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

ObjectiveTo explore the effects of Naozhenning granules on the memory function and neuron cells in the rat model of post-concussion syndrome based on mitochondrial biosynthesis. MethodSPF-grade Wistar rats were used to establish the multiple cerebral concussion （MCC） model by the weight-drop method. The successfully modeled rats were assigned into model， piracetam （0.324 g·kg^-1）， and low-， medium-， and high-dose （2.25， 4.5， and 9 g·kg^-1， respectively） Naozhenning groups. The rats were administrated with corresponding drugs by gavage and those in the blank group and model group were administrated the same volume of normal saline once a day for 14 days. The general state of rats was observed before and after treatment. The open field test and new object recognition test were conducted to examine the motor and memory abilities of rats. Hematoxylin-eosin staining was employed to observe the pathological changes of cortical neurons in rats. Western blot and real-time polymerase chain reaction were employed to determine the protein and mRNA levels， respectively， of peroxisome proliferator-activated receptor γ-coactivator-1α （PGC-1α）， nuclear respiratory factor-1 （NRF-1）， and transcription factor A mitochondrial （TFAM） in rat cortex. ResultCompared with the blank group， the model group showed anxious and manic mental status， yellow and messy fur， and reduced food intake. In the open field experiment， the model group showed reduced total movement distance， times of entering the central grid， and times of rearing decreased and increased resting time compared with the blank group （P<0.01）. The model group had lower recognition index of new objects than the blank group （P<0.01）. In addition， the modeling caused reduced neurons with sparse distribution and deformed， broken， and irregular nucleoli and down-regulated the mRNA and protein levels of PGC-1α， NRF-1， and TFAM in the cortex （P<0.01）. Compared with the model group， piracetam and Naozhenning improved the mental state， coat color， food intake， and activities of rats. In the open field test， piracetam and Naozhenning increased the total movement distance， the times of entering the central grid， and the times of rearing and shortened the resting time （P<0.05， P<0.01）. The piracetam and Naozhenning groups had higher recognition index of new objects than the model group （P<0.05， P<0.01）. Compared with the model group， the piracetam and Naozhenning groups showed increased neurons with tight arrangement and large and round nuclei， and some cells with irregular morphology and turbid cytoplasm. Furthermore， piracetam and medium-dose Naozhenning upregulated the protein levels of PGC-1α， NRF-1， and TFAM （P<0.01）. Low-dose Naozhenning upregulated the protein levels of NRF-1 and TFAM （P<0.01）， and high-dose Naozhenning upregulated the protein levels of PGC-1α and TFAM in the cortex （P<0.01）. The mRNA levels of PGC-1α， NRF-1， and TFAM in the cortex were upregulated in the piracetam group and Naozhenning groups （P<0.05， P<0.01）. ConclusionNaozhenning granules can improve the motor， memory， and learning， repair the neuronal damage， and protect the nerve function in the rat model of MCC by promoting mitochondrial biosynthesis.

Objective To investigate the role of squalene epoxidase(SQLE)knockout in anti-tumor effect vial im-proving CD8+T cell infiltration in melanoma tumor microenvironment.Methods Both immunodeficient and immu-nocompetent mice were inoculated with SQLE knockout B16F10 cells to determine the cell-autonomous and non-au-tonomous regulation of malignancy.Antibody blockade,Luminex multiplex assays,and flow cytometry were em-ployed to explore the impact of SQLE gene knockout on the secretion of cytokines/chemokines and immune cell in-filtration.Bioinformatics analysis was conducted to validate the correlation between SQLE expression and immune infiltration as well as clinical prognosis in melanoma patients.Results Compared with immunodeficient mice,SQLE knockout significantly inhibited melanoma proliferation in immunocompetent mice and prolonged their surviv-al.SQLE knockout induced the secretion of cytokines and chemokines from tumor cells,improved CD8+T cell in-filtration in the tumor microenvironment,thereby potentiating anti-tumor immunity.Bioinformatics analysis sugges-ted a significant correlation between SQLE and its corresponding immune infiltration markers with the prognosis of melanoma patients.Conclusion SQLE regulates anti-tumor immunity by controlling cytokines and chemokines re-leasing in tumor microenvironment,thus holding promise as a novel tumor immunotherapy target and efficacy predic-tion molecular indicator.

OBJECTIVETo investigate the protective effect of bone marrow mesenchymal stem cells (BMSCs)-derived exosomesagainst testicular ischemia-reperfusion injury (IRI) in rats.

METHODSRat BMSCs were isolated, cultured and identified in theprimary culture. The exosomes were extracted from the BMSCs and characterized using nanoparticle tracking analysis, transmission electron microscopy, and Western blotting. Twenty-four healthy male SD rats were randomly divided into shamoperation group, testicular IRI with saline treatment group and IRI with exosome treatment group. The contralateral testes ofthe rats were collected for pathological observation, aseessment of superoxide dismutase (SOD) and malondialdehyde (MDA), and detection of HMGB1, caspases-3 and cleaved caspase-3 expressions using Western blotting.

RESULTSWe successfullyobtained exosomes from rat BMSCs. Testicular IRI significantly impaired testicular spermatogenesis, which was markedlyimproved by treatment with the exosomes ( < 0.05). Testicular IRI also caused significant increase in the protein expression ofHMGB1, caspase-3 and cleaved caspase-3 in the testicular tissue, and treatment with the exosomes obviously amelioratedthese changes ( < 0.05).

CONCLUSIONSBMSCs-derived exosomes protects against testicular IRI due to the anti-oxidant, antiinflammatory and anti-apoptosis activities of the exosomes.

Objective To investigate the role of vitamin D in the synthesis and degradation of aggrecan in rat articular chondrocytes at cellular level. Methods Rat articular chondrocytes were stimulated by IL-1α, IL-1β and TNF-α, respectively. Normal and inflammatory chondrocytes were treated with different doses of vitamin D, respectively. CCK8, Flow cytometry, real time-PCR and western blot analysis were used to examine the proliferation activity and apoptosis level of chondrocytes, and the expression of aggrecan, ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels. Results IL-1α,IL-1β and TNF-α significantly decreased the proliferation activity and increased the apoptosis level of the chondrocytes. Furthermore, IL-1α, IL-1β and TNF-α significantly decreased the expression of aggrecan, and increased the expressions of ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels in the chondrocytes. 1α,25 (OH)2D3supplementation significantly increased the proliferation activity and decreased the apoptosis level of chondrocytes stimulated by IL-1α, IL-1β and TNF-α in a dose-dependent manner, but not affected the normal chondrocytes. Meanwhile, 1α,25(OH)2D3also significantly increased the expression of aggrecan, and decreased the expressions of ADAMTS-4 and ADAMTS-5 at both mRNA and protein levels in the chondrocytes under inflammatory conditions. Conclusions Vitamin D may promote the anabolism of aggrecan and inhibit aggrecanase activity in chondrocytes under inflammatory conditions, which may impact overall protection for articular cartilage.

Objective To explore the doctor-nurse integrated health care management model in perioperative nursing management of transvaginal natural orifice transluminal endoscopic surgery (NOTES). Methods A total of 70 female patients underwent transvaginal NOTES. All patients were randomly divided into the control and the study groups. The control group adopted the traditional model of care, and the study group implemented a layered panel system of integrated health care nursing mode, which consisted of three lines (leader), second line, front-line doctors, anesthesiologists consisting of the doctor series and secondary responsibility leader, a responsibility of nurses, operating room nurses laparoscopy group traveling together constituted a series of nurses′ health care integration of the health care team. The average length of stay, preoperative waiting days, awareness of the disease, early postoperative activities and reasonable diet, patient satisfaction with care, doctors′ satisfaction with nurses′ operation and overall satisfaction with health care work were compared. Results The preoperative waiting days, average length of stay, surgical satisfaction, patients′ satisfaction, doctors-nurses′overall satisfaction in the study group were (3.95±0.51) d, (11.58±1.83) d, (95.48±3.82)%, (99.26± 0.61)%, (96.33±0.29)%, the control group were (5.50±0.68) d, (15.79±2.14) d, (82.08±7.21)%, (92.18± 3.54)%, (80.03 ± 5.88)%, the differences were statistically significant (t=-4.431-6.172, P < 0.05). The patients awareness of the disease, early postoperative activities and reasonable diet (postoperative eating time) in the study group were (92.77 ± 0.59)%, (1.60 ± 0.63) d, (2.20 ± 0.54) d, the control group were (93.15 ± 0.47)%, (1.90 ± 0.42) d, (2.40 ± 0.33) d, the difference was not statistically significant (t=-0.898,-0.771,-0.201, P>0.05). Conclusions The implementation ofdoctor-nurse integrated health carenew model can optimize the collocation of medical health care integration, enhance the quality of care, improve patient experience, and improve patients′and doctors′satisfaction for nursing work.

OBJEVTIVETo investigate the expression of cysteine-rich secretory protein 2 (CRISP2) in spermatozoa of patients with asthenozoospermia and explore its clinical significance.

METHOSSemen samples were collected from 24 normal volunteers and 24 patients with asthenozoospermia for detecting CRISP2 mRNA and protein expressions using qRT-PCR and Western blotting, respectively. The correlation of CRISP2 expressions with sperm morphology, progressive motility and fertility prognosis were analyzed in patients with asthenozoospermia.

RESULTSCRISP2 protein expression was obviously lowered in the ejaculated spermatozoa of patients with asthenozoospermia as compared to the normal volunteers, but no significant difference in CRISP2 mRNA expression was found between the two groups. Correlation analysis showed that CRISP2 protein expression was positively correlated with normal sperm morphology (r=0.6182, P=0.0037) and progressive motility (r=0.6309, P=0.0029). Follow-up study of the patients revealed a higher fertility rate in patients with a relatively high CRISP2 protein expression than in those with low CRISP2 protein expression (80.0% vs 20.0%, P=0.0230).

CONCLUSIONThe expression level of CRISP2 protein is positively correlated with normal sperm morphology and progressive motility. A reduced CRISP2 protein expression indicates poor fertility prognosis of patients with asthenozoospermia, suggesting the potential value of CRISP2 as a novel therapeutic target for treating asthenozoospermia.

Asthenozoospermia ; metabolism ; Case-Control Studies ; Fertility ; Follow-Up Studies ; Glycoproteins ; metabolism ; Humans ; Male ; RNA, Messenger ; Sperm Motility ; Spermatozoa ; metabolism

OBJECTIVETo evaluate the effect of placement of urethral catheter combined with lidocaine on urethral irritation caused by postoperative indwelling catheters.

METHODSA total of 120 male surgical patients requiring postoperative indwelling catheters between June 2011 and January 2012 were divided into two equal groups for placement of painless urethral catheter combined with bladder washing with lidocaine on the first postoperative day, or for routine catheter placement only. The symptoms of urethral irritation such as urethral pain, urinary urgency, and perineal discomforts were observed and compared between the two groups.

RESULTSIn patients with painless urethral catheter placement combined with bladder washing with lidocaine, 11 developed urethral irritation symptoms, as compared to 24 in the patients with routine catheter placement only, showing a significant difference between the two groups (P<0.05).

CONCLUSIONPlacement of painless urethral catheter combined with bladder washing with lidocaine can significantly reduce the incidence of urethral irritation due to postoperative indwelling catheters.

Adult ; Humans ; Lidocaine ; Male ; Middle Aged ; Urinary Catheterization ; methods ; Urinary Catheters ; adverse effects ; Urination Disorders ; etiology ; prevention & control ; Young Adult

OBJEVTIVETo analyze the complications of extracorporeal shock wave lithotripsy (ESWL) for treatment of lower urinary tract stones and identify the factors contributing to the complications.

METHODSWe retrospectively analyzed the postoperative complications of ESWL in 83 patients with lower urinary tract stones and their association with the patients' age, height, body weight, body mass index (BMI), stone features, shock wave times, and shock wave energy.

RESULTSOf the 83 cases treated with ESWL, 72.3% (60 cases) showed gross hematuria, 27.7% (23 cases) complained of pain, and 25.3% (21 cases) had both pain and gross hematuria. None of the patients reported vomiting, skin rash, flatulence, gastrointestinal bleeding or other complications. Patients with postoperative gross hematuria, pain, and both had significantly greater stone length, stone width and more shock body times than those without these complications (P<0.01 or P<0.05). Patients with gross hematuria had significantly lower height than those without complications (P<0.05).

CONCLUSIONThe common complications of ESWL for lower urinary tract stones include hematuria and pain. The patient's height, stone length, stone width and shock times are important factors contributing to these postoperative complications.

Abdominal Pain ; etiology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Hematuria ; etiology ; Humans ; Lithotripsy ; adverse effects ; Male ; Middle Aged ; Retrospective Studies ; Urinary Bladder Calculi ; therapy ; Young Adult

High mobility group A2 protein (HMGA2), an architectural factor, is highly expressed in various cancer types including lung cancers. It is a candidate target for cancer therapy. RNAi is an effective gene silencing method with low cost and less time-consuming. It is possible to exploit this technology in therapy. Here, 5 siRNAs targeting Hmga2 gene (HMGA2 siRNA1-5) were designed and synthesized. MTT assay, colony formation assay, transwell assay and flow cytometry were used to evaluate the effects of these siRNAs on lung cancer cell lines (NCI-H446 and A549). Results from cell proliferation, clone formation, migration and apoptosis showed that HMGA2 siRNA1, 3, 5 could affect these aspects for both lung cancer cell lines. Among the five siRNAs, HMGA2 siRNA5 showed the greatest inhibition effects. The inhibition effects of HMGA2 siRNA5 are sequence specific and are not due to the induction of interferon response. Taken together, siRNAs targeting Hmga2 gene are potential candidates for lung cancer gene therapy.