In order to address the discontinuity caused by the direct application of the infrared and visible image fusion anti-halation method to a video, an efficient night vision anti-halation method based on video fusion is proposed. The designed frame selection based on inter-frame difference determines the optimal cosine angle threshold by analyzing the relation of cosine angle threshold with nonlinear correlation information entropy and de-frame rate. The proposed time-mark-based adaptive motion compensation constructs the same number of interpolation frames as the redundant frames by taking the retained frame number as a time stamp. At the same time, considering the motion vector of two adjacent retained frames as the benchmark, the adaptive weights are constructed according to the interframe differences between the interpolated frame and the last retained frame, then the motion vector of the interpolated frame is estimated. The experimental results show that the proposed frame selection strategy ensures the maximum safe frame removal under the premise of continuous video content at different vehicle speeds in various halation scenes. The frame numbers and playing duration of the fused video are consistent with that of the original video, and the content of the interpolated frame is highly synchronized with that of the corresponding original frames. The average FPS of video fusion in this work is about six times that in the frame-by-frame fusion, which effectively improves the anti-halation processing efficiency of video fusion. Due to the halation phenomenon caused by the abuse of high beam lights at night, drivers approaching from opposite directions are unable to see the road conditions clearly, leading to potential traffic safety hazards .Passive anti-halation methods are simple and effective, such as placing shading boards and growing plants on the middle isolation belts of two-way lanes . HoweverIn comparison with other active anti-halation methods, such as placing polarization film on the front windshield , infrareFrame extraction technology can reduce the redundant frames of video to a certain extent and improve the fusion efficiency. The frame selection method based on sparse representation  is simplIn order to address the problem that the frame rate of the extracted video is inconsistent with that of original video, it is necessary to make interpolation compensation for the retained frames to improve the visual smoothness. The laconic smooth technique based on multi-frame transformation asynchronously (LSTMTA)  is simplIn this work, an efficient anti-halation method suitable for video fusion is presented. Considering the characteristics of small differences in content and high redundancy between adjacent video frames, a frame selection based on inter-frame difference (FSIFD) is designed to minimize the number of fused frames under the premise of continuous video content for improving the fusion efficiency. In addition, a time-mark-based adaptive motion compensation (TMBAMC) is designed for restoring the length of the anti-halation video to be consistent with the original video, and ensuring the content synchronization of corresponding frames. The proposed method is applied to a vehicle head-up display (HUD), which can produce a video without halation, with clear details and rich colors, to assist drivers in driving safely under a special night-time halation scene. In addition, it can be applied to advanced driver assistance systems (ADAS) or autonomous vehicles (AV) to improve the vehicle’s environmental perception ability in special scenarios.The remaining text of the article is arranged as follows. The anti-halation method of infrared and visible image fusion makes use of the complementarity of different source images. The resulting fused image contains minimal halation and rich texture details, as shown in It is notable that there are only small differences in the content between adjacent frames of videos. This results in a lot of redundant operations in achieving the anti-halation image by frame-by-frame fusion, consequently leading to low efficiency. In order to address this issue, the FSIFD strategy is designed by analyzing the motion information of objects between adjacent frames. The strategy ensures that the redundant frames are discarded to the maximum extent in the original video under continuous video content, and only the retained frames are fused by the improved IHS-Curvelet algorithm to meet the high efficiency requirements.In order to solve the problem of the shorter playing duration of the fused video caused by the reduced frames, this work proposes a TMBAMC algorithm to interpolate new frames. Considering the frame number as a time stamp, the interpolated frames with the same time and quantity as the discarded redundant frames are constructed, so that the number of frame and length of the fused video are restored to be consistent with that of the original video. In order to synchronize the content of the interpolated frames with that of the corresponding original frames, we take the motion vector of the two adjacent retained frames as the benchmark, construct the adaptive weights according to the inter-frame difference between the interpolated and the last retained frame, then estimate the motion vector of the interpolated frame.The overall block diagram of the proposed night vision anti-halation method is presented in The difference between two adjacent frames is often measured based on Euclidean distance or cosine similarity. Compared to the Euclidean distance  that caliPV of the ith pixel in a frame is expressed as:iPV ∈ , i ∈ , and L represents the total number of pixels in a frame. iR, iG, and iB represent the intensity of red, green, and blue channels at ith pixel, respectively. ⌊•⌋ denotes the floor operation.The three-dimensional information of a color image is reduced to a one-dimensional array by gray partition. The gray levels in the range of  in each channel are divided into four intervals, including , , , and . Then, the intensity PVin of pixels corresponding to 64 intensity values in a frame is counted to form a feature vector as:The number XYθ of their vectors X and Y:X = [x0 x1 … x63] and Y = [y0 y1 … y63]. The closer XYθ is to 0, the closer cosXYθ is to 1, and the higher the similarity of the two images, the smaller the difference.The difference between two arbitrary frames is expressed as the cosine angle τ to discard the redundant frames in the video sequence. Therefore, the τ directly affects the number of frames removed from a video. If τ is too high, the video will become discontinuous due to too many frames removed, resulting in flickering and skipping. On the contrary, if τ is too small, there will still be redundant frames in the video sequence, leading to poor computational efficiency. Therefore, the threshold τ of cosine angle should meet the requirement of maximum of discarded frames on the premise of continuous video content.The difference between frames is compared with the cosine angle threshold τ with NCIE and DFR, the optimum threshold is determined to satisfy the maximum of discarded frames under the premise of continuous video content.We introduce an objective indicator of overall continuity of a video, namely nonlinear correlation information entropy (NCIE) ,21, and K frames, let the vectors of frames m and w be M = [m0 m1 … m63], W = [w0 w1 … w63]. Then, each data pair  is characterized in an r × c two-dimensional network, where j ∈ , r and c denote the number of rows and columns in the two-dimensional data respectively, where 1 ≤ r = c ≤ 8. The nonlinear correlation coefficient mwNCC between the mth frame and the wth frame is expressed as:rcP denotes the joint probability distribution of M and W, rcP = rcQ/64, and rcQ is the number of data pairs in the  th two-dimensional grid. Considering mwNCC as an element, a nonlinear correlation matrix R is formed as:For a group of video sequences containing K-frame video sequence is expressed as:zλ denotes the zth eigenvalue of nonlinear correlation matrix R, z ∈ . Please note that the larger the NCIE, the higher the content continuity of the video sequence.Now, the overall correlation degree NCIE of RF denotes the number of redundant frames. The larger the DFR, the more frames are removed from the video sequence.The DFR is now mathematically expressed as:τ universal at different vehicle speeds in various scenes, a different τ is set to calculate the corresponding DFR and NCIE, respectively, for vehicle fast speed video (Fast video1) and vehicle slow speed video (Slow video1) containing suburban roads, as well as vehicle fast speed video (Fast video2) and vehicle slow speed video (Slow video2) comprising urban main roads; the results are shown in In order to make the threshold τ, the DFR also increases for all video sequences. However, please note that for the same threshold, the DFR corresponding to slow video is higher as compared to that of fast video.It is evident from τ ∈ , which is close to that at τ = 0, i.e., the case that no frame is removed; the NCIE drops sharply in τ ∈ ; the NCIE oscillates when τ > 2.5 but is generally much lower than that in τ ∈ . It is notable that there is an inflection point in τ ∈ , which leads to the mutation of NCIE, thus weakening the overall correlation of video sequence and causing video discontinuity.It can be seen from τ is further determined in the interval  so that it has a certain margin from the inflection point of NCIE to ensure the continuity of video and a high frame removal rate. τ ∈ .Therefore, the threshold τ = 2.1 and the video starts to become discontinuous. The NCIE is relatively stable in τ ∈ . Therefore, the optimal cosine angle threshold τop is selected to be 1.8, i.e., located in the middle of the stationary region with a certain margin from the mutation point. This ensures the minimum of frames to be fused under the premise of a continuous video sequence.It is evident from K frames, starting from the first frame, the first frame is regarded as the reference frame R and is retained. The second and third frames are regarded as the current frame C and the next frame F, respectively. The inter-frame difference RCθ between R and C and the RFθ between R and F are computed and compared with the optimal threshold τop to determine if the C is retained or redundant.For a group of video sequences containing RCθ and RFθ are both smaller than τop, C is regarded as a redundant frame and discarded. In this case, R stays the same in the next cycle. On the other hand, if RCθ is less than τop and RFθ is greater than τop, C is regarded as the retained frame, and serves as the reference frame R during the next iteration. At the same time, C and F both move backwards by one frame, until C is the last frame in the sequence. Here, C is selected as the retained frame and the frame selection process is terminated.If R, ZC, and ZF represent the sequence number of R, C, and F in the original video, respectively, count denotes the retained frame counter, and the sequence number of the retained frame is stored in array S. The frame selection strategy is expressed as follows:(1)S[0]←RZ←1;CZ←2;FZ←3;count←1;Initialization:(2)CZ < Kwhile {RC < τθop)   if (   {RFθ > τop)      if S[count + 1]←K.Keep the last frame and end the frame selection.Zτ.The above frame selection process ensures that the inter-frame difference between the retained frames is the highest, but not greater than the cosine angle threshold For the selected retained frame sequence, the improved IHS-curvelet algorithm  is used The anisotropy of the support interval of Curvelet transform is utilized to achieve an efficient expression of two-dimensional information. The automatic adjustment of low-frequency coefficient weight is adopted to avoid the halation information from contributing to the reconstruction.f  is the input of the Cartesian coordinate system, D represents discretization, l represents direction, k represents position, and j represents the scale of Curvelet decomposition.The brightness component I and the infrared image are decomposed by two-dimensional discrete Curvelet transform ,23. Theiα are mathematically expressed as:k1, k2) is the low-frequency coefficient of visible image, m is the critical value at the junction of halation and non-halation in the low-frequency coefficient matrix, n is the weight of the infrared low-frequency coefficient at the critical value m, l is the critical rate of change, reflecting the intensity of change in α at the junction of halation and non-halation. After multiple optimization and comparison, when m is 3, n is 0.75, and l is 2, the image fusion results reach the optimal level.The infrared low-frequency coefficient weights k1, k2) of the fused image is expressed as:k1, k2) is the infrared low-frequency coefficients.The low-frequency coefficient k1, k2) of the fused image adopts the modulus maximization for containing more detailed information:k1, k2) and k1, k2) are the high-frequency coefficients of the infrared image and the brightness component I, respectively.The high-frequency coefficient k1, k2) and k1, k2) are reconstructed by discrete curvelet transform in the frequency domain to obtain the new brightness component I′. The discrete Curvelet transform in the frequency domain can be expressed as follows:ω1, ω2] represents input in the frequency domain, and jl,k, is the Curvelet function in the frequency domain.The The IHS inverse transform is performed with the new brightness component I′, the original H and S. The resulting anti-halation fused image has very small halation and possesses rich details, such as edge contours and colors.In order to ensure that the frame number, frame rate, and playing duration of the fused video are consistent with that of the original video, this work considers the motion vector estimated by block matching search (MVEBMS)  as the bg which has the minimum matching error with the current block f in the reference frame range as the matching block, the relative displacement between g and f denotes the estimated motion vector MV. The block matching search is shown in SAD is expressed as:N × M = 16 × 16 pixels and search window have size ±8 pixels. i and j are the horizontal and vertical coordinates of pixels, respectively. xv and yv are the horizontal and vertical components of MV, respectively.Searching the block T of interpolated frames between them is:ZFR and ZBR are the sequence numbers of FR and BR, respectively. Considering the content difference FBθ between FR and BR as the reference, the adaptive weight iλ of the ith interpolation frame Si is expressed as:FSiθ is the content difference between FR and iS, 0 ≤ i ≤ T.Let us assume that the front frame of two adjacent retained frames is FR and the back frame is BR. Then, the total number MVSi of the interpolated frame iS can be expressed as follows:The estimated motion vector ith frame F to be interpolated is determined:x and y are the horizontal and vertical coordinates of the frame to be interpolated, respectively. xv and yv are the horizontal and vertical components of MViS, respectively. n and m are the relative coordinates of horizontal and vertical positions in the window function, respectively. j is the number of overlapping blocks ranging in . W  is the weight of pixel  in the window, expressed as:According to Equation (16), the pixel information of the interpolated frame is constructed. In order to satisfy the rate conversion and the visual effect of frame interpolation, the overlapping block motion compensation (OBMC) ,25 is seIn order to verify the effectiveness and universality of the anti-halation method based on infrared and visible video fusion, we collected infrared and visible videos on two typical roads covering common traffic at night, namely suburban roads and urban main roads. The videos include fast-speed vehicle videos and slow-speed vehicle videos in each scene.In the suburban road scene, there are almost no other light sources except vehicle beams, and the overall illumination is very low. In the urban main road scenario, the scattered light from streetlamps and surrounding buildings is weak, and the vehicles are using low beam lights. The halation areas of the videos increase in size and then shrink as the vehicles come closer. We have obtained more than 6200 original infrared and visible images from the videos collected.The experiments were performed using an Intel(R) Core (TM) i7-7700HQ CPU@2.80GHz , NVIDIA GeForce GTX1050 , and Windows8 64-bit operating system . The processing software sets include MATLAB2018a , Visual Studio 2017 , and OpenCV3.4.1 library .The proposed FSIFD and the retained frames extraction based on clustering (RFEC)  are compConsidering the Fast video1 and Slow video1 in suburban road scene as an example, the objective indicators of videos before and after frame selection are shown in It is evident from In slow video1, the proposed FSIFD discards 293 frames at the cost of a 0.2% reduction in NCIE, and the DFR is as high as 79%. The frames to be fused only account for 21% of the original video. The RFEC discards 307 frames at the cost of NCIE by 0.3%, and the DFR is as high as 83%.In fast video1, the NCIE only reduces by 0.3% when 144 frames are discarded in the proposed method. In addition, the DFR is 38%, and the frames to be fused is 62% of the original video. In the RFEC, 287 frames are discarded, and the DFR is as high as 77%, but NCIE is reduced by 2.9%. This is because the RFEC does not consider the motion between adjacent video frames, resulting in discontinuity and flickering of the selected frames.The above analysis shows that the proposed FSIFD ensures a high DFR on the premise of continuous video content at different vehicle speeds.It is also evident from The original visible and infrared images of a small halation area scene on an urban main road and a large halation area scene on suburban road are fused by the improved ©-Curvelet algorithm. The fusion results are shown in As presented in As presented in In summary, the halation of visible images seriously affects driving safety. The infrared image has poor visual effect when used to assist driving at night. The fused image is more suitable for human visual observation in the night halation scene.The proposed TMBAMC and the motion vector estimated by LSTMTA , OFFRN  and MVEBThe retained frame sequence of a slow video in a suburban road scene is considered as an example. The result of frame interpolation is shown in As presented in Among the 77 groups of interpolation results by LSTMTA, 57 groups are different from the original video, and the error rate is as high as 74%. In 32 groups, the interpolated frames are more numerous than in the original sequence, and the content is almost no different, resulting in a video stalling phenomenon. In another 25 groups, the interpolated frames are less numerous than in the original frames, which leads to a shorter playing duration and a video flicker phenomenon.OFFRN and MVEBMS have 46 and 59 groups of interpolation results that are inconsistent with the original video, respectively, among which 12 and 38 groups are more numerous than the original video, 34 and 21 groups are less numerous, and the error rate reaches 60% and 77%.This is because MVEBMS does not consider the influence of content differences between the adjacent reserved frames, and always inserts the fixed number of frames with the same content difference between two frames. OFFRN is based on the assumption that the object has a small displacement, and it has an ability to sense the motion changes between adjacent frames in slow video, so the number of inserted frames is more accurate than LSTMTA and MVEBMS.The proposed TMBAMC reconstructed frames according to time stamp, number of frames and playing duration of the interpolated video are consistent with that of the original video. Moreover, the interpolated frames have the same visual effect as the original sequence.When the number of frames inserted by four algorithms are consistent with that of the original video, the quality of the interpolated frame and the video continuity are further evaluated for the slow video in urban main road scene and the fast video in suburban scene.As presented in The position of the car in the first frame interpolated by the OFFRN is slightly ahead of the original frame 35. In comparison with frame 36, the position of the car and the amplitude of the pedestrian’s arm swing are both ahead in the second interpolated frame.The height of the trees in the first frame interpolated by the MVEBMS is lower than that of original frame 35, while the number of building floors is greater. In addition, the position of the car and the amplitude of the pedestrian’s arm swing lag behind frame 35. In comparison with frame 36, the position of the car and the amplitude of the pedestrian’s arm swing are both ahead in the second interpolated frame.This shows that the content of the interpolated frame is different from that of the original frame in LSTMTA, OFFRN and MVEBMS, and the effect is poor. The interpolated frames constructed by the proposed TMBAMC are almost similar to the original video in subjective vision, and the interpolated frames have higher quality.As presented in In conclusion, at different vehicle speeds in various scenes, the quality of interpolated frames constructed by the proposed TMBAMC is higher as compared to LSTMTA, OFFRN and MVEBMS in terms of subjective evaluation.α between the interpolated and the corresponding original frame. Ideally, when α = 0, the two frames are completely synchronized. In practice, it is considered that when α < 1, the content is synchronized; when α ≥ 1, the content is not synchronized; and when α > τop, it is discontinuous.In order to evaluate the quality of the interpolated frame and the continuity of the interpolated video objectively, the content synchronization is reflected by the vector angle The difference between the interpolated and the original frame of the slow video in urban main road scene is shown in α greater than 1 in the first 50 frames processed by LSTMTA, and the asynchronization rate between the interpolated frame and the corresponding original frame is as high as 44%. Especially, α reaches 1.82, 1.88, 1.87 and 1.89 in frame 21, 36, 49 and 50 respectively, and is greater than τop set by frame selection, which indicates that the video content is interrupted after frame insertion. The reason is that the quality of the subsequent interpolated frames is affected by the constructed interpolated frames in LSTMTA, which can cause error accumulation.It is evident from α in OFFRN is less than that in LSTMTA, and all values of α are less than τop, indicating that the content of interpolated frames is continuous. There are 7 frames in which α is greater than 1, and the asynchronization rate is 14%, which is 30% lower than that in LSTMTA. It indicates that OFFRN is superior to LSTMTA in the quality of frame interpolation for the slow video, meeting the assumption of small motion.The overall fluctuation of α reaches 1.88 and 1.81 in frame 14 and 35, respectively, and the video content is interrupted. α fluctuates greatly from frame 13 to 16, as well as from frame 35 to 36, is greater than 1, and the asynchronization rate is 12%, which is reduced by 32% and 2% compared with LSTMTA and OFFRN, respectively. The reason is that MVEBMS considers the motion between two adjacent reserved frames, so the effect of frame interpolation is more reliable.In MVEBMS, α fluctuates slightly in the whole sequence and is always less than 1 for the proposed TMBAMC, indicating that the content is synchronous and continuous, and the effect is more stable. The mean of α is 0.96, 0.68 and 0.86 in LSTMTA, OFFRN, MVEBMS respectively, while it is 0.54 in the proposed TMBAMC, indicating that the frames reconstructed by the proposed algorithm have higher quality. The reason is that the proposed TMBAMC estimates the motion vector based on the time mark and the content difference between the original frames, and have a higher expression for original video content, so the constructed frame is more similar to the original frame.However, The difference between the interpolated frame and the original frame of the fast video in the suburban road scene is shown in α is always less than 1, and shows small fluctuation in the whole sequence for the proposed TMBAMC, indicating that the video content is synchronous and continuous after frame interpolation.It is evident from α fluctuates greatly for LSTMTA, OFFRN and MVEBMS. The frames with α greater than 1 account for 46%, 54% and 30% of the total sequence, respectively, and the frames with α greater than τop account for 4%, 10% and 10%. Compared with the slow video in the urban main road scene, the discontinuous of video content and the stalling phenomenon are more serious. As a result, the effect of frame interpolation has further deteriorated.However, α in the LSTMTA, OFFRN and MVEBMS are 1.11, 1.18 and 0.97, respectively, while it is 0.57 in the proposed TMBAMC. Compared with the slow video in urban main road scene, the mean of α increases by 15.63%, 73.53%, 12.79% and 5.56%, respectively. Among them, due to the limitation of OFFRN based on ideal assumption, the frame insertion effect for the fast video is significantly different from that for the slow video, and its universality is low.The means of α is significantly smaller in the proposed TMBAMC, indicating that the algorithm has better quality of frame interpolation and stronger adaptability.From the above analysis, it can be obtained that the faster the vehicle speed, the lower the quality of the interpolated frames. Under the same conditions, the change in the mean of n) and space complexity S(n) of fusion video are shown in Considering the fast and slow videos in the suburban road and urban main road scenes as examples, the proposed method and the frame-by-frame fusion method are used to experiment the video fusion efficiency. The frame rate (FPS), time complexity T(n), T(n) of the proposed method is reduced by one magnitude compared with frame-by-frame fusion, indicating the proposed method effectively reduce the computational complexity.It is evident from As the driving speed is the same, there is little difference in the frame rate of fused video for various scenes, indicating that the fusion efficiency is less affected by the scenes. In the same scene, the fused video with faster speed has a lower frame rate. This is because the DFR of fast video is lower during frame selection, and there are more frames to be fused.The anti-halation method of video fusion proposed in this work effectively solves the lag caused by the frame-by-frame fusion of infrared and visible images. The designed frame selection fusion strategy discards the redundant frames to the greatest extent on the premise of continuous video content, reduces the number of frames to be fused, and improves the processing efficiency of video fusion. The proposed TMBAMC ensures that the video content is continuous and synchronized after frame insertion, and the duration is equal to that of the original video, which solves the phenomena of video stalling and flickering in the MVEBMS. The anti-halation method based on infrared and visible video fusion proposed in this work is applied to a night halation scene, which has a good halation elimination effect and helps to improve the efficiency of video fusion.


Malnutrition in older adults impacts health status, increased mortality, and morbidity. Malnutrition may increase the development of geriatric syndromes and contribute to a higher prevalence of falls and osteoporotic fractures that lead to loss of independence and an increased rate of institutionalization. The role of malnutrition in the pathogenesis of other geriatric syndromes seems to be well established. However, the data concerning nutritional interventions are confounding. Moreover, long-term undernutrition seems to be one of the factors that strongly influences the efficacy of interventions. This review outlines the current literature on this topic, and aims to guide physicians to make proper decisions to prevent the vicious cycle of falls, fractures, and their negative outcomes in patients with malnutrition. An older adult is typically described as an individual aged 65 years or older. Among this group, persons aged 75 years or older are usually differentiated, as they more often represent a specific profile of “geriatric patient” characterized by a higher degree of frailty, functional decline, and multiple active diseases . The gerThe aim of this narrative review was to estimate the problem of malnutrition and the risk of malnutrition in older adults and assess the influence of malnutrition on the risk of falls and fractures in clinical practice.Three specific features characterize geriatric syndromes: the prevalence of malnutrition is higher in this age group; the reasons for malnutrition are multifactorial; and health outcomes are more serious than in younger individuals. Malnutrition is one of the geriatric syndromes that greatly reduces regenerative and adaptive capacity, and remains an an important modulator of successful aging.The European Society for Parenteral and Enteral Nutrition (ESPEN) guidelines on clinical nutrition in geriatrics recommends using a combination of two features to diagnose malnutrition in oldest-old patients. The first is the phenotype criterion ) and the second one is the etiologic criterion . Experts also recommend diagnosing older patients who are at risk of malnutrition. The global consensus approach recommends diagnosing patients with higher malnutrition risk when oral intake is significantly reduced (≥50% of requirements for more than three days), or when risk factors that could reduce dietary intake or increase requirements  are present . The preUndernutrition in older people can be estimated using different tools, such as scales, anthropometric measurements, body composition parameters, and laboratory blood tests, as well as formulas such as the geriatric nutritional risk index (GNRI), which connects some laboratory tests and anthropometric features. Undoubtedly, in clinical research and in clinical practice, the first step should be the assessment of malnutrition risk, followed by the diagnosis of malnutrition. However, the two groups of geriatric patients should be given a regular, detailed estimation of malnutrition risk and diagnosis of malnutrition along with an assessment of other geriatric syndromes. Such other syndromes, such as functional decline, are crucial risk factors for malnutrition. Moreover, in the geriatric population, it is challenging to distinguish one factor that contributes to malnutrition  and multifactorial mechanisms that are most common .Scales used for the screening and evaluation of malnutrition in oldest-old patients in clinical practice include the Mini Nutritional Assessment Short Form and the Mini Nutritional Assessment Full Form , which are the most common scales in geriatric comprehensive assessment. Other available scales include the MST—Malnutrition Screening Tool, GNS—Geriatric Nutritional Status, and CONUT—Controlling Nutritional Status. However, the current ESPEN recommendations highlight that, beyond choosing the adjusted scale, the implementation of interventions based on effective tools plays a crucial role. Experts encourage clinicians to employ not only screening or evaluation, but also to apply interventions to improve clinical outcomes.Anthropometric measurements greatly contribute to the malnutrition assessment. Body mass index (BMI) is defined as the ratio of body mass (kg)/height (cm)2. The relationship between BMI and clinical outcomes has been recently discussed. It is probable that in older individuals, BMI alone is not enough to define malnutrition, and this phenomenon is more obvious with age. Some authors suggested the waist/hip ratio (WHR) was better, as it was more specific for abdominal obesity, whereas body mass index more reflects the lean body mass (LBM). However, WHR or waist (recommended now) seems not to be suitable for older adults, as the fluctuations strongly depend on body composition changes coexisting with musculoskeletal changes. The cutoff for BMI for adults is between 18.5 kg/m2 and 24.9 kg/m2, but for older adults, it should be greater—between 24 kg/m2 and 27 kg/m2 [Body composition (BC) estimation methods include measurements such as lean body mass (LBM), fat mass (FM), body fat, and percentage body fat (BF%). Body composition can be measured using computer tomography (CT), bioimpedance (BI), or dual-energy X-ray absorptiometry (DEXA)—a recommended method to assess BC. Assessing body composition in oldest-old patients has greater importance, especially when considering clinical outcomes, particularly the increased risk of sarcopenia and disability. Reduced muscle mass was one of the top five ranked criteria for diagnosing malnutrition recommended by GLIM. The other criteria were nonvolitional weight loss, low BMI, reduced food intake or assimilation, and disease burden or inflammation. In the published data, body composition remains more adequate for malnutrition phenotype assessment in older adults. Loss of skeletal muscle mass (SMM) is a physiological condition manifested by aging, a diminished whole-body metabolic rate, and a reduction in oxidative capacities. This contributes to lower energy expenditure from basal metabolism, which may lead to weight gain that is mostly fat mass. In succession, the accumulation of intramyocellular lipids promotes lipotoxicity and amplifies age-associated inflammation, mitochondrial dysfunction, and other metabolic changes such as insulin resistance (IR). Hormonal dysregulation, proinflammatory cytokines, myokines (including insulin-like growth factor 1 (IGF-1), myostatin, and irisin), and adipokines (including adiponectin and resistin) constitute crucial factors in muscle homeostasis and the pathogenesis of sarcopenic obesity. Furthermore, it is important to emphasize that a vicious cycle may occur in which a negative interaction is maintained not only between muscle and adipose tissue, but also with bone.Moreover, aging-related changes in body composition can affect the validity of the methods mentioned above. Regardless of the body mass index approach, the predicted BF% increases with age in both males and females. The appropriate contribution of FM and LBM to body weight is an essential risk indicator of major issues concerning geriatric care . The ranLaboratory tests can be useful in assessing qualitative and quantitative malnutrition. The most useful parameters available in everyday practice include serum albumin, prealbumin, total protein, transferrin, lymphocyte count, and level of total cholesterol, as well as vitamin D, acidic folium, and vitamin B12 serum concentration. The range of laboratory tests to assess micronutrient abnormalities and laboratory indicators of malnutrition for older adults have the same cutoffs as for adults. Among others, the evaluation of chosen laboratory tests, in addition to the chosen geriatric scale, has become an effective tool to predict postoperative functional status [l status . HoweverUndernutrition and/or malnutrition in the geriatric population is a challenging health concern related not only to increased morbidity and mortality, but also physical decline, which has wide range of implications for mobility, instrumental activities of daily living, and general quality of life. Malnutrition is common and may contribute to the development of geriatric syndromes. In older adults, it can be seen as either involuntary weight loss or, in some cases, a low BMI; however, hidden deficiencies, especially micronutrient deficiencies, are more difficult to assess, and they are commonly overlooked in clinical practice. In developed countries, the most common source of malnutrition is disease, while acute chronic disorders have a high potential to result in or intensify undernutrition. Thus, as aging processes are risk factors for developing malnutrition and strongly contribute to disease development; older adults are the highest-risk population for nutritional risk and/or malnutrition. Undoubtedly, the etiology of malnutrition is complex and multifactorial, but the exacerbation of this process is facilitated by aging processes. Our comprehensive narrative review summarized the current evidence on the determinants and assessment of malnutrition in old adults while considering the clinical perspective and the contribution of malnutrition to falls and fractures. It is challenging to understand, identify, and treat malnutrition when falls and osteoporotic fractures occur. In some cases, the treatment target may include supplementation of macro- and/or micronutrients when diet alone is not sufficient to meet age-specific requirements . The gerA wide range of consequences of malnutrition have a strong impact on the musculoskeletal system. The key role of undernutrition in the progress of sarcopenia is well established. Nevertheless, the correlation of malnutrition mechanisms influencing falls, fractures, and their outcomes is still an issue of debate, especially considering that observational and interventional studies have become more complex. The factors that play a crucial role in falls and fractures are long-term undernutrition, number of comorbidities, advanced age, and method of malnutrition assessment. p < 0.0001 and p = 0.26, respectively). The risk of harmful falls among malnourished patients were 8 times higher compared to the control group . The authors suggested that malnutrition assessment should be included in fall risk assessment to identify the high-risk group [The role of falls in health status is well established in the geriatric population. as they strongly affect losses in mobility and functional decline. Simultaneously, falls remain a key factor in osteoporotic fractures. The ratio of fractures is 7–9 times higher in older women with falls coexisting with osteoporosis or osteopenia, in comparison to women who only experienced falls or osteoporosis/osteopenia . For thisk group . Other rsk group . Despitesk group .Loss of bone mass constitutes one of the critical characteristics of osteoporosis. Simultaneously, it was reported that aging increases fat mass and decreases muscle mass in both men and women . A highe2, which constitutes the borderlines values for BMI for oldest-old patients. This association also was observed when adjusted with BMD, but the correlation was weaker [Johansson and colleagues conducted a meta-analysis that focused on the relationship between fracture risk and BMI in women. They found that there was a significant association between body mass index and FR for all osteoporotic fractures, as well as for hip fractures. However, the authors stressed that this association was observed in the range of BMI between the lowest values in the study population up to 25 kg/ms weaker . Neverths weaker .As the population ages, there is an increase in the number of patients who require treatment for age-related conditions, such as surgical treatment after femoral neck or trochanteric fractures. Such operations are indispensable for remaining physically active. Malnutrition is a problem with multifaceted consequences, and among them, a higher number of hospitalization negative outcomes in comparison to patients without malnutrition . The autSome authors aimed to identify effective interventions for malnutrition. It remains probable that skipping the first step—screening for malnutrition—is one of the key problems in undernutrition. Therefore, finding cases that assess the severity and planning intervention based on nutrition and exercises is indispensable, not only for hip fracture patients, but in all oldest-old patients with malnutrition to avoid lean mass loss. For patients admitted to the hospital after a hip fracture, screening for undernutrition is likely to be the first step in facilitating a fast nutritional intervention. Despite the wide range of observational studies confirming a strong relationship between malnutrition falls and fractures, the data concerning nutritional support are inconsistent. Some older double-blind, placebo-controlled randomized trials did not find any profit or found little benefit, but nutritional interventions in these studies was based on relatively low protein and calcium intake, and low compliance also was considered ,34. ThesExperts recommend a vitamin D supplementation dose of 800 to 2000 international units (IU) per day for adults who want to ensure a sufficient vitamin D status. These doses are also recommended for the treatment of vitamin D deficiency, but higher vitamin D doses  may be used for the first 4 to 12 weeks of treatment if a rapid correction of vitamin D deficiency is clinically indicated before continuing with a maintenance dose of 800 to 2000 IU per day. Treatment success may be evaluated after 6 to 12 weeks in certain risk groups  via measurement of serum 25(OH)D, with an aim to target concentrations of 30 to 50 ng/mL (75 to 125 nmol/L) ,37.A systematic review that focused on fall prevention in oldest-old patients based on recommendations published in the last three decades identified 15 high-quality practice guidelines for fall prevention and intervention . The recEnergy intake in aggressive nutrition therapy (ANT) is defined as total energy expenditure (TEE) plus the amount of energy accumulated (EA). The application and effect of ANT differs depending on the etiology and characteristics of malnutrition, and is performed in malnourished and/or sarcopenic individuals to increase body weight and muscle mass. Determinants such as advanced age, precachexia, and mild and moderate dementia are indications for aggressive nutrition therapy. Nevertheless, ANT is contraindicated when acute disease or injury with severe inflammation, severe dementia, and reduced activity occurs, and usually should be combined with aggressive exercise (including resistance training), which may constitute a real obstacle in clinical practice . MoreoveConcomitantly, ESPEN guidelines on clinical nutrition state that in older patients, malnutrition screening is necessary, and the recommended value for energy intake is 30 kcal per kg per day with at least 1 g of protein per kg body weight/day. Moreover, nutritional interventions should constitute a part of a multimodal and multidisciplinary intervention along with maintaining or increasing body weight to improve functional status and clinical outcomes. To achieve this aim, experts recommend, in older adults with malnutrition or who are at risk of malnutrition, fortified food and/or oral nutritional supplements that will provide 400 kcal/day or more, including at least 30 g of protein/day. The authors of these guidelines strongly recommended postoperative nutritional supplements in patients with hip fractures to reduce the risk of complications. A summary of the recommendations is presented in Nevertheless, some limitations of the accessible publications should be considered. The main limitation was the age of studied groups: usually they represented the younger geriatric population, which is not representative of older adults, and especially oldest-old patients. Moreover, other coexisting geriatric syndromes were not screened, and there was a wide variation in the methodologies of the programs used .Early detection of undernutrition, risk of falling, and fracture risk facilitate performing planned interventions, and is crucial in the geriatric population to prevent negative outcomes, functional decline, and institutionalization. Moreover, malnutrition contributes strongly to more harmful falls. Furthermore, patients with cognitive decline constitute the high-risk group for malnutrition and its complications. Hence, the systematic and individualized estimation of these factors can guide decisions regarding adjusted interventions. However, some studies showed that older adults estimated to be at risk of falling and/or malnutrition did not receive adequate interventions, and there are confounding data between assessed risk factors and planned and performed interventions . AccordiUndoubtedly, further studies are needed, especially focused on screening for the risk of malnutrition and malnutrition diagnosis in the older old population with musculoskeletal diseases.


ALDH2) is a carcinogenic acetaldehyde-degrading enzyme, and its low activity is a genetic constitution peculiar to East Asians. People with low alcohol dehydrogenase 1B activity (ADH1B*1/*1 genotype) have a high risk of developing head and neck cancer and alcoholism. The study aims to evaluate the effectiveness of brief interventions for excessive drinking among college students and adults in their 20s, including information on five constitutions that combine the ALDH2 and ADH1B genotypes.The alcohol-metabolizing enzyme aldehyde dehydrogenase 2 (ALDH2 and ADH1B enzyme activities); the result report will arrive approximately 1 month later. We will conduct a 30-min face-to-face or online intervention. The control group will be merely given the conventional materials, and genetic testing will be performed voluntarily after 6 months (end of study). The intervention group will undergo questionnaire surveys 1 month after the intervention and 3 and 6 months after baseline. Questionnaire surveys will be conducted 1, 3, and 6 months after baseline for the control group. The average amount of drinking before and after the intervention, attribute/baseline data between the two groups, and time-series data were compared using various analysis tools. For interventions, we engaged in dialog based on intervention materials that added genotyping content to the existing materials, result reports, baseline data, and drinking calendar records. Participants’ ingenuity is respected to support their drinking behavior and goal setting.Participants comprised university students and staff aged 20–30 years who had consumed ≥40 g  or ≥20 g  of pure alcohol; they were classified into intervention and control groups using a simple randomization method. Participants anonymously filled out questionnaires linked to identification numbers and recorded the drinking days and amounts on the drinking calendar. The intervention group will then be tested for genotype testing using saliva (5 types of combinations of ALDH2 low activity rate is high. This information may be useful for health care and precautionary measures.Individual information on the genetic makeup of alcohol-metabolizing enzymes provided during the intervention is more personal and objective than general health information, especially in Japan, where the R000050379, UMIN000044148, Registered on June 1, 2021.Scientific Title: Examination of simple intervention using genetic polymorphism information for excessive drinking. We ask the answer of the average number of drinks on the drinking day of the past month.A similar example is shown in the drinking calendar, and the drinking contents  and amount on the day of drinking will be recorded for half a year. From the record of the calendar, the sum of the drinking amount  and the number of drinks is calculated using the following generally accepted formula: amount of liquor (ml) × degrees or %/100 × 0.8 (specific gravity) = pure alcohol content (g).As the secondary outcomes, screening test (AUDIT-C and AUDIT) scores, awareness of participant’s flushing reaction (simple flushing questionnaire method two items) , recogniFurthermore, we will investigate the five stages of change in drinking behavior: precontemplation, contemplation, preparation, action, and maintenance . Self-reJapan’s Ministry of Health, Labor, and Welfare has listed support technology in the “Standard Health Checkup and Health Guidance Program Definitive Edition” and calls for “indifference, interest, preparatory period, execution period, and maintenance period” as “the preparation stage for behavior change,” and “to support the stage to improve by changing the support method for each stage”  . The fivThe modified PHCS Japanese version , which hBaseline data and participant attributes, average drinking amount on drinking days, AUDIT-C and AUDIT scores, and response at each scale of secondary outcome measures at each measurement point will be analyzed by a descriptive statistic.t-test, paired t-test, Wilcoxon rank-sum, chi-square, Mann–Whitney U, and Kruskal–Wallis tests as appropriate. The selection of the analysis method depends on the parametric and nonparametric data obtained from the results of descriptive statistics and normality tests to determine. In addition, the change in the average amount of drinking  in each group is examined by calculating the effect size. Data will be analyzed using IBM SPSS Statistics for Windows version 28. If the intervention is not possible in the intervention group, or if no response will be obtained after 1 month in the control group, it will be excluded from the analysis.Furthermore, baseline data, attributes, and changes in the average daily drinking amount  before and after intervention are compared with the differences between the two groups. We will also compare the measurement points within the groups and compare the groups in terms of genotypes. For these analyses, we use the Regarding qualitative data , we will examine them by using an inductive analysis method with qualitative coding. The code extracted to ensure reliability will be analyzed by multiple researchers, including those with qualitative research experience.Briefly, the intervention group will read the instructions for the alcohol-metabolizing enzyme test, sign the consent form, undergo saliva collection using a sponge swab from the test kit, and receive a report of the results approximately after 1 month. The inspection contractor will mail the result report to the participants. If face-to-face explanation and inspection are difficult to conduct, the test kit will be mailed to the participants.In the control group, testing will be conducted on those who wish to be tested after 6 months from baseline. After receiving the test results, their requests will be confirmed, and the intervention will be performed.Participants’ alcohol constitution Mechanism of alcohol decomposition, function of metabolic enzymes, and constitution by combinationRisks of diseases associated with drinking by alcohol constitution (every five types) and points to remember (comments by the constitution)How to interact with and drink alcohol that suits participants’ alcohol constitution, and how to absorb alcohol ingenuity in nutrition and eating and drinking to help slow down the metabolismDrinking habit screening test (AUDIT-C)Appropriate drinking and liver rest days, proper drinking method, alcohol calorie, and metabolism, obesity, etc.Inappropriate drinking: drinking while pregnant/lactating, drunk driving, alcohol harassment, and habitual use/heavy drinkingRegarding the report of the results of the alcohol-metabolizing enzyme test, we have created a pamphlet that includes contents common to each of the five types of constitutions and the contents for each constitution. The main contents are as follows:The intervening researcher is a nurse who had experienced caring for patients with health problems resulting from excessive drinking and who trained in intervention by physicians providing treatment to reduce drinking at the university. Specifically, this nurse will conduct simulated interventions with physicians online and face-to-face, receive advice (approx. 2 h), and observe physician’s interventions in outpatients (approx. 20 people).Participants in both the intervention and control groups can lead a normal life, with no particular restrictions or changes in daily care, which will continue in both study groups.Alcohol action Mechanism of comfort and discomfort associated with drinking Alcohol constitution Drinking habit screening test  and feedback that continuing the current drinking habits may eventually affect the mind and bodyDrinking amount ranking based on the current drinking amount Generally appropriate amount of alcohol: two drinks for males  or less on average per day, one drink  or less for femalesThe number of people who are vulnerable to alcohol, who are older, and who have a red face when drinking alcohol is even lessGenerally inappropriate drinking , minor, illnesses, or medications affected by drinking)Drinking with a high risk for lifestyle-related diseases: four drinks for males on average per day  or above, two drinks for females  or aboveBinge drinking: within 2 h, five drinks for males  or more, four drinks for females  or moreInformation from recent studies: zero daily drinking may be desirableParticipants’ intentions regarding drinking behavior and drinking habits (continuing or reviewing the current drinking habits is better)Participants’ opinion about the advantages and disadvantages of drinking alcoholReview on how to drink, the possible goal, and the concrete methodExamples of self-monitoring: proposal for continuing the drinking diary and drinking calendar recordingInterventions will be conducted according to the results of individual test results, baseline questionnaire data, drinking calendar records, and health guidance materials, including conventional teaching materials . The maiDuring the intervention, the intervening researcher confirms the participants’ perceptions of drinking behavior and drinking amount, their intention to review their habits, and the content they intend to change, and respects the participants’ ingenuity in drinking methods and goal setting.The intervention time is approximately 30 min as a guide, but it can be extended to 1 h to answer questions, share information with each other, and communicate. At the end of the intervention, the participants are told that they can contact the intervening researcher via e-mail for any questions or consultations, and they can respond to via e-mail as well, as needed. If an interview is requested after the intervention, we will set up an interview at the end of the half-year survey and only follow them up by e-mail. Given that measures against COVID-19 are required during the planning and implementation of this study, the application Zoom was used as the intervention method in accordance with the participants’ wishes and place of residence . Thus, the interview is conducted online or face-to-face. After the registration of participants had started, the progress was reported and the status was confirmed at a joint researcher’s meeting once a month.In the control group, the conventional teaching materials   will be A recall of experiences such as the loss of a child or parent resulting from a fatal accident or illness caused by drinking alcohol, or a dangerous experienceCurrent drinking style of university students (results of the drinking attitude survey)Conversion of the amount of alcohol consumed on the day of drinking according to a conversion table and checking of the current amount of alcohol consumed by using the alcohol consumption ranking table Stage from tipsy to death resulting from acute alcohol intoxicationBinge drinkingChecking, defining, and coping with alcohol and harassmentSpecific examples of ideas for keeping tipsy at drinking partiesKnowledge of alcohol The main contents of the conventional materials are also included in the abovementioned test result report and health guidance materials for intervention. Although some descriptions are duplicated, the main contents are as follows:This research began in July 2021, with the approval of the Institutional Review Board of Medical Doctors, University of Tsukuba . If there are any changes in the research protocol, the principal researcher will apply to the Institutional Review Board of Medical Doctors, University of Tsukuba  for approval.The researcher strictly manages the correspondence table that collates the participants with the ID number, and the data are managed so that the participant’s name and the information on the genotype constitution heard are not directly linked. Data input and analysis are mainly performed by the first author using a computer with security measures, and data collection and analysis results are regularly checked with collaborators.This research was conducted in compliance with the “Ethical Guidelines for Human Genome/Gene Analysis Research” established by the Ministry of Health, Labor, and Welfare. Registration for the UMIN clinical trial was completed on June 1, 2021. The results of this research will be published in peer-reviewed academic journals and at internal and international academic conferences.Although this study is an intervention study, it is an educational intervention, and saliva testing is not considered invasive. Therefore, there is no data monitoring. To maintain compliance, we collaborate with researchers to attend monthly online meetings to share progress, data collection, and intermediate analysis results and discuss issues. Regarding the decision to suspend the recruitment of participants, the progress of the trial will be shared at the meeting, and if there is no problem in conducting the exam, the recruitment will be suspended when the number of subjects to be analyzed reaches the target. This research is subject to progress checks and audits every 6 months by the Institutional Review Board, and the person in charge of the audit is independent of this study.Participation in this study is considered of little risk. However, 6 months of cooperation surveys, answering questionnaires, and receiving interventions are a small burden. As a benefit of participation, knowing the genetic predisposition of alcohol-metabolizing enzymes can lead to better management of drinking behaviors and reduced risks associated with drinking. On the contrary, information about the health risks associated with drinking may lead to participant anxiety. We will provide contact information for follow-up consultation in the case of mental burden. In the unlikely occurrence of an adverse event, participants will be instructed to contact the researcher, who will notify the responsible researcher’s doctor and take appropriate action, and will report to the department of the Institutional Review Board of Medical Doctors, University of Tsukuba  as necessary.ALDH2 activity (a genetic component peculiar to East Asians) is high [ALDH2 polymorphism has been elucidated as follows [ALDH2 * 2 allelic carriers have a higher risk of carcinogenesis than non-ALDH2 * 2 but are less likely to increase blood pressure and have relatively lower serum liver transaminase  and uric acid levels.Information on the genetic function and constitution of alcohol-metabolizing enzymes provided individually by the study’s intervention may lead to a transition to a period of interest, such as a partial review of drinking methods, from indifference that participants perceive alcohol drinking as personal and objective information rather than general health information and are not aware of alcohol or hazardous drinking. At least in Japan, where the proportion of low  is high , 2, gene follows : ALDH2 *ALDH2 * 2 allelic carriers may not have an opportunity to monitor their drinking habits [Moreover, g habits . Therefoalcohol dehydrogenase 1B (ADH1B * 1 / * 1 genotype) have been reported to be at an increased risk of alcohol dependence [ALDH2 genotype information [ALDH2 and ADH1B gene polymorphisms into five combinations in an easily comprehensible manner. However, the effectiveness of health guidance using the information on ALDH2 and ADH1B gene polymorphisms needs to be verified by increasing the sample size. Regarding the strength of this study, we will investigate the intervention effects of new efforts using the method of randomized controlled trials.As previously mentioned, carriers of pendence , as wellpendence –8. Previormation , 19. In ALDH2 validated the effect 30 days after providing Internet-based ALDH2 genotype and health risk information [ALDH2 activity. However, their follow-up period was 30 days, which is shorter than 6 months (maintenance period) [As mentioned in the introduction, a randomized controlled trial using the genetic information of the alcohol-metabolizing enzyme ormation . In this period) , for beh period) , which iConversely, one limitation of this study is that when students agree to participate, they already know if they are in the group to be tested or not; hence, they cannot be blinded. Regarding the primary outcome measure, the average drinking amount on the drinking day is limited to the calculation of the amount of drink with an unknown alcohol concentration or amount. In addition, given that this research deals with genetic information, we need to evaluate the intervention effect by interviewing each participant. We also need to examine a system that can be useful for enrolling large groups, such as university students.This study will contribute to developing intervention methods, including the improvement of lifestyle habits using genetic polymorphic information for managing drinking problems.https://www.umin.ac.jp/ctr/index.htm, R000050379) on June 1, 2021. The present protocol version 2 was changed to correct misdescriptions on April 16, 2022. The first participant was registered on July 20, 2021. Recruitment will be completed on March 31 2023.The study was firstly registered in UMIN Clinical Trials Registry (


TIPARP-upregulated human trabecular meshwork (HTM) cells. Moreover, western blot analysis showed that collagen types I and IV, fibronectin, and α-SMA were increased in TIPARP-downregulated or TIPARP-inhibited HTM cells. In addition, cross-linked actin networks were produced, and vinculin was upregulated in these cells. Subconjunctival injection of the TIPARP inhibitor RBN-2397 increased the IOP in Sprague–Dawley rats. Therefore, we identified TIPARP as a regulator of IOP through modulation of extracellular matrix and cell cytoskeleton proteins in HTM cells. These results indicate that TIPARP is a potential therapeutic target for ocular hypertension and glaucoma.Elevated intraocular pressure (IOP) is the major risk factor for glaucoma. The molecular mechanism of elevated IOP is unclear, which impedes glaucoma therapy. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-inducible Poly-ADP-ribose Polymerase (TIPARP), a member of the PARP family, catalyses mono-ADP-ribosylation. Here we showed that TIPARP was widely expressed in the cornea, trabecular meshwork, iris, retina, optic nerve, sclera, and choroid of human eyes. The expression of TIPARP was significantly upregulated in the blood and trabecular meshwork of patients with primary open angle glaucoma compared with that of healthy controls. Transcriptome analysis revealed that the expression of genes related to extracellular matrix deposition and cell adhesion was decreased in  TIPARP, an inducible poly-ADP-ribose polymerase, is identified as a novel regulator of intraocular pressure through modulation of extracellular matrix and cell cytoskeleton proteins in trabecular meshwork cells. Primary open angle glaucoma (POAG) is the most common type of glaucoma and is a complex disease with genetic heterogeneity1. Elevated intraocular pressure (IOP) is the major and only modifiable risk factor for POAG2. IOP is determined by the balance between aqueous humor secretion and drainage mainly through the conventional outflow pathway consisting of the trabecular meshwork, juxtacanalicular tissue and Schlemm’s canal3. Abnormally high resistance to aqueous humor outflow induced by trabecular meshwork malfunction is an important aetiology of POAG6. However, the underlying molecular and cellular mechanisms of increased resistance to aqueous humor outflow still need to be explored.Glaucoma is a group of diseases characterized by progressive degeneration of the optic nerve and is the top reason for irreversible blindness worldwide7. As a member of the PARP family, TIPARP transfers mono-ADP-ribose from nicotinamide adenine dinucleotide (NAD+) to substrates and catalyses mono-ADP-ribosylation, which is an important and ubiquitous post-translational protein modification8. TIPARP has important functions in many physiological processes, including gene regulation, viral response, and cytoskeleton regulation9. The dysregulation of TIPARP may induce neuronal developmental disorders, abnormal antiviral responses, some types of cancer, and dioxin-induced steatohepatitis16. Evidence of the association between TIPARP and POAG has been reported in a few studies. A meta-analysis of more than 13,000 Europeans revealed that the SNP rs9822953 located in TIPARP is significantly associated with central corneal thickness, which is a risk factor for POAG17. In a glaucomatous lamina cribrosa cell model induced by hypoxia stress, the expression of TIPARP was significantly increased18. However, the role and regulation of TIPARP in trabecular meshwork cells and aqueous humor outflow are unknown.TIPARP (TCDD-inducible poly-ADP-ribose polymerase), also known as PARP7 or ARTD14, is a TCDD -induced poly-ADP-ribose polymerase (PARP)To obtain greater insights into the relationship between TIPARP and IOP regulation, we examined TIPARP expression in patients with POAG and explored the function and molecular mechanism of TIPARP in aqueous humor outflow regulation. This study suggested that TIPARP participated in IOP regulation by modulating extracellular matrix (ECM) deposition, cell adhesion and cross-linked actin network (CLAN) formation.19. After 5 days of treatment with dexamethasone, the myocilin expression of HTM cells was increased according to immunofluorescence  was significantly higher than that in the controls (n = 35)  compared with that in the control human donor eyes  and 50.21% ± 1.50% in HTM cells transduced with TIPARP-encoding lentivirus (LV-TIPARP HTM cells)  and LV-GFP HTM cells . The Gene Ontology biological process analysis of the differentially expressed genes revealed that biological adhesion, cell junction, ECM, and extracellular region were associated with TIPARP upregulation , laminins , elastin and fibrillin , fibulins , and microfibril-related genes . Genes participating in cell-cell and cell-ECM adhesion  were reduced in the LV-TIPARP HTM cells . However, the expression of different members of the matrix metalloproteinase family did not show the same trend. Western blot analysis confirmed the downregulation of collagen type IV, fibronectin and α-SMA in the LV-TIPARP HTM cells  Fig. . The TIPion Fig. . Specifills Fig. . In addills Fig. .Fig. 3Tr15. The cocrystal structure of RBN-2397 showed that it can bind to the NAD+-binding pocket15. RBN-2397 treatment did not alter the expression level of TIPARP in HTM cells , while the control cells contracted 18.33 ± 3.32 percent of their initial size after 24 h of treatment (n = 5)  in HTM cells. The mRNA and protein expression levels were decreased by treatment with TIPARP siRNA compared with the scrambled siRNA control , which indicated excessive ECM deposition and pathologic profibrotic responses under TIPARP deficiency. Moreover, following downregulation of TIPARP expression, the formation of CLAN structures increased, which could increase the stiffness of HTM cells . After the second injection on Day 7, the IOPs of the RBN-2397-treated group increased steadily higher than those of the vehicle control group from Day 8 to Day 14   was injected into the subconjunctival space of 01) Fig. . On Day 01) Fig. .Fig. 7EfTIPARP-upregulated HTM cells. We demonstrated that loss of TIPARP function caused ECM deposition, CLAN formation and focal adhesion. In addition, we found that the inhibition of TIPARP resulted in elevated IOP in rats. In conclusion, our results show that TIPARP participates in the pathogenesis of POAG.In the present study, we identified a key role for TIPARP in the regulation of trabecular meshwork function and IOP. We investigated the expression of TIPARP in the eye, and found that the TIPARP expression levels were increased in the trabecular meshwork and blood in the patients with POAG compared with the controls. These findings indicate the critical role of TIPARP in IOP regulation. Then, we showed that TIPARP participated in the regulation of ECM components and cell-ECM adhesion by utilizing 20. Some PARPs, including PARP1, catalyse poly ADP-ribosylation and have been well studied9. Other PARPs, including TIPARP, also known as mono(ADP-ribosyl) transferase (MART) enzymes, catalyse mono-ADP-ribosylation (MARylation), which is not well understood9. As a MART enzyme, TIPARP catalyses the mono-ADP-ribosylation of target proteins and exerts effects on the viral response, transcriptional regulation, stem cell pluripotency, neuronal function and cytoskeleton regulation14. In addition, TIPARP has been identified as a potential target of cancer therapy16. Although the importance of TIPARP has been gradually discovered in recent years, research on its function remains very limited.The PARP family is a group of enzymes catalysing cellular ADP-ribosylation, which is an important post-translational protein modification. ADP-ribosylation is involved in many biological processes, such as oxidative stress, DNA repair, cytoskeletal regulation, transcriptional regulation, and immune cell function7, but the distribution of TIPARP in the eye has yet to be described. We showed that TIPARP was expressed widely in eye tissues, including the cornea, trabecular meshwork, iris, sclera, choroid, retina, and optic nerve. In addition, we demonstrated that TIPARP expression was upregulated in patients with POAG and in eyes perfused ex vivo. The distribution of TIPARP in the ocular drainage structures and the abnormal expression of TIPARP in the patients with POAG suggested that TIPARP might play a regulatory role in maintaining trabecular meshwork function and normal IOP.Our study focused on the role of TIPARP in IOP regulation. TIPARP is expressed in many mouse tissues, including the heart, brain, lung, liver, spleen, and reproductive organs21. Together, these proteins are involved in IOP regulation. The function of PARPs and the role of ADP-ribosylation in aqueous humor outflow are still unclear. We found that a variety of genes associated with ECM and focal adhesion were downregulated in the TIPARP-overexpressing HTM cells by RNA-seq. Previous studies have demonstrated that the trabecular meshwork regulates aqueous humor outflow resistance by cells and their ECM interplay and ECM protein interactions6. Abnormal changes in the ECM have dramatic effects on trabecular meshwork function23. The Gene Ontology analysis of RNA-seq identified ECM, ECM-related genes, biological adhesion, and cell junction as key pathways impacted by TIPARP upregulation. The expression of many ECM component genes, including collagen , elastin and fibrillin (ELN and FBN1-2), laminins (LAMA2), and fibulins (FBLN1-2 and 5), was decreased. The ECM deposition regulatory genes, including LTBP1 and LOXL1, and the cell adhesion genes ITGA1 and VCL also showed decreased. In addition, the expression of most tissue inhibitor of metalloprotease family members was downregulated , which was consistent with the expression changes in ECM gene expression. Therefore, TIPARP might be involved in the maintenance of IOP by regulating ECM deposition and cell adhesion. To further confirm the function of TIPARP, we inhibited and downregulated TIPARP in HTM cells. Consistent with the RNA-seq results, increased ECM deposition, cell adhesion, and CLAN formation were observed in the TIPARP-inhibited or downregulated HTM cells. Excessive ECM deposition has been observed in the POAG outflow pathway, which results in tissue stiffness and increases the resistance to aqueous humor outflow24. The actin cytoskeleton is another key factor in the regulation of outflow resistance. The abnormal arrangement of the cytoskeleton and formation of CLAN affect trabecular meshwork contraction and then influence outflow resistance25. In addition, an abnormal increase in α-SMA indicates pathologic fibrotic responses that produce myofibroblast-like contractile cells and increase tissue stiffness28. Then, the in vivo experiment provided further validation of TIPARP regulating IOP. After subconjunctival injection of the TIPARP inhibitor RBN-2397 in rats, the IOP of the rats was elevated. Taken together, these in vitro and in vivo experiments confirmed that TIPARP was involved in IOP regulation by regulating the ECM, actin cytoskeleton and cell adhesion.We further clarified the role and mechanism of TIPARP in IOP regulation through in vitro and in vivo experiments. The main glaucoma-related events occurring in trabecular meshwork include oxidative stress, mitochondrial impairment, endothelial dysfunction, and proteome changes in the aqueous humor, which includes mitochondrial proteins, cell adhesion proteins, protein kinases, and neuronal proteinsTIPARP was reported to reduce cell growth, migration, and invasion in OVCAR4 cells12. The functions of TIPARP target proteins identified by mass spectrometry were enriched in cell-ECM adhesion and cytoskeleton organization12. Similarly, Tiparp–/– mice presented a decrease in neural progenitor cell proliferation and a reduction in neural stem cell migration because of the abnormal mono-ADP-ribosylation levels of cytoskeletal proteins13. Our results supported these studies about the roles of TIPARP in cell-ECM adhesion and the cell cytoskeleton and added additional roles of TIPARP in ECM deposition.Previous studies have also demonstrated that TIPARP and its mono-ADP-ribosylation are important targets for regulating the cell cytoskeleton and cell adhesion. In a study on ovarian cancer, knockdown of TIPARP might play a positive role in IOP regulation in POAG. Previous studies have shown that a series of mechanisms exist to maintain IOP homeostasis29. For example, the levels of some matrix metalloproteinases, important ECM remodeling regulatory factors, are increased in the aqueous humor of patients with POAG31. Similarly, the upregulation of TIPARP could partially reduce the aqueous humor outflow resistance and help to re-establish the IOP balance. In the future, the development of agonists or analogs of TIPARP may be a new method to reduce IOP.Interestingly, TIPARP expression was upregulated in blood samples and the trabecular meshwork of our patients with POAG. Furthermore, an ex vivo mouse eye perfusion model showed that elevated intraocular pressure could lead to the upregulation of TIPARP expression in outflow tissues of mouse eyes, which was consistent with the upregulation of TIPARP in patients with POAG. Combined with the results of a series of in vitro and in vivo experiments on TIPARP functions, we conclude that the increased expression level of One of the deficiencies of our study is the comparability of trabecular meshwork tissues between patients with POAG and controls. Trabeculectomy for the patients with POAG was performed by an experienced ophthalmologist to ensure that trabecular meshwork tissue was included. The trabecular meshwork tissues from the control donor eyes were isolated using a similar method to ensure that the tissues of the two groups had similar compositions. The cadaveric trabecular meshwork tissues were dissected within 12 h after death from our eye bank. Prolonged time could affect the freshness of cadaveric trabecular meshwork tissues, which might affect the expression of TIPARP. Therefore, TIPARP expression in the blood of the patients with POAG and the controls was tested to provide additional evidence.In conclusion, our study illustrated the key role of TIPARP in the regulation of trabecular meshwork function and IOP. The expression level of TIPARP was elevated in patients with POAG. Downregulation or inhibition of TIPARP could elevate IOP by increasing ECM deposition, producing CLAN and increasing cell adhesion. Thus, TIPARP is a potential target of IOP regulation in POAG.32. In brief, the inclusion criteria for patients with POAG were as follows: (1) IOP > 21 mmHg; (2) glaucomatous visual field defects and glaucomatous optic disc damage; (3) open anterior chamber angle; (4) absence of any secondary glaucoma; (5) absence of other ocular diseases that could severely affect visual acuity; (6) brain/orbit magnetic resonance imaging showing no compression lesions. Inclusion criteria for controls were as follows: (1) absence of a history of glaucoma or elevated IOP; (2) absence of very narrow angle; (3) a vertical cup-disc ratio of ≤ 0.5; (4) absence of ocular disorders that could severely affect the visual acuity or visual field; (5) absence of a family history of glaucoma. Trabecular meshwork tissues of the patients with POAG were collected from trabeculectomy surgeries, and control eyeball tissues were obtained from the Eye Bank of the Eye & ENT Hospital, Fudan University. Trabeculectomy for patients with POAG was performed by an experienced ophthalmologist to ensure that trabecular meshwork tissue was included. The trabecular meshwork from the control donor eyes was isolated using a similar method to ensure that the tissues of the two groups had a similar composition. Trabecular meshwork tissues from ten patients with POAG were pooled for protein extraction. Trabecular meshwork tissues from four control donor eyes were pooled for protein extraction. The expression of target proteins in outflow tissue was measured by Western blot. Another group of patients with POAG (n = 32) and controls (n = 35) were enrolled to test TIPARP expression in blood.All procedures of this study were approved by the Institutional Review Board of the Eye & ENT Hospital of Fudan University. Informed consent was obtained from all participants. All patients with POAG and controls recruited in our study met the inclusion/exclusion criteria described in our previous study2, according to the manufacturer’s instructions. For HTM cell characterization, dexamethasone (500 nM) was added to the HTM cell medium for 5 days. The control group was cultured without dexamethasone. The expression of myocilin was measured using both Western blot analyses and immunocytochemistry.One strain of primary HTM cells, purchased from ScienCell , was cultured in Trabecular Meshwork Cell Medium  at 37 °C under 5% COProteins were extracted from tissues or cells using RIPA lysis buffer (Beyotime). After centrifugation, the supernatant was mixed with 5X loading buffer (Beyotime) and boiled. The denatured proteins were separated by 4–12% or 4–20% SDS polyacrylamide gel (GenScript) and electrophoretically transferred to PVDF membranes (Millipore). The membranes were blocked with 5% nonfat dry milk for 1 h and probed with antibodies against TIPARP  and 0.1% Triton X-100 (Sigma-Aldrich) for 2 h at room temperature. The sections were incubated with primary anti-TIPARP antibody  overnight at 4 °C, washed three times in phosphate buffer saline, and then incubated in Alexa Fluor-555 or Alexa Fluor-488 goat anti-rabbit secondary antibody  for 2 h at room temperature. Nuclei were visualized by DAPI  counterstaining.HTM cells, which grew on 0.1% gelatin-coated glass coverslips in 24-well plates, were fixed with 4% paraformaldehyde for 15 min and then blocked and permeabilized with 5% bovine serum albumin and 0.1% Triton X-100 for 1 h at room temperature. Then, the coverslips were incubated with anti-TIPARP  and anti-vinculin  antibodies overnight at 4 °C and Alexa Fluor-488 or Alexa Fluor-555 goat anti-rabbit secondary antibody  for 2 h at room temperature. TRITC-phalloidin Atto 565 (Sigma-Aldrich 94072) was used for F-actin staining for 40 min at room temperature. DAPI staining was performed at room temperature for 5 min.34. At least three imaging fields were evaluated in each group. The ratio of the number of cells containing CLAN structures to the total number of cells was evaluated34.All primary antibodies, secondary antibodies and DAPI were diluted in phosphate buffer saline containing 1% bovine serum albumin and 0.1% Triton X-100. An inverted confocal microscope (Leica) was used for imaging. The minimum structure of CLAN was defined as three hubs connected by three actin spokesTIPARP mRNA expression was measured by a SYBR Green quantitative real-time PCR kit (TaKaRa) according to the manufacturer’s protocol and analyzed on a ViiA 7 Real-Time PCR System (Life Technologies) or CFX96 Real-Time PCR System (Bio-Rad Laboratories). The primer sequences used were as follows: human TIPARP primer F 5’GGCAGATTTGAATGCCATGA3’, primer R 5’TGGACAGCCTTCGTAGTTGGT3’; human β-actin primer F 5’TTGTTACAGGAAGTCCCTTGCC3’, primer R 5’ATGCTATCACCTCCCCTGTGTG3’. The relative expression level of mRNA was normalized to that of the reference gene β-actin. The results were calculated by the comparative CT method (2-ΔΔCT).Total RNA was extracted from whole blood using a RNeasy Mini Plus Kit (Qiagen), and total RNA from HTM cells was extracted using an EZ-press RNA purification kit (EZ-Bioscience) according to the manufacturer’s protocol. The RNA concentration was measured using a NanoDrop spectrophotometer. Reverse transcription was performed using a reverse transcription kit (TaKaRa). TIPARP. Lentivirus-GFP was used as a control. Lentivirus-GFP-TIPARP and lentivirus-GFP were produced by Hanheng Company . According to the manufacturer’s instructions, lentivirus-GFP-TIPARP and lentivirus-GFP were diluted in complete medium containing 8 µg/mL polybrene at a final multiplicity of infection (MOI) of 15. For knockdown of TIPARP expression, HTM cells were transfected with siRNA specific to human TIPARP (TIPARP siRNA) using Lipofectamine 2000 (Invitrogen/Thermo Fisher) according to the manufacturer’s instructions. The control cells were transfected with scramble control siRNA (scrambled siRNA). TIPARP siRNA and control siRNA were designed and produced by GenePharma Biological Company . Quantitative real-time PCR and western blot were performed to measure TIPARP expression after upregulation or downregulation.HTM cells were transduced with lentivirus-GFP-TIPARP to upregulate the expression of TIPARP, we performed RNA-seq in TIPARP-upregulated HTM cells. After transduction with lentivirus-GFP-TIPARP or lentivirus-GFP for 96 h, the HTM cells were collected for RNA isolation. TRIzol reagent was used for total RNA extraction according to the manufacturer’s protocol. RNA purification, quantification and integrity were evaluated using a NanoDrop 2000 spectrophotometer (Thermo Scientific) and an Agilent 2100 Bioanalyzer (Agilent Technologies). The libraries were constructed by a TruSeq Stranded mRNA LT Sample Prep Kit (Illumina) and sequenced on an Illumina Novaseq 6000 platform. After the raw reads obtained by sequencing were filtered, the clean reads were mapped to the human genome (GRCh38) using HISAT235. The FPKM36 of each gene was calculated by Cufflinks37, and the read counts of each gene were obtained by HTSeq-count38. The DESeq2 was used for differential expression analysis39. The threshold for significantly differential expression was q value < 0.05 and fold change > 2 or fold change < 0.5. Hierarchical cluster analysis of differentially expressed genes was used to show the expression pattern of genes in different samples and groups. Gene Ontology enrichment and KEGG pathway enrichment analyses of differentially expressed genes were performed using R40.To determine the possible effects of 6 cells/mL were mixed with collagen gel and added to a 24-well plate. After 1 h of incubation at 37 °C for polymerization, the culture medium was added to each well of a plate. After 48 h of incubation, the edge of the gel was carefully detached using a syringe needle. After 13 h, the gel area did not change, and natural contraction was observed. Then, 1 ml of the TIPARP small-molecule inhibitor RBN-2397 (1 μM) was added to the medium. The images were captured at 24 h. The gel area was calculated using ImageJ.HTM cell contraction was tested by a cell contraction assay kit  following the manufacturer’s instructions. In brief, HTM cells at a density of 2 ×10Sprague–Dawley rats and C57BL/6 J mice were bred and housed in standard cages with a 12 h-12 h light-dark cycle. Male rats aged approximately 8 weeks and weighing 180–200 g were used. Male mice aged 8–10 weeks were used.All animal experiments were conducted under the ARVO statement for the Use of Animals in Ophthalmic and Vision Research. The experimental protocols were approved by the Institutional Review Board of the Eye & ENT Hospital of Fudan University. 41. After perfusion, conventional outflow tissues of mice were dissected using an established method42. Outflow tissues from three eyes were pooled for protein extraction, and TIPARP expression was measured by Western blot.For determination of the effect of IOP elevation on TIPARP expression, mouse eyes were perfused with phosphate buffer saline at constant pressures of 6 and 18 mmHg for 4 hAll rats were deeply anaesthetized intraperitoneally with ketamine (80 mg/kg) and xylazine (5 mg/kg). Then, 25 μL of 10 µM RBN-2397 was slowly injected under the upper and lower conjunctiva of the eye respectively with a 33-gauge syringe . As a vehicle control, 0.1% dimethylsulfoxide in phosphate buffer saline was injected into the contralateral eye. The injection was repeated on Day 7. IOP measurements were performed before subconjunctival injection and every day after injection. IOP was measured using rebound tonometry  between 10:00 a.m. and 12:00 p.m. in live animals. Conventional outflow tissues of rats were dissected on Day 14. Outflow tissues from two eyes were pooled for protein extraction. The expression of target proteins in outflow tissue was measured by Western blot.t-test was used for analysis if the data were normally distributed. The Mann‒Whitney U test was used for analysis if data were not normally distributed. The statistically significant P value was set as 0.05.Statistical analyses were performed with SPSS 20 . All experiments were repeated at least three times. Numerical data were presented as the mean ± standard deviation. A Further information on research design is available in the Supplementary InformationDescription of Additional Supplementary FilesSupplementary Data 1Reporting Summary


Paired t-test was used to assess their change in knowledge of passive and forced drinking, and health and social harm of drinking after, the health talk. McNemar’s chi-squared test and adjusted multivariable logistic regression (AOR) were used to assess their change in intention to drink and intention to quit. Students were less likely to drink  and more likely to quit drinking  after the health talk. Increased knowledge of passive drinking was associated with less intention to drink , increased knowledge of health harm , and social harm of drinking . Similar associations were observed in forced drinking . We showed preliminary evidence that the health talk on passive and forced drinking reduced the intention to drink in adolescents.Passive and forced drinking harm was prevalent but less recognized in Chinese adolescents. We educated adolescents on such harm to reduce their intention to drink. Students ( It cont = 1087) . Alcohol12-month . Althoug12-month  in adoleThe adverse effects of alcohol are not limited to drinkers themselves but also people surrounding the drinkers. Passive drinking is defined as harm resulting from others’ drinking; serious harm includes sexual harassment, unwanted intercourse, and physical assaults by the drinking person . AdolescForced drinking is defined as drinking alcohol unwillingly, including drinking under force, playing drinking games, and drinking due to social pressure . DrinkinCochrane systematic reviews found that school-based prevention programs were cost-effective in delaying the onset of harmful drinking in adolescents even with small effect sizes, and saved substantial government expenses on alcohol-related morbidity ,19,20. Yn = 1244) were analyzed. Ethical approval was granted by the Institutional Review Board of the University of Hong Kong/Hospital Authority Hong Kong West Cluster (Ref. UW 19-400).In this quasi-experimental study, we used the Education Bureau (EDB) 2018 list of secondary schools as the sampling frame. The target sample size was 1230 students from 10 public schools. We recruited local secondary schools from all five regions of Hong Kong  using random sampling method and invited schools to join. Seven public schools with the same school ranking from each of the five regions participated. School participation was voluntary, with refusal mainly due to time and administrative issues. All Students in forms 1–6 (Grade 7–12), in the participating schools, were invited to the video-based health talk. Invitation letters were sent to parents via the schools to explain the health talk’s purpose and emphasize that participation was voluntary. Parents who refused to join were asked to have their children return blank questionnaires after the talk. Students’ participation was voluntary, even with parental consent. Before the health talk, students were briefed about the self-administered anonymous questionnaires, and teachers were reminded not to influence students to answer the questions. The two surveys (pre- and immediate post-questionnaire) took 5–10 min to complete, and 10 min break was given to students before completing the post-questionnaire. The completed questionnaires were inserted into opaque envelopes and sealed immediately after completion. Students who completed both questionnaires . Correct answers were provided and explained after each vote, and small gifts were given to students who voted for the correct answers. In the second session, students were introduced to passive drinking and forced drinking concepts, including the harm, situations, risk factors, and strategies to avoid those harm. We created two age-appropriate cartoon animations related to the knowledge of passive and forced drinking. The videos educated students on the harm of passive drinking (a story about a drunk driving victim) and strategies to avoid forced drinking (ways to avoid drinking invitations from peers). Each video lasted 2 min, and were shown in the health talk to enhance students’ impressions. In the final session, students were taught the strategies to reject peers’ or family members’ drinking invitations . Important points of the above information were summarized at the end of the health talk. Socio-demographic characteristics and drinking behaviors were collected at baseline. Students were asked if they agreed on the following 12-items of harm of passive drinking in the pre- and post-questionnaires, including noise; sleep/study interrupted; emotionally hurt; feeling neglected; taking care of a drunk; verbal insulted; physical assault; sexual harassment; unwanted sex; financial loss; accident; and property damaged. Situations of forced drinking , health harm of drinking , and social harm of drinking  were assessed similarly. Intention to quit was measured by asking, “Do you want to stop drinking now?” (yes vs. no), and intention to drink was measured by asking, “Would you drink if your best friend offered you a drink?” (yes vs. no). Both questions were assessed in the pre-and post-questionnaires. The satisfaction level and perceived effectiveness of the Interactive video-based education were measured in the post-questionnaire. t-test was used to measure the change in students’ knowledge of passive drinking , forced drinking , health harm of drinking , and social harm of drinking , after the health talk. McNemar’s chi-squared test was used to assess the change of intention to quit (yes vs. no) and intention to drink (yes vs. no) after the health talk. Sensitivity analyses were conducted for the intention to quit and intention to drink outcomes . The associations of knowledge of passive drinking and forced drinking, with knowledge of drinking harm, intention to drink, and intention to quit, were analyzed using multiple linear regression  and logistic regression (adjusted odds ratio (AOR)), adjusting for sex, school grade levels, perceived affluence, and corresponding pre-talk score. The associations of socio-demographic characteristics with post-talk scores of passive drinking and forced drinking were analyzed using multiple linear regression, mutually adjusting for baseline socio-demographic characteristics and pre-talk scores. Stata 15.1 was used for all analyses. A two-tailed p-value of less than 5% was considered statistically significant. A total of 127 students (9.3%) with missing answers for over half of the questionnaire were excluded. Paired p < 0.001), forced drinking , health harm of drinking , and social harm of drinking , after the health talk. The odds ratio from McNemar’s chi-squared test shows that students were more likely to quit  and less likely to drink alcohol  after the health talk. The differences in the intention to quit  and intention to drink  outcomes, based on the multiple imputed data analyses, were significant.This is the first quasi-experimental study to use interactive video-based intervention to educate adolescents on the harm of passive and forced drinking, to reduce their intention to drink and promote quitting. Our previous survey found that harm was prevalent in Chinese adolescents; 38.9% and 21.9% of secondary students having experienced passive and forced drinking, respectively ,17. Our Traditional school-based alcohol prevention programs showed no significant effects on increasing the knowledge of drinking harm in adolescents . Our stuWe found that the intervention increased adolescents’ knowledge of passive and forced drinking, except for older students (Grade 12) and those who perceived themselves as affluent. Older students were more likely to drink and resort to alcohol to cope with stress and anxiety as they had more external stressors  than younger students . In addiThis study had some limitations. First, the intention to drink and intention to quit were assessed immediately after the health talk, but long-term follow-up data is needed to confirm the effects. The socially desirable answers cannot be excluded. It was less likely given that drinking is prevalent and anonymous questionnaires were used. Second, the causal effects between the intervention and outcomes cannot be concluded, and confounding bias might exist due to the lack of randomization. We have minimized its effects by adjusting for potential confounding variables, and a randomized controlled trial is needed to provide more robust evidence. Third, a students’ experience taking the pre-test questionnaire might affect their performance on the post-test questionnaire (testing effect). We used a 10 min washout period after the health talk to minimize its effect on students’ performance on the post-test questionnaire.We found that our interactive video-based education increased students’ overall knowledge of drinking harm; they also reported less intention to drink after the intervention. Increased knowledge of passive and forced drinking were associated with increased knowledge of drinking harm and less intention to drink. Students who were older and perceived themselves as affluent were not receptive to our intervention; more intensive intervention is needed for these specific groups. This study provides implications that educational programs at school, such as teaching students about the knowledge of passive and forced drinking, can be a feasible strategy to prevent students from underage drinking and developing alcohol misuse problems in the future.