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This is the first example of the construction of harmaline-based spiropolycyclic scaffolds using harmaline-based N–C synthons and evaluation of 3cb as an anticancer agent.

The first example of 3-vinyl oxindole-chromones in the ring opening and recyclization reaction.

Herein is reported the first example of ring opening and skeletal reconstruction of 3-vinyl benzofuranone-chromones 1 as versatile synthons, which can react with ammonia or primary aliphatic amines as binucleophiles, for the eco-friendly and atom-economical synthesis of diverse and functionalized 2-pyridones 3 with potential biological activity in good to excellent yields (77-93%). When using optically active 1,2-diphenylethylenediamine 2 as the binucleophile, the

目的：透過異常通報資料庫分析，探討96小時對照72小時重置周邊靜脈導管，對於靜脈炎發生率之影響。 方法：方便取樣臺灣南部某醫學中心之異常通報資料庫，分析2011年9月1日至2014年8月31日之72小時重置組，與2014年9月1日至2017年8月31日之96小時重置組，2個時期之住院個案的靜脈炎發生率。 結果：96小時對照72小時重置周邊靜脈導管，並未增加靜脈炎發生率。靜脈炎總發生率（勝算比[odds ratio,OR] = 0.70, p=.0290）與細菌性靜脈炎發生率(OR= 0.11, p=.0097)顯著減少，化學性與機械性靜脈炎發生率則無顯著差異。 結論：本土性資料庫分析結果為在臺灣熱帶海島型潮濕氣候，三班評估無靜脈炎症狀下，96小時重置周邊靜脈導管並不會顯著增加靜脈炎風險，建議醫療機構可據此調整臨床作業規範，落實實證知識轉譯。 "Purpose: To compare the effect of replacement of peripheral venous catheters at 96-hour intervals on the incidence of phlebitis with that at 72-hour intervals through analysis of an incident-reporting database. Methods: Convenience sampling of records from the incident-reporting database of a medical center in southern Taiwan was used to analyze the incidence of phlebitis among hospitalized patients with replacement of peripheral venous catheters at 72-hour intervals (from September 1, 2011, to August 31, 2014) or 96-hour intervals (from September 1, 2014, to August 31, 2017). Result: The analysis revealed that replacing peripheral venous catheters every 96 hours rather than every 72 hours did not increase the incidence of phlebitis. The total incidence of phlebitis (odds ratio [OR]=0.70, p=.0290) and the incidence of bacterial phlebitis (OR=0.11, p=.0097) decreased significantly, and no significant differences in the incidence rates of chemical or mechanical phlebitis were identified. Conclusion: According to the analysis results of a local database, under the humid tropical insular climate of Taiwan, replacement of peripheral venous catheters at 96-hour intervals did not significantly increase the risk of phlebitis in patients who exhibited no symptoms of phlebitis as assessed in three shifts. Medical institutions can adjust their clinical operation standards and implement knowledge translation accordingly. &nbsp

Maintaining the balance of intestinal microbiota is vital for the growth and health of crustaceans. The current study explored the influences of the surface layer proteins (SLPs) of Pediococcus pentosaceus on the growth performance, disease resistance, and gut microbial community of the freshwater prawn Macrobrachium rosenbergii. After 28 days of feeding, obvious enhancements in the growth performance were found in the PF (a commercial basic diet + intact P. pentosaceus) group compared to the CF (a commercial basic diet, control) and RF (a commercial basic diet + LiCl-treated P. pentosaceus) groups. Meanwhile, the relative percent survival (RPS) of the PF group was found to be 82.1%, higher than the RF group (RPS = 63.2%) in the challenge test. There were noteworthy differences in the relative richness of Aeromonas among different treatments, and the PF group was the lowest. Whereas the relative abundance of Lactococcus between the CF and RF group did not markedly differences, which was obviously lower than the PF group. The prediction of bacterial phenotype indicated that the pathogenic potential of intestinal microbiota was decreased by increasing dietary SLPs-carrying P. pentosaceus. The results suggested that SLPs-carrying P. pentosaceus can improve growth, modulate intestinal microbiota, and increase the resistance of prawns against Aeromonas hydrophila infection.

Spinel-structured lithium nickel manganese oxide (LiNi0.5Mn1.5O4, LNMO) is a low-cost battery cathode material with a high operating voltage (∼4.7 V vs Li+/Li), relatively high capacity (∼147 mA h ...

p70S6K is involved in cellular response, such as tumor metastases, the immune response and tissue repair in vertebrates. The role of p70S6K in these physiological processes in crustaceans remains, however, unknown. In this study, the Lvp70S6K was identified, containing a 5' UTR of 294 bp, an ORF of 1494 bp ad a 3' UTR of 211 bp, encoding 497 amino acids with a theoretical molecular weight of 70 kDa and an estimated isoelectric point of (pI) of 5.16. The multiple alignment found that Lvp70S6K was highly homologous with other invertebrates. Lvp70S6K mRNA was detected in all the tested tissues and the Lvp70S6K expression levels was significantly down-regulated and reached the lowest level (0.44-fold, p 0.01) at 1.5 h after low temperature stress. The subcellular localization of Lvp70S6K could be detected in cytoplasm. ROS production was significantly up-regulation (1.19-fold, p 0.01), total hemocyte count (THC) was significantly down-regulation (0.22-fold, p 0.01), apoptosis rate was markedly increased (1.09-fold, p 0.01), apoptosis-related genes of LvPDCD4 (1.61-fold, p 0.01) and LvCyt.C (1.23-fold, p 0.01) were up-regulated, and anti-apoptotic gene of LvBcl-2 (0.69-fold, p 0.01), LvIAP1 (0.68-fold, p 0.01) and LvIAP2 (0.45-fold, p 0.01) were decreased after low temperature stress in hemolymph of Lvp70S6K-silenced shrimp at 1.5 h. Silencing of LvPTEN significantly increased Lvp70S6K, LvPI3K, LvAKT and LvmTOR expression. In summary, these results indicated that Lvp70S6K play a crucial role in oxidative and apoptosis, which was able to negatively regulate by PTEN.

In this paper, the structure and magnetic behaviour of antiperovskite InxFe4-xN have been investigated systematically. The crystal lattice becomes larger and the Curie temperature decreases with increasing x. The magnetic state changes from a ferromagnetic to a glassy state. In addition, an obvious spin glass (SG) behaviour has been revealed in In0.6Fe3.4N (x = 0.6) with a freezing temperature of T0 = 73 K, dynamical exponent of zν = 5.51, and flipping time of τ0 = 4.26 × 10−11 s. The origin of the SG behaviour in In0.6Fe3.4N may be attributed to atomic disorders at Wyckoff position 1a or ferromagnetic frustrations, or the both.

As a new antiperovskite nitride, ZnFe3N was synthesized and characterized by almost completely substituting iron atoms at corner positions of γ'-Fe4N. The magnetic interactions of the system with the space group Pm3[combining macron]m are fully investigated. The critical behavior was investigated based on the measured magnetic data around the ferromagnetic phase transition temperature. In this work, the values of critical exponents (β, γ and δ) were obtained systematically using the Kouvel-Fisher method in the critical region. The Widom scaling law (δ = 1 + γβ-1) and the scaling equation (m = f±(h)) were used to reveal the reliability of these values. The values of the critical exponents (β = 0.325, γ = 1.228, and δ = 4.778) are different from those predicted by the three-dimensional (3D) Heisenberg model and mean-field model, and are very close to those of the 3D-Ising model. Combined with ESR analysis, the spin clusters induced by changes in chemical bonds are considered to be the cause for the existence of an anisotropic short-range ordered state in this ferromagnetic system.

Background Primary malignant melanoma of the esophagus accounts for 0.1%-0.2% of all esophageal malignancies, including melanotic and amelanotic melanomas. Primary amelanotic malignant melanoma of the esophagus is extremely rare, and only about 20 cases have been published in the literature to date. Most primary malignant melanomas of the esophagus are diagnosed following development of metastatic lesions and thus have a very poor prognosis. The median survival duration of patients with metastatic melanoma has been reported to be 6.2 mo. Case summary A 49-year-old woman was referred to our hospital with a diagnosis of esophageal cancer. Endoscopy, biopsy, imaging evaluation, and physical examination at our hospital indicated a diagnosis of advanced primary amelanotic malignant melanoma of the esophagus. Immunohistochemical staining confirmed melanoma. Nuclear medicine examination revealed a left iliac bone metastatic lesion. After discharge, the patient self-administered apatinib for 3 mo, followed by oral treatment with Chinese medicines (also self-administered) for 2 mo. No treatments had been taken since then. The patient has survived with no growth out to the most recent follow-up (24 mo post diagnosis), and she always presented with a positive attitude about her condition during this period. Conclusion Survival following metastatic melanoma might be related to the pharmaceutical and Chinese medicine treatment and the patient's positive attitude.

Organozinc reagents are among the most commonly used organometallic reagents in modern synthetic chemistry, and multifunctionalized organozinc reagents can be synthesized from structurally simple, readily available ones by means of alkyne carbozincation. However, this method suffers from poor tolerance for terminal alkynes, and transformation of the newly introduced organic groups is difficult, which limits its applications. Herein, we report a method for vinylzincation of terminal alkynes catalyzed by newly developed iron catalysts bearing 1,10-phenanthroline-imine ligands. This method provides efficient access to novel organozinc reagents with a diverse array of structures and functional groups from readily available vinylzinc reagents and terminal alkynes. The method features excellent functional group tolerance (tolerated functional groups include amino, amide, cyano, ester, hydroxyl, sulfonyl, acetal, phosphono, pyridyl), a good substrate scope (suitable terminal alkynes include aryl, alkenyl, and alkyl acetylenes bearing various functional groups), and high chemoselectivity, regioselectivity, and stereoselectivity. The method could significantly improve the synthetic efficiency of various important bioactive molecules, including vitamin A. Mechanistic studies indicate that the new iron-1,10-phenanthroline-imine catalysts developed in this study have an extremely crowded reaction pocket, which promotes efficient transfer of the vinyl group to the alkynes, disfavors substitution reactions between the zinc reagent and the terminal C-H bond of the alkynes, and prevents the further reactions of the products. Our findings show that iron catalysts can be superior to other metal catalysts in terms of activity, chemoselectivity, regioselectivity, and stereoselectivity when suitable ligands are used.

Background The tumor microenvironment (TME) has received an increasing amount of attention. CXC chemokines can regulate immune cell transport and tumor cell activity to exert anti-tumor immunity. However, studies on the expression and prognosis of CXC chemokines in cervical cancer (CC) are more limited. Methods The study investigated the role of CXC chemokines in TME of CC by using public databases. Moreover, quantitative real-time PCR (qRT-PCR) and immunohistochemistry (IHC) of CXC chemokines were performed to further verify. Results The transcriptional levels of CXCL1/3/5/6/8/9/10/11/13/16/17 in CC tissues were significantly elevated while the transcriptional levels of CXCL12/14 were significantly reduced. We reached a consistent conclusion that the expression of CXCL9/10/11/13 was verified by quantitative real-time PCR and immunohistochemistry. Moreover, CC patients with low transcriptional levels of CXCL1/2/3/4/5/8 were significantly associated with longer overall survival (OS). The CCL family was related to CXC chemokines neighboring alteration. RELA, NFKB1, LCK and PAK2 were the key transcription factors and kinase targets of CXC chemokines, respectively. We also found there were significant correlations between the expression of CXCL9/10/11 and the infiltration of immune cells (CD8+ T cell, CD4+ T cell, neutrophils and dendritic cells). Conclusions In brief, we conducted a comprehensive analysis of CXC chemokines via clinical data and some online public databases. Our results may provide a new idea for the selection of immunotherapeutic targets and prognostic biomarkers for cervical cancer.

MicroRNAs (miRNAs) play key roles in many physiologic and pathologic processes, including autophagy. Autophagy is cellular in an emergency response mechanism of environment stress, but their complex molecular regulatory mechanism under low-temperature stress is largely unknown in shrimp, especially miRNA-mediated regulation of autophagy in low-temperature tolerance. In this article, a shrimp PvTOR and miRNA pva-miR-151 cooperation in response to low-temperature stress has been reported. Pva-miR-151 showed expression patterns opposite to target PvTOR under low-temperature stress. The pva-miR-151 targets the 3′-UTR region of PvTOR, regulate the formation of autophagosome, which contribute to the degradation and recycling of damaged organelles. In addition, the low-temperature tolerance was correlated positively with autophagy in shrimp. Silenced pva-miR-151 increased sensitivity to low-temperature stress, whereas overexpression pva-miR-151 decreased the expression of PvTOR and p-TOR and increased tolerance to low-temperature stress by improving the formation of autophagosome and total hemocyte count. In addition, the TOR activator 3BDO can partially rescue autophagy induced by overexpression of pva-miR-151; these results indicate that miR-151 was necessary for the low-temperature tolerance in shrimp. Taken together, we provide a novel strategy and mechanism for shrimp breeding to improve shrimp low-temperature tolerance.

Water temperature is one of the most common physiological stressors in aquaculture. Previous studies demonstrate that organisms require miRNA activity for survival in various unfavourable environmental conditions. However, the detailed role of miRNA in response to low-temperature stress is still unclear. This study was conducted to construct a comprehensive miRNA dataset for the Penaeus vannamei after low-temperature stress. A total of 329 known miRNAs and 60 putative novel miRNAs were identified. Among them, 17 miRNAs were identified with the most significant differences, and they were found to be involved in stimulation or stress processes. The main enriched target pathways of the 17 miRNAs were the Hippo signalling pathway, autophagy, apoptosis and MAPK signalling. In addition, all the 17 miRNAs identified were up-regulated, suggesting that miRNA by inhibiting the expression of target genes constitutes an effective strategy for Penaeus vannamei to cope with low-temperature stress. The 35-putative target of the 17 miRNAs was related to apoptosis and autophagy-related proteins, such as Pxt, DRAM2, cytochrome c, ATG2B, JNK, ATG4 and API5. The analysis of miRNA expression profiles contributes to the understanding of the molecular mechanisms of low-temperature tolerance in Penaeus vannamei. This study's findings enrich current miRNA resources and offer the possibility to validate the involvement of 17 miRNAs in the response of shrimp to low-temperature stress.

Proline (Pro) metabolism is intimately associated with stress adaptation. The catabolism of Pro includes two dehydrogenation reactions catalyzed by proline dehydrogenase (ProDH) and Δ1-pyrroline-5-carboxylate dehydrogenase (P5CDh). P5CDh is a mitochondrial matrix NAD+-dependent dehydrogenase that is critical in preventing P5C-Pro intensive cycling and avoiding ROS production from electron run-off. Little is known about the roles of P5CDh in invertebrates, however. We cloned the P5CDh sequence in the Pacific white shrimp, Litopenaeus vannamei, and found that LvP5CDh is expressed predominantly in pleopod, hepatopancreas and gill. Subcellular localization analysis revealed that LvP5CDh protein was mainly found in the cytoplasm. In addition, overexpressing LvP5CDh in cells reduced ROS formation and inhibited apoptosis induced by LC50 Cd2+. Shrimp were exposed to various stress factors including infection with Vibrio alginolyticus, (½ LC50 and LC50) Cd2+, acid (pH 5.6) and alkali stress (pH 9.3). Both biotic and abiotic stress resulted in increased LvP5CDh expression and Pro accumulation; V. alginolyticus infection, pH 9.3 and LC50 Cd2+ stress apparently stimulated the Glu pathway of Pro synthesis, while pH 5.6 and ½ LC50 Cd2+ stress promoted the Orn pathway of Pro synthesis. Silencing of Lvp53 in shrimp attenuated LvP5CDh expression during Cd2+ stress, but had no effect on LvP5CDh mRNA levels if no Cd2+ stress was imposed. Our study contributes to the functional characterization of LvP5CDh in biotic and abiotic stress and reveals it to protect against ROS generation, damage to the cell, including the mitochondria, and apoptosis. Thus, LvP5CDh plays a critical role in immune defense and antioxidant responses.

Nucleotide excision repair (NER) removes many different types of DNA lesions, and NER related host factors are reported to aid recovery steps during viral integration. Here, we report the identification and characterization of a DNA repair gene Rad23 from Litopenaeus vannamei and explore its role in innate immunity of crustaceans. LvRad23 contains a1149 bp open reading frame (ORF) which encodes a 382 amino acids protein with predicted theoretical isoelectric point of 4.21. LvRad23 was ubiquitously expressed in the muscle, eyestalk, gill, stomach, heart, legs, intestine, and hepatopancreas in order from high to low and LvRad23 protein was showed to be located in the cytoplasm of Drosophila S2 cells. The homology analysis showed that it has a high sequence homology with Rad23 protein from Marsupenaeus japonicus. Vibrio alginolyticus challenge induced a remarkable up-regulation of LvRad23 mRNA in hepatopancreas. Knocking down LvRad23can interfere the NER pathway by down regulating the expression of replication protein A (RPA) and proliferating cell nuclear antigen (PCNA). However it didn't cause any significant difference on total hemocyte count (THC) between LvRad23-silenced and non-silenced group.LvRad23-silenced then challenge with V. alginolyticus inducing high level of reactive oxygen species (ROS) and DNA damage in hemolymph. As well as decreased THC, which seriously diminished the innate immune system of L. vannamei. Meanwhile, the NER pathway was reactived by enhancing the expression of LvRad23 and promoting the production of LvPCNA to resist apoptosis and maintain proliferation of hemolymph cells in the later stage. Our results suggest that LvRad23 plays a vital role in shrimp specific immune response to V. alginolytcus through its participation in NER pathway.

Clofibric acid (CFA), a drug and personal care product, has been identified as ubiquitous in the aquatic system and surface water, causing pollution to the environment. In this study, after environmental (4 µg/L) levels of CFA challenge, the LvFABP, LvACS gene expressions, total haemocyte count (THC), relative enzymes (SOD1 and GST) activities in Litopenaeus vannamei were observed to decrease. In the meantime LvFATP, LvRXR expression and the level of NEFA were upregulated in L. vannamei body. LvFABP expression in vivo was knocked down by dsRNA-mediated RNA interference (RNAi), which led to significantly decreased levels of PPARα (including LvFATP, LvRXR and LvACS). When exposed to environmental CFA after 4 days, LvFABP knocked down group had a sharp upregulation of LvFATP, LvRXR, LvACS expression, GST activity and NEFA amount, following decreased THC and SOD1 activity. These results suggested that environmental concentration CFA may have some toxicological effect on L. vannamei, following fatty acids metabolism and oxidative stress responses by LvFABP via the PPARα/RXR signaling pathway, including LvFATP, LvRXR and LvACS.

As a crucial molecular switch, Cdc42 is a signal regulation hub which is involved in a wide range of cellular processes, including cytokinesis, gene expression, cell cycle progression and apoptosis. It has been reported that this GTPase promotes host defense against fatal infection and plays a vital role in the innate immunity system of mammals. But whether and how Cdc42 participates in innate immunity in invertebrates, such as the shrimp Litopenaeus vannamei , is still unknown. In this study, confocal microscopy analysis showed that LvCdc42 located in both cytoplasm and nucleus of S2 cells depended on its structure. The silencing LvCdc42 induced an increase in the expression of Lvp53 and Lvcaspase-3. When LvCdc42-silenced shrimps were stressed with Vibrio alginolyticus , the expression of Lvp53 and Lvcaspase-3 was markedly up-regulated. Moreover, the increase in the apoptosis rate in hemocytes and in cumulative mortality were in line with Lvp53 mRNA expression. These data suggest that the molecular switch LvCdc42 acts as a negative regulator of Lvp53 and participates in the apoptosis of hemocytes when L. vannamei is challenged with V. alginolyticus .

Ficus hirta Vahl. has been reported to have hepatoprotective, antitumor, antibacterial functions, and is used to treat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Ammonia nitrogen is one of the most common environmental stress factors in aquaculture. Long-term exposure to high concentrations of ammonia nitrogen can induce oxidative stress and increase the risk of infections. However, whether Ficus hirta Vahl. has effect on ammonia nitrogen stress is unclear. In present study we report that Ficus hirta Vahl. improves the activity of total antioxidant capacity (T-AOC), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) of shrimp and decreases shrimp mortality caused by ammonia nitrogen stress. It is demonstrated that miR-2765 is negatively regulate the antioxidant capacity. We find that SOD was a direct target gene of miR-2765. MiR-2765 can bind to 3'-untranslated region (3'-UTR) of SOD to inhibit its transcription. Furthermore, Ficus hirta Vahl. down-regulates miR-2765 to activate the antioxidant capacity to alleviate the damage caused by ammonia nitrogen stress. Interestingly, overexpression of miR-2765 could attenuate the protective effect of Ficus hirta Vahl. on shrimp under ammonia nitrogen stress. These data indicate that Ficus hirta Vahl. alleviates the damage of ammonia nitrogen stress in shrimp by repressing miR-2765 and activating the antioxidant enzyme system. This study will provide a theoretical basis and a new perspective for assessing the toxicity mechanism of ammonia nitrogen in the process of farming on shrimp.

Smad3 is a key mediator of the canonical TGF-β signaling pathway and plays an important role in TGF-β1-mediated transcriptional regulation. However, the function of Smad3 in crustaceans such as shrimp, is still poorly understood and needs to be further explored. We characterized Litopenaeus vannamei Smad3 (LvSmad3) and its biological functions were investigated in response low temperature stress. Full-length LvSmad3 cDNA was 2341bp and contained an open reading frame (ORF) of 1326 bp that encoded a 441 amino acid long protein, with a predicted molecular mass of 48.35 kDa. Phylogenetic analysis revealed that LvSmad3 has a high degree of similarity with other known species. LvSmad3 mRNA was detected in all the tested tissues and highest transcription occurred mostly in gills. Further research showed that suppressing the expression of Smad3 could reduce ROS production, DNA damage and the apoptosis rate in shrimp hemocyte under low temperature compared with the dsGFP group. Thus, we speculated that Smad3 could promote the apoptosis of hemocytes. We confirmed that Smad3 could inhibit apoptosis in the hepatopancreas by suppressing the expression of pro-apoptotic genes. Taken together, the silencing of Smad3 can reduce ROS production induced by low temperature stress, weaken the damage to hemocytes and the hepatopancreas by inhibit the apoptosis.

MicroRNAs (miRNAs) are critical post-transcriptional regulators, which play a crucial role in resistance to adverse environmental stress by regulating autophagy. However, the mechanism of miRNA involved in the autophagy regulation of shrimp under ammonia nitrogen stress is still limited. In the present study, ammonia nitrogen could induce hepatopancreas injury and oxidative stress of P. vannamei, and significantly increase the content of ROS in hemocytes by flow cytometry. Simultaneously, it is accompanied by autophagy occurred in the hemocytes and hepatopancreas. Furthermore, the qRT-PCR analysis revealed that the expression of pva-miR-252 in P. vannamei decreased significantly after ammonia nitrogen stress, and pva-miR-252 negatively regulated PvPI3K by binding to 3'UTR of PvPI3K by double-luciferase assay. Pva-miR-252 overexpression could significantly increase the level of autophagy, and restore the autophagy inhibition caused by Chloroquine in vitro , whereas silencing of pva-miR-252 resulted in the opposite effect. More importantly, overexpression of pva-miR-252 could enhance the activity of antioxidant enzymes and reduced the production of ROS of shrimp under ammonia nitrogen stress. In conclusion, pva-miR-252 could positively regulate autophagy through PvPI3K and improve the antioxidant enzyme activity of P. vannamei under ammonia nitrogen stress, and our study provides a novel theoretical molecular mechanism for further understanding the shrimp cope with a high ammonia nitrogen environment.

Bcl-2 is a pro-survival member of Bcl-2 like superfamily, playing an important role in regulating the apoptotic process. In this study, the full-length Bcl-2 (EcBcl-2) was obtained, consisting of a 5′UTR of 290 bp, an ORF of 699 bp and a 3′UTR of 920 bp. EcBcl-2 gene encoded a polypeptide of 232 amino acids with an estimated molecular mass of 26.12 KDa and a predicted isoelectric point (pI) of 6.93. The deduced amino acid sequence analysis showed that EcBcl-2 consisted of the conserved residues and characteristic domains known to the critical functionality for Bcl-2. qRT-PCR analysis revealed that EcBcl-2 transcript was expressed in all the examined tissues, while the strongest expression level was observed in liver, followed by the expression in blood, gill, kidney, spleen, heart, intestine and muscle. The groupers challenged with V. alginolyticus showed a significant increase of EcBcl-2 mRNA in immune tissues. In addition, western blotting analysis confirmed that the up-regulation of EcBcl-2 protein expression was detected in liver. Subcellular localization analysis revealed that EcBcl-2 was localized in both nucleus and cytoplasm. Overexpression of EcBcl-2 can inhibit the LPS-induced apoptosis and activate the transcription activity of NF-κB and AP-1, while the deletion of BH1, BH2, BH3 or BH4 domain from EcBcl-2 can impede the signaling transduction. These results indicate that EcBcl-2 may play a regulatory role in the apoptotic process.

Incorporation of foreign elements into the cathode material is broadly adopted by both academia and industry to improve the battery performance. The lack of an in-depth understanding for the underlying mechanism, however, makes it a largely try-and-error process with unsatisfactory efficiency and effectiveness. This is particularly true for the electrochemical reaction kinetics that is heterogeneous over a broad range of length scales and is determined collectively by the cathode’s electronic structure, lattice configuration, and micro-morphology. Here we unveiled a facet-dependent dopant segregation effect in Zr-modified single-crystal LiNi0.6Co0.2Mn0.2O2 cathode. By forming kinetically favored corners on the cathode particles, the presence of a trace amount of Zr critically modulates the mesoscale reaction kinetics. Our findings suggest that a delicately controlled dopant distribution is a viable strategy for designing the next-generation battery cathode with superior structural and chemical robustness.

The present study investigated the role of echinocystic acid (EA) on the expression of nuclear factor (NF)-κB and cytochrome P450 1A1 (CYP1A1), and aortic morphology, in a rat model of hyperhomocysteinemia (Hhcy). A total of 50 Sprague Dawley rats were randomly divided into five groups as follows: Normal control (NC), model control (MC), vitamin control (VC; folic acid 1 mg/kg + vitamin B2 2 mg/kg + vitamin B12 10u g/kg), EA1 (20 mg/kg EA) and EA2 (40 mg/kg EA). Plasma homocysteine (Hcy) levels were determined via high performance liquid chromatography, and the morphology of the aorta was investigated using hematoxylin and eosin staining. Furthermore, aortic mRNA and protein levels of NF-κB and CYP1A1 were measured using reverse transcription-quantitative polymerase chain reaction analysis and western blotting, respectively. Plasma Hcy levels, and aortic mRNA and protein levels of NF-κB and CYP1A1, were significantly lower in the EA-treated group compared with the MC group (all P

The serine/threonine protein kinase MST4 plays multiple roles in the regulation of signaling pathways that govern cellular processes including mitosis, migration, homeostasis, polarity, proliferation, differentiation and apoptosis. Here we report the identification and characterization of the full-length sequence of LvMST4 from the shrimp L. vannamei, and investigations into its role in the shrimp's immune response to infection by the pathogenic bacterium Vibrio alginolyticus. Subcellular localization assays demonstrated the enzyme's presence in the shrimp's cytoplasm, and tissue-specific expression analysis revealed that it is expressed ubiquitously but at different levels in different tissues. Infection with V. alginolyticus increased LvMST4 expression and induced a rapid response via the TLR-TRAF6 signaling pathway, causing a decline in the total hemocyte count (THC) and an increase in respiratory burst (RB) activity. In non-infected shrimp, RNAi silencing of LvMST4 with dsRNA had no significant effect on THC but seemed to activate the TRAF6-MKK6-p38 pathway and reduced RB activity. In shrimp challenged with V. alginolyticus, LvMST4 silencing reduced bacterial clearance and increased the initial upregulation of LvTRAF6 while reducing the expression of LvMKK6 and Lvp38. LvMST4 silencing also slightly reduced the THC but caused pronounced increases in RB activity and cumulative mortality. These findings suggest that LvMST4 contributes to antimicrobial responses via the TLR-TRAF6 signal pathway, and helps maintain immunological homeostasis in L. vannamei.

The Pacific white shrimp (Litopenaeus vannamei), as an economically important species, has been reared in low-salinity water during the last decade. To investigate how juvenile L. vannamei shrimp fed with fructose-enriched Artemia respond to acute low-salinity stress, the shrimp were randomly divided into four treatment groups, three groups were fed with Artemia enriched with either 100, 200 and 300 mg L−1 of fructose and a control group fed with Artemia with no enrichment for 10 days. The results showed that the 300 mg L−1 fructose group demonstrated the maximum survival rate and glycogen content. Additionally, the 300 mg L−1 fructose group showed significantly higher Na+/K+-ATPase activity, total antioxidant capacity, and expression levels of Na+/K+-ATPase α-subunit, V-H ATPase α-subunit, hexokinase, phosphofructokinase, antioxidants (SOD, CAT, GPX) and Hsp70 mRNA when compared with the control group. Furthermore, after exposure to low salinity, the mRNA levels of phosphofructokinase, V-H ATPase α-subunit, GPX, p38, JNK and Rac1 stayed constant in shrimp fed with fructose-enriched Artemia but changed significantly in the control group. Thus, a diet of fructose-enriched Artemia can improve the osmoregulation and survival of juvenile L. vannamei shrimp exposed to low salinities.

Target of rapamycin (TOR) is an atypical of Ser/Thr protein kinase that plays an important role in many aspects such as cell growth, reproduction, differentiation, cell cycle regulation, autophagy and apoptosis. However, little information is known about the enzyme in crustaceans. Here, a novel TOR was identified from shrimp Penaeus vannamei (PvTOR) and its biological functions were investigated in response low temperature stress. The PvTOR gene encoded a polypeptide of 2464 amino acids with an estimated molecular mass of 279.4 kD and a predicted isoelectronic point (pI) of 7.30. Phylogenetic analysis revealed that PvTOR shared high similarity with other known species. PvTOR mRNA was detected in all the tested tissues and highest transcription in muscle and hepatopancreas. PvTOR transcriptional level was up-regulated significantly at 1.5 h and 3 h, and down-regulated at 12 h and 24 h after low temperature stress. TEM and autophagy indicator system GFP-PvLC3 suggested that low temperature induced autophagy generation. ROS, Ca2+ concentration and apoptosis rate were increased significantly in TOR-knockdown shrimp after low temperature stress. The autophagy associated gene ATG8II/I, PvBeclin-1, PvATG14, apoptosis gene PvPARP, Pvcasp-3, PvBAX and Pvp53 transcripts, and casp-3/8 activity in hemocyte were increased significantly in TOR-knockdown group shrimp at 3 h after low temperature stress. Additionally, THC counts of TOR-knockdown group were significantly higher than the dsGFP group. In summary, these results suggested that PvTOR plays an important role in the adaptation mechanisms of shrimp at low temperature by regulating autophagy and apoptosis.

MicroRNAs (miRNAs) are a kind of small non-coding RNAs that have been reported to play a vital role in mediating host-pathogen interactions. High-throughput sequencing technology was applied to identify and illuminate mRNAs and miRNAs from grouper infected with Vibrio alginolyticus. The KEGG pathway enrichment analysis showed that the most significate DEGs are associated with Toll-like receptor signaling pathway and NOD-like receptor signaling pathway. We obtained 374 known miRNAs and 116 novel miRNAs. During them, there are 31 up-regulated miRNAs and 93 down-regulated miRNAs. miRNA-mRNA GO and KEGG analysis show that there are 90 miRNAs associated with the immune system. The target genes of immune-related miRNAs (miR-142, miR-146, miR-150, miR-155, miR-203, miR-205, miR-24, miR-31) and genes (CD80, IL-2, AMPK, PI3K) in Epinephelus coioddes were predicted and validated. This study provides an opportunity to further understanding the molecular mechanisms especially the immune system of miRNA regulation in Epinephelus coioddes host-pathogen interactions.

SAHH is an enzyme, playing a significant role in the catalyzation of the S-adenosyl homocysteine (SAH) into homocysteine (Hcy) and adenosine (Ado). However, little is known information of the enzyme in crustaceans. In the present study, SAHH cDNA was cloned from Litopenaeus vannamei (LvSAHH). The full length of the LvSAHH was found, containing a 5' UTR of 119 bp, an ORF of 1236 bp and a 3' UTR of 549 bp. The LvSAHH gene encoded a polypeptide of 411 amino acids with an estimated molecular mass of 45.55 kD and a predicted isoelectronic point (pI) of 5.63. Comparison of the deduced amino acid sequence showed that LvSAHH has high identity (70 %-82%) with other known species. qRT-PCR analysis revealed that LvSAHH mRNA was broadly expressed in all of the examined tissues, while the highest expression level was observed in muscle, followed by the expression in stomach, gill, pleopod, hepatopancreas, heart, eye and intestine. Subcellular localization analysis revealed that LvSAHH was predominantly localized in the cytoplasm and nucleus. LvSAHH mRNA expression levels in hepatopancreas and gill were significantly up-regulated from 6 to 48 h after V. alginolyticus injection and reached the highest level (15-fold and 8-fold, p 0.01) at 24 h, respectively. Additionally, the Toll-like receptors (TLR) and interleukins-16 (IL-16) were detected in hepatopancreas and gill of LvSAHH-knockdown SAHH. LvRack1, LvToll1, LvToll2, LvToll3 and LvIL-16 transcripts were decreased significantly in LvSAHH-knockdown shrimp at 24 h post V. alginolyticus stimulation in hepatopancreas and gill. But LvToll3 was no significant difference in gill. In summary, these results indicated that LvSAHH may play a regulatory role in the invertebrate innate immune defense by regulating TLR and IL-16 expression.

The tuberous sclerosis complex 1 (TSC1) is a critical upstream regulator of autophagy. Nevertheless, the detailed molecular mechanism of TSC1 and its function in crustacean are still not well understood. In the present study, TSC1 was identified and characterized from Penaeus vannamei. Molecular feature analysis revealed that the open reading frame (ORF) of PvTSC1 was 3516 bp encoding a polypeptide of 1172 amino acids with a conserved N-terminal hamartin domain and two coiled-coil region domains in the C-terminus. The homologous alignment showed that the amino acid sequence of PvTSC1 shared 28.17% ~ 59.25% identity with that of TSC1 in other species. The mRNA transcripts of PvTSC1 were found in all detected tissues of shrimp, and PvTSC1 was mainly located in the cytoplasm of hemocytes. Antioxidant enzyme and PvTSC1 were significantly up-regulated in hemocytes, gill and hepatopancreas after high ammonia nitrogen stress. After PvTSC1 silencing, ROS accumulation increased and autophagy production decreased significantly. And the expression level of PvmTOR was increased significantly, on the contrary, autophagy-related genes (PvATG14 and PvBeclin-1) were decreased significantly. Finally, the mortality of shrimp increased significantly in the PvTSC1-silenced shrimp. These results indicated that ammonia nitrogen had a significant effect on oxidative stress injury of shrimp, and autophagy was a remarkable survival mechanism of shrimp with oxidative stress injury regulated by PvTSC1.

Males of the giant freshwater prawn (GFP), Macrobrachium rosenbergii, have been staged histologically into three morphotypes: small male (SM), orange claw (OC), and blue claw (BC). At present, the social population structure is the main limited factor of yield and profitability in commercial GFP production. The aim of this study is to investigate the relationship between the intestinal microbiota and the three male morphotypes of GFP. In this study, we quantified bacterial communities and abdominal muscle transcriptomes by parallel 16S rRNA gene amplicon sequencing and RNA-Seq in three male morphotypes of GFP. The abundance of bacteria varied significantly with three male morphotypes. Moreover, we identified 6062 differentially expressed genes and found a pattern of gene expression involved in developmental stages from the small male via the orange claw to the blue claw morphotype. Eleven signal transduction pathways classified into environmental information processing were related to the frequency of the different male morphotypes in the prawn population. Furthermore, the correlation analysis identified that p53, AMPK, MO25, and ATG1 gene expressions were positively correlated with the abundance of Candidatus Hepatoplasma and negatively correlated with the Lactococcus and Bacillus population in the giant freshwater prawn. We revealed an intimate linkage between the intestinal microbiome and morphotypic differentiation in the males of GFP. This study also provides a more effective method to manipulate the frequency of the different male morphotypes in the male prawn population.

Bax is a pro-apoptotic member of Bcl-2 like superfamily, playing an important role in regulating the apoptosis. In this study, the full-length Bax (EcBax) was obtained, containing a 5′UTR of 64 bp, an ORF of 579 bp and a 3′UTR of 1021 bp. The EcBax gene encoded a polypeptide of 192 amino acids with an estimated molecular mass of 21.55 KDa and a predicted isoelectric point (pI) of 6.75. The deduced amino acid sequence analysis showed that EcBax comprised the conserved residues and the characteristic domains known to the critical function of Bax. qRT-PCR analysis revealed that EcBax mRNA was broadly expressed in all of the examined tissues, while the highest expression level was observed in blood, followed by the expression in liver, gill, spleen, kidney, heart, muscle and intestine. A sharp increase of EcBax expression was observed in the vibrio challenge group by comparing with those in the control. Subcellular localization analysis revealed that EcBax was predominantly localized in the cytoplasm. EcBax exerted a regulatory role in modulating the mitochondrial membrane potential, promoting the cytochrome c release, and then activating the downstream caspase signaling. Moreover, the overexpression of EcBax can decrease the cell viability and antagonize NF-kB, AP-1, Stat3 promoter activity in Hela cells. These results indicate that EcBax containing the conserved domain of pro-apoptotic member of Bcl-2 family may disrupt the mammalian signaling and play a regulative role in the apoptotic process.

A novel turn-off fluorescent probe bearing a benzoxazole fluorophore has been synthesised and characterised. Its recognition properties towards different metal ions have been researched by spectrometry. The probe was highly selective and sensitive to Hg2+ and showed a complexation ratio towards Hg2+ of 1:1. Its fluorescence intensity varied almost linearly with the concentration of Hg2+ (1.6~8.6 μmol L−1), and the detection limit of Hg2+ was estimated to be 0.68 μmol L−1.

GRIM-19 (gene associated with retinoid-interferon-induced mortality 19), a novel cell death regulatory gene, plays important roles in cell apoptosis, mitochondrial respiratory chain and immune response. It has been reported to interact physically with STAT3 and inhibit STAT3-dependent signal transduction. In this study, a new GRIM-19 gene, which is a 789-bp gene encoding a 149 amino acids protein, is identified and characterized from Litopenaeus vannamei. The tissue distribution patterns showed that LvGRIM-19 was widely expressed in all examined tissues, with the highest expression in muscle. Quantitative real-time PCR revealed that LvGRIM-19 was down-regulated in hepatopancreas after infection with the Vibrio alginolyticus. Knockdown of LvGRIM-19 by RNA interference resulted in a lower mortality of L. vannamei under V. alginolyticus infection, as well as an enhancement in the protein expression of STAT gene and JAK gene. V. alginolyticus infection caused an increase apoptotic cell ratio and ROS production of L. vannamei, while LvGRIM-19 silenced shrimps showed significantly lower than GFP group. Our results suggest that the GRIM-19 plays a vital role in shrimps' responses to V. alginolyticus. Interferenced LvGRIM-19 treatment during V. alginolyticus infection could increase 12 h survival rate, which might indicated that LvGRIM-19 is closely related to death of shrimps.

Heavy metals are typical cumulative pollutants that can enter and poison the human body through the food chain. However, the molecular mechanism of heavy metal-induced oxidative stress is unclear. In this study, we characterize PvKelch-like-1 from P. vannamei and explore its antioxidant roles in immune regulation of crustaceans. PvKelch-like-1 full length contains 2107 nucleotides, consists of a 5&prime, untranslated region (UTR) of 79 bp, a 3&prime, UTR of 180 bp, and a ORF of 1848 encoded 615 amino acids, which contain a BTB, BACK and Kelch motif, putative molecular mass and isoelectric point were 69 KDa and 6.54. PvKelch-like-1 mRNA was ubiquitously expressed in all detected tissue of P. vannamei, and mRNA expression levels were significantly up-regulated from 6 to 24 h after cadmium stress and reached the highest level (3.2-fold) at 12 h in the hepatopancreas. Subcellular localization analysis revealed that PvKelch-like-1 was localized in the nucleus. Silencing PvKelch-like-1 significantly increased reactive oxygen species (ROS) (1.61-fold) production and DNA damage (1.32-fold) in the shrimp hemolymph, and significantly decreased total hemocyte counts (THC) (0.64-fold) at 6 h in hemolymph. Additionally, the antioxidant genes PvCAT (0.43-fold), PvMnSOD (0.72-fold), PvGST (0.31-fold) and PvGPx (0.59-fold) at 6 h were decreased significantly in PvKelch-like-1 silenced shrimp after cadmium stress. Overexpression of PvKelch-like-1 has the opposite results in enzyme activity. The SOD (2.44-fold) and CAT (2.19-fold) activities were significantly increased after overexpressing PvKelch-like-1. These results suggest that PvKelch-like-1 plays a vital role in shrimp innate immune defense by positively regulating the expression of antioxidant enzyme genes to respond to cadmium stress.

Scientific evidence is increasing that intestinal microbial flora is closely related to organism health and immunity. The relationship between the shrimp midgut microbial community and blue body syndrome is, however, unclear. In this study, we collected midgut and seawater samples from ponds with healthy and diseased shrimp to understand variations in bacterial communities in the midguts of healthy or diseased shrimp from the same habitat. The expression level of penaeidin, lectin, crustins, and defensin in shrimp with blue body syndrome were decreased, compared to those of healthy shrimp, indicating that shrimp with blue body syndrome have a lower autoimmune. 16 s sequencing analysis found that total 37 phyla, 91 classes, 160 orders, 221 families and 373 genera was detected. Among them, Proteobacteria (55.7%), Cyanobacteria (30.5%), Bacteroides (5.4%), Actinomycetes (1.1%) and Trichomes (0.9%) are the core bacteria in the intestinal bacteria of Penaeus vannamei. The intestinal bacterial communities are closely associated with the severity of shrimp disease. This pattern was further evidenced by NMDS and LEfSe analyses that were significantly different in abundance between healthy and diseased shrimp. In addition, we found that within a given bacterial genus, the patterns of enrichment or decrease were inconsistent with the known probiotics functions of those bacteria. These results suggested that the composition disorders and immunity decrease of intestinal microbiota were closely associated with shrimp blue body syndrome. Moreover, the identified bacterial genus can be used as an indicator taxa to evaluate the occurrence probability of blue body syndrome in shrimp.

CD59, a multifunctional glycoprotein, not only plays a regulatory role in complement cascades, but also participates in modulation of teleostean immunity. In this study, full length sequence of EcCD59 was obtained, comprising a 5'UTR of 163 bp, an ORF of 354 bp and a 3'UTR of 559 bp. EcCD59 gene encoded a polypeptide of 117 amino acids. Tissue-specific analysis revealed that the highest expression of EcCD59 mRNA was observed in muscle. Vibrio alginolyticus challenge can significantly increase EcCD59 mRNA expression in liver, kidney and spleen. EcCD59 distribution was detected by a combined approach using GFP-overexpression, immunofluorescence and ELISA assay, indicating that EcCD59 may be predominantly aggregated in cellular membrane. Both EcCD59 and EcCD59delGPI can directly bind to V. alginolyticus and decrease the in vitro growth of V. alginolyticus. Additionally, vibrio injection experiment indicated that the binding of EcCD59 or EcCD59delGPI to V. alginolyticus can restrict its growth rate in vivo. In this study, we found that EcCD59 may be involved in immune defense against vibrio infection in a complement-independent manner.

The critical behavior of the antiperovskite crystal Fe3.4In0.6N has been systematically studied in this paper. We have used a modified Arrott plot, the Kouvel-Fisher method, and critical isotherm analysis to obtain the critical exponents (β, γ and δ) which suggest that a long-range magnetic coupling described by mean-field (MF) theoretical model is dominant in Fe3.4In0.6N. The experimental M−T−H relations below and above Curie temperature (Tc) collapse into two branches by the single scaling equation m = f±(h) (m and h are renormalized magnetization and field), respectively. The critical exponents are confirmed by the Widom scaling law δ = 1 + γβ−1. In addition, it is suggested that the ferromagnetic (FM), anti-ferromagnetic (AFM) interactions and the magnetic disorders result to the exchange distance which is obtained just as J(r)∝ r - 4.4 .

Complement 1 inhibitor (C1INH) serving as a multifunctional factor can participate in the regulation of complement cascades and attenuate the activation of various proteases. In this study, we obtained EcC1INH cDNA and the tissue-specific analysis indicate that the highest expression level of EcC1INH mRNA was detected in liver. Moreover, Vibrio alginolyticus challenge can significantly increase EcC1INH mRNA expression in liver and kidney. N-terminal domain of EcC1INH could decrease LPS binding activity to cell surface, while loss of positively charged residues (PCRs) Arg21, His22, Lys50, Arg61 in N-terminal domain of EcC1INH can significantly reduce its interaction with LPS. Furthermore, LPS injection experiment indicated that the binding of EcC1INH N-terminal domain to LPS can antagonize LPS-induced inflammatory signaling pathway and attenuate the production of proinflammatory cytokines in vivo, indicating that EcC1INH was involved in negative regulation of inflammatory response.

It is well known that PI3K regulates various processes in mammalian cells by generating a secondary messenger that later activates AKT. However, its innate immune function in crustaceans remains unclear. We report the characterization of Litopenaeus vannamei PI3K (LvPI3K) for investigating how PI3K participates in the innate immunity of crustaceans. Full-length LvPI3K cDNA was 3357 bp long, with a 3222 bp open reading frame (ORF) that encodes a putative protein of 1292 amino acids. The PI3K catalytic domain (PI3Kc) of LvPI3K was found to be rather conserved when the PI3Ks from other species were analyzed. The LvPI3K protein was shown to be localized to the cytoplasm of Drosophila S2 cells, while LvPI3K mRNA was ubiquitously expressed in healthy L. vannamei, with the highest expression found in hemolymph. A dual luciferase reporter gene assay demonstrated that LvPI3K overexpression activated the promoter of antibacterial peptide LvPEN4 in a dose-dependent manner. However, the addition of PDTC, a specific inhibitor of NF-κB, suppressed the LvPI3K-induced LvPEN4 promoter activation. Moreover, Vibrio alginolyticus challenge induced a rapid up-regulation of LvPI3K expression. Further experiments showed that LvPI3K silencing in shrimp challenged with V. alginolyticus significantly increased Vibrio number, ROS production and DNA damage in the hemolymph, as well as significantly decreased total hemocyte count. The mRNA levels of certain molecules related to LvPI3K signaling, such as LvAKT and LvPEN4, also decreased following LvPI3K silencing. Taken together, these results suggest that LvPI3K regulates the downstream signal component LvPEN4 and functions in V. alginolyticus resistance.

Vibrio alginolyticus containing the highly toxic extracellular product is one of the most serious threats to grouper survival and its minimum lethal dose is approximately 500 CFU/g fish body weight in grouper. To study the toxic effects of V. alginolyticus on the immune system in teleost, Calmodulin (CaM), an important molecular indicator gene, was cloned from the orange-spotted grouper (Epinephelus coioides). The full-length Ec-CaM consisted of a 5'-UTR of 103 bp, an ORF of 450 bp and a 3'-UTR of 104 bp. The Ec-CaM gene encoded a protein of 149 amino acids with an estimated molecular mass of 16.4 kDa and a predicted isoelectric point of 3.93. The deduced amino acid sequence showed that Ec-CaM contained four highly conserved EF-hand domains known to be critical for the function of CaM. Ec-CaM was widely expressed and the highest expression level was observed in liver. Following V. alginolyticus challenge, a sharp increase level of respiratory burst activity and apoptosis ratio were observed. Further analyses of CaM expression and p53 expression in liver, kidney and spleen by qRT-PCR demonstrated that the up-regulated expression of CaM and p53 were observed in the vibrio challenge group. Western blotting analysis confirmed that the Ec-CaM protein was strongly induced in liver at 12 h post-injection, while a sharp increase of p53 protein expression was observed at 24 h post-injection. These results showed CaM expression serving as a potential molecular indicator may help to assess the toxicological effects of V. alginolyticus on the ROS generation and apoptotic process in grouper.

Heavy metal residues and chemical contaminators considered as relevant sources of aquatic environmental pollutants have a generally immunosuppressive effect on aquatic organisms, depressing metabolic activities and immune response. Glutamine: fructose-6-phosphate aminotransferase (GFAT, EC2.6.1.16) is the first, and rate-limiting, enzyme in the hexosamine biosynthetic pathway, and is involved in the regulation of chitin biosynthesis and glycosylation of proteins. We have isolated and characterized GFAT from the white shrimp Litopenaeus vannamei. Amino acid sequence similarity of the Lv-GFAT (L.vannamei-GFAT) was highest to GFATs isolated from insects and mammals (83 % similarity to that of Haemaphysalis longicornis). The open-reading frame of the Lv-GFAT codes for a protein of 41.6 kDa with a calculated isoelectric point of 5.03. RT-PCR assays showed that endogenous Lv-GFAT mRNA is most strongly expressed in the intestine. Further analysis of Lv-GFAT gene expression in hepatopancreas by quantitative real-time PCR demonstrated that Lv-GFAT transcript levels increased when the shrimp were exposed to alkaline pH (9.3) and cadmium stress, but the time when its mRNA expression level peaked differed under these stresses. We also first expressed the recombinant protein of GFAT from shrimps in Escherichia coli. Western blot analyses confirmed that the Lv-GFAT protein was strongly expressed in the hepatopancreas after exposure to the LC-Cd stress. These results suggest that Lv-GFAT expression is stimulated by alkaline pH and cadmium stress and that it may play important roles in resistance of shrimp to environmental stresses.