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Abstract Climatic factors play an essential role in the growth of tree ring width. In this study, we aimed to evaluate the correlation between climatic variables and tree-ring growth characteristics of Pinus sibirica in Altai mountains, northwestern China. This study being is first of its kind on climate growth analysis of Pinus sibirica in northwestern China. The study showed great potential to understand the species growing under the specific climatic conditions. Total of 70 tree cores collected from three sites in the sampling area, out of which 63 tree cores considered for this study. The effect of climatic variables which was studied include precipitation, temperature and PDSI. Our results showed that Tree Ring Width chronology has a significantly positive correlation with the late winter (March) temperature and significant negative correlation with the July temperatures. A significant correlation was observed with the late summer precipitation whereas no significant relation found with the Palmer Drought Severity Index. These significant correlations with temperature and precipitation suggested that this tree species had the potential for the reconstruction of the past climate in the area.

RESUMO Fatores climáticos desempenham papel essencial no crescimento da largura dos anéis das árvores. Neste estudo, objetivou-se avaliar a correlação entre variáveis climáticas e características de crescimento de anéis de árvores de Pinus sibirica nas montanhas de Altai, noroeste da China. Este estudo é o primeiro desse tipo na análise do crescimento climático de Pinus sibirica no noroeste da China. O estudo mostrou grande potencial para entender as espécies que crescem sob as condições climáticas específicas. Total de 70 testemunhos coletados em três locais na área de amostragem, dos quais 63 testemunhos considerados para este estudo. O efeito das variáveis climáticas estudadas incluem precipitação, temperatura e PDSI. Nossos resultados mostraram que a cronologia da Largura do Anel da Árvore tem uma correlação significativamente positiva com a temperatura do final do inverno (março) e uma correlação negativa significativa com as temperaturas de julho. Uma correlação significativa foi observada com a precipitação do final do verão, enquanto nenhuma relação significativa foi encontrada com o Índice de Severidade de Seca de Palmer. Essas correlações significativas com a temperatura e precipitação sugeriram que esta espécie de árvore tinha o potencial para a reconstrução do clima passado na área.

RESUMO

Climatic factors play an essential role in the growth of tree ring width. In this study, we aimed to evaluate the correlation between climatic variables and tree-ring growth characteristics of Pinus sibirica in Altai mountains, northwestern China. This study being is first of its kind on climate growth analysis of Pinus sibirica in northwestern China. The study showed great potential to understand the species growing under the specific climatic conditions. Total of 70 tree cores collected from three sites in the sampling area, out of which 63 tree cores considered for this study. The effect of climatic variables which was studied include precipitation, temperature and PDSI. Our results showed that Tree Ring Width chronology has a significantly positive correlation with the late winter (March) temperature and significant negative correlation with the July temperatures. A significant correlation was observed with the late summer precipitation whereas no significant relation found with the Palmer Drought Severity Index. These significant correlations with temperature and precipitation suggested that this tree species had the potential for the reconstruction of the past climate in the area.(AU)

Fatores climáticos desempenham papel essencial no crescimento da largura dos anéis das árvores. Neste estudo, objetivou-se avaliar a correlação entre variáveis climáticas e características de crescimento de anéis de árvores de Pinus sibirica nas montanhas de Altai, noroeste da China. Este estudo é o primeiro desse tipo na análise do crescimento climático de Pinus sibirica no noroeste da China. O estudo mostrou grande potencial para entender as espécies que crescem sob as condições climáticas específicas. Total de 70 testemunhos coletados em três locais na área de amostragem, dos quais 63 testemunhos considerados para este estudo. O efeito das variáveis climáticas estudadas incluem precipitação, temperatura e PDSI. Nossos resultados mostraram que a cronologia da Largura do Anel da Árvore tem uma correlação significativamente positiva com a temperatura do final do inverno (março) e uma correlação negativa significativa com as temperaturas de julho. Uma correlação significativa foi observada com a precipitação do final do verão, enquanto nenhuma relação significativa foi encontrada com o Índice de Severidade de Seca de Palmer. Essas correlações significativas com a temperatura e precipitação sugeriram que esta espécie de árvore tinha o potencial para a reconstrução do clima passado na área.(AU)

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RESUMO

Climatic factors play an essential role in the growth of tree ring width. In this study, we aimed to evaluate the correlation between climatic variables and tree-ring growth characteristics of Pinus sibirica in Altai mountains, northwestern China. This study being is first of its kind on climate growth analysis of Pinus sibirica in northwestern China. The study showed great potential to understand the species growing under the specific climatic conditions. Total of 70 tree cores collected from three sites in the sampling area, out of which 63 tree cores considered for this study. The effect of climatic variables which was studied include precipitation, temperature and PDSI. Our results showed that Tree Ring Width chronology has a significantly positive correlation with the late winter (March) temperature and significant negative correlation with the July temperatures. A significant correlation was observed with the late summer precipitation whereas no significant relation found with the Palmer Drought Severity Index. These significant correlations with temperature and precipitation suggested that this tree species had the potential for the reconstruction of the past climate in the area.

Fatores climáticos desempenham papel essencial no crescimento da largura dos anéis das árvores. Neste estudo, objetivou-se avaliar a correlação entre variáveis climáticas e características de crescimento de anéis de árvores de Pinus sibirica nas montanhas de Altai, noroeste da China. Este estudo é o primeiro desse tipo na análise do crescimento climático de Pinus sibirica no noroeste da China. O estudo mostrou grande potencial para entender as espécies que crescem sob as condições climáticas específicas. Total de 70 testemunhos coletados em três locais na área de amostragem, dos quais 63 testemunhos considerados para este estudo. O efeito das variáveis climáticas estudadas incluem precipitação, temperatura e PDSI. Nossos resultados mostraram que a cronologia da Largura do Anel da Árvore tem uma correlação significativamente positiva com a temperatura do final do inverno (março) e uma correlação negativa significativa com as temperaturas de julho. Uma correlação significativa foi observada com a precipitação do final do verão, enquanto nenhuma relação significativa foi encontrada com o Índice de Severidade de Seca de Palmer. Essas correlações significativas com a temperatura e precipitação sugeriram que esta espécie de árvore tinha o potencial para a reconstrução do clima passado na área.

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The current experiment aimed to evaluate the effects of Pancreatin supplementation at different levels on ileal and fecal digestibility in layer-type cockerels. A total of 480-day-old silver brown Hy-Line male chicks were randomly allocated into 5 treatments, 6 replicates (16 birds per pen) arranged in a completely randomized design. Pancreatin enzyme was supplemented on a basal corn-soybean meal-based diet at 0, 250, 500, 750, and 1000 mg/kg and was fed in two growth phases (starter and grower). The results indicated that at the end of the starter stage, except for 1000 mg/kg, dietary Pancreatin supplementation levels increased (p<0.05) the ileal crude protein (CP). Similarly, addition of Pancreatin increased (p< 0.05) apparent ileal amino acids (AA) digestibility (AIAAD) total means of AA (MTAA), means of indispensable AA (MIAA) and dispensable AA (MDAA) with the optimum performance on 250 mg/kg and 500 mg/kg. However, except for histidine and alanine which were negatively affected (p<0.05), and MIAA, MDAA, MTAA which were also positively affected, the addition level at 1000 mg/kg did not affect most of the AIAAD compared to the non-supplemented. Further, Pancreatin supplementation had no effect (p>0.05) on nitrogen digestibility (ND), nitrogen retention (NR), digestible energy (DE), apparent metabolizable energy (AME), dry matter digestibility (DMD), dry matter retention (DMR), and apparent metabolizable energy corrected for nitrogen (AME-n) on fed starter diet. On the other hand, at the end of the grower stage, dietary Pancreatin enzyme supplementation reduced (p<0.05) the ileal CP, MIAA, MDAA, MTAA, AIAAD, AME, AME-n, ND, NR, DE, DMD, and DMR in a dose-dependent manner. The rate of reduction was more marked on Pancreatin addition level 1000 mg/kg. In conclusion, Pancreatin supplementation at 250 mg/kg, 500 mg/kg, and 750 mg/kg improved AIAAD and ileal CP, especially at the young age. The rate of pancreatin enzyme effect was dependent on enzyme supplement level to the ileal CP and individual amino acid.(AU)

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Climatic factors play an essential role in the growth of tree ring width. In this study, we aimed to evaluate the correlation between climatic variables and tree-ring growth characteristics of Pinus sibirica in Altai mountains, northwestern China. This study being is first of its kind on climate growth analysis of Pinus sibirica in northwestern China. The study showed great potential to understand the species growing under the specific climatic conditions. Total of 70 tree cores collected from three sites in the sampling area, out of which 63 tree cores considered for this study. The effect of climatic variables which was studied include precipitation, temperature and PDSI. Our results showed that Tree Ring Width chronology has a significantly positive correlation with the late winter (March) temperature and significant negative correlation with the July temperatures. A significant correlation was observed with the late summer precipitation whereas no significant relation found with the Palmer Drought Severity Index. These significant correlations with temperature and precipitation suggested that this tree species had the potential for the reconstruction of the past climate in the area.

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OBJECTIVES: Although studies have reported that miR-596 extensively participates in multiple cancer progression, the biological mechanisms and effects of miR-596 in prostatic cancer remain unclear. The literature is aimed to reveal the function and possible molecular mechanisms of miR-596 in prostatic cancer carcinogenesis. MATERIALS AND METHODS: qRT-PCR was applied to examine miR-596 expression in prostatic cancer cell lines and samples, also methylation-specific PCR was used to detect the methylation status of the promoter CpG islands in prostatic cancer samples. Meanwhile, the tumor-related effects of miR-596 were detected via cell viability, clone formation assay, migration assay, flow cytometric and AO/EB assay. qRT-PCR and Western blots were applied to investigate the function of miR-596 on malignant behavior in prostatic cancer cells. RESULTS: We found that miR-596 mRNA was decreased in prostatic cancer samples and cell lines. miR-596 mRNA level was also correlated to cancer stage, Gleason scores, while miR-596 promoter methylation was related to cancer tumor stage, Gleason score and preoperative PSA levels. miR-596 inhibited the cell growth and activity by causing cell apoptosis, and also suppressed the migration of prostatic cancer cells by revealing the epithelial-mesenchymal transition process. In addition, Western blot indicates that miR-596 overexpression deregulated Wnt/ß-catenin signaling, by restraining phosphorylation levels of ß-catenin and expression levels of downstream targets. CONCLUSIONS: In summary, this research indicates that miR-596 overexpression could be potentially useful in the cell growth and migration of prostatic cancer and serves as a potential molecular marker in prostatic cancer.

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ABSTRACT The present study was conducted to investigate the effects of dietary fiber on growth performance, fat deposition, serum lipids, fat metabolism, and mRNA (messenger RNA) expression of lipoprotein lipase (LPL) in Jilin white and Carlos geese. Sixty Jilin white and sixty Carlos geese aged six-weeks and of similar health and weight (average weight 313.11g) were selected. Geese of each breed were randomly divided into two groups (n=30), and with each group containing three replicate subgroups of 10 geese. The diet was supplemented with 8% or 11% fiber (corn straw powder). The Jilin white geese are divided into A1 (8%) and A2 (11%) groups, and Carlos geese are divided into B1 (8%) and B2 (11%) groups. The experiment lasted 35 days. The results showed that high dietary fiber can significantly (p 0.05) increase average daily feed intake (ADFI), significantly (p 0.05) reduce final weight (FW) and average daily gain (ADG) of both varieties, and increase LPL mRNA expression levels in abdominal fat, liver, sebum, and urethral glands. High dietary fiber accelerates intestinal peristalsis, affects the absorption of other nutrients, reduces the available energy value of the absorbed feed, and increases fat loss. Compared with the to Carlos geese, high dietary fiber content had a more significant effect on the live, slaughter, and sebum weights and sebum percentage of the Jilin white geese, indicating that the Carlos geese have higher requirements for dietary fiber content. High fiber content will reduce the growth performance, slaughter performance, and fat deposition of geese.

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Circulating tumor cells (CTCs) are cells that have shed into the vasculature or lymphatics from a primary tumor and are carried around the body in the blood circulation. CTCs undergo a series of migration, adhesion and aggregation to form metastases, leading to post-operative recurrence and metastasis in patients with malignant tumors. The detection and analysis of CTCs, as a new non-invasive diagnostic tool, plays an important role in tumor diagnosis, therapeutic efficacy, monitoring recurrence, prognosis assessment and tumor precision medical treatment.

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The objectives of this study were to investigate the effects of tributyrin (TB) and coated sodium butyrate (CSB) on intestinal morphology, disaccharidase activity and intramuscular fat of broilers challenged with lipopolysaccharide (LPS). A total of 160 1-day-old healthy Cobb broilers were randomly allocated into four groups: (1) control; (2) LPS, in which broilers were fed a basal diet and intraperitoneally injected with 500 g/kg LPS on days 38, 40 and 42; (3) TB, in which LPS-challenged broilers were fed basal diet supplemented with 500 mg/kg TB; and (4) CSB, in which LPS-challenged broilers were fed basal diet supplemented with 877 mg/kg CSB. Addition of TB and CSB inhibited (p<0.05) the decrease in villus height in the duodenum and ileum of LPS-challenged broilers, respectively. Both TB and CSB increased (p<0.05) activity of maltase in the small intestine, and TB increased (p<0.05) activity of isomaltase in the ileum. Additionally, dietary addition of TB and CSB decreased (p<0.05) the content of intramuscular fat. In conclusion, dietary supplementation of TB was more effective than CSB in improving intestinal morphology and disaccharidase activity of LPS-challenged broilers, and they both reduced intramuscular fat in the breast and legs.

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The objectives of this study were to investigate the effects of tributyrin (TB) and coated sodium butyrate (CSB) on intestinal morphology, disaccharidase activity and intramuscular fat of broilers challenged with lipopolysaccharide (LPS). A total of 160 1-day-old healthy Cobb broilers were randomly allocated into four groups: (1) control; (2) LPS, in which broilers were fed a basal diet and intraperitoneally injected with 500 g/kg LPS on days 38, 40 and 42; (3) TB, in which LPS-challenged broilers were fed basal diet supplemented with 500 mg/kg TB; and (4) CSB, in which LPS-challenged broilers were fed basal diet supplemented with 877 mg/kg CSB. Addition of TB and CSB inhibited (p<0.05) the decrease in villus height in the duodenum and ileum of LPS-challenged broilers, respectively. Both TB and CSB increased (p<0.05) activity of maltase in the small intestine, and TB increased (p<0.05) activity of isomaltase in the ileum. Additionally, dietary addition of TB and CSB decreased (p<0.05) the content of intramuscular fat. In conclusion, dietary supplementation of TB was more effective than CSB in improving intestinal morphology and disaccharidase activity of LPS-challenged broilers, and they both reduced intramuscular fat in the breast and legs.(AU)

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Ovarian cancer is one of the most common malignancies in women. Semaphorin 4D (sema 4D) is involved in the progress of multiple cancers. In the presence of estrogen-like ligands, estrogen receptors (ERα and ERß) participate in the progress of breast and ovarian cancers by transcriptional regulation. The aim of the study was to investigate the role of sema 4D and elucidate the regulatory pattern of ERα and ERß on sema 4D expression in ovarian cancers. Sema 4D levels were up-regulated in ovarian cancer SKOV-3 cells. Patients with malignant ovarian cancers had significantly higher sema 4D levels than controls, suggesting an oncogene role of sema 4D in ovarian cancer. ERα expressions were up-regulated in SKOV-3 cells compared with normal ovarian IOSE80 epithelial cells. Conversely, down-regulation of ERß was observed in SKOV-3 cells. Forced over-expression of ERα and ERß in SKOV-3 cells was manipulated to establish ERα+ and ERß+ SKOV-3 cell lines. Incubation of ERα+ SKOV-3 cells with ERs agonist 17ß-estradiol (E2) significantly enhanced sema 4D expression and rate of cell proliferation. Incubated with E2, ERß+ SKOV-3 cells showed lower sema 4D expression and cell proliferation. Blocking ERα and ERß activities with ICI182-780 inhibitor, sema 4D expressions and cell proliferation of ERα+ and ERß+ SKOV-3 cells were recovered to control levels. Taken together, the data showed that sema 4D expression was positively correlated with the progress of ovarian cancer. ERα positively regulated sema 4D expression and accelerated cell proliferation. ERß negatively regulated sema 4D expression and inhibited cell multiplication.

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PURPOSE: The prognostic factors for the survival of small cell lung cancer (SCLC) patients are still widely debated. The aim of this study was to identify the clinical features and prognostic factors in SCLC patients. METHODS: A retrospective study was conducted on SCLC patients who were treated in our hospital between July 2010 and July 2015. Comparison of overall survival (OS) was performed using the Kaplan-Meier method. Prognostic factors for OS were identified by multivariate Cox regression models. RESULTS: A total of 523 patients with complete data and ECOG 0-2 were enrolled in our study. A total of 383 patients (73.2%) were diagnosed with ES-SCLC (extensive-stage SCLC) and 140 patients (26.8%) were diagnosed with LS-SCLC (limited-stage SCLC). In all patients, early disease stage, good ECOG, normal neuron-specific enolase (NSE), thoracic radiotherapy, ≥4 cycles of chemotherapy, prophylactic cranial irradiation, good response to initial therapy were independent favorable prognostic factors for OS, along with gender, age, CEA and CA125. In LS-SCLC patients, normal NSE, normal CEA, good response to initial therapy and surgery were independent favorable prognostic factors for OS. In ES-SCLC patients, good ECOG, normal NSE, thoracic radiotherapy, ≥4 cycles of chemotherapy, prophylactic cranial irradiation and good response to initial therapy were independent favorable prognostic factors for OS. Remarkably, NSE and response to initial therapy were independent prognostic factors for OS in all SCLC patients, LS-SCLC patients and ES-SCLC patients. CONCLUSION: The normal NSE and good response to initial therapy predicted a better survival for SCLC patients, regardless of disease stage.

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Ovarian cancer is one of the most common malignancies in women. Semaphorin 4D (sema 4D) is involved in the progress of multiple cancers. In the presence of estrogen-like ligands, estrogen receptors (ERα and ERβ) participate in the progress of breast and ovarian cancers by transcriptional regulation. The aim of the study was to investigate the role of sema 4D and elucidate the regulatory pattern of ERα and ERβ on sema 4D expression in ovarian cancers. Sema 4D levels were up-regulated in ovarian cancer SKOV-3 cells. Patients with malignant ovarian cancers had significantly higher sema 4D levels than controls, suggesting an oncogene role of sema 4D in ovarian cancer. ERα expressions were up-regulated in SKOV-3 cells compared with normal ovarian IOSE80 epithelial cells. Conversely, down-regulation of ERβ was observed in SKOV-3 cells. Forced over-expression of ERα and ERβ in SKOV-3 cells was manipulated to establish ERα+ and ERβ+ SKOV-3 cell lines. Incubation of ERα+ SKOV-3 cells with ERs agonist 17β-estradiol (E2) significantly enhanced sema 4D expression and rate of cell proliferation. Incubated with E2, ERβ+ SKOV-3 cells showed lower sema 4D expression and cell proliferation. Blocking ERα and ERβ activities with ICI182-780 inhibitor, sema 4D expressions and cell proliferation of ERα+ and ERβ+ SKOV-3 cells were recovered to control levels. Taken together, the data showed that sema 4D expression was positively correlated with the progress of ovarian cancer. ERα positively regulated sema 4D expression and accelerated cell proliferation. ERβ negatively regulated sema 4D expression and inhibited cell multiplication.

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Mareks disease (MD), a lymphoproliferative disorder of chickens caused by the MD virus (MDV), is economically significant. The resistance/susceptibility to MD is controlled by host genetics. The host response to different virus strains varies. The pathogenicity of REV-LTR deleted GX0101LTR MDV has been previously reported. However, the precise molecular mechanism of the response of chickens to GX0101LTR remains unclear. The current study aimed at identifying the genes and pathways involved in the response to GX0101LTR virus infection in specific pathogen-free chicken embryo fibroblast cells using global transcriptome analysis. A total of 1,633 genes associated with GX0101LTR infection were identified. Functional analysis showed that the cytokine-cytokine receptor interaction plays an important role in the response to GX0101LTR infection.

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Mareks disease (MD), a lymphoproliferative disorder of chickens caused by the MD virus (MDV), is economically significant. The resistance/susceptibility to MD is controlled by host genetics. The host response to different virus strains varies. The pathogenicity of REV-LTR deleted GX0101LTR MDV has been previously reported. However, the precise molecular mechanism of the response of chickens to GX0101LTR remains unclear. The current study aimed at identifying the genes and pathways involved in the response to GX0101LTR virus infection in specific pathogen-free chicken embryo fibroblast cells using global transcriptome analysis. A total of 1,633 genes associated with GX0101LTR infection were identified. Functional analysis showed that the cytokine-cytokine receptor interaction plays an important role in the response to GX0101LTR infection.(AU)

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Simple sequence repeats (SSRs), one of the most powerful molecular markers, can be used for DNA fingerprinting, variety identification, genetic mapping, and marker-assisted selection. Using the pear's (Pyrus pyrifolia Nakai) 75,764 unigenes (55,676,271 bp) obtained by deep transcriptome sequencing, a total of 10,622 novel SSRs were identified in 9154 unigenes, accounting for 14.02% of all unigenes. The average length and distribution of these SSRs was about 16 bp and 5.24 kb, respectively. Dinucleotide repeat motifs were the main type, with a frequency of 55.87%, followed by trinucleotides (24.45%). There were 159 kinds of repeat motifs existing in the pear transcriptome. AG/CT was the most frequent motif, accounting for 49.64%. All 9154 SSR-containing unigenes were functionally annotated using Nr (NCBI non-redundant protein database), Nt (NCBI non-redundant nucleotide database), and the Swiss-Prot database, and were classified further by Gene Ontology and Clusters of Orthologous Groups. In addition, a total of 4300 primer pairs were designed from all SSR loci obtained. Of these, 40 primers were randomly selected for PCR amplification and polyacrylamide gel (PAGE) analysis. Among the 40 primer pairs, 31 were successfully separated via PAGE. These findings also confirm that mining SSRs using next-generating sequencing technologies is a fast, effective, and reliable approach.

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Previous studies have shown a close correlation between the generation of B cell autoantibodies and imbalances in T lymphocyte subpopulations and the occurrence of disease. In this study, we have analyzed the effects of abnormal expression of CD4+CD25+-regulatory T cells, T lymphocyte subpopulations, immunoglobulins, complement factors, inflammatory factors, and adhesion molecules in the peripheral blood on the occurrence and development of autoimmune disease. Eighty patients with autoimmune disease were randomly (equally) divided into active-stage and stable-stage disease groups (according to pre-defined criteria). Fifty healthy people were recruited to the control group. The above-mentioned indices were detected by flow cytometry, immunity transmission turbidity, and enzyme-linked immunosorbent assay. We observed an obvious decrease in the CD4+CD25+- regulatory T cell, CD4+ cell, CD4+/CD8+ cell, NK cell, C3, and C4 expression in all three groups; however, this decrease was statistically significant in the active-stage group (P < 0.05). Alternately, we observed a significant increase in the expression of CD8+ cells, immunoglobulin (Ig) A, IgG, IgM, tumor necrosis factor-α, interleukin (IL)-10, IL-17, interferon-g, intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and E-selectin expression in the active-stage group (P < 0.05). Therefore, inflammatory reactions and immune dysfunction occurs during the active-stage of autoimmune disease, and detection of the immune indices and inflammatory and adhesion factors could help evaluate the immune stage in these patients, providing an experimental basis for the determination of disease progression and clinical treatment.

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Whether callose deposition is the cause or result of ovule sterility in Medicago sativa remains controversial, because it is unclear when and where changes in callose deposition and dissolution occur during fertile and sterile embryo sac formation. Here, alfalfa spontaneous multi-pistil mutant (mp1) and wild-type plants were used to compare the dynamics of callose deposition during embryo sac formation using microscopy. The results showed that both mutant and wild-type plants experienced megasporogenesis and megagametogenesis, and there was no significant difference during megasporogenesis. In contrast to the wild-type plants, in which the mature embryo sac was observed after three continuous cycles of mitosis, functional megaspores of mutant plants developed abnormally after the second round of mitosis, leading to degeneration of synergid, central, and antipodal cells. Callose deposition in both mutant and wild-type plants was first observed in the walls of megasporocytes, and then in the megaspore tetrad walls. After meiosis, the callose wall began to degrade as the functional megaspore underwent mitosis, and almost no callose was observed in the mature embryo sac in wild-type plants. However, callose deposition was observed in mp1 plants around the synergid, and increased with the development of the embryo sac, and was mainly deposited at the micropylar end. Our results indicate that synergid, central, and antipodal cells, which are surrounded by callose, may degrade owing to lack of nutrition. Callose accumulation around the synergid and at the micropylar end may hinder signals required for the pollen tube to enter the embryo sac, leading to abortion.

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A full-length cDNA sequence coding ribosomal protein S3a of mulberry tree, which we designated MmRPS3a (GenBank accession No. KR610331), was cloned based on mulberry expressed sequence tags. Sequence analysis showed that the MmRPS3a is 1089 bp long and contains a 80-bp 5'-UTR (untranslated region) and a 220-bp 3'-UTR. Its open reading frame consists of a 789-bp encoding 262 amino acids with a predicted molecular weight of 30.053 kDa and an isoelectric point of 9.84. Homology analysis revealed that MmRPS3a gene is highly conservative in mulberry and other species including Morus notabilis, Theobroma cacao, and Ricinus communis. Phylogenetic analysis based on MmRPS3a of other species showed that mulberry had a closer relationship with Prunus persica, Arabidopsis thaliana, Solanum tuberosum, Solanum lycopersicum, and Vitis vinifera. The results of quantitative PCR analysis showed that the transcriptional level of MmRPS3a mRNA changed significantly under the conditions of hypothermia, aridity, salt stress, and varieties of differing resistances.

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RT-PCR was used to study the temporal and spatial pattern of Yes-associated protein 1 (YAP1) and myosin heavy chain (MyHC) expression in four different skeletal muscles (i.e., longissimus dorsi muscle, soleus muscle, gastrocnemius muscle, and extensor digitorum longus) and three growth stages (i.e., 2 days old, 2 and 6 months old) of Hu Sheep. The results showed that YAP1 was differentially expressed in skeletal muscles of sheep, that expression increased gradually with age, and that there were high levels of expression in the gastrocnemius muscle and lower levels in the longissimus dorsi muscle. MyHCI was expressed at high levels in the soleus muscle and at lower levels in the longissimus dorsi muscle. In contrast, MyHCIIA and MyHCIIX were expressed at high levels in the extensor digitorum longus and at lower levels in the soleus muscle. The expression of MyHCI and MyHCIIA decreased with increasing age while that of MyHCIIX increased. YAP1 expression was negatively correlated with MyHCII (P < 0.01) and positively correlated with MyHCIIX (P < 0.01) across all growth stages and skeletal muscle types studied. We speculate that after birth, the thicker muscle fiber diameter is associated with the high expression of MyHCIIX. Therefore, we conclude that YAP1 expression affects sheep muscle fiber development after birth and provides important genetic information for the selection candidate genes for sheep muscle growth.

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