Discovery of Nine Dipeptidyl Peptidase-4 Inhibitors from Coptis chinensis Using Virtual Screening, Bioactivity Evaluation, and Binding Studies.

The objective of this study was to identify multiple alkaloids in Coptis chinensis that demonstrate inhibitory activity against DPP-4 and systematically evaluate their activity and binding characteristics. A combined strategy that included molecular docking, a DPP-4 inhibition assay, surface plasmon resonance (SPR), and a molecular dynamics simulation technique was employed. The results showed that nine alkaloids in Coptis chinensis directly inhibited DPP-4, with IC50 values of 3.44-53.73 µM. SPR-based binding studies revealed that these alkaloids display rapid binding and dissociation characteristics when interacting with DPP-4, with KD values ranging from 8.11 to 29.97 µM. A molecular dynamics analysis revealed that equilibrium was rapidly reached by nine DPP-4-ligand systems with minimal fluctuations, while binding free energy calculations showed that the ∆Gbind values for the nine test compounds ranged from -31.84 to -16.06 kcal/mol. The most important forces for the binding of these alkaloids with DPP-4 are electrostatic interactions and van der Waals forces. Various important amino acid residues, such as Arg125, His126, Phe357, Arg358, and Tyr547, were involved in the inhibition of DPP-4 by the compounds, revealing a mechanistic basis for the further optimization of these alkaloids as DPP-4 inhibitors. This study confirmed nine alkaloids as direct inhibitors of DPP-4 and characterized their binding features, thereby providing a basis for further research and development on novel DPP-4 inhibitors.

Transcriptomic insights into cessation of clam embryonic development following transgenerational exposure to ocean acidity extreme.

Ocean acidity extremes (OAX) events are becoming more frequent and intense in coastal areas in the context of climate change, generating widespread consequences on marine calcifying organisms and ecosystems they support. While transgenerational exposure to end-of-century scenario of ocean acidification (i.e., at pH 7.7) can confer calcifiers resilience, whether and to what extent such resilience holds true under OAX conditions is still poorly understood. Here, we found that transgenerational exposure of Ruditapes philippinarum to OAX resulted in cessation of embryonic development at the trochophore stage, implying devastating consequences of OAX on marine bivalves. We identified a large number of differentially expressed genes in embryos following transgenerationally exposed to OAX, which were mainly significantly enriched in KEGG pathways related to energy metabolism, immunity and apoptosis. These pathways were significantly activated, and genes involved in these processes were up-regulated, indicating strong cellular stress responses to OAX. These findings demonstrate that transgenerational exposure to OAX can result in embryonic developmental cessation by severe cellular damages, implying that transgenerational acclimation maybe not a panacea for marine bivalves to cope with OAX, and hence urgent efforts are required to understand consequences of intensifying OAX events in coastal ecosystems.

Resistance of an intertidal oyster(Saccostrea mordax)to marine heatwaves and the implication for reef building.

Marine heatwaves (MHWs) have a significant impact on intertidal bivalves and the ecosystems they sustain, causing the destruction of organisms' original habitats. Saccostrea mordax mainly inhabits the intertidal zone around the equator, exhibiting potential tolerance to high temperatures and maybe a species suitable for habitat restoration. However, an understanding about the tolerance mechanism of S. mordax to high temperatures is unclear. It is also unknown the extent to which S. mordax can tolerate repeated heatwaves of increasing intensity and frequency. Here, we simulated the effects of two scenarios of MHWs and measured the physiological and biochemical responses and gene expression spectrum of S. mordax. The predicted responses varied greatly across heatwaves, and no heatwave had a significant impact on the survival of S. mordax. Specifically, there were no statistically significant changes apparent in the standard metabolic rate and the activities of enzymes of the oyster during repeated heatwaves. S. mordax exposed to high-intensity heatwaves enhanced their standard metabolic rate to fuel essential physiological maintenance and increasing activity of SOD and expression of HSP70/90. These strategies are presumably at the expense of functions related to immunity and growth, as best exemplified by significant depressions in activities of enzymes (NaK, CaMg, T-ATP, and AKP) and expression levels of genes (Rab, eEF-2, HMGR, Rac1, SGK, Rab8, etc.). The performance status of S. mordax tends to improve by implementing a suite of less energy-costly compensatory mechanisms at various levels of biological organization when re-exposed to heatwaves. The adaptive abilities shown by S. mordax indicate that they can play a crucial role in the restoration of oyster reefs in tropical seas.

Behavioral responses of intertidal clams to compound extreme weather and climate events.

Rapidly increasing concentration of carbon dioxide (CO2) in the atmosphere not only results in global warming, but also drives increasing seawater acidification. Infaunal bivalves play critical roles in benthic-pelagic coupling, but little is known about their behavioral responses to compound climate events. Here, we tested how heatwaves and acidification affected the behavior of Manila clams (Ruditapes philippinarum). Under acidified conditions, the clams remained capable of burrowing into sediments. Yet, when heatwaves attacked, significant decreases in burrowing ability occurred. Following two consecutive events of heatwaves, the clams exhibited rapid behavioral acclimation. The present study showed that the behavior of R. philippinarum is more sensitive to heatwaves than acidification. Given that the behavior can act as an early and sensitive indicator of the fitness of intertidal bivalves, whether, and to what extent, behavioral acclimation can persist under scenarios of intensifying heatwaves in the context of ocean acidification deserve further investigations.

The Role of the TLR4-MyD88 Signaling Pathway in the Immune Response of the Selected Scallop Strain "Hongmo No. 1" to Heat Stress.

The innate immunity of marine bivalves is challenged upon exposure to heat stress, especially with increases in the frequency and intensity of heat waves. TLR4 serves a classical pattern recognition receptor in recognizing pathogenic microorganisms and activating immune responses. In this study, three genes, HMTLR4, HMMyD88 and HMTRAF6, were characterized as homologs of genes in the TLR4-MyD88 signaling pathway in the selected scallop strain "Hongmo No. 1". According to RT-PCR, acute heat stress (32 °C) inhibited genes in the TLR4-MyD88 signaling pathway, and LPS stimulation-induced activation of TLR4-MyD88 signal transduction was also negatively affected at 32 °C. ELISA showed LPS-induced tumor necrosis factor alpha (TNF-α) or lysozyme (LZM) activity, but this was independent of temperature. RNA interference (RNAi) confirmed that HMTLR4 silencing suppressed the expression of its downstream gene, whether at 24 °C or at 32 °C. The level of TNF-α and the activity of LZM also decreased after injection with dsRNA, indicating a negative effect on the innate immunity of scallops. Additionally, acute heat stress affected the suppression of downstream gene expression when compared with that at 24 °C, which led us to the hypothesis that heat stress directly influences the downstream targets of HMTLR4. These results enrich the knowledge of scallop immunity under heat stress and can be beneficial for the genetic improvement of new scallop strains with higher thermotolerance.

Mapping conservation priorities for wild yak (Bos mutus) habitats on the Tibetan Plateau, China.

The wild yak (Bos mutus) is a cold-tolerant herbivore native to the Tibetan Plateau and has been categorized as vulnerable by the International Union for Conservation of Nature and Natural Resources. Low population densities within currently fragmented habitats and unclear landscape conservation priorities warrant attention. Herein, we employed the maximum entropy (MaxEnt) model using over 900 wild yak occurrence records to model wild yak habitat suitability. Our analysis revealed unprotected wild yak landscapes covering 30.79 % of the habitat area, indicating a conservation gap between protected areas (PAs) and wild yak habitats. To protect metapopulation dynamics and mitigate high risks of poaching, habitat degradation and fragmentation, resource competition, and degenerated genetic characterization of wild yaks in fragmented and degraded habitat, we identified eight habitat patches as landscape conservation units (LCUs) and 14 linkages among the LCUs, enhancing the connectivity between LCUs to decrease negative effects of genetic threats. A centrality analysis demonstrated that Changtang, Arjinshan, and Hoh Xil national nature reserves and their linkages are all critical for the maintenance of habitat connectivity. Here, we suggest that habitat- and LCU-specific conservation strategies should be highlighted during the establishment of PAs and transboundary cooperation. Ultimately, our results can assist conservationists and land managers in comprehending wild yak distribution, movement, and habitat requirements, as well as for the development of effective protection strategies. Furthermore, the combined modeling method (MaxEnt-Zonation-InVEST) could be utilized as a component for identifying conservation priorities and linkages between core patches for species and assessing the efficiency of PAs, core habitats, and corridors in achieving conservation goals. Our study can provide a framework in identifying priority conservation and connectivity between habitat patches to facilitate effectively conservation and genetic resilience for endangered species in fragmented habitats.

Specific molecular weight of Lycium barbarum polysaccharide for robust breast cancer regression by repolarizing tumor-associated macrophages.

The pro-tumorigenic M2-type tumor-associated macrophages (TAMs) in the immunosuppressive tumor microenvironment (TME) promote the progression, angiogenesis, and metastasis of breast cancer. The repolarization of TAMs from an M2-type toward an M1-type holds great potential for the inhibition of breast cancer. Here, we report that Lycium barbarum polysaccharides (LBPs) can significantly reconstruct the TME by modulating the function of TAMs. Specifically, we separated four distinct molecular weight segments of LBPs and compared their repolarization effects on TAMs in TME. The results showed that LBP segments within 50-100 kDa molecular weight range exhibited the prime effect on the macrophage repolarization, augmented phagocytosis effect of the repolarized macrophages on breast cancer cells, and regression of breast tumor in a tumor-bearing mouse model. In addition, RNA-sequencing confirms that this segment of LBP displays an enhanced anti-breast cancer effect through innate immune responses. This study highlights the therapeutic potential of LBP segments within the 50-100 kDa molecular weight range for macrophage repolarization, paving ways to offer new strategies for the treatment of breast cancer.

Exploring the regulatory role of Linc00893 in asthenozoospermia: Insights into sperm motility and SSC viability.

The role of long intergenic noncoding RNA 00893 (Linc00893) in asthenozoospermia (AS) and its impact on sperm motility remains unclear The present study explored the effect of Linc00893 on AS, specifically its effect on sperm motility and its relationship with spermatogonial stem cell (SSC) vitality and myosin heavy chain 9 (MYH9) protein expression. Linc00893 expression was analyzed in semen samples using reverse transcription­quantitative PCR, revealing a significant downregulation in samples from individuals with AS compared with those from healthy subjects. This downregulation was found to be negatively correlated with parameters of sperm motility. To further understand the role of Linc00893, small interfering RNA was used to knockdown its expression in SSCs. This knockdown led to a marked decrease in cell vitality and an increase in apoptosis. Notably, Linc00893 knockdown was shown to inhibit MYH9 expression by competitively binding with microRNA­107, a finding verified by dual­luciferase reporter and RNA immunoprecipitation assays. Furthermore, using the GSE160749 dataset from the Gene Expression Omnibus database, it was revealed that MYH9 protein expression was downregulated in AS samples. Subsequently, lentiviral vectors were constructed to induce overexpression of MYH9, which in turn reduced SSC apoptosis and counteracted the apoptosis triggered by Linc00893 knockdown. In conclusion, the present study identified the role of Linc00893 in AS, particularly its regulatory impact on sperm motility, SSC vitality and MYH9 expression. These findings may provide information on the potential regulatory mechanisms in AS development, and identify Linc00893 and MYH9 as possible targets for diagnosing and treating AS­related disorders.

Sex-specific responses of Ruditapes philippinarum to ocean acidification following gonadal maturation.

Ocean acidification (OA) can seriously affect marine bivalves at different levels of biological organization, generating widespread consequences on progeny recruitment and population maintenance. Yet, few effort has been devoted to elucidating whether female and male bivalves respond differentially to OA in their reproductive seasons. Here, we estimated differences in physiological responses of female and male Manila clams (Ruditapes philippinarum) to OA during gonadal maturation. In comparison to OA-stressed male clams, females significantly depressed activities in enzymes related to energy metabolism (NKA, T-ATP), antioxidant defence (SOD and MDA), and non-specific immune function (ACP), and downregulated expression of AMPK that plays a key role in cellular metabolism, indicating that sex did significantly affect responses of R. philippinarum to OA. Such sex-based differences can be likely couched in energetic terms, given the much more energetically expensive cost of egg production than that of sperms. These results indicate that sex-specific responses to OA during reproductive seasons do exist in marine bivalves, and therefore accounting for such sex specificity is of paramount importance when projecting population sustainability and formulating conservation strategies in an acidifying ocean.

Comparative transcriptome analysis of different tissues of Solanum khasianum reveals candidate genes involved in steroidal glycoalkaloid biosynthesis.

Fruits and leaves of Solanum khasianum C. B. Clarke have long been used as a common Chinese herbal medicine. Steroidal glycoalkaloids (SGAs), the main active ingredient in S. khasianum, exhibit various pharmacological effects. However, genes involved in the SGA biosynthetic pathway in S. khasianum have not yet been identified. Genes encoding potential key SGA biosynthesis enzymes were identified through comprehensive RNA sequencing analysis (RNA-seq) of S. khasianum leaves, stems, and fruits. A total of 123,704 unigenes were obtained, of which 109,775 (88.74%) were annotated in seven public databases. Among these, 54 unigenes potentially involved in SGA biosynthesis were identified. Additionally, 23,636 differentially expressed genes were identified by comparing gene expression levels among the fruits, stems, and leaves of S. khasianum. The structural characteristics and phylogenetic relationship of cycloartenol synthase involved in SGA biosynthesis were further analyzed. Solasodine constituent was detected by high-performance liquid chromatography. This is the first study to report the comparative transcriptome analysis of different tissues of S. khasianum that identifies valuable genes potentially involved in SGA biosynthesis in this species.

Physiological and biochemical responses of clams to recurrent marine heatwaves.

In the past decade, the frequency, intensity and duration of marine heatwaves (MHWs) in the South China Sea have been increasing strikingly, resulting in serious impacts on intertidal bivalves and their ecosystems. The Manila clam, Ruditapes philippinarum, is one of the most ecologically and economically important bivalve species in the South China Sea, yet very little is known about its fate under intensifying MHWs events. Here, we examined how R. philippinarum responded to two consecutive scenarios of MHWs, with each composed of 4 °C and 8 °C rises of seawater temperatures, respectively. Up to 87% of Manila clams survived recurrent MHWs events, and significant increases in standard metabolic rate occurred predominantly under extreme conditions (+8 °C), indicating that the clams could trigger compensatory mechanisms to mitigate MHWs-induced thermal stress. Following acute and repeated exposures to MHWs, Manila clams showed similar responses in enzymes underpinning energy metabolism (NKA, CMA, and T-ATP), antioxidant defence (SOD, CAT, and MDA), and biomineralization (AKP and ACP), most of which exhibited significantly increasing and then decreasing trends with the intensification of MHWs. Of eight genes associated with physiological tolerance and fitness, ATAD3A, PFK, SOD, and C3 were significantly down-regulated in response to recurrent MHWs events, demonstrating the certain resistance to MHWs. These findings provide a better understanding that marine bivalves hold the potential to acclimate simulated MHWs events from the physiological and molecular processes.

Volatile organic compounds for early detection of prostate cancer from urine.

Prostate cancer (PCa) is one of the most common cancers in men worldwide. Early diagnosis of PCa is extremely challenging due to the lack of effective diagnostic methods. The study presented here aims to evaluate whether urine volatile organic compounds (VOCs) can be used as an emerging diagnostic biomarker for PCa. Gas chromatography-ion mobility spectrometry (GC-IMS) was used to detect VOCs in urine samples from 66 patients with PCa and to comparatively analyze samples from 87 patients with non-cancerous controls (NCs). A total of 86 substance peak heights were detected in urine samples from all patients. Analysis using four machine learning algorithms suggested that the diagnosis of PCa could be effectively facilitated. Ultimately, diagnostic models were constructed based on the four VOCs selected. The AUC for the RF and SVM model were 0.955 and 0.981, respectively. Both the NN and DT diagnostic models also achieved an AUC of 0.8 or more, but their sensitivity or specificity was poor compared to the RF and SVM models.

High-resolution history of oxygen depletion in the SW Baltic Sea since the mid-19th century as revealed by bivalve shells.

The Baltic Sea serves as a model region to study processes leading to oxygen depletion. Reconstructing past low-oxygen occurrences, specifically hypoxia, is crucial to understand current ecological disturbances and developing future mitigation strategies. The history of dissolved oxygen (DO) concentration in some Baltic Sea basins has been investigated in previous studies, but temporally well-constrained, inter-annual and better resolved DO reconstructions are still scarce. Here, we present precisely dated, high-resolution DO record since the mid-19th century reconstructed from Mn/Cashell values of Arctica islandica (Bivalvia) collected in the Mecklenburg Bight. According to the data, this area experienced similar low oxygenation during the second half of the 19th century and the late 20th century, but DO variability increased: A 12-15-yr oscillation prevailed in the 19th century, but a 4-6-year period dominated in the late 20th century. Shortly after the onset of the Industrial Revolution around 1850, Mn/Cashell values increased, indicating a DO decrease, probably caused by strong anthropogenic nutrient input. More recently, phosphate levels and inflows of oxygen-rich North Sea water have been identified as major factors controlling the bottom water oxygenation. For example, the increase in DO in the mid-1990s was linked to the decrease in phosphate content and several Major Baltic Inflows. The strong Ba/Cashell rise between the 1860s and the turn of the century most likely reflects changes in diatom community structure rather than a bloom of mass phytoplankton. This is supported by largely unchanged Mn/Cashell and shell growth. Decadal and multi-decadal cycles of shell growth rate correlated strongly with the Atlantic Multidecadal Variability, likely reflecting changes in atmospheric circulation patterns, precipitation rate and riverine nutrient supply. To further improve the management and protection of ecosystems in the Baltic Sea, a larger number of such high-resolution retrospective studies covering long periods of time and large regions are needed.

Survival and physiological energetics of highly invasive mussels exposed to heatwaves.

The South American mussel, Mytella strigata, is a highly invasive fouling species of great concern along intertidal shores in East and Southeast Asia, posing serious threats to native biodiversity and ecosystems. Intertidal areas, being increasingly attacked by heatwaves over the last decade, are among the most thermally challenging habitats, yet the fate of this highly invasive mussel under scenarios of hotter heatwaves remains unknown. Here, we investigated how M. strigata responded to intensifying heatwaves frequently occurring in the South China Sea. Over 97% of individuals survived the five-day-lasting heatwaves, suggesting their high ability to cope with short-term heatwaves. Virtually unaffected clearance rate and absorption efficiency throughout the course of heatwaves indicate the maintenance of energy acquisition, and significantly decreased respiration rate implies the depression of energy metabolism, generating significant decreases in the O:N ratio when heatwaves occurred. Scope for growth of heatwaves-stressed mussels significantly decreased during initial exposure and then increased over time. These findings indicate the remarkable ability of M. strigata to cope with heatwaves recorded in its invasive habitats and call the attention for the rapid spread of this highly invasive fouling species in the context of climate change.

Ocean acidification stunts molluscan growth at CO2 seeps.

Ocean acidification can severely affect bivalve molluscs, especially their shell calcification. Assessing the fate of this vulnerable group in a rapidly acidifying ocean is therefore a pressing challenge. Volcanic CO2 seeps are natural analogues of future ocean conditions that offer unique insights into the scope of marine bivalves to cope with acidification. Here, we used a 2-month reciprocal transplantation of the coastal mussel Septifer bilocularis collected from reference and elevated pCO2 habitats to explore how they calcify and grow at CO2 seeps on the Pacific coast of Japan. We found significant decreases in condition index (an indication of tissue energy reserves) and shell growth of mussels living under elevated pCO2 conditions. These negative responses in their physiological performance under acidified conditions were closely associated with changes in their food sources (shown by changes to the soft tissue δ13C and δ15N ratios) and changes in their calcifying fluid carbonate chemistry (based on shell carbonate isotopic and elemental signatures). The reduced shell growth rate during the transplantation experiment was further supported by shell δ13C records along their incremental growth layers, as well as their smaller shell size despite being of comparable ontogenetic ages (5-7 years old, based on shell δ18O records). Taken together, these findings demonstrate how ocean acidification at CO2 seeps affects mussel growth and reveal that lowered shell growth helps them survive stressful conditions.

Impacts of marine heatwaves on byssus production in highly invasive fouling mussels.

Marine heatwaves (MHWs) are projected to increase in their frequency, intensity, and duration, causing irreversible and catastrophic consequences for intertidal ecosystems around the world. The highly invasive fouling mussel, Arcuatula senhousia, can cause marked habitat alteration by constructing extremely intense byssal mats, devastating the biodiversity of many intertidal systems, yet very little is known about its fate under conditions of more frequent, hotter and longer MHWs. Here, we assessed impacts of two scenarios of MHWs (low-intensity with 4 °C rise of seawater temperature and high-intensity with 8 °C rise, respectively) on the byssal production of A. senhousia. Mussels exposed to low-intensity MHWs did not show any significant differences in the number, length and diameter of byssal threads, compared with those not thermally stressed. Under high-intensity scenario, the byssus production was significantly depressed, and byssal threads became fewer, shorter and finer, in line with significant decreases in cumulative length and volume. These findings provide a better understanding of responses of invasive fouling mussels such as A. senhousia to MHWs and make a leap forward in linking climate change and biological fouling in marine ecosystems.

Responses of digestive metabolism to marine heatwaves in pearl oysters.

Marine heatwaves (MHWs) have increased in intensity and frequency in global oceans, causing deleterious effects on many marine organisms and ecosystems they support. Bivalves are among the most vulnerable taxonomic groups to intensifying MHWs, yet little is known about the underlying mechanisms. Here, we investigated the impact of MHWs on the digestive metabolism of pearl oysters (Pinctada maxima). Two moderate and severe scenarios of MHWs were performed by increasing seawater temperature respectively from 24 °C to 28 °C and 32 °C for 3 days. When subjected to MHWs and with increasing intensity, pearl oysters significantly enhanced their digestive enzymatic activities, such as lipase and amylase. LC-MS-based metabolomics revealed negative responses in the lipid metabolism (e.g., steroid biosynthesis, glycerophospholipid metabolism, and sphingolipid metabolism), the amino acid metabolism (e.g., glutamate, histidine, arginine, and proline), and the B-vitamins metabolism. These findings indicate that the digestive metabolism of marine bivalves can likely succumb to intensifying MHWs events.

Transcriptomic responses reveal impaired physiological performance of the pearl oyster following repeated exposure to marine heatwaves.

Marine heatwaves are predicted to become more intense and frequent in the future, possibly threatening the survival of marine organisms and devastating their communities. While recent evidence reveals the adaptability of marine organisms to heatwaves, substantially overlooked is whether they can also adjust to repeated heatwave exposure, which can occur in nature. By analysing transcriptome, we examined the fitness and recoverability of the pearl oyster (Pinctada maxima) after two consecutive heatwaves (24 °C to 32 °C for 3 days; recovery at 24 °C for 4 days). In the first heatwave, 331 differentially expressed genes (DEGs) were found, such as AGE-RAGE, MAPK, JAK-STAT, FoxO and mTOR. Despite the recovery after the first heatwave, 2511 DEGs related to energy metabolism, body defence, cell proliferation and biomineralization were found, where 1655 of them were downregulated, suggesting a strong negative response to the second heatwave. Our findings imply that some marine organisms can indeed tolerate heatwaves by boosting energy metabolism to support molecular defence, cell proliferation and biomineralization, but this capacity can be overwhelmed by repeated exposure to heatwaves. Since recurrence of heatwaves within a short period of time is predicted to be more prevalent in the future, the functioning of marine ecosystems would be disrupted if marine organisms fail to accommodate repeated extreme thermal stress.

A high-throughput catalyst synthesis system for Ag-based catalysts.

Ag-based catalysts have been used in many practical reactions, such as p-nitrophenol reduction, due to the advantages of low cost and excellent activity. In order to facilitate the development of Ag-based catalysts, it may be helpful to use automated equipment for experiments. In this study, a system for the high-throughput synthesis of Ag-based catalysts was developed based on a facile impregnation method. Notably, the system automates the batch synthesis of Ag-based catalysts by setting the catalyst formulation in a dedicated software. Moreover, the software used employs the ant colony algorithm to optimize the synthesis path and improve the synthesis efficiency. The catalysts obtained from the high-throughput system are found to be similar to the manually prepared samples based on comparison of characterization results. In addition, experiments also reveal that this high-throughput system is capable of achieving high-throughput synthesis of Ag-based catalysts at the gram level. The synthesis of Pt-Ag bimetallic catalysts shows that this high-throughput system can be effectively used for exploratory experiments. This work paves the way for a high-throughput technique to synthesize Ag-based catalysts in a short period of time, which could be extended to the preparation of other catalyst systems. Moreover, the high-throughput synthesis system of Ag-based catalysts provides a feasible prerequisite for subsequent high-throughput characterization, which is a significant advancement in the development of industrial catalysts.

Post-responses of intertidal bivalves to recurrent heatwaves.

Heatwaves are becoming hotter, longer and more frequent, threatening the survival of intertidal bivalves and devastating their ecosystems. Yet, substantially overlooked are heatwave-induced post-responses, which are important to assess cascading consequences. Here, we investigated responses of intertidal bivalves, Ruditapes philippinarum, to recurrent heatwaves. Physiological and gene expression analyses demonstrated that the mantle tissue of R. philippinarum did not sensitively respond to heatwaves, but revealed post-responses under recovery scenarios. Of 20 genes related to essential physiology and fitness, 18 were down-regulated during the 1st recovery period, but following repeated exposure, 13 genes were up-regulated, in line with significantly increased activities of energy-metabolizing enzymes, and antioxidant and nonspecific enzymes. The down-regulation of genes involved in biomineralization, nevertheless, was observed under recovery scenarios, implying the trade-off between essential physiological and fitness-related functions. These findings pave the way for understanding the physiological plasticity of marine bivalves in response to intensifying heatwaves.