SULT2B1-CS-DOCK2 axis regulates effector T-cell exhaustion in HCC microenvironment.

BACKGROUND AND AIMS: HCC is a malignant disease. Compared with tyrosine kinase inhibitors (the classical therapy), immune checkpoint inhibitors are more effective in the treatment of HCC, despite their limited efficacy. Among these restricted factors, exhaustion of tumor-infiltrated lymphocytes, especially CD8 + T cells, is a core event. We aimed to determine the key factors contributing to CD8 + T-cell infiltration in HCC and investigate the underlying mechanisms. APPROACH AND RESULTS: Using machine learning and multiplex immunohistochemistry analysis, we showed that dedicator of cytokinesis protein 2 (DOCK2) was a potential indicator of infiltrated CD8 + T cells in HCC. Using RNA sequencing, flow cytometry analysis, and mouse HCC models, we demonstrated that DOCK2 inactivation accounted for infiltrated CD8 + T-cell exhaustion in tumors. Using quasi-targeted metabolomics, mass spectrum, and mass cytometry by time of flight analysis, we found that cholesterol sulfate synthesized by sulfotransferase 2B1 in tumor cells suppressed DOCK2 enzymatic activity of T cells. Through virtual screening, molecular docking simulation, and experiments validation, we demonstrated that tolazamide reversed DOCK2 inactivation-mediated CD8 + T-cell exhaustion and enhanced anti-programmed death-ligand 1 antibody+apatinib immunotherapeutic effects on HCC. CONCLUSIONS: This study indicates that DOCK2 controls CD8 + T-cell infiltration in HCC, and cholesterol sulfate synthesized by sulfotransferase 2B1 in tumor cells promotes effector T-cell exhaustion. The findings suggest that the usage of conventional drugs affects immunotherapy efficacy in HCC patients.

Comparative transcriptome analysis of the gills of Cardisoma armatum provides novel insights into the terrestrial adaptive related mechanism of air exposure stress.

Cardisoma armatum is a typical member of the Gecarcinidae which show significant behavioral, morphological, physiological, and/or biochemical adaptations permitting extended activities on the land. The special gills (branchiostegal lung) of C. armatum play an important role in maintaining osmotic pressure balance and obtaining oxygen to adapt to the terrestrial environment. However, adaptive molecular mechanisms responding to air exposure in C. armatum are still poorly understood. In this study, transcriptomic analysis and histological analysis were conducted on the gills to test adaptive capabilities over 8 h between the aerial exposure (AE) and the water immersion (WI) group. Differentially expressed genes (DEGs) related to terrestrial adaptation were categorized into four broad categories: ion transport, acid-base balance, energy metabolism and immune response. This is the first research to reveal the molecular mechanism of terrestrial adaptation in C. armatum, and will provide new insight into the molecular genetic basis of terrestrial adaptation in crabs.

Comparative transcriptome analysis of the gills of Procambarus clarkii provide novel insights into the response mechanism of ammonia stress tolerance.

Procambarus clarkii is an important model crustacean organism in many researches. Ammonia nitrogen is one of common contaminants in aquatic environment, influencing the health of aquatic organisms. The primary objective of this study was to investigate molecular mechanisms on ammonia stress in gills of P. clarkii to provide new insights into the strategies of aquatic animals in responding to high concentration of ammonia in the environment. Procambarus clarkii were randomly assigned into two groups (ammonia stress group, AG; control group, CG), and gill samples were dependently excised from AG and CG. Then response mechanisms on ammonia stress were investigated based on transcriptome data of P. clarkii. 9237 differentially expressed genes were identified in ammonia stress group. The genes of ion transport enzymes (NKA and SLC6A5S) were significantly up-regulated. Whereas the immune-related genes (e.g. MAP3K7, HSP70, HSP90A, CTSF, CTSL1, CHI and CTL4) and pathways were significantly up-regulated, which played an important role in reacting to ammonia stress. Procambarus clarkii may enhance immune defense to counteract ammonia toxicity by the up-regulation of immune-related genes and signaling pathways. The activities of ion transport enzymes are changed to mobilise signal transduction and ion channel regulation for adapting to ammonia environment. These previous key genes play an important role in resistance to ammonia stress to better prepare for survival in high concentration of ammonia.

Toxic effects of metal copper stress on immunity, metabolism and pathologic changes in Chinese mitten crab (Eriocheir japonica sinensis).

Copper (Cu2+), which represents a major physiological challenge for crab culture, is ubiquitous in the aquatic culture environment, and gills are the first organs that come into direct contact with the environment. However, the molecular basis of the response of crabs to Cu2+ stress remains unclear. Here, we conducted a transcriptome and differential expression analysis on the gills from Chinese mitten crab unexposed and exposed to Cu2+ for 24 h. The comparative transcriptome analysis identified 2486 differentially expressed genes (DEGs). GO functional analysis and KEGG pathway analysis revealed some DEGs, which were mostly related to immunity, metabolism, osmotic regulation, Cu2+ homeostasis regulation, antioxidant activity, and detoxification process. Some pathways related to humoral and cellular immunity, such as phagosome, peroxisome, lysosome, mTOR signaling pathway, PI3K-Akt signaling pathway, Toll-like receptor signaling pathway, and T cell receptor signaling pathway were enhanced under Cu2+ stress. In addition, Cu2+ stress altered the expression patterns of key phagocytosis and apoptosis genes (lectin, cathepsin L, Rab7, and HSP70), confirming that Cu2+ can induce oxidative stress and eventually even apoptosis. Histological analysis revealed that the copper can induce damage at the cellular level. This comparative transcriptome analysis provides valuable molecular information to aid future study of the immune mechanism of Chinese mitten crab in response to Cu2+ stress and provides a foundation for further understanding of the effects of metal toxicity.

Evolution of digestive enzyme genes associated with dietary diversity of crabs.

Crabs feed on a wide range of items and display diverse feeding strategies. The primary objective of this study was to investigate 10 digestive enzyme genes in representative crabs to provide insights into the genetic basis of feeding habits among crab functional groups. Crabs were classified into three groups based on their feeding habits: herbivores (HV), omnivores (OV), and carnivores (CV). To test whether crabs' feeding adaptations matched adaptive evolution of digestive enzyme genes, we examined the 10 digestive enzyme genes of 12 crab species based on hepatopancreas transcriptome data. Each of the digestive enzyme genes was compared to orthologous sequences using both nucleotide- (i.e., PAML and Datamonkey) and protein-level (i.e., TreeSAAP) approaches. Positive selection genes were detected in HV crabs (AMYA, APN, and MGAM) and CV crabs (APN, CPB, PNLIP, RISC, TRY, and XPD). Additionally, a series of positive selection sites were localized in important functional regions of these digestive enzyme genes. This is the first study to characterize the molecular basis of crabs' digestive enzyme genes based on functional feeding group. Our data suggest that HV crabs have evolved an enhanced digestion capacity for carbohydrates, and CV crabs have acquired digestion capacity for proteins and lipids.

Comparative transcriptome analysis of the gills of Procambarus clarkii provides novel insights into the immune-related mechanism of copper stress tolerance.

The red-swamp crayfish (Procambarus clarkii) is the most important economic shrimp species in China, and is an important model crustacean organism in many fields of research. In crustaceans, gills interface directly with the ambient environment and thus play a vital role in the toxicology. In the context of increasing environmental heavy metal pollution, the relationship between copper (Cu2+) stress and the immune response of P. clarkii has recently received considerable attention. However, impact of Cu2+ on the crayfish immune system is still not fully understood. In this study, we used Illumina sequencing technology to perform a transcriptome analysis of the gills of P. clarkii after 24 h of Cu2+ treatment. A total of 37,226,812 unigenes were assembled, and 1943 unigenes were significantly differentially expressed between the control and Cu2+ treatment groups. Functional categorization of differentially expressed genes (DEGs) revealed that genes related to antioxidant activity, detoxication, metabolic processes, biosynthetic processes, and immune system processes were differentially regulated during Cu2+ stress. In addition, DEGs in the immune system were classified as being related to the MAPK signaling pathway, purine metabolism, Toll and Imd signaling pathway, PI3K-Akt signaling pathway and Hippo signaling pathway. Five genes (CuZnSOD, CAT, IDH1, PHYH and DECR2) were significantly up-regulated in the peroxisome pathway, which plays an important role in reacting to oxidative stress. Importantly, qRT-PCR validation of the results for seven genes chosen at random (NDK, ATP6L, ATP5C1, RPS14, RPL22e, CTSF and HSP90A) confirmed the Illumina sequencing results. This study provides a valuable starting point for further studies to elucidate the molecular basis of the immune system's response to Cu2+ stress in crayfish.

Di-(2-ethylhexyl) Phthalate Exposure Modulates Antioxidant Enzyme Activity and Gene Expression in Juvenile and Adult Daphnia magna.

Di-(2-ethylhexyl) phthalate (DEHP) is a plasticizer used in the polyvinyl chloride industry worldwide. DEHP exists in the aquatic environments for decades. However, the toxicological effects of DEHP to aquatic organisms have not been adequately researched. We investigated acute toxicity, oxidative damage, antioxidant enzyme activities, and gene expression patterns of antioxidant enzymes in juvenile and adult Daphnia magna exposed to DEHP. We found that the median lethal concentrations (LC50) of DEHP for juveniles exposed for 24 and 48 h were 0.83 and 0.56 mg L-1, respectively. The LC50 of DEHP in adults exposed for 24 and 48 h were 0.48 and 0.35 mg L-1. Daphnia magna that was exposed to DEHP had increased malondialdehyde levels for 24 h and lower total antioxidant capacity compared with the control. Activity levels of antioxidant enzymes superoxide dismutase and phase II detoxifying enzyme glutathione S-transferases were significantly higher upon initial exposure for 24 h, and enzyme activity was then diminished at high concentrations and prolonged exposure for 48 h. Gene expression levels of cat and gst were notably reduced or increased upon DEHP exposure. These findings suggest that DEHP can cause biochemical and physiological effects in juvenile and adult D. magna by inhibiting enzymes, an increase in lipid peroxidation levels and changes both transcription levels of enzymes (cat, gst). On the whole, juveniles and adults both responded similarly to DEHP. Our findings will contribute to the understanding of toxic mechanisms in phthalate esters and the evaluation of environmental risks in aquatic ecosystems.

Characterization of Acute and Chronic Toxicity of DBP to Daphnia magna.

As a commonly used phthalate compound, di(n-butyl) phthalate (DBP) is an emerging group of polyvinyl chloride plasticizers. The acute toxicity of DBP has been extensively studied using the aquatic indicator organism, Daphnia magna. However, little is known about chronic and transgenerational toxicity of DBP. In this study, acute LC50 values were 3.04 mg/L (24 h) and 2.55 mg/L (48 h). Chronic toxicity tests in the case of maternal exposure to DBP revealed that DBP had negligible effects on growth and reproduction of F3 generation of D. magna, although the growth rate of body length and the intrinsic rate of increase were prominently reduced, to a pretty small extent. At specific concentrations, DBP generated beneficial effects on the parental generation of D. magna and no obvious impacts on the F1 generation. This study showed that maternal exposure to DBP did not cause any transgenerational effects on D. magna.

A Middle Triassic thoracopterid from China highlights the evolutionary origin of overwater gliding in early ray-finned fishes.

Gliding adaptations in thoracopterid flying fishes represent a remarkable case of convergent evolution of overwater gliding strategy with modern exocoetid flying fishes, but the evolutionary origin of this strategy was poorly known in the thoracopterids because of lack of transitional forms. Until recently, all thoracopterids, from the Late Triassic of Austria and Italy and the Middle Triassic of South China, were highly specialized 'four-winged' gliders in having wing-like paired fins and an asymmetrical caudal fin with the lower caudal lobe notably larger than the upper lobe. Here, we show that the new genus Wushaichthys and the previously alleged 'peltopleurid' Peripeltopleurus, from the Middle Triassic (Ladinian, 235-242 Ma) of South China and near the Ladinian/Anisian boundary of southern Switzerland and northern Italy, respectively, represent the most primitive and oldest known thoracopterids. Wushaichthys, the most basal thoracopterid, shows certain derived features of this group in the skull. Peripeltopleurus shows a condition intermediate between Wushaichthys and Thoracopterus in having a slightly asymmetrical caudal fin but still lacking wing-like paired fins. Phylogenetic studies suggest that the evolution of overwater gliding of thoracopterids was gradual in nature; a four-stage adaption following the 'cranial specialization-asymmetrical caudal fin-enlarged paired fins-scale reduction' sequence has been recognized in thoracopterid evolution. Moreover, Wushaichthys and Peripeltopleurus bear hooklets on the anal fin of supposed males, resembling those of modern viviparious teleosts. Early thoracopterids probably had evolved a live-bearing reproductive strategy.

Cold threat and moisture deficit induced individual tree mortality via 25-year monitoring in seminatural mixed forests, northeastern China.

Accurately predicting tree mortality in mixed forests sets a challenge for conventional models because of large uncertainty, especially under changing climate. Machine learning algorithms had potential for predicting individual tree mortality with higher accuracy via filtering the relevant climatic and environmental factors. In this study, the sensitivity of individual tree mortality to regional climate was validated by modeling in seminatural mixed coniferous forests based on 25-year observations in northeast of China. Three advanced machine learning and deep learning algorithms were employed, including support vector machines, multi-layer perceptron, and random forests. Mortality was predicted by the effects of multiple inherent and environmental factors, including tree size and growth, topography, competition, stand structure and regional climate. All three types of models performed satisfactorily with their values of the areas under receiving operating characteristic curve (AUC) > 0.9. With tree growth, competition and regional climate as input variables, a model based on random forests showed the highest values of the explained variance score (0.862) and AUC (0.914). Since the trees were vulnerable despite their species, mortality could occur after growth limit induced by insufficient or excessive sun radiation during growing seasons, cold threat caused thermal insufficiency in winters, and annual moisture constraints in these mixed coniferous forests. Our findings could enrich basic knowledge on individual tree mortality caused by water and heat inadequacy with the negative impacts of global warming. Successful individual tree mortality modeling via advanced algorithms in mixed forests could assist in adaptive forest ecology modeling in large areas.

The multi-omics analysis in the hepatopancreas of Eriocheir sinensis provides novel insights into the response mechanism of heat stress.

Under global warming, heat stress can induce the excessive production of reactive oxygen species, causing irreversible damage to aquatic animals. It is essential to predict potentially harmful impacts on aquatic organisms under heat stress. Eriocheir sinensis, a typical crustacean crab, is widely distributed in China, American and Europe. Parent E. sinensis need migrate to the estuaries to reproduce in winter, and temperature is a key environmental factor. Herein, we performed a comprehensive transcriptomic and proteomic analysis in the hepatopancreas of E. sinensis under heat stress (20 °C and 30 °C), focusing on heat shock protein family, antioxidant system, energy metabolism and immune defense. The results revealed that parent E. sinensis generated adaptative responses to maintain physiological function under 20 °C stress via the transcriptional up-regulation of energy metabolism enzymes, mRNA synthesis and heat shock proteins. The transcriptional inhibition of key enzymes related to energy metabolism implied that 30 °C stress may lead to the dysfunction of energy metabolism in parent E. sinensis. Meanwhile, parent E. sinensis also enhanced the expression of ferritin and phospholipase D at translational level, and the glutathione s-transferase and heat shock protein 70 at both transcriptional and translational levels, speculating that parent E. sinensis can strengthen antioxidant and immune capacity to resist oxidative stress under 30 °C stress. This study elucidated the potential molecular mechanism in response to heat stress of parent E. sinensis hepatopancreas. The preliminary selection of heat tolerance genes or proteins in E. sinensis can provide a reference for the population prediction and the study of evolutionary mechanism under heat stress in crabs.

The Influence of Shelter Type and Coverage on Crayfish (Procambarus clarkii) Predation by Catfish (Silurus asotus): A Controlled Environment Study.

Procambarus clarkii is adept at using natural shelters and caves to evade attacks from predators. However, the concealment abilities and mechanisms of P. clarkii for different types of shelters under predation pressure have not yet been reported. In this study, laboratory experiments were carried out to determine the effects of different coverages (25%, 50%, and 75%) and different combinations (I-VII) of three types of shelters (PVC pipes, water grass, and stone) on the predation rhythm, behavior, and abilities of Silurus asotus on P. clarkii. The results indicated that the predation of S. asotus on P. clarkii exhibited significant rhythmicity under shelter conditions, excluding PVC pipes, 75% stone, and combination VI. Among the three types of shelters, PVC pipes provided the strongest concealment, followed by stone and water grass. With the increase in shelter coverage, the anti-predation ability of P. clarkii continued to increase, and the optimal shade rate for water grass was 50%. In the different shelter combinations, the environmental complexity had little effect on the predation activity of S. asotus on P. clarkii. These findings demonstrated that the type and abundance of shelters in the wild environment can affect the predation rhythm and activities of S. asotus on P. clarkii.

Correlation analysis of migration in Eriocheir sinensis of the Yangtze River estuary based on salinity preference.

Reproduction and development are the most important stages in the life of aquatic animals, which are also the most sensitive stages to environmental stress. As a migratory crab, parent Eriocheir sinensis need to migrate to estuary (brackish water) for reproduction, and megalopas need to migrate to freshwater for development. Yangtze River estuary is located at the junction of the Yangtze River and the East China Sea, and the salinity of different regions varies greatly (salinity 0-30‰). Therefore, salinity is one of the key environmental factors affecting E. sinensis. In this study, the salinity preference of parent and megalopas E. sinensis was observed and analyzed by the six-chambered and seven-chambered devices of laboratory experiments. Two groups of alternative salinity were set for parent crabs (7, 14, 21, 28, and 35 ‰) and megalopas (0, 5, 10, 15, 20, 25, and 30‰). The results indicated that males showed no obvious preference for salinity, while females had obvious preference for different salinity and stronger dependence and tolerance to saltwater. Megalopas showed a significant preference for low salinity which was beneficial to migrating to freshwater quickly. It was speculated that the behavior of megalopas tending to freshwater was an inherited behavior in the evolutionary process rather than influenced by environment. The salinity preference of E. sinensis might change the position of spawning grounds and migratory routes, and then the annual fluctuation of salinity in Yangtze River estuary will affect the change of E. sinensis resources.

Integration of Transcriptomics and Proteomics Analysis Reveals the Molecular Mechanism of Eriocheir sinensis Gills Exposed to Heat Stress.

Heat stress is an increasingly concerning topic under global warming. Heat stress can induce organisms to produce excess reactive oxygen species, which will lead to cell damage and destroy the antioxidant defense of aquatic animals. Chinese mitten crab, Eriocheir sinensis, is sensitive to the change in water temperature, and parent crabs are more vulnerable during the breeding stage. In the present study, the multi-omics responses of parent E. sinensis gills to heat stress (24 h) were determined via transcriptome and proteome. The integrative analysis revealed that heat shock protein 70 (HSP70) and glutathione s-transferase (GST) were significantly up-regulated at gene and protein levels after heat stress, indicating that HSP70 and the antioxidant system participated in the regulatory mechanism of heat stress to resist oxidative damage. Moreover, the "Relaxin signaling pathway" was also activated at gene and protein levels under 30 °C stress, which implied that relaxin may be essential and responsible for reducing the oxidative damage of gills caused by extreme heat stress. These findings provided an understanding of the regulation mechanism in E. sinensis under heat stress at gene and protein levels. The mining of key functional genes, proteins, and pathways can also provide a basis for the cultivation of new varieties resistant to oxidative stress.

Phosphorescence enhancement of pyridinium macrocycles by poly(vinylalcohol).

A phosphorescence enhancement of pyridinium macrocycle/monomer phosphors is realized with up to 14.7-fold prolonging of the phosphorescence lifetimes and visible afterglow by doping into a poly(vinylalcohol) (PVA) matrix. The abundant hydrogen-bonding interactions and electrostatic interactions between the phosphors and the PVA suppressed the nonradiative decay processes, slowed down the radiative decay and nonradiative decay of triplet states, and therefore promoted the long-lived RTP.

BMI1-induced CD127+KLRG1+ memory T cells enhance the efficacy of liver cancer immunotherapy.

The efficacy of HCC (hepatocellular carcinoma) immunotherapy is hindered by the limited reactivity and short duration of tumor-infiltrating T cells. These deficiencies may be ascribed to the proliferative ability of T cells. The primary objective of this study was to identify the key factor regulating tumor-infiltrating lymphocytes (TIL) proliferation within the HCC microenvironment. Through the utilization of tissue-infiltrated T cell proteomics and fraction proteomics, we analyzed the differential proteins in T cells among HCC, liver fibrosis, and hemangioma (serving as controls) groups. Additionally, we examined the differential regulatory TFs of T cells between the HCC and VH (volunteer healthy, as a control) groups. Using cyTOF and flow cytometry technologies, as well as generating CD8+ T-specific BMI1 knockout mice, we confirmed that BMI1 controls CD127+KLRG1+ memory cell differentiation. Through RNA-seq and MeRIP-seq, we verified that BMI1 regulates TCF1 expression independently of its classical function. Furthermore, by conducting Tyramide signal amplification (TSA) IHC analysis, employing a hydrodynamic mouse HCC model, and utilizing liver-specific nanoparticle targeting therapy, we demonstrated that BMI1 in HCC influences the proliferation of infiltrating CD8+T. BMI1 inhibition promotes effector T cell differentiation while suppressing memory T cell differentiation. Moreover, liver-specific BMI1 knockdown proves beneficial in ameliorating T cell dysfunction and decelerating HCC progression. Our research group has pioneered the exploration of the proteomics of HCC-infiltrated T cells, shedding light on the pivotal role of BMI1 in controlling CD127+KLRG1+ memory CD8+ T cell differentiation, which serves as the cornerstone for achieving immunotherapy efficacy in HCC.

The mechanisms to dispose of misfolded proteins in the endoplasmic reticulum of adipocytes.

Endoplasmic reticulum (ER)-associated degradation (ERAD) and ER-phagy are two principal degradative mechanisms for ER proteins and aggregates, respectively; however, the crosstalk between these two pathways under physiological settings remains unexplored. Using adipocytes as a model system, here we report that SEL1L-HRD1 protein complex of ERAD degrades misfolded ER proteins and limits ER-phagy and that, only when SEL1L-HRD1 ERAD is impaired, the ER becomes fragmented and cleared by ER-phagy. When both are compromised, ER fragments containing misfolded proteins spatially coalesce into a distinct architecture termed Coalescence of ER Fragments (CERFs), consisted of lipoprotein lipase (LPL, a key lipolytic enzyme and an endogenous SEL1L-HRD1 substrate) and certain ER chaperones. CERFs enlarge and become increasingly insoluble with age. Finally, we reconstitute the CERFs through LPL and BiP phase separation in vitro, a process influenced by both redox environment and C-terminal tryptophan loop of LPL. Hence, our findings demonstrate a sequence of events centered around SEL1L-HRD1 ERAD to dispose of misfolded proteins in the ER of adipocytes, highlighting the profound cellular adaptability to misfolded proteins in the ER in vivo.

Preference for Shelters at Different Developmental Stages of Chinese Mitten Crab (Eriocheir sinensis).

A shelter is a good habitat for aquatic organisms, which could aid in avoiding cannibalism and facilitate predation. Chinese Mitten Crab (Eriocheir sinensis) is an important aquaculture species with troglodytism and nocturnal habit. To clarify the preference for shelters at different developmental stages of E. sinensis, different shelters (mud, sand, grass and rocks) were selected for comparison. These results indicated that juvenile crabs had a significant preference for grass; button-sized crabs preferred to hide in mud; and the favorite shelters for parent crabs were rocks, followed by mud. E. sinensis in three stages all showed concealing behavior. The concealing behavior of juvenile crabs was the most significant, followed by button-sized and parent crabs. Additionally, E. sinensis held a low hiding rate at night but a high hiding rate during the daytime due to nocturnal habits. These findings will help to better understand the habits of E. sinensis and provide a reference for resource restoration, habitat construction and the restoration of E. sinensis.

Transcriptome analysis of the gills of Eriocheir sinensis provide novel insights into the molecular mechanisms of the pH stress response.

Eriocheir sinensis is an important economic species in China, which is easily affected by pH changes. However, the molecular mechanism of the pH stress response in E. sinensis is still unclear. Therefore, this study aimed to examine the molecular response mechanism of E. sinensis based on pH variation surveillance, histopathological evaluation and transcriptomic analyses. Firstly, pH variation surveillance showed that E. sinensis could actively regulate the pH of its environment. Meanwhile, the histopathological evaluation suggested that pH stress seriously damaged the gills, especially at high pH. Finally, transcriptome analysis showed that the expression of genes related to ion transport, immune stress, and energy metabolism significantly changed. Many genes played an important role in the pH response of E. sinensis, such as carbonic anhydrase (CA), mitochondrial proton/calcium exchanger protein (LETM1), recombinant sodium/hydrogen exchanger 3 (SLC9A3/NHE3), heat shock protein 90 alpha family class a member (HSP90A), alkylglycerone phosphate synthase (AGPS), succinate-CoA ligase ADP-forming subunit beta (LSC2), and superoxide dismutase (SOD). Our study revealed the molecular response mechanism of E. sinensis in response to pH stress, thus providing a basis for further research on the molecular mechanism of response to pH stress in aquatic animals.

Salinity-Driven Changes in Behavioral Responses of Catadromous Eriocher sinensis.

The effects of salinity on behavior are far-reaching, and Eriocheir sinensis showed disparate behaviors under different salinity conditions. Female crabs were more active in saline water, especially low salinity stress, which is beneficial for female crabs to escape from the low-salinity environment quickly. Then, antennal movement indicated that antennae might be the main osmoreceptors in E. sinensis, and 65 min might be a good starting time for salinity stress to analyze osmoregulation in crabs. Interestingly, E. sinensis had obvious behavioral differences in the high and low salinity stress, and behaviors were more intense in a salinity dip from salinity 18 to salinity 0. This study analyzed the osmoregulatory process of catadromous E. sinensis in different salinity from the point of osmoregulatory organ and behavioral response. These results will provide a scientific basis for the osmoregulatory mechanism of E. sinensis, which are conducive to evaluating and analyzing the impact of saltwater intrusion in the Yangtze River estuary on resource fluctuation.