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

Objective:To investigate the predictive value of heparin binding protein(HBP) combined with pediatric sequential organ failure assessment(pSOFA) in children with sepsis.Methods:Children with sepsis admitted to PICU of Hunan Provincial People's Hospital (the First Affiliated Hospital of Hunan Normal University) from January 2021 to June 2022 were selected as study group,while those who underwent elective surgery for inguinal hernia and assessment of precocious puberty and short stature during the same period were selected as control group.All children with sepsis were divided into sepsis group and septic shock group according to their severity as well as survival group and death group according to prognosis.The study group was monitored for HBP on the 1st,3rd,and 7th day of admission,while the control group was monitored for HBP on the 1st day of admission.Patients in the sepsis group received pSOFA scores immediately after admission.The laboratory results and HBP concentrations were compared between groups，and a joint model was established in combination with pSOFA to observe its predictive performance in sepsis prognosis.Results:A total of 50 children with sepsis were included in study group，including 45 children with sepsis and five children with septic shock.There were 27 males and 23 females，aged 1 month～13 years（median age two years）.There were 7 deaths in this study，including two patients with sepsis and five patients with septic shock.The HBP concentration in the study group was significantly higher than that in the control group on the 1st day，and the HBP concentration in the group gradually decreased with the prolongation of hospital stay.The concentration of HBP on the first day of septic shock group was higher than that of sepsis group，and the difference was statistically significant（ P<0.001）.The concentration of HBP on the 1st day in the sepsis death group was significantly higher than that in the sepsis survival group（ P=0.023）.The receivor operator characteristic curve analysis showed that HBP and pSOFA had good predictive value for the death of children with sepsis，and the joint model of HBP and pSOFA（75.1×pSOFA-0.1×HBP）had the best predictive performance for the death of children with sepsis，but there was no significant difference with the pSOFA. Conclusion:The HBP level significantly increases in children with sepsis，and gradually decreases with the length of hospital stay，and HBP has great value in predicting the outcome of death in children with sepsis，and the combination of pSOFA could improve its predictive ability of death,but not better than pSOFA.

Community-acquired pneumonia(CAP) is one of the leading causes of death in children under 5 years of age.Early identification and clarification of its severity and appropriate therapeutic measures can improve survival, but there are limitations in the existing laboratory indices applied to diagnose CAP.Therefore, it is still necessary to find new and highly specific biomarkers that can identify the etiology and predict the severity of the disease before it worsens in children, and provides a basis for more effective therapeutic measures.Metabolomics provides a new way to search for biomarkers and pathogenesis through qualitative and quantitative analysis of metabolite changes in biological samples.This review summarized the latest research progress on metabolomics in childhood CAP, hoping to provide ideas for the early diagnosis and treatment of childhood CAP.

The heterogeneity of exhausted T cells (Tex) is a critical determinant of immune checkpoint blockade therapy efficacy. However, few studies have explored exhausted T cell subpopulations in human cancers. In the present study, we examined samples from two cohorts of 175 patients with head and neck squamous cell cancer (HNSCC) by multiplex immunohistochemistry (mIHC) to investigate two subsets of Tex, CD8⁺PD1⁺TCF1⁺ progenitor exhausted T cells (TCF1⁺Tex^prog) and CD8⁺PD1⁺TCF1^- terminally exhausted T cells (TCF1^-Tex^term). Moreover, fresh tumor samples from 34 patients with HNSCC were examined by flow cytometry and immunohistochemistry to further investigate their properties and cytotoxic capabilities and their correlation with regulatory T cells (Tregs) in the tumor immune microenvironment (TIME). mIHC and flow cytometry analysis showed that TCF1^-Tex^term represented a greater proportion of CD8⁺PD1⁺Tex than TCF1⁺Tex^prog in most patients. TCF1⁺Tex^prog produced abundant TNFα, while TCF1^-Tex^term expressed higher levels of CD103, TIM-3, CTLA-4, and TIGIT. TCF1^-Tex^term exhibited a polyfunctional TNFα⁺GZMB⁺IFNγ⁺ phenotype; and were associated with better overall survival and recurrence-free survival. The results also indicated that larger proportions of TCF1^-Tex^term were accompanied by an increase in the proportion of Tregs. Therefore, it was concluded that TCF1^-Tex^term was the major CD8⁺PD1⁺Tex subset in the HNSCC TIME and that these cells favor patient survival. A high proportion of TCF1^-Tex^term was associated with greater Treg abundance.
CD8-Positive T-Lymphocytes ; Head and Neck Neoplasms/therapy* ; Humans ; Immunotherapy/methods* ; Prognosis ; Programmed Cell Death 1 Receptor ; Squamous Cell Carcinoma of Head and Neck/therapy* ; Tumor Microenvironment ; Tumor Necrosis Factor-alpha

Objective To explore the clinical value of Z score in assessing coronary artery lesions (CAL) of children with Kawasaki disease. Methods The clinical records of 102 children with Kawasaki disease from January 2012 to December 2016 in Gansu Provincial Hospital were retrospectively analyzed. The internal diameter of left main coronary artery (LMCA) and right coronary artery ( RCA) was measured by echocardiography (ECHO),and the incidence of CAL was preliminarily judged. The Z scores of LMCA and RCA were calculated on the basis of the coronary artery diameter,the age of the children and the body surface area,and the incidence of CAL was judged again. Results A total of 22 cases(21. 6%) of CAL were found in 102 cases by ECHO examination,of which 18 cases(17. 6%) of LMCA lesions,and 22 cases(21. 6%) of RCA lesions. A total of 33 cases(32. 4%) of CAL were found by calculating the Z score of coronary artery, of which 29 cases(28. 4%) of LMCA lesions and 33 cases(32. 4%) of RCA lesions. There was significant difference between two methods for determining LMCA lesions (χ2=3. 35,P＜0. 05),and there was no sig-nificant difference between two methods for determining RCA lesions (χ2=3. 01,P＞0. 05). Z score of coro-nary artery was more accurate to detect the CAL in Kawasaki disease,especially LMCA lesions. A large coro-nary artery aneurysm was found in the patients with the largest Z score by selective coronary angiography. Conclusion The Z score can be more conductive to assess the CAL in children with Kawasaki disease,and the higher the Z score,the more serious the CAL is.

Objective To analyze genetic testing and prenatal diagnosis of two pedigrees with X-linked ichthyosis.Methods Karyotyping,bacterial artificial chromosomes-on-BeadsTM (BoBs),fluorescence in situ hybridization (FISH) and single nucleotide polymorphism array (SNP-array) were used to detect amniotic fluid and peripheral blood specimens of two pedigrees,one with and one without known family history of ichthyosis.Clinical data was collected and analyzed as well.Results (1) The pedigree without known family history:Prenatal BoBs showed that the XC1 probe of fetus Ⅳ-12 was from 0.36 to 0.50,suggesting the presence of microdeletion.SNP-array analysis of gravida Ⅲ-13 showed a 1.68 Mb copy number deletion at Xp22.31 and four missing Online Mendelian Inheritance in Man (OMIM) genes (HDHD1,STS,VCX and PNPLA4).Fetal SNP-array revealed a deletion of arr[hg19] Xp22.31 (6 455 151-8 135 644)× 0,indicating a maternally inherited one.FISH analysis verified the deletion in STS gene in fetus Ⅳ-12,whose karyotype was 46,XY.The gravida's female cousin (Ⅲ-21) and nephew (Ⅳ-14) also had STS gene deletion,which size was the same as that from the gravida and the fetus.Fetus (Ⅳ-12) was delivered at term by cesarean section with normal skin,but an extensive white scales appeared on the abdomen one week after birth and the symptom was aggravated when the weather was dry.The infant was followed up to eight months old and no other clinical symptoms were found.(2) The pedigree with known family history:SNP-array revealed that a 1.2 Mb copy number deletion at Xp22.31 and four missing OMIM genes (HDHD1,STS,VCX and PNPLA4) were detected in pregnant women (Ⅲ-21),proband (Ⅳ-16) and fetus (Ⅳ-17).FISH analysis of the fetus verified the deletion in STS gene.The karyotype of the fetus was 46,XY.Fetus Ⅳ-17 was delivered at term by cesarean section with normal skin,but white scales widely appeared on the abdomen ten days after birth.The infant was followed up to four months old and no other clinical symptoms were found.Conclusion Molecular genetic techniques such as BoBs,FISH and SNP array are used in combination in this study to provide genetic testing and prenatal diagnosis to two XLI pedigrees,which is helpful for clinical diagnosis and genetic counseling.

Objective To investigate the value of ultrasound screening for congenital cytomegalovirus (CMV) infection in fetuses and to summarize the clinical manifestations.Methods From January 2012 to December 2017,we retrospectively analyzed the clinical data of 905 gravidas who received invasive prenatal diagnosis in Fujian Provincial Maternity and Children's Hospital for abnormal prenatal ultrasound findings including ventriculomegaly,intracranial calcification,microcephaly,echogenic bowel and fetal growth restriction (FGR).CMV DNA loads in amniotic fluid and neonatal urine were detected by real-time polymerase chain reaction.CMV-specific IgM and IgG in umbilical cord and neonatal peripheral blood were detected by commercial enzyme 1 inked immunosorbent assay kits.Eighteen fetuses with normal karyotype were diagnosed as congenital CMV infection.Relationships of ultrasound features and CMV DNA loads in amniotic fluid to pregnancy outcomes were analyzed with x2 test or Fisher's exact test.Results (1) Congenital CMV infection was detected in 18 fetuses in this study with an detection rate of 1.99％ (18/905).Three pregnancies were terminated immediately after the diagnosis was confirmed,two terminated when the ventriculomegaly progressed,five terminated for hydrocephaly and eight continued to delivery.(2) Congenital CMV infection rate was significantly higher in those with two or more ultrasound abnormalities than that in those with only one abnormal indicator [3.92％(8/204) vs 1.28％(9/701),x2=4.619,P=0.032].Fetuses with craniocerebral abnormalities were more likely to have congenital CMV infection than those without [3.11％(13/418) vs 0.82％(4/487),x2=6.392,P=0.012].(3) Among the 18 fetuses with congenital CMV infection,those with serious ultrasound abnormalities had a significantly higher rate of adverse outcomes than those without (11/11 vs 3/7,Fisher's exact test,bilateral P=0.043).No significant difference in the rate of adverse outcomes was found between fetuses with low and high CMV DNA loads in amniotic fluid (3/4 vs 12/14,Fisher's exact test,bilateral P=1.000).Conclusions Ultrasound abnormalities including ventriculomegaly,intracranial calcification,microcephaly,echogenic bowel and FGR,especially those with multiple abnormalities and brain abnormalities,increased risk of congenital CMV infection.Congenital CMV fetuses with serious ultrasound abnormalities has adverse outcomes.

[Objective]To investigate the effect of Donepezil on dysgnosia in patients with mild-moderate general paralysis of the insane(GPI).[Methods]Forty-seven patients with GPI were assigned into study group(n=24)or control group(n=23). All these patients were treated with a large dose of penicillin for 2 weeks and then with Benzathine for 3 weeks. And the Donepezil admin-istered to patients in study group.The mini-mental state examination(MMSE)and the clinical dementia rating(CDR)were scored as the measurement of efficiency.[Result]The scores of the MMSE and CDR were not statistically different between the two groups at baseline and 4 weeks after treatment,and both were increased 12 weeks after treatment(P<0.05),especially in study group as com-pared with those in control group(P < 0.05).[Conclusion]Donepezil as an adjunctive therapy may be effective for dysgnosia in patients with mild-moderate GPI. Donepezil is also safe and well tolerated.

Background: Aberrant Bcl-2 transcription is closely related with nodal diffuse large B-cell lymphoma (DLBCL), however, the relationship between Bcl-2 and primary gastrointestinal DLBCL (PGI-DLBCL) was not fully studied.Aims: To investigate the relationship between Bcl-2 gene amplification and protein expression and clinicopathological characteristics, immunophenotype and prognosis of PGI-DLBCL.Methods: Clinical data was collected from 136 PGI-DLBCL patients receiving surgical treatment, and a telephone interview was conducted for survival information.Bcl-2 gene amplification and protein expression in tumor tissue were determined by fluorescence in situ hybridization and immuno-histochemistry, respectively, and relationships between Bcl-2 and clinicopathological characteristics, immunophenotype and prognosis of PGI-DLBCL were analyzed.Results: Among 136 PGI-DLBCL patients, 33 (24.3%) showing gene amplification and 90 (66.2%) showing protein expression of Bcl-2;gene amplification was correlated with primary tumor location, Ann Arbor stage, serum lactate dehydrogenase level, B symptom and International Prognostic Index (IPI) score (P<0.05), while protein expression was correlated with primary tumor location and immunophenotype (P<0.05).5-year overall survival (OS) in patients positive for Bcl-2 gene amplification and patients with non-GCB immunophenotype and positive for Bcl-2 protein expression were inferior to those negative ones (41.5%vs.71.5%, P<0.05;54.6% vs.84.6%, P<0.05).In Bcl-2 gene amplification or protein expression positive patients, 5-year OS of CHOP chemotherapy was inferior to that of rituximab combined with CHOP chemotherapy (48.6%vs.80.3%, P<0.05;66.4%vs.83.4%, P<0.05).Conclusions: Detection of Bcl-2 gene amplification is useful for prediction of prognosis in PGI-DLBCL.Both patients with Bcl-2 gene amplification and non-GCB patients with Bcl-2 protein expression have a poorer prognosis.Rituximab may improve the prognosis in patients with Bcl-2 gene amplification or protein expression.

Objective To explore the effect on efficacy and safety of Wuling capsule on patients with post-stroke depression.Methods From July 2012 to September 2014, 90 post-stroke depression patients were selected in our hospital.The patients were divided into treatment group (n=45) and control group (n=45) by random number table method.The post-stroke depression patients in control group were treated with fluoxetine as clinical drug program.The post-stroke depression patients in treatment group were treated with Wuling capsule as clinical drug program, six weeks for a course of treatment, both group were treated after two courses,the clinical indicators improve effect, clinical treatment, changes of cytokines, incidence of adverse reactions of the patients between two groups were compared and analyzed.Results Hamilton depression scale (HAMD) scores and Barther index (BI) scores of patients in treatment group post-treatment were better than control group (P<0.05) .Compared with control group, the ratio of clinical efficiency and total efficiency significant in treatment group were increased significantly (P<0.05).The patients in treatment group post-treatment, the levels of cytokine of interleukin-6 (IL-6) and tumor necrosis factor-α(TNF-α) decreased significantly (P<0.05) .Compared with control group, the incidences of adverse effect of patients in treatment group were decreased (P<0.05).Conclusions Wuling capsule has certain therapeutic advantages for improving clinical efficacy and reducing adverse reactions for patients with post-stroke depression.It could as the therapy of choice for post-stroke depression patients.