Extensive epigenetic reprogramming involves in memory CD8+ T-cell differentiation. The elaborate epigenetic rewiring underlying the heterogeneous functional states of CD8+ T cells remains hidden. Here, we profile single-cell chromatin accessibility and map enhancer-promoter interactomes to characterize the differentiation trajectory of memory CD8+ T cells. We reveal that under distinct epigenetic regulations, the early activated CD8+ T cells divergently originated for short-lived effector and memory precursor effector cells. We also uncover a defined epigenetic rewiring leading to the conversion from effector memory to central memory cells during memory formation. Additionally, we illustrate chromatin regulatory mechanisms underlying long-lasting versus transient transcription regulation during memory differentiation. Finally, we confirm the essential roles of Sox4 and Nrf2 in developing memory precursor effector and effector memory cells, respectively, and validate cell state-specific enhancers in regulating Il7r using CRISPR-Cas9. Our data pave the way for understanding the mechanism underlying epigenetic memory formation in CD8+ T-cell differentiation.
CD8-Positive T-Lymphocytes/metabolism* ; Cell Differentiation ; Chromatin/immunology* ; Animals ; Mice ; Immunologic Memory ; Epigenesis, Genetic ; SOXC Transcription Factors/immunology* ; NF-E2-Related Factor 2/immunology* ; Mice, Inbred C57BL ; Gene Regulatory Networks ; Enhancer Elements, Genetic

Fetal structural anomalies and birth defects are primarily caused by genetic variants such as chromosomal number abnormalities, copy number variations (CNV), single nucleotide variants (SNV), and small insertions and deletions (indel). Whole-genome sequencing (WGS) based on next-generation sequencing (NGS) as an emerging technology for genetic disease diagnosis can detect the aforementioned types of variants. In recent years, high-depth WGS (> 30×) for prenatal diagnosis has also become available, and proved to be practical for unraveling the genetic etiology of fetal developmental abnormalities. To fascilitate clinical practice, test development and preliminary implementation of WGS for diagnosing fetal structural anomalies, we have formulated a consensus over the application of WGS in prenatal diagnosis by compiling previously published consensuses, guidelines, and research findings to provide a guidance on data analysis, reporting recommendations, and consultation of prenatal WGS results.

INTRODUCTION: Timely detection of dementia enables early access to dementia-specific care services and interventions. Various stakeholders brought together to refine Singapore's dementia care strategy identified a lack of a standardised cognitive screening tool and the absence of a comparative review of existing tools. We hence conducted a rapid review to evaluate the diagnostic performance of brief cognitive screening tools in identifying possible dementia among community-dwelling older adults in Singapore. METHOD: Brief cognitive screening tools were defined as interviews or tests administered in ≤5 minutes. Studies performed in Singapore on older adults ≥60 years, which used locally-validated comparators and reported outcomes of clinician-diagnosed dementia were included. Rapid review methodology was used in study screening and selection. Quality Assessment of Diagnostic Accuracy Studies version 2 tool was used for risk-of-bias assessment. A negative likelihood ratio (LR-) of ≤0.2 was defined a priori as having a moderate effect in shifting post-test probability. RESULTS: Fourteen studies were included in qualitative synthesis: 3 studies evaluated self-/informant-based tools only, 4 evaluated performance-based measures only and 7 evaluated combination approaches. Eight-item Informant Interview to Differentiate Aging and Dementia (AD8) was the most studied self-/ informant-based tool. One study found informant AD8 (iAD8) superior to self-rated AD8. Another study found iAD8 superior to Mini-Mental State Examination. Among performance-based measures, Abbreviated Mental Test, Visual Cognitive Assessment Test-Short form version 1 (VCAT-S1), VCAT-S2 and Mini-Cog had LR- <0.2. Minimal improvement of combination approaches compared to iAD8 alone was demonstrated. CONCLUSION Our review suggests the limited utility of dementia screening in communities with low dementia prevalence and supports a case-finding approach instead. With a reliable informant, iAD8 alone has sufficient discriminant ability. Further research is needed to specifically assess the diagnostic ability of performance-based tools in community settings.
Humans ; Singapore ; Dementia/diagnosis* ; Aged ; Independent Living ; Mass Screening/methods* ; Middle Aged ; Aged, 80 and over

Objective To investigate the shedding of CAV2-ΔE3-CGS after immunization and the background of canine adenovirus (CAV) infection, and to establish a dual fluorescent quantitative PCR detection method for rabies virus (RV) and canine adenovirus type 2 (CAV2). Methods A dual fluorescent quantitative PCR detection method was established by designing specific primers and probes for E1 gene of CAV and G gene of RV for the detection of CAV2-ΔE3-CGS. Oral swabs, anal swabs and environmental samples of stray dogs from experimental animal farm and dog detention center were tested. Results The standard curves generated by this method were Y=-3.351 × logX + 44.895, R² = .999 and Y=-3.413 × logX + 45.192, R²=0.996, respectively. The linear relationships were good, and the minimum detection limits were both 102 copies/μL. CAV2-ΔE3-CGS was not detected in experimental animal farm. CAV was detected in dog detention center, and the positive rates were 5.88% (5/85) in oral swabs, 8.24% (7/85) in anal swabs, and 4% (1/25) in environmental samples. Conclusion The dual fluorescent quantitative PCR method can be used for the detection of CAV2-ΔE3-CGS after immunization and the investigation of CAV infection. The present study has shown that no CAV2-ΔE3-CGS has been detected after immunization and CAV infection rate of stay dogs is low in Shanghai. CAV2-ΔE3-CGS oral immunization meets requirement and is applicable.

PURPOSE: Details of patients hospitalized for asthma exacerbation in mainland China are lacking. To improve disease control and reduce economic burden, a large sample survey among this patient population is indispensable. This study aimed to investigate the clinical characteristics and outcomes of such patients.METHODS: A retrospective study was conducted on patients hospitalized for asthma exacerbation in 29 hospitals of 29 regions in mainland China during the period 2013 to 2014. Demographic features, pre-admission conditions, exacerbation details, and outcomes were summarized. Risk factors for exacerbation severity were analyzed.RESULTS: There were 3,240 asthmatic patients included in this study (57.7% females, 42.3% males). Only 28.0% used daily controller medications; 1,287 (39.7%) patients were not currently on inhaled corticosteroids. Acute upper airway infection was the most common trigger of exacerbation (42.3%). Patients with severe to life-threatening exacerbation tended to have a longer disease course, a smoking history, and had comorbidities such as hypertension, chronic obstructive pulmonary disease (COPD), and food allergy. The multivariate analysis showed that smoking history, comorbidities of hypertension, COPD, and food allergy were independent risk factors for more severe exacerbation. The number of patients hospitalized for asthma exacerbation varied with seasons, peaking in March and September. Eight patients died during the study period (mortality 0.25%).CONCLUSIONS: Despite enhanced education on asthma self-management in China during recent years, few patients were using daily controller medications before the onset of their exacerbation, indicating that more educational efforts and considerations are needed. The findings of this study may improve our understanding of hospital admission for asthma exacerbation in mainland China and provide evidence for decision-making.
Adrenal Cortex Hormones ; Asthma ; China ; Comorbidity ; Disease Progression ; Education ; Female ; Food Hypersensitivity ; Hospitalization ; Humans ; Hypertension ; Inpatients ; Medication Adherence ; Mortality ; Multivariate Analysis ; Pulmonary Disease, Chronic Obstructive ; Retrospective Studies ; Risk Factors ; Seasons ; Self Care ; Smoke ; Smoking

Objective To investigate the constitution and drug resistance status of the major pathogens in mechanically ventilated patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD),so as to provide evidences for clinical treatment.Methods From patients with AECOPD undergoing mechanical ventilation in EICU or ICU of the Second Affiliated Hospital of Wenzhou Medical University from January 2016 to December 2017,various specimens were collected for identification and drug susceptibility testing of pathogens;the clinical data and test results were analyzed.Results A total of 104 patients were included in the study.Total of 163 strains of pathogens were isolated from the specimens,of which gram-negative pathogens were the most common,accounting for 74.8％ of any pathogens.Drug resistance analysis showed that gram-negative pathogens were severely resistant to third generation cephalosporins,and were more sensitive to Sulperazon and carbapenems;gram-positive cocci were more sensitive to vancomycin and linezolid.Fungi were more sensitive to amphotericin B and 5-fluorocytosine.Most of the pathogens were multi-resistant.Conclusions In selecting antibiotics for the treatment of critical patients with AECOPD,emphasis should be placed on bacterial culture and drug susceptibility testing,so as to reduce irrational drug use and the emergence of drug-resistant strains or superinfection.

Objective To explore the role of receptor for advanced glycation end products (RAGE) in HMGBl-mediated CD4+ T cells differentiation to Th1/Th2.Methods CD4+ T lymphocytes isolated from the spleens of male BALB/C mice by magnetic beads were suspended in RPMI-1640 with 10％ FCS in 2× 106cell/well on 96-well cell culture plates in vitro.The cells were randomly divided 4 groups according to concentration of HMGB1 treatment:control group,10 ng/mL group,100 ng/mL group,1 000 ng/mL group after stimulation with ConA in 3 μg/mL for 12 hours.IL-4 and IFN-γ levels in culture supernatants were quantitated by ELISA kits after HMGB1 stimulation for 12,24,and 48 h.According to the results,cultured cells were exposed to HMGB1 in 100 ng/mL for 24 h in the following experiments.The cells were randomly divided into 4 groups:control group,A group,B group,C group,and each group were cultured with ConA in 3 μg/mL for 24 h.The cells of control group and other three groups were stimulated with PBS or 100 μg/L HMGB1 for 24 h.The cells of B,C groups were incubated with 1/200 diluted 5 μg/L anti-RAGE Abs (anti-bodies) or PBS for 2 h before HMGB1 stimulation.The cell suspension was obtained to detect the levels of IL-4 and IFN-γ by EILSA and the protein levels and mRNA expressions of RAGE,CATA-3 were detected by western-blot and real-time fluorescent quantitative PCR,respectively.ResuRs Compared with control group,CD4+ T cells incubated with increasing concentrations of HMGB1 (100,1 000 ng/mL) for 24 h resulted in a decrease in IFN-γ/IL-4 ratio (P＜0.05).When CD4+ T cells were exposed to 100 ng/mL HMGB1 for 12 h,IFN-γ/IL-4 ratio was markedly increased.However,CD4+ T cells treated with 100 ng/mL HMGB1 for 24,48 h,IFN-γ/IL-4 ratio was markedly inhibited (P＜0.05).Compared with control group,protein levels and mRNA expressions of RAGE and GATA3 of cells in A group were significantly increased (P＜0.05),and IFN-γ/IL-4 ratio of cell suspension in A group and B group was significantly down-regulated (P＜0.05).Compared with A group,IFN-γ/IL-4 ratio of cell suspension in C group was increased (P＜0.05),and expression of GATA3 mRNA was down-regulated (P＜0.05).Compared with A group,protein level of RAGE of cells in C group was significantly down-regulated (P＜0.05),but protein level of RAGE of cells in C group was still increased compared with control group (P＜0.05).Conclusion Th1/Th2 differentiation induced by HMGB1 on CD4+ T lymphocytes in vitro was at least partly mediated by over-activating RAGE/GATA3 pathway.

Objective To investigate the effect of continuous renal replacement therapy ( CRRT) on the reg-ulation of Treg/Th17 in patients with severe sepsis,and related inflammatory factors IL-6,IL-17,IL-10,and TNF-α.Methods 60 patients with severe sepsis were randomly divided into two groups,30 cases in each group.The control group received conventional treatment,and the observation group was treated with CRRT on the basis of the control group.Flow cytometry was used to detect the levels of Tregs and Th17 cells,and IL-6,IL-10,IL-17 and TNF-αwere detected by ELISA method.At the same time,the APACHEII score,ICU length of hospital stay were observed and recorded.Results After treatment,APACHE Ⅱscore,ICU length of hospital of the observation group were lower than the control group,there were statistically significant differences(t=4.258,t=4.518,all P<0.05). The expression levels of Th17,Treg and Tregs/Th17 in the observation group were significantly lower than the control group,the differences were statistically significant (t=5.872,4.267,4.285,all P<0.05).The expression levels of IL-6,IL-17,TNF-αin the observation group decreased more significantly than the control group,the differences were statistically significant (t=5.829,5.257,5.983,all P<0.05).But the expression level of IL-10 in the two groups had no obvious change[(35.26 ±18.63) pg/mL vs (35.52 ±20.09) pg/mL,t=2.164,P>0.05].Conclusion CRRT can not only remove the inflammatory mediators of abnormal expression,improve the function of T cells,but also can maintain the balance between Th17 and Treg,improve the immune disorders,and improve the prognosis of sepsis.

Objective To evaluate of impact of thalidomide on CD4 +CD25 +T lymphocytes in patients with acute myeloid leukemia and its clinical curative effect.Methods 71 cases of patients with AML were randomly divided into the control group and the experiment group, and 31 healthy people were selected to be the normal group.The experiment group and the control group patients were treated with the same chemotherapy, the experiment group was treated with thalidomide.The levels of CD4 +CD25 +T lymphocyte, CD3 +T lymphocyte, CD4 +T lymphocyte, CD8 +T lymphocyte, ratio of CD4 +/CD8 +and NK cells were detected at before treatment, 10 to 14d after treatment, complete remission 6 months follow-up.In normal group, the same index was detected before and after chemotherapy, and 10-14 d.The clinical curative effect of the experiment group and the control group were observed.Results The effective rate of the experiment group was higher than of the control group(P<0.05); The levels of CD4 +CD25 +T lymphocyte in the experiment group and control group were significantly higher than in the normal group(P<0.05), the two groups decreased compared with the control group, the level of CD4 +CD25 +T lymphocyte in the experiment group was significantly decreased(P<0.05).And with thalidomide treatment, the experiment group in the CD3 +T lymphocytes, CD4 +T lymphocytes and ratio of CD4 +/CD8 +, NK cells were significantly higher than the control group ( P<0.05 ) .Conclusion Thalidomide can improve both the immunity cell function and the clinical efficiency in patients with AML.The mechanism is related to reduce the level of CD4 +CD25 +T lymphocyte.

Objective To explore the viral etiology spectrum of different age and different seasons for neo-natal pneumonia.Methods Medical records of 1 073 cases of neonatal pneumonia in our hospital were retrospectively analyzed,line direct immunofluorescence assay were used to detect nasopharyngeal secretions of newborns,the test results were statistically analyzed.Results In 1 073 cases with neonatal pneumonia,406 cases were detected positive with virus infected,the positive rate was 37.8%,7 cases were mix infected.334 cases were infected by respiratory syncytial virus(RSV),which had the highest detection rate,accounting for 82.3%;RSV infection rate in 1 -6 month baby was 36.4%,which was higher than the >6 -12 month -old baby with RSV infection rate 26.5%,the difference was statistically significant(χ2 =12.25,P <0.05);RSV infection rate in winter and spring group was 39.7%,which was significantly higher than that in autumn and winter group(13.6%);PIV3 infection rate in winter and spring group was 1.8%,significantly lower than that in autumn and summer group(9.1%),the difference was statistically signifi-cant(χ2 =31.27,P <0.05 ).Conclusion RSV is the most common viral in neonatal pneumonia,more attention should be payed to RSV infection control in small babies and at winter and spring,pay attention to PIV3 infection at autumn and summer.