Despite the proportion of elderly breast cancer patients has been consistently increasing, the optimal treatment modalities for this population have not been well explored. We summarized the treatment outcomes of these patients in our hospital.Older patients with early breast cancer were identified from the Breast Cancer Information Management System at West China Hospital, Sichuan University (2000–2019). We compared tumor characteristics and treatment outcomes between the older group (65–74 years old) and the elderly group (≥75 years old). The Kaplan-Meier and Cox regression analysis were conducted to determine significant prognostic factors.P = 0.035). Subgroup analysis revealed that patients with grade III disease best benefited from adjuvant chemotherapy. PMRT offered a significant improvement in local disease control, but not in OS. Furthermore, endocrine therapy improved the OS of HR-positive patients , especially for cases aged 65–74 years. Also, receipt of trastuzumab in HER2-positive patients was associated with better OS .In total, 1094 patients were included. The median follow-up time for this cohort was 59 months. The majority of patients underwent surgery and benefited from surgical treatment. Elderly group patients were less likely to receive adjuvant chemotherapy or postmastectomy radiotherapy (PMRT) compared to the older group. However, adjuvant chemotherapy was associated with improved overall survival (OS) (hazard ratio [HR] 0.521, 95% confidence interval [CI] 0.284–0.955, Our findings suggest that surgery, adjuvant chemotherapy, endocrine and targeted therapy are associated with improved OS in older breast cancer patients. Moreover, clinicopathological characteristics should be comprehensively considered when making treatment decisions for these patients.The online version contains supplementary material available at 10.1186/s12885-021-07947-w. Breast cancer is the most common malignancy in women worldwide, and the incidence rates have been rising for most countries in transition over the last decades . From 20However, despite the increasing proportion of elderly patients with breast cancer, data on treatments and outcomes for this patient population are limited, partly because elderly patients have been under-represented in most breast cancer treatment trials . PreviouTherefore, this study aimed to investigate the tumor characteristics and the associations between treatment modalities and survival in a large population-based cohort of patients over 65 years of age with early breast cancer.n = 1145). Patients with synchronous distant metastases (n = 44) or those lacking essential registry data (n = 7) were excluded. In total, 1094 consecutive female patients were included in this cohort. Ethical approval for this study was provided by the Clinical Test and Biomedical Ethics Committee , West China Hospital, Sichuan University. And all methods were performed in accordance with the Declaration of Helsinki. Informed consent was obtained from all patients at the time of initial diagnosis.Breast cancer information management system (BCIMS) of West China Hospital, Sichuan University, contains over 16,000 breast cancer patient cases dating back to 1989 and prospectively records demographic and clinicopathologic characteristics, medical history, diagnosis, laboratory results and treatments, as described previously . Female Demographic, clinicopathologic, and treatment information were derived from the BCIMS. Demographic records included age at diagnosis and body mass index (BMI). The BMI cutoffs are based on the standards recommended in the Guidelines for Prevention and Control of Overweight and Obesity in Chinese Adults . ClinicoThe follow-up was conducted every 4 months in the first 3 years after diagnosis, every 6 to 12 months in the fourth and fifth years, and annually after 5 years. Follow-up was operated by office visit, telephone, email, or postal contact, as described in our previous report . Lost toOverall survival (OS) was calculated as the time from the date of diagnosis to the date of death from any cause. Disease free survival (DFS) was defined as the time from diagnosis to the first event . Locoregional recurrence (LRR) was defined as recurrence in the ipsilateral chest wall, supraclavicular and infraclavicular areas, axilla, and internal mammary region. Locoregional recurrence-free survival (LRFS) was counted from surgery date to the date of LRR as the first event . PatientP values less than 0.05 were considered to be statistically significant. All statistical analyses were performed using IBM SPSS statistics software (version 23.0) and R software .Data are presented as medians or proportions. Descriptive statistics were used to analyze the clinical and demographic characteristics of the population. According to the nature of the variable, the Chi-square test, Fisher’s exact test or Student’s t-test was used to compare the distribution of baseline characteristics among age groups. Survival curves were estimated using the Kaplan-Meier method, and log-rank tests were performed for comparison of survival outcomes. Cox proportional hazards regression models were used to determine the significant prognostic factors of survival and expressed as hazard ratio (HR) with 95% confidence interval (CI). Missing data were processed using multiple imputation. All statistical tests were 2-tailed and A total of 1094 women with early-stage breast cancer were included in the analysis. The median age at diagnosis of breast cancer was 68 years (range 65–94 years) for the whole patients. The demographic and tumor characteristics of patients are summarized in Table P <  0.001). No significant differences were found between two age groups in terms of tumor histological type, clinical stage, T stage, N stage, HR status, HER2 status, molecular subtype, Ki-67 level and tumor grade.All patients were categorized into two subgroups based on their age at diagnosis: the older group (65–74 years old) versus the elderly group (≥ 75 years old). The total number of older group patients was 930 (85.0%), and that of elderly group was 164 (15.0%). The elderly group patients were more likely to be underweight when compared with the older group (P <  0.001). However, recurrence and metastasis rate was not different between the two groups .The median follow-up time for this cohort was 59 months. Death events differed between the two groups, 102 (11.0%) cases died in the older group, and 41 (25.0%) cases died in the elderly group during the follow-up period (P <  0.05). Whereas, in the multivariate analysis, only age older than 75 years , underweight , stage III  and high Ki-67 level  were independently associated with poor survival. Furthermore, we performed multivariate Cox regression analysis for DFS and found only underweight and stage III were independent risk factors of DFS for the entire population . Of those who underwent surgery, 963 patients (92.3%) received modified radical mastectomy, while 54 patients (5.2%) received radical mastectomy, very few patients  were treated with breast conserving surgery or palliative surgery.The majority of patients  in this cohort underwent surgery  received adjuvant chemotherapy, but elderly group patients were less likely to receive it when compared with those in the older group . Among these patients, only 90 (12.3%) cases received neoadjuvant chemotherapy, including 83 (13.2%) cases in the older group and 7 (6.7%) in the elderly group. Neoadjuvant chemotherapy was not associated with survival in multivariate analysis . Of these, only 134 (29.6%) patients received postmastectomy radiotherapy, and the older group was more likely to receive adjuvant radiotherapy compared with the elderly group , approximately one-third of patients received trastuzumab treatment. And no difference was observed in the proportion of patients receiving trastuzumab between two groups  was shown to reduce the risks of both recurrence and breast cancer mortality in all patients with node-positive breast cancer considered together . In addiAmong HR-positive older patients, the proportion of the elderly group who accepted endocrine therapy was similar to that of older group in our current study. In contrast, several studies reported that women aged > 75 years were less likely to receive endocrine therapy compared with younger patients , 42. As In addition, we described the use and outcome of trastuzumab treatment among older women with HER2-positive breast cancer. Only one third of HER2+ patients received trastuzumab treatment in this cohort, this finding was in line with recent data, they observed that the low utilization of adjuvant trastuzumab for older patients with HER2-positive disease . HoweverThis study had some limitations. Our study did not contain information on treatment-related complications, functional status, patient preference and quality of life, which may affect choice of treatment and survival outcomes. Our findings were based on a large cohort from a single center, thus need further verification in independent data sets.In this large cohort, we found that surgical treatment remains a routine and curative option for older patients with early breast cancer. In the adjuvant setting, use of chemotherapy in older patients at high risk of recurrence (grade III or higher stage) was associated with improved survival. And endocrine therapy in HR-positive breast cancer patients and trastuzumab in HER2-positive breast cancer patients may be associated with reduced risk of mortality. We didn’t find neoadjuvant chemotherapy or postmastectomy radiotherapy conferred significant OS or DFS benefit on older breast cancer patients. Moreover, prognostic clinicopathological characteristics should be comprehensively considered when making treatment decisions for those patients.Additional file 1: Table S1. Univariate and multivariate analysis of disease free survival in all breast cancer patients. Table S2. Multivariate analysis of the effect of treatment on disease free survival in different population.Additional file 2: Fig. S1. Kaplan–Meier estimates of disease free survival by treatment modalities. Fig. S2. Subgroup analyses of impact of treatment modalities on disease free survival.


Obesity is a disease characterized by much fat accumulation and abnormal distribution, which was related to cardiovascular diseases, diabetes mellitus (DM) and muscular skeletal diseases. The aim of this study was to evaluate the usefulness of appendicular skeletal muscle mass to total body fat ratio (ASM/TBF) in screening for the risk of obesity in elderly people.2) who underwent baseline and an average around 2.2-year follow-up health check-up examinations.A prospective study was carried out with 446 participants (non-obese elderly people with body mass index (BMI) < 28 kg/mβ = − 1.147 (− 1.463—-0.831) for men and − 4.727 (− 5.761—-3.692) for women). The cut-off points of baseline ASM/TBF in elderly people for obesity were 1.24 in men and 0.90 in women which were identified by Classification and Regression Tree (CART). Logistic regression showed that both men and women with decreased ASM/TBF had higher risks of obesity over the follow-up period (Relative Risk (RRs) = 5.664 (1.879–17.074) for men and 34.856 (3.930–309.153) for women).The mean age at baseline was 63.6 years. The incidence of new obesity was 5.4% during follow-up. Linear regression demonstrated that baseline ASM/TBFs were negatively correlated with follow-up BMIs in both men and women (Elderly people with a low ASM/TBF had a higher risk of new obesity, which suggested that ASM/TBF should be considered in obesity management in the elderly. Obesity is a disease characterized by much fat accumulation and abnormal distribution, which is related to cardiovascular diseases, diabetes mellitus (DM) and muscular skeletal diseases . Severe The close relationship between appendicular skeletal muscle mass (ASM) / total body fat (TBF) and sarcopenia obesity, which was defined by top TBF quartile, has been well recognized in previous studies . HoweverASM and TBF, represented muscle and fat components and measured by Bioelectric impedance (BIA) technique, were shown after safe current passed through the human body , 8. PrevAlthough ASM/TBF has been demonstrated previously , little Elderly people aged over 60 years who underwent a routine health check-up in Chinese PLA General Hospital from February 2013 to February 2014 were enrolled in this study. The inclusion criteria were elderly people aged over 60 who accepted BIA and anthropometric measurements. Participants were excluded for any of those cases: (a) extremities diseases; (b) previous obesity; (c) diagnosed metastatic advanced tumor or malignant tumor discovery less than 5 years; (d) secondary obesity caused by several diseases, such as Cushing syndrome. Five hundred and forty-five participants were recruited at baseline. Visits were performed at baseline and the next before February 2016. The follow-up time was between 0.6–3.8 years. Finally, a total of 446 participants completed the average around 2.2 years follow-up. The missing rate was 18.2%. All participants were given written informed consents and the study protocol was approved by the Institutional Review Board of Chinese PLA General Hospital.Past history  and personal history  were collected by trained healthcare providers. The method of BIA measurement can be seen from our teams’ previous works , 11. Oth2  and count (percentage), respectively. Classification and Regression Tree (CART) Models, measured in terms of true classification rate of the classifier, were built through a process known as binary recursive partitioning, which is an iterative process that splits the data into partitions or branches, and then continues splitting each partition into smaller groups as the method moves up each branch. The purpose of the analyses via tree-building algorithms was to determine a set of if-then logical (split) conditions that permit accurate prediction or classification of cases. Cut-off values of baseline ASM/TBF for obesity were identified by CART. Moreover, logistic regression was used to examine P < 0.001 in women and P = 0.119 in men, Table The average age at baseline was 63.6 years. The minimum age was 60 and the maximum age was 81. The incidence of new obesity was 5.4% during follow-up. Subjects with new obesity had significantly lower baseline ASM/TBF than subjects without new obesity  in both men and women  in model 6 were 1.879–17.074 and 3.930–309.153, separately)  (Table There were 58 participants (13.0%) with decreased ASM/TBF both at baseline and follow-up. Logistic regression showed that participants with decreased ASM/TBF both at baseline and follow-up demonstrated higher risk of obesity (This study reported for the first time that the usefulness of ASM/TBF in predicting for the risk of obesity, which was diagnosed by BMI, in elderly people.The relationship between fat related indicators and obesity has been widely discussed in previous studies . As BMI The opinions of obesity were controversial in the elderly. One study involved elderly men in Australia, defined obesity as more than 30% body fat, showed that simple obesity was a protective factor for frailty . But anoBesides, there was no agreement about cut-off points for ASM/TBF to predict obesity. Jean Woo et al.  has repoFrom the view of histology and embryology, interestingly, skeletal muscle cells shared a common developmental origin with brown adipocytes, which broke down lipids to generate heat, thus reducing obesity . The potEven though it was a longitudinal study, the work had inevitable limitations. Firstly, as a single center study, its biases were unavoidable. For example, because of Berkson bias, a few number of elderly people aged over 80 was recruited in this study. So whether ASM/TBF was an universal and representative assessing indicator to older people needs further exploration. Secondly, visceral fat mass, which was a potential confounding variable, was not demonstrated in this study, therefore a large and longitudinal cohort is warranted to further demonstrate the relationship between visceral fat mass related indicators and obesity. Thirdly, because the new obese population is so small, the results from logistic regression may not be very reliable, which needs amplified sample size to further verify the predictive value of ASM/TBF in obesity.Elderly people with decreased ASM/TBF had higher risk of new obesity, which suggested that ASM/TBF should be considered in obesity management in elderly people.


Automatic segmentations achieved a median dice similarity coefficient (DSC) of 0.80 compared to the reference standard. Aneurysm location  and bleeding severity  did not impede detection sensitivity or segmentation performance. For aneurysms > 100 mm3 (mean diameter of ~ 6 mm), a sensitivity of 96% with DSC of 0.87 and FPs/scan of 0.14 were obtained. In the present study, we demonstrate that the proposed DLM detects and segments aneurysms > 30 mm3 in patients with aSAH with high sensitivity independent of cerebral circulation and bleeding severity while producing FP findings of less than one per scan. Hence, the DLM can potentially assist treating physicians in aSAH by providing automated detection and segmentations of aneurysms.In aneurysmal subarachnoid hemorrhage (aSAH), accurate diagnosis of aneurysm is essential for subsequent treatment to prevent rebleeding. However, aneurysm detection proves to be challenging and time-consuming. The purpose of this study was to develop and evaluate a deep learning model (DLM) to automatically detect and segment aneurysms in patients with aSAH on computed tomography angiography. In this retrospective single-center study, three different DLMs were trained on 68 patients with 79 aneurysms treated for aSAH (2016–2017) using five-fold-cross-validation. Their outputs were combined to a single DLM via ensemble-learning. The DLM was evaluated on an independent test set consisting of 185 patients with 215 aneurysms (2010–2015). Independent manual segmentations of aneurysms in a 3D voxel-wise manner by two readers  provided the reference standard. For aneurysms > 30 mm In patients with non-traumatic SAH, accurate and reliable diagnosis of aneurysm is essential for subsequent treatment to prevent re-bleeding and further neurological deterioration4. Usually, CT-angiography (CTA) is performed immediately upon radiological proof of SAH with sensitivity rates for detection of aneurysms ranging between 85–98% compared to digital subtraction angiography (DSA), which is considered as the gold standard for aneurysm imaging6. Due to advances in imaging and diagnostic quality over the last decades as well as its non-invasiveness, CTA has the potential to replace DSA in pre-treatment assessment of aSAH in selected patients8. Timely aneurysm occlusion by endovascular or surgical means represents a key concept in modern aSAH management with selection of most suitable treatment depending on various factors such as aneurysm localization, size, and shape9.Aneurysmal subarachnoid hemorrhage (aSAH) is caused by spontaneous rupture of an intracranial aneurysm and represents a severe neurological condition with mortality ranging between 8 and 67%12. Aneurysm detection on CTA, especially for smaller ones, proves to be challenging and misdiagnosis of aSAH can be associated with a poor clinical outcome15. Hence, the development of a deep learning model (DLM) to automatically detect and segment intracranial aneurysms would be of valuable assistance to the radiologist. This is of particular interest due to the growing workload and consequent fatigue of radiologists, which correlates with increased risk to miss relevant findings16.With the introduction and advancements of convolutional neural networks (CNN) over the last decade, deep learning algorithms have shown great potential in performing diagnostic and analyzing tasks on medical imaging in different subspecialties10 and time-of-flight magnetic resonance angiography (TOF-MRA)18. However, these studies focused on unruptured intracranial aneurysms (UIAs) and did not include patients with aSAH. Hence, it remains unclear, how DLM algorithms perform on patients with acutely ruptured intracranial aneurysms (RIAs) and whether the extent of hemorrhage impedes detection sensitivity.Previous studies have proposed several approaches for (semi-) automated detection of intracranial aneurysms on CTAThe objective of this study was to develop and validate a DLM for automatic detection and segmentation of aneurysms on CTA in aSAH. Furthermore, we evaluated the performance of the algorithm with regard to aneurysm size, location, and bleeding severity using an independent test set of patients with aSAH., v3.0, Philips Healthcare, Best, the Netherlands).All consecutive patients treated for aSAH at our tertiary-care university hospital setting between January 2010 and December 2017 were reviewed and served as our dataset (n = 340). Exclusion criteria were: (1) unavailable CTA scans, (2) no distinct aneurysm finding in CTA, (3) severe motion artifacts on CTA, (4) insufficient contrast of CTA, and (5) previously treated aneurysms. There were no exclusions due to aneurysm size. Consequently, 87 patients were excluded, resulting in 253 patients  with 294 aneurysms for analysis. All included scans between 2016 and 2017 (n = 68 patients/79 aneurysms) served as the training dataset, whereas scans before 2016 were allocated to the independent test set (n = 185 patients/215 aneurysms). Figure Philips Healthcare, Best, the Netherlands). On all of these scanners, our institutional standard clinical protocol for head and neck (n = 222) or head (n = 29) CTA was used with slice thickness ranging between 0.62 to 1.25 mm. Two CTA datasets of the head from referring hospitals were included. These were acquired using Siemens Somatom Definition AS  and Toshiba Aquilion 64 . (Pre-) interventional DSA was performed on a biplane angiography system . Two-hundred-thirty-four patients received (pre-) interventional DSA.Included examinations were performed on different multidetector-CTs, namely iCT (n = 229), Brilliance 64 (n = 8), and Brilliance 16 (n = 14) . Further, they collectively reviewed non-enhanced CT scans to determine respective Fisher grade of aSAH in consensus. Reference standard for aneurysm segmentations were provided by the above-mentioned neurosurgeon and radiologist, who performed semi-automatic 3D voxel-wise segmentations of aneurysms on ISD in consensus.Wellcome Trust Centre for Neuroimaging)19. Second, a multi-scale vessel enhancement filter was applied to the brain masked images to enhance the arteries from the background of CTA scans20. In this context, two vessel-enhanced images were computed; one with scale 0.5–5 voxels and the other with scale 5–15 voxels. The two vessel-enhanced images together would help to distinguish between blood vessels and aneurysms. Third, image standardization was performed by resampling to isotropic resolution of 0.5 × 0.5 × 0.5 mm and intensity normalized. The original CTA image was normalized between 5–95% of its intensity values and the vessel enhanced images were Z-score normalized. The fully automatic image pre-processing workflow is shown in Fig. To enable DLM-based automatic aneurysm segmentation workflow, a number of preprocessing steps were involved. First, a brain extraction algorithm was developed to compute the brain mask, using Statistical Parametric Mapping software package version 8  consisting of a deep 3D CNN architecture with 2 identical pathways that apply different image resolutions to capture contextual information21. 3D image-segments centered at the same image location provide inputs to the two pathways. However, for the second pathway, the image is down-sampled to a third of its original size. The model comprises of 11 layers with kernels of size 33. The model also consists of residual connections for layers 4, 6, 8, and 10 whereas layers 9 and 10 are fully connected.In this study, a 3D CNN was used. The CNN is based on DeepMedic  For DLM-Vess, the input to the CNN is multi-channel comprising of the CTA source images and the two vessel enhanced images. The size of the extracted image-segments was set to 253 voxels. (III) For DLM-LDim, CTA source images represent the input, but the size of the extracted image-segments was increased from 253 to 453 voxels.(I) For DLM-Orig, the input to the CNN is a single channel original CTA image with the size of extracted image-segments for training being set to 25To increase the amount of training samples for all three models, image augmentation was employed by flipping the images along their axes. Training batch size was set to 15, batch normalization was applied and parametric rectified linear unit was used as the activation function, Dice similarity coefficient (DSC) was provided as the loss function and the number of training epochs was set to 30.22, we created an ensemble model by combining the outputs of the three separate DLMs. We refer to this combination strategy as DLM-Ens.The three DLMs were trained on the training-set by a five-fold-cross-validation approach using an 80–20% training-validation split without overlapping data. Similar to the work of Kamnitsas et al.23. Subsequently, STAPLE outputs from the three DLMs were passed to DLM-Ens to produce final aneurysm segmentation prediction.During inference, the trained DLMs  were applied to the test dataset. Each trained DLM consisted of five individual sub-models from the five-fold-cross-validation training approach. Outputs from these five sub-models were fused together using simultaneous truth and performance level estimation (STAPLE)IBM Armonk, NY, USA), with P < .05 considered statistically significant. Categorical variables  are presented in form of percentages, frequencies, and medians. Comparison of paired nonparametric variables was performed with Wilcoxon signed-rank tests. Normally distributed variables are given as mean ± standard deviation with comparisons being conducted using Student’s t-test.Statistical analysis was performed with SPSS  was calculated.The local institutional review board approved this retrospective, single-center study  and waived the requirement for written informed patient consent. All methods were conducted in accordance with the relevant guidelines and regulations.Baseline patient and aneurysm characteristics were comparable between the training and validation groups, as outlined in Table In the training set, 79 aneurysms were identified as the reference standard.Using five-fold-cross-validation for each of the three DLMs , an overall sensitivity of 76%, 76%, and 70%, a DSC of 0.54, 0.58, and 0.69, and average false positives (FPs)/scan of 2.10, 2.00, and 0.71 were observed.3 (n = 64) and > 50 mm3 (n = 55), the DLM-Ens achieved higher sensitivity  and higher DSC  with high precision  and decreased number of FPs/scan . In this context, Table In contrast, the DLM-Ens provided a sensitivity of 72% with median DSC of 0.74, precision of 80% and FPs/scan of 0.21. For aneurysms > 30 mm3 were identified as the reference standard on the independent test set.Based on manual 3D segmentations, 215 aneurysms with a mean volume of 145.8 ± 230.3 mm3, thus achieving a significant correlation with manual segmentations . The strongest correlation between manual annotations and automatic DLM segmentations was observed for large aneurysms, while correlation was insignificant (P = .810) for small aneurysms, as detailed in Table For true positive aneurysm findings, the DLM achieved a mean volume of 154.4 ± 236.0 mmVolume correlation plots between manual and automatic segmentations of training and test set using the DLM are presented in Fig. 3 (n = 171), sensitivity was 87%  with FPs/scan of 0.42 (P < .001). Considering aneurysms with volumes of > 50 mm3 (n = 124) and > 100 mm3 (n = 73), corresponding sensitivities of 90%  and 96%  were noted. Table Overall, the DLM achieved a sensitivity of 82%  and FPs/scan of 0.81. For aneurysms > 30 mmPairwise comparison indicated a correlation between detection sensitivity and parent artery location. The sensitivity to detect middle cerebral artery (MCA) aneurysms (88%) was significantly higher than for internal carotid artery (ICA) aneurysms . Likewise, anterior cerebral artery (ACA) aneurysms (84%) and posterior circulation aneurysms (87%) were associated with higher detection sensitivity than ICA aneurysms. However, these differences did not reach statistical significance. When comparing detection rates of anterior  with posterior circulation, no statistical significance was found (P = .07).Detection sensitivity for aneurysms was not significantly impeded by the presence of intracerebral or intraventricular hemorrhage , FP/scan 0.85 (P = .49)).Exemplary images on the test set regarding detected and missed aneurysms as well as FP findings using the DLM are shown in Figs. 3 with high segmentation performance and less than one FP finding per scan.In the present study, we developed and trained a DLM for automatic detection and segmentation of intracranial aneurysms on CTA in patients with aSAH and evaluated its performance on an independent test set. As a major finding of the study, the DLM achieves a detection rate of 87% for aneurysms > 30 mm18 and CTA10, respectively. However, these studies reported FPs/scan of 618 and 1017 or did not report them10, which questions their usefulness in clinical routine. The present study is the first to evaluate the performance of a DLM for automatic detection and segmentation of aneurysms in patients with aSAH. In the current study, the DLM provided a sensitivity of 87% for aneurysms > 30 mm3 (~ mean diameter of 4 mm), which is comparable to aforementioned studies by Ueda et al. (83–95% for UIAs > 3 mm on TOF-MRA)17, Park et al. (95% for UIAs > 3 mm on CTA)10, and Sichtermann et al. (90–93% for UIAs on TOF-MRA)18. For aneurysms > 100 mm3 (~ mean diameter of 6 mm), sensitivity of the DLM increased to almost 100%.Since aneurysms > 7 mm bear the highest risk of rupture26, the proposed DLM enables accurate detection of the clinically most relevant aneurysms in aSAH.Previous studies have investigated DLMs for detection of UIA providing detection rates between 83 and 95% on TOF-MRA3, the proposed DLM yields high segmentation performance  despite the small lesion size. Therefore, the DLM can be of assistance to treating physicians regarding treatment planning, e.g. providing 3D geometrical characterization of aneurysms.Achieving an overall DSC of 0.75 for aneurysm segmentation and of 0.87 for aneurysms > 100 mm18. Concerning detection of larger aneurysms (> 100 mm3), FPs were almost zero per scan. For remaining findings, majority of FPs were found in venous vessels or adjacent to bone and usually not associated with arterial vessels. Therefore, most of them were easily identifiable as an incorrect finding.Using ensemble learning of three different DLMs, an average number of FPs of less than one per scan was observed, being lower than in aforementioned studies27. In future, additional training methods identifying bone geometry might help the DLM to better distinguish bone from adjacent vessels and aneurysms.There was no significant difference between the anterior and posterior circulation regarding aneurysm detection. However, ICA aneurysms showed the lowest detection rates with a significant difference compared to the MCA. This discrepancy is most likely due to presence of calcified plaques, dilatation and elongation of the vessel due to hypertension, complex anatomy of aneurysms, and, in particular, proximity to the skull28.Furthermore, we evaluated whether the presence of aSAH would impede the sensitivity of the DLM. Remarkably, the algorithm performed independently of the Fisher grade with additional presence of parenchymal or intraventricular hemorrhage neither decreasing detection rates nor increasing FPs/scan. Therefore, the DLM provides high feasibility and detection sensitivity even in severe aSAH with potential hydrocephalus or midline shift possibly leading to an overall complex image of the brain14. In this study, we aimed to overcome these limitations by establishing a robust reference standard based on review of CTA scans and reports by three readers as well as availability of DSA in the majority of patients to find aneurysms potentially not been described in the initial CTA report.CTA-based detection of intracranial aneurysms can be time consuming and challenging and shows a large variability among physicians, especially for small aneurysms. They present a highly variable interrater agreement depending on various factors, e.g., localization and subspecialty training, hence resulting in a lower detection rate than larger aneurysms even for experienced clinicians26. With an overall detection rate of 82%, the DLM enables detection rates comparable to a human reader . In this context, the DLM may provide support to physicians that lack concentration due to fatigue or lack of training. This is important in the setting of aSAH if the treating physician detects one aneurysm, preferably the one causing the bleeding, but potentially misses a second aneurysm caused by decreased concentration due to the “satisfaction of search” phenomenon30. Nevertheless, the DLM should be further improved to achieve 100% detection sensitivity, which is required to guarantee patient safety in aSAH.As proof of concept, the results of this study indicate that deep learning is able to provide sufficient detection of aneurysms in aSAH, especially of larger ones, which are bearing the highest risk of ruptureBesides its retrospective design, our study has a few limitations. Although scans from five different CT scanners were included in this study, 91% were acquired using the iCT, hence the true performance of the DLM on CTA images acquired on different scanners besides the iCT is unknown and the evaluation of generalizability is limited. This limitation should be addressed in future studies. Being a single-center study, only two scans were included from referring institutions, therefore a multi-center study investigating the performance of the DLM on other CT scanners using different protocols should be conducted. Further, we did not include previously treated aneurysms; hence, the detection performance in these patients  still needs to be investigated.3 (~ mean diameter of 6 mm). Furthermore, high volumetric correlation to human segmentations and a low number of FPs/scan were obtained. Confounders such as cerebral circulation and bleeding severity did not significantly affect the performance of the DLM. Additional training is required to increase sensitivity for smaller aneurysms; however, the DLM may already be of assistance to treating physicians by providing automated detection of aneurysms in aSAH.In conclusion, we developed a DLM able to provide sufficient detection of aneurysms in aSAH with almost 100% sensitivity for aneurysms > 100 mm


Background/Aims. Hepatorenal syndrome (HRS) is a form of functional renal failure arising in advanced cirrhosis and is characterized by a poor survival rate. Anemia is frequently observed during the clinical course of cirrhosis. Our study aimed to investigate the hematologic findings in patients with cirrhosis to determine the effects of anemia on renal functions in type 2 HRS and if it was a potential aggravating factor. Materials and Methods. This prospective study, in which all consecutive patients with cirrhosis were enrolled, was performed at a tertiary-level hospital  from January 2019 to June 2019. A total of 9 patients with HRS fulfilled the type 2 HRS diagnostic criteria, and 41 patients with cirrhosis without HRS were included. All data regarding patients were obtained from the medical record. Demographic data, routine hemograms, biochemical, and urinary test results were collected. Models of end-stage liver disease (MELD) and Child–Turcotte–Pugh (CTP) scores were calculated. Results. The most common etiology of cirrhosis was viral hepatitis (66%). According to the CTP score, 23 patients were in the CTP-A stage, 13 in the CTP-B stage, and 14 patients were in the CTP-C stage. Patients with type 2 HRS had significantly lower hemoglobin levels compared with non-HRS stable cirrhosis patients. As hemoglobin levels decreased, renal function worsened on patients with type 2 HRS. Patients with lower hemoglobin levels had poor prognosis and survival compared with patients with higher hemoglobin levels. Logistic regression analysis showed that lower hemoglobin levels and higher MELD and CTP scores were statistically significant for an onset of type 2 HRS. Conclusion. Renal dysfunction is a frequent complication in patients with end-stage chronic liver disease. The role of anemia in aggravating HRS in patients with cirrhosis is explained by hypoxia that can lead to microcirculatory renal ischemia. Other studies are required to determine if anemia is a precipitant factor for HRS or not. Hepatorenal syndrome is a form of functional renal failure arising from liver cirrhosis secondary to diminished renal blood flow . TypicalAnemia causes microcirculatory tissue hypoxia. This phenomenon is called anemic hypoxia. Renal ischemia is the most common cause of HRS. Renal tissue hypoxia or ischemia can trigger the initial tubular damage .Anemia is frequently observed during the clinical course of cirrhosis. There are rare publications addressing the potential or additive effects of anemia on HRS in cirrhotic patients.Our study aimed to investigate the hematologic findings in patients with cirrhosis to determine the effects of anemia on renal functions in type 2 HRS and if anemia was a potential aggravating factor.A prospective study in which all consecutive patients with cirrhosis were enrolled was performed at a tertiary-level hospital  from January 2019 to June 2019. This study was performed according to the Declaration of Helsinki, following the guidelines for good clinical practice, and the ethics committee approved the study protocol.The diagnosis of cirrhosis was based on the clinical and laboratory signs of hepatocellular failure and portal hypertension such as stellar angioma, hepatic encephalopathy, low serum albumin level, prolonged prothrombin time, ascites, jaundice, esophageal or gastric varices, and irregular hepatic echogenicity of the liver in abdominal US.Etiologies of cirrhosis were various including viral hepatitis, autoimmune hepatitis, primary biliary cholangitis, nonalcoholic steatohepatitis, and unknown etiologies.All data regarding patients were obtained from the medical record. Demographic data , routine hemograms, biochemical, and urinary test results were collected. Models of end-stage liver disease (MELD) and Child–Turcotte–Pugh (CTP) scores were calculated.μmol/L) within 48 h or of ≥50% from baseline within 7 days. No improvement of serum creatinine after two days of diuretics withdrawal and volume expansion with albumin (1 g/kg of bodyweight per day up to a maximum of 100 g/day) is required to exclude prerenal acute kidney injury (AKI) due to volume depletion and retain the diagnosis of HRS on patients with AKI.Postrenal dysfunction causes were excluded. The exclusion criteria were determined as shock, current, or recent treatment with nephrotoxic drugs, proteinuria >500 mg/day, infection, urinary red blood cells >50 per high-power field (HPF), abnormal findings on renal ultrasonography, and patients without complete HRS diagnostic criteria. Acute kidney injury is defined as a serum creatinine increase of ≥0.3 mg/dL . Low serum levels of ferritin led to the diagnosis of iron deficiency anemia. Immune hemolytic anemia was diagnosed by the increased serum lactate dehydrogenase level and direct Coombs positivity. Vitamin B12 and folate deficiencies were the diagnoses when the serum levels were lesser than 150 pg/ml and 5 ng/ml, respectively. All patients were followed in-hospital and out-of-hospital for at least six months.μmol/L) within 48 h or of ≥50% from baseline within 7 daysType 2 HRS group: these patients had a serum creatinine increase of ≥0.3 mg/dL  version 21, IBM SPSS Inc., Chicago, IL, USA). Results were expressed as mean ± standard deviation. Univariate comparisons between groups were performed using chi-squared tests for dichotomous variables. For continuous variables, independent samples p=0.129).Fifty patients  with cirrhosis were included in our study. Only 9 patients fulfilled the criteria of type 2 HRS. Of these 50 patients, 27 (54%) were male and 23 (46%) were female. The mean age of our patients was 62.8 ± 11.9 years (range 14–86), and there was no statistically significant difference between the age of the two groups  were categorized in Grade A, 13 (26%) were in Grade B, and 14 (28%) in Grade C, respectively. Of the total of 9 patients with HRS, 3 (33.3%) were in the CTP-B stage, and 6 (66.7%) were in the CTP-C stage . The majEtiologies of liver disease in patients included in this study were hepatitis B in 20 patients (40%), hepatitis C in 11 patients (22%), nonalcoholic steatohepatitis in 5 patients (10%), primary biliary cirrhosis in 1 patient (2%), sarcoidosis in 1 patient (2%), autoimmune hepatitis in 1 patient (2%), primary sclerosing cholangitis in 1 patient (2%), and finally, cryptogenic cirrhosis in 10 patients (20%).p < 0.001). The mean MCV of the whole study group was 85.76 ± 9.8 fl.In this study group, the mean hemoglobin rate was of 10.24 ± 2.58 gr/dl . The meaHypersplenism was the most common cause of anemia among the various causes, representing 50% of cases followed by iron deficiency anemia in 36.1% and immune hemolytic anemia in 13.9% of cases .There were no statistical differences between the groups in terms of age, gender, and platelet count. The demographic features and laboratory data of the study groups are shown in r = −0.333, p = 0.018; r = −0.403, p = 0.04, respectively), higher MELD and CTP scores , and the presence of ascite , which are indicators of the degree of liver failure. As hemoglobin levels decreased, renal function worsened attested by the increased serum creatinine level in the patients with type 2 HRS. There were no significant correlations between serum creatinine levels, bilirubin level hematocrit, and the presence or not of encephalopathy.In patients with type 2 HRS, there was a correlation between serum creatinine and lower hemoglobin and albumin levels   .p < 0.001). Lower hemoglobin levels showed a significant association with mortality in all patients with HRS type 2 (p = 0.038). CTP and MELD scores were higher in nonsurvivors (Six patients (66.66%) died; three died during hospitalization, while 3 died out-of-hospital. The overall survival was 5.2 months. The mortality was higher in patients with HRS compared with those with no HRS , autoimmune hepatitis (2%), primary biliary cholangitis (2%), primary sclerosing cholangitis (2%), sarcoidosis (2%), nonalcoholic steatohepatitis (10%), and unknown etiologies (20%).According to the CTP score, 23 patients were in the CTP-A stage, 13 in the CTP-B stage, and 14 patients were in the CTP-C stage. Anemia was observed in 72% of patients, comparable to literature .Patients with type 2 hepatorenal syndrome had significantly lower hemoglobin levels compared with non-HRS stable cirrhosis patients. As hemoglobin levels decreased, renal function worsened attested by an increased serum creatinine level in the patients with type 2 HRS. Patients with lower hemoglobin levels had poor prognosis and survival compared with patients with higher hemoglobin levels.Despite the fact that HRS is a functional syndrome, it is still associated with a high mortality and poor prognosis . Renal iIn normal conditions, the liver plays a significant role in the regulation of renal function. Hepatic regulation of renal function involves a hepatorenal reflex which is elicited by an increase in intrahepatic pressure and amino acids in portal venous blood. This reflex leads to a marked decrease in renal blood flow and glomerular filtration . Portal Anemia was more frequent with advanced liver disease. Hemoglobin levels dropped when the MELD score increased . The freA strong positive correlation was observed between anemia and the risk of poor outcomes in patients with end-stage renal disease .In cardiology, anemia and iron deficiency are often associated with less good prognosis on patients with heart failure. Correction of the anemia improves functional capacity and decreases hospitalizations, but does not ameliorate mortality in these patients . AnotherStudies have shown a strong correlation between anemia and negative clinical outcome, worse prognosis, and increased complications. The same conclusions may also be accepted for HRS. Anemia is not considered as one of the recognized factors for HRS. There is a difference between variceal bleeding and anemia. While the first causes acute volume depletion, anemia causes hypoxia. Treating anemia might be beneficial in improving renal functions in patients with HRS associated to other medical treatment modalities of HRS .Renal dysfunction is a frequent complication in patients with end-stage chronic liver disease. The role of anemia in aggravating HRS in patients with cirrhosis is explained by hypoxia that can lead to microcirculatory renal ischemia. Other studies are required to determine if anemia is a precipitant factor for HRS or not.


Long noncoding RNA gastric cancer highly expressed transcript 1 (lncRNA GHET1) is often reported to be abnormally expressed in multiple cancers, but the situation is different in different cancers. Therefore, a meta-analysis is necessary to clarify the value of lncRNA GHET1 as a prognostic indicator in cancer.  Relevant research studies on lncRNA GHET1 and cancer were retrieved from three electronic literature databases of Web of Science, PubMed, and OVID. Meanwhile, hazard ratios (HRs) and 95% confidence intervals (CIs) were calculated to explore the relationship between lncRNA GHET1 expression and survival of cancer patients. The odds ratios (ORs) and 95% CIs were calculated to assess the association of lncRNA GHET1 expression with pathological parameters of cancer patients.  The meta-analysis included a total of 11 studies involving 714 cancer patients. The pooled HR suggests that high lncRNA GHET1 expression is associated with poor overall survival. In addition, high expression of lncRNA GHET1 was found to be associated with larger tumor size, poor histological grade, high tumor stage, lymph node metastasis, and distant metastasis.  High lncRNA GHET1 expression can predict poor survival and pathological parameters. And lncRNA GHET1 could serve as a new indicator in multiple cancers. Cancer is still the main disease that threatens people's lives and health. The American Cancer Society estimates new cancer cases and deaths in the USA every year, and 1,806,590 new cancer cases and 606,520 cancer deaths are expected in the USA according to Cancer Statistics in 2020 [Long noncoding RNA (lncRNA) is a type of RNA with a length >200 nt and lacking or no open reading frame (ORF). The number of lncRNA is much lower than that of encoding protein genes (mRNA). More and more studies show that lncRNA has important functions such as transcription and epigenetic regulation in diseases . At the lncRNA gastric carcinoma highly expressed transcript 1 , a recently identified lncRNA discovered in 2014, is located on chromosome 7q36.1 in the human genome . In the First, this systematic review and meta-analysis is reported in accordance with the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) Statement. Then, articles regarding lncRNA GHET1 as a prognostic biomarker for the survival of cancer patients were systematically retrieved in three online databases  from inception to January 20, 2019, by two authors independently in accordance with the standard guidelines for meta-analyses , 14. TexAll the included studies were evaluated, and data were extracted by two authors independently. The study inclusion criteria were as follows: (1) studies in which all tumors were confirmed by histological or pathological examinations; (2) studies in which lncRNA GHET1 expression in human tumor tissues was measured and patients were grouped in accordance with the lncRNA GHET1 expression level; (3) studies that performed statistical analyses on the pathological or patient survival parameters concerned with lncRNA GHET1 expression, such as overall survival (OS), high tumor stage (HTS), lymph node metastasis (LNM), or larger tumor size (LTS); and (4) studies with sufficient original data.Besides, the study exclusion criteria were as follows: nonhuman studies and non-English studies; editorials, reviews, and expert opinions as well as letters; database analysis without original data; and studies concerning the functions of lncRNA GHET1 only but without the molecular mechanism analysis.Relevant data were examined and extracted from the original articles by two reviewers independently. Any disagreement between them was solved through the consensus with a third reviewer. A series of data were collected for this meta-analysis, including surname of the first author, publication year, country, tumor type, sample size, number of patients with LTS, poor histological grade (PHG), HTS, LNM, and distant metastasis (DM), reference gene and detection method of lncRNA GHET1, HRs and 95% CIs of elevated lncRNA GHET1 for OS, threshold of lncRNA GHET1 expression level, and the Newcastle–Ottawa scale (NOS) score.Q and I2 tests were conducted to detect the potential heterogeneity, the results of which indicated significant heterogeneity in this meta-analysis (I2 ≥ 50% and P < 0.1) [The Stata version 12.0 software was adopted for all statistical analyses. In addition, the P < 0.1) . TypicalP < 0.1) . MoreoveDetails of the study selection process are displayed in I2 = 0.0% and PQ = 0.676). Our results indicated that OS was evidently related to lncRNA GHET1 . The pooled OR was 2.80 upon analysis . Afterwards, a sensitivity analysis was carried out among all the included studies, and the heterogeneity had disappeared after excluding the study by Xia et al. (I2 = 4% and PQ = 0.399), with the OR between high versus low lncRNA GHET1 expression groups of 3.28 (95% CI: 2.30–4.67) . The OR between the high and low lncRNA GHET1 expression groups was 2.36  in the absence of obvious heterogeneity. Our results indicated that HTS in cancer patients was associated with high lncRNA GHET1 expression . The OR of the high versus low lncRNA GHET1 expression groups was 3.83 . In addition, the OR of high versus low lncRNA GHET1 expression groups was 3.90 (95% CI: 2.12–7.15; In this meta-analysis, the correlation of HTS with lncRNA GHET1 expression was examined in five eligible studies recruiting 294 patients. The fixed-effects model was selected in this study since there was limited heterogeneity (z| = 0.536; z| = 0.917 and P > |t| = 0.984; z| = 0.548 and P > |t| = 0.608; z| = 0.133 and P > |t| = 0.243; z| = 0.174 and P > |t| = 0.096; z| = 0.462 and P > |t| = 0.285; To evaluate the potential publication bias, Begg's funnel plot and Egger's test were carried out in this meta-analysis. As could be observed from As mentioned in our previous meta-analysis , cancer β-catenin signaling pathway [According to previous studies, lncRNA GHET1 has been proved to be an important oncogene in various human cancers, including NSCLC, HCC, BC, ESCC, HNC, BRC, GC, and PC –26. It c pathway , 34, and pathway .Drug resistance is the main obstacle to successful chemotherapy for patients with cancer. The relationship between lncRNA GHET1 and drug resistance has been studied in gastric cancer and other solid tumors. It is to provide preclinical data for the translational research of lncRNA GHET1 in cancer treatment. Li et al.  found thTo further investigate the underlying mechanism, target, and function of lncRNA GHET1 in cancer, the potential targets, pathways, and related miRNA of lncRNA GHET1 had been systematically reviewed . As mentIn this meta-analysis, data collected from the eleven eligible studies involving seven hundred and fourteen cancer patients were analyzed. Subsequently, a fixed- or a random-effects model was adopted depending on the results of heterogeneity analysis. For cancer patients, high lncRNA GHET1 expression might potentially serve as an indicator of poor prognosis. A significant difference in OS was detected between the high and low lncRNA GHET1 expression groups after pooling HRs from Cox multivariate analyses. In addition, high lncRNA GHET1 expression was found to be dramatically correlated with poor OS in various cancer types. Furthermore, high lncRNA GHET1 expression was also remarkably related to the clinicopathological parameters in cancer patients, including LTS, PHG, HTS, LNM, and DM. To sum up, findings in this study indicated that lncRNA GHET1 might serve as a valuable prognostic biomarker for the poor prognosis of most cancers.When interpreting the conclusions of the current meta-analysis, several limitations should be taken into consideration. Firstly, all studies included in this meta-analysis were retrieved from three online databases; as a result, it was likely that some related papers might be missed. Secondly, after searching the database according to the inclusion and exclusion criteria, only ten studies came from China, so more research from other countries in the future is still needed to prove our conclusions. Thirdly, the cutoff values of the high- and low-expressed lncRNA GHET1 in these studies were not consistent. In addition, as bone spreading is also an important clinical feature related to tumor progression and prognosis, the role of lncRNA GHET1 in tumor-associated bone skeletal diffusion can be studied in the future. Finally, other factors that may affect cancer prognosis, such as treatment and tumor subgroup, were not included in this study. Because these included studies do not include these data, this may be an inherent shortcoming of this systematic review and meta-analysis.To sum up, high lncRNA GHET1 expression in a series of cancers is associated with poor OS, LTS, PHG, HTS, LNM, and DM. As a result, lncRNA GHET1 can be used as a promising biomarker to predict tumor metastasis and prognosis in cancer patients.


Somatic, physiological, and biochemical variables were measured in four training phases differing in exercise load profile: transition, general, specific, and competition. Significantly highest values of RBC adenylate energy charge , ATP-to-ADP and ADP-to-AMP ratios (p ≤ 0.05), and plasma levels of adenosine  were noted in the competition phase in the EN and SP, but not in the RE group. Significantly lowest plasma levels of adenosine diphosphate , adenosine monophosphate , inosine , and hypoxanthine  accompanied by higher erythrocyte hypoxanthine-guanine phosphoribosyltransferase (HGPRT) activity (p ≤ 0.001), were observed in the competition phase in both athletic groups. No significant alterations were found in the erythrocyte concentration of guanine nucleotides in any group. In conclusion, periodized training of competitive athletes’ results in a favorable adaptation of RBC metabolism. The observed changes cover improved RBC energy status (increased AEC and ATP/ADP ratio) and reduced purine loss with more efficient erythrocyte purine pool recovery . This study aimed to assess the changes in red blood cell (RBC) energy status and plasma purine metabolites concentration over a one-year training cycle in endurance-trained (EN;  Mammalian erythrocyte (RBC), a unique cell without a nucleus, serves as a model cell in many studies. Apart from its main functions, such as carrying oxygen and carbon dioxide, and controlling the acid-base balance level, erythrocytes also exert other roles. They take part in nitric oxide (NO) metabolism, influence blood rheological properties, and exert the erythrocrine function by releasing vast amounts of ATP and other bioactive molecules ,2. All tThe RBCs’ vitality, resilience, and functioning rely on their energy metabolism, mainly glycolysis, which is the only source of adenosine triphosphate (ATP) in RBC . Since AThere are scant studies reporting changes in purine nucleotide catabolite concentration in human and animal erythrocytes in response to single bouts of exercise ,9,10,11.The research conducted so far demonstrated that there were significant differences between trained and sedentary individuals in erythrocyte energetics  , which mThus far, changes in erythrocyte energy status and purine nucleotide catabolites concentration were not yet explored also during a longer period or under prolonged training regime. Reports based on one-year training cycles only focused on changes in plasma hypoxanthine (Hx) concentration and erythrocyte hypoxanthine-guanine phosphoribosyltransferase (HGPRT) activity ,14,15. IThe research on the link between erythrocyte energetics and whole-body functioning, e.g., during various physical efforts, may give new insight into some aspects of exercise and training physiology. We expect that the indicators of RBC energy status and the level of ATP metabolites are sensitive to changes in training specificity and exercise loads and, thus, they could be used as a valuable tool indicating the training status in athletes. Thus, this study aimed to assess whether the change in training loads during a periodized one-year cycle is associated with the changes in erythrocyte energy status and plasma purine metabolites concentration in highly-trained athletes. We hypothesized that the erythrocyte energy status will adapt to the alterations in training loads throughout the phases of the annual training cycle. p ≤ 0.001) between consecutive phases of the annual training cycle were observed in the EN group for weight, body mass index (BMI), and maximal oxygen uptake (VO2max), while in the SP group for weight, BMI, VO2max, Hct, and lactate (LA) at rest.The somatic and exercise characteristics of studied participants in consecutive phases of the annual training cycle are shown in p ≤ 0.05). In all study examinations, there were significant differences between analyzed groups in weight, BMI, Hb concentration, Hct, and VO2max . LA concentration at rest was significantly different between groups in the general and competition phases (p ≤ 0.05).During the studied training cycle, only weight and Hb changed significantly in the RE group (p ≤ 0.05). These changes occurred mainly between transition and specific and between transition and competition phases, as measured at rest, immediately post-exercise, and after 30 min of recovery. The differences between measurements at rest, post-exercise and recovery were observed in endurance athletes (ADP and AMP), sprinters ) and in the recreationally-trained group (only AMP) (p ≤ 0.05). p ≤ 0.001), TAN (p ≤ 0.05), and IMP (p ≤ 0.001) in all examinations, and in AMP concentration (p ≤ 0.05) in the specific and competition phases. The differences between EN and RE groups were significant for ATP in the transition and specific phases (p ≤ 0.001), for ADP and AMP in the specific and competition phases , and for IMP concentration (p ≤ 0.001) in the general, specific, and competition phases of the annual training cycle. Endurance athletes differed significantly from sprinters only in ATP concentration in the transition, specific, and competition phases of the annual training cycle. There were significant differences between sprinters and recreational runners in the level of ATP . Significant differences were also noted in the RE group . As indicated in p-values ranged from <0.001 to 0.004): (i) EN vs. RE group , and (ii) SP vs. RE group . There were no significant differences between the EN and SP groups. Following significant between-group differences occurred in the level of AEC and ATP/ADP ratio (p ≤ 0.001). In the SP group, the differences were observed between the transition and general and between the general and specific training phases (p ≤ 0.001). There were no differences in the level of erythrocyte HGPRT activity in the annual training cycle in the RE group. There were significant differences in erythrocyte HGPRT activity between all analyzed groups in each training phase (p ≤ 0.001). The between-group differences in resting erythrocyte HGPRT activity in consecutive training phases are shown in p ≤ 0.001) and Hx (p ≤ 0.001) concentrations measured immediately after exercise and after 30 min of recovery. There were also significant changes in Ado concentration between the transition and competition phases in the EN group in all three measurement points and in the SP group immediately after exercise and 30 min of recovery. Significant changes between rest and post-exercise values as well as between post-exercise and recovery values were observed in all measurement points (p ≤ 0.001) in the EN and SP groups. In the RE group, significant changes were shown for Ino, Hx , and Guo . The concentration of Ado changed significantly only between post-exercise and 30-min recovery values in the SP  and in the RE group . p ≤ 0.001). Significant differences (p ≤ 0.002) occurred between SP and RE groups in the level of Ado in all training phases and between EN athletes and RE group in Ino concentration in the competition phase of the annual training cycle (p ≤ 0.002). There were significant differences in plasma Hx concentration between all analyzed groups in each training phase  in highly-trained athletes, indicate that the adaptation to high-intensity training loads plays a key role. Such an adaptation signifies a more effective ATP production and probably an increased activity of anabolic reactions dependent on ATP inflow, including repair mechanisms. Thus, enhancing the RBC energy potential may be considered as one of the links in the process of adaptation to increased tolerance to high-intensity exercise.Results obtained in our study suggest that the longitudinal changes observed in athletes are mainly related to an increase in the energy potential of the RBC and the enzymatic activity in the purine resynthesis pathway. The differences in erythrocyte energy status between athletes and controls are most likely due to training-induced increments in RBC turnover, resulting in shorter RBC lifespan and a higher proportion of younger RBCs in trained individuals ,3,5,6. Guaninetriphosphate (GTP) plays a key role in protein synthesis, it is a substrate in the cyclic guaninemonophosphate (GMP) synthesis and a potential source of energy. However, due to the significantly lower concentration in the erythrocyte, guanine purines exert a smaller impact on erythrocyte bioenergetics compared to adenine ,18.In our study, in line with previous ones ,10, the salvage pathway. Other researchers observed similar responses in the skeletal muscle after a strenuous short-term exercise [We found that in the control group the activity of erythrocyte HGPRT, an enzyme involved in RBC purine resynthesis, did not change throughout the investigated training period and that it remained significantly lower compared to the athletes. A similar difference was previously shown between sedentary people and trained rowers . In our exercise ,20. It iIn highly-trained athletes, but not in the control group, the increased amount of intense training loads in the subsequent phases of the annual training cycle resulted in adaptive changes in plasma purine derivatives concentration. The significantly lowest levels of plasma Ino and Hx in the competition phase indicate either their reduced release from the skeletal muscle ,21 or thAn important phenomenon we observed was the significantly higher Ado concentration in endurance and sprint-trained athletes in the competition compared to the transition phase. Because such changes were not found in the recreationally-trained group, it seems that the inclusion of high-intensity workouts, including frequent hypoxia episodes, leads to an increase in Ado production, which source in hypoxia is the extracellular AMP hydrolysis catalyzed by ecto-5’-nucleotidase . This asThe strength of this study is the inclusion of two homogenous groups of top-level athletes from national teams, trained by the same coaches. Training loads were precisely scheduled and their volume and intensity changed according to the aim of each training phase. To our best knowledge, this is the first study to comprehensively analyze purine metabolism (13 purine nucleotides) in a longer period in highly-trained athletes of two distinct specializations.There are also some limitations. Elite athletes of different specializations have different genetic predispositions and are subject to sports selection at an early age. In future research, it would be interesting to analyze to what extent the observed changes and differences in RBC energy status and purine metabolism relate to genetic factors. It would be also beneficial to rate the activity of glycolytic enzymes, which is the only metabolic pathway that produces ATP in the human red blood cell. The conversion of intracellular AMP and ADP into ATP requires two reactions, i.e., the phosphorylation of AMP to ADP by adenylate kinase and the phosphorylation of ADP to ATP by pyruvate kinase and phosphoglycerate kinase. Hence, it is possible that long-term training can lead to specific adaptive changes in the activity of enzymes in the ATP-generating pathway. The combined assessment of enzyme activity, especially regulatory enzymes, the blood concentration of purine nucleotides, and their metabolites would allow for a better assessment of RBC energy metabolism and a deeper insight into the mechanisms behind the observed differences. The limiting aspect of this study may also be the lack of precise control of dietary intake in the recreationally-trained group. We have familiarized them with standard nutritional recommendations for physically active people, however, we did not precisely control to what extent they adhered to them.n = 11) included triathletes, aged 22.9 ± 2.12 (20–26) years, practicing competitive sport for 8.27 ± 1.62 (6–11) years, specializing in the standard Olympic distance . The second athletic group included sprint-trained men , aged 23.55 ± 3.27 (20–30) years, practicing competitive sport for 8.18 ± 0.98 (7–10) years, specializing in the 100 m, 200 m, and 4 × 100 m relay races. The control group consisted of recreationally-trained runners , aged 22.73 ± 2.15 (20–26) years, competing in half marathons, marathons, or road/cross-country running, but not subjected to a periodized training regimen. Over the research period, there were no significant differences between the studied groups as regards age (p = 0.737), body height , and training experience in competitive athletes (p = 0.875). Both EN and SP athletes adhered to unified nutrition recommendations made by the dietician of the national team, depending on the training phase of the one-year cycle. The control group followed standard nutritional recommendations for physically active people [The study included three groups of male adult athletes, two of which comprised endurance- and sprint-trained athletes from the Polish national teams, who competed at the national and international level. The endurance-trained group  and conducted following the Helsinki Declaration.The study procedure was adjusted to the annual training plan of EN and SP athletes. All measurements, in both athletes and recreationally-trained participants, were performed in four characteristic training phases as described by Bompa and Buzzichelli  (each ti2max) was measured in the morning using an incremental treadmill test . Initially, the exercise protocol included standing still for three min and walking for three min at a constant speed of 4 km/h. After this introductory part, the speed was increased to 8 km/h and then every 3 min by 2 km/h until the volitional exhaustion was reached. Oxygen consumption was measured with the use of a portable breath-by-breath ergospirometer . VO2max was considered achieved if at least three of the following criteria were met: (i) a plateau in VO2 despite an increase in running speed, (ii) blood lactate concentration ≥9 mmol/L, (iii) respiratory exchange ratio ≥1.10, and (iv) heart rate ≥ 95% of the age-predicted HRmax [A SECA 285 digital stadiometer  was used to measure height and weight. Maximal oxygen uptake : (i) before exercise, (ii) immediately after exercise (at exhaustion), and (iii) after 30 min of recovery. Blood samples were taken into two separate tubes with EDTA (2.7 mL) and lithium heparinate (4.9 mL) as anticoagulants . The first tube was used for the determination of hematocrit (Hct) value and hemoglobin (Hb) concentration. The second tube was used to assess all other compounds: lactate (LA) in whole blood, erythrocyte purine nucleotides , GMP), erythrocyte hypoxanthine-guanine phosphoribosyltransferase (HGPRT) activity, plasma adenosine (Ado), inosine (Ino), guanosine (Guo), and hypoxanthine (Hx).® 18 . LA concentration was assayed immediately, before the separation of the erythrocytes from plasma, in whole blood (20 μL) using the spectrophotometric enzymatic method . Hct and Hb measurements were carried out in 10 µL of aspirated blood using the 18-parametric automated hematology analyzer Mythicg, 5 min, 4 °C). Plasma and buffy coat were removed. Then, plasma was divided into aliquots and immediately deep-frozen at −80 °C until the analysis. Erythrocytes were washed three times with buffered 0.9% NaCl solution and centrifuged each time . After the final wash, the resulting erythrocyte pellet was resuspended with a small volume of PBS. The isolated and washed erythrocytes were collected in Modulohm glass capillaries  to obtain hematocrit values (Hct). Hct values were determined in duplicate by the standard microhematocrit method. Next, the samples of washed erythrocytes were deproteinized with an equal volume of 1.3 mol/L HClO4, mixed, and then centrifuged at 16,000× g for 5 min at 4 °C. The supernatant (600 μL) was neutralized with 130–160 μL of 3 mol/L K3PO4 (to pH 5–7). The samples were centrifuged again under the same conditions as previously, and the supernatant was stored at −80 °C until the analysis.To isolate erythrocytes, the whole blood in the second tube was centrifuged  within 3 min after drawing a sample (1000× All subsequent measurements in RBC and plasma were performed using high-performance liquid chromatography (HPLC) with UV–VIS detection  according to the method developed by our group, as described in detail by Smolenski  and usedTo determine ATP, ADP, AMP, IMP, GTP, GDP, and GMP, 100 μL of supernatant was injected into the sample loop. Purine nucleotides were separated using a gradient elution system at a flow rate of 1 mL/min. Peaks were detected by absorbance at 254 nm. After conversion to Hct, the intra-erythrocyte concentrations of purine nucleotides were expressed as μmol/L RBC. The values of total adenine nucleotide pool (TAN = ATP + ADP + AMP), total guanine nucleotide pool (TGN = GTP + GDP + GMP) were calculated. Adenylate and guanylate energy charge ratios  were calculated using the following formulas:2, and 5 mmol/L Hx was prepared. After 5 min of pre-incubation at 37 °C, 25 μL of the previously prepared lysate was added. The samples were incubated for 15 min at 37 °C. Reactions were stopped by adding 0.8 mol/L of HClO4 and immediately placed on ice. HGPRT activity was calculated from the IMP concentration increase during 15-min incubation and expressed as nmol IMP/mg Hb/h. Drabkin’s method was used for the determination of hemoglobin concentrations in the erythrocyte lysates.HGPRT activity was determined in the RBCs lysates. After Hct measurement (standard microhematocrit method), the previously washed erythrocytes were diluted with the Tris buffer  to achieve the Hct value equal to 20%. Erythrocytes prepared in this manner were frozen and thawed twice to increase the lysis of cells. Hemolysates were stored at −80 °C until analysis. To determine the HGPRT activity, 250 μL of the reaction mixture containing 2.5 mmol/L PRPP in 125 mM Tris buffer (pH 7.4), 13 mmol/L MgCl4) and once more centrifuged . An acid supernatant was neutralized with 3 mol/L potassium phosphate K3PO4, centrifuged , and stored at −80 °C before analysis. One hundred microliters of supernatant was injected into the sample loop. The concentrations of Guo, Ino, Ado, and Hyp were expressed as μmol/L of plasma.The obtained plasma was deproteinized with 1.3 mol/L perchloric acid , adenine nucleotides (η2 = 0.18–0.97), guanine and pyridine nucleotides (η2 = 0.20–0.74), and descriptive and exercise characteristics (η2 = 0.24–0.99). The statistical power of ANOVA analyses was 0.63–1.0 for purine nucleotides, 0.58–1.00 for adenine nucleotides, 0.64–0.99 for guanine and pyridine nucleotides and 0.67–1.00 for descriptive and exercise characteristics. All statistical analyses were performed using Statistica 13.3 software . The sample size was a priori estimated based on the assumption that effect size will be at least medium. Using an α-level of 0.05, a power (1-β) of 0.80, it was calculated that at least eight participants in a single group would be needed to detect a significant change or differences in purine metabolite concentration (RBC and plasma) and erythrocyte HGPRT activity . All values are presented as mean ± standard deviation (SD). The Shapiro–Wilk test confirmed the normal distribution of analyzed variables, therefore further analyses were performed using parametric tests. The results obtained during the consecutive phases of the annual training cycle were used to estimate longitudinal changes in the concentration of purine and pyridine nucleotides in red blood cells. For this purpose, the analysis of variance was used. To track the changes over time, the multi-factor analysis of variance (ANOVA/MANOVA) for repeated measures was used. A one-way analysis of variance (ANOVA) was used to determine differences between studied groups. The post hoc Scheffe test was used to determine the significance of the differences within and between consecutive study examinations and between three analyzed groups. The level of significance was set at In conclusion, we demonstrated that in competitive athletes an increment in RBC energy status occurred across the annual training period, as assessed by the increments in AEC and ATP/ADP and ADP/AMP ratios. This suggests that the red blood cell metabolism adjusts to the increased physical requirements of the consecutive training phases. Additionally, the increased enzymatic activity (HGPRT) of the purine resynthesis pathway and the accompanying gradual reduction in plasma purine metabolites  concentration indicate increased capacity to reduce purine loss and recover erythrocyte purine pool. The concurrent training phase-dependent increment in plasma Ado concentration in athletes supports this mechanism.


Until now, only two prospective cohort studies have investigated dietary fiber intake in relation to risk of chronic obstructive pulmonary disease (COPD), but neither examined long-term fiber intake. Both studies reported that total fiber intake was associated with decreased COPD risk; however, results for specific fiber sources were inconsistent. Thus, we prospectively evaluated the association between baseline and long-term intake of dietary fiber and COPD risk in a population-based prospective cohort of 35,339 Swedish women.Dietary fiber intake was assessed in 1987 and 1997 with a food frequency questionnaire. Cox proportional hazard regression models were used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs).≥ 26.5 g/day) had a 33-fold (95% CI 23.6–46.6) and tenfold (95% CI 7.0–16.3), respectively, higher risk of COPD.During follow-up (2002–2014), 1557 COPD cases were identified via linkage to the Swedish National Patient Register. Long-term high dietary fiber intake (≥ 26.5 vs. < 17.6 g/day) was associated with a 30% (95% CI 17–41%) lower risk of COPD. For specific fiber sources, cereal  and fruit fiber , but not vegetable fiber intake  were associated with lower COPD risk. Current and ex-smokers with low long-term total fiber intake (< 17.6 g/day) compared to never smokers with high intake (Our findings indicate that high fiber intake is a modifiable lifestyle factor which may decrease COPD risk primarily in current and ex-smokers.The online version of this article (10.1007/s00394-019-02038-w) contains supplementary material, which is available to authorized users. Globally, chronic obstructive pulmonary disease (COPD) is predicted by the World Health Organisation to become the third most common disease-related cause of mortality by the year 2030 . ExposurUntil now, only two prospective cohort studies have investigated dietary fiber intake in relation to risk of COPD development. The first was conducted in two cohorts of US women and men  and the In the current study, we hypothesized that intake of dietary fiber is a beneficial lifestyle choice that reduces the risk of COPD. To address this hypothesis, we investigated the associations between long-term total and specific fiber intake and incidence of COPD in the population-based prospective Swedish Mammography Cohort.n = 90,303) were invited to a mammography screening and completed a food-frequency questionnaire. Among the 66,651 (response rate = 74%) women who returned the questionnaire, we excluded those with missing or incorrect national identification number (n = 1988), history of cancer , and improbable total energy  . The completed questionnaires were received from 39,227 women (response rate = 70%). From these women, we excluded those with missing or incorrect national identification number (n = 243), who were diagnosed with cancer (n = 1717) or COPD before 1998 , and those with improbable total energy intake (n = 489). Additionally, due to a potential under-diagnosis in the first years of follow-up (1998–2001) and the necessity of a 4-year lag period introduction , women with COPD diagnosis between 1998 and 2001 (n = 160) or those who died during these 4 years (n = 977) were excluded as well. Thus, the final analytic cohort consisted of 35,339 women, aged 61.5 ± 9.1 years, who were comparable to the general Swedish female population in relation to age distribution, education, and BMI distribution [The Swedish Mammography Cohort (SMC) was established between 1987 and 1990 when all women living in Uppsala and Västmanland counties in central Sweden, and born during 1914–1948 (ncer n = 17 or COPData about food consumption were collected by food frequency questionnaires (FFQs) based on 67 food items in 1987 and 96 food items in 1997. Participants were asked about their average food consumption during the past year based on eight predefined frequency categories, ranging from “never/seldom” to “4 times per day” in 1987, and from “0 times per month” to “3 + times per day” in 1997. Dietary fiber intake was estimated by multiplying the frequency of intake of each food item by the fiber content   using agIn the present study, information on covariates collected on the 1997 questionnaire was used as baseline. Women were asked about education level, weight, height, time per day spent on walking and/or cycling, smoking status, and alcohol consumption. Body Mass Index (BMI) was calculated by dividing the weight in kilograms by the square of the height in meters. Participants reported average number of cigarettes smoked per day at different ages , and indicated when they started and stopped smoking (only if this event occurred). Pack-years of smoking were calculated by multiplying the number of cigarettes smoked per day by number of years smoked, respectively, for each age category.Linkage to the Swedish National Patient Registers (inpatient and outpatient) and the Swedish Cause of Death Register was performed using the unique personal identification number assigned to each Swedish resident. COPD diagnoses were identified in accordance with the International Classification of Diseases and Related Health Problems, 10th Revision (ICD-10 code: J44). Incident COPD cases were defined as the first diagnosis of COPD in the Swedish Patient Register (listed either as the primary or any diagnosis position) or in the Cause of Death Register (only the primary position). Taking into account that both medical registers in Sweden and several guidelines for the diagnosis and management of the COPD were created in the early 90 s, there was a potential of under-diagnosis in the cohort , 24. Fur2), walking/cycling , smoking status and pack-years of smoking , energy intake , and alcohol intake . For specific fiber sources, intakes of fiber from cereals, fruits, and vegetables were mutually adjusted by inclusion in the same multivariable model. Missing data on smoking status (1.8%), educational level (0.3%), BMI (1.7%), and walking/cycling (7.8%) were included as separate categories. Moreover, we investigated the combined effect of long-term total dietary fiber intake with smoking status  where never-smoking women with the highest fiber intake were considered as the reference group.Women were followed from 1 January 2002 to 31 December 2014, and until COPD diagnosis, death, or the end of follow-up, whichever occurred first. Hazard ratios (HRs) and 95% confidence intervals (CIs) of COPD incidence were estimated using Cox proportional hazard regression models. Participants were assigned to quintiles of baseline energy-adjusted total dietary fiber intake , and the same cut-points were used for long-term fiber intake. For specific fiber intake (both baseline and long term), participants were assigned to quintiles of energy-adjusted consumption. For all dietary fiber exposures, the lowest quintile served as the reference group. Multivariable models were adjusted for: age , education , BMI  on COPD risk. The shape of the association between risk of COPD and long-term total dietary fiber intake was investigated using restricted cubic-splines with three knots  . This meIn 1997, the energy-adjusted mean total dietary fiber intake was 22.2 ± 5.5 (median 21.8) g/day and the contribution of cereal, fruit and vegetables to total fiber intake was 59.1%, 23.2%, 17.5%, respectively. The Spearman correlation coefficients between total fiber intake and cereal, fruit, and vegetable fiber were 0.57, 0.53, and 0.51, respectively. Compared to women in the lowest quintile of total dietary fiber intake (< 17.6 g/day), those in the highest quintile (≥ 26.4 g/day) were less likely to be ever smokers and more likely to walk and/or cycle more than 20 min/day , 1557 incident cases of COPD were identified. An inverse association between total baseline dietary fiber intake and risk of COPD development was observed . The HRs in years 2002–2008 and 2009–2014 were consistent; between extreme quintiles of total fiber intake (≥ 26.5 vs. < 17.6 g/day) were 0.75 (95% CI 0.60–0.94) in 2002–2008 and 0.81 (95% CI 0.65–1.00) in 2009–2014. In 1998–2001, a statistically significant association between dietary fiber intake and COPD risk was not observed . Taking into account the lower number of identified COPD cases in 1998–2001, we hypothesized that the lack of association was at least partly the result of under-diagnosis of COPD incidence during that period.P value for non-linearity = 0.005  and 10 years before baseline (1987), women in the highest vs. lowest quintiles of fiber intake had a 30% (95% CI 17–41%) lower risk of COPD  comparing women in the highest vs. the lowest total dietary fiber intake.For specific fiber sources, we observed an inverse association between risk of COPD and long-term cereal and fruit fiber intake Table . Women wP interaction = 0.79).As smoking is the main risk factor for developing COPD, we conducted stratified analysis by smoking status Table . CurrentTo investigate the combined effect of long-term dietary fiber intake and smoking status on risk of COPD, a comprehensive analysis which included current, ex-smokers and never smokers in one model was performed Fig. . In compIn this prospective cohort study of women, we found that high dietary fiber intake was inversely associated with the risk of COPD, and the risk was decreased in all categories of smoking status—current and ex-smokers, as well as never smokers. For specific fiber sources, cereal and fruit fiber, but not vegetable fiber, was inversely associated with COPD incidence.P trend = 0.03) [P trend < 0.0001) [1), forced vital capacity (FVC) and FEV1/FVC ratio.Our findings confirm and expand upon the results from two previous prospective cohort studies  which i 0.0001)  lower ri 0.0001) , where h1, FVC and FEV1/FVC ratio, while those with the highest vs. the lowest “Prudent” dietary pattern had higher values of spirometry parameters [Moreover, some previous studies have examined the associations between dietary patterns and risk of COPD. A “Prudent” dietary pattern  which is characterized by high amounts of dietary fiber, was inversely associated with COPD risk –29. Morerameters . ResultsConsidering sources of dietary fiber intake, our results confirmed findings of previous studies that higher consumption of cereal fiber was associated with lower risk of COPD , 16. AmoIn contrast to a previous study of Swedish men , we did Various potential mechanisms involving a role of dietary fiber in the modulation of systemic tissue inflammation may explain the inverse association between dietary fiber intake and risk of COPD. Prior studies have demonstrated that higher dietary fiber intake decreased the concentration of pro-inflammatory mediators such as IL-6 and CRP –11. ResuIt seems feasible that dietary fiber may have a beneficial impact on reducing oxidative stress in the lungs via the antioxidant capacity of fruit and vegetables, thus, lowering the risk of COPD. Antioxidants such as polyphenols and carotenoids traverse the small intestine in conjunction with dietary fiber and may release the fiber matrix exclusively in the colon through the action of the microbiota . LinkageIn light of the considerations above, dietary fiber seems to be a unique and important nutrient in the prevention of COPD, especially among smokers. Due to the modulation of the innate immune system via the gut–liver–lung axis and reduction in oxidative stress via enhancing bioaccessibility of antioxidants from fruits and vegetables, dietary fiber appears to be unique, even when compared to other nutrients with anti-inflammatory and anti-oxidative properties.As strengths of this study, the following aspects should be considered: a large population-based prospective cohort and a large number of COPD cases which allowed sufficient power for comprehensive statistical analyses. This is the first study which examined long-term dietary fiber intake in relation to COPD incidence. Data on fiber intake had high validity . MoreoveOur study strengthens and expands previous evidence that, in addition to smoking cessation, intake of dietary fiber as a modifiable lifestyle factor may reduce the risk of COPD for both ex- and current long-term smokers. Therefore, our results support the recommendation to increase consumption of products high in dietary fiber especially among smokers. Furthermore, these findings indicate the need to conduct intervention studies on the clinical utility of recommending intake of at least 25 g of fiber/day to reduce the risk of COPD among long-term smokers.Supplementary material 1 (PDF 156 kb)Below is the link to the electronic supplementary material.


An increasing number of techniques have been used for prenatal diagnosis of genetic abnormalities. Our initial objective was to explore the value of the BACs-on-Beads (BoBs) assay for the prenatal diagnosis of aneuploidies and microdeletion/microduplication syndromes in Quanzhou, Southeast China.A total of 1409 pregnant women with high-risk factors for chromosomal abnormalities admitted to Quanzhou Women’s and Children’s Hospital were enrolled in this study. BoBs assays and karyotype analyses were conducted for all subjects. Subsequently, chromosome microarray analysis (CMA) or fluorescence in situ hybridization (FISH) was performed to validate the findings.In this study, karyotype analysis and BoBs assay failed in 4 cases, and 2 cases, respectively. A total of 1403 cases were successfully analyzed, with success rates of 99.72% (1405/1409) and 99.85% (1407/1409) for karyotype analysis and Bobs assay, respectively. BoBs assay rapidly detected chromosomal aneuploidies in line with the karyotyping data. Additionally, 23 cases of microdeletions/microduplications were detected by BoBs assay but missed by karyotyping, including 22q11.2 microdeletions/microduplications, 5p15.32p15.33 microdeletion, Xp22.31 microdeletions/microduplications, Xq27.3 microdeletion, and Yp11.2 and Yq11.22q11.222 microduplication. In comparison with karyotyping, fewer mosaicisms were identified by BoBs assay. A high detection rate of chromosomal abnormalities was observed in the high-risk group during noninvasive prenatal testing (NIPT) (41.72%) and the abnormal ultrasound group (13.43%).BoBs assay can be used for the rapid and efficient prenatal diagnosis of common aneuploidies and microdeletion/microduplication syndromes. Moreover, the combined use of BoBs assay and karyotyping in prenatal diagnosis may allow for a more effective detection of chromosomal abnormalities.The online version contains supplementary material available at 10.1186/s12884-021-03589-9. The preo 106 bp . To dateo 106 bp . Patiento 106 bp . Furthero 106 bp .In this study, a total of 1409 pregnant women with high risk factors were enrolled for prenatal diagnosis, BoBs assay was performed to explore its application value in terms of detecting aneuploidies and microdeletion/microduplication syndromes in comparison with karyotyping in Quanzhou, China. This study also provides available data regarding the importance of prenatal diagnosis in the prevention and control of birth defects, and is of great significance in further supporting the application value of BoBs assay in prenatal diagnosis.n = 190), a high-risk NIPT results group (n = 163), an abnormal ultrasound group (n = 134), a high-risk serological screening results group (n = 480), an adverse pregnancy history group (n = 159) and a group of patients with two or more types of high-risk factors (n = 283). All of the participants signed an informed consent. Approval was obtained from the institutional ethics committee of Quanzhou Women’s and Children’s Hospital (2020No.12 and 2020No.31).A total of 1409 pregnant women with high-risk factors for chromosomal abnormalities admitted at Quanzhou Women’s and Children’s Hospital from January 2016 to December 2019 were enrolled in this study. The indications for prenatal genetic evaluation were advanced maternal age (> 35 years), high-risk NIPT results, abnormal ultrasound, high-risk serological screening results, and adverse pregnancy history. The subjects were divided into six groups, including an advanced maternal age group . Additionally, G-band karyotype analysis was performed after staining with Giemsa. Subsequently, 30 metaphase cells were observed for each case, and 5 karyotypes were analyzed. C-banding was conducted when necessary.www.qiagen.com).The subjects’ DNA was extracted from uncultured amniotic fluid cells for BoBs assay and further SNP array verification. DNA extraction of amniotic fluid was performed using a QIAamp DNA blood Kit , following the kit’s handbook , and 9 common microdeletion syndromes according to the manufacturer’s instructions  (Table http://dgv.tcag.ca/dgv), OMIM , DECIPHER (https://decipher.sanger.ac.uk/), and PubMed (https://www.ncbi.nlm.nih.gov/pubmed/), along with other databases.A SNP chromosome microarray (AffymetrixCytoScan™ 750 K) was performed to verify samples with microdeletions/microduplications detected by BoBs assay. A total of 250 ng of genomic DNA was digested with the NspI enzyme, the ends were filled with ligase, and genomic DNA was amplified by polymerase chain reaction (PCR). After the product was purified and prepared for quantitative detection, the fragmentation reaction was carried out to generate fragments of approximately 25-125 bp, which were then labelled with biotin and hybridized with the probe on the chip. The hybridized chip was washed and stained, then analyzed after the fluorescent signal was scanned. The Chromosome Analysis Suite (ChAS) v4.0 software was used to analyze the fluorescence signal in order to determine the sample’s genome-wide variation of the sample. The pathogenicity of copy number variations was interpreted with reference to DGV  according to the manufacturer’s protocol. One hundred amniotic cells were counted and analyzed by fluorescence microscopy  to determine the occurrence of chromosome aberrations.p < 0.05.Statistical analysis was conducted with the SPSS18.0 software . The differences among groups were analyzed using the chi-square test. Statistical significance was determined at A total of 1409 prenatal samples were analyzed, with the mean maternal age being 26.2 ± 3.3 years old and the mean gestational age being 18.3 ± 2.4 weeks. Chromosomal abnormalities were detected by karyotyping and BoBs assay, with 4 cases missed by karyotyping, 2 cases missed by BoBs assay, and 1403 cases successfully analyzed. As illustrated in Table In this study, the success rates were 99.72% (1405/1409) for the karyotyping and 99.85% (1407/1409) for BoBs assay. Moreover, 140 cases of chromosomal abnormalities were identified by either karyotyping or BoBs assay, with a detection rate of 9.98% 140/1403) Moreover, 23 cases of microdeletions/microduplications were detected by BoBs assay Table , includip < 0.001). As delineated in Table Significant differences in detection rates of chromosomal abnormality were observed across groups with different high-risk factors (Chromosomal abnormalities are among the primary reasons for birth defects, with trisomy 21, trisomy 18, trisomy 13 and sex chromosome aneuploidies being the most prevalence entities . KaryotyIn this study, 140 cases of chromosomal abnormalities were identified by the two detection technologies, with a detection rate of 9.98%, which was similar to previous studies , 11. TheNDUFS6 gene, which is associated with mitochondrial complex I deficiency disease [In this study, 23 cases of chromosome microdeletions/microduplications were detected by BoBs assay, including 2 cases of 22q11.2 microduplication syndrome with a 3.1-Mb duplication. Previous studies have reported that 22q11.2 duplications can lead to a clinical phenotype in some subjects, while others exhibit a normal phenotype, indicating incomplete penetrance of the duplication . In some disease . Further disease , but sim disease , 16, 17. disease , 19. In Significant differences in chromosomal abnormality detection rates were observed among the groups with different high-risk factors. Of these groups, the high-risk NIPT results group and ultrasound abnormality group showed high detection rates of chromosomal abnormalities, consistent with other studies –22. HencThis study first investigated the application value of BoBs assay for prenatal diagnosis in Quanzhou, which provides baseline data regarding the combined application of BoBs assay and karyotyping for prenatal diagnosis of chromosomal abnormalities. In this study, we further proved that BoBs assay is a rapid and reliable detection technology for the prenatal diagnosis of common aneuploidies and microdeletion/microduplication syndromes, while its ability to detect chromosomal mosaicisms is limited. Furthermore, the combined use of BoBs assay and karyotyping in prenatal diagnosis may allow for a more effective detection of chromosomal abnormalities.Additional file 1.


The chloroplast genome is a circular molecule of 139404 bp in length, consisting of a 82938 bp large single copy region (LSC), a 12876 bp small single copy region (SSC), and a pair of inverted repeats region (IRa and IRb: 21795 bp each). The GC content of chloroplast genome is 38.9%. The cp genome contains a total of 133 genes, including 86 protein-coding genes, 8 rRNA genes, and 39 tRNA genes. Moreover, phylogenomic analysis showed that D. hamiltonii and D. brandisii clustered together in one branch.Dendrocalamus  Dendrocalamus hamiltonii is a large bamboo species with sympodial rhizome that is classified as genus Dendrocalamus within the family Gramineae and subfamily Bambusoideae, and is widely cultivated in Xishuangbanna, Puer and Lincang, Yunnan province, in China. It is known as the three famous sweet bamboo shoots in the world with D. brandisii and D. asper. The young shoots are sweet and delicious, which are used as vegetables. It is also high-quality building material used by national minority to build houses . Paired-end reads were sequenced by using Illumina Hiseq platform in Sangon Biotech in Shanghai, China, which gained 4.4 Gb of 150 bp paired-end raw reads. The cp genome of D. hamiltonii was assembled using the program GetOrganelle . To determine the phylogenetic relationship of D. hamiltonii, based on 29 cp genomes within the subfamily Bambusoideae, and were downloaded from NCBI. All cp genomes were aligned using the program MAFFT v7.450 , China. The specimen was deposited at Institute of Bamboo and Rattan, Southwest Forestry University (specimen number:SWFU1992015). Total DNA of D. brandisii (GenBank Accession:MN933913) is a circular molecule of 139404 bp in length with GC average content is 38.9%, and consisting of a 82938 bp large single copy region (LSC), a 12876 bp small single copy region (SSC), and two inverted repeats region of 21795 bp. The cp genome of D. hamiltonii contains a total of 133 genes, including 86 protein-coding genes, 8 rRNA genes, and 39 tRNA genes, and the introns were found in 13 genes, six protein-coding genes  and six tRNA genes  have one intron each, one gene (ycf3) has two introns. The phylogenomic analysis reveals that D. hamiltonii clustered together with D. brandisii (Dendrocalamus that constituted a monophyletic, and was sister to Bambusa within Bambusoideae.The cp genome of randisii . Five sa