Rapid Surface Charge Mapping Based on a Liquid Crystal Microchip.

Rapid surface charge mapping of a solid surface remains a challenge. In this study, we present a novel microchip based on liquid crystals for assessing the surface charge distribution of a planar or soft surface. This chip enables rapid measurements of the local surface charge distribution of a charged surface. The chip consists of a micropillar array fabricated on a transparent indium tin oxide substrate, while the liquid crystal is used to fill in the gaps between the micropillar structures. When an object is placed on top of the chip, the local surface charge (or zeta potential) influences the orientation of the liquid crystal molecules, resulting in changes in the magnitude of transmitted light. By measuring the intensity of the transmitted light, the distribution of the surface charge can be accurately quantified. We calibrated the chip in a three-electrode configuration and demonstrated the validity of the chip for rapid surface charge mapping using a borosilicate glass slide. This chip offers noninvasive, rapid mapping of surface charges on charged surfaces, with no need for physical or chemical modifications, and has broad potential applications in biomedical research and advanced material design.

Effects of Land Use Type Transformation on the Structure and Diversity of Soil Bacterial Communities.

Soil microbiota are significantly influenced by their microenvironments. Therefore, to understand the impacts of various land use patterns on the diversity and composition of soil bacterial communities, this study focused on three typical land use types-NF (natural forest), AF (artificial forests), and FL (farmland)-in the Heilongjiang Central Station Black-billed Capercaillie National Nature Reserve, located in the southwestern part of Heihe City, Heilongjiang Province, China. Using high-throughput sequencing of the 16S rRNA gene, we examined the soil bacterial community structures in these different land use types and explored their correlation with soil environmental factors. The following were our main observations: (1) Significant variations in soil chemical properties among different land use patterns were observed. In artificial forests, total nitrogen (TN), alkali hydrolyzed nitrogen (AN), total phosphorus (TP), and available phosphorus (AP) were higher compared to farmland and significantly higher than those in natural forests. Furthermore, the organic carbon content (SOC) in natural forests was higher than in artificial forests and significantly higher than in farmland. (2) Comparative analysis using the Shannon and Simpson indices revealed that bacterial community diversity was higher in artificial forests than in natural forests, which was significantly higher than in farmland. (3) The effect of different land use types on soil bacterial community structure was not significant. The three land types were dominated by Proteobacteria, Acidobacteria, and Actinobacteria. Proteobacteria exhibited a higher relative abundance in farmland and artificial forests compared to natural forests, whereas Actinobacteria exhibited the lowest relative abundance in natural forests. (4) Redundancy analysis (RDA) revealed that SOC, TN, AN, and AP were key environmental factors influencing the microbial communities of soil. Collectively, our findings demonstrated that land use practices can significantly alter soil nutrient levels, thereby influencing the structure of bacterial communities.

BpGRP1 acts downstream of BpmiR396c/BpGRF3 to confer salt tolerance in Betula platyphylla.

Glycine-rich RNA-binding proteins (GRPs) have been implicated in the responses of plants to environmental stresses, but the function of GRP genes involved in salt stress and the underlying mechanism remain unclear. In this study, we identified BpGRP1 (glycine-rich RNA-binding protein), a Betula platyphylla gene that is induced under salt stress. The physiological and molecular responses to salt tolerance were investigated in both BpGRP1-overexpressing and suppressed conditions. BpGRF3 (growth-regulating factor 3) was identified as a regulatory factor upstream of BpGRP1. We demonstrated that overexpression of BpGRF3 significantly increased the salt tolerance of birch, whereas the grf3-1 mutant exhibited the opposite effect. Further analysis revealed that BpGRF3 and its interaction partner, BpSHMT, function upstream of BpGRP1. We demonstrated that BpmiR396c, as an upstream regulator of BpGRF3, could negatively regulate salt tolerance in birch. Furthermore, we uncovered evidence showing that the BpmiR396c/BpGRF3 regulatory module functions in mediating the salt response by regulating the associated physiological pathways. Our results indicate that BpmiR396c regulates the expression of BpGRF3, which plays a role in salt tolerance by targeting BpGRP1.

Bp-miR408a participates in osmotic and salt stress responses by regulating BpBCP1 in Betula platyphylla.

The microRNAs, which are small RNAs of 18-25 nt in length, act as key regulatory factors in posttranscriptional gene expression during plant growth and development. However, little is known about their regulatory roles in response to stressful environments in birch (Betula platyphylla). Here, we characterized and further explored miRNAs from osmotic- and salt-stressed birch. Our analysis revealed a total of 190 microRNA (miRNA) sequences, which were classified into 180 conserved miRNAs and 10 predicted novel miRNAs based on sequence homology. Furthermore, we identified Bp-miR408a under osmotic and salt stress and elucidated its role in osmotic and salt stress responses in birch. Notably, under osmotic and salt stress, Bp-miR408a contributed to osmotic and salt tolerance sensitivity by mediating various physiological changes, such as increases in reactive oxygen species accumulation, osmoregulatory substance contents and Na+ accumulation. Additionally, molecular analysis provided evidence of the in vivo targeting of BpBCP1 (blue copper protein) transcripts by Bp-miR408a. The overexpression of BpBCP1 in birch enhanced osmotic and salt tolerance by increasing the antioxidant enzyme activity, maintaining cellular ion homeostasis and decreasing lipid peroxidation and cell death. Thus, we reveal a Bp-miR408a-BpBCP1 regulatory module that mediates osmotic and salt stress responses in birch.

Rapid Detection of Microparticles Using a Microfluidic Resistive Pulse Sensor Based on Bipolar Pulse-Width Multiplexing.

Rapid and accurate analysis of micro/nano bio-objects (e.g., cells, biomolecules) is crucial in clinical diagnostics and drug discovery. While a traditional resistive pulse sensor can provide multiple kinds of information (size, count, surface charge, etc.) about analytes, it has low throughput. We present a unique bipolar pulse-width, multiplexing-based resistive pulse sensor for high-throughput analysis of microparticles. Signal multiplexing is enabled by exposing the central electrode at different locations inside the parallel sensing channels. Together with two common electrodes, the central electrode encodes the electrical signal from each sensing channel, generating specific bipolar template waveforms with different pulse widths. Only one DC source is needed as input, and only one combined electrical output is collected. The combined signal can be demodulated using correlation analysis and a unique iterative cancellation scheme. The accuracy of particle counting and sizing was validated using mixtures of various sized microparticles. Results showed errors of 2.6% and 6.1% in sizing and counting, respectively. We further demonstrated its accuracy for cell analysis using HeLa cells.

The Complete Chloroplast Genome Sequence of Staphylea holocarpa (Staphyleaceae).

Staphylea holocarpa (Hemsley 1895) is an ornamental deciduous shrub or tree in the family Staphyleaceae. As the shortage of the wild resources, S. holocarpa is also a rare plant. The revelation of the species origin and evolution progress and the relation. Therefore, the S. holocarpa complete chloroplast genome sequence was completed and characterized by de novo assembly. The cp genome length of S. holocarpa was 160,461 bp and it has a typical quadripartite structure, consisted of an 89,760 bp large single-copy region and a 18,639 bp small single-copy region, which were divided by two inverted repeat regions of 26,031 bp. After genome annotation, it comes to 130 genes that were predicted, which includes 85, 8, and 37 encoded proteins, rRNA, and tRNA, respectively. A phylogenetic analysis has shown that the S. holocarpa cp genome is related to the Staphylea trifolia. This work will be useful for further population genomic and phylogenetic studies of S. holocarpa.

Recent Advances in Biomolecular Detection Based on Aptamers and Nanoparticles.

The fast, accurate detection of biomolecules, ranging from nucleic acids and small molecules to proteins and cellular secretions, plays an essential role in various biomedical applications. These include disease diagnostics and prognostics, environmental monitoring, public health, and food safety. Aptamer recognition (DNA or RNA) has gained extensive attention for biomolecular detection due to its high selectivity, affinity, reproducibility, and robustness. Concurrently, biosensing with nanoparticles has been widely used for its high carrier capacity, stability and feasibility of incorporating optical and catalytic activity, and enhanced diffusivity. Biosensors based on aptamers and nanoparticles utilize the combination of their advantages and have become a promising technology for detecting of a wide variety of biomolecules with high sensitivity, reliability, specificity, and detection speed. Via various sensing mechanisms, target biomolecules have been quantified in terms of optical (e.g., colorimetric and fluorometric), magnetic, and electrical signals. In this review, we summarize the recent advances in and compare different aptamer-nanoparticle-based biosensors by nanoparticle types and detection mechanisms. We also share our views on the highlights and challenges of the different nanoparticle-aptamer-based biosensors.

Genome-wide identification and expression profiling of the bZIP gene family in Betula platyphylla and the functional characterization of BpChr04G00610 under low-temperature stress.

The basic leucine zipper (bZIP) gene, which plays a significant role in the regulation of tolerance to biotic/abiotic stresses, has been characterized in many plant species. Betula platyphylla is a significant afforestation species. To elucidate the stress resistance mechanism of birch, previous studies identified some stress resistance genes. However, the genome-wide identification and characterization of bZIP gene family in the birch have not been reported. Here, the 56 BpbZIP genes were identified and classified into 13 groups in birch. Cis-element analysis showed that the promoters of 56 family genes contained 108 elements, of which 16 were shared by 13 groups. There were 8 pairs of fragment repeats and 1 pair of tandem repeats, indicating that duplication may be the major reason for the amplification of the BpbZIP gene family. Tissue-specific of BpbZIP genes showed 18 genes with the highest expression in roots, 15 in flowers, 11 in xylem and 9 in leaves. In addition, five differentially expressed bZIP genes were identified from the RNA-seq data of birch under low-temperature stress, and the co-expressed differentially expressed genes were further screened. The analysis of gene ontology (GO) enrichment of each co-expression regulatory network showed that they were related to membrane lipids and cell walls. Furthermore, the transient overexpression of BpChr04G00610 decreased the ROS scavenging ability of birch under low-temperature stress, suggesting that it may be more sensitive to low-temperature. In conclusion, this study provides a basis for the study of the function of BpbZIP genes.

Cell Surface Charge Mapping Using a Microelectrode Array on ITO Substrate.

Many cellular functions are regulated by cell surface charges, such as intercellular signaling and metabolism. Noninvasive measurement of surface charge distribution of a single cell plays a vital role in understanding cellular functions via cell membranes. We report a method for cell surface charge mapping via photoelectric interactions. A cell is placed on an array of microelectrodes fabricated on a transparent ITO (indium tin oxide) surface. An incident light irradiates the ITO surface from the backside. Because of the influence of the cell surface charge (or zeta potential), the photocurrent and the absorption of the incident light are changed, inducing a magnitude change of the reflected light. Hence, the cell surface charge distribution can be quantified by analyzing the reflected light intensity. This method does not need physical or chemical modification of the cell surface. We validated this method using charged microparticles (MPs) and two types of cells, i.e., human dermal fibroblast cells (HDFs) and human mesenchymal stem cells (hMSC). The measured average zeta potentials were in good agreement with the standard electrophoresis light scattering method.

Two-Dimensional Nitrogen-Doped Carbon Nanosheets Derived from g-C3N4 /ZIF-8 for Solid-Phase Microextraction in Exhalation of Esophageal Cancer Patients.

Here, two-dimensional (2D) nitrogen-doped carbon nanosheets (CNSs) were prepared through carbonizing MOFs (ZIF-8) in-situ grown using graphitic carbon nitride (g-C3N4) as a template. The developed ZIF-8 CNS was then used as solid-phase microextraction (SPME) fiber coating for beneficiation of five biomarkers in exhalation of patients with esophageal cancer and in gas chromatography-mass spectrometry (GC-MS) for determination. The ZIF-8 CNS fiber exhibits satisfactory enrichment factors (3490-5631), wide linearity (5-1000 µg L-1), and low detection limits (0.26-0.96 µg L-1). The relative standard deviations (RSDs) for six replicate extractions of the same ZIF-8 CNS fiber were between 2.0-3.9% (intra-day) and 2.8-5.2% (inter-day). The reproducibility of three fibers prepared by the same approach was in the range 6.8-12.3% (RSD). The developed ZIF-8 CNS fiber can persist in 120 SPME cycles with no prominent loss of extraction efficiency and precision. The high enrichment factors of the 2D ZIF-8 CNS coatings are attributed to the high specific surface area, ultrathin thickness, and nano-pore or interlayer channels; moreover, nitrogen doping also endows the π system with a strong electron absorption ability, which will enhance the π-π interaction between the ZIF-8 CNS and the aromatic ring. Ultimately, the self-made ZIF-8 CNS-coated SPME fiber was applied to the analysis of exhaled breath samples. The recoveries of spiked analytes are between 84 and 105%.

[Analysis of pathogenic gene variant in two children with Treacher-Collins syndrome].

OBJECTIVE: To explore the clinical and genetic characteristics of two children with a clinical diagnosis of Treacher Collins syndrome (TCS). METHODS: Whole-exome sequencing was used to screen potential variants in the two children. Confirmation of suspected variants was performed through Sanger sequencing, multiplex ligation dependent probe amplification and real-time PCR in probands and their parents. RESULTS: A heterozygous deletion variant, c.4357_4360delGAAA, was detected in case one, while was de novo and verified by Sanger sequencing. The variant was classified as pathogenic (PVS1 +PM2+PM6) according to ACMG guideline. The heterozygous deletion of exon 1-7 was seen in the same gene in case 2, which MLPA verified as heterozygous deletion of exon 1-6. This deletion was inherited from the father with a normal phenotype, and the father's TCOF1 gene was suspected to be chimeric heterozygous deletion of exon 1-6 verified by MLPA. CONCLUSION: The identified variants in the TCOF1 gene probably underlie the two cases of TCS. There was no apparent correlation between genotype and phenotype. In addition, it shows a high interfamilial variability ranging from normal to full presentation of TCS. Genetic detection provided clinical diagnosis and genetic counselling for TCS patients.

Nutritional Value, Fermentation Characteristics and In Vitro Degradability of Whole Wheat Hay Harvested at Three Stages of Maturity.

The nutritional value of whole crop wheat hay (WCWH) harvested at different maturation stages are different, and its feeding effects on dairy cows have not been thoroughly evaluated. In this study, the in vitro digestibility of whole wheat (Nongda 22) hay harvested during the flowering, late milk and dough stages were evaluated using batch culture technique. The neutral detergent fiber (NDF) and acid detergent fiber (ADF) contents of whole wheat hay decreased by 35.5% and 40.4%, respectively, whereas the non-fibrous carbohydrates (NFC) content increased by 50.3% in WCWH harvested during the dough stage as compared to the flowering stage (p < 0.01). The pH of the fermentation liquid and acetate to propionate ratio was greatest in the wheat harvested during the flowering stage and lowest during the dough stage (p = 0.03), whereas the volatile fatty acid (VFA) concentration was greatest during the dough stage and lowest during the flowering stage (p < 0.01). The dry matter loss (DML) was 9.6% and 6.2% greater (p < 0.01) during the late milk stage than in the flowering or dough stages, and the NDF loss (NDFL; p = 0.01) and ADF loss (ADFL; p < 0.01) was greater in both the flowering and late milk stages. In conclusion, though the content of NDF was lower in the dough stage, and the starch to NFC ratio was greater, we determined that the optimal harvest stage should be the late milk stage due to the greater dry matter digestibility, the relatively greater NFC content and the shorter planting days.

Neuropathologic damage induced by radiofrequency ablation at different temperatures.

OBJECTIVE: To explore the molecular mechanism of neuropathologic damage induced by radiofrequency ablation at different temperatures. METHODS: This is basic research, and 36 SD rats were used to construct the neuropathological injury model. The rats were subjected to radiofrequency stimulation at different temperatures and were divided into 6 groups according to the temperature injury: 42°, 47°, 52°, 57°, 62°, and 67°C groups. Conduction time, conduction distance, and nerve conduction velocity were recorded after temperature injury. HE-staining was used to observe the histopathological morphology of the sciatic nerve. The expression of SCN9A, SCN3B, and NFASC protein in sciatic nerve tissue were detected by western blot. RESULTS: With the increase in temperature, nerve conduction velocity gradually decreased, and neurons were damaged when the temperature was 67°C. HE-staining showed that the degrees of degeneration of neurons in rats at 47°, 52°, 57°, 62°, and 67°C were gradually increased. The expression of SCN9A, SCN3B protein in 57°, 62°, 67°C groups were much higher than that of NC, 42°, 47°, 52°C groups. However, the expression of NFASC protein in 57°, 62°, 67°C groups was much lower than that of the NC, 42°, 47°, 52°C groups. CONCLUSION: There was a positive correlation between temperature caused by the radiofrequency stimulation to neuropathological damage. The mechanism is closely related to the expression of SCN9A, SCN3B, and NFASC protein in nerve tissue caused by heat transfer injury.

Ultrasensitive detection of small biomolecules using aptamer-based molecular recognition and nanoparticle counting.

Detection of small biomolecules is critical for understanding molecular mechanisms in biological systems and performing in vitro diagnosis in clinics. Current antibody based detection methods face large challenges in detecting small biomolecules at low concentrations. We report a new method for detecting small biomolecules based on molecular recognition and nanoparticle (NP) counting. Aptamer-functionalized NPs are attached to complementary sequence (CS)-conjugated microparticle (MP) carriers. In the presence of target small biomolecules at ultra low concentrations, NPs would be released from the MP carriers. Coupled with a resistive pulse sensor (RPS) using a micropore that counts the released NPs, this method can measure the concentrations of target biomolecules at low concentrations with high sensitivity and high throughput. Adenosine was used as a model to demonstrate the feasibility of this method. It is demonstrated that this method can detect a wide range of adenosine concentrations with a low detection limit of 0.168 nM, which is 10 times lower than that of the ELISA kit. With its simple structure, high sensitivity, and high reproducibility, this detection method holds great potential for the ultrasensitive detection of low abundance small biomolecules.

Neuropathologic damage induced by radiofrequency ablation at different temperatures

Abstract Objective To explore the molecular mechanism of neuropathologic damage induced by radiofrequency ablation at different temperatures. Methods This is basic research, and 36 SD rats were used to construct the neuropathological injury model. The rats were subjected to radiofrequency stimulation at different temperatures and were divided into 6 groups according to the temperature injury: 42°, 47°, 52°, 57°, 62°, and 67°C groups. Conduction time, conduction distance, and nerve conduction velocity were recorded after temperature injury. HE-staining was used to observe the histopathological morphology of the sciatic nerve. The expression of SCN9A, SCN3B, and NFASC protein in sciatic nerve tissue were detected by western blot. Results With the increase in temperature, nerve conduction velocity gradually decreased, and neurons were damaged when the temperature was 67°C. HE-staining showed that the degrees of degeneration of neurons in rats at 47°, 52°, 57°, 62°, and 67°C were gradually increased. The expression of SCN9A, SCN3B protein in 57°, 62°, 67°C groups were much higher than that of NC, 42°, 47°, 52°C groups. However, the expression of NFASC protein in 57°, 62°, 67°C groups was much lower than that of the NC, 42°, 47°, 52°C groups. Conclusion There was a positive correlation between temperature caused by the radiofrequency stimulation to neuropathological damage. The mechanism is closely related to the expression of SCN9A, SCN3B, and NFASC protein in nerve tissue caused by heat transfer injury.

CircularLRRC7 is a Potential Tumor Suppressor Associated With miR-1281 and PDXP Expression in Glioblastoma.

Circular RNAs (circRNAs) are usually enriched in neural tissues, yet about 80% circRNAs have lower expression in gliomas relative to normal brains, highlighting the importance of circRNAs as tumor suppressors. However, the clinical impact as well as the pathways regulated by the tumor-suppressive circRNAs remain largely unknown in glioblastoma (GBM). Through bioinformatic analysis followed by experimental validation, we found that hsa_circ_0114014 (circLRRC7) was dramatically down-regulated in GBM when compared with normal brain tissues (p < 0.0001). GBM patients with a lower circLRRC7 expression had poorer progression-free survival (PFS, p < 0.05) and overall survival (OS, p < 0.05). Analyses of the predicted target miRNAs of circLRRC7 in CSCD and CRI databases, in combination with the miRNA expression data in GBMs and normal brains from GSE database, revealed miR-1281 as a potential downstream target of circLRRC7. Subsequently, the target genes of hsa-mir-1281 were predicted by TargetScan, miRDB and miRNATAR databases. Intersection analysis and correlation test indicated that PDXP was a potential target of miR-1281. In summary, circLRRC7 may be a tumor suppressor that associated with miR-1281 and PDXP expression in GBM, which may provide novel therapeutic targets for GBM treatment.

Mapping Surface Charge Distribution of Single-Cell via Charged Nanoparticle.

Many bio-functions of cells can be regulated by their surface charge characteristics. Mapping surface charge density in a single cell's surface is vital to advance the understanding of cell behaviors. This article demonstrates a method of cell surface charge mapping via electrostatic cell-nanoparticle (NP) interactions. Fluorescent nanoparticles (NPs) were used as the marker to investigate single cells' surface charge distribution. The nanoparticles with opposite charges were electrostatically bonded to the cell surface; a stack of fluorescence distribution on a cell's surface at a series of vertical distances was imaged and analyzed. By establishing a relationship between fluorescent light intensity and number of nanoparticles, cells' surface charge distribution was quantified from the fluorescence distribution. Two types of cells, human umbilical vein endothelial cells (HUVECs) and HeLa cells, were tested. From the measured surface charge density of a group of single cells, the average zeta potentials of the two types of cells were obtained, which are in good agreement with the standard electrophoretic light scattering measurement. This method can be used for rapid surface charge mapping of single particles or cells, and can advance cell-surface-charge characterization applications in many biomedical fields.

The Antidepressant Effects of Resveratrol are Accompanied by the Attenuation of Dendrite/Dendritic Spine Loss and the Upregulation of BDNF/p-cofilin1 Levels in Chronic Restraint Mice.

Depression afflicts more than 300 million people worldwide, but there is currently no universally effective drug in clinical practice. In this study, chronic restraint stress (CRS)-induced mice depression model was used to study the antidepressant effects of resveratrol and its mechanism. Our results showed that resveratrol significantly attenuated depression-like behavior in mice. Consistent with behavioral changes, resveratrol significantly attenuated CRS-induced reduction in the density of dendrites and dendritic spines in both hippocampus and medial prefrontal cortex (mPFC). Meanwhile, in hippocampus and mPFC, resveratrol consistently alleviated CRS-induced cofilin1 activation by increasing its ser3 phosphorylation. In addition, cofilin1 immunofluorescence distribution in neuronal inner peri-membrane in controls, and cofilin1 diffusely distribution in the cytoplasm in CRS group were common in hippocampus. However, the distribution of cofilin1 in mPFC was reversed. Pearson's correlation analysis revealed that there was a significant positive correlation found between the sucrose consumption in sucrose preference test and the dendrite density in multiple sub-regions of hippocampus and mPFC, and a significant negative correlation between the immobility time in tail suspension test and the dendrite/dendritic spine density in several different areas of hippocampus and mPFC. P-cofilin1 was significantly positively correlated with the overall dendritic spine density in mPFC as well as with the overall dendrite density or BDNF in the hippocampus. Our results suggest that the BDNF/cofilin1 pathway, in which cofilin1 may be activated in a brain-specific manner, was involved in resveratrol's attenuating the dendrite and dendritic spine loss and behavioral abnormality.

A Microfluidic Sensor for Continuous, in Situ Surface Charge Measurement of Single Cells.

Cell surface charge has been recognized as an important cellular property. We developed a microfluidic sensor based on resistive pulse sensing to assess surface charge and sizes of single cells suspended in a continuous flow. The device consists of two consecutive resistive pulse sensors (RPSs) with identical dimensions. Opposite electric fields were applied on the two RPSs. A charged cell in the RPSs was accelerated or decelerated by the electric fields and thus exhibited different transit times passing through the two RPSs. The cell surface charge is measured with zeta potential that can be quantified with the transit time difference. The transit time of each cell can be accurately detected with the width of pulses generated by the RPS, while the cell size can be calculated with the pulse magnitude at the same time. This device has the ability to detect surface charges and sizes of individual cells with high tolerance in cell types and testing solutions compared with traditional electrophoretic light scattering methods. Three different types of cells including HeLa cancer cells, human dermal fibroblast cells, and human umbilical vein endothelial cells (HUVECs) were tested with the sensor. Results showed a significant difference of zeta potentials between HeLa cells and fibroblasts or HUVECs. In addition, when HeLa cells were treated with various concentrations of glutamine, the effects on cancer cell surface charge were detected. Our results demonstrated the great potential of using our sensor for cell type sorting, cancer cell detection, and cell status analysis.