Hepatopancreas necrosis disease (HPND) of the Chinese mitten crab Eriocheir sinensis causes huge economic loss in China. However, the pathogenic factors and pathogenesis are still a matter of dissension. To search for potential pathogens, the hepatopancreatic flora of diseased crabs with mild symptoms, diseased crabs with severe symptoms, and crabs without visible symptoms were investigated using metatranscriptomics sequencing. The prevalence of Absidia glauca and Candidatus Synechococcus spongiarum decreased, whereas the prevalence of Spiroplasma eriocheiris increased in the hepatopancreatic flora of crabs with HPND. Homologous sequences of 34 viral species and 4 Microsporidian species were found in the crab hepatopancreas without any significant differences between crabs with and without HPND. Moreover, DEGs in the hepatopancreatic flora between crabs with severe symptoms and without visible symptoms were enriched in the ribosome, retinol metabolism, metabolism of xenobiotics by cytochrome P450, drug metabolism—cytochrome P450, biosynthesis of unsaturated fatty acids, and other glycan degradation. Moreover, the relative abundance of functions of DEDs in the hepatopancreatic flora changed with the pathogenesis process. These results suggested that imbalance of hepatopancreatic flora was associated with crab HPND. The identified DEGs were perhaps involved in the pathological mechanism of HPND; nonetheless, HPND did not occur due to virus or microsporidia infection.

Encapsulation of antigens within protein microcrystals (polyhedra) is a promising approach for the stable delivery of vaccines. In this study, a vaccine was encapsulated into polyhedra against cyprinid herpesvirus II (CyHV-2). CyHV-2 typically infects gibel carp, Carassius auratus gibelio, causing gill hemorrhagic disease. The vaccine was constructed using a codon-optimized sequence, D4ORF, comprising the ORF72 (region 1–186 nt), ORF66 (region 993–1197 nt), ORF81 (region 603–783 nt), and ORF82 (region 85–186 nt) genes of CyHV-2. The H1-D4ORF and D4ORF-VP3 sequences were, respectively, obtained by fusing the H1-helix sequence (region 1–90 nt) ofBombyx mori cypovirus(BmCPV) polyhedrin to the 5′ terminal end of D4ORF and by fusing a partial sequence (1–279 nt) of the BmCPV VP3 gene to the 3′ terminal end of D4ORF. Furthermore, BmNPV-H1-D4ORF-polh and BmNPV-D4ORF-VP3-polh recombinant B. mori nucleopolyhedroviruses (BmNPVs), belonging to the family Baculoviridae, and co-expressing BmCPV polyhedrin and H1-D4ORF or D4ORF-VP3, were constructed. H1-D4ORF and D4ORF-VP3 fusion proteins were confirmed to be encapsulated into recombinant cytoplasmic polyhedra by Western blotting. Degradation of vaccine proteins was assessed by SDS-PAGE, and the results showed that the encapsulated vaccine proteins in polyhedra could be protected from degradation. Furthermore, when gibel carp were vaccinated with the purified polyhedra from BmNPV-H1-D4ORF-polh and BmNPV-D4ORF-VP3-polh via injection, the antibody titers in the serum of the vaccinated fish reached 1:6400–1:12,800 at 3 weeks post-vaccination. Therelative percentage of survival of immunized gibel carp reached 64.71% and 58.82%, respectively, following challenge with CyHV-2. These results suggest that incorporating vaccine protein into BmCPV polyhedra may be a novel approach for developing aquaculture microencapsulated vaccines.

Massive economic losses have been caused by Hepatopancreas necrosis disease (HPND) to the Chinese mitten crab (Eriocheir sinensis) culture industry; however, the actual cause and pathological mechanisms is unknown. In this study, hepatopancreatic and intestinal cells of the diseased crabs showed distinctive pathological changes which were characteristic of autophagy and apoptosis, but no pathogens were observed. To understand the molecular pathogenesis, a transcriptomic analysis was conducted; approximately 21,523 (26,224) unigenes had COG (KEGG) annotations. The analysis of differentially expressed genes (DEGs) with fold changes > 2 and P < 0.05 between diseased crabs with mild symptoms (XHAF-1) and diseased crabs with severe symptoms (XHAF-2), and crabs without visible symptoms (XHAF-CK) in hepatopancreas showed that DEGs increased with the degree of hepatopancreatic necrosis. The DEGs of XHAF-1 vs XHAF-CK (XHAF-2 vs XHAF-CK) were enriched into 1191 (1366) gene ontology (GO) terms, including the positive regulation of phagocytosis, autophagic vacuole assembly and negative regulation of glial cell apoptotic processes. The DEGs of XHAF-1 vs XHAF-CK (XHAF-2 vs XHAF-CK) were enriched into 32 (45) KEGG pathways including autophagy and the p53 signaling pathway. Furthermore, the expression levels of genes related to autophagy and apoptosis were altered in diseased crabs, suggesting that HPND may be the result of autophagy and apoptosis. Moreover, expression of genes related to retinol metabolism disregulated, implying that the hepatopancreas of crabs with HPND turn from golden yellow/light yellow to almost white were associated with retinol metabolism dysregulation. These findings provide new clues in understanding of the etiology and pathogenesis of HPND.

Hepatopancreatic necrosis syndrome (HPNS) has a severe impact on the Chinese mitten crab (Eriocheir sinensis) industry. In this work, a cross-sectional study method was used to assess the association between 55 variables and HPNS. A total of 98 farmers were surveyed, including 34 whose farms had crabs with HPNS (treated as the diseased group) and 64 whose farms did not have crabs with HPNS (treated as the healthy group). The results demonstrated that 11 risk factors had the greatest effects on HPNS, including “Recent pH in the pond”, “Crabs culture method”, “Current incidence of HPNS in nearby farms”, “Single pond area in the farm”, “Depth of pond's water”, “Frequency of water supplement in the farm”, “Frequency of the abamectin use”, “Frequency of switching aerator on in the farm”, “Frequency of disinfectant use”, “Amount of edible animal ingredient”, “How many factories and residential areas are within 500 m”, “Abundance of Cyanobacteria in the pond”, and “Frequency of clearing the ponds”, suggesting that HPNS prevalence might be closely related to water quality and nutrition. The roles of these 11 variables in the occurrence of HPNS were further evaluated, and it was found that there was a greater risk of HPNS occurrence when the following variables coexisted: “>3 factories and residential areas within 500m” + “high pH” + “>1 disinfectant per shell” + “1.3-2 hm2pond” + “<0.48 g edible animal ingredient per crab” + “No aerator”. Moreover, it was found that temperature might also be an important risk factor for HPNS.

Oral vaccination is a practical method for the active immunization of farmed fish in the matter of animal welfare and handling costs. However, it always shows insufficient protective immunity, mainly due to antigen degradation in the gastrointestinal tract (GIT). Bacillus subtilis spores have been shown to be able to protect surface-display heterologous antigens against degradation. Neverthless, the spores can germinate in GIT, which causes loss of the antigens with spore coat disassembly. Here, we developed a novel surface display system using the B. subtilis spore coat proteins CotB and CotC as anchors for the heterogenous antigen, and the germination-controlling genes cwlJ and sleB as the ectopic integration sites for the fusion genes. Using this display system, we engineered germination-arrest spores displaying the model antigen Vp7 of grass carp reovirus (GCRV) on their surface. Oral vaccination of the engineered spores could confer immune protection against GCRV in grass carp (Ctenopharyngodon idella) via eliciting adaptive humoral and cellular immune responses. Most importantly, the germination-arrest spores were shown to significantly augment immunogenicity and protection above the engineered spores based on the existing surface display system. Therefore, the presently reported antigen expression strategy opens new and promising avenues for developing oral vaccines for the immunization of farmed fish species.

Grass carp reovirus (GCRV) causes grass carp hemorrhage disease and is one of the most damaging factors in the culture of grass carp (Ctenopharyngodon idellus). Until now, the mechanisms underlying GCRV infection have been largely unknown. To explore the molecular responses to GCRV infection in C. idellus kidney (CIK) cells, we used high-throughput sequencing to investigate the circRNA, mRNA, and miRNA expression patterns and the circRNA-miRNA-mRNA network. A total of 76 circRNAs, 798 mRNAs, and 186 miRNAs were identified as differentially expressed (DE) RNAs in GCRV-infected cells compared with uninfected cells, of which 40 circRNAs, 658 mRNAs, and 146 miRNAs were upregulated, and 36 circRNAs, 140 mRNAs, and 40 miRNAs were downregulated (fold change > 2, p < .05). A Gene Ontology (GO) analysis of the DE circRNA parental genes, mRNAs, and miRNA target genes was performed. The parental genes of the DE circRNAs, the DE mRNAs, and the target genes of the DE miRNAs were mainly enriched in metal-ion-binding-, immune-, and DNA-replication-related items, respectively. A KEGG pathway analysis showed that the DE circRNA parental genes, DE mRNAs, and miRNA target genes were predominantly enriched in endocytosis-, immune-, and infection-related pathways, respectively. The miRNA-target interactions were also predicted and competing regulatory networks of endogenous RNAs were constructed based on the DE RNAs. Our prediction results suggest that the circRNA-2780/cik-miRNA-7796/KAT6B, circRNA-1351/cik-miRNA-10648/B2L14 and circRNA-1591/cik-miR-8141/AMPD3 axes may be associated with GCRV infection. This study suggests that the crosstalk between circRNAs and their competing mRNAs play crucial roles in GCRV infection.

To evaluate the genetic variation of offsprings derived from wild and cultivated populations of Macaca thibetana,we analyzed the whole sequence of mitochondrial DNA D-loop region in the 102 individuals of 2 generations of wild (Jinyang region) and cultivated populations (Mabian region).The whole length of the region sequence was 1 091 hp and 115 variable sites were detected.The 102 individuals presented 66 haplotypes,and the 2 generations of Mabian and Jinyang population shared 2 and 1 haplotype with their parents,respectively.The nucleotide polymorphisms of the 2 generations of Mabian population were 0.004 77 and 0.004 09,while those in Jinyang population were 0.002 30 and 0.002 78,indicating no significant difference of genetic diversity occurred between the offsprings and parents of M.thibetana.Genetic distances between the 2 generations of Mabian and Jinyang populations were 0.024 09 and 0.002 55,indicating no significant differentiation happened in the 2 generations in wild or in cultivated situation.NJ tree based on the whole length of D-loop showed 2 independent clades existed in Mabian and Jinyang populations with different generations crossed each other,this demonstrated that neither the random mating nor the artificial selection pressure exerted a significant influence on the genetic structure of the progeny.This study was the first time to use the whole length of mitochondrial DNA D-loop region to analyze the genetic diversity of wild and cultivated Macaca thibetana populations and their offsprings.Our data provide basic support for the standardization of genetic control of experimental animals.

An English idiom is a special combination of words, the meaning of which is not the simple addition of its constituents but a special meaning. English idioms are widely used by English native speakers, which are very important for English as a second language learners. Chinese learners are poor in English idioms comprehension, and they need to adopt a new method for improving their English idioms comprehension. S-S-P-C model means analyzing an English idiom from syntactic, semantic, pragmatic, and cultural perspective, which was applied in a teaching experiment among college students for one semester. The teaching experiment shows that S-S-P-C model is an efficient method for Chinese learners to help improve their English idioms comprehension.

Geographic variation has an important role in both carcinogenesis and prognosis of gastric cancer (GC). High altitude is a special hypoxic environment that is also correlated with the occurrence of GC. Different onset features and prognoses of GC in high altitude with respect to plains are rarely reported and remain unknown. This multicenter study compared different clinicopathological characteristics and prognoses of patients with resected GC who were from locations of both high altitudes and plains in China. From December 2009 to December 2011, patients with resected GC were retrospectively recruited at four centers located at high altitudes and the plains. Clinicopathological data were analyzed to explore the differences between the two groups. The Cox proportional-hazards model was used to investigate the prognostic factors for GC and estimate the independent impact of altitude on long-term survival after adjusting for covariates. Noncardia GC, from a moderate to well tumor grade, was more common in patients from high altitudes. Moreover, a higher proportion of moderate to well and moderate tumor grade and younger age of onset was found in patients with noncardia GC coming from high altitudes. Different overall survival (OS) presented in noncardia GC rather than cardia GC, with 69.94% GC-related 3-yr OS in high altitude versus 75.23% in the plains. High altitude was confirmed as a significant prognostic factor for noncardia GC (the hazard ratio for high altitude vs. plains was 1: 50, with a 95% confidence interval; 1.06-1.82, p = 0.018) through a multivariate Cox proportional-hazards model analysis. This prognostic value was independent of all other factors. High altitude has an important role in clinicopathological features and prognosis of GC. Improvements in GC diagnosis and management at high altitudes are urgently needed.

This paper considers the problem of designing a high-gain sliding-mode-like feedback control law for the model-free tracking of nonholonomic mobile robots. The control approach developed in this work does not need any a-priori knowledge about the model of the robots. A model-free, self-support control algorithm is proposed such that the tracking error can be driven into a pre-specified neighborhood of zero just by using incomplete informations of the moving objects to be tracked. Global stability of the corresponding closed-loop system of tracking error is proved by the Lyapunov stability theory. Finally, the simulation results demonstrate the effectiveness of the proposed controller design method.

This paper considers the problem of designing a visual feedback control law for robust tracking of nonholonomic mobile robots. The control approach developed in this work with uncalibrated visual parameters, unknown control directions, and external disturbances. Using incomplete information of the moving objects to be tracked to propose a model-free, self-support control algorithm to ensure the tracking error can be driven into a prespecified neighborhood of zero. Global stability of the corresponding closed-loop system of tracking error is proved by the Lyapunov stability theory. Finally, the simulation results demonstrate the effectiveness of the proposed controller design method.

This paper considers the stabilization problem of nonholonomic mobile robots with fractional-order sliding-mode controL Two fractional sliding surfaces are designed for the corresponding linear subsystems after using dynamic feedback linearization strategy, then the stabilizing fractional-order controller is proposed, Finally, the simulation results show the efficiency of the control methods.

Although the clinicopathological features of cardia gastric cancer (GC) differ from those of non-cardia GC, it is unclear whether the former has poorer prognosis than the latter. The aim of this study was to compare clinicopathological characteristics and prognosis between cardia and non-cardia GC. From December 2009 to December 2011, 1633 patients who had undergone R0 resection of GC at four Chinese centers were enrolled in this study. Their clinicopathological features and survival outcomes were evaluated. Compared with non-cardia GC, cardia GC was associated with a significantly higher proportion of male patients, older age, more advanced pathological stage, and less-favorable clinicopathological features at diagnosis. The 5-year survival rate was significantly lower in the cardia GC group than in the non-cardia GC group. However, no significant difference in overall survival (OS) was observed between the two groups at any pathological TNM stage. Pathological stage was confirmed as an independent prognostic factor of OS. More advanced disease represents most of the cases in this Chinese population. Compared with patients with non-cardia GC, patients with cardia GC were diagnosed at a more advanced stage and had worse prognosis after undergoing R0 resection. However, cardia and non-cardia GCs have similar stage-for-stage prognoses.

Although studies on the association between the number of lymph nodes resected and prognosis in patients with pT2-4N0 stages of gastric cancer have reported consistent results, there is no consensus on the optimal number of lymph nodes to be examined for pT1N0 stage gastric cancer. The aim of this study was to evaluate the long-term effect of the number of lymph nodes removed on the outcomes of patients with pT1N0 stage gastric cancer after R0 resection.From December 2009 to December 2011, 227 patients undergoing R0 resection of pT1N0 stage gastric cancer at 4 Chinese centers were enrolled in this study. Patients were assigned to 2 groups according to the number of lymph nodes dissected (≤15 or > 15). Standard survival methods and restricted multivariable Cox regression models were applied.More women (P = 0.031) were in the ≤15 group than in the >15 group. The mean number of lymph nodes removed from women was greater than that from men (P = 0.007). The 5-year survival rate was significantly higher in the >15 lymph nodes resected group than the ≤15 group. The number of lymph nodes resected was identified as an independent prognostic factor and was significantly correlated with overall survival (OS).A lymphadenectomy with dissection of more than 15 lymph nodes improved the long-term survival of patients with pT1N0 gastric cancer after R0 resection. Therefore, it is necessary to consider removing more than 15 lymph nodes among such patients.

This paper aims to have an empirical research on college students' language in Guangxi border by the methods of questionnaire and interview. It analyzes the language life of these highly educated young people from the following aspects: language competence, language use; and found that these college students attaches great importance to mandarin, while they begin to downplay the importance of dialect at the same time. Therefore, we should strengthen their importance to the dialect of ideology and vigorously protect the dialect when promoting mandarin policy.