Extracellular vesicles(EVs)are nano-sized bilayer vesicles that are shed or secreted by virtually every cell type.A variety of biomolecules,including proteins,lipids,coding and non-coding RNAs,and mitochondrial DNA,can be selectively encapsulated into EVs and delivered to nearby and distant recipient cells,leading to alterations in the recipient cells,suggesting that EVs play an important role in intercellular communication.EVs play effective roles in physiology and pathology and could be used as diagnostic and therapeutic tools.At present,although the mechanisms of exosome biogenesis and secretion in donor cells are well understood,the molecular mechanism of EV recognition and uptake by recipient cells is still unclear.This review summarizes the current understanding of the molecular mechanisms of EVs'biological journey in recipient cells,from recognition to uptake and cargo release.Furthermore,we highlight how EVs escape endolysosomal degradation after uptake and thus release cargo,which is crucial for studies applying EVs as drug-targeted delivery vehicles.Knowledge of the cellular processes that govern EV uptake is important to shed light on the functions of EVs as well as on related clinical applications.

Objective:To train a deep convolutional neural networks (CNN) using a labeled data set to classify the metaphase chromosomes and test its accuracy for chromosomal identification.Methods:Three thousand and three hundred individuals undergoing surveillance for chromosomal disorders at the Laboratory for Comprehensive Prevention and Treatment of Birth Defects, Ningbo Maternal and Child Health Care Hospital from January 2013 to July 2019 were enrolled. A total of 3 300×46 chromosome images were included, of which 70% were used as the training set and 30% were used as the test set for the deep CNN. The accuracy of chromosome counting and "cutting + recognition + arrangement + automatic analysis" of the model were respectively evaluated. Another 80 images were collected to record the time and accuracy of chromosome classification by geneticists and the model, respectively, so as to assess the practical value of the model.Results:The CNN model was used to count the chromosomes with an accuracy of 61.81%, and the "cutting + recognition + arrangement + automatic analysis" accuracy of the model was 96.16%. Compared with manual operation, the classification time of the CNN model has been greatly reduced, and its karyotyping accuracy was only 3.58% lower than that of geneticists.Conclusion:The CNN model has a high performance for chromosome classification and can significantly reduce the work load involved with the segmentation and classification and improve the efficiency of chromosomal karyotyping, thereby has a broad application prospect.

Objective To explore the changes of resting-state degree centrality(DC)in elderly patients with chronic insomnia disorder(CID).Methods Resting-state functional magnetic resonance imaging(rs-fMRI)data were collected from 26 untreated elderly patients with CID(CID group)and 45 healthy controls(HC)(HC group).Two-sample t-test was conducted to compare the intergroup differences in whole-brain DC values,and the correlation between DC values in different brain regions and clinical indicators were analyzed,and logistic regression analysis was performed to verify the diagnostic efficacy of changes in DC values for elderly CID.Results Compared with the HC group,the DC values of the right insula,left rolandic operculum,and opercular part of right inferior frontal gyrus in the elderly CID group decreased[P＜0.05,false discovery rate(FDR)corrected],while the DC values of the right middle frontal gyrus increased(P＜0.05,FDR corrected).And the DC values of the opercular part of right inferior frontal gyrus in the elderly CID group were positively correlated with sleep efficiency(r=0.504,P=0.009)and self-rating depression scale(SDS)(r=0.401,P=0.042),respectively.The sensitivity of DC value in the opercular part of right inferior frontal gyrus for diagnosing elderly CID was 0.822,the specificity was 0.615,and the accuracy was 0.701.Conclusion Elderly CID patients have abnormal DC values in the right insula,left rolandic operculum,opercular part of right inferior frontal gyrus and right middle frontal gyrus,which may provide imaging evidence for exploring the pathogenesis of CID and clinical diagnosis and treatment.

Objective To develop and validate a risk prediction model based on early platelet-related parameters for bronchopulmonary dysplasia(BPD)in neonates admitted to the neonatal intensive care unit(NICU),and to facilitate early identification and intervention in high-risk populations.Methods Clinical data of 291 preterm infants with a gestational age(GA)≤32 weeks or a birth weight(BW)＜1 500 g,admitted to the NICU,were retrospectively analyzed.Out of these,214 cases were selected as the modeling group.This group was further categorized into the BPD group(n=76)and the non-BPD group(n=138),based on whether they required oxygen therapy at 28 days post-birth.Perinatal data,platelet-related parameters and other indicators between the two groups.Univariate and multivariate Logistic regression analyses were conducted to identify BPD risk factors,followed by the construction of a nomogram.An additional cohort of 105 preterm infants with GA≤32 weeks or BW＜1 500 g,were used to validate the model.This cohort was divided into the BPD group(n=43)and the non-BPD(n=62)group.Receiver operating characteristic(ROC)curve and calibration curve were used to internally verify the efficiency of the prediction model.Results The Logistic regression analysis identified GA,BW,Apgar score at 5 minutes≤7,invasive ventilation,platelet count(PLT)and mean platelet volume(MPV)as significant factors in the model(P＜0.05).The constructed nomogram was formulated using R language,and the areas under the ROC curve(AUC)for the three models were 0.908,0.931 and 0.918,respectively(P＜0.05).The verification group was verified by Bootstrap.The calibration curve showed a good fit.The internal validation AUC values of the three models were 0.877,0.890 and 0.886,respectively.Conclusion GA,BW,invasive ventilation,Apgar score at 5 minutes≤7,MPV and PLT are key risk factors for BPD onset.The risk prediction model based on these indicators can effectively predict BPD,providing clinicians with a valuable tool for early detection and intervention in the development of BPD.

Objective:To investigate the correlation of peripheral blood relative mitochondrial DNA copy number(mtDNAcn)with intrinsic capacity and body composition, and to identify potential biomarkers for healthy aging.Methods:Clinical data of 416 patients admitted to our hospital from September 2019 to June 2021 were consecutively collected.MtDNA was extracted from peripheral blood of these subjects, and mtDNAcn was determined by a real-time fluoresence quantitative reverse transcription-polymerase chain reaction(qRT-PCR). Intrinsic capacity assessment included 5 aspects that were exercise[Morse Fall Scale(MFS), Physiological Frailty Phenotype(PFP), Sarcopenia Questionnaire(SARC-CALF), Short Physical Performance Battery(SPPB), Time Up and Go Test(TUG)]; vitality[Mini Nutritional Assessment(MNA), Multidimensional Prognostic Index(MPI)]; cognition[Mini-Mental State Examination(MMSE)scale]; psychology[Geriatric Depression Scale(GDS), Self-rating Anxiety Scale(SAS)]; sensory capacities[Cumulative Illness Rating Scale-the Comorbidity Index(CIRS-CI)]. To assess body composition, dual-energy X-ray absorptiometry was used to measure body fat, including trunk fat, total body fat, fat in the abdominal region, fat in the buttock region, and then to calculate fat index(FMI)and limb skeletal muscle mass index(ASMI).Results:Spearman correlation analysis showed that mtDNAcn had a negatively correlation with age( r=-0.176, P<0.05). After adjustment for gender and body mass index, partial correlation analysis showed mtDNAcn were still negatively correlated with age( r=-0.144, P<0.05). Furthermore, mtDNAcn was significantly correlated with 4 m gait speed, the scores of SARC-CalF, MFS, MNA, MMSE, MPI and its sub-scale's Activities of Daily Living(ADL)and Short Portable Mental Status Questionnaire(SPMSQ)( r=0.171, -0.207, -0.163, 0.221, 0.184, -0.210, 0.241, -0.269, all P<0.05). After adjustment for age, gender and body mass index, partial correlation analysis showed mtDNAcn still had a significant correlation with gait speed, the scores of MFS, MNA, MPI and SPMSQ( r=0.170, -0.170, 0.148, -0.242, -0.188, all P<0.05). In addition, the Spearman correlation analysis showed that mtDNAcn was positively correlated with FMI, trunk fat, total body fat, abdominal fat and fat in the buttock region( r=0.168, 0.143, 0.175, 0.116, 0.199, all P<0.05). However, after adjustment for age and gender, mtDNAcn was only correlated with FMI, total body fat, fat in the buttock region( r=0.126, 0.131, 0.127, all P<0.05). On the other hand, multiple linear regression analysis showed that mtDNAcn was significantly correlated with age, gait speed, FMI, total body fat, fat in the buttock region, the scores of MFS, PFP, MNA and MPI( β=-0.191, 0.156, 0.126, 0.131, 0.125, -0.119, -0.145, 0.151, -0.171, all P<0.05). Conclusions:MtDNAcn is correlated with physical function, frailty, nutrition, falling, cognition and body composition, and may be considered as a biomarker for the evaluation of the locomotion and vitality of human intrinsic capacity.

Objective:To establish and validate an LC-MS/MS method for simultaneous determination of Aβ 1-42, Aβ 1-40, and Aβ 1-38 in cerebrospinal fluid. Additionally, the consistency between this method and three mainstream detection methods was evaluated.Methods:This study involved method establishment, validation, and consistency evaluation. The N15 labeled β-amyloid protein was used as the internal standard. Extraction was performed using Waters MCX 96-wells solid phase extraction plate, and the eluent was collected to QuanRecovery MaxPeak 700 μl plate. At the positive ion mode, the multi-reaction ion monitoring mode based on electric spray ionization is chosen for the determination of CSF Aβ 1-42, Aβ 1-40, and Aβ 1-38. Referring to the CLSI C62-A and EP-15A3 guidelines, the method is evaluated and verified, including quantitation of limit (LOQ), linearity, recovery, precision, and accuracy. In addition, a total of 57 clinical residual CSF samples were collected and the concentrations of Aβ 1-42 and Aβ 1-40 were determined based on manual INNOTEST ELISA assay and Lumipulse G and Roche Elecsys fully automated biochemical analyzers. The comparison analysis and deviation evaluation were conducted by passing-bablok and Bland Altman methods.Results:The analysis time of this method is 8 min, and the LOQ of Aβ 1-42, Aβ1-40 and Aβ1-38 is 0.1 ng/ml, 0.5 ng/ml, and 0.1 ng/ml, respectively, and the linear range can meet the needs of clinical detection. Respectively, the recovery is 86.2%-93.8%, 100.9%-103.9% and 103.3%-107.1%; the total imprecision is 4.7%-7.4%, 3.5%-4.6% and 5.2%-10.9%. The measured values of Aβ 1-42 certified reference materials are all within the allowable uncertainty requirements. Moreover, the carryover rate of three analytes was all≤0.11%. In addition, the correlations of Aβ 1-42 and Aβ1-40 in CSF between this LC-MS/MS method and the INNOTEST ELISA method, Lumipulse G and Roche Elecsys fully automated biochemical analyzers were all deemed good, with correlation coefficient (r) ranging from 0.920 to 0.970. However, the measured values between the four methods were remarkably different.Conclusion:We established and validated a robust method based on LC-MS/MS technology for simultaneous determination of Aβ 1-42, Aβ 1-40, and Aβ 1-38 in CSF. The method is accurate, simple, and suitable for clinical measurements. However, despite good correlations, there were substantial differences in the measurement results of Aβ 1-42 and Aβ 1-40 among different analytical platforms, indicating the need for further promotion of harmonization and standardization processes for AD classic biomarkers.

Objective:To investigate the application value of high-resolution CT in the diagnosis of early lung adenocarcinoma.Methods:The clinical data of 90 patients with early lung adenocarcinoma who received treatment from January to December 2021 in Quzhou People's Hospital were retrospectively analyzed. These patients were divided into a non-invasive group ( n = 27) and an invasive group ( n = 63) according to whether they had invasive lung adenocarcinoma or not. In the non-invasive group, there were 10 patients with atypical adenomatous hyperplasia and 17 patients with in situ adenocarcinoma. In the invasive group, there were 38 patients with minimally invasive adenocarcinoma and 25 patients with invasive adenocarcinoma. A pathological diagnosis was performed in all patients. All patients underwent a high-resolution CT examination. The incidence of lung adenocarcinoma was compared between different loci. CT signs were compared between the non-invasive group and the invasive group. The receiver operating characteristic curve was used to determine the sensitivity and specificity of high-resolution CT in the differential diagnosis of lung adenocarcinoma prior to invasion. Results:There was no significant difference in the incidence of lung adenocarcinoma among the left upper lobe, right upper lobe, left lower lobe, and right lower lobe of the lung in each group (all P > 0.05). The detection rates of pleural indentation sign, spicule sign, and vascular bundle sign in the invasive group were 28.6% (18/63), 31.57% (20/63), and 27.0% (17/63), respectively, which were significantly higher than 7.41% (2/27), 11.11% (3/27), and 7.41% (2/27) in the non-invasive group ( χ2 = 4.90, 4.23, 4.35, all P < 0.05). There were no significant differences in detection rates in the lobar sign and air bronchogram sign between the two groups (both P > 0.05). The receiver operating characteristic curve analysis results showed that the sensitivity and specificity of high-resolution CT in differentiating pre-invasive lesions were 100.0% and 64.7%, respectively. Conclusion:High-resolution CT has a good diagnostic value for early lung adenocarcinoma. It can clearly display various signs of early lung adenocarcinoma, which is worthy of clinical reference.

Graph-theory-based pathway analysis is a commonly used method for pathway searching in genome-scale metabolic networks. However, such searching often results in many pathways biologically infeasible due to the presence of currency metabolites (e.g. H⁺, H₂O, CO₂, ATP etc.). Several methods have been proposed to address the problem but up to now there is no well-recognized methods for processing the currency metabolites. In this study, we proposed a new method based on the function of currency metabolites for transferring of functional groups such as phosphate. We processed most currency metabolites as pairs rather than individual metabolites, and ranked the pairs based on their importance in transferring functional groups, in order to make sure at least one main metabolite link exists for any reaction. The whole process can be done automatically by programming. Comparison with existing approaches indicates that more biologically infeasible pathways were removed by our method and the calculated pathways were more reliable, which may facilitate the graph-theory-based pathway design and visualization.
Genome ; Metabolic Networks and Pathways

Objective:To compare the difference of epileptiform discharges detection in patients with epilepsy between the 25 electrodes electroencephalogram (EEG) system proposed by the International Federation of Clinical Neurophysiology in 2017 and the previous 19 electrodes EEG system.Methods:Patients suspected of epilepsy or with confirmed epilepsy who need a follow-up EEG were collected in Zhongshan Hospital, Fudan University from March 2018 to November 2019, and conventional video-EEG recording was performed on all patients for two hours with the standard 25 electrodes EEG system. Two neurophysiologists reviewed the recordings blindly using the 19 electrodes system and the 25 electrodes system, marking the epileptiform discharges and their amplitudes. Finally, the data were statistically analyzed.Results:A total of 403 patients were included in the study, in which 263 cases were diagnosed as epilepsy, including 129 cases of generalized epilepsy, 115 cases of temporal lobe epilepsy, 13 cases of frontal lobe epilepsy, two cases of parietal lobe epilepsy and four cases of occipital lobe epilepsy. In 115 temporal lobe epilepsy patients, 76 (66.09%) and 100 (86.96%) records were detected epileptiform discharges by the 19 or 25 electrodes EEG system respectively, and the difference was statistically significant (χ2=13.939, P<0.001). While in patients with non-temporal lobe epilepsy, there was not statistically significant difference between the two systems. In 76 patients whose temporal epileptiform discharges were detected by the two systems, the amplitudes of epileptiform discharges in the newly-added inferior temporal electrodes (F9/F10, T9/T10, P9/P10) and the original temporal electrodes (F7/F8, T7/T8, P7/P8) were (61.53±22.64) μV and (48.25±20.90) μV, respectively, with statistically significant difference between the two groups ( t=5.486, P<0.001). In patients with abnormal [79.59% (39/49) vs 61.22% (30/49), χ2=3.967, P=0.046] and normal [95.45% (42/44) vs 70.45% (31/44), χ2=9.724, P=0.003] imaging, the ability of the 25 electrodes EEG system to detect epileptiform discharges was higher than that of the 19 electrodes EEG system. Conclusion:The 25 electrodes EEG system can significantly improve the detection ability of temporal epileptiform discharges in patients with epilepsy, which is recommended for regular use to increase the detection ability of temporal area abnormal wave and assist the diagnosis and treatment of epilepsy.

The traditional paradigm of motor-imagery-based brain-computer interface (BCI) is abstract, which cannot effectively guide users to modulate brain activity, thus limiting the activation degree of the sensorimotor cortex. It was found that the motor imagery task of Chinese characters writing was better accepted by users and helped guide them to modulate their sensorimotor rhythms. However, different Chinese characters have different writing complexity (number of strokes), and the effect of motor imagery tasks of Chinese characters with different writing complexity on the performance of motor-imagery-based BCI is still unclear. In this paper, a total of 12 healthy subjects were recruited for studying the effects of motor imagery tasks of Chinese characters with two different writing complexity (5 and 10 strokes) on the performance of motor-imagery-based BCI. The experimental results showed that, compared with Chinese characters with 5 strokes, motor imagery task of Chinese characters writing with 10 strokes obtained stronger sensorimotor rhythm and better recognition performance (
Brain-Computer Interfaces ; China ; Electroencephalography ; Evoked Potentials ; Humans ; Imagery, Psychotherapy ; Imagination