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Find video protocols related to scientific articles indexed in Pubmed.
The potential signaling pathway between peroxisome proliferator-activated receptor gamma and retinoic acid receptor alpha in renal interstitial fibrosis disease.
J. Recept. Signal Transduct. Res.
PUBLISHED: 11-01-2014
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Abstract Peroxisome proliferator-activated receptor? (PPAR?) can regulate the process of cell apoptosis and is related to the progression of renal disorders. Retinoic acid receptor alpha (RAR?) is one of the nuclear receptors involved in a variety of kidney diseases. Renal interstitial fibrosis (RIF) is a common denominator of chronic kidney disease (CKD). This study investigated whether a potential signaling pathway existed between PPAR? and RAR? in RIF rats with unilateral ureteral obstruction (UUO). The rats were randomly divided into four groups: a model group subjected to UUO (GU), and three other groups treated with rosiglitazone sodium (GRS), GW9662 and dimethyl sulfoxide (DMSO), n?=?40, respectively. Renal tissues were collected two and four weeks after post-surgery. The relevant indicators were detected. In comparison with the GU group, the expressions of PPAR? and RAR? (protein and mRNA) were increased in the GRS group, and decreased in the GW9662 group (all p?
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[Exploration and practice of research-based teaching in the course of theory of Meridians and Acupoints for acupuncture and tuina specialty].
Zhongguo Zhen Jiu
PUBLISHED: 10-23-2014
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For the purpose of exploring the teaching reform model and method, also promoting the quality of talents in acupuncture and tuina field, the research-based teaching model is applied into the course of Theory of Meridians and Acupoints. This method includes two parts of teaching and learning. For teachers, they bring modern research focus and trend into teaching through questionnaire survey among students, aiming to integrate the education inside and outside class. For students, they are guided to resolve the opening, enlightening and scalable issues through consulting abundant resources of literature and database to achieve autonomous participation in the course of learning. By this teaching method, it is expected to train the student's ability of expanding their thinking, as well as discovering, analyzing and solving the problem.
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Comparative expression profiling reveals gene functions in female meiosis and gametophyte development in Arabidopsis.
Plant J.
PUBLISHED: 09-30-2014
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Megasporogenesis is essential for female fertility, and requires the accomplishment of meiosis and the formation of functional megaspores. The inaccessibility and low abundance of female meiocytes make it particularly difficult to elucidate the molecular basis underlying megasporogenesis. We used high-throughput tag-sequencing analysis to identify genes expressed in female meiocytes (FMs) by comparing gene expression profiles from wild-type ovules undergoing megasporogenesis with those from the spl mutant ovules, which lack megasporogenesis. A total of 862 genes were identified as FMs, with levels that are consistently reduced in spl ovules in two biological replicates. Fluorescence-assisted cell sorting followed by RNA-seq analysis of DMC1:GFP-labeled female meiocytes confirmed that 90% of the FMs are indeed detected in the female meiocyte protoplast profiling. We performed reverse genetic analysis of 120 candidate genes and identified four FM genes with a function in female meiosis progression in Arabidopsis. We further revealed that KLU, a putative cytochrome P450 monooxygenase, is involved in chromosome pairing during female meiosis, most likely by affecting the normal expression pattern of DMC1 in ovules during female meiosis. Our studies provide valuable information for functional genomic analyses of plant germline development as well as insights into meiosis.
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[Hydrophilic interaction chromatography on silica column: retention mechanism and its influential factors].
Se Pu
PUBLISHED: 09-27-2014
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Hydrophilic interaction chromatography (HILIC) is a valuable alternative to reversed phase liquid chromatography (RPLC) for the analysis of highly polar and hydrophilic compounds, in which the separation mechanism is quite different from RPLC and the separation selectivity is complementary to RPLC. This separation mode can be characterized as normal phase liquid chromatography (NPLC) on polar columns in aqueous-organic mobile phases rich in organic solvents (usually acetonitrile). Silica has been the earliest developed and most widely used HILIC stationary phase. This review deals with the recent advances in the development of the retention mechanism on silica column with special attention to the effects of stationary phase, mobile phase composition and temperature on separation in HILIC mode. Moreover, the developing trends and applications of this HILIC mode are presented.
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Simultaneous adsorption of SO2 and NO from flue gas over mesoporous alumina.
Environ Technol
PUBLISHED: 09-05-2014
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Mesoporous alumina (MA) with a higher ability to simultaneously remove SO2 and NO was prepared by the evaporation-induced self-assembly process. The adsorption capacities of MA are 1.79 and 0.702?mmol/g for SO2 and NO, respectively. The Brunauer-Emmett-Teller method was used to characterize the adsorbent. Simultaneous adsorption of SO2 and NO from flue gas over MA in different operating conditions had been studied in a fixed bed reactor. The effects of temperature, oxygen concentration and water vapour were investigated. The experimental results showed that the optimum temperature for MA to simultaneously remove SO2 and NO was 90°C. The simultaneous adsorption capacities of SO2 and NO could be enhanced by increasing O2 when its concentration was below 5%. The changes of simultaneous adsorption capacities were not obvious when O2 concentration was above 5%. The increase in relative humidity results in an increase after dropping of SO2 adsorption capacity, whereas the adsorption capacity of NO showed an opposite trend. The results suggest that MA is a great adsorbent for simultaneous removal of SO2 and NO from flue gas.
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Dose of incorporated immunodominant antigen in recombinant BCG impacts modestly on Th1 immune response and protective efficiency against Mycobacterium tuberculosis in mice.
J Immunol Res
PUBLISHED: 07-23-2014
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One approach for improving BCG efficacy is to utilize BCG as vehicle to develop recombinant BCG (rBCG) strains overexpressing Mycobacterium tuberculosis (M. tb) antigens. Also expression level of a candidate antigen should impact the final T cell responses conferred by rBCG. In this study, based on our previously constructed differential expression system, we developed two rBCG strains overexpressing M. tb chimeric antigen Ag856A2 (coding a recombinant ag85a with 2 copies of esat-6 inserted at Acc I site of ag85a) at differential levels under the control of the subtly modified furA promoters. These two rBCG strains were used to vaccinate C57BL/6 mice and exploit dose of incorporated antigen in rBCG to optimize immune response and protective efficiency against M. tb challenge in mouse model. The results showed that rBCG strains overexpressing Ag856A2 at differential levels induced different antigen-specific IFN-? production and comparable number of M. tb-specific CD4 T cells expressing IL-2. M. tb challenge experiment showed that rBCG strains afforded enhanced but comparable immune protection characterized by reduced bacillary load, lung pathology, and inflammation. These results suggested that the dose of antigens incorporated in rBCG can impact T cell immune responses but imposed no significantly differential protective efficacies.
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Seroprevalence and risk factors of Toxoplasma gondii infection in domestic sika deer (Cervus nippon) in northeastern China.
Acta Trop.
PUBLISHED: 07-14-2014
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Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii, which can infect warm-blooded animals and humans. A serological survey was undertaken to examine the seroprevalence and risk factors associated with T. gondii infection in sika deer in northeastern China. 114 (13.46%, 95% CI 11.16-15.76) out of 847 serum samples were positive to T. gondii by modi?ed agglutination test (MAT) at a 1:25 cut-off, with titers of 1:25 in 44, 1:50 in 32, 1:100 in 17, 1:500 in 11, 1:1500 or higher in 10. These samples were collected between November 2012 and October 2013 from Inner Mongolia, Jilin and Heilongjiang provinces in China. However, statistically signi?cant differences were not observed between T. gondii seroprevalence and genders or regions of sika deer in the logistic regression analysis (P>0.05) and left out of the ?nal model. Seroprevalence of T. gondii infection in male sika deer was 14.07% (95% CI 11.14-17.01), slightly higher than that in the female (12.38%) (95% CI 8.69-16.06) and seroprevalence of T. gondii infection in Harbin, Changchun city, Jilin city and Chifeng city were 12.02% (95% CI 7.60-16.44), 15.51% (95% CI 11.52-19.50), 12.27% (95% CI 7.23-17.31) and 12.50% (95% CI 7.38-17.63), respectively. Seasons of sampling were considered as main risk factors associated with T. gondii infection, autumn (15.32%) were more than two times (OR=1.98, 95% CI=1.18-3.33, P=0.01) at risk of acquiring T. gondii infection compared to winter (8.37%). Our results indicated a widespread exposure to T. gondii among sika deer in China. To our knowledge, this is the first report of T. gondii seroprevalence in sika deer in China.
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Bone marrow stromal cells protect acute myeloid leukemia cells from anti-CD44 therapy partly through regulating PI3K/Akt-p27(Kip1) axis.
Mol. Carcinog.
PUBLISHED: 07-13-2014
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The anti-CD44 monoclonal antibody (mAb) A3D8 induces differentiation or apoptosis in vitro in various subtypes of acute myeloid leukemia (AML) via p27(Kip1) upregulation. Bone marrow (BM) stromal cells play a vital role in the development of chemoresistance in AML cells attached to the stroma. To investigate the effect of BM stroma adhesion induced AML resistance to A3D8, we developed a co-culture system composed of an AML-derived cell line (NB4) cultured with either a human BM stroma cell line (HS-5) or mesenchymal stem cells (MSCs). We found that NB4 cells adhered to HS-5 cells or MSCs developed resistance against the anti-proliferative effects of A3D8, and this action is caused by the activation of PI3K/Akt signaling following p27(Kip1) down-regulation and cytoplasmic re-localization. The stromal co-culture-induced resistance can be partially abolished by inhibiting the PI3K/Akt signaling pathway. Such findings were confirmed in two additional AML-derived cell lines as well as in primary AML cells. Our results suggest that BM stroma can induce A3D8 resistance in part via the PI3K/Akt-p27(Kip1) axis, and blocking PI3K/Akt pathway maybe necessary for anti-CD44 treatment on AML in BM microenvironment. © 2014 The Authors. Molecular Carcinogenesis published by Wiley Periodicals, Inc.
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Seroprevalence and Risk Factors of Chlamydia abortus Infection in Tibetan Sheep in Gansu Province, Northwest China.
ScientificWorldJournal
PUBLISHED: 06-27-2014
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Chlamydia abortus, an important pathogen in a variety of animals, is associated with abortion in sheep. In the present study, 1732 blood samples, collected from Tibetan sheep between June 2013 and April 2014, were examined by the indirect hemagglutination (IHA) test, aiming to evaluate the seroprevalence and risk factors of C. abortus infection in Tibetan sheep. 323 of 1732 (18.65%) samples were seropositive for C. abortus antibodies at the cut-off of 1?:?16. A multivariate logistic regression analysis was used to evaluate the risk factors associated with seroprevalence, which could provide foundation to prevent and control C. abortus infection in Tibetan sheep. Gender of Tibetan sheep was left out of the final model because it is not significant in the logistic regression analysis (P > 0.05). Region, season, and age were considered as major risk factors associated with C. abortus infection in Tibetan sheep. Our study revealed a widespread and high prevalence of C. abortus infection in Tibetan sheep in Gansu province, northwest China, with higher exposure risk in different seasons and ages and distinct geographical distribution.
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Palladium-catalyzed thiolation of alkanes and ethers with arylsulfonyl hydrazides.
Chem. Commun. (Camb.)
PUBLISHED: 06-24-2014
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A new method for the preparation of alkyl aryl sulfides through direct oxidative thiolation of alkanes or ethers with arylsulfonyl hydrazides using di-tert-butyl peroxide (DTBP) as an oxidant catalyzed by Pd(OAc)2 has been reported. The C-H bonds in various alkanes or ethers were successfully converted into C-S bonds to yield the corresponding sulfides in moderate to good yields.
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GSTT1 polymorphism and the risk of developing prostate cancer.
Am. J. Epidemiol.
PUBLISHED: 06-06-2014
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A possible association between glutathione S-transferase theta 1 gene (GSTT1) polymorphism and the risk of developing prostate cancer is currently hotly debated, but evidence from various epidemiologic studies remains unclear. This investigation was performed to assess whether an association between GSTT1 polymorphism and prostate cancer risk exists by using meta-analysis to combine comparable studies, thereby increasing sample size and statistical significance, as well as to identify patterns in various studies. The association reports were identified from the PubMed database and the Cochrane Library on March 1, 2013, and data from eligible studies (from 1999-2012) were synthesized. Thirty-eight reports were included in this meta-analysis on the association of the null genotype of GSTT1 with prostate cancer risk. No solid association between the GSTT1 null genotype and prostate cancer risk could be established for the overall population (odds ratio = 1.11, 95% confidence interval: 0.97, 1.27; P = 0.13). However, the GSTT1 null genotype was distinctly associated with prostate cancer risk in Caucasians (odds ratio = 1.24, 95% confidence interval: 1.03, 1.48, P = 0.02). In conclusion, the GSTT1 null genotype is associated with prostate cancer risk in Caucasians, but not in the overall population.
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A TIGAR-regulated metabolic pathway is critical for protection of brain ischemia.
J. Neurosci.
PUBLISHED: 05-30-2014
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TP53-induced glycolysis and apoptosis regulator (TIGAR) inhibits glycolysis and increases the flow of pentose phosphate pathway (PPP), which generates NADPH and pentose. We hypothesized that TIGAR plays a neuroprotective role in brain ischemia as neurons do not rely on glycolysis but are vulnerable to oxidative stress. We found that TIGAR was highly expressed in brain neurons and was rapidly upregulated in response to ischemia/reperfusion insult in a TP53-independent manner. Overexpression of TIGAR in normal mice with lentivirus reduced ischemic neuronal injury, whereas lentivirus-mediated TIGAR knockdown aggravated it. In cultured primary neurons, increasing TIGAR expression reduced oxygen and glucose deprivation (OGD)/reoxygenation-induced injury, whereas decreasing its expression worsened the injury. The glucose 6-phosphate dehydrogenase was upregulated in mouse and cellular models of stroke, and its upregulation was further enhanced by overexpression of TIGAR. Supplementation of NADPH also reduced ischemia/reperfusion brain injury and alleviated TIGAR knockdown-induced aggravation of ischemic injury. In animal and cellular stroke models, ischemia/reperfusion increased mitochondrial localization of TIGAR. OGD/reoxygenation-induced elevation of ROS, reduction of GSH, dysfunction of mitochondria, and activation of caspase-3 were rescued by overexpression of TIGAR or supplementation of NADPH, while knockdown of TIGAR aggravated these changes. Together, our results show that TIGAR protects ischemic brain injury via enhancing PPP flux and preserving mitochondria function, and thus may be a valuable therapeutic target for ischemic brain injury.
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Genome-wide identification and expression profiling of the SnRK2 gene family in Malus prunifolia.
Gene
PUBLISHED: 05-25-2014
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Sucrose non-fermenting-1-related protein kinase 2 (SnRK2) constitutes a small plant-specific serine/threonine kinase family with essential roles in the abscisic acid (ABA) signal pathway and in responses to osmotic stress. Although a genome-wide analysis of this family has been conducted in some species, little is known about SnRK2 genes in apple (Malus domestica). We identified 14 putative sequences encoding 12 deduced SnRK2 proteins within the apple genome. Gene chromosomal location and synteny analysis of the apple SnRK2 genes indicated that tandem and segmental duplications have likely contributed to the expansion and evolution of these genes. All 12 full-length coding sequences were confirmed by cloning from Malus prunifolia. The gene structure and motif compositions of the apple SnRK2 genes were analyzed. Phylogenetic analysis showed that MpSnRK2s could be classified into four groups. Profiling of these genes presented differential patterns of expression in various tissues. Under stress conditions, transcript levels for some family members were up-regulated in the leaves in response to drought, salinity, or ABA treatments. This suggested their possible roles in plant response to abiotic stress. Our findings provide essential information about SnRK2 genes in apple and will contribute to further functional dissection of this gene family.
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The role of retinoic acid receptors in the signal pathway of all-trans retinoic acid-induced differentiation in adriamycin-induced podocyte injury.
J. Recept. Signal Transduct. Res.
PUBLISHED: 05-20-2014
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Abstract All-trans retinoic acid (ATRA) plays an essential role in cell survival and differentiation by binding to retinoic acid receptors (RARs), including RAR-?, RAR-?, and RAR-?. Injury to podocytes is the most frequent cause of glomerulosclerosis (GS). This study was performed to investigate which of the RAR subtypes is involved in the signal pathway of ATRA-induced differentiation of injured podocytes. ATRA (0.1??M) was administered to Adriamycin (ADR)-induced, injured podocytes, in vitro. Morphological changes were observed. The protein/mRNA expression of podocin, nephrin, transforming growth factor ?1(TGF-?1), and the RARs (RAR-?,?,?) was measured by RT-PCR and Western blotting. ATRA treatment ameliorated cell hypertrophy and reduced the shedding of the cytoplasm which was observed under light microscope and the extension of the foot processes was observed under scan electron microscope. Compared with the injured podocytes, ATRA exposure significantly increased the protein/mRNA expression of nephrin and podocin and it markedly reduced TGF-?1 (all p??0.05). In conclusion, RAR-?/? (and RAR-? to a lesser degree) may be involved in the signal pathway of ATRA-induced differentiation in injured podocytes.
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Distribution of pathological finding in the children with nephrotic syndrome from Guangxi.
Saudi J Kidney Dis Transpl
PUBLISHED: 05-14-2014
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To identify the variations in pediatric renal biopsy pathology and clinicopathological features in Guangxi, China, in the past ten years, we studied retrospectively the kidney biopsies performed to evaluate the primary nephrotic syndrome (PNS) in 218 children at two main medical centers in Guangxi from January 1999 to January 2009. The major pathological finding was mesangial proliferative glomerulonephritis (48.2%), focal segmental glomerulosclerosis (16.5%), immunoglobulin A nephropathy (13.3%) and minimal change disease (11.0%). Patients with different pathological types yielded different response rates to glucocorticoids (P <0.001). There were statistical significant differences between prognosis for the different pathological types (P <0.05). The pathological characteristics of PNS in children were diverse and significant for guiding the grade of glucocorticoid response and predicting the prognosis of the PNS disease.
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LIM homeobox transcription factor 1B expression affects renal interstitial fibrosis and apoptosis in unilateral ureteral obstructed rats.
Am. J. Physiol. Renal Physiol.
PUBLISHED: 04-30-2014
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LIM homeobox transcription factor 1B (LMX1B) is a transcription factor of the LIM homeodomain type and has been implicated in the development of diverse structures such as limbs, kidneys, eyes, and the brain. Furthermore, LMX1B has been implicated in nail-patella syndrome, which is predominantly characterized by malformation of limbs and nails, and in 30% of patients, nephropathy, including renal fibrosis, is observed. Since no reports were available that studied the link between LMX1B expression and renal interstitial fibrosis, we explored if LMX1B affects typical markers of fibrosis, e.g., extracellular matrix components, profibrotic factors, and apoptosis as the final detrimental consequence. We recently showed that LMX1B acts as a negative regulator of transforming growth factor-?l, collagen type III, fibronectin, cleaved caspase-3, and the cell apoptosis rate in a renal tubular epithelial cell system under hypoxic conditions. Here, we confirmed these results in unilateral ureteral obstructed rats. Furthermore, LMX1B was distinctly expressed throughout the glomerulus and tubule lining, including epithelial cells. Knockdown of LMX1B aggravated the expression of fibrosis markers, oxidative stress, and apoptosis compared with the already increased levels due to unilateral ureteral obstruction, whereas overexpression attenuated these effects. In conclusion, reduced LMX1B levels clearly represent a risk factor for renal fibrosis, whereas overexpression affords some level of protection. In general, LMX1B may be considered to be a negative regulator of the fibrosis index, transforming growth factor-?l, collagen type III, fibronectin, cleaved caspase-3, cell apoptosis, ROS, and malondialdehyde (r = -0.756, -0.698, -0.921, -0.923, -0.843, -0.794, -0.883, and -0.825, all P < 0.01).
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ACTIN-RELATED PROTEIN6 Regulates Female Meiosis by Modulating Meiotic Gene Expression in Arabidopsis.
Plant Cell
PUBLISHED: 04-15-2014
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In flowering plants, meiocytes develop from subepidermal cells in anthers and ovules. The mechanisms that integrate gene-regulatory processes with meiotic programs during reproductive development remain poorly characterized. Here, we show that Arabidopsis thaliana plants deficient in ACTIN-RELATED PROTEIN6 (ARP6), a subunit of the SWR1 ATP-dependent chromatin-remodeling complex, exhibit defects in prophase I of female meiosis. We found that this meiotic defect is likely due to dysregulated expression of meiotic genes, particularly those involved in meiotic recombination, including DMC1 (DISRUPTED MEIOTIC cDNA1). Analysis of DMC1 expression in arp6 mutant plants indicated that ARP6 inhibits expression of DMC1 in the megasporocyte and surrounding nonsporogeneous ovule cells before meiosis. After cells enter meiosis, however, ARP6 activates DMC1 expression specifically in the megasporocyte even as it continues to inhibit DMC1 expression in the nonsporogenous ovule cells. We further show that deposition of the histone variant H2A.Z, mediated by the SWR1 chromatin-remodeling complex at the DMC1 gene body, requires ARP6. Therefore, ARP6 regulates female meiosis by determining the spatial and temporal patterns of gene expression required for proper meiosis during ovule development.
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Platelet-derived growth factor-BB induces matrix metalloproteinase-2 expression and rat vascular smooth muscle cell migration via ROCK and ERK/p38 MAPK pathways.
Mol. Cell. Biochem.
PUBLISHED: 04-12-2014
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Matrix metalloproteinases (MMP) play a pivotal role in the pathogenesis of cardiovascular diseases. Their expressions are altered in response to a variety of stimuli, including growth factors, inflammatory markers, and cytokines. In this study, we demonstrated that platelet-derived growth factor-BB (PDGF-BB) induces a dose- and time-dependent increase in MMP-2 expression in rat vascular smooth muscle cells (VSMC). Treatment with either the Rho-associated protein kinase (ROCK) inhibitor Y-27632 or suppression of ROCK-1/2 by small interfering RNA technology significantly reduced the MMP-2 expression, thus suggesting that ROCK regulates such expression. Similar results were observed when VSMC were pretreated with either U0126 or SB203580, which are selective inhibitors of extracellular signal-regulated kinase and p38 mitogen-activated protein kinase, respectively, thus suggesting that these kinases are important for the induction of MMP-2 expression by PDGF-BB. In conclusion, these results described a novel mechanism in atherosclerosis through PDGF-BB signaling in VSMC, in which MMP-2 expression is induced via extracellular signal-regulated kinases and p38 mitogen-activated protein kinase phosphorylation, as well as ROCK.
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Retracted Association of STAT4 gene polymorphism with systemic lupus erythematosus / lupus nephritis risk.
Nephrology (Carlton)
PUBLISHED: 04-11-2014
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The association of STAT4 gene polymorphism with systemic lupus erythematosus (SLE) / lupus nephritis (LN) results from the published studies is still conflicting. This meta-analysis was performed to evaluate the relationship between STAT4 rs7574865, rs16833431, rs11889341, rs8179673, rs10168266, rs7582694, rs3821236, rs7601754 gene polymorphism and SLE / LN, and to explore whether STAT4 gene polymorphism could become a predictive marker for SLE / LN risk.
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Gelatin-siloxane nanoparticles to deliver nitric oxide for vascular cell regulation: Synthesis, cytocompatibility, and cellular responses.
J Biomed Mater Res A
PUBLISHED: 04-10-2014
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Nitric oxide (NO) is an important mediator in cardiovascular system to regulate vascular tone and maintain tissue homeostasis. Its role in vascular cell regulation makes it promising to address the post-surgery restenosis problem. However, the application of NO is constrained by its high reactivity. Here, we developed a novel NO-releasing gelatin-siloxane nanoparticle (GS-NO NP) to deliver NO effectively for vascular cell regulation. Results showed that gelatin-siloxane nanoparticles (GS NPs) could be synthesized via sol-gel chemistry with a diameter of ?200 nm. It could be modified into GS-NO NPs via S-nitrosothiol (RSNO) modification. The synthesized GS-NO NPs could release a total of ?0.12 µmol/mg NO sustainably for 7 days following a first-order exponential profile. They showed not only excellent cytocompatibility, but also rapid intracellularization within 2 h. GS-NO NPs showed inhibition of human aortic smooth muscle cell (AoSMC) proliferation and promotion of human umbilical vein endothelial cell (HUVEC) proliferation in a dose-dependent manner, which is an important approach to prevent restenosis. With GS-NO NP dose at 100 µg/mL, the proliferation of AoSMCs could be slowed down whereas the growth of HUVECs was significantly promoted. We concluded that GS-NO NPs could have potential to be used as a promising nano-system to deliver NO for vascular cell regulation. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A:, 2014.
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Potential signal pathway of all-trans retinoic acid for MMP-2 and MMP-9 expression in injury podocyte induced by adriamycin.
J. Recept. Signal Transduct. Res.
PUBLISHED: 04-02-2014
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All-trans-retinoic acid (ATRA) can regulate some specific genes expression in various tissue and cells via nuclear retinoic acid receptors (RARs), including three subtypes: retinoic acid receptor-alpha (RAR-?), retinoic acid receptor-beta (RAR-?) and retinoic acid receptor-gamma (RAR-?). Podocyte injury plays a pivotal role in the progression of glomerulosclerosis (GS). This study was performed to study the potential signal pathway of ATRA in the expression of matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) in injury podocyte. Cells were divided into three groups: group of negative control (NC), group of injury podocyte induced by adriamycin (ADR) (AI) and group of ADR inducing podocyte injury model treated with ATRA (AA). The cells morphology changes were detected using microscope and scanning electron microscopy. MMP-2 and MMP-9 enzymic activity was detected using the gelatin zymography method. Protein and mRNA expressions of MMP-2, MMP-9, RAR-?, RAR-? and RAR-? were measured by western-blot and real-time RT-PCR. Enzymatic activity of MMP-2 and MMP-9 in group AA was significantly enhanced compared to AI group after ATRA-treated 24?h (p??0.05). RAR-? protein level was positively correlated with MMP-2 or MMP-9 protein expression (p?
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First report of Cryptosporidium spp. in white yaks in China.
Parasit Vectors
PUBLISHED: 03-29-2014
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Cryptosporidium is an enteric apicomplexan parasite, which can infect yaks, leading to reduction of milk production and poor weight gain. White yak (Bos grunniens) is a unique yak breed inhabiting only in Tianzhu Tibetan Autonomous County, Gansu province, northwestern China. The objective of the present study was to molecularly determine Cryptosporidium infection and species in white yaks.
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The changes of serum sKlotho and NGAL levels and their correlation in type 2 diabetes mellitus patients with different stages of urinary albumin.
Diabetes Res. Clin. Pract.
PUBLISHED: 02-27-2014
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To investigate the changes of serum anti-aging protein Klotho and neutrophil gelatinase-associated lipocalin (NGAL) levels and their correlation in type 2 diabetes mellitus (T2DM) patients at different stages of diabetic kidney disease (DKD) determined by urinary albuminuria.
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Ratio of n-3/n-6 PUFAs and risk of breast cancer: a meta-analysis of 274135 adult females from 11 independent prospective studies.
BMC Cancer
PUBLISHED: 02-18-2014
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Increased ratio of n-3/n-6 polyunsaturated fatty acids (PUFAs) in diet or serum may have a protective effect on the risk of breast cancer (BC); however, the conclusions from prospective studies are still controversial. The purpose of this study is to ascertain the relationship between intake ratio of n-3/n-6 PUFAs and the risk of BC, and estimate the potential summarized dose-response trend.
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Effects of Er-Zhi-Wan on microarchitecture and regulation of Wnt/?-catenin signaling pathway in alveolar bone of ovariectomized rats.
J. Huazhong Univ. Sci. Technol. Med. Sci.
PUBLISHED: 02-06-2014
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Recent studies have shown that Er-Zhi-Wan (EZW), a traditional Chinese medicine consisting of Herba Ecliptae (HE) and Fructus Ligustri Lucidi (FLL), had a definite antiosteoporotic effect on osteoporotic femur, but its effect on osteoporosis of alveolar bone remains unknown. In the present study, we investigated the effects of Er-Zhi-Wan (EZW) on the microarchitecture and the regulation of Wnt/?-catenin signaling pathway in the alveolar bone of ovariectomized rats. Thirty Sprague-Dawley rats were randomly divided into three groups: sham operation group (sham, n=10), ovariectomy (OVX) group (n=10), and OVX with EZW treatment group (EZW group, n=10). From one week after ovariectomy, EZW (100 mg/mL) or vehicle (distilled water) was fed (1 mL/100 g) once per day for 12 weeks until the sacrifice of the rats. The body weights were measured weekly. After sacrifice, the sera and mandible were collected and routinely prepared for the measurement of alveolar trabecular microarchitecture, serum levels of E2, bone-specific alkaline phosphatase (BALP) and tartrate-resistant acid phosphatase 5b (TRAP5b), as well as mandibular mRNA expression of Wnt/?-catenin signaling pathway molecules wnt3a, low-density lipoprotein receptor-related protein 5 (LRP5), ?-catenin and dickkopf homolog 1 (DKK1). The results showed that EZW treatment significantly prevented the body weight gain, degradation of alveolar trabecular microarchitecture and alveolar bone loss in the OVX rats. Furthermore, we observed that EZW could increase the serum levels of E2 and BALP, and decrease levels of serum TRAP5b in EZW group compared with vehicle group. In addition, RT-PCR results revealed that EZW upregulated the expression levels of wnt3a, LRP5 and ?-catenin, and reduced the expression of DKK1 in OVX rats. Taken together, our results suggested that EZW may have potential anti-osteoporotic effects on osteoporotic alveolar bone by stimulating Wnt/LRP5/?-catenin signaling pathway.
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Fumigaclavine C activates PPAR? pathway and attenuates atherogenesis in ApoE-deficient mice.
Atherosclerosis
PUBLISHED: 01-28-2014
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To develop alternative therapeutic strategy that reduces hypercholesterolemia, inflammation and atherosclerosis, we investigate if fumigaclavine C (FC), an indole alkaloid in structure, has anti-atherosclerosis function, and if so, what is the mechanism involved.
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Association of transforming growth factor-?1 T869C gene polymorphism with diabetic nephropathy risk.
Nephrology (Carlton)
PUBLISHED: 01-17-2014
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A possible association between the transforming growth factor-?1 (TGF-?1) T869C gene polymorphism and the risk of developing diabetic nephropathy (DN) remains unclear. This investigation was performed to assess if an association between the TGF-?1 T869C gene polymorphism and DN risk exists by using meta-analysis to combine comparable studies, thereby increasing sample size and statistical significance, and to identify patterns in various studies.
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Association of prohibitin-1 and 2 with oxidative stress in rats with renal interstitial fibrosis.
Mol. Biol. Rep.
PUBLISHED: 01-13-2014
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Prohibitins PHB1 and PHB2 are evolutionary conserved and pleiotropic proteins, which have been shown to be important factors in various cellular functions, including proliferation, tumour suppression, apoptosis, transcription, and mitochondrial protein folding. Recently, we demonstrated that down-regulation promoted renal interstitial fibrosis (RIF) in ureteral obstructed rats. Furthermore, the hypoxic conditions and oxidative stress have been implicated in obstruction-mediated renal disease. This study was performed to explore the association of PHBs with oxidative stress in a rat model of RIF. PHBs, the pro-fibrotic transforming growth factor-?1 (TGF-?1), and the extracellular matrix proteins collagen-IV (Col-IV) and fibronectin (FN) were evaluated, as were markers of oxidative stress [total reactive oxygen species (ROS), malondialdehyde (MDA)] and antioxidative capacity (superoxide dismutase, glutathione), and apoptosis. Our results showed a progressive increase in oxidative stress and concomitant decrease in antioxidants over a period of 4 weeks ureteral obstruction. Concomitantly, profibrotic components increased and PHB expression decreased. Overall, both PHBs were negatively correlated with the extent of observed fibrosis, TGF-?1, Col-IV, FN, ROS, MDA, and apoptosis.
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A novel approach for enriching cancer stem cells from the human SW-13 adrenocortical carcinoma cell line.
Anticancer Res.
PUBLISHED: 01-10-2014
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The present study was undertaken to develop a new method for enriching cancer stem cells (CSCs) from the human adrenal cortical carcinoma (ACC) cell line SW-13. Given that the existence of CSCs in ACC causes resistance to conventional chemotherapies, treatment with cyclophosphamide was used for in vivo selection of CSCs in a BALB/c nude mouse tumor xenograft model established using the ACC cell line SW-13. The characteristics of CSCs in three generations of tumor xenografts were assessed for single-cell colony formation, flat colony formation, and cell sphere formation in serum-free suspension culture. The formation rates of single-cell colonies, flat colonies, and cell spheres were significantly higher for tumor xenograft cells treated with cyclophosphamide than for untreated engrafted tumor cells. Flow cytometry to examine expression of the CSC markers C-X-C chemokine receptor type-4 (CXCR4; CD184) and ATP-binding cassette sub-family G member-2 (ABCG2; CDw338) revealed markedly higher levels of CXCR4 and ABCG2 in cyclophosphamide-treated xenograft tumor cells compared to untreated tumor cells. Together, these results indicate that cyclophosphamide treatment of tumor xenograft cells caused enrichment of CSCs with a strong capability for self-renewal and proliferation. In this method, the administration of cyclophosphamide selectively kills cancer cells without toxicity to CSCs and thereby provides a practical approach for achieving the enrichment of CSCs in ACC.
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p33(ING1b) methylation in fecal DNA as a molecular screening tool for colorectal cancer and precancerous lesions.
Oncol Lett
PUBLISHED: 01-09-2014
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The present study aimed to investigate the feasibility of detecting p33 inhibitor of growth 1b (p33(ING1b)) gene methylation in fecal DNA as a screening method for colorectal carcinoma (CRC) and precancerous lesions. The methylation of p33(ING1b) was analyzed in fecal samples from 61 patients with CRCs, 27 patients with precancerous lesions (advanced adenoma) and 20 normal individuals by nested methylation-specific polymerase chain reaction (nMSP) and fecal occult blood test. Methylated p33(ING1b) was detected in 73.77% of CRC patients and 62.96% of adenoma patients. By contrast, only 5% of normal individuals had methylated p33(ING1b). These results indicated 73.77% sensitivity for detecting CRC, 62.96% sensitivity for detecting precancerous lesions and 95% specificity of the assay for detecting CRCs and precancerous lesions. The detection of p33(ING1b) methylation status by incubation of DNA contained in agarose beads for bisulfite modification, followed by nMSP, is a promising non-invasive screening method for CRCs and precancerous lesions.
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Paradoxical expressions of natriuretic peptide receptor-C and neutral endopeptidase account for C-type natriuretic peptide decline during the progression of experimental obstructive nephropathy.
J Renin Angiotensin Aldosterone Syst
PUBLISHED: 11-06-2013
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C-type natriuretic peptide (CNP) selectively binds to the guanylyl cyclase coupled natriuretic peptide receptor (NPR)-B and exerts more potent antihypertrophic and antifibrotic properties. Elimination of CNP occurs mainly by neutral endopeptidase (NEP) and NPR-C.
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Association of peroxisome proliferator-activated receptors/retinoic acid receptors with renal diseases.
J. Recept. Signal Transduct. Res.
PUBLISHED: 09-19-2013
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Abstract Peroxisome proliferator-activated receptor-? (PPAR?), belongs to the nuclear receptor superfamily, and is a nuclear transcription receptor involving in the regulation of several biochemical pathways, such as cell growth, differentiation, and apoptosis. The nuclear retinoic acid receptors (RARs) are transcriptional transregulators that control the expression of specific subsets of genes in a ligand-dependent manner, and include three subtypes (RAR?, RAR?, and RAR?). These control the expression of specific gene subsets subsequent to ligand binding and to strictly control phosphorylation processes. The current status of knowledge indicates that there might be inter- or overlapping actions between PPAR? and RARs, and there might be an association of PPAR?/RARs with renal diseases. Various agonists of both receptor families seem to prevent or retard the progression of renal disease. Herein, we review if causal relationships can be established between PPAR?/RARs and renal diseases and its manifestations.
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Metal-free oxidative C(sp3)-H bond thiolation of ethers with disulfides.
Org. Lett.
PUBLISHED: 08-30-2013
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A novel method for the preparation of alkyl aryl sulfides through direct oxidation thiolation of commercial ethers with diaryl disulfides using di-tert-butyl peroxide (DTBP) as the oxidant without a metal catalyst was established. The C(sp(3))-H bond in various ethers was successfully converted into a C-S bond, and the corresponding sulfides were achieved with moderate to high yields.
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Biomimetic three-dimensional anisotropic geometries by uniaxial stretching of poly(?-caprolactone) films: Degradation and mesenchymal stem cell responses.
J Biomed Mater Res A
PUBLISHED: 06-25-2013
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Geometric cues have been used for a variety of cell regulation and tissue regenerative applications. While the function of geometric cues is being recognized, their stability and degradation behaviors are not well known. Here, we studied the influence of degradation on uniaxial-stretch-induced poly(?-caprolactone) (UX-PCL) ridge/groove arrays and further cellular responses. Results from accelerated hydrolysis in vitro showed that UX-PCL ridge/groove arrays followed a surface-controlled erosion, with an overall geometry remained even at ?45% film weight loss. Compared to unstretched PCL flat surfaces and/or ridge/groove arrays, UX-PCL ridge/groove arrays achieved an enhanced morphological stability against degradation. Over the degradation period, UX-PCL ridge/groove arrays exhibited an "S-shape" behavior of film weight loss, and retained more stable surface hydrophilicity and higher film mechanical properties than those of unstretched PCL surfaces. Human mesenchymal stem cells (MSCs) aligned better toward UX-PCL ridge/groove arrays when the geometries were remained intact, and became sensitive with gradually declined nucleus alignment and elongation to the geometric degradation of ridges. We speculate that uniaxial stretching confers UX-PCL ridge/groove arrays with enhanced stability against degradation in erosive environment. This study provides insights of how degradation influences geometric cues and further cell responses, and has implications for the design of biomaterials with stability-enhanced geometric cues for long-term tissue regeneration. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2013.
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A microporous metal-organic framework containing an exceptional four-connecting 4(2)6(4) topology and a combined effect for highly selective adsorption of CO2 over N2.
Dalton Trans
PUBLISHED: 05-17-2013
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Reported here is a new microporous metal-organic framework, namely [Zn(2)(L)(btc)(Hbtc)] [NH(2)(CH(3))(2)]·(DMF)(2)(H(2)O)(4) (1), which is synthesized solvo(hydro)thermally by the self-assembly of Zn(NO(3))(2), N(4),N(4)-di(pyridin-3-yl)-[1,1-biphenyl]-4,4-dicarboxamide (L) and 1,3,5-benzenetricarboxylate acid (H(3)btc). Its topology can be described as a four-connecting 4(2)6(4) matrix containing both tetrahedral metal and ligand nodes. Interestingly, such a matrix has the same topology symbol as that observed in the well known sodalite (SOD) net, but the td10 of 434 is different from the td10 of 791 for the SOD net, indicative of an exceptional four-connecting 4(2)6(4) net. Another outstanding point is the highly selective adsorption of CO(2) over N(2), possibly contributed by a combined effect from the charged skeleton, the existence of functional groups of -CONH- and -COOH in 1.
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Prohibitin is associated with antioxidative protection in hypoxia/reoxygenation-induced renal tubular epithelial cell injury.
Sci Rep
PUBLISHED: 05-15-2013
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Prohibitin is an evolutionary conserved and pleiotropic protein that has been implicated in various cellular functions, including proliferation, tumour suppression, apoptosis, transcription, and mitochondrial protein folding. We recently demonstrated that prohibitin downregulation results in increased renal interstitial fibrosis. Here we investigated the role of oxidative stress and prohibitin expression in a hypoxia/reoxygenation injury system in renal tubular epithelial cells with lentivirus-based delivery vectors to knockdown or overexpress prohibitin. Our results show that increased prohibitin expression was negatively correlated with reactive oxygen species, malon dialdehyde, transforming-growth-factor-?1, collagen-IV, fibronectin, and apoptosis (r = -0.895, -0.764, -0.798, -0.826, -0.817, -0.735; each P < 0.01), but positively correlated with superoxide dismutase, glutathione and mitochondrial membrane potential (r = 0.807, 0.815, 0.739; each P < 0.01). We postulate that prohibitin acts as a positive regulator of mechanisms that counteract oxidative stress and extracellular matrix accumulation and therefore has an antioxidative effect.
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Filter properties of chirped fiber Bragg grating Fabry-Perot cavity: a potential wavelength stabilizer of diode laser.
Appl Opt
PUBLISHED: 05-15-2013
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We investigate filter properties of chirped fiber Bragg grating (CFBG) (Fabry-Perot) F-P cavity through analyzing the coupled wave equation from one-dimensional Helmholtz equation. We derive an approximate formula of the reflectivity of a CFBG F-P cavity, simulate the central wavelength detuning, and calculate the central wavelength shift with the increase of ambient temperature. In the experiments, we measured the spectra of a diode laser with an FBG/CFBG F-P cavity at 0°C-110°C. The experimental results show that the CFBG F-P cavity can help a diode laser to obtain a less central wavelength shift and a narrower 3 dB reflection bandwidth, compared with the FBG F-P cavity at 0°C-110°C. The research results indicate that the CFBG F-P cavity is a potential wavelength stabilizer of uncooled diode laser.
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Anti-apoptotic proteins and catalase-dependent apoptosis resistance in nickel chloride-transformed human lung epithelial cells.
Int. J. Oncol.
PUBLISHED: 04-15-2013
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Chronic exposure to nickel compounds is associated with increased incidence of certain types of human cancer, including lung and nasal cancers. Despite intensive investigation, the oncogenic processes remain poorly understood. Apoptosis resistance is a key feature for tumor cells to escape physiological surveillance and acquire growth advantage over normal cells. Although NiCl2 exposure induces transformation of human lung epithelial cells, little information is available with regard to its molecular mechanisms, it is also not clear if the transformed cells are apoptosis resistant and tumorigenic. We explored the apoptosis resistance properties of nickel chloride?transformed human lung epithelial cells and the underlying mechanisms. The results showed that transformed BEAS-2B human lung epithelial cells are resistant to NiCl2-induced apoptosis. They have increased Bcl-2, Bcl-xL and catalase protein levels over the passage matched non?transformed counterparts. The mechanisms of apoptosis resistance are mitochondria?mediated and caspase-dependent. Forced overexpression of Bcl-2, Bcl-xL and catalase proteins reduced NiCl2-induced cell death; siRNA?mediated knockdown of their expression sensitized the cells to nickel-induced apoptosis, suggesting that Bcl-2, Bcl-xl and catalase protein expression plays a critical role in apoptosis resistance. Akt also participates in this process, as its overexpression increases Bcl-xL protein expression levels and attenuates NiCl2-induced apoptosis. Furthermore, transformed cells are tumorigenic in a xenograft model. Together, these results demonstrate that nickel-transformed cells are apoptosis?resistant and tumorigenic. Increased expression of Bcl-2, Bcl-xL and catalase proteins are important mechanisms contributing to transformed cell oncogenic properties.
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Carmichaeline A: a new C20-diterpenoid alkaloid from Aconitum carmichaeli.
Nat Prod Commun
PUBLISHED: 03-22-2013
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The roots of Aconitum carmichaeli Debx. are known for their medicinal value. A new C20-diterpenoid alkaloid designated as carmichaeline A (1) has been isolated, along with eight known diterpenoid alkaloids from the roots of the plant. Their structures were elucidated on the basis of spectroscopic data interpretation.
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[Effect of aldosterone on mesenteric resistance vessels in normal or heart failure rats and its mechanism].
Zhejiang Da Xue Xue Bao Yi Xue Ban
PUBLISHED: 03-19-2013
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To investigate the acute effects of aldosterone (ALD) on mesenteric resistance vessels in normal or heart failure (HF) rats and its mechanism.
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FSH modulates the expression of inhibin-alpha and the secretion of inhibins via orphan nuclear receptor NUR77 in ovarian granulosa cells.
Mol. Reprod. Dev.
PUBLISHED: 03-06-2013
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It has been previously reported that follicle-stimulating hormone (FSH) regulates the expression of inhibin-alpha in human granulosa cells, but the precise molecular pathway remains unknown. In the present study, we investigated the role of the orphan nuclear receptor, NUR77, in both the transcriptional regulation of the inhibin ?-subunit gene and the secretion of inhibins. Our results showed that in a human granulosa cell tumor-derived cell line (KGN) and in human granulosa-lutein cells (hGL), FSH induced the expression of NUR77 and inhibin-alpha, although inhibin-alpha expression did not increased following FSH treatment if NUR77 was knocked down. Furthermore, simply overexpressing or reducing NUR77 levels affected inhibin-alpha expression, while NUR77 overexpression improved the secretion of inhibin A and B from human granulosa cells. In addition, chromatin immunoprecipitation-PCR, avidin-biotin-conjugated DNA precipitation, and luciferase reporter assays confirmed that NUR77 directly regulated the transcription of the inhibin-alpha gene through the specific NGFI-B response element located within its promoter. In the ovarian granulosa cells of the Nur77 knockout mice, the mRNA levels of inhibin-alpha were decreased relative to wild-type mice. These data indicate a role of NUR77 in the regulation of inhibin-alpha in ovarian granulosa cells.
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Induction of apoptosis through caspase-independent or caspase-9-dependent pathway in mouse and human osteosarcoma cells by a new nitroxyl spin-labeled derivative of podophyllotoxin.
Apoptosis
PUBLISHED: 02-23-2013
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Previous study has found that a new nitroxyl spin-labeled derivative of podophyllotoxin, 4-[4"-(2",2",6",6"-tetramethyl-1"-piperidinyloxy)amino]-4-demethyl-epipodophyllotoxin (GP7), can induce apoptosis in human leukemia cells. However, there have been no studies about the effects of GP7 on osteosarcoma (OS) cells. Here, we observed the anti-OS effects of GP7 in mouse and human OS cells with the comparison of etoposide. GP7 and etoposide inhibited the proliferation of a panel of mouse and human OS cells in a concentration- or time-dependent manner, and the inhibitory effect of GP7 on the proliferation of mouse LM8 or human U2OS cells was 1.28- or 1.35-fold higher than that of etoposide. GP7 or etoposide augmented the anti-OS effects of methotrexate, adriamycin, cisplatin, or their combination, and the combined inhibitory effects of GP7 with MTX on the proliferation of LM8 cells was higher than those of etoposide with MTX. GP7 arrested the cell cycle in S phase but etoposide in G(2)/M phase. GP7 or etoposide induced sub-G(1) peak, apoptotic DNA fragmentation, activations of caspase-3, -8, -9, and DNA fragmentation factor, downregulation of Bcl-2 and Bcl-xL, upregulation of Bax and Bak, and cytochrome-c release from mitochondria in both mouse and human OS cells. GP7 or etoposide also induced endonuclease G translocation from mitochondria into cytosol in mouse cells. GP7- or etoposide-induced apoptotic DNA fragmentation of human OS cells was inhibited by the pan caspase inhibitor and caspase-9 inhibitor, not by caspase-8 inhibitor whereas it was not inhibited by the pan caspase inhibitor in mouse OS cells. Our findings indicate that GP7 is effective against mouse and human OS cells in vitro. The apoptotic DNA fragmentation in mouse OS cells may be mediated by caspase-independent pathway with the involvement of endonuclease G whereas in human OS cells by caspase-9-dependent pathway downstream of the cytochrome-c-initiated caspase cascade.
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Voxel-based diffusion tensor imaging of an APP/PS1 mouse model of Alzheimers disease.
Mol. Neurobiol.
PUBLISHED: 01-29-2013
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Increasing evidence has demonstrated that white matter (WM) disruptions, due to the injury of the axon and myelin, play an important role in the pathogenesis of Alzheimers disease (AD). Diffusion tensor imaging (DTI) is a sensitive modality to evaluate the WM integrity in both AD patients and animal models. In this study, an advanced DTI modality, employing a 7.0-T magnetic resonance imaging system, was used to analyze WM changes across the whole brain of an amyloid precursor protein/presenilin 1 (APP/PS1) mouse model. A voxel-based analysis was used to compare the quantitative DTI parameters automatically in both APP/PS1 mice (n?=?9) and wild-type (WT) controls (n?=?9). After DTI examination, the ultrastructure analysis was compared with DTI findings. Compared with WT controls, gray matter (GM) areas in APP/PS1 mice such as the cingulate cortex and the striatum showed significant fractional anisotropy (FA) and axial diffusivity (DA) increase, while the thalamus only showed a significant FA increase (p?
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9p21 polymorphisms increase the risk of peripheral artery disease in the Han Chinese population.
J. Int. Med. Res.
PUBLISHED: 01-23-2013
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A case-control study to investigate the association of the 9p21 single nucleotide polymorphisms (SNPs) rs10757274 and rs10757278 (known to be associated with coronary artery disease [CAD] risk) with peripheral arterial disease (PAD), in a Han Chinese population.
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Simplified stereological evaluation of renal morphology after unilateral ureteral obstruction.
Int J Med Sci
PUBLISHED: 01-23-2013
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Objective and methods: This study established a simple stereological method to obtain quantitative information about two- or three-dimensional structures based on observations from kidney sections in the unilateral ureteral obstruction(UUO) model. Results: Tubulointerstitial area(TA) and TA/the area of a rectangular field(RA) were raised gradually, but significantly, in the obstructed kidney from 1 to 3months post-ligation in comparison to the sham kidney of sham-operated rats(SOR). On the contrary, glomerular area(GA) and glomerular volume(GV) were decreased progressively over time, but significantly, in the obstructed kidney from 3weeks to 3months post-ligation compared to the sham kidney of SOR. UUO caused a progressive decline of TA and TA/RA in the contralateral kidney. More specifically, there were significant decreases in TA at 1,2,3months post-ligation, while in TA/RA only at 3months post-ligation in comparison to the right kidney of SOR. In contrast, GA and GV enhanced in a time-dependent manner in the contralateral kidney, in which the difference in GA reached significance only at 3months post-ligation, whereas the difference in GV reached significance from 1 to 3months post-ligation when comparing with the right kidney of SOR. Conclusions: Our results confirmed two typical features of obstructive nephropathy, including widen interstitial space and glomerular atrophy in the obstructed kidney, and compensatory growth of the contralateral kidney.
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Signaling pathways of prohibitin and its role in diseases.
J. Recept. Signal Transduct. Res.
PUBLISHED: 01-18-2013
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Prohibitin (PHB), appearing to be a negative regulator of cell proliferation and to be a tumor suppressor, has been connected to diverse cellular functions including cell cycle control, senescence, apoptosis and the regulation of mitochondrial activities. It is a growth regulatory gene that has pleiotropic functions in the nucleus, mitochondria and cytoplasmic compartments. However, in different tissues/cells, the expression of PHB was different, such as that it was increased in most of the cancers, but its expression was reduced in kidney diseases. Signaling pathways might be very important in the pathogenesis of diseases. This review was performed to provide a relatively complete signaling pathways flowchart for PHB to the investigators who were interested in the roles of PHB in the pathogenesis of diseases. Here, we review the signal transduction pathways of PHB and its role in the pathogenesis of diseases.
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Epithelial-mesenchymal transition during oncogenic transformation induced by hexavalent chromium involves reactive oxygen species-dependent mechanism in lung epithelial cells.
Toxicol. Appl. Pharmacol.
PUBLISHED: 01-13-2013
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Hexavalent chromium [Cr(VI)] is an important human carcinogen associated with pulmonary diseases and lung cancer. Exposure to Cr(VI) induces DNA damage, cell morphological change and malignant transformation in human lung epithelial cells. Despite extensive studies, the molecular mechanisms remain elusive, it is also not known if Cr(VI)-induced transformation might accompany with invasive properties to facilitate metastasis. We aimed to study Cr(VI)-induced epithelial-mesenchymal transition (EMT) and invasion during oncogenic transformation in lung epithelial cells. The results showed that Cr(VI) at low doses represses E-cadherin mRNA and protein expression, enhances mesenchymal marker vimentin expression and transforms the epithelial cell into fibroblastoid morphology. Cr(VI) also increases cell invasion and promotes colony formation. Further studies indicated that Cr(VI) uses multiple mechanisms to repress E-cadherin expression, including activation of E-cadherin repressors such as Slug, ZEB1, KLF8 and enhancement the binding of HDAC1 in E-cadherin gene promoter, but DNA methylation is not responsible for the loss of E-cadherin. Catalase reduces Cr(VI)-induced E-cadherin and vimentin protein expression, attenuates cell invasion in matrigel and colony formation on soft agar. These results demonstrate that exposure to a common human carcinogen, Cr(VI), induces EMT and invasion during oncogenic transformation in lung epithelial cells and implicate in cancer metastasis and prevention.
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In vivo quantitative whole-brain diffusion tensor imaging analysis of APP/PS1 transgenic mice using voxel-based and atlas-based methods.
Neuroradiology
PUBLISHED: 01-08-2013
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Diffusion tensor imaging (DTI) has been applied to characterize the pathological features of Alzheimers disease (AD) in a mouse model, although little is known about whether these features are structure specific. Voxel-based analysis (VBA) and atlas-based analysis (ABA) are good complementary tools for whole-brain DTI analysis. The purpose of this study was to identify the spatial localization of disease-related pathology in an AD mouse model.
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Biomimetic three-dimensional anisotropic geometries by uniaxial stretch of poly(?-caprolactone) films for mesenchymal stem cell proliferation, alignment, and myogenic differentiation.
Tissue Eng Part C Methods
PUBLISHED: 01-04-2013
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Anisotropic geometries are critical for eliciting cell alignment to dictate tissue microarchitectures and biological functions. Current fabrication techniques are complex and utilize toxic solvents, hampering their applications for translational research. Here, we present a novel simple, solvent-free, and reproducible method via uniaxial stretching for incorporating anisotropic topographies on bioresorbable films with ambitions to realize stem cell alignment control. Uniaxial stretching of poly(?-caprolactone) (PCL) films resulted in a three-dimensional micro-ridge/groove topography (inter-ridge-distance: ~6 ?m; ridge-length: ~90 ?m; ridge-depth: 200-900 nm) with uniform distribution and controllable orientation by the direction of stretch on the whole film surface. When stretch temperature (Ts) and draw ratio (DR) were increased, the inter-ridge-distance was reduced and ridge-length increased. Through modification of hydrolysis, increased surface hydrophilicity was achieved, while maintaining the morphology of PCL ridge/grooves. Upon seeding human mesenchymal stem cells (hMSCs) on uniaxial-stretched PCL (UX-PCL) films, aligned hMSC organization was obtained. Compared to unstretched films, hMSCs on UX-PCL had larger increase in cellular alignment (>85%) and elongation, without indication of cytotoxicity or reduction in cellular proliferation. This aligned hMSC organization was homogenous and stably maintained with controlled orientation along the ridges on the whole UX-PCL surface for over 2 weeks. Moreover, the hMSCs on UX-PCL had a higher level of myogenic genes expression than that on the unstretched films. We conclude that uniaxial stretching has potential in patterning film topography with anisotropic structures. The UX-PCL in conjunction with hMSCs could be used as "basic units" to create tissue constructs with microscale control of cellular alignment and elongation for tissue engineering applications.
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Prohibitin attenuates oxidative stress and extracellular matrix accumulation in renal interstitial fibrosis disease.
PLoS ONE
PUBLISHED: 01-01-2013
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Prohibitin is an evolutionary conserved and pleiotropic protein that has been implicated in various cellular functions, including proliferation, tumour suppression, apoptosis, transcription, and mitochondrial protein folding. Both prohibitin over- and under-expression have been implicated in various diseases and cell types. We recently demonstrated that prohibitin down-regulation results in increased renal interstitial fibrosis (RIF). Here we investigated the role of oxidative stress and prohibitin expression in RIF in unilateral ureteral obstructed rats. Lentivirus-based delivery vectors were used to knockdown or over-express prohibitin. Our results show that increased prohibitin expression was negatively correlated with the RIF index, reactive oxygen species, malon dialdehyde, transforming growth factor ?1, collagen IV, fibronectin, and cell apoptosis index. In conclusion, we postulate that prohibitin acts as a positive regulator of mechanisms that counteract oxidative stress and extracellular matrix accumulation and therefore has an antioxidative effect.
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Gross feature recognition of Anatomical Images based on Atlas grid (GAIA): Incorporating the local discrepancy between an atlas and a target image to capture the features of anatomic brain MRI.
Neuroimage Clin
PUBLISHED: 01-01-2013
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We aimed to develop a new method to convert T1-weighted brain MRIs to feature vectors, which could be used for content-based image retrieval (CBIR). To overcome the wide range of anatomical variability in clinical cases and the inconsistency of imaging protocols, we introduced the Gross feature recognition of Anatomical Images based on Atlas grid (GAIA), in which the local intensity alteration, caused by pathological (e.g., ischemia) or physiological (development and aging) intensity changes, as well as by atlas-image misregistration, is used to capture the anatomical features of target images. As a proof-of-concept, the GAIA was applied for pattern recognition of the neuroanatomical features of multiple stages of Alzheimers disease, Huntingtons disease, spinocerebellar ataxia type 6, and four subtypes of primary progressive aphasia. For each of these diseases, feature vectors based on a training dataset were applied to a test dataset to evaluate the accuracy of pattern recognition. The feature vectors extracted from the training dataset agreed well with the known pathological hallmarks of the selected neurodegenerative diseases. Overall, discriminant scores of the test images accurately categorized these test images to the correct disease categories. Images without typical disease-related anatomical features were misclassified. The proposed method is a promising method for image feature extraction based on disease-related anatomical features, which should enable users to submit a patient image and search past clinical cases with similar anatomical phenotypes.
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The prognostic role of BRAF mutation in metastatic colorectal cancer receiving anti-EGFR monoclonal antibodies: a meta-analysis.
PLoS ONE
PUBLISHED: 01-01-2013
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BRAF mutation has been investigated as a prognostic factor in metastatic colorectal cancer (mCRC) undergoing anti-EGFR monoclonal antibodies (moAbs), but current results are still inconclusive. The aim of this meta-analysis was to evaluate the relationship between BRAF mutation status and the prognosis of mCRC patients treated with moAbs.
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Genome-wide methylated DNA immunoprecipitation analysis of patients with polycystic ovary syndrome.
PLoS ONE
PUBLISHED: 01-01-2013
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Polycystic ovary syndrome (PCOS) is a complex, heterogeneous disorder of uncertain etiology. Recent studies suggested that insulin resistance (IR) plays an important role in the development of PCOS. In the current study, we aimed to investigate the molecular mechanism of IR in PCOS. We employed genome-wide methylated DNA immunoprecipitation (MeDIP) analysis to characterize genes that are differentially methylated in PCOS patients vs. healthy controls. Besides, we also identified the differentially methylated genes between patients with PCOS-non-insulin resistance (PCOS-NIR) and PCOS-insulin resistance (PCOS-IR). A total of 79 genes were differentially methylated between PCOS-NIR vs. PCOS-IR patients, and 40 genes were differentially methylated in PCOS patients vs. healthy controls. We analyzed these differentially methylated genes by constructing regulatory networks and protein-protein interaction (PPI) networks. Further, Gene Ontology (GO) and pathway enrichment analysis were also performed to investigate the biological functions of networks. We identified multiple categories of genes that were differentially methylated between PCOS-NIR and PCOS-IR patients, or between PCOS patients and healthy controls. Significantly, GO categories of immune response were differentially methylated in PCOS-IR vs. PCOS-NIR. Further, genes in cancer pathways were also differentially methylated in PCOS-NIR vs. PCOS-IR patients or in PCOS patients vs. healthy controls. The results of this current study will help to further understand the mechanism of PCOS.
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The myeloperoxidase-463 G>A polymorphism influences risk of colorectal cancer in southern China: a case-control study.
Asian Pac. J. Cancer Prev.
PUBLISHED: 12-01-2011
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Oxidative stress may be involved in colorectal carcinogenesis. Myeloperoxidase (MPO) is an endogenous oxidant enzyme that generates reactive oxygen species (ROS). We hypothesized that the MPO -463 locus polymorphism might therefore contribute to genetic susceptibility to colorectal adenomas.
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Expression of NLK and its potential effect in ovarian cancer chemotherapy.
Int. J. Gynecol. Cancer
PUBLISHED: 10-27-2011
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This study aimed to investigate the expression of NLK in ovarian cancer tissue and whether the proposed apoptosis induced by NLK was involved in ovarian cancer cells sensitive to cisplatin.
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Association of angiotensin converting enzyme insertion/deletion gene polymorphism with idiopathic nephrotic syndrome susceptibility in children: a meta-analysis.
J Renin Angiotensin Aldosterone Syst
PUBLISHED: 09-23-2011
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Angiotensin converting enzyme (ACE) gene contains either an insertion (I) allele or a deletion (D) allele forming three potential genotypes: II, ID and DD. The D allele or DD genotype has been reported to be associated with higher plasma ACE level. An assessment of the association between ACE I/D gene polymorphism and idiopathic nephrotic syndrome (INS) susceptibility in children is still controversial. This meta-analysis was performed to evaluate the association between ACE I/D gene polymorphism and the onset of INS.
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A meta-analysis of the association between angiotensin-converting enzyme insertion/deletion gene polymorphism and steroid-sensitive nephrotic syndrome in children.
J Renin Angiotensin Aldosterone Syst
PUBLISHED: 09-23-2011
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Angiotensin-converting enzyme (ACE) insertion/deletion (I/D) gene polymorphism correlates with circulating and cellular ACE concentration. Association between ACE I/D gene polymorphism and steroid-sensitive nephrotic syndrome (SSNS) risk in children is still controversial. This meta-analysis was performed to evaluate the relation between ACE I/D gene polymorphism and SSNS susceptibility in children.
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Sulfinylated azadecalins act as functional mimics of a pollen germination stimulant in Arabidopsis pistils.
Plant J.
PUBLISHED: 09-14-2011
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Polarized cell elongation is triggered by small molecule cues during development of diverse organisms. During plant reproduction, pollen interactions with the stigma result in the polar outgrowth of a pollen tube, which delivers sperm cells to the female gametophyte to effect double fertilization. In many plants, pistils stimulate pollen germination. However, in Arabidopsis, the effect of pistils on pollen germination and the pistil factors that stimulate pollen germination remain poorly characterized. Here, we demonstrate that stigma, style, and ovules in Arabidopsis pistils stimulate pollen germination. We isolated an Arabidopsis pistil extract fraction that stimulates Arabidopsis pollen germination, and employed ultra-high resolution electrospray ionization (ESI), Fourier-transform ion cyclotron resonance (FT-ICR) and MS/MS techniques to accurately determine the mass (202.126?Da) of a compound that is specifically present in this pistil extract fraction. Using the molecular formula (C10H19NOS) and tandem mass spectral fragmentation patterns of the m/z (mass to charge ratio) 202.126 ion, we postulated chemical structures, devised protocols, synthesized N-methanesulfinyl 1- and 2-azadecalins that are close structural mimics of the m/z 202.126 ion, and showed that they are sufficient to stimulate Arabidopsis pollen germination in vitro (30??m stimulated approximately 50% germination) and elicit accession-specific response. Although N-methanesulfinyl 2-azadecalin stimulated pollen germination in three species of Lineage I of Brassicaceae, it did not induce a germination response in Sisymbrium irio (Lineage II of Brassicaceae) and tobacco, indicating that activity of the compound is not random. Our results show that Arabidopsis pistils promote germination by producing azadecalin-like molecules to ensure rapid fertilization by the appropriate pollen.
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[Combination of AD5-10 and epirubicin in treating rheumatoid arthritis].
Zhongguo Yi Xue Ke Xue Yuan Xue Bao
PUBLISHED: 09-13-2011
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To investigate the mechanism of anti-death receptor 5-10 (AD5-10) combined with epirubicin in treating rheumatoid arthritis (RA).
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1,4-Bis(1H-benzimidazol-1-yl)benzene.
Acta Crystallogr Sect E Struct Rep Online
PUBLISHED: 07-28-2011
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In the title compound, C(20)H(14)N(4), the dihedral angles between the central benzene ring and the pendant benzimidazole ring systems are 46.60?(15) and 47.89?(16)°. The dihedral angle between the benzimidazole ring systems is 85.62?(12)° and the N atoms lie to the same side of the mol-ecule. In the crystal, mol-ecules are linked by C-H?N inter-actions and weak aromatic ?-? stacking [shortest centroid-centroid separation = 3.770?(2)?Å] is observed.
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7-Amino-4-hy-droxy-4-trifluoro-methyl-3,4-dihydro-quinolin-2(1H)-one.
Acta Crystallogr Sect E Struct Rep Online
PUBLISHED: 07-27-2011
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The title compound, C(10)H(9)F(3)N(2)O(2), was prepared by the reaction of m-phenyl-enediamine and ethyl 4,4,4-trifluoro-acetoacetate. In the crystal, inter-molecular C-H? F, N-H?F, O-H?N and N-H?O inter-actions contribute to the crystal packing.
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Insertion/deletion (I/D) polymorphism of angiotensin-converting enzyme gene in steroid-resistant nephrotic syndrome for children: a genetic association study and meta-analysis.
Ren Fail
PUBLISHED: 07-27-2011
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An assessment of the association of angiotensin-converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism with steroid-resistant nephrotic syndrome (SRNS) risk in children is still controversial. A meta-analysis was performed to evaluate the relation between ACE gene polymorphisms and SRNS susceptibility. The relevant studies were screened from electronic database and eligible investigations were synthesized using meta-analysis methods. Seven investigations were identified for the analysis of association between ACE I/D gene polymorphism and SRNS risk in children, including five in Asians, one in Caucasians, and one in Africans. There was not a markedly positive association between D allele or DD genotype and SRNS susceptibility in Asians (OR = 1.60, p = 0.26; OR = 1.90, p = 0.38) and for Caucasian population (OR = 0.92, p = 0.86; OR = 0.27, p = 0.22). However, an association of D allele with SRNS susceptibility was observed (OR = 4.67, p = 0.003) in Africans, but not for DD genotype (OR = 6.00, p = 0.05). Interestingly, II genotype seemed to play a positive role against SRNS onset for Asians and African children (OR = 0.51, p = 0.02; OR = 0.07, p = 0.02), but not for Caucasians (OR = 0.33, p = 0.30). In conclusion, our results indicate that D allele or DD homozygous might not be a significant genetic molecular marker for the development of SRNS in Asians and Caucasian children. However, D allele seemed be associated with SRNS risk for Africans but DD genotype did not.
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Numerical and experimental study of the thermal stress of silicon induced by a millisecond laser.
Appl Opt
PUBLISHED: 07-21-2011
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A spatial axisymmetric finite element model of single-crystal silicon irradiated by a 1064 nm millisecond laser is used to investigate the thermal stress damage induced by a millisecond laser. The transient temperature field and the thermal stress field for 2 ms laser irradiation with a laser fluence of 254 J/cm(2) are obtained. The numerical simulation results indicate that the hoop stresses along the r axis on the front surface are compressive stress within the laser spot and convert to tensile stress outside the laser spot, while the radial stresses along the r axis on the front surface and on the z axis are compressive stress. The temperature of the irradiated center is the highest temperature obtained, yet the stress is not always highest during laser irradiation. At the end of the laser irradiation, the maximal hoop stress is located at r=0.5 mm and the maximal radial stress is located at r=0.76 mm. The temperature measurement experiments are performed by IR pyrometer. The numerical result of the temperature field is consistent with the experimental result. The damage morphologies of silicon under the action of a 254 J/cm(2) laser are inspected by optical microscope. The cracks are observed initiating at r=0.5 mm and extending along the radial direction.
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[In vivo high-resolution imaging of rat retina with optical coherence tomography and the expression of Bcl-2, Bax, Caspase-3 mRNA during critical period plasticity].
Zhonghua Yan Ke Za Zhi
PUBLISHED: 07-16-2011
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To observe the changes of retinal thickness during critical period plasticity in rat, and to investigate whether apoptosis participates in the structural forming of retina.
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Temperature field analysis of single layer TiO2 film components induced by long-pulse and short-pulse lasers.
Appl Opt
PUBLISHED: 07-12-2011
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To study the differences between the damaging of thin film components induced by long-pulse and short-pulse lasers, a model of single layer TiO(2) film components with platinum high-absorptance inclusions was established. The temperature rises of TiO(2) films with inclusions of different sizes and different depths induced by a 1 ms long-pulse and a 10 ns short-pulse lasers were analyzed based on temperature field theory. The results show that there is a radius range of inclusions that corresponds to high temperature rises. Short-pulse lasers are more sensitive to high-absorptance inclusions and long-pulse lasers are more easily damage the substrate. The first-damage decision method is drawn from calculations.
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Variable frequencies of apolipoprotein e genotypes and its effect on serum lipids in the guangxi zhuang and han children.
Int J Mol Sci
PUBLISHED: 07-11-2011
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Guangxi Zhuang, the largest ethnic minority in China, is located in the southern part of the country, and well-known to the world as the longevity village. Studies of apolipoprotein E (APOE) polymorphism in adults suggest the lower frequencies of E4 allele and E4/E4 genotype may account, in part, for the favorable lipid profiles of Guangxi Zhuang. However, the effect of APOE polymorphism on serum lipids in the Guangxi Zhuang children is yet unknown to date. In the present study, genomic DNA was extracted from 278 Guangxi Zhuang and 200 Guangxi Han children. APOE genotypes were determined by PCR-restriction fragment length polymorphism (RFLP) analysis. The fasting serum lipoprotein a [Lp(a)], total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein A1 (apoA1) and apoB were measured. Our results demonstrated that no significant differences in serum lipids were observed between the Guangxi Zhuang and Han children. The E4/E4 and E4/E3 genotypic frequencies were significantly lower in the Guangxi Zhuang children compared with the Guangxi Han children, whereas for E2/E2, E3/E2 and E4/E2 genotypic frequencies the opposite was presented. Though no significant differences in serum lipid concentrations were found for variant alleles both in the Guangxi Zhuang and Han children, the trend was observed in the association of higher levels of Lp(a), TC, TG and LDL-C with E4 allele in the Guangxi Zhuang children. In conclusion, a significant heterogeneity in APOE genetic variation indeed exists between the Guangxi Zhuang and Han ethnic group. The E4 allele may serve as a genetic marker for susceptibility to higher lipid profiles in the Guangxi Zhuang children. Lifestyle should be modified, according to APOE polymorphism even in the young children.
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All-trans retinoic acid can regulate the expressions of gelatinases and apolipoprotein E in glomerulosclerosis rats.
Vascul. Pharmacol.
PUBLISHED: 06-17-2011
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Apolipoprotein E (apoE) is an important plasma protein in cholesterol homeostasis and plays a key role in the pathogenesis of glomerulosclerosis (GS). Gelatinases include matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9). The abnormal expressions of gelatinases are implicated in the pathogenesis of extracellular matrix accumulation. All-trans retinoic acid (ATRA) is an import biological agent which can play a protective role against GS. We performed this investigation to explore whether ATRA could regulate the expressions of gelatinases and apoE in the glomerulus of GS rats. 120 Wistar rats were randomly divided into three groups: sham operation group (SHO), glomerulosclerosis model group without treatment (GS) and GS model group treated with ATRA (GA). The GS disease was established by uninephrectomy and adriamycin injection. At the end of 9 and 13 weeks, the relevant samples were collected and determined. Compared with GS group at 9/13 weeks, values of 24-hour urine total protein, 24-hour urine excretion for albumin, blood urea nitrogen, serum creatinine and glomerulosclerosis index, and protein expressions of apoE, transforming growth factor-?l (TGF-?1), ?-smooth muscle actin, collagen-IV and fibronectin in glomerulus and mRNA expressions of apoE and TGF-?1 in renal tissue were significantly down-regulated by ATRA (each P<0.01). However, the expressions of MMP-2 and MMP-9 (mRNA, protein and activity) were enhanced in GA group than those in GS group. In conclusion, gelatinases are associated with apoE expression, and ATRA can increase the gelatinases expressions and reduce the accumulation of apoE in glomerulus of GS rats, but the detailed mechanism needs to be elucidated in the future.
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The association between angiotensin-converting enzyme insertion/deletion gene variant and risk of focal segmental glomerulosclerosis: a systematic review and meta-analysis.
J Renin Angiotensin Aldosterone Syst
PUBLISHED: 06-07-2011
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The association of the angiotensin-converting enzyme (ACE) insertion/deletion (I/D) gene polymorphism with the risk of focal segmental glomerulosclerosis (FSGS) is still controversial. A meta-analysis was performed to evaluate the association between ACE I/D gene polymorphism and FSGS susceptibility.
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Modulation of vitamin D signaling is a potential therapeutic target to lower cardiovascular risk in chronic kidney disease.
Med. Sci. Monit.
PUBLISHED: 06-02-2011
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While it is true that many traditional cardiovascular risk factors are amenable to intervention in chronic kidney disease (CKD), the results of intervention may not be as efficacious as those obtained in the general population. Thus, there may also be a unique milieu established in CKD, which causes excess cardiovascular disease (CVD) burden by mechanisms that are as yet not fully recognized. Recently, vitamin D has sparked widespread interest because of its potential favorable benefits on CVD. However, the mechanisms for how vitamin D may improve CVD risk markers and outcomes have not been fully elucidated. Furthermore, hypovitaminosis D is highly prevalent in the CKD cohort. Given this background, we hypothesize that low vitamin D status may act as a new CVD risk marker, and modulation of vitamin D signaling may be a potential therapeutic target to lower cardiovascular risk in CKD. The data presented in this review support that the low vitamin D status may be linked with the high cardiovascular risk in CKD, based on both the biological effects of vitamin D itself on the cardiovascular system, and the cross-actions between vitamin D signaling and the multiple metabolic pathways. Considering the high prevalence of hypovitaminosis D, limited natural vitamin D food sources, and reduced sun exposure in CKD patients, recommendations for treatment of hypovitaminosis D mainly focus on exogenous supplementation with vitamin D and its analogues. Although promising, when to start therapy, the route of administration, the dose, and the duration remain need to be discussed.
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