Molecular identification and phylogenetic analysis of fungal pathogens isolated from diseased fish in Xinjiang, China.

The outbreaks of fungal diseases in cultured fish have been severe in recent years, which is harmful to the healthy and sustainable development of fish farming. In this study, an investigation was conducted for significant fungal infections of 12 species of fish in four regions in Xinjiang, China, to understand the distribution of local fish fungal pathogens. Twenty-six fungal strains with pathogenicity were isolated, and the challenge experiment showed that eight strains from Changji area had high infection rate to fish eggs. Based on internal transcribed spacer sequence data and molecular analysis, the 26 strains were classified into nine different species of six fungal genera. Phylogenetic analysis showed that all strains were divided into two clades, namely Cluster 1 (contains only the genus Mucor) and Cluster 2 (consists of five small branches), and the distribution of strains from the same region was scattered in two clusters. There is no strict host selectivity for these fungi to infect fish. Mucor sp. are the main fungal pathogen of fish in these four regions, whereas Hypophthalmichthys molitrix and Carassius auratus are two types of fish that were susceptible to pathogen. In addition, the environmental adaptability experiments showed that eight highly pathogenic strains have different adaptability to the environment, and their optimum temperature and pH were 25°C and 7.0, respectively, whereas the concentration of NaCl was negatively correlated with the growth of strains. Therefore, these results indicated that the coinfection of multiple fungal pathogens in a culture region should be considered in the future study.

A Specific High Toxicity of Xinjunan (Dioctyldiethylenetriamine) to Xanthomonas by Affecting the Iron Metabolism.

Xanthomonas spp. encompass a wide range of phytopathogens that brings great economic losses to various crops. Rational use of pesticides is one of the effective means to control the diseases. Xinjunan (Dioctyldiethylenetriamine) is structurally unrelated to traditional bactericides, and is used to control fungal, bacterial, and viral diseases with their unknown mode of actions. Here, we found that Xinjunan had a specific high toxicity toward Xanthomonas spp., especially to the Xanthomonas oryzae pv. oryzae (Xoo), the causal agent of rice bacterial leaf blight. Transmission electron microscope (TEM) confirmed its bactericidal effect by morphological changes, including cytoplasmic vacuolation and cell wall degradation. DNA synthesis was significantly inhibited, and the inhibitory effect enhanced with the increase of the chemical concentration. However, the synthesis of protein and EPS was not affected. RNA-seq revealed differentially expressed genes (DEGs) particularly enriched in iron uptake, which was subsequently confirmed by siderophore detection, intracellular Fe content and iron-uptake related genes transcriptional level. The laser confocal scanning microscopy and growth curve monitoring of the cell viability in response to different Fe condition proved that the Xinjunan activity relied on the addition of iron. Taken together, we speculated that Xinjunan exerted bactericidal effect by affecting cellular iron metabolism as a novel mode of action. IMPORTANCE Sustainable chemical control for rice bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae need to be developed due to limited bactericides with high efficiency, low cost, and low toxicity in China. This present study verified a broad-spectrum fungicide named Xinjunan possessing a specific high toxicity to Xanthomonas pathogens, which were further confirmed by affecting the cellular iron metabolism of Xoo as a novel mode of action. These findings will contribute to the application of the compound in the field control of Xanthomonas spp.-caused diseases, and be directive for future development of novel specific drugs for the control of severe bacterial diseases based on this novel mode of action.

[DNA methylation levels of genome in cFb transdifferentiation induced by SiO2 in rats].

OBJECTIVE: To explore the DNA methylation levels of genome in cFb transdifferentiation induced by SiO2 in rats. METHODS: The primary macrophages and fibrocytes of SD rats were co-cultured directly and indirectly, which were exposed to SiO2 at the doses of 25, 50 and 100 g/ml. The transdifferentiation of cFb was identified with immunohistochemical assay. The genomic DNA methylation levels of cFb were detected with HPLC. RESULTS: Under the condition of indirect co-culture, as compared with control group, the genomic DNA methylation levels of cFb exposed to SiO2 at the doses of 25, 50 and 100 g/ml reduced by 19.9%, 26.9% and 30.3%, respectively (P < 0.05); as compared with cFb exposed to 100 g/ml SiO2, the genomic DNA methylation levels of cFb exposed to 5-aza-dC decreased by 22.0% (P < 0.05). Under the condition of ThinCert(TM) direct co-culture, as compared with control group, the genomic DNA methylation levels of cFb exposed to SiO2 at the doses of 25, 50 and 100 g/ml reduced by 22.2%, 30.2% and 36.7%, respectively (P < 0.05); as compared with cFb exposed to 100 g/ml SiO2, the genomic DNA methylation levels of cFb exposed to 5-aza-dC decreased by 20.6% (P < 0.05). CONCLUSION: Under the co-culture condition in vitro, SiO2 could reduce the genomic DNA methylation levels of cFb. The ThinCert(TM) direct co-culture can be used to study the silicosis fibrosis.

Analysis of Ultrasonographic Characteristics of Early Hepatic Alveolar Echinococcosis.

Objective: This study aims to investigate the ultrasonographic characteristics of early hepatic alveolar echinococcosis (HAE) and improve the qualitative diagnostic ability of sonographers. Methods: The data of 80 positive cases of HAE screened and diagnosed by ultrasonography and serum immunology (33 males and 44 females with a mean age of 44.12 ± 14.31 y) were used in the study. The imaging characteristics of the lesions (i.e., intrahepatic distribution, shape, size, echo, blood flow, and growth characteristics) were analyzed retrospectively, and the ultrasonographic characteristics of early lesions were discussed in combination with their basic pathological changes. Results: Patients with lesions of the proliferative infiltration type accounted for 57.5% (46/80), the fibrous calcification type accounted for 26.25% (21/80), the necrotic liquefaction type accounted for 6.25% (5/80), and the mixed type accounted for 10% (8/80). Patients with lesions involving the right lobe and the left lobe accounted for 76.25% (61/80) and 11.25% (9/80), respectively, and with lesions involving both the left and right lobes accounted for 12.5% (10/80). There were statistically significant differences in diameter between all types of lesions (n = 88; F = 5.926 and P = 0.004). Focal hyperechoic and diffuse heterogenous nodular changes were the main manifestations of early proliferative infiltration lesions. Conclusion: Ultrasonography is extremely valuable in the diagnosis of early HAE. Capillary hemangioma-like changes and hailstorm sign are the main characteristics of early lesions of HAE, and calcification is a common concomitant sign.

[A meta-analysis of susceptibility to pneumoconiosis and polymorphism of tumor necrosis factor-α 308 and 238 locus].

OBJECTIVE: To explore the relationship between the polymorphism of TNF-α gene 308, 238 locus and the susceptibility to pneumoconiosis. METHODS: Eighteen published case-control studies about TNF-α gene 308, 238 locus polymorphism and pneumoconiosis susceptibility were searched out from sino-foreign databases from January 1994 to December 2010. Meta-analysis was applied on the published research to calculate the pooled OR value (95%CI) and stratified analyze the types and species of pneumoconiosis. RESULTS: Eleven of the published research articles were selected into the analysis, including 10 research focusing on TNF-α gene 308 locus, with 1408 cases and 1639 controls in total. The meta-analysis showed that comparing with Gln/Gln carriers, Arg/Arg, Arg/Gln, Gln/Arg + Arg/Arg carriers were 1.89-fold (95%CI: 1.10 - 3.24), 1.53-fold (95%CI: 1.25 - 1.87), and 1.56-fold (95%CI: 1.28 - 1.90) more susceptible to pneumoconiosis, respectively. The stratified analysis showed that among coal workers, the TNF-α gene 308 locus Arg/Arg, Arg/Gln, Gln/Arg + Arg/Arg carriers were separately 2.29-fold (95%CI: 1.22 - 4.29), 1.56-fold (95%CI: 1.20 - 2.03), 1.64-fold (95%CI: 1.28 - 2.11) more susceptible to pneumoconiosis than Gln/Gln carriers; and among Asian people, the TNF-α gene 308 locus Gln/Arg, Gln/Arg + Arg/Arg carriers were separately 1.58-fold (95%CI: 1.28 - 1.95) and 1.57-fold (95%CI: 1.28 - 1.94) more susceptible to pneumoconiosis than the Gln/Gln carriers. Four case-control research focus on the study of TNF-α gene 238 locus, including 391 cases and 391 controls in total. The analysis showed that comparing with the non-carriers, TNF-α gene 238 locus Arg/Arg, Arg/Gln, Gln/Arg + Arg/Arg carriers were 6.03-fold (95%CI: 1.35 - 26.97), 1.87-fold (95%CI: 1.07 - 3.30) and 2.36-fold (95%CI: 1.37 - 4.07) more susceptible to pneumoconiosis. CONCLUSION: TNF-α gene 308, 238 locus Arg/Arg, Gln/Arg, Gln/Arg + Arg/Arg carriers are more susceptible to pneumoconiosis.

[Diagnostics of atmospheric-pressure radio-frequency capacitively coupled Ar/N2 plasma by optical emission spectroscopy].

An atmospheric-pressure Ar/N2 glow discharge was achieved in a mode stricken between bare metal plate electrodes by radio-frequency power supply at 13.56 MHz. The rotational temperature was determined by using the Boltzmann plot of the OH (A 2sigma+ --> X 2pi) radical and the variation tendency of the gas temperature versus the input power was obtained. Furthermore, the measurement of the sequences of vibrational bands of N2 second positive system (C 3 pi(u) --> B 3 pi(g)) is made and the vibrational temperature was determined correspondingly. The experiment results showed that the emission peaks of N2 (C3 pi(u)) reached the maximum at the nitrogen flow rate of 80 mL x min(-1) with increasing addition of nitrogen, the gas temperature increased from 342 to 523 K when the input power increased from 30 to 210 W, and the vibrational temperature changed slightly when the gas flow rate of nitrogen increased from 30 to 140 mL x min(-1).

Hypoxia inducible factor-1α-dependent epithelial to mesenchymal transition under hypoxic conditions in prostate cancer cells.

Prostate cancer is the most commonly diagnosed cancer in men and the second leading cause of cancer death. Hypoxia is an environmental stimulus that plays an important role in the development and cancer progression especially for solid tumors. The key regulator under hypoxic conditions is stabilized hypoxia-inducible factor (HIF)-1α. In the present study, immune-fluorescent staining, siRNAs, qRT-PC, immunoblotting, cell migration and invasion assays were carried out to test typical epithelial to mesenchymal transition under hypoxia and the key regulators of this process in PC3, a human prostate cancer cell line. Our data demonstrated that hypoxia induces diverse molecular, phenotypic and functional changes in prostate cancer cells that are consistent with EMT. We also showed that a cell signal factor such as HIF-1α, which might be stabilized under hypoxic environment, is involved in EMT and cancer cell invasive potency. The induced hypoxia could be blocked by HIF-1α gene silencing and reoxygenation of EMT in prostate cancer cells, hypoxia partially reversed accompanied by a process of mesenchymal-epithelial reverting transition (MErT). EMT might be induced by activation of HIF-1α-dependent cell signaling in hypoxic prostate cancer cells.

[Long term observation in effects of potassium and calcium supplementation on arterial blood pressure and sodium metabolism in adolescents with higher blood pressure].

OBJECTIVE: To investigate the effects of potassium and calcium supplementation in table salt on reduction of arterial blood pressure and sodium metabolism in adolescents with higher blood pressure. METHODS: A single blind placebo-controlled trial was carried out for two years in 220 adolescents with higher blood pressure, aged 18 - 22 years, who were randomly divided into supplementary group (n = 110) and control group (n = 110). Each of the subjects in the supplementary group and their family members was given 10 mmol of potassium and 10 mmol of calcium mixed in their table salt daily for 24 months. RESULTS: Night urinary sodium and potassium excretion increased (urinary Na(+), P < 0.05; urinary K(+), P < 0.01) and blood pressure lowered by 5.3 mm Hg/1.8 mm Hg in average from the baseline in the supplementary group two years after potassium and calcium supplementation, as compared with that in the control group increased by (1.3/1.7) mm Hg. CONCLUSIONS: Adequate supplement of potassium and calcium in daily table salt intake was an effective way to prevent form hypertension and could promote their urinary sodium excretion and reduction of arterial blood pressure in adolescents with higher blood pressure.