Exosomes Derived from Astragaloside IV-pretreated Endothelial Progenitor Cells (AS-IV-Exos) Alleviated Endothelial Oxidative Stress and Dysfunction Via the miR-210/ Nox2/ROS Pathway.

BACKGROUND: Chronic hyperglycemia in diabetes induces oxidative stress, leading to damage to the vascular system. In this study, we aimed to evaluate the effects and mechanisms of AS-IV-Exos in alleviating endothelial oxidative stress and dysfunction caused by high glucose (HG). METHODS: Histopathological changes were observed using HE staining, and CD31 expression was assessed through immunohistochemistry (IHC). Cell proliferation was evaluated through CCK8 and EDU assays. The levels of ROS, SOD, and GSH-Px in the skin tissues of each group were measured using ELISA. Cell adhesion, migration, and tube formation abilities were assessed using adhesion, Transwell, and tube formation experiments. ROS levels in HUVEC cells were measured using flow cytometry. The levels of miR-210 and Nox2 were determined through quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The expression of Nox2, SOD, GSH-Px, CD63, and CD81 was confirmed using WB. RESULTS: The level of miR-210 was reduced in diabetes-induced skin damage, while the levels of Nox2 and ROS increased. Treatment with AS-IV increased the level of miR-210 in EPC-Exos. Compared to Exos, AS-IV-Exos significantly reduced the proliferation rate, adhesion number, migration speed, and tube-forming ability of HGdamaged HUVEC cells. AS-IV-Exos also significantly decreased the levels of SOD and GSH-Px in HG-treated HUVEC cells and reduced the levels of Nox2 and GSH-Px. However, ROS levels and Nox2 could reverse this effect. CONCLUSION: AS-IV-Exos effectively alleviated endothelial oxidative stress and dysfunction induced by HG through the miR-210/Nox2/ROS pathway.

Effect and Mechanism of Sodium Butyrate on Neuronal Recovery and Prognosis in Diabetic Stroke.

Ischemic stroke is a cerebrovascular lesion caused by local ischemia and hypoxia. Diabetes mellitus (DM) is a chronic inflammatory disease that disturbs immune homeostasis and predisposes patients to ischemic stroke. The mechanism by which DM exacerbates stroke remains unclear, although it may involve disturbances in immune homeostasis. Regulatory T cells (Tregs) play a regulatory role in many diseases, but the mechanism of Tregs in diabetes complicated by stroke remains unclear. Sodium butyrate is a short-chain fatty acid that increases Treg levels. This study examined the role of sodium butyrate in the prognosis of neurological function in diabetic stroke and the mechanism by which Tregs are amplified in the bilateral cerebral hemispheres. We evaluated the brain infarct volume, observed 48-h neuronal injury and 28-day behavioral changes, and calculated the 28-day survival rate in mice. We also measured Treg levels in peripheral blood and brain tissue, recorded changes in the blood‒brain barrier and water channel proteins and neurotrophic changes in mice, measured cytokine levels and peripheral B-cell distribution in bilateral hemispheres and peripheral blood, and examined the polarization of microglia and the distribution of peripheral T-cell subpopulations in bilateral hemispheres. Diabetes significantly exacerbated the poor prognosis and neurological deficits in mice with stroke, and sodium butyrate significantly improved infarct volume, prognosis, and neurological function and showed different mechanisms in brain tissue and peripheral blood. The potential regulatory mechanism in brain tissue involved modulating Tregs/TGF-ß/microglia to suppress neuroinflammation, while that in peripheral blood involved improving the systemic inflammatory response through Tregs/TGF-ß/T cells.

Stellate ganglion block reduces inflammation and improves neurological function in diabetic rats during ischemic stroke.

Diabetes mellitus is an independent risk factor for ischemic stroke. Both diabetes mellitus and stroke are linked to systemic inflammation that aggravates patient outcomes. Stellate ganglion block can effectively regulate the inflammatory response. Therefore, it is hypothesized that stellate ganglion block could be a potential therapy for ischemic stroke in diabetic subjects. In this study, we induced diabetes mellitus in rats by feeding them a high-fat diet for 4 successive weeks. The left middle cerebral artery was occluded to establish models of ischemic stroke in diabetic rats. Subsequently, we performed left stellate ganglion block with 1% lidocaine using the percutaneous posterior approach 15 minutes before reperfusion and again 20 and 44 hours after reperfusion. Our results showed that stellate ganglion block did not decrease the blood glucose level in diabetic rats with diabetes mellitus but did reduce the cerebral infarct volume and the cerebral water content. It also improved the recovery of neurological function, increased 28-day survival rate, inhibited Toll like receptor 4/nuclear factor kappa B signaling pathway and reduced inflammatory response in the plasma of rats. However, injection of Toll like receptor 4 agonist lipopolysaccharide 5 minutes before stellate ganglion block inhibited the effect of stellate ganglion block, whereas injection of Toll like receptor 4 inhibitor TAK242 had no such effect. We also found that stellate ganglion block performed at night had no positive effect on diabetic ischemic stroke. These findings suggest that stellate ganglion block is a potential therapy for diabetic ischemic stroke and that it may be mediated through the Toll like receptor 4/nuclear factor kappa B signaling pathway. We also found that the therapeutic effect of stellate ganglion block is affected by circadian rhythm.

Dual Recognition DNA Triangular Prism Nanoprobe: Toward the Relationship between K+ and pH in Lysosomes.

Lysosomal acidification is essential for its degradative function, and the flux of H+ correlated with that of K+ in lysosomes. However, there is little research on their correlation due to the lack of probes that can simultaneously image these two ions. To deeply understand the role of K+ in lysosomal acidification, here, we designed and fabricated a nanodevice using a K+-aptamer and two pH-triggered nanoswitches incorporated into a DNA triangular prism (DTP) as a dual signal response platform to simultaneously visualize K+ and pH in lysosomes by a fluorescence method. This strategy could conveniently integrate two signal recognition modules into one probe, so as to achieve the goal of sensitive detection of two kinds of signals in the same time and space, which is suitable for the detection of various signals with the correlation of concentration. By co-imaging both K+ and H+ in lysosomes, we found that the efflux of K+ was accompanied by a decrease of pH, which is of great value in understanding lysosomal acidification. Moreover, this strategy also has broad prospects as a versatile optical sensing platform for multiplexed analysis of other biomolecules in living cells.

Characterization of HPV subtypes not covered by the nine-valent vaccine in patients with CIN 2-3 and cervical squamous cell carcinoma.

BACKGROUND: As the second most common female malignant tumor, cervical cancer is also one of the most preventable and avoidable cancers. The World Health Organization has launched a global plan to accelerate the elimination of cervical cancer. Therefore, in the era of postvaccine, the role of HPV subtypes in cervical precancerous lesions and cervical cancer that are not covered by vaccine should be further discussed. The purpose of this study was to explore the role of HPV subtypes not covered by the nine-valent vaccine in high-grade cervical precancerous lesions and cervical cancer. MATERIALS AND METHODS: A retrospective analysis was performed on the clinical data of 5220 patients with an HPV infection who were diagnosed and treated in the Department of Gynecology of Shanghai General Hospital between October 2016 and February 2020. In addition, the clinical characteristics of the biopsy results of 470 cases of cervical intraepithelial neoplasia (CIN) 2-3 and 205 cases of cervical squamous cell carcinoma were analyzed. RESULTS: Among patients with HPV subtype infection not covered by the nine-valent vaccine, univariate analysis showed that compared with patients with CIN 2-3, age ≥ 50, not using condom and TCT reported as ASC-H were risk factors for cervical squamous cell carcinoma (P < 0.05). The detection rates of HPV subtype not covered by the nine-valent vaccine in CIN 2-3 and cervical squamous cell carcinoma patients were 7.23% and 6.34%, respectively. CONCLUSION: In patients with CIN 2-3 and cervical squamous cell carcinoma, the infection rates of HPV subtype not covered by the nine-valent vaccine were 7.23% and 6.34%, respectively. With the increasing popularity of the vaccine, the infection rates of the corresponding HPV subtype decreased; however, HPV subtype infection not covered by the nine-valent vaccine should not be ignored.

Target-Dependent Gating of Nanopores Integrated with H-Cell: Toward A General Platform for Photoelectrochemical Bioanalysis.

Herein we present a proof-of-concept study of target-dependent gating of nanopores for general photoelectrochemical (PEC) bioanalysis in an H-cell. The model system was constructed upon a left chamber containing ascorbic acid (AA), the antibody modified porous anodic alumina (AAO) membrane separator, and a right chamber placed with the three-electrode system. The sandwich immunocomplexation and the associated enzymatic generation of biocatalytic precipitation (BCP) in the AAO nanopores would regulate the diffusion of AA from the left cell to the right cell, leading to a varied photocurrent response of the ZnInS nanoflakes photoelectrode. Exemplified by fatty-acid-banding protein (FABP) as the target, the as-developed protocol achieved good performance in terms of sensitivity, selectivity, reproducibility, as well as efficient reutilization of the working electrode. On the basis of an H-cell, this work features a new protocol of target-dependent gating-based PEC bioanalysis, which can serve as a general PEC analytical platform for various other targets of interest.

Establishing Interfacial Charge-Transfer Transitions on Ferroelectric Perovskites: An Efficient Route for Photoelectrochemical Bioanalysis.

This work presents the concept of establishing interfacial charge-transfer transitions (ICTT) on ferroelectric perovskites for efficient photoelectrochemical (PEC) bioanalysis. The model system was exemplified by using representative lead titanate (PbTiO3) and an enzyme tandem consisting of the isocitrate dehydrogenase (ICDH) and p-hydroxybenzoate hydroxylase (PHBH). The enzymatic generation of protocatechuic acid (PCA) can coordinate onto the surface of the PbTiO3 and hence form the ICTT that enables direct ligand-to-metal charge transfer from the highest occupied molecular orbital (HOMO) of PCA to the conduction band (CB) of PbTiO3 under light irradiation. Due to the ferroelectric polarization induced electric field of PbTiO3 and the surface polarity of PCA modification, enhanced charge separation of the ICTT contributes to the generation of anodic photocurrent and thus underlies a unique route for detecting the enzymatic activity or its substrate. For dehydrogenase detection, this strategy has better performance than some classical methodologies in terms of high sensitivity and improved selectivity. This work not only features ICTT establishment on ferroelectric perovskites for unique bioanalysis but also provides new insights into the utilization of ferroelectric perovskites for advanced PEC bioanalysis.

Three-dimensional CdS nanosheet-enwrapped carbon fiber framework: Towards split-type CuO-mediated photoelectrochemical immunoassay.

This work reports a customized methodology for the fabrication of 3D CdS nanosheet (NS)-enwrapped carbon fiber framework (CFF) and its utilization for sensitive split-type CuO-mediated PEC immunoassay. Specifically, the 3D CdS NS-CFF was fabricated via a solvothermal process, while the sandwich immunocomplexing was allowed in a 96 well plate with CuO nanoparticles (NPs) as the signaling labels. The subsequent release of the Cu2+ ions was directed to interact with the CdS NS, generating trapping sites and thus inhibiting its photocurrent generation. In such a protocol, the 3D CdS NS-CFF photoelectrode could not only guarantee its sufficient contact with the Cu2+-containing solution but also supply plenty CdS surface for the Cu2+ ions. Because of the target-dependent release of the Cu2+ ions and its proper coupling with the 3D CdS NS-CFF photoelectrode, a sensitive split-type PEC immunoassay was achieved for the detection of brain natriuretic peptide (BNP). This proposed system exhibited good stability and selectivity, and its applicability for real sample analysis was also demonstrated via comparison with the commercial BNP enzyme-linked immunosorbent assay (ELISA) kit. We expect this work could stimulate more interest in the design and utilization of 3D photoelectrodes for novel PEC bioanalysis.

Salvianolic acid B improves the disruption of high glucose-mediated brain microvascular endothelial cells via the ROS/HIF-1α/VEGF and miR-200b/VEGF signaling pathways.

The study investigated the roles and mechanisms of Salvianolic acid B (Sal B) on permeability of rat brain microvascular endothelial cells (RBMECs) exposed to high glucose. The results demonstrated that Sal B greatly up-regulated the expression of tight junction (TJ) proteins and decreased the permeability of RBMECs compared with the control group. And the increase of reactive oxidative species (ROS) production, the upregulation of hypoxia-inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) protein induced by high glucose were antagonized by Sal B. In addition, a great decrease of microRNA-200b (miR-200b) was observed in the RBMECs under high-glucose condition, which was significantly increased by Sal B pretreatment. And overexpression of miR-200b markedly attenuated the RBMECs permeability and inhibited the expression of VEGF protein by targeting with 3'-UTR of its mRNA. This led to the conclusion that Sal B-mediated improvement of blood-brain barrier dysfunction induced by high-glucose is related to the ROS/HIF-1α/VEGF and miR-200b/VEGF signaling pathways.

Enhancing the bioactivity of Poly(lactic-co-glycolic acid) scaffold with a nano-hydroxyapatite coating for the treatment of segmental bone defect in a rabbit model.

PURPOSE: Poly(lactic-co-glycolic acid) (PLGA) is excellent as a scaffolding matrix due to feasibility of processing and tunable biodegradability, yet the virgin scaffolds lack osteoconduction and osteoinduction. In this study, nano-hydroxyapatite (nHA) was coated on the interior surfaces of PLGA scaffolds in order to facilitate in vivo bone defect restoration using biomimetic ceramics while keeping the polyester skeleton of the scaffolds. METHODS: PLGA porous scaffolds were prepared and surface modification was carried out by incubation in modified simulated body fluids. The nHA coated PLGA scaffolds were compared to the virgin PLGA scaffolds both in vitro and in vivo. Viability and proliferation rate of bone marrow stromal cells of rabbits were examined. The constructs of scaffolds and autogenous bone marrow stromal cells were implanted into the segmental bone defect in the rabbit model, and the bone regeneration effects were observed. RESULTS: In contrast to the relative smooth pore surface of the virgin PLGA scaffold, a biomimetic hierarchical nanostructure was found on the surface of the interior pores of the nHA coated PLGA scaffolds by scanning electron microscopy. Both the viability and proliferation rate of the cells seeded in nHA coated PLGA scaffolds were higher than those in PLGA scaffolds. For bone defect repairing, the radius defects had, after 12 weeks implantation of nHA coated PLGA scaffolds, completely recuperated with significantly better bone formation than in the group of virgin PLGA scaffolds, as shown by X-ray, Micro-computerized tomography and histological examinations. CONCLUSION: nHA coating on the interior pore surfaces can significantly improve the bioactivity of PLGA porous scaffolds.

[Research on relevance between macroscopic feature individual differences and quality of Gansu cultivated Angelica sinensis].

OBJECTIVE: To analyze the appearance differences of Gansu cultivated Angelica sinensis, and explore the relevance between the appearance differences and quality. METHODS: The macroscopic feature of 22 batches of Angelica sinensis from different habitats was measured as index. The content of ferulic acid, volatile oil and extract were determined by the method recorded in the Chinese Pharmacopoeia. The data were analyzed by SPSS 15.0 software. RESULTS: The habitat was positively correlated with the index. Indexes of nine groups had direct correlation with each other. The habitat was significantly correlated with other indexes except the length of the head. The extract and ferulic acid were positively correlated with habitat and index. Extract had significant correlation with macroscopic feature. Ferulic acid only had significant correlation with head length. The volatile oil only had significant correlation with habitat and no significant correlation with index. Root weight and number of lateral roots had obvious difference in different habitat which coefficient of variation was 44.1% and 28.6%, respectively. CONCLUSION: There are significant individual differences in Angelica sinensis. The chemical composition has a certain correlation with macroscopic feature. Angelica sinensis cultivation needs to consider the choice of habitat.

[Simulation of vegetation indices optimizing under retrieval of vegetation biochemical parameters based on PROSPECT + SAIL model].

This study analyzed the sensitivities of three vegetation biochemical parameters [chlorophyll content (Cab), leaf water content (Cw), and leaf area index (LAI)] to the changes of canopy reflectance, with the effects of each parameter on the wavelength regions of canopy reflectance considered, and selected three vegetation indices as the optimization comparison targets of cost function. Then, the Cab, Cw, and LAI were estimated, based on the particle swarm optimization algorithm and PROSPECT + SAIL model. The results showed that retrieval efficiency with vegetation indices as the optimization comparison targets of cost function was better than that with all spectral reflectance. The correlation coefficients (R2) between the measured and estimated values of Cab, Cw, and LAI were 90.8%, 95.7%, and 99.7%, and the root mean square errors of Cab, Cw, and LAI were 4.73 microg x cm(-2), 0.001 g x cm(-2), and 0.08, respectively. It was suggested that to adopt vegetation indices as the optimization comparison targets of cost function could effectively improve the efficiency and precision of the retrieval of biochemical parameters based on PROSPECT + SAIL model.

[Estimation of leaf area index by normalized composite vegetation index fusing the spectral feature of canopy water content].

The accurate inversion of leaf area index (LADI) in canopy is very important for guiding crop management and assessing crop yield. Sixty samples belonging to corn in four different areas of Jilin City were scanned by ASD field pro3 and LAI-2000 for optical data and LAI. A new vegetation index, the normalized composite Vegetation index (NCVI), containing the factor of canopy water content, is proposed in the present paper for a better quantitative estimation of LAI than with the remotely sensed normalized difference vegetation index (NDVI), especially in the arid and semi-arid areas. A model was built for inversion of LAI with NCVI, and experience validation. The results showed that there was a good linear correlation between the simulation LAI inversed from NCVI model and the real LAI values. The model breaking the limitations of the traditional empirical models for LAI inversion has a good result for estimating LAI of the dense canopy whose LAI value was greater than 3. In addition, NCVI model was very sensitive to the water environment of soil, and the inversion result in the arid and semi-arid areas was superior to the general area.

[Analysis and modeling of hyperspectral singularity in rice under Cd pollution].

In order to detect the Cd stress levels of rice growing in natural environment fast and accurately, based on wavelet transform technology in the visible light and near-infrared region (NIR), a method of identifying stress levels of rice under Cd pollution was explored. The hyperspectral data, biochemical parameters and heavy metals concentration in folium were collected for the rice growing in natural farmlands. Wavelet transform of hyperspectral reflectance (350-1 300 nm) was performed by using Daubechies 5 mother function and wavelet energy coefficients of spectral reflectance were extracted. In addition, the model between wavelet energy coefficient and Cd content was established. The result showed that the wavelet coefficients of the fifth decomposition level (d5) proved successful for detecting Cd pollution of rice; the singularity range of rice located in the region around 550-810 nm of spectral signal under Cd pollution; and the singularity amplitude was 0.04; The centre of modulus maxima located at 700 nm. Regression model based on third level wavelet energy coefficient can estimate the Cd content of rice accurately with the coefficient of determination (R2) of 0.958, and root mean square error (RMSE) of 0.122. It can be concluded that the singularity analysis technology applying wavelet transform to reflectance has been shown to be very promising in detecting rice under Cd pollution effectively, and wavelet energy coefficients can estimate Cd content of rice, and provide important reference for detecting other metal-induced stress on crop.

[Multi-diagnosis space models of As stress in rice based on hyperspectral indices].

High arsenic content in rice can influence the chlorophyll, water content and structure in their leaves, reduce the rate of photosynthesis and change their spectral features. Multiple models for diagnosing As contamination in rice based on spectral parameters were studied. Sixty samples belonging to mature rice in three different areas were scanned by ASD field pro3 for optical data. Arsenic reference values were obtained by atomic absorption spectrometry. First, correlation analysis was used between 9 hyperspectral indices and As content in rice, and three indices (PSNDa, fWBI, SIPI) were extracted to diagnose As contamination in rice, which were respectively sensitive to chlorophyll, water content and structure of leaves, then took the three indices to form a diagnosis spectral indices space (PSNDa-fWBI, PSNDa-SIPI, fWBI-SIPI) of As stress in rice. Second, principal component analysis and independent component analysis were also applied in these 9 hyperspectral indices, and two principal components (F1, F2) and two independent components(ICA1, ICA2) were extracted. These four components (F1, F2, ICA1, ICA2) were all correlated with As content in rice, and composed another two diagnosis spaces (F1-F2, ICA1-ICA2) for predicting As contamination. And these spaces composed a multiple diagnosis space model which diagnosed As contamination in rice of test area from different level, and showed a good result.

[Research on hyperspectral information parameters of chlorophyll content of rice leaf in Cd-polluted soil environment].

The remote sensing pollution mechanism in Cd-polluted soil is discussed depending on the research into the chlorophyll content of Cd-polluted rice leaf in the present paper. The response models of remote sensing information parameters, which reflected chlorophyll content variety of rice canopy with soil Cd pollution degree, were established based on Hyperion satellite data and a great number of ground experiment data. To extract sensitive remote sensing parameters for Cd pollution, multiple discriminant analysis (MDA) was applied to the reflectivity of 447-925 nm in Hyperion data and five remote sensing information parameters, including MCARI, NPCI, RVSI, NDVI and Depth671. Experiments indicated that MCARI is the most sensitive parameter to the chlorophyll content of Cd-polluted rice, whose response coefficient is 0.59. In the extent of 1.0-2.0 mg x kg(-1) of Cd pollution concentration in soil, MCARI curve shows a small decline. In the extent of 2.0-3.0 mg x kg(-1) of Cd pollution concentration in soil, MCARI curve is horizontal. Above 3.0 mg x kg(-1), MCARI shows a significant drop trend and so on. The research results showed that the chlorophyll content is a good indicator for nutrition situation of plant, capacity of photosynthesis and each developmental stage. And the chlorophyll remote sensing parameters in crop have a great significance for monitoring heavy metal pollution This study will help improve the precision and limitation of statistical methods and provide theoretical basis for and technical approach to monitoring soil Cd pollution in large area using hyperspectral remote sensing technology. However, the precision of pollution model needs to be improved.