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In recent decades, global aquaculture has expanded rapidly, raising concerns about coastal environmental degradation due to unregulated or poorly regulated discharge of aquaculture tailwater. Despite the crucial role of dissolved organic matter (DOM) in biogeochemical processes and aquatic biodiversity, the influence of aquaculture type on the molecular characteristics of DOM remains largely unexplored. Herein, this study investigated the variations in chemical and spectroscopic properties as well as molecular characteristics and composition of DOM across different aquaculture types including crustacean, fish and shellfish. Our findings revealed notable differences in DOM quantities among different aquaculture types, with crustacean and fish aquaculture water containing higher DOM amount compared to shellfish aquaculture water. This disparity can be attributed to the more frequent formulated feeds of crustacean and fish in contrast to shellfish aquaculture. Furthermore, distinct differences were also observed in the characteristics and composition of DOM among the different aquaculture waters. Specifically, DOM in shellfish aquaculture water exhibited a higher abundance of unsaturated and reduced molecules as well as increased aromaticity compared to the other two aquaculture waters. Conversely, DOM from fish aquaculture water showed a greater contribution from terrestrial origin characterized by elevated levels of plant-based components such as lignin-like and tannin-like compounds. Interestingly, DOM from shellfish aquaculture water contained lower levels of microbial-derived components such as lipid-like and protein-like compounds, likely due to reduced microorganism populations resulting from lower nutrients availability and higher salinity. Overall, these significant variations in characteristics and composition of DOM underscore the potential impacts of aquaculture type on the DOM biogeochemical cycle and the environmental quality in aquatic ecosystems., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Trace organic contaminants （TrOCs） in reclaimed water can pose potential ecological and health risks during long-term reuse, but the current water quality standards have not restricted their limits. For preventing and controlling the potential risks from TrOCs, an integrated method EHL for identifying priority pollutants in reclaimed water was proposed. This method followed three steps： First, a meta-analysis was performed to assess the exposure of TrOCs in reclaimed water, and a database of TrOCs exposure （E） was established. Then, those hazardous pollutants of particular concern were selected from the E database based on the consensus mechanism, and a dataset of hazardous TrOCs （H） in reclaimed water was established. Finally, the risk quotient of candidates in the H dataset was calculated, and according to the risk-based prioritizing, a final list of priority TrOCs （L） in reclaimed water was recommended. To determine the procedure of EHL, the priority pollutants for reclaimed water in China were identified. Based on literature data from the past 30 years, a total of 32 TrOCs of high concern were selected. Six TrOCs with the highest risk were recommended as an executive priority list for reclaimed water management, including formaldehyde, bis（2-ethylhexyl） phthalate （DEHP）, 17 β -estradiol, erythromycin, sulfamethoxazole, and ibuprofen. The EHL method could effectively identify the priority pollutants in reclaimed water and other water environments, and it is expected to play a crucial role in water quality safety evaluation and management.

The goal of environmental management in China is transitioning from pollution control to the improvement of ecological quality. The establishment of regional differential water quality standards can better adapt to the ecological characteristics and development needs of different regions. Given the shortcomings of current water quality standards and the needs of technological development, this study analyzed the causes and influencing factors of regional differences in water quality standards and summarized China's regional variations in areas such as the characteristics of receiving water bodies, social attributes, climate conditions, physicochemical properties of water, and aquatic biotic populations. It also examined the impact of regional characteristics on the assessment of biological toxicity. Based on these analyses, the study identified key scientific questions that may arise in the development of regional differential water quality standards in China and outlined future critical research directions in this field. These directions include developing theories and methods for determining the effluent dilution coefficient, establishing methods for identifying regional key water quality indicators and assessing their impact on water quality standards, determining regional characteristic species and their influence on water quality standards, and formulating theories and methods for categorizing different regions across the country. The aim of the study was to provide a reference for the scientific establishment of water quality standards in China.

The giant jellyfish Nemopilema nomurai sting can cause local and systemic reactions; however, comparative analysis of the tentacle extract (TE) and nematocyst venom extract (NV), and its toxicity, mechanism, and potential intervention are still limited. This study compared venom from TE and NV for their composition, toxicity, and efficacy in vitro and in vivo used RAW264.7 cells and ICR mice. A total of 239 and 225 toxin proteins were identified in TE and NV by proteomics, respectively. Pathological analysis revealed that TE and NV caused heart and liver damage through apoptosis, necrosis, and inflammation, while TE exhibited higher toxicity ex vivo and in vivo. Biochemical markers indicated TE and NV elevated creatine kinase, lactatedehydrogenase, and aspartate aminotransferase, with the TE group showing a more significant increase. Transcriptomics and Western blotting indicated both venoms increased cytokines expression and MAPK signaling pathways. Additionally, 1 mg/kg PACOCF3 (the phospholipase A2 inhibitor) improved survival from 16.7% to 75% in mice. Our results indicate that different extraction methods impact venom activities, tentacle autolysis preserves toxin proteins and their toxicity, and PACOCF3 is a potential antidote, which establishes a good extraction method of jellyfish venom, expands our understanding of jellyfish toxicity, mechanism, and provides a promising intervention.

This study primarily focused on the regional disparities in both water quality criteria and ecological risks attributed to cadmium presence within the surface waters of the Yangtze River Basin. In the initial phase, the long-term water quality criteria for cadmium were recalibrated in accordance with the guidelines outlined in China's "Water Quality Criteria for Freshwater Aquatic Organisms-Cadmium," accounting for the prevalent hardness distribution within the Yangtze River Basin's surface water. Subsequently, a more refined revision was undertaken considering the specific characteristics of the species residing within the Yangtze River Basin. This undertaking led to a comprehensive interpretation of the regional variations in both the distribution of long-term water quality criteria values and the risk quotient distribution of cadmium throughout the Yangtze River Basin. The incorporation of hardness and species-specific attributes resulted in a revised range of long-term water quality criteria for cadmium across different urban locales within the Yangtze River Basin. Notably, the recalibrated values ranged from 0.08 μg·L -1 as the lowest threshold to 0.75 μg·L -1 as the upper limit, signifying a tenfold differentiation. Correspondingly, the urban average annual risk quotient associated with cadmium exposure demonstrated a variation from 0.035 to 1.12, marking a significant 32-fold discrepancy between the lowest and highest values. It is essential to highlight that regions of paramount importance, such as the confluence area connecting the upper and middle stretches of the Yangtze River Basin and the intricate Dongting Lake system, exhibited noteworthy ecological risks attributed to cadmium presence. Consequently, further in-depth investigations into these critical regions are imperative for a comprehensive understanding of the associated risks.

Changes in protein abundance and reversible protein phosphorylation (RPP) play important roles in regulating hypometabolism but have never been documented in overwintering frogs at high altitudes. To test the hypothesis that protein abundance and phosphorylation change in response to winter hibernation, we conducted a comprehensive and quantitative proteomic and phosphoproteomic analysis of the liver of the Xizang plateau frog, Nanorana parkeri , living on the Qinghai-Xizang (Tibet) Plateau (QTP). In total, 5 170 proteins and 5 695 phosphorylation sites in 1 938 proteins were quantified. Based on proteomic analysis, 674 differentially expressed proteins (438 up-regulated, 236 down-regulated) were screened in hibernating N. parkeri versus summer individuals. Functional enrichment analysis revealed that higher expressed proteins in winter were significantly enriched in immune-related signaling pathways, whereas lower expressed proteins were mainly involved in metabolic processes. A total of 4 251 modified sites (4 147 up-regulated, 104 down-regulated) belonging to 1 638 phosphoproteins (1 555 up-regulated, 83 down-regulated) were significantly changed in the liver. During hibernation, RPP regulated a diverse array of proteins involved in multiple functions, including metabolic enzymatic activity, ion transport, protein turnover, signal transduction, and alternative splicing. These changes contribute to enhancing protection, suppressing energy-consuming processes, and inducing metabolic depression. Moreover, the activities of phosphofructokinase, glutamate dehydrogenase, and ATPase were all significantly lower in winter compared to summer. In conclusion, our results support the hypothesis and demonstrate the importance of RPP as a regulatory mechanism when animals transition into a hypometabolic state.

The plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzoniae) are native species unique to the Qinghai-Tibetan Plateau with successful adaptation to the hypoxic environment. In this study, the number of red blood cells, hemoglobin concentration, mean hematocrit and mean volume of red blood cells were measured in plateau zokors and plateau pikas at different altitudes. Hemoglobin subtypes of two plateau animals were identified by mass spectrometry sequencing. The forward selection sites in two animals' hemoglobin subunits were analyzed by PAML4.8 program. Homologous modeling was used to analyze the effect of forward selection sites on the affinity of hemoglobin to oxygen. The adapting strategies of plateau zokors and plateau pikas to hypoxia at different altitudes were analyzed through comparing blood parameters between the two species. The results indicated that, with increasing altitudes, plateau zokors responded to hypoxia by increasing red blood cell count and decreasing red blood cell volume, while plateau pikas took the opposite strategies to plateau zokors. In erythrocytes of plateau pikas, both adult α 2 β 2 and fetal α 2 ε 2 hemoglobins were identified, while erythrocytes of plateau zokors only had adult α 2 β 2 hemoglobin, however the affinities and the allosteric effects of the hemoglobin of plateau zokors were significantly higher than those of plateau pikas. Mechanistically, in the α and β subunits of hemoglobin of plateau zokors and pikas, the numbers and the sites of the positively selected amino acids as well as the side chain groups polarities and orientations of the amino acids differed significantly, which may result in the difference of the affinities to oxygen of hemoglobin between plateau zokors and pikas. In conclusion, the adaptive mechanisms to respond to hypoxia in blood properties of plateau zokors and plateau pikas are species-specific.

Wastewater reclamation and reuse are an effective measure to alleviate water shortages. Water quality stability is the premise for safe utilization of reclaimed water and other water resources. Stable water quality can prevent the frequent occurrence of corrosion and scaling, which not only improves the sensory properties of water quality but also reduces secondary pollution and energy consumption. In order to promote the utilization of wastewater resources, it is important to evaluate and manage reclaimed water quality stability. Based on substantial literature and research, the characteristics and scopes of evaluation parameters for both chemical stability and biological stability were summarized, and a set of evaluation parameters and a system for reclaimed water quality stability were comprehensively established in this study. The evaluation procedure was mainly divided into four steps including:① determine the influencing factors of reclaimed water quality stability; ② select corresponding evaluation parameters based on reclaimed water quality characteristics, the materials of pipelines, and equipment; ③ evaluate the reclaimed water quality stability; and ④ examine whether the evaluation results were appropriate. Finally, the potential challenges for the stability evaluation of water quality were analyzed. This study is expected to provide a theoretical foundation for the scientific evaluation of water quality stability and safe utilization of reclaimed water.

Emerging trace organic contaminants (TOrCs) have been ubiquitously detected in municipal wastewater. TOrCs show low concentrations but are usually harmful to aquatic biota and recalcitrant to treatments. The advanced treatment of TOrCs is challenged by four main aspects. First, it is hard to qualify the chemical structure and concentrations of TOrCs, which are typically very low. Second, risk assessment results vary with the selection of target-species and toxicity-endpoints, which makes it hard to regulate priority lists and concentration limits. Third, TOrCs account for less than 0.01% of the total organics in wastewater. Thus, TOrCs cannot be selectively and efficiently removed by conventional treatments. Fourth, it is hard to optimize and manage the treatment trains of TOrCs. In this manuscript, a new strategy combing non-targeting TOrC screening and TOrC "fingerprinting" are proposed to overcome the difficulties of quantification and treatment optimization. Based on this, the standards and regulations to control the TOrCs in reclaimed water should be developed on the basis of elimination efficiency rather than concentration.

In the present study, the composition and content of pulmonary surfactant (PS) were analyzed to explore the hypoxia adaptation mechanism in plateau zokors (Myospalax baileyi) and plateau pikas (Ochotona curzoniae). 36 plateau zokors and plateau pikas were trapped alive at the Laji Mountain in Guide County, Qinghai Province (at the altitude of about 3 600 m), and 36 Sprague-Dawley (SD) rats were purchased from the experimental animal center of Lanzhou University (at the altitude of about 1 500 m). All animals were lavaged after laboratory anesthesia, the blood in lung tissues was fully washed out and the lung tissues were then taken out to obtain the bronchoalveolar lavage fluid by bronchoalveolar lavage. The composition and content of phospholipids in the PS of three different kinds of animals were analyzed by using high performance liquid chromatography; the protein composition, content and types in the PS were analyzed by G-250 Coomassie brilliant blue method, polyacrylamide gel electrophoresis (PAGE) and mass spectrometry; the dissolved oxygen in the PS solutions were determined by using dissolved oxygen electrode. The results showed that the total contents of phospholipids in the PS were successively increased among plateau zokors, plateau pikas and SD rats (P < 0.05), while the total content of proteins successively decreased (P < 0.05). There were five phospholipids identified in the PS, including linoleic palmitoylphosphatidylcholine (LPPC), dipalmitoylphosphatidylcholine (DPPC), phosphatidylglyerol (PG), phosphatidylinositol (PI) and phosphatidylserine (PSe), but the relative contents of these phospholipids were different. The relative content of LPPC was successively increased among plateau zokors, plateau pikas and SD rats (P < 0.01). The relative contents of DPPC, PG and PI in the PS of plateau zokors were significantly higher than those of plateau pikas (P < 0.01), while insignificant differences between plateau pikas and SD rats (P > 0.05). The relative content of PSe had no significant differences between plateau zokors and plateau pikas (P > 0.05), but both were significantly higher than that of SD rats (P < 0.01). The serum albumin (SA) was identified in the PS of three kinds of animals, including homologous tetramer protein containing heme, which is composed of hemoglobin β subunit, in plateau zokors and plateau pikas. Immunoglobulin (Ig) heavy chain was found in PS of plateau zokors and SD rats. The content of Ig heavy chain in plateau zokor was significantly higher than that in SD rats (P < 0.01), and the content of protein containing heme was significantly higher than that in plateau pikas (P < 0.05). The amount of dissolved oxygen was successively decreased in the PS among plateau zokors, plateau pikas and SD rats (P < 0.01), but it was significantly higher than that in saline (P < 0.01). These results suggest that the total content of proteins in the PS of plateau zokors and plateau pikas was significantly higher, while the total content of phospholipids was significantly decreased. There were high content of homologous tetramer protein containing heme in the PS of plateau zokors and plateau pikas. The relative content of DPPC, the main component of phospholipids, was significantly increased in plateau zokors. The changes of PS component and content improve the adaptability of the two plateau animals in hypoxia environment.

The assay was used in HIV/AIDS surveillance sentinel sites to identify recent HIV-1 infection, to estimate HIV-1 incidence and understand the epidemic trends among men who have sex with men during 2011-2015. During 2011-2015, the HIV-1 incidence of men who have sex with men was 5.16%(95 %CI : 4.65%-5.66%) in Sichuan Province. According to, a total of 9 cities had been divided into two type of regions: the first type(the proportion of live HIV/AIDS accounts for more than 0.1% of the permanent population, and the number of live HIV/AIDS has reached 500 cases and above) and the second type(the proportion of live HIV/AIDS accounts for 0.03% to 0.100% of the permanent population, and the number of live HIV/AIDS has reached 100-500 cases). During 2011-2015, the HIV-1 incidence of the second type was 4.24% (95 %CI : 3.72%-4.76%). The HIV-1 incidence of men who have sex with men was at a high level. The spread of HIV-1 to the second region was possible.

Objective: Data from the HIV/AIDS surveillance sentinel sites was used to identify the recent HIV-1 infections and to estimate the HIV-1 incidence and epidemic trends among intravenous drug users (IDU), MSM, female sex workers and men attending the sexually transmitted disease clinics in Sichuan province. Methods: Specimens collected from IDUs, MSM, female sex workers and men attending the sexually transmitted disease clinics sentinel sites were confirmed as HIV-1 positive and tested with BED-CEIA. Annual HIV-1 prevalence and incidence rates were calculated and the incidence was adjusted by McNermar method. Results: A total number of 194 223 serum specimens were collected and HIV antibody tested, with 5 297 HIV-1 positive. 4 640 of the positives underwent BED-CEIA, with 749 identified as recent infections. During 2011-2015, HIV-1 incidence rates of the four types of population appeared as 5.16% (95% CI : 4.65-5.66), 0.22% (95% CI :0.16-0.28), 0.57%(95% CI : 0.45-0.69), 7.53% (95% CI : 6.06-9.01) and 0.44% (95% CI : 0.36-0.53) respectively. Significant differences were seen in the HIV-1 incidence rates. Conclusions: The HIV-1 incidence rates of IDUs from other sentinel sites in Sichuan province, female sex workers and men attending the sexually transmitted disease clinics appeared low, showing that the AIDS prevention and control activities worked effectively. However, the HIV-1 incidence of MSM was still at high level and even rising, suggesting that HIV-1 transmission between MSM and IDUs was possible.

The plateau pika ( Ochotona curzoniae ) is a species of sprint-running alpine animals in the Qinghai-Tibet Plateau, which is a harsh highland hypoxic environment. Ldh-c is expressed in the testis, sperm and somatic tissues of plateau pika. To reveal the role and physiological mechanisms of sperm-specific lactate dehydrogenase (LDH-C 4 ), in plateau pika to adapt to hypoxic environment, an adenoviral line of pMultiRNAi- Ldhc was constructed and injected into the bilateral biceps femoris of the hind legs. The swimming times of the pikas, and the Ldh-c expression levels, total LDH activities and ATP levels in skeletal muscle, were measured after the pikas were raised in the trapped site for 5 days. Our results showed that after Ldh-c was silenced, the sprint-running ability (swimming time) of the plateau pikas was significant decreased, and the total LDH activities and ATP levels were reduced by 28.21% and 27.88%, respectively. Our results indicated that expression of Ldh-c in the skeletal muscle of plateau pika increased anaerobic glycolysis and enhanced adaptation to highland hypoxic environments., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2017. Published by The Company of Biologists Ltd.)

In this study, we optimized a restriction-ligation-free (RLF) method to save time and cost of constructing multiple plasmids with the same gene insert, and examined the efficacy of RLF on high-throughput multi-plasmid cloning. This method utilizes the precise DNA repair and recombination systems within Escherichia coli, which allows to bypass the in vitro restriction and ligation enzyme reactions commonly included in routine cloning procedures. A homologous arm is linked to the 5'-end of the forward primer used to amplify both the target gene and vector. A different homologous arm is linked to the 5'-end of the reverse primer. Therefore, genes can be cloned into the vectors by homologous recombination after co-transformation of the amplified target gene and the linearized vector, which bear the same homologous arm on either end. More than twenty-four different plasmids were generated by this method, which uses two simple polymerase chain reaction steps. This method is highly efficient in cloning any gene of interest into any vector at any site without sequence constraints, as no restriction and ligation reactions are required.

Objective: To explore the intergrating of N-isopropyl oxamate and serum protein and establish a high performance liquid chromatography (HPLC) detection method of N-isopropyl oxamate (specific inhibitor of testis-specific lactate dehydrogenase (LDH-C4)) in the blood of plateau pikas., Methods: Twenty highland pika 150-200 g, were randomly divided into two groups (n = 10): control group and inhibitor group. Different concentrations of N-isopropyl oxamate were added to examine the intergrating of N-isopropyl oxamate and serum protein. In order to determine its concentration in the pika blood accurately, we used the method of adding trypsin to incubate the serum first, followed by trichloroacetic acid treatment and detecting by HPLC. Results: When the concentrations of N-isopropyl oxamate in the pika serum were added to 0.05 mmol/L, 0.1 mmol/L, 1 mmol/L, 10 mmol/L, 16.7 mmol/L, 33.3 mmol/L and 100 mmol/L, the intergrating rates between N-isopropyl oxamate and plateau pika serum were 100%, 100%, 100%, 86.84%, 54.11%, 40.10% and 20.18%, respectively. The method established in this paper was good on recovery rates, precision and stability. A good linearity was obtained in the range of 0.0125-0.25 mmol/L. When the concentrations of N-isopropyl oxamate in the serum were added to 0. 15 mmol/L,0.3 mmol/L and 1 mmol/L, the recovery rates were 98.05%, 98.98% and 98.12%, respectively; the precision relative standard deviation( RSD) of concentrations were 1.17%, 0.92% and 0.83%, respectively; the stability relative standard deviation (RSD) of concentrations were 1.38%, 1.40% and 0.88%, respectively. The repeatability RSD of the method was 1.76%. Quantitative limit was 0.0125 mmol/L., Conclusion: N-isopropyl oxamate has a strong affinity with plateau pika serum protein that can't be accurately determined with common HIPLC method. It can be accurately determined in the blood by adding trypsinto digest the serum protein first, followed by adding trichloroacetic acid to precipitate the protein.

The plateau pika (Ochotona curzoniae) has a strong adaptability to hypoxic plateau environment. We found that the sperm-specific lactate dehydrogenase (LDH-C4) gene Ldh-c expressed in plateau pika cardiac muscle. In order to shed light on the effect of LDH-C4 on the anaerobic glycolysis in plateau pika cardiac muscle, 20 pikas were randomly divided into the inhibitor group and the control group, and the sample size of each group was 10. The pikas of inhibitor group were injected with 1 mL 1 mol/L N-isopropyl oxamate, a specific LDH-C4 inhibitor, in biceps femoris muscle of hind legs, each leg with 500 μL. The pikas of control group were injected with the same volume of normal saline (0.9% NaCl). The mRNA and protein expression levels of Ldh-c gene in plateau pika cardiac muscle were determined by real-time PCR and Western blot. The activities of LDH, and the contents of lactate (LD) and ATP in cardiac muscle were compared between the inhibitor group and the control group. The results showed that 1) the expression levels of Ldh-c mRNA and protein were 0.47 ± 0.06 and 0.68 ± 0.08, respectively; 2) 30 min after injection of 1 mL 1 mol/L N-isopropyl oxamate in biceps femoris muscle, the concentration of N-isopropyl oxamate in blood was 0.08 mmol/L; 3) in cardiac muscle of the inhibitor group and the control group, the LDH activities were (6.18 ± 0.48) U/mg and (9.08 ± 0.58) U/mg, the contents of LD were (0.21 ± 0.03) mmol/g and (0.26 ± 0.04) mmol/g, and the contents of ATP were (4.40 ± 0.69) nmol/mg and (6.18 ± 0.73) nmol/mg (P < 0.01); 5) the inhibition rates of N-isopropyl oxamate to LDH, LD and ATP were 31.98%, 20.90% and 28.70%, respectively. The results suggest that Ldh-c expresses in cardiac muscle of plateau pika, and the pika cardiac muscle may get at least 28% ATP for its activities by LDH-C4 catalyzed anaerobic glycolysis, which reduces the dependence on oxygen and enhances the adaptation to the hypoxic environments.