Reconstruction of TNF-α with specific isoelectric point released from SPIONs basing on variable charge to enhance pH-sensitive controlled-release.

The clinical application of tumor necrosis factor-α (TNF-α) is limited by its short half-life, subeffective concentration in the targeted area and severe systemic toxicity. In this study, the recombinant polypeptide S4-TNF-α was constructed and coupled with chitosan-modified superparamagnetic iron oxide nanoparticles (S4-TNF-α-SPIONs) to achieve pH-sensitive controlled release and active tumor targeting activity. The isoelectric point (pI) of S4-TNF-α was reconstructed to approach the pH of the tumor microenvironment. The negative-charge S4-TNF-α was adsorbed to chitosan-modified superparamagnetic iron oxide nanoparticles (CS-SPIONs) with a positive charge through electrostatic adsorption at physiological pH. The acidic tumor microenvironment endowed S4-TNF-α with a zero charge, which accelerated S4-TNF-α release from CS-SPIONs. Our studies showed that S4-TNF-α-SPIONs displayed an ideal pH-sensitive controlled release capacity and improved antitumor effects. Our study presents a novel approach to enhance the pH-sensitive controlled-release of genetically engineered drugs by adjusting their pI to match the pH of the tumor microenvironment.

Controlled SPION-Exosomes Loaded with Quercetin Preserves Pancreatic Beta Cell Survival and Function in Type 2 Diabetes Mellitus.

Introduction: Quercetin has an ideal therapeutic effect on islet function improvement in type 2 diabetes mellitus (T2DM). However, the therapeutic benefit of quercetin is hindered by its poor bioavailability and limited concentration in pancreatic islets. In this study, superparamagnetic iron oxide nanoparticle (SPION)-modified exosomes were prepared to load quercetin, hoping to endow quercetin with enhanced water solubility and active targeting capacity with the help of magnetic force (MF). Methods: Transferrin-modified SPIONs (Tf-SPIONs) were synthesized by exploiting N-hydroxysuccinimidyl (NHS) conjugation chemistry, and quercetin-loaded exosomes (Qu-exosomes) were acquired by electroporation. Tf-SPION-modified quercetin-loaded exosomes (Qu-exosome-SPIONs) were generated by the self-assembly of transferrin (Tf) and the transferrin receptor (TfR). The solubility of quercetin was determined by high-performance liquid chromatography (HPLC) analysis. The pancreatic islet targeting capacity and insulin secretagogue and antiapoptotic activities of Qu-exosome-SPIONs/MF were evaluated both in vitro and in vivo. Results: The Qu-exosome-SPIONs were well constructed and harvested by magnetic separation with a uniform size and shape in a diameter of approximately 86.2 nm. The water solubility of quercetin increased 1.97-fold when loaded into the SPION-modified exosomes. The application of SPIONs/MF endowed the Qu-exosomes with favorable targeting capacity. In vitro studies showed that Qu-exosome-SPIONs/MF more effectively inhibited or attenuated ß cell apoptosis and promoted insulin secretion in response to elevated glucose (GLC) compared with quercetin or Qu-exosome-SPIONs. In vivo studies demonstrated that Qu-exosome-SPIONs/MF displayed an ideal pancreatic islet targeting capacity, thereby leading to the restoration of islet function. Conclusion: The Qu-exosome-SPIONs/MF nano-delivery system significantly enhanced the quercetin concentration in pancreatic islets and thereby improved pancreatic islet protection.

Hybrid cross-linked sodium carboxymethyl starch/polyacrylamide flexible sensing hydrogels with adhesion, antimicrobial properties and multiple responses.

Ionic hydrogels used as ideal and flexible strain sensor materials should have excellent mechanical, adhesive and antimicrobial properties. However, it is challenging to achieve these multifunctional requirements simultaneously. Herein, we designed and prepared a multifunctional ionic hydrogel with a multi-length tentacle bentonite backbone to initiate the free radical polymerization of acrylic acid bentonite (AABT) and acrylamide (AAm). The interactions of covalent cross-linking, hydrogen bonding cross-linking, charge interactions and physical entanglement between hybrid polyacrylamide-AABT (PAAm-AABT), sodium carboxymethyl starch (SCMS) and PAAm form an multi-in-one hybrid supramolecular network hydrogel (CABZ). This CABZ ion-conductive hydrogel is capable of detecting weak deformation with a detection limit of 1 % strain, high tensile properties of 995 %, excellent strength of 254.5 kPa, fast response (≈0.21 s), high sensitivity of 0.86 and high conductivity of 0.37 S/m. In addition, this CABZ ion-conductive hydrogel has impressive adhesion properties with shear adhesion strength up to 50.78 kPa and broad-spectrum antibacterial properties achieved by AABT-loaded ZnO nanoparticles. Through special AABT hybrid cross-linking, the CABZ ion-conductive hydrogel achieves stable mechanical properties, highly sensitive signal response and antimicrobial properties, which will make it a good choice for flexible wearable sensor materials.

Erratum: Cytochalasin H isolated from mangrove-derived endophytic fungus inhibits epithelial-mesenchymal transition and cancer stemness via YAP/TAZ signaling pathway in non-small cell lung cancer cells: Erratum.

[This corrects the article DOI: 10.7150/jca.50512.].

Multiparameter mobile blood analysis for complete blood count using contrast-enhanced defocusing imaging and machine vision.

Blood analysis through complete blood count is the most basic medical test for disease diagnosis. Conventional blood analysis requires bulky and expensive laboratory facilities and skilled technicians, limiting the universal medical use of blood analysis outside well-equipped laboratory environments. Here, we propose a multiparameter mobile blood analyzer combined with label-free contrast-enhanced defocusing imaging (CEDI) and machine vision for instant and on-site diagnostic applications. We designed a low-cost and high-resolution miniature microscope (size: 105 mm × 77 mm × 64 mm, weight: 314 g) that comprises a pair of miniature aspheric lenses and a 415 nm LED for blood image acquisition. The analyzer, adopting CEDI, can obtain both the refractive index distributions of the white blood cell (WBC) and hemoglobin spectrophotometric information, enabling the analyzer to supply rich blood parameters, including the five-part WBC differential count, red blood cell (RBC) count, and mean corpuscular hemoglobin (MCH) quantification with machine vision algorithms and the Lambert-Beer law. We have shown that our assay can analyze a blood sample within 10 minutes without complex staining, and measurements (30 samples) from the analyzer have a strong linear correlation with clinical reference values (significance level of 0.0001). This study provides a miniature, light weight, low-cost, and easy-to-use blood analysis technique that overcomes the challenge of simultaneously realizing FWD count, RBC count, and MCH analysis using a mobile device and has great potential for integrated surveillance of various epidemic diseases, including coronavirus infection, invermination, and anemia, especially in low- and middle-income countries.

Direction Selectivity of TmY Neurites in Drosophila.

The perception of motion is an important function of vision. Neural wiring diagrams for extracting directional information have been obtained by connectome reconstruction. Direction selectivity in Drosophila is thought to originate in T4/T5 neurons through integrating inputs with different temporal filtering properties. Through genetic screening based on synaptic distribution, we isolated a new type of TmY neuron, termed TmY-ds, that form reciprocal synaptic connections with T4/T5 neurons. Its neurites responded to grating motion along the four cardinal directions and showed a variety of direction selectivity. Intriguingly, its direction selectivity originated from temporal filtering neurons rather than T4/T5. Genetic silencing and activation experiments showed that TmY-ds neurons are functionally upstream of T4/T5. Our results suggest that direction selectivity is generated in a tripartite circuit formed among these three neurons-temporal filtering, TmY-ds, and T4/T5 neurons, in which TmY-ds plays a role in the enhancement of direction selectivity in T4/T5 neurons.

Label-free hematology analysis method based on defocusing phase-contrast imaging under illumination of 415†nm light.

Label-free imaging technology is a trending way to simplify and improve conventional hematology analysis by bypassing lengthy and laborious staining procedures. However, the existing methods do not well balance system complexity, data acquisition efficiency, and data analysis accuracy, which severely impedes their clinical translation. Here, we propose defocusing phase-contrast imaging under the illumination of 415 nm light to realize label-free hematology analysis. We have verified that the subcellular morphology of blood components can be visualized without complex staining due to the factor that defocusing can convert the second-order derivative distribution of samples' optical phase into intensity and the illumination of 415 nm light can significantly enhance the contrast. It is demonstrated that the defocusing phase-contrast images for the five leucocyte subtypes can be automatically discriminated by a trained deep-learning program with high accuracy (the mean F1 score: 0.986 and mean average precision: 0.980). Since this technique is based on a regular microscope, it simultaneously realizes low system complexity and high data acquisition efficiency with remarkable quantitative analysis ability. It supplies a label-free, reliable, easy-to-use, fast approach to simplifying and reforming the conventional way of hematology analysis.

Robust whole slide image analysis for cervical cancer screening using deep learning.

Computer-assisted diagnosis is key for scaling up cervical cancer screening. However, current recognition algorithms perform poorly on whole slide image (WSI) analysis, fail to generalize for diverse staining and imaging, and show sub-optimal clinical-level verification. Here, we develop a progressive lesion cell recognition method combining low- and high-resolution WSIs to recommend lesion cells and a recurrent neural network-based WSI classification model to evaluate the lesion degree of WSIs. We train and validate our WSI analysis system on 3,545 patient-wise WSIs with 79,911 annotations from multiple hospitals and several imaging instruments. On multi-center independent test sets of 1,170 patient-wise WSIs, we achieve 93.5% Specificity and 95.1% Sensitivity for classifying slides, comparing favourably to the average performance of three independent cytopathologists, and obtain 88.5% true positive rate for highlighting the top 10 lesion cells on 447 positive slides. After deployment, our system recognizes a one giga-pixel WSI in about 1.5 min.

Relevance of Physical Activities, Sedentary Behaviors, and Genetic Predisposition in Body Fatness: Population-Based Study on Chinese Adults.

INTRODUCTION: Little attention has been paid to the interacting effect of specific intensities of physical activities (PAs) and sedentary lifestyle, like television watching, and genetic predisposition on body composition indices among Chinese adults. Herein, we aimed to examine whether specific types of PAs and sedentary behaviors (SBs) were associated with body composition indices among Chinese adults and to further explore whether these associations interacted with the genetic predisposition to high BMI. METHODS: Cross-sectional data regarding PAs and time spent on SBs and dietary intake of 3,976 Chinese adults (54.9% women) aged 25-65 years in Southwest China were obtained via questionnaires in 2013-2015. Weight, height, and waist circumference (WC) were measured, and BMI, percentage of body fat (%BF), fat mass index (FMI), and fat-free mass index (FFMI) of the participants were calculated. Genetic risk score (GRS) was calculated on 9 established BMI-associated SNPs among Chinese adults. RESULTS: When the participants were stratified by GRS for BMI, significant associations were only found for adults with high GRS for BMI: moderate-to-vigorous physical activity (MVPA) was negatively associated with WC and %BF and positively related to FFMI. The adjusted positive relationship of time spent watching television with BMI, WC, %BF, and FMI were also just found between adults with high weighted GRS for high BMI: for every 1 h increment in television watching, the BMI, WC, %BF, and FMI of the participants increased by 0.2 kg/m2, 0.9 cm, 0.3%, and 0.1 kg/m2, respectively (p < 0.02). CONCLUSION: MVPA may be a protective factor against obesity, and prolonged television watching may accentuate adiposity. These putative effects may be more pronounced among individuals with a high genetic risk of a high BMI.

Glucocorticoid-induced dose-related and site-specific bone remodelling, microstructure, and mechanical changes in cancellous and cortical bones.

The study investigated the effects of long-term glucocorticoid (GC) administration on bone remodelling, microstructure, and biomechanical strength in cortical and cancellous (trabecular) bones. Thirty-one female Sprague-Dawley rats were randomly divided into three dexamethasone (Dex) dosage groups, 1.0, 2.5, and 5.0 mg/kg twice a week for 8 weeks, and one control group treated with saline. At the end of the experiment, the tibia of one side and the fourth lumbar vertebrae were processed into sections for a histomorphometric analysis, while the femur of the same side and the fifth vertebrae were isolated for a biomechanical test. A dose-dependent decline in bone formation was observed in both trabecular and cortical (periosteal and endosteal) bones. In contrast, bone resorption was inhibited only in cancellous bone in the two higher dose groups and not dose-related. The ratio of Node/Termini increased, while marrow star volume (MSV) decreased in all Dex groups in metaphyseal trabecular bones, both of which were dose-dependent. Subendosteal cortex porosity increased in parallel with non-uniform trabecular distribution, but cortical thickness remained unchanged. Interestingly, there were no significant changes in microstructure or mechanical strength in lumbar trabecular bone. The cortical elastic load was dose-independently reduced in all three Dex groups when compared with the control group. In summary, bone remodelling was dose-dependently inhibited in cancellous bones but enhanced in intracortical bones. The non-uniform distribution of trabecular bone and increased porosity in the inner edge of cortical bone were both in parallel with GC dosage, and the porosity increase was more likely to occur, leading to reduced cortical mechanical strength.

Predicting and Exploring the Mechanisms of Erzhi Pill in Prevention and Treatment of Osteoporosis Based on Network Pharmacology and Zebrafish Experiments.

BACKGROUND: Erzhi Pill (EZP), a traditional Chinese medicine (TCM) prescription, has been widely applied to improve bone metabolism and treat osteoporosis (OP) in China. However, its effective constituents and mechanisms remain unclear. METHODS: By combining network pharmacology and zebrafish experiments, an integrative method was employed to address this problem. Firstly, the disease targets of OP were collected from two public gene databases. Secondly, the active compounds and drug targets of EZP were obtained from the traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP). Thirdly, a drug-target-disease interaction network was constructed, and the key active components were identified by analyzing the topological characteristics of the network. Finally, these predicted results were tested by zebrafish experiments and compared with those from the literature. Specifically, quercetin as an important representative active component of EZP was applied to wild type and transgenic zebrafish larvae to assess its effects on skull mineralization and osteoplastic differentiation. RESULTS: Our study identified 72 active compounds, 220 targets and 166 signaling pathways probably involved in the prevention and treatment of OP by EZP, wherein quercetin, apigenin, daidzein, luteolin, ursolic acid and kaempferol could be the key compounds, while PI3K-Akt signaling pathway, TNF signaling pathway and IL-17 signaling pathway could be the key signaling pathways. The experiments indicated that quercetin attenuated both the decrease of skull mineralization and the inhibition of skull osteoplastic differentiation in zebrafish larvae trigged by dexamethasone. CONCLUSION: Our study not only investigated potentially effective constituents and mechanisms of EZP in the prevention and treatment of OP, but also provided a reference for the in-depth research, development and application of TCM.

Chemical sectioning fluorescence tomography: high-throughput, high-contrast, multicolor, whole-brain imaging at subcellular resolution.

A thorough neuroanatomical study of the brain architecture is crucial for understanding its cellular compositions, connections, and working mechanisms. However, the fine- and multiscale features of neuron structures make it challenging for microscopic imaging, as it requires high contrast and high throughput simultaneously. Here, we propose chemical sectioning fluorescence tomography (CSFT) to solve this problem. By chemically switching OFF/ON the fluorescent state of the labeled proteins (FPs), we light only the top layer as thin as submicron for imaging without background interference. Combined with the wide-field fluorescence micro-optical sectioning tomography (fMOST) system, we have shown multicolor CSFT imaging. We also demonstrate mouse whole-brain imaging at the subcellular resolution, as well as the power for quantitative acquisition of synaptic-connection-related pyramidal dendritic spines and axon boutons on the brain-wide scale at the complete single-neuron level. We believe that the CSFT method would greatly facilitate our understanding of the brain-wide neuron networks.

High throughput slanted scanning whole slide imaging system for digital pathology.

In whole slide imaging (WSI), normally only a one layer imaging of the slide is performed. Autofocus at multiple positions is usually required. But defocus blur still exists due to tissue folding or specimen thickness. Repeated Z-stack scan be applied here, which, however, is too time consuming. Here, a high throughput slanted scanning WSI system is reported. In this system, the slide surface was slanted 1° relative to the focal plane. Thus, the focal plane spanned multiple layers of the sample. By moving the slide, multi-layer image data of the sample can be acquired simultaneously at a time frame comparable to conventional 1-layer imaging. With image fusion, defocus blur can be avoided. High quality and fast imaging of both cytological and histological slide specimens was demonstrated without applying aberration correction. The system can be a highly efficient way for the application of WSI in digital pathology.

Cytochalasin H isolated from mangrove-derived endophytic fungus inhibits epithelial-mesenchymal transition and cancer stemness via YAP/TAZ signaling pathway in non-small cell lung cancer cells.

Our previous studies have isolated cytochalasin H (CyH) from endophytic fungus derived from mangrove and found that CyH induced apoptosis and inhibited migration and angiogenesis in non-small cell lung cancer (NSCLC) cells. In this study, we further investigated the effect of CyH on epithelial-mesenchymal transition (EMT) and cancer stemness of A549 and NCI-H460 NSCLC cells and the underlying mechanisms, especially the role of YAP/ TAZ signaling pathway in the process. Our results showed that CyH significantly inhibited invasive ability and the sphere formation of NSCLC cells. The expression of E-cadherin, an EMT epithelial marker, was obviously up-regulated, while the expression of Vimentin and N-cadherin, the EMT mesenchymal markers, was dramatically down-regulated by CyH treatment in NSCLC cells. Moreover, the expression of EMT-associated transcription factors including Slug, Twist1, and Snail1 and stemness markers including Nanog, Sox-2, and Oct-4 was significantly down-regulated by CyH treatment in NSCLC cells. Additionally, CyH significantly down-regulated YAP and TAZ expression and up-regulated LAST1/2 and MST1/2 expression, and CyH inhibited the interaction between YAP and TEAD. Furthermore, YAP knockdown abolished the effect of CyH on the expression of EMT- and stemness-related markers in NSCLC cells. Taken together, these results suggest that CyH inhibits EMT and cancer stemness of NSCLC cells via the regulation of YAP/TAZ signaling pathway.

Divergent Projection Patterns Revealed by Reconstruction of Individual Neurons in Orbitofrontal Cortex.

The orbitofrontal cortex (OFC) is involved in diverse brain functions via its extensive projections to multiple target regions. There is a growing understanding of the overall outputs of the OFC at the population level, but reports of the projection patterns of individual OFC neurons across different cortical layers remain rare. Here, by combining neuronal sparse and bright labeling with a whole-brain florescence imaging system (fMOST), we obtained an uninterrupted three-dimensional whole-brain dataset and achieved the full morphological reconstruction of 25 OFC pyramidal neurons. We compared the whole-brain projection targets of these individual OFC neurons in different cortical layers as well as in the same cortical layer. We found cortical layer-dependent projections characterized by divergent patterns for information delivery. Our study not only provides a structural basis for understanding the principles of laminar organizations in the OFC, but also provides clues for future functional and behavioral studies on OFC pyramidal neurons.

GTree: an Open-source Tool for Dense Reconstruction of Brain-wide Neuronal Population.

Recent technological advancements have facilitated the imaging of specific neuronal populations at the single-axon level across the mouse brain. However, the digital reconstruction of neurons from a large dataset requires months of manual effort using the currently available software. In this study, we develop an open-source software called GTree (global tree reconstruction system) to overcome the above-mentioned problem. GTree offers an error-screening system for the fast localization of submicron errors in densely packed neurites and along with long projections across the whole brain, thus achieving reconstruction close to the ground truth. Moreover, GTree integrates a series of our previous algorithms to significantly reduce manual interference and achieve high-level automation. When applied to an entire mouse brain dataset, GTree is shown to be five times faster than widely used commercial software. Finally, using GTree, we demonstrate the reconstruction of 35 long-projection neurons around one injection site of a mouse brain. GTree is also applicable to large datasets (10 TB or higher) from various light microscopes.

The Nutrition and Health in Southwest China (NHSC) study: design, implementation, and major findings.

BACKGROUND: There are few studies of nutritional and genetic factors and their interactions on the risk of noncommunicable diseases (NCDs) among Chinese adults. OBJECTIVE: Our aim for the Nutrition and Health in Southwest China (NHSC) study is to investigate the impact of diet, lifestyle, genetic background, and their interactions on NCDs among adults in Southwest China. METHODS: The NSHC is a prospective cohort study initiated in winter 2013. The baseline data collection was completed in December 2018, and follow-ups are conducted every 2 years. Information on genomics, anthropometry, nutrition, eating behaviors, physical activity, depression and mental disorders, clinical and biochemical examinations, and lifestyles was collected. RESULTS: 7926 adults completed the baseline questionnaire. The average age of participants was 42.6 (9.8) years at study enrollment. More than half were female, 37.2% had achieved more than 12 years of education, and 49.3% of them came from family income >35,000 Yuan. Our analyses of the baseline data suggested that adults with greater dietary energy density appeared to have greater body mass index, fat mass index, fat-free mass index and percentage body fat, and that participants with a higher level of dietary glycemic index, glycemic load, or serum 25(OH)D had a less favorable glucose homeostasis. In addition, spending less time watching television and having a healthy eating pattern may play significant roles in later cellular aging. CONCLUSIONS: In conclusion, the NHSC cohort provides valuable data for investigations of the relevance of gene, nutrition, lifestyles, and their interactions on NCDs among southwestern Chinese adults.

Extending the 3D scanning range of DMD-based scanners for femtosecond lasers.

Digital micromirror devices (DMDs) have shown their potential in 2-photon imaging and microfabrication as diffractive scanners for femtosecond lasers. However, the scanning range of a DMD-based scanner is decreased by the spatial filter (SF) used to block undesired diffraction orders. Instead of an SF, we present a method of introducing and correcting aberration (ICA) to reduce the effects of these undesired diffraction orders. In ICA, aberrations are introduced by optical elements, and only the aberration of the desired diffraction order is corrected by adding a compensatory phase to the scanning phase. The scanning ranges in the y and z directions can be nearly doubled when the SF is removed. We demonstrate that ICA can be conveniently applied to a previously constructed DMD-based 2-photon microscope, and the field of view can be extended at different axial positions.

Aberration-corrected three-dimensional non-inertial scanning for femtosecond lasers.

Large aberrations are induced by non-collimated light when the convergence or divergence of the incident beam on the back-pupil plane of the objective lens is adjusted for 3D non-inertial scanning. These aberrations significantly degrade the focus quality and decrease the peak intensity of the femtosecond laser focal spot. Here, we describe an aberration-corrected 3D non-inertial scanning method for femtosecond lasers based on a digital micromirror device (DMD) that is used for both beam scanning and aberration correction. An imaging setup is used to detect the focal spot in the 3D space, and an iterative optimization algorithm is used to optimize the focal spot. We demonstrate the application of our proposed approach in two-photon imaging. With correction for the 200-µm out-of-focal plane, the optical axial resolution improves from 7.67 to 3.25 µm, and the intensity of the fluorescence signal exhibits an almost fivefold improvement when a 40× objective lens is used. This aberration-corrected 3D non-inertial scanning method for femtosecond lasers offers a new approach for a variety of potential applications, including nonlinear optical imaging, microfabrication, and optical storage.