Lemuridae in response to various types of potential stressors during their stay in a temporary housing facility. The glucocorticoid levels were specifically monitored for ring-tailed lemurs (Lemur catta), white-headed lemurs  and collared brown lemurs (Eulemur collaris).Lemurs kept in captivity are constantly influenced by a number of factors that affect their welfare. All lemur species are classified as endangered species according to the International Union for Conservation of Nature (IUCN) Red List and their population in the wild is decreasing. It is therefore useful to improve the methods for assessing the level of stress in individual lemurs in captivity. In this study, we compared the changes in glucocorticoid concentrations in three species of p < 0.05) the FGM levels within one to two days after the event. Although housed in the same room, differences in response to potentially stressful situations were found in white-headed lemurs compared to ring-tailed lemurs. Increased mean levels of the FGMs were found more frequently in white-headed lemurs than in ring-tailed lemurs. The results suggest that this species may be more sensitive to changes in its surroundings. In general, the levels of the FGMs showed a similar pattern during 30 days of monitoring suggesting that all groups of lemurs responded in a similar manner to the same events. However, we recorded the differences in the absolute values of glucocorticoid concentrations between the monitored species likely due to the differences in sex ratios in the groups and presence of lactating females in the ring-tailed lemurs.We compared the glucocorticoid concentrations in response to various types of potential stressors present during standard operation of a temporary housing facility between three species, namely, ring-tailed lemurs, collared brown lemurs and white-headed lemurs. The levels of faecal glucocorticoid metabolites (FGMs) were measured non-invasively on a daily basis during a 30-day period. A total of 510 faecal samples were collected. Concentrations of immunoreactive glucocorticoid hormone metabolites were measured in the obtained extracts by using an enzyme immunoassay. The polyclonal antibodies used in this assay were directed against the metabolite 11-oxo-etiocholanolone I. We found all three monitored lemur species to respond to specific potentially stressful situations by increasing ( Animals kept in zoos are constantly influenced by a number of factors that affect their welfare. In order to ensure the welfare of captive animals, it is essential to avoid excessive and, in particular, prolonged stress . Even geSeveral methods are commonly used to evaluate stress response. They are largely based on the determination of glucocorticoid hormone levels of cortisol and corticosterone or their metabolites ,4. A comPongo pygmaeus) and Sumatran orangutans (Pongo abelii). Higher levels of glucocorticoids in the Bornean orangutans were predicted, due to the much more solitary lifestyle and higher frequency of male aggression directed towards females in this species in the wild, which was subsequently confirmed [Reactions to stressors often vary among different animal species ,9,10. Deonfirmed . Furtheronfirmed  found thVarecia variegata) are reported to have a fast passage of the digestive tract (2 to 4 h). Given the very similar structure of the digestive tract in ring-tailed lemurs (Lemur catta) [Eulemur collaris) living in fragments of forests with a group living in better preserved areas. For the determination of glucocorticoid hormones in the faeces, a successfully validated analysis for this species targeted against the metabolite 11-oxo-etiocholanolone was chosen. Similarly, glucocorticoid levels in faeces during the reproduction season were analysed in red-fronted lemurs (Eulemur fulvus rufus) [To achieve reliable results via non-invasive monitoring in individual species, knowledge of its metabolism and the mechanism of glucocorticoid hormone excretion is essential . Accordir catta) , the timr catta)  compareds rufus)  using ths rufus) ,19.Lemuridae are generally classified as endangered species according to the International Union for Conservation of Nature (IUCN) Red List and their populations in the wild are decreasing although they are frequently kept in captivity. In order to ensure their stable population, health and welfare in captivity, it is important to reduce the negative effects of stress. A commonly measured endocrine response to stress is the secretion of glucocorticoids [rticoids . Consequrticoids ,10.Lemuridae, namely ring-tailed lemurs (Lemur catta), white-headed lemurs  and collared brown lemurs (Eulemur collaris). Given their small body size and fast metabolism, changes in FGMs were expected to be measurable within one to two days after exposure to a stressor [Pongo spp.) [Callithrix jacchus), long-tailed macaque (Macaca fascicularis), Barbary macaque (Macaca sylvanus), chimpanzee (Pan troglodytes) and gorilla (Gorilla gorilla) [Gorilla gorilla gorilla) [Hylobates pileatus) [Lemur catta) [Varecia variegata) [The aim of this study was to compare the changes in glucocorticoid concentrations in response to various types of potential stressors occurring during a 30-day period in a temporary housing facility in the three selected species of stressor ,10 as cogo spp.) , common gorilla) , Westerngorilla) , pileateileatus) , wild grileatus) , ring-tar catta)  and blacriegata) . Lemuridae in an approved holding facility. Since only non-experimental clinical veterinary practices were performed and no handling of animals related to research was carried out, a formal ethics approval from the Animal Welfare Body of the University of Veterinary and Pharmaceutical Sciences Brno with regard to the EU Directive 2010/63/EU was not required.This study was carried out in strict accordance with the Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes and Czech national legislation  on the protection of animals against cruelty, as amended. All samples were collected non-invasively and animals did not undergo any experimental procedures. Sampling was carried out during temporary housing of three species of For the analysis, faecal samples from 9 ring-tailed lemurs, 6 collared brown lemurs and 2 white-headed lemurs were collected for a period of 30 days. The lemurs originated from European zoos and were temporarily housed in a holding facility in Poštovice in the Czech Republic operated by a certified international transportation company, The Nature Resource Network, that was in charge of their transport to other zoos. Details on individual animals monitored in the study are given in All lemurs were fed primate dry formula (pellets), vegetables  and fruits  two times in a day, with green leaves, and with cheese and boiled egg once a week. Drinking water was available ad libitum. The lemurs were housed in indoor enclosures  without access to an outdoor area. Changes in the groups during the monitored period are described below. In ring-tailed lemurs, 9 adult individuals were monitored . There were 4 offspring in the ring-tailed lemurs but they were not included in the analysis. Four adult females (R1–R4) were housed in cage 1 in room 1. Two of them had offspring . The two lactating females (R3 and R4) together with their offspring were separated on day 16 of monitoring to another cage in the same room due to their agonistic behaviour. Female R3 with her offspring was housed in cage 2 and female R4 with her offspring was housed in cage 3. The remaining two adult females (R1 and R2) were left in cage 1. Subsequently, female R1 was isolated in a separate cage 4 in the same room due to injury on the 23rd day of observation. Another four adult ring-tailed lemurs (R5–R8) were housed in room 2. Each adult was individually housed in separate cages, except for female R6, who was co-housed with her offspring. The first day of monitoring one more adult female ring-tailed lemur (R9) arrived at the facility and was placed in a cage in room 6. Rooms 1 and 6 were interconnected. In white-headed lemurs, 2 individuals  were housed together throughout the whole monitoring period in room 1 in cage 5. In collared brown lemurs, 6 individuals were assessed: 2 males (C1 and C2) and 4 females (C3–C6). Originally, they were housed together in cage 1 in room 5. During the monitored period, however, they were divided into smaller groups due to mutual attacks and relocated to different cages within the same room (room 5). Namely, on day 16 two males (C1 and C2) were relocated to cage 2, on day 18 one female (C3) was relocated to cage 3, on day 23 one female (C4) was relocated to cage 4 and on day 29 one female (C6) was relocated to cage 5. While the ring-tailed lemurs and white-headed lemurs were housed in the same room, the collared brown lemurs were kept on the other side of the facility. A scheme of the floor plan of the facility and relocation of individuals is provided in g, 10 min). The detailed extraction process is described in other methodological publications [The monitoring was carried out for 30 days when three lemur species were kept in the facility. Lemurs were exposed to the same husbandry activities . All activities related to the operation of the facility were recorded  by the caretakers during the monitored period in a systematic way with the use of pre-prepared sheets. The faecal samples for the determination of FGMs were collected from the floor once a day between 8:00–12:00 a.m. from each animal, ensuring that they were not contaminated with urine, water or other material. A total of 510 faecal samples were taken. There was no variability in the number of samples taken from different animals. The collected samples were put in plastic sealable bags within 1 h of defecation, labelled and immediately frozen at −20 °C. The subsequent sample processing including extraction was performed at the University of Veterinary Medicine in Vienna, Austria. The samples were thawed, individually homogenised and then successively put into test tubes at weights between 0.480 and 0.520 g. Afterwards 1 mL of distilled water and 4 mL of 100% methanol were added to the weighed samples. Each sample with water and methanol was stirred on a shaker for 30 min and then centrifuged at 4 °C , based p < 0.05 was considered significant. The first day of sample collection was the day when FGM levels were expected not to be affected by stress, not even in those individuals who arrived on the first day of observation due to a time delay in excretion of FGMs [The results were statistically evaluated using the statistical program Unistat 5.6 . Data normality was tested using the Shapiro–Wilk test . Since t of FGMs . TherefoPteronura brasiliensis) were relocated to room 6 . The lemurs had auditory and olfactory contact with these two otters. Records on daily events showed a variety of activities occurring in and outside the rooms where lemurs were kept during a 30-day period of monitoring. In room 1, besides daily routine , noise from construction work was also recorded. Namely, on the 13th day of the monitored period, all-day construction work was carried out from the other side of the room where the ring-tailed lemurs were housed. The construction work was also performed on day 14. Furthermore, some females originally housed in one group in the same cage were moved to separate cages. On day 16, two females (R3 and R4) with offspring were relocated into two separate cages and on day 23, one female (R1) was moved due to an injury to a separate cage, but all stayed in the same room. On day 29, two giant otters (n = 9) over a 30-day period of monitoring are shown in (29.232) = 2.3765). On the 14th day of the monitored period, a significant increase (p < 0.05) in the mean FGM levels was found. The mean FGM levels also increased on day 17 (p < 0.01) and day 24 (p < 0.05). Subsequently, on the last day of monitoring, a significant increase in the mean FGM values (p < 0.01) was also observed. On other days, the FGM levels did not significantly differ from the first day. Fluctuations in the mean FGM levels in ring-tailed lemurs  housed together in one cage located in the same room as the ring-tailed lemurs over a 30-day period of monitoring are shown in (29.29) = 1.6832). A significant increase (p < 0.05) in the mean FGM levels was found on days 10, 15, 16, 18, 24, 26, 29 and 30 in comparison with the first day of sampling. Fluctuations in the mean FGM levels in white-headed lemurs (Macropus rufogriseus) arrived on the third day after the beginning of the observation and they were temporarily placed in the same room as the collared brown lemurs. They were housed in a different room than the other two lemur species. All of them arrived at the facility on the first day of monitoring and were initially placed together in cage 1 in room 5. During the monitored period, however, some individuals had to be moved due to mutual attacks as described in n = 6) over a 30-day period of monitoring are shown in p < 0.05) increase in the mean FGM levels (F(29.145) = 2.1247) was found on days 17, 18, 20, 24 and 30.Fluctuations in the mean FGM levels in collared brown lemurs (p < 0.05) were observed. As shown in When comparing the FGM levels between all three lemur species, significant differences  in the mean FGM levels was found. This increase could result from ongoing construction work as lemurs were exposed to increased noise and vibrations in their cages. The mean FGM levels also increased on day 17 (p < 0.01) and day 24 (p < 0.05). These increases could be related to relocations of two females with offspring (R3 and R4) into separate cages (16th and 23rd day). The separation was necessary due to agonistic behaviour observed in the females. Despite the females being familiar with each other from their original location, stress from the new environment could cause changes in their behaviour. Relocation can be perceived as a stressor in captive wild animals because they perceive all changes and new situations as a potential threat [p < 0.01) was observed. According to records, two giant otters (Pteronura brasiliensis) were relocated into the facility on the previous day which could be stressful for lemurs housed nearby. In the monitored period, relatively stable FGM levels were found on days which were not preceded by any exceptional events (daily routine was followed). Assumedly, the above-mentioned disturbances were responsible for the changes in FGM levels and they showed a consistent one-day delay before elevating FGM concentrations. White-headed lemurs were exposed to the same potentially stressful events as the ring-tailed lemurs. Construction work was in progress on day 13 and 14 near their enclosure, which could explain a significant increase (p < 0.05) in the mean FGM levels on days 15 and 16. Interestingly, there was a two-day delay before the FGMs elevated in white-headed lemurs in contrast with the one-day delay in ring-tailed lemurs following the same stressor. The significantly elevated levels of FGMs were found also in the last days of monitoring (day 26 to 29). The reason is unclear. The records did not show any deviations from the daily routine. However, the animals might have responded even to some minor changes not detected or perceived by the caretakers. According to Morgan and Tromborg [On the 13th and 14th day of the monitored period, all-day construction work was carried out near the room where the ring-tailed lemurs were housed. In the days after (14th and 15th day) a significant increase (l threat . On day p < 0.05) increase in FGM levels was recorded following the construction work throughout the day in the yard of the facility where both species were housed. As Morgan and Tromborg [In both, the ring-tailed lemurs and the white-headed lemurs, a significant  in the mean FGM levels was found on days 17, 18, 20, 24 and 30, which could be caused by aggression and subsequent relocations. Mutual fights escalated especially during feeding. Aggressive behaviour has been proven to affect FGM levels, with dominant individuals having higher glucocorticoid levels [Collared brown lemurs were housed in a different room than the other two lemur species. Thus, they could be affected by different potentially stressful events. In particular, these included necessary relocations due to the occurrence of agonistic behaviour in the group. In the monitored period, a significant increase  to the same room. Furthermore, in collared brown lemurs there was no increase in the mean FGM levels in response to the construction work carried out on the 13th and 14th day of the observation, likely because the collared brown lemurs were located on the other side of the facility, thus no noises or vibrations from the construction work were recorded here.While the significant increase of FGMs in ring-tailed and white-headed lemurs was caused the next day after the arrival of giant otters, in collared brown lemurs no significant change was found in FGM levels following the arrival of Bennett’s wallabies (Microcebus murinus) based on the determination of elevated levels of FGMs after capture. Although elevated levels have been recorded in some cases, routine capture has not had long-term consequences. The stress response in black howler monkeys (Alouatta caraya) was evaluated by Rangel-Negrín et al. [During the monitored period, handling of animals was most often associated with their relocation to other cages due to agonistic behaviour. For relocation, capture and direct handling was necessary. That can be perceived as a strong stressor ,28 with n et al. . The resn et al.  recordedn et al. . Given tp < 0.05) the FGM levels similarly within one to two days after the event. This delay from the stress event to GC excretion into faeces was confirmed not only in previous studies regarding handling of the ring-tailed lemurs and transportation of the black-and-white ruffed lemurs, but also in many other studies concerning primates [The results show that all the three monitored lemur species respond during the observed period to specific stressful situations by increasing  compared to males due to pregnancy. Dantzer et al. [Tamiasciurus hudsonicus) also have elevated levels of FGMs during pregnancy. Furthermore, Carrera et al. [Theropithecus gelada).The differences in the FGM concentrations in the individual species may be also due to different proportions of sex in the observed groups , becauser et al.  found fea et al.  found hip < 0.05) in the FGM levels within one, or at most two days after the stressor exposure. Total concentrations of the FGMs differed between the species, with the highest in ring-tailed lemurs, lower in collared brown lemurs and the lowest in white-headed lemurs. In white-headed lemurs, much lower FGM levels were found in comparison with the other two lemur species, which could be interpreted as a species difference. However, it is also possible that their low levels of FGMs corresponded to the fact that as the only one their group was not divided during the monitored period, and the males were used to each other. For social species, their mutual dependence is high [The study has shown that some situations and activities related to captive housing can result in elevated levels of FGMs in lemurs. A similar pattern of changes in the FGM levels in response to potentially stressful situations was observed in the monitored lemurs. All three lemur species in our case showed a significant increase , different number of animals and varying sex ratio in the monitored groups. However, it was not possible to experimentally design and control the structure of the monitored groups and the conditions of observation. Lemuridae suggesting that closely related animal species respond to stress load similarly. A significant increase in the FGM levels was found within one, or at most two days after the exposure to a stressor. Stress is currently one of the most serious problems in rearing endangered animal species in zoos and other facilities around the world. For their preservation and successful breeding, it is therefore of the utmost importance to avoid excessive stress and to provide the most suitable conditions for their life in captivity. In order for the housing conditions to be properly assessed, it is necessary to determine the impact of the factors to which captive lemurs are exposed. This study shows fluctuations in FGM levels in three lemur species likely resulting from changes in the structure of a group with relocation of individuals, construction work in the immediate vicinity of animals or placing another animal in quarters adjacent to the observed individuals. The results suggest that some lemur species may be more or less sensitive to such disturbances. However, FGM concentrations had a similar pattern  during 30 days of monitoring in all three species of


For a large population of elderly who live alone, a continuous long-term daily monitoring system is critical and imminently needed to enhance the quality of their lives. Continuous monitoring of vitality information  is essential for elderly care. In this paper, we use existing commodity Wi-Fi devices to design and implement a long-term device-free human daily vitality system, WiMonitor. Our system can continuously capture the target’s vitality information in a multi-room home environment without compromising the privacy of the target. In a continuous 22-day experiment, WiMonitor successfully captures the human vitality information accurately. We believe our system can provide valuable long-term monitoring data for both researchers and health care personnel. Population aging is a global issue affecting most countries. In 2019, there were 703 million persons aged 65 years or over in the global population, and this number is projected to double to 1.5 billion in 2050 ,5.Elderly people who live alone can develop poor health, they may have an accident at home and have no way of alerting someone that they need help . Thus, aHowever, capturing human daily life information for a long period of time is challenging. An obvious solution is deploying cameras in each room to monitor all the areas and record all activities an elder performs ,12. HoweWi-Fi is particularly promising as Wi-Fi access points and devices are already ubiquitous and thus there is no need of deploying any extra infrastructure. The latest research explores the possibility of employing Wi-Fi channel state information (CSI) for indoor localization ,20,21,22In this paper, we introduce the notion of vitality to represent the statistical measure of one’s staying area (room-level), motion state and activity intensity levels during a specific time period (such as 24 h). Through statistical analysis of vitality information, we can reveal the daily life routine of the elder and find any abnormalities. For example, if the elder stays too long in the toilet in the still state which is different from his/her usual daily life routine, then that may indicate an abnormality calling for attention . There has been prior research work on human vitality sensing—WiVit system . HoweverWe propose a method to achieve robust long-term room-level area detection.We design a metric to estimate the intensity of physical activity and explain the metric’s capability for activity intensity estimation.We implement and deploy a real-time Human Vitality Monitor System—WiMonitor, using commodity WiFi devices for continuous long-term monitoring in a smart home environment, and ask four volunteers to live in the smart home for 24 h in a continuous 22-day experiment. The long-term experiment demonstrates that WiMonitor is able to accurately reveal the daily life routines of different volunteers. The experiment shows that the system can be used to infer the routine habits of people, detect any abnormality. We believe this non-intrusive sensing system and sensing data generated make long-term elderly care and health support at home viable.The main contributions of this paper are as follows:The rest of this paper is organized as follows. In recent years, with the rapid development of sensing technology in the era of the Internet of Things (IOT), a variety of human sensing technologies have been developed, including wearable sensors ,14,32, cThe existing device-free indoor localization systems can be categorized as fingerprint-based and geometric mapping-based. Dang et al.  uses theIn a typical home environment, room-level localization is sufficient for a lot of real-life scenarios. Obtaining the accurate area information  is much more useful than knowing inaccurate fine-grained location information for elder daily life monitoring. WiVit  applies As human motion detection plays an important role in many smart home applications such as intrusion detection and activity recognition, a lot of device-free human motion detection systems have been developed using WiFi CSI. Dong et al.  extracteIn this section, we first introduce the CSI Primer, and then introduce two novel technologies which can reduce the noises of raw CSI signals.h(τ) can be denoted as:ia, iθ and τi represent the amplitude, phase and time delay of the ith path, respectively. N is the total number of multi-path, and δ(τ) is the Dirac delta function [X and Y represent the frequency domain representations of transmitted and received signals, respectively. CFR for a carrier frequency f at time t. H can be represented as:N represents the Gaussian noise [iα is the complex attenuation, id(t) is the propagation length of the ith path, λ is the wavelength and L is the total number of multi-paths.In the indoor environment, WiFi signals propagate from transmitter to receiver through multiple paths, i.e., one Line-of-Sight (LOS) path, and multiple paths from surrounding objects such as walls, furniture and the human body. The signal arriving at the receiver is a superposition of signals from all the paths, this phenomenon is called multipath effect ,38. The function . Channelan noise . In WiFisH is the static component which is the sum of CFRs for static paths, and S is the attenuation, φ is the phase shift of the static component. dH is the dynamic component which is the sum of CFRs for dynamic paths, and kα is the complex attenuation, kd(t) is the path length of the kth path, respectively. The multiple paths can be divided into static paths and dynamic paths . The staAGC and ε. Since the transmitter and receiver are not time-synchronized, there is a time-varying random phase offset However, the CSI measurements provided by commercial WiFi devices contain two major types of noise: the amplitude noise and the phase noise . Due to I sample ,18. ConsThis CSI noise prevents us from achieving satisfactory sensing performance. In order to remove such noise, several signal processing technologies have been proposed. Among these technologies, CSI conjugate multiplication and CSI ratio are two novel technologies which have been widely used in existing research works ,28,40,41A commodity WiFi card usually has multiple antennas, the time-varying random phase offsets are the same across different antennas on a WiFi card as they share the same RF oscillator . Thus, ccmH is the output of conjugate multiplication, The conjugate multiplication between the CSI of two antennas can be denoted as Equation (6), where In Equation (6), the term ① is the product of the static components and can be treated as a constant within a short time period. The term ② and term ③ are two products of static component of one antenna and dynamic component of another antenna. The term ④ is the product of dynamic components which can be ignored as the value is very small. From Equation (6), we can see that the random phase offset has been removed from CSI. However, conjugate multiplication between the CSI of two antennas amplifies the amplitude noise. a = α1, b = c =d = z =Assuming there is only one reflection path corresponding to the human target’s movement, the dynamic component is the path reflected from the human target while the static component is composed of the LoS propagation and other reflection paths from static objects in the environment. The CSI Ratio is calculated by taking the division operation of CSI between two antennas, which is defined as follows:a, b, c, d are all constant complex numbers, z represents a circle in complex plane, its phase represents the dynamic reflected path. Thus, the CSI Ratio can be regarded as the form of Mobius transformation [Since the parameters ormation .CSI Ratio has higher signal-to-noise-ratio(SNR) than raw CSI and CSI conjugate multiplication, and changes following a circular pattern in complex plan;If the reflection path length change is exactly one wavelength, the CSI ratio will rotate exactly 2π in complex plane;If the reflection path length increases, CSI ratio rotates clockwise, otherwise, it rotates counterclockwise.The result of CSI Ratio is still a complex number with amplitude and phase, the amplitude is the ratio of the raw CSI amplitudes and the phase is the phase difference of the raw CSI phases. It has been proved in ,41 that Since WiMonitor is a long-term human vitality monitoring system, it contains two major components: long-term area detection and long-term activity intensity estimation (including motion detection). As for area detection, Wiborder  construcy(t) through a mixer. Due to the path loss and multipath fading, the received signal strength is weaker than the transmitted signal strength. In order to maintain stable received signal strength for better wireless communication, AGC could amplify the signal strength dynamically according to Received Signal Strength Indicator (RSSI). The amplifier gain is large when the received signal strength is weak, so the amplifier gain can be considered as the function of RSSI gy(t) represents the received signal after AGC, gy(t) can be denoted as:gY can be denoted as:Y is the frequency domain of received signal before AGC. According to Equation (2), the CSI after the AGC gain gH can be denoted as:According to the properties of discrete Fourier transfer (DFT), the frequency domain of received signal after AGC δ(RSSI) is the noise which needs to be removed. However, the value of function δ(RSSI) is not accessible due to the hardware limitations of commodity WiFi card. Fortunately, through Intel 5300 CSI Tools [agc is related to δ(RSSI) and is controlled by RSSI, we can use the function γ(agc) instead of δ(RSSI) to remove the noise of AGC. As for wireless sensing, the CSI is modelled to describe the state of wireless channel, which is not include AGC, so SI Tools , we can gy(n) and y(n) be the sampled versions of gy(t) and y(t), respectively. According to Parseval’s Theorem, we can get Equations (12) and (13): N is the number of samples. According to Equations (10), (12) and (13), we can get Equation (14):Let The denominator of Equation (14) represents the received signal strength before AGC. Through Intel 5300 CSI Tools, the denominator can be calculated as:LTF is the Long Training Field, which is used to estimate the CSI [γ(agc)2 can be denoted as:The numerator of Equation (14) represents the signal power after AGC in the frequency domain, and can be calculated as :(16)∑k=0 the CSI . Accordiγ2(agc) and agc, we have collected a sufficiently large volume of CSI data. During data collection, we change the positions of transceivers as much as possible in order to get as many agc values as possible. Then, through Equation (17), we can obtain the value of γ2(agc) corresponding to each agc, and we can get the relationship of γ2(agc) and agc, which is shown in γ2(agc) increases with the increases of agc. That is to say, when the received signal power is small, In order to get the relationship of Thus, we can use Equation (18) to remove the noise of AGC:Due to the noise of AGC, the CSI amplitude could fluctuate when there is no human motion or environment changes. So the thresholds for area detection of Wiborder will change even in the silence environment, thus Wiborder is not suitable for long-term area detection. In order to evaluate the effectiveness of our proposed method for remove the noise of AGC, we collect CSI data for a long time (about 1.5 h) in the silence environment. agc and γ(agc), respectively. There is also has a similar but smaller jump at around 3500 s in the agc of the corresponding CSI amplitude remains relative stable except for some outliers. These outliers may be caused by environmental noise, how to remove the environment noise is out of scope of our paper. From agc and γ(agc) are also divided into two segments at about 5000 s. Therefore, we can conclude that the jumping of amplitude at about 5000 s is caused by AGC. As the Rayleigh fitting parameter is obtained from the amplitude of conjugate multiplication between the CSI of two antennas, when the amplitude contains the noise of AGC, the parameter is not robust to perform long-term area detection.The amplitude of CSI and the conjugate multiplication between the CSI of two antennas after removing the noise of AGC is shown in Since we can get more stable CSI amplitude after remove the noise of AGC, we can obtain more robust Rayleigh fitting parameter compared with Wiborder. In this subsection, we introduce the method for the CSI metric of activity intensity, which include how to extract Doppler frequency from the phase changes of CSI Ratio, and how to estimate activity intensity coefficient. At last, we combine Doppler frequency and activity intensity coefficient to construct the CSI metric for activity intensity estimation.f is the carrier frequency of the signal, pathν is the speed of path length change, and c is the speed of light.In a typical environment with a pair of WiFi transmitter and receiver, as illustrated in t0 is A0 is the attenuation, ϕ0 is the phase of dynamic path at t0, and τ0 is the propagation delay. If the path length changes at a speed of pathν, after the short time period t, the path length change Δpathl = pathtν, and the propagation delay change is ϕ1 is the phase of dynamic path at t1 The phase change Δϕ of dynamic path can be denoted as:Consider only one signal, its CSI at time The Doppler frequency shift in the received signal can be rewritten as:In the real environment, due to multipath-effect, the received signal is a superimposition signals from all paths. We can use Equation (22) to approximatively estimate Doppler frequency which introduced by human movements. t (such as 0.05 s) to calculate the tangent vector π phase jumps. Once we obtain the phase change Δϕ, we can estimate Doppler frequency approximatively using Equation (22). We employ an example to illustrate the Doppler frequency which obtained by this method. We let a person walk towards the ligature of the pair of transceivers and then to walk away twice. In theory, when the person walks towards the ligature of the pair of transceivers, the movement of the person will introduce negative Doppler frequency due to the change in radial component of the motion with respect to the receiver, and vice versa. The extracted Doppler frequency is shown in From Equation (22), we can see that if we can measure the phase change of dynamic path, we can estimate the Doppler frequency. However, due to the random phase offset of raw CSI, the phase change cannot be directly measured from raw CSI data. Since CSI ratio can remove both amplitude noises and phase noises of raw CSI, and has higher SNR than raw CSI, and the phase change of CSI Ratio represents the path length change of reflected path. we can use CSI Ratio to obtain the phase change of dynamic path. In the complex plane, we use The tangent vector corresponding to the dynamic component is a complex vector, and its amplitude represents the distances between consecutive samples. In order to find the physical meaning of the sample distances, we conducted an experiment, the experimental setup is shown as We let a person perform in-place activities at first, and then wave his arm while standing still. M complex data samples within a short time window (such as 2 s), and the tangent vector of dynamic component also has M complex data samples. t0, where Δt1 = 0. We can construct the sample distance vector from tangent vector:α(t0) is the amplitude of tangent vector data sample at time t0.Suppose the CSI Ratio has α(t0 + Δkt) represent the kth element of vector α(t0 + Δkt), we have collected 2 s tangent vector data samples generated by a silent environment and a dynamic environment (where a human performs in-situ activities), respectively. α(t0 + Δkt)) and α(t0 + Δkt), respectively. From α(t0 + Δkt) approximately follows a normal distribution, so the distribution of α(t0 + Δkt) approximately follows a log-normal distribution in both silence environment and dynamic environment.Let μ and σ are the mean and standard deviation, respectively, of the associated normal distribution.The PDF of log-normal distribution is given by:The geometric mean of the log-normal distribution is given by :(25)GM=eη using Equation (26):We construct the activity intensity coefficient η:η is very small;In the silent environment, the value of η is larger when activities are produced by the torso than for activities produced by the arms and legs.Based on Equation (26), we can obtain the following two properties of thresholdf to discriminate human motion and still states. Suppose we have obtained N samples of Doppler frequency sequence N samples of coefficient of activity intensity sequence t (0.4 s), the fraction of time when the user is in the motion state can be denoted as:ξ can be constructed as:Since we have obtained the Doppler frequency and the coefficient of activity intensity, we can construct the CSI Metric for activity intensity estimation which combined with Doppler frequency and the coefficient of activity intensity. When human is still, due to the environmental noises, small Doppler frequency shifts have been observed as phase changes in CSI Ratio, and hence we can use an appropriate threshold In order to evaluate the effectiveness of the proposed metric, we conducted an experiment, and the experiment setup as shown in N is the total number of data samples.The mean of the magnitude of data can be denoted as:As subtracting the mean of the magnitude data can remove any constant effects such as gravity , we subtmagNoG is used as the ground-truth of intensity, magNoG and the CSI metric for these two actions is shown in In this section, we analyse the 22 days of continuous 24-h human vitality monitoring data of four volunteers which recorded by a WiMonitor system. We employ miniPCs equipped with off-the-shelf Intel 5300 Wi-Fi cards as the transmitter and receivers. Each receiver is equipped with two antennas. The CSI tool developed by Halperin  is instaWe have implemented WiMonitor as a real-time system, and ask four volunteers to conduct the experiments. The basic information of these volunteers is shown in The basic information of vitality contains the area detection results, the motion status and activity intensity. In our experiments, at least one Wi-Fi receiver is employed in each subspace. We can choose the receiver in which the target is located in to obtain the activity intensity. For example, as illustrated in As for area detection, we proposed a method to reduce the noise of AGC, thus can provide stable sensing boundary parameter. Based on the area detection method of Wiborder, we can achieve long-term area detection. In order to evaluate the performance of area detection of WiMonitor, we use two commonly used metrics precision and recall to show the performance. These two metrics are calculated by true positive (TP), false positive (FP), true negative (TN), and false negative (FN). For a certain area We calculated the precision and recall for each area separately, and then take the average over all areas. As for motion detection, WiVit  utilizedA continuous 24 h of basic information of human vitality  is shown in Compared with As the WiMonitor system can continuously monitor and record human vitality information in real time, we can analyze the overall activity level of the elderly during one day or even longer to mine the daily living habits and find abnormity.Since WiMonitor system recorded 22 days of continuous 24 h of human vitality information data from four volunteers, in this subsection, we can analyze these data over 24-h from different perspective to capture the living habit patterns of different volunteers. It is well known that in the smart home environment, physical activities are area-related, such as sleeping usually takes place in the bedroom, and cooking usually takes place in the kitchen. Analyzing the area occupation rate during 24 h of the elder, could help us to find out the living habits of the elder. Considering different volunteers may have different living habits in the perspective of long-term area staying information, in order to find whether WiMonitor could capture the living habits of one user and the diversity of habits of different users, we can analyze the accumulated hours of area staying of all the volunteers over 22 days, as shown in Since activity intensity is more fine-grained sensing than motion, in this subsection, we only analyze activity intensity instead. As described in In order to obtain the living habits of one user and the diversity habits of different users in the perspective of long-term activity level information, we can also analyze the activity level rate of all the 22 days of four volunteers in a specific area and in all the four areas overall. Through analysis statistical properties of human vitality information, we not only can capture the living habits of users, but also detect abnormality. Take sleep as an example, since sleep plays an important role in our health and well-being. Low quality of sleep will increase the risk of sleep disorders. In this paper, we can use instantaneous activity intensity to analysis body movements during sleep. We consider motion states as those instantaneous activity intensity values which are larger than zero. For an individual subject, it is true that the body movement during sleep over one night cannot tell if the subject sleeps well or not, however, the body movement pattern over weeks or months could be a good indicator for one’s sleep quality . In this paper, we used the accumulated body movement time during sleep to represent the body movement frequency. In recent years, many device-free indoor localization systems have been proposed, and the existing device-free indoor localization systems can be categorized as fingerprint-based and geometric mapping-based. Many methodologies have been proposed to improve the performance, such as deep learning and transfer learning ,49, etc.precision and recall of our approach are both higher than 96%).Though the above two types of localization approaches are very useful in many applications, such as indoor navigation, augmented reality, disaster rescue, etc. However, in a typical home environment, the rooms are usually divided by walls, detecting which room a person is staying is essential for daily life monitoring. This requires the system to detect the precise sensing boundary between different rooms. However, the above two types of localization approaches couldn’t fulfill this requirement, because 20 cm localization error could make wrong judgement of room-level localization when a person is close to the room door. Different from the above works, our proposed approach uses a novel CSI metric for through-wall discrimination to determine the precise sensing boundary between different rooms, thus can achieve accurate room-level detection (the According to , on averIn this paper, we propose a continuous long-term human vitality sensing system called WiMonitor using commodity Wi-Fi devices. For an elder who lives alone, the WiMonitor system could capture vitality information continuously in real-time without any human effort on offline-training. WiMonitor could achieve continuous long-term accurate area detection by removing the noise of AGC. Moreover, we construct a CSI metric for estimate the instantaneous activity intensity real time. We recruit 4 volunteers to conduct a continuous 22-days experiment, and record the long-term vitality data. Through analysis, we can conclude that the recorded long-term vitality information can be used to obtain the routine habits of the user and detect any abnormality. As the next step, we are going to use our system to track vitality information over a longer term, with the hope that we could observe the long-term routine change trends using the data from several months. We envision our WiMonitor system could be a useful system to provide abundant datasets of the daily living of the elderly to help not just researchers but also healthcare personnel.


We assessed the efficacy of buprenorphine replacement taper therapy in a psychiatric hospital in Japan.Based on the medical records, a retrospective analysis was performed to evaluate the outcomes of buprenorphine replacement taper therapy in 106 subjects with heroin dependence.We found that replacement and taper therapy with buprenorphine could significantly reduce withdrawal symptoms during detoxification. In addition, the completion rate of detoxification was significantly improved and the length of hospital stay was significantly reduced relative to those who received conventional treatment without buprenorphine. However, the readmission rate increased after the introduction of detoxication therapy with buprenorphine.The present findings suggest not only the efficacy and safety of buprenorphine replacement and taper therapy, but also the requirement for maintenance therapy for individuals with heroin dependence.  In addition to its powerful euphoria, heroin induces both psychological and physical dependence, and thus, it is one of the most commonly used illicit drugs worldwide. In the United States, since the 1990s, the widespread use of opioids for noncancer pain has led to a vicious cycle of abuse and resale of these drugs, as well as the abuse of heroin, which can be obtained cheaply, leading to an opioid epidemicIn Japan, the epidemic began to spread briefly around 1960, but it has rapidly declined due to amendments to the illicit drug control law and the drug banishment campaign in Japan. In 2016, the number of heroin arrests and the number of individuals arrested were three and zero, respectively, and the amount of heroin seized was low at 4.540 g in Japan.The number of heroin addicts admitted at Tarumi Hospital in Kobe, Japan, began to increase around April 1998. Most of them were refugees from Southeast Asian descent. Historically, the first boat people came to Japan in May 1975, following which the first Vietnamese refugees arrived in Japan in 1978. Since the permission to settle in Kobe was granted, various problems encountered by Vietnamese living in Kobe were exposed by the Great Hanshin‐Awaji Earthquake in 1995, which led to the establishment of the nongovernmental organization “Vietnam in Kobe” in 2000. Since the first Vietnamese refugee heroin addict was admitted to the hospital in April 1998, the number has been increasing in hospital care.Heroin addiction is characterized by painful symptoms of withdrawal. Although both the pleasure from drug use and the craving that comes from psychological dependence are strong enough, most of the sustained use of heroin abusers is mainly to avoid experiencing the pain of withdrawal symptoms. As to short‐acting heroin, a wide variety of autonomic symptoms, known as “autonomic storms,” emerge several hours after the last use of the substance. Autonomic symptoms, such as general malaise, slight fever and chills, sweating, goose bumps, tears, coughing, and yawning, begin to appear as well as severe pain, such as myalgia, arthralgia, and bone pain, followed by gastrointestinal symptoms such as diarrhea, nausea, and vomiting, accompanied by insomnia and intense anxiety and agitation. Commonly, most patients would visit the clinic to alleviate pain and to discontinue heroin. The treatment should begin with the discontinuation of heroin use . However, when our hospital started to accept heroin‐dependent patients, we experienced difficulties in the management of their withdrawal period. Patients were often secluded due to psychomotor agitation with distress of withdrawal symptoms, and we only administered fluids for gastrointestinal symptoms and anorexia. With regard to medication, sleeping pills, antipsychotics, and anxiolytics were mainly used as a symptomatic treatment for insomnia and anxiety caused by the pain of withdrawal symptoms, which were not much different from the treatment of psychotic disorders such as schizophrenia and other substance‐related disorders. In other words, medications were expected to improve insomnia, agitation, and restlessness associated with withdrawal symptoms, but they were largely ineffective in reducing the pain of withdrawal symptoms. To overcome this situation, buprenorphine (Lepetan®) treatment for opioid detoxification was started on a trial basis in 2002. Subsequently, buprenorphine replacement taper therapy (BRTT) was fully introduced in our hospital in 2005.Buprenorphine is a high‐potency partial agonist that acts on the μ‐opioid receptor. It is currently marketed in Japan as an injection and a suppository for postoperative pain, cancer pain, and myocardial infarction. In brief, buprenorphine is characterized by a high affinity for μ‐opioid receptors, a long blood half‐life, and low intrinsic activity.In the present study, we examined the medical records of 106 heroin‐dependent patients who had been treated with buprenorphine to determine the effect of BRTT by comparing 26 patients who were admitted to the hospital without the administration of buprenorphine for the detoxification of heroin.22.1Establish an initial tentative daily dose of buprenorphine. We administered four ampules of 0.2 mg (0.8 mg) intramuscularly per day, as a start, to patients with standard heroin use. However, if we suspected that the patient could be highly tolerant based on the interview and treatment history, we administered six to eight ampules (1.2‐1.6 mg) per day. Conversely, three ampules (0.6 mg) injection of buprenorphine were initiated in cases with apparently low heroin use.Many patients with heroin dependence usually use heroin until just before admission or until the morning after admission. Because of the persistent effects of heroin at the time of admission, we need to pay attention to the timing of administration, as buprenorphine may induce withdrawal symptoms. After admission, the patient should be monitored for some time while waiting for the onset of withdrawal symptoms. A tentative daily dose of buprenorphine will be administered starting from the time withdrawal symptoms are confirmed. At this point, the acute symptoms of buprenorphine intoxication should not be overlooked . Therefore, the drug should be administered in dose at intervals of 1‐3 hours and discontinued if signs of acute intoxication are observed while confirming safety. If the first day's dose is six ampules or more, two ampules should be administered at a time. Buprenorphine treatment was usually initiated in the late afternoon, but withdrawal symptoms are often insignificant at this point. In cases with high tolerance, withdrawal symptoms may become more severe late at night on the first day after admission, so additional doses may be administered as necessary while the patient is repeatedly examined to monitor the withdrawal symptoms.On the morning of the second day in the hospital, withdrawal symptoms were assessed and the daily dose of buprenorphine was adjusted. If few withdrawal symptoms were observed, the dose of buprenorphine could be reduced from the day 1 buprenorphine dose and continued through day 4. If withdrawal symptoms were persistent but can be objectively and subjectively tolerated, the day 1 buprenorphine dose was maintained, administered in divided doses from morning to lights‐out, and that dose is continued through day 4. In cases where additional doses were administered on the first night or in case severe withdrawal symptoms emerged, such as restlessness, or patients heavily suffering from distress due to withdrawal symptoms, the dose of buprenorphine was increased from the day 1 buprenorphine dosage and additional doses were administered. While monitoring the withdrawal symptoms frequently, one to two ampules of buprenorphine (0.2‐0.4 mg) should be administered at 1‐ to 2‐hour intervals, with no upper limit for additional doses, until withdrawal symptoms improve. Withdrawal symptoms usually improve dramatically once the required dose is met. If additional doses are required, a higher dose is administered, and the dose is continued until day 4. In general, such a treatment course can largely eliminate withdrawal distress on the second day of hospitalization; then, withdrawal symptoms are significantly reduced by the third day (48‐72 hours later), which is regarded as the peak of withdrawal symptoms.Starting on day 5, buprenorphine was gradually tapered off, one ampule every 2 days if the continuous dose by day 4 was low or one ampule per day if the continuous dose was high in the case with high tolerance. Withdrawal symptoms with buprenorphine may occur when buprenorphine is tapered at a faster pace. However, such withdrawal symptoms are much milder and less unbearable than those of heroin withdrawal. Taper off as slowly as possible is recommended, but people who are administered buprenorphine may seek a prompt reduction or eager to leave the hospital during the tapering off. The "endpoint (completion)" of BRTT was fixed as 2 days after the last administration of buprenorphine.Withdrawal symptoms were assessed using the Clinical Opiate Withdrawal Scale (COWS).2.2Based on the currently available medical records, we examined a total of 132 patients with heroin dependence who were admitted to our hospital for heroin detoxification between September 1, 1998, and July 31, 2020. To determine the efficacy of BRTT, those who had comorbid psychiatric disorders were excluded from the present study. Because this was a retrospective observational study based on medical records, informed consent was obtained from the subjects by means of opt‐out consent. Patient information was kept confidential and anonymous. The study and protocols were approved by the Ethics Committee of Tarumi Hospital and were in accordance with the Declaration of Helsinki.The characteristics of the subjects—such as gender, age, nationality, and status of medical insurance—were obtained from their medical records. Factors such as length of hospital stay, duration of buprenorphine administration (days), maximum daily buprenorphine dose (mg), and total buprenorphine dose (mg) were extracted to evaluate the efficacy of BRTT. Furthermore, to assess the severity of withdrawal symptoms, the COWS score at the time of admission, the maximum of the withdrawal symptoms (maximum), and discharge were investigated for 85 subjects who could be confidently tracked through the medical records.2.3We used analysis of covariance (ANCOVA) to test the efficacy of BRTT on the length of hospital stay. The model used treatment group (BRTT vs non‐BRTT) as the main effect, with gender, age, history of hospitalization, nationality, and status of medical insurance as covariates. The non‐BRTT group comprised those who received any conventional treatment before the implementation of buprenorphine use. For the outcome measures, a log‐rank test was conducted to evaluate the completion (retention) rate in the treatment of detoxification between the two; “patient dropout” was considered when patients censored the detoxification treatment for their own reasons. In addition, the influences of potential confounding factors on the completion rate, buprenorphine dose (mg), and the COWS score were assessed using chi‐square test or one‐way analysis of variance, for each variable . Finally, Pearson's correlation analysis was performed to determine the relationship between the COWS and buprenorphine administration. Statistical analysis was performed using SPSS ver. 24.0 for Windows (SPSS Inc).3P = .0021). None of confounding factors affected the length of hospital stay. Furthermore, the completion rate of detoxification treatment was 84% in the BRTT group, which was significantly higher  than in the non‐BRTT group (38%).The demographic characteristics of subjects with heroin dependence are shown in Table r = .37, P < .001), whereas there was no association between the maximum scores of COWS and the length of hospital stay .The means of buprenorphine dosage, its dosing duration (days), and the COWS scores during detoxification with BRTT are summarized in Table y Figure . Finally4To the best of our knowledge, the present study is the first report investigating the efficacy of BRTT in people with heroin dependence in Japan.r = 0.37) implies that buprenorphine dosage could be adjusted according to the situation by frequent interviews and observation for withdrawal symptoms, rather than by the COWS scores alone in clinical practice. There was no correlation between the maximum COWS scores and length of hospital stay, suggesting that stronger symptoms during the withdrawal phase did not necessarily lead to prolonged hospitalization. Instead, BRTT may have contributed to a shorter hospital stay for heroin addicts requiring detoxification.The length of hospital stay was significantly shorter for patients who received BRTT, and BRTT improved the completion rate of heroin detoxification, as compared to the non‐BRTT group. The current completion rate of treatment (84%) was consistent with previous reportsDetoxification with buprenorphine use is completely different from conventional treatments such as immediate abstinence from heroin. Indeed, the introduction of BRTT had a significant impact on those who treated patients with heroin dependence in our hospital. The medical staff in psychiatric hospitals tend to feel distressed when accepting people who exhibit hostility and aggressive behavior, physical problems, or uncertain outlooks about inpatient care. After BRTT was introduced in our hospital, patients with heroin dependence rarely became agitated due to withdrawal symptoms. Although mild side effects such as constipation and delirium were observed, we could have a better outlook on the process of withdrawal symptoms and can prepare to deal with exacerbations. In addition, linguistic communication was a major problem because the majority of patients were non‐Japanese. However, the introduction of buprenorphine can contribute to a significant reduction in anxiety and distress for medical staff.We found an increase in readmission rates following the introduction of detoxification therapy with buprenorphine. One possible explanation for the higher rates of readmission is that treatment with buprenorphine greatly alleviated the distress of withdrawal symptoms, which may have resulted in patients who were eager to enter the hospital again for detoxification in the case of a relapse without hesitation. Importantly, higher rates of readmission indicated the importance of maintenance therapy. Indeed, treatment for substance abuse disorder should not only be restricted to short‐term detoxification and symptomatic treatment, but also include maintaining long‐term follow‐up with psychosocial approaches such as cognitive behavioral therapy, contingency management, and utilizing mutual aid groups such as narcotics anonymous and other social supports. The goal is to prevent high‐risk behaviors that lead to relapse and promote social adjustment and human maturity. In other words, detoxification should be only the beginning of treatment for people with heroin dependence. In fact, we found that only a few patients attended the hospital after they were discharged in our hospital. Although the introduction of buprenorphine increased the motivation for detoxification treatment, it was extremely difficult to translate into the treatment for long‐term maintenance of abstinence under the current conditions, at least in our Japanese hospital. For this reason, maintenance therapy with opioid receptor full agonists or opioid receptor partial agonists may be desirable in addition to psychosocial interventions when considering the WHO guidelines.Based on a large number of past practices, we demonstrated the efficacy of BRTT in the detoxification of heroin dependence. However, the present study has some potential limitations. First, the BRTT was exclusively performed in our hospital, and the majority of the patients were foreign nationals, all of which may have caused selection bias. Second, all subjects received psychotropic medications at least once during hospitalization, and people who had comorbid other psychiatric disorders were excluded from this study. In addition, factors affecting BRTT treatment, such as heroin dosage, duration of heroin use, developmental history, and socioeconomic factors, were not considered in the present study. Prospective studies should validate the effectiveness of BRTT while considering the extent to which these factors can affect BRTT.Finally, detoxification with buprenorphine is the only medication that can be administered to people with heroin dependence in Japan. Psychosocial interventions play crucial roles in heroin addicts, who are typically chronic and relapsing. However, we sincerely hope that the Japanese law that prohibits maintenance therapy using methadone will be reviewed in the future and that high‐dose buprenorphine sublingual tablets will be introduced as a treatment option for heroin dependence in Japan.The authors have no conflicts of interest to declare.TN, TA, TK, YY, and KO were involved in data collection. TN and S. K. analyzed the data and wrote the manuscript. NY and To.K. supervised the entire project. All authors have read and approved the final manuscript.This study was approved by the Ethics Committee of Tarumi Hospital, Japan.Informed consent was obtained from the subjects by means of opt‐out consent.Table S1Click here for additional data file.


The growing utilization of needle biopsy has challenged the current pathology consensus of IgG4-related disease (IgG4-RD). The aims of this study were to identify the histological characteristics of needle biopsy and surgical specimens and evaluate the ability of needle biopsy in histological diagnosis of IgG4-RD.Biopsies from patients who were referred to as IgG4-RD by the 2019 ACR/EULAR IgG4-RD classification criteria in Peking University People’s Hospital from 2012 to 2019 were re-evaluated. Typical histological features and diagnostic categories were compared between needle biopsy and surgical biopsy.P = 0.033). Surgical biopsy showed significantly higher IgG4+ plasma cells/high-power field (IgG4/HPF) count (P < 0.01) and higher proportion of IgG4/HPF > 10 (P < 0.01). No significant difference was observed with regard to the ratio of IgG4+ plasma cells/IgG+ plasma cells (IgG4/IgG) (P = 0.399), storiform fibrosis (P = 0.739), and obliterative phletibis (P = 0.153). According to the 2011 comprehensive diagnostic criteria, patients who performed a needle biopsy were less likely to be probable IgG4-RD (P = 0.045). Based on the 2011 pathology consensus, needle biopsy was less likely to be diagnosed as IgG4-RD (P < 0.01), especially to be highly suggestive IgG4-RD (P < 0.01). Only 1/18 (5.6%) needle salivary specimens fulfilled the cutoff of IgG4/HPF > 100, which was significantly less than 15/23 (65.2%) of surgical ones (P < 0.01).In total, 69 patients met the 2019 ACR/EULAR classification criteria and 72 biopsies of them were re-evaluated. All cases showed lymphoplasmacytic infiltrate, while storiform fibrosis and obliterative phlebitis were only present in 35 (48.6%) and 23 (31.9%) specimens, respectively. Storiform fibrosis was more likely to be seen in retroperitoneum lesion (Needle biopsy shows an inferiority in detecting IgG4/HPF count but not in IgG4/IgG ratio, storiform fibrosis, and obliterative phlebitis. Compared with surgical samples, needle biopsy is less likely to obtain a histological diagnosis of IgG4-RD. A different IgG4/HPF threshold for needle biopsy of the salivary glands may be considered.The online version contains supplementary material available at 10.1186/s13075-021-02432-y. IgG4-related disease (IgG4-RD) is a newly recognized fibroinflammatory condition characterized by tumefactive lesions involved in multiple sites; often but not always, the elevated serum IgG4 concentration; and a dense lymphoplasmacytic infiltrate rich in IgG4-positive plasma cells . ComprehWe performed a comparison of needle biopsy and surgical biopsy in terms of histological features and diagnostic categories and re-evaluated the 2011 pathology consensus based on 72 biopsied specimens  from 69 patients who underwent biopsy and were referred to as IgG4-RD by the 2019 ACR/EULAR IgG4-RD classification criteria in Peking University People’s Hospital from 2012 to 2019 [Patients who underwent biopsy  in Peking University People’s Hospital and were histopathologically suggested or suspected as IgG4-RD from 2012 to 2019 were enrolled. All patients provided written informed consent. The following data were collected from the medical records: gender, age of disease onset, and baseline serum IgG4 concentration. The disease activity reflected by IgG4-related disease responder index (IgG4-RD RI) was calculated by two independent physicians and mean value was taken .Two pathologists who were blind to the sample information worked independently and re-evaluated the hematoxylin and eosin (H&E)–stained, IgG4 and IgG stained slides of enrolled patients. The degree of fibrosis and lymphoplasmacytic infiltrates was assessed. Typical histopathological characteristics of storiform fibrosis and obliterative phlebitis were recorded. Three 40× fields with the highest number of IgG4 plasma cells were calculated, and the average number was recorded. The same three fields were counted for the IgG4/IgG ratio , 3. In ct test. Categorical variables were assessed with the chi-squared test or Fisher’s exact test, as appropriate. P values were adjusted with Bonferroni method when comparing multiple categorical variables in pairs. P values < 0.05 were considered statistically significant. All statistical analyses were performed by SPSS version 25.0. All figures were made by the Origin 2018.Quantitative variables with non-normally distribution were presented as medians and interquartile range (IQR) and were compared with Mann-Whitney test. Quantitative variables with normally distribution were presented as mean ± standard deviation (SD) and were compared with Student’s This study was approved by the Medical Ethics Committee of Peking University People’s Hospital .In total, 69 patients met the 2019 ACR/EULAR criteria, whose classification criteria scores were all above 20 and the median score was 39.5 . Forty-four of the 69 patients (61.1%) were males, and the median age was 56.0 . The median IgG4-RD RI was 4.0, and the median serum IgG4 concentration was 770 mg/dl. Three patients underwent two biopsies in different organs, and 72 specimens in all were re-evaluated  was the most common biopsy site. Other involved organs included retroperitoneum , lymph nodes , lacrimal gland , lung , pancreas , meninges , pleura , bile duct , and prostate . According to the 2011 comprehensive diagnostic criteria, 11 (15.3%) of the patients were possible, 14 (19.4%) were probable, and 47 (65.3%) were definite IgG4-RD. For patients who performed two biopsies, the diagnostic categories were identical. According to the 2011 pathology consensus, 30 (41.7%) of the biopsies were insufficient, 18 (25.0%) were probable, and 24 (33.3%) were highly suggestive IgG4-RD (Table P = 0.177). Sixty-two (86.1%) and 66 (91.7%) biopsies showed IgG4/IgG > 40% and IgG4/HPF > 10, respectively. Storiform fibrosis and obliterative phlebitis were comparatively uncommon and were only present in 35 (48.6%) and 23 (31.9%) specimens, respectively (Table P = 0.446), while storiform fibrosis was more likely to be seen in retroperitoneum lesion (P = 0.033) , and the average IgG4/IgG ratio was 67.5 ± 23.3%. Retroperitoneum specimen tended to show less IgG4/HPF count than other organs, even though no significant difference was found  demonstrated no difference between the two groups. No significant discrepancy was found between needle biopsy and surgical biopsy in terms of IgG4/IgG ratio (P = 0.438), proportion of IgG4/IgG > 40% (P = 0.399), storiform fibrosis (P = 0.739), and obliterative phlebitis (P = 0.153). However, needle specimens tended to show significantly less IgG4/HPF count (P = 0.003) and lower proportion of IgG4/HPF > 10 (P = 0.001) than needle biopsy. Compatible with that, according to the 2011 comprehensive diagnostic criteria, patients who performed a needle biopsy were less likely to be probable IgG4-RD (P = 0.045) while serum IgG4 showed no significant difference (P = 0.618). Consistently, based on the 2011 pathology consensus, needle biopsy was less likely to be histologically identified as IgG4-RD (P < 0.001), especially to be highly suggestive IgG4-RD (P < 0.001)  specimens were surgical biopsy. IgG4-RD RI (P = 0.939), IgG4/IgG ratio (P = 0.437), prevalence of IgG4/IgG > 40% (P = 1.000), storiform fibrosis (P = 0.711), and obliterative phlebitis (P = 0.191). However, IgG4/HPF count (P < 0.001) and the proportion of IgG4/HPF > 10 (P = 0.008) in surgical biopsy were dramatically higher than that in surgical biopsy (P < 0.001). These data suggested that needle biopsy might be less capable to capture full IgG4 profile and thus insufficient IgG4/HPF count, which led to less possibility of histological identification based on the 2011 pathology consensus statement.Given the limited samples obtained from the meninges, lung, pleura, pancreas, bile duct, and prostate, we excluded these entities for repetitive analysis. Compatible with the results above, no difference was observed pertaining to IgG4-RD RI  were highly suggestive, 18 (25.00%) were probable, and 30 (41.67%) were insufficient to be identified as IgG4-RD. Given that all the 72 specimens showed at least one of the three pathological features (lymphoplasmacytic infiltrate), all of these insufficient biopsies failed to fulfill either required IgG4/HPF cutoff point or IgG4/IgG > 40%. Even though IgG4/HPF > 10 had been proposed as one component of a comprehensive diagnostic panel, the pathology criteria recommended a set of IgG4/HPF threshold that was specific to each organ, from 10 to 200. In the 30 (41.67%) insufficient IgG4-RD specimens, 7 (9.72%) showed IgG4/IgG ≤ 40% and 29 (40.28%) showed less IgG4/HPF count than the cutoff value as recommended by the consensus (Table In the 29 (40.28%) specimens with insufficient IgG4/HPF count, 23 (31.94%) were taken from salivary glands, 2 (2.78%) were from lymph nodes, and 1 (1.39%) was from the lung, lacrimal gland, pleura, and pancreas, respectively. The cutoff IgG4/HPF value of the salivary gland recommended by the pathology consensus was 100/HPF. Nevertheless, in this cohort, only 16 out of 41 (39.02%) salivary samples had met this threshold, and 15 of them were surgical specimens while only 1 was needle biopsy (Table P = 0.695), IgG4/IgG (P = 0.720), and proportion of IgG4/IgG > 40% (P = 1.000). But surgical biopsy tended to show significantly higher IgG4/HPF count (P < 0.001) in salivary samples. No more than 1/18 (5.6%) salivary needle specimens met the cutoff value of IgG4/HPF > 100, which was significantly less than 15/23 (65.2%) of surgical ones (P < 0.001). Additionally, salivary needle biopsy was more likely to be neither highly suggestive nor probable IgG4-RD (P < 0.001). These data suggested that, compared with surgical biopsy, needle biopsy was hardly able to capture required IgG4/HPF count and thus be judged as IgG4-RD in salivary glands based on the pathology consensus  were endorsed for the histopathological judgment based on the three histological features , and immunohistochemical features (ratio of IgG4/IgG and IgG4/HPF count). In our cohort, nearly half of salivary specimens were defect of enough IgG4/HPF count for histological identification. Similarly, Andrew et al. reported that two thirds of lacrimal IgG4-RD cases failed to be diagnosed using a cutoff of IgG4/HPF > 100 as recommended by the consensus, but the difference between needle biopsy and surgical samples was not explored , 12, in Despite the superiority of surgical biopsy in detecting IgG4/HPF count, IgG4/IgG ratio showed no divergence between two groups. Even though IgG4/IgG ratio was deemed as a more powerful tool than IgG4/HPF in establishing the diagnosis of IgG4-RD; however, in the absence of other corroborative findings, we are unable to accept IgG4/IgG > 40% itself as sufficient diagnostic evidence in the light of the pathology consensus . This apThe major issue of needle biopsy is represented by the inadequacy of the material obtained for histopathological evaluation and immunohistochemical tests, and thus incomplete characterization of a lymphoproliferative disorder , 15. BesHowever, needle biopsy still has some advantages. Compared with surgical resection, needle biopsy is less invasive, less expensive, and usually with fewer long-term or transient complications, which have a high impact on the patient’s acceptance, especially when the biopsy of an internal viscera is needed. Moreover, even though open surgical biopsies allowed the adequate material for pathological evaluation, it is a non-targeted approach. Regions with restricted lesions may not represent the full features of the disorder. In contrast, ultrasound-guided needle biopsy is able to distinguish areas with different sonographic patterns and target the most suspicious lesions . In addiImproving the specificity of the pathology criteria by setting high-organ-specific cutoffs for IgG4 staining is essential. Even though we interpreted a lower IgG4 cutoff for needle biopsy on salivary gland, it remains to figure out whether needle biopsy is feasible or not for the diagnosis of IgG4-RD and if a different IgG4/HPF cutoff for needle biopsy might impair the specificity. Last but not least, despite the importance of pathology, additional clinical, serological, and radiological evidence is still indispensable for confirming the ultimate diagnosis of IgG4-RD. Patients who lacked one or more of the histological and immunohistochemical features of IgG4-RD may overlap with those showing definite features with regard to serum IgG4 levels, multiorgan involvement, and response to glucocorticoid therapy . Even caWhat should be emphasized is that, biopsies from the same organ of the same patient could be more representative. However, biopsy method was determined by the entity site and patients’ acceptability. Matched samples of both needle and surgical biopsy from the same patient were hardly to obtain. Only one patient in our cohort had matched specimens, whose surgical biopsy also showed slightly higher IgG4/HPF count. In spite of that, demographic characteristics and disease activity reflected by IgG4-RD RI demonstrated no significant disparity, suggesting the comparability of the two biopsy method groups and the reliability of our findings.One limitation is that, the cohort of this study does not cover all the common lesions of IgG4-RD, and most of the samples were taken from the salivary glands. It is still unable to validate the diagnostic capability of needle biopsy with regard to calculating IgG4/HPF count when dealing with other organ samples. It remains to figure out whether a different diagnostic threshold of IgG4/HPF for other entities should be set.In conclusion, needle biopsy shows an inferiority in detecting IgG4/HPF count but not in IgG4/IgG ratio, storiform fibrosis, and obliterative phlebitis. Compared with surgical samples, needle biopsy is less likely to obtain a histological diagnosis of IgG4-RD. A different IgG4/HPF threshold for needle biopsy of the salivary glands may be considered.Additional file 1:. Supplementary Table 1. 2011 Comprehensive clinical diagnostic criteria. Supplementary Table 2. 2011 pathology consensus. Supplementary Table 3. Histological Characteristics in 4 biopsy organs of classified as IgG4-RD. Supplementary Table 4. Comparison of Needle Biopsy and Surgical Biopsy in 72 diagnosed IgG4-RD samples.


Monotonic push-out tests were carried out on 11 specimens having high-strength bolt, T-type perforated plate, or slot-type perforated plate connections to investigate the influence of different connection types on the interface performance of glass fiber reinforced polymer (GFRP)–concrete composite beams. The effects of the number of rows and spacing of high-strength bolts on the failure mode, load–slip relationship, and shear capacity were analyzed. The effects of the number and spacing of holes in the perforated plates, and the inclusion of transverse rebar were analyzed. The results show that the failure mode of the bolt specimens is brittle failure and the perforated plate is ductile failure. The single bolt connection has the lowest ultimate bearing capacity, while the single T-shaped and slotted perforated plates are 170% and 270% times greater. The rows and spacing have no difference in bolts. The perforated plate gradually decreased with an increase in rows and gradually increased with an increase in spacing. The transverse rebar can increase the ultimate bearing capacity and ductility in the plastic stage for perforated plate. Accordingly, there are good choices for interface connectors for GFRP–concrete composite beams, while bolt connectors need to be carefully chosen. Fiber reinforced composite (FRP) is a new type of material with high performance, which is composed of fiber material and matrix material. Glass fiber reinforced polymer (GFRP) refers to the resin matrix that is reinforced by glass fiber. Fiber-reinforced polymer (FRP)–concrete composite beams are a new type of structure that can combine the respective advantages of the two materials, including concrete’s compressive strength and FRP’s high tensile strength . There aOver the last two decades, many studies have investigated the connections between FRP and concrete. Canning et al.  carried In general, experimental studies on shear connectors in FRP–concrete composite structures are still lacking and they have largely focused on the cementation and bolt connection, while research on perforated plate specimens is still in its infancy. Accordingly, limits the use of FRP–concrete composite structures. In this study, an experimental investigation of the shear behavior between GFRP and concrete was carried out. The interconnection slip behavior of the steel bolts and perforated plates is compared based on the load–slip curves, shear capacity, and shear stiffness.The push-out test method is used in this study because there is no standardized method to test FRP shear connectors. A total of 11 specimens were designed in accordance with Eurocode 4 , includiThe mechanical properties of the concrete were determined in accordance with the relevant standard for the test method for the mechanical properties of ordinary concrete (GB/T 50081-2002) .First, the GFRP surface was sanded and the contact surface was rinsed with acetone. Subsequently, the matched glue solution was applied on both sides of the surface of the test piece. After applying the glue, but before the glue had solidified, a torque wrench was used to tighten the connecting bolts. All of the push-out tests were performed while using a force-controlled 500 t loading apparatus . A steelThe strains near the main GFRP beam, holes in the GFRP, bolts, and transverse rebar were measured while using electrical resistance gauges during the loading process, as shown in The failure modes of the four high-strength bolts are all shear failure after the concrete is crushed based on the test results. Specimen B3-R2-S100 is taken as an example to illustrate this failure mode When the load reaches 40 kN, the load decreases slightly. Before reaching the ultimate load, the load increases linearly, and the interface between the GFRP main beam and concrete peels off. When the load reaches 250 kN, the bolt is obviously bent, and the upper end of the gasket is separated. The displacement of the main beam gradually increases, and the upper loading surface exhibits significant compression deformation. Two bolts on one side suddenly fractured at approximately 292 kN. The failure modes of specimens P1-R2-S150 and P2-R3-S100 are shear failures of the concrete tenon and the transverse rebar. Taking P1-R2-S150 as an example to illustrate this failure mode, when the load reaches 240 kN, the GFRP tearing sounds can obviously gradually begin to be heard and, as the load increases, the fracture sound becomes more intense. When the load reaches 245 kN and 265 kN, the first and second cracks appear on the right and left sides of the upper concrete, respectively. Observing the failure mode of the internal structure of the P1-R2-S150 test piece, it can be seen that the concrete tenon and the transverse rebar are severed. The cutting failure of the perfobond GFRP occurs along the direction of the opening line. The GFRP in the failure area is pressed into a white powder form, and the GFRP cracked along the direction of the fiber lines between two pre-drilled holes. The failure mode of specimen P4-R2-S100 with a T-shaped perforated plate is the separation of the interface between the GFRP beam and concrete slab. P4-R2-S100 is taken as an example for illustrating this failure mode. In the beginning, the sound of the GFRP tearing could be clearly heard until the load reached 90 kN. When the load reached 169 kN, the first crack appeared on the left side of the concrete slab and then expanded. When the load reached 234 kN, a second crack appeared on the right side of the concrete slab. The load stopped increasing once it reached 253 kN; when the load decreased to 151 kN, the sound of the reinforcement fracturing could be heard, and the GFRP and concrete were obviously separated. There are two failure modes for the slotted perforated plate, G1-R3-S100 and G2-R2-S150; however, these are similar to the failure mode for the first T-shaped perforated plate and, thus, will not be discussed further here. Another failure mode is the failure of the bond layer between the perforated plate and GFRP beam, which is illustrated by specimen G3-R2-S100. During the test, the sound of a GFRP connector tearing could be clearly heard as a result of shear failure. Finally, the whole concrete part on one side fell off, the bond layer between the opening plate and the GFRP beam was damaged, and the fiber of the GFRP was pulled off. It was found that there was no coarse aggregate in the concrete tenon, and it was sheared together with the transverse rebar during the loading process. The displacement value in the figure is the average value that is recorded by four dial indicators at the lower end of the test component to reduce the influence of the component manufacturing error.When comparing the strains of the main beam with the bolted connection and the GFRP perforated plate connections can reveal the difference in the stress distribution between the bolted connection and the hybrid connection (bonded and bolted). The perforated plate and the GFRP main beam enhance the overall performance of the connection through the hybrid connection. For an interface connector, it is helpful to know the distribution of the interface shear force and the redistribution of stress in the plastic state by elucidating the variation in the stress with the load through strain. When the load is less than 50 kN, the strain measurement point exhibits little change, as shown in It is necessary to ensure that the perforated plate does not undergo brittle fracture during loading. The perforated plate is on the shear plane, and the strain gauge can easily fall off or be damaged during the loading process. Here, the strain–load curve for specimen P1-R2-S150 in the elastic stage is considered. It can be seen from The transverse rebar model number is HRB400, and it has a diameter of 6 mm. When the strain reaches 2105 uP is the experimentally obtained maximum load of the specimens in the push-out tests and n is the number of connectors in the specimen. It stands for the number of high strength bolts and number of openings in the perforated plate respectively.It is assumed that every connector in the same specimen was subject to the same force. Thus, the shear capacity per bolt or concrete wedge can be given, as follows:From In contrast to the bolt specimens, the T-shaped perforated plate is more sensitive to the spacing and number of rows. When the spacing increases from 100 mm to 150 mm, the bearing capacity of a single connection increases by 11.11%. As the spacing increases, the stress diffusion at the opening on the compression side is more uniform and the bearing capacity increases. The bearing capacity decreases by 7.1% when the number of rows increases from two to three. This phenomenon is mainly attributed to the overlapping action zones in the specimens. When comparing the bearing capacities of single connections of specimens P1-R2-S100 and P3-R2-S100-NR reveals that the contribution of the perforated reinforcement to the bearing capacity reaches 43.18%. The transverse rebar plays an important role in the bearing capacity. Generally speaking, the spacing and number of rows affect the bearing capacity of T-shaped perforated plates. The spacing can be increased appropriately and the number of rows can be reduced in order to improve the working efficiency of these connectors.As with the T-type connector, the spacing and number of rows have a significant influence on the bearing capacity of the slot-type perforated plates. When the number of rows increases from two to three, the bearing capacity of a single connection of G1-R3-S100 is 23% lower than that of G3-R2-S100. After increasing the number of rows, the stress superposition between the holes is not conducive to stress diffusion. Comparing G2-R2-S150 and G3-R2-S100 shows that, when the spacing increases from 100 mm to 150 mm, the bearing capacity increases by 13.8%, and the stress diffusion on the compression side of the opening is more uniform. As compared with the T-type perforated plate, the shear area of the slot-type perforated steel bar is greater, and the concrete tenon at the two holes provides shear and, thus, the ultimate bearing capacity of the slot-type perforated plate is approximately 60% higher than that of the T-type perforated plate.uP, and For the composite shear connectors, the shear stiffness is an important index for evaluating the performance of the composite structure, and can be represented by the secant slope of the load–slip curve. A comparison of the shear stiffness will be meaningless unless a uniform equation is used owing to the different spacings of the connectors along the beam direction. The secant stiffness corresponding to 70% of the ultimate bearing capacity is taken as the shear stiffness for the calculation of stud connections in Eurocode 4, as follows:uP, and The Japanese Steel Structure Association  takes thD, is defined as the ratio of the limit slip value The ductility coefficient, In At present, there is no standard and specification for the calculation of the bearing capacity of FRP section concrete composite structure bolt connections. However, the calculation method of bolt connections in steel-concrete composite structure is relatively mature. Here, as a reference, the failure modes of bolt connections are classified into two types: bolt shear failure and concrete failure. In this test, the failure modes of bolt members are bolt shear failure.n is bolt number in one push-out specimen; The current design codes, including those promulgated by AISC , ACI 2424, AASHTThe reduction factor n et al. , Correian et al. . CompariThe structure of the perforated plate connector is similar to that of the PBL connector in the steel-concrete composite structure, and its stress mechanism is similar. The transverse rebar and the concrete tenon share the bearing capacity of the joint, according to the failure mode .Among th(1)N) is the design value of shear capacity of single hole of PBL connector (N); Code for design of steel and concrete composite bridge (GB 50917-2013) (6)Ncv=2(2)N); Specifications for Design and Construction of Highway Steel-concrete Composite Bridge(JTG/T D64-01) (7)Vpu=1In This paper presents the experimental results of the push-out tests on pultruded GFRP I-girders and concrete slabs. Three groups of push-out tests were performed to evaluate the advantages of different connectors for improving the connection between the GFRP girder and concrete slab. The failure modes of specimens with different connection types and the variation in the load–slip curves were analyzed.(1)The failure mode of the bolt specimen is a severing of the bolt bar after the concrete around the bolt is crushed; the specimen undergoes brittle failure. There are two failure modes for the T-shaped perforated plate with transverse rebar: one is the severing of the concrete tenon and transverse rebar, the other is peeling off of the interface between the GFRP beam and the concrete; all of the specimens undergo ductile failure. The failure mode of the T-shaped perforated plate without transverse rebar is a severing of the concrete tenon; the specimen undergoes brittle failure. There are two failure modes for the slot-type perforated plate: one is a severing of the concrete tenon and transverse rebar, the other is damage to the bond layer between the opening plate and GFRP beam; and, the specimens undergo ductile failure.(2)The load–slip curve for the bolt specimen is small at the initial stage of loading; when the ultimate load is reached, the load changes abruptly, and the sliding deformation ability becomes poor. The load–slip curve for the T-shaped perforated plate specimen exhibits an elastic stage and plastic stage, and the plastic stage retains good sliding deformation ability after reaching the ultimate load. The load–slip curve for the slotted perforated plate specimen is the same as that for the T-shaped perforated plate specimen. The difference is that there are two contact surfaces between the perforated steel bar and the opening. After one side is damaged, the other side retains some bearing capacity, which causes the load–slip curve to exhibit a platform for a period after the sudden drop.(3)The ultimate bearing capacity of a single connection is the lowest for the bolt connection; the T-shaped perforated plate is 1.7 times greater and the slot-shaped perforated plate is 2.7 times greater. The ductility coefficient and shear rigidity of the T-shaped perforated plate are the highest, followed by the slotted perforated plate, and the bolt has the lowest values.The following conclusions can be drawn from this study:On the whole, the T-shaped and slotted perforated plates exhibit good anti-sliding ability and a high bearing capacity and, thus, they are good choices for interface connectors for GFRP–concrete composite beams, while bolt connectors need to be carefully chosen.