To investigate the evolutionary process by which porcine epidemic diarrhea virus (PEDV) in the United States hypothetically descended from strains in China, we analyzed PEDV-positive samples collected in China during January 2012–July 2013. Recombination in 2 strain sublineages was likely associated with identification of PEDV in the United States in 2013.
The autologous tumor cell vaccine and NDV vaccine can prolong the patients' life. NDV vaccine is notably effective for short-term with promotion of quality of life and can be used whenever necessary with good prospects.
Summary• The aim of this study was to investigate the physiological significance of increased proline loading to phloem caused by water-deficit stress in relation to nitrogen (N) uptake and assimilation.• N uptake and N assimilation were quantified by 15 N tracing in well-watered (control) and water deficit-stressed white clover (Trifolium repens). De novo proline synthesis and proline loading to the phloem were also compared between treatments. The relationships among proline concentrations in phloem exudates, N uptake, and assimilation of newly absorbed N were assessed.• The newly synthesized proline in the phloem exudates increased rapidly after 3 d of water deficit. The water-deficit treatment significantly reduced the maximum nitrate reductase activity (NRA), and also attenuated de novo synthesis of amino acids and proteins in the roots. The increase in proline concentrations in phloem exudates was closely related to reductions in NRA in the roots, N uptake, and the assimilation of newly absorbed N. The accumulation of proline induced in roots by exogenous proline and NH 4 Cl treatments was closely associated with the decrease in NRA.• These results indicate that increased proline transport to roots via phloem caused by water deficit has a significant influence on the down-regulation of N uptake and the assimilation of newly absorbed N.
Suppression of ␣-mannosidases by chemicals has been shown to reduce the potentiality of growth and metastasis of various tumors. In our study, the effect of 6A8 ␣-mannosidase (MAN 6A8), recently discovered in our laboratory, on malignant behaviors of tumor cells was examined. Since the suppressive effect of chemicals on ␣-mannosidase is not specific, antisense technique was used to specifically inhibit expression of the MAN 6A8 in human nasopharyngeal carcinoma cells, CNE-2L2.
Key words: ␣-mannosidase; tumor; growth; metastasis; antisense; E-cadherinInhibition of ␣-mannosidase has been found to show suppressive effect on malignant behaviors of tumor cells.  In examining the effect of ␣-mannosidase on growth and metastasis of tumors, chemicals, such as swainsonine, 1-deoxymannojirimycin, mannostatin, etc., have been normally used to inhibit ␣-mannosidase. 1,3-5 Among the chemicals, swainsonine is the one rather widely used that has been tested even in preliminary clinical trials in late-stage cancer patients and was shown to induce partial remission of tumor in 1 and symptomatic improvement in 2 of 19 patients. 6,7 Recently, we isolated a cDNA encoding a novel human ␣-mannosidase (MAN 6A8). 8 In searching the EST database of the Cancer Genome Anatomy Project for 6A8 gene expression in various tissues, we found an enhancement of 6A8 gene expression in a number of cancer tissues, including bone, bone marrow, brain, esophagus, eye, genitourinary, germ cell, nervous, pineal gland, soft tissue cancers, etc., suggesting a possible role of MAN 6A8 in tumorigenesis. Hence, the relationship between MAN 6A8 gene expression and malignant behaviors of tumor cells has been critically examined. Our study was carried out on a human nasopharyngeal carcinoma cell line, CNE-2L2. Because the chemicals mentioned above cannot specifically suppress a given ␣-mannosidase and usually have side effects 1 and particularly because swainsonine has no effect on MAN 6A8, 8 we used the antisense technique 9 to inhibit mRNA translation of MAN 6A8. It was found that the malignant behaviors of CNE-2L2 cells, both in vitro and in vivo, were significantly reduced when MAN 6A8 expression was inhibited. Since morphologic observation showed that cultured CNE-2L2 cells with inhibited MAN 6A8 expression became compact sheets instead of being dispersed as compared to the controls and since cell-cell adhesion of epithelial cells is mainly mediated by E-cadherin 10 and loss of E-cadherin expression plays an important role in the growth and metastasis of tumors, 11-16 expression of E-cadherin was assayed in CNE-2L2 cells. An enhancement of E-cadherin expression was found in cells with inhibited MAN 6A8 expression.The results demonstrate a significant inhibitive effect of reduced expression of MAN6A8 on the malignant behaviors of CNE-2L2 cells and suggest the possibility of the enzyme being a target molecule in the treatment of nasopharyngeal carcinoma.
MATERIAL AND METHODS
Cell line and animalCNE-2L2 is a cloned cell line isolated from CNE-2Z, a h...
BackgroundPrimary spontaneous pneumothorax (PSP) or pulmonary cysts is one of the manifestations of Birt-Hogg-Dube syndrome (BHDS) that is caused by heterozygous mutations in FLCN gene. Most of the mutations are SNVs and small indels, and there are also approximately 10 % large intragenic deletions and duplications of the mutations. These molecular findings are generally obtained by disparate methods including Sanger sequencing and Multiple Ligation-dependent Probe Amplification in the clinical laboratory. In addition, as a genetically heterogeneous disorder, PSP may be caused by mutations in multiple genes include FBN1, COL3A1, CBS, SERPINA1 and TSC1/TSC2 genes. For differential diagnosis, these genes should also be screened which makes the diagnostic procedure more time-consuming and labor-intensive.MethodsForty PSP patients were divided into 2 groups. Nineteen patients with different pathogenic mutations of FLCN previously identified by conventional Sanger sequencing and MLPA were included in test group, 21 random PSP patients without any genetic screening were included in blinded sample group. 7 PSP genes including FLCN, FBN1, COL3A1, CBS, SERPINA1 and TSC1/TSC2 were designed and enriched by Haloplex system, sequenced on a Miseq platform and analyzed in the 40 patients to evaluate the performance of the targeted-NGS method.ResultsWe demonstrated that the full spectrum of genes associated with pneumothorax including FLCN gene mutations can be identified simultaneously in multiplexed sequence data. Noteworthy, by our in-house copy number analysis of the sequence data, we could not only detect intragenic deletions, but also determine approximate deletion junctions simultaneously.ConclusionsNGS based Haloplex target enrichment technology is proved to be a rapid and cost-effective screening strategy for the comprehensive molecular diagnosis of BHDS in PSP patients, as it can replace Sanger sequencing and MLPA by simultaneously detecting exonic and intronic SNVs, small indels, large intragenic deletions and determining deletion junctions in PSP-related genes.
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