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Abstract Background Refractory Helicobacter pylori (H. pylori) infection inevitably increase the difficulty of drug selection. Here, we described our experience with the use of a novel tetravalent IgY against H. pylori for the treatment of patients with refractory H. pylori infection. Methods Patients were randomly assigned to receive the standard quadruple therapy (amoxicillin, clarithromycin, omeprazole and bismuth potassium citrate ) for 2 weeks or 250 mg of avian polyclonal IgY orally twice a day for 4 weeks. The binding efficacy of IgY to H. pylori antigens was detected by western blotting13. C-urea breath test was performed to evaluate the eradication therap’s efficacy. The side effects of IgY were evaluated via various routine tests. The questionnaire was used to gather clinical symptoms and adverse reactions. Results Western blot analysis showed that tetravalent IgY simultaneously bind to VacA, HpaA, CagA and UreB of H. pylori. Tetravalent IgY had an eradication rate of 50.74% in patients with refractory H. pylori and an inhibition rate of 50.04% against DOB (delta over baseline) of 13C-urea. The symptom relief rate was 61.76% in thirty-four patients with clinical symptoms, and no adverse reactions were observed during tetravalent IgY treatment period. Conclusions Polyclonal avian tetravalent IgY reduced H. pylori infection, and showed good efficacy and safety in the treatment of refractory H. pylori infection patients, which represented an effective therapeutic option of choice for patients with refractory H. pylori infection.

Abstract Photothermal therapy (PTT) is a cancer-targeted treatment approach.The occurrence of tumors may be related to microbial infections (Viruses, bacteria, fungi, etc.), which probably provokes anti-tumor immunity. However, T cells in the context of cancer become exhausted and dysfunctional. Galectin-9 (Gal-9) is highly expressed in normal tissues and associates with body immune tolerance, and was firstly evidenced with much higher expression on the primary solid tumors than CD80/86 (B7) and CD274 (PD-L1) here, which suggests that Gal-9 may be a key factor in inhibiting the anti-tumor immunity, and its receptor T cell immunoglobulin and mucin domain 3 (TIM-3) was discovered on the cytotoxic T lymphocytes (CTL) with high expression as well based on the single cell analysis. The immune checkpoint communications showed that the Gal-9/TIM-3 axis played the most vital role on negatively regulating the anti-tumor immunity of CTL for melanoma. Then, we used a novel transdermal photothermal nanosensitizer (FSGG) loading Gal-9 siRNA (FSGG/siGal-9) for knocking the tumor cells down Gal-9 to block the Gal-9/TIM-3 axis and prohibit CTL exhaustion synergizing PTT against melanoma, which evidenced good effects on inhibiting tumor growth and enhancing anti-tumor immunity, named “photothermal immunotherapy”. This paper provides a new perspective for tumor prevention and treatment.

Melanoma showcases a complex interplay of genetic alterations and intra- and inter-cellular morphological changes during metastatic transformation. While pivotal, the role of specific mutations in dictating these changes still needs to be fully elucidated. Telomerase promoter mutations (TERTp mutations) significantly influence melanoma's progression, invasiveness, and resistance to various emerging treatments, including chemical inhibitors, telomerase inhibitors, targeted therapy, and immunotherapies. We aim to understand the morphological and phenotypic implications of the two dominant monoallelic TERTp mutations, C228T and C250T, enriched in melanoma metastasis. We developed isogenic clonal cell lines containing the TERTp mutations and utilized dual-color expression reporters steered by the endogenous Telomerase promoter, giving us allelic resolution. This approach allowed us to monitor morpholomic variations induced by these mutations. TERTp mutation-bearing cells exhibited significant morpholome differences from their wild-type counterparts, with increased allele expression patterns, augmented wound-healing rates, and unique spatiotemporal dynamics. Notably, the C250T mutation exerted more pronounced changes in the morpholome than C228T, suggesting a differential role in metastatic potential. Our findings underscore the distinct influence of TERTp mutations on melanoma's cellular architecture and behavior. The C250T mutation may offer a unique morpholomic and systems-driven advantage for metastasis. These insights provide a foundational understanding of how a non-coding mutation in melanoma metastasis affects the system, manifesting in cellular morpholome.

CRISPR/Cas9 genome editing is a widely used tool for creating genetic knock-ins, which allow for endogenous tagging of genes. This is in contrast with random insertion using viral vectors, where expression of the inserted transgene changes the total copy number of a gene in a cell and does not reflect the endogenous chromatin environment or any trans-acting regulation experienced at a locus. There are very few protocols for endogenous fluorescent tagging in macrophages. Here, we describe a protocol to design and test CRISPR guide RNAs and donor plasmids, to transfect them into RAW 264.7 mouse macrophage-like cells using the Neon transfection system and to grow up clonal populations of cells containing the endogenous knock-in at various loci. We have used this protocol to create endogenous fluorescent knock-ins in at least six loci, including both endogenously tagging genes and inserting transgenes in the Rosa26 and Tigre safe harbor loci. This protocol uses circular plasmid DNA as the donor template and delivers the sgRNA and Cas9 as an all-in-one expression plasmid. We designed this protocol for fluorescent protein knock-ins; it is best used when positive clones can be identified by fluorescence. However, it may be possible to adapt the protocol for non-fluorescent knock-ins. This protocol allows for the fairly straightforward creation of clonal populations of macrophages with tags at the endogenous loci of genes. We also describe how to set up imaging experiments in 24-well plates to track fluorescence in the edited cells over time.Key features• CRISPR knock-in of fluorescent proteins in RAW 264.7 mouse macrophages at diverse genomic loci.• This protocol is optimized for the use of the Neon transfection system.• Includes instructions for growing up edited clonal populations from single cells with one single-cell sorting step and efficient growth in conditioned media after cell sorting.• Designed for knocking in fluorescent proteins and screening transfected cells byFACS, but modification for non-fluorescent knock-ins may be possible.Graphical overview

Background: Clinical trial is a crucial step in the development of a new therapy (e.g., medication) and is remarkably expensive and time-consuming. Forecasting the approval of clinical trials accurately would enable us to circumvent trials destined to fail, thereby allowing us to allocate more resources to therapies with better chances. However, existing approval prediction algorithms did not quantify the uncertainty and provide interpretability, limiting their usage in real-world clinical trial management. Methods: This paper quantifies uncertainty and improves interpretability in clinical trial approval predictions. We devised a selective classification approach and integrated it with the Hierarchical Interaction Network, the state-of-the-art clinical trial prediction model. Selective classification, encompassing a spectrum of methods for uncertainty quantification, empowers the model to withhold decision-making in the face of samples marked by ambiguity or low confidence. This approach not only amplifies the accuracy of predictions for the instances it chooses to classify but also notably enhances the model’s interpretability. Results: Comprehensive experiments demonstrate that incorporating uncertainty markedly enhances the model’s performance. Specifically, the proposed method achieved 32.37%, 21.43%, and 13.27% relative improvement on area under the precision–recall curve over the base model (Hierarchical Interaction Network) in phase I, II, and III trial approval predictions, respectively. For phase III trials, our method reaches 0.9022 area under the precision–recall curve scores. In addition, we show a case study of interpretability that helps domain experts to understand model’s outcome. The code is publicly available at https://github.com/Vincent-1125/Uncertainty-Quantification-on-Clinical-Trial-Outcome-Prediction. Conclusion: Our approach not only measures model uncertainty but also greatly improves interpretability and performance for clinical trial approval prediction.

The relative ease of genetic manipulation in S. cerevisiae is one of its greatest strengths as a model eukaryotic organism. Researchers have leveraged this quality of the budding yeast to study the effects of a variety of genetic perturbations, such as deletion or overexpression, in a high-throughput manner. This has been accomplished by producing a number of strain libraries that can contain hundreds or even thousands of distinct yeast strains with unique genetic alterations. While these strategies have led to enormous increases in our understanding of the functions and roles that genes play within cells, the techniques used to screen genetically modified libraries of yeast strains typically rely on plate or sequencing-based assays that make it difficult to analyze gene expression changes over time. Microfluidic devices, combined with fluorescence microscopy, can allow gene expression dynamics of different strains to be captured in a continuous culture environment; however, these approaches often have significantly lower throughput compared to traditional techniques. To address these limitations, we have developed a microfluidic platform that uses an array pinning robot to allow for up to 48 different yeast strains to be transferred onto a single device. Here, we detail a validated methodology for constructing and setting up this microfluidic device, starting with the photolithography steps for constructing the wafer, then the soft lithography steps for making polydimethylsiloxane (PDMS) microfluidic devices, and finally the robotic arraying of strains onto the device for experiments. We have applied this device for dynamic screens of a protein aggregation library; however, this methodology has the potential to enable complex and dynamic screens of yeast libraries for a wide range of applications.Key features• Major steps of this protocol require access to specialized equipment (i.e., microfabrication tools typically found in a cleanroom facility and an array pinning robot).• Construction of microfluidic devices with multiple different feature heights using photolithography and soft lithography with PDMS.• Robotic spotting of up to 48 different yeast strains onto microfluidic devices.

Objective To investigate the value of Hisense computer-assisted surgery (CAS) system in the surgical treatment of pediatric surgical diseases. Methods A retrospective analysis was performed for the clinical data of children who underwent surgical treatment in Department of Pediatric Surgery in our hospital from March 2016 to October 2022. Hisense CAS was used for three-dimensional reconstruction of lesions and surrounding tissue for all children before surgery, and the application and significance of Hisense CAS in the surgical treatment of pediatric surgical diseases were discussed by analyzing the 3D reconstructed images of thoracic, abdominal, and pelvic diseases, the simulated surgical resection volume of lesions, the feasibility of surgery in children, and follow-up results. Results After 3D reconstruction using Hisense CAS, the images clearly displayed the shape and branches of vessels around lesions, and the spatial anatomic relationship between the lesion and its surrounding vessels could be observed from any angle, thereby helping with the accurate localization and assessment of lesions. In this study, three-dimensional reconstruction was performed for 323 children before surgery, with 372 cases of reconstruction in total, and all 323 children underwent surgery according to the preoperative surgical plan. Among these 323 children, 287 were followed up after surgery and 36 were lost to follow-up, with a follow-up time of 9 (2, 16) months. Among the 287 children, 286 had no significant postoperative complications with good survival conditions, and 1 child with nephroblastoma had intestinal obstruction after surgery and achieved good survival conditions after the second surgery. Conclusion Hisense CAS can provide an accurate preoperative evaluation of tumors in parenchymal organs such as the liver and the kidney, assist in the design of optimal surgical plans, and help to achieve more accurate, safe, and effective surgery for infants and young children, which provides important guidance for precise implementation of surgical operation.

Abstract The persistence of tumor load in multiple myeloma (MM) lead to relapse in patients achieving complete remission (CR). Appropriate and effective methods of myeloma tumor load monitoring are important for guiding clinical management. This study aimed to clarify the value of microvesicles in monitoring MM tumor load. Microvesicles in bone marrow and peripheral blood were isolated by differential ultracentrifugation and detected by flow cytometry. Western blotting was applied to assess myosin light chain phosphorylation levels. Flow cytometry to detect Ps+CD41a−, Ps+CD41a−CD138+, Ps+CD41a−BCMA+ microvesicles from bone marrow can be used to predict myeloma burden, furthermore, Ps+CD41a− microvesicles may as a potential index to MRD test. Mechanistically, the releasing of microvesicles from MM cell was regulated by Pim-2 Kinase via Phosphorylation of MLC-2 protein.

It is of great importance to treat a bacterial-infected wound by a smart dressing capable of delivering antibiotics in a smart manner without causing drug resistance. The construction of smart release nanocontainers responsive to near-infrared (NIR) laser irradiation in an on-demand and stepwise way is a promising strategy for avoiding the emergence of multidrug-resistant bacteria. Here, we develop a hydrogel composite made of alginate and nanotubes with an efficient NIR-triggered release of rifampicin and outstanding antibacterial ability. This composite hydrogel is prepared through co-encapsulating antibacterial drug (rifampicin), NIR-absorbing dye (indocyanine green), and phase-change materials (a eutectic mixture of fatty acids) into halloysite nanotubes, followed by incorporation into alginate hydrogels, allowing the in-situ gelation at room temperature and maintaining the integrity of drug-loaded nanotubes. Among them, the eutectic mixture with a melting point of 39 °C serves as the biocompatible phase-change material to facilitate the NIR-triggered drug release. The resultant phase-change material gated-nanotubes exhibit a prominent photothermal efficiency with multistep drug release under laser irradiation. In an in vitro assay, composite hydrogel provides good antibacterial potency against Staphylococcus aureus, one of the most prevalent microorganisms of dangerous gas gangrene. A bacterial-infected rat full-thickness wound model demonstrates that the NIR-responsive composite hydrogel inhibits the bacteria colonization and suppresses the inflammatory response caused by bacteria, promoting angiogenesis and collagen deposition to accelerate wound regeneration. The NIR-responsive composite hydrogel has a great potential as an antibacterial wound dressing functionalized with controlled multistep treatment of the infected sites.

Abstract Background The consumption of conspecific eggs by neonate larvae is a common behavior in aphidophagous coccinellids. However, it is still unknown what far-reaching benefits can be brought to the artificial diet-reared cannibals and whether the age of eggs and/or ovipositing parents can affect the cannibals’ fitness. Here, the multicolored Asian ladybird beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) was used as a research model because it exhibits much stronger cannibalism behavior than other beetles. Nine types of eggs, varying according to parental age at oviposition and egg age per se, were respectively provided as foods for H. axyridis neonate larvae until they developed to 2nd instar. After that, the larvae were fed on an artificial diet and whether egg consumption by neonate larvae had far-reaching benefits was evaluated. Those larvae that directly fed on artificial diet since hatching were served as control. Results Compared with the control beetles, the cannibal beetles (with few exceptions) had significantly higher 2nd instar-adult survivals (61.1–75.7% vs. 34.3%) and greatly shortened 2nd instar-adult duration (22.7–24.6 days vs. 27.8 days). At adult emergence, the female cannibal beetles (with few exceptions) had significantly higher fresh weights than the control beetles (14.5–16.3 mg vs. 11.7 mg). Among the nine types of cannibals, non-significant difference was detected in almost all the measured developmental parameters. Conclusion Consumption of conspecific eggs by neonate larvae of H. axyridis had far-reaching benefits for their subsequent development on artificial diet, and neither the egg age nor parental age at oviposition affects the cannibals’ fitness. These findings provided new insights into the combined application of conspecific eggs and artificial diets in the mass rearing of coccinellids.

For several decades, aging in Saccharomyces cerevisiae has been studied in hopes of understanding its causes and identifying conserved pathways that also drive aging in multicellular eukaryotes. While the short lifespan and unicellular nature of budding yeast has allowed its aging process to be observed by dissecting mother cells away from daughter cells under a microscope, this technique does not allow continuous, high-resolution, and high-throughput studies to be performed. Here, we present a protocol for constructing microfluidic devices for studying yeast aging that are free from these limitations. Our approach uses multilayer photolithography and soft lithography with polydimethylsiloxane (PDMS) to construct microfluidic devices with distinct single-cell trapping regions as well as channels for supplying media and removing recently born daughter cells. By doing so, aging yeast cells can be imaged at scale for the entirety of their lifespans, and the dynamics of molecular processes within single cells can be simultaneously tracked using fluorescence microscopy.Key features• This protocol requires access to a photolithography lab in a cleanroom facility.• Photolithography process for patterning photoresist on silicon wafers with multiple different feature heights.• Soft lithography process for making PDMS microfluidic devices from silicon wafer templates.

Abstract Large deformability and high sensitivity is difficult to be realized simultaneously in flexible sensors. Herein, taking advantage of the high permittivity and highly active surfaces of the ultrasmall barium titanate nanoparticles (BT NPs) and the high stretchability of the p(BA‐GMA) elastomer matrix, we propose a high‐performance soft stretchable sensor. The addition of the ultrasmall BT NPs can not only increase the permittivity and capacitance of polyacrylate‐matrix composite dielectric material to obtain a high sensitivity, but also basically maintains the excellent mechanical properties of the polymer matrix. The dielectric constants of the composite films increase from 5.68 to 13.13 at 10 kHz with the increase of BT NPs content from 0 to 15 vol.%, which results in a high capacitance of 236.16 pF for 15 vol.% BT/p(BA‐GMA) sensor. Combining the high permittivity and the large deformability (a maximal deformation of 87.2%), the 15 vol.% BT/p(BA‐GMA) sensor has high sensitivity and shows high linearity and stable output even if under dynamic measurement. The dual‐mode sensor that utilizes the orthogonality of capacitance‐resistance is designed, which shows excellent performance in monitoring human body movements and noncontact measurement. The results present that the BT/p(BA‐GMA)‐based sensor has high stability and reliability not exceed 65°C, which can meet the application requirements in dynamic monitoring.

Abstract The aim of this study was to investigate the characteristics of a novel type C lectin from Trichinella spiralis (TsCTL) and its role in larval invasion of intestinal epithelial cells (IECs). TsCTL has a carbohydrate recognition domain (CRD) of C-type lectin. The full-length TsCTL cDNA sequence was cloned and expressed in Escherichia coli BL21. The results of qPCR, Western blotting and immunofluorescence assays (IFAs) showed that TsCTL was a surface and secretory protein that was highly expressed at the T. spiralis intestinal infective larva (IIL) stages and primarily located at the cuticle, stichosome and embryos of the parasite. rTsCTL could specifically bind with IECs, and the binding site was localized in the IEC nucleus and cytoplasm. The IFA results showed that natural TsCTL was secreted and bound to the enteral epithelium at the intestinal stage of T. spiralis infection. The rTsCTL had a haemagglutinating effect on murine erythrocytes, while mannose was able to inhibit the rTsCTL agglutinating effect for mouse erythrocytes. rTsCTL accelerated larval intrusion into the IECs, whereas anti-rTsCTL antibodies and mannose significantly impeded larval intrusion in a dose-dependent manner. The results indicated that TsCTL specifically binds to IECs and promotes larval invasion of intestinal epithelium, and it might be a potential target of vaccines against T. spiralis enteral stages.

Abstract Background As the most common malignant tumor of primary renal tumor, renal cell carcinoma (RCC) is the highly invasive disease with high mortality. AKT is a serine/threonine kinase that play a critical role in the phosphoinositide 3-kinase (PI3K) signaling pathway, and it is an attractive target for RCC treatment. The aim of present study was to investigate the effect of AKT silence on malignant behavior of renal cell carcinoma cells. Methods AKT expression was quantified by immunohistochemistry in tumor tissues and normal tissues. The human RCC cell lines Caki-2 cell were chosen for this study. The optimal silencing siRNA was subsequently selected by RT-qPCR and western blot. The effect of AKT silence on RCC cells was investigated by CCK8 assay, transwell assay, scratch test and flow cytometry. The AKT1 expression in human renal cell carcinoma tissue was detected by immunohistochemical staining. Results The AKT in Caki-2 cells was silenced successfully. The results shown AKT silence could inhibit cell proliferation, invasion, and, migration. In addition, AKT silence could promote Caki-2 cell apoptosis with prevention of RCC cells move from G1 phase to S phase. Immunohistochemical staining revealed significant difference of expression of AKT1 in RCC tissues and normal renal tissues. Taken together, AKT family members might involve in malignant growth of RCC, and might be a potential therapeutic target. Conclusion Our data show that AKT silence inhibited cell proliferation, invasion, and, migration of Caki-2 cell, and promoted Caki-2 cell apoptosis. Moreover, AKT silence prevented RCC cells move from G1 phase to S phase. Therefore, AKT may act as an effective therapeutic target for RCC.

Abstract Background Postoperative cognitive dysfunction (POCD) are a common complication of the central nervous system following surgery and anesthesia. The specific pathogenesis and effective therapeutics of POCD need to be further studied. Ginkgolide B (GB), a platelet-activating factor receptor-specific antagonist, has been suggested to have strong anti-inflammatory effects. Here we tested the effects and mechanism of GB on POCD of aged rats. Methods Neurobehavioral tests were used to investigate the effect of GB pretreatment on POCD. The hippocampus were harvested to test the expression of proinflammatory cytokines by ELISA. The expression of the microglial marker ionized calcium-binding adaptor molecule-1 (Iba-1) in the hippocampus was evaluated by western blot assay and immunohistochemistry. A Nissl staining experiment was used to detect the neuronal numbers in the hippocampus. Results Surgery might result in the overexpression of platelet activating factor (PAF) in the plasma and hippocampus and might cause hippocampus-dependent memory impairment. GB pretreatment, inhibited the activation of microglia, reduced the levels of IL-1β and TNF-α, decreased the loss of neurons after surgery, and prevented POCD in aged rats. Conclusion Our findings suggested that PAF was involved in the development of POCD. Improvement of POCD by PAF antagonist GB was associated with the inhibition of microgliosis-mediated neuroinflammation and neuronal apoptosis in aged rats.

Objective: Single factor experiment and response surface methodology were used to optimize the ultrasonic extraction of oridonin from Rabdosiaru bescens and the antitumor effect in vitro was studied. Methods: Three key factors including solid-liquid ratio, ultrasonic time, ultrasonic power were selected by one-factor-at-a-time experiments. HepG2 cells were cultured in vitro. The antitumor activity of oridonin was investigated by studying the survival rate of HepG2 cells and the expression of apoptotic proteins caspase-3 and caspase-9. Results: The optimum conditions of ultrasonic extraction of oridonin from Rabdosiaru bescens were as follows: Solid-liquid ratio was 1:18 g/mL, ultrasonic time was 1.5 h at 190 W ultrasonic power, the extraction yield of oridonin reached 4.122%±0.102%. In vitro, oridonin could significantly reduced the survival rate of HepG2 cells and increased the expression of caspase-3 and caspase-9. And when the concentration of oridonin exceeds 8 μmol/L, the survival rate and apoptotic protein of hepatoma cells were significantly affected (P

Abstract Background Laminins are high-molecular weight (400 ~ 900 kDa) proteins in extracellular matrix, which serve as major component of the basal lamina, and play a crucial role in promoting tumor cell migration. This study aimed at characterizing the role of laminin in promoting cancer development, and elucidating the mechanism of tumor progression driven by laminin-Notch signaling in bladder cancer. Methods 2D collagen/laminin culture system was established and CCK-8/transwell assay was conducted to evaluate the proliferation/migration ability of Biu-87 and MB49 cells cultured on 2D gels. Activation of integrins-Notch1 signaling was determined by western blotting. Orthotopic bladder cancer mice model was established to assess the therapeutic effects of Notch inhibitor. Results Our study demonstrated that extracellular laminin can trigger tumor cell proliferation/migration through integrin α6β4/Notch1 signaling in bladder cancer. Inhibition of Telomere repeat-binding factor 3 (TRB3)/Jagged Canonical Notch Ligand 1 (JAG1) signaling suppressed Notch signals activation induced by laminin-integrin axis. In MB49 orthotopic bladder cancer mice model, Notch inhibitor SAHM1 efficiently improved tumor suppressive effects of chemotherapy and prolonged survival time of tumor-bearing mice. Conclusion In conclusion, we show that, in bladder cancer, extracellular laminin induced the activation of Notch pathway through integrin α6β4/TRB3/JAG3, and disclosed a novel role of laminin in bladder cancer cells proliferation or migration.

Summary: The repetitive applications of vaccine boosters have been brought up in face of continuous emergence of SARS-CoV-2 variants with neutralization escape mutations, but their protective efficacy and potential adverse effects remain largely unknown. Here, we compared the humoral and cellular immune responses of an extended course of recombinant receptor binding domain (RBD) vaccine boosters with those from conventional immunization strategy in a Balb/c mice model. Multiple vaccine boosters after the conventional vaccination course significantly decreased RBD-specific antibody titers and serum neutralizing efficacy against the Delta and Omicron variants, and profoundly impaired CD4+ and CD8+T cell activation and increased PD-1 and LAG-3 expressions in these T cells. Mechanistically, we confirmed that extended vaccination with RBD boosters overturned the protective immune memories by promoting adaptive immune tolerance. Our findings demonstrate potential risks with the continuous use of SARS-CoV-2 vaccine boosters, providing immediate implications for the global COVID-19 vaccination enhancement strategies.

Study region: The Shihmen reservoir, the second largest reservoir in northern Taiwan, is flooded frequently, and featured by short period of flood peak due to uneven distribution of rainfall and mountainous topography. Study focus: We proposed a novel streamflow-oriented ensemble recurrent neural networks (ERNN) method to merge three precipitation products, namely gauge measurements, radar and satellite rainfall products. We analyzed (1) the effect of artificial intelligence method for bias correction of precipitation products with reference to streamflow, (2) the comparison of two widely used arithmetic average (AA) and Bayesian model averaging (BMA) methods with ERNN, (3) the performance of merged rainfalls and the original three precipitation products in inflow forecasting during 13 typhoon events. New hydrological insights: We found that all precipitation products are biased and can be appropriately adjusted with an improvement in inflow forecasting of about 2.5 %, 21.8 % and 60.7 % for gauge, radar and satellite rainfall products in terms of RMSE. After rainfall merging, RMSE for radar and satellite are further improved by 14% and 36% at least. ERNN merging method indicates that the optimal merging weights for gauge, radar and satellite are within the range of 0.52–0.56, 0.35–0.39 and 0.08–0.09, respectively according to 95 % confidence interval. The merged rainfall product skillfully predicts the inflow with a lead time of five hours, and ERNN merging method is superior to traditional AA and BMA methods, with NS up to 0.85 and CRPS reduced by near 50 % compared to BMA. Additionally, ERNN is found to capture the peak times and peak flows with the smallest error. Overall, this study provides a potential approach to merge multiple rainfall products and to obtain the effective rainfall by fitting the hydrological responses over mountainous watersheds, where observational biases may frequently occur in gauge, radar and satellite measurements.

Genetic heterogeneity of metastatic dissemination has proven challenging to identify exploitable markers of metastasis; this bottom-up approach has caused a stalemate between advances in metastasis and the late stage of the disease. Advancements in quantitative cellular imaging have allowed the detection of morphological phenotype changes specific to metastasis, the morphological changes connected to the underlying complex signaling pathways, and a robust readout of metastatic cell state. This review focuses on the recent machine and deep learning developments to gain detailed information about the metastatic cell state using light microscopy. We describe the latest studies using quantitative cell imaging approaches to identify cell appearance-based metastatic patterns. We discuss how quantitative cancer biologists can use these frameworks to work backward toward exploitable hidden drivers in the metastatic cascade and pioneering new Frontier drug discoveries specific for metastasis.

Chromatin instability and protein homeostasis (proteostasis) stress are two well-established hallmarks of aging, which have been considered largely independent of each other. Using microfluidics and single-cell imaging approaches, we observed that, during the replicative aging of Saccharomyces cerevisiae, a challenge to proteostasis occurs specifically in the fraction of cells with decreased stability within the ribosomal DNA (rDNA). A screen of 170 yeast RNA-binding proteins identified ribosomal RNA (rRNA)-binding proteins as the most enriched group that aggregate upon a decrease in rDNA stability induced by inhibition of a conserved lysine deacetylase Sir2. Further, loss of rDNA stability induces age-dependent aggregation of rRNA-binding proteins through aberrant overproduction of rRNAs. These aggregates contribute to age-induced proteostasis decline and limit cellular lifespan. Our findings reveal a mechanism underlying the interconnection between chromatin instability and proteostasis stress and highlight the importance of cell-to-cell variability in aging processes.

Abstract Background Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) is an important biocontrol agent in native region of Asia, while its high propensity for cannibalism causes great obstacles in mass production. Provisioning obstructs in rearing containers could decrease the cannibalism of ladybird beetles. In this study, three different patterns of obstructs (Con-A, Con-B, and Con-C) were designed and equipped in plastic Petri dishes (95.38 cm3) as rearing units, and their efficiency for H. axyridis larval survival was tested. The potential of the high-density rearing was then evaluated using the optimal units with 16, 24, or 32 larvae per unit (named 16-L, 24-L, or 32-L, respectively). Results Larval survivals in obstructs-equipped units were generally higher than those in control, and significantly increased with the Con-C units (39.8% versus 74.2% at adult stage). With the Con-C units, the survivals were significantly higher at 16-L density (82.5%) than those at 24-L density (62.5%), but both were non-significantly different from those at 32-L density (70.0%). The weights of newly emerged adults (fit with the expected sex ratio of 1:1) at the higher densities were lower than those at 16-L density. Conclusions The results demonstrate that H. axyridis can be reared at a higher density (≈ 0.336 larvae/cm3) in a constrained unit and highlight the effects of obstructs in reducing cannibalism and improving insect survivals.

Abstract Inorganic pyrophosphatase (PPase) participates in energy cycle and plays a vital role in hydrolysis of inorganic pyrophosphate (PPi) into inorganic phosphate (Pi). The aim of this study was to investigate the biological properties of a Trichinella spiralis PPase (TsPPase) and its role in larval molting and developmental process. The predicted TsPPase consisted of 367 amino acids with a molecular mass of 41.48 kDa and a pI of 5.76. Amino acid sequence alignment and phylogenetic analysis showed that the TsPPase gene encodes a functional family I soluble PPase with the same characteristics as prokaryotic, plant and animal/fungal soluble PPase. The rTsPPase was expressed and purified, it has the activity to catalyze the hydrolysis of PPi to Pi, and the activity was dependent on Mg2+, pH and temperature. The enzymatic activity of rTsPPase was significantly inhibited after its metal binding sites mutation. TsPPase was transcribed and expressed in all T. spiralis phases, especially in muscle larvae (ML) and intestinal infective larvae (IIL). Immunofluorescence assay (IFA) revealed that TsPPase was mainly located in cuticle and stichosome. When the ML and IIL were treated with TsPPase-specific siRNA-279, TsPPase expression and enzymatic activity were obviously reduced, the larval molting and development were also impeded. Intestinal IIL as well as AW burden, IIL molting rates from mice infected with siRNA-treated ML were obviously suppressed. The results indicated that rTsPPase possesses the enzymatic activity of native inorganic pyrophosphatase, and TsPPase plays an important role in development and molting process of intestinal T. spiralis larval stages.

In this paper, we present long term observations of atmospheric nitrogen dioxide (NO2) and formaldehyde (HCHO) in Nanjing using a Multi-AXis Differential Optical Absorption Spectroscopy (MAX-DOAS) instrument. Ground based MAX-DOAS measurements were performed from April 2013 to February 2017. The MAX-DOAS measurements of NO2 and HCHO vertical column densities (VCDs) are used to validate OMI satellite observations over Nanjing. The comparison shows that the OMI observations of NO2 correlate well with the MAX-DOAS data with Pearson correlation coefficient (R) of 0.91. The comparison result of MAX-DOAS and OMI observations of HCHO VCD shows a good agreement with R of 0.75 and the slope of the regression line is 0.99. The age weighted backward propagation approach is applied to the MAX-DOAS measurements of NO2 and HCHO to reconstruct the spatial distribution of NO2 and HCHO over the Yangtze River Delta during summer and winter time. The reconstructed NO2 fields show a distinct agreement with OMI satellite observations. However, due to the short atmospheric lifetime of HCHO, the backward propagated HCHO data does not show a strong spatial correlation with the OMI HCHO observations. The result shows the MAX-DOAS measurements are sensitive to the air pollution transportation in the Yangtze River Delta, indicating the air quality in Nanjing is significantly influenced by regional transportation of air pollutants.

Abstract Background ZKSCAN3, a zinc-finger transcription factor containing KRAB and SCAN domains, has been reported to be regulated in several human cancers. However, its expression and function in hepatocellular carcinoma (HCC) remains unknown. Methods Expression of ZKSCAN3 in HCC was analyzed by western blotting, immunohistochemistry, and real time PCR. Its correlation with the clinicopathological characteristics and prognosis of HCC patients was analyzed. The effects of ZKSCAN3 on the migration and invasion were determined by Transwell assays. The potential downstream targets of ZKSCAN3 and related molecular mechanisms were clarified by Western blot and dual luciferase reporter assay. Results In this study, we demonstrated for the first time that ZKSCAN3 mRNA and protein was up-regulated in HCC tissues and cell lines. High ZKSCAN3 expression was significantly associated with poor prognostic features, including advanced TNM stage and vascular invasion. For 5-year survival, ZKSCAN3 served as a potential prognostic marker of HCC patients. Functionally, ZKSCAN3 promoted migration, invasion and EMT progress via directly binding to integrin β4 (ITGB4) promoter and enhanced its expression. Further investigation proved that ITGB4 triggers the focal adhesion kinase (FAK) to activate the AKT signaling pathway. Inactivation of FAK and AKT by their specific inhibitors respectively reversed the effects of ZKSCAN3 on HCC cells. In addition, we demonstrated that ZKSCAN3 expression was regulated by miR-124. In HCC tissues. MiR-124 has an inverse correlation with ZKSCAN3 expression. Conclusion We demonstrate for the first time that ZKSCAN3 is overexpressed in HCC tissues and promotes migration, invasion and EMT process through ITGB4-dependent FAK/AKT activation, which was regulated by miR-124, suggesting the potential therapeutic value for HCC.

Aging is a complex, yet pervasive phenomenon in biology. As human cells steadily succumb to the deteriorating effects of aging, so too comes a host of age-related ailments such as neurodegenerative disorders, cardiovascular disease and cancer. Therefore, elucidation of the molecular networks that drive aging is of paramount importance to human health. Progress toward this goal has been aided by studies from simple model organisms such as Saccharomyces cerevisiae. While work in budding yeast has already revealed much about the basic biology of aging as well as a number of evolutionarily conserved pathways involved in this process, recent technological advances are poised to greatly expand our knowledge of aging in this simple eukaryote. Here, we review the latest developments in microfluidics, single-cell analysis and high-throughput technologies for studying single-cell replicative aging in S. cerevisiae. We detail the challenges each of these methods addresses as well as the unique insights into aging that each has provided. We conclude with a discussion of potential future applications of these techniques as well as the importance of single-cell dynamics and quantitative biology approaches for understanding cell aging.

Abstract An ultrasmall chemodynamic therapeutic (CDT) agent with favorable specificity to tumor microenvironment, i.e., high level of glutathione (GSH) and hydrogen peroxide (H2O2), is reported. The coordination polymer nanodot between divalent copper (Cu2+) and gallic acid (GA) is shortly named as Cu–GA. The ultrasmall size of Cu–GA (2.16 ± 0.3 nm) results in a large specific surface area, which is beneficial for improving its catalytic performance. After endocytosis into tumor cell interior, Cu–GA promotes GSH‐activated and H2O2‐reinforced CDT in situ, wherein divalent Cu(II) is reduced to monovalent Cu(I) by GSH and induced GSH depletion. Subsequently, the generated Cu(I) catalyzes local H2O2 to generate toxic hydroxyl radical (•OH) via Fenton‐like reaction, and •OH leads to tumor cell apoptosis. The higher levels of GSH and H2O2 in the tumor cell interior significantly improve the efficiency of CDT, and meanwhile protect the normal cells. In addition, the ultrasmall size of Cu–GA facilitates its fast clearance and eliminates long‐term body retention, with minimized systemic toxicity during the treatment in vivo. Therefore, as a novel copper‐based nanoformulation specifically responsive to the tumor microenvironment, Cu–GA provides promising potentials in CDT.

Background Trichinellosis is a serious zoonotic disease distributed around the world. It is needed to develop a safe, effective and feasible anti-Trichinella vaccine for prevention and control of trichinellosis. The aim of this study was to construct a recombinant Lactobacillus plantarum encoding Trichinella spiralis inorganic pyrophosphatase (TsPPase) and investigate its immune protective effects against T. spiralis infection. Methodology/Principal findings The growth of recombinant L. plantarum was not affected by TsPPase/pSIP409-pgsA′ plasmid, and the recombinant plasmid was inherited stably in bacteria. Western blot and immunofluorescence assay (IFA) indicated that the rTsPPase was expressed on the surface of recombinant L. plantarum. Oral vaccination with rTsPPase induced higher levels of specific serum IgG, IgG1, IgG2a and mucosal secretory IgA (sIgA) in BALB/c mice. ELISA analysis revealed that the levels of IFN-γ and IL-4 released from spleen, mesenteric lymph nodes and Peyer’s patches were evidently increased at 2–4 weeks following vaccination, compared to MRS (De Man, Rogosa, Sharpe) medium control group (P < 0.05). Immunization of mice with rTsPPase exhibited a 67.18, 54.78 and 51.91% reduction of intestinal infective larvae, adult worms and muscle larvae at 24 hours post infection (hpi), 6 days post infection (dpi) and 35 dpi, respectively (P < 0.05), and the larval molting and development was significantly inhibited by 45.45% at 24 hpi, compared to the MRS group. Conclusions TsPPase plays a crucial role in T. spiralis molting and development, oral vaccination with rTsPPase induced a significant local mucosal sIgA response and systemic Th1/Th2 immune response, and immune protection against T. spiralis infection in BALB/c mice. Author summary In the previous study, a Trichinella spiralis inorganic pyrophosphatase (TsPPase) was expressed and its role in larval molting and development was observed. In this study, a recombinant TsPPase/pSIP409-pgsA′ plasmid was constructed and transferred into Lactobacillus plantarum NC8, the rTsPPase was expressed on the surface of recombinant L. plantarum NC8. Oral immunization of mice with rTsPPase DNA vaccine elicited a high level of specific serum IgG, IgG1, IgG2a and mucosal secretory IgA (sIgA). The levels of IFN-γ and IL-4 released from spleen, mesenteric lymph nodes and Peyer’s patches were evidently increased at 2–4 weeks following vaccination. Immunization of mice with rTsPPase showed a significant reduction of intestinal infective larvae, adult worms and muscle larvae, and intestinal larval molting and development was significantly suppressed. The results indicated that oral vaccination with rTsPPase elicited a significant local mucosal sIgA response and specific systemic Th1/Th2 immune response, and an obvious protective immunity against T. spiralis infection.