Objective To analyze the composition characteristics and biological functions of tumor cell subpopulations in glioblastoma(GBM)through multi-transcriptomics sequencing technology,and explore the hypoxia characteristics and spatial localization features of the mesenchymal-like(MES-like)tumor cell subpopulation in GBM and the influence on malignant biological behaviors.Methods Multi-transcriptomics sequencing data,including single-cell RNA sequencing(scRNA-seq)data(18 patients),bulk RNA sequencing(bulk RNA-seq)and spatial transcriptomics(ST)data of GBM,were employed to define cell subpopulations in GBM,and Gene Ontology(GO)and Gene Set Enrichment Analysis(GSEA)were utilized to analyze their functions.The proportions and locations of cell subpopulations in bulk RNA-seq data were evaluated with BayesPrism deconvolution.Immunofluorescence assay was conducted for verification on 12 paraffin samples of GBM from patients who visited the neurosurgical department of our hospital from 2015 to 2023 and met the pathological diagnostic criteria for GBM(10 males and 2 females,at an average age of 53.50 years and a median age of 54.50 years).pySCENIC was applied to predict specific transcription factors of tumor cell subpopulations.Results Tumor cells in GBM were highly heterogeneous,and could be mainly divided into 4 subpopulations:astrocyte-like(AC-like),neural progenitor-like(NPC-like),oligodendrocyte progenitor-like(OPC-like)and MES-like.Differential gene analysis found that the MES-like tumor cells highly expressed vascular endothelial growth factor A(VEGFA),adrenomedullin(ADM),N-myc downstream regulated 1(NDRG1),insulin like growth factor binding protein 5(IGFBP5),and A-kinase anchoring protein 12(AKAP12)(P<0.001).pySCENIC transcription factor prediction found that the high-active transcription factors of the MES-like tumor cells were AT-rich interaction domain 3A(ARID3A),FOS like 2,AP-1 transcription factor subunit(FOSL2),endothelial PAS domain protein 1(EPAS1),CCAAT enhancer binding protein delta(CEBPD),and CCAAT enhancer binding protein beta(CEBPB)(P<0.05).GO and GSEA enrichment analyses found that the MES-like tumor cells were enriched in hypoxia-related pathways,especially the pathway of cell responses to hypoxia levels(NES=2.437,P<0.001).BayesPrism deconvolution showed that the MES-like tumor cells mainly existed in PAN(Pseudopalisading cells around necrosis)and perinecrotic zone.Immunofluorescence assay confirmed CD44+(CD44 antigen)MES-like tumor cells were mainly located in hypoxia areas with highly expression of hypoxia inducible factor 1 subunit alpha(HIF1α)(P<0.01).Multivariate Cox regression analysis indicated that the MES-like tumor cells were significantly correlated with the adverse prognosis of GBM patients(HR=1.71,95%CI:1.38～2.11,P<0.001).Conclusion Tumor cells in GBM are of highly heterogeneity.They could be mainly divided into 4 subpopulations:AC-like,NPC-like,OPC-like and MES-like.MES-like tumor cells,mainly locating in PAN and perinecrotic zone,are characterized by hypoxia,which can promote the malignant progression of GBM.

Depression after myocardial infarction is closely related to the theory of ascending and descending of qi and blood. The core pathogenesis is analyzed as disorder of qi and blood, mental damage, uncontrolled upward and downward movement, and pivot movement failure. The treatment method is to regulate qi and blood, invigorate qi and activate blood circulation and restore the rise and fall of visceral qi. Nourishing qi and promoting blood circulation should invigorate qi in the first. Appropriate blood activating drugs should be selected according to the degree of blood stasis, so that the blood circulation is smooth and the spirit has dependence. The key to restoring the normal balance of qi movement of visceral organs is to regulate not only liver ascending and lung descending but also spleen. Clinicians need pay attention to the nature of drug lifting and falling, conforming the physiological functions of the organs, to restore the rising and falling of the qi movement of internal organs.

This study aims to investigate the effects of Japanese encephalitis virus(JEV)on TLRs signaling pathway and its regulation of the secretion of inflammatory factors during the infection of testicular interstitial cells,In this study,the mRNA levels of TLR3,TLR7,TLR8,TRIF and MyD88 genes were detected by qPCR after 1 MOI dose of JEV was inoculated into testicular stro-mal cells at different time periods.Western blot assay was used to detect the expression levels of TLR3,TLR7,TRIF and MyD88 protein at 6 h after JEV infection,and ELISA was used to detect the expression levels of IL-1β,IL-6 and TNF-α at different time periods(6,12 and 24 h).The re-sults showed as follows:After 6 h of JEV infection,the mRNA levels of TLR3,TLR7,TRIF and MyD88 genes were significantly up-regulated(P＜0.05),and the mRNA levels of TLR8 genes were down-regulated(P＜0.05).Western blot results showed that the protein expressions of TLR3,TLR7,TRIF and MyD88 were significantly up-regulated when JEV infected testicular stromal cells for 6 h(P＜0.05),which was consistent with the corresponding mRNA transcription levels.There was no significant change in TLR8 protein expression.ELISA results showed that 6 h after JEV infection of testicular stromal cells,IL-6 was significantly increased(P＜0.01),and the expressions of IL-1β and TNF-α were not changed.TLR3,TLR7,TLR8,TRIF and MyD88 were si-lenced by siRNA,and the silenced cells were inoculated with JEV for 6 h,and IL-6 expression lev-els were detected by ELISA.The results showed that silenced TLR3,TLR7,TLR8,TRIF and MyD88 could significantly reduce the increase of IL-6 secretion induced by JEV infection(P＜0.05).These results indicated that JEV could induce the expression of inflammatory factor IL-6 by activating TLR3,TLR7 and TLR8 signaling pathway after infection of testicular stromal cells.This study provides a reference for further elucidating the mechanism of reproductive disorders caused by JEV infection.

Porcine circovirus type 2(PCV2)is the main pathogen causing porcine circovirus related diseases.PCV2 infection in pigs may lead to porcine dermatitis and nephrotic syndrome(PDNS)and weaned piglets multiple system failure syndrome(PMWS),etc.At present,the pathogenic mechanism is not fully understood.PCV2 is a single strand of negative link DNA,which can cause immune suppression in the body and lead to increased secondary susceptibility,which has a syner-gistic effect with various pig diseases and brings major economic losses to the pig industry.Al-though there are commercial vaccines,the prevention of vaccines has certain limitations and there is no effective drug treatment so far,an outbreak will threaten people's life and health and public safety,resulting in significant economic losses.In order to understand the latest progress of PCV2 escape mechanism and prevention and control,this paper summarizes the inhibition of interferon production,regulation of apoptosis,regulation of autophagy,regulation of pyroptosis and inflam-matory response,evasion of adaptive immune response,and prevention and control of PCV2,in or-der to provide new theoretical ideas for the research and prevention and control of PCV2.

Axenfeld-Rieger syndrome is a rare autosomal dominant hereditary disease characterized by anterior segment dysgenesis, which may be accompanied by various systemic defects, including craniofacial dysmorphism, hypodontia, microdontia, and redundant periumbilical skin.Its typical ocular manifestations include posterior embryotoxon, iris hypoplasia, peripheral anterior synechiae, corectopia and polycoria with a high prevalence of glaucoma.Patients can exhibit any combination of these features.However, family members with the same genotype may present different phenotypes due to phenotypic heterogeneity.Emerging evidence suggests that PITX2 and FOXC1 genes encoding transcription factors are primarily associated with genetic variants in ARS.Intragenic mutations and gene deletions are common types of genetic variations suspected to trigger changes in gene dosages and protein function.However, the underlying molecular mechanism remains unclear.Some patients with ARS carry mutations in the COL4A1, PRDM5, and CYP1B1 genes, but the pathogenicity of these variations has yet to be confirmed by further studies.This article provided an overview of the typical clinical features, potential correlations between phenotype and genotype, as well as gene function.

In recent years, wearable devices have seen a booming development, and the integration of wearable devices with clinical settings is an important direction in the development of wearable devices. The purpose of this study is to establish a prediction model for postoperative pulmonary complications (PPCs) by continuously monitoring respiratory physiological parameters of cardiac valve surgery patients during the preoperative 6-Minute Walk Test (6MWT) with a wearable device. By enrolling 53 patients with cardiac valve diseases in the Department of Cardiovascular Surgery, West China Hospital, Sichuan University, the grouping was based on the presence or absence of PPCs in the postoperative period. The 6MWT continuous respiratory physiological parameters collected by the SensEcho wearable device were analyzed, and the group differences in respiratory parameters and oxygen saturation parameters were calculated, and a prediction model was constructed. The results showed that continuous monitoring of respiratory physiological parameters in 6MWT using a wearable device had a better predictive trend for PPCs in cardiac valve surgery patients, providing a novel reference model for integrating wearable devices with the clinic.
Humans ; Lung ; Walking/physiology* ; Walk Test ; Heart Valves/surgery* ; Postoperative Period ; Postoperative Complications/etiology*

Objective:To identify disease-causing variation in a Chinese family with Axenfeld-Rieger syndrome (ARS) through the analysis of clinical symptoms and hereditary information.Methods:The method of pedigree investigation was adopted.A Chinese ARS family including 15 family members of 3 generations was recruited in the Second Affiliated Hospital of Harbin Medical University in 2018.There were 3 patients in the family.The family history and clinical data were collected.Ophthalmic and general examinations were carried out in all the members included.DNA and RNA were extracted from collected peripheral venous blood samples of 2-5 ml from each member.Whole exome sequencing was used to screen the variations in the proband.Suspected variations screened through searching population databases and bioinformatics analysis were verified by Sanger sequencing and real-time quantitative PCR.Conservation analysis and deleteriousness prediction of suspected variations were conducted.The pathogenecity of candidate rare variations were evaluated according to the American College of Medical Genetics and Genomics (ACMG) standards and guidelines.This study followed the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the Second Affiliated Hospital of Harbin Medical University (No.KY2019-231).Written informed consent was obtained from each subject or custodian prior to entering the study cohort.Results:The 3 patients all had typical ARS clinical features in eyes, teeth and umbilicus, and carried the same heterozygous variant, c.525delC (p.Asp175Glufs *) in the PITX2 gene, which were not found in other members, indicating co-segregation.The relative expression of PITX2 mRNA was 0.672±0.063 in the patients, which was significantly lower than 1.015±0.179 in the healthy controls ( t=8.847, P<0.001).This variant was not recorded in dbSNP, 1000G, gnomeAD, ExAC, Korea1K and EVS databases, and it was labelled as deleterious by MutationTaster.The affected conservative amino acid sequences were found in 9 species.The variant was determined as pathogenic according to the ACMG standards and guidelines. Conclusions:The c.525delC (p.Asp175Glufs *) mutation of PITX2 gene is pathogenic in the pedigree.This is the first time that this mutation has been reported in Chinese family with ARS.

Breathing pattern parameters refer to the characteristic pattern parameters of respiratory movements, including the breathing amplitude and cycle, chest and abdomen contribution, coordination, etc. It is of great importance to analyze the breathing pattern parameters quantificationally when exploring the pathophysiological variations of breathing and providing instructions on pulmonary rehabilitation training. Our study provided detailed method to quantify breathing pattern parameters including respiratory rate, inspiratory time, expiratory time, inspiratory time proportion, tidal volume, chest respiratory contribution ratio, thoracoabdominal phase difference and peak inspiratory flow. We also brought in "respiratory signal quality index" to deal with the quality evaluation and quantification analysis of long-term thoracic-abdominal respiratory movement signal recorded, and proposed the way of analyzing the variance of breathing pattern parameters. On this basis, we collected chest and abdomen respiratory movement signals in 23 chronic obstructive pulmonary disease (COPD) patients and 22 normal pulmonary function subjects under spontaneous state in a 15 minute-interval using portable cardio-pulmonary monitoring system. We then quantified subjects' breathing pattern parameters and variability. The results showed great difference between the COPD patients and the controls in terms of respiratory rate, inspiratory time, expiratory time, thoracoabdominal phase difference and peak inspiratory flow. COPD patients also showed greater variance of breathing pattern parameters than the controls, and unsynchronized thoracic-abdominal movements were even observed among several patients. Therefore, the quantification and analyzing method of breathing pattern parameters based on the portable cardiopulmonary parameters monitoring system might assist the diagnosis and assessment of respiratory system diseases and hopefully provide new parameters and indexes for monitoring the physical status of patients with cardiopulmonary disease.
Humans ; Lung ; Pulmonary Disease, Chronic Obstructive ; Respiration ; Tidal Volume ; Wearable Electronic Devices

As a low-load physiological monitoring technology, wearable devices can provide new methods for monitoring, evaluating and managing chronic diseases, which is a direction for the future development of monitoring technology. However, as a new type of monitoring technology, its clinical application mode and value are still unclear and need to be further explored. In this study, a central monitoring system based on wearable devices was built in the general ward (non-ICU ward) of PLA General Hospital, the value points of clinical application of wearable physiological monitoring technology were analyzed, and the system was combined with the treatment process and applied to clinical monitoring. The system is able to effectively collect data such as electrocardiogram, respiration, blood oxygen, pulse rate, and body position/movement to achieve real-time monitoring, prediction and early warning, and condition assessment. And since its operation from March 2018, 1 268 people (657 patients) have undergone wearable continuous physiological monitoring until January 2020, with data from a total of 1 198 people (632 cases) screened for signals through signal quality algorithms and manual interpretation were available for analysis, accounting for 94.48 % (96.19%) of the total. Through continuous physiological data analysis and manual correction, sleep apnea event, nocturnal hypoxemia, tachycardia, and ventricular premature beats were detected in 232 (36.65%), 58 (9.16%), 30 (4.74%), and 42 (6.64%) of the total patients, while the number of these abnormal events recorded in the archives was 4 (0.63%), 0 (0.00%), 24 (3.80%), and 15 (2.37%) cases. The statistical analysis of sleep apnea event outcomes revealed that patients with chronic diseases were more likely to have sleep apnea events than healthy individuals, and the incidence was higher in men (62.93%) than in women (37.07%). The results indicate that wearable physiological monitoring technology can provide a new monitoring mode for inpatients, capturing more abnormal events and provide richer information for clinical diagnosis and treatment through continuous physiological parameter analysis, and can be effectively integrated into existing medical processes. We will continue to explore the applicability of this new monitoring mode in different clinical scenarios to further enrich the clinical application of wearable technology and provide richer tools and methods for the monitoring, evaluation and management of chronic diseases.
Heart Rate ; Humans ; Monitoring, Physiologic ; Movement ; Sleep Apnea Syndromes ; Wearable Electronic Devices

Objective:To investigate the effect of sevoflurane anesthesia on electroencephalographic (EEG) seizures and long-term behavior and possible mechanism in neonatal rats.Methods:A total of 141 postnatal days 4-6 Sprague-Dawley rats (66 male, 75 female) were divided into 3 groups ( n=47 in each group) according to random number table method: control group, sevoflurane group, and NKCC1 inhibitor group, with 22 males and 25 females in each group. Rats in the control group were fed in normal cage without anesthesia; rats in the sevoflurane group were anesthetized with 2.1% sevoflurane for 6 hours; rats in the NKCC1 blocker group received intraperitoneal injection of 1.82 mg / kg bumetanide 30 minutes before anesthesia with 2.1% sevoflurane. The rats in the control group and sevoflurane group were injected subcutaneously with the same dose of DMSO at the same time when the NKCC1 blocker group received the drug intervention, so as to eliminate the influence caused by the solvent. The rats were observed for 30 minutes after recovery from anesthesia and then continued to breastfeed normally. Some of the new born rats received EEG monitoring from 9 to 11 days after being raised; the other rats received EPM and PPI respectively at 60 and 70 days after being raised. Results:The results of EEG showed that, compared with the control group, the number of epileptic waves((0.429±0.787), (1.571±0.787), t=2.753, P<0.01), the average duration of single epileptic wave ((1.575±2.349), (6.392±3.374), t=3.880, P< 0.01), the total duration increased significantly ((1.800±3.617), (10.957±6.028), t= 3.929, P<0.01) were all increased, the differences were statistically significant. Compared with sevoflurane group, the number of epileptic waves in EEG of male rats in NKCC1 blocker group decreased, the average duration of single epileptic wave decreased, and the total duration of epileptic wave shortened significantly, with statistical significance ((0.286±0.756), (0.925±1.733), (1.043±2.759), t=3.097, 4.404, 4.254, all P<0.01). There were no significant differences in female rat among the three groups (all P>0.05). Compared with male rats, the average duration of female rats in sevoflurane group decreased ((6.392±3.374), (2.515±2.992), t=3.044, P<0.01), the total duration shortened ((10.957±6.028), (3.270±5.883), t=2.626, P<0.01), the difference was statistically significant.The behavioral results showed that, compared with the control group, the open arm dwell time of male rats in sevoflurane group was significantly shorter ( P<0.05), and the panic response in PPI group was significantly lower ( P<0.05), the difference was statistically significant.Compared with the sevoflurane group, the open arm dwell time in NKCC1 blocker group was significantly longer ( P<0.05), and the panic response in PPI group was significantly increased.The difference was statistically significant ( P<0.05). The change trend in female rats of each group was similar to that of male rats, but there was no significant difference (all P>0.05). Comparison between male and female rats: compared with male rats in sevoflurane group, the female rats in sevoflurane group had a longer open arm stay time in EPM experiment ( P<0.05), the difference was statistically significant. Conclusion:Sevoflurane anesthesia for 6 hours can significantly increase the generation of epileptic waves in EEG of male newborn rats, and cause behavioral abnormalities in adult male rats, which may be related to NKCC1.And male rats are more vulnerable to the negative effects of sevoflurane anesthesia on brain nerve development.