P < 0.05). AI limb rehabilitation system on account of VR technology could effectively recover the daily health management of stroke patients, and its effect was significantly higher than that of traditional drug therapy. This method could be popularized in clinic.This study was aimed at discussing artificial intelligence (AI) limb rehabilitation system on account of virtual reality (VR) on long-term health management of stroke patients. In the study, AI limb rehabilitation system on account of VR technology was compared with traditional drug therapy, and effects of the two therapies on long-term health management of stroke patients were compared. Fifty patients with stroke in the hospital were randomly divided into experimental group and control group with 25 patients in each group. Patients in experimental group were treated with AI limb rehabilitation system on account of VR technology, and those in control group were treated with traditional drug therapy. To compare and judge the recovery of patients' physical ability, patients in the two groups were compared in physical movement ability and daily living activity ability after 10-week treatment. After 10-week treatment, Fugl-Meyer assessment-upper extremity (FMA-UE), Fugl-Meyer assessment-lower extremity (FMA-LE), the Hong Kong version of functional test for the hemiplegic upper extremity (FTHUE-HK), Barthel index (BI) daily living (ADL) activities, and Berg balance scale (BBS) in control group were lower than those in experimental group, but the score of MWS was higher than that of experimental group ( Cerebral stroke is commonly known as stroke in traditional Chinese medicine. Several factors contribute to a cerebral stroke to certain areas of brain tissue, which cause damage to brain tissue . Stroke For patients, stroke is prone to cause disability or apoplexy sequelae . The symUnder the logistics network environment, sensor technology was widely used, which makes the VR technology have further development space in the field of remote control. Current VR technology can provide users with all kinds of training or other types of fun scenarios on account of designs that are closer to real life according to designers, some personalized training time tasks, or personalized learning modes. These learning and training modes that cannot be provided by traditional treatment schemes can provide a novel and interesting rehabilitation scheme for the prognosis of motor dysfunction in stroke patients. At present, the actual effect of VR technology in the long-term health management of stroke patients is not clear.In this study, 50 patients with stroke in the hospital were randomly divided into experimental group and control group. This study innovatively puts forward the artificial intelligence limb rehabilitation system based on virtual reality to train the experimental group, aiming at exploring its advantages in health management of stroke patients.In this study, 50 patients with stroke from May 2019 to December 2020 were selected as research objects. In experimental group, there were 18 males and 7 females, aged 47~68 years old, with an average age of 54.2 ± 13.7 years old and an average onset time of 141.54 ± 38.28 days. There were 21 patients with cerebral infarction, 4 patients with cerebral hemorrhage, 14 patients with upper extremity (UE) hemiplegia, and 11 patients with lower extremity (LE) hemiplegia. In the control group, there were 25 patients with 15 males and 10 females, aged 43~67 years old, with an average age of 56.9 ± 14.6 years old and an average onset time of 132.29 ± 41.62 days. There were 23 patients with UE hemiplegia, 2 patients with cerebral hemorrhage, 13 patients with UE hemiplegia, and 12 patients with LE hemiplegia. This study had been approved by ethics committee of hospital, and patients' families were informed of it and signed informed consents.Key Points of Diagnosis of Various Cerebrovascular Diseases in the 4th National Academic Conference on Cerebrovascular Diseases in 1996, and it was confirmed by brain computed tomography or nuclear magnetic resonance examination [Inclusion criteria were as follows: First, the symptom of patients was in line with mination . Second,Exclusion criteria were as follows: First, patients had serious cardiovascular diseases. Second, patients were not suitable for exercise such as lower limb fracture or surgery. Third, patients had severe cognitive dysfunction and deep sensory impairment. Fourth, patients had myocardial infarction, heart failure, and other diseases recently. Fifth, patients had unstable vital signs and organ failure. Sixth, patients could not stay in the hospital all the time and complete the 10-week rehabilitation program.In the control group, patients received general traditional rehabilitation training methods, and professional medical staff customized personalized rehabilitation training plans according to specific conditions of patients. Specific projects of rehabilitation training were traditional medicine, physical therapy, speech therapy, and occupational therapy (OT). These four specific projects were the traditional treatments for the prognosis of stroke patients. OT, as a kind of training task, mainly provided patients with necessary training tasks through selective operation activities, so as to meet the recovery of various physiological functions. The followings were its main training programs: upper limb fetching exercises, arms forward, and back rotation exercises for shoulder and elbow joints; screwing screws, grabbing objects, and others for finger flexibility; steering training, motion maintenance training, standing balance training, sitting up training, and bridge movement for lower limb joint; and roller training, dressing training, routine living equipment training, and routine walking training which includes walking up and down stairs, lateral walking, cross walking, and others for daily life .In the experimental group, patients received training tasks on account of VR technology in addition to traditional treatments for stroke patients such as traditional medicine, physical therapy, speech therapy, and OT. VR system was divided into motion sensor, motion capture suit, computer monitor, VR digital training system, and training evaluation software. Medical staff wore motion capture suits according to patients' training and physical conditions during general training and fixed motion sensors at the corresponding joints of the body. Motion capture was then performed according to patients' real-time exercise rehabilitation training, transferred to computers through three dimensional (3D) spatial parameters, motion changes, and virtual motion system, and fed back to monitors.At present, VR digital training system mainly consists of multitouch training table and universal treadmill. Multitouch training table is mainly for upper limb training of patients. It has a large number of training modules, including fruit cutting games, driving games, whac-a-mole games, picture combination games, goal tracking games, and coordination training. As the main equipment for limb training, universal treadmill is collected and fed back the walking and running data of patients during rehabilitation training. It can be matched with maze games, balance training, running training, obstacle avoidance, and so on. At the same time, this kind of training module supports the modification of training time and difficulty and combines with the training evaluation software, and the training content can be personalized and adjusted according to the training effect of patients. A training experience integrating interest, creativity, immersion, and interactivity can be created by providing patients with auditory, tactile, and visual sensory stimulation. In this way, patients can be actively immersed in the training content and maximize the training effect. VR training lasts 40~60 min a day, 5 days a week for 10 weeks.The exercise ability of patients in two groups was evaluated prior and post 10-week rehabilitation treatment. Fugl-Meyer assessment-upper extremity (FMA-UE), Fugl-Meyer assessment-lower extremity (FMA-LE), the Hong Kong version of functional test for the hemiplegic upper extremity (FTHUE-HK), Barthel index (BI) of activities of daily living (ADL), Berg balance scale (BBS), and 10 m maximum walking speed (MWS) were used in this study. FMA-UE and FMA-LE belonged to FMA. The mobility of the upper and lower limbs could be assessed by them separately. It could be selected according to hemiplegia patients. The lower the score was, the more severe the injury degree of upper and lower limbs would be. The upper limb function test of hemiplegia mainly aimed at the evaluation of the functional recovery ability of Chinese patients with stroke or hemiplegia in the upper limb rehabilitation training. It had high pertinence and credibility. Patients' upper limb mobility could be graded according to 12 items, and the higher the grade was, the better the upper limb mobility would be. BI scored patients' ADL through a series of independent behavioral evaluations. The program was divided into 10 items which included eating, washing, dressing, going to the toilet, dressing, walking, defecating, urinating, moving utensils, and walking up and down stairs. The higher the score was, the stronger the independent living ability would be. BBS evaluated patients' lower limb balance ability through 14 evaluation criteria. The higher the score of patients was, the better their behavioral ability would be. 10 m MWS mainly aimed at the evaluation of patients' lower limb mobility. It selected a barrier-free route with a straight line distance of 16 m and calibrated it at the starting point, 3 m, 13 m, and the end point. It recorded the time required 3 m~13 m by selecting a barrier-free route with a straight line distance of 16 m. This process was repeated 3 times. Appropriate rest could be carried out in the middle, and the shortest time was selected as the final time –18.P < 0.05 indicated significant differences.Statistical product and service solutions 19.0 version was used for data analyses, measurement data were expressed by P > 0.05), which indicated comparability.The comparison results of specific clinical data statistics of patients were shown in The evaluation tables of prior and post rehabilitation treatment of all patients in two groups were compared, and the specific results were shown in Figures 2P < 0.05).In P < 0.05).In P < 0.05).In P < 0.05).In P < 0.05).In P < 0.05).In According to the above results, AI limb rehabilitation system on account of VR technology had a better therapeutic effect on the prognosis rehabilitation training of stroke patients compared with the traditional rehabilitation training.The behavioral rehabilitation of patients with cerebral apoplexy had always been one of the important problems for patients and their families. After stroke, a large number of patients had hemiplegia, paralysis, loss of self-care ability, and other sequelae. They seriously affect daily lives and bring a lot of inconvenience to themselves and their families. Therefore, it was very important to establish a set of scientific and effective motor rehabilitation management for stroke patients. Therefore, this study intended to build a rehabilitation training system that could not only meet the needs of daily rehabilitation training but also improve patients' training initiative and dependence. In recent years, with the hot and development of VR technology, it was possible to construct 3D space and supporting interactive environment without being affected by environmental factors, which was basically consistent with the requirements of this study. Therefore, AI limb rehabilitation system on account of VR technology in the rehabilitation training of stroke patients was one of the future development trends .The objective of this study was to compare AI limb rehabilitation system on account of VR technology with traditional drug therapy on the rehabilitation of limb movement ability of stroke patients. 50 patients were randomly divided into control group and experimental group and underwent rehabilitation training for 10 weeks, respectively. Compared with traditional treatment, AI limb rehabilitation system on account of VR technology could better improve the rehabilitation efficiency of stroke patients and play an obvious role in the recovery of all physiological motor functions of patients. The reasons could be the followings: first, AR limb rehabilitation technology on account of VR technology could track and analyze the rehabilitation training activities of patients in time, adjust the exercise mode and difficulty in time, take into account the individual needs of users, and avoid a lot of homogeneous meaningless or low-intensity training. Second, it could bring more fun, guarantee a certain novelty, improve the dependability of the rehabilitation training in patients with cerebral apoplexy and initiative, and thus improve the effect of rehabilitation training brought about by the unique immersive VR technology and creativity to users' touch, vision, and hearing stimulated at the same time. Third, it could correct users' trained actions in time with the help of AI in computer systems. It had higher patience and carefulness compared with traditional rehabilitation training, could ease users' mood through sound effects and environment transformation at the same time, and avoid the psychological anxiety caused by a long time training. Fourth, it was easy to operate and could adopt different operating systems according to the users' different modules. At the same time, it was not limited to the requirements of the sites and could move thousands of miles between the square inch through VR technology. Users could not only play the rehabilitation training but also train work and life abilities after rehabilitation by various scenes through the establishment of the corresponding positive feedback mechanism of the system. However, there were also many problems in it: first, the price was too expensive; second, device security should be considered; and third, there were fewer development modules. At present, VR technology was an emerging industry and its supporting software is not perfect, so it still needs further development and improvement in clinical use.Faria et al.  designedThis study was aimed at discussing AR limb rehabilitation system on account of VR technology in the prognosis of stroke patients. The clinical value of it was judged by comparing the rehabilitation effect of the rehabilitation system with that of ordinary stroke patients. The system had a better effect on the prognosis of stroke patients than ordinary training and had a high clinical value. The deficiency of this paper was that the artificial intelligence limb rehabilitation system based on virtual reality technology is expensive and not suitable for large-scale clinical popularization. On the other hand, because the system was a new industry, the types of matching software are not perfect at present. Therefore, although the experiment proves that this method has a high rehabilitation effect, it was still recommended to wait for the development of virtual reality technology. This study provided a new direction for the prognosis and rehabilitation of patients with cerebral apoplexy, the corresponding experimental data, and some clinical experience for the development of technology and clinical use in the future.


Modern evolutionary biology offers a wide variety of methods to explore the evolution of species and to describe their relationships. The methods of DNA/RNA sequence analysis have been developing for decades and have become increasingly popular and reasonably reliable. Nevertheless, final phylogenetic trees for many taxa are still under debate because both classical and genomics-based approaches have their own limitations for phylogenetic tree reconstruction. Here, we propose the use of younger ‘omics’ methods, namely quantitative metabolomics, to aid the phylogeny reconstruction of vertebrates. We show that metabolomics-based hierarchical clustering analysis trees match, although not perfectly, to the genomics-based trees.Milvus migrans), Eurasian magpie (Pica pica), Northern raven (Corvus corax), Eurasian coot (Fulica atra), Godlewski’s bunting (Emberiza godlewskii), Great crested grebe (Podiceps cristatus), Great tit (Parus major), Hawfinch (Coccothraustes coccothraustes), Hooded crow (Corvus cornix), House sparrow (Passer domesticus), Rock dove (Columba livia), Rook (Corvus frugilegus), Short-eared owl (Asio flammeus) and Ural owl . Further analysis shows that the statistical approaches generally used in metabolomics can be applied for differentiation between species, and the most fruitful results were obtained with hierarchical clustering analysis (HCA). We observed the grouping of conspecific samples independently of the sampling place and date. The HCA tree structure supports the key role of genomics in the formation of the lens metabolome, but it also indicates the influence of the species lifestyle. A combination of genomics-based and metabolomics-based phylogeny could potentially resolve arising issues and yield a more reliable tree of life.In the current pilot study, we propose the use of quantitative metabolomics to reconstruct the phylogeny of vertebrates, namely birds. We determined the concentrations of the 67 most abundant metabolites in the eye lenses of the following 14 species from 6 orders of the class Aves (Birds): the Black kite ( To explore the evolution of species and to describe their relationships, traditional taxonomy, for a long time (since Linnaeus and Darwin in the 18–19th centuries), was based on systemically analyzing the traits of species with respect to their morphology, physiology and behavior , which hThe morphological traits of animals are closely connected with biomolecules—nucleic acids, proteins and metabolites—therefore, biomolecules have become useful additional indicators of phylogenetic relationships between species ,4. Molecomics’ methods, such as metabolomics, can contribute to the more reliable construction of phylogenetic trees. Several recent papers have proposed to use the metabolomic approach for the classification of species. This approach has primarily been applied to plant species and subspecies [Today, many scientists are in favor of a more integrative taxonomy ,7,8, andbspecies ,13,14,15bspecies ,17, animbspecies  and humabspecies . Among tbspecies ,16,18,19bspecies ,25,26,27bspecies . Thus, tbspecies . One of The phylogenetic dendrogram is a hypothesis that is often debated for many taxa, because both classical and molecular approaches have their own limitations for relevant interpretation and phylogenetic reconstruction ,27,30,31In the current paper, we propose the use of quantitative metabolomics for the taxonomic differentiation of vertebrates. For the successful application of metabolomics for resolving phylogeny, suitable organs and tissues of organisms should be selected for sampling and further analysis beforehand. Eye lenses seems to be a promising tissue for this purpose. The lens is the transparent tissue of an individual, which is anatomically isolated from other tissues . To be tPica pica), Northern raven (Corvus corax), Hooded crow (Corvus cornix), Rook (Corvus frugilegus), Godlewski’s bunting (Emberiza godlewskii), Great tit (Parus major), Hawfinch (Coccothraustes coccothraustes) and House sparrow (Passer domesticus), belong to the Passeriformes order. Four out of these eight species  correspond to the Corvidae family, and the other four correspond to the Passerides infraorder. Other birds, including the Black kite (Milvus migrans), Eurasian coot (Fulica atra), Great crested grebe (Podiceps cristatus), Rock dove (Columba livia), Short-eared owl (Asio flammeus) and Ural owl , represent non-passerine orders. All species under analysis are rather widespread in Siberia (Russia), and the samples were possible to obtain.This paper is a pilot interdisciplinary study directed at the possibility of using quantitative metabolomics and corresponding conventional unsupervised statistical approaches  for the differentiation between species of the Animalia kingdom in comparison with their phylogenetic relationships . To thiAt the molecular level, the adaptations of species are imprinted in all layers of the functional organization of life: the genome, transcriptome, proteome and metabolome. Among these layers, the genome is the most conservative, whereas the metabolome is the most susceptible to the variability of many external and internal factors. Consequently, the composition and the concentrations of metabolites in the lens, or the lens metabolome, are determined by the combination of conservative  and variable  factors and thus is expected to vary to some extent between species. It is interesting to establish the magnitude of such variations and to evaluate the contributions of conservative and variable factors into the formation of the total metabolome of the lens.The major goals of the current study are: (1) to identify the most abundant metabolites in the lenses of 14 bird species from 6 orders and to establish their concentrations; (2) to analyze the metabolomic composition of bird lenses; (3) to assess the applicability of the generally used statistical metabolomic approaches to the phylogenetic differentiation between species and to compare the genomics- and metabolomics-based trees; and (4) to evaluate the influence of conservative and variable factors on the total metabolome of the lens.2O 99.9% was purchased from Armar Chemicals . Acetonitrile HPLC grade of quality 0 was purchased from Cryochrom . All other chemicals were of the highest purity, usually HPLC or LC-MS grade, and were purchased from Sigma-Aldrich . H2O was deionized using an Ultra Clear UV plus TM water system  to the quality of 18.2 MOhm.Chloroform and methanol HPLC grade were purchased from Panreac . DThe study was conducted in accordance with the ARVO Statement for the Use of Animals in Ophthalmic and Vision Research and the European Union Directive 2010/63/EU on the protection of animals used for scientific purposes, and with the ethical approval from the International Tomography Center SB RAS (ECITC-2017-02).The bird species were collected from three sources : (1) durThe species under analysis are common in Siberia, and the samples were relatively accessible. All species were adults of wild origin and were obtained within the 2017–2020 time period. Lenses were extracted clean from the bodies, placed into separate cryotubes, frozen in liquid nitrogen and kept at −70 °C until the analysis within the 2017–2021 time period. Depending on the size of a lens, each sample contained one to three lenses from different individuals .g, +4 °C for 30 min, yielding two immiscible liquid layers separated by a protein layer. The upper aqueous layer (MeOH-H2O) was collected, divided into two parts for NMR (2/3) and LC–MS (1/3) analyses and lyophilized.The lens sample preparation was performed by the procedure that has been described in detail ,39,40. W2O containing 2 × 10−5 M sodium 4,4-dimethyl-4-silapentane-1-sulfonic acid (DSS) as an internal standard and 50 mM deuterated phosphate buffer to maintain a pH of 7.2. The 1H NMR measurements were carried out at the Center of Collective Use “Mass spectrometric investigations” SB RAS with the use of an NMR spectrometer AVANCE III HD 700 MHz  equipped with a 16.44 Tesla Ascend cryomagnet, as described in [The extracts for the NMR measurements were re-dissolved in 600 μL of Dribed in . The con2O. The LC separation was performed with an UltiMate 3000RS chromatograph  using hydrophilic interaction liquid chromatography (HILIC) on a TSKgel Amide-80 HR  column . The MS detection was performed with an ESI-Q-TOF high-resolution hybrid mass spectrometer maXis 4G , as described earlier [In this work, LC–MS data were used only for the identification of several unknown compounds and for the confirmation of the data obtained by the NMR method. The extracts for the LC–MS analysis were re-dissolved in 100 μL of 10 mM ammonium formate and 0.1% formic acid solution in H earlier ,40.www.metaboanalyst.ca (accessed on 10 January 2022) [The statistical treatment of the quantitative metabolomics data—principal component analysis (PCA), hierarchical cluster analysis (HCA) and heatmap construction—was performed at the MetaboAnalyst 5.0 web-platform (ry 2022) ) with thPython/R statistical packages allow for choosing a wider variety of HCA parameter values . However, the current pilot work is aimed at assessing the general applicability of the quantitative metabolomics-based HCA method for phylogeny. Thereby, we decided not to go deep into developing the correct HCA parameters, and thus we used only the built-in MetaboAnalyst ones. Moreover, we did not restrict possible combinations of values. The general guideline for choosing the appropriate HCA parameters is to use ones that maximize the distance between samples in different classes and to minimize that within each class.For the construction of dendrograms based on hierarchical clustering, several parameters related to the HCA data treatment need to be chosen: the method of the data normalization , the type of the distance\similarity measure , and the clustering algorithm . The HCA parameter values listed above are available as built-in options at the widely-used-in-metabolomics web platform MetaboAnalyst v5.0, where the current analysis was performed. It should be noted that other tree-building software or the Passeriformes includes 8 out of 14 species under investigation, and a more detailed tree for this order was adapted from the work of Oliveros CH et al. [In a preliminary analysis of the metabolomics-based dendrograms, we observed similarity in the topology with the published phylogenetic dendrograms based on different approaches. To test this hypothesis and to compare the metabolomics-based trees and the “classical” genomics- or transcriptomics-based phylogeny, we manually reconstructed two phylogenetic trees from the literature to fit only the 14 bird species under study. The backbone parts of a tree from the class (Aves) to the orders  were based on two recently proposed modern bird phylogenies, either on work of Jarvis ED et al.  or on thH et al. . To thisThe identification of metabolites was performed according to their NMR spectra that are available in the literature, in databases  and in our in-house NMR library ,35,39. IA. flammeus and S. uralensis, the measurements were performed for 3–12 individuals  are collected in The concentrations of the metabolites in the lenses (in nmol/g) were calculated by the integration of the NMR signals relative to the internal standard DSS followed by normalization to the tissue wet weight. Typically, 60–80 compounds were identified for every species; however, the NMR signals from some compounds were either too weak or strongly overlapped by other signals, which made the quantification of these compounds unreliable. For that reason, the final set of metabolites studied in this work was restricted to 67 identified compounds and 7 unknowns. For every species, except ividuals , and themyo-inositol and taurine. In the lens of A. flammeus, the concentration of lactate was also slightly above 20 µmol/g. Both myo-inositol and taurine are well-known osmolytes [P. cristatus, S. uralensis and A. flammeus, both osmolytes share the function of osmotic protection and have very high abundance above 10 µmol/g ; in P. cristatus, myo-inositol prevails tenfold over taurine. In all four Passerides , taurine has a higher abundance than myo-inositol (up to 1.9 times), and in both Strigidae , taurine prevails 5–10 times over myo-inositol.There were two principal metabolites in the bird lenses with an average concentration above 20 µmol/g: smolytes ,35,43,440 µmol/g . In the myo-inositol, taurine, glutamine and serine); antioxidant protection (glutathione and ergothioneine); and cellular energy generation .Eight further metabolites had average concentrations below 10 and above 3 µmol/g, namely, lactate, glutamine, acetate, glutathione, alanine, ergothioneine, serine and ATP. These compounds can be ascribed to the major metabolites of bird lenses . High leP. domesticus); proline and betaine (E. godlewskii); methionine and anserine (F. atra); leucine and D139 (P. cristatus); UDP (A. flammeus); and NADH (M. migrans).There were six metabolites with concentrations above 1 µmol/g in more than one species : glutamaA. flammeus and S. uralensis, and for P. cristatus  may start to vanish in the PC1 vs. PC2 plot. In this case, the search for the discrimination between groups can include the use of additional dimensions with subsequent principal components  or reductions in the number of groups by removing distant groups. In the present work, we performed PCA for 67 samples from 14 groups . It is worth noting that, independently of the sampling place and date , and Passerides and the Corvides samples form a larger Passeriformes cluster. Similar to the PCA analysis, the grouping of the conspecific samples is observed independently of other factors, such as age, sex or sampling place and date , is demonstrated in P. cristatus species is the most correct compared to the other HCA trees, and the samples are the most distant from other clusters.Data normalization  is often required to equalize the contributions of high- and low-abundant metabolites into analysis; if one uses non-scaled data, the position of the species in the dendrogram is determined by the concentration variation for the several most abundant metabolites only. The exception from this rule is the use of non-scaled data with nonparametric statistics , since the latter discards the information on the concentrations. On the other hand, taking into account the information on metabolite abundance in some specific form can potentially be helpful for phylogeny. For example, for PSS clustering , the posP. cristatus differ significantly from other species. For C. coccothraustes, two unknowns (S112 and S120) are flaring. For both Strigidae species, 10–11 metabolites  comparison of the relative content of metabolites in different groups of samples. Most often, semi-quantitative data are obtained with the use of electrospray ionization-based mass spectrometry (ESI-MS). The ESI-MS signal intensity is not directly proportional to the metabolite concentration; metabolites with a low concentration but a high ionization ability can give more intense signals than metabolites with a high concentration but a low ionization ability. In addition, matrix effects and ionization suppression can lead to the biased interpretation of data ,46.1H NMR) is often used. Peak areas of the metabolite signals in NMR spectra are directly proportional to the compound concentrations, which makes the determination of the metabolite levels in a tissue easy and straightforward. The results of the present work, obtained with the use of NMR-based quantitative metabolomics, demonstrate that the data obtained can be used not only for the comparison of the biological features of different species, but also for animal classification and for studying the evolution of biochemical processes in living nature.The quantitative approach yields absolute concentrations of metabolites in a tissue . It demands much more effort and time than qualitative or semi-quantitative methods, but the data obtained have a long-term value. In addition, the results of such experiments are available for future ‘eternal’ re-use, such as for data mining, new interpretations, the addition of new samples or groups, new comparisons, etc. . That camyo-inositol and taurine, which share the function of lens osmotic protection [R. norvegicus), contain high concentrations of taurine, which prevails almost tenfold over myo-inositol (13.2 and 1.8 µmol/g correspondingly) [myo-inositol in the eye lens may act as a chaperone, protecting the lens proteins from the aggregation caused by post-translational modifications during a lifespan. Most likely, low levels of myo-inositol in owl lenses should be attributed to their nocturnal lifestyle, so the light-induced protein modifications for these species are less dangerous.We have found that the composition and concentrations of the most abundant metabolites vary from species to species, but their levels in genetically close species are very similar, which indicates the importance of the genetic factor in the formation of the tissue metabolome. The eye lenses of all birds contain very high concentrations of two metabolites, otection ,34,43,44ndingly) . It has ndingly)  that, bemyo-inositol, taurine, lactate, glutamine, acetate, glutathione, alanine, ergothioneine, serine, ATP, glutamate, creatine, pyroglutamate, glucose, phosphocholine, valine, ADP, glycine, proline, betaine, methionine, anserine, leucine, UDP and NADH), which emphasizes their significance in important biochemical processes that are genetically adapted to the lifestyle necessities of the species. These high-abundant metabolites are involved in osmotic protection, antioxidant protection and cellular energy generation . It should be noted that a metabolite usually shares several functions and participates in several pathways; thereby, the metabolite role is not limited to the given functions and pathways.A number of quantified metabolites in bird lenses, on average, have rather high concentrations  genetically close bird species have similar concentrations of the most abundant metabolites ; (B) in Conspecific samples are positioned together and form clusters separated from other species .C. coccothraustes, E. godlewskii, P. domesticus, P. major) are positioned together in a separate larger cluster.All four Passerides  are positioned together in a single cluster.All four Corvides .M. migrans, A. flammeus, and S. uralensis), either from the Afroaves clade [Birds of prey ; no node between F. atra and C. livia  was fouC. frugilegus and P. pica are well clustered with the Corvides, they have rather incorrect phylogenetic distances within the Corvides. C. frugilegus should be closer to the other species from the Corvus genus (C. corax and C. cornix), and P. pica should be the sister taxon to all Corvus. The samples of P. pica, C. corax and C. cornix often mix together, without the formation of separate clusters for each species .Although P. major; it should be more distant from the other species of the Passerida parvorder (light green arrow).Similarly, the incorrect positioning is observed for The following differences between the HCA and phylogenetic trees ). The most fruitful results were obtained with the hierarchical clustering analysis. The topology of the obtained dendrograms is very similar to the topology of genetics-based trees, and general phylogenetic dependencies are well-reproduced, although none of the HCA dendrograms ideally fit to the genetics-based trees. The HCA dendrogram structure supports the key role of genomics in the formation of the lens metabolome, but it also indicates the influence of lifestyle. Very likely, most discrepancies in the HCA dendrograms as compared to genomics-based phylogenetic trees originate from different lifestyles and feedings.In the current paper, we applied methods of quantitative metabolomics and corresponding statistical approaches for the differentiation between 14 species from 6 orders of the class Aves (Birds) and for further phylometabolomic tree construction. We determined the concentrations of the most abundant metabolites in the eye lenses of the species and deposited the corresponding raw NMR spectra and the metabolomic analysis into our Animal Metabolite Database repository (Perhaps, the addition of metabolomic data for a larger number of bird species into the analysis, as well as the development of HCA methods, can produce a clearer correlation and similarity of the phylogeny of birds with eye lens metabolomics-based hierarchical clustering. A combination of phylogenetic and phylometabolomic analyses can potentially solve issues in the reconstruction of bird phylogeny and can yield a more reliable tree of life. Current methods can be applied to differentiate other species of vertebrates, and with several adaptations to other species of the animal kingdom and even of other kingdoms, the only limitation is that the tissue under analysis is conservative and comparable between all species under analysis.


Via combined differential gene expression analyses, 6 hub genes were identified: PDGFRA, WNT5A, PPP2R1A, BMPR2, BMPR1A, and SMAD1. This study provides a foundation for further research on the mechanism by which melatonin regulates cashmere growth.The growth of secondary hair follicles in cashmere goats follows a seasonal cycle. Melatonin can regulate the cycle of cashmere growth. In this study, melatonin was implanted into live cashmere goats. After skin samples were collected, transcriptome sequencing and histological section observation were performed, and weighted gene co-expression network analysis (WGCNA) was used to identify key genes and establish an interaction network. A total of 14 co-expression modules were defined by WGCNA, and combined with previous analysis results, it was found that the blue module was related to the cycle of cashmere growth after melatonin implantation. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the first initiation of exogenous melatonin-mediated cashmere development was related mainly to the signaling pathway regulating stem cell pluripotency and to the Hippo, TGF-beta and MAPK signaling pathways.  The Inner Mongolia cashmere goat is an economically important animal with diverse uses and is an important economic breeding stock in China. Cashmere is one of the finest and lightest fibers produced by animals; it is noted for its outstanding properties and rarity and is much more expensive than wool of the same fineness. It is used especially in the production of high-end textiles. Cashmere is produced by hair follicles, an accessory organ of the skin , 2. CashWNT10B, β-catenin and other proteins in the skin tissue of Inner Mongolia cashmere goats. With the rapid development of high-throughput sequencing technology, through the analysis of differentially expressed genes (DEGs) and functional enrichment analysis, scientists have identified many regulatory factors and signaling pathways that may influence the hair follicle cycle: the WNT signaling pathway, fibroblast growth factor (FGF) family, bone morphogenetic protein (BMP) family, Sonic hedgehog (SHH) signaling pathway, transforming growth factor (TGF) family, Notch signaling pathway, etc. , nutrition, genetics, endocrine status, etc. . Studiesay, etc. –10.WNT10A is a key gene in the early stage of development and maturation of fetal skin hair follicles in Inner Mongolia cashmere goats  staining kit  according to the instructions.Total RNA was extracted from the skin of three cashmere goats using an RNAiso Plus Kit (TRIzol method). The purity and integrity of total RNA were measured using a sterile UV–VIS spectrophotometer and an Agilent 2,100 bioanalyzer, respectively, and the three RNA samples were then mixed. Total RNA was stored in a freezer at −80°C.The cDNA library for transcriptome sequencing was constructed according to the operating instructions of an Illumina TruSeqTM RNA Sample Preparation Kit. Total RNA extracted from three cashmere goats per group in each month was mixed together in equal amounts. The mRNA was purified with oligo(dT) magnetic beads and sheared into 100–400 bp mRNA fragments. Double-stranded cDNA was synthesized using the mRNA fragments as templates, an exonuclease and a polymerase. To obtain the sequencing library, the double-stranded cDNA was phosphorylated, ligated to sequencing adapters and subjected to poly(A) tailing prior to PCR amplification.Capra hircus) genome. The FPKM value of each gene was calculated to estimate the gene expression level.The Illumina HiSeqTM 2000 sequencing platform was used for paired-end sequencing of cDNA. A 2 × 100 bp sequencing protocol was used for sequencing by Shanghai Meggie Biopharmaceutical Co., Ltd. After sequencing, referenced and unreferenced genomes were used to compare the reads, and assembly was conducted with the Trinity and Velvet methods. The filtered reads were aligned to the goat |). To remove the influence of low correlation coefficients, an appropriate threshold (β) was selected for exponential weighting of the correlation coefficient matrix and construction of the adjacency matrix. Then, the adjacency matrix was transformed into the topological overlap matrix (dij = 1−TOMij) was then constructed, and hierarchical clustering was carried out for this matrix. The dynamic cutting tool in WGCNA was used to prune the cluster tree, and the gene modules were preliminarily divided. Based on the similarity of the eigengenes of the original modules, these modules were combined to form the final modules.Before WGCNA, the selected gene set was screened and filtered to remove low-quality genes or samples with unstable effects on the results to improve the accuracy of network construction. A weighted gene co-expression network was constructed using WGCNA (V. 147) package in R software . The PeaAs shown in our previous publication , the cycP < 0.05. The goat database was selected for KEGG pathway enrichment analysis with KOBAS 3.0 /Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on the differentially expressed genes between the implanted group and the control group in May. GO enrichment analysis was performed using the online tool g: Profiler . The GO OBAS 3.0 , and P <P < 0.05. Then, Venny 2.1.0 was used to determine the overlap among the three sets of genes.To identify the genes expressed in May that play a role in activating cashmere growth in advance of the seasonal cycle, we screened for the genes differentially expressed in May between the implantation group and the control group. The thresholds for differential gene expression were | log2-fold change | > 1 and PDGFRA, WNT5A, BMPR2, and BMPR1A genes published by NCBI, specific primers were designed with Primer 5.0 (Table 3) and synthesized by Shanghai Bioengineering Co., Ltd. Finally, a TB Green “Premix Ex Taq” II Kit  was used for qRT–PCR. Six samples were tested for each month, and three technical replicates were performed.In this study, an Applied Biosystems QuantStudio 3 real-time fluorescence quantitative PCR system was used to reverse transcribe the extracted total RNA into cDNA according to the instructions of a PrimeScript™ RT Master Mix Kit . Then, based on the cDNA sequences of the goat −△△CT method was used for the analysis of all qRT–PCR data, and SPSS software (version 22.0) was used for statistical analysis. Values are expressed as the means ± standard deviations. A significance level of 0.05 was used.The 2P < 0.01).The S:P (ratio of secondary hair follicles to primary hair follicles) was used as an indicator of the number of hair follicles in the skin of the cashmere goats. In this study, we performed histological staining of sample sections from the control group and the implanted group in May , and sixThe sequencing data for the 24 samples are shown in RNA-seq was performed on 12-month transcriptome data for goat skin samples from the implantation group and the control group. Average linkage hierarchical clustering based on the computed topological overlap was then used to identify genes with very similar expression patterns in each module. A total of 14 co-expression modules were identified in the WGCNA network and represented by different colors. The resulting blue, cyan, green, green–yellow, light cyan, light green, magenta, midnight blue, pink, purple, salmon, tan, turquoise, and yellow modules contained 4,530, 84, 291, 117, 61, 41, 402, 76, 212, 119, 103, 105, 4,574, and 301 genes, respectively  . In the As shown in We further performed GO/KEGG enrichment analyses with 316 differentially expressed genes upregulated in the blue module in May. According to GO classification statistics, 148 terms were grouped into three GO categories: cellular component, molecular function, and biological process. Among the three GO categories, the most significantly enriched were cellular protein modification process, desmosome and protein binding. Notably, the cellular components were significantly enriched in desmosomes, which are the most important intercellular adhesion junctions, directly adhering to desmosomal cadherins on adjacent keratinocytes to form the epidermal layer.PDGFRA, WNT5A, BMPR1A, BMPR2, PPP2R1A, and SMAD1  between the control group and the implanted group in May . Studies have shown that PDGF signaling is related to hair follicle morphogenesis in early hair follicle development and that inhibition of the expression of PDGF or its receptor PDGFRA or PDGFRB in growing and developing hair follicles prevents hair follicle maturation  is an important regulator of the cell cycle and plays a role to similar CDK4 as a mediator of keratinocyte proliferation  or nerve growth factor (NGF) receptors to the Ras/MAPK signaling pathway  gene is to promote DNA synthesis in cells. PDGF participates in the regulation of hair follicle growth and development mainly through PDGF signaling , one of the four major serine/threonine protein phosphatases. The mechanism of this gene in hair follicle development in cashmere goats has not been reported, but based on the signaling pathways and increased expression level of PPP2R1A, we speculate that PPP2R1A may be related to cell proliferation and differentiation, a possibility that will constitute our future research direction. In general, in this study, by using WGCNA, we identified 6 key genes that may mediate the effects of MT and lead to early cashmere production in cashmere goats, providing a foundation for subsequent research.Mammalian hair follicle development begins in the embryonic stage, and studies have shown that Wnt/β-catenin, TGF-beta/Smad and other signaling pathways are involved in the early initiation, differentiation and development of the hair follicle tissue structure . In skinPDGFRA was identified as the key gene through which MT regulates cashmere growth, providing a theoretical basis for further improving cashmere yield and economic benefit.In this study, WGCNA was used to investigate the mechanism by which exogenous melatonin regulates the cycle of cashmere developmental. Histological sections of goat skin that were collected in May showed that the hair follicles of the control group were located deeply in the epidermal layer, while the hair follicles of the implanted group were located near the epidermal layer. The ratio of secondary hair follicles to primary hair follicles in the implanted group was significantly higher than that in the control group. A total of 14 co-expression modules were identified, and the blue module was found to be correlated with the cashmere growth cycle after MT implantation. Through analysis of the hub genes, the genes in the blue module and the genes differentially expressed between the implanted group and the control group in May, https://www.ncbi.nlm.nih.gov/, SRP145408.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: In this study, skin samples were collected in accordance with the International Guiding Principles for Biomedical Research involving animals, and the experiment was approved by the Special Committee on Scientific Research and Academic Ethics of Inner Mongolia Agricultural University, which is responsible for the approval of Biomedical Research Ethics of Inner Mongolia Agricultural University [Approval No. (2020) 056]. No specific permissions were required for these activities, and no endangered or protected species were involved .ZhihL and YZhao contributed to the concept and design of the study. ZhicL, MZ, TC, ZhixW, CZhao, CZhan, QQ, XX, and ML collected the samples and extracted the RNA. QM performed the sections and staining. ZhicL, YX, and QQ conducted the data analysis. YZhan, RS, ZhiyW, RW, and JL provided technical support. ZhicL and ZhihL wrote the first draft of the manuscript. All the authors contributed to the revision of the manuscript and read and approved the submitted version.This work was supported by the National Key R&D Program of China (2021YFD200901); the National Natural Science Foundation of China ; the Major Science and Technology Projects of Inner Mongolia Autonomous Region (2020ZD0004); the Key Technology Project of Inner Mongolia Autonomous Region (2020GG0030 and 2020GG0031).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Sassafras tzumu stands during the self-thinning phase. Between the ages of 10 and 40 years, the number of stems per hectare has declined from 1,068 to 964 due to density-dependent mortality. As the relative stand density decreased, there were significant reductions in the average tree ring width (5.07–3.51 mm) and increases in latewood proportions (49.88–53.49%) and the density of the annual growth ring (165.60–708.58 kg/m3). Therefore, ring density, earlywood density, and latewood density increased with decreasing relative stand density after self-thinning occurred. Ring width, earlywood width, and latewood width significantly increased from the base to the apex of the stem. Stand density and stem height had additive effects on S. tzumu wood properties during the self-thinning phase. A shift in the growth allocation along the longitudinal stem in response to self-thinning resulted in decreasing radial growth, increasing wood density, and improved stem form. In summary, we found a significant influence of stand density on tree ring growth, wood quality, and stem form of S. tzumu trees during the self-thinning phase.Silvicultural practices greatly improve the economic value of wood products from forests. Stem dimensions, wood density, and stem form are closely linked to end-product performance. This research aimed to examine the effects of stand density and stem height on variables that reflect ring growth and wood properties of  Silvicultural practices that affect tree growth may cause changes in the economic value of the raw material. Of the different intensive management treatments, stand density management is of critical importance to end-product properties . For exaTsuga heterophylla, Picea glauca, and Picea engelmannii have examined tree growth and wood quality responses to competition, these studies focused on a specific tree height   . Given t 1.37 m) , it is iSassafras tzumu is one of the most important broad-leaved deciduous tree species in southern China  investigate the influence of relative stand density and longitudinal stem position on diameter increment, wood density, and stem form; (ii) assess growth allocation along the stem from the stem base toward the stem apex; and (iii) detect whether there are interactive effects of stand density and longitudinal stem position on wood properties within each growth ring.Therefore, this study aimed to examine the effects of stand density and longitudinal stem height on variables that reflect ring growth and wood properties of 3/ha/year at the age of 20 years. The intermediate quality site was located at middle slope and had a mean annual increment of 11.7 m3/ha/year at the age of 20 years. The low-quality site was located at upper slope and had a mean annual increment of 8.5 m3/ha/year at the age of 20 years.The study was conducted in the Huangshan Forestry Teaching Farm , in Mt. Huangshan, Anhui Province, China . The cliS. tzumu plots on different sites were established in 1977 at a planting spacing of 2.0 m × 3.0 m . The plot size was 20 m × 30 m (ten rows with ten trees) with a buffer zone consisting of 3 rows at the same spacing. This initial spacing was chosen because it is the most used planting spacing for the establishment of S. tzumu plantations.Six V, m3) was estimated using the formula V = 5.0479055 × 10–5 × (DBH)1.9085054 × (TH)0.99076507 , and diameter at breast height  were measured for all trees and the stem volume , at the following heights: stem base , breast height (1.30 m), 1/4 TH, 1/2 TH, and 3/4 TH.3), earlywood density , and latewood density . These variables were measured using a high-frequency densitometry measurement LignoStation™ system  with 1/1,000 mm along four radii taken 90° from each other, with the initial radius selected at random , earlywood width , latewood width , latewood proportion , ring density  were used to assess stem taper , butt taper , and the ratio of the diameter at different sampling heights to the diameter at breast height (em taper . ST was Tree-ring indices were used to discriminate between juvenile and mature wood based on the following five steps: (1) the age-related ring growth trends were quantified ; (2) thedi/DBH), following the three-step model selection approach , wood density , and stem form , the dependent variable YANOVA and pairwise Pearson correlation analyses were carried out between all of the variables to determine the significance and strength of their relationships among the different tree properties. All tests were performed at a 5% significance level and the statistical analyses were conducted using SAS software .S. tzumu plots. Relative stand density had an inverse effect on LWP, which increased by 7.2% with decreasing living stems per hectare (Relative stand density significantly and strongly affected RW, EWW, LWW, and LWP . There w hectare .F-values showed that the influence of stem height on these variables was considerably lower than that of relative stand density and cambial age (Ring width, EWW, and LWW changed significantly from the stem base toward the stem apex . There wbial age .Along with the tree ring growth from the pith to the bark, RW showed a tendency to increase rapidly during the early growth stage and then subsequently decrease . The camRelative stand density significantly influenced RD, EWD, and LWD . The RD di/DBH (di/DBH increased (Relative stand density significantly and strongly influenced ST and BT, but it did not have any significant effect on di/DBH . A decredi/DBH , while Bncreased .Regarding the radial variation of RD and LWD along with the radial disc, RD increased from the pith outwards until the maximum value was reached around 3–7 years, and then decreased with increasing cambial age in mature wood .di/DBH (Ring width was positively correlated with EWW, LWW, and ST, and it was negatively correlated with RD, EWD, LWD, and BT. There was a significantly negative correlation between EWW and LWW. RD exhibited highly significant, positive correlations with EWD and LWD, and LWD was negatively correlated with di/DBH .S. tzumu plots density decreased, there were improvements in wood quality and tree form. We found a strong negative correlation between ring growth and wood density of the even-aged pure S. tzumu stands during the self-thinning phase. It is worth mentioning that these findings contrast with several other studies on conifer, ring-porous hardwood, or diffuse-porous hardwood species (This study showed the patterns of tree ring growth, wood density, and stem form along a stand self-thinning trajectory. Our findings indicated that, as the  species . For exa species . A posit species . In diff species .S. tzumu plantations, the relative stand density was positively correlated with ring width and negatively correlated with tree ring wood density. In this way, with the decrease in relative stand density because of competition-induced mortality, there was a decrease in ring width and an improvement in wood density due to greater mature wood formation initiating approximately 17 years after planting. This contrasts with other studies in which relative stand density was positively correlated with tree ring wood density and negatively correlated with tree ring width (For ng width , which mng width .Larix kaempferi, such that high growth rates resulted in lower-density wood during the juvenile period (As the cambium cells become mature, the tendency is to reduce the width of the growth ring and to produce mature wood, which is characterized by a higher LWP compared with juvenile wood . This hae period . With the period .S. tzumu, the juvenile wood was produced during the first 17 years. This indicates that when relative stand density exceeded 1.07, between the ages of 10 and 20 years, there was a predominance of juvenile wood being formed. On the contrary, as the relative stand densities declined, which occurred at the ages of 30 and 40 years, there was a predominance of mature wood formation that is characterized by an increase in LWP, which is of a higher density. For Picea mariana, the simultaneous variation of juvenile wood diameter and tree diameter along the stem direction leads to a constant proportion of juvenile wood (S. tzumu during the self-thinning phase. The results indicate that there was an increase in the diameter of the tree and the formation of mature wood with significant improvements in the quality of S. tzumu wood during the self-thinning phase as RD increases, especially after the age of 17 years.Part of the effect of stand density is probably also the effect of stand age. As the cambium cells become mature, there is a trend of decreasing ring width and increasing LWP, leading to mature wood formation . For S. ile wood . These aCompetition was maintained at high levels during the self-thinning phase, and RD decreased along the axial stem direction from breast height to the stem apex. This was related to increasing EWW and decreasing LWW along the stem from the base to the apex. This trend in RD along the stem probably results from the higher proportion of juvenile wood at the stem apex compared with that at lower stem positions along the stem .S. tzumu is a shade-intolerant deep-rooting tree species, and the rapid growth of S. tzumu individuals can enhance its access to light and outcompete the slow growth of neighboring trees.The effects of stem height were often significant on stem dimension and stem form. The axial variation at a specific stem height proceeds downward and upward along a stem as a stand undergoes self-thinning , which eS. tzumu plantations.In summary, as relative stand density decreased throughout the self-thinning phase, wood density and LWP increased, whereas growth ring width and stem taper decreased. From a silvicultural perspective, we showed that, in conditions similar to those in our study, one can expect the predominant formation of mature wood to start at about 17 years after planting. Our analysis of wood properties shows the importance of considering the position along the stem when examining wood properties. Such information is valuable for the improvement of the silviculture and wood utilization of S. tzumu stands, ring growth values of RW, EWW, and LWW decreased, but LWP increased; RD, EWD, and LWD increased; ST decreased, while BT and di/DBH increased. Decreased tree growth during self-thinning trajectory was offset by improved wood density and stem form. Tree ring growth closer to the apex of the stem and the lower parts of the stem decreased the tree ring wood density during stand self-thinning trajectory. The shift in the internal resource allocation along the stem might prefer to maximize end-product value of timber.During the self-thinning of the even-aged pure This study highlights the important influence of stand density on tree ring growth, wood density, and stem form. A key finding was that the stem height did not significantly interact with stand density for any of the key wood properties during the self-thinning phase, which suggests the additive effects of stand density and stem height have on wood properties.The raw/processed data required to reproduce these findings cannot be shared at this time as the data also forms part of an ongoing study. Further inquiries can be directed to the corresponding author.HS and JJ conceived and designed the experiment. SD performed the experiment and collected the data. HS analyzed the data and wrote the initial draft of the manuscript. AS, TP, and DF edited and revised the final version of the manuscript. All authors approved the final version for publication and agreed to be held accountable for the content therein.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The purpose of this study is to make a systematic review of the therapeutic effect of stem cells in animal models of disc degeneration from an imaging point of view.  Data were extracted by searching electronic databases for RCTs that met the inclusion criteria. Data analysis was performed using RevMan 5.3 and STATA 15.1 software. This meta-analysis was registered with INPLASY, registration number INPLASY202240148. P < 0.001), mice (P < 0.001), sheep (P < 0.001), and rats (P = 0.001) was better than that in control group. In terms of disc height, the efficacy of stem cell group in rats (P < 0.001) was better than that in control group, while in sheep (P = 0.355), there was no statistical difference between two groups. In terms of MRI index, the efficacy of stem cell group in rats (P < 0.001), mice (P < 0.001), and rabbits (P = 0.016) was better than that in control group. In terms of MRI signal score, the efficacy of stem cell group in rabbits (P < 0.001) was better than that of control group. In terms of T2 signal intensity, stem cell group was more effective than control group in rabbits (P < 0.001), mice (P < 0.001), and rats (P = 0.003). A total of 34 studies were included, covering four species of animals, rabbits, sheep, rats, and mice, with a total of 1163 intervertebral discs. In terms of DHI, the efficacy of stem cell group in rabbits ( Stem cell therapy can improve intervertebral disc-related imaging parameters in animal models of disc degeneration, indicating that stem cell therapy has a repairing effect on intervertebral discs. However, given the heterogeneity and limitations of this study, this conclusion still needs to be tested by a large number of studies. Degenerative disc disease is a clinical syndrome caused by permanent structural changes in the intervertebral disc, and it is one of the most common clinical diagnoses . It has Disc degeneration is a complex multifactorial process determined by genetic, nutritional, and mechanical factors . It is cIn the past 20 years, people have tried to find new breakthroughs in the treatment of disc degeneration from stem cells. Encouragingly, these efforts are paying off. There has been a growing body of research demonstrating the therapeutic potential of stem cells in animal models, and clinical studies are taking the first steps , 8–10. SThis study was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement  and the Studies that met the following requirements were included: (1) randomized controlled trials (RCTs) using animal models of disc degeneration as the research object, and the animal species were not limited; (2) stem cell therapy was used as an intervention measure, and other drugs were used as control measure. Both could be combined with other drugs or materials; (3) there were no restrictions on the source of stem cells, the dose of drugs, and the course of treatment.Studies were excluded according to the following exclusion criteria: (1) the animal species used were different from other literatures and could not be compared with any of the other included studies; (2) the imaging evaluation indexes or imaging data representations used were different from other literatures and could not be compared with any other included studies.After determining the inclusion and exclusion criteria for this study, two researchers independently searched multiple databases, including PubMed, Cochrane Library, ScienceDirect, CNKI, and Wanfang Database. The retrieved articles were published before March 1, 2022. The following search terms were used: disc degeneration, animal model, stem cell, mesenchymal stem cell, bone marrow-derived mesenchymal stem cell, adipose-derived stem cell, MSC, BMSC, and ADSC with the Boolean operators AND or OR. The retrieved literatures were screened by two researchers step by step according to title, abstract, and full text. After identifying included articles, we traced their references to identify potential articles.After screening was complete, data were extracted from eligible literatures by two independent researchers and then cross-checked by a third researcher. For the differences in the included literatures, all researchers reached consensuses through discussion. The data extracted in this study included the name of the first author, year of publication, country, animal species, modeling method, stem cell type, stem cell source, injection dose, interventions in the control group, and imaging evaluation indicators.Since the target literatures for this study were RCTs in animals, we used the Cochrane Risk Bias Tool  for qualI2 statistic. The greater the I2, the greater the heterogeneity. If there was heterogeneity in this study (I2≥50%), a random-effects model was used; otherwise, a fixed-effects model (I2<50%) was used. In this study, differences were considered statistically significant when P < 0.05.We performed statistical analysis of data extracted from each study using STATA software (version 15.1). Continuous variables were reported as mean difference (MD) and 95% confidence interval (CI), while dichotomous variables were reported as odds ratio (OR) and 95% CI. Statistical heterogeneity was judged according to the n = 136), Cochrane Library (n = 2), ScienceDirect (n = 354), CNKI (n = 400), and Wanfang Database (n = 86). After removing duplicate studies, 34 studies were finally included according to the inclusion and exclusion criteria. The flow chart of literature screening is shown in According to the above search strategies, 978 relevant articles were preliminarily identified, including PubMed , so a random-effects model was used. The comprehensive results showed that the difference between the stem cell group and the control group was statistically significant (P < 0.001), and the stem cell group was better than the control group.Twelve studies evaluated the effects of both groups in rabbits, including 214 rabbit discs. The main types of stem cells are BMSCs, ADSCs, and NPSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P < 0.001), and the stem cell group was better than the control group.Four studies evaluated the effects of both groups in mice, involving 80 mouse discs. Stem cell types are mainly ADSCs. The heterogeneity test showed significant heterogeneity between studies , so a fixed-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P < 0.001), and the stem cell group was better than the control group.Three studies evaluated the effects of both groups in sheep, involving 40 sheep discs. Stem cell types are mainly BMSCs. The heterogeneity test showed no significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P = 0.001), and the stem cell group was better than the control group.Three studies evaluated the effects of both groups in rats, including 123 rat discs. Stem cell types are mainly ADSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was not statistically significant (P = 0.355).Four studies evaluated the effects of both groups in sheep and included 70 sheep discs. The main types of stem cells are BMSCs and ADSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P < 0.001), and the stem cell group was better than the control group.Three studies evaluated the effects of both groups in rats, involving 60 rat discs. The main types of stem cells are BMSCs and ADSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P < 0.001), and the stem cell group was better than the control group.Four studies evaluated the effects of both groups in rats, involving 120 rat discs. Stem cell types are mainly WJ-MSCs and ADSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P < 0.001), and the stem cell group was better than the control group.Four studies evaluated the effects of both groups in mice, involving 80 mouse discs. Stem cell types are mainly ADSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P = 0.016), and the stem cell group was better than the control group.Two studies evaluated the effects of both groups in rabbits and included 24 rabbit discs. Stem cell types are mainly ADSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P < 0.001), and the stem cell group was better than the control group.A total of 5 studies compared the MRI signal score of the stem cell group and the control group. As shown in A total of 13 studies compare the T2 signal intensity of the stem cell group and the control group, as shown in P < 0.001, I2 = 92.6%), so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P < 0.001), and the stem cell group was better than the control group.Six studies evaluated the effects of both groups in rabbits, including 108 rabbit discs. Stem cell types are mainly BMSCs and NPSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P < 0.001), and the stem cell group was better than the control group.Four studies evaluated the effects of both groups in mice, involving 80 mouse discs. Stem cell types are mainly ADSCs. The heterogeneity test showed significant heterogeneity between studies , so a random-effects model was used. The comprehensive results showed that the difference between two groups was statistically significant (P = 0.003), and the stem cell group was better than the control group.Three studies evaluated the effects of both groups in rats, involving 100 rat discs. Stem cell types are mainly ADSCs. The heterogeneity test showed significant heterogeneity between studies (P < 0.001), disc height (P = 0.002), MRI index (P = 0.01), MRI signal score (P = 0.016), and T2 signal intensity (P < 0.001) had publication bias (P < 0.05), as shown in Figures 8We used Egger's method to detect publication bias. The test results showed that DHI , sheep (110 discs), rats (403 discs), and mice (240 discs). We used five imaging indicators, DHI, disc height, MRI index, MRI signal score, and T2 signal intensity, to evaluate the efficacy of stem cells in animal models. To our knowledge, this is the first study to systematically evaluate the efficacy of stem cells in animal models of disc degeneration over the past 20 years. In our study, the stem cell group covered both monotherapy and combination types, and subgroup analyses were performed to minimize heterogeneity and increase confidence in the results. Similar studies have not been done yet.The results of our study showed that the effect of the stem cell group in rabbits, mice, sheep, and rats was better than that of the control group in terms of DHI. In terms of disc height, the efficacy of the stem cell group in rats was better than that in the control group, while in sheep, there was no statistical difference in efficacy between the two groups. In terms of MRI index, the effect of the stem cell group in rats, mice, and rabbits was better than that in the control group. In terms of MRI signal score, the stem cell group was better than the control group in rabbits. In terms of T2 signal intensity, the stem cell group was more effective than the control group in rabbits, mice, and rats.Determining a suitable cell source is the premise and challenge for the successful establishment of disc regeneration therapy . Despiteβ1 (TGF-β1) [In the studies we included, ADSCs and BMSCs were the most commonly used stem cells, both of which are able to differentiate into chondrocytes , 39. BMSTGF-β1) , 41. The(TGF-β1) –45. Sinc , 41. Th(TGF-β1) .Although the above studies have confirmed that stem cell therapy can repair the intervertebral disc, it still has a limitation; that is, the content of type II collagen and proteoglycan in the repaired intervertebral disc is still lower than that of the normal intervertebral disc . TherefoFrom the perspective of the construction method of animal models, acupuncture caused annulus fibrosus damage, which was widely used in the included studies. This modeling method has the advantages of simple operation, low cost, and strong repeatability, and can cause the degeneration of the intervertebral disc in a relatively short period of time. However, the disadvantage is that there is a risk of infection, which may cause an immune inflammatory response . Also, aImaging evaluation is the most commonly used clinical evaluation method for disc degeneration. In terms of imaging indicators, MRI index, MRI signal score, T2 signal intensity, DHI, and disc height were the imaging indicators commonly used in the included studies . TherefoOur study has the following limitations. First, the heterogeneity of the studies is large. We believe that this may be related to the large differences in the injected dose of cells and the observation time between each study. Moreover, the imaging equipment used differs between studies, and the specific values of the images are calculated by third-party software, and the types of these softwares are also different, which may bias the results. Another point is the restriction of animal movement in the rearing environment, which was mentioned in only a few of the studies we included. We believe that differences in the amount of animal exercise caused by crowded and loose housing environments may also affect the degenerative process of the intervertebral disc, which is also a source of heterogeneity. Finally, we have yet to find studies examining the mechanical properties of the repaired disc, which may be the next step for refinement.In this study, through a large-scale meta-analysis of 34 studies, under multiple animal species and multiple evaluation indicators, the therapeutic effect of stem cells in the animal models of disc degeneration was observed from the perspective of imaging, which further demonstrated the role of stem cells in promoting intervertebral disc repair. However, it should be pointed out that it is not enough to demonstrate this repair effect from an imaging perspective alone. Researches on histology, biomechanics, and other aspects are also essential, which is also the direction of our next efforts.At present, most of the studies related to the repair of disc degeneration with stem cells are still in the stage of cell and animal experiments, and there is still a distance from clinical use. Encouragingly, there are already studies taking the first steps towards clinical application –52. In a


Beyond their specific experiment, video records of behavior have future value—for example, as inputs for new experiments or for yet unknown types of analysis of behavior—similar to tissue or blood sample banks in life sciences where clinically derived or otherwise well-described experimental samples are stored to be available for some unknown potential future purpose.Research using an animal model of obsessive-compulsive disorder employed a standardized paradigm where the behavior of rats in a large open field was video recorded for 55 minutes on each test. From 43 experiments, there are 19,976 such trials that amount to over 2 years of continuous recording. In addition to videos, there are 2 video-derived raw data objects: XY locomotion coordinates and plots of animal trajectory. To motivate future use, the 3 raw data objects are annotated with a general schema—one that abstracts the data records from their particular experiment while providing, at the same time, a detailed list of independent variables bearing on behavioral performance. The raw data objects are deposited as 43 datasets but constitute, functionally, a library containing 1 large dataset.Size and annotation schema give the library high reuse potential: in applications using machine learning techniques, statistical evaluation of subtle factors, simulation of new experiments, or as educational resource. Ultimately, the library can serve both as the seed and as the test bed to create a machine-searchable virtual library of linked open datasets for behavioral performance in defined conditions. If meaningful correlations are to be made between brain mechanisms and behavior, then the analysis of behavior will require as much sophistication and attention to detail as the analysis of the brain quinoline, agonist of dopamine D2/D3 receptors; QNP + IM: QNP coadministered with imipramine, a tricyclic antidepressant; QNP + clorgyline: QNP coadministered with clorgyline, a selective inhibitor of monoamine oxidase A; ROI: region of interest; SAL: 0.9% saline solution; U69593: N-methyl-2-phenyl-N-[-7-(pyrrolidin-1-yl)-1-oxaspiro[4.5]dec-8-yl]acetamide, agonist of opioid kappa receptors; VCR: videocassette recorder.The authors declare that they have no competing interests.Research using the animal model of OCD was supported by grants to Henry Szechtman from the Canadian Institutes of Health Research (CIHR) operating grants MOP-64424 and MT-12852, Ontario Mental Health Foundation (OMHF) Type A grant “Psychobiology of Security Motivation in an Animal Model of OCD,” and Natural Sciences and Engineering Research Council of Canada (NSERC) operating grant RGPIN A0544.These funding bodies had no role in study design, analysis, and interpretation of data and in writing the manuscript.H.S. was principal investigator on all studies, maintained the SPSS flat database for all studies, consolidated SPSS data records and data object files into a consistent framework, developed the annotation schema and annotated the datasets, wrote first draft of manuscript, and contributed to and approved final version of manuscript. A.D.G. was responsible for data processing and analysis in all studies, developed procedures and software scripts used in data processing and in analysis of compulsive checking, was the lead investigator and author on several publications from studies on compulsive checking, contributed to design of the FRDR collection and to the annotation schema, worked on writing the manuscript, and approved the final version. A.G.M. provided input on submission of the library to a public repository and contributed to design of the annotation schema and to the writing of the report.


Chronic mental diseases such as obsessive-compulsive disorder (OCD) are associated with a high disability rate. Some patients still do not improve their symptoms even with adequate cognitive-behavioral therapy and drug treatment. In the treatment of OCD, electroconvulsive therapy (ECT) is not considered a neuromodulation modality with sufficient evidence.This retrospective study aimed to determine the efficacy and associated risk factors of ECT in OCD patients.The study included 21 OCD patients who underwent ECT at a high-volume center in China between January 2009 and December 2020. The demographics and clinical characteristics of the patients were assessed using descriptive statistics. Based on Clinical Global Impressions–Improvement scale, patients were categorized into response and non-response groups. Clinical and demographic characteristics of two groups of patients were compared.2 = 6.252, P = 0.041).An analysis of 21 patients was conducted. In total, 12 patients (57.1%) responded to ECT, 11 patients (52.4%) reported side effects, and an average of 7 ECT sessions were administered. In terms of demographic, there was no statistically significant difference between the two groups. It is noteworthy that the non-response group reported more depression and schizophrenia related disorders comorbidities than the response group (χThe effectiveness of ECT in treating OCD is limited, especially in patients with refractory symptoms. Comorbidity with other mental disorders may affect the efficacy of ECT. It is a chronic and disabling mental illness called obsessive-compulsive disorder (OCD). Obsessions and compulsions are the most prominent symptoms. In the general population, lifetime prevalence is 2.3%, and 12-month prevalence is 1.2% , 5.Cognitive-behavioral therapy and serotonin reuptake inhibitors (SRIs) are recommended as first-line treatments for OCD by various treatment guidelines and expert consensus –8. It isElectroconvulsive therapy (ECT) is a technique used to treat mental illnesses by applying electrical current through the brain. There is ample clinical evidence that ECT is mainly used in major depressive disorder (MDD), bipolar disorder, schizophrenia –18. HoweAs a result, this retrospective study aimed to investigate ECT’s response rate among OCD patients and examine features associated with the response of OCD to ECT.This study included patients who received ECT therapy at West China Hospital of Sichuan University between January 2009 and December 2020. The OCD patients were included if they met the criteria outlined in the International Classification of Diseases, Tenth Revision (ICD-10) for the diagnosis. We analyzed only the first ECT series of patients who received more than one treatment series. Consequently, 21 OCD patients were included in the study.In accordance with the Declaration of Helsinki, it was approved by the Ethics Committee on Biomedical Research of West China Hospital of Sichuan University. Informed consent was not required for this study since it was exempt from institutional review.The ECT was performed with Thymatron System IV . As part of our study, we collected data on ECT treatments, including the number of treatments and electrode placement. Whether or not ECT was required and the ECT protocol was determined by the clinical psychiatrist according to the patient’s condition. The diagnosis, treatment options, and risks and benefits of ECT were explained to patients and their legal guardians and informed consent was obtained prior to the start of ECT.Typically, ECT administration was performed 3 times per week . The electrodes were positioned bilaterally in the temporal region. Under the supervision of an anesthesiologist and psychiatrist, propofol and succinylcholine were given separately for anesthetic and muscle relaxant.Demographic information was collected, including age, gender, and education level. In this study, we collected clinical factors, such as the age at which OCD onset occurred, the course of illness, comorbidities, and treatment histories. Furthermore, we examined ECT-related characteristics, such as the number of ECT sessions and the reasons for administering ECT.We evaluated the charts independently by two authors (KL and JL) to determine their response to ECT. There was a consensus reached by reviewing disagreements with the experts (WD). Improvement of OCD were assessed using the Clinical Global Impressions-Improvement scale (CGI-I). On the CGI-I, a response was defined as a score of 1 (very much improved) or 2 (much improved). Patients or psychiatrists will record all side effects reported spontaneously or observed. Records and CGI-side effects scales were used to assess side effects.t-tests or Mann–Whitney U test for continuous variables. For categorical variables, we used the Chi-square test or Fisher’s exact test. The significance of the results was defined as P values < 0.05.All statistical analyses were conducted using SPSS . We performed descriptive statistical analyses on all variables, including demographic characteristics, clinical factors, and ECT-related characteristics. Descriptive statistics involves computing means, standard deviations, and frequencies for continuous and categorical variables. Comparisons between groups were made with independent samples Twenty-one OCD patients receiving ECT were included in the study. p value for comparison between the two groups was 0.041 (χ2 = 6.252). In addition, we separately compared the effects of comorbid MDD group with comorbid schizophrenia related disorders group and found no statistically significant difference between the two groups . The comorbidity rate was higher in the non-response group (77.8%) than in the response group (16.7%).As shown in To our knowledge, this is the first cohort of Chinese OCD patients treated with ECT. In this study, the effectiveness of ECT treatment of OCD and the occurrence of side effects were examined and predictors of efficacy were identified. In the total sample, at least 57.1% of patients reported positive responses to ECT. The OCD is frequently comorbid with MDD or schizophrenia-related disorders, with a 33.3 and 9.5% comorbidity rate, respectively. The effectiveness of ECT may be reduced if it is comorbid with other mental disorders, which is notable.While this study suggests ECT has 57.1% response rate for OCD, patients are highly heterogeneous. The results of this study were similar to previous observational studies in which ECT significantly improved symptoms of OCD in most patients . It mighIt is interesting to note that 13 (61.9%) patients who received ECT had refractory or severe symptoms, but other patients required ECT for comorbidities of other severe psychiatric symptoms that needed to be addressed. It is noteworthy that patients with OCD who have comorbid MDD or schizophrenia-related disorders were less effective than those without. The results of this study are consistent with previous studies . The obsMoreover, ECT has been found to be effective in treating both obsessive-compulsive symptoms and depression symptoms , 49, 50.It is also important to note that there are some limitations to this research. Due to the retrospective nature of this study, we were unable to control for various confounding variables. The future will demand large-scale, high-quality randomized controlled studies to fulfill clinical practice requirements. Secondly, statistical power was limited by the small sample, which may explain some of the non-significant findings. Moreover, we lack quantifiable metrics for comorbid symptoms, such as catatonic symptoms –53. As tAs a result of this study, we can conclude that ECT for OCD has limited efficacy, especially in refractory patients. There is a possibility that comorbid MDD or schizophrenia related disorders could adversely affect ECT. Therefore, this study needs to be further verified by a randomized controlled trial involving a large sample size.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Ethics Committee on Biomedical Research of West China Hospital of Sichuan University. Written informed consent for participation was not required for this study in accordance with the national legislation and the institutional requirements.KL: conceptualization, formal analysis, data curation, and writing – original draft. JL: formal analysis and data curation. BC: data curation, investigation, and methodology. JW: data curation, project administration, supervision, and writing – review and editing. WD: conceptualization, data curation, project administration, supervision, and writing – review and editing. All authors contributed to the article and approved the submitted version.