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

Background::Findings on the association of genetic factors and colorectal cancer (CRC) survival are limited and inconsistent, and revealing the mechanism underlying their prognostic roles is of great importance. This study aimed to explore the relationship between functional genetic variations and the prognosis of CRC and further reveal the possible mechanism.Methods::We first systematically performed expression quantitative trait locus (eQTL) analysis using The Cancer Genome Atlas (TCGA) dataset. Then, the Kaplan-Meier analysis was used to filter out the survival-related eQTL target genes of CRC patients in two public datasets (TCGA and GSE39582 dataset from the Gene Expression Omnibus database). The seven most potentially functional eQTL single nucleotide polymorphisms (SNPs) associated with six survival-related eQTL target genes were genotyped in 907 Chinese CRC patients with clinical prognosis data. The regulatory mechanism of the survival-related SNP was further confirmed by functional experiments.Results::The rs71630754 regulating the expression of endoplasmic reticulum aminopeptidase 1 ( ERAP1) was significantly associated with the prognosis of CRC (additive model, hazard ratio [HR]: 1.43, 95% confidence interval [CI]: 1.08-1.88, P = 0.012). The results of dual-luciferase reporter assay and electrophoretic mobility shift assay showed that the A allele of the rs71630754 could increase the binding of transcription factor 3 (TCF3) and subsequently reduce the expression of ERAP1. The results of bioinformatic analysis showed that lower expression of ERAP1 could affect the tumor immune microenvironment and was significantly associated with severe survival outcomes. Conclusion::The rs71630754 could influence the prognosis of CRC patients by regulating the expression of the immune-related gene ERAP1. Trial Registration::No. NCT00454519 (https://clinicaltrials.gov/)

Drastic surges in intracellular reactive oxygen species (ROS) induce cell apoptosis, while most chemotherapy drugs lead to the accumulation of ROS. Here, we constructed an organic compound, arsenical N-‍(4-(1,3,2-dithiarsinan-2-yl)phenyl)acrylamide (AAZ2), which could prompt the ROS to trigger mitochondrial-dependent apoptosis in gastric cancer (GC). Mechanistically, by targeting pyruvate dehydrogenase kinase 1 (PDK1), AAZ2 caused metabolism alteration and the imbalance of redox homeostasis, followed by the inhibition of phosphoinositide-3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway and leading to the activation of B-cell lymphoma 2 (Bcl2)/Bcl2-associated X (Bax)/caspase-9 (Cas9)/Cas3 cascades. Importantly, our in vivo data demonstrated that AAZ2 could inhibit the growth of GC xenograft. Overall, our data suggested that AAZ2 could contribute to metabolic abnormalities, leading to mitochondrial-dependent apoptosis by targeting PDK1 in GC.
Humans ; Signal Transduction ; Stomach Neoplasms/drug therapy* ; Reactive Oxygen Species/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Apoptosis ; Proto-Oncogene Proteins c-bcl-2 ; Cell Line, Tumor

OBJECTIVE To establish and apply a method for simultaneous determination of the contents of dicentrine， protopine and coptisine in Tibetan Corydalis pallida of different origins， and to provide reference for origin determination and quality control of the kind of medicinal materials. METHODS Ultra performance liquid chromatography-tandem quadrupole mass spectrometry method was used. The determination was performed on Agilent EC-C18 column （100 mm×2.1 mm， 2.7 μm） with mobile phase consisted of acetonitrile-0.1% formic acid by gradient elution. The flow rate was 0.2 mL/min， and the column temperature was set at 35 ℃ . MS detection was carried out by electrospray ionization in positive modes， multiple reaction monitoring mode was used for quantitative analysis. RESULTS The injection mass concentrations of dicentrine， protopine， coptisine ranged from 5.88 to 117.60， 53.70 to 1 074.00， and 4.85 to 97.00 ng/mL， respectively， showing a good linear relationship with their respective peak areas （r＝0.998 2， 0.991 9， and 0.999 6， respectively）. The limits of quantitation were 2.35， 1.07 and 1.46 ng/mL； the limits of detection were 1.17， 0.54， 0.49 ng/mL， respectively. RSDs of precision， stability （24 h） and repeatability tests were all lower than 2.0％. The average recovery rates were 97.41%， 98.89% and 105.44%（ all RSDs＜5.0％， n＝6）. CONCLUSIONS The established method has good selectivity and high accuracy， and is suitable for the rapid analysis of dicentrine， protopine and coptisine in Corydalis. The total contents of three alkaloids in different original medicinal materials are from high to low in order of C. chrysosphaera， C. mucronifera， C. pygmaea， C. hendersonii and C. conspersa. The alkaloid contents in C. chrysosphaera and C. mucronifera are relatively similar， but no dicentrine has been detected in C. conspersa and C. hendersonii.

BACKGROUND: Immune checkpoint inhibitors (ICIs) are increasingly used as first-line therapy for patients with advanced non-small cell lung cancer (NSCLC) harboring no actionable mutations; however, data on their efficacy among patients presenting with intracranial lesions are limited. This study aimed to explore the efficacy and safety of ICIs combined with chemotherapy in advanced NSCLC patients with measurable brain metastasis at initial diagnosis. METHODS: Our study retrospectively analyzed clinical data of a total of 211 patients diagnosed with driver gene mutation-negative advanced NSCLC with measurable, asymptomatic brain metastasis at baseline from Hunan Cancer Hospital between January 1, 2019 and September 30, 2021. The patients were stratified into two groups according to the first-line treatment regimen received: ICI combined with chemotherapy ( n = 102) or chemotherapy ( n = 109). Systemic and intracranial objective response rates (ORRs) and progression-free survival (PFS) were analyzed. Adverse events were also compared between the groups. RESULTS: Compared with the chemotherapy-based regimen, the ICI-containing regimen was associated with a significantly higher intracranial (44.1% [45/102] vs . 28.4% [31/109], χ2 = 5.620, P = 0.013) and systemic (49.0% [50/102] vs . 33.9% [37/109], χ2 = 4.942, P = 0.019) ORRs and longer intracranial (11.0 months vs . 7.0 months, P <0.001) and systemic (9.0 months vs . 5.0 months, P <0.001) PFS. Multivariable analysis consistently revealed an independent association between receiving ICI plus platinum-based chemotherapy as a first-line regimen and prolonged intracranial PFS (hazard ratio [HR] = 0.52, 95% confidence interval [CI]: 0.37-0.73, P <0.001) and systemic PFS (HR = 0.48, 95% CI: 0.35-0.66, P <0.001). No unexpected serious adverse effects were observed. CONCLUSION: Our study provides real-world clinical evidence that ICI combined with chemotherapy is a promising first-line treatment option for driver gene mutation-negative advanced NSCLC patients who present with brain metastasis at initial diagnosis. CLINICAL TRIAL REGISTRATION https://www.clinicaltrials.gov/ , OMESIA, NCT05129202.
Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Immune Checkpoint Inhibitors/therapeutic use* ; Retrospective Studies ; Brain Neoplasms/genetics*

Objective:To investigate the effect of automated nested multiplex PCR system in the detection of children with severe community-acquired pneumonia(CAP), and identify the pathogenic infection of the children with severe CAP in Foshan.Methods:Children with severe CAP, who were admitted to the PICU at Foshan First People′s Hospital from January 2016 to December 2018, were enrolled in the analysis.Nasopharyngeal secretions were collected.And automated nested multiplex PCR was used to detect adenovirus, coronavirus (HKUl type, NL63 type, 229E type, 0C43 type), human metapneumovirus, influenza A virus (H1 subtype, H1-2009 subtype, H3 subtype), influenza B virus, parainfluenza virus (type 1, type 2, type 3, type 4), respiratory syncytial virus, Bacillus pertussis, Chlamydia pneumoniae and Mycoplasma pneumonia.Results:Among the 290 specimens detected by the automated nested multiplex PCR, 246(84.83%) were positive.There were 166 positive samples for a single pathogen, 60 positive samples for two pathogens, 17 positive samples for three pathogens, and three positive samples for four pathogens.Among the virus-positive cases, respiratory syncytial virus was the most common pathogen in children younger than 6 months(62.39%, 73/117). The most common pathogen was human rhinovirus/enterovirus(43.48%, 20/46) from seven months to one year old.Adenovirus(37.50%, 18/48) was the most common pathogen among children aged one to three years old.Rhinovirus/enterovirus(35.00%, 7/20) was the most common pathogen among children aged three to six years old.The most common pathogen in children over six years old was influenza virus(46.67%, 7/15). The adenovirus detection rate was highest in May, the syncytial virus detection rate was highest in August, and the influenza virus detection rate was highest in July.Mycoplasma pneumoniae and pertussis were distributed throughout the year.Conclusion:The automated nested multiplex PCR system can detect multiple pathogens efficiently, quickly and accurately; the common pathogens of severe CAP are diverse in different age groups; the epidemic season of common pathogens is unique in different regions due to different climates.

Objective To investigate the distribution characteristics and changing tendency of urinary tract stones.Methods From January 2011 to May 2017,clinical data of 15 269 patients treated in our center was retrospectively reviewed.The stone components were detected by the automatic stone infrared spectroscopy system and the predominant components were recorded.There were 9 019 male patients and 6 250 female patients.The patients were divided into four groups according to their age,including group A ≤ 18 years;group B 19-40 years;group C 41-60 years;and group D ＞ 60 years.Compared the distribution characteristics of urinary tract stones of patient in different groups of sex,age and calendar year.Results Calcium oxalate stones were more prevalent in males than females [6 221 (69.0％)vs.3 582 (57.3％),P ＜ 0.001],but calcium phosphate stones [210 (3.4％) vs.210 (2.3％)],magnesium ammonium phosphate stones [230(3.7％) vs.165 (1.8％)] and carbonate apatite stones [1 328 (21.3％) vs.1 030 (11.4％)] were more common in females than males (P ＜ 0.001,respectively).The proportion of uric acid stones in group D [679(20.7％)] was higher than that in group A [23(9.1％)],group B[260(7.9％)],group C [1 163 (13.8％)] (P ＜0.001,respectively).The peak of carbonate apatite stones was showed in group B [652(19.7％)] (P＜0.001,respectively).Ammonium urate stones [9(3.5％)] and cystine stones [36 (14.2％)] were more frequent in group A(P ＜0.001,respectively).In adults,the percentage of uric acid stones increased with age,such as group B [260(7.9％)],group C [1 163(13.8％)],group D [679 (20.7％)].And the carbonated apatite stones decreased with age,such as group B [652 (19.7％)],group C [1 270(15.1％)],group D [416(12.7％)] (P ＜0.001,respectively).Further analysis showed the proportion of calc ium oxalate (OR =0.944,95 ％ CI 0.927-0.962,P ＜ 0.001),ammonium urate stones (OR =0.854,95％ CI 0.742-0.982,P =0.027) decreased,while calcium phosphate (OR =1.192,95％ CI 1.127-1.261,P ＜0.001),uric acid (OR =1.042,95％ CI 1.015-1.069,P =0.002) and ammonium magnesium phosphate (OR =1.078,95％ CI 1.019-1.141,P =0.009) stones increased with time.Conclusions The distribution of stones was different in genders and age.Calcium oxalate stones were more common in male patients,while ammonium magnesium phosphate and carbonate apatite stones were more common in female patients.Uric acid stones were more frequent in patients older than 60,while carbonate apatite were more frequent in the 19-40 age group.The proportion of calcium oxalate and ammonium urate stones showed a downward trend,whereas calcium phosphate,uric acid and magnesium ammonium phosphate stones increased with time.

Objective: To observe the relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia and evaluate the safety preliminarily.Methods: The relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia were studied by the cough model caused by the irritation of ammonia water and the phenol red output of trachea in mice.The acute toxicity test and maximum tolerance test were carried out to evaluate the safety.Results: The total alkaloid in Atalantia buxifolia at low dose could obviously prolong cough incubation period and decrease cough times in mice, and that at high dose could significantly increase the secretion of phenol red in respiratory tract, and compared with those in the blank group, the differences were statistically significant (P<0.05).In the acute toxicity test, no death showed after the administration with maximum tolerance dosage, and the rate of weight growth had no difference between the blank group and the model group (P>0.05).Conclusion: The relieving cough and reducing sputum effects of total alkaloid in Atalantia buxifolia are notable in the cough model caused by the irritation of ammonia water and the phenol red output of trachea in mice.The maximum tolerable dose test shows the total alkaloid in Atalantia buxifolia is safe.

Objective To investigate SCCmec types in clinical isolates of methicillin-resistant Staphylococcus epidermidis (MRSE) carrying psm-mec.Methods We collected 165 strains of Staphylococcus epidermidis identified by automated microbiological identification system and screened MRSE by PCR amplification of esp and mecA gene.Strains with psm-mec were identified by amplification of psm-mec,fudoh and p221 DNA fragment;mec,ccr and SCCmec typing was conducted by multiplex PCR assay.Results Among 138 strains of MRSE,29 strains were identified as MRSE with psm-mec,and the carrying rate was 17.58%.Results of mec and ccr typing by multiple PCR showed that MRSE with psm-mec carried Class A mec,but the ccr type had obvious diversity.Results of SCCmec typing showed that all strains with psm-mec belonged to type Ⅱ and/or Ⅲ SCCmec.Conclusion Clinical isolates of MRSE with psm-mec carry homologous type Ⅱ and/or Ⅲ SCCmec harboring Class A mec.