Comparison of Characteristics and Differences in Early Immune Organ Development in Different Strains of Tianfu Broiler

To effectively develop and utilize high-quality Tianfu broilers, this study evaluated the morphological and structural characteristics of the immune organs of such broilers with different strains (HS1 and HS2) at different developmental stages and analyzed the distribution of mast cells by toluidine blue staining. Moreover, the localization and expression of immunoglobulin, complement C3, C4 and CD3 in immune organs were also detected. The results showed that although there was no significant difference in the development of immune organs in the HS1 and HS2, the number of lymphatic follicles and capsule thickness in the spleen and bursa of Fabricius in HS1 were greater than those in HS2. Additionally, the number of mast cells in the spleen of HS1 was greater at Day 1 and Day 21 and was significantly higher than that of HS2 (p<0.05); the number of mast cells in the bursa of Fabricius reached 9.17 on Day 7, which was significantly higher than that of HS2 (p<0.05). Moreover, the serum IgA and IgM levels in HS1 were higher than those in HS2 on Day 14 and 21 (p<0.05). In addition, the complement C3 content in HS1 was significantly or extremely significantly higher than that in HS2 on Days 1, 14 and 21 (p<0.01, p<0.05), respectively, but significantly lower than in HS2 on Day 7 (p<0.05). These results indicated that the disease resistance of the HS1 line was stronger than that of the HS2 line, which lays a foundation for future disease- resistance breeding of Tianfu broilers.(AU)

Genetic Diversity of Mitochondrial DNA of Chinese Black-bone Chicken

The black-bone chicken has special economic value in Chinese poultry breeds, which also are valued for the medicinal properties of their meat in traditional Chinese medicine. In order to protect the genetic resources of native black-bone chicken breeds, we analyzed the genetic diversity and matrilineal components of 64 mtDNA D-loop partial sequences from three native black-bone chicken breeds, together with reported 596 black-bone chicken mtDNA sequences from China, Japan, and Korea. A total of 108 haplotypes were observed from 73 variable sites. These domestic chicken mtDNA sequences could be assigned into seven clades (A-G). The results indicated that 71.97% of the black-bone haplotypes were related to the reference sequence that may originate from Eurasia, while the minor part of mtDNA sequences presumably derive from Southeast Asia, China, and Japan. Three clades were shared by Korean, Japanese, and Chinese black-bone chickens. These results provide basic data useful for making new breeding and conservation strategies for the black-bone chicken in China.(AU)

Genetic Diversity of Mitochondrial DNA of Chinese Black-bone Chicken

The black-bone chicken has special economic value in Chinese poultry breeds, which also are valued for the medicinal properties of their meat in traditional Chinese medicine. In order to protect the genetic resources of native black-bone chicken breeds, we analyzed the genetic diversity and matrilineal components of 64 mtDNA D-loop partial sequences from three native black-bone chicken breeds, together with reported 596 black-bone chicken mtDNA sequences from China, Japan, and Korea. A total of 108 haplotypes were observed from 73 variable sites. These domestic chicken mtDNA sequences could be assigned into seven clades (A-G). The results indicated that 71.97% of the black-bone haplotypes were related to the reference sequence that may originate from Eurasia, while the minor part of mtDNA sequences presumably derive from Southeast Asia, China, and Japan. Three clades were shared by Korean, Japanese, and Chinese black-bone chickens. These results provide basic data useful for making new breeding and conservation strategies for the black-bone chicken in China.

Correction to: Role of intestinal microbiome in American ginseng-mediated colon cancer prevention in high fat diet-fed AOM/DSS mice.

This article was originally published with the wrong title.

Role of intestinal microbiome in American ginseng-mediated colon cancer prevention in high fat diet-fed AOM/DSS mice [corrected].

OBJECTIVE: Chronic intestinal inflammation is a risk factor for colorectal cancer (CRC) initiation and development. Diets that are rich in Western style fats have been shown to promote CRC. This study was conducted to investigate the role of intestinal microbiome in American ginseng-mediated CRC chemoprevention in a mouse model. The population and diversity of enteric microbiome were evaluated after the ginseng treatment. METHODS: Using an azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced gut inflammation and tumorigenesis mouse model, the effects of oral American ginseng on high fat diet-associated enteric pathology were determined. After establishment of a 16S rRNA illumina library from fecal samples, MiSeq sequencing was carried out to reveal the microbial population. The alpha and beta diversities of microbiome were analyzed. RESULTS: American ginseng significantly attenuated AOM/DSS-induced colon inflammation and tumorigenesis by reducing the colitis score and colon tumor multiplicity. The MiSeq results showed that the majority of sequences fell into three phyla: Firmicutes, Bacteroidetes and Verrucomicrobia. Further, two significant abundance shifts at the family level, Bacteroidaceae and Porphyromonadaceae, were identified to support ginseng's anti-colitis and anti-tumor effects. In addition, alpha and beta diversity data demonstrated that ginseng led to a profound recovery from the AOM/DSS-induced dysbiosis in the microbial community. CONCLUSION: Our results suggest that the CRC chemopreventive effects of American ginseng are mediated through enteric microbiome population-shift recovery and dysbiosis restoration. Ginseng's regulation of the microbiome balance contributes to the maintenance of enteric homeostasis.

Genetic Diversity of Mitochondrial DNA of Chinese Black-bone Chicken

ABSTRACT The black-bone chicken has special economic value in Chinese poultry breeds, which also are valued for the medicinal properties of their meat in traditional Chinese medicine. In order to protect the genetic resources of native black-bone chicken breeds, we analyzed the genetic diversity and matrilineal components of 64 mtDNA D-loop partial sequences from three native black-bone chicken breeds, together with reported 596 black-bone chicken mtDNA sequences from China, Japan, and Korea. A total of 108 haplotypes were observed from 73 variable sites. These domestic chicken mtDNA sequences could be assigned into seven clades (A-G). The results indicated that 71.97% of the black-bone haplotypes were related to the reference sequence that may originate from Eurasia, while the minor part of mtDNA sequences presumably derive from Southeast Asia, China, and Japan. Three clades were shared by Korean, Japanese, and Chinese black-bone chickens. These results provide basic data useful for making new breeding and conservation strategies for the black-bone chicken in China.

Association between -1082G/A, -819C/T, and -592C/A genetic polymorphisms in IL-10 and risk of type 2 diabetes mellitus in a Chinese population.

Type 2 diabetes mellitus is the most common form of endocrine disease in humans; genetic factors are known to contribute to the development of this disease. In this case-control study, we investigated the relationship between the -1082G/A, -819C/T, and -592C/A polymorphisms in interleukin 10 (IL-10) and the pathogenesis of type 2 diabetes mellitus in a Chinese population. Patients with type 2 diabetes mellitus (N = 228) and control subjects (N = 240) were recruited from the Department of Endocrinology at the People's Hospital of Linyi City, between September 2013 and April 2015. The IL-10 -1082G/A, -819C/T, and -592C/A polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism. Multivariate logistic regression analyses revealed that patients carrying the AA genotype of IL-10 -592C/A were at a higher risk of developing type 2 diabetes mellitus compared to those carrying the CC genotype [adjusted odds ratio (OR) = 1.74; 95% confidence interval (CI) = 1.03-2.95]. In addition, individuals carrying the A allele of IL-10 -592C/A showed a 1.34-fold higher risk of developing type 2 diabetes mellitus compared to those carrying the C allele (adjusted OR = 1.34; 95%CI = 1.03- 1.75). There was no significant correlation between the IL-10 -1082G/ A and -819C/T polymorphisms and risk of type 2 diabetes mellitus. In conclusion, this study shows that the -1082G/A polymorphism of IL-10 contributes to the onset of type 2 diabetes mellitus, and may be considered a biomarker for early screening of type 2 diabetes mellitus in the Chinese population studied here.

Effect of MSTN propeptide protein on the growth and development of Altay lamb muscle.

Prokaryotic expression technology was used to express maltose-binding protein binding myostatin (MSTN) propeptide fusion protein. Six disease-free Altay lambs were used in this study. The right leg gastrocnemii were injected with MSTN recombinant propeptide protein. The left leg gastrocnemii (the control group) were injected with the same dose of phosphate based saline. The lambs were fed during four months under the same conditions and then slaughtered. Gastrocnemius samples were hematoxylin-eosin stained and the size of the muscle fibers was measured. A real-time polymerase chain reaction (RT-PCR) showed that single gastrocnemius cells in the experimental group had an average area of 1163.01 µm(2), while it was 845.09 µm(2) in the control group (P < 0.05). This indicates that the MSTN propeptide biological agents had an inhibitory effect on MSTN. In order to reveal its mechanism, RT-PCR was conducted to detect the expression of the differentiation-associated genes MyoD, Myf5, Myogenin, p21, and Smad3. The results showed that, in the MSTN propeptide biological agent injected group, expression levels of MSTN, Smad3, and p21 were lower than the control group, while Myf5, MyoD, and Myogenin were higher compared to the control group. This indicates that, when expression of the MSTN gene was inhibited, muscle cell differentiation and growth can be promoted by Smad3 up-regulated expression of Myf5, MyoD, and Myogenin.

Sequence analysis of peste des petits ruminants virus from ibexes in Xinjiang, China.

Peste des petits ruminants (PPR) is an infectious disease caused by peste des petits ruminants virus (PPRV). While PPR mainly affects domestic goats and sheep, it also affects wild ungulates such as ibex, blue sheep, and gazelle, although there are few reports regarding PPRV infection in wild animals. Between January 2015 and February 2015, it was found for the first time that wild ibexes died from PPRV infection in Bazhou, Xinjiang, China, where a total of 38 ibexes (including young and adult ibexes) were found to have died abnormally from PPR-related issues. First, we tested for the presence of the F gene of PPRV by RT-PCR. Then, we compared the sequence of the isolated F gene from the ibex strain, termed PPRV Xinjiang/Ibex/2015, with those previously identified from small domestic ruminants from local areas near where the reported isolate was collected as well as those from other regions. The current sequence was phylogenetically classified as a lineage IV virus, and shared a high level of sequence identity (99.7%) with a previously described Xinjiang PPRV isolate.

Mechanism of SEMA3B gene silencing and clinical significance in glioma.

The aim of the current study was to explore mechanisms of SEMA3B gene expression and its clinical significance in glioma, and provide a theoretical foundation for investigating individualized treatment in glioma. Paraffin-embedded tissues from 43 patients with a confirmed clinical diagnosis of glioma following neurosurgery at the First Affiliated Hospital of Zhengzhou University from December 2013 to April 2014 were selected randomly. An additional three normal brain tissues were obtained following encephalic decompression excision due to acute craniocerebral injury in the same period, which were used as the control group. Immunohistochemical staining for vascular endothelial growth factor was performed on the glioma tissues from the 43 patients. Genomic DNA was extracted for bisulfate conversion and sequencing. SEMA3B was fully expressed in the three normal brain tissues, and incompletely expressed in the 43 glioma tissues, with a lack of expression in 48.8% (21/43) of samples. Moreover, 58% of high-grade gliomas (grade III and IV) lacked SEMA3B expression, which was significantly more than those that lacked expression (20%) in low-grade gliomas (grade I and II), indicating that, as the clinical pathological grade increased, SEMA3B expression decreased. The occurrence and development of malignant tumors is a product of multiple genes and other factors. Here, we provide theoretical basis for glioma development and prognosis involving DNA-methylation driven silencing of SEMA3B, and thus, SEMA3B is a potential target for directed treatments against glioma.

ATAD2 is overexpressed in gastric cancer and serves as an independent poor prognostic biomarker.

OBJECTIVE: ATPase family, AAA domain containing 2 (ATAD2) has been found overexpressed in various cancer types and correlated with malignant status and poor prognosis. However, little is known about the clinical significance of ATAD2 in gastric cancer patients. The aim of this study was to explore the clinical and prognostic significance of ATAD2 in gastric cancer. METHODS: The mRNA and protein levels expression of ATAD2 were detected in clinical tissue samples by qRT-PCR and immunohistochemistry, respectively. We examined the ATAD2 protein expression by immunohistochemistry. Furthermore, we analyzed the association between ATAD2 expression and clinicopathological features including prognosis in 166 gastric cancer samples. RESULTS: In our results, ATAD2 mRNA and protein were highly expressed in gastric cancer samples. ATAD2 overexpression was correlated with advanced clinical stage, tumor depth, lymph node metastasis, and distant metastasis. According to the survival analysis, ATAD2 protein overexpression was a poor independent prognostic factor for gastric cancer patients. CONCLUSIONS: In summary, ATAD2 could serve as a prognostic biomarker for gastric cancer patients.

Distribution of mitochondrial DNA fragments in the nuclear genome of the honeybee.

Nuclear mitochondrial pseudogenes (numts), which originated from mitochondrial DNA (mtDNA) insertions in the nuclear genome, have been detected in many species. The distribution of numts in the honeybee nuclear genome has not yet been fully reported. By referring to the whole honeybee mtDNA sequence and to the recent version of the honeybee nuclear genome, 236 reference sequences were identified by BLAST, with 90 unmapped. The size of the numts ranged from 219 to 3788 bp, and the homologous identity between numts and their corresponding mtDNA fragments varied from 71 to 93%. Furthermore, identified honeybee numts covered nearly all mitochondrial genes and were distributed over all chromosomes. This study provides useful information for further research related to mitochondrial genes and the evolution of the honeybee.

Male-specific association of the APC rs383830 T allele with the risk of coronary heart disease.

APC is a tumor suppressor gene that is involved in the processes of cell migration and adhesion, transcriptional activation, and apoptosis. The goal of this study was to evaluate the contribution of the APC rs383830 polymorphism to coronary heart disease (CHD) in Han Chinese. A total of 783 patients with CHD and 737 controls were tested in the current association study. Although our study did not identify an association between the APC rs383830 polymorphism and CHD, a breakdown analysis by gender indicated there was a significant contribution of the rs383830 T allele to the risk of CHD in males (P = 0.046, odds ratio = 1.267, 95% confidence interval = 1.004-1.598). In conclusion, our study suggested a male-specific association of the APC rs383830 polymorphism with CHD.

Expression of recombinant myostatin propeptide pPIC9K-Msp plasmid in Pichia pastoris.

Myostatin propeptide can inhibit the biological activity of myostatin protein and promote muscle growth. To express myostatin propeptide in vitro with a higher biological activity, we performed codon optimization on the sheep myostatin propeptide gene sequence, and mutated aspartic acid-76 to alanine based on the codon usage bias of Pichia pastoris and the enhanced biological activity of myostatin propeptide mutant. Modified myostatin propeptide gene was cloned into the pPIC9K plasmid to form the recombinant plasmid pPIC9K-Msp. Recombinant plasmid pPIC9K-Msp was transformed into Pichia pastoris GS115 by electrotransformation. Transformed cells were screened, and methanol was used to induce expression. SDS-PAGE and western blotting were used to verify the successful expression of myostatin propeptide with biological activity in Pichia pastoris, providing the basis for characterization of this protein.

Molecular cytogenetic identification of a novel 1AL.1RS translocation line with powdery mildew resistance.

A wheat germplasm line 13-2-2 with resistance to powdery mildew was isolated; this line was derived from common wheat cv. W770B and rye, Secale cereale L. (2n = 2x = 14, RR). The line was characterized based on cytological, genomic in situ hybridization (GISH), sequence-characterized amplified region (SCAR), and simple sequence repeat (SSR) analyses. The mitotic and meiotic investigations showed that the chromosome number and configuration of 13-2-2 were 2n = 42 = 21 II. GISH using rye genomic DNA as a probe detected a pair of R genome chromosome arms with strong hybridization signals in 13-2-2. Three 1RS chromosome-specific SCAR markers amplified 1RS specific bands in line 13-2-2. We screened 320 SSR primer pairs on the long or short arms from seven wheat homoeologous groups in the translocation line and parents. However, only three 1AS primers could not be amplified in line 13-2-2, whereas the others were amplified. Thus, these markers suggested that the line 13-2-2 was 1AL.1RS translocation line. Line 13-2-2 was immune to powdery mildew after inoculation with Blumeria graminis f. sp. tritici isolates E05 and E07 during the adult plant stages. In contrast, the maternal parent W770B, Kavkaz with Pm8, and Amigo with Pm17 were heavily infected with spores and had reaction response scores of susceptible. Thus, the new wheat-rye 1AL.1RS translocation line with resistance to powdery mildew could be a new and valuable donor source for wheat improvement. The molecular markers developed in this study might also be useful tools for marker-assisted selection.

Cytogenetic and molecular identification of a wheat-Leymus mollis alien multiple substitution line from octoploid Tritileymus x Triticum durum.

Leymus mollis (Trin.) Pilger (NsNsXmXm, 2n = 28), a wild relative of common wheat, possesses many traits that are potentially valuable for wheat improvement. In order to exploit and utilize the useful genes of L. mollis, we developed a multiple alien substitution line, 10DM50, from the progenies of octoploid Tritileymus M842-16 x Triticum durum cv. D4286. Genomic in situ hybridization analysis of mitosis and meiosis (metaphase I), using labeled total DNA of Psathyrostachys huashanica as probe, showed that the substitution line 10DM50 was a cytogenetically stable alien substitution line with 36 chromosomes from wheat and three pairs of Ns genome chromosomes from L. mollis. Simple sequence repeat analysis showed that the chromosomes 3D, 6D, and 7D were absent in 10DM50. Expressed sequence tag-sequence tagged sites analysis showed that new chromatin from 3Ns, 6Ns, and 7Ns of L. mollis were detected in 10DM50. We deduced that the substitution line 10DM50 was a multiple alien substitution line with the 3D, 6D, and 7D chromosomes replaced by 3Ns, 6Ns, and 7Ns from L. mollis. 10DM50 showed high resistance to leaf rust and significantly improved spike length, spikes per plant, and kernels per spike, which are correlated with higher wheat yield. These results suggest that line 10DM50 could be used as intermediate material for transferring desirable traits from L. mollis into common wheat in breeding programs.

The significant increase and dynamic changes of the myeloid-derived suppressor cells percentage with chemotherapy in advanced NSCLC patients.

PURPOSE: To investigate the correlations between myeloid-derived suppressor cells (MDSCs) in the peripheral blood and cancer stage, immune function, and chemotherapy. METHODS: Percentages of MDSCs (CD11b(+)CD14(-)CD33(+) cells) and lymphocyte subsets in peripheral blood mononuclear cells (PBMCs) of 94 patients with Non-small cell lung cancer (NSCLC) who were treated naïve and 30 healthy individuals were measured. Changes of the MDSCs percentage were further detected in patients with advanced NSCLC treated with systemic chemotherapy. Finally, coculture with CD8(+) cells was developed to determine effect of MDSCs on IFN-γ secretion of T lymphocytes. RESULTS: MDSCs percentage of 94 patients with NSCLC was significantly higher than that of 30 healthy subjects (P < 0.05), the percentages were increased with tumor progression, in patients with stage III and IV percentages were significantly higher than those in stage I and II patients (P = 0.013). The MDSCs percentage was negatively related to percentage of CD8(+) cells in the peripheral blood (r = -0.354, n = 38, P = 0.029), and when they were cocultured, IFN-γ secretion of CD8(+) cells was significantly decreased (P < 0.05). In 20 patients with advanced NSCLC who received systemic chemotherapy, nine partial remission (PR) cases got MDSCs percentage significantly decreased (P < 0.001), three stable disease (SD) cases remained invariable (P = 0.307) and eight progressive disease (PD) cases got significantly increased (P = 0.024). CONCLUSION: The percentage of MDSCs in the patients was significantly higher than that of the healthy control subjects and it increased with tumor progression partially by inhibiting the CD8(+) cell function. The dynamic changes of MDSCs percentage reflected the efficacy of systemic chemotherapy.

A novel SCAR marker for detecting Psathyrostachys huashanica Keng chromatin introduced in wheat.

In this study, we cloned and sequenced a 938-base pair polymorphic band, pHs27, in the tightly linked random amplified polymorphic DNA marker OPU10 and converted it into a sequence-characterized amplified region (SCAR) marker referred to as RHS141, which was specific for the Ns genome of Psathyrostachys huashanica. A GenBank basic local alignment search tool search showed that the sequence of pHs27 had no primary sequence homology with known sequences, and Southern blotting confirmed this result. This SCAR marker was used to detect Ns genome chromatin in wheat, and it was successfully amplified in P. huashanica itself, a complete set of wheat-P. huashanica disomic addition lines (1Ns-7Ns), and undetermined homoeologous group addition lines. This SCAR marker will be a powerful tool for the marker-assisted selection of P. huashanica chromosome(s) in a wheat background, and it should also allow wheat breeders to screen for the excellent traits found in P. huashanica chromatin.

Testing the utility of mitochondrial cytochrome oxidase subunit 1 sequences for phylogenetic estimates of relationships between crane (Grus) species.

Morphology and biogeography are widely used in animal taxonomy. Recent study has suggested that a DNA-based identification system, using a 648-bp portion of the mitochondrial gene cytochrome oxidase subunit 1 (CO1), also known as the barcoding gene, can aid in the resolution of inferences concerning phylogenetic relationships and for identification of species. However, the effectiveness of DNA barcoding for identifying crane species is unknown. We amplified and sequenced 894-bp DNA fragments of CO1 from Grus japonensis (Japanese crane), G. grus (Eurasian crane), G. monacha (hooded crane), G. canadensis (sandhill crane), G. leucogeranus (Siberian crane), and Balearica pavonina (crowned crane), along with those of 15 species obtained from GenBank and DNA barcoding, to construct four algorithms using Tringa stagnatilis, Scolopax rusticola, and T. erythropus as outgroups. The four phylum profiles showed good resolution of the major taxonomic groups. We concluded that reconstruction of the molecular phylogenetic tree can be helpful for classification and that CO1 sequences are suitable for studying the molecular evolution of cranes. Although support for several deeper branches was limited, CO1 data gave remarkably good separations, especially considering that our analysis was based on just a fragment of the gene and that CO1 has generally been viewed as useful only for resolving shallow divergences.