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Abstract This study was conducted to investigate the energy mobilisation preference and ionoregulation pattern of female tilapia, Oreochromis sp. living in different environments. Three different treatments of tilapia as physiology compromising model were compared; tilapia cultured in recirculating aquaculture system (RAS as Treatment I—RAS), tilapia cultured in open water cage (Treatment II—Cage) and tilapia transferred from cage and cultured in RAS (Treatment III—Compensation). Results revealed that tilapia from Treatment I and III mobilised lipid to support gonadogenesis, whilst Treatment II tilapia mobilised glycogen as primary energy for daily exercise activity and reserved protein for growth. The gills and kidney Na+/K+ ATPase (NKA) activities remained relatively stable to maintain homeostasis with a stable Na+ and K+ levels. As a remark, this study revealed that tilapia strategized their energy mobilisation preference in accessing glycogen as an easy energy to support exercise metabolism and protein somatogenesis in cage culture condition, while tilapia cultured in RAS mobilised lipid for gonadagenesis purposes.

The ongoing climate change is resulting in elevating temperatures and reduced pH levels in the marine environment. This work reports on the interactive effects of these two factors on growth and physiological responses of Pacific whiteleg shrimps (Litopenaeus vannamei), with a very important commercial shrimp specie. The study examines the interactive effects of temperature (28 versus34°C) and pH (6 versus 8) were examined at four combinations: (28°C and pH 8 (N28); 34°C and pH 8 (N34); 28°C and pH 6 (A28); and 34°C and pH 6 (A34), over a 30 days period. A combination of a high temperature and low pH (i.e., A34) was found to impose a high metabolic burden on the shrimp, resulting in an increased food intake and elevated osmorespiration responses relative to control, but no improvement in growth. The A34 stress combination increased ammonia excretion 2-fold relative to control, and increased a 40 % rise in metabolic oxygen uptake. Under A34 stress, glycogen and lipid reserves in both muscle and hepatopancreas were mobilized to support basal metabolic needs, resulting in growth suppression. The combined thermal-pH stress A34 did not affect the calcium and magnesium content of muscle and shell, which are essential minerals for exoskeleton and structural development. Although the shrimp can survive under high temperature-low pH stress conditions, their growth rate was compromised.

High alkaline environment can lead to respiratory alkalosis and ammonia toxification to freshwater fish. However, the Amur ide (Leuciscus waleckii), which inhabits an extremely alkaline lake in China with titratable alkalinity up to 53.57 mM (pH 9.6) has developed special physiological and molecular mechanisms to adapt to such an environment. Nevertheless, how the Amur ide can maintain acid-base balance and perform ammonia detoxification effectively remains unclear. Therefore, this study was designed to study the ammonia excretion rate (Tamm), total nitrogen accumulation in blood and tissues, including identification, expression, and localization of ammonia-related transporters in gills of both the alkali and freshwater forms of the Amur ide. The results showed that the freshwater form Amur ide does not have a perfect ammonia excretion mechanism exposed to high-alkaline condition. Nevertheless, the alkali form of Amur ide was able to excrete ammonia better than freshwater from Amur ide, which was facilitated by the ionocytes transporters (Rhbg, Rhcg1, Na+/H+ exchanger 2 (NHE2), and V-type H+ ATPase (VHA)) in the gills. Converting ammonia into urea served as an ammonia detoxication strategy to reduced endogenous ammonia accumulation under high-alkaline environment.

Abstract Background Porcine epidemic diarrhea virus (PEDV) and porcine delta-coronavirus (PDCoV) are economically important pathogens that cause diarrhea in sows and acute death of newborn piglets. Moreover, the emerging PDCoV was reported to infect children. The current situation is that vaccine prevention has not met expectations, and emergency containment strategies following outbreaks cannot prevent the damages and losses already incurred. Therefore, a more sensitive detection method, that is both convenient and enables accurate and effective sequencing, that will provide early warning of PEDV and PDCoV is necessary. This will enable active, effective, and comprehensive prevention and control, which will possibly reduce disease occurrences. Results Duplex nested RT-PCR (dnRT-PCR) is an ideal method to achieve early warning and monitoring of PEDV and PDCoV diseases, and to additionally investigate any molecular epidemiological characteristics. In this study, two pairs of primers were designed for each virus based upon the highly conserved N protein sequences of both PEDV and PDCoV strains retrieved from the NCBI Genbank. After optimization of the reaction conditions, the dnRT-PCR assay amplified a 749-bp fragment specific to PEDV and a 344-bp fragment specific to PDCoV. Meanwhile, the specificity and sensitivity of the primers and clinical samples were tested to verify and establish this dnRT-PCR method. The limit of detection (LoD)for both PEDV and PDCoV was 10 copies/µL. The results showed that among 251 samples, 1 sample contained PEDV infection, 19 samples contained a PDCoV infection, and 8 samples were infected with both viruses, following the use of dnRT-PCR. Subsequently, the positive samples were sent for sequencing, and the sequencing results confirmed that they were all positive for the viruses detected using dnRT-PCR, and conventional RT-PCR detection was conducted again after the onset of disease. As these results were consistent with previous results, a detection method for PEDV and PDCoV using dnRT-PCR was successfully established. In conclusion, the dnRT-PCR method established in this study was able to detect both PEDV and PDCoV, concomitantly. Conclusions The duplex nested RT-PCR method represents a convenient, reliable, specific, sensitive and anti-interference technique for detecting PEDV and PDCoV, and can additionally be used to simultaneously determine the molecular epidemiological background.

Current water warming and freshwater acidification undoubtedly affect the life of aquatic animals especially ammonotelic teleost by altering their physiological responses. The effect of temperature (28 °C vs 32 °C) and pH (7 vs. 5) on the metabolic compromising strategies of Hoven's carp (Leptobarbus hoevenii) was investigated in this study. Fishes were conditioned to (i) 28 °C + pH 7 (N28°C); (ii) 32 °C + pH 7 (N32°C); (iii) 28 °C + pH 5 (L28°C) and (iv) 32 °C + pH 5 (L32°C) for 20 days followed by osmorespiration assay. Results showed that feeding performance of Hoven's carp was significantly depressed when exposed to low pH conditions (L28°C and L32°C). However, by exposed Hoven's carp to L32°C induced high metabolic oxygen intake and ammonia excretion to about 2x-folds higher compared to the control group. As for energy mobilization, Hoven's carp mobilized liver and muscle protein under L28°C condition. Whereas under high temperature in both pH, Hoven's carp had the tendency to reserve energy in both of liver and muscle. The findings of this study revealed that Hoven's carp is sensitive to lower water pH and high temperature, thereby they remodeled their physiological needs to cope with the environmental changes condition.

With the development of computer-aided surgery and rapid prototyping via 3D printing technology, digital surgery has rapidly advanced in clinical practice, especially in the field of oral and maxillofacial surgery. 3D printing technology has been applied to the functional restoration and reconstruction of the jawbone. Before surgery, a 3D digital model is constructed through software to plan the scope of the osteotomy, shape the bone graft and plan the placement of the implant. Additionally, 3D models of personalized surgical instrument guides are printed prior to surgery. With these 3D-printed models and guides, accurate excision of the jaw tumor, accurate placement of the grafted bone and precise placement of implants can be achieved during surgery. Postoperative evaluation of accuracy and function shows that 3D printing technology can aid in achieving the biomechanical goals of simultaneous implant placement in jaw reconstruction, and in combination with dental implant restoration, the technology can improve patients' postoperative occlusal and masticatory functions. Nevertheless, 3D printing technology still has limitations, such as time-consuming preparation before surgery. In the future, further development of 3D printing technology, optimization of surgical plans, and alternative biological materials are needed. Based on domestic and foreign literature and our research results, we have reviewed the process and clinical application prospects of jaw reconstruction via 3D printing technology to provide a reference for oral and maxillofacial surgeons.

Modular multilevel converter (MMC), which is composed of a large number of submodules (SM) connected in series, is one of the most promising converters in high voltage field. The open-circuit fault and short-circuit fault of the SM's power switch seriously affect the reliability of the MMC, and it brings great challenges to the stable operation of the MMC. Therefore, in order to quickly detect the faulty SM, a fault detection method of the SM's power switch in the MMC based on Luenberger observer is proposed. Firstly, the fault characteristic of the SM is analyzed, and the model of Luenberger observer is established according to the mathematical relation of the SM capacitor voltage change. Then, the estimated capacitor voltage is calculated by Luenberger observer, and the detection of the power switch fault is realized by comparing the estimated capacitor voltage and the measured capacitor voltage. Finally, the simulation model of the MMC system based on the PSCAD/EMTDC and the experiment platform in the laboratory are respectively built. The simulation and experiment results show that the open-circuit and short-circuit fault of the SM can be detected correctly and effectively, which verifies the validity and feasibility of the proposed method, but each SM needs to be monitored.

Unified power flow controller (UPFC), static synchronous series compensator (SSSC), thyristor controlled series compensation (TCSC) and phase shifter (PST) can be able to adjust line parameters and flexibly control power flow. They have been applied in domestic and foreign power grids, and have played an important role in balancing power flow in transmission channels and improving power supply capacity. The theoretical research and engineering practice of UPFC, SSSC, TCSC and PST are summarized. Firstly, the basic structure and power flow control principle of four kinds of power flow control devices are analyzed, and projections of power flow control devices are introduced. Then, the key technologies of power flow control devices are summarized from five aspects: topology, converter technology, location and capacity, control strategy and fault protection. Finally, the typical application scenarios of power flow control devices in the future power grid are prospected, and the research direction of power flow control technology in the future is analyzed.

Objective To investigate the antibacterial properties, biocompatibility and mechanical properties of Cu-ZnO-loaded dental veneering porcelain to provide an experimental basis for the development of new dental veneering porcelain. Methods Cu-ZnO nanoparticles were added to IPS E.max Ceram for restorative veneer porcelain at different mass percentages of 0 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, and 6 wt% using ball milling in ceramic powder. A cylindrical specimen with a diameter of 20 mm and a thickness of 2 mm was prepared by high-temperature sintering. Scanning electron microscopy was used to observe the surface morphologies of nano-Cu-ZnO and the specimens. The antibacterial effect of Escherichia coli (E. coli) was quantitatively studied by the plate colony counting method. The CCK-8 method was used to evaluate in vitro the cytotoxicity of the tested piece to mouse fibroblasts (L929). Live and dead cells were observed by fluorescence microscopy. The mechanical properties of modified IPS E. Max Ceram veneering porcelain were tested by a three-point bending strength test. Results Under the scanning electron microscope, Cu-ZnO appears with a block-like structure and can be seen dispersed in the veneering porcelain. When the nano Cu-ZnO loading was 1 wt%, 2 wt%, 3 wt%, and 4 wt%, the antibacterial rates of the specimens were 24.85%, 67.94%, 96.92%, and 99.99%, respectively, and the difference between the experimental groups and the control group was statistically significant (F = 23.308,P = 0.001). The relative growth rate of each group was greater than 80% after coculture with mouse fibroblast cells (L929) for 1 day and 3 days, and there was no significant difference between the groups. The morphology of L929 cells was normal after coculture for 24 hours. With the increase in the Cu-ZnO concentration, the flexural strength of the specimen exhibited an increasing trend followed by a decreasing trend. The bending strength of the specimen loaded with 3 wt% nano Cu-ZnO reached the maximum value (84.728 ± 6.82) MPa, and there was no statistically significant difference between groups (F = 0.633,P = 0.702). Conclusion The antibacterial rate of IPS E. max Ceram veneering porcelain loaded with 3 wt% nano Cu-ZnO was more than 96% against E. coli after high-temperature sintering at 750 ℃. The bending strength reached the maximum (84.728 ± 6.82) MPa, and there was no obvious cytotoxicity.

Objective: The refining parameters of isoflavones from Puerariae flos were optimized by using HPD-100 macroporous adsorption resin, and its in vitro lipid-lowering effects were evaluated. Methods: The optimized refine parameters of isoflavones from Puerariae flos were studied by macroporous adsorption resin, and the lipid-lowering effects of isoflavones from Puerariae flos were evaluated by pancreatic lipase enzyme activity, pancreatic cholesterol esterase enzyme activity and micellar cholesterol solubility. Results: Adsorption parameters: Puerariae flos isoflavones were adsorbed with concentration of 100 mg/mL, pH 6.0 and upper column volume of 20 mL. Desorption parameters: the adsorption resins were washed by 70% ethanol with pH 5.0 at flow rate of 2.0 mL/min. The purity of refined Puerariae flos isoflavones were 81.47%. The inhibition ratio on pancreatic lipase enzyme, pancreatic cholesterol esterase enzyme and micellar cholesterol solubility were 40.73%, 52.32% and 46.17%, respectively, and these were increased by 26.65%, 47.46% and 37.53% respectively, compared with not refining. Conclusion: Puerariae flos isoflavones showed lipid-lowering effects by inhibiting pancreatic lipase enzyme activity, pancreatic cholesterol esterase enzyme activity and micellar cholesterol solubility.

Alzheimer's disease is a neurodegenerative disorder characterized by a decline in cognitive function. The β-amyloid (Aβ) hypothesis suggests that Aβ peptides can spontaneously aggregate into β-fragment-containing oligomers and protofibrils, and this activation of the amyloid pathway alters Ca2+ signaling in neurons, leading to neurotoxicity and thus apoptosis of neuronal cells. In our study, a blood-brain barrier crossing flavonol glycoside hyperoside was identified with anti-Aβ aggregation, BACE inhibitory, and neuroprotective effect in cellular or APP/PSEN1 double transgenic Alzheimer's disease mice model. While our pharmacokinetic data confirmed that intranasal administration of hyperoside resulted in a higher bio-availability in mice brain, further in vivo studies revealed that it improved motor deficit, spatial memory and learning ability of APP/PSEN1 mice with reducing level of Aβ plaques and GFAP in the cortex and hippocampus. Bioinformatics, computational docking and in vitro assay results suggested that hyperoside bind to Aβ and interacted with ryanodine receptors, then regulated cellular apoptosis via endoplasmic reticulum-mitochondrial calcium (Ca2+) signaling pathway. Consistently, it was confirmed that hyperoside increased Bcl2, decreased Bax and cyto-c protein levels, and ameliorated neuronal cell death in both in vitro and in vivo model. By regulating Aβ-induced cell death via regulation on Ca2+ signaling cascade and mitochondrial membrane potential, our study suggested that hyperoside may work as a potential therapeutic agent or preventive remedy for Alzheimer's disease.

BackgroundThe incidence of magnet ingestion by children has recently increased in China. Magnet ingestion is associated with an extremely high risk of gastrointestinal damage because loops of bowel can become trapped and squeezed between multiple magnets in different locations. However, the lack of imaging sensitivity makes clinical decision-making difficult.ObjectiveThis study was conducted to investigate the performance of ultrasound in diagnosing gastrointestinal magnet ingestion in children.MethodsFrom April 2017 to February 2021, all children with a history of magnet ingestion or a diagnosis of gastrointestinal magnet as shown by x-ray or ultrasound in our hospital were included as study candidates. Patients who were lost to follow-up or had known malformations of the gastrointestinal tract were excluded. Eligible patients were those with surgical or endoscopic confirmation of gastrointestinal magnet, those who passed the magnet out of the alimentary tract without assistance, and those with confirmed absence of the magnet on abdominal x-ray examination after 1 month of conservative treatment. All eligible patients' ultrasound and x-ray examination data were evaluated. The sensitivity, specificity, and area under the curve (AUC) of ultrasound was calculated for diagnosing magnet ingestion, locating the magnet (stomach, small intestine, or colon), and confirming the phenomenon of wall entrapment.ResultsOf 112 patients, 107 had a magnetic foreign body and 5 did not. Magnets were correctly detected by ultrasound in 97 patients, with an observed sensitivity of 90.65% and specificity of 100%. Satisfactory sensitivity was obtained for ultrasound localization of gastric magnets (96.30%) and small intestinal magnets (100.00%), but sensitivity for ultrasound localization of colonic magnets was relatively poor (73.33%). The discrimination of wall entrapment by ultrasound was good (AUC = 0.93), with an observed sensitivity and specificity of 92.00% and 93.62%, respectively.ConclusionsUltrasound can be used to locate gastrointestinal magnets (in the stomach, small intestine, or colon) with good clinical efficacy in identifying wall entrapment.

Macrobrachium rosenbergii is one of the shellfish species with high aquaculture value due to its increasing market demand. However, the comparatively low production volume compared to demand coupled with the rapid decline of the natural environment, consequently, drives the potential depletion of the wild population. The decrease in water pH related to anthropogenic pollution is one of the most critical factors affecting the early life performances of M. rosenbergii. Therefore, this study was designed to examine the effect of low water pH on feeding, growth and development of M. rosenbergii early life stages. Experimental water pH was set as neutral (7.7 ± 0.4); mild-acidic (6.4 ± 0.5) and acidic (5.4 ± 0.2) with triplication at a stocking density of 2 larvae/L for 30 days. As expected, M. rosenbergii larvae were highly sensitive to acidic pH with no larvae survived beyond 48 h of exposure. Feeding, survival and growth of larvae were adversely affected by mild-acidic pH exposure as compared to neutral pH. Larvae exposed to mild-acidic water pH experienced a prolonged larval period and only metamorphosed to the post-larval stage at day-30. Whilst under neutral water pH, larval that metamorphosed to post-larval was first observed on day-23. The negative impact of decreased pH, even in mild-acidic pH exposure, on the feeding, survival, growth and development of M. rosenbergii larvae highlights the urgency of periodic pH monitoring during M. rosenbergii larviculture.

Abstract Background Schistosomiasis is very common in the southern part of the Yangtze River Basin in China. It is mainly manifested as appendicitis, ulcers, hematomas, and thickening of the intestinal tract. Schistosomiasis of the appendix is rare, mainly manifested as appendicitis, which is easy to be misdiagnosed. Case presentation Here we report a rare case of a Chinese female whose intestinal mass manifested as intestinal polyps and was eventually diagnosed pathologically as schistosomiasis infection (appendix schistosomiasis). So far, there are rare relevant cases reported. Conclusions Intestinal schistosomiasis is easily misdiagnosed, and appendix schistosomiasis is rare. The final diagnosis requires pathology, especially surgical pathology.

Because of its light weight, high strength, designable, composite materials are more and more widely used in the field of aerospace, especially in the field of general aviation. In this paper, the structural design and analysis of the composite wing (include wing beam, rib, skin, leading edge and trailing edge) of a four-seater electric aircraft were carried out. The finite element numerical analysis model of the composite wing was established, and the deformation and strain distribution of the composite wing under the limited load required by airworthiness regulations were calculated. Finally, the static test of the whole composite wing was carried out to verify the accuracy of the finite element results. The finite element analysis and testing results showed that the structural design of the composite wing of the electric aircraft meets the structural strength requirements specified in the airworthiness standard (part 23 of China civil aviation regulations: air-worthiness requirements for normal, practical, special effects and commuting aircraft: CCAR 23-R3)

Abstract Background Only a small number of patients have utilized the home-based end-of-life care service in Shanghai that has been offered since 2012. This study explores how home-based end-of-life care is delivered in community health service centers in Shanghai and examines the difficulties in the delivery of the care. Methods This was a qualitative study in which data were collected from interviews and analyzed using qualitative content analysis. Nineteen health care providers with experience in delivering home-based end-of-life care in 12 community health service centers were recruited. The interviews were conducted between August 2018 and February 2019. Results Four themes emerged from the interviews: (i) Patients under home-based end-of-life care: Patients receiving the care were cancer patients with less than 1 year of life expectancy. The criteria for patients were broad. (ii) Service structure: The service was delivered regularly by the physicians and nurses using the approaches of home visits and/or telephone follow-ups. (iii) Service process: The service consisted of multiple components, including monitoring the patient’s condition, managing the patient’s symptoms, giving daily care instructions, performing nursing procedures, and giving psychological support. However, most of the care focused on monitoring the patients and managing their physical discomfort. (iv) Difficulties in delivering care: Being unable to provide the service and feeling powerless when facing psycho-spiritual problems were the two major difficulties. Three factors contributed to the suspension of the service: The gap between the service and the needs of the patients, a lack of patients, and low work motivation. The demand that the truth be concealed from the families and their attitude of avoiding talking about death were the key factors of the failure of psycho-spiritual care. Conclusions Several issues should be addressed before the service can be further developed, including fully understanding the needs and preferences of local patients and their families, securing more financial support and a better supply of drugs, delivering better training for staff, and ensuring greater rewards for individuals and institutions providing the service.

The Amur ide (Leuciscus waleckii) is a fish in the Cyprinidae family. Compared with other Amur ide living in freshwater ecosystems, the Amur ide population in Lake Dali Nor of China is famous for its high tolerance to the alkaline conditions of 54 mM (pH 9.6). Yet, surprisingly, the ionoregulatory mechanism responsible for this remarkable alkaline adaptation remains unclear. Therefore, this study sought to investigate how bicarbonate affects the acid-base balancing and ionoregulatory responses of this animal. Here, using a comparative approach, the alkali form of Amur ide and its ancestral freshwater form living in other freshwater basins were each exposed to 50 mM (pH 9.59 ± 0.09), a level close to the alkalinity of Lake Dali Nor, and their physiological (AE1) adjustment of ions and acid-base regulation were investigated. This study highlighted differences in blood pH and serum ions (e.g., Na+, K+, Cl−, and Ca2+), Na+/K+ ATPase (NKA) activity and its mRNA level, and mRNA expression of gill transporters (Na+/H+ exchanger member 2 and/or 3, Na+/HCO3- cotransporter (NBC1), Cl−/HCO3- exchanger, Na+/Cl− cotransporter (NCC), Na+/K+/2Cl− (NKCC1), SLC26A5, and SLC26A6) for alkalinity adaptation between the two forms of Amur ide differing in alkalinity tolerance. Specifically, close relationships among the serum Na+ and mRNA levels of NCC, NKCC1, and NHE, and also NKA and NBC1, in addition to serum Cl− and bicarbonate transporters (e.g., SLC26A5 and SLC26A6), characterized the alkali form of Amur ide. We propose that this ecotype can ensure its transepithelial Cl− and Na+ uptake/base secretions are highly functional, by its basolateral NKA with NBC1 and apical ionic transporters, and especially NCC incorporated with other transporters (e.g., SLC26). This suggests an evolved strong ability to maintain an ion osmotic and acid-base balance for more effectively facilitating its adaptability to the high alkaline environment. This study provides new insights into the physiological responses of the alkaline form of the Amur ide fish for adapting to extreme alkaline conditions. This information could be used as a reference to cultivating alkaline-tolerant fish species in abandoned alkaline waters.

Dysregulation of protein posttranslational modification (PTM) can lead to a variety of pathological processes, such as abnormal sperm development, malignant tumorigenesis, depression, and aging process. SIRT7 is a NAD+-dependent protein deacetylase. Besides known deacetylation, SIRT7 may also have the capacity to remove other acylation. However, the roles of SIRT7-induced other deacylation in aging are still largely unknown. Here, we found that the expression of SIRT7 was significantly increased in senescent fibroblasts and aged tissues. Knockdown or overexpression of SIRT7 can inhibit or promote fibroblast senescence. Knockdown of SIRT7 led to increased pan-lysine crotonylation (Kcr) levels in senescent fibroblasts. Using modern mass spectrometry (MS) technology, we identified 5,149 Kcr sites across 1,541 proteins in senescent fibroblasts, and providing the largest crotonylome dataset to date in senescent cells. Specifically, among the identified proteins, we found SIRT7 decrotonylated PHF5A, an alternative splicing (AS) factor, at K25. Decrotonylation of PHF5A K25 contributed to decreased CDK2 expression by retained intron (RI)-induced abnormal AS, thereby accelerating fibroblast senescence, and supporting a key role of PHF5A K25 decrotonylation in aging. Collectively, our data revealed the molecular mechanism of SIRT7-induced k25 decrotonylation of PHF5A regulating aging and provide new ideas and molecular targets for drug intervention in cellular aging and the treatment of aging-related diseases, and indicating that protein crotonylation has important implications in the regulation of aging progress.

Leuciscus waleckii is a freshwater fish that is known to inhabit the Dali Nor Lake, Inner Mongolia, China. The water in this lake has an HCO3–/CO32– concentration of 54 mM (pH 9.6) and a salinity of 0.6‰. The physiological mechanisms that allow this fish to tolerate these saline/alkaline conditions have yet to be elucidated. Transcriptional component analysis has shown that the expression levels of a large number of genes involved in the pathways responsible for osmo-ionoregulation and arachidonic acid metabolism pathway expression change significantly (p < 0.05) during the regulation of acid–base balance under high alkaline stress. In this study, we investigated the role of long non-coding RNAs (lncRNAs) during adaptation to high alkaline conditions. Fish were challenged to an NaHCO3-adjusted alkalinity of 0 mM, 30 mM (pH 9.44 ± 0.08), and 50 mM (pH 9.55 ± 0.06) for 20 days in the laboratory. Gill and kidney tissues were then collected for high-throughput sequencing assays. A total of 159 million clean reads were obtained by high-throughput sequencing, and 41,248 lncRNA transcripts were identified. Of these, the mean number of exons and the mean length of the lncRNA transcripts were 4.8 and 2,079 bp, respectively. Based on the analysis of differential lncRNA transcript expression, a total of 5,244 and 6,571 lncRNA transcripts were found to be differentially expressed in the gills and kidneys, respectively. Results derived from Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the coding genes were correlated with the lncRNA expression profiles. GO analysis showed that many lncRNAs were enriched in the following processes: “transporter activity,” “response to stimulus,” and “binding.” KEGG analysis further revealed that metabolic pathways were significantly enriched. A random selection of 16 lncRNA transcripts was tested by RT-qPCR; these results were consistent with our sequencing results. We found that a large number of genes, with the same expression profiles as those with differentially expressed lncRNAs, were associated with the regulation of acid–base balance, ion transport, and the excretion of ammonia and nitrogen. Collectively, our data indicate that lncRNA-regulated gene expression plays an important role in the process of adaptation to high alkaline conditions in L. waleckii.

Exosomes are nano-extracellular vesicles with diameters ranging from 30 to 150 nm, which are secreted by the cell. With their role in drug cargo loading, exosomes have been applied to carry compounds across the blood–brain barrier in order to target the central nervous system (CNS). In this study, high-purity exosomes isolated by the ultra-high-speed separation method were applied as the natural compound carrier, with the loading efficiency confirmed by UHPLC-MS analysis. Through the optimization of various cargo loading methods using exosomes, this study compared the efficiency of different ways for the separation of exosomes and the exosome encapsulation of natural compounds with increasing molecular weights via extensive in vitro and in vivo efficacy studies. In a pharmacokinetic study, our data suggested that the efficiency of compound’s loading into exosomes is positively correlated to its molecular weight. However, with a molecular weight of greater than 1109 Da, the exosome-encapsulated natural compounds were not able to pass through the blood–brain barrier (BBB). In vitro cellular models confirmed that three of the selected exosome-encapsulated natural compounds—baicalin, hederagenin and neferine—could reduce the level of neurodegenerative disease mutant proteins—including huntingtin 74 (HTT74), P301L tau and A53T α-synuclein (A53T α-syn)—more effectively than the compounds alone. With the traditional pharmacological role of the herbal plant Nelumbo nucifera in mitigating anxiety, exosome-encapsulated-neferine was, for the first time, reported to improve the motor deficits of APP/PS1 (amyloid precursor protein/ presenilin1) double transgenic mice, and to reduce the level of β-amyloid (Aβ) in the brain when compared with the same concentration of neferine alone. With the current trend in advocating medicine–food homology and green healthcare, this study has provided a rationale from in vitro to in vivo for the encapsulation of natural compounds using exosomes for the targeting of BBB permeability and neurodegenerative diseases in the future.