[Chemical Characteristics of Shallow Groundwater in the Yellow River Diversion Area of Henan Province and Identification of Main Control Pollution Sources].

The northern plain of Henan in the lower reaches of the Yellow River is an area where the Yellow River is frequently diverted. The shallow groundwater quality in this area is poor， and many types of components have been found to be exceeding the limit value； however， the contribution of various environmental factors to water quality needs to be further quantified. In order to clarify the genesis of water quality of shallow groundwater in the study area， 330 groups of shallow groundwater samples were collected via a regional water quality survey. The evolution of shallow groundwater quality in the Yellow River diversion area of northern Henan was revealed using the principal component-absolute principal component score-multiple linear regression （PCA-APCS-MLR） model. The results showed that the components with a shallow groundwater excess rate greater than 10% in descending order were manganese， iron， total hardness， total dissolved solids， sodium， fluoride， arsenic， chloride ions， sulfate， and ammonium. In particular， the excess rate of manganese reached 76%. The four factors of dissolution enrichment， native origin of soil， redox conditions， and agricultural activities were identified as the main reasons for poor groundwater quality， which accounted for 71.24% of the cumulative interpretation rate of variance. In addition， the recharge from the surface water also influenced the groundwater quality. The effects of dissolution between the water and aquifer matrix and redox condition in the aquifer of the Yellow River dried-riverway like Xinxiang were significantly enhanced， resulting in the increasing concentration of iron， arsenic， total hardness， TDS， and other components in groundwater. Fluoride enrichment was caused by dissolution enrichment， the origin of the soil， and lateral replenishment of the Yellow River. Groundwater with high manganese concentration was widely affected by the soil matrix. Nitrate pollution of the groundwater was caused by the extensive use of chemical fertilizers in agricultural activities in individual areas.

miR-181a Participates in Contextual Fear Memory Formation Via Activating mTOR Signaling Pathway.

Long-term memory formation has been proven to require gene expression and new protein synthesis. MicroRNAs (miRNAs), as an endogenous small non-coding RNAs, inhibit the expression of their mRNA targets, through which involve in new memory formation. In this study, elevated miR-181a levels were found to be responsible for hippocampal contextual fear memory consolidation. Using a luciferase reporter assay, we indicated that miR-181a targets 2 upstream molecules of mTOR pathway, namely, PRKAA1 and REDD1. Upregulated miR-181a can downregulate the PRKAA1 and REDD1 protein levels and promote mTOR activity to facilitate hippocampal fear memory consolidation. These results indicate that miR-181a is involved in hippocampal contextual fear memory by activating the mTOR signaling pathway. This work provides a novel evidence for the role of miRNAs in memory formation and demonstrates the implication of mTOR signaling pathway in miRNA processing in the adult brain.

[Preparation and identification of polyclonal antibody against small peptides of beta-tubulin of Toxoplasma gondii].

The amino acid sequences of beta-tubulin from Toxoplasma gondii stains (GT1 and ME49) and human were aligned by ClustalW2 software. Based on the alignment result, the C-terminal peptides of beta-tubulin of T. gondii were artificially synthesized. Rabbits were immunized with 0.5 mg synthesized peptides for five times at 2-week intervals. Serum samples were collected at the second week after the final immunization, and were analyzed for specific antibodies by ELISA. Finally, the specific-beta-tubulin polyclonal antibody was evaluated by Western blotting with the total protein of RH strain, ME49 strain, and PRU strain of T. gondii, respectively. The results showed that beta-tubulin of T. gondii stains (GT1 and ME49) shared 100% amino-acid sequence identity, and there was 98% amino acid homology between T. gondii and human. The main variable region was the C-terminus. After the fifth immunization, the titers of polyclonal antibody reached 1 : 52,800. Western blotting result indicated that the specific-beta-tubulin polyclonal antibody reacted with beta-tubulin in all the three strains (RH, ME49, and PRU), respectively.

[Preparation and application of the polyclonal antibody of Toxoplasma gondii autophagy protein 8 (TgAtg8)].

OBJECTIVE: To prepare and evaluate specific-TgAtg8 polyclonal antibody. METHODS: The known Saccharomyces cerevisiae Atg protein sequences were used to identify Toxoplasma gondii homologous protein through bioinformatics analysis. TgAtg8 cDNA was amplified and cloned into prokaryotic expression vector pGEX-6p-1. The constructed pGEX-6p-1-TgAtg8 was transformed into E. coli BL21 cells and induced with IPTG for expression. The expression product was analyzed through SDS-PAGE and Western blotting. The recombinant TgAtg8 protein with an N-terminal glutathione-S transferase tag was used to immunize rabbits and raise specific polyclonal antibody against TgAtg8. Subsequently, the antibody was applied for Western blotting and IFA assay. RESULTS: Recombinant expression plasmid of pGEX-6p-1-TgAtg8 was confirmed correct by restriction enzyme digestion and sequencing. SDS-PAGE and Western blotting analysis showed that the recombinant TgAtg8 protein with the predicted molecular weight (M(r)40000) was expressed highly in E. coli BL21. After immunization, the specific antibodies against TgAtg8 protein were produced. The anti-TgAtg8 polyclonal antibody reacted specifically with TgAtg8 fusion protein or endogenous TgAtg8. Importantly, IFA assay determined that the TgAtg8 signal was generally distributed throughout the cytoplasm of the tachyzoites. However, the green fluorescence signal gathered into one or more green spots after induction of autophagy. CONCLUSION: The specific polyclonal antibody against TgAtg8 could be used to observe the dynamics of autophagosome formation in T. gondii, which is useful tool to investigate the autophagic machinery in this parasite.

[Cloning and expression of Toxoplasma gondii autophagy-related protein 3 gene and preparation of its polyclonal antibody].

OBJECTIVE: To clone and express autophagy-related protein 3 (TgAtg3) gene of Toxoplasma gondii, and obtain the specific polyclonal antibody against TgAtg3. METHODS: TgAtg3 cDNA was inserted into prokaryotic expression vector pET28a. After identification, the constructed plasmid pET28a-TgAtg3 was transformed into E. coli Rosetta cells, and induced by special induction medium for expression of the protein. The recombinant protein was purified via Ni-NTA affinity chromatography. Western blotting assay was performed with anti-His tag mouse monoclonal antibody as the primary antibody. Rabbits were immunized with 125 µg purified TgAtg recombinant protein. Each rabbit received 4 immunizations at 2-week intervals with the same dose of antigen. The specific anti-TgAtg3 polyclonal antibody was obtained, and analyzed by Western blotting and indirect immunofluorescence assay (IFA). RESULTS: pET28a-TgAtg3 plasmid was identified by restriction enzyme digestion, PCR amplification and sequencing. SDS-PAGE and Western blotting analysis showed that the recombinant TgAtg3 protein (about Mr 44,000) was expressed in E. coli Rosetta cells. TgAtg3 protein from tachyzoite lysates was recognized by the specific anti-TgAtg3 polyclonal antibody. IFA assay determined that the specific polyclonal antibody bound to TgAtg3 protein from the cytoplasm of tachyzoites. CONCLUSION: The obtained soluble polyclonal antibody against TgAtg3 can specifically react to the endogenous TgAtg3 protein.

8-{[3-(3-Meth-oxy-phen-yl)-1,2,4-oxa-diazol-5-yl]meth-oxy}quinoline monohydrate.

In the title hydrate, C19H15N3O3·H2O, the three aromatic groups in the quinoline derivative are close to coplanar: the central oxa-diazole fragment makes dihedral angles of 15.7 (2)° with the benzene ring and 5.30 (14)° with the quinoline ring system. In the crystal, the organic mol-ecules are connected with water mol-ecules by pairs of O-H⋯N hydrogen bonds involving the quinoline and oxa-diazole N atoms. The mol-ecules form stacks along the a axis, neighboring mol-ecules within each stack being related by inversion and the shortest distance between the centroids of the oxa-diazole and pyridine rings being 3.500 (2) Å. Mol-ecules from neighboring stacks are linked by weak C-H⋯O hydrogen bonds, forming a three-dimensional structure.

2-tert-Butyl-1-(4-nitro-amino-1,2,5-oxadiazol-3-yl)diazene 1-oxide.

In the title compound, C(6)H(10)N(6)O(4), the nitro-amine -NHNO(2) substituent and the C-N=N(→ O) unit of the other substituent of the oxadiazole ring are nearly coplanar with the five-membered ring [dihedral angles = 5.7 (1) and 3.0 (1)°]. The amino group of the -NHNO(2) substituent is a hydrogen-bond donor to the two-coordinate N atom of the C-N=N(→ O) unit.

[Molecular mapping of the gene(s) controlling petal-loss trait in Brassica napus].

An F2 population derived from a cross between apetalous line' APT02 'and normal petalled cultivar 'ZS NO.4' was used for molecular marker searching and chromosomal mapping of the gene(s) controlling petal-loss trait in Brassica napus. Twenty pairs of AFLP primers and 170 pairs of SRAP primers were selected and screened from two parents. In further selection through bulked segregant analysis (BSA) approach, one SRAP marker e8m3_4 (600 bp) and one AFLP marker E3247_15 (150 bp) were obtained and found to be linked to the gene(s) controlling petal-loss trait, with the genetic distance of 5 cM and 13.5 cM. A linkage map in Brassica napus was constructed. It consisted of 213 AFLP56 SSR loci and a morphology marker throughout 17 main linkage groups, two triplet and four linkage pairs. Total length of the map covered 2,487.1 cM, and average interval between markers was 10.09 cM. By genetic mapping, the gene(s) controlling petal-loss trait (WHB)was mapped in LG4.