Objective To investigate the phenotype and the immunoregulatory function of CD8αα+TCRαβ+regulatory T cells in peripheral blood samples from mice.Methods The distribution profile and the phenotype of CD8αα+TCRαβ+regulatory T cells in C57BL/6 mice were detected by flow cytometry.The cytokines released by CD8αα+TCRαβ+regulatory T cells upon the stimulation with anti-CD3 antibody were analyzed by cytometric bead array.The in vitro immunosuppressive activity of CD8αα+TCRαβ+regulatory T cells on activated CD4+T cells was analyzed by using flow cytometry and carboxyfluorescein succinimidyl ester ( CFSE ) .An adoptive cell transfer assay was set up to evaluate the immunoprotective effects of CD8αα+TCRαβ+ regulatory T cells in a mouse model of experimental autoimmune encephalomyelitis ( EAE) .Results CD8αα+TCRαβ+regulatory T cells were detected in liver, spleen and peripheral blood samples collected from na?ve C57BL/6 mice.Compared with CD8αβ+TCRαβ+regulatory T cells, CD8αα+TCRαβ+regulatory T cells showed a memory-activated phenotype of CD25+CD122high CD44high CD62Llow CD69high NK1.1+DX5+.CD8αα+TCRαβ+regulatory T cells could produce IL-2 after 24 hours stimulation with anti-CD3 antibody, followed by producing IFN-γ, TNF-α, IL-4, IL-17A and traces of IL-6 and IL-10. In vitro, CD8αα+TCRαβ+regulatory T cells specifically suppressed the proliferation of activated CD4+T cells ( P<0.01 ).Moreover, they could delay the onset of EAE in mice and reduce clinical score (P<0.01).Conclusion CD8αα+TCRαβ+regulatory T cells were a unique population with immunoregula-tory function, which could be used as a potential therapeutic target in the treatment of autoimmune disease.

Objective To study the changes of trans-diaphragmatic pressure (Ptra) and its correlation with esophageal pressure (Peso) through ARDS piglet model.Methods Five piglets were enrolled in the study.Peso,gastric pressure (Pgas) and intra-thoracic pressure (Pint) was monitored through balloon inserted.The data before ARDS serve as control.ARDS was produced in the piglets through saline lavage.The pressure were observed and the Ptra were calculated.The pressure changes and correlation between Ptra and Peso were analyzed as well.Linear regression with the coefficient of determination and t-test were used as appropriate.Significance was assumed for P ＜ 0.05.Results Peso,Pgas and Pint before ARDS were 7.3 ± 1.9,25.5 ± 2.4,- 1.23 ± 0.21 cmH2O,Ptra was 18.2 ± 1.6 cmH2O.While after ARDS,the data were 4.7 ± 1.4,31.1 ± 3.1 and - 1.79 ± 0.28 cmH2O,and Ptra was 26.4 ± 2.1 cmH2 O,and all these changes were obviously ( P ＜ 0.05 ).The correlation between Pint and Peso,Pint and Ptra (A) and Ptra ( B ) were 0.93 ± 0.025,0.88 ± 0.023 and 0.87 ± 0.37 before ARDS.After ARDS,the correlation changed to be 0.82 ±0.21,0.81 ±0.20 and 0.78 ±0.31.Although a bit decreased,the correlation was still positive (P ＜ 0.01 ).Conclusions There existed good correlations between Peso and Ptra as well as between Pint and Peso before or after ARDS.Ptra was increased obviously after ARDS,which could lead to respiratory muscle fatigue.

OBJECTIVE To explore the correlation between CYP2C9*2, CYP2C9*3, CYP4F2, and VKORC1 1173C>T polymorphisms and warfarin maintenance dose, and establish an artificial neural network prediction model for international normalized ratio(INR) values after warfarin administration to improve the accuracy of stable dose prediction. METHODS A retrospective study was conducted by collecting clinical data and warfarin pharmacogenetic data from 214 warfarin-treated patients who achieved a stable anticoagulant state from 2019 to 2021. The impact of clinical factors and various gene phenotypes on the patient's warfarin steady-state dose was analyzed. A machine learning prediction model was established by simulating the input of the patient's warfarin dose to calculate the INR target and predict the steady-state dose. The accuracy of the model was compared with the direct dose prediction method and the multiple regression model. RESULTS The multiple regression model had the highest accuracy rate of 56.4% for predicting the patient's steady state dose in the dataset. The machine learning prediction model had a mean absolute error(MAE) of 0.40 and R2 of 0.81 when inputting the steady state dose to predict the INR value. Directly predicting the dose resulted in a MAE of 0.52 and R2 of 0.68. After group training, the error rate decreased by 20.4% and the accuracy increased by 7.3%. CONCLUSION The artificial neural network model for predicting INR using simulated input of warfarin dose can more accurately predict patient's steady-state dose, which facilitates individualized dosing and promotes the development of precision medicine.

OBJECTIVETo investigate the distribution of Warfarin related genes and the relationship between genotype, gender, weight, age and the administrative dose of Warfarin in Shanghai area.

METHODSThe clinical data (including sex, age and administrative dose of Warfarin) of 214 patients with stable warfarin dose and the international normalized ratio (INR) between 1.5-3.0 were collected. Polymerase chain reaction-high resolution melting (PCR-HRM) technique was used to detect the single nucleotide polymorphisms (SNPs) of CYP2C9*2 rs1799853, CYP2C9*3 rs1057910, CYP4F2 rs2108622 and VKORC1 rs9934438. The associations of genotype data with clinical material, including gender, age, weight and warfarin dosage were analyzed.

RESULTSAmong 214 patients, 99.53% (213 cases) patients with CC (wild type) of CYP2C9*2 rs1799853 and only 1 case with CT (heterozygous mutation) ; 92.52% (198 cases) with AA (wild type), 7.48% (16 cases) with CA (heterozygous mutation) of CYP2C9*3rs1057910; about 57.94% (124 cases) with CC(wild type) of CYP4F2 rs2108622, the CT and TT (heterozygous and homozygotic mutation) accounted for 42.06% (90 cases). In SNP VKORC1 rs9934438, 82.71% (177cases) were TT (wild type), 17.29% (37 cases) CT (heterozygous mutation). There are no significant difference (P=0.0872) in patients with maintenance dose in CYP2C9*3 between AA and CA gene mutations[(2.816±1.055) mg/d vs (2.352±0.805)mg/d], and no significant difference (P=0.5954) of that in CYP4F2 between CC and CT+TT gene mutations [(2.736±1.062) mg/d vs (2.813±1.034) mg/d]; but the significant differences (P=0.0001) does exist in patients with maintenance dose in VKORC1 between TT and CT variants [(2.597±0.866) mg/d vs (3.660±1.350) mg/d]. The warfarin maintain dosage was negatively correlated with the average age (r=-0.9669) and positively correlated with the body weight (r=0.9022).

CONCLUSIONIt is of great significance to detect the VKORC1 variants for warfarin dosage adjustment in Shanghai population. However, the detection of CYP2C9*2 and CYP4F2 polymorphisms had no significant associations for warfarin dosage adjustment.

Adult ; Aged ; Aged, 80 and over ; Anticoagulants ; administration & dosage ; Body Weight ; China ; epidemiology ; Cytochrome P-450 CYP2C9 ; genetics ; Cytochrome P-450 Enzyme System ; genetics ; Cytochrome P450 Family 4 ; Dose-Response Relationship, Drug ; Female ; Genotype ; Heterozygote ; Humans ; International Normalized Ratio ; Male ; Middle Aged ; Sex Distribution ; Vitamin K Epoxide Reductases ; genetics ; Warfarin ; administration & dosage ; Young Adult

Objective:To explore the value of shear wave elastography imaging(SWE) in the diagnosis of renal allograft fibrosis and analyze its advantages and limitations.Methods:The renal allograft of 61 patients who underwent renal allograft biopsy from June 2019 to April 2020 in the First Affiliated Hospital of Sun Yat-sen University were included in this study. According to the Banff classification, there were 51 patients with mild-degree fibration(interstitial fibrosis/tubular atrophy, IFTA 0-Ⅰ), and 29 patients with moderate or severe-degree fibration(IFTA Ⅱ-Ⅲ). Two-dimensional ultrasound, color Doppler flow imaging, SWE and kidney function test performed. All the results were compared between the two groups. Diagnostic performance was evaluated by receiver operating characteristic (ROC) curve using pathology as gold standard. The sensitivity, specificity, positive predictive value and negative predictive value were calculated for the diagnosis of moderate or severe-degree fibration.Results:Compared to mild-degree fibration group, creatinine( P<0.001), glomerular filtration rate( P<0.001), RI of arcuate arteries( P=0.022) and SWE value( P<0.001) significantly increased in the moderate or severe-degree fibration group. There were significant correlations between IFTA and creatinine ( r s=0.488, P<0.001), glomerular filtration rate ( r s=-0.452, P<0.001), RI of arcuate arteries( r s=0.228, P=0.042), SWE value( r s=0.584, P<0.001). Taking the cutoff value of SWE value deduced by ROC curve as 21.7 kPa, the area under ROC curve was 0.827. The sensitivity, specificity, positive predictive value and negative predictive value were 86.2%, 74.5%, 61.0% and 89.7%, respectively. Conclusions:There is a good correlation between the SWE value and the degree of fibrosis in the transplanted kidney. SWE can be used to distinguish mild from moderate or severe fibrosis of renal allograft, providing a potential noninvasive method for the assessment of kidney allograft fibration.