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Objective: To investigate the lipid metabolic effect and mechanism of water extract of lotus leaves(Traditional Chinese Medicine)., Methods: Isolated SD rat lipid tissues were suspended in organ baths containing Krebs solution, and the effect of lotus leaf water extract on free fatty acids (FFA) release was observed; The experimental obesity rat model was established by feeding them high glucose and fat diets, then the changes of body weight and blood lipid were measured in the model rats after intragastric administration with water extract of lotus leaves for four weeks, and the expressions of peroxisome proliferator-activated receptor gamma (PPAR-γ) and leptin were examined by RT-PCR and immunohistochemical., Results: The ex vivo experiment showed that water extract of lotus leaves effectively promoted the FFA release from isolated lipid tissues. In vivo experiment, similarly to Orlistat, water extract of lotus leaves(60 mg/kg)markedly decreased the body weight and blood lipid of experimental obesity rats( P <0.05), and obviously reduced the expressions of PPAR-γ and leptin( P <0.05)., Conclusions: Water extract of lotus leaves greatly improves the expression of PPAR-γ and leptin, which can promote the lipid mobilization and dissolution, reduce the body weight and blood lipid of adult rats with experimental obesity, therefore is expected to be developed into lipid-lowering diet pills.

Our previous study showed that RalA-binding protein 1 (RLIP76) is overexpressed in gliomas and is associated with higher tumour grade and decreased patient survival. Furthermore, RLIP76 downregulation increases chemosensitivity of glioma cells to temozolomide by inducing apoptosis. However, other mechanisms underlying RLIP76-associated chemoresistance are unknown. In this study, we investigated the effect of RLIP76 depletion on autophagy. RLIP76 was knocked down in U251 glioma cells using shRNA and autophagy-related proteins, and PI3K/Akt signalling components were evaluated. RLIP76 depletion significantly increased cell autophagy as demonstrated by a significant increase in LC3 II, autophagy protein 5 (ATG-5), and Beclin1, and a decrease in p62 expression levels. Furthermore, RLIP76 knockdown increased autophagic flux in U251 cells as autolysosome numbers increased relative to autophagosome numbers. Autophagy induced by RLIP76 knockdown resulted in increased apoptosis that was independent of temozolomide treatment. Moreover, RLIP76 knockdown decreased PI3K and Akt activation. RLIP76 depletion also resulted in decreased levels of the anti-apoptotic protein Bcl2. LY294002, a PI3K/Akt pathway inhibitor, led to increased autophagy and apoptosis in U251 RLIP76-depleted cells. Therefore, RLIP76 knockdown increased autophagic flux and apoptosis in U251 glioma cells, possibly through inhibition of the PI3K/Akt pathway. Thus, this study provides a novel mechanism for the role of RLIP76 in glioma pathogenesis and chemoresistance.

Background: Antihypertensive drug use is inconsistently associated with the risk of dementia, Alzheimer's disease, cognitive impairment, and cognitive decline. Therefore, we conducted a meta-analysis of available prospective cohort studies to summarize the evidence on the strength of these relationships., Methods: Three electronic databases including MedLine, Embase, and the Cochrane Library were searched to identify studies from inception to April 2017. Only prospective cohort studies that reported effect estimates with corresponding 95% confidence intervals (CIs) of dementia, Alzheimer's disease, cognitive impairment, and cognitive decline for antihypertensive drug use versus not using antihypertensive drugs were included., Results: We included 10 prospective cohort studies reporting data on 30,895 individuals. Overall, participants who received antihypertensive drugs had lower incidence of dementia (relative risk [RR]: 0.86; 95% CI: 0.75-0.99; p = 0.033), while there was no significant effect on the incidence of Alzheimer's disease (RR: 0.83; 95% CI: 0.64-1.09; p = 0.154), cognitive impairment (RR: 0.89; 95% CI: 0.57-1.38; p = 0.596), and cognitive decline (RR: 1.11; 95% CI: 0.86-1.43; p = 0.415). Further, the incidence of Alzheimer's disease might be affected by antihypertensive drug use in participants with specific characteristics., Conclusions: Antihypertensive drug use was associated with a significantly reduced risk of dementia, but not with the risk of Alzheimer's disease, cognitive impairment, and cognitive decline.

Background: In bacteria, transcription units can be insulated by placing a terminator in front of a promoter. In this way promoter leakage due to the read-through from an upstream gene or RNA polymerase unspecific binding to the DNA is, in principle, removed. Differently from bacterial terminators, yeast S. cerevisiae terminators contain a hexamer sequence, the efficiency element, that strongly resembles the eukaryotic TATA box i.e. the promoter sequence recognized and bound by RNA polymerase II., Results: By placing different yeast terminators (natural and synthetic) in front of the CYC1 yeast constitutive promoter stripped of every upstream activating sequences and TATA boxes, we verified that the efficiency element is able to bind RNA polymerase II, hence working as a TATA box. Moreover, terminators put in front of strong and medium-strength constitutive yeast promoters cause a non-negligible decrease in the promoter transcriptional activity., Conclusions: Our data suggests that RNA polymerase II molecules upon binding the insulator efficiency element interfere with protein expression by competing either with activator proteins at the promoter enhancers or other RNA polymerase II molecules targeting the TATA box. Hence, it seems preferable to avoid the insulation of non-weak promoters when building synthetic gene circuit in yeast S. cerevisiae .

Age-related hearing loss (AHL) is one of the most common sensory disorders among elderly persons. The inwardly rectifying potassium channel 5.1 (Kir5.1) plays a vital role in regulating cochlear K(+) circulation which is necessary for normal hearing. The distribution of Kir5.1 in C57BL/6J mice cochleae, and the relationship between the expression of Kir5.1 and the etiology of AHL were investigated. Forty C57BL/6J mice were randomly divided into four groups at 4, 12, 24 and 52 weeks of age respectively. The location of Kir5.1 was detected by immunofluorescence technique. The mRNA and protein expression of Kir5.1 was evaluated in mice cochleae using real-time polymerase-chain reactions (RT-PCR) and Western blotting respectively. Kir5.1 was detected in the type II and IV fibrocytes of the spiral ligament in the cochlear lateral wall of C57BL/6J mice. The expression levels of Kir5.1 mRNA and protein in the cochleae of aging C57BL/6J mice were down-regulated. It was suggested that the age-related decreased expression of Kir5.1 in the lateral wall of C57BL/6J mice was associated with hearing loss. Our results indicated that Kir5.1 may play an important role in the pathogenesis of AHL.

Water stress causes considerable yield losses in sugarcane. To investigate differentially expressed genes under water stress, a pot experiment was performed with the sugarcane variety GT21 at three water-deficit levels (mild, moderate, and severe) during the elongation stage and gene expression was analyzed using microarray technology. Physiological parameters of sugarcane showed significant alterations in response to drought stress. Based on the expression profile of 15,593 sugarcane genes, 1,501 (9.6%) genes were differentially expressed under different water-level treatments; 821 genes were upregulated and 680 genes were downregulated. A gene similarity analysis showed that approximately 62.6% of the differentially expressed genes shared homology with functional proteins. In a Gene Ontology (GO) analysis, 901 differentially expressed genes were assigned to 36 GO categories. Moreover, 325 differentially expressed genes were classified into 101 pathway categories involved in various processes, such as the biosynthesis of secondary metabolites, ribosomes, carbon metabolism, etc. In addition, some unannotated genes were detected; these may provide a basis for studies of water-deficit tolerance. The reliability of the observed expression patterns was confirmed by RT-PCR. The results of this study may help identify useful genes for improving drought tolerance in sugarcane.

Purpose . To use in vitro and in vivo models to evaluate Glechoma longituba extract to provide scientific evidence for this extract's antiurolithic activity. Materials and Methods . Potassium citrate was used as a positive control group. Oxidative stress (OS) markers and the expression of osteopontin (OPN) and kidney injury molecule-1 (KIM-1) were measured to assess the protective effects of Glechoma longituba . Multiple urolithiasis-related biochemical parameters were evaluated in urine and serum. Kidneys were harvested for histological examination and the assessment of crystal deposits. Results . In vitro and in vivo experiments demonstrated that treatment with Glechoma longituba extract significantly decreased calcium oxalate- (CaOx-) induced OPN expression, KIM-1 expression, and OS compared with the positive control group ( P < 0.05). Additionally, in vivo rats that received Glechoma longituba extract exhibited significantly decreased CaOx deposits and pathological alterations ( P < 0.05) compared with urolithic rats. Significantly lower levels of oxalate, creatinine, and urea and increased citrate levels were observed among rats that received Glechoma longituba ( P < 0.05) compared with urolithic rats. Conclusion . Glechoma longituba has antiurolithic effects due to its possible combined effects of increasing antioxidant levels, decreasing urinary stone-forming constituents and urolithiasis-related protein expression, and elevating urinary citrate levels.

Background: Dysregulated endocytosis of membrane proteins contributes significantly to several hallmarks of cancer. Basigin can enhance cancer progression, but its precise mechanism remains unclear. CD98 promotes cell spreading and tumorigenicity by triggering integrin clustering and enhancing cell adhesion to the extracellular matrix. The endocytosis and recyle of basigin and CD98 might play critical roles in cancer., Methods: The role of CD98 was confirmed in liver cancer cells by cell spreading in vitro and tumorigenicity by nude mice xenograft tumor assay in vivo; membrane expression of basigin and CD98 in SMMC-7721 was measured by FCAS; pull down and SPR analysis were uses to reveal the direct association between basigin and CD98; DsRed1 tagged CD98 was blocked in the cytoplasm in K7721 (whose basigin was knockn out) and had a well colocalization with ER and Rab5a positive recycling endosomes under co-focal; finally, by FRET imaging and FCAS we observed the internalization of basigin and CD98 was flotillin-1-regulated, and their recycle at early steps was Arf6-mediated., Results: Basigin and CD98 were highly expressed and co-localized on the human hepatocellular carcinoma (HCC) cell membrane; basigin can directly bind to CD98, mediating CD98 redistribution on the HCC cell membrane and activating the downstream integrin signaling pathway. Internalization of basigin and CD98 was flotillin-1 regulated the and their recycling was mediated by Arf6. This recycling process for basigin and CD98 promotes cell spreading and tumor growth in liver cancer xenografts., Conclusion: Basigin, as a redistribution chaperone of CD98, plays a critical role in promoting cell spreading and the progression of hepatocellular carcinoma.

Mouse models are widely used for studying gastrointestinal (GI) tract-related diseases. It is necessary and important to develop a new set of primers to monitor the mouse gut microbiota. In this study, 16S rRNA gene-targeted group-specific primers for Firmicutes, Actinobacteria, Bacteroidetes, Deferribacteres, "Candidatus Saccharibacteria," Verrucomicrobia, Tenericutes, and Proteobacteria were designed and validated for quantification of the predominant bacterial species in mouse feces by real-time PCR. After confirmation of their accuracy and specificity by high-throughput sequencing technologies, these primers were applied to quantify the changes in the fecal samples from a trinitrobenzene sulfonic acid-induced colitis mouse model. Our results showed that this approach efficiently predicted the occurrence of colitis, such as spontaneous chronic inflammatory bowel disease in transgenic mice. The set of primers developed in this study provides a simple and affordable method to monitor changes in the intestinal microbiota at the phylum level., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Objective: To extract two kinds of phenols 4-hydroxy-3, 5-dimethoxy-4-(2-oxopropyl) cyclohexa-2, 5-dien-l-one and 6-methoxy-5,7-dihydroxy coumarin (named as I and H compounds respectively) from Ajania salicifolia and to investigate their antioxidation and cytotoxicity to tumors and explore their pro-apoptosis mechanism., Methods: The antioxidant activities of two compounds were assessed by ABTS and DPPH radical-scavenging assays. Two compounds were evaluated for their cytotoxicity against human chronic myelogenous leukemia (K562) cells using the MIT assay. The expression of NF-kappaB P65 mRNA in K562 apoptotic cells was measured by reverse transcription-polymerase chain reaction (RT-PCR), real-time quantitative PCR. In addition, protein expression levels of the NF-ICB P65, p-Akt, Fas, P-catenina and E-cadherin were also measured by Western blot., Results: (1) We found that compound I displayed significant inoxidizability, while compound II had no obvious antioxidizability. (2) In cytotoxicity experiments, compound I didn't display cytotoxicity while compound H displayed obvious cytotoxicity. (3) Compared with the blank group, the expression of NF-kappaB P65 mRNA in K562 cell after treatment with compound II was obviously up-regulated. (4) Compared with the blank group, the expression levels of NF-kappaB P65, Fas, beta-catenina and E-cadherin were significantly increased in compound II treated groups and it appeared obvious dose-effect relationship between the expression of protein and drug concentration., Conclusion: Two phenols have obvious antioxidizability and cytotoxicity respectively. On the one hand, the tumor-suppressing mechanism of compound II maybe act by up-regulation the expression of NF-kappaB P65 and Fas protein; thereby, affecting the classical Fas apoptosis signaling pathways. On the other hand, it can also up-regulate the expression of protein beta-catenin and E-cadherin, which participate in the adhesion between cells, and accordingly, playing an important role in preventing the proliferation and metastasis of cancer cells.

Objective: To understand the epidemic situation of schistosomiasis in 27 counties (cities, districts) that reached the criteria of schistosomiasis infection controlled in Anhui Province., Methods: According to the requirement of The National Assessment Scheme of Schistosomiasis, 81 administrative villages where the schistosomiasis epidemic situation was relatively heaver in above-mentioned 27 counties (1 village per town, 3 towns per county) were sampled and investigated., Results: From 2012 to 2014, 81 villages were investigated, and 34,293 residents received the serum examinations, and 1,086 were positive with a positive rate of 3.17% (0.65%-9.58%), and the positives received stool examinations and the average stool positive rate was 0.37% (0-4.0%). The calculated prevalence of human infection was 0.01%. A total of 3 057 domestic animals were investigated including 438 cattle, 2,550 sheep, and 69 other animals, and no infections were detected. A total of 11,261 living Oncomelania hupensis snails were collected and detected, but no schistosome infected snails were found. Before this investigation, no infected snails were detected for more than 2 years [average 2.3 (2-6) years], and no acute schistosome infection cases were found for more than 2 years [average 4.9 (2-9) years]., Conclusion: The infection rates of schistosomiasis in residents and domestic animals are relatively low, and no schistosome infected snails are found in the regions.

The potential effects of Fa extract on the prevention and treatment of CaOx nephrolithiasis were analyzed in an ethylene glycol- (EG-) induced CaOx crystallization model in rats and an in vitro assay. Multiple biochemical variables were measured in the urine and kidney. Kidney sections were subjected to histopathological and immunohistochemical analyses. Urolithiasis-related osteopontin (OPN) was evaluated by Western blotting. The in vitro assay revealed the significant inhibition of crystal formation (3.50 ± 1.43) and dilution of formed crystals (12.20 ± 3.35) in the group treated with 1 mg/mL Fa extract compared with the control group (52.30 ± 4.71 and 53.00 ± 4.54, resp.) (p < 0.05). The in vivo experiments showed that prophylactic treatment with Fa aqueous extract significantly prevented EG-induced renal crystallization and pathological alterations compared with nephrolithic rats (p < 0.05). Significantly lower levels of oxidative stress, oxalate, and OPN expression as well as increased citrate and urine output levels were observed in both the low- and high-dose prophylactic groups (p < 0.05). However, in the low- and high-dose therapeutic groups, none of these indexes were significantly improved (p > 0.05) except for urinary oxalate in the high-dose therapeutic groups (p < 0.05). Fa extract prevented CaOx crystallization and promoted crystal dissolution in vitro. Additionally, it was efficacious in preventing the formation of CaOx nephrolithiasis in rats.

Focal adhesions (FAs), integrin-mediated macromolecular complexes located at the cell membrane extracellular interface, have been shown to regulate cell adhesion and migration. Our previous studies have indicated that HAb18G/CD147 (CD147) is involved in cytoskeleton reorganization and FA formation in human hepatocellular carcinoma (HCC) cells. However, the precise mechanisms underlying these processes remain unclear. In the current study, we determined that CD147 was involved in vinculin-mediated FA focal adhesion formation in HCC cells. We also found that deletion of CD147 led to reduced vinculin-mediated FA areas (P<0.0001), length/width ratios (P<0.0001), and mean intensities (P<0.0001). CD147 promoted lamellipodia formation by localizing Arp2/3 to the leading edge of the cell. Deletion of CD147 significantly reduced the fluorescence (t1/2) recovery times (22.7±3.3 s) of vinculin-mediated focal adhesions (P<0.0001). In cell-spreading assays, CD147 was found to be essential for dynamic focal adhesion enlargement and disassembly. Furthermore, the current data showed that CD147 reduced tyrosine phosphorylation in vinculin-mediated focal adhesions, and enhanced the accumulation of the acidic phospholipid phosphatidylinositol-4, 5-bisphosphate (PIP2). Together, these results revealed that CD147 is involved in vinculin-mediated focal adhesion formation, which subsequently promotes cytoskeleton reorganization to facilitate invasion and migration of human HCC cells.

1,3,4-Oxadiazole derivatives have drawn continuing interest over the years because of their varied biological activities. In order to search for novel anticancer agents, we designed and synthesized a series of new 1,3,4-oxadiazole derivatives containing benzotriazole moiety as potential focal adhesion kinase (FAK) inhibitors. All the synthesized compounds were firstly reported. Among the compounds, compound 4 shows the most potent inhibitory activity against MCF-7 and HT29 cell lines with IC50 values of 5.68 μg/ml and 10.21 μg/ml, respectively. Besides, all the compounds were assayed for FAK inhibitory activity using the TRAP-PCR-ELISA assay. The results showed compound 4 exhibited the most potent FAK inhibitory activity with IC50 values of 1.2±0.3 μM. Docking simulation by positioning compound 4 into the FAK structure active site was performed to explore the possible binding mode. Apoptosis which was analyzed by flow cytometry, demonstrated that compound 4 induced apoptosis against MCF-7 cells. Therefore, compound 4 may be a potential anticancer agent against MCF-7 cancer cell., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Ray florets (Rf) and disc florets (Df) are agricultural byproducts of sunflower seeds. Their nutrition-related compounds were determined. The dietary fiber contents in Rf and Df were 42.90 mg/100 g and 58.97 mg/100 g. In both florets, palmitic, linoleic, and linolenic acids were identified as the three most abundant fatty acids, and the saturated ones constitute approximately two-thirds (w/w) of the total fatty acids. Lysine was the limiting amino acid in both florets by World Health Organization standards. Sixteen phenolic compounds, nine free and eight bound, mainly depsides, were identified in florets by RP-HPLC-DAD/ESI-TOF-MS. The free and bound phenolic compounds in Df were higher than in Rf. 1,5-di-O-caffeoylquinic acid was the predominant free phenolic compound in both florets. The present study revealed that the florets of sunflower are rich sources of dietary fiber, Fe, and phenols.

Objectives/hypothesis: Cell therapy has been shown to prevent vocal fold scarring and atrophy. However, problems that include the expansion of large numbers of cells in vitro and the poor survival of transplanted cells in vivo must be solved. The aim of this study was to use micronized acellular dermal matrix (MADM) as an expansion substrate of rabbit allogeneic adipose-derived stem cells (ADSCs) and to apply the combination of the matrix and cells, ADSC-MADM, to vocal fold regeneration., Study Design: Animal experiment., Methods: The proliferation of ADSCs that were cultured on the MADM was evaluated using 3-(4,5-dimethylthizazol-2-yl)-5-(3-carboxymethoxy-phenyl)-2-(4-sulfonyl)-2H-tetrazolium, and a rabbit acute vocal fold injury model was established by laser injury. Eighteen New Zealand white rabbits were randomly divided into three groups, which were injected with ADSC-MADM, ADSCs, and MADM, respectively. Morphological analysis was performed by laryngoscope, and histological analyses were indicated by hematoxylin and eosin staining, van Gieson staining, and immunofluorescence. Additionally, the in vivo survival of the ADSCs was determined by CM-Dil cell labeling., Results: When compared with a two-dimensional culture, the MADM significantly promoted proliferation of ADSCs. Morphological and histological analyses indicated that, when compared to only using of MAMD or the nontreatment sample, the use of ADSC-MADM or only using ADSCs successfully prevent scarring and atrophy. Moreover, ADSC-MADM exhibited a better therapeutic effect than when only using ADSCs, which was probably due to the MADM significantly enhancing the survival of transplanted ADSCs., Conclusions: MADM could be used as an efficient expansion substrate and delivery vehicle for ADSCs in vocal fold regeneration., (Copyright © 2012 The American Laryngological, Rhinological, and Otological Society, Inc.)

Light-harvesting complexes (LHCs) play essential roles in light capture and photoprotection. Although the functional diversity of individual LHCs in many plants has been well described, knowledge regarding the extent of this family in the majority of green algal groups is still limited. In this study, two different LhcSR genes, LhcSR1 and LhcSR2 from Chlamydomonas sp. ICE-L, were cloned from the total cDNA and characterized in response to high light (HL), low light (LL), UV-B radiation and high salinity. The lower F (v)/F (m) as well as the associated induction of non-photochemical quenching (NPQ), observed under those conditions, indicated that Chlamydomonas sp. ICE-L was under stress. Under HL stress, the expression of LhcSR1 and LhcSR2 increased rapidly from 0.5 h HL and reached a maximum after 3 h. In LL, LhcSR2 expression was up-regulated during the first 0.5 h after which it decreased, while the expression of LhcSR1 decreased gradually from the beginning of the experiment. In addition, the transcript levels of LhcSR1 and LhcSR2 increased under UV-B radiation and high salinity. These results showed that both genes were inducible and up-regulated under stress conditions. A higher NPQ was accompanied by the up-regulated LhcSR genes, suggesting that LhcSR plays a role in thermal energy dissipation. Overall, the results presented here suggest that LhcSR1 and LhcSR2 play a primary role in photoprotection in Chlamydomonas sp. ICE-L under stress conditions and provide an important basis for investigation of the adaptation mechanism of LhcSR in Antarctic green algae.

Few data are available about the effects of complexation of polyphenols with polysaccharide on their bioavailability. The complex of tea polyphenols (TP) with oat β-glucan was characterized by ultraviolet-visible spectrometry, Fourier transform infrared spectrometry, differential scanning calorimetry, atomic force microscopy, and solid-state (13)C NMR spectroscopy. The results indicated that the bonds which governed the interaction between TP and oat β-glucan were strong hydrogen bonds. The in vitro antioxidant activity of TP, β-glucan, their complex, and physical mixture was assessed using four systems, namely, DPPH(•), OH(•), and O(2)(•-) scavenging activities and reducing power. The complexation and blending of TP and β-glucan exhibited different impacts on the index of in vitro and in vivo antioxidant capacities. In the concentration range of 0.5-2.5 mg mL(-1), the complex had highest O(2)(•-) scavenging activity, whereas the highest OH(•) scavenging activity was found with the physical mixture. For antioxidant testing in vivo, there was no significant difference between the complex and the physical mixture in terms of glutathione peroxidase activity and levels of malondialdehyde and total antioxidant capacity in serums. However, the complex exhibited much higher activities of superoxide dismutase and glutathione peroxidase in livers than the physical mixture. The present study provided a deeper understanding of the influence of molecular interaction between TP and oat β-glucan on their antioxidant activities.

Objective: To investigate the age related changes of the expression of programmed cell death 5 (PDCD5) and caspase 3 in the cochlea of the different age of C57BL/6J mice. The relationship of PDCD5 and caspase 3 and the possible roles in the pathogenesis of presbycusis were also discussed., Methods: C57 mice of 3, 6, 9 and 12 months old were selected and divided into 4 groups, with 15 mice in each group. Auditory function of C57BL/6J mice was measured by auditory brainstem response (ABR) respectively. The changes of PDCD5 and Caspase 3 protein in the cochlea were detected by immunohistochemistry and Western blotting, the changes of PDCD5 mRNA and caspase 3 mRNA were detected using RT-PCR., Results: With the increase of age, the mean value for ABR thresholds in response to click, 4 kHz and 8 kHz sound stimulus of C57 mice gradually increased, the expression of PDCD5 and caspase 3 were increased also. At 3 months and 6 months of age in the cochlea of C57, all sorts of expression of PDCD5 and caspase 3 and the expression were enhanced with age. There was an evident expression at 9 months age, but the highest expression was detected at 12 months age. The PDCD5 and Caspase 3 expression were statistically different in each group (P < 0.05). The changes of PDCD5 and caspase 3 mRNA expression were in accordance with that of PDCD5 and Caspase 3 protein expression by the real-time PCR., Conclusions: The expression levels of PDCD5 and caspase 3 in the cochlea of C57 mice increase with age, the results suggested that the expression of PDCD5 and caspase 3 might play an important role in the pathogenic mechanism of presbycusis.

In addition to bone quantity, bone quality affects bone strength. Bone quality depends in part on the degree of mineralization of bone tissue (DMB). The relationship between the DMB distribution and the risk of osteoporotic hip fractures remains incompletely investigated. Here, our aim was to compare DMB distribution in femoral neck cortex specimens from 23 women with hip fractures (age, 65-96 years) and 14 control women (age, 75-103 years). Mineralization was determined using quantitative microradiography. We evaluated the following parameters of DMB frequency histograms, for both osteons and interstitial tissue: mode (oDMB(Al)mode and intDMB(Al)mode, respectively); 25th (oDMB(Al)q25, intDMB(Al)q25), 50th (oDMB(Al)q50, intDMB(Al)q50), and 75th (oDMB(Al)q75, intDMB(Al)q75) percentiles; and interquartile range (oDMB(Al)iqr, intDMB(Al)iqr). For each specimen, we also calculated the variance of pixel mineral content for osteons and interstitial tissue (oDMB(Al)var and intDMB(Al)var). We used nonparametric tests to compare frequency histogram parameters between hip-fractured women and controls and Fisher's test to compare variances between groups. All frequency histogram parameters for osteons and interstitial tissue except the 25th percentile, and the variances of pixel mineral content in osteons and interstitial tissue, were significantly different between hip-fractured women and controls, indicating greater heterogeneity of mineralization in the hip-fracture patients than in the controls. These cross-sectional data suggest that bone fragility may be related to greater DMB heterogeneity in osteons and interstitial tissue., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Objective: To investigate the presence of mesenchymal stem cells (MSC) in laryngeal mucosa, and to seek for the method to isolate, cultivate and identify these cells., Methods: Normal laryngeal mucosa was obtained from patients with laryngeal carcinoma during surgery, and the generating mesenchymal cells was obtained by digestive method. The cell growth curve was evaluated by 3-(4.5-methylthiazol-2yl)-2.5-diphenyltetrazolium bromide (MTT) assay. Colony forming cell assay was used to screen different morphologic colonies and evaluate clone formation ability. Flow cytometry was performed for the expression of the cells of the 4th passage surface marker profiles. Multiple differentiation potentials were confirmed by adipogenic, osteogenic and neural lineages induction., Results: MTT assay and colony forming cell assay showed that laryngeal mucosa MSC had a relatively rapid proliferation capacity and a relatively high clone formation capacity (clone formation rate 7.1%). Flow cytometry analysis revealed that the laryngeal mucosa MSC were positive for CD29 (16.9%), CD44 (97.4%), CD90 (89.5%), CD105 (85.9%), CD146 (2.5%) and stro-1 (20.4%), but negative for CD34 (1.2%) and CD45 (0.8%). Laryngeal mucosa MSC undergone adipogenic, osteogenic and neural lineages induction were positive for oil red staining, alizarin red staining and S100 staining respectively, which suggested that laryngeal mucosa MSC could differentiate into adipogenic, osteogenic and neural lineages., Conclusion: This study demonstrated that MSC with rapid proliferative capacity and multiple differentiation potential could be obtained from lamina propria of laryngeal mucosa.

Objective: To explore the feasibility of adipose-derived stem cells (ASC) combined with micronized acellular dermal matrix (MADM) for vocal cord injection., Methods: The adipose-deprived stem cells were harvested from rabbit adipose tissue in vitro. The 3rd generation of ASC was labeled with DiI (1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine perchlorate) and cultured with MADM to form a complex. The adhesion of ASC to MADM was observed by fluorescence microscope and electron microscope. The proliferation of ASC on MADM was evaluated by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy methoxyphenyl)-2-(4-sulfonyl)-2H-tetrazolium, inner salt (MTS). Three days after the culture, the complex was mixed with appropriate amount of collagen, and then injected into the unilateral vocal cord of the rabbit. The animals were sacrificed 2, 4, 8 weeks after injection, the survival time and distribution of ASC in vocal fold were tested, and the responses of vocal cord to ASC-MADM and the degradation of MADM were observed., Results: The ASC adhered to MADM and grew well (P < 0.05 or < 0.01), showing good compatibility with MADM in vocal cord tissue. The complex of ASC-MADM could be injected into the rabbit vocal cords, while no adverse reactions was observed in the vocal cord by endoscope, frozen section and HE staining. ASC could survive for 8 weeks in vocal cords, and no inflammatory cell infiltration was observed., Conclusions: MADM is an ideal scaffold material and shows perfect compatibility with ASC which can adhere and proliferate well on it. The complex of ASC-MADM can be injected into the vocal cord and can survive. There is no adverse reaction in vocal cords.

Oxidative stress plays an important role in Parkinson's disease and other neurodegenerative disorders. Salidroside, a phenylpropanoid glycoside isolated from Rhodiola rosea L., has potent antioxidant properties. In the present study, we investigated the protective activity of salidroside against 1-methyl-4-phenylpyridinium (MPP(+))-induced apoptosis in PC12 cells. We found that incubation of PC12 cells with salidroside prior to MPP(+) exposure significantly reduced cell apoptosis and attenuated collapse of the mitochondrial membrane potential (MMP). Furthermore, salidroside inhibited the MPP(+)-induced nitric oxide (NO) increase and overexpression of nNOS and iNOS and suppressed accumulation of reactive oxygen species (ROS) and intracellular free Ca(2+). Our results show that the protective effects of salidroside on PC12 cells are mediated, at least in part, by inhibition of the NO pathway., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Objective: To investigate the age-related expression of KCNQ1 and NKCC1 ion transporters in the stria vascularis in the cochlea of C57BL/6J mice, and to analyze the relationship between the two ion transporters and age-related hearing loss., Methods: Auditory function of C57BL/6J mice was measured by auditory brainstem response (ABR) at the ages of 4, 8, 14, 24, 40 weeks old respectively. The location of KCNQ1 and NKCC1 ion transporters in the cochlea of C57BL/6J mice was detected by immunohistochemistry staining. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the levels of KCNQ1 and NKCC1 mRNA in the cochlea of C57BL/6J mice at different ages., Results: The mean values for ABR thresholds in response to click, 4 kHz and 8 kHz sound stimulus of C57BL/6J mice gradually increased with age. The ABR thresholds of the mice of over 14 weeks age were significantly elevated in comparison with lower ages (P < 0.05). In the lateral wall of C57BL/6J mice cochlea, the KCNQ1 protein was mainly expressed at the apical membrane of the strial marginal cells. The localization of NKCC1 protein was mainly present at the basolateral membrane of the stria marginal cells, spiral ligament and the fibrocytes in the inferior portion of spiral limbus. Expression of KCNQ1 and NKCC1 protein in cochlea of C57BL/6J mice showed age-related decreasing. The level of KCNQ1 and NKCC1 mRNA in cochlea of C57BL/6J also showed a age-related decreasing trend. There was a significant reducing of KCNQ1 mRNA level between C57BL/6J mice of 40 and 4 weeks old (P < 0.05). In comparison with the C57BL/6J mice of 4 weeks old, the NKCC1 mRNA levels of 24 and 40 weeks old also showed significant reducing (P < 0.05)., Conclusions: The mean value for ABR thresholds of C57BL/6J mice gradually increased with age. Expression of KCNQ1 and NKCC1 protein in the stria vascularis of C57BL/6J mice decreases with age. The levels of KCNQ1 and NKCC1 mRNA in cochlea of C57BL/6J showed a age-related reducing trend. Regulating after post-translation may also participate in the adjusting of the age-related decreasing of KCNQ1 and NKCC1 protein in the cochlea of C57BL/6J mice. KCNQ1 and NKCC1 ion transporters may play a critical role in maintaining normal hearing function of inner ear.

Laser tweezers Raman spectroscopy can help with observing and studying individual cells or organelles in a natural state for a relatively long period. In the present experiment, Laser tweezers Raman spectroscopy (LTRS) was used as a tool to report physiological metabolism such as cells growth and nucleic acid, proteins, lipid and glucose of a single active cold-adapted Aromatic hydrocarbons-degradating strains isolated from Antarctic Sea. After the Raman spectrum was collected and analyzed, the findings are as follows: Raman spectrum identified the components of a single cold-adapted Aromatic hydrocarbons-degradating strain and there were more proteins and carbohydrate produced during the Planococcus sp. NJ41 and Shewanella sp. NJ49 growth and degradation; but there was more lipid than the proteins produced during the Pseudoalteromonas sp. NJ289 growth and degradation; the amount of proteins produced by the strains corresponds with the production of degradation rate-limiting enzyme, and was also related to the capacity of low-temperature degradation of aromatic hydrocarbons.

The title anti-pyrine derivative, C(22)H(19)N(3)O(2), was synthesized by the reaction of 4-amino-1,5-dimethyl-2-phenyl-1,2-dihydro-pyrazol-3-one and 1-hy-droxy-naphthalene-2-carbaldehyde in methanol solution. As expected, the compound adopts a trans configuration about the central C=N bond. The N atom is involved in an intra-molecular O-H⋯N bond which stabilizes the mol-ecular configuration. In the crystal structure, adjacent mol-ecules stack with no short contacts.

Objective: To introduce a novel retractor with magnetic fixator for mouse microsurgery., Methods: The retractor was consisted of a magnet, a screw, two screw nuts and a hook made of dental stainless wire. The screw was connected to the magnet with magnetic force, and then was assembled to be a so-called magnetic fixator. The hook was clamped by two screw nuts on the screw, and these makes up of the retractor finally. Comparison has been done between the novel retractor and traditional retractor in the clinical application of the otocyst exposure., Results: The retractor can quickly claw and retract massive tissue like muscles and vessels to the target position, thus, this properties would tend to offer a clear and expanded operative field. In addition, the height, orientation and strength of traction was all adjustable. By Comparison with tradition retractor, the operative incision can be shorten via the application of the retractor, also, it would reduce the trauma of muscles and vessels as well as the accidental rate of bleeding in the process of operation., Conclusions: The retractor can offer a expanded operative field of the mouse otocyst conveniently. It could be a simple, powerful and minimal invasive tool for mouse microsurgery.

Objective: To investigate the ototoxicity of co-administration of kanamycin and furosemide in mouse and establish a reliable model to induce a sensorineural hearing loss., Methods: CBA/J mice strain was selected, with the age around 3-4 weeks old, to be received a single subcutaneous injection of kanamycin at dose of 1 g/kg and another single intraperitoneal injection of furosemide at dose of 0.4 g/kg 30 - 45 min afterward. The auditory brainstem response (ABR) threshold shift was tested. The series of experimental methods including propidium iodide, phalloidin staining, semithin section toluidine blue staining, TUNEL, scanning electron microscopy and transmission electron microscopy were applied to observe the characteristics of the lesion of cochlea and hair cells. The time course was set as following: before injection, 12, 24, 48 hours and 1, 2, 4, 12 weeks after injections, respectively., Results: The ABR threshold shift was firstly presented a significant increase at 12 h after injection at 2, 4, 8 kHz, then the ABR threshold kept going up during next 36 h until it was presented a stable level around 90 dB. Pathological examination showed an absence of outer hair cells at basal turn rapidly since 12 h after treatment, and then by 48 h the most commonly observed lesion, where all outer hair cells throughout the length of the cochlea were killed, in the contrast, however, the inner hair cells loss were delayed and mild. TUNEL-positive nuclei demonstrated that most hair cells died via an apoptotic pathway. In scanning electron microscopy abundance of necrotic outer hair cells were detected by 24 h after treatment, in which reticular lamina were collapsed. Then all outer hair cells were replaced by expansion of heads of supporting cells. At 48 h after treatment, marginal cells presented a swollen and some of them were observed to be fused. In addition, spherical cell extrusion appeared to leak out from some marginal cells. By 2 weeks, nearly all microvillus were lost and marginal cells presented a shape of stone-like change. A significant and progressive decrease in strial vascularis thickness was found, of which the reason probably related with a reduction in volume of marginal cells., Conclusion: This systemic protocol eliminates hair cells extensively in vivo, and it could be a reliable model to examine different aspects of cochlear pathology in transgenic or mutant mice strains.

The title compound, [C(8)H(18)N](+)·[B(5)O(6)(OH)(4)](-), has been synthesized under mild solvothermal conditions in the presence of N,N-dimethyl-cyclo-hexyl-amine acting as a template. The structure consists of penta-borate [B(5)O(6)(OH)(4)](-) anions connected through O-H⋯O hydrogen bonds into a three-dimensional framework, with large channels along [100], [010] and [001] directions. The [C(8)H(18)N](+) cations reside in the channels, inter-acting with the framework through N-H⋯O hydrogen bonds.

Objective: To investigate the auditory function and the role of NKCC1 and alpha2 Na, K-ATPase in the potassium recycling of cochlea., Methods: NKCC1(+/-) / alpha2 Na, K-ATPase(+/-) mice model was established from NKCC1(+/-) and alpha2 Na, K-ATPase(+/-) mice. The auditory function of all strain mice were detected by auditory brainstem response (ABR) and endocochlear potential (EP) to investigate the role of NKCC1 and alpha2 Na, K-ATPase in the potassium recycling of cochlea. Furosemide and ouabain were applied to block the two channels in Castel mice line which can long-time maintain normal auditory function and then their auditory function was detected by ABR to authenticate the mode of potassium recycling in vivo and the relationship between cochlear potassium recycling and NKCC1(+/-) and alpha2 Na, K-ATPase., Results: The mean value for ABR thresholds in response to stimulus was elevated in NKCC1(+/-) and alpha2 Na, K-ATPase (+/-) mice [(38.49 +/- 12.29) dB and (53.32 +/- 7.62) dB) ] respectively, which was significantly increased compared with that observed in wild type mice [(23.13 +/- 3.78) dB, P < 0.05) ]; The EP value of NKCC1(+/-) [(78 +/- 7) mV] and alpha2 Na, K-ATPase(+/-) mice [(71 +/- 14) mV] was decreased compared with that of NKCC1(+/-) / alpha2 Na, K-ATPase(+/-) mice [( 86 +/- 11) mV]. The auditory function of NKCC1(+/-) / alpha2 Na, K-ATPase(+/-) mice could simulate the model of cochlear potassium recycling well. NKCC1 and Na, K-ATPase were great of importance in the potassium recycling, while the two ion channels were in restrict dynamic equilibrium. Castel mice line after administration with furosemide developed significant ABR threshold shifts (P < 0.05) compared with control group. Castel mice line after administration with ouabain also developed greatly significant ABR threshold shifts (P < 0.05) compared with control group. ABR threshold shifts in mice after administration both furosemide and ouabain was attenuated compared with only administration with furosemide (P < 0.01)., Conclusions: Ion channel NKCC1 and alpha2 Na, K-ATPase played important roles in the inner ear potassium recycling. Dysfunction of either of them could influence potassium concentration in the endolymph and lead to hearing loss subsequently. The role of NKCC1 and alpha2 Na, K-ATPase in cochlear potassium recycling was authenticated in vivo. The two ion channels contribute the key role for dynamic equilibrium in cochlear potassium recycling and are of great importance for the metabolism of potassium in the inner ear to maintain the normal auditory function.

Grain size and shape are important factors affecting grain quality and yield in rice. Mapping, tagging and identification of their related genes can lead us to understand their expression pattern and mechanism network, which is to their control. In this study we mapped a grain length controlling gene named Lk-4(t) with SSR and CAPs markers by screening 800 recessive plants in a BC2F2 population derived from a cross of Shuhui527xXiaoli and backcrossed with Xiaoli as the donor parent. The distribution of grain shape parameters and thousand grain weight in F2 and BC2F2 population showed that backcross can diminish most unnecessary variations to identify the target gene more clearly. There were only two grain length phenotypes found among the 3 209 BC2F2 plants, long and short, indicating it is a qualitative trait. The frequency distribution for the grain length showed a typical segregation ratio of 3:1, suggesting that only one allele was responsible for the variation. By screening the recessive long grain plants with three CAPs markers, P1-EcoRV, P2-Sac I and P3-Mbo I, we tagged the locus on the arm of chromosome 3 near the centromere. Lk-4(t) was located between P1- EcoRV and P2-Sac I, with genetic distance of 0.90 cM and 0.50 cM from the two markers respectively. Mapping of the gene is a foundation for its final identification and function analysis.

Background and Purpose: Previously reported quantitative parameters for the magnetization transfer ratio (MTR) do not give identical results, which can limit their ability to differentiate normal from diseased tissue and render them vulnerable to variations among MR systems. Our purpose was to systematically study different MTR metrics; propose a new MTR histogram parameter, AMTR(2/3); and compare AMTR(2/3) with existing parameters in a study of multiple sclerosis (MS)., Methods: Seven conventional MTR parameters were proposed: global and mean MTR; peak height and position of the histogram; and percentiles MTR25, MTR50, and MTR75. Additionally, we investigated a parameter, AMTR(2/3), to indicate the normalized pixel count (area under the histogram curve) inside the band size of two-thirds MTR histogram peak height. All parameters were measured in 10 patients with relapsing-remitting MS (group A), 10 healthy control subjects from the same imaging center as that of patients (group B), and four healthy control subjects from an outside institution (group C). Comparison of findings was performed between groups A and B to assess the discriminating ability of MTR parameters and groups B and C to evaluate intersystem variations., Results: All MTR parameters differed between groups A and B, but the difference was significant for only global MTR, mean MTR, MTR25, and AMTR(2/3). With the exception of AMTR(2/3), all parameters differed significantly between the two control groups., Conclusion: AMTR(2/3) is less sensitive to MR imaging system variations than are other MTR parameters and was most effective in differentiating patients with MS from healthy control subjects. This finding supports the use of AMTR(2/3) in multicenter MT MR imaging studies of MS.

The title compound, C(17)H(14)F(6)O(2)S(2), a photochromic diarylethene, is one of the most promising materials for optical memories and other optoelectronic devices. The hexafluorocyclopentene group and the two thiophene rings are all planar, and the dihedral angles between the cyclopentene ring and the adjacent thiophene rings are 46.4 (1) and 49.5 (1) degrees.

Objective: To crosslink bovine pericardial valves by phthalocyanine sensitized photooxidation reaction., Methods: Fresh bovine pericardial tissues were immersed in PBS solution containing 0.01% cupreous sulfonatophthalocyanine and exposed to a broad wavelength light source for 20-120 h. Using sodium dodecyl sulfate-polyarylamide gel electrophoresis (SDS-PAGE) to analyze the cross-linking extent of pericardial tissue. With the help of attenuated total reflection-fourier transform infrared (ATR-FTIR), explained the change of protein structure after photooxidation reaction. Differential scanning calorimetry (DSC) was used to measure the change of shrinkage temperature. Tissue water content experiment method was also used., Results: Photooxidized tissue was resistant to extraction when it was compared with untreated tissue and had similar crosslinking extent over glutaraldehyde-treated tissue. The three dimension structure of collagen were changed but not seriously destroyed and the model of photooxidation reaction was validated. Phthalocyanine-treated tissue was much more flexible than glutaraldehyde-treated tissue., Conclusion: Bovine pericardial tissue can be successfully crosslinked by phthalocyanine-mediated photooxidation reaction.