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Find video protocols related to scientific articles indexed in Pubmed.
[Determination of sodium, magnesium, calcium, lithium and strontium in natural mineral drinking water by microwave plasma torch spectrometer with nebulization sample introduction system].
Guang Pu Xue Yu Guang Pu Fen Xi
PUBLISHED: 11-01-2014
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The microwave plasma torch (MPT) was used as the emission light source. Aqueous samples were introduced with a nebulizer and a desolvation system. A method for the determination of Na, Mg, Ca, Li and Sr in natural mineral drinking water by argon microwave plasma torch spectrometer (ArMPT spectrometer) was established. The effects of microwave power, flow rate of carrier gas and support gas were investigated in detail and these parameters were optimized. Under the optimized condition, the experiments for the determination of Na, Mg, Ca, Li and Sr in 11 kinds of bottled mineral drinking water were carried out by ArMPT spectrometer. The limit-of-detection (LOD) of Na, Mg, Ca, Li and Sr was found to be 4.4, 21, 56, 11 and 84 ?g x mL(-1), respectively. Relative standard deviation (n = 6) was in the range of 1.30%-5.45% and standard addition recoveries were in the range of 84.6%-98.5%. MPT spectrometer was simpler, more convenient and of lower cost as compared to ICP unit. MPT spectrometer demonstrated its rapid analysis speed, accuracy, sensitivity and simultaneous multi element analysis ability during the analysis process. The results showed that MPT spectrometer was suitable for metal elements detection for natural mineral drinking water. This approach provides not only one way for resisting the illegal dealings, but also a security for the quality of drinking water. Moreover, the usability of MPT spectrometer in the field of food security; drug safety; clinical diagnostic is promised.
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Ordered arrays of a defect-modified ferroelectric polymer for non-volatile memory with minimized energy consumption.
Nanoscale
PUBLISHED: 10-16-2014
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Ferroelectric polymers are among the most promising materials for flexible electronic devices. Highly ordered arrays of the defect-modified ferroelectric polymer P(VDF-TrFE-CFE) (poly(vinylidene fluoride-trifluoroethylene-chlorofluoroethylene)) are fabricated by nanoimprint lithography for nonvolatile memory application. The defective CFE units reduce the coercive field to one-fifth of that of the un-modified P(VDF-TrFE), which can help minimize the energy consumption and extend the lifespan of the device. The nanoimprint process leads to preferable orientation of polymer chains and delicately controlled distribution of the defects, and thus a bi-stable polarization that makes the memory nonvolatile, as revealed by the pulsed polarization experiment.
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Rpb3 Promotes Hepatocellular Carcinoma through its N-terminus.
Oncotarget
PUBLISHED: 09-12-2014
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The expression of RNA polymerase II subunit 3 (Rpb3) was found frequent up-regulation in Hepatocellular carcinoma (HCC) tumors. Significant associations could also be drawn between increased expressions of Rpb3 and advance HCC staging and shorter disease-free survival of patients. Overexpression of Rpb3 increased HCC cell proliferation, migratory rate and tumor growth in nude mice, whereas suppression of Rpb3 using shRNA inhibited these effects. For mechanism study, we found that Rpb3 bound directly to Snail, downregulated E-cadherin, induced HCC cells epithelial-mesenchymal transition (EMT). In particular, N-terminus of Rpb3 blocked Rpb3 binding to Snail, inhibited Rpb3-high-expression HCC cells proliferation, migration, tumor growth in nude mice, and also inhibited DEN-induced liver tumorigenesis. Furthermore, N-terminus of Rpb3 did not inhibit normal liver cells or Rpb3-low-expression HCC cells proliferation. These findings suggest that N-terminus of Rpb3 selectively inhibits Rpb3-high-expression HCC cells proliferation. N-terminus of Rpb3 may be useful in treating patients diagnosed with Rpb3-high-expression HCC.
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IL-12 regulates B7-H1 expression in ovarian cancer-associated macrophages by effects on NF-?B signalling.
Asian Pac. J. Cancer Prev.
PUBLISHED: 08-02-2014
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B7-H1, a co-inhibitory molecule of the B7 family, is found aberrantly expressed in ovarian cancer cells and infiltrating macrophage/dendritic-like cells, and plays a critical role in immune evasion by ovarian cancer. IL-12, an inducer of Th1 cell development, exerts immunomodulatory effects on ovarian cancer. However, whether IL-12 regulates B7-H1 expression in human ovarian cancer associated-macrophages has not been clarified. Therefore, we investigated the effects of IL-12 on the expression of B7-H1 in ovarian cancer-associated macrophages and possible mechanisms.
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[Correlative analysis on metatarsalgia and the X-ray measurement indexes under weight-bearing and non-weight-bearing of hallux valgus].
Zhongguo Gu Shang
PUBLISHED: 07-18-2014
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To study changes in the radiographic appearance during weight-bearing and non-weigh-bearing in hallux valgus, and to analyse the correlation between the elasticity of plantar soft tissue of hallux valgus and the pain under the metatarsal head.
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The epidemic characteristics and changing trend of hemorrhagic fever with renal syndrome in Hubei Province, China.
PLoS ONE
PUBLISHED: 01-01-2014
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Hemorrhagic fever with renal syndrome (HFRS) is caused by different hantaviruses within the Bunyaviridae family. HFRS is a fulminant, infectious disease that occurs worldwide and is endemic in all 31 provinces of China. Since the first HFRS case in Hubei Province was reported in 1957, the disease has spread across the province and Hubei has become one of the seriously affected areas in China with the greatest number of reported HFRS cases in the 1980's. However, the epidemic characteristics of HFRS in Hubei are still not entirely clear and long-term, systematic investigations of this epidemic area have been very limited.
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In vitro and in vivo studies of the inhibitory effects of emodin isolated from Polygonum cuspidatum on Coxsakievirus B?.
Molecules
PUBLISHED: 07-29-2013
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The lack of effective therapeutics for Coxsackievirus B? (CVB?) infection underscores the importance of finding novel antiviral compounds. Emodin (1,3,8-trihydroxy-6-methylanthraquinone) is one of the natural anthraquinone derivatives obtained from the root and rhizome of Polygonum cuspidatum. In the present study, the possibility of using emodin as a potential antiviral to treat CVB? infection was explored in vitro and in mice. Emodin reduced CVB? entry and replication on Hep-2 cells in a concentration- and time-dependent manner, with a 50% effective concentration (EC??) of 12.06 ?M and selectivity index (SI) of 5.08, respectively. The inhibitory effect of emodin for CVB? entry and replication was further confirmed by a quantitative real time PCR (qPCR) assay. The results further showed that the mice orally treated with different dosages of emodin displayed a dose dependent increase of survival rate, body weight and prolonged mean time of death (MTD), accompanied by significantly decreased myocardial virus titers and pathologic scores/lesions. Moreover, emodin could inhibit CVB?-induced apoptosis in vitro and in vivo. Our results indicated that emodin could be used as potential antiviral in the post-exposure prophylaxis for CVB? infection.
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Alleviation of hyperglycemia induced vascular endothelial injury by exenatide might be related to the reduction of nitrooxidative stress.
Biomed Res Int
PUBLISHED: 07-07-2013
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We will investigate the effects of exenatide on vascular endothelial injury and nitrooxidative stress in hyperglycemia both in vivo and in vitro and explore the role of nitrooxidative stress in endothelium-protective action of exenatide. Healthy male Wistar rats were randomly divided into 4 groups: control, diabetes mellitus (DM) model, low dose of exenatide treatment, and high dose of exenatide treatment. In vitro study showed that, compared with control group, the DM rats exhibited a lowered endothelium-dependent relaxation and damaged structural integrity of thoracic aortas, and there was a significant increase in plasma nitrotyrosine concentration. These parameters were improved after treatment with either low dose or high dose of exenatide for 45 days. In vitro study showed that exendin-4 (the active ingredient of exenatide) attenuated HUVECs injury induced by high glucose, with improving cell viability and attenuating cell apoptosis. Exendin-4 also significantly alleviated the increased malondialdehyde (MDA), nitrotyrosine content, and inducible nitric oxide synthase (iNOS) expression induced by high glucose in HUVECs. In conclusion, this study demonstrates that exenatide treatment can alleviate the vascular endothelial injury, as well as attenuating the nitrooxidative stress in hyperglycemia, implying that the endothelium-protective effect of exenatide might be related to the reduction of nitrooxidative stress.
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Aberrant DNA methylation of P16, MGMT, hMLH1 and hMSH2 genes in combination with the MTHFR C677T genetic polymorphism in gastric cancer.
Asian Pac. J. Cancer Prev.
PUBLISHED: 06-28-2013
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Associations of P16, MGMT, hMLH1 and hMLH2 with gastric cancer and their relation with MTHFR status in gastric patients who were confirmed with pathological diagnosis were assessed. Aberrant DNA methylation of P16, MGMT, hMLH1 and hMLH2 and polymorphisms of MTHFR C677T were assayed. The proportional DNA hypermethylation in P16, MGMT, hMLH1 and hMLH2 in cancer tissues was significantly higher than in remote normal-appearing tissues. DNA hypermethylation of P16 and MGMT was correlated with the T and N stages. Individuals with homozygotes (TT) of MTHFR C677T had significant risk of hypermethylation of MGMT in cancer tissues [OR (95% CI)= 3.47(1.41-7.93)]. However, we did not find association between polymorphism in MTHFR C677T and risk of hypermethylation in P16, MGMT, hMLH1 and hMLH2 genes either in cancer or remote normal-appearing tissues. Aberrant hypermethylation of P16, MGMT, hMLH1 and hMLH2 could be predictive of gastric cancer.
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Enhanced viability of Lactobacillus reuteri for probiotics production in mixed solid-state fermentation in the presence of Bacillus subtilis.
Folia Microbiol. (Praha)
PUBLISHED: 06-07-2013
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In order to develop a multi-microbe probiotic preparation of Lactobacillus reuteri G8-5 and Bacillus subtilis MA139 in solid-state fermentation, a series of parameters were optimized sequentially in shake flask culture. The effect of supplementation of B. subtilis MA139 as starters on the viability of L. reuteri G8-5 was also explored. The results showed that the optimized process was as follows: water content, 50 %; initial pH of diluted molasses, 6.5; inocula volume, 2 %; flask dry contents, 30?35 g/250 g without sterilization; and fermentation time, 2 days. The multi-microbial preparations finally provided the maximum concentration of Lactobacillus of about 9.01?±?0.15 log CFU/g and spores of Bacillus of about 10.30?±?0.08 log CFU/g. Compared with pure fermentation of L. reuteri G8-5, significantly high viable cells, low value of pH, and reducing sugar in solid substrates were achieved in mixed fermentation in the presence of B. subtilis MA139 (P?
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Photosensitizing effects of nanometer TiO2 on chlorothalonil photodegradation in aqueous solution and on the surface of pepper.
J. Agric. Food Chem.
PUBLISHED: 05-17-2013
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The present study examined the effects of anatase nanometer TiO2 on photochemical degradation of chlorothalonil in aqueous solution and on the plant surface. Results showed that nanometer TiO2 exhibited a strong photosensitizing effect on the degradation of chlorothalonil both in aqueous solution and on the surface of green pepper. The photosensitization rate was the highest in the sunlight compared to illumination under high-pressure mercury and UV lamps. Use of distinct hydroxyl radical scavengers indicated that nanometer TiO2 acted by producing hydroxyl radicals with strong oxidizing capacity. Notably, nanometer TiO2 facilitated complete photodegradation of chlorothalonil with no detectable accumulation of the intermediate chlorothalonil-4-hydroxy. Nanometer TiO2 was also active on the surface of green pepper under natural sunlight both inside and outside of plastic greenhouse. These results together suggest that nanometer TiO2 can be used as a photosensitizer to accelerate degradation of the pesticides under greenhouse conditions.
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Proteomics-based identification of tumor relevant proteins in lung adenocarcinoma.
Biomed. Pharmacother.
PUBLISHED: 04-25-2013
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Lung cancer has the highest mortality rate among malignant tumors. Proteomics is a powerful tool to identify protein biomarkers. The identification of protein biomarkers associated with lung adenocarcinoma would have significance for making prognoses and designing targeted therapies.
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Antiviral and anti-inflammatory activity of arbidol hydrochloride in influenza A (H1N1) virus infection.
Acta Pharmacol. Sin.
PUBLISHED: 04-10-2013
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To investigate the effects of arbidol hydrochloride (ARB), a widely used antiviral agent, on the inflammation induced by influenza virus.
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Angiotensin II Receptor Blocker Telmisartan Prevents New-Onset Diabetes in Pre-Diabetes OLETF Rats on a High-Fat Diet: Evidence of Anti-Diabetes Action.
Can J Diabetes
PUBLISHED: 03-06-2013
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This study aims to investigate the effects of telmisartan, pioglitazone and metformin administration on the prevention of new-onset type 2 diabetes mellitus in pre-diabetes Otsuka Long-Evans Tokushima Fatty (OLETF) rats fed with a high-fat diet (HFD).
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Serum agonistic autoantibodies against type-1 angiotensin II receptor titer in patients with epithelial ovarian cancer: a potential role in tumor cell migration and angiogenesis.
J Ovarian Res
PUBLISHED: 03-03-2013
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Although agonistic autoantibodies against type-1 angiotensin-II receptor (AT1-AA) are frequently detected in women with preeclampsia, the clinical significance of AT1-AA in association with epithelial ovarian cancer (EOC) has not been identified.
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7-Deazapurine and 8-aza-7-deazapurine nucleoside and oligonucleotide pyrene "click" conjugates: synthesis, nucleobase controlled fluorescence quenching, and duplex stability.
J. Org. Chem.
PUBLISHED: 12-14-2011
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7-Deazapurine and 8-aza-7-deazapurine nucleosides related to dA and dG bearing 7-octadiynyl or 7-tripropargylamine side chains as well as corresponding oligonucleotides were synthesized. "Click" conjugation with 1-azidomethyl pyrene (10) resulted in fluorescent derivatives. Octadiynyl conjugates show only monomer fluorescence, while the proximal alignment of pyrene residues in the tripropargylamine derivatives causes excimer emission. 8-Aza-7-deazapurine pyrene "click" conjugates exhibit fluorescence emission much higher than that of 7-deazapurine derivatives. They are quenched by intramolecular charge transfer between the nucleobase and the dye. Oligonucleotide single strands decorated with two "double clicked" pyrenes show weak or no excimer fluorescence. However, when duplexes carry proximal pyrenes in complementary strands, strong excimer fluorescence is observed. A single replacement of a canonical nucleoside by a pyrene conjugate stabilizes the duplex substantially, most likely by stacking interactions: 6-12 °C for duplexes with a modified "adenine" base and 2-6 °C for a modified "guanine" base. The favorable photophysical properties of 8-aza-7-deazapurine pyrene conjugates improve the utility of pyrene fluorescence reporters in oligonucleotide sensing as these nucleoside conjugates are not affected by nucleobase induced quenching.
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Over-expression of Slit2 induces vessel formation and changes blood vessel permeability in mouse brain.
Acta Pharmacol. Sin.
PUBLISHED: 10-10-2011
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To investigate the effect of the axon guidance cue Slit2 on the density of blood vessels and permeability of the blood-brain barrier in mouse brain.
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Genetic analysis of hantaviruses and their rodent hosts in central-south China.
Virus Res.
PUBLISHED: 08-26-2011
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Hantaan virus (HTNV) and Seoul virus (SEOV) are two major zoonotic pathogens of hemorrhagic fever with renal syndrome (HFRS) in Asia. Hubei province, which is located in the central-south China, had been one of the most severe epidemic areas of HFRS. To investigate phylogenetic relationships, genetic diversity and geographic distribution of HTNV and SEOV in their reservoir hosts, a total of 687 rodents were trapped in this area between 2000 and 2009. Sequences of partial S- and M-segments of hantaviruses and mitochondrial D-loop gene from 30 positive samples were determined. Our data indicated that SEOV and HTNV were co-circulating in Hubei. Phylogenetic analysis based on partial S- and M-segment sequences revealed two and three previously undefined lineages of SEOV, and a novel genetic lineage of HTNV, respectively. Four inter-lineage reassortment SEOVs carried by Rattus norvegicus and Apodemus agrarius were observed. It suggests that SEOV may cause spillover infections to A. agrarius naturally. The abundance of the phylogenetic lineages of SEOV suggested that central-south China was a radiation center for SEOVs.
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Stepwise "click" chemistry for the template independent construction of a broad variety of cross-linked oligonucleotides: influence of linker length, position, and linking number on DNA duplex stability.
J. Org. Chem.
PUBLISHED: 06-16-2011
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Cross-linked DNA was constructed by a "stepwise click" reaction using a bis-azide. The reaction is performed in the absence of a template, and a monofunctionalized oligonucleotide bearing an azido-function is formed as intermediate. For this, an excess of the bis-azide has to be used compared to the alkynylated oligonucleotide. The cross-linking can be carried out with any alkynylated DNA having a terminal triple bond at any position of the oligonucleotide, independent of chain length or sequence with identical or nonidentical chains. Short and long linkers with terminal triple bonds were introduced in the 7-position of 8-aza-7-deaza-2-deoxyguanosine (1 or 2), and the outcome of the "stepwise" click and the "bis-click" reaction was compared. The cross-linked DNAs form cross-linked duplexes when hybridized with single-stranded complementary oligonucleotides. The stability of these cross-linked duplexes is as high as respective individual duplexes when they were ligated at terminal positions with linkers of sufficient length. The stability decreases when the linkers are incorporated at central positions. The highest duplex stability was reached when two complementary cross-linked oligonucleotides were hybridized.
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Vascular dysfunction in the offspring of AT1 receptor antibody-positive pregnant rats during high-salt diet.
Sheng Li Xue Bao
PUBLISHED: 04-21-2011
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Antibody against the angiotensin AT1 receptor (AT1-Ab) could disturb placental development. The placenta is the key organ between mother and fetus. Placental damage will seriously impair fetal growth and development in utero, leading to intrauterine growth restriction (IUGR). Based on the fetal origins of adult disease (FOAD) hypothesis, IUGR could increase a propensity to develop adult onset cardiovascular disease (CVD). The present study was designed to determine whether vascular function has changed in the adult offspring of AT1-Ab positive pregnant rats. Twenty four female rats (8-week-old, AT1-Ab negative) were randomly divided into two groups, immunized and vehicle groups. Immunized group received active immunization to establish AT1-Ab-positive model, while vehicle group was subjected to Freunds adjuvant without antigen. After 8 weeks of immunization, the antibody titers in sera from the female rats were detected by enzyme-linked immunosorbent assay (ELISA). Then all the female rats were mated with normal Wistar male rats and became pregnant. Immunized/vehicle group offspring rats (I offspring/V offspring) were raised to 40-week-old under standard chow feeding. Then the two groups offspring rats were given a high-salt diet for 12 weeks (4% NaCl in chow feeding). Systolic blood pressure (SBP) was measured dynamically by noninvasive blood pressure system. The vascular ring experiment was performed to detect vascular function and reactivity. As detected by ELISA, the titers of antibody peaked at the 8th week (OD values: 2.75 ± 0.08 vs 0.33 ± 0.01, P < 0.01 vs vehicle group at the same time point). There was no significant difference of SBP between the two groups offspring rats during the high-salt diet (P > 0.05). Isolated thoracic aortic rings of I offspring had significantly decreased constriction under norepinephrine treatment (P < 0.01 vs V offspring) and significantly decreased dilation under acetylcholine treatment (P < 0.05 vs V offspring). These results suggest that the offspring of AT1-Ab-positive pregnant rats are more susceptible to vascular functional abnormality while being fed high-salt diet.
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In vitro effects of polyethylene glycol in University of Wisconsin preservation solution on human red blood cell aggregation and hemorheology.
Clin. Hemorheol. Microcirc.
PUBLISHED: 04-19-2011
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Addition of hydroxyethyl starch (HES) to UW (University of Wisconsin) solution increases viscosity of the solution and red blood cell (RBC) aggregation. Recently, it was suggested that HES could be replaced by a new colloid, polyethylene glycol (PEG), in UW solution. The aim of this study was to see whether and how PEG affected RBC aggregation, and whether RBC aggregation parameters had any correlation with the molecular weight and concentration of PEG. After giving informed consent and signing consent documents, 12 healthy volunteers were enrolled in the study. Blood samples obtained from these volunteers were mixed with the test solutions with blood/solutions ratios of 5:1 and 1:1. Human RBC aggregation was investigated with an automatic hemorheological analyzer. Blood viscosity was measured with a cone-plate viscometer. Morphological characters of RBC aggregates were evaluated by light microscopy. It was found that viscosity was not affected by the Colloid-free UW solution. PEG20kDa (1 and 10 g/L) and PEG35kDa (1 g/L) had little effect on RBC aggregation, while PEG20kDa (30 g/L) and PEG35kDa (10 and 30 g/L) had a significant hyperaggregating effect on RBC. In conclusion, PEGs had a potential hyperaggregating effect on human RBC. This effect is correlated with PEG molecular weight and concentration. The use of large molecular weight and high concentration PEG in UW solution accounts for extended and accelerated aggregation of erythrocytes. The use of low concentration PEG35kDa (1 g/L) would be the optimal choice.
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Cross-linked DNA generated by "bis-click" reactions with bis-functional azides: site independent ligation of oligonucleotides via nucleobase alkynyl chains.
J. Org. Chem.
PUBLISHED: 11-11-2010
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Template-free cross-linking of single-stranded DNA bearing octadiynyl side chains at the 7-position of 8-aza-7-deazapurine moieties with bisfunctional azides is reported employing a Cu(I)-catalyzed azide-alkyne "bis-click" reaction. Bis-adducts were formed when the bis-azide:oligonucleotide ratio was 1:1; monofunctionalization occurred when the ratio was 15:1. Four-stranded DNA consisting of two cross-linked duplexes was obtained after hydridization. Cross-linked duplexes are as stable as individual duplexes when ligation was introduced at terminal positions; ligation at a central position led to a slight duplex destabilization.
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Spatially controlled DNA nanopatterns by "click" chemistry using oligonucleotides with different anchoring sites.
J. Am. Chem. Soc.
PUBLISHED: 10-13-2010
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DNA patterning on surfaces has broad applications in biotechnology, nanotechnology, and other fields of life science. The common patterns make use of the highly selective base pairing which might not be stable enough for further manipulations. Furthermore, the fabrication of well-defined DNA nanostructures on solid surfaces usually lacks chemical linkages to the surface. Here we report a template-free strategy based on "click" chemistry to fabricate spatially controlled DNA nanopatterns immobilized on surfaces. The self-assembly process utilizes DNA with different anchoring sites. The position of anchoring is of crucial importance for the self-assembly process of DNA and greatly influences the assembly of particular DNA nanopatterns. It is shown that the anchoring site in a central position generates tunable nanonetworks with high regularity, compared to DNAs containing anchoring sites at terminal and other positions. The prepared patterns may find applications in DNA capturing and formation of pores and channels and can serve as templates for the patterning using other molecules.
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Greatly enhanced energy density and patterned films induced by photo cross-linking of poly(vinylidene fluoride-chlorotrifluoroethylene).
Macromol Rapid Commun
PUBLISHED: 07-30-2010
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Greatly enhanced energy density in poly(vinylidene fluoride-chlorotrifluoroethylene) [P(VDF-CTFE)] is realized through interface effects induced by a photo cross-linking method. Being different from nanocomposites with lowered dielectric strength, the cross-linked P(VDF-CTFE)s possess a high breakdown field as well as remarkably elevated polarization, both of which contribute to the enhanced energy density as high as 22.5 J?·?cm(-3). Moreover, patterned thin films with various shapes and sizes are fabricated by photolithography, which sheds new light on the integration of PVDF-based electroactive polymers into organic microelectronic devices such as flexible pyroelectric/piezoelectric sensor arrays or non-volatile ferroelectric memory devices.
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[Study on the transmission of Hantaan virus and Orientia tsutsugamushi by naturally dual infected Leptotrombidium scutellare through stinging].
Zhonghua Yu Fang Yi Xue Za Zhi
PUBLISHED: 07-27-2010
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To investigate whether Leptotrombidium scutellare could be naturally infected by both Hantaan virus (HV) and Orientia tsutsugamushi (OT) and transmission status by stinging.
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The effect of emodin, an anthraquinone derivative extracted from the roots of Rheum tanguticum, against herpes simplex virus in vitro and in vivo.
J Ethnopharmacol
PUBLISHED: 07-22-2010
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Herpes simplex viruses (HSV-1 and -2) are important pathogens for humans and the discovery of novel anti-HSV drugs with low toxicity deserves great efforts. Rhubarb is one of the oldest and best-known traditional Chinese medicines. We initiated this study to test if emodin is the active ingredients from Rheum tanguticum (R. tanguticum, one of the Chinese Rhubarb) against HSV infection and to investigate its antiviral activity on HSV infection in tissue culture cells and in a mouse model.
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A novel mixed-mode solid phase extraction for simultaneous determination of melamine and cyanuric acid in food by hydrophilic interaction chromatography coupled to tandem mass chromatography.
J. Chromatogr. B Analyt. Technol. Biomed. Life Sci.
PUBLISHED: 07-11-2010
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Utilizing a solid phase extraction column (MCT) containing mixed hydrophilic functional gel and cation exchange sorbent, a sensitive and rapid HPLC-MS/MS method for simultaneously determining the residues of melamine (MEL) and cyanuric acid (CYA) in human foodstuffs was developed. MEL and CYA in egg, pork, liver, kidney and pork, shrimp, sausage casing, honey, soybean milk, soybean powder and dairy product were extracted using acetonitrile/water, defatted with hexane and isolated using MCT solid phase extraction column. The residues were separated upon a hydrophilic interaction liquid chromatography (HILIC) column and analyzed by electrospray ionization under negative-positive switched mode on a triplequadrupole mass spectrometry. The selected reaction monitoring was performed on [M+H](+) of m/z 127.9 to provide the transition of 127>85 and 127>68 (MEL) while the [M-H](-) of m/z 127.1 was selected as the precursor ion for CYA resulting in product ions m/z 85 and 42. Isotope labeled internal standard ((15)N(3)-MEL and (13)C(3)-CYA) and matrix-matched calibration were both used to observe the recovery to be 70.0-129.6% and 70.0-128.9% with RSD of 1.4-23.3% and 1.5-21.7% for MEL and CYA, respectively (n=6). All the LODs and LOQs of MEL and CYA were less than 39.4 and 99.1?gkg(-1), respectively, in 18 matrices, which were sensitive enough for quantitative analysis. This method has been proven effective in simultaneous determination of melamine and cyanuric acid when inspecting unknown and positive samples.
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Slit-Robo signaling mediates lymphangiogenesis and promotes tumor lymphatic metastasis.
Biochem. Biophys. Res. Commun.
PUBLISHED: 04-26-2010
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The Slit family of guidance cues binds to Roundabout (Robo) receptors to modulate neuronal, leukocytic, and endothelial migration. Slit-Robo signaling had been reported to function as chemoattractive signal for vascular endothelial cells during angiogenesis. In this study, we found that Robo1 was expressed in lymphatic endothelial cells to mediate the migration and tube formation of these cells upon Slit2 stimulation, which were specifically inhibited by the function-blocking antibody R5 to Slit2/Robo1 interaction. To further explore the lymphangiogenic effect and significance mediated by Slit-Robo signaling, we intercrossed Slit2 transgenic mice with a non-metastatic RIP1-Tag2 mouse tumor model, and found that transgenic overexpression of Slit2 significantly enhanced tumor lymphangiogenesis and subsequently promoted mesenteric lymph node metastasis of pancreatic islet tumors. Taken together, our findings reveal that through interacting with Robo1, Slit2 is a novel and potent lymphangiogenic factor and contributes to tumor lymphatic metastasis.
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NGAL and NGALR are frequently overexpressed in human gliomas and are associated with clinical prognosis.
J. Neurooncol.
PUBLISHED: 03-21-2010
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Recently, neutrophil gelatinase-associated lipocalin (NGAL) and its cell surface receptor, NGALR, have been shown to have critical roles in the biology of various tumors. Therefore, we investigated the expression of NGAL and NGALR in tumor sections obtained from patients with gliomas, and compared these results with the clinical characteristics of the patients. Using immunohistochemical assays, the expression levels of NGAL and NGALR were found to be up-regulated in tumor tissues, and to be related to tumor grade (p < 0.001). A positive correlation between expression of the two markers was also observed in these assays (r = 0.849; p < 0.001). Overexpression of NGAL and NGALR in glioma tissues was also confirmed in western blot analysis and real-time quantitative RT-PCR assays. Furthermore, overexpression of NGAL and NGALR was found to be significantly associated with poor prognosis (p < 0.001 in each case). Multivariate analysis identified patient age, tumor grade, and expression levels of NGAL and NGALR to be independent prognostic factors. In particular, NGAL(2+)/NGALR(2+) tissues were associated with lower rates of survival (risk ratio, 1.378; 95% CI, 1.102-1.724; p = 0.005). These findings suggest that NGAL and NGALR expression are frequently up-regulated in gliomas, and are closely associated with poor clinical outcome.
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The epidemiology and clinical management of craniocerebral injury caused by the Sichuan earthquake.
Neurol India
PUBLISHED: 03-16-2010
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Earthquake is one of the most devastating natural disasters that threaten human lives. Worldwide more than 3 million deaths have been caused by earthquakes in recent 20 years.
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Pathway-based genome-wide association analysis identified the importance of regulation-of-autophagy pathway for ultradistal radius BMD.
J. Bone Miner. Res.
PUBLISHED: 03-05-2010
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Wrist fracture is not only one of the most common osteoporotic fractures but also a predictor of future fractures at other sites. Wrist bone mineral density (BMD) is an important determinant of wrist fracture risk, with high heritability. Specific genes underlying wrist BMD variation are largely unknown. Most published genome-wide association studies (GWASs) have focused only on a few top-ranking single-nucleotide polymorphisms (SNPs)/genes and considered each of the identified SNPs/genes independently. To identify biologic pathways important to wrist BMD variation, we used a novel pathway-based analysis approach in our GWAS of wrist ultradistal radius (UD) BMD, examining approximately 500,000 SNPs genome-wide from 984 unrelated whites. A total of 963 biologic pathways/gene sets were analyzed. We identified the regulation-of-autophagy (ROA) pathway that achieved the most significant result (p = .005, q(fdr) = 0.043, p(fwer) = 0.016) for association with UD BMD. The ROA pathway also showed significant association with arm BMD in the Framingham Heart Study sample containing 2187 subjects, which further confirmed our findings in the discovery cohort. Earlier studies indicated that during endochondral ossification, autophagy occurs prior to apoptosis of hypertrophic chondrocytes, and it also has been shown that some genes in the ROA pathway (e.g., INFG) may play important roles in osteoblastogenesis or osteoclastogenesis. Our study supports the potential role of the ROA pathway in human wrist BMD variation and osteoporosis. Further functional evaluation of this pathway to determine the mechanism by which it regulates wrist BMD should be pursued to provide new insights into the pathogenesis of wrist osteoporosis.
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Genome-wide association study for femoral neck bone geometry.
J. Bone Miner. Res.
PUBLISHED: 02-23-2010
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Poor femoral neck bone geometry at the femur is an important risk factor for hip fracture. We conducted a genome-wide association study (GWAS) of femoral neck bone geometry, examining approximately 379,000 eligible single-nucleotide polymorphisms (SNPs) in 1000 Caucasians. A common genetic variant, rs7430431 in the receptor transporting protein 3 (RTP3) gene, was identified in strong association with the buckling ratio (BR, P = 1.6 x 10(-7)), an index of bone structural instability, and with femoral cortical thickness (CT, P = 1.9 x 10(-6)). The RTP3 gene is located in 3p21.31, a region that we found to be linked with CT (LOD = 2.19, P = 6.0 x 10(-4)) in 3998 individuals from 434 pedigrees. The replication analyses in 1488 independent Caucasians and 2118 Chinese confirmed the association of rs7430431 to BR and CT (combined P = 7.0 x 10(-3) for BR and P = 1.4 x 10(-2) for CT). In addition, 350 hip fracture patients and 350 healthy control individuals were genotyped to assess the association of the RTP3 gene with the risk of hip fracture. Significant association between a nearby common SNP, rs10514713 of the RTP3 gene, and hip fracture (P = 1.0 x 10(-3)) was found. Our observations suggest that RTP3 may be a novel candidate gene for femoral neck bone geometry.
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The protective effects of alpha-ketoacids against oxidative stress on rat spermatozoa in vitro.
Asian J. Androl.
PUBLISHED: 12-14-2009
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The aim of this study was to determine the effects of antioxidants, including alpha-ketoacids (alpha-ketoglutarate and pyruvate), lactate and glutamate/malate combination, against oxidative stress on rat spermatozoa. Our results showed that H(2)O(2) (250 micromol L(-1))-induced damages, such as impaired motility, adenosine triphosphate (ATP) depletion, inhibition of sperm protein phosphorylation, reduced acrosome reaction and decreased viability, could be significantly prevented by incubation of the spermatozoa with alpha-ketoglutarate (4 mmol L(-1)) or pyruvate (4 mmol L(-1)). Without exogenous H(2)O(2) in the medium, the addition of pyruvate (4 mmol L(-1)) significantly increased the superoxide anion (O(2)(-).) level in sperm suspension (P < or = 0.01), whereas the addition of alpha-ketoglutarate (4 mmol L(-1)) and lactate (4 mmol L(-1)) significantly enhanced tyrosine-phosphorylated proteins with the size of 95 kDa (P < or = 0.04). At the same time, alpha-ketoglutarate, pyruvate, lactate, glutamate and malate supplemented in media can be used as important energy sources and supply ATP for sperm motility. In conclusion, the present results show that alpha-ketoacids could be effective antioxidants for protecting rat spermatozoa from H(2)O(2) attack and could be effective components to improve the antioxidant capacity of Biggers, Whitten and Whittingham media.
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[Analysis of essential oil in herbal pair Artemisia annua-Agastache rugosa by GC-MS and chemometric resolution method].
Yao Xue Xue Bao
PUBLISHED: 11-01-2009
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Gas chromatograph-mass spectrometry, chemometric resolution method (CRM) and overall volume integration method were used to analyze the essential components of herbal pair Artemisia annua-Agastache rugosa (AA-AR) and compare it with that of single herbs AA and AR. The results showed that the components of volatile oil of herbal pair (AA-AR) were different from that of single herb drug in quality and quantity. 70, 69, and 48 essential components in essential oil of herbal pair (AA-AR), AA and AR were determined, accounting for about 85.93%, 88.85% and 93.23% of the total volatile oil, respectively. The volatile active components of the essential oils compounds in number are almost the sum of that of two single herbs, are mainly from herb AA, and the contents of each component from herb AR were relatively high. There are 51 common active constituents shared by herbal pair AA-AR and AA, and 34 common active constituents shared by herbal pair AA-AR and AR. There are 7 new components in the essential oils of herbal pair AA-AR, the relative content of arteannuic acid (2.99%) and p-propenyl-anisole (1.92%) are higher than others.
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Genome-wide association study identifies two novel loci containing FLNB and SBF2 genes underlying stature variation.
Hum. Mol. Genet.
PUBLISHED: 07-15-2009
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Human stature, as an important physical index in clinical practice and a usual covariate in gene mapping of complex disorders, is a highly heritable complex trait. To identify specific genes underlying stature, a genome-wide association study was performed in 1000 unrelated homogeneous Caucasian subjects using Affymetrix 500K arrays. A group of seven contiguous markers in the region of SBF2 gene (Set-binding factor 2) are associated with stature, significantly so at the genome-wide level after false discovery rate (FDR) correction (FDR q = 0.034-0.042). Three SNPs in another SNP group in the Filamin B (FLNB) gene were also associated with stature, significantly so with FDR q = 0.042-0.048. In follow-up independent replication studies, rs10734652 in the SBF2 gene was significantly (P = 0.036) and suggestively (P = 0.07) associated with stature in Caucasian families and 1306 unrelated Caucasian subjects, respectively, and rs9834312 in the FLNB gene was also associated with stature in such two independent Caucasian populations (P = 0.008 in unrelated sample and P = 0.049 in family sample). Particularly, additional significant replication association signals were detected in Chinese, an ethnic population different from Caucasian, between rs9834312 and stature in 619 unrelated northern Chinese subjects (P = 0.017), as well as between rs10734652 and stature in 2953 unrelated southern Chinese subjects (P = 0.048). This study also provides additional replication evidence for some of the already published stature loci. These results, together with the known functional relevance of the SBF2 and FLNB genes to skeletal linear growth and bone formation, support that two regions containing FLNB and SBF2 genes are two novel loci underlying stature variation.
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[Effect on bioactive components of Xuesaitong injection in the process of ultra-filter].
Zhong Yao Cai
PUBLISHED: 07-02-2009
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To study the effect on bioactive components of Xuesaitong injection in the process of ultra-filter by orthogonal test.
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Autophagic cell death induced by 5-FU in Bax or PUMA deficient human colon cancer cell.
Cancer Lett.
PUBLISHED: 04-19-2009
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Autophagy is a membrane process that results in the transporting of cellular contents to lysosomes for degradation. Autophagic cell death is another way of programed cell death called type II PCD, which has complicated connection with apoptosis, both of these two types of cell death play an important role in tumor development. In this study, we investigated chemotherapeutic agent induced cell death pathway in wild type (WT), Bax(-/-) and PUMA(-/-) HCT116 cells. Bax or PUMA deficient cells had similar chemosensitivity to WT cells but were defective in undergoing apoptosis. The results of electron microscopy and GFP-LC3 localization assay showed that autophagy was induced in Bax or PUMA deficient cells but not in WT cells. mTOR activity was decreased in Bax or PUMA deficient cells which further indicated the up-regulation of autophagy. Inhibition of autophagy by 3-Methyladenine (3-MA) decreased the cell death in Bax or PUMA deficient cells. Taken together, these results suggest that autophagic cell death can be used as an alternative cell death pathway in apoptosis defective cells and may bring a new target for cancer therapy.
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Pathway-based genome-wide association analysis identified the importance of EphrinA-EphR pathway for femoral neck bone geometry.
Bone
PUBLISHED: 04-14-2009
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Femoral neck (FN) bone geometry is an important predictor of bone strength with high heritability. Previous studies have revealed certain candidate genes for FN bone geometry. However, the majority of the underlying genetic factors remain to be discovered. In this study, pathway-based genome-wide association analysis was performed to explore the joint effects of genes within biological pathways on FN bone geometry variations in a cohort of 1000 unrelated US whites. Nominal significant associations (nominal p value<0.05) were observed between 76 pathways and a key FN bone geometry variable-section modulus (Z), biomechanically indicative of bone strength subject to bending. Among them, EphrinA-EphR pathway was most significantly associated with FN Z even after multiple testing adjustments (p(FWER) value=0.035). The association of EphrinA-EphR pathway with FN Z was also observed in an independent sample from Framingham Osteoporosis Study. Overall, these results suggest the significant genetic contribution of EphrinA-EphR pathway to femoral neck bone geometry.
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Association analyses of RANKL/RANK/OPG gene polymorphisms with femoral neck compression strength index variation in Caucasians.
Calcif. Tissue Int.
PUBLISHED: 04-09-2009
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Femoral neck compression strength index (fCSI), a novel phenotypic parameter that integrates bone density, bone size, and body size, has significant potential to improve hip fracture risk assessment. The genetic factors underlying variations in fCSI, however, remain largely unknown. Given the important roles of the receptor activator of the nuclear factor-kappaB ligand/receptor activator of the nuclear factor-kappaB/osteoprotegerin (RANKL/RANK/OPG) pathway in the regulation of bone remodeling, we tested the associations between RANKL/RANK/OPG polymorphisms and variations in fCSI as well as its components (femoral neck bone mineral density [fBMD], femoral neck width [FNW], and weight). This was accomplished with a sample comprising 1873 subjects from 405 Caucasian nuclear families. Of the 37 total SNPs studied in these three genes, 3 SNPs, namely, rs12585014, rs7988338, and rs2148073, of RANKL were significantly associated with fCSI (P = 0.0007, 0.0007, and 0.0005, respectively) after conservative Bonferroni correction. Moreover, the three SNPs were approximately in complete linkage disequilibrium. Haplotype-based association tests corroborated the single-SNP results since haplotype 1 of block 1 of the RANKL gene achieved an even more significant association with fCSI (P = 0.0003) than any of the individual SNPs. However, we did not detect any significant associations of these genes with fBMD, FNW, or weight. In summary, our findings suggest that the RANKL gene may play an important role in variation in fCSI, independent of fBMD and non-fBMD components.
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8-Aza-7-deazaguanine nucleosides and oligonucleotides with octadiynyl side chains: synthesis, functionalization by the azide-alkyne click reaction and nucleobase specific fluorescence quenching of coumarin dye conjugates.
Org. Biomol. Chem.
PUBLISHED: 03-02-2009
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Oligonucleotides incorporating 7-(octa-1,7-diynyl) derivatives of 8-aza-7-deaza-2-deoxyguanosine (2d) were prepared by solid-phase synthesis. The side chain of 2d was introduced by the Sonogashira cross coupling reaction and phosphoramidites (3a, 3b) were synthesized. Duplexes containing 2d are more stabilized compared to those incorporating the non-functionalized 8-aza-7-deaza-2-deoxyguanosine (2a) demonstrating that these side chains have steric freedom in duplex DNA. Nucleoside 2d as well as 2d-containing oligonucleotides were conjugated to the non-fluorescent 3-azido-7-hydroxycoumarin 15 by the Huisgen-Meldal-Sharpless click reaction. Pyrazolo[3,4-d]pyrimidine nucleoside conjugate 16 shows a much higher fluorescence intensity than that of the corresponding pyrrolo[2,3-d]pyrimidine derivative 17. The quenching in the dye conjugate 17 was found to be stronger on the stage of monomeric conjugates than in single-stranded or duplex DNA. Nucleobase-dye contact complexes are suggested which are more favourable in the monomeric state than in the DNA chain when the nucleobase is part of the stack. The side chains with the bulky dye conjugates are well accommodated in DNA duplexes thereby forming hybrids which are slightly more stable than canonical DNA.
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Genome-wide association analyses identify SPOCK as a key novel gene underlying age at menarche.
PLoS Genet.
PUBLISHED: 02-12-2009
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For females, menarche is a most significant physiological event. Age at menarche (AAM) is a trait with high genetic determination and is associated with major complex diseases in women. However, specific genes for AAM variation are largely unknown. To identify genetic factors underlying AAM variation, a genome-wide association study (GWAS) examining about 380,000 SNPs was conducted in 477 Caucasian women. A follow-up replication study was performed to validate our major GWAS findings using two independent Caucasian cohorts with 854 siblings and 762 unrelated subjects, respectively, and one Chinese cohort of 1,387 unrelated subjects--all females. Our GWAS identified a novel gene, SPOCK (Sparc/Osteonectin, CWCV, and Kazal-like domains proteoglycan), which had seven SNPs associated with AAM with genome-wide false discovery rate (FDR) q<0.05. Six most significant SNPs of the gene were selected for validation in three independent replication cohorts. All of the six SNPs were replicated in at least one cohort. In particular, SNPs rs13357391 and rs1859345 were replicated both within and across different ethnic groups in all three cohorts, with p values of 5.09 x 10(-3) and 4.37 x 10(-3), respectively, in the Chinese cohort and combined p values (obtained by Fishers method) of 5.19 x 10(-5) and 1.02 x 10(-4), respectively, in all three replication cohorts. Interestingly, SPOCK can inhibit activation of MMP-2 (matrix metalloproteinase-2), a key factor promoting endometrial menstrual breakdown and onset of menstrual bleeding. Our findings, together with the functional relevance, strongly supported that the SPOCK gene underlies variation of AAM.
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Genome-wide association and replication studies identified TRHR as an important gene for lean body mass.
Am. J. Hum. Genet.
PUBLISHED: 02-06-2009
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Low lean body mass (LBM) is related to a series of health problems, such as osteoporotic fracture and sarcopenia. Here we report a genome-wide association (GWA) study on LBM variation, by using Affymetrix 500K single-nucleotide polymorphism (SNP) arrays. In the GWA scan, we tested 379,319 eligible SNPs in 1,000 unrelated US whites and found that two SNPs, rs16892496 (p = 7.55 x 10(-8)) and rs7832552 (p = 7.58 x 10(-8)), within the thyrotropin-releasing hormone receptor (TRHR) gene were significantly associated with LBM. Subjects carrying unfavorable genotypes at rs16892496 and rs7832552 had, on average, 2.70 and 2.55 kg lower LBM, respectively, compared to those with alternative genotypes. We replicated the significant associations in three independent samples: (1) 1488 unrelated US whites, (2) 2955 Chinese unrelated subjects, and (3) 593 nuclear families comprising 1972 US whites. Meta-analyses of the GWA scan and the replication studies yielded p values of 5.53 x 10(-9) for rs16892496 and 3.88 x 10(-10) for rs7832552. In addition, we found significant interactions between rs16892496 and polymorphisms of several other genes involved in the hypothalamic-pituitary-thyroid and the growth hormone-insulin-like growth factor-I axes. Results of this study, together with the functional relevance of TRHR in muscle metabolism, support the TRHR gene as an important gene for LBM variation.
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Seoul virus and hantavirus disease, Shenyang, Peoples Republic of China.
Emerging Infect. Dis.
PUBLISHED: 02-06-2009
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An outbreak of hemorrhagic fever with renal syndrome (HFRS) occurred among students in Shenyang Pharmaceutical University in 2006. We conducted a study to characterize etiologic agents of the outbreaks and clarify the origin of hantaviruses causing infections in humans and laboratory animals. Immunoglobulin (Ig) M or IgG antibodies against Seoul virus (SEOV) were detected in the serum samples of all 8 patients. IgG antibodies against hantavirus were also identified in laboratory rats, which were used by these students for their scientific research. Phylogenetic analysis showed that partial small segment sequences recovered from humans, laboratory rats, and local wild rats belonged to SEOV. Hantavirus sequences recovered from humans and laboratory rats clustered within 1 of 3 lineages of SEOV circulating among local wild rats in Shenyang. These results suggest that the HFRS outbreak in Shenyang was caused by SEOV that was circulating among local wild rats and had also infected the laboratory rats.
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Genome-wide association and follow-up replication studies identified ADAMTS18 and TGFBR3 as bone mass candidate genes in different ethnic groups.
Am. J. Hum. Genet.
PUBLISHED: 01-17-2009
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To identify and validate genes associated with bone mineral density (BMD), which is a prominent osteoporosis risk factor, we tested 379,319 SNPs in 1000 unrelated white U.S. subjects for associations with BMD. For replication, we genotyped the most significant SNPs in 593 white U.S. families (1972 subjects), a Chinese hip fracture (HF) sample (350 cases, 350 controls), a Chinese BMD sample (2955 subjects), and a Tobago cohort of African ancestry (908 males). Publicly available Framingham genome-wide association study (GWAS) data (2953 whites) were also used for in silico replication. The GWAS detected two BMD candidate genes, ADAMTS18 (ADAM metallopeptidase with thrombospondin type 1 motif, 18) and TGFBR3 (transforming growth factor, beta receptor III). Replication studies verified the significant findings by GWAS. We also detected significant associations with hip fracture for ADAMTS18 SNPs in the Chinese HF sample. Meta-analyses supported the significant associations of ADAMTS18 and TGFBR3 with BMD (p values: 2.56 x 10(-5) to 2.13 x 10(-8); total sample size: n = 5925 to 9828). Electrophoretic mobility shift assay suggested that the minor allele of one significant ADAMTS18 SNP might promote binding of the TEL2 factor, which may repress ADAMTS18 expression. The data from NCBI GEO expression profiles also showed that ADAMTS18 and TGFBR3 genes were differentially expressed in subjects with normal skeletal fracture versus subjects with nonunion skeletal fracture. Overall, the evidence supports that ADAMTS18 and TGFBR3 might underlie BMD determination in the major human ethnic groups.
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[Meta-analysis on impact of Danshen on liver regeneration in rats].
Zhongguo Zhong Yao Za Zhi
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To assess the effect of Danshen on liver regeneration capacity of carbon tetrachloride-induced liver injury rats.
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Evidence for cell apoptosis suppressing white spot syndrome virus replication in Procambarus clarkii at high temperature.
Dis. Aquat. Org.
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In shrimp, higher water temperatures (~32°C) can suppress the ability of white spot syndrome virus (WSSV) to replicate and cause mortality, but the mechanisms remain unclear. To investigate whether cell apoptosis might be involved, a Tdt-mediated dUTP nick-end label (TUNEL) method was used to assess levels of chromosomal DNA fragmentation in hepatopancreas and gill cells of Procambarus clarkii crayfish infected with WSSV and maintained at either 32 ± 1°C or 24 ± 1°C. Based on relative cell numbers with yellow-green colored TUNEL-positive nuclei, the apoptotic index was elevated in WSSV-infected crayfish maintained at 32°C. In gill tissue sections examined by transmission electron microscope, cells with nuclei displaying apoptotic bodies or marginated, condensed and fragmented chromatin without concurrent cell cytoplasm damage were also more prevalent. Flow cytometry sorting of annexin-stained cells showed apoptosis to be most prevalent in granular haemocytes, and assays for caspase-3 activity showed it to be most elevated in hepatopancreas tissue. Despite these indicators of cell apoptosis but consistent with WSSV replication being restricted at elevated temperatures, no increases in transcription of the viral anti-apoptosis genes ORF390 and ORF222 were detected by RT-PCR in shrimp maintained at 32°C, possibly due to the elevated levels of cellular apoptosis.
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Establishment of SYBR green-based qPCR assay for rapid evaluation and quantification for anti-Hantaan virus compounds in vitro and in suckling mice.
Virus Genes
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Hantaan viruses cause two severe diseases lacking efficient treatment, yet no effective prophylactic vaccines are available. Continued exploration of alternative antiviral agents to treat hantavirus-related syndromes remains compulsory. The fluorescence-based quantitative real-time PCR (qPCR) has become the touchstone for target gene quantification. In the present study, standard curves for Hantaan virus (HTNV), mouse, and human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were generated by serial 10-fold dilutions of the constructed recombinant plasmid pGEM-T/HTNV, pGEM-T/mouse-GAPDH, and pGEM-T/human-GAPDH, respectively. Comparisons between the indirect immunofluorescence assay and qPCR assay in the detection of HTNV-infected Vero E6 cells showed improved detection limit and sensitivity of latter method. To characterize the inhibitory effect of several conventional antivirals (arbidol and ribavirin) and unconventional antivirals (indomethacin and curcumin) on HTNV, the levels of viral RNAs were measured for 4 days post-treatment of HTNV-infected Vero E6 cells and 18 days post-inoculation of HTNV-infected suckling mice. Our results validated that HTNV was sensitive to ribavirin and arbidol treatment, while indomethacin and curcumin may also be therapeutically effective in treating HTNV infection. As a result, the establishment and application of qPCR may be a useful tool for the evaluation of potential antivirals for Hantaan virus infection in vitro and in vivo.
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5-Ethynyl-2-deoxycytidine: a DNA building block with a clickable side chain.
Acta Crystallogr C
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The title compound [systematic name: 4-amino-1-(2-deoxy-?-D-erythro-pentofuranosyl)-5-ethynylpyrimidin-2(1H)-one], C(11)H(13)N(3)O(4), shows two conformations in the crystalline state. The N-glycosylic bonds of both conformers adopt similar conformations, with ? = -149.2 (1)° for conformer (I-1) and -151.4 (1)° for conformer (I-2), both in the anti range. The sugar residue of (I-1) shows a C2-endo envelope conformation ((2)E, S-type), with P = 164.7 (1)° and ?(m) = 36.9 (1)°, while (I-2) shows a major C3-exo sugar pucker (C3-exo-C2-endo, (3)T(2), S-type), with P = 189.2 (1)° and ?(m) = 33.3 (1)°. Both conformers participate in the formation of a layered three-dimensional crystal structure with a chain-like arrangement of the conformers. The ethynyl groups do not participate in hydrogen bonding, but are arranged in proximal positions.
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Amelioration of influenza virus-induced reactive oxygen species formation by epigallocatechin gallate derived from green tea.
Acta Pharmacol. Sin.
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To study whether epigallocatechin gallate (EGCG), a green tea-derived polyphenol, exerted anti-influenza A virus activity in vitro and in vivo.
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Caveolin-1 knockdown is associated with the metastasis and proliferation of human lung cancer cell line NCI-H460.
Biomed. Pharmacother.
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Caveolin-1 (CAV-1), one component of caveolae, involves in multiple cellular processes and signal transductions. We previously showed that the expression of CAV-1 gene in NCI-H446 cells inhibited cell proliferation and promoted cell metastasis. Here we explore the function of CAV-1 on tumor growth and metastasis by using NCI-H460 in vitro. First, we established NCI-H460 cell line, which CAV-1 was stably knockdown. Then we investigated the effects of CAV-1 on the morphology, proliferation, cell cycle and metastasis potential for NCI-H460 cell by crystal violet stains, CCK-8, colony formation, flow cytometry, scratch-wound assay and transwell assay. Western blot was used to examine the expression changes of cyclin D1, PCNA, E-cadherin and ?-catenin. Our results showed stable knockdown of CAV-1 inhibited the proliferation of NCI-H460 cells. Cell cycle of the transfected cells was arrested in G1/S phase and the expressions of cyclin D1 and PCNA protein were downregulated. Downregulation of CAV-1 promoted the migration and invasion abilities of NCI-H460 cells in vitro. The expression of ?-catenin increased and the level of E-cadherin decreased. In summary, our findings provide experimental evidence that CAV-1 may function as a proproliferative and antimetastatic gene in NCI-H460 cell line.
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The inhibitory effect of Rheum palmatum against coxsackievirus B3 in vitro and in vivo.
Am. J. Chin. Med.
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Coxsackievirus B(3)(CVB(3)) infection is the major cause of viral myocarditis, as well as dilated cardiomypathy. Rhubarb is one of the oldest and best-known traditional Chinese medicines. We initiated this study to determine the antiviral effect of an ethanol extract from the roots and rhizoma of Rheum palmatum (R. palmatum, one of the Chinese Rhubarbs), against CVB(3) in tissue culture cells and in a mouse model. The ethanol extract from R. palmatum showed significant inhibitory activity against CVB(3) on HEp-2 cells when added after infection, with IC(50) of 4 ?g/ml, TI of 10. The medicated mouse serum still contained the pharmaceutical compound 24 h after intraperitoneal injection, and exhibited an antiviral effect on CVB(3)-infected cells, especially in the 0.3 and 0.5 g/kg/day treatment groups. Furthermore, the CVB(3)-infected mice were treated with the extract solution with dosages of 0.3 g/kg/day beginning 24 h post-CVB(3) exposures. The ethanol extract treated mice showed alleviated clinical signs, better survival rate, prolonged MTD and decreased viral titers compared to the virus control group. Our results indicate that the ethanol extract from R. palmatum has the anti-CVB(3) activity in vitro and in vivo and thus provides a re-evaluation of this old remedy with a broad therapeutic potential.
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[Measuring diet quality of labor workers in Shenzhen using Chinese diet balance index].
Zhonghua Yu Fang Yi Xue Za Zhi
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To evaluate the overall diet quality and diet model of labor workers in Shenzhen using Chinese Diet Balance Index (DBI).
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Genetic characterization of a new subtype of Hantaan virus isolated from a hemorrhagic fever with renal syndrome (HFRS) epidemic area in Hubei Province, China.
Arch. Virol.
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To characterize hantaviruses currently circulating in the hemorrhagic fever with renal syndrome (HFRS) epidemic area of Hubei Province, rodents were captured and serum samples were collected from several HFRS patients. The partial S segment of the hantaviruses amplified from two serum samples had a high degree of sequence identity to the corresponding hantavirus strain isolated from Apodemus agrarius (designated as HV004). The complete S, M, and L segment sequences of HV004 were determined. The sequence identities between strain HV004 and other Hantaan viruses (HTNVs) were 83 %-90 % at the nucleotide level and 95 %-99 % at the amino acid level. Phylogenetic analysis showed that HV004 belonged to a new HTNV lineage. These data suggest the presence of a new HTNV subtype, which probably caused the HFRS cases in the endemic area of Hubei Province.
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ANKRD7 and CYTL1 are novel risk genes for alcohol drinking behavior.
Chin. Med. J.
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Alcohol dependence (AD) is a complex disorder characterized by impaired control over drinking. It is determined by both genetic and environmental factors. The recent approach of genome-wide association study (GWAS) is a powerful tool for identifying complex disease-associated susceptibility alleles, however, a few GWASs have been conducted for AD, and their results are largely inconsistent. The present study aimed to screen the loci associated with alcohol-related phenotypes using GWAS technology.
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Cross-Linked DNA: Site-Selective "Click" Ligation in Duplexes with Bis-Azides and Stability Changes Caused by Internal Cross-Links.
Bioconjug. Chem.
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Heterodimeric interstrand cross-linked DNA was constructed by the "bis-click" reaction carried out on preformed oligonucleotide duplexes with the bis-azide 1. For this, alkynylated 8-aza-7-deazapurine or corresponding 5-substituted pyrimidine nucleosides were synthesized. Cross-linking resulted in chemoselective formation of heterodimeric duplexes while homodimers were suppressed. For product identification, heterodimeric DNA was prepared by the "stepwise click" reaction, while noncomplementary homodimers were accessible by "bis-click" chemistry, unequivocally. Studies on duplex melting of complementary cross-linked duplexes (heterodimers) revealed significantly increased Tm values compared to the non-cross-linked congeners. The stability of this cross-linked DNA depends on the linker length and the site of modification. Cross-linked homodimers hybridized with single-stranded complementary oligonucleotides show much lower stability.
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Higher expression of Caveolin-1 inhibits human small cell lung cancer (SCLC) apoptosis in vitro.
Cancer Invest.
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Small cell lung cancer (SCLC) is the most aggressive type of lung cancer, and its treatment is closely associated with apoptosis. Caveolin-1 plays an important role in the development of a variety of human cancers. This study sought to investigate the influence of Caveolin-1 on the apoptosis of SCLC in vitro. We demonstrate that higher expression of Caveolin-1 leads to inhibition of cisplatin and Ultraviolet Radiation (UVR)-induced apoptosis in SCLC cells; and also could decrease caspase-3 activity and increase the stability of Bcl-2 at the protein level. Our findings illuminate a potential molecular mechanism regarding CAV-1s role as anti-apoptosis protein.
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A 2-deoxycytidine long-linker click adduct forming two conformers in the asymmetric unit.
Acta Crystallogr C
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The title compound {systematic name: 4-amino-1-(2-deoxy-?-D-erythro-pentofuranosyl)-5-[6-(1-benzyl-1H-1,2,3-triazol-4-yl)hex-1-ynyl]pyrimidin-2(1H)-one}, C(24)H(28)N(6)O(4), shows two conformations in the crystalline state, viz. (I-1) and (I-2). The pyrimidine groups and side chains of the two conformers are almost superimposable, while the greatest differences between them are observed for the sugar groups. The N-glycosylic bonds of both conformers adopt similar anti conformations, with ? = -168.02?(12)° for conformer (I-1) and ? = -159.08?(12)° for conformer (I-2). The sugar residue of (I-1) shows an N-type (C3-endo) conformation, with P = 33.1?(2)° and ?(m) = 29.5?(1)°, while the conformation of the 2-deoxyribofuranosyl group of (I-2) is S-type (C3-exo), with P = 204.5?(2)° and ?(m) = 33.8?(1)°. Both conformers participate in hydrogen-bond formation and exhibit identical patterns resulting in three-dimensional networks. Intermolecular hydrogen bonds are formed with neighbouring molecules of different and identical conformations (N-H...N, N-H... O, O-H...N and O-H...O).
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Construction and assembly of branched Y-shaped DNA: "click" chemistry performed on dendronized 8-aza-7-deazaguanine oligonucleotides.
Bioconjug. Chem.
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Branched DNA was synthesized from tripropargylated oligonucleotides by the Huisgen-Meldal-Sharpless cycloaddition using "stepwise and double click" chemistry. Dendronized oligonucleotides decorated with 7-tripropargylamine side chains carrying two terminal triple bonds were further functionalized with bis-azides to give derivatives with two terminal azido groups. Then, the branched side chains with two azido groups or two triple bonds were combined with DNA-fragments providing the corresponding clickable function. Both concepts afforded branched (Y-shaped) three-armed DNA. Annealing of branched DNA with complementary oligonucleotides yielded supramolecular assemblies. The concept of "stepwise and double click" chemistry combined with selective hybridization represents a flexible tool to generate DNA nanostructures useful for various purposes in DNA diagnostics, delivery, and material science applications.
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Screening and identification of distant metastasis-related differentially expressed genes in human squamous cell lung carcinoma.
Anat Rec (Hoboken)
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Distant metastasis is one of the leading causes of lung cancer death. Detecting the early-stage molecular alternations in primary tumors, such as gene expression differences, provides a "prognostic" value to the precaution of tumor metastasis. The aim of this article is to screen and identify the metastasis-related genes in human squamous cell lung carcinoma. Primary tumor tissues of nine patients with subsequent metastasis and eight patients without metastasis were selected to perform the gene microarray experiment. GO and pathway analyses were used to determine the differentially expressed genes. Two identified genes were further validated by real-time quantitative reverse transcription polymerase chain reaction (PCR) (real-time qRT-PCR). Two hundred and thirty-eight differentially expressed genes were detected in gene chip experiment, including 51 up-regulated genes and 187 down-regulated genes. These genes were involved in several cellular processes, including cell adhesion, cell cycle regulation, and apoptosis. GO analysis showed that the differentially expressed genes participated in a wide ranging of metastasis-related processes, including extracellular region and regulation of liquid surface tension. In addition, pathway analysis demonstrated that the differentially expressed genes were enriched in pathways related to cell cycle and Wnt signaling. Real-time qRT-PCR validation experiment of LCN2 and PDZK1IP1 showed a consistent up-regulation in the metastasis group. The metastasis of human squamous cell lung carcinoma is a complex process that is regulated by multiple gene alternations on the expression levels. The 238 differentially expressed genes identified in this study presumably contain a core set of genes involved in tumor metastasis. The real-time qRT-PCR results of PDZK1IP1 and LCN2 validated the reliability of this gene microarray experiment.
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