Objective:To investigate the splanchnic vessel diameter changes following splenectomy in cirrhotic portal hypertension.Methods:The retrospective cohort study was con-ducted. The clinical data of 149 patients with cirrhotic portal hypertension who underwent splenec-tomy in the People′s Hospital of Ningxia Hui Autonomous Region from January 2012 to June 2022 were collected. There were 115 males and 34 females, aged 46(range, 17-68)years. The patients underwent abdominal computed tomography (CT) before surgery and at 1 week, 2 weeks, 1 month, 3 months, 6 months, 1 year, 3 years, 5 years after surgery. The blood vessel diameters in the portal vein system and abdominal visceral artery system were measured after three-dimensional image reconstruction. Measurement data with skewed distribution were represented as M(range). The repeated measured data were analyzed using the generalized linear mixed model, and the marginal mean of the estimated target was expressed as Mean± SE. Simple effects were used to analyze the differences in vascular diameter between pre-surgery and different time points after surgery, and sequential Bonferroni adjusted significance was used for multiple comparisons of marginal means. Results:(1) Changes in diameters of common hepatic artery (CHA), proper hepatic artery (PHA), gastroduodenal artery (GDA) before and after splenectomy. There were significant differences in the diameter change of CHA, PHA and GDA before and after splenectomy ( F=28.66, 29.46, 8.12, P<0.05). The diameter of CHA was thicker at 1 week, 2 weeks, 1 month, 3 months, 6 months, 1 year, 3 years, and 5 years after splenectomy than pre-surgery ( P<0.05). It reached a peak at 1 week after surgery, and then declined slowly and fluctuated. The diameters of PHA and GDA were thicker at 1 week, 2 weeks, 1 month, 3 months, 6 months, 1 year, 3 years, and 5 years after splenectomy ( P<0.05). They reached a peak at 1 month after surgery, and then declined slowly and fluctuated. (2) Changes in diameters of splenic artery (SA), celiac artery (CA), and left gastric artery (LGA) before and after splenectomy. There were significant differences in the diameter change of SA and CA before and after splenectomy ( F=155.33, 66.40, P<0.05). The diameters of SA and CA at 1 week, 2 weeks, 1 month, 3 months, 6 months, 1 year, 3 years, and 5 years after splenectomy were thinner than those before operation ( P<0.05), the changes of which were obvious within 3 months after splenectomy, and then tended to be stable. There was no significant difference in the diameter of the LGA before and after splenectomy ( F=1.07, P>0.05). (3) Changes in diameters of superior mesenteric artery (SMA), right renal artery (RRA), left renal artery (LRA) before and after splenectomy. There were significant differences in the diameter change of SMA, RRA and LRA before and after splenectomy ( F=8.22, 13.21, 10.27, P<0.05). The diameter of SMA at 1 month, 3 months, 6 months, 1 year, and 3 years after splenectomy was thicker than those before operation ( P<0.05). The diameters of both RRA and LRA were thicker at 1 week, 2 weeks, 1 month, 3 months, 6 months, 1 year, 3 years, and 5 years after splenectomy than those before operation ( P<0.05). (4) Changes in diameters of portal vein (PV) and superior mesenteric vein (SMV) before and after splenectomy. There were significant differences in the diameter change of PV and SMV before and after splenectomy ( F=31.74, 2.01, P<0.05). The diameter of PV at 1 week, 2 weeks, 1 month, 3 months, 6 months, 1 year, 3 years, and 5 years after splenectomy was thinner than that before operation ( P<0.05), and there was no significant difference in the diameter of SMV at above time points compared with that before operation ( P>0.05). Conclusions:After splenectomy in patients with cirrhotic portal hypertension, the diameters of PV, SA and CA were reduced, and the diameters of CHA, PHA, GDA and renal artery, superior mesenteric artery were enlarged. These postoperative vessel diameter changes maintain as long as five years.

Objective:To investigate the application value of peri-gastric devasculariza-tion without dissociation of esophagus for portal hypertension.Methods:The retrospective and descriptive study was conducted. The clinical data of 94 patients with portal hypertension who were admitted to three medical centers, including 75 cases in the People′s Hospital of Ningxia Hui Autonomous Region, 12 cases in the People′s Hospital of Wuhai and 7 cases in the People′s Hospital of Wuzhong, from July 2018 to December 2022 were collected. There were 68 males and 26 females, aged 46(range, 21-70)years. All 94 patients underwent peri-gastric devascularization without dissociation of esophagus. Observation indicators: (1) intraoperative condition; (2) postoperative complications; (3) follow-up. Measurement data with normal distribution were represented as Mean± SD, and measure-ment data with skewed distribution were represented as M(range). Count data were described as absolute numbers, and comparison between groups was conducted using the chi-square test or Fisher exact probability. Results:(1) Intraoperative condition. All 94 patients underwent surgery success-fully without operation death, including 82 cases receiving open surgery and 12 cases receiving laparoscopic surgery. The operation time and volume of intraoperative blood loss were (183±85)minutes and 289(range, 158-560)mL, respectively, for the 94 patients. (2) Postoperative complications. Of 94 patients, early portal vein thrombosis occurred in 24 cases, intra-abdominal infection occurred in 2 cases, hepatic encephalopathy occurred in 1 case, pulmonary embolism occurred in 1 case, intra-abdominal hemorrhage requiring operation to stop bleeding occurred in 1 case and pleural effusion requiring drainage occurred in 1 case. All patients with postoperative complications were cured after treatment. None of the 94 patient had postoperative esophageal complications such as odynophagia or dysphagia. (3) Follow-up. All 94 patients were followed up for 38(range, 6-60)months. Of the 45 patients with paraesophageal vein, there were 36 cases of thinner and 9 cases of occlusion of the distal subphrenic paraesophageal vein after surgery, respectively. Cases with esophageal varices disappearance, cases with mild and moderate residual of esophageal varices, cases with severe residual of esophageal varices, cases with recurrence of esophageal varices, cases with esophageal varices bleeding were 7, 70, 9, 4, 4 in the 94 patients after surgery. Cases with esophageal varices disappearance was 7 in the 45 patients with paraesophageal vein, versus 0 in the 49 patients without paraesophageal vein, showing a significant difference between them ( P<0.05). Of 94 patients, 17 cases developed postoperative late portal vein thrombosis and cavernous transformation, 7 cases developed liver cancer, 1 case had hepatic encephalopathy, and 6 cases died. Conclusion:Peri-gastric devascularization without dissociation of esophagus is safe and feasible for the treatment of portal hypertension.

Porcine epidemic diarrhea (PED) is a highly contagious disease that causes high mortality in suckling piglets. Although several licensed inactivated and live attenuated vaccines were widely used, the infection rate remains high due to unsatisfactory protective efficacy. In this study, mRNA vaccine candidates against PED were prepared, and their immunogenicity was evaluated in mice and pregnant sows. The mRNA PED vaccine based on heterodimer of viral receptor binding region (RBD) showed good immunogenicity. It elicited robust humoral and cellular immune responses in mice, and the neutralizing antibody titer reached 1:300 after a single vaccination. Furthermore, it induced neutralizing antibody level similar to that of the inactivated vaccine in pregnant sows. This study developed a new design of PED vaccine based on the mRNA-RBD strategy and demonstrated the potential for clinical application.
Pregnancy ; Swine ; Animals ; Female ; Mice ; Antibodies, Viral ; Swine Diseases/epidemiology* ; Viral Vaccines/genetics* ; Antibodies, Neutralizing ; Vaccines, Attenuated ; Diarrhea/veterinary*

Objective:To study the clinical results of personalized surgical treatment for portal hypertension based on portal venous hemodynamics.Methods:A retrospective study was performed on patients with portal hypertension who underwent surgical treatment from January 2016 to December 2020 at the People’s Hospital of Ningxia Hui Autonomous Region and Wuhai People’s Hospital. Of 229 patients included into this study, there were 156 males and 73 females, with age of (4±11) years old. Portal vein CT and ultrasound doppler examination were performed preoperatively and portal vein manometry and ultrasound doppler examination were performed intraoperatively to evaluate portal venous hemodynamics. Based on the evaluation results, different surgical treatments were adopted. Postoperative complications and results of the operations were recorded. Long-term outcomes were evaluated by the rate of recurrence of gastroesophageal varices which was classified as disappearance, mild, moderate and severe according to endoscopic findings.Results:All the 229 patients completed the operations successfully. All together 13 operative treatments were used: (1) simple splenectomy ( n=11); (2) devascularization ( n=176), including 86 patients with splenectomy combined with extensive devascularization, 44 patients with splenectomy combined with selective devascularization and with preservation of paraesophageal veins, 39 patients with splenectomy combined with selective devascularization and reconstruction of spontaneous portosystemic shunt (34 patients with selective devascularization and reconstruction of spontaneous gastrorenal shunt and 5 patients with selective devascularization and reconstruction of spontaneous splenorenal shunt), 4 patients with secondary devascularization for variceal recurrence and 3 patients with devascularization and preservation of spleen; (3) shunt procedures were performed in 42 patients including 21 patients with splenectomy combined with coronary renal shunt, 11 patients with splenectomy combined with coronary-caval shunt, 6 patients with distal splenorenal shunt, 2 patients with proximal splenorenal shunt combined with devascularization, 1 patient with right gastroepiploic vein-inferior vena cava shunt and 1 patient with trans-inferior mesenteric vein coronary renal shunt. There were no operative deaths. The Clavien-Dindo grade 3 and above postoperative complication rate was 6.6% (15/229). Two hundred and eight patients were followed up for 6-60 months, with a median follow-up of 38 months. Severe recurrent varices were found in 21 patients (10.1%, 21/208), with 5 patients (2.4%, 5/208) presented with variceal bleeding. The rate of severe varices after selective shunting and selective devascularization by reconstructing the spontaneous portosystemic shunt (4.2%, 3/72) was significantly lower than that of the other devascularization procedures (13.7%, 17/124)(χ 2=4.53, P=0.033). Conclusion:Better clinical results were achieved by selecting the appropriate surgical procedures based on portal venous hemodynamic characteristics of patients. Selective shunting and selective devascularization by reconstructing the spontaneous portosystemic shunts significantly reduced the recurrence rate of severe varies.

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:To investigate the computed tomography (CT) examination anato-mical features and clinical significance of paraesophageal vein (PEV) in portal hypertension.Methods:The retrospective and descriptive study was conducted. The clinical data of 173 patients with portal hypertension who were admitted to the People's Hospital of Ningxia Hui Autonomous Region from January 2018 to June 2021 were collected. There were 124 males and 49 females, aged from 22 to 71 years, with a median age of 47 years. Observation indicators: (1) preoperative CT examinations; (2) surgical situations; (3) follow-up. Follow-up was conducted using outpatient examination to detect surgical effects once every 3 months within postoperative 6 months and once every 6 months after postoperative 6 months. The follow-up was up to June 2021. Measurement data with skewed distribution were represented as M(range) and count data were described as absolute numbers. Results:(1) Preoperative CT examinations. The CT detection rate of PEV in the 173 portal hyper-tension patients was 52.60%(91/173). Of 173 patients, 82 cases were negative with PEV and 91 cases were positive with PEV. Of the 91 patients who were positive with PEV, there were 46 cases with paraesophageal varices, 24 cases with thick PEV, 21 cases with thin PEV, 8 cases without esophageal varices and 83 cases accompanied with esophageal varices. Of the 83 patients who were accom-panied with esophageal varices, there were 44 cases with PEV converged alone with azygos vein or semiazygos vein, 39 cases with paraesophageal varices formed above the diaphragm confluent with esophageal varices into azygos vein. (2) Surgical situations. All the 173 patients underwent surgery successfully, including 8 cases undergoing splenectomy, 86 cases undergoing splenectomy combined with modified complete devascularization, 35 cases undergoing splenectomy combined with spontaneous gastrorenal shunt reconstructing devascularization, 41 cases undergoing splenectomy combined with PEV preserving devascularization and 3 cases undergoing splenectomy combined with PEV ring constriction. None of 173 patients had surgical relative death, 67 cases had complica-tions, including 3 cases undergoing splenectomy, 29 cases undergoing splenectomy combined with modified complete devascularization, 11 cases undergoing splenectomy combined with spontaneous gastrorenal shunt reconstructing devascularization, 23 cases undergoing splenectomy combined with PEV preserving devascularization and 1 case undergoing splenectomy combined with PEV ring constriction underwent complications. (3) Follow-up. Of the 173 patients, 159 cases were followed up for 6 to 42 months, with a median follow-up time of 28 months. In the 7 cases undergoing splenectomy who were followed up, there were 6 cases without esophageal varices and 1 case with recurrence of esophageal varices. In the 79 cases undergoing splenectomy combined with modified complete devascularization who were followed up, there were 5 cases without esophageal varices, 67 cases with mild to moderate residual of esophageal varices, 5 cases with severe residual of esophageal varices, 1 case with recurrence of esophageal varices and 1 case with recurrence of esophageal varices hemorrhage. In the 34 cases undergoing splenectomy combined with sponta-neous gastrorenal shunt reconstructing devascularization who were followed up, there were 7 cases without esophageal varices and 27 cases with mild to moderate residual of esophageal varices. In the 36 cases undergoing splenectomy combined with PEV preserving devascularization who were followed up, there were 4 cases without esophageal varices, 21 cases with mild to moderate residual of esophageal varices, 5 cases with severe residual of esophageal varices, 4 cases with recurrence of esophageal varices and 2 cases with recurrence of esophageal varices hemorrhage. In the 3 cases undergoing splenectomy combined with PEV ring constriction who were followed up, there were 2 cases with mild to moderate residual of esophageal varices, 1 case with severe residual of esophageal varices.Conclusions:The CT detection rate of PEV in portal hypertension patients is >50% and the internal diameter and distribution of blood vessels are different in patients. CT examination anatomical features of PEV can be used to guide the formula-tion of surgical methods.

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 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.