Nonpoint Source Pollution (NPSP) Induces Structural and Functional Variation in the Fungal Community of Sediments in the Jialing River, China.

Nonpoint source pollution (NPSP) from human production and life activities causes severe destruction in river basin environments. In this study, three types of sediment samples (A, NPSP tributary samples; B, non-NPSP mainstream samples; C, NPSP mainstream samples) were collected at the estuary of the NPSP tributaries of the Jialing River. High-throughput sequencing of the fungal-specific internal transcribed spacer (ITS) gene region was used to identify fungal taxa. The impact of NPSP on the aquatic environment of the Jialing River was revealed by analysing the community structure, community diversity, and functions of sediment fungi. The results showed that the dominant phylum of sediment fungi was Rozellomycota, followed by Ascomycota and Basidiomycota (relative abundance > 5%). NPSP caused a significant increase in the relative abundances of Exosporium, Phialosimplex, Candida, Inocybe, Tausonia, and Slooffia, and caused a significant decrease in the relative abundances of Cercospora, Cladosporium, Dokmaia, Setophaeosphaeria, Paraphoma, Neosetophoma, Periconia, Plectosphaerella, Claviceps, Botrytis, and Papiliotrema. These fungal communities therefore have a certain indicator role. In addition, NPSP caused significant changes in the physicochemical properties of Jialing River sediments, such as pH and available nitrogen (AN), which significantly increased the species richness of fungi and caused significant changes in the fungal community ß-diversity (P < 0.05). pH, total phosphorus (TP), and AN were the main environmental factors affecting fungal communities in sediments of Jialing River. The functions of sediment fungi mainly involved three types of nutrient metabolism (symbiotrophic, pathotrophic, and saprotrophic) and 75 metabolic circulation pathways. NPSP significantly improved the pentose phosphate pathway, pentose phosphate pathway, and fatty acid beta-oxidation V metabolic circulation pathway functions (P < 0.05) and inhibited the chitin degradation to ethanol, super pathway of heme biosynthesis from glycine, and adenine and adenosine salvage III metabolic circulation pathway functions (P < 0.05). Hence, NPSP causes changes in the community structure and functions of sediment fungi in Jialing River and has adversely affected for the stability of the Jialing River Basin ecosystem.

[In vitro killing of adult Trichinella spiralis by exogenous nitric oxide].

OBJECTIVE: To investigate the lethal effect of exogenous nitric oxide on adult worms of Trichinella spiralis in vitro. METHODS: Adult worms of T. spiralis isolated from the small intestine of Trichinella-infected BALB/c mice were cultured in RPMI 1640 medium with sodium nitroprusside (SNP) in different final concentration of 0, 0.02, 0.05, 0.10, 0.20, 0.50, and 1.00 mmol/L, 1.00 mmol/L SNP+0.15 mmol/L hemoglobin (Hb), 1.00 mmol/L SNP+0.15 mmol/L FeSO4, 1.00 mmol/L SNP+1.00 mmol/L L-cysteine (L-cyst), 1.00 mmol/L SNP+0.15 mmol/L FeSO4 +1.00 mmol/L L-cyst, respectively, and incubated at 37 degrees C in a humidified 5% CO2 atmosphere. On the 4th day after incubation, the adult worms were stained with safranin, and observed under light microscope. The worm mortality in the groups was analyzed. RESULTS: Under concentration of 0.02 and 0.05 mmol/L, SNP was not cytotoxic to adult T. spiralis with an inhibition of (1.4 +/- 1.2)% and (3.2 +/- 1.0)%, respectively. The worm mortality in the groups of SNP 0.10, 0.20, 0.50, and 1.00 mmol/L was (9.9 +/- 1.8)%, (37.7 +/- 2.5)%, (50.1 +/- 3.5)%, and (80.8 +/- 1.1)%, respectively, significantly higher than that of negative control group [(1.9 +/- 0.2)%, P < 0.05]. There was a positive linear correlation between the worm mortality and SNP concentration in the range of 0.02-1.00 mmol/L. Combination of hemoglobin, L-cyst, FeSO4 and FeSO4+L-cyst with 1.00 mmol/L SNP led to a decrease of the mortality from (80.8 +/- 1.1)% to (56.5 +/- 3.7)%, (69.8 +/- 2.3)%, (74.8 +/- 2.4)%, (72.7 +/- 5.6)%, respectively. CONCLUSION: Exogenous nitric oxide released from SNP can kill adult worms of Trichinella spiralis. However, hemoglobin and L-cysteine+FeSO4 can reverse its lethal effect on the parasites.

[Effect of Different Land Use Types on the Diversity of Soil Bacterial Community in the Coastal Zone of Jialing River].

The purpose of this study was to clarify the differences in soil bacterial community diversity in different land uses in the coastal zone of the Jialing River and to provide theoretical reference for reasonable land development and resource protection of the Jialing River basin. Based on second generation high-throughput sequencing technology, the diversity, structural composition, functional types, and influencing factors of soil bacterial communities in different land use types were analyzed. The results showed that:① there were some differences in soil physical and chemical properties among different land uses. The nutritional indexes of agricultural land and soil wetland were significantly higher than those of natural forest land and sandy soil wetland (P<0.05). ②The Ace, Shannon, and Chao indexes of sandy soil wetland and soil wetland were significantly higher than those of natural forest land and agricultural land (P<0.05). The Simpson index of soil bacteria in different land use types was significantly different (P<0.05). ③ The dominant phyla of soil bacteria in the coastal zone of the Jialing River were Proteobacteria, Actinobacteriata, Acidobacteriata, and Chloroflexi, and the relative abundance accounted for more than 65%. The community structure of sandy soil wetland was similar to that of soil wetland and was significantly different from that of other land use types (P<0.05). ④ Primary functional metabolism was the core function of the bacterial community, accounting for more than 75%. Agricultural activities had the greatest impact on the community function, which significantly improved the metabolism of amino acids and carbohydrates, enhanced the utilization of nitrogen and phosphorus, reduced the metabolism of energy and coenzyme vitamins, and weakened the resistance of bacterial community to environmental impact (P<0.05). ⑤ Total phosphorus (TP) was the key influencing factor of the soil bacterial community. In conclusion, the construction of the wetland habitat in the Jialing River coastal zone is most conducive to the development of underground ecological diversity and the stability of the soil ecosystem.

Over-expression of vitronectin correlates with impaired survival in gastric cancers.

ABSTRACT: Over-expression of vitronectin (VN) is associated with tumorigenesis. The present study aimed to evaluate the prognostic value of VN expression in gastric cancer.The least absolute shrinkage and selection operator analysis was performed to screen the hub gene from The Cancer Genome Atlas gastric cancer patients with complete follow-up data, and 347 patients were finally included. Moreover, 102 patients were enrolled from the Affiliated Fuzhou First Hospital of Fujian Medical University. VN expression in paired gastric cancer and adjacent gastric normal tissues was detected using immunohistochemistry, and the clinicopathological significance of VN expression was evaluated. The prognostic significance of VN expression in gastric cancer patients was evaluated using by Kaplan-Meier method and Cox regression analysis and confirmed using Oncomine.VN was the prognosis relative gene which screened by The Cancer Genome Atlas dataset. Moreover, we identified the VN expression in an external dataset by immunohistochemistry. The result demonstrated that VN expression was remarkedly elevated in gastric cancer tissues (P < .001). High VN expression correlated with higher pathological Tumor-Node-Metastasis stage, and poorer survival outcomes. Cox regression analysis showed that VN expression was independently predictive of overall survival (OS) and disease-free survival (P = .004, P < .001, respectively). A prognostic risk score for OS was built based on VN expression. A meta-analysis from Oncomine datasets revealed that significantly lower VN mRNA levels in gastric cancer correlated with poorer OS.VN expression could be a prognostic marker of gastric cancer.

[Deltamethrin resistance, metabolic detoxification enzyme and kdr mutation in Anopheles sinensis in region along Huaihe River in Anhui Province].

OBJECTIVE: To understand the deltamethrin resistance level, GTS and P450 metabolic detoxification enzyme activities, and mutations in the para-type sodium channel gene that confers knockdown resistance (kdr) in Anopheles sinensis mosquitoes from malaria endemic areas along the Huaihe River in Anhui Province, China. METHODS: An. sinensis adult mosquitoes were collected in Lilou, Mohekou and Tuohu townships of Bengbu City, Anhui Province from August to September, 2011. The insecticide resistance bioassays were performed on adult mosquitoes by using the standard WHO susceptibility test with diagnostic concentrations of deltamethrin 0.05%, and the mosquito resistance status was classified based on the WHO resistance classification criteria. The metabolic detoxification enzyme activities were measured in randomly selected mosquitoes, and the IIS6 region of the para-type sodium channel gene was amplified by PCR and sequenced to detect mutations at the codon 1014. RESULTS: The knockdown rates within 60 min exposure to deltamethrin test paper were 4.1%, 7.0% and 8.2%, and the mortality rates were 8.2%, 12.0% and 12.8% for mosquitoes collected from Lilou, Mohekou and Tuohu townships, respectively. These three populations were classified as highly resistant populations based on the WHO resistance classification criteria. The GST and P450 enzyme activities of the three populations were significantly higher than those of the susceptible laboratory population (P < 0.001). L1014C and L1014F mutations were detected, and the wild type homozygote kdr genotype was not found. These three populations exhibited a small but insignificant difference in kdr allele frequencies. No mutation was found in the laboratory susceptible mosquitos. CONCLUSION: The An. sinensis mosquito populations from the regions along the Huaihe River in Anhui Province are strongly resistant to pyrethroid insecticides, and exhibit significantly higher metabolic detoxification enzyme activities than the laboratory susceptible population. The high frequency of kdr mutation is identified in the mosquito populations from the region along the Huaihe River in Anhui Province, China.

[Construction, identification and immunoreactivity of Mycobacterium tuberculosis DNA vaccine pVAX1/ESAT-6 plasmid].

OBJECTIVE: To construct a Mycobacterium tuberculosis DNA vaccine pVAX1/ESAT-6 plasmid and investigate its immunoreactivity. METHODS: The ESAT-6 gene fragment amplified from Mycobacterium tuberculosis genome was inserted into pVAX1 vector to construct the recombinant plasmid pVAX1/ESAT-6, which was identified by restriction enzyme digestion and sequencing. The recombinant plasmid was transformed into Hela cells using Sofast® Transfection reagent, and the cellular expressions of ESAT-6 mRNA and protein were analyzed by RT-PCR and immunofluorescence assay, respectively. The recombinant plasmid pVAX1/ESAT-6 was also transfected into mouse by electronic pulse method, and the mouse serum IFN-γ level and anti-ESAT-6 IgG antibody level were detected by ELISA, mouse lymphocyte proliferation assessed with flow cytometry, and IFN-γ-secreting lymphocytes counted using ELISPOT. RESULTS: Double restriction-enzyme digestion and sequencing showed that the inserted fragment in the recombinant plasmid pVAX1/ESAT-6 was identical to ESAT-6 gene with an inframe insertion. RT-PCR yielded the target band as expected on agarose gel, and immunofluorescence assay of the transfected cells showed specific green fluorescence signals. The mice transfected with the recombinant plasmid showed significantly elevated serum level of anti-ESAT-6 IgG antibody and increased serum IFN-γ level, spleen cell proliferation, and number of IFN-γ-secreting lymphocytes. CONCLUSION: The Mycobacterium tuberculosis DNA vaccine pVAX1/ESAT-6 plasmid we constructed can induce high levels of cellular and humoral immunoreactivity in mice.