Development of a stable genetic transformation system in Erigeron breviscapus: a case study with EbYUC2 in relation to leaf number and flowering time.

MAIN CONCLUSION: A stable genetic transformation system for Erigeron breviscapus was developed. We cloned the EbYUC2 gene and genetically transformed it into Arabidopsis thaliana and E. breviscapus. The leaf number, YUC2 gene expression, and the endogenous auxin content in transgenic plants were significantly increased. Erigeron breviscapus is a prescription drug for the clinical treatment of cardiovascular and cerebrovascular diseases. The rosette leaves have the highest content of the major active compound scutellarin and are an important component in the yield of E. breviscapus. However, little is known about the genes related to the leaf number and flowering time of E. breviscapus. In our previous study, we identified three candidate genes related to the leaf number and flowering of E. breviscapus by combining resequencing data and genome-wide association study (GWAS). However, their specific functions remain to be characterized. In this study, we cloned and transformed the previously identified full-length EbYUC2 gene into Arabidopsis thaliana, developed the first stable genetic transformation system for E. breviscapus, and obtained the transgenic plants overexpressing EbYUC2. Compared with wild-type plants, the transgenic plants showed a significant increase in the number of leaves, which was correlated with the increased expression of EbYUC2. Consistently, the endogenous auxin content, particularly indole-3-acetic acid, in transgenic plants was also significantly increased. These results suggest that EbYUC2 may control the leaf number by regulating auxin biosynthesis, thereby laying a foundation for revealing the molecular mechanism governing the leaf number and flowering time of E. breviscapus.

Contribution of platelets to disruption of the blood-brain barrier during arterial baroreflex dysfunction.

BACKGROUND: Arterial baroreflex dysfunction, like many other central nervous system disorders, involves disruption of the blood-brain barrier, but what causes such disruption in ABR dysfunction is unclear. Here we explored the potential role of platelets in this disruption. METHODS: ABR dysfunction was induced in rats using sinoaortic denervation, and the effects on integrity of the blood-brain barrier were explored based on leakage of Evans blue or FITC-dextran, while the effects on expression of CD40L in platelets and of key proteins in microvascular endothelial cells were explored using immunohistochemistry, western blotting and enzyme-linked immunosorbent assay. Similar experiments were carried out in rat brain microvascular endothelial cell line, which we exposed to platelets taken from rats with ABR dysfunction. RESULTS: Sinoaortic denervation permeabilized the blood-brain barrier and downregulated zonula occludens-1 and occludin in rat brain, while upregulating expression of CD40L on the surface of platelets and stimulating platelet aggregation. Similar effects of permeabilization and downregulation were observed in healthy rats that received platelets from animals with ABR dysfunction, and in rat brain microvascular endothelial cells, but only in the presence of lipopolysaccharide. These effects were associated with activation of NF-κB signaling and upregulation of matrix metalloprotease-9. These effects of platelets from animals with ABR dysfunction were partially blocked by neutralizing antibody against CD40L or the platelet inhibitor clopidogrel. CONCLUSION: During ABR dysfunction, platelets may disrupt the blood-brain barrier when CD40L on their surface activates NF-kB signaling within cerebral microvascular endothelial cells, leading to upregulation of matrix metalloprotease-9. Our findings imply that targeting CD40L may be effective against cerebral diseases involving ABR dysfunction.

A united framework modeling of spatial-temporal characteristics for county-level agricultural carbon emission with an application to Hunan in China.

Effectively tackling extreme climate change requires sound knowledge about carbon emissions and their driving forces. Currently, agricultural carbon emission assessment often deals with its inventory, efficiency, determinants, and response independently, which will leave out the complex interactions among its various components, thus there is a lack of comprehensive, scalable, comparable explanations for agricultural carbon emissions. Herein, we introduce an integrated agricultural carbon emission assessment framework (IEDR): Inventory (I) × Efficiency (E) × Determinants (D) × Response (R), which was then applied to an illustration for the county-level agricultural carbon emissions in Hunan Province, China. Results show that: (1) Agricultural carbon emission inventory (ACEI) increased from 20.06 × 106 tC in 2006 to 21.99 × 106 tC in 2014 and decreased to 19.07 × 106 tC by 2020, depicting a fluctuating trend. Meanwhile, there was remarkable spatial heterogeneity, with higher ACEI in the North and South than in the East and West. (2) Agricultural carbon emission efficiency (ACEE) increased from 0.8520 in 2006 to 0.8992 in 2020, depicting a growing trend driven by technological progress. Spatially distributed in contrast to ACEI, regions with higher ACEE were located in the eastern and western areas. (3) ACEI was negatively correlated with ACEE (-0.657), indicating that increasing ACEE is a key strategy for reducing emissions. (4) The natural environment, rural development level, and policy support had critical impacts on ACEE and ACEI. In particular, the cultivated area and rural water affairs development were significant influences on ACEE and ACEI. Given the externalities of carbon emissions and its important public goods characteristics of the atmosphere, local carbon issues are also global concerns. Therefore, the case study of the IEDR model not only validates this theoretical paradigm and realizes regional responsibility for global carbon reduction but also supports and expands the theoretical and empirical corpus in the field of agricultural carbon emissions and efficiency, providing insights and references for other global regions facing similar challenges.

A geopolymer membrane for application in a structural mechanics and energy storage difunctional supercapacitor.

A structural mechanics and energy storage difunctional supercapacitor based on a geopolymer membrane injected with a 0.5 M Na2SO4 electrolyte and a pseudocapacitive electrode Mn7O13 is designed and assembled. The geopolymer membrane is prepared as a structural electrolyte with metakaolin and alkaline activator solution. The wide channels in the geopolymer matrix provide paths for ion movement. The Mn7O13 electrode is prepared by different hydrothermal treatments at different temperatures and times, and assembled with activated carbon and a geopolymer with different moduli to form a difunctional supercapacitor. The results show that the electrode sample annealed at 300 °C for 45 min after hydrothermal treatment at 160 °C for 24 h exhibits the best comprehensive performance. The specific capacitance of the electrode is 175.5 F g-1 (2392.6 F m-2) at 1 A g-1, and the specific capacitance of the difunctional structure supercapacitor assembled with a geopolymer with a modulus of 1.2 and cured for 28 days is 144.12 F g-1 (1960.0F m-2) at 1 A g-1 under 15 MPa.

Transcriptome analyses reveal the effects of mixed saline-alkali stress on indoleacetic acid and cytokinins in Malus hupehensis Rehd. leaves.

Saline-alkali stress is a universal abiotic stress factor limiting fruit tree cultivation worldwide. Apple (Malus×domestica Borkh.) is one of the fruits with the largest yields worldwide. Tea crabapple (Malus hupehensis Rehd. var. pingyiensis Jiang) is a type of common apple rootstock in China. Because facultative apomixis occurs in this species, it is often used in molecular research. The present study investigated the molecular mechanism of the response of indoleacetic acid (IAA) and cytokinins [zeatin, trans-zeatin riboside (tZR), isopentenyladenine (iP), and isopentenyladenosine (iPA)] to mixed saline-alkali stress (MSAS) in tea crabapple leaves. The endogenous hormone content of tea crabapple leaves under MSAS was measured, and the expression of stress response-related genes was analyzed by RNA sequencing. The results showed that the concentration of IAA was initially higher and then lower than that in the control, whereas the concentration of zeatin, tZR, iP, and iPA was higher than that in the control. A total of 1262 differentially expressed genes were identified in the three comparison groups. Further analyses suggested that IAA and cytokinin biosynthetic genes were mostly upregulated in tea crabapple leaves, indicating that auxin and cytokinin signaling pathway regulation occurred in response to MSAS. These findings suggest that IAA and cytokinins play an important role in the response of tea crabapple to MSAS. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01275-4.

Flow Electrode Capacitive Deionization System with Simultaneous Desalting of Na+ and Gathering of Na.

Oceans contain many freshwater resources and metal elements that people need, so the rational development of marine resources can solve the two major problems of shortage of freshwater resources and metal elements for people. To solve these two challenges, a system was designed to obtain freshwater resources and metallic elements simultaneously. An ion enrichment module was added to the conventional flow capacitor deionization system to collect metal elements while the seawater was deionized. A flowing electrode allows the metal elements to enter the flowing electrode through the desalination ability. It transports the metal elements to the enrichment module through the fluidity of the fluid while reducing the ion concentration at the flowing electrode, thus reducing the effect caused by the rejection of the same ion and collecting and enriching the metal elements. We purchased activated carbon to test the feasibility of the system with different mass fractions of activated carbon suspensions. The results showed that the elemental enrichment capacity of the system increased from 12.291 to 14.795 mg, and the enrichment rate increased from 13.536 to 16.294 mg cm-2 h-1 as the mass fraction of activated carbon increased. Thus, the system accomplished the goals of desalination and metal collection simultaneously.

Embryo morphologic quality in relation to the metabolic and cognitive development of singletons conceived by in vitro fertilization and intracytoplasmic sperm injection: a matched cohort study.

BACKGROUND: Embryos with higher morphologic quality grading may have a greater potential to achieve clinical pregnancy that leads to a live birth regardless of the type of cleavage-stage embryos or blastocysts. Few studies have investigated the impacts of embryo grading on the long-term health of the offspring. OBJECTIVE: This pilot study aimed to examine the associations between embryo morphologic quality and the physical, metabolic, and cognitive development of singletons conceived by in vitro fertilization and intracytoplasmic sperm injection at preschool age. STUDY DESIGN: This matched cohort study included singletons born to infertile couples who underwent fresh cleavage-stage embryo transfer cycles with good- or poor-quality embryos from 2014 to 2016 at the reproductive center of the Women's Hospital, School of Medicine, Zhejiang University. A total of 144 children, aged 4 to 6 years, participated in the follow-up assessment from 2020 to 2021, and the response rate of poor-quality embryo offspring was 39%. Singletons in the good-quality embryo group were matched with singletons in the poor-quality embryo group at a 2:1 ratio according to the fertilization method and the children's age (±1 year). We measured the offspring's height, weight, body mass index, blood pressure, thyroid hormone levels, and metabolic indicators. Neurodevelopmental assessments were performed using the Chinese version of the Wechsler Preschool and Primary Scale of Intelligence, Fourth Edition, and the Adaptive Behavior Assessment System, Second Edition. We also collected data from the medical records. A linear regression model was used to analyze the association between embryo morphologic quality and offspring health outcomes. RESULTS: A total of 48 singletons conceived with poor-quality embryo transfer and 96 matched singletons conceived with good-quality embryo transfer were included in the final analysis. Age, sex, height, weight, body mass index, blood pressure, thyroid function, and metabolic indicators were comparable between the 2 groups. After adjustment for potential risk factors by linear regression model 1 and model 2, poor-quality embryo offspring exhibited a tendency toward higher free thyroxine levels than offspring of good-quality embryo transfers (beta, 0.22; 95% confidence interval, 0.09-0.90; beta, 0.22; 95% confidence interval, 0.09-0.91, respectively), but this difference was not clinically significant. Regarding neurodevelopmental assessments, there was no difference in the full-scale intelligence quotient based on the Wechsler Preschool and Primary Scale of Intelligence (109.96±12.42 vs 109.60±14.46; P=.88) or the general adaptive index based on the Adaptive Behavior Assessment System (108.26±11.70 vs 108.08±13.44; P=.94) between the 2 groups. The subindices of the 2 tests were also comparable. These findings remained after linear regression analysis. CONCLUSION: At 4 to 6 years of age, singletons born from poor-quality embryo transfers have comparable metabolic and cognitive development as those born from good-quality embryo transfers using fresh cleavage-stage embryos. The results of this pilot study indicate that poor-quality embryos that can survive implantation and end in live birth are likely to have a developmental potential comparable to that of good-quality embryos.

Bilateral lung transplantation for Castleman disease with end-stage bronchiolitis obliterans.

Bronchiolitis obliterans (BO) is a severe complication of Castleman disease (CD), a rare lymphoproliferative disease with unclear pathogenesis. Currently, there are no reports on the safety or outcomes of bilateral lung transplantation in patients with BO due to CD. This study aimed to characterize the clinical manifestations and features of BO and CD. We retrospectively analyzed the medical records of six consecutive patients with BO and CD who underwent bilateral lung transplantation between December 2012 and December 2020. The average age of patients at lung transplantation was 33 ± 15 years, and the age range of patients at diagnosis of CD was about 9-56 years. The body mass index was 15.2 ± 1.9 kg/m2 . The average time from diagnosis to lung transplantation was 4.1 ± 2.7 years. All the patients had unicentric CD (UCD); five had concomitant paraneoplastic pemphigus, and four received extracorporeal membrane oxygenation during surgery. The average hospital stay was 51 ± 53 days. Infection was the most common postoperative complication. CD did not recur in any of the patients. Thus, bilateral lung transplantation is a viable and safe treatment for selected patients with CD and BO, which can improve the quality of life and prolong survival.

Preoperative Thoracic CT Findings Associated With Postoperative Mechanical Ventilation in Patients Undergoing Major Abdominal or Pelvic Surgery: A Matched Case-Control Study.

BACKGROUND. Postoperative prolonged mechanical ventilation is associated with increased morbidity and mortality. Reliable predictors of the need for postoperative mechanical ventilation after abdominal or pelvic surgeries are lacking. OBJECTIVE. The purpose of this study was to explore associations between preoperative thoracic CT findings and the need for postoperative mechanical ventilation after major abdominal or pelvic surgeries. METHODS. This retrospective case-control study included patients who underwent abdominal or pelvic surgeries during the period from January 1, 2014, through December 31, 2018, and had undergone preoperative thoracic CT. Case patients were patients who required postoperative mechanical ventilation. Control patients and case patients were matched at a 3:1 ratio on the basis of age, sex, body mass index, chronic obstructive pulmonary disease, smoking status, and surgery type. Two radiologists (readers 1 and 2) reviewed the CT images. Findings were compared between groups. RESULTS. The study included 165 patients (70 women, 95 men; mean age, 67.0 ± 9.7 [SD] years; 42 case patients and 123 matched control patients). Bronchial wall thickening and pericardial effusion were more frequent in case patients than control patients for reader 2 (10% vs 2%, p = .03; 17% vs 5%, p = .01) but not for reader 1. Pulmonary artery diameter (mean ± SD) was greater in case patients than control patients for reader 2 (2.9 ± 0.5 cm vs 2.8 ± 0.5 cm, p = .045) but not reader 1. Right lung height was lower in case patients than control patients for reader 1 (18.4 ± 2.9 cm vs 19.9 ± 2.7 cm, p = .01) and reader 2 (18.3 ± 2.9 cm vs 19.8 ± 2.7 cm, p = .01). Left lung height was lower in case patients than control patients for reader 1 (19.5 ± 3.1 cm vs 21.1 ± 2.6 cm, p = .01) and reader 2 (19.6 ± 2.4 cm vs 20.9 ± 2.6 cm, p = .01). Anteroposterior (AP) chest diameter was greater for case patients than control patients for reader 1 (14.0 ± 2.3 cm vs 12.9 ± 3.7 cm, p = .02) and reader 2 (14.2 ± 2.2 cm vs 13.2 ± 3.6 cm, p = .04). In a multivariable regression model using pooled reader data, bronchial wall thickening exhibited an odds ratio (OR) of 4.6 (95% CI, 1.3-16.5; p = .02); pericardial effusion, an OR of 5.1 (95% CI, 1.7-15.5; p = .004); pulmonary artery diameter, an OR of 1.4 per 1-cm increase (95% CI, 0.7-3.0; p = .32); mean lung height, an OR of 0.8 per 1-cm increase (95% CI, 0.7-1.001; p = .05); and AP chest diameter, an OR of 1.2 per 1-cm increase (95% CI, 1.013-1.4; p = .03). CONCLUSION. CT features are associated with the need for postoperative mechanical ventilation after abdominal or pelvic surgery. CLINICAL IMPACT. Many patients undergo thoracic CT before abdominal or pelvic surgery; the CT findings may complement preoperative clinical risk factors.

Inland water bodies in China: Features discovered in the long-term satellite data.

Water bodies (WBs), such as lakes, ponds, and impoundments, provide essential ecosystem services for human society, yet their characteristics and changes over large areas remain elusive. Here we used unprecedented data layers derived from all Landsat images available between 1984 and 2015 to understand the overall characteristics and changes of WBs between 2 epochs (i.e., 1984 to 1999 and 2000 to 2015) in China. Results show that the abundance estimate of WBs greater than 1 km2 and the total WB surface area were 0.3 to 1.5 times and 0.2 to 0.5 times more than the previous estimates, respectively. The size-abundance and shoreline-area relationships of WBs in China conformed to the classic power scaling law, in contradiction to most previous studies. WB changes with various occurrence probabilities show widespread coexistence of disappearance of existent and emergence of new WBs across China driven primarily by human activities and climate change. Our results highlight the importance of using appropriate long-term satellite data to reveal the true properties and dynamics of WBs over large areas, which is essential for developing scaling theories and understanding the relative impacts of human activities and climate change on water resources in the world.

First Discovery of Beta-Sitosterol as a Novel Antiviral Agent against White Spot Syndrome Virus.

The outbreak of white spot syndrome (WSS) is a looming challenge, due to dramatic losses to the crustacean aquaculture industry. However, at present, there are no prophylactic or therapeutic means to control this infectious viral disease. Here, we screened fifteen medicinal plants for their inhibitory activity on the white spot syndrome virus (WSSV), using red swamp crayfish (Procambarus clarkii) as a model species. The results showed that the crude extracts of Pinellia ternata (Thunb.) Breit. had the highest inhibitory effect (91.59%, 100 mg/kg) on WSSV proliferation, and its main component, beta-sitosterol, showed a much higher activity (95.79%, 50 mg/kg). Further, beta-sitosterol potently reduced (p < 0.01) viral loads and viral gene transcription levels in a concentration-dependent fashion, and significantly promoted the survival rate of WSSV-challenged crayfish (57.14%, 50 mg/kg). The co-incubation assay indicated that beta-sitosterol did not influence the infectivity of WSSV particles. Both pre- and post-treatment of beta-sitosterol exerted a significant inhibitory effect (p < 0.01) on the viral load in vivo. Mechanistically, beta-sitosterol not only interfered with the expression of viral genes (immediate early gene 1, ie1; DNA polymerase, DNApol) that are important in initiating WSSV transcription, but it also attenuated the hijacking of innate immune signaling pathways (Toll, IMD, and JAK/STAT pathways) by viral genes to block WSSV replication. Moreover, the expression of several antiviral immune, antioxidant, pro-inflammatory, and apoptosis-related genes changed significantly in beta-sitosterol-treated crayfish. Beta-sitosterol is a potent WSSV inhibitor and has the potential to be developed as an effective anti-WSSV agent against a WSS outbreak in crustacean aquaculture.

[Overview of systematic reviews and Meta-analysis of Aidi Injection combined with chemotherapy in treatment of non-small cell lung cancer].

At present, many systematic reviews(SRs)/Meta-analysis of Aidi Injection combined with chemotherapy in the treatment of non-small cell lung cancer(NSCLC) have been published, and the effectiveness has been proved.However, the methodological quality and evidence quality of these SRs/Meta-analysis have not been evaluated, and their guiding role in the clinical practice needs to be further verified.In this study, SRs/Meta-analysis of Aidi Injection combined with chemotherapy in the treatment of NSCLC were assessed to provide evidence overview and basis for the application and decision-making of this drug in clinical practice.PubMed, Cochrane Library, EMbase, CNKI, VIP, Wanfang, and SinoMed databases were searched for research articles on SRs/Meta-analysis of Aidi Injection combined with chemotherapy in the treatment of NSCLC.The methodological quality and evidence quality of included 15 articles on SRs/Meta-analysis were evaluated by using the AMSTAR-2 and GRADE system.The results of SRs/Meta-analysis suggested that Aidi Injection combined with chemotherapy had certain advantages over chemotherapy alone in improving short-term efficacy, improving quality of life, and reducing leukopenia, thrombocytopenia, and the incidence of gastrointestinal adverse events.The results of the AMSTAR-2 checklist showed low quality for 11 SRs/Meta-analysis and extremely low quality for another four SRs/Meta-analysis.The top problems included failure to provide the preliminary protocol or guide, unreported funding sources, and non-assessed risk of bias in the included articles on the results.According to the results of the GRADE assessment, 32 of the 148 outcome indicators were of intermediate quality, 40 were of low quality, and 76 were of extremely low quality.The critical factor leading to the downgrade was the risk of bias, followed by imprecision and publication bias.Aidi Injection combined with chemotherapy in the treatment of NSCLC can enhance efficacy and reduce toxicity.However, due to the low methodological quality and evidence quality of the included research articles, the efficacy and safety of Aidi Injection combined with chemotherapy in the treatment of NSCLC still need to be further confirmed by high-quality studies.In the follow-up original research and SRs/Meta-analysis, the corresponding quality evaluation standards should be strictly followed to improve the quality of evidence.

Using nutrient-rich solutions and adding multiple cytoprotective agents as new strategies to develop lung preservation solutions.

Commonly, donor lungs are preserved with low-potassium dextran glucose solution at low temperature. We hypothesized that adding nutrients and/or cytoprotective agents to preservation solutions improves donor lung quality. Human lung epithelial cells and human pulmonary microvascular endothelial cells cultured at 37°C with serum containing medium were switched to designated testing solutions at 4°C with 50% O2 for different cold ischemic time, followed by switching back to serum containing culture medium at 37°C to simulate reperfusion. We found that bicarbonate buffer system should be avoided in preservation solution. When pH was maintained at physiological levels, cell culture media showed better cell survival than in low-potassium dextran glucose solution. Phosphate-buffered cell culture media were further improved by adding colloid dextran 40. When rat donor lungs were preserved at 4°C for 24 h, phosphate-buffered Roswell Park Memorial Institute-1640 medium [RPMI-1640(p)] plus dextran 40 or adding cytoprotective agents (alpha 1 antitrypsin, raffinose, and glutathione) to low-potassium dextran glucose solution prevented alveolar wall swelling, apoptosis, activation of endothelial cells, and cellular edema. Using nutrient-rich solution and/or adding multiple cytoprotective agents is a new direction for designing and developing organ preservation solutions. Cell culture model, as a screening tool, reduces the use of animals and provides potential underlying mechanisms.

Bio-inspired multimodal 3D endoscope for image-guided and robotic surgery.

Image-guided and robotic surgery based on endoscopic imaging technologies can enhance cancer treatment by ideally removing all cancerous tissue and avoiding iatrogenic damage to healthy tissue. Surgeons evaluate the tumor margins at the cost of impeding surgical workflow or working with dimmed surgical illumination, since current endoscopic imaging systems cannot simultaneous and real-time color and near-infrared (NIR) fluorescence imaging under normal surgical illumination. To overcome this problem, a bio-inspired multimodal 3D endoscope combining the excellent characteristics of human eyes and compound eyes of mantis shrimp is proposed. This 3D endoscope, which achieves simultaneous and real-time imaging of three-dimensional stereoscopic, color, and NIR fluorescence, consists of three parts: a broad-band binocular optical system like as human eye, an optical relay system, and a multiband sensor inspired by the mantis shrimp's compound eye. By introducing an optical relay system, the two sub-images after the broad-band binocular optical system can be projected onto one and the same multiband sensor. A series of experiments demonstrate that this bio-inspired multimodal 3D endoscope not only provides surgeons with real-time feedback on the location of tumor tissue and lymph nodes but also creates an immersive experience for surgeons without impeding surgical workflow. Its excellent characteristics and good scalability can promote the further development and application of image-guided and robotic surgery.

Production of a novel bispecific protein ULBP1×CD19-scFv targeting the NKG2D receptor and CD19 to promote the activation of NK cells.

Natural killer (NK) cells are potent cytotoxic effector cells of the innate immune system and play an important role in tumor immunosurveillance and control. NKG2D is an activating receptor of NK cells. The NKG2D receptor-ligand system has contributed to immune cells recognizing tumor cells and the tumor microenvironment. In order to stretch the application of NK cells on adoptive immunotherapy for B-cell malignancies, we designed and produced a novel bispecific ULBP1×CD19-scFv fusion protein, in which the extracellular domain of NKG2D ligand ULBP1 was fused to a single chain variable fragment (scFv) of anti-CD19. The vector expressing ULBP1×CD19-scFv protein was constructed and expressed in Pichia pastoris. Effects of medium composition, concentration of methanol as the inducer, induction time and broth content in shake flask on the expression of the recombinant protein were investigated. The results showed that the optimized conditions for ULBP1×CD19-scFv expression were 1% methanol induction for 96 h with 15% broth content. The secreted recombinant protein was purified using ammonium sulfate fractionation and Ni-NTA affinity chromatography and the purity is about 93%. The cytotoxicity of NK92-MI cells against CD19+ Raji cells was enhanced in the presence of purified ULBP1×CD19-scFv protein. These results indicated that ULBP1 could be used as an activating element of bispecific killer engagers (BiKEs) and Pichia pastoris yeast might be an alternative expression host for BiKEs production.

Ophiopogon japonicus inhibits white spot syndrome virus proliferation in vivo and enhances immune response in Chinese mitten crab Eriocheir sinensis.

White spot syndrome virus (WSSV) is a fatal pathogen threatening global crustacean industry with no commercially available drugs to control. Herbal medicines have been widely used to treat a number of viral infections, which could offer a rich reserve for antiviral drug discovery. Here, we evaluated the inhibition activities of 30 herbal medicines against WSSV in Chinese mitten crab Eriocheir sinensis. A WSSV infection model in E. sinensis was firstly established in order to determine the antiviral effects of the plant extracts and to explore the potential action mechanisms. Results showed that the highest anti-WSSV activity was obtained by the treatment of Ophiopogon japonicus extract (93.03%, 100 mg/kg). O. japonicus treatment decreased viral loads in a dose-dependent manner and significantly improved the survival of WSSV-challenged crabs. O. japonicus reduced the expression of vital genes in viral life cycle in vivo, particularly for the immediate-early stage gene ie1. Further results indicated that O. japonicus could repress the JAK-STAT signaling pathway to block ie1 transcription. Moreover, O. japonicus could modulate certain immune genes such as the myosin, toll-like receptor, crustin, and prophenoloxidase in the interactions between WSSV and crabs. The up-regulated expression of pro-autophagic factors (Gabarap and Atg7) and elevated levels of antioxidant enzymes (SOD, CAT and GSH) suggested that O. japonicus may induce autophagy and attenuate WSSV-induced oxidative stress. Taken together, O. japonicus could inhibit WSSV proliferation and improve the survival of WSSV-challenged crabs. Thus, O. japonicus may have the potential to be developed as a preventive or therapeutic agent against WSSV, and its effective compounds merit further isolation and identification.

The active fragments of ghrelin cross the blood-brain barrier and enter the brain to produce antinociceptive effects after systemic administration.

G (1-5)-NH2, G (1-7)-NH2, and G (1-9) are the active fragments of ghrelin. The aim of this study was to investigate the antinociceptive effects, their ability to cross the blood-brain barrier, and the receptor mechanism(s) of these fragments using the tail withdrawal test in male Kunming mice. The antinociceptive effects of these fragments (2, 6, 20, and 60 nmol/mouse) were tested at 5, 10, 20, 30, 40, 50, and 60 min after intravenous (i.v.) injection. These fragments induced dose- and time-related antinociceptive effects relative to saline. Using the near infrared fluorescence imaging experiments, our results showed that these fragments could cross the brain-blood barrier and enter the brain. The antinociceptive effects of these fragments were completely antagonized by naloxone (intracerebroventricular, i.c.v.); however, naloxone methiodide (intraperitoneal, i.p.), which is the peripheral restricted opioid receptor antagonist, did not antagonize these antinociceptive effects. Furthermore, the GHS-R1α antagonist [D-Lys3]-GHRP-6 (i.c.v.) completely antagonized these antinociceptive effects, too. These results suggested that these fragments induced antinociceptive effects through central opioid receptors and GHS-R1α. In conclusion, our studies indicated that these active fragments of ghrelin could cross the brain-blood barrier and enter the brain and induce antinociceptive effects through central opioid receptors and GHS-R1α after intravenous injection.

Evaluation of the antiviral activity of naringenin, a major constituent of Typha angustifolia, against white spot syndrome virus in crayfish Procambarus clarkii.

White spot syndrome virus (WSSV) is a serious pathogen threatening global crustacean aquaculture with no commercially available drugs. Herbal medicines widely used in antiviral research offer a rich reserve for drug discovery. Here, we investigated the inhibitory activity of 13 herbal medicines against WSSV in crayfish Procambarus clarkii and discovered that naringenin (NAR) has potent anti-WSSV activity. In the preliminary screening, the extracts of Typha angustifolia displayed the highest inhibitory activity on WSSV replication (84.62%, 100 mg/kg). Further, NAR, the main active compound of T. angustifolia, showed a much higher inhibition rate (92.85%, 50 mg/kg). NAR repressed WSSV proliferation followed a dose-dependent manner and significantly improved the survival of WSSV-challenged crayfish. Moreover, pre- or post-treatment of NAR displayed a comparable inhibition on the viral loads. NAR decreased the transcriptional levels of vital genes in viral life cycle, particularly for the immediately early-stage gene ie1. Further results showed that NAR could decrease the STAT gene expression to block ie1 transcription. Besides, NAR modulated immune-related gene Hsp70, antioxidant (cMnSOD, mMnSOD, CAT, GST), anti-inflammatory (COX-1, COX-2) and pro-apoptosis-related factors (Bax and BI-1) to inhibit WSSV replication. Overall, these results suggest that NAR may have the potential to be developed as preventive or therapeutic agent against WSSV.

Simultaneous detection of multiple bacterial and viral aquatic pathogens using a fluorogenic loop-mediated isothermal amplification-based dual-sample microfluidic chip.

Rapid and user-friendly diagnostic tests are necessary for early diagnosis and immediate detection of diseases, particularly for on-site screening of pathogenic microorganisms in aquaculture. In this study, we developed a dual-sample microfluidic chip integrated with a real-time fluorogenic loop-mediated isothermal amplification assay (dual-sample on-chip LAMP) to simultaneously detect 10 pathogenic microorganisms, that is Aeromonas hydrophila, Edwardsiella tarda, Vibrio harveyi, V. alginolyticus, V. anguillarum, V. parahaemolyticus, V. vulnificus, infectious hypodermal and haematopoietic necrosis virus, infectious spleen and kidney necrosis virus, and white spot syndrome virus. This on-chip LAMP provided a nearly automated protocol that can analyse two samples simultaneously, and the tests achieved limits of detection (LOD) ranging from 100 to 10-1  pg/µl for genomic DNA of tested bacteria and 10-4 to 10-5  pg/µl for recombinant plasmid DNA of tested viruses, with run times averaging less than 30 min. The coefficient of variation for the time-to-positive value was less than 10%, reflecting a robust reproducibility. The clinical sensitivity and specificity were 93.52% and 85.53%, respectively, compared to conventional microbiological or clinical methods. The on-chip LAMP assay provides an effective dual-sample and multiple pathogen analysis, and thus would be applicable to on-site detection and routine monitoring of multiple pathogens in aquaculture.

Quercetin inhibiting the PD-1/PD-L1 interaction for immune-enhancing cancer chemopreventive agent.

Targeting the PD-1/PD-L1 immune checkpoints has achieved significant positive results in the treatment of multiple cancers. Quercetin is one of the most abundant dietary flavonoids found in various vegetables and fruits, and has a wide range of biological activities including immunomodulation. Here we report that quercetin dihydrate was screened and shown to inhibit the PD-1/PD-L1 interaction. Treatment with quercetin dihydrate promoted the killing activity of T cells on MDA-MB-231 and NCI-H460 cancer cells. Experiments using the xenograft mouse model showed that the growth rate of tumor volumes and masses in the quercetin dihydrate-treated mice were decreased. Immunohistochemistry of the tumors showed that CD8, GZMB, and IFN-γ were increased in the quercetin dihydrate-treated mice. These results suggest that quercetin dihydrate attenuates the inhibitory effect of PD-L1 on T cells by inhibiting the PD-1/PD-L1 interaction, which has an exciting potential to be used as a cancer chemopreventive agent.