Objective:To evaluate the preventive effect of proximal splenic vein ligation after splenectomy on the splenic vein originated portal vein thrombosis (PVT) in portal hypertension.Methods:The clinical data of 94 patients of portal hypertension who had received splenectomy were retrospectively analysed. The proximal splenic vein was ligated in 36 cases during pericardial devascularization and coronary renal shunt with splenectomy. The other 58 cases who had received pericardial devascularization without proximal splenic vein ligation served as control. All of the patients in both groups were given heparin infusion postoperatively through the catheter which was placed in the right gastroepiploic vein during operation. CT portal veinography was performed at the 7th-14th postoperative day for detection of PVT.Results:None of the PVT occurred in the splenic vein ligation group. In the control group, PVT occurred in 22 cases(38%) and splenic vein thrombosis occurred in all the 58 cases (100%). PVT incidence between the two groups is significantly different (0 vs. 38%, χ 2=17.828, P<0.05). Conclusions:Ligation of the proximal splenic vein during splenectomy can effectively prevent the postoperative splenic vein originated PVT in portal hypertension.

Objective To observe the effects of therapy of supplementing spleen to strengthen lung on patients with adenomatous colonic polyps (ACP) after endoscopic removal . Methods One hundred ACP patients with spleen deficiency and dampness blended with blood stasis were randomly divided into 2 groups after endoscopic removal, 50 cases in the treatment group and 50 cases in the control group. Both groups were treated with diet and behavioral therapy, and the treatment group was simultaneously given oral use of modified Shen Ling Baizhu Powder plus self-moxibustion of bilateral acupoint Zusanli point. After treatment for 6 months and 1.5 years, all of the patients were asked to do the examination of colonoscopy, body mass, body mass index (BMI), and triglyceride (TG). Results (1) After treatment, the mean overall symptom scores of the two groups were improved (P < 0.01 compared with those before treatment) , and the improvement of the treatment group was superior to that of the control group(P < 0.01).(2) After treatment for 6 months and 1.5 years, the recurrence rate of ACP in the treatment group was lower than that in the control group(P < 0.05).(3) After treatment for 6 months and 1.5 years, the body mass, BMI and TG of the treatment group were significantly different from those before treatment(P<0.05), while the control group showed no significant changes(P > 0.05). The effect of the treatment group on improving body mass, BMI and TG was better than that of the control group (P < 0.05). Conclusion The therapy of supplementing spleen to strengthen lung is effective on relieving symptoms of ACP patients, and also has an effect on decreasing BMI and TG as well as the recurrence of ACP after endoscopic removal.

ABSTRACT:Objective To investigate the effects of Xiefei Lishui recipe on left ventricle remodeling in rats with heart failure.Methods Heart failure rat model was induced by intraperitoneal injections of doxorubicin. Rats were randomly divided into sham operation group (sham group),model group,traditional Chinese medicine group (TCM group),captopril group,and digoxin group.Distilled water,TCM [22 g/(kg · d)],captopril [19 mg/(kg·d)],and digoxin [32μg/(kg·d)]were administered by gastrogavage in rats in different groups for 35 days,respectively.Indices of ventricle remodeling and cardiac function,plasma levels of B-type natriuretic peptide (BNP),rennin (REN),angiotensin Ⅱ(AngⅡ)and aldosterone (ALD)were measured.Cardiomyocyte apoptosis index and collagen volume fraction (CVF)were analyzed.We also assayed myocardial mRNA expressions of MMP-2/9 and TIMP-1/2,and their tissue inhibiting factors TIMP-1 and TIMP-2.Results Compared with those in sham group,in model group cardiac function was significantly decreased,which could be significantly increased by TCM or captopril or digoxin,indices of cardiac remodeling were significantly increased,which could be significantly decreased by TCM or captopril (P<0.01 or P<0.05).Plasma levels of BNP,REN,AngⅡ and ALD,cardiomyocyte apoptosis index and CVF in model group were significantly increased,could be significantly decreased by TCM or captopril (P<0.01 or P<0.05).Myocardial mRNA expressions of MMP-2,MMP-9,TIMP-1 and TIMP-2 in model group were significantly upregulated compared with those in sham group, which could be significantly downregulated by TCM (P<0.01 or P<0.05).Conclusion Xiefei Lishui recipe can attenuate left ventricle remodeling and improve cardiac function in rats with heart failure, which may be related to downregulating myocardial mRNA expressions of MMP-2 ,MMP-9 ,TIMP-1 and TIMP-2 in the left ventricle as well as inhibiting cardiomyocyte apoptosis and myocardial fibrosis.

Objective: To observe the effect of bisoprolol on cardiac function in heart failure (HF) rats and to explore the mechanism. Methods: The experimental rats were divided into 6 groups: Control group, with normal healthy rats, Sham group, the rats received intraperitoneal injection of normal saline; chronic heart failure (CHF) model was successfully established in 40 rats and divided into 4 groups: CHF group, CHF+bisoprolol (Bis) group, CHF+captopril (Cap) group and CHF+Bis and Cap group.n=10 in each group. The cardiac function was observed among different groups; plasma BNP level was measured by ELISA, myocardial miR-25-3p expression was examined by RT-PCR, protein expressions of SERCA2a and phospholamban (PLB) were detected by Western blot analysis and SERCA2a activity was determined by inorganic phosphorus method. Results: Compared with Control group, CHF group showed decreased cardiac output (CO), left ventricular fractional shortening (LVFS), left ventricular ejection fraction (LVEF), reduced expression of cardiac SERCA2a, PLB, the ratio of SERCA2a/PLB and SERCA2a activity; while increased plasma BNP and miR-25-3p expression, allP<0.01. Compared with CHF group, CHF+Bis, CHF+Cap and CHF+Bis and Cap groups had increased CO, LVFS, LVEF, elevated expression of cardiac SERCA2a, PLB, the ratio of SERCA2a/PLB and SERCA2a activity; while decreased plasma BNP and miR-25-3p expression, allP<0.05.Conclusion: Bisoprolol could improve cardiac function in HF rats, which might be related to down regulating myocardial miR-25-3p expression, up regulating myocardial protein expressions of SERCA2a, PLB and enhancing SERCA2a activity.

Objective To investigate the effects of bisoprolol on myocardial SERCA2a activity in rats with heart fail-ure.Methods Male SD rats were randomly divided into normal control group (control group), sham operation group ( sham group ) , model group , bisoprolol group ( Bis group ) , captopril group ( Cap group ) and bisoprolol plus captopril group[(Bis+Cap)group], heart failure rat model was induced by intraperitoneal injections of doxorubicin .Distilled water, bisoprolol, captopril or bisoprolol plus captopril were administrated by gastrogavage for 35 days, respectively. Indices of cardiac function and plasma levels of B-type natriuretic peptide ( BNP) were measured , myocardial expres-sion of miR-25-3p was detected by Stem-loop RT-qPCR, myocardial levels of SERCA2a and phospholamban (PLB) were detected by Western blot , myocardial SERCA2a activity was determined by the inorganic phosphorus method . Results Cardiac function in model group decreased significantly while plasma levels of BNP were significantly higher than those of control group ( P<0.01 ) .Myocardial expression of miR-25-3p in model group was significantly higher while myocardial levels of SERCA 2a and PLB,SERCA2a activity were significantly lower than those of con-trol group(P<0.01).Cardiac function in Bis group , Cap group and Bis +Cap group improved significantly while plasma levels of BNP were significantly lower than those of model group ( P<0.01 ) .Myocardial expression of miR-25-3p in Bis group, Cap group and Bis +Cap group were significantly lower while myocardial levels of SERCA2a and PLB were significantly higher than those in model group (P<0.01).The SERCA2a/PLB ratio and SERCA2a activity in Bis group and Bis +Cap group were significantly higher than those of model group ( P<0.05 ) .Conclu-sions Bisoprolol therapy improves cardiac function in rats with heart failure , which may be related to inhibition of myocardial miR-25-3p, increasing myocardial SERCA2a and PLB levels, enhancing SERCA2a activity.

AIM:To investigate the effects of Xinkang recipe on myocardial miR-25-3p expression and sarco-plasmic reticulum calcium ATPase 2a ( SERCA2a) activity in heart failure rats .METHODS:Male SD rats were randomly divided into normal group , sham group , model group , Xinkang recipe group ( Xinkang group ) , and captopril group .The heart failure rat model was induced by intraperitoneal injection of doxorubicin .Distilled water , Xinkang recipe and capto-pril were administrated by gastric gavage for 35 d, respectively .The indexes of cardiac function and plasma level of brain natriuretic peptide (BNP) were measured.The SERCA2a activity was determined by the inorganic phosphorus method . The myocardial protein expression of SERCA 2a and phospholamban ( PLB) was detected by Western blot .The myocardial expression of miR-25-3p was detected by stem-loop RT-qPCR.RESULTS:Cardiac output (CO), left ventricular fraction-al shortening ( LVFS) and left ventricular ejection fraction ( LVEF) in Xinkang group and captopril group were significantly higher while the plasma levels of BNP were significantly lower than those in model group (P<0.01).The myocardial ex-pression levels of miR-25-3p in Xinkang group and captopril group were significantly lower while the myocardial protein le -vels of SERCA2a and PLB were significantly higher than those in model group (P<0.01).The SERCA2a/PLB ratio and SERCA2a activity in Xinkang group were significantly higher than those in model group (P<0.05), and no significant change was observed between captopril group and model group .CONCLUSION:Xinkang recipe therapy may improve car-diac function in heart failure rats , which may be related to inhibiting the expression of miR-25-3p, increasing the SER-CA2a/PLB ratio and enhancing SERCA 2a activity in the myocardium .

Objective:To establish an HPLC method to determine the content of schaftoside in Jinshaniu Huashi tablets .Meth-ods:The separation was performed on an Agilent ZORBAX SB-C18 column(150 mm ×4.6 mm, 5 μm)with 0.2% phosphoric acid-acetonitrile as the mobile phase with gradient elution at a flow rate of 1.0 ml· min-1 .The detection wavelength was set at 272 nm, and the column temperature was set at 25 ℃.Results: The calibration curve of schaftoside had a good linearity within the range of 4.58-183.15 μg· ml-1(r=1.000 0).The average recovery was 99.65%with RSD of 1.09%(n=6).Conclusion:The established method is simple and accurate with high repeatability , which can be used for the content determination of schaftoside in Jinshaniu Hua-shi tablets .

Objective To investigate the effects of p300/CBP on histone acetylation of Foxp3 gene and its roles in the immunological pathogenesis of Kawasaki disease (KD).Methods Forty-six children with KD and twenty-eight age-matched health children were consented to participate in this study.Co-immunoprecipitation and real-time PCR were performed to detect Foxp3-associated acetylation levels of histone H4 and binding abilities of p300, CBP, pSmad3 (phosphorylated mothers against decapentaplegic homolog 3) and NF-AT (nuclear factor of activated T cells) with Foxp3 gene in CD4+ T cells.The percentages of CD4+CD25high Foxp3+ cells (Treg) and the expression of Foxp3, CTLA4 (cytotoxic T-lymphocyte-associated protein 4), p300, CBP, TGF-βRⅡ (transforming growth factor β receptor Ⅱ) and pLAT1 at protein level were analyzed by flow cytometry.Quantitative real-time PCR was used to measure the expression of Foxp3, IL-10, TGF-β, TGF-βRⅠ, Egr-1 (early growth response protein 1), RARα (retinoic acid receptor α) and PLCγ1 (phospholipase C-γ1) in Treg cells at mRNA level.Plasma concentrations of TGF-β and retinol acid (RA) were measured by enzyme-linked immunosorbent assay.Results (1) The percentages of Treg cells, levels of Foxp3 and molecules associated with suppressive function of Treg cells (TGF-β, IL-10 and CTLA4), acetylation levels of histone H4 associated with promoter, conserved non-coding DNA sequence 1 (CNS1) and CNS2 of Foxp3 gene decreased remarkably during acute KD (P<0.05), but were restored after IVIG therapy (P<0.05).Meanwhile, all of the aforementioned items in KD patients with coronary artery lesions (KD-CAL+) were lower than those without coronary artery lesions (KD-CAL-) (P<0.05).No significant differences in histone H4 acetylation associated with CNS3 were found among different groups (P>0.05).(2) The levels of p300 and CBP in Treg cells and their binding abilities with Foxp3 gene were down-regulated significantly during acute KD (P<0.05), but were restored to some extent after IVIG treatment (P<0.05).The Foxp3-associated histone acetylation was positively correlated with the expression of p300 and CBP at mRNA level during acute KD (r=0.65, 0.42, P<0.05).Furthermore, the expression of p300 and CBP and their binding abilities with Foxp3 gene in KD-CAL+ group were lower than those in KD-CAL-group (P<0.05).(3) Compared with healthy subjects, plasma concentrations of TGF-β and RA and the expression of TGF-βRⅠ/Ⅱ/Egr-1, RARα and pLAT1/PLCγ1 were down-regulated during acute KD (P<0.05);the binding abilities of pSmad3 and NFAT with Foxp3 gene were reduced remarkably in patients with acute KD (P<0.05).All the items mentioned above were restored after IVIG treatment (P<0.05).Moreover, the ten items aforementioned in KD-CAL+ group were lower than those in KD-CAL-group (P<0.05).(4) Higher acetylation levels of histone H4 associated with promoter, CNS1 and CNS2, and enhanced binding abilities of p300 and CBP with Foxp3 gene were found in CD4+ T cells isolated from patients with acute KD after co-stimulation with TGF-β, RA and anti-CD3/CD28 antibodies as compared with those in CD4+ T cells without stimulation (P<0.05).However, no statistical difference in the acetylation level of histone H4 associated with CNS3 was found between the two groups (P>0.05).Conclusion Hypoacetylation of histone H4 associated with Foxp3 gene caused by insufficient expression of p300/CBP and their impaired binding abilities might be involved with immune dysfunction in KD.IVIG therapy regulates the expression of p300/CBP and their binding abilities with Foxp3 gene through up-regulating TGF-β signal.

Objective To investigate the effects of SMYD3 and MLL5 on histone methylation of Transcription factor forkhead box protein 3 (Foxp3) gene and its roles in the immunological pathogenesis of Kawasaki disease (KD). Methods Forty-two children with KD and 26 age-matched healthy children were consented to participate in this study. Co-Immunoprec-ipitation and real-time polymerase chain reaction (PCR) was performed to determine Foxp3-associated histone methylation levels of H3K4me3 and H3K27me3, and binding levels of SMYD3 and MLL5 with Foxp3 gene in CD4+T cells. The proportion of CD4+CD25high Foxp3+cells (Treg) and protein levels of Foxp3, cytotoxic T lymphocyte associated antigen-4 (CTLA4), TGF-βRⅡand pSmad3 were analyzed by flow cytometry. Quantitative real-time PCR was used to evaluate levels of Foxp3, interleukin (IL)-10, GITR, TGF-βRⅠand RARαmRNA in CD4+T cells. Plasma concentrations of TGF-βand retinol acid (RA) were measured by enzyme-linked Immunosorbent assay. Independent-samples t-test was used as the statistical method in this study. Results ① The proportion of Treg, expression levels of Foxp3 and molecules associated with suppressive function of Treg cells(IL-10, GITR and CTLA4), and histone methylation levels of H3K4me3 associating with promoter, conserved non-coding DNA sequence (CNS) 1 and CNS2 of Foxp3 gene decreased remarkably during acute KD [Promoter:(5.4±1.8)%vs (9.1±2.2)%;CNS1:(2.6±0.9)% vs (3.8±1.1)%; CNS2: (2.4±0.8)% vs (4.2±1.0)%; t=5.50, 6.02, 9.56, 7.92, 7.97, 4.76, 7.73, 5.01, 8.66; P<0.05], and restored after intravenous immunoglobulins (IVIG) therapy [Promoter: (7.2 ±2.1)% vs (5.4 ±1.8)%; CNS1:(3.6±1.4)% vs (2.6±0.9)%; CNS2: (3.6±1.4)% vs (2.4±0.8)%; t=5.56, 4.59, 7.01, 6.04, 5.89, 4.83, 4.45, 4.00, 5.12; P<0.05]. Meanwhile, the nine former items in KD patients with coronary artery lesions (KD-CAL+) were lower than those without coronary artery lesions (KD-CAL-) [Promoter: (4.11±1.45)% vs (6.16±1.93)%; CNS1:(1.99±0.87)%vs (2.96±1.10)%;CNS2: (1.75±0.63)%vs (2.72±1.16)%;t=6.28, 3.24, 4.56, 3.69, 3.38, 4.40, 3.65, 3.00, 3.51; P<0.05]. No significant difference of H3K4me3 associated with CNS3 and H3K27me3 were found among the groups (t=1.03, 0.91, 1.48 and 0.79, 0.82, 1.53; P>0.05). ② Binding levels of SMYD3 and MLL5 with Foxp3 gene in CD4+T cells were down-regulated significantly during acute KD (t=6.63, 6.15; P<0.05), and restored to some extent after IVIG treatment (t=5.36, 4.56; P<0.05). Positive correlations between binding levels of SMYD3 and MLL5 and expression level of Foxp3 mRNA were detected in patients with acute KD (r=0.62、0.45, P<0.05). Furthermore, Binding levels of SMYD3 and MLL5 with Foxp3 gene in KD-CAL+group were lower than those in KD-CAL- group (t=4.11, 4.31; P<0.05). ③ Compared with healthy controls, plasma concentration of TGF-β and RA, and expressions of TGF-βRⅡ, TGF-βRⅠ, pSmad3 and RARα were down-regulated during acute KD (t=11.54, 12.81, 7.43, 16.10, 8.25, 12.06; P<0.05), and elevated remarkably after IVIG treatment (t=8.40, 6.24, 5.94, 11.78, 6.27, 8.30; P<0.05). Simultaneously, all the items aforementioned in KD-CAL+ group were found to be lower than those in KD-CAL-group (t=3.58, 3.30, 3.82, 5.27, 4.71, 3.78; P<0.05). Conclusion Hypomethylation of H3K4me3 associated with Foxp3 gene caused by insufficient binding levels of SMYD3/MLL5 may be involved with immune dysfunction in Kawasaki disease.

Objective To investigate the changes and significances of IL-17-associated histone methylation in the acute phase of Kawasaki disease (KD). Methods Forty-two children with KD and 28 age-matched healthy children were recruited in this study. Co-immunoprecipitation and real-time PCR were performed to detect the IL-17-associated histone methylation in CD4+ T cells. The percentages of CD4+IL-17+ T cells (Th17) and the expression of IL-17 and pSTAT3 at protein level were analyzed by flow cytome-try. Quantitative real-time PCR was used to measure the expression of IL-17, IL-6Rα, gp130, IL-23R, IL-23Rβ1, ETV5, SOCS1, SOCS3, TLR4, MyD88/TRIF, TNFR1 and RIP1 at mRNA level and the expres-sion of miR155 in CD4+ T cells. The levels of IL-6, IL-23 and TNF-αin plasma samples were measured by enzyme-linked immunosorbent assay. Results (1) The children with acute KD showed increased percenta-ges of Th17 cells and enhanced expression of IL-17 and H3K4me3, but inhibited expression of H3K27me3 [H3K4me3:(3.79±1. 45)% vs (1. 93±0. 31)%, H3K27me3: (54. 51±13. 60)% vs (73. 96± 22. 32)%;P<0. 05]. Moreover, the three former indexes in KD patients complicated with coronary artery lesions ( KD-CAL+) were higher than those in KD patients without coronary artery lesions ( KD-CAL-) , while the levels of H3K27me3 in KD-CAL+ group were lower than those in KD-CAL- group [ H3K4me3:(5. 11±1. 68)% vs (2. 98±0. 99)%, H3K27me3:(45. 02±14. 83)% vs (60. 35±12. 51)%;P<0. 05]. A positive correlation was observed between the ratio of H3K4me3/H3K27me3 and IL-17 at transcriptional level in patients with acute KD (r=0. 69, P<0. 05). Treatment with intravenous immunoglobulin (IVIG) restored Th17 cells, expression of IL-17 and methylation levels of H3K4me3 and H3K27me3 to normal levels [H3K4me3:(2. 44 ± 0. 77)% vs (3. 79 ± 1. 45)%, H3K27me3: (66. 52 ± 15. 73)% vs (54. 51 ± 13. 60)%;P<0. 05]. (2) The expressions of pSTAT3, ETV5 and miR155 increased significantly in pa-tients with acute KD, while the expressions of negative regulators of pSTAT3 ( SOCS1 and SOCS3 ) were down-regulated. The expressions of pSTAT3, ETV5 and miR155 in KD-CAL+ group were higher than those in KD-CAL- group (P<0. 05), while the levels of SOCS1 and SOCS3 in KD-CAL+ group were lower than those in KD-CAL- group (P<0. 05). IVIG therapy restored the indexes mentioned above to some extent (P<0. 05). (3) Compared with the healthy subjects, the levels of inflammatory cytokines (IL-6, IL-23 and TNF-α) in plasma and the expressions of surface receptors (TLR4, IL-6Rα/gp130, IL-23R/IL-23Rβ1 and TNFR1) and its downstream adaptors (MyD88, TRIF, RIP1) in CD4+T cells were up-regulated in patients with acute KD (P<0. 05), but were down-regulated significantly after IVIG treatment (P<0. 05). Moreo-ver, all of the indexes mentioned above in KD-CAL+ group were found to be higher than those in KD-CAL-group (P<0. 05). Conclusion Aberrant patterns of IL-17-associated histone methylation might be related to the immune dysfunction in patients with KD.