Atezolizumab is a programmed death‐ligand 1 (PD‐L1) targeted monoclonal antibody that inhibits PD‐L1 interacting with its receptors PD‐1 and B7‐1, thereby enhancing anticancer immunity. Some real‐world efficacy and safety studies of anti‐PD‐1 antibody have been previously reported. However, there have been no reports investigating the efficacy of atezolizumab monotherapy in clinical practice which have focused on performance status and previous anti‐PD‐1 antibody treatment.We retrospectively reviewed consecutive advanced NSCLC patients who received atezolizumab monotherapy between April 2018 and February 2019 at eight institutions. A total of 152 patients with NSCLC were enrolled in this study.A total of 38 patients (25%) had already been treated with anti‐PD‐1 treatment (nivolumab or pembrolizumab) before atezolizumab. The median OS and TTF was 384 days (12.8 months) , and 42 days (1.4 months) (95% CI: 27–56) in all patients, respectively.p < 0.0001) and TTF  compared with PS 1 or 2–3. Most retreated patients were unable to continue atezolizumab for a longer period, but seven patients (18.4%) were able to continue atezolizumab over four months as an ICI retreatment.ECOG PS 0 had significantly longer OS  continued atezolizumab over four months as an ICI retreatment. Atezolizumab is a programmed death‐ligand 1 (PD‐L1) targeted monoclonal antibody that inhibits PD‐L1 interacting with its receptors PD‐1 and B7‐1, thereby enhancing anticancer immunity.In the OAK trial, atezolizumab showed superiority for overall survival (OS) when comparing docetaxel as a second‐line treatment in patients previously treated with platinum doublet chemotherapy for non‐small cell lung cancer (NSCLC), regardless of PD‐L1 expression.In Japan, atezolizumab was approved for the treatment of patients with advanced NSCLC in January 2018. However, since December 2015, anti‐PD‐1 antibodies (nivolumab and pembrolizumab) were already being used as the standard second‐line treatment in clinical practice for patients with previously treated advanced NSCLC. Some real‐world efficacy and safety studies have subsequently been reported.EGFR mutation status, clinical stage (UICC eighth edition), number of prior regimens, prior ICI therapies, metastatic sites and PD‐L1 expression status before atezolizumab therapy. This study was approved by the institutional review board of each institution. PD‐L1 expression was evaluated by immunohistochemical stain . Atezolizumab 1200 mg/bodyweight was intravenously administered every three weeks until disease progression or unacceptable toxicity.We retrospectively reviewed consecutive advanced NSCLC patients who received atezolizumab monotherapy between April 2018 and February 2019 at eight institutions. A total of 152 patients with NSCLC were enrolled in this study. Clinical data were obtained from medical records according to study protocol. The clinical features included age, sex, Eastern Cooperative Oncology Group performance status (ECOG PS), smoking history, histology, The objectives of this study were to evaluate the efficacy and safety of atezolizumab in patients with advanced NSCLC with endpoints including overall survival (OS), time to treatment failure (TTF) with a data cutoff of August 2020. OS was determined as an interval from the first day of atezolizumab to any cause of death. TTF was defined as an interval from the first day of atezolizumab to the discontinuation of treatment for any reason. Tumor response was evaluated by the investigators according to the Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1. The objective response rate (ORR) was defined as the proportion of patients with the best overall response for complete response (CR) or partial response (PR). Disease control rate (DCR) was defined as the proportion of patients with the best overall response for CR, PR, or stable disease (SD). Adverse events were graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE) version 5.0.t‐test, χ2 tests and Fisher's exact test for continuous and categorical variables, respectively. Statistical analyses were performed using SPSS software, version 23.0 . Statistical significance was indicated by p‐values of less than 0.05.Survival curves were calculated by Kaplan–Meier method and compared by log‐rank test according to ECOG PS and PD‐L1 status. Statistical analyses were performed using Student's EGFR mutation positive patients were enrolled. A total of 38 patients (25%) had already been treated with anti‐PD‐1 treatment (nivolumab or pembrolizumab) before atezolizumab.A total of 152 patients were enrolled in this study for efficacy and safety analyses. The characteristics of all patients are listed in Table At data cutoff (August 2020), 104 patients (68.4%) had died, and 14 patients (9.2%) received continuous atezolizumab treatment. The median follow‐up was 292 days (9.7 months). The median number of cycles of atezolizumab was three. The median OS and TTF was 384 days (12.8 months) (95% CI: 206–424), 42 days (1.4 months) (95% CI: 27–56) in all patients, respectively Figure . In TTF p < 0.0001) and TTF  compared with PS 1 or 2–3  discontinued atezolizumab due to treatment related adverse events (AEs) which included three patients with interstitial pneumonia, one with liver dysfunction, and one with bronchial bleeding. However, there was no treatment‐related death. A total of 55 patients (36.9%) developed immune‐related adverse events (irAEs). Key safety data (irAE) are shown in Table .3% discoTo the best of our knowledge, this is the first and largest multicenter retrospective cohort study to investigate real‐world efficacy and safety of atezolizumab monotherapy in patients with advanced NSCLC. In the present study, ECOG PS was identified as a significantly favorable prognostic factor for OS and TTF. Some previous studies had already demonstrated the same results, but these studies enrolled only ICI naïve patients.The safety of atezolizumab was similar to previous studies. However, liver dysfunction (elevation of AST/ALT) was relatively high in frequency compared to previous prospective pivotal studies. One reason for this might be the enrollment of patients that were heavily treated with other regimens. In Japanese subsets, it is generally well known that interstitial pneumonia is found in higher frequency compared with other races. Recently we reported that honeycomb lung on high resolution CT (HRCT) was a risk factor of ICI‐induced pneumonitis.In Japan, atezolizumab monotherapy was approved as a second‐line treatment for advanced NSCLC in January 2018. However, nivolumab and pembrolizumab had already been established as a standard second‐line treatment at that time. Therefore, some patients were treated with atezolizumab after nivolumab or pembrolizumab in clinical practice. A small case series previously investigated the efficacy of ICI retreatment.EGFR mutant and ICI retreatment patients were enrolled. Therefore, the efficacy might be poor compared with prospective pivotal studies or previous real‐world studies with nivolumab monotherapy. Although the present study revealed that ECOG PS was identified as a significantly favorable prognostic factor for OS and TTF, we were unable to perform multivariate analysis due to the small sample sizes of each ECOG PS subset. Third, some patients were not assessed with regular CT scanning due to clinical practice. Therefore, we could not strictly evaluate progression‐free survival (PFS) and ORR. Finally, a combination regimen of ICI plus platinum doublet chemotherapy has become a standard therapy for first‐line treatment. In patients treated with combination therapy, it might be difficult to apply to our study. However, pembrolizumab monotherapy is still one of the standard regimens for patients with a high expression of PD‐L1 (PD‐L1 ≥ 50%). We expect that our findings could contribute to cases previously treated with pembrolizumab monotherapy as a first‐line treatment.This study has several limitations. First, it was retrospective in design. Second, some In conclusion, in previously treated advanced NSCLC patients, atezolizumab monotherapy demonstrated good efficacy and safety regardless of heavily treated patients in real‐world clinical practice, and ECOG PS 0 was a favorable predictive factor. The efficacy of retreatment with atezolizumab was limited but well tolerated in patients treated with prior anti‐PD‐1 antibody.N. Furuya has received speaker fees as honoraria from AstraZeneca, Chugai, Boehringer Ingelheim Japan, Eli Lilly Japan, Bristol Myers Squibb, Novartis, Taiho, Kyowa Kirin, MSD, and Pfizer.


We applied high-density EEG to examine steady-state visual evoked potentials (SSVEPs) during a perceptual/semantic stimulus repetition design. SSVEPs are evoked oscillatory cortical responses at the same frequency as visual stimuli flickered at this frequency. In repetition designs, stimuli are presented twice with the repetition being task irrelevant. The cortical processing of the second stimulus is commonly characterized by decreased neuronal activity (repetition suppression). The behavioral consequences of stimulus repetition were examined in a companion reaction time pre-study using the same experimental design as the EEG study. During the first presentation of a stimulus, we confronted participants with drawings of familiar object images or object words, respectively. The second stimulus was either a repetition of the same object image  or an image depicting the word presented during the first presentation —all flickered at 15 Hz to elicit SSVEPs. The behavioral study revealed priming effects in both experimental conditions (PR and SR). In the EEG, PR was associated with repetition suppression of SSVEP amplitudes at left occipital and repetition enhancement at left temporal electrodes. In contrast, SR was associated with SSVEP suppression at left occipital and central electrodes originating in bilateral postcentral and occipital gyri, right middle frontal and right temporal gyrus. The conclusion of the presented study is twofold. First, SSVEP amplitudes do not only index perceptual aspects of incoming sensory information but also semantic aspects of cortical object representation. Second, our electrophysiological findings can be interpreted as neuronal underpinnings of perceptual and semantic priming. Bottom-up processing of visual object information goes from simple feature representations Tanaka  to activThe genesis of SSVEPs is not yet explained sufficiently. One theory suggests that SSVEPs can be explained by a linear superposition of transient event-related responses originating in early sensory areas . If SSVEPs were simply an overlap of cortical responses in early visual areas, SR should not result in repetition suppression because both stimuli share no sensory similarities. In contrast, if the two stimuli share certain aspects within their cortical representation, repetition suppression should occur during SR. In other words, if the SSVEP amplitude reflects activity within downstream cortical areas, one has to expect a repetition suppression effect during SR due to the overlapping semantic aspects of the cortical stimulus representations of a word and the representation of its associated picture.The second goal of the present study was to investigate the morphology of the semantic aspects of a cortical object representation as indexed by SSVEP repetition effects. To this end, additionally to SR we used PR by presenting a picture twice. We expected different topographies, in terms of repetition suppression in PR versus SR. Furthermore, we estimated the cortical sources of SR- versus PR-related suppression effects to elaborate on the SSVEP adaptations which are exclusive to SR. In a companion behavioral study, we controlled for the occurrence of behavioral priming effects in an experimental design using flickering stimuli.Nineteen students from Osnabrück University gave their informed consent and participated in the study. Nine of them were males. Their average age was 23.2 years (SD = 2.3 years). They all had normal or corrected to normal vision and no psychological or neurological disorder, specifically they had no migraine or epilepsy and took no medication. The study was approved by the Ethics Committee of Osnabrück University. One data set was excluded due to excessive artifacts in the EEG—therefore, we analyzed the data of 18 participants.We conducted a behavioral companion study to assess priming effects during the presentation of our flickering stimuli. For this behavioral pre-study, we tested 18 participants (different participants as in the EEG experiment were used). One participant was excluded due to migraine after the experiment. All of them were females. Their average age was 21.7 years (SD = 7.5 years).Word , and (6) FillerPicture . Note that only the conditions PP1, PP2, WP2 were used for subsequent analyses. See Table The experimental design was similar as in the publication by Friese et al. . HoweverEach trial (word or picture) consisted of a jittered 500–900-ms baseline period during which a fixation cross was presented, followed by the stimulus for 3000 ms. After the stimulus, a signal cross was presented for 800 ms for the participants to blink if necessary. Participants were asked to avoid eye movements and blinking during the display of the fixation cross and the stimuli. To allow for breaks, the experiment was subdivided into four blocks, each consisting of 120 trials. Prior to the experiment, participants performed a practice block of 80 trials.Every stimulus was located centrally on a black background and covered a visual angle of 7° × 7°, and the fixation cross covered an angle of 3 × 3°, and stimulus onset was synchronized to the vertical retrace of the monitor. We used a monitor with a refresh rate of 60 Hz. The drawings were presented every 4th frame, that is, at 15 Hz, with a duty cycle 1:3, to elicit a 15 Hz SSVEP. This frequency was selected based on a previous study regarding the relation of SSVEPs to object recognition . Eye movements and blinks were measured by vertical and horizontal electro-oculogram. For preprocessing and EEG analysis, we used Matlab 2018a and the EEGLab toolbox version 14  followed by post hoc To receive the temporally changing magnitude of the SSVEP at 15 Hz, the EEG signal was spectrally decomposed by means of Morlet wavelet analysis  were included in the analysis. WP1  was not included due to the mere sensory differences of the stimuli in this condition. All ANOVA-related p values are Greenhouse–Geisser corrected, because in some analyses, the assumption of sphericity was violated.We conducted a 6  × 3  ANOVA with the averaged amplitude as the dependent variable, post hoc one-way ANOVAs, and post hoc t-tests with a significance level of p < 0.05 were performed. Activation threshold corrections accounting for spatial dependencies between voxels were calculated by means of random field theory  which uses the automated anatomical labeling toolbox  = 40.20, p < .001. Post hoc t-tests revealed that participants responded faster to the animate/inanimate judgment in the PP2 condition  than in the WP2 condition , t(16) = 5.4, p < .001, d = 1.3, and in turn faster than in the PP1 condition , t(16) = 4.7, p < .001, d = 1.14, and in turn faster than in the WP1 condition , t(16) = 8.7, p < .001, d = 2.11. The faster responses in the WP2 compared to the PP1 condition shows that semantic priming was successful.In the companion reaction time study, we obtained the expected priming effects. In particular, all reaction times differed significantly depending on the condition, F = 1.76, p = .193.In the EEG study, participants showed an averaged detection rate of 90% and error rates did not differ between conditions, Figure F = 1.80, p = .185. The main effect Regional Mean, F = 13.48, p < .001, F = 13.48, p = .002, F = 0.03, p = .948, left temporal, F = 3.66, p = .054, F = 0.91, p = .404, and right occipital, F = 1.92, p = .167, areas, the condition effect did not reach significance. The SSVEP amplitudes differed sginficantly in the conditions, at the central, F = 7.94, p = .002, F = 5.49, p = .13, In the ANOVA, the main effect Condition did not reach significance, t-tests. Although, strictly speaking, the effect at the left temporal regional mean was not significant, we computed t-tests for this area (1) for explorative reasons, (2) because in source analysis, this regional mean might well play a role in calculating cortical sources for the activation, as well. First, the PP1 versus PP2 comparison reached significance at left occipital, t(17) = 2.47, p = .024, d = 0.58, and at left temporal electrodes, t(17) = − 3.78, p = .001, d = − 0.89. At the left occipital regional mean, the SSVEP response in the PP2 condition  was decreased compared to the PP1 condition . At left temporal electrodes, the SSVEP response in the PP2 condition  was increased compared to the PP1 condition .For each of the significant regional means, we calculated the contrasts of interest, that is, PP1 versus PP2 representing PR effects, and PP1 versus WP2 representing SR effects, for significant differences, via t(17) = 2.97, p = .009, d = 0.70, and additionally at central electrodes, t(17) = 2.67, p = .016, d = 0.63. At both regional means the SSVEP amplitude WP2 condition  was decreased compared to the PP1 condition .Second, the PP1 versus WP2 comparison reached significance at the left occipital regional mean, Figure n = 18, the minimum effect size of d = 0.70 can be detected. Given our effect sizes of d = 0.58 and − 0.89 for PR and of d = 0.70 and 0.63 for SR, one can conclude that the design had enough power to find effects of this magnitude—at least this holds for the effect at the left temporal Regional Mean for the PP1 versus PP2 comparison (d = − 0.89) and for the effect at the left occipital Regional Mean for the PP1 versus WP2 comparison (d = − 0.70). For the one-way repeated-measures ANOVAs, an estimated minimum effect size of f = 0.56, that is, Given a power of 0.80, an α error probability of 0.05, and In the present experiment, we recorded SSVEPs to frequency-tagged word and picture stimuli in a perceptual/semantic stimulus repetition experiment. The first aim of the study was to show that SSVEP amplitudes do not only index the processing of perceptual aspects of incoming sensory information but also semantic aspects of cortical object representations. The second aim was to specify cortical areas that are associated with the processing of semantic features of objects using the SSVEP. In a companion behavioral pre-study, we showed that both perceptual repetition (PR) and semantic repetition (SR) of flickered stimuli are associated with priming effects, indexed by faster reaction times to probe pictures no matter if primed by the same picture  or the corresponding word (semantic priming). Thus, it seems safe to conclude that the present SSVEP results can be interpreted in the light of research dealing with “neuronal priming”, that is, neuronal correlates of implicit mnemonic processing.In particular, we focused on repetition suppression as an index of sharpening processes within the neuronal networks related to perceptual/semantic priming. We found repetition suppression in PR at left occipital electrode sites. These results are in line with findings from an SSVEP study by Martens and Gruber , which eFurthermore, we found repetition suppression in SR. Based on these findings, we conclude that SSVEP amplitude modulations do not only reflect low-level stimulus processing within early sensory areas but also the activation of more widespread cortical networks involved in object recognition. To come to this conclusion, we argue as follows: To observe a repetition suppression effect, specific aspects of a stimulus have to be implicitly remembered during its second presentation. As we observed repetition suppression of the SSVEP during SR, that is, the trials in which a word was used to prime its associated picture, one has to conclude that this neuronal correlate of priming cannot be explained by the repetition of perceptual features. The implicitly remembered stimulus characteristic is its semantic content.In SR, we localized the sources of the repetition effect to bilateral occipital lobes , mainly the right temporal lobe  and smaller parts of the left temporal lobe , bilateral parietal lobes  and the left frontal gyrus. The involvement of frontal regions is in line with a model by Bar et al. which proposes that the prefrontal cortex is also involved in object processing as a “shortcut” and used for top-down processing Bar . FurtherOur results are in line with previous studies, showing that the modulation of the SSVEP signal is not limited to early sensory processes. The fact that SSVEPs reflect the activity of neuronal networks responsible for higher-level processing was, for example, shown in SSVEP studies targeting feature-based attention .In the present study, the stimuli were always presented at 15 Hz. However, previous studies revealed evidence that the driving frequency might have an impact on the pattern of the observed effect. For example, Kaspar et al.  presenteIn summary, our study provides further evidence that SSVEPs reflect the activation of widespread cortical networks associated with the activation of cortical object representations rather than a simple overlap of early sensory processes. This finding provides a solid basis for future studies on object recognition and mnemonic processing, which could exploit the full potential of the SSVEP technique, namely to present the constituting elements of a visual input at different driving frequencies thereby allowing for a separate analysis of the cortical processing of each element of the input (for a seminal example using the multiple-frequency tagging approach see Müller et al.


Venous thromboembolism (VTE) occurs at a high rate after lung cancer surgery and can be attributed to various clinical risk factors. Here, we aimed to determine whether early detection of perioperative D‐dimer and risk‐stratified cutoff values would improve the diagnostic efficacy of VTE.In this case‐control study, D‐dimer results were acquired from 171 non‐small cell lung cancer (NSCLC) patients preoperatively and at the first, third, and fifth day after surgery. VTE was confirmed by Doppler ultrasonography and computer tomography pulmonary angiography (CTPA). Repeated measures ANOVA was used to analyze how D‐dimer changed with time and the effects of risk factors on D‐dimer levels. We then compared sensitivity, specificity and negative predictive value, using both adjusted and unadjusted cutoff values.P < 0.001) due to a trough on the third day, and patients who had undergone thoracotomy (P < 0.001) and those at a more advanced tumor stage (P = 0.037) had higher D‐dimer levels. Area under the curve of D‐dimer was greatest on the third day . Applying stratified cutoff values improved the specificity in the video‐assisted thoracoscopy surgery (VATS) (P = 0.004) and thoracotomy groups (P < 0.001).VTE occurred in 23 patients (13.5%) of the study population. D‐dimer levels increased unsustainably after lung cancer surgery  exclusion. Lung cancer represents as a malignant tumor with high morbidity and mortality worldwide,Considerable efforts have been made to identify the risk factors related with cancer‐associated VTE, and even though the mechanism behind the occurrence of cancer related VTE has not been fully illustrated,This retrospective study reviewed a total of 265 patients newly diagnosed with NSCLC according to pathological results from July 2016 to December 2017, admitted to Beijing Chaoyang Hospital for primary regimen of surgical treatment. A total of 94 patients were excluded for (i) being diagnosed with VTE within three months before surgery; (ii) recent oral administration or subcutaneous injection of anticoagulant agencies; and (iii) incomplete demographic data. A total of 171 patients were enrolled into the study for analysis. None of the included patients received neoadjuvant chemotherapy before surgery. The clinical characteristics and D‐dimer test results required for analysis were obtained from electronic medical records, including age, gender, body mass index, surgical method, surgery procedure, duration of operation, intraoperative blood loss, pathological type, TNM stage and VTE event. A VTE event included deep venous thrombosis (DVT) and pulmonary embolism (PE). Doppler ultrasonography of the lower extremity vein was executed to detect DVT in all patients pre‐ and postoperatively. Computer tomography pulmonary angiography (CTPA) was performed to confirm PE in patients who manifested clinical symptoms or were deemed to be high risk on surgical evaluation. Blood samples were taken at four points of time: the day before surgery, the first, third, and fifth day after surgery. All samples were delivered for D‐dimer assay by immunoturbidimetry  within two hours after collection from veins in the forearm. Surgical options (video‐assisted thoracoscopic surgery [VATS] or open surgery) were determined on the basis of tumor size and location, as well as patients' comorbidities and endurance against relevant surgical procedures. If a difficult situation arose in the process of VATS and traditional thoracotomy was the treatment of choice, then it was manually documented as open surgery in those patients' medical records. All data was collected up to the time of discharge from hospital. The consents were waivered for the retrospective nature of this study. This retrospective study was approved by the Beijing Chao‐yang Hospital Institutional Review Board (ID: 2017‐Ke‐1).P‐value was adjusted by the Bonferroni method. The time effect on D‐dimer levels were described as changes compared with baseline D‐dimer before operation, as the parameter was Ln‐transformed beforehand, so the average increments of D‐dimer concentration (Δ) were described as relative. Receiver‐operating‐characteristic (ROC) curve was used to discriminate VTE patients and non‐VTE patients. The optimum cutoff value was approached by Youden's J statistic. We also calculated sensitivity, specificity and negative predictive value (NPV) according to optimum cutoff values and then used Chi‐square test and Fisher's exact test to compare these parameters between groups. Results having a two‐tailed P‐value <0.05 were considered to be statistically significant. All statistical analyses were conducted using IBM SPSS statistics version 22.0 .The baseline characteristics were expressed as mean ± standard derivation (SD), ratio or median with interquartile range (IQR) accordingly. Multiple imputation was applied when perioperative D‐dimer results were vacant.n = 29, 17%) were squamous cell carcinomas. For pathological staging, there were 127 (74.3%) patients with early stage (0 + I) disease, and 44 (25.7%) with advanced stage (II + III) disease. Until discharge, 23 (13.5%) patients were diagnosed with VTE, including 22 (12.9%) with DVT alone and one (0.6%) with both DVT and PE . The median age at diagnosis was 58 years (range: 35–82 years). The number of male patients was 75 (43.9%) of the whole population. All patients received radical surgical therapy with complete resection. Thoracotomy was performed in a total of 25 (14.6%) patients, one of whom was moved to thoracotomy after the original VATS procedure failed. Most patients had a single pulmonary lobe removed and only two patients underwent pneumonectomy. In terms of histological type, 142 (83.0%) of the tumors were adenocarcinomas and the others (P < 0.001), but not constantly as it met a slight slump on third day, even though it was still higher than that before surgery  . When it came to comparison between groups (Table n = 25) than the VATS group (n = 146) (P < 0.001), and 0 + I stage group (n = 127) had higher levels than those in the II + III stage group (n = 44) (P = 0.037). To be significant, the age distribution between each pair of subgroups was well balanced (see Supplement 1), and there were no significant differences contributed to other clinical factors. Intriguingly, by different grouping, there were the same tendencies of D‐dimer levels as in respective subgroups as in the whole study population , while no significant impact was seen on postoperative D‐dimer levels  Fig . D‐dimertion Fig . At each) P = 0.07. To be P < 0.001), but at the fifth day no significant differences were discovered (P = 0.071). These two subgroups shared a similar growth trend in D‐dimer levels with the whole population. However, in the VTE group (Table P = 0.316), and between the first and fifth day (P = 0.665).In 23 patients (13.5%) diagnosed with VTE after surgery, there were significantly higher levels of D‐dimer in comparison to patients without VTE  on the third day after lung resection, and the cutoff value of D‐dimer was 835 μg/L . The AUC was then calculated separately in different subgroups divided by surgical method and staging. The AUC was higher in the VATS group  than in the thoracotomy group , but with no significant difference . The AUC for 0 + I stage group was 0.784  and 0.705  for II + III stage group, and there was still no significant difference between the two groups . The differences of sensitivity and specificity were not significant between subgroups when using respective cutoff values, as well as when it came to comparison between subgroups using a fixed cutoff value derived from the whole population (Table P = 0.004) and the thoracotomy group (P < 0.001). In terms of NPV, NPV of patients at early disease stage was 97.3% and 96.4% by two kinds of cutoff values (P = 1.000), and in patients with stage II + III, NPV was 91.7% and 91.3%, respectively (P = 1.000). Patients receiving thoracotomy had the lowest NPV  compared with those undergoing VATS .When taking the whole study population into consideration, the area under the curve (AUC) was greatest, which was 0.762  soon after initiation of antitumor treatment for individual patients, so in these cases, the significance of these models is undermined. The Caprini RAM is mainly used for risk stratification and indicating the launch of anticoagulant treatment in a surgical context, and it has been demonstrated to reduce symptomatic VTE by extended prophylaxis within two months after discharge from the thoracic department.Some drawbacks of this study, in particular the small population size and retrospective nature, impeded some conclusive interpretations. The number between subgroups was not equal, which is likely to cause bias. Also, we were unable to develop a risk model because of multicollinearity of the consecutive data and lack of follow‐up. There have been several studies that attempted to identify independent risk factors of VTE in patients who received thoracic surgery for treating lung cancer, but the D‐dimer concentration was enrolled into multivariate analysis with other variables.In conclusion, this study depicted a procoagulant profile by D‐dimer results in NSCLC patients in their early recovery from surgical intervention. The dynamics of D‐dimer in VTE patients represented a differential trend from others, but there were no significant differences of trend between patients of different tumor stage and different surgical options, except for quantitative imparity. However, using stratified rather than collective cutoff values is of benefit for the exclusion and diagnosis of VTE events in lung cancer patients in the early days after curative resection.All authors declare that there are no conflicts of interest.AppendixS1: Supporting InformationClick here for additional data file.


The physicochemical properties of flavonoids suggest they could function as mitochondrial uncouplers. Moreover, mitochondrial uncoupling alleviated neurodegeneration in Caenorhabditis elegans during aging in previous research. Therefore, we investigated whether various flavonoids  could reduce neuronal defects by mitochondrial uncoupling in C. elegans. Both neuronal defects and mitochondrial membrane potential were reduced in aged worms in nearly all of the flavonoid treatments suggesting that flavonoids may reduce neurodegeneration in C. elegans. However, there was no significant reduction of neuronal defects in mitophagy‐deficient pink‐1/pdr‐1 double mutants under flavonoid treatments. These results suggest that flavonoids could function as mitochondrial uncouplers to mitigate neurodegeneration in aged C. elegans, possibly via a PINK1/Parkin mitophagy process.Dietary supplementation of flavonoids has been shown to reduce the severity of neurodegenerative disorders such as dementia, Parkinson's disease, and Alzheimer's disease by their antioxidant effects. However, their low bioavailability C. elegans in association with reduced mitochondrial membrane potential, indicated by TMRE staining. The results were reproduced in uncoupling mutant, ucp‐4, and chemical uncoupler DNP treated worms, but not in pink‐1;pdr‐1 mutant worms. Therefore, flavonoids could function as a mitochondrial uncoupler to mitigate neurodegeneration in aged C. elegans, possibly via a PINK1/Parkin mitophagy process. Flavonoid treatment reduced neurodegeneration in aged  PTEN‐induced putative kinase 1 (PINK1) protein in mitochondria senses mitochondrial membrane depolarization, and Parkin amplifies this signal to initiate mitophagy , pdr‐1 , and zdIs5 , were provided by the Caenorhabditis Genetics Center (CGC) at the University of Minnesota. pink‐1 deletion mutant (tm1779) was provided by the Mitani laboratory through the National Bio‐Resource Project (Japan). For observation of mechanosensory neurons, all strains were crossed with zdIs5. The ucp‐4 mutant was crossed with zdIs5 to obtain zdIs5;ucp‐4. The pink‐1 mutant was crossed with zdIs5 to obtain zdIs5;pink‐1, which was further crossed with pdr‐1 to obtain pink‐1;pdr‐1 double mutants.The strains, Bristol N2 wild‐type, C. elegans and prevent internal egg hatching, fluorodeoxyuridine (FUdR) was applied to the top of the NGM‐OP50 plates at a final concentration of 50 µM. Worms at L4 stage were transferred to the FUdR‐treated plates.The culture conditions were in accordance with the standard protocols Brenner, . For rel2.2p‐values < .05 were considered statistically significant.Neuronal defects were determined by observation of anterior lateral microtubule (ALM) and posterior lateral microtubule (PLM) morphological abnormalities, specifically an outgrowth in ALM cells, and/or wavy, neuronal sprouts and branching in PLM cells  of DNP  in dimethyl sulfoxide  was added to NGM medium at the desired final concentration before pouring into plates. The flavonoids fisetin, quercetin, apigenin, chrysin, catechin, and naringenin, were obtained from Cayman Chemical Company . Flavonoid‐NGM plates were prepared using the same procedure mentioned for DNP. Worms at L4 stage were transferred to flavonoid‐containing NGM plates.2.4m) owing to its cell permeability and cationic fluorescence . One‐way ANOVA analysis was performed, followed by an unpaired t test, for DMSO treatment and flavonoid treatments in worms.Tetramethylrhodamine ethyl ester  is used as an indicator of mitochondrial membrane potential (Ψ33.1m) by functioning as mitochondrial uncouplers. Flavonoids are classified into four subclasses depending on the degree of unsaturation and oxidation of the C ring: flavonols, flavones, flavanols, and flavanones  to investigate the effects of flavonoids on aging‐associated neurodegeneration. According to Koch et al., flavonoid concentrations in the range of 0.1–200 µM resulted in increase the mean life span in C. elegans . Quercetin‐treated worms showed 32% less ALM and 31% less PLM neuronal defects at day 5, and 30% less ALM and 21% less PLM neuronal defects at day 10 as compared to the DMSO‐treated worms . The ALM neuronal defect was 26% less in apigenin‐treated worms than in the DMSO control worms at days 5 and 10 . The intensity of staining was less in worms treated with 100 µM DNP than that of the DMSO‐treated controls, which is consistent with our previous study . These results are consistent with the results of our previous study . The low TMRE intensity in aged worms could be due to reduced TMRE uptake and accumulation, since TMRE accumulation in mitochondria depends on mitochondrial activity  was obtained for quercetin‐treated N2 and s Figure . Quercets Figure . Moreoves Figure . Taken tRecent research showed that catechin also inhibits mitochondrial complex I, resulting in reduced mitochondrial membrane potential and less hydrogen peroxide production in isolated mitochondria  single mutants. Both pink‐1 and pdr‐1 mutants showed no significant difference in neuronal defects with flavonoid treatments and DMSO treatments (data not shown). Therefore, we used the pink‐1;pdr‐1 double mutant in the zdIs5 background to investigate whether mitophagy was involved in flavonoid‐mediated alleviation of neurodegeneration in aged C. elegans. First, we confirmed that there was no significant difference in the number of ALM and PLM defects between pink‐1;pdr‐1 double mutants and zdIs5 control worms at days 1–15 (data not shown). No significant differences in ALM and PLM neuronal defects were observed between flavonoid‐treated and DMSO‐treated worms at day 5  flavonoids act as mild mitochondrial uncouplers to dissipate mitochondria membrane potential, and (2) by doing so, flavonoids reduce neurodegeneration in aged The authors declare no conflict of interest.This study does not involve any human or animal testing.Table S1‐S4Click here for additional data file.


The aragonite shelled, planktonic gastropod family Atlantidae (shelled heteropods) is likely to be one of the first groups to be impacted by imminent ocean changes, including ocean warming and ocean acidification. With a fossil record spanning at least 100 Ma, atlantids have experienced and survived global-scale ocean changes and extinction events in the past. However, the diversification patterns and tempo of evolution in this family are largely unknown.c oxidase subunit 1 mitochondrial DNA, 28S and 18S ribosomal rRNA) we show that the three extant genera of the family Atlantidae, Atlanta, Protatlanta and Oxygyrus, form monophyletic groups. The genus Atlanta is split into two groups, one exhibiting smaller, well ornamented shells, and the other having larger, less ornamented shells. The fossil record, in combination with a fossil-calibrated phylogeny, suggests that large scale atlantid extinction was accompanied by considerable and rapid diversification over the last 25 Ma, potentially driven by vicariance events.Based on a concatenated maximum likelihood phylogeny of three genes (cytochrome Now confronted with a rapidly changing modern ocean, the ability of atlantids to survive past global change crises gives some optimism that they may be able to persist through the Anthropocene. Atlanta, Protatlanta and Oxygyrus) have thin-walled laterally compressed aragonite shells that are broadened with a keel. A modified foot serves as a primary swimming fin, and the broad shell is used as a secondary swimming fin [The Atlantidae is a family of small < 14 mm) marine predatory gastropods with a holoplanktonic mode of life , and A. plana in the last 3.5–1 ka. Protatlanta sculpta Issel, 1911 is currently only known from the Atlantic Ocean, but was present in Late Pleistocene sediments of the Indian Ocean 24–16 ka ago  and in Pliocene rocks of Pangasinan, Philippines [The thin-walled aragonite shells of the atlantids, and their habitat in the upper ocean imply that they are likely to be sensitive to ocean acidification and ocean warming, in a similar way to the shelled pteropods . The onlic Ocean , howeveric Ocean , and A. rn Spain , 14. Thelippines . Most ofCoelodiscus, Freboldia and Tatediscus [Coelodiscus minutus  [Coelodiscus minutus has a shell morphology remarkably similar to larval atlantid shells of the genus Atlanta  , 21 frometeropod . The plaed later .Fig. 2CBellerophina minuta  [Oxygyrus  is now also placed [Oxygyrus and B. minuta remains enigmatic [B. minuta to belong within the family Atlantidae, having a shell morphology very similar to the extant genus Oxygyrus [Bellerophina within the Family Atlantidae. Freboldia fluitans Nützel & Schneider, 2016 (163.5–157.3 Ma) is thought to be holoplanktonic and its shell morphology is involute and quite rounded in shape, however, unlike B. minuta, the shell surface has little or no ornamentation, and the coiling direction of F. fluitans cannot be determined with certainty [The oldest potential member of the family Atlantidae does not appear in the fossil record until ~ 57 Ma later, in the Early Cretaceous. y, 1812) , 23, fouestombes  considernigmatic , and herOxygyrus . Indeed,Atlantidea [Through the Late Cretaceous, Paleocene and Eocene there are no known atlantid fossils, creating a gap in the record of ~ 73 Ma. The recent fossil record of the Atlantidae, extending to the Piacenzian (3.6–0 Ma) is relatively well known . Howeverlantidea .Atlanta, with an entirely aragonite shell, is the earliest diverged, and that the genus Oxygyrus, with a shell composed of both aragonite and conchiolin (probably to improve buoyancy), is the latest to diverge [B. minuta is morphologically most similar to Oxygyrus  sequences from specimens of all known atlantid morphospecies. However, deeper genus level relationships were not resolved in this study, and it provided few clues about the longer-term evolutionary history of the family. In the present study, an extended CO1 dataset is used in combination with two nuclear genes, 28S and 18S, to produce a more complete molecular phylogeny with which to compare previous morphology based hypotheses of atlantid evolution. Through a fossil-calibration of this phylogeny, the likely timing of diversification reveals the persistence of this successful group of holoplankton through past ocean changes.Only a single large-scale study has previously explored the molecular phylogeny of the atlantid heteropods . Wall-PaA. peronii C, and one within O. inflatus A). This brings the total number of putative species to 36, of which 24 are described as morphospecies and 12 are undescribed putative species. However, the deeper relationships between atlantid genera were not supported within the CO1 phylogenetic tree  alignment of CO1, 28S and 18S recovered species and genera, with node supports of > 80% (100% for most) at most levels of the atlantid tree . The A. peronii and A. gibbosa groups are not monophyletic due to the position of A. frontieri Richter, 1993, which falls within the A. gibbosa group . The larger and non-ornamented shelled species are grouped together, but have low node support (58% support). These two broad groups are also supported by radula type . The calibration fossil for the oldest member of the superfamily Pterotracheoidea (heteropods) roots the molecular clock analyses within the Early Jurassic  in the Oligocene. However, the partial replacement of aragonite with conchiolin in the shells of this lineage may have been a response to ocean acidification. This change is particularly evident in Oxygyrus where conchiolin covers the aragonite shell entirely, likely protecting it from seawater chemistry in a similar way to the periostracum in pteropods [Due to the limited atlantid fossil record, it is not possible to determine whether the atlantids diversified during the Late Cretaceous, Paleocene or Eocene, only that the ancestor of teropods . All majteropods , 42, expteropods . It is tteropods , possiblteropods .Atlanta during the last ~ 25 Ma  of the Mid-Miocene at ~ 18–12 Ma  and the Atlanta peronii A, B, C1 and C2, Atlanta rosea A and B Gray 1850, Atlanta meteori A and B Richter, 1972, Atlanta oligogyra B and C Tesch, 1906  show clear geographical separation, with one putative species being present in the Atlantic Ocean only, and the other being present in the Pacific and Indian Oceans [Atlanta cordiformis Gabb, 1873 at ~ 5.33 Ma [Atlanta at this time was probably also accompanied with population bottlenecks and  extinctions caused by ocean changes related to the IoP. More recently, localised extinctions of Atlanta turriculata and Atlanta plana in the Atlantic Ocean, and Protatlanta sculpta in the Indian Ocean have been detected in the Quaternary fossil record [Molecular dating analyses indicate that most modern species arose following the TTE in the Tortonian at ~ 12–10 Ma Fig. . Additio906 Fig. . This man Oceans . A seconl record . These eCalcifying plankton are widely accepted to be amongst the most sensitive and first affected by current and predicted future warming and acidification of the world oceans . The res2 induced global warming) become available to organisms at unusually high rates, there are increased opportunities for evolution and diversification. Planktonic organisms have high evolutionary potential and are considered to be well poised for evolutionary responses to global change [The present study shows that although global-scale environmental changes over the last 25 Ma have likely caused large scale extinction of atlantids, they have also resulted in periods of exceptional diversification. Past ocean changes, largely resulting from vicarance events, provided opportunities for speciation and have driven evolution within this family. Vermeij  hypothesl change . TherefoN = 28) and 2017 . A total of 24 specimens were collected from nine stations in the Indian Ocean during oceanographic cruises SN105 (N = 18) and VANC10MV (N = 6). In the Pacific Ocean, 42 specimens were collected from 23 stations during cruises ACE-ASIA (N = 1), DRFT (N = 2), KH1110 (N = 16), KM1109 (N = 2), KOK1703 (N = 3), S226 (N = 4), SO255 (N = 13) and WCOA16 (N = 1). In addition, for the concatenated gene phylogeny, two specimens of the partially shelled heteropod genus Carinaria, collected during the oceanographic cruise SN105 (N = 2), were used as outgroup taxa.A total of 588 specimens from all 34 known putative atlantid species , includiSonne to the north east of New Zealand between March and April 2017. Specimens were collected using a ring net with an aperture of 1 m diameter, a mesh size of 350 μm and a maximum sampling time of 30 min. At stations SO255_041 and SO255_057, vertical net hauls were carried out from 200 m water depth to the surface. At all other stations, oblique tows were made in the upper 100 m. Cruise KOK1703 took place offshore of Hawaii, around station ALOHA, on board the RV Ka’imikai-O-Kanaloa in March 2017. Sampling was carried out using either a 0.71 m diameter CalBOBL bongo net with a mesh size of 200 μm, or a ring net with a 2 m diameter and mesh size 505 μm. For all stations, oblique tows were conducted in the upper 200 m for a maximum of 44 min. During cruise AMT27, specimens were collected using a 0.71 m diameter bongo net and a 1 m diameter ring net, both with a mesh size of 200 μm. Oblique bongo net tows sampled a range of maximum depths from 233 to 388 m and ring net tows sampled a range of maximum depths from 56 to 99 m.Collection methods included the use of a variety of plankton nets . These have been previously described for most of the oceanographic cruises listed here , 50–54. Prior to DNA extraction, all specimens were imaged using stacking microscopy on a Zeiss Discovery V20 or V12 microscope  was amplified using primers jgLCO1490 (5′–TITCIACIAAYCAYAARGAYATTGG–3′) and jgHCO2198 (5′–TAIACYTCIGGRTGICCRAARAAYCA–3′) [Three commonly used gene fragments were amplified. A ~ 570 bp fragment of the mitochondrial cytochrome AYCA–3′) . A ~ 100AYCA–3′)  and D3-RAYCA–3′)  and 1800AYCA–3′) .All PCR reactions contained 17.30 μl ultra pure water (mQ), 2.50 μl 10x Qiagen PCR buffer, 0.50 μl 25 mM MgCl2, 1.00 μl 100 mM BSA, 1.00 μl 10 mM of each primer, 0.50 μl 2.5 mM dNTPs and 0.25 μl 5 U Qiagen Taq, with 1.00 μl of template DNA, which was diluted up to 100 times for some samples. The same PCR steps were performed for all three genes using an initial denaturation step of 180 s at 96 °C, followed by 40 cycles of 15 s at 96 °C, 30 s at 50 °C and 40 s at 72 °C, and finishing with a final extension of 300 s at 72 °C. Sequencing of forward and reverse strands was carried out by either Macrogen Europe (Amsterdam), or Baseclear (Leiden).Carinaria from the closely related heteropod family Carinariidae was used as an outgroup with 14 CO1, three 28S and four 18S sequences included from 15 specimens collected in the Indian Ocean. Here, we consider a ‘putative species’ to be a well-supported monophyletic group (bootstrap support > 80%) in our CO1 phylogenetic tree, with interspecific genetic distances of > 6.95% (Jukes-Cantor). We verified our selection of putative species using Automatic Barcode Gap Discovery (ABGD) performed on the entire CO1 dataset using Jukes-Cantor genetic distances with default settings [Atlanta inclinata and Atlanta tokiokai van der Spoel & Troost, 1972 were grouped together due to high intraspecific genetic diversity. However, here we consider them to be two separate species because of clear morphological differences. All other previously identified putative species [A. peronii C, and one within O. inflatus A, bringing the total number of putative species to 36.For individual gene phylogenies, a total of 237 new 28S sequences, 230 new 18S sequences, and 99 new and 477 previously published , 59, 60 settings . Similarsettings , Atlanta species  were recAtlanta plana and Atlanta selvagensis, it was necessary to combine genes from different specimens at the same station , in order to obtain complete taxon sampling for all genes. Where records have been combined, specimens were confirmed to be conspecific using CO1, which was available for all specimens. All three markers were obtained for each putative species in each region, except for Atlanta gaudichaudi, which was only successful for CO1 and 18S, and Atlanta californiensis for which only CO1 was sequenced. A total of 102 CO1, 100 28S and 101 18S sequences are included in the concatenated gene phylogeny.For the concatenated gene (3-gene) phylogeny, at least one specimen from each atlantid putative species was included from each ocean basin in which that putative species resides. As far as possible, all three genes were combined from a single specimen. In two cases, for New sequences were verified and edited using Geneious R8 and all sequences were aligned using MEGA 7 . All gapSingle gene and 3-gene phylogenetic relationships were resolved using maximum likelihood analyses in RaxmlGUI 1.5b2 . Using jCarinaria concatenated gene sequences were included in the fossil-calibrated phylogeny, including a single representative of each putative species identified in previous CO1 phylogenies [Coelodiscus minutus [Bellerophina minuta [Atlanta . Running the Bayesian analysis without the Atlantidea rotundata calibration changed the topology of the tree, grouping Atlanta with Oxygyrus. This placement is not in agreement with the topology of the concatenated ML phylogeny, which shows that relationships between the three genera are not well resolved with the current molecular dataset. Finally, the morphospecies Atlanta peronii, which is made up of four putative species, was used to calibrate more recent speciation [Freboldia (163.5–157.3 Ma) was not included in the calibrations because, unlike B. minuta, this species does not show morphological similarity to the Atlantidae and we are unconfident in its current placement [8 generations each and checked for convergence with a burnin of 10% in Tracer 1.6.0 [4 generations. Maximum clade credibility trees were selected using TreeAnnotator 2.5.0 [A subset of 34 atlantid and one logenies   and theanta sp. ) and Proiraensis ) were uskoroidea , 66. Basotundata  was usedeciation . The foslacement . Two inder 1.6.0  before ber 1.6.0 . Trees aor 2.5.0 . CalibraAdditional file 1: Supplementary Table 1. Specimens included in this study.Additional file 2: Supplementary Figure 1. Maximum likelihood phylogeny of the family Atlantidae based on cytochrome c oxidase subunit 1 mitochondrial DNA (CO1). Black squares represent bootstrap support > 80%. Species groups based on morphology are highlighted with coloured boxes  have been collapsed to simplify the phylogeny. Black squares represent bootstrap support > 80%. Species groups based on morphology are highlighted with coloured boxes  have been collapsed to simplify the phylogeny. Black squares represent bootstrap support > 80%. Species groups based on morphology are highlighted with coloured boxes .


Aquila audax fleayi), an endangered Australian raptor and facultative scavenger, showed evidence of lead exposure. We detected lead in 100% of femur and liver tissues of 109 eagle carcasses opportunistically collected throughout Tasmania between 1996 and 2018. Concentrations were elevated in 10% of 106 liver (>6 mg/kg dry wt) and 4% of 108 femur (>10 mg/kg dry wt) samples. We also detected lead in 96% of blood samples taken from 24 live nestlings, with 8% at elevated concentrations (>10 μg/dL). Of the liver samples with elevated lead, 73% had lead207/206 isotope ratios within the published range of lead‐based bullets available in Tasmania. These first comprehensive data on lead exposure of an Australian raptor are comparable to those for raptor studies elsewhere that identify lead‐based ammunition exposure as a conservation threat. Our findings highlight the importance of further research and efforts to address lead contamination throughout the Tasmanian ecosystem and in other Australian regions. Environ Toxicol Chem 2021;40:219–230. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.Lead poisoning, mainly through incidental ingestion of lead ammunition in carcasses, is a threat to scavenging and predatory bird species worldwide. In Australia, shooting for animal control is widespread, and a range of native scavenging species are susceptible to lead exposure. However, the prevalence of lead exposure in Australia's scavenging and predatory birds is largely unknown. We evaluated the degree to which the Tasmanian wedge‐tailed eagle ( Lead is a toxic metal that can negatively affect a range of physiological systems, thereby threatening susceptible animal populations and ecosystems  and 400 000 brushtail possums (Trichosurus vulpecula) are shot annually across the Australian island state of Tasmania  shares the same characteristics of avian scavengers on other continents that are threatened by lead exposure. Recent preliminary work has demonstrated that wedge‐tailed eagles on mainland Australia are exposed to lead  is a subspecies endemic to the island of Tasmania. The population is listed as endangered at both state and federal levels . Second, we drew blood from live‐caught nestlings. The present study was conducted in accordance with the University of Tasmania Animal Ethics Committee Permit A0015930 and Department of Primary Industries, Parks, Water and Environment scientific permits TFA17127 and TFA17328.Carcasses had been collected opportunistically by government departments, industry representatives, and volunteers, between 1996 and 2018, and stored frozen by the Department of Primary Industries, Parks, Water and the Environment  and the Tasmanian Museum and Art Gallery . Between May 2017 and March 2018, we defrosted carcasses and harvested tissue. At necropsy, we collected a liver lobe, sectioned a femur diaphysis (≈3 cm length from the middle of the femur), and estimated the bird's age at death.Nestlings were sampled prior to fledging (estimated age 65–75 d) during 2017 and 2018. We selected target nests identified during fixed‐wing aerial nest surveys  from each sample using a stainless‐steel scalpel blade. We then sectioned target tissue from the middle of each liver lobe (≈1.3 g) and femur diaphysis (≈460 mg). We dehydrated femur and liver samples at 60 °C until a constant weight was reached. Dried samples were weighed on a digital balance . A 1‐mL volume of each blood sample was also dehydrated and weighed using the same method. We stored samples in metal‐free plastic containers until digestion.Samples were analyzed for lead concentration and isotope ratios at Edith Cowan University Analytical Facility . Liver and bone samples were homogenized, and 0.4 g was aliquoted into inductively coupled plasma (ICP)‐grade Teflon vessels containing 5 mL of trace analysis–grade nitric acid 68% , 0.5 mL of trace metal–grade hydrochloric acid 34 to 37% (Fisher Scientific), and 3 mL of hydrogen peroxide 30% . Samples were digested for 15 min in a Multiwave GO microwave digestion system (Anton Paar) set to 150 °C. After digestion, samples were diluted to 50 mL with Milli‐Q® reverse osmosis deionized water and transferred to polypropylene tubes for analysis. Blood samples were prepared by a similar method using digestion in 2 mL of nitric acid, 0.2 mL of hydrochloric acid, and 1 mL of hydrogen peroxide. Blood sample solutions were then sonicated at 60 °C for 1 h and diluted to 10 mL using Milli‐Q® reverse osmosis deionized water.Lead concentrations were determined via ICP–mass spectrometry (MS) using an iCAP Q ICP‐MS (Thermo‐Fisher Scientific) coupled to an ASX‐520 AutoSampler (Agilent Technologies). Data acquisition, element quantitation, and isotope percentage analyses were carried out using Qtegra (Thermo‐Fisher Scientific). The instrument was calibrated using concentration ranges of iCAPQ element standards (Thermo‐Fisher Scientific) and ICP–MS‐68A solutions (High Purity Standards) to provide standard curves before analysis. Certified reference materials (CRMs) were used as positive controls for each tissue type. These were Bone Ash Standard Reference Material 1400  and Bovine Liver Certified Reference Material BCR–185R . Two digestions were carried out on each CRM with 2 ICP–MS readings of each digestion. Accuracy of CRM ICP–MS readings averaged 96.7% for femur and 104.1% for liver. Every 10th sample was re‐analyzed for a duplicate read (average relative standard deviation [RSD] 1.8%), and duplicate blind sample digestions were carried out for 20 randomly selected samples (average RSD 5.5%). Lead concentrations were measured as mg/kg dry weight. Limits of quantification and limits of detection (LOD) for the analysis were 0.005 and 0.0015 mg/kg, respectively.207/206 ratio. This ratio is used most commonly in research investigating sources of lead exposure in birds  measured at m/z 204, 206, 207, and 208. Isotope readings were adjusted according to readings from a calibration lead solution at m/z 204 (1.40%), m/z 206 (24.10%), m/z 207 (22.10%), and m/z 208 (52.40%). Analyses of isotope data focused on the leadTo interpret liver lead concentrations in terms of their potential physiological effects, we used previously identified thresholds of <6 mg/kg dry weight as evidence of low levels of exposure with limited health implications  to wet weight (μg/dL) by multiplying the dry weight concentrations by the dry to wet weight ratios of each sample. We used an exposure threshold of blood lead <10 μg/dL as indicative of low exposure  for liver and femur lead concentrations of all samples and also for each age class. In the case of the blood lead concentrations, for which some samples had values below the LOD of the ICP–MS , we used a Kaplan–Meier cumulative probability distribution  between tissue lead levels and the distance between samples.Eagle carcasses were not collected, nor were nestlings identified and sampled, in a spatially random manner. Thus we used Mantel tests  and those with low concentrations (<6 mg/kg) using a modified signed‐likelihood ratio test  by assessing differences in leadn = 71; MRM: R2 = 0.003, F = 8.382, p = 0.393; Mantel: p = 0.721, r = –0.029), femur , or nestling blood samples .We collected tissue from 109 eagle carcasses, of which 27 were adults and 82 pre‐adults. We analyzed tissue lead concentrations in 108 femur samples (26 adults and 82 pre‐adults) and 106 liver samples (26 adults and 80 pre‐adults). The location of carcass recovery was recorded for 72 eagles, and the date of recovery was recorded for 61. We collected blood samples from 24 eagle nestlings. There was no evidence of spatial organization in lead concentrations of liver . However, the 6 highest liver lead concentrations (range: 43.9–181.6 mg/kg) were markedly higher than the highest femur lead concentration (25.6 mg/kg). Lead concentrations in femur and liver samples from the same individual were weakly correlated with each other .Lead concentrations in femur samples were significantly higher than in liver samples  in 23 of 24 nestling blood samples  than adults . Lead concentrations of femurs from adult birds were significantly higher than those from pre‐adult birds  than for pre‐adults . However, the only bird with femur lead concentrations indicative of severe exposure (>20 mg/kg) was a pre‐adult.Lead concentrations in livers were similar in both age groups . That said, elevated and severe liver lead concentrations were only recorded in winter, spring, and summer, and never in autumn.We detected no interseasonal difference in liver lead concentrations . Because the sample size was smaller in this test and because residuals indicated poor matching to distributional assumptions, we interpreted only the Wilcoxon tests.207/206 isotope ratios within liver samples were 0.8835 ± 0.0539 . The lead207/206 ratios of eagles with elevated hepatic lead concentrations  were significantly different from those with low lead concentrations , despite the disparity in sample size, which reduces power to detect a difference. The level of variation in lead207/206 isotope ratios was similar between these 2 groups . Lead207/206 isotope ratio was negatively correlated with liver lead concentration  and correlative conclusions . For nestling blood samples with lead (n = 23), lead207/206 ratios were not correlated with blood lead levels . The low number of nestling blood samples with elevated lead levels (n = 2) precluded statistical comparisons of lead207/206 ratios between different blood lead thresholds.Mean lead207/206 isotope ratios overlapped the published range of lead207/206 ratios for a subset of brands of lead‐based bullets available in Tasmania (0.8070–0.9561). The majority (72.7%) of liver samples with elevated lead concentrations  had lead207/206 ratios that were within this reported isotopic range of ammunition. The majority (91.3%) of nestling blood samples with detected lead (n = 23) also had lead207/206 ratios that were within the published isotopic range of ammunition brands available in Tasmania.Liver leadOur study provides the first comprehensive data on lead exposure of an Australian raptor species and highlights the value of analyzing different tissues when assessing the extent of lead contamination. Whereas a low proportion of femur samples had elevated lead concentrations, liver concentrations showed more substantial evidence of lead exposure. The higher lead concentrations detected in femurs of older birds compared with femurs from pre‐adults suggests that birds are chronically exposed. The absence of seasonal trends in hepatic lead suggests that risk of exposure may not change throughout the year. Lastly, the isotope ratios are consistent with an ammunition source of contamination, but additional information on the isotope ratios of other lead sources would be useful to clarify routes of exposure in Tasmania.Aquila chrysaetos; 0.4–54.21 mg/kg, n = 46; Ganz et al. Coragyps atratus; 4.5–540 mg/kg, n = 98; Behmke et al. Aquila adalberti; <LOD–41.68 mg/kg, n = 84; Rodriguez‐Ramos Fernandez et al. n = 11). The maximum bone lead concentrations we detected are similar to concentrations observed in studies of golden and bald eagles  lies within the range of estimates for other avian scavengers threatened by lead poisoning  is higher than those found in congeners  represents a likely lethal dose  of nestlings we tested had been exposed to lead. Although this rate is higher than that reported for other raptors Harmata , that moAlthough our carcass sampling was opportunistic, the absence of seasonal differences in lead exposure of Tasmanian wedge‐tailed eagles likely reflects the near absence of seasonality in local ammunition use. An increase in lead exposure during designated hunting seasons has been reported in scavenging bird populations in Europe (Ecke et al. The range in isotope ratios that we detected was much greater than those documented for other avian scavengers (Finkelstein et al. 207/206, there are concerns over the precision of lead207/206 estimates obtained by this method (Gulson et al. Our ability to link the lead exposure detected in the Tasmanian wedge‐tailed eagle to lead‐based ammunition was limited in 3 ways. First, although recent work has frequently used Q ICP–MS to measure lead207/206 isotope ratios within the range of ammunition was comparable to other studies implicating ammunition as a source of lead poisoning (e.g., 79%; Finkelstein et al. 207/206 ratios associated with ammunition (Church et al. Despite the ambiguity of our isotopic data, they do provide some evidence that at least some of the lead exposure we recorded arose from consumption of ammunition. This is because, first, the proportion of individuals with leadThe physiological effects of different levels of lead exposure can be difficult to interpret because of the paucity of experimental evidence quantifying the effects of tissue lead concentrations. Lead concentrations we have categorized as “low” do not equate to no physiological effect (Ganz et al. Our results indicate that lead exposure is likely a significant factor to consider in conservation management for the Tasmanian wedge‐tailed eagle. Although some Australian eagles are known to die directly from lead poisoning Mooney , lead maSarcophilus harrisii; Hivert et al., Our study highlights the importance of further research and efforts to address lead contamination throughout the ecosystem and in other Australian regions. In Tasmania, high levels of lead exposure are unlikely to be confined to wedge‐tailed eagles. In fact, numerous species are known to scavenge shot carcasses, including the endangered Tasmanian devil (https://doi.org/10.1002/etc.4914.The Supplemental Data are available on the Wiley Online Library at Any use of trade, product, or firm names is for descriptive purposes only and does not imply endorsement by the US Government.J. Pay, T. Katzner, J. Wiersma, C. Hawkins, A. Koch, and E. Cameron conceived the study. W. Brown and N. Mooney collected carcasses. J. Pay collected tissue samples from carcasses. J. Wiersma collected blood samples. J. Pay prepared all samples for laboratory analysis. J. Pay analyzed the data with assistance from T. Katzner. J. Pay led writing of the manuscript, and all co‐authors contributed to revisions of the manuscript.This article includes online‐only Supplemental Data.Supporting information.Click here for additional data file.


Anterior decompression with fusion (ADF) has often been performed for degenerative cervical myelopathy (DCM) in patients with poor cervical spine alignment and/or anterior cord compression. We aimed to identify clinical and radiological predictors associated with neurological recovery after ADF.This post-hoc analysis from a prospective multicenter study included patients who were scheduled for ADF for DCM. The patients who received other surgeries  were excluded. The associations between baseline clinical and radiographic variables  and the recovery rates as the outcome variables were investigated in the univariate regression analysis. Then, the independent predictors for increased recovery rates were evaluated using a stepwise multiple regression analysis.p = 0.049), SVA  and PCS  score were significantly associated with recovery rate. Then, a stepwise multiple regression analysis identified the independent predictors of recovery rate after ADF as TK (p = 0.01), PCS (p = 0.03), and SVA (p = 0.03). According to this prediction model, the following equation was obtained: recovery rate = − 8.26 + 1.17 × (TK) − 0.45 × (SVA) + 0.85 × (PCS) .In total, 37 patients completed the 1 year follow-up. The recovery rate was significantly correlated with SVA (p = 0.001) and TPA (p = 0.03). Univariate regression analyses showed that age (Regression coefficient = − 0.92, Patients with lower TK, lower PCS score, and higher SVA were more likely to have poor neurological recovery after ADF. Therefore, patients with DCM and these predictors who undergo ADF should be warned about poor recovery and be required to provide adequate informed consent. With age, degeneration of the cervical spine progresses . AccordiTo date, research regarding preoperative predictors for the success of ADF has been limited. A prospective randomized study showed that preoperative predictive factors of good outcome 10–13 years after ADF included initial high neck-related pain intensity, nonsmoking status at the time of surgery, and male sex . A retroThe aim of this study was to identify clinical and radiological predictors, including whole spinal radiographic parameters, associated with neurological recovery after ADF.This study was a secondary analysis of a previous prospective multicenter study that investigated the characteristics of patients with DCM and their surgical outcomes . BrieflyThe current study included patients with DCM who had undergone ADF. Accordingly, patients who received other surgeries  were excluded Fig. . This stAs radiological parameters, we measured cervical lordosis (CL), C2-7 range of motion (ROM), C7 slope, C2-7 sagittal vertical axis (SVA), thoracic kyphosis (TK), lumbar lordosis (LL), sacral slope (SS), SVA, pelvic tilt (PT), and t1 pelvic angle (TPA).CL was defined by the Cobb angle between C2 and C7 on a lateral radiograph in the neutral position. C2-7 ROM was measured on flexion–extension lateral radiographs. The C7 slope was calculated by measuring the angle formed by the horizontal line to the C7 vertebra and the line parallel to the superior endplate of the C7 vertebra . TK was Outcomes were assessed before surgery and 1 year after surgery using the Japanese Orthopedic Association score for the assessment of cervical myelopathy  , the Eurp-value < 0.25 in univariate regression analyses were carried forward to the second step of the analysis [We performed a paired two-tailed t-test for normally distributed data or a Wilcoxon signed-rank test for skewed distributed data to identify differences in scores before surgery and 1 year post-surgery after assessing normality with the Shapiro–Wilk test. Spearman correlation coefficients were used to evaluate the relationships between the recovery rate and the preoperative radiographic factors, and between TK and other radiographic factors. A score of 0.0–0.2 represents “very weak or no correlation”; 0.2–0.4, “weak correlation”; 0.4–0.6, “moderate correlation”; 0.6–0.8, “strong correlation”; and 0.8–1.0, “very strong correlation. The associations between baseline clinical and radiographic variables with recovery rate were investigated with a multiple linear regression model. First, predictors associated with the dependent variable at a analysis . Second,p-value > 0.05 were removed. For all statistical analyses, JMP version 12  was used, and a p-value of < 0.05 was considered statistically significant.Predictors with a A total of 37 patients with 1 year of postoperative follow-up were included in this study. The baseline characteristics of the patients are shown in Table Table ρ = − 0.52, p = 0.001) and TPA ; however, no correlations were observed between the recovery rate and other radiographic parameters. Based on the Spearman correlation coefficients, SVA had a moderate correlation with the recovery rate, and TPA had a weak correlation with the recovery rate.We then investigated whether the recovery rate correlated with the preoperative factors Table . The resp = 0.049, 0.004 and 0.03).The association between the baseline variables and the recovery rate was investigated in a univariate regression model Table . The unip = 0.01), PCS , and SVA   Tables .Table 5Mρ = 0.40, p = 0.02), C7 slope , and LL . Based on the Spearman correlation coefficients, C2-7 SVA and LL had weak correlations with TK, and C7 slope had a strong correlation with TK.Lastly, we examined the correlation between TK and other radiographic parameters. The results showed that TK correlated with the C2-7SVA (This study investigated the predictors of recovery rate after ADF. C-JOA, EQ-5D, NDI, and PCS scores were improved postoperatively. The recovery rate was significantly correlated with the SVA and TPA. Univariate regression analyses showed that the age, SVA, and PCS scores were significantly associated with the recovery rate. Lastly, stepwise multiple regression analysis showed that the independent predictors of recovery rate after ADF were identified as TK, PCS, and SVA. To the best of our knowledge, this study is the first to investigate the predictive value of global spinal parameters and PCS score for recovery rate after ADF.The clinical outcomes of ADF were limited by various factors, such as duration of symptoms, age, BMI, and preoperative MRI spinal cord signal changes , 25. RegWe also found that the independent radiographic predictors of recovery rate after ADF were identified as TK and SVA. These results indicate that patients with higher TK and lower SVA were more likely to improve their C-JOA scores after ADF. A recently established concept is that good SVA can be predicted by pelvic incidence, LL, and TK . In geneWe also found that the independent predictor of recovery rate after ADF was identified as PCS score. A cohort study that investigated the surgical results of cervical spondylotic myelopathy showed that the recovery rate improvements correlated with the physical component domains of SF-36 , althougThere are some limitations in this study. First, although our study is based on prospectively collected data, the primary limitation of this study is the retrospective design. Second, sample size may have been small. Although there is no consensus on the appropriate sample size for the multiple regression analysis, in some studies, 10 events per variable is considered reasonable in a regression analysis , 33. BasWe found that the recovery rate following ADF was negatively correlated with the preoperative SVA and TPA. A preoperatively higher SVA, lower TK, and lower PCS score were independent predictors for poor recovery after ADF. Therefore, patients who undergo ADF with these predictors might be cautioned about poor recovery and be required to provide adequate informed consent.


In May 2012 the US Preventive Task Force issued a ‘D’ recommendation against routine PSA-based early detection of prostate cancer. This recommendation was implemented progressively in our health system. The aim of this study is to define its impact on prostate cancer staging at a tertiary care institution.A retrospective analysis was performed from 2012 until 2015 at a single center. We analyzed the total number of biopsies performed per year and the positive biopsy rate. For those patients with positive biopsies we recorded diagnostic PSA, clinical stage, ISUP grade group, nodal involvement and metastatic status at diagnosis.p < 0.05). No change in median PSA was noticed (p = 0.627). The biopsies detected higher ISUP grades (p = 0.000). In addition, newly diagnosed prostate cancer presented a higher clinical stage (p = 0.005), higher metastatic rates (p = 0.03) and a tendency to higher lymph node involvement although not statistically significant (p = 0.09).A total of 1686 biopsies were analyzed. The positive biopsy rate increased from 25% in 2012 to 40% in 2015 (After the 2012 recommendation, patients presented a higher probability of a prostate cancer diagnosis, with a more adverse ISUP group, clinical stage and metastatic disease.These results should be taken into consideration to implement a risk adapted strategy for prostate cancer screening. Prostate cancer is the second most commonly diagnosed and the most prevalent cancer among males, with 358,989 deaths worldwide during 2018 . Howeverp = 0.00]) at 11 years follow up [In May 2012 the US Preventive Task Force (USPTF) issued a ‘D’ recommendation for routine PSA-based early detection of prostate cancer, stating that it should not be offered in the general U.S. population, regardless of age . This reollow up . The aimp value < 0.05. Analysis was performed with SPSS 23.0 version.After obtaining the institutional ethics committee’s approval, we conducted a retrospective review of all patients who underwent prostate needle biopsies (PNB) at a single tertiary-care institution between January 2012 and December 2015. Patients were excluded from the analysis if they had been previously diagnosed with prostate cancer or had prostatic intraepithelial neoplasia (PIN) or atypical small acinar proliferation (ASAP) in the absence of any prostatic adenocarcinoma. We analyzed the total number of biopsies performed per year and the positive biopsy rate. For those patients with positive biopsies we recorded diagnostic PSA, digital rectal examination (DRE), ISUP grade group, nodal involvement and metastatic status. The Chi square test of independence was used to compare positive biopsy rates, Mann–Whitney U test was used to compare prebiopsy PSA and Chi square Mantel–Haenszel test (linear by linear) for temporal tendency in the rest of the variables. Statistical significance was set at a p < 0.0001).During the period studied, 1686 prostatic needle biopsies were performed. An overall 45% reduction was observed in the number of biopsies performed between the first and last year studied. Table The clinical presentation based on digital rectal examination (DRE), ISUP grade and distant metastasis were significantly worse in the later years studied. The lymph node involvement showed a non-statistically significant increase and the PSA value at diagnosis did not show any difference throughout the study period.p = 0.005) as shown in Fig. p = < 0.05).The proportion of clinical stage T1 and ≥ T3 were 63.3% and 8.0% in 2012. 62.8% and 11.5% in 2013. 58.6% and 14.1% in 2014 51.6% and 18.8% in 2015 respectively (p = 0.024) with a proportion of 8.8%. 11.2%. 9.3% and 18.6% each year .Significant differences were observed in distant metastasis at diagnosis (linear by linear chi square test p = 0.627).Surprisingly, the median PSA was 9.0 ng/dl ICR 6.1–14.3) for 2012; 9.5 ng/dl (ICR 6.3–23) for 2013; 8.3 ng/dl (ICR 5.9–17) for 2014 and 8.3 ng/dl (ICR 6.01–21) for 2015  and with D’Amico high risk scores (p = 0.036) after the USPTF recommendation [We observed a significant increase in local tumoral aggressiveness, mostly because of an increase in the clinical staging cT2-4, and an increase in ISUP 4 and 5. In their study, Banjeri et al., reported similar findings, with higher clinical stage  [p < 0.05) [p < 0.01) [We identified a significant increase in metastatic prostate cancer at the time of initial diagnosis, which, in our opinion, is the most important negative consequence of the implementation of non-screening recommendations. Our data supports the results described by other authors showing an increased incidence of metastatic prostate cancer at time of diagnosis. Bernstein et al., analyzing the SEER database, reported that in men ≥ 75 years old, the diagnosis of distant metastases increased in 2012 compared with 2011  . Using t < 0.05) . In a po < 0.05) . Interes < 0.05) . This is < 0.01) .p = 0.001) and PCa mortality, with a lower number of men needing to be invited for screening and diagnosis to prevent prostate cancer death (570 and 18 respectively) [Updated results from the ERSPC trial confirm the risk reduction of developing metastasis  , 38. It ctively) . Differectively) .Although our study provides important information about the impact of the 2012 recommendation on PSA screening in our population, we are aware of its limitations. In addition to its retrospective design and inherent biases, as a tertiary center with different associated centers we do not know the full level of penetration of the recommendation and the exact time of adoption in primary health centers. In addition, there could be a selection bias as the patients included in the study were not segregated as to whether they are on a screening or non-screening protocol for PNB.After the recommendation against PSA screening, the diagnostic profile of prostate cancer has changed in our tertiary care institution, with prostate cancer being diagnosed at a higher clinical stage, with increased histological aggressiveness and increased risk of metastatic disease. Such findings should favor a PSA- based screening policy for early detection of PCa. Advancing in this direction, different urological societies are working to implement prostate cancer screening for the general population.


Serum amyloid A (SAA), interleukin‐6 (IL‐6) and neutrophil‐to‐lymphocyte ratio (NLR) play critical roles in inflammation and are used in clinical laboratories as indicators of inflammation‐related diseases. We aimed to provide potential laboratory basis for auxiliary distinguishing coronavirus disease (COVID‐19) by monitoring above indicators.A total of 84 patients with confirmed COVID‐19 were enrolled in the study. Baseline characteristics and laboratory results were collected and analyzed. Receiver operating characteristic (ROC) curve analysis was used to combined detection of SAA and IL‐6 in patients with COVID‐19, and independent risk factors for severity of COVID‐19 were assessed by using binary logistic regression.P < .001), and compared with nonsevere patients, three indicators of severe patients were significantly elevated. Besides, combined detection of SAA and IL‐6 better separates healthy people from patients with COVID‐19 than detection of SAA or IL‐6 alone. In addition, elevated SAA, IL‐6, and NLR can be used as independent variables for predicting the severity of patients with COVID‐19.The main clinical symptoms of patients with COVID‐19 were fever (98.8%), fatigue (61.9%), and dry cough (58.3%). SAA, IL‐6, and NLR were significantly higher in patients with COVID‐19 (all Serum amyloid A and IL‐6 could be used as addition parameters to helping the distinguish of patients with COVID‐19 from healthy people, and can provide potential basis for separating patients with nonsevere and severe clinical signs. Comparison of laboratory indicators among patients with COVID‐19 ARDSacute respiratory distress syndromeCOVID‐19coronavirus diseaseCRPC‐reactive proteinIQRinterquartile rangeMERSMiddle East respiratory syndromeNLRneutrophil‐to‐lymphocyte ratioORodds ratioPLRplatelet‐to‐lymphocyte ratioROCreceiver operating characteristicSAAserum amyloid A; IL‐6: interleukin‐6SARSsevere acute respiratory syndromeSARS‐CoV‐2severe acute respiratory syndrome coronavirus 2WHOWorld Health Organization1http://2019ncov.chinacdc.cn/nCoV/), as of March 11, 2020, at 10:00, China had cumulatively diagnosed approximately 80 955 patients with coronavirus disease (COVID‐19), and other countries have accumulated approximately 36,900 cases. Basic, clinical, and epidemiological research on COVID‐19 continues to be reported.Coronaviruses mainly cause respiratory infections and some strains, such as severe acute respiratory syndrome (SARS) and Middle East respiratory syndrome (MERS), are associated with high infectivity and mortality, and thus are harmful to public health.Existing reports show that approximately 84% of patients were categorized as nonsevere cases based on clinical symptoms, such as fever, cough, myalgia, or fatigue.22.1A total of 84 patients confirmed COVID‐19 admitted to the Second People's Hospital of Lianyungan and Wuxi Fifth People's Hospital from January 23, 2020, to February 29, 2020, were enrolled. The diagnosis of COVID‐19 was based on WHO interim guidanceAccording to the novel coronavirus pneumonia diagnosis and treatment plan ,2.2The white blood cell count, neutrophil count, lymphocyte count, red blood cell count, platelet count, and hemoglobin and C‐reactive protein (CRP) levels were determined with a BC‐5390 automatic hematology analyzer and associated reagents . SAA was determined with commercially available assay kits  and an automatic biochemical analyzer  by using the method of latex immunoturbidimetry. IL‐6 was measured with a commercially available assay kit  and an Access 2 automatic immune analysis system  by using the method of enzyme‐linked immunochemiluminescence. All equipment was maintained and calibrated according to the requirements. Internal quality control and external quality assessment were performed.2.32 anticoagulated venous blood was collected on admission for detection of the white blood cell count, neutrophil count, lymphocyte count, red blood cell count, platelet count, and hemoglobin and CRP levels. Meanwhile, 5 mL of venous blood (with a separation tube) was collected for the detection of SAA and IL‐6. The latter specimens were centrifuged at 1200 × g for 10 minutes, and the detection of all analytes was completed within 4 hours.From all subjects, 2 mL of EDTA K2.4P value <.05 was considered statistically significant.Data were analyzed in IBM SPSS Statistics 21.0, and data are expressed as median with interquartile range (IQR). The Mann‐Whitney test was used to compare two independent groups. The Kruskal‐Wallis test was used to compare multiple groups, and Dunnett's test was used for pairwise comparisons. Categorical variables were analyzed with chi‐square test. The combined detection of laboratory indicators was analyzed with the receiver operating characteristic (ROC) curve, and risk factors were evaluated with binary logistic regression. A 33.1P > .05). The median blood pressure (systolic pressure and diastolic pressure) in the two groups was significantly different (P < .05). Both systolic and diastolic blood pressure were significantly higher in the severe group than in the nonsevere group. Among the 84 patients, 83 (98.8%) had fever, 52 (61.9%) had fatigue, 49 (58.3%) had dry cough, 31 (36.9%) had myalgia, 26 (31.0%) had headache, 16 (19.0%) had expectoration, 14 (16.7%) had pharyngalgia, 10 (11.9%) had diarrhea, and eight (9.5%) had nausea or vomiting; however, there was no statistical difference between the nonsevere group and severe group (P > .05). Of the 84 patients with COVID‐19, 33 (39.3%) had anorexia, and 23 (27.4%) had dyspnea; there was a statistical difference between the two groups (P < .05). Among the 84 patients, five (6.0%) had diabetes, two (2.4%) had cardiovascular disease, four (4.8%) had cerebrovascular disease, three (3.6%) hadchronic kidney disease, five (6.0%) had chronic obstructive pulmonary disease, and one (1.2%) patient had cancer; however, there was no statistical difference between the nonsevere group and severe group (P > .05) , and 25 were placed in the severe group  on admission. There was no statistical difference in the sex ratio, age, body mass index (BMI), and smoking status between the nonsevere group and severe group (3.2P < .05). Moreover, the neutrophil‐to‐lymphocyte ratio (NLR), SAA, and IL‐6 were significantly higher in patients with COVID‐19 than in the healthy control group. In addition, the NLR, SAA, and IL‐6 in patients were significantly higher in the severe group than in the nonsevere group (P < .05). Compared with healthy controls and nonsevere group, platelet‐to lymphocyte‐ratio (PLR) was significantly increased in severe group (P < .05). However, the white blood cell count, neutrophil count, red blood cell count, platelet count, and hemoglobin were not statistically different between healthy controls and patients with COVID‐19 , but there was no statistical difference between the nonsevere group and the severe group. The levels of CRP were significantly higher in patients with severe disease than healthy controls and patients with nonsevere disease . Therefore, our study indicated that SAA, IL‐6, and NLR can be used as independent variables for predicting the severity of patients with COVID‐19 (Table All patients with COVID‐19 (nonsevere and severe group) were considered as dependent variables, and age, body mass index (BMI), smoking status, coexisting disorder, white blood cell count, neutrophil count, lymphocyte count, NLR, PLR, CRP, SAA, and IL‐6 were considered the independent variables. Binary logistic regression analysis showed that one unit increase in SAA, IL‐6, and NLR, the risk of nonsevere patients with COVID‐19 becoming severe patients increased significantly ,We analyzed the results of laboratory indicators and found that the white blood cell count and neutrophil count of the patients with COVID‐19 did not significantly differ, but the lymphocyte count was significantly lower than that in the healthy group. Moreover, the NLR of patients with COVID‐19 was significantly higher than that of the healthy group, and it was higher in patients with severe disease. A study conducted by Liu et alThrough the combined detection of SAA and IL‐6 in patients, we also found that combined detection of the two can significantly improve the distinguishing of patients with COVID‐19 from healthy people. In addition, we further analyzed the independent variables for predicting the severity of COVID‐19. And we found that patients with higher levels of SAA, IL‐6, and NLR have greater risk to develop severe disease. Chen et alThere are some limitations to this study. Firstly, because only 84 patients with COVID‐19 were included, our conclusions must be further verified in more participants. Secondly, as patients come from different hospitals or regions, we have some difficulties in obtaining patient's data, which leads to incomplete acquire to some data, such as average time between infection and occurence of symptoms, the time from admission to the hospital and developing severe type, and part of laboratory indicators. Thirdly, this was a cross‐sectional study, and we were unable to analyze the changes in laboratory indicators in patients in the treatment and prognosis stages; consequently, we lack corresponding information on the prognosis of the disease. Therefore, the changes in various indicators must be analyzed after treatment of patients to provide comprehensive reference values for disease prognosis.In conclusion, SAA and IL‐6 could be used as addition parameters to helping the distinguish of patients with COVID‐19 from healthy people, and can provide potential basis for separating patients with nonsevere and severe clinical signs.QL and FMY conceived and designed the experiments. QL, YPD, and XW performed the experiments. MMF and WL analyzed the data. QL, YPD, and FMY wrote the article.