[Characteristics, Sources, and SOAP of VOCs During Winter in Jiyuan].

Haze pollution events often occur in the heavy industry city of Jiyuan. Volatile organic compounds (VOCs) are the precursors of secondary organic aerosol (SOA), which accounts for 15%-20% of particulate matter (PM2.5). Here, PM2.5, O3, VOCs, and trace gases were monitored by using online instruments in Jiyuan from December 1st to 31st. The characteristics, sources, and secondary organic aerosol potential (SOAP) of VOCs were analyzed. The mean concentrations of TVOC were (54.3±27.5)×10-9. Alkanes, halocarbons, and alkynes were the predominant VOCs. The positive matrix factorization model was used to identify and apportion VOCs sources. Eight major sources of VOCs were identified, which included liquefied petroleum gas (LPG) or natural gas (NG), the polyvinyl chloride (PVC) industry, vehicular exhaust, the coking industry, solvent usage, industry, technological process, and fuel evaporation. The SOAP of aromatics was the largest. Among them, BTEXs were the dominant contributors to SOAP.

[Expression levels of Slc7a11 in the skin of Kazakh sheep with different coat colors].

Slc7a11 belongs to solute transporter gene family, encoding cystine/glutamate transporter xCT. It regulates switching between eumelanin and pheomelanin synthesis. In the present study, Real-time PCR was used to detect the mRNA expression levels of Slc7a11 in the skin of Kazakh lambs with different coat colors (black, brown and white), and then the prokaryotic expression plasmid PET-32a-sxCT was constructed to induce the expression of fusion protein. The target pro-tein was purified by Ni-NTA affinity chromatographic separation, and then was used to immunize rabbit in order to produce rabbit anti-sxCT polyclonal antibody. Finally, the expression levels of sxCT were detected in the skin of Kazakh lambs with different hair colors by Western blotting analysis. Results showed that the mRNA expression levels of Slc7a11 differed significantly in the skin of Kazakh lambs with different coat colors, with the highest level in brown coat color, followed by the black, and then the white. The sxCT protein was also detected in the skin of different coat colors by polyclonal antibody, with the highest level in brown coat color, followed by the black, and then the white. It is, therefore, concluded that slc7a11 gene might be associated with the phenotype of coat color in Kazakh sheep.

Differential expressions of MHC-DQB1 mRNA in Chinese merino sheep infected with Echinococosus granuclosus.

The purpose of the present study was to investigate the dynamic changes of MHC-DQB1 mRNA expression in sheep infected with Echinococosus granuclosus. A total of 14 healthy Chinese merino sheep were experimentally infected with E. granuclosus. The blood samples were collected on days 0 (initiation of the infection), 7, 21, 30, and 60 post-infection, respectively. On day 60 post-infection, when the experiment was terminated, all sheep were euthanized to make a diagnosis of cystic echinococcosis (CE) using routine meat inspection and microscopical examination, respectively. The sheep were then divided into two groups according to the diagnostic results: group A (n = 8) consisted of sheep which were diagnosed as CE infection, while group B (n = 6) comprised sheep diagnosed as self-cured or healthy controls. Blood samples obtained during the period of the study were correspondingly divided into groups A and B. The mRNA expression levels of DQB1 revealed significant alterations detected at different stages of E. granuclosus infection in the two groups. Results showed that in group A, DQB1 mRNA expression underwent a progressive increase from day 0 to day 21 post-infection (P = 0.073), and suddenly, suffered from a dramatic drop until day 30 post-infection, and then jumped rapidly and peaked on day 60 post-infection (P = 0.004). Meanwhile, in group B, DQB1 mRNA expression displayed a sharp increase from day 0 to day 7 post-infection (P = 0.000), which thereafter showed a marked decrease until day 30 post-infection, and experienced a plateau from day 30 to day 60 post-infection, remaining at or above that on day 0. It is concluded that DQB1 mRNA expression levels varied in different stages of E. granuclosus infection in sheep. In addition, it appears that the ability to eliminate the parasites possibly depends, at least in part, on the DQB1 expression in the early stage of infection, especially in the first week post-infection.

[The gene ontology and electro localization of ovine skin derived EST-SSR markers].

Abstract: In order to study the potential gene function of ovine EST-SSR markers, nine original EST of Ovine Skin Derived polymorphic EST-SSR loci, which were developed in an early study by our lab, were ontology annotated and Electro localized. The results revealed that the original ESTs of the six loci had high homology with known genes and three of them probably played an important role in wool traits. Compared with its cDNA library, 8 loci were located on chromosomes of cattle. The homology of chromosomes between cattle and sheep was estimated based on the similarity coefficients calculated by positioning markers. Additionally, NJ clustering tree was establishedto serve for electro localization of ovine EST-SSR markers. Finally, 8 EST-SSR markers were successfully positioned on ovine chromosomes. The results from this study not only provide references for further studies on genetic mapping, in silico cloning of key genes for wool traits, but also are helpful to the researchs of chromosome evolution in animal.

Analysis of genetic diversity and phylogenetic relationship of red deer subspecies in XinJiang, China.

Polymorphisms for seven microsatellite loci in three red deer subspecies (9 populations) found in XinJiang were detected by polymerase chain reaction (PCR), 12% nondenaturation polyacrylamide gel electrophoresis and the Sanguinetti silver staining method. Numbers of alleles, average effective numbers of alleles (E) and the average rate of homozygosity, allelic frequencies of seven microsatellite loci, polymorphism information content (PIC), mean heterozygosity (H) and genetic distances among the populations were calculated for each population. Dendrograms were constructed based on genetic distances by the neighbor-joining method (NJ), utilizing molecular evolutionary genetics analysis software PHYLIP (3.6). The phylogenetic tree was constructed based on allelic frequencies using maximum likelihood (ML); the bootstrap value was estimated by bootstrap test in the tree. Lastly, phylogenesis was analyzed. The results showed that four of the seven microsatellite loci were highly polymorphic, but BMS2508 and Celjp0023 showed no polymorphism and BM5004 was a neutral polymorphism. It is our conclusion that the four microsatellite loci are effective DNA markers for the analysis of genetic diversity and phylogenetic relationships among the three red deer subspecies. The mean PIC, H and E-values across the microsatellite loci were 0.5393, 0.5736 and 2.64, which showed that these microsatellite loci are effective DNA markers for the genetic analysis of red deer. C.e. songaricus populations from Regiment 104, 151 and Hami are clustered together. C.e. yarkandensis populations from Regiment 35, Xaya and Alaer are clustered together. These two clusters also cluster together. Lastly, C.e. sibiricus populations from Burqin, Regiment 188 and the first two clusters were clustered together. The phylogenetic relationship among different red deer populations is consistent with the known origin, history of breeding and geographic distributions of populations.

Antibody and cytokine responses to hydatid in experimentally infected Kazakh sheep with hydatidosis resistance haplotype.

Different MHC haplotype of Kazakh sheep has different resistance and susceptibility of hydatidosis. Notably, the MvaIbc-SacIIab-Hin1Iab haplotype of MHC-DRB1 exon two was associated with resistance hydatidosis. In order to analyze the antibody and cytokine responses to hydatidosis in Kazakh sheep with hydatidosis resistance haplotype, eight Kazakh sheep with the haplotype of MvaIbc-SacIIab-Hin1Iab were chosen as the test group, and other eight, which were not associated with hydatidosis resistance or susceptibility, were taken as control. After experimentally infected with hydatid orally, the blood was collected on 0, 7, 14, 30, 45, 60, 75, 90, 105, and 120 days. Serum and mRNA level of the cytokines IL-2, IFN-γ, TNF-α, IL-4, and IL-10 were evaluated by ELISA and fluorescence quantitative real-time polymerase chain reaction, respectively. The total white blood cells and leukomonocytes were determined by automation cytoanalyze. The level of IgE, IgG, and IgM were evaluated by ELISA. The results showed that the total white blood cells and leukomonocytes in test group were significantly higher than in control on 7, 45, 90, and 105 days post-infection (p.i.). The serum level of IL-2 in test group was significantly higher than in control on 45 days p.i., while the difference of IL-2 mRNA expression between test and control group was not significant. The serum level of TNF-α in test group was significantly higher than in control at 90 and 105 days p.i., and the TNF-α mRNA in test group was also significantly higher than in control on 90 days p.i. The level of IgE, IgG, and IgM in test group was higher than in control, but none was significant. The results suggested that the test group, which was predominant of Th1, could induce the protective immunity, while the control, which was predominant of Th2, could induce the susceptibility to infection of hydatidosis.

Identification of SNPs within the sheep PROP1 gene and their effects on wool traits.

Regarding mutations of PROP1 (Prophet of POU1F1) gene significantly associating with combined pituitary hormone deficiency (CPHD) in human patients and animals, PROP1 gene is a novel important candidate gene for detecting genetic variation and growth, reproduction, metabolism traits selection and breeding. The aim of this study was to detect PROP1 gene mutation of the exon 1-3 and its association with wool traits in 345 Chinese Merino sheep. In this study, on the basis of PCR-SSCP and DNA sequencing methods, ten novel SNPs within the sheep PROP1 gene, namely, AY533708: g.45A>G resulting in Glu15Glu, g.1198A>G, g.1341G>C resulting in Arg63Ser, g.1389G>A resulting in Ala79Ala, g.1402C>T resulting in Leu84Leu, g.1424A>G resulting in Asn91Ser, g.1522C>T, g.1556A>T, g.1574T>C, g.2430C>G were reported. In addition, association analysis showed that three genotypes of P4 fragment were significantly associated with fiber diameter in the analyzed population (P=0.044). These results strongly suggested that polymorphisms of the PROP1 gene could be a useful molecular marker for sheep breeding and genetics through marker-assisted selection (MAS).

[Expression of ER, bcl-2, and p53 mRNA in early hybrid embryos of chicken-quail].

Using artificial insemination, 100 female quails were crossed with 10 male chickens. The eggs were collected and hatched in the same incubator. The sex of live hybrid embryos from 66 to 120 hatch hours was determined using multiply PCR of Wpkci. Total 300 male and female embryos at various hatch times were sampled and the relative mRNA abundance of ER, bcl-2, and p53 in the embryos was detected by RT-PCR using beta-actin as the internal standard. The effects of ER, bcl-2, and p53 on the early embryonic development for hybrids between chicken and quail were analyzed. The results showed ER mRNA expression of female hybrids were higher than male hybrids from 66 to 84 hatch hours with a highly significant difference (P<0.01), which indicated that the sex differentiation of hybrids was perhaps happened between 66 to 84 h of embryo stage. The obvious sequential expression of bcl-2 and p53 in the embryonic development indicated that the bcl-2 and p53 genes had an important effect on the development of the hybrid embryos.

[The developmental patterns of GH-R, IGF-1 and IGF-IR gene expression in sheep skin].

Semi-quantitative RT-PCR was applied to investigate the developmental patterns of GH-R, IGF-1 and IGF-IR mRNA expression in skin of two sheep breeds. One breed was the first filial generation (F1) of Romilly Hillys x Merino of China (Xinjiang Agricultural Reclamation line) wool sheep, and the other was Kazak hair sheep. 18S rRNA was used as the internal standard. Sheep were weighed and wool and skin samples were collected at different times. Results showed that body weight increased rapidly during 30-135 days but slowed during 135-255 days. Wool growth increased gradually during 30-135 days, degreased till 180 days of age, but rebounded thereafter. Overall, body weight and developmental patterns of wool growth was not significant different between hair and wool sheep. GH-R mRNA expression in the skin of hair sheep increased significantly during 30-90 days, peaked at 90 days of age (P<0.05), then declined signifi cantly (P<0.05). GH-R mRNA expression in the skin of wool sheep increased significantly until 135 days of age (P<0.01) and then decreased significantly (P<0.01). The peak level was higher in the wool sheep than the hair sheep. The expression of cantly IGF-1 mRNA and IGF-IR mRNA in the skin of hair sheep increased during 30-90 days, then declined significantly (P<0.01). The expression of IGF-1 mRNA and IGF-IR mRNA in the skin of wool sheep were high at birth and then reduced gradually. The IGF-1 mRNA expression in the skin of hair sheep reached its peak at 90 days of age, and was significant higher than that of wool sheep. The expression of GH-R, IGF-1 and IGF-IR mRNA in skin of hair sheep was higher than that of wool sheep before 90 days of age, but was lower after that. The results suggest that GH-R, IGF-1 and IGF-IR mRNA expression in the skin of sheep follows specific developmental patterns, and different patterns exist between the two breeds.

[Polymorphism distributions of 9 microsatellite Loci in Chinese merino sheep].

The polymorphism distributions of nine microsatellite loci, BM6506, BM1824, BM6438, ILSTS004 and OarDB6 on Chinese merino sheep chromosome 1 and BM4621, OarHH55, BM143 and OarJMP8 in sheep chromosome 6 were determined by polymerase chain reaction (PCR) and multiplex gel electrophoresis followed by silver staining. Gene frequency (Pi), power of discrimination (DP), heterozygosity (H), polymorphism information content (PIC) and probability of paternity exclusion (PE) were calculated. All loci obeyed Hardy-Weinberg equilibrium. BM4621 displayed the highest DP, H, PIC and PE values among the nine microsatellite loci. Cumulative DP of the nine microsatellite loci is 0.99999 and cumulative PE is 0.99915. These results showed that the nine microsatellite loci could be used in linkage analysis, individual identification and paternity test in Chinese merino sheep.