An integrated network based on 2D/3D feature correlations for benign-malignant tumor classification and uncertainty estimation in digital breast tomosynthesis.

Objective.Classification of benign and malignant tumors is important for the early diagnosis of breast cancer. Over the last decade, digital breast tomosynthesis (DBT) has gradually become an effective imaging modality for breast cancer diagnosis due to its ability to generate three-dimensional (3D) visualizations. However, computer-aided diagnosis (CAD) systems based on 3D images require high computational costs and time. Furthermore, there is considerable redundant information in 3D images. Most CAD systems are designed based on 2D images, which may lose the spatial depth information of tumors. In this study, we propose a 2D/3D integrated network for the diagnosis of benign and malignant breast tumors.Approach.We introduce a correlation strategy to describe feature correlations between slices in 3D volumes, corresponding to the tissue relationship and spatial depth features of tumors. The correlation strategy can be used to extract spatial features with little computational cost. In the prediction stage, 3D spatial correlation features and 2D features are both used for classification.Main results.Experimental results demonstrate that our proposed framework achieves higher accuracy and reliability than pure 2D or 3D models. Our framework has a high area under the curve of 0.88 and accuracy of 0.82. The parameter size of the feature extractor in our framework is only 35% of that of the 3D models. In reliability evaluations, our proposed model is more reliable than pure 2D or 3D models because of its effective and nonredundant features.Significance.This study successfully combines 3D spatial correlation features and 2D features for the diagnosis of benign and malignant breast tumors in DBT. In addition to high accuracy and low computational cost, our model is more reliable and can output uncertainty value. From this point of view, the proposed method has the potential to be applied in clinic.

Effects of miR-143 and its target receptor 5-HT2B on agonistic behavior in the Chinese mitten crab (Eriocheir sinensis).

Chinese mitten crab (Eriocheir sinensis) as a commercially important species is widely cultured in China. However, E. sinensis is prone to agonistic behavior, which causes physical damage and wastes energy resources, negatively impacting their growth and survival. Therefore, understanding the regulatory mechanisms that underlie the switching of such behavior is essential for ensuring the efficient and cost-effective aquaculture of E. sinensis. The 5-HT2B receptor is a key downstream target of serotonin (5-HT), which is involved in regulating animal behavior. In this study, the full-length sequence of 5-HT2B gene was cloned. The total length of the 5-HT2B gene was found to be 3127 bp with a 236 bp 5'-UTR (untranslated region), a 779 bp 3'-UTR, and a 2112 bp open reading frame encoding 703 amino acids. Phylogenetic tree analysis revealed that the 5-HT2B amino acid sequence of E. sinensis is highly conserved with that of Cancer borealis. Using in vitro co-culture and luciferase assays, the miR-143 targets the 5-HT2B 3'-UTR and inhibits 5-HT2B expression was confirmed. Furthermore, RT-qPCR and Western blotting analyses revealed that the miR-143 mimic significantly inhibits 5-HT2B mRNA and protein expression. However, injection of miR-143 did not decrease agonistic behavior, indicating that 5-HT2B is not involved in the regulation of such behavior in E. sinensis.

Identification and Integrated Analysis of MicroRNA and mRNA Expression Profiles During Agonistic Behavior in Chinese Mitten Crab (Eriocheir sinensis) Using a Deep Sequencing Approach.

As a commercially important species, the Chinese mitten crab (Eriocheir sinensis) has been cultured for a long time in China. Agonistic behavior often causes limb disability and requires much energy, which is harmful to the growth and survival of crabs. In this paper, we divided crabs into a control group (control, no treatment) and an experimental group (fight, agonistic behavior after 1 h) and then collected the thoracic ganglia (TG) to extract RNA. Subsequently, we first used a deep sequencing approach to examine the transcripts of microRNAs (miRNAs) and messenger RNAs (mRNAs) in E. sinensis displaying agonistic behavior. According to the results, we found 29 significant differentially expressed miRNAs (DEMs) and 116 significant differentially expressed unigenes (DEGs). The DEMs esi-miR-199a-5p, esi-let-7d, esi-miR-200a, and esi-miR-200b might participate in the regulation of agonistic behavior by mediating neuroregulation and energy metabolism. Focusing on the transcripts of the mRNAs, the renin-angiotensin system (RAS) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway might be involved in the regulation of agonistic behavior through glucose metabolism as this pathway was significantly enriched with DEGs. Besides, an integrated analysis of the miRNA and mRNA profiles revealed that the retinoid X receptor (RXR) was also involved in visual signal transduction, which was important for agonistic behavior. In addition, four vital agonistic behavior-related metabolic pathways, including the cAMP signaling, MAPK, protein digestion and absorption, and fatty acid metabolism pathways, were significantly enriched with the predicted target unigenes. In conclusion, the findings of this study might provide important insight enhancing our understanding of the underlying molecular mechanisms of agonistic behavior in E. sinensis.

The protective effects of melatonin on oxidative damage and the immune system of the Chinese mitten crab (Eriocheir sinensis) exposed to deltamethrin.

Deltamethrin (Del), an important broad-spectrum insecticide, is widely used in agricultural activities. However, Del is an effective reactive oxygen species (ROS) inducer that induces oxidative stress damage in cells or tissues. Del is significantly more toxic to aquatic organisms, especially crustaceans, than to mammals and birds. This study was designed to evaluate the protective effect of melatonin (MT) on the toxicity-induced damage of Del after 6 h in Eriocheir sinensis. The results showed that Del exposure significantly induced oxidative damage in the hepatopancreas and mitochondria, with malondialdehyde (MDA) and glutathione (GSH) levels being significantly increased and superoxide dismutase (SOD) activity being significantly decreased. Moreover, Del exposure significantly induced functional damage of the hepatopancreas and mitochondria, with a significant increase in alanine aminotransferase (ALT), aspartate aminotransferase (AST), acid phosphatase (ACP) and alkaline phosphatase (AKP) activities in the hepatopancreas and the ratio of albumin/globulin (ALB/GLB) in serum, which indicated the permeability and integrity of the membranes were damaged and had caused cell damage. In addition, ATP content, Na+-K+-ATPase activity and cytochrome C (Cyt­C) content in mitochondria decreased significantly, which indicated that Del exposure destroyed the normal respiratory chain of mitochondria. We also evaluated the hematological parameters. Although there were no significant differences in total hemocyte count (THC) levels, hemocyte apoptosis was significantly induced by Del exposure, and the hemocyte phagocytic activity and the hemocyanin levels decreased significantly with Del exposure. However, MT pretreatment not only prevented oxidative damage and functional damage caused by Del exposure to the hepatopancreas and mitochondria, but it also restored the hemocyte apoptotic rate and phagocytic activity to normal levels. In short, Del exposure caused significant oxidative and functional damage to the hepatopancreas, mitochondria and hemocytes of E. sinensis, whereas the use of MT almost completely eliminated the damage caused by Del exposure.

The serotonin or dopamine by cyclic adenosine monophosphate-protein kinase A pathway involved in the agonistic behaviour of Chinese mitten crab, Eriocheir sinensis.

Agonistic behaviour is common in an encounter between two crustaceans. It often causes limb disability and consumes a lot of energy, which is harmful for the growth and survival of commercially important crustaceans. In the present study, we mainly focused on the agonistic behaviour of the Chinese mitten crab, Eriocheir sinensis, which is an important species of the aquaculture industry in China. We recorded agnostic behaviour with a high-definition camera and preliminarily evaluated the role of serotonin (5-HT) or dopamine (DA)-cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) pathway and eyestalk in the behaviour. The results showed that agonistic behaviour in E. sinensis consisted of three stages: approach, contact and fight. We found that the number of fights and cumulative time of fight were significantly higher in the male vs. male group than in the female vs. female and female vs. male groups (P < 0.05). After 1 h of agonistic behaviour, 5-HT concentration showed a significant increase and DA concentration showed a significant decrease when compared with the control group (no encounter; P < 0.05). 5-HT1B and 5-HT2B mRNA levels showed a significant increase in the eyestalk (P < 0.05). 5-HT7 mRNA levels showed significant downregulation in the thoracic ganglia and DA1A mRNA levels showed upregulation in the intestine (P < 0.05). DA2 mRNA levels showed a significant decrease in the eyestalk (P < 0.05). These changes were accompanied by a significant increase in cAMP level and significant decrease in PKA level in the haemolymph (P < 0.05). In addition, a significant decrease in glucose levels was detected after the agonistic behaviour. Crustacean hyperglycemic hormone (CHH) mRNA levels showed significant upregulation in the eyestalk and significant downregulation in the intestine (P < 0.05). The number of fights and cumulative time of fight in the left eyestalk ablation (L-X vs. L-X) group were more and longer than those in the intact eyestalk (C vs. C), right eyestalk ablation (R-X vs. R-X) and bilateral eyestalk ablation (D-X vs. D-X) groups. In short, E. sinensis shows special agonistic behaviour modulated by 5-HT or DA-cAMP-PKA pathway and eyestalk, especially the left eyestalk.

5-HT2B, 5-HT7, and DA2 Receptors Mediate the Effects of 5-HT and DA on Agonistic Behavior of the Chinese Mitten Crab (Eriocheir sinensis).

The Chinese mitten crab (Eriocheir sinensis) is a commercially important crab in China and is usually managed at high stocking densities. Agonistic behavior directly impacts crab integrity, survival, and growth and results in economic losses. In the present study, we evaluated the modulatory effects of serotonin (5-HT) and dopamine (DA) though the 5-HT2 and DA2 receptor-cyclic adenosine monophosphate (cAMP)-protein kinase A (PKA) pathway on agonistic behavior. The results showed that injection of either 10-6 mol/crab 5-HT or DA reduced the agonistic behavior of E. sinensis (P < 0.05), as did 10-10 mol/crab DA and 10-8 mol/crab 5-HT and DA (P < 0.05); however, a dose of 10-10 mol/crab 5-HT promoted agonistic behavior. 5-HT significantly increased the mRNA expression level of 5-HT7 receptor and reduced that of the DA2 receptor in the cerebral ganglion (P < 0.05). In contrast to 5-HT, DA significantly decreased 5-HT2B mRNA levels and increased 5-HT7 and DA2 receptor levels in the thoracic ganglia (P < 0.05). In addition, injections of either 5-HT or DA increased the cAMP and PKA levels in hemolymph (P < 0.05). By using in vitro culture of the thoracic ganglia, the current study showed that ketanserin (5-HT2 antagonist) and [R(-)-TNPA] (DA2 agonist) had obvious effects on the expression levels of the two receptors (P < 0.05). In vivo experiments further demonstrated that ketanserin and [R(-)-TNPA] could both significantly reduce the agonistic behavior of the crabs (P < 0.05). Furthermore, both ketanserin and [R(-)-TNPA] promoted the cAMP and PKA levels (P < 0.05). The injection of CPT-cAMP (cAMP analogue) elevated the PKA levels and inhibited agonistic behavior. In summary, this study showed that 5HT-2B and DA2 receptors were involved in the agonistic behavior that 5-HT/DA induced through the cAMP-PKA pathway in E. sinensis.

Changes in bud morphology, growth-related genes and nutritional status during cheliped regeneration in the Chinese mitten crab, Eriocheir sinensis.

During pond culture of Eriocheir sinensis, a high limb-impairment rate restricts the industry development and quality. Therefore, research on limb autotomy and regeneration has important practical significance for the industrial development and basic biology of E. sinensis. This study evaluated the changes in bud morphology, growth-related gene expression and nutritional status during cheliped regeneration in E. sinensis. The study found that the new cheliped was pre-formed in the bud and then regenerated with the completion of molting of E. sinensis. The new cheliped was similar in morphology to the normal cheliped after the first molting but smaller in size. The qRT-PCR results of growth-related genes showed that the expression levels of EcR-mRNA (ecdysteroid receptor) and Chi-mRNA (chitinase) were significantly up-regulated, whereas the expression of MIH-mRNA (molt-inhibiting hormone) was significantly down-regulated (P < 0.05). The nutritional status during the regeneration process showed that the hepatopancreas total lipid content decreased significantly within 28 days and was significantly lower in the autotomy group than in the control group at 14 d and 21 d (P < 0.05). The hepatopancreas fatty acid composition results showed that saturated fatty acids (SFA), highly unsaturated fatty acids (HUFA) and n-3/n-6 were significantly higher in the autotomy group than in the control group at 21 d (P < 0.05), whereas the ∑ n-6 PUFA and ∑ n-3 PUFA at 1 d and 7 d, and the monounsaturated fatty acid (MUFA) at 28 d in the autotomy group were significantly lower than in the control group (P < 0.05). Moreover, the levels of eicosatetraenoic acid (ARA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) showed that DHA was significantly lower at 7 d and significantly higher at 21 d in the autotomy group than in the control group (P < 0.05), whereas ARA and EPA were not significantly different between the two groups. Muscle L-tryptophan content was significantly lower at 1 d and significantly higher at 7 d in the autotomy group than in the control group (P < 0.05). These results indicate that during the cheliped regeneration process, crabs could accelerate molting and regeneration by regulating growth-related gene expression (e.g., EcR-mRNA and MIH-mRNA) and nutrient metabolism (e.g., lipid metabolism).