The Competitive Endogenous RNA (ceRNA) Regulation in Porcine Alveolar Macrophages (3D4/21) Infected by Swine Influenza Virus (H1N1 and H3N2).

H1N1 and H3N2 are the two most common subtypes of swine influenza virus (SIV). They not only endanger the pig industry, but are also a huge risk of zoonotic diseases. However, the molecular mechanism and regulatory network of pigs (hosts) against influenza virus infection are still unclear. In this study, porcine alveolar macrophage cell (3D4/21) models infected by swine influenza virus (H1N1 and H3N2) were constructed. The expression profiles of miRNAs, mRNAs, lncRNAs and circRNAs after H1N1 and H3N2 infected 3D4/21 cells were revealed in this study. Then, two ceRNAs (TCONS_00166432-miR10391-MAN2A1 and novel_circ_0004733-miR10391-MAN2A1) that regulated H1N1 and H3N2 infection in 3D4/21 cells were verified by the methods of bioinformatics analysis, gene overexpression, gene interference, real-time quantitative PCR (qPCR), dual luciferase activity assay and RNA immunoprecipitation (RIP). In addition, the important candidate molecules (miR-10391, TCONS_00166432, and novel_circ_0004733) were identified by qPCR and enzyme linked immunosorbent assay (ELISA). Finally, the regulatory effect and possible molecular mechanism of the target gene MAN2A1 were identified by the methods of gene interference, qPCR, Western blot and ELISA. The results of this study suggested that TCONS_00166432 and novel_circ_0004733 could competitively bind miR-10391 to target the MAN2A1 gene to regulate swine influenza virus infecting 3D4/21 cells. This study reported for the first time the ceRNA networks involved in the regulation of the swine influenza virus infecting 3D4/21 cells, which provided a new insight into the molecular mechanism of 3D4/21 cells against swine influenza virus infection.

Removal of Cr (VI) by Biochar Derived from Six Kinds of Garden Wastes: Isotherms and Kinetics.

Garden waste is one of the main components of urban solid waste which affects the urban environment. In this study, garden waste of Morus alba L. (SS), Ulmus pumila L. (BY), Salix matsudana Koidz (LS), Populus tomentosa (YS), Sophora japonica Linn (GH) and Platycladus orientalis (L.) Franco (CB) was pyrolyzed at 300 °C, 500 °C, 700 °C to obtain different types of biochar, coded as SSB300, SSB500, SSB700, BYB300, etc., which were tested for their Cr (VI) adsorption capacity. The results demonstrated that the removal efficiency of Cr by biochar pyrolyzed from multiple raw materials at different temperatures was variable, and the pH had a great influence on the adsorption capacity and removal efficiency. GHB700 had the best removal efficiency (89.44%) at a pH of 2 of the solution containing Cr (VI). The pseudo second-order kinetics model showed that Cr (VI) adsorption by biochar was chemisorption. The Langmuir model showed that the adsorption capacity of SSB300 was the largest (51.39 mg·g-1), BYB500 was 40.91 mg·g-1, GHB700, CBB700, LSB700, YSB700 were 36.85 mg·g-1, 36.54 mg·g-1, 34.53 mg·g-1 and 32.66 mg·g-1, respectively. This research, for the first time, used a variety of garden wastes to prepare biochar, and explored the corresponding raw material and pyrolysis temperature for the treatment of Cr (VI). It is hoped to provide a theoretical basis for the research and utilization of garden wastes and the production and application of biochar.

Does tourism industry agglomeration reduce carbon emissions?

The global level of climate change agreement and the extensive development of China's industrialization process have caused China to face severe pressures regarding energy conservation and emission reduction. Tourism industry plays an important role in promoting steady economic growth and improving ecological environment in China. The agglomeration economic effect it produces can reduce carbon emissions, but the crowding effect may not be conducive to low carbon development. Therefore, how to reduce carbon emissions while promoting the development of tourism industry has become an urgent problem. This paper measures the level of tourism industry agglomeration and carbon emission in 30 regions of China and uses the method of spatial econometrics to analyze the impact of tourism industry agglomeration on carbon emissions. The empirical results prove that tourism industry agglomeration can reduce the carbon emissions of local and neighboring regions in China. However, this does not mean that the larger the scale of tourism industry agglomeration, the more conducive to the reduction of carbon emissions. For developed tourism groups, the impact of agglomeration effect on carbon emissions of local and adjacent areas shows a U-shaped relationship. When the agglomeration level exceeds 1.963, the expansion of agglomeration scale will increase local carbon emissions. For underdeveloped tourism areas, there is an inverted U-shaped relationship between agglomeration level and carbon emissions both in local and adjacent areas. When the agglomeration level reaches 1.883, the expansion of agglomeration scale will reduce local carbon emissions. The conclusions provide a basis for Chinese government to guide the low-carbon development of the tourism industry from another perspective.

Enhanced Production of Active Ecumicin Component with Higher Antituberculosis Activity by the Rare Actinomycete Nonomuraea sp. MJM5123 Using a Novel Promoter-Engineering Strategy.

Ecumicins are potent antituberculosis natural compounds produced by the rare actinomycete Nonomuraea sp. MJM5123. Here, we report an efficient genetic manipulation platform of this rare actinomycete. CRISPR/Cas9-based genome editing was achieved based on successful sporulation. Two genes in the ecumicin gene cluster were further investigated, ecuN and ecuE, which potentially encode a pretailoring cytochrome P450 hydroxylase and the core peptide synthase, respectively. Deletion of ecuN led to an enhanced ratio of the ecumicin compound EcuH16 relative to that of EcuH14, indicating that EcuN is indeed a P450 hydroxylase, and there is catalyzed hydroxylation at the C-3 position in unit12 phenylalanine to transform EcuH16 to the compound EcuH14. Furthermore, promoter engineering of ecuE by employing the strong promoter kasO*P was performed and optimized. We found that integrating the endogenous ribosome-binding site (RBS) of ecuE together with the RBS from kasO*P led to improved ecumicin production and resulted in a remarkably high EcuH16/EcuH14 ratio. Importantly, production of the more active component EcuH16 was considerably increased in the double RBSs engineered strain EPR1 compared to that in the wild-type strain, reaching 310 mg/L. At the same time, this production level was 2.3 times higher than that of the control strain EPA1 with only one RBS from kasO*P. To the best of our knowledge, this is the first report of genome editing and promoter engineering on the rare actinomycete Nonomuraea.

[Soil physical properties of different hedgerow systems in upper reaches of Yangtze River].

Based on the investigation of present hedgerows in the upper reaches of Yangtze River, this paper analyzed the soil physical properties at different positions of three kinds of hedgerows (arbor, shrub, and grass). Comparing with those between the hedgerows, the soil physical properties within the hedgerows improved significantly. The average values of soil porosity, moisture content, saturated conductivity, water stable aggregates content, anti-erodibility index, anti-scouribility index, and clay content within the arbor, grass, and shrub hedgerows increased by 18.8%, 30.1%, 12.9%, 139.3%, 108.3%, 95.9%, and 25.5%, and the soil bulk density and sand content averagely decreased by 17.3% and 9.6%, respectively. The soil properties within the three hedgerows differed significantly. The soil anti-scouribility index within arbor hedgerow was the highest; the soil porosity, moisture content, saturated conductivity, anti-scouribility index, water-stable aggregates content, and clay content within the shrub hedgerow were higher than those within the tree and grass hedgerows; while the soil bulk density within the shrub hedgerows was lower than that within the tree and grass hedgerows. Because of the differences in the affecting degree of hedgerow on the soil physical properties at different positions of the three hedgerow systems, the related parameters presented definite horizontal variation at steep lands within, before, and behind the hedgerows, and between the hedgerows. The coefficient of variation (CV) of soil moisture content, anti-erodibility index, saturated conductivity, and clay content of arbor hedgerows was bigger than that of shrub and grass hedgerows, while the CV of soil bulk density, porosity, water-stable aggregates content, and anti-scouribility index of shrub hedgerow was bigger than that of arbor and grass hedgerows.

Comparative genomic analysis of growth hormone gene in geese.

To explore the mutation characteristic of growth hormone (GH) gene in geese, all the exons and introns of the gene were amplified by 20 pairs of primers, and then single nucleotide polymorphisms (SNPs) were detected by single strand conformation polymorphism (SSCP) and subsequently confirmed by sequencing. There were six SNPs per 1000 nucleotides in exons compared to two SNPs per 1000 nucleotides in intron regions. The variant in exons contained only one non-synonymous mutation and three synonymous mutations. The results show that its sequence identity with chicken and duck were 77.54% and 92.38%, respectively, which may be concluded that the GH gene was highly conservative in phylogenesis, although there were differences between waterfowls and chicken in their evolution process.

[Macropore characteristics and its relationships with the preferential flow in broadleaved forest soils of Simian Mountains].

Brilliant blue dyeing and water breakthrough curve were applied to study the number and distribution of macropores and their relations to the preferential flow in typical sub-tropic broad-leaved forest soils of Simian Mountains. The radii of the macropores were mainly between 0. 3 and 3.0 mm, with the macroporosities in the range of 6.3% to 10.5%, and the macropores were always distributed in aggregation with increasing soil depth. The number of the macropores in each radius interval of dye-stained areas was tenfold increase than that of blank areas. The number of the macropores with radius larger than 0.3 mm, especially larger than 1.5 mm, was the most important factor affecting the occurrence of preferential flow. Significant correlations were found between the number of macropores and the water steady effluent volume, with the highest correlation coefficients of 0.842 and 0.879 for the radii intervals of 0.7-1.5 mm and 1.5-3.0 mm, respectively. Macro-pore continuity in dye-stained areas was better than that in blank areas, especially in the radius interval of 1.5-3.0 mm, with the biggest difference of 78.31%. In dye-stained areas, the number of macropores decreased gradually with soil depth. The filler-like distribution of macropores formed an effective water pressure gradient, which resulted in the preferential transport of water.

[Cloning and polymorphism analysis of intron 2 of the GH gene in Takifugu].

Size and nucleotide polymorphisms of intron 2 of the growth hormone (GH) gene was studied in 82 individuals from Takifugu obscurus, Takifugu rubripes and Takifugu niphobles with PAGE, SSCP and cloning. Results showed that there were nine length variants (A~I) in these fishes, ranging from 271351 bp, with a frequency of variation of 24.22%. Sequence analysis of the 9 length variants showed the average percentages of the four bases (ATG and C) were 17.15, 20.77, 37.38 and 24.70, respectively, and the difference between GC content (45.47) and AT content (54.53) was not remarkable. Length variation was mainly due to the variable number of microsatellite TCTG repeats (N=20 approximately 40). Further sequence comparison revealed 4 substitution sites: a (C-->A) transversion at nucleotide 103 and 3 transitions 83(C-->T), 101(A-->G) and 296(G-->A), respectively. Phylogenetic analysis of these length variants placed DD and II into one group first, which was subsequently joined by GG and AA, BB and CC, EE and HH were then clustered together, FF was joined last. Thus, intraspecific variations were much smaller than interspecific variations for the intron 2 of the GH gene.

[Analysis of polymorphisms in exon 14 in porcine Mx1 gene and identification of new mutation points].

By the PCR-RFLP method, the polymorphisms of exon 14 of Mx1 gene were detected in 7 native and foreign pig breeds. Taken together with the analysis of restriction enzyme Hin6 I digestion, there were 6 genotypes and 3 alleles. And, all individuals of Duroc exhibited the AA genotype but the Sutai pigs exhibited all three kinds of genotypes. The BB genotype was detected only in Meishan pigs and their derivate Sutai pigs. Among all pig breeds, the allele B only appeared in Chinese indigenous pig breeds and developed pig breed in this study, and the allele A was dominant allele in all pig breeds except for Songliao black pig. The results of Chi Square test showed that there were abundant polymorphisms in all pig breeds. The gene frequency of Meishan pig and Songliao black pig showed greatly significant difference to other pig breeds (P<0.01), Sutai pig showed greatly significant difference (P<0.01) to other pig breeds except for Pietrain; and Huai pig showed greatly significant differences (P<0.01) with Pietrain and other indigenous Chinese pig breeds, but no difference with Duroc and Yorkshire (P>0.05). Three new mutation points in three genotypes were identified by sequencing comparison analysis of PCR products, two of which resulted in Thr-->Ala and Glu-->Arg respectively, the last one was a silence mutation, the Glu-->Arg mutation point and this silence mutation point only appeared in the individuals with the BB genotype.

[Polymorphic analysis of intron 2 and 3 of growth hormone gene in duck].

The single nucleotide polymorphism (SNP) of growth hormone gene was investigated in various breeds of duck, including Beijing ducks, Xihu mallards, Jinding ducks, Shan Partridge ducks, Jingjiang ducks and Shaoxing ducks. The primers for intron 2 and 3 in GH gene were designed based on the database of duck genomic sequence and the SNPs were detected by PCR-SSCP method. Eight SNPs were found among individuals within a breed, which were 2593(C-T), 2770(G-A), 2813(T-A), 2829(C-A), 2894(C-T), 2896(T-C), and 3100(C-G) in intron 2 and 3270(A-G) in intron 3. The analytic results showed that the frequencies of genotypes in different breeds were significantly different. Based on these SNPs, Beijing ducks and Shaoxing ducks represented their own unique conservativeness, indicating that these SNPs may have relationship with some productive traits of duck.

[The study on the technique of sexing bovine pre-implantation embryos with two-temperature gradient multiplex PCR].

For the stable, reliable, fast method of sexing bovine pre-implantation embryos is still play very important role in husbandry production, the amplification experiment under the condition of the two-temperature gradient PCR was done with bovine samples such as genome, cloning embryos, embryos respectively. As a result, the stable, simple, fast method of two-temperature gradient PCR for sexing bovine pre-implantation embryos was obtained, which only took 57 minutes to identify the embryo sex. A total of 30 dairy embryos were sexed with the two-temperature gradient PCR method in the study. 15 identified embryos (11 female, 4 male) were transferred into 15 recipient cows, among them 7 were pregnant after 60 days. In the end, 5 female calves were aborted in late pregnancy, and 1 female and 1 male dairy calf were born. The sexes of aborted and born calves were fully in accordance with the embryo sex predetermination with PCR method.

[Progress in the study on genetic mechanism for high prolificacy in farm animals.].

Reproduction ability of farm animals is a quantitative trait that affects largely the husbandry productions. Progress in the study on genetic mechanism for high prolificacy in farm animals were reviewed at genome level and mRNA expression level in this paper. All these research results indicate that high prolificacy is a trait affected by a lot of genes, that is, the co-expression and interaction of these genes result in the formation of the trait. A lot of progresses have been made at all these two research levels. But differing from the genome level, the research at mRNA expression level only involves the genes expressed at special time in special tissues, and the genes directly contributed to the formation of traits. The genetic mechanism for this trait has been studying at the levels of genome and mRNA expression nowadays. With the development of sciences, new research methods for the trait will be provided, but only the combination of them with all these research levels will be the most effective.