We describe the multimodal neuroimaging dataset VEPCON (OpenNeuro Dataset ds003505). It includes raw data and derivatives of high-density EEG, structural MRI, diffusion weighted images (DWI) and single-trial behavior . Visual evoked potentials (VEPs) were recorded while participants (n = 20) discriminated briefly presented faces from scrambled faces, or coherently moving stimuli from incoherent ones. EEG and MRI were recorded separately from the same participants. The dataset contains raw EEG and behavioral data, pre-processed EEG of single trials in each condition, structural MRIs, individual brain parcellations at 5 spatial resolutions (83 to 1015 regions), and the corresponding structural connectomes computed from fiber count, fiber density, average fractional anisotropy and mean diffusivity maps. For source imaging, VEPCON provides EEG inverse solutions based on individual anatomy, with Python and Matlab scripts to derive activity time-series in each brain region, for each parcellation level. The BIDS-compatible dataset can contribute to multimodal methods development, studying structure-function relations, and to unimodal optimization of source imaging and graph analyses, among many other possibilities. Machine-accessible metadata file describing the reported data: 10.6084/m9.figshare.17074685 EEG potentials registered at scalp electrodes result from synchronous activity in large populations of neurons that are distributed across cortical and subcortical areas4. Visual stimulation gives rise to a fast sequence of well-known EEG components that reflect initial processing at latencies before ~100 ms6, subsequent object and recurrent processes9, and later components that reflect target detection, integration and decisions12. The VEP provides a millisecond by millisecond recording of whole-brain activity dynamics, and has a rich distribution of temporal frequencies that provides further insight into the functionality of brain processes14.Visual evoked potentials (VEPs) have a long record of shedding light on the spatial and temporal dynamics of large-scale neural processing in the brain18. The anatomical constraints determine what electrical fields from a neural activity source would look like at the recording electrodes on the scalp, given the conductivities of various tissues and fluids lie in between. An inverse solution translates the recorded electrical potential field back to a pattern of distributed source activity in the brain. Source localizations from EEG are necessarily coarse, as compared to fMRI, and improving them further is an active field of research and helps understand the activity dynamics within areas, and their inter-relatedness22.From VEPs recorded across the scalp, the underlying distributed patterns of brain activity can be estimated using an inverse solution based on anatomical constraints26. This structural connectivity can be inferred with diffusion weighted imaging (DWI)28. The resulting connectome constitutes a road map of sorts over which activity can propagate between areas, and in an important sense constrains how activity within an area can evolve through the influences it receives from others30.Activity in each area can influence activity throughout the brain in a few steps, due to the dense connectivity of cortico-cortical and cortico-subcortical fibers31 combines raw EEG data, T1-weighted (T1w) MRI, and DWI for 20 human participants, with as derivatives, VEPs, inverse solution matrices, brain parcellations and connectomes at 5 different spatial scales 37.The VEPCON dataset31, and raw data are structured following the MRI and EEG Brain Imaging Data Structure standards (BIDS39). We expect the data to be useful for the development and benchmarking of multimodal analysis methods that combine functional and structural information, for exploring structure-function relations, and for controlling whether and how the level of parcellation affects results. We also expect unimodal reuse value for development and benchmarking of EEG source-imaging approaches and functional connectivity analyses. Beyond this, the availability within the same participant of single-trial behavior, EEG data, inverse solutions, anatomical and diffusion MRI, and connectomes based on four different indices allows for many types of analyses that can generate new hypotheses about dynamic brain function in human visual processing and behavior.The dataset is publicly available on OpenNeuro40. Nineteen participants were right-handed, one was left-handed. All participants provided written informed consent before the experiment. The experimental procedures complied with the Declaration of Helsinki and were approved by the regional ethics board .Twenty participants  took part, recruited from the local student population . Participants had normal or corrected-to-normal vision. Before the experiment visual acuity was tested with the Freiburg Acuity test, and a value of 1 had to be reached with both eyes openEEG data were recorded while participants performed a face detection and a motion discrimination task see Fig. . The ord41. In the face detection task, each trial lasted 1.2 s and started with a blank screen (500 ms). After the blank screen, one image (either a face or a scramble image of a face) was presented at the center of the screen for 200 ms and participants had the remaining 1000 ms to respond. The task was to report whether they saw a face or not (yes/no task) by pressing one of two buttons in a response box with their right hand. After the response and a random interval (from 600 to 900 ms), a new trial began. The experiment consisted of four blocks of 150 trials each, for a total of 600 trials, i.e., 300 with faces and 300 with scrambled faces. Faces and scrambled faces were randomly interleaved across trials.Face stimuli were female and male faces  taken from online repositories and cropped with a Gaussian kernel to smooth the borders. Scrambled images were obtained by fully randomizing the phase spectra of the original imagesIn the motion discrimination task, motion stimuli were dot kinematograms presented on a circular frame at the center of the screen . For coherent motion, 80% of the dots were moving toward either the left or right, with the remaining 20% moving in random directions. For incoherent motion stimuli, all dots moved randomly. Each trial started with a blank interval of 500 ms followed by a centrally presented stimulus for 300 ms . Responses were collected using a ResponsePixx response box (VPixx technologies).Stimuli were generated using PsychopyEEG data were recorded at a sampling rate of 2048 Hz with a 128-channel Biosemi Active Two EEG system  in a dimly lit and electrically shielded room. Signal quality was ensured by monitoring and maintaining the offset between the active electrodes and the Common Mode Sense - Driven Right Leg (CMS-DRL) feedback loop under a standard value of ± 20 mV. After each recording session, individual 3D electrode positions were digitized using an ultrasound motion capture system .44. EEG data were downsampled to 250 Hz and detrended . Line noise (50 Hz) was removed via spectrum interpolation45. Data were then segmented into epochs and time locked from −1500 to 1000 ms from the stimulus onset in both tasks. Data from participant 05 (Face and Motion tasks) and 15 (Motion task) was not further processed and discarded due to excessive noise. Bad channels and epochs were identified and removed before preprocessing. Remaining physiological artifacts were isolated using an independent component analysis (ICA) decomposition (FastIca). Bad components were labelled by crossing the results of a machine-learning algorithm  with the criterion of >90% of total variance explained and removed manually. Bad channels were then interpolated using the nearest-neighbor spline method and data were re-referenced to the average reference.Offline, data were preprocessed using EEGLAB 14.1.146 and custom-made scripts (see Matlab and Python examples in the code/ directory). Source reconstruction was based on Cartool-segmented individual MRI T1w data, co-registered individual electrode positions, and the LORETA47 and LAURA15 algorithms . Leadfields were calculated for each of the around 5000 freely oriented dipoles while limiting the solution space to voxels within the gray matter mask46 provided by CMP3.EEG source imaging was performed using Cartool (v3.80)2). DTI data were acquired with a spin echo single shot EPI pulse sequence and a diffusion sensitizing gradient set of 30 different directions and 5 diffusion-free B0 scans .MR data from the same 20 subjects was acquired on a General Electrics Discovery MR750 3 T MRI clinical scanner at the cantonal hospital in Fribourg, Switzerland, using a 32-channel head coil. The acquisition included anatomical T1-weighted images and DTI. T1-weighted images were acquired as rapid-gradient echo (MPRAGE) volumes using a COR FSPGR BRAVO pulse sequence  2.2.0. Finally, connectivity matrices at 5 different spatial scales were built by considering all fiber streamlines connecting parcels according to the following connectivity measures: number of fibers, fiber density, average and median Fractional Anisotropy (FA), and Mean Diffusivity (MD).For each participant, structural connectivity matrices were estimated from the reconstructed fiber orientation distribution (FOD) image using the SD_stream deterministic streamline tractography algorithm implemented in MRtrix 3.0.0-RC131, and is fully BIDS compatible (v1.4.1). Below, we describe all data records following the directory structure.The VEPCON dataset is available via the Open Neuro repositoryThe main directory contains descriptor files detailing the dataset, and the age and sex of each participant. Subfolders (sub- <label>/) hold the raw EEG data (eeg/) in .bdf format, as well as the MRI T1w images (anat/) and diffusion weighted images (DWI) (dwi/) in the compressed Nifti format, with acquisition parameters stored in their associated sidecar .JSON file. With respect to raw EEG data, description of timing and other event properties are stored in *_task- <label>_events.tsv files and the corresponding column descriptor (*_task- <label> _events.json), for each task (task-faces and task-motion). The events file contains a table listing for each single trial the stimulus onset relative to recording onset (sec), condition specifiers, response made, response evaluation, reaction time and whether the trial was discarded or not in preprocessing. Trials with behavioral errors or reaction times slower than 200 ms were marked as outliers. For each electrode the digitized electrode positions  are listed in the *_electrodes.tsv file, and status information (good/bad) is listed in the *_channels.tsv file.The derivatives/ directory includes a dataset_description.json file for each derivatives dataset  at the root level. Any subject-specific datatype-specific derivatives are housed within each subject/datatype’s directory (sub- <label>/<datatype>) where <datatype> can be “anat”, “dwi”, “eeg”, “xfm” or “connectivity”. The output reports generated by MRIQC, and CMP3 derivatives of anatomical and diffusion MRI are in the mriqc- <version> cmp- <version>) folders, where the version of each software used is encoded by the <version> label . EEG derivatives obtained with EEGLAB (eeglab- <version>) and Cartool (cartool- <version>) are contained in their respective folders and organized to comply with BIDS to the fullest possible extent (specifications for some of these files are currently under development).In the eeglab/ directory, each eeg/ participant’s folder contains the preprocessed single trial epochs . The task-faces files contain epochs for face stimuli and control stimuli, the task-motion files contain epochs for coherent and incoherent motion stimuli. The directory holds summaries of preprocessing and data cleaning for the motion and face task in two separate .tsv files. These files list for each participant the proportion of channels, epochs and ICA components removed during preprocessing.47 are in the respective .is files; they map observed patterns of EEG potentials to a distribution of 3D dipoles across source points. The rois/ folder lists files that indicate which source points belong to what area , according to each parcellation level, with a Cartool readable (.rois) and a Python readable version (.pickle.rois).In the cartool/ directory, each eeg/ participant’s folder holds a text file (.xyz) with individual electrode positions in mm, co-registered to the participant’s head . The .spi file is a similar text file listing x, y, z coordinates and a text label for each sourcepoint for which an inverse solution was calculated. The inverse solution matrices for LAURA and LORETAThe cmp/ directory contains anatomical, diffusion and structural connectivity data for each participant. All T1w-derived data are placed in the anat/ folder, which includes the brain segmentations and parcellations in the native T1w space and in the native DTI space (_space-DWI_). All DTI-derived data are placed in the dwi/ folder. It includes the preprocessed DTI, the FOD image and the final tractogram used to build the connectivity matrices. The connectivity/ folder holds connectomes at each parcellation level, with a Matlab readable (.mat) and Python readable (.gpickle) version. Each connectome file contains the following metrics: number_of_fibers, fiber_density, mean_FA and median_FA, mean_MD and median_MD. The transformations from the native T1w space to the native DTI space applied to the parcellations are stored in the xfm/ folder. The generated .pdf in the group/ folder visualizes each individual’s connectome using the number of fibers metric, for parcellation scale 1.53. Code to run Connectome Mapper 3 on all participants that can be found in the ConnectomeMapper-Docker/ folder. The code to deface anatomical MRIs52 is provided in the Deface-Python/ folder, together with the generated log file and a report in pdf format. In Source-Reconstruction-Python/, the main.py script performs all steps for source reconstruction, with comments that explain each step. The utils.py contains all necessary functions to run main.py. The Source-Reconstruction-Matlab/ folder provides the equivalent in Matlab code, with the ESI_RECONSTRUCTION.m script performing all steps of source reconstruction, and dependencies and utilities as separate files.The code/ directory provides code for preprocessing and derivation steps for MRI and DWI data, as well as sample scripts to derive time-series of activity per ROI from the preprocessed EEG data in Python and Matlab. Each code folder has its own README file with afor more specific user guide and documentation to get started. The MRIQC-Docker/ folder holds the code to run MRIQC on all participants. All outputs generated by MRIQC to support quality assessment of T1w data can be found in the same directoryAnalysis of proportion correct and reaction times showed that participants behaved according to task instruction. In the face detection task, the average accuracy was 97 ± 2% of correct responses, with mean reaction times of 501 ± 60 ms. Trials with behavioral errors or reaction times slower than 200 ms were marked as outliers (mean proportion of outliers 0.03 ± 0.02).In the motion discrimination task, the average accuracy was 90 ± 14% and mean reaction times were 680 ± 90 ms. The mean proportion of outliers was 0.10 ± 0.14. One participant (number 16) inverted the response keys for several trials in the motion task, leading to an outlier accuracy value (36% of correct responses).EEG data were visually inspected and noisy trials and channels were excluded from further analysis (see EEG recording and preprocessing). Remaining ocular, muscle and other artifacts were removed using ICA. For the face task, the average proportion of channels removed across participants was 0.12 ± 0.07, range 0.02–0.28; mean proportion of epochs removed due to non-stereotyped artifacts, peristimulus eye blinks and eye movements: 0.04 ± 0.06, range 0.002–0.26); proportion of ICA components removed: 0.05 ± 0.03, range 0.01–0.15. For the motion task, the average proportion of channels removed was 0.14 ± 0.06, range 0.02–0.23, the average proportion of epochs removed was 0.03 ± 0.03, range 0.005–0.13); proportion of ICA components removed: 0.05 ± 0.04, range 0.008–0.13.Figure 53. Results were summarized in reports at both individual and group levels, which are included in the mriqc/ derivatives folder.After recording MRI data were visually inspected and checked for neurological anomalies by the radiologist. Quality of T1w data was further inspected quantitatively by running MRIQC v0.16.1, an automated and robust quality control tool for T1w data which derives a set of 56 different image quality metrics to characterize image quality at different levels such as noise, motion or imaging artifactsDe-identification of T1w data was checked individually and supported by the creation of a PDF report that is available in the code/ folder.For each participant, the quality of the output after each processing step of the CMP3 pipelines was assessed using its graphical user interface, where we inspected individually the different Freesurfer outputs, brain parcellations, DTI data after preprocessing, co-registered T1w-derived parcellations with DTI data, the estimated FOD image, the reconstructed tractogram, and the reconstructed connectomes. Visual inspection did not reveal any artifacts.38 and EEG39.For optimal sharing and re-usability, raw data of the dataset conforms to BIDS standards for MRI54, NeuroPycon55, Fieldtrip56 and SPM57. The code also shows how to calculate one time-series of activity from all source points contained in a region using singular-value decomposition. Source activity for around 5000 freely oriented dipoles was extracted from all the source points inside each cortical area, as defined by the parcellation, and projected to a representative single direction using singular-value decomposition32.Python and Matlab example code in Source-Reconstruction-Python/ and Source-Reconstruction-Matlab/ shows how to import the various raw and derivative files. This assures compatibility with commonly used EEG and MRI software, including MNE-Python58.For creating new inverse solutions, new forward models can be created from the T1w images, using the individual electrode coordinate files. When new leadfields are generated using the defaced MRIs, small differences can occur, but are unlikely to pose problems for the intended reuse purposes


Plant parasitic nematodes (PPNs) are responsible for causing many plant diseases and are extremely difficult to control at present. Currently, due to the negative effects of chemical agents on the environment and human health, the development of new biological pesticides has become an important part of plant nematode control. Nematophagous fungi refers to a class of fungi that kill plant nematodes. Notably, a large number of nematophagous fungi resources remain to be studied. The objective of our study was to use in vitro screening to identify nematophagous fungi and select strains that were highly active against nematodes, providing a primary research for the development and utilization of new nematophagous fungi.Globodera spp. and Heterodera spp., identified as Volutella citrinella. The hyphae of V. citrinella produced ring structures of variable size and exhibited predatory and nematicidal activity. The hyphal predation rates (in vitro) against three species of nematodes, Aphelenchoides besseyi, Bursaphelenchus xylophilus, and Ditylenchus destructor, averaged 59.45, 33.35, and 50.95%, respectively, while the fermentation broth produced by the fungus exhibited mortality rates of 100, 100, and 55.63%, respectively, after 72 h.A new nematophagous fungal strain (GUCC2219) was isolated from cysts of possibly V. citrinella is a new strain with nematophagous properties, which are a novel discovery. At the same time, this is the first report of nematicidal and nematode predation activity in the genus Volutella.The online version contains supplementary material available at 10.1186/s12866-021-02385-x. Plant parasitic nematodes (PPNs) are major pathogens of many crops and comprise more than 4100 species globally . A varieGlobodera and Heterodera spp.), root knot nematodes (Meloidogyne spp.), foliar nematodes (Aphelenchoides spp.), pine wilt nematode (Bursaphelenchus xylophilus), Ditylenchus destructor, Anguina tritici [A. besseyi, B. xylophilus and D. destructor are especially important in China. Aphelenchoides besseyi mainly damages rice, whose leaves exhibit white tips after becoming infected and then become necrotic. The flagged leaves are curled and deformed, inflorescences are reduced in size and the heads do not develop mature grains, resulting in a decrease in yield [A. besseyi has been listed as one of the ten most harmful plant nematodes due to its potential impact on yield [Bursaphelenchus xylophilus is responsible for causing many tree diseases globally [Monochamus alternatus feeding on healthy pine trees [Ditylenchus destructor is also one of the most destructive plant pathogenic nematodes [Species of plant nematodes that impact agriculture include cyst nematodes  on Japanese pachysandra  [V. buxi causing Boxwood Volutella stem blight or canker on boxwood (Buxus spp.) [Volutella, which are worthy of further study.Approximately 380 species of nematode-trapping fungi have been reported from different regions of the world . Howeverhe world . The gen species . Accordiciculosa ) oxidizeciculosa . In addiciculosa  reportedminalis) ; V. buxius spp.) . There wGlobodera spp. and Heterodera spp. of potatoes growing in Weining County, Guizhou Province, China. A total of 147 isolates were obtained, mainly fungi. Their nematicidal properties were evaluated by in vitro screening. One isolate (GUCC2219) was found to exhibit high hyphal predation rates in vitro against Aphelenchoides besseyi, Bursaphelenchus xylophilus, and Ditylenchus destructor, with predation rates averaging 59.45, 33.35, and 50.95%, respectively, while its fermentation broth exhibited mortality rates of 100, 100, and 55.63%, respectively, after 72 h. The GUCC2219 fungal isolate was identified as Volutella citrinella based on morphological observations and phylogenetic analysis of the genetic sequence of the internal transcribed spacer (ITS) and large subunit (LSU) regions of DNA. This is the first report of nematicidal and nematode predation activity in the genus Volutella.In the present research, colonizing microorganisms were isolated from cysts of possibly Volutella citrinella (DAOM: 226720) and exhibited high sequence similarity, 99% (530/531) and 100% (877/877) to its ITS and LSU gene regions, respectively.The sequences of the polymerase chain reaction (PCR) products obtained from strain GUCC2219 were uploaded to GenBank and subjected to Basic Local Alignment Search tool (BLAST) analysis. The highest similarity was obtained to accession numbers MZ148447 (ITS) and MZ148449 (LSU). In the phylogenetic tree  and cornmeal agar (CMA) for 14 days at 25 °C. Observations indicated that colonies were superficial on PDA, reaching 20-30 mm diam. A purple-red pigment was produced on the underside of the colony  at 1% water agar (WA) revealed that it produced a large number of hyphal rings through the twining of mycelia  due to their increased entanglement with hyphae after 24-48 h of coincubation. Two days later, the hyphae had started to attach to the surface of entrapped nematodes and completely engulf them. After 72 h of coincubation, the number of fungal mycelia composing the rings increased, the body wall of the entrapped nematodes dissolved, the internal contents of the nematodes consumed, and only traces of the entrapped nematodes remained  exhibited significant predatory activity against the three species of nematodes at different points in time. In general, the percentage of trapped nematodes increased with the time of coincubation for all three species of nematodes (Table V. citrinella (GUCC2219) was greatest against A. besseyi, with an average percentage of 59.45% at 72 h, followed by D. destructor and B. xylophilus, with averages of 50.95 and 33.35%, respectively (P < 0.05).Under in vitro conditions, the GUCC2219 strain of V. citrinella (GUCC2219) fermentation filtrate was significantly different from that of the potato dextrose broth (PDB) medium alone  was the highest against A. besseyi, producing mortality levels of 21, 27, 100, and 100% at 12, 24, 48, and 72 h, respectively  exhibited different levels of nematicidal activity against the three different species of nematodes.The nematicidal activity of the undiluted fermentation filtrate of ely Fig. A. The moA. besseyi (68%), B. xylophilus (48%) and D. destructor (30%) at 72 h (P < 0.05).  and D. destructor (15%) (P < 0.05)  were mainly produced under low-nutrient WA, which mainly form three ring types: small, regularly shaped; large, regularly shaped and irregularly shaped rings of various sizes. There were some differences from currently reported fungal nematode traps, such as adhesive nets, adhesive knobs, adhesive branches, and constricting rings [V. citronella, are entangled by the hyphae and rings. Of interest is the fact that the rings turn a golden color when the nematodes are caught. This feature has not been reported for other fungi that uses the same method of entrapment. We speculate that a chemical substance that are released when a nematode is trapped, might be responsible for the color change.Nematode traps are generally produced by fungi under low-nutrient or nutrient-deficient conditions but can also be induced by other agents, such as abscisic acid, amino acids, bacteria, nematodes, etc. . The trang rings . NematodBeauveria bassiana (Snef2621) against A. besseyi, exhibiting a mortality rate of 64.85% at 48 h [V. citrinella (GUCC2219) fermentation filtrates have higher nematicidal activity, and the mortality rate reached 100% at 48 h. Wu et al. [Myrothecium verrucaria, whose fermentation filtrates exhibited a high mortality rate (96.1%) against B. xylophilus. In our research, V. citrinella (GUCC2219) fermentation filtrates also had high activity against B. xylophilus, with a mortality rate of 100%. The study by Zhang et al. [Bacillus subtilis 1 × 109 spore/g wettable powder (WP) and Trichoderma fertilizer ≥2 × 108 spore/g dustable powder (DP) had nematicidal activity against D. destructor, the corrected mortality rate reached 73.45 and 44.76% at 72 h, respectively. The activity of V. citrinella (GUCC2219) fermentation filtrates against D. destructor (51%) was between those of these two microbial agents. The fermentation filtrate of V. citrinella (GUCC2219) had nematicidal activity against all three species of nematodes that were also evaluated, although the level of nematicidal activity differed among the three different species. Collectively, our data provide information on a new fungal strain (V. citrinella) with nematicidal activity.Previous studies have reported nematicidal activity of the fermentation filtrate of some fungi, such as  at 48 h . In contu et al.  reportedg et al.  showed tPaecilomyces lilacinus and Verticillium chlamydosporium [Volutella.Compared with the previously reported commercial preparations of fungi, such as osporium , they arosporium , and basA. besseyi, B. xylophilus, and D. destructor, and the mycelia also changed from pale white to light yellow after capturing a nematode. The GUCC2219 strain was identified as V. citrinella based on morphological observations and a phylogenetic analysis of the DNA sequences of its ITS and LSU regions. Our finding represents a new record in China. The fungus was able to produce hyphae that formed complex ring structures capable of trapping the three different species of nematodes that were evaluated. Our results clearly demonstrated the predatory properties of the hyphal rings formed by V. citrinella (GUCC2219), representing a new and novel discovery. V. citrinella (GUCC2219) produced a constant number of hyphal ring structures in a low-nutrient environment and the fermentation filtrate against the three species of nematodes in a short time without being concentrated, which presumably contributed to its strong predatory activity and nematicidal properties. Our research has laid the foundation for the application of the nematophagous fungi (GUCC2219). According to the results of biological characteristics and nematicidal ways, the biological pesticides can be further developed. One is the spore suspension, and the other is nematicidal fermentation filtrate. For seed-borne diseases such as A. besseyi, the seeds can be soaked before sowing, or the inoculant can be sprayed on the leaves and stems of the rice; B. xylophilus, which are parasitic nematodes in pine trees, spores or nematicidal fermentation filtrate can be injected into the plants through a syringe; D. destructor, which parasitizes mainly tubers, bulbs and root crops, root irrigation can be carried out to achieve the purpose of preventing nematode diseases. Of course, these are only a bold speculation, and further research is needed to prove the applicability of this strain in the field.In the present study, we report a new nematophagous fungal strain (GUCC2219) that was isolated from cysts formed by potato cyst nematodes. The GUCC2219 strain exhibited the potential to control three species of nematodes in vitro, Soil samples were collected from 18 fields of potato plants growing in Weining County, Guizhou, China, that were infected by PCNs. In each field, ten plots of a 5 × 5 m grid were selected surrounding infected potato plants, and in each grid, an approximate volume of 250 ml of soil was collected from rhizosphere zone (0-20-cm depth). The individual samples of each plot were collected and mixed in a bucket to obtain a single composite sample . Each co2O2 for 3 min following three washes with distilled water. The surface-sterilized cysts were individually placed onto 1% WA plates. Plates were incubated at room temperature and monitored regularly. Fungal mycelia growing from the cultured cysts were then recultured several times on new PDA plates. The obtained isolates were initially screened for nematode predation activity. A total of 44 isolates were screened (A. besseyi), one isolate exhibited nematode predation activity  were collected from rice growing in Dushan County, Guizhou Province, China, and D. destructor was provided by the Nematode Laboratory, Fujian Agricultural and Forestry University. The nematodes were propagated on carrot callus [Samples of t callus . The nemBursaphelenchus xylophilus was provided by the Center for Research and Development of Fine Chemicals, Guizhou University, Guiyang, China. It was cultured on PDA plates containing Botrytis cinerea [B. cinerea, and the plates were incubated at 28 °C until the B. cinerea colony was completely consumed. The nematodes were separated from B. cinerea cultured using the Baermann funnel method [ cinerea , which wl method . A nematFor the preparation of PDB medium, 200 g potatoes were added to 1 L of distilled water and boiled for 30 mins. The mixture was then filtered through double gauze, and 20 g dextrose was added to the filtrate. Subsequently, 100 ml aliquots of the prepared PDB medium were placed into 250 ml conical flasks and sterilized in an autoclave at 121 °C for 30 min. A single pure culture of the GUCC2219 isolate was cut into small pieces approximately 5 mm in diameter using a sterilized cutting blade. Five pieces were added into to 100 ml of sterilized PDB medium, which was then placed on a rotary shaker at 28 °C set at 200 rpm for 7 days. Afterwards. the medium was filtered and stored at 4 °C.The GUCC2219 isolate was inoculated on two types of different media, PDA and CMA, and cultured at 25 °C for 15 d. Photographs of the colonies, mycelia, and mycelial structures were taken using a stereomicroscope . Sections of the fungal colonies were made with the assistance of a stereomicroscope (Leica S9i) and mounted in water. Photomicrographs of conidiophores and conidia were taken using a compound light microscope (Zeiss Scope 5) equipped with an AxioCam 208 color camera.2O, 1 μl forward and reverse primers (10 μM/μl), and 1 μl DNA template. PCR products were commercially sequenced by using the same PCR primers used in the amplification reactions by SinoGenoMax (Beijing).The GUCC2219 fungal isolate was grown on PDA at 25 °C for 15 d. The resulting mycelia were then scraped off the surface of the plate with a sterile scalpel. Total genomic fungal DNA was extracted using a BIOMIGA Fungus Genomic DNA Extraction Kit  following the manufacturer’s protocol. ITS and LSU were PCR-amplified using the primer pair ITS5/ITS4  and LR0RSequences of each gene generated from forward and reverse primers were assembled with BioEdit v.7.2.5 , and conIn the preliminary screening, a 5 mm-diameter disc of mycelium was taken from the margin of the fungal isolates, transferred to the center of a Petri dish containing 1% WA, and then incubated in the dark at 25 °C for 2 weeks. After the incubation period, Petri dishes were inoculated with a 1 ml nematode suspension containing 1000 nematodes. The nematode suspension was divided into 4-5 drops and spread uniformly inside the periphery of the fungal colonies. Plates without fungi served as controls. The morphology and the average size of the hyphal rings were observed and recorded. Four replicates of each isolate were utilized. The number of nematodes captured in the hyphal rings was counted using a stereomicroscope . Microscopic images were captured with a compound light microscope (Zeiss Scope 5) equipped with an AxioCam 208 color camera.A 100 μl suspension of nematodes containing approximately 100 nematodes was placed into wells of a 96-well culture plate containing different concentrations  of the fermentation broth. Control wells contained distilled water. The plate was incubated at 28 °C for 72 h. Nematodes were microscopically monitored after 12, 24, 48, and 72 h. At each timepoint, the nematodes were washed and transferred into distilled water to observe their motility as an indication of nematicidal activity. Nematodes were considered dead when they remained immotile upon probing with a fine hair needle, and percentage mortality was calculated. Four replicates were analyzed for each concentration and for the control.P < 0.05 using Duncan’s multiple range. All statistical analyses were conducted in MS Excel and SPSS statistics (version 19.0) software. Figures were generated using Origin 2018.The data were subjected to two-way analysis of variance (ANOVA), with concentration and posttreatment time (exposure period) serving as the main treatment effects and concentration x time as the interaction. Significant differences between means were determined at Additional file 1: Schedule 1. Some genera or species distributed by nematophagous fungi.


Growth differentiation factor 15 (GDF15) has been associated with food intake and weight regulation in response to metabolic stress. In animal models, it has been noted that it may play a role in the progression of non-alcoholic fatty liver disease (NAFLD), the leading cause of chronic liver disease in children.In the current study, we explored the association of circulating plasma concentrations of GDF15 with NAFLD in youth with overweight/obesity, and whether changes in plasma concentrations in GDF15 parallel the changes in intrahepatic fat content (HFF%) over time.n = 11) or regressed (n = 11) in HFF% by more than 30% of original over a 2-year period.Plasma GDF15 concentrations were measured by ELISA in 175 youth with overweight/obesity who underwent an oral glucose tolerance test (OGTT) and magnetic resonance imaging (MRI) to assess intrahepatic, visceral, and subcutaneous fat. Baseline fasting GDF15 concentrations were measured in twenty-two overweight/obese youth who progressed (z-score (BMIz), and visceral fat (P = 0.002). During the OGTT, there was a decline in plasma GDF15 concentrations from 0 to 60 min, but GDF15 concentrations returned to basal levels by the end of the study. There was a statistically significant association between change in HFF% and change in GDF15  over ~2 years of follow-up.Youth with NAFLD had significantly higher plasma concentrations of GDF15 than those without NAFLD, independent of age, sex, ethnicity, BMI These data suggest that plasma GDF15 concentrations change with change in intrahepatic fat content in youth with overweight/obesity and may serve as a biomarker for NAFLD in children. Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in pediatrics and affects ~30% of children and adolescents with obesity in the United States –4. DespiRecent studies have shown that GDF15 is associated with NAFLD in adults. In particular, GDF15 represents an independent determinant of fibrosis severity in NAFLD: (1) patients with more severe chronic liver diseases have proportionately higher GDF15 values and (2) plasma GDF15 concentrations are associated with liver fibrosis regardless of diabetes status , 13.So far, there are no studies investigating whether plasma GDF15 concentrations in youth with obesity are associated with NAFLD. In this study, we sought to determine whether GDF15 plasma concentrations are associated with (1) intrahepatic fat content, (2) changes in intrahepatic fat accumulation over time, and (3) the metabolic phenotype related to NAFLD in youth with obesity. To achieve our aims, we measured GDF15 in 175 youth with overweight/obesity who underwent an oral glucose tolerance test and an MRI to measure intrahepatic fat content. Moreover, we analyzed changes in GDF15 over time in 22 youth who were followed-up for ~2 years and also showed changes (increase or decrease) in intrahepatic fat content of at least 30%. Herein, we show that in youth with overweight/obesity, GDF15 plasma concentration is associated with NAFLD and NAFLD-related phenotypes and that changes in plasma GDF15 concentration are driven by changes in intrahepatic fat content.th percentile according to U.S. Centers for Disease Control and Prevention (CDC) growth charts [A cross-sectional cohort of children and adolescents with overweight/obesity was identified from the Yale Pediatric Obesity Clinic main research study cohort . A multiethnic group of 175 youth with overweight/obesity between the ages of 8 and 21 years was selected based on protocol inclusion and exclusion criteria. Per the study protocol, participants had a BMI ≥ 85h charts , and wern = 10) and without (n = 10) NAFLD, matched for age, sex, BMIz, and ethnicity, during an OGTT. Longitudinal analyses were performed in a subgroup of 22 youth with overweight/obesity who were followed up for ~2 years (1.79 years ± 0.178) and either increased (n = 11) or decreased (n = 11) HFF% by more than 30% in the two-year period. All subjects underwent an oral glucose tolerance test and abdominal MRI to measure intrahepatic fat content (HFF%), visceral, and subcutaneous fat as previously described [Baseline fasting plasma GDF15 concentrations were measured in all subjects and in a subgroup of subjects with  according to manufacturer’s instructions. The detection threshold for this kit is 4.39 pg/mL and the assay range is 23.4–1,500 pg/mL. Plasma glucose was measured at bedside during the OGTT using the YSI2700-STAT-Analyzer , plasma insulin was measured by antibody radioimmunoassay from Millipore Sigma , and lipid levels were measured using Auto-Analyzer .χ2 test. Continuous variables are presented as mean (standard deviation) or as median (interquartile range) for HFF%. Non-normally distributed variables were log-transformed to approximate normality, except for HFF% that was transformed into rank (PROC RANK). A Mann-Whitney test was used to compare the differences between subjects with and without NAFLD and to compare differences between the cross-sectional cohort and the subgroup for longitudinal follow-up. A general linear model (PROC GLM) was used to test differences between NAFLD groups, and age, sex, ethnicity, BMIz, and visceral adipose tissue (VAT) were used as covariates. A GLM was also used to assess the association of GDF15 with VAT, using age, sex, ethnicity, BMIz, and HFF% as covariates. A simple regression analysis was used to test the association between change in intrahepatic fat content and change in GDF15 concentration. Statistical significance was established at an alpha of 0.05. Analyses were performed using SAS 9.4  and GraphPad Prism software 9.0.0 .Categorical data are presented as counts (percent) and were analyzed using a P = 0.224) and sex (P = 0.174). BMI z-score, measures of glucose metabolism and insulin sensitivity, and ALT levels were higher in the group with NAFLD. Fasting plasma GDF15 levels were significantly different between the two groups, independent from age, sex, BMI z-score, ethnicity, and VAT (P = 0.002)   Fig. . No assoex P = 0.74. BMI zP = 0.008), but after adjustment for intrahepatic fat content, the association became marginally significant (P = 0.042).GDF15 was associated with VAT independent of age, sex, ethnicity, and BMIz (p = 0.006), which disappeared after adjusting for HFF% (P = 0.082).Similarly, there was also an association between WBISI and GDF-15 independent from age, sex, ethnicity, and BMIz (P = 0.971).We measured plasma GDF15 during a 3-h OGTT and observed a slight reduction in GDF15 concentrations from 0 to 60 min. The GDF15 concentrations tended to have a greater initial reduction (0–60 min) and later rebound (60–180 min) in the group without NAFLD compared to the group with NAFLD Fig. . The delP = 0.008; r2 = 0.288)   Fig. . Similar69) Fig. .Fig. 2ChIn the present study, we observed an association between plasma concentrations of GDF15 and NAFLD and an association between changes in HFF% and changes in plasma GDF15 concentrations. Specifically, in subjects who experienced a reduction of NAFLD severity, plasma concentrations of GDF15 decreased, while, in those in whom the disease progressed, plasma concentrations of GDF15 increased significantly. Consistent with what has been shown before in adults, GDF15 concentrations in plasma tend to decrease during an OGTT and return to the initial concentrations by the end of the 3-hour study . Other sIt is likely that the upregulation of GDF15 might be a protective mechanism against the inflammatory response induced by free fatty acids in the context of fatty liver disease and the inflammation sustained by macrophages that leads to NASH and fibrosis. Moreover, Luan et al.  also shoWe acknowledge that our study has some limitations. The main limitation of the study is that, although the association observed between GDF15 and hepatic fat is strong, this study does not include information about liver histology that would be needed to understand whether GDF15 is related to changes in liver histology such as inflammation and fibrosis. Moreover, we are not able to explain whether this association underlies a causal relationship, or that it is merely an epiphenomenon within the context of more complex metabolic changes. Therefore, larger longitudinal and interventional studies are warranted to shed light on the metabolic dynamics linking GDF15 and NAFLD in youth with overweight or obesity.Supplemental Figure 1


Gomphocarpus fruticosus extracts in experimentally induced nephrolithiatic rats.In Ethiopian folk medicine, there is a claim that medicinal plants can treat urolithiasis although there is insufficient scientific evidence. The objective of this study was to evaluate the curative efficacy of Urolithiasis was induced in male Wistar rats by feeding ethylene glycol in drinking water for 28 days. The curative effects were evaluated after oral administrations of 200 mg/kg of the extracts from 15 to 28 days. Urine samples were collected 1 day before sacrificing the rats. Blood, liver and kidney samples were gathered under anaesthetic condition at day 28. Crystals in the urine were also analyzed by light microscopy.G. fruticosus EtOAc extract reduced significantly the level of sodium (P < 0.001), whereas it was significantly elevated the levels of magnesium and citrate (P < 0.01) compared to lithiatic control. G. fruticosus BuOH extract lowered the levels of potassium (P < 0.01), calcium and phosphate in urolithiatic rats. It was also observed that G. fruticosus EtOAc extract decreased the level of oxalate in the urine (P < 0.001), whereas it was increased the levels of magnesium (P < 0.05) and citrate (P < 0.01) in serum analysis after exposure to BuOH extract. In the kidneys, CaOx crystal deposits were reduced significantly by G. fruticosus EtOAc extract (P < 0.01).G. fruticosus EtOAc extract was potent in treating urolithiasis. However, further study is required to assess the efficacy of the active compounds against urolithiasis.It has been noted that  Urolithiasis (stones in the urinary tract) affects about 12% of the world population at some stage in their lifetime . UrolithRenal stone formation requires persistent retention of crystals in the kidneys after the completion of the crystallization process , 12. TheAn imbalance between urinary stone inhibitors and promoters has been suggested to be the cause of stone formation . PromoteDespite considerable improvements in medical therapy such as the utility of extracorporeal shock wave lithotripsy (ESWL), there is no satisfactory drug to treat renal calculi . ESWL trG. fruticosus (L.) Ait.f were selected to examine their effects on experimentally induced urolithiasis. A thorough literature survey was carried out to ascertain that none of the selected medicinal plant parts have been studied so far on anti-urolithiatic activities. To date, there is also insufficiency of scientific evidence reported on the antiurolithiatic activity of this medicinal plant. Therefore, the objective of the study was to evaluate the curative efficacy of medicinal plants in experimentally induced nephrolithiatic male rats.In Ethiopia, most patients rely on traditional medicinal plants as an alternative therapy for various diseases including urolithiasis. Medicinal plants are affordable, accessible, effective, with less side effects and being better compatible with the human body , compareGomphocarpus fruticosus leaves were collected during its flowering or fruiting times. The G. fruticosus (L.) Ait.f, known as Tifriena , was collected at Bole Bulbula around “93 Mazoria” Addis Ababa during October 2018. G. fruticosus L.Ait.f (Asclepiadaceae) (reported by key informant) was claimed for urolithiasis treatment. The plant specimens were submitted to the National Herbarium, Department of Plant Biology and Biodiversity Management, Addis Ababa University (AAU) for taxonomic authentication (TA238) and the corresponding collection number was given. The specimens were deposited in the National Herbarium of AAU for future references.The plant materials were cleaned thoroughly with tap water to remove contaminants, and dried in shed at room temperature from 2 to 3 weeks in the Biomedical Sciences laboratory, AAU. The dried plant parts were finely powdered using a kitchen grinder . The powders were put through a sieve of 3 mesh sizes so as to filter a gross solid matter.The extracts were prepared using a procedure similar to that often used by traditional healer’s or patients, with some minor modifications. The plant powders were soaked in distilled water, which was placed on a shaker for 72 h. The mixture was filtered through cotton/gauze, then through Whatman filter paper number 1 (pore size: 11 μm) to remove fine solid plant particles or insoluble constituents. The entire extracts were concentrated to dryness using lyophilizer machine by removing of distilled water under reduced pressure. Then, the semi-solid concentrates poured into a glass petri-plates and allowed to completely dry in water bath adjusted to 45 °C. The final dried extracts were collected and stored in labeled sterile bottles covered with tightly stopper and kept in freezer on -20 °C until used in the experiments.Gomphocarpus fruticosus mother extract (aqueous) was partitioned with solvents of increasing polarity from non-polar to polar solvents ensuring that a wider polarity range of compounds could be extracted. Successive extraction was sequentially started with petroleum ether, followed by chloroform, ethyl acetate, butanol and water, in which these solvents were removed and concentrated under rotary vacuum evaporator. The plant materials macerated in distilled water were lyophilized under the freeze dryer to obtain the powdered residues. The dried extracts were capped tightly with stoppered and stored in glass bottles, and kept in a refrigerator at -20 °C until used for the experiment.4H11NO3), and sodium chloride (NaCl) were purchased from the Micron International Trading House PLC (Addis Ababa Ethiopia). Similarly, isoflurane, and formaldehyde were purchased from Neway Chemicals PLC . Ethylene glycol (EG) and ammonium chloride (NH4Cl) were purchased from Pharma PLC . EDTA tubes, serum separator tubes, capillary tubes and surgical blades were purchased from Micro Pharma PLC . Furthermore, kits for liver and kidney function tests were purchased from a Roshi PLC . The oxalate  colorimetric assay kits and citrate colorimetric/fluorometric assay kits were purchased from BioVision Incorporated . Potassium citrate powder was obtained from the Black Lion Referral Hospital . Cystone  was purchased from Mumbi, India. Ascorbic acid, which was purchased from Micron International Trading House PLC .The chemicals and reagents or kits used were analytical grade purchased from various sources. The procurement of petroleum ether, chloroform, ethyl acetate, butanol and ethanol were from Wisteam PLC , potassium dihydrogen phosphate (anhydrous), sodium phosphate (dibasic anhydrous extra pure), Tris-HCl buffer  were used, purchased from the Ethiopian Public Health Institute (EPHI), and breed at the Biomedical Sciences animal laboratory, AAU. The experiment was conducted at the Biomedical Sciences Laboratory of AAU and EPHI, in accordance with internationally accepted standard guidelines for the use of animals in scientific research. Prior to starting the experiment, rats were acclimatized to standard laboratory conditions (6 rats per polypropylene cages) for 7 days. They were kept under a controlled environment of temperature (27 ± 2 °C), relative humidity (55 ± 5%), and light (12 h light/dark cycles). These rats were fed with regular pellets (standard diet) and allowed for free drinking water (ad libitum) for 28 days.4Cl) administered in the rats’ drinking water. In this hyperoxaluria model, 1% (w/v) NH4Cl was given with 0.75% (v/v) EG for the first 5 days to accelerate lithiasis, following this the water supply was switched to 0.75% EG alone for the next 25 days [Kidney stones were induced using ethylene glycol (EG) along with ammonium chloride (NH 25 days –47. Expo 25 days . EG admi 25 days . The expG. fruticosus PET extract), Group VI (G. fruticosus Chl. extract), Group VII (G. fruticosus EtOAc extract), Group VIII (G. fruticosus BuOH extract), and Group IX (G. fruticosus aq. fraction). Other investigators were also used potassium citrate (2.5 g/kg) as a positive control [In the curative treatment (dissolution), a total of 54 albino Wistar male rats were divided randomly into 9 groups comprising 6 individuals per group with matching body weights. In curative treatment, the disease was induced priory from day 1 to 14. Then, each of the extracts/drugs was administered orally from day 15 to 28 concurrently with disease induction (EG) protocols to determine curative effects , 51. At  control , 53. TheUrine, serum and histological profiles are indicators of stone formation as well as recurrences. The abundance and morphology of calcium oxalate crystals formed under in vivo stone inductions were examined using a light microscope .Rats were placed in separate metabolic cages and subjected to 24 h urine collection at day 28th for curative test . In crysAt the end of the respective treatment periods, the animals were individually housed in metabolic cages, and 24 h urine (acidified and non-acidified) samples were collected. Urine was acidified with 1 ml of 6 N HCL (hydrochloric acid) and stored at 4 °C for 5 days. Then, these were centrifuged at 3000 rpm for 10 min , and the supernatant of acidified urine were used to estimate excretions such as oxalate, calcium, magnesium and phosphate contents. In non-acidified urine sample contents such as citrate, creatinine and uric acid, and total protein concentrations were analyzed using commercially available diagnostic kits by the Automated clinical chemistry analyzer. Both oxalate and citrate concentrations were analyzed using multi-well spectrophotometer (ELISA reader) as per the manual provided with kits. For the purpose of quantifying, a calibration curve was prepared using Oxalate and Citrate kits as standard.After the end of the experimental period (day 28th), rats were anesthetized using Isoflurane and 3 ml blood samples were collected from the retro-orbital vein by capillary puncturing. Serum was separated after centrifuged at 3000 rpm, 20 °C for 15 min. The collected serum was investigated for biochemical parameters like creatinine, blood urea nitrogen, uric acid, urea, calcium, magnesium, and phosphate by Clinical Chemistry Auto-analyzer  with the respective diagnostic kits. The oxalate and citrate concentrations were determined using multi-well spectrophotometer (ELISA reader) as per the manual provided with kits. For the purpose of quantifying, a calibration curve was prepared using Oxalate and Citrate kits as standard.At the end of the experimental period (day 28th), rats were sacrificed under Isoflurane anesthesia, followed by cervical dislocation. Then, the abdomen was opened and both kidneys collected from each rat. The isolated kidneys were carefully removed, and cleaned (washed/rinsed) from extraneous tissues with an ice cold physiological saline solution (0.15 M NaCl). The left kidneys from each animal was preserved in 10% buffered neutral formalin and used for histological studies. The right kidneys were sliced into two equal halves using a blade, and one-half dried at 80 °C in a hot air oven. The method used was similar to Ashok et al.  with som-W180 10.1MP with 3x optical Zoom, New Jersey, USA) for further references.Histopathological examinations were done for kidney tissues of the experimental rats. All rats were sacrificed in a humane manner using Isoflurane anesthesia at the end of the 28th day (urolithiasis curative studies). The tissue pieces were taken from kidneys and analyzed for urolithiatic curative potentials of plant extracts. The tissues were fixed by 10% buffered neutral formalin solution, and subsequently embedded in paraffin wax. The sections (5 μm thick) were cut using Rotary Microtome 4060E (Germany), and mounted on glass slides and stained with Hematoxylin and Eosin to study the histopathological changes . All fieIn the kidneys, crystal depositions were determined using semi-quantitative assays, which is a microscopic scoring method , 56. TheIn counting CaOx crystal deposits, a sagittal section of each renal specimen was divided into four equal sized regions by two virtual lines, and readings of an average of 4 microscopic fields were reported , 58. A fP < 0.05 was considered statistically significant.The data were analyzed using Graph Pad Prism version 6 Software . One-way analysis of variance (ANOVA) followed by post-hoc Dunnett’s test comparisons were performed when necessary to compare between treated and untreated groups. The data values were expressed as mean ± standard deviation (SD). Values of In the curative (therapeutic) studies, the selected extracts dose was 200 mg/kg body weight of rats, which was one-tenth of the maximum tolerated dose 2000 mg/kg b.w . This waG. fruticosus successive solvent extracts, G. fruticosus EtOAc extracts . The level of potassium was reduced by G. fruticosus BuOH extract (P < 0.010). Treatment with the aqueous extract of EtOAc and BuOH extracts of G. fruticosus showed significant reductions in urinary protein excretions (P < 0.011) in relation to lithiatic control. Sodium concentration was also reduced very significantly with the treatment of EtOAc extract of G. fruticosus (P < 0.001). Treatment by the extract of G. fruticosus BuOH lowered the level of uric acid compared to lithiatic group (P < 0.050). However, lithogenic induction may cause impairments of renal function as evidenced by raising the levels of creatinine, proteins and uric acids. In the serum analysis, the level of potassium reduced by G. fruticosus EtOAc extract in comparison to lithiatic control (P < 0.010). It was also found that the effects of G. fruticosus BuOH extract on the level of chloride was close to the normal control. The level of creatinine raised by G. fruticosus crude extract and PET extract (P < 0.01) compared to the normal control . Similar to K-Cit, the effects of G. fruticosus EtOAc extract increased the level of citrate significantly (P < 0.010). G. fruticosus BuOH extract lowered the elevated levels of calcium and phosphate significantly in the urine compared to the diseased (lithiatic) control (Group II rats) (P < 0.010). It was also observed that G. fruticosus EtOAc extract decreased the level of oxalate in the urine compared to lithiatic control (P < 0.001). In the serum analysis, the excretions of calcium, oxalate and phosphate were grossly increased in lithiatic induced male Wistar rats. The EtOAc extract of G. fruticosus increased the concentration of magnesium (P < 0.052). The serum level of citrate was increased after exposure to BuOH extract of G. fruticosus (P < 0.011), suggesting similar effects to potassium citrate (P < 0.001). G. fruticosus Chl. extract significantly reduced the citrate level compared to the normal control (P < 0.001) (Table As indicated in Table 1) Table .Table 3P < 0.050). G. fruticosus PET extract increased serum AST level significantly (P < 0.001) compared to the healthy control. BuOH extract of G. fruticosus lowered significantly the level of AST similar to K-Cit (P < 0.001) compared to lithiatic control , and aspartate aminotransferase (AST) increased in the lithiatic control compared to the normal  control (ol Table .Table 4P < 0.050), except treatment with BuOH and Chl. extracts of G. fruticosus. The administration of the aqueous mixed extract and G. fruticosus EtOAc extract reduced the level of oxalate significantly compared to the untreated lithiatic groups (P < 0.011)  compared to the normal control (Group I). All treatments showed significant reductions in the level of phosphate P < 0.00, exceptG. fruticosus EtOAc extract (G), and its BuOH fraction (H) reduced the number of crystals compared to lithiatic control (B), but do not reduced for aqueous extracts of G. fruticosus (I)  Fig. .Fig. 2RG. fruticosus EtOAc extract (p < 0.010). The number of crystal deposits was counted via a sagittal section or longitudinal plane of each renal tissue specimen divided into 4 equal sized regions  and taking the mean of all fields . In contrast, previous studies reported that excessive excretions of sodium and chloride in the urine indicate the diuretic activity of the test extract [Urinary excretions of oxalate, phosphate, and calcium were increased, while the levels of magnesium and citrate levels reduced in the urine compared to the treated group at dose 200 mg/kg b.w, which were similar to the studies reported by Lemann et al.  and Sath extract , 66, wit extract .G. fruticosus EtOAc extract elevated the levels of magnesium and citrate (P < 0.010) similar to K-Cit in the curative test. The EtOAc extract of G. fruticosus decreased the levels of oxalate in the urine (P < 0.001) compared to lithiatic control in the curative study. In the curative test, significant (P < 0.011) reductions in urinary levels of calcium and phosphate following G. fruticosus BuOH extract administration indicates the presence of CaOx inhibitory constituents, which interfere with crystal nucleation and aggregations. Moreover, an increase in calcium concentrations is a favoring environment for nucleation, and precipitation of calcium oxalate or calcium phosphate with subsequent crystal growth [Magnesium binds with oxalate ions to form soluble oxalate complexes in the urine, and decreases the oxalate availability, which binds with calcium leading to CaOx formation , 69. Henl growth , 39. Siml growth , 70.G. fruticosus EtOAc extract (P < 0.010) reduced serum levels of potassium. In addition, the effects of G. fruticosus BuOH extract on the level of chloride was found to be close to the normal control during the curative study. The serum level of sodium was significantly reduced by G. fruticosus BuOH extracts (P < 0.050) compared to lithiatic control in the curative study. There was a decrease in serum oxalate excretions by EtOAc and BuOH extracts of G. fruticosus compared to lithiatic control. This might be either due to the inhibitions of oxalate formation, or interference with oxalate metabolism. Similarly, G. fruticosus EtOAc and BuOH extracts (p < 0.050) decreased calcium levels compared to lithiatic control in curative test. The ability of these extracts to alter calcium and oxalate excretions may be due to the disintegration of mucoproteins, which are promoters of crystallization as reported by Doddola et al. [G. fruticosus BuOH extracts (P < 0.011), suggesting a possible curative effect close to K-Cit treated group (P < 0.001). The presence of tannins and flavonoids can lead to the relaxation of smooth muscles of the urinary tract, which could facilitate the expulsion of kidney stones as studied in rats [G. fruticosus extracts in the present study.In curative study, a et al. . In the  in rats , which wG. fruticosus BuOH extracts lowered AST levels very significantly (P < 0.001), which was close to K-Cit compared to the lithiatic control in the curative studies. This can be attributed to the fact that abnormal levels of liver enzymes, particularly aminotransferases including alanine aminotransferase (ALT), and aspartate aminotransferase (AST) are prognostic features (indicators) of the damages of liver cells, and the cellular integrity of the kidneys [In the present therapeutic study, it was also evidently proved that the  kidneys , 72.G. fruticosus EtOAc extract (P < 0.051) and its BuOH extract (P < 0.051) treated groups, the severity of CaOx crystal depositions reduced the kidneys compared to lithiatic control, which was similar to a study on the other herbal extract [G. fruticosus reduced the level of phosphate significantly (P < 0.051) compared to the lithiatic control. Tissue injury could be caused by exposures to phosphate and calcium phosphate crystals, leading to the generation of oxidative stress, lipid peroxidation and depletion of antioxidant enzymes [Experimentally induced CaOx crystal deposition in the kidneys is also associated with localized inflammation as evidenced by infltration of monocyte and macrophages to the site . G. frut extract . In cura enzymes . Consequ enzymes , 76. COM enzymes . The pre enzymes .A major limitation of the study is the inability to use a polarized light microscope that magnifies and better resolve in vivo crystallizations to differentiate between COM and COD crystals which would have improved data quality. The effects of test extracts on urinary diuretic activities were not assessed. Despite these limitations, the strength of the present study was the use of standard experimental designs and procedures to test the potential efficacy of the traditional medicinal plants used in the study.G. fruticosus extracts in urolithiasis management. In vivo/animal tests confirmed the curative efficacies of G. fruticosus extracts, which were evidenced from biochemical and histopathological findings. The present findings demonstrated the CaOx curative efficacy of G. fruticosus EtOAc extract. Further research is necessary to identify the active compounds responsible for the treatment of urolithiasis.The present study has provided some evidences for the traditional use of


Peritoneal dialysis (PD) patients experience accelerated arterial aging, which is characterized by elastin degradation. Elastin-derived peptides (EDPs) are direct products of elastin fragmentation. This study tried to explore the association between serum EDPs and abdominal aortic calcification (AAC) in PD patients.Serum levels of EDPs were analyzed in 126 eligible PD patients and 30 controls. PD patients were grouped according to the annularity of AAC evaluated by an abdominal computed tomography (CT) scan. Serum EDPs were analyzed in relation to the presence of AAC or severe AAC in PD patients by logistic regression analysis.Serum EDPs in PD patients were significantly higher than age-matched controls. In 126 PD patients, higher EDPs was associated with greater risk of present AAC  and severe AAC . A combination of EDPs substantially improved the accuracy of diagnostic performance for AAC and severe AAC.EDPs can predict the presence and extent of AAC in PD patients, indicating its possible role to recognize PD patients at risk for AAC and severe AAC. Cardiovascular disease (CVD) remains one of the major causes of death in patients starting peritoneal dialysis (PD) , and thoVascular calcification is thought to be a process of ageing accompanied by obvious extra-cellular matrix remodeling featured by elastin degradation . ConcurrIn uremic patients with accelerated arterial aging, elastodystrophy of aorta occurred early and constructed a context of altered mineral metabolism for the pathogenesis of arterial medial calcification . In dialThe study included adult PD patients and healthy volunteers from August 2018 to January 2020 in the First Affiliated Hospital of Soochow University. All participants signed an informed consent, and the study was approved by the ethics committee in accordance with the Declaration of Helsinki.PD patients receiving continuous ambulatory peritoneal dialysis with low calcium (1.25 mmol/L) peritoneal dialysis solution  for at least three months were included. Exclusion criteria were (1) history of parathyroidectomy, (2) chronic obstructive pulmonary disease (COPD), (3) liver cirrhosis, (4) cancer, (5) peritonitis or infection within 1 month, (6) acute myocardial infarction or aortic dissection within 1 month, and (7) incomplete information on AAC evaluation. The flowchart was displayed in At the same time, 30 healthy subjects were randomly selected among age- and sex-matched people undergoing medical examination in the Medical Examination Center. Every of them had a normal glomerular filtration rate and underwent a CT scan to rule out the presence of AAC. They also had no history of proteinuria, hematuria, hypertension, or diabetes.3 and density ≥130 Hounsfield unit [All patients received noncontrast abdominal MSCT during hospitalization, and the detailed CT protocol was presented in Online Resource 1. AAC was defined as calcified plaques with area ≥1mmeld unit  along theld unit . Exampleg for 15 min at 4 °C. The upper serum was separated and stored at −80 °C. Until the investigation, serum samples were thawed at room temperature. Specific competitive enzyme-linked immunosorbent assay kits  were used to measure serum EDPs. The principle and assay procedures were summarized in Online Resource 1.Venous samples were collected for at least 10-h fast. After clotting for 30 min at room temperature, the samples were centrifuged at 4000 Age, gender, body mass index (BMI), dialysis vintage (PD vintage), weekly urea clearance , and blood pressure were recorded on purpose. At the same time, baseline comorbid conditions of hypertension, type-2 diabetes mellitus (T2DM), and CVD consisting of coronary artery disease, cerebral artery disease, and aortic aneurysm were abstracted from previous medical records, as well as the related medications.Hemoglobin (HB), neutrophil cells (NE), platelets (PLT), lipid profile, creatinine (Cr), β2-microglobulin (β2-MG), uric acid (UA), albumin (Alb), and high sensitivity-C reactive protein (hs-CRP), mineral metabolism parameters including calcium (Ca), phosphate, intact parathyroid hormone (iPTH) and alkaline phosphatase (ALP)  were obtThe continuous variables were expressed as mean ± standard deviation (SD) or median (Q1 – Q3). One-way analysis of variance (ANOVA) or Kruskal–Wallis H test, and independent sample T-test or Mann–Whitney test were used appropriately. Categorical variables were presented as frequencies (percentages) and compared by the Chi-square test or Fisher’s exact test when necessary. Spearman correlation analysis was used to find association of EDPs with other parameters.PD patients were firstly assigned into AAC versus non-AAC group, and a logistic regression analysis was conducted among PD patients to examine the relationship between EDPs with AAC. We also conducted a subgroup analysis in patients with AAC. Considering the imbalanced sample size, patients were further grouped into severe versus non-severe group with the cutoff annular of 90 degree, as shown in Finally, our study enrolled 126 patients on PD and 30 healthy controls. Patients and controls were comparable in age and sex ratio; however, serum EDPs in PD patients were significantly higher than that in controls. Hyperlipidemia was absent in PD patients, while disorders of mineral parameters were more prevalent (higher serum P and ALP). Detailed demographic and clinical characteristics of all PD patients are summarized in r = 0.511), annularity of AAC (r = 0.488), ALP (r = 0.382) and TC (r = 0.236) as shown in The mean age of all PD patients was 50 ± 13 years and the median dialysis vintage was 22.5 (7.0 − 51.0) months. Correlation analysis showed positive association of EDPs with age  patients were in the AAC group, and 41 (32.5%) were within the non-AAC group. As shown in Figure S2).ROC curves of EDPs and mineral bone disorder parameters showed that EDPs had the most excellent power to discriminate between PD patients with or without AAC. The area under the curve (AUC) of EDPs was 0.767, which was the largest , total triglyceride (TG), and history of T2DM were incorporated into the multivariate regression model. After fully adjusted, higher EDPs (OR = 1.056), along with higher age (OR = 1.097), and longer PD vintage (OR = 1.042) were associated with greater odds of AAC in PD patients. However, UA was also a significant determinant (OR = 0.987).To examine the association between serum EDPs and AAC in PD patients, eligible covariates with ROC curve analysis showed that the cutoff point of EDPs to predict AAC was 43.22 ng/mL, and the sensitivity and specificity values were 74.1% and 70.7%. When EDPs were transferred to a categorical variate according to the cutoff point, OR of EDPs increased to 2.810 . A combiTable S2).Of all 85 PD patients with AAC, 36 (42.35%) patients were categorized into the severe group with AAC annular > 90 degree, and 49  (57.65%) were in the nonsevere one. When compared with each other, patients with severe AAC were older with longer PD vintage and higher EDPs. Noticeably, classical risk factors of vascular calcification such as phosphate, iPTH and hs-CRP were significantly higher in severe AAC group. They also tended to have higher uremic toxicity biomarkers of β2-MG and Cr . At the As shown in The evaluation of cardiovascular calcification has been the focus in dialysis patients as it predicts a higher risk of CVD . PD patiElastin degrades irreparably with aging, and circulating concentration of EDPs increased steadily throughout the human life span . PatientElastin degradation participates in the pathogenesis of atherosclerosis and medial calcification ,26, bothThe strongest correlation of age with increased EDPs in PD patients of our study also suggested that elastin degradation might also be age-related, but further accelerated in those patients . Activatet al also found higher SUA was associated with lower cardiovascular and all-cause mortality in 492 PD patients [Vascular calcification is such a complex process that other factors participating it should also be taken when evaluating the risk of AAC. We found that longer dialysis vintage played an important role in both AAC and severe AAC of PD patients. This was identical to several larger studies, in which duration of dialysis was related to premature AAC evaluated by lateral lumber radiography or conduit arteries medial calcification detected by ultrasound . Besidespatients . While apatients . The rolin vitro and in vivo studies are needed to elucidate its relationship with AAC in PD patients.At the same time, there exist several limitations in our study. First, this is a cross-sectional study with a small sample size. The association of EDPs and vascular calcification needs further validation in future multicenter researches. Second, we cannot figure out the causal link between EDPs and AAC progression because of a lack of following up. Finally, the model was based on limited clinical information. EDPs are biologically active and can interact with various cells and participate in physiological and pathological processes , thus inIn conclusion, EDPs improved the risk stratification of AAC and severe AAC among PD patients and provided novel insights into EDPs on AAC as a potential indicator.Click here for additional data file.


He had a twin brother who had normal auxological/pubertal development . Anorexia nervosa was diagnosed, and he was enrolled in a personalized treatment and surveillance program. “Nonthyroid illness” resembling secondary hypothyroidism was noted, as was low bone mineral density. Clinical and biochemical follow-up showed significant improvements in BMI , completion of puberty (testicular volumes: 25 mL), and reversion of main neuroendocrine abnormalities. Herein, we present an adolescent male with arrested puberty in the context of anorexia nervosa. The recognition of this rare condition in males allows a personalized approach to disordered puberty, with resumption of normal function of the hypothalamic-pituitary-gonadal axis and achievement of pubertal milestones.The normal development of puberty depends on the specific pulsatility of gonadorelin, which is finely regulated by genetic and environmental factors. In the published literature, eating disorders figure as a cause of pubertal delay/arrest in females but are rarely considered in males with disordered puberty. A 16.7-year-old male was referred to the Department of Pediatrics with arrested puberty due to severe malnutrition in the context of food restriction. Past medical history was relevant for asthma. Generalized cachexia, facial lanugo hair, cutaneous xerosis, and Russell's sign were noted; he had a height of 155.5 cm  and a BMI of 12.4 kg/m Puberty is the process of maturation of the hypothalamic-pituitary-gonadal (HPG) axis that culminates in the full achievement of final height, secondary sexual features, and fertility capacity . This no2 (−6.8 SD); his height chart showed a progressive decrease in his linear growth, markedly (height velocity: 2.7 cm/year) after an age of 13 years  and resumed puberty (testicular volumes: 12 mL). General biochemistry showed correction of initially abnormal parameters , without imaging evidence of vertebral fractures. Bone age was estimated to be delayed by 8 months relative to chronological age. Currently, at 17.6 years old, he maintains his follow-up with a multidisciplinary team and his personalized nutritional plan, including daily supplementation with vitamin D (800 UI/day) and calcium (1200 mg/day). His height had a slight increase to 157.4 cm (−2.2 SD), as was his BMI to 16.2 kg/m2 (−2.55 SD). Testicular volumes increased to adult size (25 mL). Basal endocrine reevaluation showed increased total testosterone levels and correction of “nonthyroid illness”; high late-night salivary cortisol was noted , weight was 30.2 kg, and BMI was 12.5 kg/mears see ; his BMIears see ; the leframeters ; basal erameters . Dual-enas noted .Herein, we present a rare case of an adolescent male with arrested puberty due to severe undernutrition in the context of AN, which, with personalized treatment, could successfully resume his puberty. Data are lacking regarding the pubertal status (Tanner stage) of patients with AN, both at diagnosis and following treatment institution . AvailabMalnutrition-induced hepatitis is common among individuals with AN , 28 as aZ-scores <−2 SD and bone fractures) [Our patient had low BMD. Adolescence is a period of increased bone accrual towards the attainment of peak bone mass, which is markedly impaired in adolescents with AN , 31, andactures) .In conclusion, the presented case illustrates the importance of identifying the underlying aetiology of arrested puberty and a focused multidisciplinary approach to revert functional abnormalities of the HPG axis that allow pubertal resumption and completion. Early diagnosis and treatment are crucial to blunt the drivers of neuroendocrine abnormalities characteristics of AN, which may lead to suboptimal height gain, unachieved pubertal milestones, and a BMD prone to fractures.


To evaluate the clinical management of choanal atresia (CA) in tertiary centers across Canada.Multi-centre case series involving six tertiary care pediatric hospitals across Canada. Retrospective chart review of patients born between 1980 and 2010 diagnosed with choanal atresia to a participating center.2 (2) = 1.6, p = .46].The health charts of 215 patients  with choanal atresia (CA) were reviewed. Mean age of initial surgical repair was 0.8 months for bilateral CA, and 48.6 months for unilateral CA. Approaches of surgical repair consisted of endoscopic transnasal (31.7%), non-endoscopic transnasal (42.6%), and transpalatal (25.2%). Stents were used on 70.7% of patients. Forty-nine percent of patients were brought back to the OR for a planned second look; stent removal being the most common reason (86.4%). Surgical success rate of initial surgeries was 54.1%. Surgical technique was not associated with rate of restenosis [χThe present study is the first national multi-institutional study exploring the surgical outcomes of CA over a 30-year period. The surgical repair of CA presents a challenge to otolaryngologists, as the rate of surgical failure is high. The optimal surgical approach, age at surgical repair, use of stents, surgical adjuncts, and need for planned second look warrant further investigation. Choanal atresia (CA) is a congenital condition resulting in obstruction of the posterior nasal passage(s), known as the choana, with an incidence of approximately one in 5000 to 8000 live births , 2. ManaTo date, best practices for management of CA remain to be determined. Method of surgical correction of CA is usually dictated by surgeon or institutional preference . PostopeThe reasons for these unresolved controversies regarding best practices for clinical management of CA is partly due to the rarity of the disorder and consequently the limited level of evidence available in the literature. Published studies often report case series of single surgeon or single institution experiences, involving small sample sizes –11, withThe aim of the current study is to elucidate controversies surrounding the management of CA using a multi-institutional approach. The Canadian landscape lends a unique setting to evaluate the management of CA, as all CA repairs are performed in academic settings. In a two-part study, this national investigation provides a comprehensive review of the clinical presentation (Part I) and management (Part II) of a large sample of patients with CA treated at tertiary care centres across Canada. The present paper reports on the clinical management of CA. Specifically, the following outcomes were explored: surgical management and use of adjunct procedures, utilization of a post-operative planned second look, and success rate of surgical repair for CA.Pediatric otolaryngologists practicing in tertiary care centers across Canada were invited to take part in the study via phone or email. Of the nine healthcare centers who were contacted, six centers  agreed to participate in the chart review. Health records of patients born between 1980 and 2010 diagnosed with CA who underwent treatment at participating centers were included in this study. Patients born before 1980 or after 2010 or patients who did not receive a definitive diagnosis of CA were excluded from the study. A standardized checklist was completed for each patient presenting with CA who met study inclusion/exclusion criteria. The following variables were collected from patient chart review: surgical approach, surveillance, surgical instruments, adjuncts to surgery, findings of planned second look, revision surgeries, reasons for revision surgeries, and, a comparison of surgical revision rates between surgical approaches, and for bilateral versus unilateral cases of CA. In the current sample, surgical success was defined as presence of choanal patency, with no need for surgery for restenosis or debridement of granulation tissue after initial repair. Each participating center obtained ethical approval for this study from their institution’s Health Research Ethics Board.A descriptive analysis of study outcomes is presented. In addition to descriptive statistics, results of chi-square tests/Fisher’s exact tests, independent samples t-tests, and Spearman rho correlation coefficients were reported where appropriate. Statistical tests were conducted with SPSS . Alpha was set a priori at .05 to determine statistical significance.n = 26), Toronto (n = 83), Edmonton (n = 17), Vancouver (n = 60), Montreal (n = 11), Halifax (n = 18)] met the study inclusion/exclusion criteria and were included in this national study. Health records of 215 patients were reviewed. One-hundred and twenty-seven patients (59.6%) were female and 88 (40.9%) were male. The mean age of patients at time of CA presentation was 0.4 months (range 0.1 to 7.2 months) for bilateral CA and 37.8 months (range 0.1 to 164.1 months) for unilateral cases.Two-hundred and fifteen patients across the six participating centers .Stents were used in 70.7% of patients (2(1) = 17.7, p < .001]. Patients with bilateral CA also had their stents inserted for an average of 1 month longer than patients with unilateral CA .Patients with bilateral CA were significantly more likely to have stents placed compared to patients with unilateral CA .The median number of revision surgeries was 1 with a range of 0 to 8 revision surgeries for the study cohort. Table 2(1) = 17.7, p < .001]. Patients with bilateral CA also had their stents inserted for an average of 1 month longer than patients with unilateral CA .Table 2(1) = 2.8, p = .10]. However, patients with bilateral CA were significantly more likely to require a 2nd and 3rd revision surgery  than patients with unilateral CA. Individuals with unilateral CA had a median of 1 revision surgery (range 0 to 3 revision surgeries), while individuals with bilateral CA had a median of 2 revision surgeries (range 0 to 8 revision surgeries).Patients with bilateral CA were not significantly more likely to require a 1st revision surgery than unilateral CA  and use of adjunct procedures. As this was a retrospective study, these patient and treatment factors could not be accounted for. To date, only two small randomized controlled trials have been conducted on CA; one evaluating Mitomycin-C use versus no-use in 20 children who were treated with the ETN approach  and one Furthermore, three tertiary care centres in Canada did not participate in the present investigation, limiting the generalizability of study findings. Finally, given the multi-institutional nature of this study, results of the study may be confounded by institutional factors including variability in surgeon expertise, institutional preferences for use of different surgical techniques, time of repair and use of adjunct procedures. This variability across sites may limit the interpretability of study findings.This retrospective multi-center investigation reported on over 30-years experience of patients with CA across tertiary care centers in Canada. Surgical repair type, whether it be ETN, NTN, or TP was not statistically significantly associated with rate of restenosis, while a weak negative association between age and number of revision surgeries was found. Stenting was used often in this patient cohort and stent removal, repositioning, and replacement were the primary reasons for a planned second look in the OR following initial surgical repair. Laser use facilitated NTN approaches most frequently, with limited use in ETN and no use in TP approaches. Mitomycin-C was used infrequently in the present patient cohort. Future investigations regarding hereditary linkages, the need for a planned second look, stenting, laser and Mitomycin-C use, and surgical repair approaches are warranted.


C-reactive protein (CRP) is one of the most common oxidative indexes affected by many diseases. In recent years, there have been many studies on CRP, but the relationship between CRP levels and the cardiovascular risk in the Chinese young female population is still unclear. The purpose of this work is to explore the predictive value of CRP for the cardiovascular risk in the Chinese young female population. n = 402); subgroup 2: 1.0 mg/L ≤ CRP ≤ 3.0 mg/L (n = 303); subgroup 3: CRP > 3.0 mg/L (n = 135). The levels of CRP were observed in the two groups and three subgroups.  The study is conducted by 1 : 1 case-control to retrospectively analyze 420 young women with acute coronary syndrome (ACS group) who underwent percutaneous coronary intervention (PCI) and 420 young women (control group) who underwent coronary angiography (CAG) to exclude coronary heart disease from January 2007 to December 2016. All patients are divided into three subgroups according to CRP values: subgroup 1: CRP < 1.0 mg/L (p < 0.001), and patients with higher CRP levels were associated with a significantly increased rate of major adverse cardiovascular events (MACE) . After adjustment for baseline covariates, CRP level was still an independent predictor for the incidence of MACE, either as a continuous variable or as a categorical variable. There was a significantly higher rate of all-cause mortality and myocardial infarction in patients with higher CRP values during follow-up.  A total of 840 patients were analyzed. The mean duration of follow-up was 66.37 ± 30.06 months. The results showed that the level of CRP in the ACS group was significantly higher than that in the control group (1.30 ± 1.70 vs. 3.33 ± 5.92, respectively,  The research results show that high CRP is associated with increased risk of ACS in the Chinese young female population. Risk stratification with CRP as an adjunct to predict clinical risk factors might be useful in the Chinese young female population. Coronary heart disease (CHD) has become one of the top leading causes of death among Chinese adults . The carn = 402); subgroup 2 (average risk): 1.0 mg/L ≤ CRP ≤ 3.0 mg/L (n = 303); and subgroup 3 (high risk): CRP > 3.0 mg/L (n = 135). The levels of serum CRP were observed in the two groups and three subgroups. Baseline characteristics of the enrolled patients were recorded in detail, including age, gender, body mass index (BMI), smoking history, hypertension, diabetes, the level of CRP, and other biomarkers. Exclusion criteria included the following: including congestive heart failure (CHF), myocardiopathy, congenital heart disease, history of aorta surgery, rheumatic heart disease (RHD), severe valve disease, pulmonary or connective tissue disease, active cancer, infective endocarditis, and chronic kidney disease. All the patients were recruited from the Chinese population, and all subjects signed the informed consent. All patients with ACS took prescribed dual antiplatelet therapy, including aspirin  and clopidogrel  for at least 12 months after PCI regardless of drug-eluting stent (DES) type. Individual medical management decisions during hospitalization were exclusively decided by their responsible interventional cardiologists and physicians. Clinical follow-up was performed by telephone contact or outpatient clinical visits. The date of follow-up was ended in December 2018. The primary endpoints for this study were MACE, which included composite occurrence of cardiac death, nonfatal myocardial infarction, and target vessel revascularization (TVR). The follow-up time range was from 11 months to 146 months. The mean duration of follow-up was 66.37 ± 30.06 months.The study was conducted by 1 : 1 case-control to retrospectively analyze 420 young women with ACS (ACS group) who underwent PCI and 420 young women (control group) who underwent coronary angiography (CAG) to exclude CHD by propensity score matching from January 2007 to December 2016. All patients were divided into three subgroups according to risk stratification from the American Heart Association (AHA): subgroup 1 (low risk): CRP < 1.0 mg/L (t-test. The counting data were expressed as a percentage (%), and the chi-square (X2) test was used for comparison between groups. The association of CRP and MACE was investigated by multivariable Cox proportional-hazard models adjusted for all potential confounding factors. The test level was set as a double-tail test a = 0.05, p < 0.05 was statistically significant, and p < 0.01 was statistically very significant.The sample size of this study was calculated by using Power Analysis and Sample Size software . The data statistical analyses were performed using the Statistical Package for Social Sciences software . All data of cases were inputted into the computer software database. The continuous variables with normal distributions were expressed as mean ± standard deviation. The comparisons between groups were performed using the independent Student's p > 0.05). Compared with the control group, the proportion of overweight, smoking, hypertension, diabetes, and hypercholesterolemia in the ACS groups is significantly higher (p < 0.05). The level of CRP in the ACS group is significantly higher than that in the control group .There is no significant difference in age between the two groups (p < 0.05).The result suggests that CRP was an independent predictor of ACS . Overweight, smoking, hypertension, diabetes, hypercholesterolemia, unstable angina, double coronary lesions, three coronary lesions, total stent length, SCr, Uric, LDL, HDL, TC, and TG in subgroup 3 were significantly higher than those in subgroup 1 and subgroup 2 (p < 0.05).There was no significant difference in age, HB, and Hcy in the three subgroups (Patients with higher CRP levels (CRP > 3 mg/L) were associated with a significantly increased rate of MACE. Cox regression model analysis illustrated that there were significant increases in MACE in patients with higher CRP during the follow-up see .p < 0.001). The result of bivariate logistic regression analysis of the relationship between CRP and ACS showed that CRP was an independent predictor suffering from ACS . Patients with higher CRP levels (CRP > 3 mg/dL) were associated with a significantly increased rate of MACE which included all-cause death, recurrence of nonfatal myocardial infarction, and target vessel revascularization (TVR) in terms of long-term clinical outcome. Cox regression model analysis illustrate that there was a significant increase rate of MACE in patients with rising CRP during the follow-up. Among the three CRP subgroups, the event-free survival curve was different, and the survival curve of subgroup 3 was significantly lower than that of the other two groups. We believe that CRP, as a useful and easily available biomarker, can be used to improve the risk assessment and secondary prevention of coronary heart disease. At the same time, it was found that there was a good correlation between the CRP level and the severity of coronary artery lesions. CRP level was an independent predictor for the incidence of MACE. Compared with the lower CRP, the patients with higher CRP values had a significantly higher rate of MACE during follow-up [CRP is an acute inflammatory response protein induced by cytokines and is secreted by the liver and activated macrophages in atherosclerotic plaques . CRP alsollow-up .Our research indicates that high CRP is associated with increased risk of ACS in the Chinese young female population. Risk stratification with CRP as an adjunct to predict clinical risk factors might be useful in the Chinese young female population.