[Optimal time window for observation of calcific aortic valve disease in mice following catheter-induced valve injury].

OBJECTIVE: To investigate the optimal time window for observation of catheter-induced valve injury that mimics calcified aortic valve disease in mice. METHODS: A catheter was inserted into the right common carotid artery of 8-week-old C57BL6 mice under ultrasound guidance, and aortic valve injury was induced using the guide wire.At 4, 8 and 16 weeks after modeling, the mice were subjected to ultrasound measurement of the heart short axial shortening rate, aortic valve peak velocity and aortic valve orifice area.Grain-Eosin staining was used to observe the changes in the thickness of the aortic valve, and calcium deposition in the aortic valve was assessed using Alizarin red staining.Immunofluorescence assay was performed to detect the expression of alkaline phosphatase (ALP) in the aortic valve. RESULTS: At 4, 8 and 16 weeks after modeling, valve thickness (P=0.002), calcium deposition (P < 0.0001) and the expression of osteogenic protein ALP (P=0.0016) were significantly increased, but their increments were comparable at the 3 time points of observation. CONCLUSION: In mouse models of calcific aortic valve disease induced by catheter valve injury, 4 weeks after the injury appears to be the optimal time window for observation of pathophysiological changes in the aortic valves to avoid further increase of the death rate of the mice over time.

[Research on ZNA probe in the quantitative detection of chlamydia trachomatis nucleic acid].

Objective: To investigate the performance of ZNA(ZIP Nucleic Acid) probes and its application in the quantitative detection of Chlamydia trachomatis (CT)nucleic acid. Methods: Use CT positive plasmids to compare the PCR amplification curves of ZNA probes coupled with different ZIP numbers. Compare ZNA probes with other three sets of probes [namely, 29mer ordinary Taqman probes (long-DNA probe), 20mer ordinary Taqman probes (short-DNA probe) and MGB probes] for stability in PCR amplification curves and repeated freezing and thawing, and the difference in the detection rate of low-concentration plasmids. Use CT positive clinical samples to compare the difference in amplification curves between ZNA probes, long-DNA probe, short-DNA probe and MGB probes, and the detection rate of low-concentration samples. Results: (1) The Ct value and fluorescence value of the probes coupled with 5ZIP units are both better than those coupled with a smaller number of ZIPs. And the difference is biggest when compared with only coupled with 1 ZIP unit: Ct value increased by 1.34 (sensitivity increased by 2.37 times), and fluorescence value increased by 30%. (2) The amplification efficiency of the ZNA probe coupled with 5 ZIPs is 2.14-2.64 times that of the preferred ordinary Taqman probe and MGB probe, and the fluorescence value is 17%-90% higher. (3) The probe freeze-thaw stability results show that the ZNA probe has the best stability, and the lowest concentration of Ct value has the smallest deviation (CV% = 1.4), which is better than the other three sets of probes (CV%=1.7-3.7). (4) Using 35 CT positive clinical samples to compare the PCR amplification performance, compared with other three sets of probes, the amplification sensitivity of ZNA probes was increased by 1.60, 0.99 and 1.06 times respectively. And the results of the consistency analysis of the Ct value show that compared with short-DNA probe and MGB probes, ZNA probes have better detection performance for clinical samples. (5) Use low concentration plasmid template (200, 100, 50 and 10 copies/mL respectively) to compare the amplification sensitivity of the four sets of probes, the detection rate of ZNA probe is the best. Especially, at the lowest concentration 10 copies/mL, the detection rate of the other sets of probes is only 15%-20%, but the ZNA probe is still 30%. (6) In 20 clinical samples with different low concentrations (200, 150, 100, and 50 copies/mL), the detection rate of ZNA probes was the highest, which were 100%, 95%, 90%, and 70%, respectively. Conclusions: Through testing of the amplification efficiency, fluorescence value, freeze-thaw stability, the amplification performance of clinical samples and the detection sensitivity of low-concentration samples, ZNA probes coupled with 5 ZIPs show better performance than ordinary Taqman probes and MGB probes. As a new probe technology with flexible design and easy synthesis, ZNA probe can further improve detection sensitivity of low concentration samples in the field of gene expression.

Accuracy assessment of NLCD 2011 impervious cover data for the Chesapeake Bay region, USA.

The National Land Cover Database (NLCD) contains three eras (2001, 2006, 2011) of percentage urban impervious cover (%IC) at the native pixel size (30 m-×-30 m) of the Landsat Thematic Mapper satellite. These data are potentially valuable to environmental managers and stakeholders because of the utility of %IC as an indicator of watershed and aquatic condition, but lack an accuracy assessment because of the absence of suitable reference data. Recently developed 1 m2 land cover data for the Chesapeake Bay region makes it possible to assess NLCD %IC accuracy for a 262,000 km2 region based on a census rather than a sample of reference data. We report agreement between the two %IC datasets for watersheds and the riparian zones within watersheds and four additional square units. The areas of the six assessment units were 40 ha cell, 433 ha (riparian mean), 2756 ha cell, 5626 ha cell, 8569 ha (watershed mean) and 22,500 ha cell. Mean Absolute Deviation (MAD) and Mean Deviation (MD) were about 1.5% and -1.5%, respectively, for each of the assessment units except for the riparian unit, for which MAD and MD were 0.88 and 0.62, respectively. NLCD reliably reproduced %IC from the 1 m2 data with a small, consistent tendency for underestimation. Results were sensitive to assessment unit choice. The results for the four largest assessment units had very similar regression parameters, R2 values, and bias patterns. Results for the riparian assessment were different from those for the watershed unit and the other three larger units. MAD was about 50% less for the riparian zones than it was for the watersheds, the direction of bias was less consistent, and NLCD %IC was uniformly higher than 1 m2 %IC in urbanized riparian zones. For the smallest unit, bias patterns were more similar to the riparian unit and regression results were more similar to the four larger units. MAD and MD were also sensitive to the amount of urbanization, increasing as NLCD %IC increased. The low overall bias and positive relationship between bias and urbanization suggest that the benefits of obtaining 1 m2 IC data outside of urban areas may not outweigh the costs of obtaining such data.

New Record of the Scrub Typhus Vector, Leptotrombidium rubellum, in Southwest China.

There are several main vectors of scrub typhus in China, and Leptotrombidium rubellum Wang et Liao, 1984 is one of them, which was previously considered to be restricted to the coastal regions of Changle to Xiamen, Fujian province of east China. Ecological investigation of chigger mites in recent years demonstrated the presence of L. rubellum also in Yunnan province. Chigger mites were collected from 34 counties, in which 127,460 larval mites were collected from 14,381 small mammal hosts. A total of 277 species belonging to 26 genera and three subfamilies were identified. A total of 705/127,460 (0.55%) L. rubellum were collected from eight counties. Leptotrombidium rubellum was collected mainly at low elevations in southern Yunnan. A total of 663/705 (94.04%) of L. rubellum were collected from Rattus flavipectus (Milne-Edwards, 1871) found in outdoor habitats with relatively high infestation prevalence and mean intensity. Although it was collected from several hosts, the primary host was Rattus tanezumi. This represents a new distribution record of L. rubellum for Yunnan province.

Cholesterol oxidase from Bordetella species promotes irreversible cell apoptosis in lung adenocarcinoma by cholesterol oxidation.

Cholesterol oxidase (COD), an enzyme catalyzing the oxidation of cholesterol, has been applied to track the distribution of membrane cholesterol. Little investigations about the effect of COD on tumor cells have been performed. In the present study, we provided evidence that COD from Bordetella species (COD-B), induced apoptosis of lung cancer cells in vitro and in vivo. COD-B treatment inhibited Akt and ERK1/2 phosphorylation in dose- and time-dependent manner, which was not reversed and was even aggravated by cholesterol addition. Further investigation indicated that COD-B treatment promoted the generation of reactive oxygen species (ROS) and that cholesterol addition further elevated ROS levels. Moreover, COD-B treatment resulted in JNK and p38 phosphorylation, downregulation of Bcl-2, upregulation of Bax, activated caspase-3 and cytochrome C release, which likely responded to freshly produced hydrogen peroxide that accompanied cholesterol oxidation. Catalase pretreatment could only partially prevent COD-B-induced events, suggesting that catalase inhibited H2O2-induced signal transduction but had little effect on signal pathways involved in cholesterol depletion. Our results demonstrated that COD-B led to irreversible cell apoptosis by decreasing cholesterol content and increasing ROS level. In addition, COD-B may be a promising candidate for a novel anti-tumor therapy.

CHD1L is a novel independent prognostic factor for gastric cancer

BACKGROUND: Chromodomain helicase/ATPase DNA binding protein 1-like gene (CHD1L) is involved in malignancies. However, the role of CHD1L in gastric cancer (GC) has not been elucidated. The aim of this study is to explore the clinical role of CHD1L in GC. METHODS: The gene and protein expression levels of CHD1L were detected by quantitative real-time PCR and Western blot analysis in fresh samples of GC and paired adjacent noncancerous tissue (n = 34). We evaluated the CHD1L expression by immunohistochemistry in a large number of GC patients (n = 616) and paired adjacent noncancerous tissues from December 1, 2004 to December 1, 2008. The correlations of CHD1L expression with clinicopathological features and clinical outcome were analyzed. RESULTS: The gene and protein expression levels of CHD1L were higher in fresh samples of GC than in paired adjacent noncancerous tissues as determined by quantitative real-time PCR and Western blot analysis. Immunohistochemical analysis showed that positive expression rates of CHD1L in GC and paired adjacent noncancerous tissues were 58.7 % (361/616) and 7.3 % (45/616), respectively. CHD1L positivity was significantly associated with clinical stage and distant metastasis. GC patients with positive CHD1L expression had shorter overall survival than those with negative CHD1L expression. Multivariate analysis showed that CHD1L was an independent prognostic marker for overall survival [Hazard Ratio (HR) = 5.952, 95 % confidence interval (CI) = 3.194-11.187, P = 0.0043]. CONCLUSION: These results indicated that CHD1L could serve as a prognostic marker for GC (AU)

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CHD1L is a novel independent prognostic factor for gastric cancer.

BACKGROUND: Chromodomain helicase/ATPase DNA binding protein 1-like gene (CHD1L) is involved in malignancies. However, the role of CHD1L in gastric cancer (GC) has not been elucidated. The aim of this study is to explore the clinical role of CHD1L in GC. METHODS: The gene and protein expression levels of CHD1L were detected by quantitative real-time PCR and Western blot analysis in fresh samples of GC and paired adjacent noncancerous tissue (n = 34). We evaluated the CHD1L expression by immunohistochemistry in a large number of GC patients (n = 616) and paired adjacent noncancerous tissues from December 1, 2004 to December 1, 2008. The correlations of CHD1L expression with clinicopathological features and clinical outcome were analyzed. RESULTS: The gene and protein expression levels of CHD1L were higher in fresh samples of GC than in paired adjacent noncancerous tissues as determined by quantitative real-time PCR and Western blot analysis. Immunohistochemical analysis showed that positive expression rates of CHD1L in GC and paired adjacent noncancerous tissues were 58.7 % (361/616) and 7.3 % (45/616), respectively. CHD1L positivity was significantly associated with clinical stage and distant metastasis. GC patients with positive CHD1L expression had shorter overall survival than those with negative CHD1L expression. Multivariate analysis showed that CHD1L was an independent prognostic marker for overall survival [Hazard Ratio (HR) = 5.952, 95 % confidence interval (CI) = 3.194-11.187, P = 0.0043]. CONCLUSION: These results indicated that CHD1L could serve as a prognostic marker for GC.

Plant Anticancer Agents XXXIII. Constituents of Passerina vulgaris1.

Two lignans of known structure, (+)-syringaresinol, a cytotoxic agent, and (+)-nortrachelogenin, a compound with demonstrated antileukemic activity, were isolated from a biologically active extract of the stems of PASSERINA VULGARIS.