Aim: To systematically evaluate prevalence and clinical characteristics of adverse effects of antidepressants and OTC drugs interactions in a retrospective chart review.Methodology: Dataset of 1,145 registered adverse events were evaluated. Reports were selected for further analysis if pharmacoepidemiological avaluation indicated the presence of high probability of a causal relationship between antidepressants and OTC interaction and the occurrence of side effect. Following variables were extracted from the records: sex, age, medical comorbidities, antidepressant and other concomitant medications, clinical consequences ant the possible interaction mechanisms.Results: 368 showed causal relationship with the simultaneous use of antidepressant with another drug. 15 adverse events (4%) were related to the use of OTC medicine, particularly omeprazole, diphenhydramine, Japanese ginkgo biloba, ibuprofen, diclofenac and sildenafil. All of the analysed side effects were categorized as the result of pharmacokinetic interactions. Here we report identified OTC drugs with corresponding antidepressants and clinical manifestations of DDI. Omeprazole: agomelatine , fluoxetine , sertraline , escitalopram . Diphenhydramine: sertraline , paroxetine , duloxetine . Japanese ginkgo biloba: citalopram (bradycardia), trazodone , mianserine (restless legs syndrome). Diclofenac: escitalopram , and fluoxetine (restless legs syndrome). Ibuprofen: agomelatine (anxiety and nausea), sertraline and omeprazole (QTc prolongation). Sildenafil: fluoxetine  and sertraline .Conclusion: The use of OTC drugs by the patients should be monitored. Pharmacokinetic interactions between nonprescribed medicines and antidepressants may increase concentration and severity of side effects of latter ones. The use of over-the-counter (OTC) drugs is an ubiquitous phenomena, and the number of patients undertaking non-prescribed medication is increasing. Studies show that the rate of those practices in developing countries may reach up to 90% . ImportaThe group of patients particularly vulnerable to the occurrence of side events are those receiving psychopharmacological treatment . PsychotPolytherapy, defined as the use of two or more drugs at the same time, is a common phenomenon in clinical psychiatry. In the USA, up to one third of the patients received at least three psychotropic drugs and over the time this proportion is rising . SimultaGiven the popularity of both antidepressants and OTC drugs, DDI between those two groups of medicines should be a common phenomenon. However, this topic has not been extensively researched. The aim of our study is to evaluate the incidence and characteristics of adverse interactions of antidepressants and OTC drugs in a retrospective chart review.A retrospective chart review was performed to evaluate the prevalence and clinical characteristics of DDI of antidepressants and OTC drugs. Analysis was performed by all the authors. The dataset involved reports on the occurrence of adverse reactions being the consequence of adverse interactions between simultaneously used drugs. They were evaluated at the University Center for Monitoring and Research on Adverse Drug Effects, Department of Clinical Pharmacology at Jagiellonian University Medical College in Cracow. It is one of the Regional Centers that, in accordance with the legal acts in Poland, formally monitor and report complications related to the pharmacotherapy. Additionally, this organisation provides specialist consultations for both hospitals and clinics from the Lesser Poland, Silesian, Holy Cross and Subcarpathian regions. The Center collaborates closely with the Department of Affective Disorders of the Jagiellonian University Medical College due to the significant increase of the side events associated with the use of psychotropic drugs. Annually, the Center makes approximately 850–1,100 consultations.In this study we have analysed reports that came from all over the Poland in the period between January 2017 till March 2018. They were selected for further analysis when the following criteria have been met: 1) patient received at least one antidepressant drug, 2) patient used at least one OTC medicine, 3) the presence of a high probability of a causal relationship in terms of pharmacokinetic, pharmacodynamic interactions or the interactions associated with aggregation of side effects associated with the simultaneous use of antidepressant and OTC drugs indicated by pharmacoepidemiological analysis. Data extracted from 15 adverse events presenting a causal relationship with the use of antidepressants and OTC drugs is presented in In this study we have performed the first retrospective chart review of the adverse events caused by the simultaneous use of antidepressants and OTC drugs, based on a thorough analysis of the 1,145 reports. 4% of the adverse events caused by interaction of antidepressants with other drugs, were caused by their simultaneous use with OTC medication, particularly omeprazole, diphenhydramine, Japanese ginkgo biloba, ibuprofen, diclofenac and sildenafil. In this restrospective chart-review, as in our previous studies , the meaOmeprazole was the most commonly used OTC drug associated with adverse events evaluated in our study. Proposed mechanisms of those interactions involve interaction with cytochrome 450 isoenzymes responsible for antidepressants metabolism. Particularly, through inhibition of CYP2C9 and CYPC19 omeprazole increases the concentration and side effects of agomelatine , sertraline (vertigo and yawning), fluoxetine  . In two Diphenhydramine is a first-generation antihistamine drug that acts as an inverse agonist on the H1 receptor with a serotonin reuptake inhibitor property , which wFlavones and flavonols contained in the raw Japanese ginkgo biloba material may be potentially used by patients as a self-management aimed to improve brain blood supply, mental performance, memory and to decrease severity of depressive symptoms . Drug inMost of the studies evaluating interactions between antidepressant drugs and NSAIDs are focused on SSRI. Main groups of those interactions involve: 1) inhibition of platelet aggregation and function through the different mechanisms; 2) independent effect of SSRIs without direct pharmacokinetic interaction, e.g. increase of gastric acid secretion; 3) pharmacokinetic interaction through the inhibition of the CYP2C9, leading to the increased concentration of antidepressants metabolised by this isoenzyme . Most ofErectile impotence is a well-documented, common symptom of depression as well as a side effect of SSRI . SildenaThe most common group of antidepressants associated with the occurrence of adverse events in our retrospective chart review were SSRIs. All of the DDI described in our study represented pharmacokinetic mechanisms related to the inhibition of cytochrome P450 and the increase of those drugs concentrations and side effects. However, it should be noted that the use of SRRIs may associated with occurrence o the serious adverse effect in form serotonin syndrome. Many OTC drugs, such as dextromethorphan, purple echinacea, ginseng or ginkgo biloba  may incrIn this restrospective chart-review, as in our previous studies , the mea- The pharmacokinetic profile of the patients medications should be investigated in order to evaluate whether there is overlap between cytochrome P450 isoenzymes involved in the metabolism of the drugs used, what may affect their concentration.- OTC drugs can interact with each other, which may cumulatively increase the concentration and side effects of antidepressants.- Particular attention should be paid in situations where an antidepressant is used in the maximum dose or the dose is titrated rapidly, because pharmacokinetic interaction with OTC drug may lead to exceeding the therapeutic concentration.- Interactions of antidepressants and OTC drugs may result of life-threatening adverse events, e.g. myocardial infarction described in our study.- Patients should be asked by doctors (primary care physician or psychiatrist) as well as the pharmacist about the usage of OTC drugs and informed about possible side effects caused by their simultaneous use with antidepressants.- The use of OTC drugs by the patient should be described in the medical records in order to be able to monitor the adverse events associated with the use of these drugs.


This paper aims at proposing an augmented sensing method for estimating volumetric water content (VWC) in soil for Internet of Underground Things (IoUT) applications. The system exploits an IoUT sensor node embedding a low-cost, low-precision soil moisture sensor and a long-range wide-area network (LoRaWAN) transceiver sending relative measurements within LoRaWAN packets. The VWC estimation is achieved by means of machine learning (ML) algorithms combining the readings provided by the soil moisture sensor with the received signal strength indicator (RSSI) values measured at the LoRaWAN gateway side during broadcasting. A dataset containing such measurements was especially collected in the laboratory by burying the IoUT sensor node within a plastic case filled with sand, while several VWCs were artificially created by progressively adding water. The adopted ML algorithms are trained and tested using three different techniques for estimating VWC. Firstly, the low-cost, low-precision soil moisture sensor is calibrated by resorting to an ML model exploiting only its raw readings to estimate VWC. Secondly, a virtual VWC sensor is shown, where no real sensor readings are used because only LoRaWAN RSSIs are exploited. Lastly, an augmented VWC sensing method relying on the combination of RSSIs and soil moisture sensor readings is presented. The findings of this paper demonstrate that the augmented sensor outperforms both the virtual sensor and the calibrated real soil moisture sensor. The latter provides a root mean square error (RMSE) of  The measurement of soil moisture is a key task in a large number of applications in the context of precision agriculture. In fact, autonomous crop watering is included in most precision-agriculture systems, where information concerning soil moisture is one of the main drivers for the activation of irrigation systems. Despite the wide availability of sensors in the market, an accurate measurement or estimation of volumetric water content (VWC) in soil is still a challenging task which requires expensive devices. Indeed, low-cost sensors can easily be found, but they are, in practice, so crude that they are mainly used to distinguish between dry and wet soils. While the latter information may suffice for watering plants in small gardens, it is not sufficiently accurate for precision-agriculture applications, where water usage needs to be optimised to improve overall system efficiency. Indeed, intensive crops may span over thousands of square kilometres, thus requiring pervasive wireless underground sensor networks (WUSN) for remote monitoring ,2,3,4. IThe big picture of the IoT depicts low-power wide-area network (LPWAN) technologies as the ones garnering the most attention. Due to their long coverage, up to tens of kilometers, and their limited power consumption, they naturally suit harsh contexts, such as underground. Similarly, their usage is especially fostered by their remarkable robustness, in particular for those operating in sub-GHz bands, . Among them, long-range (LoRa) modulation is the one that has undergone the greatest spread. This is due to its exceptional features, the transceivers low cost and high availability on the market naturally eases the deployment of network infrastructures. Thanks to the associated low-power wide-area network (LoRaWAN) MAC-layer protocol, they are capable of handling up to thousands of devices, enabling the set up of pervasive monitoring facilities. In light of this, LoRa-based systems have been extensively adopted in the most diverse application scenarios. Smart City  or SmartUnderground data transmission is especially challenging since the modelling of the quality of the radio channel can be affected by a wide range of different parameters, including the soil composition and soil moisture. In particular, a relation emerged between the level of VWC and radio parameters, such as the received signal strength indicator (RSSI), especially in LoRaWAN UG2AG links . Such reProposal of a set of ML-based methodologies for the estimation of VWC exploiting a WUSN enabled by the LoRaWAN protocol;Presentation of a threefold sensing technique for VWC, which, respectively, are the calibration of a real cheap soil moisture sensor (that is not suitable for measuring VWC) exploiting its raw readings, the realisation of a fully virtual sensor estimating VWC from LoRaWAN RSSIs, and the implementation of an augmented sensor for VWC, taking as input both the soil moisture sensor raw readings and the LoRaWAN RSSIs;Introduction of a filtering process to enhance the performances of the aforementioned sensing techniques.This paper exploits the preliminary results of a previous work , which sThe rest of the paper is organised as follows: The LoRa modulation and LoRaWAN protocol are widely adopted amongst a plethora of applications due to their robustness, reliability and wide operating rage. Industrial monitoring and “smart cities” are common application contexts, and harsh environments, such as marine  or underIt is well-known that propagation in radio communication systems depends on characteristics of the medium. For instance,  analysesDespite the clear influence of soil moisture on radio transmission performances, a surprisingly limited number of investigations have sought to exploit this link to infer VWC values from measurements related to radio parameters. In , the estML techniques have been extensively applied to the prediction of soil moisture levels. However, most of the works in the literature exploit different typologies of datasets to that of the approach presented in this paper. A large number of contributions have focused on the exploitation of remotely sensed data; two comprehensive reviews can be found in ,31. The The majority of the works in remote sensing address architectures, communication schemes, protocols, or sensing technologies, while a limited number focus on the actual usage of the collected data. In , soil moThe concept of path loss is useful to understand the relationships between soil moisture and radio parameters. Relevant literature contributions on underground path loss are found in ,44,45,46Path loss can be found from the modified Friis equation:PRX=PTX+Gf is the carrier frequency expressed in MHz. This kind of loss strictly depends on the covered distance and the carrier frequency. In particular, a logarithmic dependency arises, as can be seen in The aboveground losses Refraction losses occurring at the soil-air interface are computed according torend see c.m, while f is the carrier frequency, g use of ,48:(5)LUents see , while tents see . Figure  (MBSDM) ,50 and t (MBSDM) .The MBSDM can be adopted in the range 0.045–26.5GHz and requires the VWC, the carrier frequency and the percentage of clay related to soil composition as inputs. Conversely, the ITU model for path-loss estimation due to soil implies knowledge of the carrier frequency, the temperature, the percentages of either sand and clay, the specific gravity (SG) , the VWC and the bulk density (BD) of the soil samples.Regardless of the model employed in the evaluation of the real and imaginary parts of the effective soil permittivity, in both cases, VWC is one of the parameters to be evaluated. This suggests the reversibility of this relationship to derive VWC from radio transmission parameters, in particular by resorting to channel losses or received power.In order to gather the dataset, an experimental setup was devised to collect the values from the low-cost moisture sensor, as well as the RSSIs related to the transmitted LoRaWAN packets enclosing the sensor measurements in their payloads. Tests were accomplished in the laboratory, pursuing the aim of controlling the trials and limiting the number of involved variables .The dataset was acquired by making use of a plastic case that was filled with sand, while the sensor node was buried within it, thus becoming an IoUT node. The plastic case was a  of 2.66 .The IoUT sensor node  band, and by making use of the minimum spreading factor (SF) provided by the LoRaWAN protocol  and a bandwidth of part see  so as toThe adopted gateway was an LG308 manufactured by Dragino. Its sensitivity depends on the SF and the exploited bandwidth of the transmitted packets. Therefore, for the aforementioned transmission test parameters, the gateway had a sensitivity of ribed in , since tThe adopted methodology worked as shown in the flowchart presented in The collected dataset during the tests included 27,000 data points. The resulting dataset was exploited to implement several solutions based on ML techniques, aiming at estimating VWC values combining the different typologies of data. First of all, the data was organised in a table, containing the measurements of the low-cost sensor , the RSSI measurements provided by the LoRaWAN gateway, and the reference values for VWC to be exploited as labels. In other words, the dataset labels belong to the set The moisture sensor raw readings are very roughly related to reference VWCs due to the coarse precision of the sensor. From Therefore, simple VWC estimates would be coarse if they only rely on sensor raw readings, or almost unfeasible if they are grounded on the reversibility of the theoretical model taking as input the RSSIs. That is the reason why ML models are exploited. This understanding is also confirmed by Raw readings only;RSSIs only;Raw readings together with RSSIs.Different ML-based regressors are tested, including linear techniques, polynomial-augmented penalised linear techniques, and ensemble methods, including forest tree and gradient-based tree regressors. For each model, the same targets  are chosen, while different sets of variables are used for the training:In so doing, each of the ML models has three versions for the functions of the exploited training data. In particular, the first version only processes raw readings in order to map them into VWC estimates to obtain a calibrated version of the real soil moisture sensor, the second one describes a relationship between RSSIs and VWCs aiming at setting up a virtual sensor for VWCs, while the third estimates VWCs simultaneously accounting for both soil moisture sensor raw readings and RSSIs, resulting in a full-fledged augmented sensor for measuring VWC. The considered ML models are the following: ordinary least squares (OLS) regressor, ridge regressor, least absolute shrinkage and selection operator (LASSO) regressor, logistic regressor, support vector machine (SVM) regressor, random forest (RF) regressor, gradient boost (GB) regressor and extreme gradient boosting (XGBoost) regressor. Such models are taken into account because they are the most adopted for regression purposes.Data was divided into training and test sets  keeping an equal balance between different VWC reference values in the training and test sets. Moreover, in order to improve performances, a hyperparameter optimisation procedure was applied by resorting to a k-fold cross-validation approach with five non-overlapping folds. Indeed, the performances of each ML model depend on its hyperparameters. For instance, in the case of ensemble tree methods, some of them are the number of trees and their maximum depth, or the learning rate that shrinks the contribution of each tree. For each model, the values of relative hyperparameters were chosen by dividing the training set into training and validation subsets and exploring performances from values of the hyperparameters spanning in a grid . Finally, the root mean squared error (RMSE) was exploited as a loss function during the training phase.The consistency between the training and test procedures, as well as the absence of overfitting during training, arises by comparing the values of Generally, the ensemble tree methods ensured satisfactory results . Indeed,Histograms of regression errors for each of the three model versions are provided in The analysis of possible relationships between the estimation errors and VWCs for each of the model versions is given in The virtual sensor exploiting only the RSSIs is able to provide rough VWC estimates because its RMSE is In order to reduce the outliers see , and conConsequentially, with regard to the augmented sensor , filtering enhances the estimation error probability see , and redFinally, factors such as burial depth, soil composition or As stated in The main limitation of the proposed approach is common to any data-driven method. As the proposed methodologies are grounded in data collection strictly dependent on the soil topology, they necessarily require a starting stage during which they have to undergo a setup and tuning step, implying the collection of additional data related to the soil at hand. Nonetheless, all of the proposed techniques  can be replicated for whichever soil composition, highlighting the methodology adaptability.The aim of this paper was to propose the usage of ML-based techniques to improve the accuracy of LoRaWAN-based wireless soil-moisture-sensing systems, aiming at measuring VWC. In particular, the relation among VWC and RSSIs was exploited in two ways: firstly, by setting up a virtual sensor obtaining a rough VWC estimation by only exploiting RSSIs; secondly, by combining these values with measurements acquired with a low-cost, low-precision soil moisture sensor, thus achieving an augmented sensor. The proposed method was assessed by means of a measurement campaign aimed at collecting a real dataset containing both RSSIs related to UG2AG LoRa links, and the low-cost, coarse soil moisture sensor readings as a function of several levels of VWC. Such tests were devised in a controlled experimental environment by resorting to a plastic case filled with sand. It was gradually watered to augment the VWC, while LoRaWAN RSSIs and the raw sensor readings were regularly acquired. In so doing, nine levels of VWC were obtained, during each of which three transmission batches of 1000 LoRaWAN packets were executed.Acquired data was used to evaluate the performances of different ML regression algorithms for the estimation of VWCs using three different sets of variables, leading to the identification of an ensemble-tree-based algorithm as the best model for the specific target . In the first case, only raw readings coming from the coarse sensors are exploited, leading to a model implementing the calibrated version of the real sensor. Then, only RSSIs are used, thus obtaining a virtual sensor. Finally, both RSSIs and raw readings from the soil moisture sensors are employed, turning the relative model into a full-fledged augmented sensor. The results demonstrated that the virtual sensor is able to roughly estimate the VWC level, the calibrated real sensor gives intermediate performances, while the augmented sensor provides the best VWC estimates. Moreover, the augmented sensor can be further improved by applying a post-processing filtering technique , ensuring an estimation error which is comparable to the adopted quantization step for VWCs during the measurement campaign for the dataset gathering.The results of this paper can be of significant importance for the actual deployment of distributed measurement systems in application scenarios which are typical of precision agriculture contexts. Indeed, a large number of connected monitoring devices is generally required to cover the large surfaces of cultivation and crops. The availability of low-cost devices and accurate methods may boost the adoption of intelligent systems, with notable benefits for overall reduction in the environmental impact of industrial crops. Nonetheless, this paper’s findings can be easily applied wherever WUSN are concerned.The current limitations of the proposed methodology are those that any data-driven approach implicitly entails. Indeed, because the proposed sensing techniques are based on a dataset gathering information related to a single typology of soil , extending them to any soil composition may require an initial setup and tuning stage needing the collection of additional data related to the soil at hand. However, this also means that the presented methodology can be fully repeated for whichever soil composition is considered. Nonetheless, in future work, provided there is accurate soil analysis providing its composition, the proposed methodology can be easily improved by injecting in the models the knowledge coming from the theoretical analysis explained in


Diamond wire sawing is the main machining technology for slicing various brittle materials, such as crystalline silicon, SiC, and NdFeB. Due to their high hardness and high brittleness, as well as the ease with which the surfaces of machined materials are damaged, it is difficult to further improve the sawing efficiency and the surface quality based on research conducted on the original machining method. In this paper, a vibration-assisted diamond wire sawing method is proposed. We analyzed the impact of load on the ingot, motion trajectory, and sawing depth of the abrasive particles, and a macroscopic sawing force prediction model for the vibration-assisted sawing method was established and verified via experiments. Based on the single-wire-sawing experiment and prediction model, the influences of the vibration parameters and sawing parameters on the sawing force were determined. The influences of vibration assistance on the surface quality, including the roughness profile, waviness profile, thickness profile, Ra, and Rz, were explored through single-wire-sawing experiments, and the influences of vibration assistance on the geometric parameters of slices, such as the total thickness variation (TTV) and warp, were explored through multi-wire-sawing experiments. It was found that vibration-assisted sawing can reduce sawing force and improve surface quality. In recent years with the development of material science and industrial technologies, brittle materials, such as sapphire, NdFeB, and SiC, have been widely used in the electronic industry, photovoltaic industry, aerospace industry, and other fields. Fixed abrasive wire sawing has become the main machining method for sawing brittle materials due to its technical advantages, such as narrow incision, good surface quality, and strong sawing ability, which help it to achieve low total thickness variation (TTV), a smaller damage layer, and fewer surface cracks ,2,3.In the case of diamond wire saws, the existing processing efficiency and wafer quality are difficult to further improve by changing the process parameters. Therefore, some scholars have tried to optimize diamond wire sawing technology using vibration-assisted methods.For conventional diamond wire sawing, scholars have explored the influence of the sawing parameters on the sawing force through experiments and have established corresponding mathematical models, which suggest that increasing the size of the abrasive particles, increasing the wire speed, and reducing the feed speed ,5,6 may In the process of establishing a cutting force model, it is necessary to analyze the mechanisms of crack damage and material removal between abrasive particles and the material surface. Important parameters, such as the force on abrasive grains, cutting depth, and undeformed chip thickness, are determined during the material removal process. Shengyao Yang  revealedSince it is difficult to further improve sawing efficiency and surface quality and there is inevitable surface or subsurface damage, some studies have tried to apply ultrasonic vibration-assisted technology to diamond wire sawing. Liaoyuan Zhang  tried toIt can be seen from the above research that applying vibration assistance can reduce the sawing force and improve both the sawing efficiency and the surface quality of slices. However, diamond wire sawing has gradually transformed from single-wire sawing to multi-wire sawing. The wire mesh formed via multi-wire sawing has an important effect on sawing quality . The oriBased on the above research, vibration assistance is applied to a working table carrying ingots in the direction of the wire speed. By analyzing the kinematic characteristics, the impact load, and the sawing depth of all the abrasive particles on the wire, an equivalent macroscopic sawing force model is established. A vibration-assisted single-wire-sawing experiment is used to verify the sawing force model and to explore the influences of the sawing parameters and vibration parameters on the sawing force. The influences of vibration assistance on the roughness profile, waviness profile, Ra, and Rz are explored through vibration-assisted single-wire sawing. The influences of vibration assistance on TTV and warp are explored through vibration-assisted multi-wire sawing.The wire mesh of a multi-wire sawing machine was composed of a diamond wire wound sequentially on grooved wheels, and its cross-section was taken to analyze vibration-assisted diamond wire sawing .Taking the wire speed on the right as an example, As shown in The sawing forces generated by the sawing ingot are mainly composed of the chip deformation force and friction force. By analyzing the theoretical principle of chip deformation and the tribological theory, the sawing forces termined :(1)Fen=K2).In the formula,  follows :(2)Fn=2∫In the formula, As is shown in When analyzing the vibration impact, the maximum sawing depth In the formula, According to the principle of equal volume, the equivalent action time of a single abrasive particle in one vibration period According to the impulse theorem, the average normal sawing force and the average tangential sawing force of a single abrasive particle in one vibration period can be expressed as follows:With reference to Formula (2) for sawing forces in conventional sawing, the following formula determines the sawing forces in vibration-assisted wire sawing:As shown in The vibration-assisted platform consisted of three parts: the servo motor, the eccentric device, and the working table. The eccentric device converted the rotation of the servo motor to the vibration of the working table along the wire speed. The motion of the vibration platform and ingot could be monitored by the accelerometer. Through the control system of the vibration-assisted machine tool, the selection and adjustment of vibration parameters and sawing parameters could be realized.The ranges of the vibration parameters, sawing parameters, and ingot size in the experiment equipment are shown in To verify the vibration equivalent sawing force model, it was necessary to obtain the We used the same batch of HT250, stainless steel, NdFeB, and SiC as experimental ingots after aging treatment, and the ingots were square. In the single-wire-sawing experiment, the surface roughness and surface profiles were scanned and measured using a Bruker Dektak XT instrument to obtain the Ra, Rz, roughness profile, waviness profile, and thickness profile. In the multi-wire-sawing experiment, the TTV and warp of the slices were measured to evaluate the average surface quality.According to the adjustable value range of the experimental equipment, the same batch of HT250 was selected to ensure that the material properties were essentially the same. The orthogonal method was used to explore the influences of vibration frequency, feed speed, wire speed, and tension on the tangential sawing force and normal sawing force in vibration-assisted single-wire sawing. To verify the accuracy of the vibration equivalent sawing force model, we selected certain vibration parameters and sawing parameters .The experiments were carried out according to the method described in Through the description of the above experimental results, it was concluded that vibration assistance could effectively reduce the sawing forces during sawing and that vibration frequency was one of the important parameters affecting the sawing force.It was predicted that, with an increase in It was predicted that, with an increase in It was predicted that, with an increase in An orthogonal experiment method was adopted, and the following discussion results were arrived at by comparing the experimental results with the theoretical results:The relative error between the experimental values and the theoretical values was relatively small, which verified the accuracy of the sawing force prediction model;With increase in the vibration frequency (0–50 Hz), the sawing forces Sawing parameters, including wire speed, feed speed, and wire tension, affected the impact load between the abrasive particle and the ingot and further influenced the optimization effect of vibration assistance on the sawing forces HT250, NdFeB, stainless steel, and other materials were selected to explore the influence of vibration assistance on the roughness profiles, Ra and Rz values, waviness profiles, and thickness profiles of the slices. The change trend in the waviness profile was consistent with that in the thickness profile, which reflected the quality difference on the same surface. Ra and Rz at different positions on the same surface were scanned and measured, and the average values and standard deviations of Ra and Rz were obtained. The selected vibration parameters and sawing parameters are shown in The experiments were carried out according to the method described in The abrasive particles produced a polishing effect on the surfaces during vibration-assisted sawing;Vibration assistance could reduce the surface roughness values of Ra and Rz for different materials;The worse the stability of the surface quality in conventional sawing, the more significant the optimization effect of vibration assistance on quality difference.Based on the comparative experimental results, the following discussion results could be drawn:3 of TTV and warp were obtained. The vibration parameters and sawing parameters are shown in The single-wire-sawing experiment was extended to a multi-wire-sawing experiment, and the geometric parameters of slice TTV and warp values with different vibration parameters and sawing parameters were measured to explore the influence of vibration assistance on average surface quality. The average values, maximum values, and standard deviations σAccording to the experimental method shown in In In TTV and warp are important standards for measuring the quality of slices. In order to meet the needs of large-sized, high-precision, multi-slice processing, it is necessary to obtain good surface precision with good TTV and warp values. An unqualified TTV affects the chip pass rate and the production process, and the evaluation standards also put forward strict requirements for chip quality. Based on the above results, we concluded that the average values, maximum values, and standard deviations of TTV and warp were relatively reduced and that the sawing stability and surface quality were improved when using multi-wire sawing. Vibration-assisted technology could improve the pass rate of slices and optimize the production process.In this paper, a method of applying vibration assistance to ingot slicing was presented that attempted to extend vibration assistance to the frequency range of 0–50 Hz. A sawing force prediction model for vibration-assisted wire sawing was established. The prediction model described the theoretical relationship between the process parameters and sawing forces. The relationships between sawing forces and process parameters, such as vibration frequency, wire speed, feed speed, and wire tension, were explored through experiments. The effects of vibration assistance on surface quality were explored in both single-wire-sawing and multi-wire-sawing contexts.In wire sawing, the deformation of the wire saw and the vibration assistance affected each other;During vibration-assisted wire sawing, the sawing forces could be reduced by different sawing parameters;Vibration frequency, feed speed, wire speed, and wire tension were important factors that affected the sawing force, and the effect of vibration assistance on the sawing force was comprehensively affected by the sawing parameters and vibration parameters;The surface roughness, quality difference on the same surface, TTV, and warp were all reduced during vibration-assisted wire sawing;It was predicted that the polishing effect of abrasive particles on the surface would be enhanced and that chip removal would be improved when using vibration-assisted sawing;The selection and matching of materials, vibration parameters, and sawing parameters were based on the prediction force model and the experiments described in this paper.The conclusions that could be drawn based on the results were as follows:


The aim of this Special Issue is to highlight the close functional and highly conserved link between innate immunity, homeostasis maintenance, inflammation, tissue remodeling and regeneration. In the last few decades, the rules on the use of vertebrates in biological trials have become increasingly stringent and, in this context, invertebrates are used as complementary animal models, considering that they are cost-effective and easy to handle. Here, original articles and reviews on cells and mechanisms involved in immunity, in the regulation of inflammation and tissue regeneration processes, focused on invertebrate models, are gathered.Considerable efforts have been made to shed light on phylogenetic distribution and developmental patterns of wound healing and regeneration mechanisms, as well as on their ecological value. Comparative research on regeneration and its relationship to the immune system has been a long-standing concern. Most likely, the capability to regenerate whole organs and body parts concurs with the origin of multicellularity and many invertebrate models have relevant regeneration abilities, unlike in vertebrates, in which this skill is significantly reduced.In the paradigm of acquired and adaptive immunity, anamnestic memory responses are the central feature of the adaptive immune system that evolved in our vertebrate ancestors, together with cicatrisation following tissue injury, which is disadvantageous to full regeneration.Anemonia viridis, focusing precisely on the rearrangement of the extracellular matrix in the mesoglea of this Mediterranean benthic species. They analysed the event from a biochemical, biometric, morphological and histological perspective, in unamputated and injured polyps. In particular, modifications in mesoglea stiffness were observed during tentacle regrowth, the production of metalloproteases probably attributable to fibroblast-like cells, differentiated from amebocytes, and which could play a role in the reshaping of the ECM and the deposit and spatial arrangement of collagen I fibrils. Indeed, it might act as a scaffold and as a guide for the reconstruction of missing parts. Moreover, the regrowth of the oral tentacles was followed for 14 days at the macroscopic level. The wound closed after 24 h from the lesion, appeared less pigmented, due to the absence of the endosymbiont of Symbiodinium genus, lost from the tissues after injury. The sprawling regrowth was already noticeable after 7 days and new pigmented 5 mm sized tentacles were visible after 14 days from amputation. At 14 days from cut, the repigmentation was correlated to the repopulation of the tissues by the zooxanthellae, which also appear to play an important role in elongating the regenerating tentacles. In this contest, the activity of alkaline phosphatase (ALP), as a typical biochemical marker associated with regenerative events and related to nematocyte and epithelial cell differentiation in cnidarian, was quantified. The researchers showed an increase in the ALP activity in tentacle extracts in all three experimental time points. On the contrary, the enzyme amount remained at a constant level in the body extract.In the last few years, the key role of the extracellular matrix (ECM) during several physiological processes has become evident, including in regeneration scenarios. Parisi et al., 2021 [A. viridis tentacle regeneration, mesoglea move from a laxist network to an organised one of collagen fibril bundles. In addition, immunocalization tests showed that the visco-elastic behaviour of the mesoglea from the neosynthesis of collagen type I from hyaline amebocytes bFGFR+ act as fibroblast-like cells and are involved in collagen structure arrangement.Histological analyses showed, for the first time, that in Eisenia andrei. They focused on, with a new point of view, its rostral and caudal regeneration after injury. The classic steps of annelid regeneration in E. andrei with the instantaneous closure of the wound and the completed recovery of lost tissues in four weeks were observed. Moreover, several cellular and molecular modifications in regenerating earthworms were found. Immediately after the wound closure, proliferating cells create the blastema, which appeared clearly at 4 days from injury, characterized by distinct cell types. Proliferation occurs until two weeks in regenerating earthworms.The close link between innate immune response and wound-induced regeneration was also investigated by Bodó et al., 2021 [At the same time, apoptosis occurred during the whole regenerative event, particularly intensive during the wound healing, and a cellular breakdown was detected within four weeks of regenerating anterior and posterior blastema. Visualization of smooth muscle α (αSMA) by immunofluorescence, only in intact original segments, was strongly visible and so expressed after four weeks of regeneration in the muscle layers, both in anterior and posterior blastema. These remarks appear to rise the conserved role in evolution of αSMA, given its constant de-novo expression/activation in new segments.Histochemical methods evidenced a decrease in pattern recognition receptors and antimicrobial peptides and acid and alkaline phosphatase showed significant alterations between intact and regenerating organisms. The regeneration event appeared clear after two weeks, with an increase in cell proliferation and a reorganization of new actin fibers. Concurrently the apoptosis present during the whole process decreased in two-week regenerating organisms and high-programmed cell death in early blastema formation and in the trans-differentiation phase.Earthworm coelomocytes were involved in segment regeneration in combination with the production of several antimicrobial peptides (AMP), improving the reparation of damaged axons in central nervous system (CNS). However, authors evidenced how immune-response-related mRNA expression was reduced compared to intact animals, common in other earthworms and in other species during regeneration.E. andrei, investigated the effects of a manganese metalloenzyme arginase activity in coelomocytes, subjected to several and multiple-stressors, including immunostimulant lypopolisaccharide (LPS), hydrogen peroxide (H2O2), metals ions, parasite infection, wound-healing and five days of fasting. Normally, the enzyme is found both in the animal and vegetal kingdom and it plays a key role in ureagenesis, tissue regeneration and in macrophage polarization. An upregulation in the enzyme expression was related to cell growth and collagen synthesis, motility and detoxification from free-radical ions.Moreover, Homa et al., 2021 , in E. aOverall, this study suggested the arginase role in the dual aspect of the post-injury process: immune response involvement and wound healing. Indeed, it was observed that in coelomocytes affected by LPS, hydrogen peroxide and ions up-regulate the arginase activity in in-vitro stimulation, as well as in infected specimens with nematodes or subjected to segment amputation. No remarks were evidenced in fasting organisms for five days; indeed, the number and the composition of coelomocytes did not change. Furthermore, the study confirmed that the coelomocytes metalloenzyme is suppressed by L-norvaline.Mytilus galloprovincialis haemocytes. The authors evidenced the effects on cell motility and the consequent activation of the respiratory burst. Anandamide (AEA) and capsaicin were demonstrated to inhibit the immunity response mediated by the haemocytes, and the selective antagonists CB2 and TRPV1 receptors revert their inhibitory effects. There was a clear morphological shift, from ameboid to rounded shape, in M. galloprovincialis haemocytes after the exposure to AEA and 2-AG. The phagocytosis tests performed exhibited the capability of AEA to control respiratory burst with a diminution in luminescence generation. The suppressive effects of AEA on haemocyte phagocytosis were observed at 10 nM and 100 nM in a dose-dependent manner, and the lowest concentration (1 nM) was inefficient. Thus, it was shown that the haemocytes of the mussel are not only able to synthesize, degrade and transport the major eCBs, but are also able to express the functional eCBs-binding CB2 and TRPV1, but not CB1, receptors and also exposed the suppressive impact on phagocytosis caused by capsaicin.In this Special Issue, Mosca and co-authors  reportedParacentrotus lividus, where the authors deepened the protein family of Transformer protein (Trf) involved in the immune response to pathogens. The Trf family is highly differentiated and shows several Trf sizes and conformations and these were individuated both in cell-free coelomic fluid and in coelomocytes. The authors characterized, through FACS, five sub-populations with PlTrf protein expression on the cellular membrane. The amount of these cells was higher if the organism was exposed to Escherichia coli. No differences in relative abundance were found in immune challenge with LPS or Vibrio penaeicida, a common sea urchin pathogen. They also demonstrated that the phagocytosis activity against E. coli is inhibited, blocking the functionality of PlTrf present on the cell surface with the anti-Trf antibodies. These results suggest the collaboration of humoral and cell-mediated response to bacterial challenge in P. lividus. Lastly, in spite of the differences between gene and protein sequences within this class, their properties and mechanisms of action, as pointed out by the authors, seem to be preserved.Another interesting step forward in the complex field of innate immune system was addressed by Yakovenko et al., 2021 , in theiPomacea canaliculata. They followed three months of the regeneration process for an adult cephalic tentacle, normally used in food searching, co-specific recognition and orientation. After 72 h post amputation, the authors showed the missing parts recognizable and the epithelial, the connective, the muscular and the neuronal tissues were distinguishable. They also showed that haemocytic recruitment occurs in the early stages of regeneration and, to quantify the number of cells present in the regenerated organs, applied an innovative and specific method. Particularly, they applied the protocol for computer-assisted image analysis to tissue sections stained with haematoxylin–eosin of specimens in regeneration. A significant amount of granular haemocytes in proximity to the blastema 12 h post amputation (hpa), followed by a fast visualized decrease of 24, 48 and 72 hpa.Bergamini et al., 2021  providedThe strength of the protocol developed by Bergamini and co-authors aimed at counting cells in the tissue is due to the absence of the use of a specific cellular marker. Therefore, it represents a valid tool for studying the involvement of immune cells in the regenerative process.Clear results, as in each study reviewed here, are fundamental and useful for the study of regeneration. They provide new methods and important biological information that could be exploited in the medical or biotechnology fields, also exploiting the use of new animal models.


Eddy current (EC) testing has become one of the most common techniques for measuring metallic planar structures in various industrial scenarios such as infrastructures, automotive, manufacturing, and chemical engineering. There has been significant progress in measuring the geometry, electromagnetic properties, and defects of metallic planar structures based on electromagnetic principles. In this review, we summarize recent developments in EC computational models, systems, algorithms, and measurement approaches for planar structures. First, the computational models including analytical models, numerical methods, and plate property estimation algorithms are introduced. Subsequently, the impedance measurement system and probes are presented. In plate measurements, sensor signals are sensitive to probe lift-off, and various algorithms for reducing the lift-off effect are reviewed. These approaches can be used for measureing thickness and electromagnetic properties. Furthermore, defect detection for metallic plates is also discussed. Measuring the defect and physical properties  of a planar structure is important in a range of technological applications, for example, coating surface treatments  and qualForward and inverse problems are the primary theoretical issues of EC testing. The forward problem aims to calculate the frequency-dependent measurements for given metallic objects, while the inverse problem determines the physical properties of the object from the frequency-dependent measurement. The forward problem can be solved by analytical and numerical methods. The analytical methods include the Dodd and Deeds model ,14 for sThe implementation of EC testing consists of a data acquisition system and probe which are used to obtain the measurement signal. The customized and commercial impedance analyzers both enable the measurement of mutual inductance between coils and other magnetic field variables. In general, EC probes designed for plate property estimation and defect scanning contain transmitting (Tx) coils and receiving (Rx) elements. According to the required EM field for specific applications, the structure and dimensions of Tx coils can be selected and optimized. Furthermore, coils or magnetic field sensors are usually chosen as the Rx elements, depending on the induced EM field by test pieces. As compared with previous review articles on eddy current testing ,28,29,30The arrangement of the rest of the paper is as follows: In For the single-layered plate model placed near a probe consisting of Tx and Rx coils, as shown in The inductance can be calculated through numerical integration, whereas it is time-consuming. The truncated region eigenfunction expansion (TREE) method, proposed by T. Theodoulidis and E. Kriezis, recasts the expression as sums rather than integrals ,32, i.e.Since the plate term If the sensor structure is known or can be inferred, one can obtain In the theoretical analysis, the Dodd and Deeds model in Equations (1)–(3) calculates the inductance of plates with infinite radius, whereas in practical measurements, the plates normally have finite radiuses.The TREE method indicates that the EC of the finite-size geometry can be evaluated by considering a certain range of spatial frequencies in the analytical model. By analyzing the analytical solution, R. Huang et al. proposed that the spatial frequency e radius ,34,35, aThen, based on the TREE method, the finite region eigenfunction expansion (FREE) method is proposed which confines the terms of series relating to the plate radius, i.e.:The finite element method (FEM) is a universal computation tool for arbitrary sensor setup and the geometry of a target object. With the support of Galerkin equations , the unkAs compared with analytical methods, the FEM method can adapt to any geometry with high accuracy. However, it has a high computation load and requires a large amount of time to obtain the final solution for a single simulation. An equivalent phenomenon was observed for a thin structure by using an analytical method. From the principle of a plane wave reflected and transmitted at the surface of a nonmagnetic plate, with a geometry of thin thickness, the inductance ified as :(13)ΔL=2Then, considering two plates in different materials and thicknesses, the induced inductance is approximately equal when:The above relationship reduces the computational burden when modeling thin structure samples. Researchers have been devoted to speed up the computation speed to satisfy users’ requirements by using some approximations/techniques, i.e., zero-thickness defect , dedicatThe proposed preconditioning method optimizes the initial guess for solving the system algebraic equations. Combined with the perturbed matrix inversion (PMI) method, a fast FEM approach has been proposed which can effectively shrink the time needed for the crack computation due to the small perturbation . FurtherThe probe tilt is identified as one of the major sources of noise in EC surface inspections . The FEMFollowing studies ,48, applAs the 2D Fourier transform can characterize the rotation and translation function, for the probe shown in Substituting the source terms in Equation (15) by ar coils .For the measurement of a plate with an infinitely long crack, according to the thin-skin model in high-frequency regime ,50, the fined in ,51.The measurements of probe impedance scanning through the finite-size test pieces can reflect the electromagnetic properties of the plate under measurement . Howevero 16.7o  . This pho 16.7o  ,55. The inverse methods, closely related to the forward model, retrieve the plate properties and parameter distributions, for example, conductivity profile and defect shape from the measurements. Even though the forward model varies from different governing equations and object geometry, similar inverse methods have been widely employed for a variety of objects under measurement, including pipes and spheres. The notable implementations of inverse methods in EC testing are summarized in The inversion of physical properties is realized through optimization by minimizing the discrepancy between the measured and calculated electromagnetic measurements. A least square index is usually adopted as the objective function, which represents the Euclidean distance between the measured and calculated values. For instance, the objective function employing the coil inductance reads:However, the inverse problem suffers from ill-posed and ill-conditioning problems which make the optimization extremely sensitive to measurement noise. To stabilize the optimization, the regularization methods originated from Tikhonov impose the prior knowledge as a regularization term to Function (18), which functions as a spectral filter. The objective function becomes:The measurements in the objective function can be the impedance of the Rx coil  and magnThe combination of optimized variables influence the optimization performance. It is expected that the nonlinear objective function reaches its global minimum in the variable space, which only exists when the objective function is convex. This corresponds to the situation that ductance ,65,66 anductance , the conductance . The appductance ,69. The ductance . If therIn the optimization process, the variables can be updated by various strategies according to the gradient of the nonlinear objective function. The representative methods and their characteristics are shown in For a small number of optimized variables, the heuristic methods which employ various solution searching strategies, including classic genetic algorithm (GA) and particle swarm optimization (PSO), are usually implemented in conjunction with other methods. These methods decrease the influence of starting point selection meanwhile improve the algorithm’s convergence and maintaining its correctness as long as the heuristic is admissible ,72.In previous studies, the electromagnetic properties and thickness of metallic plates have been estimated by the modified Newton-Raphson method . The afoEC testing techniques are widely applied in various industrial applications . In pracAlthough much work has been carried out for EM system development, there is still a further expectation in improving hardware and software system design. With the development of technology and higher demand in the industry, the hardware and the software have kept advancing. The development of analogue EM systems has a long history, which relies on constructing physical circuits by using electronic components as in ,90. CompThe EC testing instruments have been developed for various measurement applications. The recent studies and system characteristics are summarized in The EC testing instrument developed in the EM sensing group led by W. Yin at the University of Manchester is shown in A block diagram of the system architecture is shown in A built-in user interface running on the host PC was also developed. The user interface provides a convenient way to display and log data. It also allows the user to send configurations such as operating frequency, filter width, and coil pair assignments during the experiments. After measurements, the data is saved in a database format (.db), which can be directly fed into further signal processing algorithms.The instrument integrates all elements in a case with a size of 288 mm × 208 mm × 135 mm, which makes it portable for in-situ tests. There are also USB and HDMI interfaces that can connect to external devices. Notably, the instrument can operate with or without the power connection as it includes an internal battery with automatic power management and can last for more than 8 h during typical use once fully charged.Impedance is an important parameter characterizing the frequency response of EC probes. In the analytical solution of Dodd and Deeds, the inductance of Rx coil is calculated as the result in the presence of test pieces ,14. GeneAs compared with using off-the-self commercial impedance analyzers, the development of customized instruments can be complicated and time-consuming. Therefore, commercial impedance analyzers play essential roles in laboratory-based experiments of EC testing as they may have wider operating ranges and better performances in terms of accuracy and precision than customized instruments.Typically, commercial impedance analyzers implement 4-port measurement, where both voltage and current are measured. The terminal arrangements are shown in Generally, electronic components such as op-amps introduce phase shifts at different frequencies in acquisition channels. To reduce these phase shifts, calibration is normally required for the measuring system. Typically, it can be achieved by taking a set of measurements using a small permeable or conductive object in air space. As the expected signal in these cases has only a real (or imaginary) component, the phase shift can be manually adjusted for each frequency to minimize the imaginary  component. Then, the obtained phase for each channel can be stored and subtracted automatically from all posterior measurements using other objects . A conduAn alternative calibration method using a current sensing transformer was proposed in , where bThe EC probes can be categorized according to the operation mode and configuration of applications ,105, as For the operation modes, the absolute mode usually measures the absolute impedance of Rx coil, which reflects the thickness, conductivity, permeability, and defect information; the differential mode measures the defect on the test pieces using the differential signal of coils that is insensitive to the slowly varying properties and environmental influences; the reflection mode conventionally referred to as driver/pickup mode adopts the Tx and Rx coils which are optimized separately for various purposes.In the reflection mode, the Tx and Rx coils are considered separately. There exist various structures of Tx coils, for example, pancake, tangential, planar, and U-shaped/U-cored coils. The basic structures of several representative Tx coils and sensitive regions on measured plates are illustrated in To measure thick plates and pipes, the EC penetration depth is limited by:The defined penetration depth corresponds to the planar wave excitation and is usually higher than that of the practical Tx coils in finite dimensions. The normalized standard penetration depth can represent the influence of variables in Equation (24) , i.e., eThe U-shaped coil usually contains a ferrite core that constitute the magnetic circuit and concentrates the magnetic field beneath the two ends, which increases the signal amplitude caused by the defects ,114,115.The Rx elements include coils and magnetic field sensors which measure the coil impedance and magnetic field, respectively. In comparison, the coils are characterized by their linear response, no saturation, small hysteresis, and high flexibility, while the magnetic field sensors, for example, Hall sensor, anisotropic magneto resistor (AMR), giant magneto resistor (GMR), tunneling magneto resistance (TMR), and superconducting quantum interference device (SQUID), are in small dimensions and capable of low-frequency measurement in the sensitive direction. Nevertheless, the magnetic field sensors are limited in the dynamic range, to some extent, and provide the nonlinear response signal. The detailed characteristics and comparison of the magnetic field sensors are summarized in the literature ,124,125.Probe shielding reduces the interaction of the exciting magnetic field with the nonrelevant region in proximity of the probe, which is usually applied to reduce edge effects. In addition, probe loading employs the ferrite cores which are ferromagnetic concentrating the magnetic field for the sensing area. These strategies are significant, especially for remote field EC testing  and for The EC probes can integrate with the sensors of other sensing modalities, for example, capacitance electrodes. An integrated sensor can obtain complementary information and is sensitive to more electromagnetic properties of the test pieces. A dual-modality inductive and capacitive sensor facilitates plate thickness measurements using the single frequency inductance , as the The EC testing of planar structure suffers from the lift-off effect, which attenuates the magnitude and changes the phase of measured signals, causing errors in property estimation. To address this issue, researchers have investigated a range of methods including compensation strategies, algorithms, and lift-off invariance phenomenon.Analytical models are frequently adopted to estimate plate properties, due to explicit expressions and fast calculations. Although asymmetric problems exist in applications, a variety of cases of planar structure measurement can be approximated using the axial-symmetric model employing the Dodd and Deeds model with adequate accuracy.In the Dodd and Deeds model, the plate term of the coil inductance is a function of excitation frequency, spatial frequency, and plate properties. In the approximate model expressed by Equation (5), the characteristic spatial frequency For the nonmagnetic plates, according to the model in Equation (5), the phase of inductance is determined by the plate term ified to :(25)ϕRe[ϕThe relative permeability can be estimated if lated by :(31)α0r=For the plates in a large radius, according to Equation (8), the inductance magnitude can be calculated by:In Equation (32), the linear relationship between the lift-off Rx coils . CombiniRx coils ,114,126,Rx coils ,132. TheIn addition, it is observed that a series of quadratic-like curves related to the plate thickness can approximately describe the relationships between the phase and logarithmic amplitude performing the lift-off scan with the inductor–capacitor (LC) resonant probe. The plate thickness can be estimated employing the curve fitting method .A variety of algorithms have been proposed to address the influence due to probe lift-off on the electromagnetic property estimation of planar structures. W. Yin et al. proposed the characteristic frequency which relates to the thickness and conductivity of the planar plate . FurtherFor heterogenous plates, the material property profiles have been reconstructed from frequency sweeping measurements ,58,138. Measurements based on the electromagnetic induction method are sensitive to probe lift-off. In an investigation attenuating the lift-off effect, the lift-off invariance phenomenon was observed and applied to measure the conductive plates. As shown in With the aid of LII and LIF, optimization methods have been applied to calculate plate properties.Under high excitation frequency, it has been proposed that there exists a linear relationship between the ratio of inductance change and the probe lift-off, namely the dual-frequency linearity of lift-off (DFL) feature . This caIt has been noticed that there was a lift-off invariance inductance (LII) for different samples under various excitation frequencies, termed the lift-off invariance frequency (LIF) . The estThe defect evaluation is one of the major applications of the EC system in metallic plate measurement. The measurement features of signals in the time and frequency domain are extracted to infer the defect information. The recent research highlights are shown in The probes suitable for the surface defect measurement include the Tx and Rx coils as well as magnetic field sensors. The measured time series and frequency spectrum when scanning the probe above the test pieces, as shown in The defect information can be obtained from electromagnetic measurements and corresponding statistical features. The electromagnetic measurements are measured from coils and magnetic field sensors. The statistical features are obtained from the electromagnetic measurements through the linear and nonlinear feature extraction methods of machine learning and pattern recognition, which is intended to be informative and nonredundant, facilitating the subsequent estimation of plate properties and defect information.The measurements of coil at the characteristic frequencies, for example, peak and zero-crossing frequencies, contain the information of plate properties and probe lift-off, while the pattern of measured magnetic field parameters relates to the geometry of defects. It has been observed that the crossing frequency of a specific real part of the coil inductance corresponds to the permeability, which relates to the hardness of magnetic materials . The lift pieces . The dist pieces , i.e., pt pieces . Furthert pieces ,154, sinThe statistical features of measurements in lower dimensional space can be extracted employing the linear methods including principal component analysis (PCA), partial least squares (PLS) , clusterSpecifically, the discrete defect depth and lift-off can be estimated by classification methods, for example, support vector machine (SVM) and other ML models, implementing the classification loss functions, while the continuous dimensions and shape of defects can be obtained by the regression models .Although the emerging deep learning (DL) models have demonstrated advantages regarding defect reconstruction and lift-off tolerance , the robIn this review, we summarize the recent developments of EC testing for planar structures. The key issues in computational models, measurement systems, and parameter estimation are summarized. In the theoretical analysis, the forward and inverse problems are described, elaborating the basic principles of the analytical model and FEM for the forward problem as well as simplification and optimization algorithms for the inverse problem. The system development consists of a customized and commercial grade impedance measurement system including the calibration method, and probe design which is task specific. In applications of plate property measurement, the lift-off compensation methods are addressed. Furthermore, in defect scanning, the joint application of probes, measurement feature extraction, and defect parameter inference methods are investigated.Computational models are being developed for complex geometry and real-world applications, while, at the same time, simplified methods increase the speed of computation for real-time measurements. The sensitivity and spatial resolution of measurements can be improved by system improvements as well as customized and application-specific probe optimization. Lift-off tolerance/elimination methods are still topics worth investigation. In summary, the combination of cutting-edge models and high-performance measurement systems is still required and needs to be explored through continuous efforts for many existing and potential industrial applications.


Hand-foot syndrome (HFS) is a common adverse effect of anticancer therapy. It isknown to cause dermatological symptoms including acral erythema and dysesthesiaof the palms and soles of the feet, swelling, pain, itching, and scaling. Somedrugs, like capecitabine, are known to trigger this condition. However,pigmentation of the oral mucosa is a rare adverse effect. This study aims toreport a case of oral mucosa hyperpigmentation caused by capecitabine therapybefore the clinical diagnosis of HFS. A 58-year-old female, diagnosed withinvasive breast duct carcinoma, had the central nervous system, liver, skin, andlung metastasis, using capecitabine every day for 14 cycles. Oral examinationrevealed multifocal black macules on the hard palate, bilateral buccal mucosa,gingival mucosa, and dorsum of the tongue. The clinical hypothesis was oralmucosa hyperpigmentation by capecitabine use and only periodic follow-up wasnecessary. Hyperpigmentation of oral mucosa by capecitabine is a rareconsequence of neoplastic therapy and your association with HFS is unclear, andpoorly reported. The report of these events is important to alert oncologyhealth teams about the individual tolerance to capecitabine therapy. In addition, previous studies have questionedwhether hyperpigmentation of the oral mucosa is yet another manifestation of HFS orwhether it behaves differently A 58-year-old black female, with a history of alcoholism and smoking forapproximately 30 years, and abstinence for 18 years, attended the dental office at areferral hospital, referred by the radiotherapist in May 2018, complaining ofdryness of the oral mucosa. Through intraoral physical examination, there weremultiple blackened - dark macules in the hard palate, bilateral buccal mucosa, upperand lower gingival mucosa and dorsum of the tongue , however2N3Mx) in2015. She was initially submitted to unilateral mastectomy and axillarylymphadenectomy, followed by adjuvant chemotherapy using the ACT protocol until November 2015, followed byradiotherapy (RT), 5040cGy (28 fractions of 180cGy in right plastron and 4500 cGy in25 fractions of 180cGy in the right clavicular fossa), from March 2016 to April2016. The ACT protocol was suspended due to disease progression and therapy withoral chemotherapeutic drugs, tamoxifen and anastrozole was instituted, associatedwith RT in the plastron region. Two years after the initial diagnosis, she presentedwith disease recurrence in the left orbit and in the plastron, being subjected to anorbitectomy. In 2018, the patient was diagnosed with metastasis in the centralnervous system and liver, when she underwent a new RT in the head and neck region.In the same year, two new metastasis were found in skin and lungs, when oralcapecitabine (XELODA®) was prescribed. The recommended dose was 500mg / twice a dayfor 14 days, followed by a rest week. After 14 cycles, she showed low tolerance totherapy with asthenia and diarrhea. For that reason, the drug was suspended toadjust the dose, and then, the patient was diagnosed with grade 1 HFS by theoncologist. At this time, she attended the Dental office, when hyperpigmentation ofthe oral mucosa due to chemotherapeutic agent diagnosis was closed, a condition thatwas present before the diagnosis of HFS by the medical team. From the diagnosis, theconduct established by the dentistry team was the periodic clinical monitoring ofthe oral spots. The patient was followed for a period of 22 months and died due tomalignancy in July 2020.The patient´s medical history consists of a diagnosis of invasive ductaladenocarcinoma on the right breast  classifies the disease in three stages accordingto the level of severity as described in ,,The pathophysiology of HFS remains a matter of controversy. Several studies havetheorized mechanisms that seek to explain the disease. Sanghi et al. ,,,,,There are currently few cases of capecitabine-induced hyperpigmentation published inthe literature ,,,,,The possibility that oral mucosa pigmentation be a distinct entity or that might beother manifestation of HFS has been the subject of discussion in the literature,because of the few reports describing oral mucosa pigmentation associated with HFS,,The diagnosis of drug-related hyperpigmentation is based on the relationship betweenthe onset of signs and symptoms, the time of drug administration, and based on theexclusion of lesions that have a similar clinical appearance, but with a differentetiology ,The treatment of HES is basically symptomatic through the prescription of topicalagents. The use of urea or lanolin-based protective ointments and the use of mildemollient creams is advisable. Other supportive measures such as the administrationof topical and systemic antibiotics and the use of topical corticosteroids todecrease pain and inflammation may also be used. Despite this, the most appropriatemanagement for the syndrome is the discontinuation of the drug. Complete healing ofHFS is possible as long as localized ulceration does not occur Due to the clinical appearance of this lesion, the differential diagnosis with otherpigmented lesions in the oral cavity is essential, and requires the involvement ofthe dentist in the multidisciplinary oncology team. Knowing the patient's medicalhistory and understanding the oral repercussions of the various drugs used asantineoplastic therapy is the dentist's role, especially in cases of oralpigmentation where no or few therapeutic approaches are availableHyperpigmentation of the oral mucosa is a rare consequence of capecitabineantineoplastic therapy. Whether it is a part of an early manifestation of HFS or ifit is an independent entity remains unclear. We report a case of hyperpigmentationof oral mucosa before the clinical diagnosis of HFS, which could be an early sign ofthis syndrome. New studies should be published in the literature in order toelucidate this process.


Varieties of microorganisms reside in the oral cavity contributing to the occurrence and development of microbes associated with oral diseases; however, the distribution and in situ abundance in the biofilm are still unclear. In order to promote the understanding of the ecosystem of oral microbiota and the diagnosis of oral diseases, it is necessary to monitor and compare the oral microorganisms from different niches of the oral cavity in situ. The fluorescence in situ hybridization (FISH) has proven to be a powerful tool for representing the status of oral microorganisms in the oral cavity. FISH is one of the most routinely used cytochemical techniques for genetic detection, identification, and localization by a fluorescently labeled nucleic acid probe, which can hybridize with targeted nucleic acid sequences. It has the advantages of rapidity, safety, high sensitivity, and specificity. FISH allows the identification and quantification of different oral microorganisms simultaneously. It can also visualize microorganisms by combining with other molecular biology technologies to represent the distribution of each microbial community in the oral biofilm. In this review, we summarized and discussed the development of FISH technology and the application of FISH in oral disease diagnosis and oral ecosystem research, highlighted its advantages in oral microbiology, listed the existing problems, and provided suggestions for future development.. The human oral microbiota consists of more than 700 species, and the microbial communities are diverse at different niches in the oral cavity . Human oHuman oral microbiota has distinct characteristics ,6. The hVarious oral microbiological detection methods have been developed in labs and clinics . FluoresFISH is one of the most routinely used cytochemical techniques for genetic detection, identification, and localization by a fluorescently labeled nucleic acid probe. It is more specifically used in hybridizing processes with nucleic acid sequences of interest . The vitIn the 1960s before the advent of FISH, cytochemical methods of detecting and localizing specific intracellular molecules were mainly immunocytochemistry based on in situ hybridization with radiolabeled probes and antigen-antibody interaction with fluorescein-labelled immunoglobulins ,21. HoweAs a combination of in situ hybridization and indirect immunofluorescence, a method described by Rudkin et al. in 1977 replaced autoradiography with fluorescence microscopy to detect in situ signals from antibodies against DNA-RNA hybrids . Before The optimization of probe design and synthesis is crucial to the development of FISH. The establishment of genetic databases made it possible to design probe sequences targeting complementary gene sequences of interest . Probe pWith accurate probe design methods, FISH properties became even more modern, with changes that include increasing signal intensity and stability, range of targets, and sensitivity. Commonly used variants are: catalyzed reporter deposition FISH (CARD-FISH), gene-FISH, recognition of individual genes FISH (RING-FISH), nucleic acid mimics FISH (NAM-FISH), combinatorial labeling, and spectral imaging FISH (CLASI-FISH), double labeling of oligonucleotide probes for FISH (DOPE-FISH), as described below. The polynucleotide probe also has an advantage in signal intensity with multiple labels and secondary structures mediating probes-connected networks . RING-FIThe nucleic acid mimics probe has become the most promisingly high-efficient probe with higher affinity, specificity, and better stability based on resistance to enzymatic degradation , used toThe variants mentioned above show corresponding advantages in detecting microorganisms, allowing direct identification without cultivation and further detection of microbial community structure and individual function ,38. As mThe variants of FISH are improving, as are their applications based on the detection of nucleic acids. Apart from microbiological applications, FISH has been a gold standard technique in absolute gene copy number quantification in cancer, allowing implementation of precise treatment strategies . Based oThe general protocols of FISH include (a) specimen treatment, (b) probe denaturation, (c) hybridization, (d) elution, (e) hybridization signal amplification (applicable to biotin-labeled probes), (f) re-staining, (g) encapsulation, and (h) fluorescence microscope observation of FISH results ,57,58. P6 cells per well; For environmental bacteria: sludge samples or sludge particles and other sample forms that can be used for smear or sectioning [FISH can evaluate various types of test samples, such as tumor tissue, pathological sections, local animal samples, human tissue samples, etc. For tissue samples: 4% paraformaldehyde fixation (paraffin section), or put into liquid nitrogen, −80 °C storage, sample not less than 100 mg; For cell samples: cell slides prepared with 6-well plates, with no less than 2 × 10ctioning ,64,65. Though FISH is widely used in microbiological detection and biofilm analysis, some techniques belonging to nucleic acid, immunity, and single-cell techniques also have been applied in different emphases of microbial research due to their different advantages. Ten common techniques (including FISH) in microbial research were selected and compared based on several performance factors, including detectable resolution, applications in oral microbiology, culture reliance, quantification appliance, capability for microbiota analysis, and unsearched species . In summFurthermore, most oral microorganisms are unculturable, and the community structure and components are complex and changeable, and these properties make FISH very suitable for oral microbial research. Among many variants of FISH, classical FISH and CLASI-FISH are the most popular in terms of their application in oral microbiology. However, it is noteworthy that a new variant called HiPR-FISH is considered to have a promising future. The procedure and principles of these three currently used FISH techniques in oral microbiology will be elaborated throughout this study.The most common procedure followed by researchers who apply FISH involves the following four steps: (1) fixation for dehydration to inhibit the action of enzymes without cell and nucleic acid structures destruction ; (2) hybridization between target nucleic acid sequences and specific complementary probes labeled with fluorescent dyes or reporter molecules to be detected by fluorescent antibodies; (3) washing to remove the unbound or loosely bound  probes; (4) detection and visualization of the probe-bounded cells by typically fluorescence microscopy and some advanced microscopy systems, then access to information for species detection or microbiota analysis ,36,87. HRegardless of some minor adaptions to classical FISH protocols for CLASI-FISH application, the main improvement of CLASI-FISH are probe design and image acquisition/analysis. CLASI-FISH probes are particularly effective in synthesizing two probes for one targeted sequence and two different sequence-particular fluorophores; thus, the cells of different species are labeled by specific combinations of two highest-intensity fluorophores . For detRegarding the expansion of the number of distinguishable species in a single image as CLASI-FISH, it has been previously verified that High Phylogenetic Resolution FISH (HiPR-FISH) achieves significantly higher taxonomic resolution and multiplexity due to a two-step hybrid approach and a routine for automated image segmentation . Two kinTen distinct fluorophores encode up to 1023 fluorophore combinations, and one species corresponds to a 10-bit binary barcode derived from designed one or more encoding probes binding with relevant readout probes and presenting different spectral components. After image acquisition, automated image segmentation classifies the spectra of images and assigned cells the corresponding barcodes. The reference spectrum for each barcode is established by the Förster resonance energy transfer (FRET) model, and the k-means clustering-denoised and straightened images of cultured single-cell and biofilm samples are segmented by the watershed algorithm that the seed is defined by developed Local Neighborhood Enhancement (LNE). For spatial analysis, the adjacency segmentation is also generated by the watershed algorithm and then calculates the intuitive spatial association matrix. The super-resolution images and 3D datasets are generated the same way, and the 4D data cube needs rendering in ipyvolume.FISH has been widely used in various fields of oral microbial research. From the basic research at the practical level to the potential clinical application and outcome transformation, the important role of this methodology has been witnessed by scientists in different scenarios inside the research community. In this section, we summarize the three major applications of FISH and explain its relevant details .This function of FISH has been used to identify the species of pathogenic microorganisms in the simplest way, but it is still in the basic research stage in terms of predicting the risk of disease and prognosis, which needs to be further tested and verified in clinical practice. In the context of modern medicine, the requirements of patients and clinicians for diagnosis have upgraded from merely knowing the type of diseases to detecting specific causes and predicting disease progression. For patients with diseases related to oral microorganisms , most of them exerted evident symptoms at the time of its’ initial diagnosis, such as pain, inflammation, and changes in biological characteristics. And increasing studies have shown that cancer, atherosclerosis, diabetes and other diseases can be reflected in oral flora. Oral microorganisms play an important role in the occurrence and development of these diseases, and the changes in community structure can somehow indicate how it has been progressing . TherefoClinically, FISH can be used to detect and analyze various disease-causing factors, such as HIV and Epstein-Barr virus, providing a faster way to diagnose such conditions. However, due to the spreading of the new COVID-19 pandemic, nucleic acid testing has become a part of people’s lives, so we urgently need a more rapid and more convenient way to detect the virus. Hepp et al. developed a rapid FISH protocol capable of quantitatively detecting influenza virus, avian infectious bronchitis virus, and SARS-CoV-2 in approximately 20 minutes by combining nasal and throat swabs with the added virus in virus cultures. This rapid and simple method can be used both as a commonly used detection technique in the laboratory and to aid the future diagnosis of enveloped viruses with accessible genomes . Modern research shows considerable differences in the type, proportion, and structure of oral microbiota between healthy and pathological states, which can be used as breakthrough points to diagnose the actual causes of the disease. FISH was initially used to detect relatively single and known pathogenic microorganisms, but with the development of new technology, a variety of microorganisms can now be detected simultaneously. Clinicians and researchers use FISH to detect microbes’ information, such as species, and community structure, after extracting samples from patients. This kind of data is included in the databases and also used as feedback to clinicians to develop more accurate treatment plans. Taking oral lichen planus (OLP) as an example, there are significant differences in microflora between OLP patients and healthy controls. By applying FISH, Zheng et al.  and WangStreptococcus spp., and F. nucleatum using specific fluorescent probes. In their experiment, they found a significantly higher proportion of F. nucleatum and Streptococcus spp. in the biofilm of the halitosis group. It was concluded that the relative ratio of total microorganisms to these two bacteria could be considered a relevant factor in causing bad breath; hence it can be included in the treatment goals [In addition to being used in the diagnosis of diseases, the results of FISH can be used in the formulation of treatment options. Take the most common and inconvenient form of bad breath as an example. Bernardi et al. visualized and quantified the dorsal biofilm through the combination of FISH and CLSM and stained eubacteria, nt goals .Apart from oral diseases, FISH is also used for microbial detection of lesions of other diseases. For example, it has been found that there are many common oral bacteria in oral squamous cell carcinoma . AlthougCurrently, research focus on oral microecology occupies an important position in stomatology , and manAccording to the Expanded Human Oral Microbiome Database (eHOMD), only 57% of oral microorganisms can be cultivated and formally named, 13% of oral bacteria can be cultivated but not named, and 30% of the oral microorganisms fail to be separated from the biofilm, hence hindering the study of oral micro-ecological research . FISH, wAnnett et al. detected the presence of Treponema pallidum in the tissue samples of syphilis patients by a specific 16S rRNA FISH probe against T. pallidum. Their effective methodology avoided possible false results of serological identification in HIV-positive patients and provided a faster and more effective technique for disease monitoring . Using sDue to its ability to display location information apart from identifying species, researchers have increasingly applied FISH to monitor microbial communities in recent years. The technique can be used to study colonies that are difficult to cultivate and clearly present the overall microbial environment without requiring additional bias-prone steps such as extraction and amplification. FISH can accurately and quickly reflect the in-situ distribution of microbial communities in various systems, avoiding the complex process of other community analysis methods . ThroughBring et al. spotted the distribution of TM7 bacteria, which is difficult to culture in the oral microecology through FISH and further reveals the mechanism of their metastatic infection . Shi et Using the combination of FISH and CLSM, AI-Ahmad et al. measured microflora changes in oral plaque within eight weeks with a particular focus on individuals with poor oral hygiene . CombiniBecause of its two effective features for species identification and community detection, FISH is often used to study dental plaque biofilm. Mature dental plaque consists of multiple species of biofilms and contains more than 500 different bacteria, causing common oral diseases such as periodontitis, dental caries, and gingivitis . CombineThe purpose of studying oral microorganisms is to maintain a healthy oral microecology, so scientists make efforts to establish a fairly complete effectiveness evaluation system. At present, the research topics related to using FISH as one of the detection methods mainly include molecules, oral materials, and the establishment of models and methods.In the long history of people dealing with oral pathogens, researchers have found that some molecules can specifically inhibit some of these pathogens and promote the reproduction of probiotics. The first kind of molecules are part of our daily lives, and it is commonly known that some have medicinal properties, such as salicylic acid, podophyllotoxin, and vinblastine. Hannig et al.  suggesteS. mutans, manifesting that LCG-N25 could be a hopeful adjuvant for the treatment of caries.The second kind of molecule is related to the artificial synthesis and improvement of natural compounds. Lyu et al.  studied After exploring the role of individual molecules, our team identified that it was equally important to understand the effects of the interaction of different molecules on our results, so a new series of experiments was carried out. Cheng et al.  investigS. mutans [S. mutans [In addition to FISH, other technologies are also widely utilized, such as using PCR to explore the effects of terminalia chebula extracted from ethylalcohol on . mutans , using D. mutans , and so Streptococcus spp. content in the biofilm, revealing that the surface roughness of the zirconia surface of the oral implant with low roughness was similar to that of titanium surface in terms of initial bacterial adhesion or oral Streptococcus spp. content in the biofilm. Similarly, PCR was used to unveal how TiO2 in the form of nanotube inserted into GIC affect S. mutans [The oral cavity is an essential human structure closely related to food nutrition absorption, gas exchange, and speech expression. Consequently, dental materials are required to have biosafety, biocompatibility, and biofunction. However, the biofilm formed by microbes attached to the material surface can lead to drug-resistant infection and the contamination of medical devices . Accordi. mutans , but it The model method is a bridge between scientific theory, an effective means of successful scientific research, and a carrier for the sublimation of creative thinking. The research community has been exploring this method and expanding the knowledge about it each day, shedding light on the crucial role it can play in modern medicine. Many important experimental results have been achieved via teaching models in the classroom, experimental animal models and cell models, bioreactors, computer-aided 3D modeling, and mathematical modeling. At present, the model research on oral diseases caused by oral microbes can be divided into in vivo and in vitro models. For in vivo models, more clinically significant indicators such as plaque index and alveolar bone resorption are often used to judge whether the model is successful or not. As for the in vitro models, especially when the results are verified by FISH, the bioreactor is widely used to simulate the physiological or pathological environment. Different models, including an oral biofilm model of dental pulp disease established on hydroxyapatite and dentin disc , a repeaIf the model method is an up-to-date summary of the major findings of systematic research, then the establishment of the new methodology represents the improvement or subversive creation of the existing methods. However, the improvement or innovation of oral microbiological research methods requires more convincing proof methods, e.g., conventional slide detection, immunological methods, and molecular biology techniques , which mIn recent years, FISH has significantly impacted clinical and microbiological detection methods due to its high specificity and efficiency, making it one of the most promising methods for clinical microbiological detection. In addition, FISH is commonly used in daily life experiments and may become a standardized experimental method in future research. Multiple optimizing standpoints of FISH have spawned numerous variants. Based on four fundamental procedures, innovations in probe design have been successfully put into practice, greatly expanding the applications of this methodology. Regarding potential applications, the specific fields are featured with diagnosis assistance, oral microecology research, and effectiveness evaluation, manifesting the great applicability of FISH in oral microbiology. Although the multimolecular affinity and good biocompatibility of FISH make it capable of providing a new and effective method for detecting oral microorganisms, the method still has its shortcomings, especially when it comes to potential interference with false-negative results due to signal loss. Albeit multiple variants of FISH have been improved from several perspectives, the ensuing more complex procedures and higher demands on experimental manipulation and equipment limit their practical applications. For basic FISH, the probes present the problem of the inability to achieve 100% hybridization rate, especially with cDNA probes which are complementary to mRNA. The detection of mRNA is mainly used for transcriptomic analysis but the level of mRNA does not fully reflect the level of expressed proteins. In addition, a lack of standardized processes for the analysis of FISH results exists, which leads to the need for experienced analysts to ensure relatively correct judgment, which undoubtedly brings extra training costs. At present, some disease sites only use detection technology to confirm the existence of oral microorganisms, and most of the research on FISH detection of oral microorganisms focuses on the impact on oral diseases, lacking research on other diseases that have been proved to be related to oral microorganisms. Therefore, FISH and its variants can be used in the future to analyze the community structure, dynamic evolution process, interaction with other cells and the relationship with disease occurrence and development of oral microorganisms in these lesions, so as to help doctors and researchers better develop and use drugs and other interventions.With the advent of the era of precision medicine, microbiome and its precise regulation are exceedingly important in the research of microbial-related diseases. Thereby, the common application with other technologies and the innovation of FISH itself has become a general trend within the scholarly community, and the emergence of CLASI-FISH and HiPR-FISH are relevant examples. The emergence of these new technologies enables us to understand the interaction among genes, metabolites, and signal pathways in the oral microbial community from an ecological point of view rather than the previous relatively single interpretation .Altogether, the future direction of FISH can be divided into two categories: (1) The improvement of FISH itself. For example, the further improvement of the accuracy of probes, the way to expand the variety and total number of probes while reducing the interference effect, and cost reduction. (2) The cooperative application of FISH and other technologies. For instance, to develop a full-process automated FISH experimental device by combining the image recognition technology of machine learning and deep learning and using FISH to analyze the three-dimensional models of oral microorganisms with real-time and dynamic monitoring. Hypothetically, if the ideas get actualized and widely accepted in the future, problems that need to be discussed from the perspective of “microorganism-host-system” can be studied more conveniently, quickly, and accurately. For instance, the establishment of a dynamic competitive ecological model of dominant bacteria in oral cavity, study on the evolution of pathogenic bacteria in hotbeds such as dental plaque, and the potential ways of oral pathogens to invade other parts of the body. Based on the dynamic three-dimensional database of oral microecology obtained by FISH, we can further explore the fundamental relationship between oral microorganisms and tumors, atherosclerosis, Alzheimer’s disease and other diseases, so as to develop monitoring equipment for oral microbial detection based on FISH probes to cooperate with other auxiliary tests to predict the risk of disease, determine the degree of disease development and judge the prognosis. This flourishing research field will scale to new heights based on these achievements.