Rotator cuff injury is the most common cause of shoulder dysfunction. Despite the continuous advancement of surgical techniques, the incidence of re-tearing after rotator cuff repair is still high. The main reason is that it is difficult to reconstruct the normal tendon bone interface and the process is slow, and the application of tissue engineering technology can promote tendon and bone healing. This study will evaluate the effect of the bionic double membrane stent on the rotator cuff healing after arthroscopic rotator cuff repair.This is a prospective randomized controlled trial to study the effect of biomimetic double-layer biofilm stent on rotator cuff healing. Approved by the clinical research ethics committee of our hospital. The patients were randomly divided into 1 of 2 treatment options: (A) a biomimetic double-layer biofilm stent group and (B) a non-bionic dual-layer biofilm stent group. Observation indicators include: visual analog scale score, University of California Los Angeles score, American Shoulder & Elbow Surgeons score and Constant-Murley score. Data were analyzed using the statistical software package SPSS version 16.0 .This study will evaluate and evaluate the effect of the bionic double-layer membrane stent on the rotator cuff healing after arthroscopic rotator cuff repair. The results of this experiment will provide new treatment ideas for promoting rotator cuff tendon bone healing.DOI 10.17605/OSF.IO/FWKD6 The treatment of rotator cuff injury is complicated, and 1-stage repair is feasible for patients who have failed non-surgical treatment. Arthroscopic rotator cuff repair has become the “gold standard” for rotator cuff injury repair, and has the advantages of less trauma and fewer complications.Rotator cuff tear refers to the continuous interruption of cuff like structure formed by the tendons of supraspinatus, infraspinatus, subscapularis and teres minor, due to trauma, degeneration and strain, impact, blood supply damage and other reasons, which causes shoulder pain and limited movement as the main clinical manifestations. It mostly occurs in middle-aged and elderly people, with an incidence rate of 6% in the younger than 60-year-old age group, and 30% in the older than 60-year-old age group, There are many reasons for the failure. The main reason is that it is difficult to reconstruct the normal tendon-bone interface and the process is slow. Compared with the normal fibrocartilage bone-tendon junction, the scar connection formed after traditional surgery has significantly lower strength, Although this kind of scar can reshape the fibrocartilage structure, the process often takes as long as 6 months to 1 year. In order to make postoperative tendon healing more firm and rapid, it is necessary to find and apply new methods to provide an ideal condition to improve the activity of tissue cells around the rotator cuff healing site and rebuild the biomechanical function of the tissue.With the deepening of the understanding of the disease and the development of surgical techniques, the rotator cuff repair has achieved satisfactory clinical results, but the postoperative imaging follow-up found that the success rate of biological repair of anatomical structures is low.–10 The biomimetic double-layer biofilm scaffold is a biomimetic periosteum that combines growth factors and bioactive stem cells. It controls the release of bone morphogenetic protein 2 (BMP-2) and vascular endothelial growth factor (VEGF) and platelet derived growth factor (PDGF) form specific expression, combined with the differentiation of bone marrow mesenchymal stem cells to help the regeneration of cartilage at the tendon-bone interface and promote tendon-bone healing.At present, biological methods and materials based on growth factors, repair scaffolds, and stem cells have become the research hotspots of strategies to promote rotator cuff tendon bone healing.In this study, a randomized controlled trial will be used to observe the effect of the biomimetic double-layer biofilm stent on the healing of the rotator cuff after arthroscopic rotator cuff repair, and to evaluate its long-term efficacy through follow-up and explore its effectiveness in promoting tendon bone healing after rotator cuff repair.22.1 The flow chart is shown in Figure This is a prospective single-center randomized controlled trial to study the effect of a biomimetic double-layer biofilm stent on the healing of the rotator cuff after arthroscopic rotator cuff repair. The research protocol will follow the Declaration of Helsinki and comprehensive trial reporting standards.2.2This research program is in accordance with the Helsinki Declaration and approved by the Clinical Research Ethics Committee of our hospital. This protocol has been registered in open Science Framework (OSF) (Registration number: DOI 10.17605/OSF.IO/FWKD6). All patients need to sign a written informed consent before they are randomly assigned to continue the trial.2.3The calculation of the sample size is based on the visual analog scale (VAS)score results 1 month after the operation. According to the pre-test results, the average score of the experimental group is estimated to be 3.4 points and the standard deviation is 1.8. The control group averaged 4.6 and the standard deviation is 1.9. This study is a non-inverse design, 1-sided test, α =0.025, power = 90%, through calculation, each group needs 50 patients in total, and the estimated withdrawal rate is 20%. Each group will include 63 patients.2.4Inclusion criteria: ①Age 45 to 70 years; ②Shoulder joint pain, active flexion and lifting weakness, affecting work and daily life, non-surgical treatment for more than 6 months, and non-surgical treatment for more than 6 months; MRI examination confirmed full-thickness rotator cuff tear ③Patients undergoing arthroscopic rotator cuff repair for the first time.Exclusion criteria: ①Partial tear; ②Huge or irreparable tear; ③With nerve injury; ④Biceps long head tendon injury; ⑤Shoulder joint osteoarthritis, rheumatoid arthritis, etc.; ⑦ Those who have a history of shoulder surgery and a history of mental illness.All patients participating in the study will be informed of the method, purpose and possible risks of the study. At the same time, they need to sign a written informed consent. They are free to choose whether to continue the trial at any time.2.5Eligible participants were randomly assigned to the treatment group or the control group at a ratio of 1:1 using a random tool based on the central network. Randomization was performed without any stratification generated by independent statisticians who are not involved in trial implementation or statistical analysis using SAS 9.3 software . The clinical research coordinator enters the participant information on the tablet and Was given a random number. The research assistant gets the participant's assignment from the computer. Throughout the research process, the research assistant is responsible for screening, recruiting participants, and assigning random numbers to the participants who have been included. The result assessor is responsible for the assessment of the scale. Except for the surgeon, all doctors, anesthesiologists, research assistants, participants, intervention supervisors, and statisticians who conduct statistical analysis, the division of personnel is unknown.2.6(1)Arthroscopic evaluation: The glenohumeral joint will be examined through the posterior approach, and the rotator cuff joint will be examined through the anterior approach. The posterior approach is used to enter the subacromial space and the lateral approach was established. The shape of rotator cuff tear and the degree of tendon retraction are observed from the posterior and lateral approach respectively, and the rotator cuff tear is comprehensively evaluated. All patients are identified as full-thickness tears under arthroscopy without large and irreparable tears.(2)Conventional arthroscopic repair: The operation will be completed by the same surgeon. Under general anesthesia, the patient takes the lateral decubitus position, disinfects, spreads the towel, and traction of affected limb. After exploring the joint cavity through the conventional arthroscopic approach, the patient enters into the subacromial region. The patients with subacromial impingement sign are given conventional subacromial decompression, and the rotator cuff break and rotator cuff insertion bone surface are freshened under the microscope, and the rotator cuff tissue is repaired with single or double row wire anchors.(3)Implantation of the biomimetic double-layer biofilm scaffold: the bone surface of the damaged rotator cuff is freshened, and the membrane containing BMP-2 microspheres is placed on the humerus side, and the membrane containing VEGF and PDGF poly lactic-co-glycolic acid (PLGA) microspheres is placed on the tendon side.The experimental group underwent conventional arthroscopic repair after implantation of the biomimetic double-layer biofilm stent, and the control group only underwent conventional arthroscopic repair. There were no special differences in other consumables during the operation.2.7(1)VAS is often used to measure pain intensity. The patient will be asked to mark a point between 0 and 100, where 100 means maximum pain (far right) and 0 means no pain (far left);VAS:(2) The American Shoulder & Elbow Surgeons score includes pain (50 points) and life function (50 points). The full score is 100 points. The higher the score, the better the shoulder joint function;American Shoulder & Elbow Surgeons score.(3) Including pain, function, muscle strength and range of motion of the shoulder joint. The total score is 35 points, the higher the score, the better the shoulder joint function;University of California Los Angeles score:(4) It is the shoulder joint scoring method uniformly adopted by the European Society of Shoulder and Elbow Surgery. It consists of 4 subscales: pain (15 points), muscle strength (25 points), functional activity (20 points) and shoulder joint mobility (40 points). The total score is 100 points, the higher the score, the better the shoulder joint function.Constant-Murley score:(5)X-ray evaluation of postoperative shoulder joint anatomy, MRI evaluation of tendon anchor loosening and displacement and rotator cuff healing.(6)Incision healing grade, incidence of adverse reactions .2.8All patients undergo rehabilitation training according to the standard plan. Postoperatively, the shoulder joint fixation device is used to fix the affected shoulder at 30° internal rotation and 20° abduction. From 0 to 6 weeks after surgery, mainly passive activities, the main actions: pendulum, forward bend, lift and external rotation, forward flexion and abduction should not exceed 90°; active exercise will be started after 6 weeks; physical labor is allowed after 6 months or sports. After 3, 6, and 12 months outpatient follow-up, X-ray and MRI examination of the shoulder joint are performed.2.8.1During each follow-up, 1 to 2 assistants will collect the follow-up data according to the evaluation criteria, and record them clearly in a pre-designed form combined with the basic information of patients. The information and data of all patients will be collected and stored in a fixed storage room to protect the confidentiality before, during and after the trial. Access to the database is limited to the researchers of this research group.2.8.2P value of < .05 was considered statistically significant.We will use SPSS 16.0 statistical software for statistical analysis. Continuous variables were described as the mean ± standard deviation, and differences between groups were analyzed using a series of 1-way analysis of variance (ANOVA) with Bonferroni post-hoc test, while differences between groups over time were analyzed using multi-way ANOVA with Bonferroni post-hoc test. Categorical variables were described as the number (%), and were analyzed by Fisher exact test. A 3 Most scholars believe that poor healing of the tendon at the stop of the rotator cuff is a key factor affecting the effect of surgery. In recent years, with the rapid development of tissue engineering, the repair of the tendon-bone interface of rotator cuff injuries has also made great progress. Different treatment methods have their own advantages, and the combined application of multiple strategies may be the most effective method.At present, the incidence of rotator cuff nonhealing and re-tearing after rotator cuff injury is still high.The biomimetic double-layer biofilm scaffold is prepared using natural small intestinal submucosa and chitosan, and Poly (lactic co glycolic acid) (PLGA) microspheres are used as controlled-release carriers of BMP-2, VEGF and PDGF. PLGA microspheres which could simulate the controlled release of BMP-2 in vivo and PLGA microspheres with sequential controlled release of VEGF and PDGF were combined into the inner and outer layers of the double-layer membrane scaffold respectively, and the bone marrow mesenchymal stem cells are inoculated in the periosteum The scaffold is used to construct a biomimetic periosteum with biological activity and loaded with stem cells, and then transplant it to the tendon-bone interface to promote tendon-bone healing. The biomimetic double-layer biofilm scaffold combines growth factors, repair scaffolds, and stem cells to reflect the combined application strategy. Therefore, this study will provide new ideas for the treatment of poor tendon bone healing after rotator cuff injury, and it is expected to reduce the incidence of rotator cuff nonunion and re-tear.This study also has the following limitations: due to the influence of intervention methods, this study cannot be strictly double-blind, which may affect the results; lack of quantitative evaluation indicators for rotator cuff healing and secondary arthroscopic observation; the current protocol has a short follow-up time. If necessary, we will plan to extend the follow-up to observe the medium and long-term effects.Data curation: Liang Ma, Yongtao Xu, Yongtao Xu.Funding acquisition: Yongtao Xu, Yongtao Xu.Investigation: Xiaolong Xu.Resources: Xiaolong Xu, Qin Pan.Software: Qin Pan.Supervision: Xiaolong Xu, Yongtao Xu.Writing – original draft: Liang Ma, Yongtao Xu, Yongtao Xu.Writing – review & editing: Liang Ma, Yongtao Xu, Yongtao Xu.


We report the first case of COVID-19 pneumonia in a preterm neonate in Mayotte, an overseas department of France. The newborn developed an acute respiratory distress by 14 days of life with bilateral ground glass opacities on a chest CT scan and a 6-week-long stay in the neonatal intensive care unit (NICU). This case report emphasizes the need for a cautious and close follow-up period for asymptomatic neonates born to mothers with COVID-19 infection. Vertical transmission cannot be excluded in this case. On 31 December 2019, the China Center for Disease Control and Prevention (CDC) reported an outbreak of pneumonia of unknown causes in the city of Wuhan [This pneumonia was linked to a novel coronavirus, the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), named coronavirus disease 2019 (COVID-19) [In Mayotte, one of the French overseas departments in the Indian Ocean with a population estimated at 256,000 and a significant birth rate of 10,000 births per year, a COVID-19 infection was first reported on 14 March 2020. As of 24 June 2020, the French public health agency (Santé Publique France) has identified 2467 cases of COVID-19 on the island . Little A 36-year-old multiparous woman, with a history of idiopathic bronchiectasis and gestational diabetes, was admitted to the delivery emergencies unit for active labor at 33 weeks of gestation. On admission, her body temperature was 37.4 °C, and her respiratory and hemodynamic parameters were within the normal ranges. Maternal antibiotic therapy and intramuscular corticosteroids for fetal pulmonary maturation were not administrated due to a rapid labor that lead to the vaginal birth, with meconium-stained amniotic fluid, of a preterm male infant. During the vaginal delivery, the mother developed a persistent cough and dyspnea, so she was suspected of being infected with COVID-19, and droplet and contact precautions were immediately initiated. RT-PCR of a nasopharyngeal swab from the mother was performed just after delivery and was positive for SARS-COV2  and her thoracic CT scan showed peripheral ground glass opacities associated with unilateral bronchiectasis. The neonate was born weighing 1830 g with an Apgar score of five at 1 min, seven at 5 min, and nine at 10 min. Skin-to-skin contact with his mother was not performed; he was transferred to the neonatal intensive care unit (NICU) on non-invasive intermittent positive pressure ventilation. He was in a closed incubator throughout his admission, and the mother’s access was not permitted in the NICU as she was symptomatic.On admission, he was clinically stable on non-invasive intermittent positive pressure ventilation with a mild transient respiratory distress that resolved within 24 h; his fractional concentrations of oxygen in inspired air (FiO2) remained at 21%. Furthermore, he was exclusively formula fed and was given prophylactic intravenous antibiotic therapy  that was stopped at 48 h as the blood culture was negative and the C-reactive protein (CRP) was normal. A nasopharyngeal swab RT-PCR was performed at 24 h of life and showed a positive result for SARS-COV2 with a viral load of 918,000 copies/uL. The RNA extraction was done with the QIAsymphony DSP Virus/pathogen Midi Kit (QIAGEN®). The chest X-ray showed normal lung aeration without pneumonia a. The infant remained stable and was switched to nasal continuous positive airway pressure at 2 days of life, and gradually weaned to low nasal flow cannula for 48 h; he was then completely weaned to room air at around 7 days of life.By 14 days of life, the infant developed a fever with progressive signs of increased breathing difficulties, such tachypnea and chest retractions, but his hemodynamic status remained normal.A thoracic CT scan revealed bilateral ground glass opacities and consolidations b. EchocaThe follow-up showed mild respiratory worsening by day 35; a secondary infection was ruled out and the patient was switched to nasal continuous positive airway pressure. He was weaned to room air after 45 days of life and discharged by day 50. Follow-up visits were planned for one week and one month following the discharge.Mothers with COVID-19 infection may present with obstetrical complications such as preterm labor, premature rupture of membranes, intrauterine growth restriction and neonatal death. In our case, preterm labor is presumably linked to the mother’s COVID-19 infection, as there was no other evident cause. In neonates, COVID-19 infection in the mother may cause severe acute respiratory distress and biological disorders, including thrombocytopenia and abnormal liver function . LaboratThe mode of transmission in our case is most likely to be vertical given the short time it took for the neonate to become SARS-CoV-2 positive after delivery (24 h), the absence of skin-to-skin contact at birth and throughout admission, and the absence of visits by the mother as she was symptomatic. In addition, the baby remained in a closed incubator with droplet and contact precautions, as COVID-19 was suspected during delivery; therefore, it seems unlikely that the infant was infected by the nosocomial spread of aerosolized virus or by an infected healthcare worker. Breastfeeding was excluded as the route of transmission as the infant was exclusively formula fed. However, our report has some limitations, including the fact that it presents a single case, a vaginal delivery and due to the absence of serology testing and PCR testing of the amniotic fluid, placenta and umbilical cord, as well as the mother’s vaginal secretions.Transmission through breastfeeding needs to be elucidated. In our unit, the use of maternal expressed breast milk is contraindicated in mothers with COVID-19. Skin-to-skin contact is not permitted given the benefit–risk balance. More studies are therefore needed in order to provide accurate evidence-based management guidelines. In the present paper, we noted a symptom-free interval between initial transient respiratory distress that was most likely related to the preterm birth, and the secondary respiratory distress that appeared by day 14, with a prolonged need for respiratory support and a longer length of stay compared to other reported pediatric studies ,13,14.This emphasizes the need for a cautious and close follow-up period for asymptomatic neonates born to mothers with COVID-19 infection who can develop severe pneumonia.Premature infants born to mothers with a COVID-19 infection may also have a COVID-19 infection, presumably via vertical transmission. Further studies are needed to confirm this route of transmission. The need for an extended and cautious follow-up period for asymptomatic neonates with COVID-19 should be kept in mind, as symptoms may appear secondarily with rapidly severe respiratory symptoms that may require prolonged respiratory support.


Soybean is a crop of major economic importance with low rates of genetic gains for grain yield compared to other field crops. A deeper understanding of the genetic architecture of yield components may enable better ways to tackle the breeding challenges. Key yield components include the total number of pods, nodes and the ratio pods per node. We evaluated the SoyNAM population, containing approximately 5600 lines from 40 biparental families that share a common parent, in 6 environments distributed across 3 years. The study indicates that the yield components under evaluation have low heritability, a reasonable amount of epistatic control, and partially oligogenic architecture: 18 quantitative trait loci were identified across the three yield components using multi-approach signal detection. Genetic correlation between yield and yield components was highly variable from family-to-family, ranging from -0.2 to 0.5. The genotype-by-environment correlation of yield components ranged from -0.1 to 0.4 within families. The number of pods can be utilized for indirect selection of yield. The selection of soybean for enhanced yield components can be successfully performed via genomic prediction, but the challenging data collections necessary to recalibrate models over time makes the introgression of QTL a potentially more feasible breeding strategy. The genomic prediction of yield components was relatively accurate across families, but less accurate predictions were obtained from within family predictions and predicting families not observed included in the calibration set. Soybean is a field crop of major importance due to its seed composition, containing approximately 40% protein and 20% oil. Its unique composition and scalable production make soy a key crop to world-wide food security , and a pThe first large-scale genetic assessment of complex traits was performed in the SoyNAM population, where 5600 genotypes from 40 bi-parental families sharing a common parent were phenotyped for various agronomic traits  was crossed to 40 founder parents that attempt to capture the diversity of public germplasm, each family comprising approximately 140 individuals. Among the 40 founder parents, 17 lines are U.S. elite public germplasm, 15 have diverse ancestry, and eight are planted introductions. The descriptions of parents are available -2. The phenotypes were collected in 10 field blocks, these being distributed as 4 adjacent blocks in 2013, 4 adjacent blocks in 2014 and 2 field blocks in different locations in 2015. In 2013 and 2014 the experiments were conducted at the ACRE farm, where each field block contained all 40 families with 35 recombinant inbred lines (RIL) per family, that is, one-quarter of the total number of RILs. In 2013 and 2014 RILs were not replicated, but the same checks were used across fields. In 2015, the experiments were conducted on 6 of the 40 SoyNAM families in two locations, ACRE and TPAC, with two replicates per location.The experiment was conducted under a modified augmented design, with a 7:1 lines-to-check ratio, in two Purdue University research centers: Throckmorton-Purdue Agricultural Center (TPAC) located in Throckmorton, Indiana, and at the Agronomy Center for Research and Education (ACRE) in West Lafayette, Indiana. The experiments were planted during the third week of May in two-row plots (2.9m × 0.76m), at a density of approximately 36 plants mThe number of pods and nodes was counted in the main stem, between phenological stages R5 and R7, averaging the counts of 3, 6 and 4 representative plants per plot in 2013, 2014 and 2015 respectively. The variable number of subsamples varied according to the resources available each year. The number of pods per node was obtained by the ratio. Grain yield was collected at harvest, converting the grain weight from individual plots to bushels per acre adjusted to 13% grain moisture. The number of days to maturity , as a random term of a mixed model. The mixed linear model was fitted with variance components based on restricted maximum likelihood (REML), computed using the R package lme4  or within trait between years (stability or genotype-by-environment correlation). The genetic correlations within-family were obtained as the Pearson’s correlation between the BLUPs of yield and yield components for pleiotropy analysis, as well as the correlations of yield components from year to year for stability analysis. We estimated the additive genetic correlation between yield and yield components for pleiotropy analysis, and yield components across years for stability analysis, for each of the 40 families using a multivariate GBLUP model. The GBLUP model was fit with REML variance components. For the multivariate polygenic analysis, we fitted the following multi-trait model:election  using itSince various signal detection strategies may capture different QTL . A briefThis method of an association study is based on the likelihood ratio between a model containing the marker of interest (full model) and a model without the marker (reduced model). Both models include a polygenic term that accounts for the population structure. The statistical model that describes this association study is tailored to NAM populations  within-family, (2) across-family, and (3) leave-family-out. Within- and across-family validations were performed as fivefold cross-validation, randomly selecting 80% of the data as a calibration set, and using the remaining 20% as a prediction target. The sampling and prediction procedure is repeated 25 times. Leave-family-out validation use 39 families to predict the family left out, and the procedure is performed to all 40 families. The prediction statistic is the predictive ability (PA), as the correlation between predicted and observed values.emCV of the R package bWGR , then load the Indiana dataset with the following command in R: data.All phenotypic and genotypic data are available in the R package SoyNAM available on CRAN. To access the data, install the SoyNAM package , which has been previously detecting in NAM families by et al. (2011) and The significant markers found from this study do not overlap with the signals found for grain yield . The reason why this phenomenon occurs is that different methods may improve how well the model detects the genetic architecture, but different types of cross-validation provide different information. Thus, gains associated to the choice of a prior are often considered negligible in comparison to increases in population size, better experimental practices, or more representative calibration sets (Note that the advantage provided by changing the model from GBLUP to BayesB is nowhere comparable to the difference in predictability between cross-validation methods (ion sets . As presion sets .When assessing genotype-by-environment, the total genetic correlation was larger the additive genetic correlation for all yield components , approxiFor the families with near-zero genotype-by-environment correlation, performing selections with a single year of data may not reflect into observable genetic gains in the coming years, and that collecting data from more environments may not necessarily increase the predictive ability of the yield components. Particularly for yield components, low genotype-by-environment correlations is not necessarily bad since the soybean yield plasticity relies on reallocating resources among yield components, which serves as a physiological response to mitigate yield losses under stress . For exaOur previous study (per se.From the standpoint of trait decomposition, the breaking down of grain yield into pods and nodes does not seem to be an effective approach since there is no strong evidence that these yield components are more heritable than yield  or stronIn our previous genomic prediction study (A major challenge of working with yield components is the data collection as the counting is highly subjective to human error, lowering the trait heritability and affecting the signal detection in downstream analysis. As deep learning methods for computer vision become increasingly population for phenotyping morphological traits (


Just as the human brain works in a Bayesian manner to minimize uncertainty regarding external stimuli, a deafferented brain due to hearing loss attempts to obtain or “fill in” the missing auditory information, resulting in auditory phantom percepts . Among various types of hearing loss, sudden sensorineural hearing loss (SSNHL) has been extensively reported to be associated with tinnitus. However, the reason that tinnitus develops selectively in some patients with SSNHL remains elusive, which led us to hypothesize that patients with SSNHL with tinnitus (SSNHL-T) and those without tinnitus (SSNHL-NT) may exhibit different cortical activity patterns. In the current study, we compared resting-state quantitative electroencephalography findings between 13 SSNHL-T and 13 SSNHL-NT subjects strictly matched for demographic characteristics and hearing thresholds. By performing whole-brain source localization analysis complemented by functional connectivity analysis, we aimed to determine the as-yet-unidentified cortical oscillatory signatures that may reveal potential prerequisites for the perception of tinnitus in patients with SSNHL. Compared with the SSNHL-NT group, the SSNHL-T group showed significantly higher cortical activity in Bayesian inferential network areas such as the frontopolar cortex, orbitofrontal cortex (OFC), and pregenual anterior cingulate cortex (pgACC) for the beta 3 and gamma frequency bands. This suggests that tinnitus develops in a brain with sudden auditory deafferentation only if the Bayesian inferential network updates the missing auditory information and the pgACC-based top-down gatekeeper system is actively involved. Additionally, significantly increased connectivity between the OFC and precuneus for the gamma frequency band was observed in the SSNHL-T group, further suggesting that tinnitus derived from Bayesian inference may be linked to the default mode network so that tinnitus is regarded as normal. Taken together, our preliminary results suggest a possible mechanism for the selective development of tinnitus in patients with SSNHL. Also, these areas could serve as the potential targets of neuromodulatory approaches to preventing the development or prolonged perception of tinnitus in subjects with SSNHL. Non-pulsatile tinnitus is a common otologic symptom characterized by conscious auditory perception in the absence of an external stimulus. This is often called a “phantom sound” because there is no corresponding genuine physical source of the sound , a complex and challenging emergency in the otology field, is typically defined as a sensorineural hearing loss of more than 30 dB across three consecutive frequencies in a pure-tone audiogram occurring within a 72-h period. Importantly, tinnitus was reportedly accompanied by SSNHL in 66–93% of cases  findings between SSNHL patients with and others without tinnitus (SSNHL-T and SSNHL-NT). Using whole-brain source localization analysis complemented by functional connectivity analysis, we aimed to determine the as-yet-unidentified cortical oscillatory signatures that could reveal the prerequisites for tinnitus development and to discuss the possible mechanism of the selective development of tinnitus in patients with SSNHL. Although this study includes a relatively small number of patients, which may have weakened the clinical implications of the results and statistical power, the results presented herein seem a more significant undertaking than we initially envisioned. Indeed, there is currently no consensus on the neurobiological markers for selective development of tinnitus in patients with SSNHL. Overall, our study stands out in this precision medicine era for incorporating neuroimaging in a tinnitus study to establish a future guide for the treatment of tinnitus in patients with SSNHL that incorporates neuroimaging as the “new normal.”We performed a retrospective review of the medical records of patients with unilateral idiopathic SSNHL who visited the outpatient clinic at Seoul National University Bundang Hospital (SNUBH) between January 2014 and March 2020. For the SSNHL-NT group, we were able to identify only 18 patients who met the criteria for unilateral SSNHL with no complaint of tinnitus. Two of the 18 were excluded due to an insufficient follow-up period . Of the remaining 16 patients, 3 were disqualified due to delayed emergence of tinnitus or significant hearing improvement during the follow-up period of at least 2 months after the onset of SSNHL. Ultimately, 13 patients were enrolled in the SSNHL-NT group. No patients in this group were diagnosed with Meniere's disease, vestibular schwannoma, or psychiatric/neurological disorders.As outlined in The hearing thresholds for seven different octave frequencies  were evaluated using pure-tone audiometry in a soundproof booth. The mean hearing threshold was calculated using the average of the hearing thresholds at 0.5, 1, 2, and 4 kHz . The impedances of all electrodes were maintained below 5 kΩ. Data were obtained at a sampling rate of 1,024 Hz and filtered using a high-pass filter with a cutoff of 0.15 Hz and a low-pass filter with a cutoff of 200 Hz. After initial data acquisition, the raw data were resampled at 128 Hz and band-pass filtered using a fast Fourier transform filter with a Hanning window at 2–44 Hz. After importing the data into Eureka! Software   without assuming a predefined number of active sources. The sLORETA solution space consists of 6,239 voxels (voxel size: 5 × 5 × 5 mm) and is restricted to the cortical gray matter and hippocampus, as defined by the digitized Montreal Neurological Institute (MNI) 152 template  was employed to estimate the scalp-recorded electrical activity in each of the eight frequency bands . The sLORETA software includes a toolbox for the functional localization of standardized current densities based on electrophysiological and neuroanatomical constraints  and known to relate to tinnitus according to previously published literature . Although the between-groups t-statistic was used for source localization and the paired t-test was used for connectivity analysis, these are nonparametric analyses based on 5,000 permutations. All analyses were done and illustrated using the R statistical package . All statistical tests were two-tailed, and P < 0.05 was considered significant.Statistical non-parametric mapping (SnPM) was adopted for permutation tests for source localization and functional connectivity. To identify between-group differences in resting-state cortical oscillatory activities, sLORETA built-in voxel-wise randomization tests  were used to perform nonparametric statistical analyses of functional images with a threshold The demographic and clinical characteristics of the two groups are summarized in P < 0.05) , the orbitofrontal cortex , and the pregenual anterior cingulate cortex  for the beta 3 and gamma frequency bands ( < 0.05) . No signP < 0.05)  for the gamma frequency band ( < 0.05) . For theThis is the first study to explore cortical activity and connectivity differences between SSNHL subjects with and those without tinnitus and to attempt to reveal the cortical oscillatory signatures for selective development of tinnitus among patients with SSNHL. In this study, the SSNHL-T group had abnormally increased activity in the FPC, OFC, and pgACC for the beta 3 and gamma frequency bands compared with the SSNHL-NT group. These findings suggest that auditory phantom percepts may develop when the brain experiences sudden decreased peripheral auditory input as the Bayesian inferential network updates the missing auditory information with the involvement of the pgACC-based top-down gatekeeper system. Furthermore, the lagged linear connectivity between the left OFC and the right precuneus was significantly increased for the gamma frequency band in the SSNHL-T group compared with the SSNHL-NT group, indicating that tinnitus deriving from Bayesian updating seems to involve the default mode network (DMN); thus, tinnitus seemed to be perceived as normal by the SSNHL-T group.The Bayesian brain model, an extension of a predictive brain model, has been suggested as an explanation for the development of tinnitus. According to this model, tinnitus is a response to peripheral auditory deafferentation that aims to reduce perceptual uncertainty , probably due to reduced canceling-out of irrelevant auditory input. Furthermore, increased activity in the parahippocampus and the pgACC for the theta and gamma frequency bands, as well as decreased activity in the auditory cortex, is found exclusively in tinnitus patients with hearing loss compared with those who have hearing loss but without tinnitus  deafferentation as well as with a dysfunctional top-down (descending) noise-canceling mechanism . The patients/participants provided their written informed consent to participate in this study.S-YL and J-JS led the analysis and interpretation of the results and drafted the first manuscript. BC, J-WK, and DD conceived the investigation and revised the manuscript for important intellectual content. All authors contributed to all aspects of the investigation, including methodological design, data collection and analysis, interpretation of the results, revision of the manuscript for important intellectual content, and approved the final version of the manuscript and agree to be accountable for all aspects of the work.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In this cluster-randomised controlled study (CoV-Surv Study), four different “active” SARS-CoV-2 testing strategies for general population surveillance are evaluated for their effectiveness in determining and predicting the prevalence of SARS-CoV-2 infections in a given population. In addition, the costs and cost-effectiveness of the four surveillance strategies will be assessed. Further, this trial is supplemented by a qualitative component to determine the acceptability of each strategy. Findings will inform the choice of the most effective, acceptable and affordable strategy for SARS-CoV-2 surveillance, with the most effective and cost-effective strategy becoming part of the local public health department’s current routine health surveillance activities. Investigating its everyday performance will allow us to examine the strategy’s applicability to real time prevalence prediction and the usefulness of the resulting information for local policy makers to implement countermeasures that effectively prevent future nationwide lockdowns.The authors would like to emphasize the importance and relevance of this study and its expected findings in the context of population-based disease surveillance, especially in respect to the current SARS-CoV-2 pandemic. In Germany, but also in many other countries, COVID-19 surveillance has so far largely relied on passive surveillance strategies that identify individuals with clinical symptoms, monitor those cases who then tested positive for the virus, followed by tracing of individuals in close contact to those positive cases.To achieve higher effectiveness in population surveillance and to reliably predict the course of an outbreak, screening and monitoring of infected individuals without major symptoms (about 40% of the population) will be necessary. While current testing capacities are also used to identify such asymptomatic cases, this rather passive approach is not suitable in generating reliable population-based estimates of the prevalence of asymptomatic carriers to allow any dependable predictions on the course of the pandemic.To better control and manage the SARS-CoV-2 pandemic, current strategies therefore need to be complemented by an active surveillance of the wider population, i.e. routinely conducted testing and monitoring activities to identify and isolate infected individuals regardless of their clinical symptoms. Such active surveillance strategies will enable more effective prevention of the spread of the virus as they can generate more precise population-based parameters during a pandemic. This essential information will be required in order to determine the best strategic and targeted short-term countermeasures to limit infection spread locally.This trial implements a cluster-randomised, two-factorial controlled, prospective, interventional, single-blinded design with four study arms, each representing a different SARS-CoV-2 testing and surveillance strategy.Eligible are individuals age 7 years or older living in Germany’s Rhein-Neckar Region who consent to provide a saliva sample  after completion of a brief questionnaire (two arms only). For the qualitative component, different samples of study participants and non-participants  will be identified for additional interviews. For these interviews, only individuals age 18 years or older are eligible.Of the four surveillance strategies to be assessed and compared, Strategy A1 is considered the gold standard for prevalence estimation and used to determine bias in other arms. To determine the cost-effectiveness, each strategy is compared to status quo, defined as the currently practiced passive surveillance approach.Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP).Strategy A1: Individuals (one per household) receive information and study material by mail with instructions on how to produce a saliva sample and how to return the sample by mail. Once received by the laboratory, the sample is tested for SARS-CoV-2 using Strategy A2: Individuals (one per household) receive information and study material by mail with instructions on how to produce their own as well as saliva samples from each household member and how to return these samples by mail. Once received by the laboratory, the samples are tested for SARS-CoV-2 using RT-LAMP.Strategy B1: Individuals (one per household) receive information by mail on how to complete a brief pre-screening questionnaire which asks about COVID-19 related clinical symptoms and risk exposures. Only individuals whose pre-screening score crosses a defined threshold, will then receive additional study material by mail with instructions on how to produce a saliva sample and how to return the sample by mail. Once received by the laboratory, the saliva sample is tested for SARS-CoV-2 using RT-LAMP.Strategy B2: Individuals (one per household) receive information by mail on how to complete a brief pre-screening questionnaire which asks about COVID-19 related clinical symptoms. Only individuals whose pre-screening score crosses a defined threshold, will then receive additional study material by mail with instructions how to produce their own as well as saliva samples from each household member and how to return these samples by mail. Once received by the laboratory, the samples are tested for SARS-CoV-2 using RT-LAMP.In each strategy, RT-LAMP positive samples are additionally analyzed with qPCR in order to minimize the number of false positives.The identification of the one best strategy will be determined by a set of parameters. Primary outcomes include costs per correctly screened person, costs per positive case, positive detection rate, and precision of positive detection rate. Secondary outcomes include participation rate, costs per asymptomatic case, prevalence estimates, number of asymptomatic cases per study arm, ratio of symptomatic to asymptomatic cases per study arm, participant satisfaction.Additional study components  include cost effectiveness of each of the four surveillance strategies compared to passive monitoring (i.e. status quo), development of a prognostic model to predict hospital utilization caused by SARS-CoV-2, time from test shipment to test application and time from test shipment to test result, and perception and preferences of the persons to be tested with regard to test strategies.Samples are drawn in three batches of three continuous weeks. Randomisation follows a two-stage process. First, a total of 220 sampling points have been allocated to the three different batches. To obtain an integer solution, the Cox-algorithm for controlled rounding has been used. Afterwards, sample points have been drawn separately per batch, following a probability proportional to size (PPS) random sample. Second, for each cluster the same number of residential addresses is randomly sampled from the municipal registries . The 28,125 addresses drawn per municipality are then randomly allocated to the four study arms A1, A2, B1, and B2 in the ratio 5 to 2.5 to 14 to 7 based on the expected response rates in each arm and the sensitivity and specificity of the pre-screening tool as applied in strategy B1 and B2. Based on the assumptions, this allocation should yield 2500 saliva samples in each strategy. Although a municipality can be sampled by multiple batches and the overall number of addresses per municipality might vary, the number of addresses contacted in each arm is kept constant.The design is single-blinded, meaning the staff conducting the SARS-CoV-2 tests are unaware of the study arm assignment of each single participant and test sample.Total sample size for the trial is 10,000 saliva samples equally allocated to the four study arms .For the qualitative component, up to 60 in-depth interviews will be conducted with about 30 study participants (up to 15 in each arm A and B) and 30 participation refusers (up to 15 in each arm A and B) purposefully selected from the quantitative study sample to represent a variety of gender and ages to explore experiences with admission or rejection of study participation. Up to 25 asymptomatic SARS-CoV-2 positive study participants will be purposefully selected to explore the way in which asymptomatic men and women diagnosed with SARS-CoV-2 give meaning to their diagnosis and to the dialectic between feeling concurrently healthy and yet also being at risk for transmitting COVID-19. In addition, 100 randomly selected study participants will be included to explore participants’ perspective on testing processes and implementation.Final protocol version is “Surveillance_Studienprotokoll_03Nov2020_v1_2” from November 3, 2020. Recruitment started November 18, 2020 and is expected to end by or before December 31, 2020.https://www.drks.de/drks_web/navigate.do?navigationId=trial. HTML&TRIAL_ID=DRKS00023271). Retrospectively registered 30 November 2020.The trial is currently being registered with the German Clinical Trials Register (Deutsches Register Klinischer Studien), DRKS00023271 (The full protocol is attached as an additional file, accessible from the Trials website (Additional file  Additional file 1.


Subcutaneous implantable cardioverter-defibrillators (ICDs) (S-ICDs) are advantageous because they eliminate the need for transvenous leads. However, just like in the case of traditional ICDs, inappropriate shocks are an unwanted complication that may result following their placement. In this case, we discuss the mechanism of an inappropriate shock in a patient with an S-ICD. This case highlights a limitation of long-term S-ICD sensing.Subcutaneous implantable cardioverter-defibrillators (ICDs) (S-ICDs) are considered by many to be advantageous over more traditional ICDs due to their elimination of the need for transvenous leads. However, with either type of device, inappropriate shock can occur as an unwanted complication. The Boston Scientific Postmarket S-ICD Registry (EFFORTLESS) noted that 8.3% of its 581 patients with S-ICDs had inappropriate shocks (average of 2.2 shocks per patient). This translated to 48 patients over a period of 21 months ± 13 months, with the registry beginning in 2009.A 41-year-old female with nonischemic cardiomyopathy received a model 1010 SQ-RX S-ICD . Adequate QRS sensing was present at the time of implantation and during ventricular fibrillation (VF) induction. The secondary sensing configuration was used and VF therapy was programmed to 230 beats per minute (bpm) with a conditional zone of 200 bpm.. The patient stated that she had frequently used her massager without issue prior to this episode.Three-and-a-half years after implantation, the patient experienced an ICD shock while using a TENS unit on her neck, axilla, and back . Additional low-amplitude, high-frequency signals with amplitudes of 0.35 mV to 0.5 mV were noted during the event and sensed by the patient’s S-ICD as intrinsic cardiac electrical activity. As the heart rate perceived by this patient’s device was consistent with VF, an ICD shock was administered. Closer examination of the event clearly demonstrated that the patient’s native QRS  was present during, and independent of, these high-frequency signals, thereby confirming the presence of noise.A review of the electrograms demonstrated a low-amplitude QRS  that was temporally related to the use of the modulated TENS device. The patient was advised not to use the TENS device again. The QRS noted at implant was 1.6 mV  and the true QRS at the time of the shock episode was 0.8 mV . Although her QRS was now lower than at the time of implantation, as sensing was stable and similar to other vectors, we did not make additional device changes.External analysis of this event demonstrated a single dominant frequency consistent with electromagnetic interference (EMI) We speculate that the combination of a low-amplitude native QRS with external noise resulted in inappropriate VF detection and delivery of therapy. The progressive decrease in our patient’s QRS voltage with time may possibly explain the timing of her inappropriate shock, which occurred for the first time at 3.5 years after S-ICD implantation, despite ongoing use of the same TENS device since the time of her S-ICD implant procedure.EMI is an electrical signal generated by an internal or external source that falls within the sensing frequency spectrum of an implantable cardiac electronic device that may result in, or trigger, inappropriate device behavior. Although sensing algorithms and low-pass filters within implantable devices are designed to mitigate inappropriate device behaviors, they are not infallible. One may speculate that the sensing vectors in the S-ICD created by the superficial location and wide antennae of the device can increase its susceptibility to EMI and myopotential oversensing as compared with transvenous ICDs. In our case, the low sensed QRS amplitude may have contributed to the inability to filter out the external noise, as the estimated starting peak for the refractory period would be reduced, allowing for a lower or more sensitive absolute floor for detection.2 In the former situation, one may simply instruct patients on the need to avoid the device causing the EMI, whereas the latter may require additional device reprogramming or repositioning.Offline spectral analysis of the event was invaluable, as it confirmed EMI rather than myopotential oversensing. The latter is another rare cause of inappropriate therapy deliverance in S-ICD patients.6 However, to our knowledge only one report of an inappropriate shock due to TENS use has been reported before in an S-ICD patient.7 Our case highlights the potential impact of long-term changes in device sensing on the delivery of ICD therapy in special situations, such as the presence of EMI. This case should remind clinicians of the limitations of ICD sensing and promote patient education on day-to-day activities that may adversely impact their ICDs.While the use of TENS is contraindicated in patients with ICDs, patients may still utilize this therapy despite recommendations. Indeed, our case is not unique, as there are prior reports of inappropriate shocks due to TENS use in transvenous ICD patients.


Our understanding of all aspects of coronavirus-19 (COVID-19) is evolving. Some aspects already appear clear: children are significantly less likely to be diagnosed with SARS-CoV-2 than adults . NeverthDetails about SARS-CoV-2 infection in children first emerged in small, retrospective case series from China. Children in these series form a relatively heterogenous population, with up to 40% being asymptomatic. These early studies identified that children who displayed symptoms were less likely to have a cough and fever, but more likely to have vomiting or diarrhoea compared to adults and were also likely to present to hospital earlier in the disease. These findings are borne out in larger case series from China, Spain and the USA where fever is reported in 40–60% of children, cough in 19–65% of children and diarrhoea and vomiting in approximately 10% of children diagnosed with SARS-CoV-2 infection when the number of asymptomatic carriers included in the study are low .Predictive markers of severe SARS-CoV-2 infection are not yet fully understood, with ongoing evaluation of the effects of gender, ethnicity, age, co-morbidities and biochemical and radiographic features on outcome. Much of this evaluation will be undertaken in adults, but its applicability to children, particularly younger children and infants, has questionable relevance. In the published literature, there do not appear to be specific co-morbidities which increase the risk of requiring hospitalisation for SARS-CoV2 infection. A small Chinese study describes 46% of hospitalised children having cardiac co-morbidities, none of whom required intensive care support and were successfully discharged. Two children with underlying congenital heart disease required mechanical ventilation and one of these children also required haemofiltration. The USA reports that 7% of children diagnosed with COVID-19 and with details entered into the CDC database have cardiovascular disease and up to 12% having chronic lung disease, including asthma [There is good evidence in adult patients that lymphopaenia is a good predictor of disease severity for COVID-19. The paediatric studies which report blood results have small numbers of patients with a variety of phenotypes within them , making it difficult to draw conclusions about the findings . HoweverComputed tomography (CT) has been used extensively in China as a diagnostic tool for SARS-CoV-2. Reverse transcriptase polymerase chain reaction (RT-PCR) has been reported as having a false negative rate up to 30% , promptiImportantly, the article highlights CT abnormalities which can be found in the majority of children with confirmed COVID-19. Ground-glass opacity, local and bilateral patchy shadowing are the three most common findings and are reported in 36–100% of children with confirmed SARS-CoV-2 in other series , 6. CT aThe need for admission to the intensive care unit (ICU) and the mortality rate of children with COVID-19 are unclear. At least 7/246 (3%) Chinese children have been reported as requiring intensive support with 1 death reported . The ratMinimal data are published on the children who are affected by severe SARS-CoV-2. These small numbers make it difficult to draw conclusions about which children are at risk from severe complications of SARS-CoV-2 infection and death. However, the overall message about COVID-19 in children is reassuring. Infection rates of SARS-CoV-2 in children are lower than those in the general population. The majority of infected children are asymptomatic or have mild disease, and do not require invasive support. There is a suggestion that children with cardiac co-morbidities may be at higher risk of developing severe COVID-19. This is based on a limited number of small case series and incomplete reporting into CDC database which may reveal a reporting bias.The presenting features of COVID-19 differ in children from adults, with lower rates of fever and cough and more prominence of vomiting and diarrhoea. Currently, haematological, biochemical and radiological findings in children infected with COVID-19 are not diagnostic and do not consistently predict severe disease.Prospective research is required to help identify any patient groups who are at risk of developing severe infection and features which may be used to identify these children early in the disease course.


Glycosylation is one of the most crucial posttranslational modifications of proteins, containing a remarkable amount of biological information. The alteration of glycosylation is closely associated with certain diseases. Exploring glyco-code in the development of diseases is a hot topic in recent years. Esophageal squamous cell carcinoma (ESCC) is the primary pathological histology in developing countries and a severe threat to human health. Although the glycan profiles in the blood samples of ESCC patients were analyzed using glycomic and glycoproteomic methods, the difference of salivary glycopatterns between healthy subjects and ESCC patients is not explicit yet. In the present study, ESCC patients (n = 16) and healthy volunteers  were enrolled. The glycomic strategy combining lectin microarray and lectin blotting was employed to investigate and confirm the altered salivary glycopatterns. Datura stramonium (DSA) was selected to isolate the GlcNAc or Galβ1-4GlcNA-containing glycoproteins due to the distinct difference between ESCC patients and HVs. The N-glycans from DSA-enriched glycoproteins were released by PNGase F and further identified by MALDI-TOF/TOF-MS to obtain the precise structural information of the altered glycans. As a result, the glycopatterns recognized by 13 lectins  showed significant alterations in ESCC patients’ saliva. The ESCC patients showed higher levels of GalNAc and Gal, sialic acid, and GlcNAc expression profiles and lower levels of mannose and fucose expression profiles. The MALDI-TOF/TOF-MS results indicated that the proportion of the GlcNAc or Galβ1-4GlcNAc-containing N-glycans was increased in ESCC patients (79.04%) compared with HV (63.20%), which was consistent with the results of lectin microarrays. Our findings provide comprehensive information to understand the complex physiological changes in ESCC patients. And the altered salivary glycopatterns such as GlcNAc or Galβ1-4GlcNAc-containing N-glycans recognized by DSA might serve as potential biomarkers for the diagnosis of ESCC patients. Esophageal cancer (EC) ranks as the seventh most prevalent cancer with more than 570 thousand new cases and the sixth most lethal cancer with over 500 thousand deaths worldwide . EC is aThe assembly of glycans is complex enzymatic progress catalyzed by a series of glycosyltransferases and glycosidases, which starts in the endoplasmic reticulum and matures in the Golgi apparatus . As the In this study, the altered salivary glycopatterns related to ESCC were investigated and confirmed by the integrated glycomics strategy. To acquire precise structural information of the altered glycans between ESCC and healthy volunteers (HV) cases, the glycoproteins were isolated by lectin-mediated affinity capture, and the N-glycans were released and purified from the isolated glycoproteins and then identified using MALDI-TOF/TOF-MS. The purpose of this study is to clarify the altered salivary protein glycopatterns related to ESCC and to identify the precise structures of the altered salivary glycopatterns, which may help us to understand the complex physiological changes in ESCC patients.The collection and use of the whole saliva for research in this study were approved by the Ethical Committee of Northwest University  and the Second Affiliated Hospital of Zhengzhou University . Written informed consent was received from participants. And this study was conducted by the ethical guidelines of the Declaration of Helsinki. After a standardized endoscopic procedure and histopathological evaluation, the individuals diagnosed with primary ESCC (n =16) were enrolled in this study. The age- and sex-matched HVs (n = 25) were recruited from the health checkup center in the same hospital where they underwent a medical examination. All the participants had no significant differences between the two groups regarding demographic, socioeconomic, and lifestyle characteristics, and those who received preoperative radiotherapy, chemotherapy, chemoradiotherapy, or antibiotic therapy were excluded from this study. The clinical characteristics of HVs and ESCC patients were summarized in The collection protocol has been described in previous literature . The colhttp://acgt.cs.tau.ac.il/expander/) and Multivariate Statistical Package (United Kingdom), respectively. And the p value was derived from the nonparametric Mann–Whitney test using the GraphPad Prism software .The lectin microarrays were produced using 37 lectins with different binding preferences covering N- and O-linked glycans . The Cy3The expression levels of glycan structures were analyzed by lectin blotting as described previously . The poo4O7, 180 mM H3BO4, 150 mM Na+, pH 7.4), respectively, and reacted with DSA solution (DSA dissolve in coupling buffer) according to the protocol (The epoxy-coated magnetic particles (2 mg) were rinsed with ethanol and coupling buffer  and incubated with the DSA conjugates . After 3 h gentle shaking, the conjugates were washed using washing buffer  to remove unbound proteins, and the glycoproteins bound to the conjugates were eluted with an eluting buffer .The GlcNAc and Galβ1-4GlcNAc-containing glycoproteins were isolated from ESCC patients and HVs using DSA-magnetic particle conjugates as described previously . Briefly4HCO3 buffer and incubated with trypsin . The trypsin in the mixture was inactivated by heating  and then the PNGase F  was added to release the N-glycans . Subsequently, the digest was subjected to HyperSep Hypercarb SPE cartridges  to remove peptides. The purified N-glycans were collected and lyophilized.The isolation and purification of N-glycans were performed based on previously described methods . The glyThe N-glycans were characterized by matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectroscopy  as described previously . GlycansA total of 16 ESCC and 25 HV samples were detected by lectin microarrays independently to investigate the altered salivary protein glycopatterns associated with ESCC patients. The layout of the lectin microarrays and typical representative images were listed in To further investigate the alteration of salivary protein glycopatterns in ESCC patients, the Mann–Whitney test was used to compare the variance between ESCC and HV groups. In total, 13 lectins exhibited significantly altered NFIs in ESCC cases. As shown in SDS-PAGE and lectin blotting analysis were performed with silver staining and Cy5-labeled lectin staining to confirm the different abundances of glycopatterns in pooled saliva from ESCC and HV cases. The results of SDS-PAGE demonstrated that the distribution and abundance of salivary protein bands were similar in ESCC and HV cases . The resTo obtain the GlcNAc, Galβ1-4GlcNAc N-glycan structures of glycoproteins in saliva from ESCC and HV cases. The glycoproteins were isolated using the DSA-magnetic particle conjugates, then N- glycans were released by PNGase F, purified by HyperSep Hypercarb SPE cartridges, and identified by MALDI-TOF/TOF-MS, respectively. A total of 48 and 56 N-glycan peaks from the pooled saliva samples of ESCC and HV were identified and annotated with proposed structures in Despite the continuous improvement in medical technology and comprehensive treatment, the prognosis of ESCC patients remains poor . It is rDecades of research have demonstrated that aberrant protein glycosylation often occurs in the development of tumors, and it has been shown that specific tumor-associated glycans are expressed in the precursor lesions of different types of cancer, which makes them potentially powerful early diagnosis markers . GlycomiN-glycans was increased in ESCC (79.04%) compared with HV (63.20%), which was consistent with the results of lectin microarrays. Compared with our previously published data of the other important gastrointestinal cancer, we found that the glycopatterns of ESCC and GC are obviously different  revealed significant alterations of the salivary glycopatterns between ESCC and HV cases. As a result, ESCC patients showed higher levels of GalNAc and Gal expression profile, sialic acid expression profile, GlcNAc expression profile, and lower levels of mannose expression profile and fucose expression profile. The lectins of DSA, ECA, LCA, PSA, and UEA-1 were randomly selected to confirm the differentially expressed sugar patterns. The lectin of DSA has an affinity to both GlcNAc and Galβ1-4GlcNAc, which can be used as an effective tool for the analysis of complex-type N-glycans. We used the DSA-magnetic particle conjugates to isolate the glycoproteins. And the glycan profiles of DSA-isolated glycoproteins were analyzed by MALDI-TOF/TOF-MS after liberation and purification. The results indicated that the proportion of the GlcNAc or Galβ1-4GlcNAc-containing ifferent . The NFIifferent . These rOur analyses have some limitations. First, it is a pilot study, the samples recruited are not enough, and an independent validation group is absent in this study. Second, the derivatization of sialic acids, such as permethylation and ethyl esterification, is needed for more accurate measurements of the released N-glycan profile. But our study provided comprehensive information of saliva glycopatterns from ESCC and the resolution of ESCC-associated glycopatterns that may contribute to understanding the complex physiological changes of ESCC patients. And the altered salivary glycopatterns such as GlcNAc or Galβ1-4GlcNAc-containing N-glycans recognized by DSA might be served as potential biomarkers for the diagnosis of ESCC patients.


Considering the COVID-19 emerging and rapidly evolving situation associated with increased levels of mortality and infectivity risks, the detection and identification of new tests in a fast, safe, and accurate measures would have a high impact regarding prompt clinical and epidemiological management decisions. The combination of real-time polymerase chain reaction and the immunoglobulin class M–immunoglobulin class G antibody serology testing can be a powerful strategy for more accurate severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection diagnosis with less false results slipping through the cracks. The following viewpoint is describing the immunological response to SARS-Cov-2 infection and its implication in the selection of the appropriate diagnosis tools. In 2011, the world experienced global pandemic caused by Middle East Respiratory Syndrome coronavirus (MERS-CoV).1 The latest CoVs to spill over into the human population emerged in Wuhan City, Hubei Province, China, in December 2019. This virus was termed SARS-CoV-2 by the International Committee on Taxonomy of Viruses (ICTV) for SARS-CoV-2. The newly identified virus was responsible for the outbreak in Wuhan and patients with confirmed infection had typical symptoms of fever, cough, shortness of breath, and in severe cases, pneumonia.3 Worldwide situation spread rapidly, with >34.8 million confirmed cases reported globally and 1 million deaths as of October 4, 2020.4 Considering the emerging and rapidly evolving situation associated with increased levels of mortality and infectivity risks, the detection and identification of new tests in a fast, safe, and accurate measures would have a high impact regarding prompt clinical and epidemiological management decisions.5Coronaviruses (CoVs) belong to the Orthocoronavirinae's subfamily of the family Coronaviridae. It is a group of enveloped positive-sensed single-stranded RNA viruses. The emerging CoVs have caused recent pandemics of respiratory infectious diseases with high mortality.7 Even though real-time PCR has advantages, this technology has several limitations that warrant some attention in the context of COVID-19.8–11 First of all, it incurs a huge cost of instrumentations and consumables, and requires certified laboratories and specifically trained personnel. These tests have limited standardized protocols, and are laborious in operation and time-consuming. Another consideration concerns reported false negatives for real-time PCR of SARS-CoV-2.12–14 Increased risk of false negative in pathogen detection, particularly for new emerging or highly variable pathogens is one of major limitations. Any low abundance target will be more likely to experience the Monte Carlo effect because the probability of primer annealing is lower.16 Because of this, real-time PCR appears weak for accurate data interpretation and is inadequate for rapid and simple diagnosis. A cautionary warning has already been issued for multiple routes transmission and needed strategy to accurately predict infection status.17The standard method of suspected infection diagnosis is real-time polymerase chain reaction (PCR), based on viral spike genes, for detection of SARS-CoV-2 nucleic acid in sputum, throat swabs, and secretions of the lower respiratory tract samples.17 It has been shown that infected patients can harbor the virus in the intestine at the early or late stage of disease progression. More interestingly, it has been noted that cases with positive blood viremia had negative swabs. Furthermore, the diagnosis of SARS-CoV-2 is confirmed with lower respiratory tract specimens and no upper respiratory tract swabs are investigated for the viral RNA detection, which may conclude bias results during the early infection stage and explain high false negatives in nucleic acid PCR test.3 This strongly implies that these patients would likely be considered as SARS-CoV-2 negative through established routine surveillance practices and discharged only on the basis of negative oral swabs, putting them at a high risk of transmission.Current strategy for SARS-CoV-2 diagnosis is to detect viral RNA in oral swabs, which is not perfect and comes with challenges. The virus can be present in anal swabs or blood of patients, whereas oral swabs testing negative.18 The neutralizing antibody-mediated humoral response is very potent in neutralizing viral infectivity and prevents reinfection. Small-scale serological screening of SARS-CoV-2 with limited data was reported.7 It has been shown that, after the onset of the disease, specific immunoglobulin class M (IgM) reached peak titer in day 9 and immunoglobulin class G (IgG) levels peak within about 2 weeks. Furthermore, patients' serum showed cross-neutralizing activity in vitro eliciting a protective humoral immune response. IgG and IgM viral antibodies were identified in all patients regardless of swabs results.7 However, it remains to be investigated if kinetic and titer of specific antibody correlates with disease severity. This implied that serological assays can play an important role in SARS-CoV-2 case detection and surveillance. Serology are less likely to miss infected people and can help both indication of immune status and diagnosis of acute infection.For better diagnostic strategy, dissecting the dynamic of immune response to SARS-CoV-2 is essential. Using accumulated knowledge learned from the outbreak of SARS-CoV and MERS-CoV, one can anticipate the gap on human immune responses to SARS-CoV-2 infection.19 Authors highlighted that progressive increases in plasma SARS-CoV-2-binding IgM and IgG antibodies happen before the resolution of symptoms and clinical recovery. This immune response to SARS-CoV-2 was described as similar to what we observe in influenza. Using blood samples for diagnosis of SARS-CoV-2 seems to be the best-case scenario for rapid, simple, and highly sensitive detection method. It is well known that IgM antibodies are usually the first to arrive to an infection scene, before high affinity, and longer-lasting IgG responses generation. The detection of IgM antibodies indicates recent exposure to the virus and hence a possible presence in blood. IgM antibodies are often considered as the most sensitive indicator of acute infection. The presence of IgG antibodies generally indicates past exposure and immunity.20 Serological screening for both IgM and IgG throughout the time-course of infection could increase diagnostic accuracy, help monitoring course disease during and after treatment, and help identify reservoirs that may amplify outbreaks.12 The advantages of serological IgG–IgM combined antibody tests are that they are material and time-saving tools, suitable for testing large samples, require no specialized equipment or technique and can be performed by most hospital or clinic laboratories. Compared with real-time PCR, they are relatively inexpensive, simple to perform, and only requires minimal training.Shedding light on immune responses would be valuable for designing diagnosis and therapeutic intervention for COVID-19. The first contributions to mapping the breadth and kinetics of serological response to SARS-CoV-2 comes from Thevarajan et al.19 This promising research could also help us identifying earlier patients at risk of severe symptoms and which patients will develop milder cases, based on humoral and cellular immune parameters. It is unclear how the immune system fights SARS-CoV-2 and how it works for recovered patients. Research in this area could provide more insight into how postinfection immunity develops for SARS-CoV-2, and if immunization is temporary or longer-lasting to anticipate seasonal outbreaks.Gaining a deeper understanding of humoral and cellular immune parameters in larger cohorts of COVID-19 will help predicting disease courses, identifying new tools for diagnosis and disease severity management, and developing protective vaccines with maximum efficacy.11 Serological information may simplify and promote contact tracing and surveillance at the local, regional, national, and international levels. Serological immunity can also accelerate return-to-work outcome of frontline health workers, which will be potentially re-exposed to SARS-CoV-2. Antibody testing may be helpful in testing sensitivity of real-time PCR assays for detecting viral infection.As asymptomatic carriers could spread SARS-CoV-2 virus and make the current outbreak control more difficult, another promising application of serological tests is screening asymptomatic carriers, which is vital to curb the epidemic.In summary, in the face of a rapidly changing and never-before experienced situation, and unfortunately in the absence of any effective therapeutics or vaccines, we have to leverage the power of testing to overcome the pandemic confronting us. One must be aware that we are facing a new virus and uncharted territory, exposing us to new questions and challenges we had yet to be ready to ponder on: what is the most appropriate test, and for whom and when? What to test? How often to test? And, what to do with test results?Today, in the thick of the outbreak, the need for a rapid, simple to use, sensitive, and accurate diagnostic test is critical to quickly identify symptomatic and asymptomatic cases of SARS-CoV-2, reduce virus shedding and transmission, and ensure timely treatment of patients. The combination of real-time PCR and the IgM–IgG antibody serology testing can be a powerful strategy for more accurate SARS-CoV-2 infection diagnosis with less false results slipping through the cracks.


I2 = 72%), and no statistically significant differences for OS , DFS , and DSS . The high degree of heterogeneity of the studies is a notable limitation. Conservative management and appropriate follow-up instead of bilateral surgery would be justifiable in selected patients. These findings highlight the importance of shared-decision making in the management of patients with small, low-risk DTC.Recently, the management of patients with low-risk differentiated non-medullary thyroid cancer (DTC), including papillary and follicular thyroid carcinoma subtypes, has been critically appraised, questioning whether these patients might be overtreated without a clear clinical benefit. The American Thyroid Association (ATA) guideline suggests that thyroid lobectomy (TL) could be a safe alternative for total thyroidectomy (TT) in patients with DTC up to 4 cm limited to the thyroid, without metastases. We conducted a meta-analysis to assess the clinical outcomes in patients with low-risk DTC based on the extent of surgery. The risk ratio (RR) of recurrence rate, overall survival (OS), disease-free survival (DFS) and disease specific survival (DSS) were estimated. In total 16 studies with 175,430 patients met the inclusion criteria. Overall, low recurrence rates were observed for both TL and TT groups (7 vs. 7%, RR 1.10, 95% CI 0.61–1.96,  Differentiated non-medullary thyroid cancer (DTC) is the most common endocrine malignancy and accounts for 85% of thyroid cancer . EpidemiThere is controversy on the optimal treatment of patients with DTC, raising the question whether some of the patients with low-risk DTC might be overtreated and therefore at risk for complications of these treatments without having a clear benefit in terms of oncological outcome. Until recently, total thyroidectomy (TT) followed by postoperative radioactive iodine (RAI) treatment to ablate the thyroid remnant tissue or as an adjuvant therapy was recommended for the majority of patients by many international guidelines. Over the past decade, clinical practice has shifted towards a more individualized approach. In particular, better understanding of the biology and excellent outcomes of these tumors has led to recommending a more conservative approach for low-risk patients . TT is nThe role of conservative limited surgery versus more extensive surgery for patients with low-risk DTC has not been studied in prospective randomized controlled trials, as large sample size and a long-term follow-up would be required ,20. TherThe purpose of this meta-analysis is to compare the clinical outcomes in adult patients with low-risk DTC based on two surgical modes, TL versus TT, and discuss in that light the arguments for and against TL and TT in low-risk DTC according to the definition and recommendation of the ATA.The meta-analysis was performed in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-analysis (PRISMA) guidelines . We searTwo authors (A.B. and A.K.) extracted all relevant data independently. The following data were extracted: year of publication, first author’s name, country of study origin, % female, mean age, study design, surgical approach, follow-up duration, use of radioactive iodine (RAI), inclusion of high-risk patients and outcomes assessed. The data were reviewed by a third author (R.N.-M.). Disagreements were resolved through consensus.We used a component approach to assess risk of bias for all included studies. The components were subdivided into five separate domains based on the Cochrane Risk-of-Bias In Non-Randomized Studies-of Interventions (ROBINS-I tool) . The folDomain 1: Bias due to confounding (see below)Domain 2: Bias in selection of participants into the studyInclusion of patients .Definition of low risk thyroid cancer?Domain 3: Bias in classification of intervention.Were intervention groups clearly defined?Domain 4: Bias due to missing dataTNM-stage available (see below)Reporting of high-risk patients (see below)Reporting of RAI treatment (see below)Reporting number of patients included per treatment arm (see below)Domain 5: Bias in measurement of outcomesLoss to follow-up (<5% is considered low risk of bias)Criteria for extent of surgery (see below)Reporting of outcome definitionMatched cohorts reported no bias of confounding. As the type of surgery procedure was not always clearly described, adequately reporting the criteria used for the extent of surgery was considered a low risk of bias. Availability of TNM-stage was considered adequate if the article mentioned T1–T4. Reporting of high-risk patients was considered adequate if the article mentioned T3–T4, N1 and/or M1 stage or mentioned numbers of high-risk patients with well-defined criteria for high risk. Reporting of RAI treatment was considered adequate if the numbers of patients receiving RAI treatment were mentioned. Reporting the number of patients included per treatment arm was considered adequate if the article mentioned the number of patients for each type of surgery.2 and continuity correction of 0.5 in studies with zero cell frequencies). Heterogeneity among studies was evaluated using the I2 statistic [Pooled RR’s, 95% confidence intervals (95% CI) and 95% prediction intervals were calculated using random effects models . Studies were considered to have a low risk of bias if they met at least ten (out of eleven) of the above-mentioned criteria for low risk of bias. Articles were considered intermediate risk of bias if they met up to nine of the above-mentioned criteria for risk of bias. The potential publication bias was evaluated with a funnel plot .In total, 106 abstracts and titles were obtained through electronic search, and 17 articles identified through manual search. The detailed search yielded no randomized controlled trials. Forty-four articles were found eligible, of which the results of 23 articles were examined in detail; 16 studies met the inclusion/exclusion criteria, which resulted in 175,430 patients. I2 = 72%)  . No signI2 = 85%). There was a marked statistical heterogeneity . There was a marked statistical heterogeneity . There was a marked statistical heterogeneity  had a matched cohort and no bias of confounding. Definition of low-risk patients was adequately reported in 13 articles (81%). Inclusion of consecutive patients or use of a random sample was explicitly stated in 15 articles (94%). Loss to follow up was reported in six articles 37.5%), one of these six articles reported a loss to follow up of <5%. Four of these six articles also included patients who underwent less than a lobectomy or a subtotal thyroidectomy, procedures that are regarded as inferior in terms of oncologic outcome ,38,39,463% had a .5%, one One article showed low risk of bias (met ten out of eleven criteria), and five articles showed intermediate risk of bias (met nine out of eleven criteria). There was no study that met all eleven criteria. The risk of bias for each study is reported in Publication bias was examined by funnel plots. No clear indication of publication bias was observed for recurrence rates .To our knowledge, this study is the first meta-analysis focusing on the outcome of surgical extension in terms of recurrence and survival in patients with low-risk DTC 1–4 cm without clinical evidence of locoregional of distant metastases. Our meta-analysis shows no statistically significant differences between TL versus TT regarding DTC-related recurrence, DSS, OS and DFS rates.The effect of the extent of surgery for patients with low risk DTC on patients’ outcome has long been a matter of discussion partly because of a lack of evidence-based randomized data from prospective studies. The early study of Mazzaferri et al. showed a statistically significant advantage of TT in comparison to TL regarding both recurrence and survival . These rLower complication rate, avoiding iatrogenic hypothyroidism and shorter duration of surgery are important arguments for preferring TL. Nonetheless, there could be several reasons for preferring a TT. One reason is the concern regarding an increased risk for recurrent or residual disease . In addition, distant metastases could be missed and might only become apparent on whole body scans performed after ablative treatment with RAI, as limited surgery is usually not followed by treatment with RAI. In this meta-analysis, both local and distant metastases recurrence rates were low. There was no statistically significant difference of local recurrences in TL compared to TT, and distant metastases recurrence rates were higher in TT. Of note, only five studies ,29,36,45A second reason for preferring TT is that bilateral tumor localizations are found in 20% to 85% of patients with PTC after total or completion thyroidectomy, but the majority are considered occult carcinoma ,53,54. PA third reason is that microscopical lymph node metastases are also likely to be found in histopathological samples  , but theA fourth and important reason to prefer TT over TL is routine use of RAI ablation in the majority of patients with tumors 1 cm or greater. In terms of outcome, RAI ablation has not been shown to clearly benefit the low-risk group patients in the absence of any high-risk features, while being associated with some side effects ,57,58. HOur study has some limitations which should be considered by the interpretation of the data. Due to the lack of randomized prospective studies, our estimates were based on non-randomized retrospective studies which are prone to selection bias, are limited by the availability of patient data and are characterized by a high degree of heterogeneity. The sensitivity analysis demonstrates that there is one individual study with an extreme effect on the overall RR for recurrence rates. Hassanain et al. showed a much higher recurrence rate in the more extensive surgery patients, which indicates selection bias . Only twWhen selecting the optimal surgical extent in patients with low-risk DTC, the goal is to minimize the risk of death from disease and recurrence, while avoiding potential iatrogenic injury from overtreatment. Disease-related characteristics are critical, and risk stratification is paramount. Individual patient values and preferences, as well as surgeons’ experience and quality of follow-up and patients’ adherence to follow-up should also be considered when making this decision .TL is a safe procedure for treatment of low-risk DTC patients, with excellent oncological outcomes and potentially lower treatment side effects. Initial risk assessment and ongoing dynamic risk stratification can reduce the uncertainty about disease persistence and individualize follow-up and treatment in order to reduce recurrence rate, the risk of completion thyroidectomy, and patient anxiety. However, it is important to inform the patients about the expected outcomes for both TL and TT with respect to morbidity, need for hormone replacement and the risk of reoperation. Thus, shared decision-making should be considered when the choice for the most appropriate surgical approach is being made.


Soto et al is quite interesting1. SARS-CoV2 (COVID19) is a novel coronavirus that has caused a critical public threat worldwide. How to prevent the COVID-19 outbreak is a crucial issue and community-based preparedness is important. SARS-CoV2 is transmitted through droplets and contact transmission routes, and the Taiwan government has taken many proactive and aggressive actions to prevent the transmission of COVID-19, including wearing face masks. These strategies have not only controlled COVID-19 transmission but also reduced influenza infection in Taiwan.The study of face masks use in Latin America reported by Although Taiwan is geographically close to China, the number of confirmed cases with COVID-19 in Taiwan is significantly lower than in other neighboring countries, with fewer than 450 confirmed COVID-19 cases and limited community transmission. The Taiwan government has acted swiftly since January to prevent the COVID-19 outbreak crisis by implementing measures such as border control, case identification and isolation, quarantine of suspected cases, extending holidays, travel restriction, education and big data integration. In addition, the export of face masks was prohibited on January 24, 2020 to avoid a shortage. In addition to these proactive early decisions, the Taiwan government acquired all available face masks and assigned most of them to hospital staff first in order to protect them from COVID-19. Moreover, mask making machine manufacturers were asked to step up face mask production capacity, which rose from 1.3 million per day to 13 million per day by the end of March 2020.2. Recently, the Taiwan CDC announced regulations requiring all passengers using public transportation to wear face masks all the time and follow the social distancing recommendations.Based on the previous experience of severe acute respiratory syndrome (SARS) in 2003, the Taiwan CDC encouraged residents to wear face masks and perform hand hygiene, especially hospital staff and people in crowded places when the COVID-19 outbreak was initially reported in Wuhan, China in January 20204, and based on the Taiwan CDC data, we believe that a face mask wearing policy also plays an important role in the prevention influenza infection.This successful policy not only controlled the community transmission of COVID-19, but also had an unexpected effect on lowering the incidence of severe complicated influenza after the sixth week this year based on the public records of the Taiwan CDC  and influenza-like illness . Furthermore, even pooled analysis of observational studies revealed the benefit of wearing masks  and respirators  against SARS.Face masks, when fitted properly, effectively disrupt the forward momentum of particles expelled from a cough or sneeze, preventing respiratory disease transmission9 and one meta-analysis reported that the use of face masks to prevent transmission of influenza virus is still controversial10, the mask-wearing policy accompanied with hand hygiene and social distancing appear to have prevented both severe complicated influenza and COVID-19 in Taiwan. In April 2020, the Taiwan Government donated approximately 10 million face masks to countries hit hardest by SARS-COV-2, including the United States and the European Union. We hope that our experience can help other regions to overcome the COVID-19 pandemic as quickly as possible.Although Some studies have been against the use of face masks during the COVID-19 pandemic