Induction of pancreatic duct cells of neonatal rats into insulin-producing cells with fetal bovine serum: A natural protocol and its use for patch clamp experiments

AIM： To induce the pancreatic duct cells into endocrine cells with a new natural protocol for electrophysiological study. METHODS： The pancreatic duct cells of neonatal rats were isolated, cultured and induced into endocrine ceils with 15% fetal bovine serum for a period of 20 d. During this period, insulin secretion, MTT value, and morphological change of neonatal and adult pancreatic islet cells were comparatively investigated. Pancreatic β-cells were identified by morphological and electrophysiological characteristics, while ATP sensitive potassium channels （KATP）, voltage-dependent potassium channels （Kv）, and voltage-dependent calcium channels （KcA） in β-cells were identified by patch clamp technique. RESULTS： After incubation with fetal bovine serum, the neonatal duct cells budded out, changed from duct-like cells into islet clusters. In the first 4 d, MTT value and insulin secretion increased slowly （MTT value from 0.024 ±0.003 to 3.028±0.003, insulin secretion from 2.6±0.6 to 3.1±0.8 mIU/L）. Then MTT value and insulin secretion increased quickly from d 5 to d 10 （MTT value from 0.028 ±0.003 to 0.052±0.008, insulin secretion from 3.1±0.8 to 18.3±2.6 mIU/L）, then reached high plateau （MTT value 〉0.052±0.008, insulin secretion 〉18.3±2.6 mIU/L）. In contrast, for the isolated adult pancreatic islet cells, both insulin release and MTT value were stable in the first 4 d （MTT value from 0.029±0.01 to 0.031±0.011, insulin secretion from 13.9±3.1 to 14.3±3.3 mIU/L）, but afterwards they reduced gradually （MTT value 〈0.031 ±0.011, insulin secretion 〈8.2±1.5 mIU/L）, and the pancreatic islet cells became dispersed, broken or atrophied correspondingly. The differentiated neonatal cells were identified as pancreatic islet cells by dithizone staining method, and pancreatic β-cells were further identified by both morphological features and electrophysiological characteristics, i.e. the existence of recording currents from KATP, Kv, and KCA. CONCLUSION： Islet cells differentiated from neonatal pancreatic duct cells with the new natural protocol are more advantageous in performing patch clamp study over the isolated adult pancreatic islet cells.

A patch-clamp study on human sperm Cl^- channel reassembled into giant liposome

Aim: To record the single-channel currents and characterize the electrophysiological properties of the Cl^- channels inhuman sperm membrane. Methods: The membrane proteins extracted from the human sperm were reassembled intoliposome bilayer, and the liposomes were fused into giant liposomes with a diameter more than 10μm by dehydration-rehydration procedure. The giant liposomes were used to study the Cl^- channel activities by patch-clamp technique.Results: By patch clamping the giant liposome in an asymmetric NMDG (N-methyl-D-glucamine)-Cl (bath 100//pipette 200 mmol/L) solution system, three kinds of single-channel events with unit conductances of (74.1 ± 8.3) pS,(117.0±5.7) pS and (144.7±4.5) pS, respectively, were detected. Their activities were voltage-dependent and allwere blocked by SITS (4-acetamido-4'-isothiocyanato-stilbene-2', 2'-disulfonic acid) in a concentration-dependentmanner. By constructing the open and close dwell time distribution histograms and then fitting them with exponentialfunction, two time constants were obtained in both the open and the close states. The burst activity and conductancesubstate of the channels were observed. Conclusion; There exist three kinds of Cl^- channels with different conduc-tance in human sperm membrane at least. (Asian J Androl 2001 Sep; 3: 185 - 191)

In vivo patch clamp recording technique in the study of neurophysiology

The patch clamp recording technique in vivois a blind patch clamp recording methods to record the current of the spinal or cereral neurons of anaes：hesia （ or awake） animals. This technique can be used to study the synaptic function and plasticity in central nervous system in vivoin order to understand the physiological properties of the ion channels from an integrated point of view. The advantage of this technique have already presented itself in the study of the synaptic transmission and nervous network. Nowadays, in vivo patch whole-cell recording technique in combination with other techniques is becoming a common method in the research fields.

Action Mechanism Research of Lanthanons to Slow Vacuolar Ion Channels in Raphanus Satirus L.(Xinlimei) Radish by Patch-Clamp

We used whole-vacuolar patch-clamp recording mode to study the action mechanism of La3+ to Slow Vacuolar (SV) channels for the first time. We recorded SV channel currents of Xinlimei (Raphanus satirus L.) vacuolars. The minimum activation potentials of voltage-dependent SV channels tied in 25+/-5 mV. The increase in cytoplasmic Ca2+ led to enhancement of SV-type currents. It was found that the threshold potential of activation shifted towards more depolarized values whenever cytoplasmic Ca2+ was increased. When 10(-10) mol/L free La3+ was added to the bath, SV-type current was suppressed by 60 similar to 75%. These data showed La3+ reduced ion permeabilities of Xinlimei root vacuolar membrane.

Study of Oligonucleotide Fixation on Bilayer Lipid Membranes by Patch-clamp Pipette Support

One kind of novel BLMs was fabricated by patch-clamp pipette technology characterized in considerably sensitive to changes of electrochemical parameters.Detectiye currents and voltage presented linear relationship when BLMs was formed and it could be confirmed by Gramicidin method.Ion current was increased by dihexyl (C_ (12)) modified ssDNA fixed on the BLMs and also indicated linear relationship to ssDNA's concentration due to the interaction of (C_ 12)-ssDNA and BLMs.Further more,the regression equations were different from BLMs fixed with ssDNA probe and a blank control BLM in the same experimental conditions.The ssDNA probe was successfully fixed on patch-clamp pipette supported-BLMs.Based on our studies,a biosensor with reactive element of patch-clamp pipette-supported BLMs has been established.

The Reconstructing of Low Signal-noise Ratio Single Ion Channel Signal from Patch-clamp Recordings Sampled in the Colored Background Noise

The single ion channel signal is an ionic current that can be recorded by the patch clamp technique. Hidden Markov model (HMM) algorithm has been used to convert the low signal noise ratio (SNR) noisy recording into an idealized quantal one in the case of white background noise. The traditional HMM algorithm is extended and adapted to the colored background noise. A new algorithm called EHMM (Extended HMM) algorithm is proposed, and mainly validated by simulation. Results show that it’s effective.

Feasibility and limitation of patch-clamp recordings on neonatal rat cardiac ventricular slices

To attempt to the feasibility of whole-cell patch-clamp recordings on the cardiac ventricular slices of newborn (P3-P7) Sprague-Dawley (S-D) rats and find a good substitute of the single cardiac myocytes with enzymatic treatment. High resistance seals (】1 GΩ) could be obtained in the cardiac ventricle tissue on this preparation without enzymatic treatment. Then, the cell-attached and whole-cell patch-clamp techniques can be achieved in the thin (200μm) cardiac slices. Averaged sodium current (n=ll cells) was recorded in cell-attached mode, displayed similar features to that previously reported from isolated rat ventricular myocytes. The outward potassium current, Hyperpolarization-activated cation channel or If channel (HCN channel) and action potential (AP) were recorded in whole-cell configuration (n=2 cells) and the similar properties can be seen from the cardiac slices. Cell-attached mode is a stable and reliable mode to record the ion treatment. Resting potential for cardiac slice, measured from Current-clamp recording in whole-cell mode, is about -50 - - 70 mV. The resting potential value from the cardiac slice has similar property except that it is positive to the isolated cardiomyocytes by enzymes. Application of patch-clamp techniques to cardiac slices allows single channel recordings without complicated procedures of cell isolation. Moreover, possible alteration of channel properties caused by proteolytic enzymes can be avoided. In this paper, whole-cell patch-clamp recordings could be achieved on the cardiac slice and we affirmed the feasibility and values of the both recording modes on it. At the same time, there is difficulty and limitation of the application of whole-cell patch-clamp on the cardiac slice for the existence of large mount of connective tissue even in the newborn rats.

基于PatchTracker的对抗补丁防御算法

基于深度神经网络的目标检测技术已经广泛应用于各领域,然而,通过对抗补丁攻击在图像中添加局部扰动,以此来误导深度神经网络,对基于目标检测技术的视觉系统构成了严重威胁。针对这一问题,利用对抗补丁和图像背景的语义差异性,提出了一种基于PatchTracker的对抗补丁防御算法,该算法由上游补丁检测器与下游数据增强模块组成。上游补丁检测器使用带有注意力机制的YOLOV5(you only look once-v5)确定对抗补丁所在位置,有助于提高对小尺度对抗补丁的检测精度;将检测区域用合适的像素值覆盖以抹除对抗补丁,上游补丁检测器不仅能够有效降低对抗样本的攻击性,而且不依赖大规模的训练数据;下游数据增强模块通过改进模型训练范式,提高下游目标检测器的鲁棒性;将抹除补丁后的图像输入经过数据增强的下游YOLOV5目标检测模型。在公开的TT100K交通标志数据集上进行了交叉验证,实验表明,与未采取防御措施相比,所提算法能够有效防御多种类型的通用对抗补丁攻击,在检测对抗补丁图像时的mAP(mean average precision)提高65%左右,有效地改善了小尺度对抗补丁的漏检情况。与现有算法比较,所提算法有效提高了神经网络在检测对抗样本时的准确率。此外,所提算法不涉及下游模型结构的修改,具有良好的兼容性。

Scheme Based on Multi-Level Patch Attention and Lesion Localization for Diabetic Retinopathy Grading

Early screening of diabetes retinopathy(DR)plays an important role in preventing irreversible blindness.Existing research has failed to fully explore effective DR lesion information in fundus maps.Besides,traditional attention schemes have not considered the impact of lesion type differences on grading,resulting in unreasonable extraction of important lesion features.Therefore,this paper proposes a DR diagnosis scheme that integrates a multi-level patch attention generator(MPAG)and a lesion localization module(LLM).Firstly,MPAGis used to predict patches of different sizes and generate a weighted attention map based on the prediction score and the types of lesions contained in the patches,fully considering the impact of lesion type differences on grading,solving the problem that the attention maps of lesions cannot be further refined and then adapted to the final DR diagnosis task.Secondly,the LLM generates a global attention map based on localization.Finally,the weighted attention map and global attention map are weighted with the fundus map to fully explore effective DR lesion information and increase the attention of the classification network to lesion details.This paper demonstrates the effectiveness of the proposed method through extensive experiments on the public DDR dataset,obtaining an accuracy of 0.8064.

Patch-Locator:一种基于排序学习的开源软件漏洞补丁定位方法

日益增多的开源软件漏洞对软件安全带来了巨大的风险,补丁在应对这一风险的过程中扮演了非常重要的角色.不幸的是,尽管大部分漏洞的补丁在被披露前就已经开发完毕,但仅有部分补丁会随漏洞同步公开.现有的研究发现了漏洞与其补丁之间存在一定的相关性,并基于这些相关性特征对提交进行了排序,以定位漏洞的补丁,但仍旧存在漏洞数据部分缺失、定位准确率不佳等问题.本文提出了Patch-Locator,一种新的基于排序学习的补丁定位方法,通过扩展漏洞数据源对漏洞数据进行补充,并根据漏洞与补丁文本的相似性、漏洞产生的原因和导致的结果等更能反映漏洞与补丁间关联的因素提取了更具有针对性的相关性特征,并使用LambdaMart排序学习模型对提交基于其具有的相关性特征进行排序以定位安全补丁.本文用来自10个开源软件项目的1669个漏洞来评估Patch-Locator.实验结果表明,Patch-Locator的Recall@1指标为92.22%,Recall@5指标为95.51%,Manual Effort@5指标为1.2455,均优于现有方法.

Comparison of nonlinear modeling methods for the composite rubber clamp

The cubic stiffness force model(CSFM)and Bouc-Wen model(BWM)are introduced and compared innovatively.The unknown coefficients of the nonlinear models are identified by the genetic algorithm combined with experiments.By fitting the identified nonlinear coefficients under different excitation amplitudes,the nonlinear vibration responses of the system are predicted.The results show that the accuracy of the BWM is higher than that of the CSFM,especially in the non-resonant region.However,the optimization time of the BWM is longer than that of the CSFM.

Mechanobiomaterials:Mechanics-Guided Design of Epicardial Patch for Treating

In recent years,the field of mechanomaterials has emerged at the interface of mechanics,materials science,biology,medicine and data science,where materials are proactively designed or programmed to achieve targeted functionalities by leveraging the fundamental mechanics principles and force-geometry-property relationships.In the biological context,one may likewise introduce mechanobiomaterials as a field with the following goals:(1)proactive design or programming of materials for precisely mediating biomechanical environment of living systems for tissue repair/restoration;(2)proactive control/programming of living systems themselves by an external field via force-structure-function relationships.Here,we will discuss an example of research in mechanobiomaterials on using mechanics to guide the design of acellular epicardial patches for the treatment of myocardial infarction.This technology aims to employ a biocompatible material patch to help reverse left ventricular remodeling and restore heart function after myocardial infarction by increasing the mechanical integrity of damaged heart tissues.However,its application is currently limited by widely scattered therapeutic efficacy.Here,we develop a biomechanics-based simulation platform that allows us to test,design and optimize the performance of an epicardial patch.We show that the widely scattered therapeutic efficacy of this technology can be attributed to a“pre-strain sensitivity”caused by attaching an elastic patch to a dynamically beating heart.To mitigate this challenge,we introduce a viscoelastic epicardial patch,designed at the so-called‘gel point’of the material,that effectively accommodates the cyclic deformation of the myocardium.This then leads to the fabrication and experimentally validated epicardial patch that outperforms all existing ones in restoring heart function after both acute and subacute myocardial infarction in rats.Our study also demonstrates the potential of employing viscoelastic interfaces for better integration of synthetic materials with biological tissues.

Effect of an Airbag-selective Portal Vein Blood Arrester on the Liver after Hepatectomy:A New Technique for Selective Clamping of the Portal Vein

Objective:A novel technique was explored using an airbag-selective portal vein blood arrester that circumvents the need for an intraoperative assessment of anatomical variations in patients with complex intrahepatic space-occupying lesions.Methods:Rabbits undergoing hepatectomy were randomly assigned to 4 groups:intermittent portal triad clamping(PTC),intermittent portal vein clamping(PVC),intermittent portal vein blocker with an airbag-selective portal vein blood arrester(APC),and without portal blood occlusion(control).Hepatic ischemia and reperfusion injury were assessed by measuring the 7-day survival rate,blood loss,liver function,hepatic pathology,hepatic inflammatory cytokine infiltration,hepatic malondialdehyde levels,and proliferating cell nuclear antigen levels.Results:Liver damage was substantially reduced in the APC and PVC groups.The APC animals exhibited transaminase levels similar to or less oxidative stress damage and inflammatory hepatocellular injury compared to those exhibited by the PVC animals.Bleeding was significantly higher in the control group than in the other groups.The APC group had less bleeding than the PVC group because of the avoidance of portal vein skeletonization during hepatectomy.Thus,more operative time was saved in the APC group than in the PVC group.Moreover,the total 7-day survival rate in the APC group was higher than that in the PTC group.Conclusion:Airbag-selective portal vein blood arresters may help protect against hepatic ischemia and reperfusion injury in rabbits undergoing partial hepatectomy.This technique may also help prevent liver damage in patients requiring hepatectomy.

Formation and ecological response of sand patches in the protection system of Shapotou section of the Baotou-Lanzhou railway,China

The development of bare patches typically signifies a process of ecosystem degradation.Within the protection system of Shapotou section of the Baotou-Lanzhou railway,the extensive emergence of bare sand patches poses a threat to both stability and sustainability.However,there is limited knowledge regarding the morphology,dynamic changes,and ecological responses associated with these sand patches.Therefore,we analyzed the formation and development process of sand patches within the protection system and its effects on herbaceous vegetation growth and soil nutrients through field observation,survey,and indoor analysis methods.The results showed that sand patch development can be divided into three stages,i.e.,formation,expansion,and stabilization,which correspond to the initial,actively developing,and semi-fixed sand patches,respectively.The average dimensions of all sand patch erosional areas were found to be 7.72 m in length,3.91 m in width,and 0.32 m in depth.The actively developing sand patches were the largest,and the initial sand patches were the smallest.Throughout the stage of formation and expansion,the herbaceous community composition changed,and the plant density decreased by more than 50.95%.Moreover,the coverage and height of herbaceous plants decreased in the erosional area and slightly increased in the depositional lobe;and the fine particles and nutrients of soils in the erosional area and depositional lobe showed a decreasing trend.In the stabilization phases of sand patches,the area from the inlet to the bottom of sand patches becomes initially covered with crusts.Vegetation and 0-2 cm surface soil condition improved in the erosional area,but this improvement was not yet evident in the depositional lobe.Factors such as disturbance,climate change,and surface resistance to erosion exert notable influences on the formation and dynamics of sand patches.The results can provide evidence for the future treatment of sand patches and the management of the protection system of Shapotou section of the Baotou-Lanzhou railway.

Do brood parasitic Common Cuckoos develop brood patches during the breeding season?

Many birds develop brood patches on their ventral apterium under hormonal regulation to effectively transmit body heat to eggs during incubation.The developed patch has several characteristics,including de-feathering,vascularization,and edema.However,little is known about whether avian brood parasites that do not incubate their eggs exhibit brood patch development during the breeding season.In this study,we measured the size of the ventral apterium in 114 Common Cuckoos(Cuculus canorus)captured in the field throughout the breeding season and examined the appearance of the ventral apterium to confirm the development of brood patches.We also examined whether morphological traits and various factors correlated with the size of the ventral apterium(sternal apterium and abdominal apterium)and how it changed during the breeding season.We found no clear signs of brood patch development in Common Cuckoos captured throughout the breeding season,indicating that they likely did not develop brood patches on the ventral apterium during this period.We also found that ventral apterium size was positively correlated with wing length and body weight.In addition,Common Cuckoos with newly growing feathers on the boundary of the ventral apterium with pteryla were frequently observed as seasons progressed to the end.In conclusion,Common Cuckoos exhibit neither brood patch development nor the vestigial characteristics of ancestral brood patches,except for the growth of feathers on boundary the ventral apterium.Further studies examining brood patches across different groups of avian brood parasites are valuable for enhancing our understanding of the developmental and physiological adaptations of avian brood parasites.

Treatment of 198 Cases of Thyroid Nodules with Sanying Capsule Combined with Xiaoying Patch

[Objectives]To observe the clinical efficacy of Sanying capsule combined with Xiaoying Patch in treating thyroid nodule(TN).[Methods]Two groups were treated similarly,with 200 cases in the control group undergoing basic treatment for 12 weeks and 198 cases in the observation group receiving Sanying capsule combined with Xiaoying patches for the same duration.The clinical symptoms,number of nodules,diameter of the largest nodule,and maximum reduction of nodules were observed before and after treatment.A control analysis was performed,and the underlying mechanisms were explored.[Results]The primary symptoms of the observation group exhibited a more favorable improvement than those of the control group.Additionally,the number of nodules decreased,the diameter of the largest nodule decreased,and the maximum reduction of nodules decreased in both groups following treatment.However,the observation group demonstrated a more pronounced improvement than the control group(P<0.05).[Conclusions]The combination of Sanying capsule and Xiaoying patch has been demonstrated to be an effective treatment for TN,with a high degree of reliability in terms of safety.

Ginger oil-loaded transdermal adhesive patch treats post-traumatic stress disorder

Objective:To find a viable alternative to reduce the number of doses required for the patients with post-traumatic stress disorder(PTSD),and to improve efficacy and patient compliance.Methods: In this study,we used ginger oil,a phytochemical with potential therapeutic properties,to prepare ginger oil patches.High-performance liquid chromatography(HPLC)was used to quantify the main active component of ginger oil,6-gingerol.Transdermal absorption experiments were conducted to optimize the various pressure-sensitive adhesives and permeation enhancers,including their type and concentration.Subsequently,the ginger oil patches were optimized and subjected to content determination and property evaluations.A PTSD mouse model was established using the foot-shock method.The therapeutic effect of ginger oil patches on PTSD was assessed through pathological sections,behavioral tests,and the evaluation of biomarkers such as tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),brain-derived neurotrophic factor(BDNF),and melatonin(MT).Results: The results demonstrated that ginger oil patches exerted therapeutic effects against PTSD by inhibiting inflammatory responses and modulating MT and BDNF levels.Pharmacokinetic experiments revealed that ginger oil patches maintained a stable blood drug concentration for at least one day,addressing the rapid metabolism drawback of 6-gingerol and enhancing its therapeutic efficacy.Conclusions: Ginger oil can be prepared as a transdermal drug patch that meets these requirements,and the bioavailability of the prepared patch is better than that of oral administration.It can improve PTSD with good patient compliance and ease of administration.Therefore,it is a promising therapeutic formulation for the treatment of PTSD.

Unveiling the adaptation strategies of woody plants in remnant forest patches to spatiotemporal urban expansion through leaf trait networks

Background:With the expansion of urban areas,the remnants of forested areas play a crucial role in preserving biodiversity in urban environments.This study aimed to explore the impact of spatiotemporal urban expansion on the networks of leaf traits in woody plants within remnant forest patches,thereby enhancing our understanding of plant adaptive strategies and contributing to the conservation of urban biodiversity.Methods:Our study examined woody plants within 120 sample plots across 15 remnant forest patches in Guiyang,China.We constructed leaf trait networks (LTNs) based on 26 anatomical,structural,and compositional leaf traits and assessed the effects of the spatiotemporal dynamics of urban expansion on these LTNs.Results and conclusions:Our results indicate that shrubs within these patches have greater average path lengths and diameters than trees.With increasing urban expansion intensity,we observed a rise in the edge density of the LTN-shrubs.Additionally,modularity within the networks of shrubs decreased as road density and urban expansion intensity increased,and increases in the average path length and average clustering coefficient for shrubs were observed with a rise in the composite terrain complexity index.Notably,patches subjected to‘leapfrog’expansion exhibited greater average patch length and diameter than those experiencing edge growth.Stomatal traits were found to have high degree centrality within these networks,signifying their substantial contribution to multiple functions.In urban remnant forests,shrubs bolster their resilience to variable environmental pressures by augmenting the complexity of their leaf trait networks.

Deep Global Multiple-Scale and Local Patches Attention Dual-Branch Network for Pose-Invariant Facial Expression Recognition

Pose-invariant facial expression recognition(FER)is an active but challenging research topic in computer vision.Especially with the involvement of diverse observation angles,FER makes the training parameter models inconsistent from one view to another.This study develops a deep global multiple-scale and local patches attention(GMS-LPA)dual-branch network for pose-invariant FER to weaken the influence of pose variation and selfocclusion on recognition accuracy.In this research,the designed GMS-LPA network contains four main parts,i.e.,the feature extraction module,the global multiple-scale(GMS)module,the local patches attention(LPA)module,and the model-level fusion model.The feature extraction module is designed to extract and normalize texture information to the same size.The GMS model can extract deep global features with different receptive fields,releasing the sensitivity of deeper convolution layers to pose-variant and self-occlusion.The LPA module is built to force the network to focus on local salient features,which can lower the effect of pose variation and self-occlusion on recognition results.Subsequently,the extracted features are fused with a model-level strategy to improve recognition accuracy.Extensive experimentswere conducted on four public databases,and the recognition results demonstrated the feasibility and validity of the proposed methods.

Observing eddy dye patches induced by shear instabilities in the surf zone on a plane beach

The effects of surf zone eddy generated by alongshore currents on the deformation and transport of dye are still poorly understood,and related tracer release experiments are lacking.Therefore,a tracer release laboratory experiment was conducted under monochromatic,unidirectional incident waves with a large incident angle(30°)on a plane beach with a 1:100 slope in a large wave basin.A charge-coupled device suspended above the basin recorded the dye patch image.The evolution of eddy dye patch was observed and the transport and diffusion were analyzed based on the collected images.Subsequently,a linear instability numerical model was adopted to calculate the perturbation velocity field at the initial stage.The observation and image processing results show that surf zone eddy patches occurred and were separated from the original dye patches.Our numerical analysis results demonstrate that the structure of the perturbation velocity field is consistent with the experimental observations,and that the ejection of eddy patches shoreward or offshore may be ascribed to the double vortex.