development against infectious diseases.
Related JoVE Video
XRCC1 gene polymorphisms and the risk of differentiated thyroid carcinoma (DTC): a meta-analysis of case-control studies.
PLoS ONE
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
Previous studies investigating the association between X-ray repair cross-complementing group 1 (XRCC1) polymorphisms and thyroid cancer risk have yielded inconsistent results. This meta-analysis was performed to derive a more precise estimation of the relationship between three XRCC1 polymorphisms and thyroid cancer risk.
Related JoVE Video
Resting-state magnetoencephalography study of "small world" characteristics and cognitive dysfunction in patients with glioma.
Onco Targets Ther
PUBLISHED: 01-01-2013
Show Abstract
Hide Abstract
The purpose of this study was to analyze "small world" characteristics in glioma patients in order to understand the relationship between cognitive dysfunction and brain functional connectivity network in the resting state.
Related JoVE Video
[Survival and reproduction of Oncomelania hupensis robertsoni in water network regions in Hubei Province, China].
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi
PUBLISHED: 12-15-2011
Show Abstract
Hide Abstract
To understand the possibility of the survival and reproduction of Oncomelania hupensis robertsoni snails in water network regions in Hubei Province, as well as their expected lifespan after leaving the original propagating environment, so as to provide the basic data and precondition for reducing the infection rates of O. h. hupensis in the lake regions.
Related JoVE Video
[Gastroscopic and pathological characteristics of patients with or without schistosomiasis in Poyang Lake area].
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi
PUBLISHED: 12-15-2011
Show Abstract
Hide Abstract
To understand the gastroscopic and pathological characteristics of patients with miasis in Poyang Lake area, and to explore the relationship between schistosomiasis and pathological changes of gastric mucosa.
Related JoVE Video
Management with the insufficient proximal landing zone for endovascular repair in aortic dissection.
Chin. Med. J.
PUBLISHED: 11-02-2011
Show Abstract
Hide Abstract
Sufficient length of the proximal landing zone (PLZ) is the key for a successful thoracic endovascular aortic repair (TEVAR) of an aortic lesion. The aim of this research was to investigate the safety, feasibility, efficacy, and problems of endovascular repair for aortic dissection with insufficient PLZ.
Related JoVE Video
Association of two polymorphisms within and near SOCS3 gene with obesity in three nationalities in Xinjiang province of China.
Acta Pharmacol. Sin.
PUBLISHED: 09-05-2011
Show Abstract
Hide Abstract
SOCS3 gene plays an important role in the pathogenesis of obesity in animal models, but the data from human studies are relatively limited. To address this issue, a genetic association analysis on nationalities with different genetic background living in the similar environmental conditions was performed.
Related JoVE Video
A structural SVM approach for reference parsing.
BMC Bioinformatics
PUBLISHED: 06-09-2011
Show Abstract
Hide Abstract
Automated extraction of bibliographic data, such as article titles, author names, abstracts, and references is essential to the affordable creation of large citation databases. References, typically appearing at the end of journal articles, can also provide valuable information for extracting other bibliographic data. Therefore, parsing individual reference to extract author, title, journal, year, etc. is sometimes a necessary preprocessing step in building citation-indexing systems. The regular structure in references enables us to consider reference parsing a sequence learning problem and to study structural Support Vector Machine (structural SVM), a newly developed structured learning algorithm on parsing references.
Related JoVE Video
Serum biomarker of diabetic peripheral neuropathy indentified by differential proteomics.
Front Biosci (Landmark Ed)
PUBLISHED: 05-31-2011
Show Abstract
Hide Abstract
At least one in four diabetic patients is affected by peripheral neuropathy. In this study, the MALDI-TOF-MS mass spectra of peptides and proteins were generated following WCX CLINPROT bead fractionation of 39 diabetic peripheral neuropathy (DPN), 39 diabetes mellitus (DM), and 35 control (CON) serum samples. The spectra were analyzed statistically using flexAnalysisTM and Clin-ProtTM bioinformatics software. Identification of the selected markers was performed and affinity bead-purified plasma protein was subjected to LTQ Orbitrap XL MS/MS analysis followed by Mascot identification of the peptide sequences. 89 differentially expressed peaks of serum proteins were identified. 17, 10 and 4 most significant peaks between CON vs. DM, CON vs. DPN, DM vs. DPN, respectively, were selected out using the ClinProTool software package and used to train a Supervised Neural Network. A veracity rate of 100% was obtained for all sets. Following this analysis, a 6631-Da marker was identified as a fragment of the Apolipoprotein C-I precursor. The peptides identified may have clinical utility as surrogate markers for detection and classification of DM and DPN.
Related JoVE Video
Zinc inactivates melastatin transient receptor potential 2 channels via the outer pore.
J. Biol. Chem.
PUBLISHED: 05-20-2011
Show Abstract
Hide Abstract
Zinc ion (Zn(2+)) is an endogenous allosteric modulator that regulates the activity of a wide variety of ion channels in a reversible and concentration-dependent fashion. Here we used patch clamp recording to study the effects of Zn(2+) on the melastatin transient receptor potential 2 (TRPM2) channel. Zn(2+) inhibited the human (h) TRPM2 channel currents, and the steady-state inhibition was largely not reversed upon washout and concentration-independent in the range of 30-1000 ?M, suggesting that Zn(2+) induces channel inactivation. Zn(2+) inactivated the channels fully when they conducted inward currents, but only by half when they passed outward currents, indicating profound influence of the permeant ion on Zn(2+) inactivation. Alanine substitution scanning mutagenesis of 20 Zn(2+)-interacting candidate residues in the outer pore region of the hTRPM2 channel showed that mutation of Lys(952) in the extracellular end of the fifth transmembrane segment and Asp(1002) in the large turret strongly attenuated or abolished Zn(2+) inactivation, and mutation of several other residues dramatically changed the inactivation kinetics. The mouse (m) TRPM2 channels were also inactivated by Zn(2+), but the kinetics were remarkably slower. Reciprocal mutation of His(995) in the hTRPM2 channel and the equivalent Gln(992) in the mTRPM2 channel completely swapped the kinetics, but no such opposing effects resulted from exchanging another pair of species-specific residues, Arg(961)/Ser(958). We conclude from these results that Zn(2+) inactivates the TRPM2 channels and that residues in the outer pore are critical determinants of the inactivation.
Related JoVE Video
Proteomics of rice in response to heat stress and advances in genetic engineering for heat tolerance in rice.
Plant Cell Rep.
PUBLISHED: 04-08-2011
Show Abstract
Hide Abstract
Rice is the most important food crop worldwide. Global warming inevitably affects the grain yields of rice. Recent proteomics studies in rice have provided evidence for better understanding the mechanisms of thermal adaptation. Heat stress response in rice is complicated, involving up- or down-regulation of numerous proteins related to different metabolic pathways. The heat-responsive proteins mainly include protection proteins, proteins involved in protein biosynthesis, protein degradation, energy and carbohydrate metabolism, and redox homeostasis. In addition, increased thermotolerance in transgenic rice was obtained by overexpression of rice genes and genes from other plants. On the other hand, heterologous expression of some rice proteins led to enhanced thermotolerance in bacteria and other easily transformed plants. In this paper, we review the proteomic characterization of rice in response to high temperature and achievements of genetic engineering for heat tolerance in rice.
Related JoVE Video
Tumor necrosis factor-? increases angiopoietin-like protein 2 gene expression by activating Foxo1 in 3T3-L1 adipocytes.
Mol. Cell. Endocrinol.
PUBLISHED: 03-29-2011
Show Abstract
Hide Abstract
Angiopoietin-like protein 2 (Angptl2) is a key adipocyte-derived inflammatory mediator linking obesity to systemic insulin resistance, which is overexpressed in obesity and related metabolic diseases. However, its regulatory mechanism remains unclear. In this study, we showed that tumor necrosis factor (TNF)-? treatment increased the expression of Angptl2 gene in 3T3-L1 adipocytes. The cloning and sequence analysis of the Angptl2 gene promoter revealed the presence of several putative-binding sites for transcriptional factors, including two IREs. Insulin suppressed Angptl2 mRNA expression in dose-dependent manners, which could be attenuated by a phosphoinositide 3-kinase (PI3K) inhibitor LY294002. The interactions between IRE sites within Angptl2 promoter and forkhead transcription factor Foxo1 were identified by EMSA and ChIP assay. Furthermore, lentivirus-mediated knockdown of Foxo1 expression inhibited the transcriptional activity of Angptl2 promoter and decreased Angptl2 mRNA expression. Finally, TNF-? inhibited Foxo1 phosphorylation and enhanced its transcriptional activity, through which TNF-? increased the expression of Angptl2 in adipocytes. These results suggest that TNF-? up-regulates Angptl2 mRNA expression via PI3K/Foxo1 pathway in 3T3-L1 adipocytes, which may be involved in obesity-induced inflammation and insulin resistance.
Related JoVE Video
A residue in the TRPM2 channel outer pore is crucial in determining species-dependent sensitivity to extracellular acidic pH.
Pflugers Arch.
PUBLISHED: 02-16-2011
Show Abstract
Hide Abstract
Acidic pH is an important parameter regulating ion channel activity and its biological function. This study investigated inhibition of the hTRPM2 channels by extracellular acidic pH and compared the sensitivity of human (h) and mouse (m) TRPM2 channel to such an inhibition. The initial inhibition of hTRPM2 channel currents was substantially reversible, but the reversibility progressively diminished as the exposure to acidic pH was prolonged and it was essentially lost in the steady state, suggesting that extracellular acidic pH induces initial reversible inhibition and subsequent irreversible inactivation. Like the hTRPM2 channel, the mTRPM2 channel was sensitive to inhibition by pH 4.0-5.5, but the kinetics was significantly slower. Moreover, in contrast to the complete inhibition of the hTRPM2 channel, the mTRPM2 channel was insensitive to pH 6.0. Replacement of residue Gln(992) in the outer pore with the equivalent residue His(995) in the hTRPM2 channel resulted in a mutant mTRPM2 channel with the pH sensitivity and kinetics of inhibition of the wild-type hTRPM2 channel. Conversely, the reciprocal mutation H995Q in the hTRPM2 channel dramatically slowed down the kinetics of inhibition. Swapping other residues in the pore region failed to produce such opposing effects. Taken together, our results suggest a crucial role of residue His(995)/Gln(992) in the outer pore of TRPM2 channels in determining species-dependent effects of extracellular acidic pH.
Related JoVE Video
Core-sheath structured fibers with pDNA polyplex loadings for the optimal release profile and transfection efficiency as potential tissue engineering scaffolds.
Acta Biomater
PUBLISHED: 01-30-2011
Show Abstract
Hide Abstract
Emulsion electrospinning was initially applied to prepare core-sheath structured fibers with a core loading of pDNA or pDNA polyplexes inside a fiber sheath of poly(DL-lactide)-poly(ethylene glycol) (PELA). The inclusion of poly(ethylene imine) (PEI) and poly(ethylene glycol) (PEG) were expected to modulate the release profiles and achieve a balance between cytotoxicity and transfection efficiency. The core-sheath fibers enhance the structural integrity and maintain the biological activity of pDNA during the electrospinning process, incubation in release buffer and enzyme digestion. The addition of hydrophilic PEI into the fiber matrix accelerates pDNA release, while the encapsulation of pDNA polyplexes within the fibers led to no further release after an initial burst. However, sustained release of pDNA polyplexes has been achieved through PEG incorporation, and the effective release lifetime can be controlled between 6 and 25 days, dependent on the amount loaded and the molecular weight of PEG. Higher N/P ratios of PEI to DNA result in lower cell attachment, while cell viability is dependent on the effective concentration of pDNA polyplexes released from the fibers. While no apparent transfection is detected for pDNA-loaded PELA fibers, PEG incorporation into fibers containing pDNA polyplexes leads to over an order of magnitude increase in the transfection efficiency. pDNA polyplex-loaded fibers containing 10% PEG show the best performance in balancing transfection efficiency and cell viability. It is suggested that electrospun core-sheath fibers integrated with DNA condensation techniques provide the potential to produce inductive tissue engineering scaffolds able to manipulate the desired signals at effective levels within the local tissue microenvironment.
Related JoVE Video
Research on frequency of application with modern Chinese herbal medicine.
Chin J Integr Med
PUBLISHED: 01-22-2011
Show Abstract
Hide Abstract
To study the relationship between 500 kinds of commonly used Chinese herbal medicine and the classification of their efficacies in Chinese Materia Medica in relation to the common diseases listed in Internal Medicine.
Related JoVE Video
Effects of beraprost sodium, a prostaglandin I(2) analog, on high glucose-induced proliferation and oxidative stress in a rat glomerular mesangial cell line.
Pharmacology
PUBLISHED: 01-20-2011
Show Abstract
Hide Abstract
To investigate the effects of beraprost sodium on the proliferation and oxidative stress of glomerular mesangial cells under high glucose conditions, a rat mesangial cell line (rat mesangial cells; RMCs) was treated with beraprost sodium in the presence of high glucose concentrations. Proliferation rates of mesangial cells were detected by MTT assays and BrdU incorporation analyses. Levels of reactive oxygen species (ROS) were detected by DCFH-DA probes. The mRNA expression levels of CuZnSOD, MnSOD, catalase (CAT), glutathione peroxidase (Gpx), and collagen IV were detected by RT-PCR, and the protein levels of antioxidants (i.e. CuZnSOD, CAT, and MnSOD) and collagen IV were detected by Western blot. Beraprost sodium treatment significantly decreased the proliferation and ROS levels of RMCs cultured in high glucose conditions in a dose-dependent manner (p < 0.05). Beraprost sodium treatment decreased the mRNA and protein levels of CuZnSOD, CAT, and collagen IV in cells under high glucose conditions, while it increased MnSOD protein levels in cells under normal glucose conditions. Therefore, beraprost sodium inhibits high glucose-induced cellular proliferation and the generation of ROS, and it improves the antioxidant capacities of rat glomerular mesangial cells.
Related JoVE Video
Safety and immunogenicity of two freeze-dried Vero cell rabies vaccines for human use in post-exposure prophylaxis.
Vaccine
PUBLISHED: 01-06-2011
Show Abstract
Hide Abstract
To provide basis for human rabies vaccination in China, the safety and immunogenicity of two freeze-dried Vero cell rabies vaccines for human use were assessed. A total of 250 volunteers were enrolled and divided into two groups: volunteers in Group A (n=200) were vaccinated five doses of Speeda Vero cell rabies vaccine manufactured by Liaoning Chengda Biotechnology Co. Ltd. on day 0, 3, 7, 14, 28 after exposure. Volunteers in Group B (n=50) were treated with Verorab Vero cell rabies vaccine manufactured by Sanofi Pasteur on the same schedule. The local and systematic adverse reactions were observed. Serum neutralizing antibody levels of 80 individuals in Group A and 50 individuals in Group B were tested with RFFIT on day 7, 14, 45, 180, 360 after the first dose. The seroconversion rates in Groups A and B were 40.3% and 37.0% on day 7 after the first dose, 95.5% and 97.7% on day 14, 100% and 100% on day 45, 100% and 100% on day 180, 89.1% and 89.5% on day 360 respectively, indicating no significant differences between the two groups. And no significant differences were found between the neutralizing antibody geometric mean titers (GMTs) of the two groups on day 7, 14, 45, 180 and 360 after the first dose, with the GMTs of day 14, 45, 180 and 360 all higher than 0.5IU/ml. Antibody levels of the two groups peaked around 2 weeks after the full vaccination program, followed by a 55% decrease up to day 180 and another 76% decrease up to day 360. Both groups experienced occasions of transient fever, rash, edema, and scleroma after vaccination. Neither group had any severe adverse reactions. It was concluded that both vaccines showed satisfactory safety and immunogenicity. Booster vaccination is recommended following another exposure after six months since the full vaccination program.
Related JoVE Video
Residues 155 and 348 contribute to the determination of P2X7 receptor function via distinct mechanisms revealed by single-nucleotide polymorphisms.
J. Biol. Chem.
PUBLISHED: 01-04-2011
Show Abstract
Hide Abstract
P2X(7) receptors are important in mediating the physiological functions of extracellular ATP, and altered receptor expression and function have a causative role in the disease pathogenesis. Here, we investigated the mechanisms determining the P2X(7) receptor function by following two human single-nucleotide polymorphism (SNP) mutations that replace His-155 and Ala-348 in the human (h) P2X(7) receptor with the corresponding residues, Tyr-155 and Thr-348, in the rat (r) P2X(7) receptor. H155Y and A348T mutations in the hP2X(7) receptor increased ATP-induced currents, whereas the reciprocal mutations, Y155H and T348A, in the rP2X(7) receptor caused the opposite effects. Such a functional switch is a compelling indication that these residues are critical for P2X(7) receptor function. Additional mutations of His-155 and Ala-348 in the hP2X(7) receptor to residues with diverse side chains revealed a different dependence on the side chain properties, supporting the specificity of these two residues. Substitutions of the residues surrounding His-155 and Ala-348 in the hP2X(7) receptor with the equivalent ones in the rP2X(7) receptor also affected ATP-induced currents but were not fully reminiscent of the H155Y and A348T effects. Immunofluorescence imaging and biotin labeling assays showed that H155Y in the hP2X(7) receptor increased and Y155H in the rP2X(7) receptor decreased cell-surface expression. Such contrasting effects were not obvious with the reciprocal mutations of residue 348. Taken together, our results suggest that residues at positions 155 and 348 contribute to P2X(7) receptor function via determining the surface expression and the single-channel function, respectively. Such interpretations are consistent with the locations of the residues in the structural model of the hP2X(7) receptor.
Related JoVE Video
[Optical methods for in situ measuring leaf area index of forest canopy: a review].
Ying Yong Sheng Tai Xue Bao
PUBLISHED: 11-10-2010
Show Abstract
Hide Abstract
Leaf area index (LAI) is an important parameter of canopy structure, because it relates to many biophysical and physiological processes of canopy, including photosynthesis, respiration, transpiration, carbon cycling, precipitation interception, and energy exchange, etc. This paper introduced the theoretical bases and mathematical models of optical methods for forest canopy LAI determination, introduced the principles, merits, and drawbacks of currently used optical methods, and summed up the main sources of the errors in LAI optical measurement, including clumping effect, non-photosynthesis components, measurement conditions, and terrain effect. The developing status of quantitatively evaluating clumping effect, non-photosynthesis components, and terrain effect was analyzed, and the promising development directions of optical methods for measuring forest canopy LAI were discussed.
Related JoVE Video
Arabidopsis calcium-dependent protein kinase CPK10 functions in abscisic acid- and Ca2+-mediated stomatal regulation in response to drought stress.
Plant Physiol.
PUBLISHED: 08-30-2010
Show Abstract
Hide Abstract
Plant calcium-dependent protein kinases (CDPKs) may function as calcium sensors and play important roles in the regulation of plant growth and development and in plant responses to biotic and abiotic stresses. The Arabidopsis (Arabidopsis thaliana) genome encodes 34 CDPKs, and most of them have not been functionally characterized. Here, we report the functional characterization of CPK10 in Arabidopsis response to drought stress. The cpk10 mutant, a T-DNA insertion mutant for the Arabidopsis CPK10 gene, showed a much more sensitive phenotype to drought stress compared with wild-type plants, while the CPK10 overexpression lines displayed enhanced tolerance to drought stress. Induction of stomatal closure and inhibition of stomatal opening by abscisic acid (ABA) and Ca(2+) were impaired in the cpk10 mutants. Using yeast two-hybrid methods, a heat shock protein, HSP1, was identified as a CPK10-interacting protein. The interaction between CPK10 and HSP1 was further confirmed by pull-down and bimolecular fluorescence complementation assays. The HSP1 knockout mutant (hsp1) plants showed a similar sensitive phenotype under drought stress as the cpk10 mutant plants and were similarly less sensitive to ABA and Ca(2+) in regulation of stomatal movements. Electrophysiological experiments showed that ABA and Ca(2+) inhibition of the inward K(+) currents in stomatal guard cells were impaired in the cpk10 and hsp1 mutants. All presented data demonstrate that CPK10, possibly by interacting with HSP1, plays important roles in ABA- and Ca(2+)-mediated regulation of stomatal movements.
Related JoVE Video
MicroRNA-15a positively regulates insulin synthesis by inhibiting uncoupling protein-2 expression.
Diabetes Res. Clin. Pract.
PUBLISHED: 08-29-2010
Show Abstract
Hide Abstract
MicroRNAs are small noncoding RNAs that have been highly conserved during evolution and have been implicated to play an important role in many diseases, including diabetes. Several reports indicated the function of miRNAs in insulin production. However, the mechanisms by which miRNAs regulate this process remain poorly understood. Here we found that the expression of miR-15a was up-regulated in the presence of high glucose for 1h, whereas prolonged periods of high glucose exposure resulted in depressed expression of miR-15a, and the change in expression levels of miR-15a coincided with insulin biosynthesis. Moreover, ectopic expression of miR-15a promoted insulin biosynthesis in MIN6 cells, whereas its repression was sufficient to inhibit insulin biosynthesis. Further, we verified that miR-15a directly targeted and inhibited uncoupling protein-2 (UCP-2) gene expression. miR-15a mimics inhibited UCP-2 3UTR luciferase reporter activity. Western blot analysis showed that miR-15a inhibited endogenous UCP-2 protein levels, and resulted in the increase in oxygen consumption and reduced ATP generation. This study suggests miR-15a is a mediator of ? cell function and insulin biosynthesis, thus offering a new target for the development of preventive or therapeutic agents against diabetes.
Related JoVE Video
Mechanistic study of endogenous skin lesions in diabetic rats.
Exp. Dermatol.
PUBLISHED: 08-13-2010
Show Abstract
Hide Abstract
Pathological and physiological changes in dermal tissue in a rat model of diabetes mellitus (DM) were investigated. Sixteen male 8-week-old Sprague-Dawley rats were randomized into two groups of eight, the DM group (Group DM) and the normal control group (Group (NC) normal control). Group DM rats were injected with streptozotocin (STZ) intraperitoneally at a dose of 65 mg/kg body weight. Group NC rats were injected with the same volume of citric acid buffer. All rats were sacrificed 12 weeks later. The impact of exposure to (AGE) advanced glycation end products-modified human serum albumin (AGE-HSA) on epidermal cells and ECV304 cells was evaluated in cell culture experiments. The diabetic rats exhibited changes in skin tissue, including a decrease in thickness, disappearance of the multilayer epithelium structure, degeneration of collagen fibres and an increase in the infiltration of inflammatory cells, in addition to a significant increase in skin glucose and AGEs. Moreover, diabetic rats had increased plasma glycosylated protein (GSP) and malondialdehyde (MDA) and decreased plasma glutathione (GSH). The percentage of epidermal cells in S phase was similar between the two group rats; however, there was a marked decrease in the G2/M phase in Group DM. Additionally, exposure of ECV304 cells to AGE-HSA led to a time-dependent and dose-dependent increase in apoptosis. Therefore, the high glucose in the skin tissue, coupled with the accumulation of toxic substances such as AGEs, promote the dysfunction of dermal cells and/or the matrix. This may be a significant mechanism of diabetes-induced early-stage endogenous skin damage.
Related JoVE Video
TRPM2 channel properties, functions and therapeutic potentials.
Expert Opin. Ther. Targets
PUBLISHED: 07-31-2010
Show Abstract
Hide Abstract
Importance of the field: Oxidative stress, through production of reactive oxygen species, triggers disturbance in intracellular calcium [Ca(2+)](i) homeostasis, which has been identified as an important causative factor in the pathogenesis of numerous inflammatory, cardiovascular and neurodegenerative diseases. Areas covered in this review: Transient receptor potential melastatin 2 (TRPM2) protein forms a Ca(2+)-permeable cationic channel that is activated in response to oxidative stress and therefore acts as a cellular redox sensor. Research over the years has substantially advanced the knowledge of expression and functional properties of the TRPM2 channel, and particularly has accumulated compelling evidence for an important role for TRPM2 channel-mediated extracellular Ca(2+) influx in several physiological and pathophysiological functions exemplified by insulin release from pancreatic beta-cells, production of pro-inflammatory cytokines from immune cells, increased endothelial permeability, microglia activation and cell death. These findings suggest therapeutic potential of the TRPM2 channel as a drug target for combating oxidative-stress-related diseases. What the reader will gain: The current state of knowledge with respect to the TRPM2 channel properties and the roles in oxidant stress signalling and functions. Take home message: TRPM2 may be a novel therapeutic target for oxidative stress-related diseases.
Related JoVE Video
State-dependent inhibition of TRPM2 channel by acidic pH.
J. Biol. Chem.
PUBLISHED: 07-26-2010
Show Abstract
Hide Abstract
Transient receptor potential melastatin 2 (TRPM2) channel fulfills an important role in oxidative stress signaling in immune and other cells, to which local extracellular acidosis is known to occur under physiological or pathological conditions and impose significant effects on their functions. Here, we investigated whether the ADP-ribose-activated TRPM2 channel is a target for modulation by extracellular acidic pH by patch clamp recording of HEK293 cells expressing hTRPM2 channel. Induced whole cell or single channel currents were rapidly inhibited upon subsequent exposure to acidic pH. The inhibition in the steady state was complete, voltage-independent, and pH-independent in the range of pH 4.0-6.0. The inhibition was irreversible upon returning to pH 7.3, suggesting channel inactivation. In contrast, exposure of closed channels to acidic pH reduced the subsequent channel activation in a pH-dependent manner with an IC(50) for H(+) of 20 ?m (pH 4.7) and rendered subsequent current inhibition largely reversible, indicating differential or state-dependent inhibition and inactivation. Alanine substitution of residues in the outer vestibule of the pore including Lys(952) and Asp(1002) significantly slowed down or reduced acidic pH-induced inhibition and prevented inactivation. The results suggest that acidic pH acts as a negative feedback mechanism where protons bind to the outer vestibule of the TRPM2 channel pore and inhibit the TRPM2 channels in a state-dependent manner.
Related JoVE Video
Locating and parsing bibliographic references in HTML medical articles.
Int J Doc Anal Recognit
PUBLISHED: 07-20-2010
Show Abstract
Hide Abstract
The set of references that typically appear toward the end of journal articles is sometimes, though not always, a field in bibliographic (citation) databases. But even if references do not constitute such a field, they can be useful as a preprocessing step in the automated extraction of other bibliographic data from articles, as well as in computer-assisted indexing of articles. Automation in data extraction and indexing to minimize human labor is key to the affordable creation and maintenance of large bibliographic databases. Extracting the components of references, such as author names, article title, journal name, publication date and other entities, is therefore a valuable and sometimes necessary task. This paper describes a two-step process using statistical machine learning algorithms, to first locate the references in HTML medical articles and then to parse them. Reference locating identifies the reference section in an article and then decomposes it into individual references. We formulate this step as a two-class classification problem based on text and geometric features. An evaluation conducted on 500 articles drawn from 100 medical journals achieves near-perfect precision and recall rates for locating references. Reference parsing identifies the components of each reference. For this second step, we implement and compare two algorithms. One relies on sequence statistics and trains a Conditional Random Field. The other focuses on local feature statistics and trains a Support Vector Machine to classify each individual word, followed by a search algorithm that systematically corrects low confidence labels if the label sequence violates a set of predefined rules. The overall performance of these two reference-parsing algorithms is about the same: above 99% accuracy at the word level, and over 97% accuracy at the chunk level.
Related JoVE Video
[Effect of titanium nitride coating on bacterial corrosion resistance of dental Co-Cr alloy].
Shanghai Kou Qiang Yi Xue
PUBLISHED: 05-21-2010
Show Abstract
Hide Abstract
To study the influence of titanium nitride(TiN) coating on bacterial corrosion resistance of clinically used Co-Cr alloy.
Related JoVE Video
[Development of 3-enzyme pyrosequencing system and its application in rapid diagnosis of Downs syndrome].
Yi Chuan
PUBLISHED: 05-15-2010
Show Abstract
Hide Abstract
To avoid sequencing error resulting from use of apyrase in conventional 4- enzyme pyrosequencing system, a non-apyrase 3-enzyme pyrosequencing system with a better performance of quantitative analysis was established. The method is to immobilize biotinylated DNA template, ATP sulfurylase and luciferase on streptavidin-coated magnetic beads for pyrosequencing. After pyrosequencing, ATP produced from the pyrosequencing reaction and excess dNTPs were removed by magnetic separation technique; another dNTP was then dispensed for sequencing reaction, and the components interfering with the next circle of pyrosequencing reaction were removed by the same way, achieving the circular sequencing. This new system can accurately measure base sequences of a target DNA template, and also can quantitatively determine the relative ratio of two alleles. The allele ratios in two SNPs (rs1042917 and rs4818219) having a higher heterozygote rate on chromosome 21 were successfully detected for 16 normal samples and 8 clinical samples from Downs syndrome patients. The results can accurately demonstrate whether or not the target sample has equal copies of chromosome 21 from mother and father. This paper established a non-apyrase 3-enzyme pyrosequencing method, which owns a good perform-ance of quantitative analysis. The method is especially suitable to allelic quantification of an SNP, enabling the rapid diagnosis of Downs syndrome by analyzing allele ratio of SNPs on chromosome 21.
Related JoVE Video
Inhibition of ATP-induced macrophage death by emodin via antagonizing P2X7 receptor.
Eur. J. Pharmacol.
PUBLISHED: 03-19-2010
Show Abstract
Hide Abstract
Emodin (1,3,8-trihydroxy-6-methylanthraquinone), an anthraquinone derivative from Rheum officinale Baill, exhibits anti-inflammatory and immunosuppressive activities, however, the underlying mechanisms are not fully understood. This study examined the effects of emodin on ATP-evoked responses in rat peritoneal macrophages and in human embryonic kidney 293 cells (HEK293) heterologously expressing the cloned rat P2X7 receptor. Emodin reduced macrophage death induced by millimolar ATP in a concentration-dependent manner with the half of maximal inhibition values (IC50) of 0.2 microM. It also strongly inhibited ATP-induced dye uptake or pore formation, a hallmark property associated with P2X7 receptor activation, and 2,3-O-(benzoyl-4-benzoyl)-ATP (BzATP) induced increases in intracellular Ca2+ concentrations in macrophages with an IC50 of 0.5 microM. Furthermore, emodin significantly suppressed BzATP-evoked currents in P2X7 receptor expressing HEK293 cells with an IC50 of 3.4 microM. Taken together, these results provide compelling evidence for a novel action of emodin as a P2X7 receptor antagonist, which may underlie its anti-inflammatory and immunosuppressive activities.
Related JoVE Video
Release modulation and cytotoxicity of hydroxycamptothecin-loaded electrospun fibers with 2-hydroxypropyl-beta-cyclodextrin inoculations.
Int J Pharm
PUBLISHED: 01-04-2010
Show Abstract
Hide Abstract
Hydroxycamptothecin (HCPT) is valid to various malignant tumors, but its insoluble and unstable lactone ring in physiological environment have restricted the clinic application. This work was aimed to formulate HCPT-loaded poly(DL-lactic acid)-poly(ethylene glycol) (PELA) fibrous mats through blend electrospinning with 2-hydroxypropyl-beta-cyclodextrin (HPCD) to modulate the drug release and matrix degradation, and to enhance the structural integrity and cytotoxicity of the released HCPT. The entire drug fraction retained its active lactone form within electrospun fibers, and that was maintained over 85% during incubation for over 1 month. A biphasic release pattern was determined for HCPT-loaded electrospun fibers, which can be modulated by the addition of HPCD. HPCD served as solubilizer to maintain a large concentration gradient for HCPT between saturation and diffusion, and liberated HPCD created microstructure of ultrafine fibers, leading a faster release profile in the second phase. In vitro cytotoxicity test showed over 7 times higher inhibitory activity against cancer cells for HCPT-loaded electrospun fibers than free drug during 72h incubation. Higher apoptosis rates and the arrest of the cell cycle during the S and G(2)/M phases were detected through flow cytometry analysis. It indicated therapeutic potentials of HCPT-loaded electrospun fibers as implantable anti-cancer agents for local chemotherapy.
Related JoVE Video
[Studies on the chemical constituents of Dryopteris fragrans].
Zhong Yao Cai
PUBLISHED: 12-08-2009
Show Abstract
Hide Abstract
To study the chemical constituents of Dryopteris fragrans.
Related JoVE Video
[Genetic polymorphisms of fifteen short tandem repeats in Tujia ethnic group of Hubei province].
Zhonghua Yi Xue Yi Chuan Xue Za Zhi
PUBLISHED: 12-03-2009
Show Abstract
Hide Abstract
To investigate the genetic polymorphisms of 15 short tandem repeat (STR) loci in Tujia ethnic group of Hubei province, and to compare the allele frequencies between Hubei Tujia and Chongqing Tujia populations.
Related JoVE Video
18Beta-glycyrrhetinic acid ameliorates acute Propionibacterium acnes-induced liver injury through inhibition of macrophage inflammatory protein-1alpha.
J. Biol. Chem.
PUBLISHED: 11-06-2009
Show Abstract
Hide Abstract
18Beta-glycyrrhetinic acid (GA), the major bioactive component of licorice root extract, has a protective effect on hepatic injury and exhibits antiinflammatory activity. Here, we investigate the effect of GA in Propionibacterium acnes-induced acute inflammatory liver injury. C57BL/6 mice were primed with P. acnes followed by lipopolysaccharide challenge to induce fulminant hepatitis. GA (75 mg/kg) or vehicle control was administered intraperitoneally daily 1 day after P. acnes priming, and GA significantly improved mouse mortality. Then, to investigate the underlying mechanisms of GA in this acute inflammatory liver injury model, we primed C57BL/6 mice with P. acnes only. We propose that GA ameliorates acute P. acnes-induced liver injury through reduced macrophage inflammatory protein (MIP)-1alpha expression in Kupffer cells by down-regulating MyD88 expression and inhibiting NF-kappaB activation. Reduced MIP-1alpha expression lowered the recruitment of CD11c(+)B220(-) dendritic cell precursors into the liver. Consequently, GA treatment inhibits the activation and proliferation of liver-infiltrating CD4(+) T cells and reduces the production of serum alanine aminotransferase and proinflammatory cytokines such as interferon-gamma and tumor necrosis factor-alpha. Moreover, anti-MIP-1alpha treatment in P. acnes-primed mice inhibits the recruitment of dendritic cell precursors into the liver and suppresses mouse mortality as GA does. Taken together, our results suggest that GA exhibits antiinflammatory effects through inhibition of MIP-1alpha in a mouse model of acute P. acnes-induced inflammatory liver injury.
Related JoVE Video
[Dynamical distribution of bone marrow mesenchymal stem cells in rat model of chronic aristolochic acid nephropathy].
Zhongguo Zhong Xi Yi Jie He Za Zhi
PUBLISHED: 10-27-2009
Show Abstract
Hide Abstract
To investigate the distributive path and proliferative rule of marrow mesenchymal stem cells (MSCs) in the rat transplanted via caudal vein from male rat to female rats model of chronic aristolochic acid nephropathy (CAAN).
Related JoVE Video
Ion transporters involved in pollen germination and pollen tube tip-growth.
Plant Signal Behav
PUBLISHED: 09-10-2009
Show Abstract
Hide Abstract
Pollen germination (PG) and pollen tube growth (PTG) play crucial roles in sexual reproduction of flowering plants by sending sperm cells to the ovule. These two processes are regarded as ideal model system for the study of cell signaling and cell polarized growth. It has been considered for a long time that ion transports across the pollen tube membranes are essential for pollen tube navigation and growth. Previous transcriptome analyses for Arabidopsis have shown that the transcripts related to cellular transport are correspondingly overrepresented during the process of pollen tube growth. Here, we showed that 459 transporter genes expressed during PG and PTG in Arabidopsis. In addition, the gene expression profiles of ion (including Ca(2+), H(+), K(+), Cl(-)) channels and transporters were further analyzed. This analysis provides novel information for the potential candidate genes involving in ion fluxes across the pollen tube membranes and in regulation of pollen tube tip growth.
Related JoVE Video
Investigation of quinocetone-induced genotoxicity in HepG2 cells using the comet assay, cytokinesis-block micronucleus test and RAPD analysis.
Toxicol In Vitro
PUBLISHED: 07-16-2009
Show Abstract
Hide Abstract
Quinocetone, a new quinoxaline 1,4-dioxide derivative, has been approved as an animal growth promoter in China since 2003. To investigate the genotoxicity of quinocetone in vitro, its effects on the extent of DNA injury in human hepatoma (HepG2) cells accompanied by chromosomal damage and genomic DNA alterations were tested. The cell viability test indicated that quinocetone inhibited cell proliferation as a function of dose and time. In the comet assay, significant DNA fragment migration was observed in a dose-dependent manner. A dose-dependent increase of the micronucleated (MN) cell frequency was shown in cytokinesis-block micronucleus (CBMN) test. The gain/loss of randomly amplified polymorphic DNA (RAPD) bands and the change of band intensity in RAPD profiles were obtained after HepG2 cells were exposed to quinocetone at concentrations of 1.25, 2.5 and 5 microg/mL. The results demonstrated that quinocetone exerted genotoxic effects on HepG2 cells. Thus, the use of quinocetone as a growth promoter in animal feed should be seriously considered.
Related JoVE Video
Woody-to-total area ratio determination with a multispectral canopy imager.
Tree Physiol.
PUBLISHED: 06-24-2009
Show Abstract
Hide Abstract
Leaf area index (LAI) - defined as one half of the total green leaf area per unit ground surface area - can be determined by direct or indirect methods. Three major sources of errors exist in indirect LAI measurements: within-shoot clumping, beyond-shoot clumping and non-photosynthetic components. The effect of non-photosynthetic components on LAI measurements can be described by the woody-to-total area ratio, alpha; however, no convenient and efficient indirect methods have been developed to estimate alpha, especially the variations in alpha with zenith angle , alpha(theta). We describe the development and use of a multispectral canopy imager (MCI) to estimate alpha and alpha(theta) by considering the effects of non-random distributions of canopy elements and woody components and the overestimation of needle-to-shoot area ratio on woody components. The MCI, which mainly comprises a near-infrared band camera (Fujifilm IS-1), two visible band cameras (Canon 40D), filters and a pan tilt, was developed to measure clumping index, woody-to-total area ratio and geometric parameters of isolated trees. Two typical sampling plots (Plots 1 and 5) chosen from among 16 permanent forest experiment plots were selected for the estimation of alpha and alpha(theta). The non-random distributions of canopy elements and woody components were estimated separately at eight zenith angles (from 0 degrees to 70 degrees in increments of 10 degrees) using MCI images based on the gap size distribution theory. The visible/near-infrared image pairs captured by the MCI were able to discriminate among sky, leaves, cloud and woody components. Based on three methods of estimation, we obtained woody-to-total area ratios of 0.24, 0.19, 0.19 for Plot 1 and 0.23, 0.18, 0.17 for Plot 5. If clumping effects were ignored, alpha values were overestimated by as much as 21% and 24% at Plots 1 and 5, respectively. We demonstrated that alpha(theta) varied with the zenith angle, with variations in the range of 3-33% at Plot 1 and 2-65% at Plot 5. A new formula for the precise determination of LAI is proposed.
Related JoVE Video
[Quantitative pyrosequencing of heterozygous single nucleotide polymorphisms for rapid diagnosis of Downs syndrome].
Zhonghua Yi Xue Yi Chuan Xue Za Zhi
PUBLISHED: 06-09-2009
Show Abstract
Hide Abstract
To establish a method to detect Downs syndrome through quantitative pyrosequencing of the heterozygous single nucleotide polymorphisms (SNPs) on the chromosome 21.
Related JoVE Video
Effects of DC-SIGN expression on renal tubulointerstitial fibrosis in nephritis.
Front Biosci (Landmark Ed)
PUBLISHED: 03-11-2009
Show Abstract
Hide Abstract
Dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) is important for dendritic cell (DC) in migrating, recognizing, capturing, presenting antigens and in initiating T cell responses. In the present study, we investigated the role of DC-SIGN in renal tubulointerstitial inflammation and fibrosis. DC-SIGN was mainly expressed in tubular epithelial cells and DC-SIGN+ DCs were primarily distributed in renal tubulointerstitial areas during the early stage of nephritis, which was correlated with the degree of renal tubular interstitial lesions and fibrosis. In vitro, DC-SIGN expression in cultured human renal tubular epithelial cells was elevated when treated by tumor necrosis factor-alpha, and was inhibited by anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb). In a rat model of chronic renal interstitial fibrosis, there was a significant correlation of DC-SIGN expression with DC-SIGN+ DC distribution and the degree of tubulointerstitial lesion. PsL-EGFmAb reduced DC-SIGN expression and DC-SIGN+ DC accumulation in renal tissues in this rat model. These results suggest that DC-SIGN plays an important role in DC-mediated renal tubular interstitial lesions induced by immuno-inflammatory responses.
Related JoVE Video
Effects of DC-SIGN expression on renal tubulointerstitial fibrosis in nephritis.
Front Biosci (Landmark Ed)
PUBLISHED: 03-11-2009
Show Abstract
Hide Abstract
Dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN) is important for dendritic cell (DC) in migrating, recognizing, capturing, presenting antigens and in initiating T cell responses. In the present study, we investigated the role of DC-SIGN in renal tubulointerstitial inflammation and fibrosis. DC-SIGN was mainly expressed in tubular epithelial cells and DC-SIGN+ DCs were primarily distributed in renal tubulointerstitial areas during the early stage of nephritis, which was correlated with the degree of renal tubular interstitial lesions and fibrosis. In vitro, DC-SIGN expression in cultured human renal tubular epithelial cells was elevated when treated by tumor necrosis factor-alpha, and was inhibited by anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb). In a rat model of chronic renal interstitial fibrosis, there was a significant correlation of DC-SIGN expression with DC-SIGN+ DC distribution and the degree of tubulointerstitial lesion. PsL-EGFmAb reduced DC-SIGN expression and DC-SIGN+ DC accumulation in renal tissues in this rat model. These results suggest that DC-SIGN plays an important role in DC-mediated renal tubular interstitial lesions induced by immuno-inflammatory responses.
Related JoVE Video
Expression analysis of nine rice heat shock protein genes under abiotic stresses and ABA treatment.
J. Plant Physiol.
PUBLISHED: 01-08-2009
Show Abstract
Hide Abstract
Expression profiles of nine rice heat shock protein genes (OsHSPs) were analyzed by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The nine genes exhibited distinctive expression in different organs. Expression of nine OsHSP genes was affected differentially by abiotic stresses and abscisic acid (ABA). All nine OsHSP genes were induced strongly by heat shock treatment, whereas none of them were induced by cold. The transcripts of OsHSP80.2, OsHSP71.1 and OsHSP23.7 were increased during salt tress treatment. Expression of OsHSP80.2 and OsHSP24.1 genes were enhanced while treated with 10% PEG. Only OsHSP71.1 was induced by ABA while OsHSP24.1 was suppressed by ABA. These observations imply that the nine OsHSP genes may play different roles in plant development and abiotic stress responses.
Related JoVE Video
[Comparison of the effects of sanjie zhentong capsule and danazol on the endometriosis rats].
Zhongguo Zhong Xi Yi Jie He Za Zhi
Show Abstract
Hide Abstract
To compare the effects of sanjie zhentong capsule (SZC) and danazol on rats with endometriosis (EMT).
Related JoVE Video
Expression profile in rice panicle: insights into heat response mechanism at reproductive stage.
PLoS ONE
Show Abstract
Hide Abstract
Rice at reproductive stage is more sensitive to environmental changes, and little is known about the mechanism of heat response in rice panicle. Here, using rice microarray, we provided a time course gene expression profile of rice panicle at anther developmental stage 8 after 40°C treatment for 0 min, 20 min, 60 min, 2 h, 4 h, and 8 h. The identified differentially expressed genes were mainly involved in transcriptional regulation, transport, cellular homeostasis, and stress response. The predominant transcription factor gene families responsive to heat stress were Hsf, NAC, AP2/ERF, WRKY, MYB, and C(2)H(2). KMC analysis discovered the time-dependent gene expression pattern under heat stress. The motif co-occurrence analysis on the promoters of genes from an early up-regulated cluster showed the important roles of GCC box, HSE, ABRE, and CE3 in response to heat stress. The regulation model central to ROS combined with transcriptome and ROS quantification data in rice panicle indicated the great importance to maintain ROS balance and the existence of wide cross-talk in heat response. The present study increased our understanding of the heat response in rice panicle and provided good candidate genes for crop improvement.
Related JoVE Video
Effects of Chinese medicine Tongxinluo on hyperglycemia and beta-cell damage in streptozotocin-induced diabetic rats.
Chin. Med. J.
Show Abstract
Hide Abstract
Oxidative stress has been implicated in the onset and progression of diabetes. Tongxinluo is a traditional Chinese medicine with potent antioxidant properties. The aim of this study was to test the hypothesis that pretreatment with Tongxinluo has similar effects as melatonin on preventing hyperglycemia and beta-cell damage in a rat model of streptozotocin (STZ)-induced diabetes.
Related JoVE Video
[Research on detection of oil in water based on near-infrared scattering method].
Guang Pu Xue Yu Guang Pu Fen Xi
Show Abstract
Hide Abstract
The harm assessment and responsibility of marine mineral oil pollution require the content of mineral oil to be obtained by real-time online monitoring. In the present paper, the method of online monitoring of the oil content in water is presented, a creative technology based on optical scattering theory was proposed. Firstly the particle distribution of oil drops was scanned by Marvin laser particle meter, then with the help of the scattering signal, the crude, diesel of different concentrations from 0 to 30 mg x L(-1) was measured in experiment, the linearity is more than 0.99. This paper presents a feasible setup for the online monitoring of the content of mineral oil.
Related JoVE Video
[A review of researches on Angiostrongylus cantonensis and angiostrongyliasis cantonensis: analysis of related literatures].
Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi
Show Abstract
Hide Abstract
Angiostrongylus cantonensis is a lung worm of rat (definitive host). Human, an abnormal host, can acquire the infection by ingesting food containing the infective larvae of this worm (third-stage larvae). Although the larvae cannot develop into adult worm in human body, it moves to human brain and eyes, causing eosinophilic meningitis (EM), meningoencephalitis, or ocular angiostrongyliasis (OA). Here we review papers related to Angiostrongylus cantonensis and angiostrongyliasis cantonensis which were published in recent ten years, and identify the main institutes and principal investigators (PI) who are researching Angiostrongylus cantonensis. We also outline the advances in research on pathogen biology, pathogenesis, diagnostic techniques, epidemiology and prevention, as well as drug developed against Angiostrongylus cantonensis.
Related JoVE Video
Nitric oxide is involved in the regulation of trehalose accumulation under heat stress in Pleurotus eryngii var. tuoliensis.
Biotechnol. Lett.
Show Abstract
Hide Abstract
Little is known about the mechanism of how trehalose responds to various abiotic stresses although trehalose is considered as an important protectant in fungi. We investigated the role of nitric oxide (NO) in regulating trehalose accumulation during heat stress in Pleurotus eryngii var. tuoliensis. The addition of 100 or 200 g trehalose/l significantly inhibited the production of thiobarbituric acid-reactive substance under heat stress in mycelial cells. High temperature induced endogenous trehalose accumulation and sodium nitroprusside, a NO donor, further enhanced trehalose accumulation. Finally, heat-induced trehalose accumulation could be arrested by the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-1-oxyl-3-oxide, at 250 ?M by inhibiting the transcription of trehalose phosphate synthase gene. Thus NO plays an important role in the regulation of trehalose accumulation during abiotic stresses in P. eryngii var. tuoliensis.
Related JoVE Video
Over-expression of cofilin-1 and phosphoglycerate kinase 1 in astrocytomas involved in pathogenesis of radioresistance.
CNS Neurosci Ther
Show Abstract
Hide Abstract
Astrocytoma is among the most common intracranial tumors and radiotherapy is typically used after its resection. One of the outstanding problems encountered in the treatment is radioresistance. The lack of efficient biomarkers for evaluating the radiosensitivity of glioma precludes advances in treatment of astrocytoma and remains the most fatal cancer.
Related JoVE Video
Biodegradation of crude oil using an efficient microbial consortium in a simulated marine environment.
Mar. Pollut. Bull.
Show Abstract
Hide Abstract
Ochrobactrum sp. N1, Brevibacillus parabrevis N2, B. parabrevis N3 and B. parabrevis N4 were selected when preparing a mixed bacterial consortium based on the efficiency of crude oil utilization. A crude oil degradation rate of the N-series microbial consortium reached upwards of 79% at a temperature of 25 °C in a 3.0% NaCl solution in the shake flask trial. In the mesocosm experiment, a specially designed device was used to simulate the marine environment. The internal tank size was 1.5 m (L)×0.8 m (W)×0.7 m (H). The microbial growth conditions, nutrient utilization and environmental factors were thoroughly investigated. Over 51.1% of the crude oil was effectively removed from the simulated water body. The escalation process (from flask trials to the mesocosm experiment), which sought to represent removal under conditions more similar to the field, proved the high efficiency of using N-series bacteria in crude oil degradation.
Related JoVE Video
Pinching spine: A potential treatment for depression.
Chin J Integr Med
Show Abstract
Hide Abstract
ABSTACT: OBJECTIVE: To investigate whether pinching spine (PS, i.e., a traditional Chinese manipulative therapy) is benefificial to ameliorating the depressive state (including behavioral defificit, retardative weight gain and decreased sucrose consumption) in a rat model of depression induced by chronic unpredictable stress (CUS) and to explore the candidate mechanism of action. METHODS: PS was performed on rats spine once daily for 1 week after exposure to CUS. The open-fifield test, body weight measuring, and sucrose intake test were applied on different dates: before stress (d0), at the end of stress (d21) and after PS treatment (d28), respectively. Then the rats hippocampuses were performed genome-wide microarray analysis, and the expression levels of several genes were evaluated by real-time polymerase chain reaction (PCR). RESULTS: Exposure to CUS resulted in decreases of behavioral activity and sucrose consumption, which were reversed signifificantly after PS treatment. The expression of several genes relevant to energy metabolism, anti-oxidation, and olfactory receptor, etc., were down-regulated, while the expression of those relevant to hemostasis, immunity-inflammation, and restriction of activities and ingestion, etc., were up-regulated in hippocampuses of rats exposed to CUS. PS treatment signifificantly inverted these changes. Furthermore, increase or decrease in gene expression evaluated by realtime PCR was concordant with up-regulated or down-regulated expression evaluated by microarray analysis. CONCLUSION: PS showed a potential antidepressant-like effect, of which the action mechanism might be due to gene expression regulation in hippocampus.
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.