OBJECTIVE To investigate the clinical efficacy of pramipexole combined with levodopa-benserazide in the treatment of Parkinson’s disease （PD）. METHODS A total of 108 PD patients treated at the Fifth People’s Hospital of Wuhu City from January 1， 2021， to February 28， 2023， were randomly divided into observation group and control group， with 54 cases in each group. Patients in the control group were administered levodopa-benserazide （initial dose of 62.5 mg per dose）， three times daily； after one month， the dose was increased to 250 mg per dose， four times daily. Patients in the observation group received the same treatment as the control group， with the addition of pramipexole （initial dose of 0.25 mg per dose） orally twice daily on an empty stomach； after 14 days， the dose was increased to 0.25 mg per dose， three times daily. Both groups were treated for 3 months. The short-term efficacy， safety and long-term prognosis of the two groups were compared. RESULTS After treatment， the observation group had significantly lower scores on the Unified Parkinson’s Disease Rating Scale part Ⅲ （UPDRS-Ⅲ）， the Hamilton Depression Scale （HAMD）， the Hamilton Anxiety Scale （HAMA）， and the Parkinson’s Disease Questionnaire-39（ PDQ- 39） compared to the control group； in contrast， the observation group had higher scores on the Montreal Cognitive Assessment （MoCA） scale， the Mini-mental State Examination （MMSE） scale， the Mattis Dementia Rating Scale （DRS）， and the Quality of Life （QOL） scale （P＜0.05）. Both groups showed a significant reduction in UPDRS-Ⅲ and PDQ-39 scores， and a significant increase in DRS scores compared to baseline （P＜0.05）. However， only the observation group showed a significant increase in MoCA scale， MMSE scale， and QOL scores， and a significant decrease in HAMD and HAMA scores compared to baseline （P＜ 0.05）. The total incidence of adverse drug reactions in both groups was not significantly different （P＞0.05）. The 12 months follow-up results showed that the incidence of dementia and mortality rates in the observation group were significantly lower than the control group （P＜0.05）. CONCLUSIONS Pramipexole combined with levodopa-benserazide significantly improves motor function， cognitive function， quality of life and symptoms of depression and anxiety in PD patients， and may reduce the long-term risk of dementia and mortality in these patients.

Objective:To explore the clinical significance of hepatitis B virus (HBV) DNA detection in screening patients with hepatitis B.Methods:Clinical data of 682 331 hepatitis B patients were retrospectively analyzed. The HBV DNA of these patients was detected in the Fifth Medical Center of the PLA General Hospital from January 2017 to December 2021, there were 481 159 males and 201 172 females in this cohort, the average age was (41.34±16.13) years. Patients were divided into HBV DNA positive group (219 879 cases) and HBV DNA negative group (462 452 cases). Clinical characteristics, data of five serologic markers of hepatitis B and hepatitis B surface antigen quantification (HBsAg-QN), liver function, alpha fetoprotein (AFP) and prothrombin time (PT) results were collected and analyzed and compared between the two groups.Results:The positive rate of HBV DNA was 32.22% (219 879/682 331) in this cohort. Among the different age groups, the positive rate of HBV DNA was the highest (40.34%, 128 038/317 380) in young people aged 18-44 years. The proportion of patients was lower among aged <1, 45-59 and ≥60 years patients in HBV DNA positive group than that in HBV DNA negative group, while the proportion of patients was higher among aged 1-17 and 18-44 years patients in HBV DNA positive group than that in HBV DNA negative group (all P<0.001). Among 2 291 <1-year-old infants tested for HBV DNA, 71 infants were HBV DNA positive. The positive rates of HBV DNA from 2017 to 2021 were 4.86% (27/556), 3.68% (14/380), 3.47% (17/490), 1.55% (6/386) and 1.46% (7/479) respectively, showing a downward trend year by year. The positive rate of HBV DNA in acute hepatitis B (AHB) patients was the highest (49.88%, 208/417) among 680 040 patients with hepatitis B. The proportion of AHB patients (0.09%, 208/219 808) and chronic hepatitis B (80.44%, 176 806/219 808) in HBV DNA positive group was higher than that in HBV DNA negative group [0.05% (209/460 232) and 65.45% (301, 216/460 232)], while the proportion of patients with HBV-related liver cirrhosis (11.28%, 24 793/219 808), HBV-related liver cancer (6.72%, 14 775/219 808), liver cancer surgery (1.39%, 3 055/219 808) and liver transplantation (0.08%, 171/219 808) were lower than that in HBV DNA negative group [22.99% (105 813/460 232), 7.25% (33 385/460 232), 3.50% (16 129/460 232) and 0.76% (3 480/460 232)] (all P<0.001). At the same time, positive rate of hepatitis B surface antigen (HbsAg), HBsAg-QN, hepatitis B e antigen (HbeAg), level of total bilirubin, total bilirubin, AFP and PT were higher in HBV DNA positive group than those in HBV DNA negative group, while the age, male ratio and albumin results in HBV DNA positive group were lower than those in HBV DNA negative group (all P<0.01). The HBV DNA loads were higher in HBsAg positive group, hepatitis B surface antibody positive group and HBeAg positive group than those in respective negative groups, while the HBV DNA loads were lower in hepatitis B e antibody positive group and hepatitis B core antibody positive group than those in respective negative groups (all P<0.001). Conclusions:The mother to child transmission rate of<1-year-old infants decreases year by year. HBV DNA is an important factor for the progression of hepatitis B disease. HBV DNA positive hepatitis B patients with higher HBsAg-QN values are more likely to have abnormal serum markers such as liver dysfunction. HBV DNA detection is therefore of clinical importance in screening patients with hepatitis B.

Objective:To investigate the changes and significance of granulocyte-like myeloid-derived suppressor cells (G-MDSC) in the acute phage of Kawasaki disease (KD).Methods:Forty-two children with acute KD were enrolled in the present study and 32 age-matched healthy children were selected as control group. The proportion of HLA-DR -CD11b + CD33 + CD14 -CD15 + G-MDSC, the concentration of reactive oxygen species (ROS) and the expression of arginase-1 (Arg-1), programmed death-ligand 1 (PD-L1), cytotoxic T lymphocyte associated protein 4 (CTLA4), glycoprotein 130 (gp130) and phosphorylated signal transducer and activator of transcription 3 (pSTAT3) at protein level were detected by flow cytometry. Quantitative real-time PCR was used to measure the expression of inducible nitric oxide synthase (iNOS), interferon regulatory factor 8 (IRF-8), IL-6 receptor α subunit (IL-6Rα), granulocyte colony-stimulating factor receptor (G-CSFR), CCAAT/enhancer binding protein β (C/EBPβ), suppressor of cytokine signaling 1 (SOCS1) and SOCS3 at mRNA level in G-MDSC. Chromatin immunoprecipitation was performed to detect the acetylation of histone H3 at the promoters of SOCS1 and SOCS3 genes. Plasma concentrations of IL-6 and granulocyte colony-stimulating factor (G-CSF) and protein levels of IL-10, transforming growth factor-β (TGF-β) and nitric oxide (NO) in the culture supernatant of G-MDSC stimulated with LPS were measured by ELISA. Results:(1) Compared with the control group, the proportion of HLA-DR -CD11b + CD33 + CD14 -CD15 + G-MDSC as well as the concentration of ROS and the expression of inhibitory molecules (Arg-1, PD-L1 and CTLA4) in G-MDSC increased significantly in patients with acute KD ( P<0.05). Moreover, the concentrations of IL-10 and TGF-β in culture supernatant of G-MDSC were also higher than those of the control group after stimulation with lipopolysaccharide for 48 h ( P<0.05). All of the seven afore-mentioned indexes in KD patients with coronary artery lesion (CAL group ) were lower than those in patients without coronary artery lesion (NCAL group) ( P<0.05), and restored to some extent after IVIG therapy ( P<0.05). There were no statistical differences in iNOS expression or NO concentration in culture supernatant of G-MDSC among different groups ( P<0.05). (2) Plasma concentrations of IL-6 and G-CSF, and the expression of IL-6Rα, gp130, G-CSFR, pSTAT3 and C/EBPβ increased remarkably during acute phase of KD ( P<0.05). The expression of IRF-8 at transcription level in patients with acute KD was found to be lower than that of healthy controls ( P<0.05), and restored significantly after IVIG therapy ( P<0.05). Moreover, the plasma concentrations of IL-6 and G-CSF and the expression of IL-6Rα, gp130, G-CSFR and IRF-8 in the CAL group were higher than those in the NCAL group ( P<0.05), while the expression of pSTAT3 and C/EBPβ was lower in the CAL group ( P<0.05), which were restored by IVIG therapy ( P<0.05). (3) In patients with acute KD, the expression of SOCS1 and SOCS3 at mRNA level and histone acetylation at the promoters of SOCS1 and SOCS3 genes were reduced significantly in comparison with those in healthy controls ( P<0.05) , but were increased remarkably after IVIG treatment( P<0.05). The four indexes were higher in the CAL group than in the NCAL group ( P<0.05). Pearson correlation analysis showed the expression of SOCS1 and SOCS3 was negatively correlated with the protein level of pSTAT3 in G-MDSC of patients with acute KD ( r=-0.46 and -0.32, P<0.05). Conclusions:Changes in the number and function of G-MDSC caused by aberrant histone acetylation at SOCS1 and SOCS3 genes might contribute to the immune dysfunction and vascular damage in patients with KD.

Objective:To investigate the changes of CD8 + CD28 - regulatory T cells (Treg) and its role in the pathogenesis of Kawasaki disease (KD). Methods:A total of 48 children with KD were enrolled in the present study from June 2019 to December 2021. Blood samples were collected from them during acute phage of KD and after intravenous immunoglobulin (IVIG) treatment. Another 32 age-matched healthy children were recruited as control group. The proportions of CD8 + CD28 -Treg cells and the expression of programmed cell death protein 1 (PD-1), factor associated suicide ligand (FasL), inducible T-cell co-stimulator ligand (ICOSL), CD80 and CD86 protein were evaluated by flow cytometry. The expression of Helios, perforin, granzyme B, immunoglobulin-like transcript 3 (ILT3) and ILT4 at the transcription level was measured by real-time PCR. Concentrations of IL-10 and TGF-β in the culture supernatants of CD8 + CD28 -Treg cells stimulated with activated CD4 + T cells were measured by ELISA. Results:⑴ The proportions of CD8 + CD28 -Treg cells and the expression of Helios in patients with acute KD were higher than those in the control group ( P<0.05), and reduced remarkably after IVIG therapy ( P<0.05). The two afore-mentioned indexes were lower in patients combined with coronary artery lesion (CAL) than in those without coronary artery lesion (NCAL) ( P<0.05). ⑵ Compared with the control group, the patients with acute KD showed increased expression of FasL, PD-1, ICOSL and perforin in CD8 + CD28 -Treg cells ( P<0.05). The concentrations of IL-10 and TGF-β1 in the culture supernatants of CD8 + CD28 -Treg cells from patients with acute KD were lower than those in the control group after stimulation with activated CD4 + T cells ( P<0.05), which restored to some extent after IVIG treatment ( P<0.05). All of the six above-mentioned indexes in the CAL group were found to be lower than those in the NCAL group ( P<0.05). There were slight differences in granzyme B expression between different groups ( P>0.05). (3) In comparison with the healthy controls, the patients with acute KD showed overexpressed co-stimulatory molecules such as CD80 and CD86 on CD14 + cells ( P<0.05) and up-regulated expression of inhibitory molecules ILT3 and ILT4 ( P<0.05), which were restored remarkably after IVIG treatment ( P<0.05). Furthermore, the expression of CD80 and CD86 at protein level increased in the CAL group than in the NCAL group ( P<0.05), while the expression of ILT3 and ILT4 at transcriptional level decreased in the CAL group ( P<0.05). Conclusions:Relative insufficiency and impaired function of CD8 + CD28 -Treg cells might be one of the important factors resulting in immune dysfunction and vascular damage in KD patients.

Objective:To explore the effect of Notch1 signaling on regulatory T cells and its roles in vascular damage in patients with Kawasaki disease (KD).Methods:A total of 42 children with KD were enrolled in the present study from March 2019 to June 2020, as 32 age-matched healthy children were recruited as control. The proportions of CD4 +CD25 hiFoxp 3+ regulatory T cells (Treg) and expressions of transcription factor forkhead box protein 3 (Foxp3), cytotoxic T lymphocyte associated antigen-4 (CTLA4), glucocorticoid-induced tumor necrosis factor receptor (GITR), and Notch1 protein were evaluated by flow cytometry. Chromatin immunoprecipitation was conducted to detect acetylation level of histone H4 (H4Ac) associated with the promoter of Foxp3 gene and its binding abilities of Notch1 intracellular domain 1 (NICD1), recombination signal binding protein for immunoglobulin kappa J region (RBP-J) and p300 in CD4 + T cells. Transcription levels of Foxp3, presenilin 1 (PSEN1), mastermind like transcriptional coactivator 1 (MAML1), and RBP-J in CD4 + T cells were determined by real-time polymerase chain reaction (PCR). Concentrations of interleukin (IL)-10 and transforming growth factor-β (TGF-β) in plasma and culture supernatant stimulated with Jagged1 were measured by enzyme linked immunosorbent assay. Independent-sample t-test, Pearson correlation analysis was used as the statistical method in this study. Results:① The frequencies of Treg in acute KD patients decreased significantly [(4.3±1.5)% vs (7.9±2.9)%; t=6.41, P<0.001], as protein levels of Foxp3, CTLA4 and GITR and concentrations of IL-10 and TGF-β in plasma reduced remarkably in acute KD patients ( t=6.87, P<0.001; t=4.26, P<0.001; t=7.88, P<0.001; t=8.42, P<0.001; t=13.01, P<0.001). All parameters afore-mentioned in patients combined with coronary artery lesions (CAL) were lower than those of patients without coronary artery lesions (NCAL) ( t=5.83, P<0.001; t=3.83, P<0.001; t=3.28, P=0.002; t=5.05, P<0.001; t=5.96, P<0.001; t=5.17, P<0.001), and increased after therapy ( t=7.13, P<0.001; t=6.10, P<0.001; t=4.31, P<0.001; t=6.55, P<0.001; t=7.40, P<0.001; t=7.84, P<0.001). ② H4Ac associated with promoter of Foxp3 gene and the binding abilities of NICD1 and p300 in acute KD patients were lower than those of the controls ( t=10.25, P<0.001; t=6.93, P<0.001; t=6.75, P<0.001), and increased remarkably after therapy ( t=7.72, P<0.001; t=4.16, P<0.001; t=5.76, P<0.001). Meanwhile, the three items in CAL group were found to be less than those of NCAL group ( t=6.08, P<0.001; t=2.66, P=0.011; t=6.02, P<0.001). Pearson correlation analysis showed a positive correlation between H4Ac associated with Foxp3 promoter and its mRNA level in acute KD patients ( r=0.47, P<0.001). No statistical significant difference about the binding ability of RBP-J with Foxp3 promoter were found among the groups ( t=0.57, P>0.05; t=0.61, P>0.05; t=1.20, P>0.05). ③ Protein level of Notch1 and the expressions of PSEN1, MAML1 and RBP-J mRNA in CD4 + T cells from acute KD patients were down-regulated remarkably ( t=5.28, P<0.001; t=6.31, P<0.001; t=11.78, P<0.001; t=8.06, P<0.001), and restored after therapy ( t=4.77, P<0.001; t=6.43, P<0.001; t=11.95, P<0.001; t=7.79, P<0.001). In parallel, the four indexes aforementioned of CAL group were lower than those of NCAL group ( t=3.16, P=0.003; t=4.13, P<0.001; t=5.42, P<0.001; t=4.05, P<0.001). Upon rhJagged1 stimulation for 48 hours, H4Ac level of Foxp3 promoter and its binding abilities with NICD1 and p300 in CD4 + T cells in KD patients and control group was significantly higher than those of untreated group [(KD: t=15.36, P<0.001; t=7.25, P<0.001; t=14.29, P<0.001), (Ctrl: t=7.87, P<0.001; t=5.71, P<0.001; t=8.74, P<0.001)], as the binding ability of RBP-J with Foxp3 promoter increased slightly without statistically significant difference (KD: t=1.11, P>0.05; Ctrl: t=1.37, P>0.05). Simultaneously, H4Ac level of Foxp3 promoter and its binding abilities with NICD1 and p300 in KD group were still lower than those of the control group after stimulation ( t=3.86, P<0.001; t=3.42, P=0.001; t=2.85, P=0.006). ④ After incubation of PBMC from heathy children with KD serum, the proportion of Treg cells, protein level of Foxp3 and expressions of Notch1 and RBP-J in CD4 + T cells in the group treated with IVIG increased significantly compared with the untreated group ( t=7.10, P<0.001; t=10.16, P<0.001; t=8.06, P<0.001; t=9.77, P<0.001), as well as H4Ac level of Foxp3 promoter and its binding abilities with NICD1 in the group treat with IVIG were also higher than the latter ( t=7.24, P<0.001; t=8.24, P<0.001). Conclusion:Insufficiency and impaired function of Treg caused by aberrant Notch1 signaling may be the important factor contributing to immune dysfunction and vascular damage in KD.

Objective:In this article, we analyzed and discussed the clinical characteristics and S region gene sequencing of hepatitis B virus in HBsAg anti-HBs coexistent patients.Methods:Data of 5 serologic markers of hepatitis B and quantitative result, liver function and HBV DNA load of HBsAg positive patients were collected, and their basic clinical information were recorded. According to the positive and negative result of Anti-HBs, the clinical and virological characteristics of these two groups were analyzed. At the same time, among 17 320 patients with HBsAg positive HBV infection, 994 cases were tested by gene sequencing. The S region amino acid mutation, site mutation detection rate and genotype of 994 HBV infected patients with gene sequencing were statistically analyzed.Results:The positive rate of HBsAg and Anti-HBs was 4.36% (756/17 320). HBV-related cirrhosis in HBsAg+ /Anti-HBs+ group (19.71%) was significantly higher than that in HBsAg+ /Anti-HBs-group (15.94%), while chronic hepatitis B (62.04%) was significantly lower than that in HBsAg+ /Anti-HBs-group (67.06%). At the same time, the positive rates of HBsAg-quantification (QN) and ALT in HBsAg+ /Anti-HBs+ group were significantly lower than those in HBsAg+ /Anti-HBs-group, the positive rate of HBeAg was significantly higher than that in HBsAg+ /Anti-HBs-group, and the HBV DNA was higher than that in HBsAg+ /Anti-HBs-group, but the difference was no statistical significance. Gene sequencing was performed in 994 HBV patients. Genotype C (81.79%) had the highest proportion, genotype B (17.40%) was the second, and genotype D (0.80%) was the least in two groups. In genotype C HBV infected patients, the detection rate of sP120Q/T/A/S mutant in HBsAg+ /Anti-HBs+ group was significantly higher than that in HBsAg+ /Anti-HBs-group. Meanwhile, regardless of genotype B or C or overall comparison, the detection rate of sG145A/E/K/R mutant of HBV infected patients in HBsAg+ /Anti-HBs+ group was significantly higher than that in HBsAg+ /Anti-HBs-group, these differences were all statistically significant.Conclusions:The hepatitis B patients with coexistence of HBsAg and Anti-HBs were more likely to develop cirrhosis, and the hepatitis B patients with HBV gene sequencing results were mainly type C2. The drug resistance variation of S-region sP120Q/T/A/S and sG145A/E/K/R mutants of patients with HBV infection is an important reason for the coexistence of HBsAg and Anti-HBS.

Objective:To investigate the changes of monocytic myeloid-derived suppressor cells (M-MDSC) in children with acute Kawasaki disease (KD) and its roles in the immunological pathogenesis of KD.Methods:A total of 38 children with acute KD were enrolled in the present study and 32 age-matched healthy children were selected as control group. The proportions of HLA-DR -CD11b + CD33 + CD15 -CD14 + M-MDSC and CD4 + CD25 + CD127 - regulatory T cells (Treg) in peripheral blood, concentrations of reactive oxygen species (ROS) and expression of arginase-1 (Arg-1), CD39, CD73, CD40, CD40L and CCR5 at protein levels were detected by flow cytometry. Quantitative real-time PCR was used to evaluate the transcription levels of inducible nitric oxide synthase (iNOS) in M-MDSC and the transcription levels of cytotoxic T-lymphocyte associated antigen 4 (CTLA4) and lymphocyte-activation gene 3 (LAG3) in Treg. Concentrations of NO, CCL3, CCL4, CCL5, IL-10 and TGF-β in the supernatants of cell culture were measured by ELISA. Results:(1) The proportion of HLA-DR -CD11b + CD33 + CD15 -CD14 + M-MDSC, the concentration of intracellular ROS and the expression of iNOS, CD39 and CD73 in M-MDSC decreased significantly in patients with acute KD as compared with those in the control group ( P<0.05), and the concentrations of NO, IL-10 and TGF-β in culture supernatant of M-MDSC were lower than those in the control group upon lipopolysaccharide (LPS) stimulation for 48 h ( P<0.05). All of the aforementioned indexes restored to some extent after intravenous immunoglobulin (IVIG) therapy ( P<0.05). No statistical differences were found in Arg-1 expression between healthy controls and patients with KD before or after IVIG therapy ( P<0.05). (2) CD40 expression on M-MDSC was significantly lower in the acute KD group than in the control group ( P<0.05). The concentrations of CCL3, CCL4 and CCL5 in the culture supernatants of M-MDSC were lower in the acute KD group than in the control group after LPS stimulation ( P<0.05). With IVIG treatment, all of the indexes were up-regulated significantly ( P<0.05), although CD40 expression was still lower in the acute KD group than in the control group ( P<0.05). (3) The proportion of CD4 + CD25 + CD127 -Treg and the expression of CTLA4, LAG3, CD40L and CCR5 reduced significantly in patients with acute KD as compared those in healthy controls ( P<0.05), and all increased remarkably after IVIG therapy ( P<0.05). Pearson correlation analysis showed a positive correlation between the proportions of M-MDSC and Treg in patients with acute KD ( r=0.58, P<0.05). Conclusions:Insufficiency and impaired function of M-MDSC might be a major cause of immune dysfunction in patients with acute KD.

Objective:To investigate the histone acetylation of interleukin-4(IL-4) gene and its roles in immunological pathogenesis of Kawasaki disease (KD).Methods:Thirty-six children with KD and 28 age-matched healthy children in Shenzhen Children′s Hospital from October 2016 to December 2018 were recruited in this study.Peripheral venous blood samples were collected from healthy controls (28 cases) and patients with KD during acute phase and 4 to 5 days after effective intravenous immunoglobulin (IVIG) treatment.Co-immunoprecipitation followed by real-time PCR was used to assess histone H4 acetylation levels of IL-4 promoter and Va enhancer, and binding abilities of p300 and CREB-binding protein (CBP) with promoter and Va enhancer of IL-4 gene in peripheral blood CD4 + T cells.Flow cytometry was performed to analyze the proportion of CD4 + IL-4 + T cells (Th2) and protein le-vels of phosphorylated signal transducer and activator of transcription 6 (pSTAT6), GATA binding protein 3 (GATA3), nuclear factor 1 of activated T cells(NFAT1), transforming growth factor-β receptor Ⅱ (TGF-βRⅡ), and phosphorylated L-type amino acid transporter 1(pLAT1). Quantitative real-time PCR was used to evaluate the transcription levels of IL-4, IL-5, IL-13, IL-4 receptor α (IL-4Rα), transforming growth factor-β receptor Ⅰ (TGF-βRⅠ) and sex-determining region Y(SRY)-box 4 (SOX4) in CD4 + T cells.Plasma concentrations of IL-4 and transforming growth factor-β(TGF-β) were measured by enzyme-linked immunosorbent assay. Results:(1)Compared with control group, the proportion of Th2 cells, expression levels of Th2-associated cytokines (IL-4, IL-5 and IL-13) and histone H4 acetylation levels associating with IL-4 promoter and Va enhancer, increased remarkably during acute KD(all P<0.05), and restored after IVIG therapy(all P<0.05). Meanwhile, all the former items in KD patients with coronary artery lesions (CAL) were higher than those in patients with non-coronary artery lesions (NCAL) (all P<0.05). (2) Compared with control group, binding abilities of p300 and CBP with IL-4 promoter and Va enhancer in CD4 + T cells were up-regulated significantly during acute KD (all P<0.05), and decreased in varying degrees after IVIG treatment (all P<0.05). Positive correlations between binding abilities of p300 with IL-4 (promoter and Va enhancer) and the expression of IL-4 promoter and Va enhancer were detected in patients with acute KD ( r=0.72, 0.43, all P<0.05). Furthermore, binding abilities of p300 and CBP with IL-4 promoter and Va enhancer in CAL group were higher than those in NCAL group (all P<0.05). (3) Compared with control group, patients with acute KD had remarkably increased plasma concentration of IL-4, and expression levels of IL-4Rα/STAT6/GATA-3 and pLAT1/NFAT1 in CD4 + T cells (all P<0.05), and significantly down-regulated plasma concentration of TGF-β and expression level of TGF-βRⅡ/TGF-βRⅠ/SOX4 (all P<0.05). All the items mentioned above restored in varying degrees after IVIG treatment (all P<0.05). Simultaneously, the 6 items aforementioned in CAL group were found to be higher than those in NCAL group (all P<0.05), while the latter four items were lower than those in NCAL group (all P<0.05). Conclusion:Histone hyperacetylation of IL-4 gene may be related to immune dysfunction in patients with KD.

Objective To investigate the histone acetylation status of forkhead box P3(Foxp3)gene and its roles in immunological pathogenesis of Kawasaki disease(KD). Methods Forty - two children with KD and 32 age -matched healthy children consented to participate in this study as a control group. Co - immunoprecipitation and real -time PCR were performed to determine acetylation levels of histone H4 associated with Foxp3 gene and binding abilities of Kruppel like factor 10(KLF10)and p300 / CBP - associated factor (PCAF)with Foxp3 gene in CD4 + T cells. The proportion of CD4 + CD25 high Foxp3 + cells (Treg)and protein levels of Foxp3,KLF10,PCAF,phosphated SMAD family member 2 / 3(pSmad2 / 3)and itchy E3 ubiquitin protein ligase(Itch)were analyzed by flow cytometry. Quantitative real - time PCR was used to evaluate the levels of Foxp3,transforming growth factor - β1 receptor Ⅱ (TGF - βRⅡ), transforming growth factor - β1 receptor Ⅰ(TGF - βR Ⅰ),tyrosine kinase 2 (Tyk2),SH2 domain - containing protein - tyrosine phosphatase - 2(SHP2)and cytotoxic T - lymphocyte - associated protein 4 (CTLA4)mRNA in Treg. Plasma concentrations of TGF - β and interleukin - 6(IL - 6)were measured by enzyme - linked immunosorbent assay. Results (1)Compared with the control group,the proportion of Treg,expression levels of Foxp3 and TGF - β, and histone acetylation levels of Foxp3 promoter decreased remarkably during acute KD,and the differences were all statistically significant(all P < 0. 05),and restored after IVIG therapy(all P < 0. 05). Meanwhile,the 5 indexes afore-mentioned in KD patients with coronary artery lesions (KD - CAL +)were lower than those without coronary artery le-sions(KD - CAL -),and the differences were all statistically significant (all P < 0. 05). (2)Compared with the control group,protein levels of KLF10 and PCAF in Treg cells,and their binding abilities with Foxp3 gene were significantly down - regulated during acute KD,and the differences were all statistically significant(all P < 0. 05),and restored to some extent after IVIG treatment(all P < 0. 05). Positive correlations between protein levels of KLF10 and PCAF,and mRNA level of Foxp3 were detected in acute KD patients (r = 0. 47,0. 59,all P < 0. 05). Furthermore,protein levels of KLF10 and PCAF and their binding abilities with Foxp3 gene in KD - CAL + group were lower than those in KD -CAL - group,and the differences were all statistically significant(all P < 0. 05). (3)Compared with the control group, the plasma concentration of TGF - β and expressions of TGF - βRⅡ/ Ⅰ,pSmad2 / 3,Itch,CTLA4 and SHP2 in Treg were down - regulated during the acute phase of KD,and the differences were all statistically significant(all P < 0. 05), as well as plasma concentration of IL - 6 and expression its downstream molecule Tyk2 increased remarkably in patients with acute KD(all P < 0. 05). All the indexes mentioned before restored significantly after IVIG treatment (all P <0. 05). Simultaneously,the 7 former indexes in KD - CAL + group were found to be lower than those in KD - CAL -group,while 2 indexes of the latter in KD - CAL + group were higher than those in KD - CAL - group,and the differ-ences were all statistically significant(all P < 0. 05). Conclusion Histone hypoacetylation of Foxp3 promoter might be one of the important factors contributing to insufficiency and dysfunction of regulatory T cells during acute Kawasaki dis-ease.

OBJECTIVE To study the cardiotoxicity of ophiopogonin D′(OPD′) for rat H9c2 cardio? myocytes. METHODS H9c2 cells were exposed to OPD′ 0.1, 1, 5, 10, 20, 25 and 50 μmol·L-1 for 24 h. Cell viability was examined by MTS assay, and the morphological changes in H9c2 cells were quanti? fied. The cell nucleus injury was examined by high content immune fluorescence screening and the morphological changes were observed under a fluorescence microscope. After treatment with OPD′ 0.1, 1, 5 and 10 μmol·L- 1 for 24 h, the effect on reactive oxygen species (ROS), mitochondrial mem? brane potential(MMP) and apoptosis was detected by flow cytometry. RESULTS The viability was sig? nificantly reduced following exposure to OPD′ 0.1, 1, 5, 10, 20, 25 and 50 μmol·L- 1 (P<0.05,P<0.01). The IC50 value was 9.9 μmol ·L- 1 and cell shrinkage and apoptosis occurred. The levels of ROS and apoptosis rate of H9c2 cells were significantly increased after exposure to OPD′ 0.1, 1, 5 and 10 μmol·L-1 for 24 h (P<0.05,P<0.01) and MMP markedly declined (P<0.05,P<0.01). CONCLUSION OPD′ has significent cytotoxicity on H9c2 cells. It may be related to inducing apopotsis pathways.