In the central nervous system, hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are essential to maintain normal neuronal function. Recent studies have shown that HCN channels may be involved in the pathological process of ischemic brain injury, but the mechanisms remain unclear. Autophagy is activated in cerebral ischemia, but its role in cell death/survival remains controversial. In this study, our results showed that the HCN channel blocker ZD7288 remarkably decreased the percentage of apoptotic neurons and corrected the excessive autophagy induced by oxygen-glucose deprivation followed by reperfusion (OGD/R) in hippocampal HT22 neurons. Furthermore, in the OGD/R group, p-mTOR, p-ULK1 (Ser), and p62 were significantly decreased, while p-ULK1 (Ser), atg5, and beclin1 were remarkably increased. ZD7288 did not change the expression of p-ULK1 (Ser), ULK1 (Ser), p62, Beclin1, and atg5, which are involved in regulating autophagosome formation. Besides, we found that OGD/R induced a significant increase in Cathepsin D expression, but not LAMP-1. Treatment with ZD7288 at 10 μmol/L in the OGD/R group did not change the expression of cathepsin D and LAMP-1. However, chloroquine (CQ), which decreases autophagosome-lysosome fusion, eliminated the correction of excessive autophagy and neuroprotection by ZD7288. Besides, shRNA knockdown of HCN2 channels significantly reduced the accumulation of LC3-II and increased neuron survival in the OGD/R and transient global cerebral ischemia (TGCI) models, and CQ also eliminated the effects of HCN2-shRNA. Furthermore, we found that the percentage of LC3-positive puncta that co-localized with LAMP-1-positive lysosomes decreased in Con-shRNA-transfected HT22 neurons exposed to OGD/R or CQ. In HCN2-shRNA-transfected HT22 neurons, the percentage of LC3-positive puncta that co-localized with LAMP-1-positive lysosomes increased under OGD/R; however, the percentage was significantly decreased by the addition of CQ to HCN2-shRNA-transfected HT22 neurons. The present results demonstrated that blockade of HCN2 channels provides neuroprotection against OGD/R and TGCI by accelerating autophagic degradation attributable to the promotion of autophagosome and lysosome fusion.

BACKGROUND: Pulmonary vein (PV) occlusion generally depends on repetitive contrast agent injection when cryoballoon ablation for atrial fibrillation (AF). The present study was to compare the effect of cryoballoon ablation for AF guided by transesophageal echocardiography (TEE) vs. contrast agent injection. METHODS: Eighty patients with paroxysmal AF (PAF) were enrolled in the study. About 40 patients underwent cryoballoon ablation without TEE (non-TEE group) and the other 40 underwent cryoballoon ablation with TEE for PV occlusion (TEE group). In the TEE group during the procedure, PVs were displayed in 3-dimensional images to guide the balloon to achieve PV occlusion. The patients were followed up at regularly scheduled visits every 2 months. RESULTS: No differences were identified between the groups in regard to the procedure time and cryoablation time for each PV. The fluoroscopy time (6.7 ± 4.2 min vs. 17.9 ± 5.9 min, P < 0.05) and the amount of contrast agent (3.0 ± 5.1 mL vs.18.1 ± 3.4 mL, P < 0.05) in the TEE group were both less than the non-TEE group. At a mean of 13.0 ± 3.3 mon follow-up, success rates were similar between the TEE group and non-TEE group (77.5% vs. 80.0%, P = 0.88). CONCLUSIONS Cryoballoon ablation with TEE for occlusion of the PV is both safe and effective. Less fluoroscopy time and a lower contrast agent load can be achieved with the help of TEE for PV occlusion during procedure.
Aged ; Atrial Fibrillation ; diagnostic imaging ; surgery ; Contrast Media ; Cryosurgery ; methods ; Echocardiography, Three-Dimensional ; methods ; Echocardiography, Transesophageal ; methods ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Veins ; diagnostic imaging ; surgery ; Treatment Outcome

OBJECTIVE: To study the correlation of IL-37 with T lymphocytes subsets and NK cells in ITP patients, and to explore its possible mechanisms involved in the pathogenesis of ITP. METHODS: Forty-five patients with newly diagnosed ITP(newly diagnosed group), 32 patients of complete remission (remission group) and 22 healthy persons(control group) were selected. The serum level of IL-37 in 3 groups was determined by enzyme linked immunosorbent assay (ELISA). The mRNA expression of IL-37, IL-17 and IL-18 in peripheral blood mononuclear cells（PBMNC） in 3 groups was measured by real-time fluorescence quantitative polymerase chain reaction (PCR). The number of IL-18RαCD4 T cells and Tim-3NK cells in the peripheral blood in 3 groups was detected by flow cytometry (FCM). RESULTS: The serum level of IL-37 in the peripheral blood of ITP patients in the newly diagnosed group was significantly higher than that in the control group and the remission group(P＜0.01) . The expression level of IL-37 in PBMNC of the ITP patients in newly diagnosed group was higher than that in the control group and the remission group(P＜0. 05). The expression level of IL-17 and IL-18 in PBMNC of the ITP patients in newly diagnosed group was higher than that in the control group and the remission group(P＜0. 01); the expression of IL-18Rα in CD4 T cells in newly diagnosed group was significantly higher than that in both the control and the remission group(P＜0.01).The expression of Tim-3 in NK cells in ITP patients was significantly lower than that in the control group (P＜0. 01). In ITP patients, the serum IL-37 level and IL-18RαCD4T cells ratio both negatively correlated with Plt count (r=-0.58, r=-0.48) moreo-ver the serum IL-37 level also negatively correlated with amount of CD4 T cells and NK cells (r=-0.29, r=-0.28), but positively correlated with amount of CD8 T cells (r=0.329). CONCLUSION The IL-37 and its receptors may play an immunoregulatory role in CD4 T cells and NK cells, the IL-37 may be a therapeutic target for ITP patients.
CD8-Positive T-Lymphocytes ; Flow Cytometry ; Humans ; Interleukin-1 ; immunology ; Killer Cells, Natural ; Leukocytes, Mononuclear ; Purpura, Thrombocytopenic, Idiopathic ; T-Lymphocyte Subsets

OBJECTIVETo analyze the number of myeloid-derived suppressor cells(MDSC) and the level of prostaglandin E2(PGE2) in the bone marrow of adult ITP patients, and to explore their possible mechanisms involved in the pathogenesis of this disease.

METHODSTwenty-five patients of newly diagnosed ITP, 25 patients of complete remission group and 15 patients of control group were selected. The number of MDSC in the bone marrow between 3 groups was detect by flow cytometry (FCM). The serum level of prostaglandin E2 (PGE2) in 3 groups was determined by enzyme linked immunosorbent assay (ELISA). The relative expression of IFN-γ mRNA in bone marrow mononuclear cells was measured by real time fluorescence quantitative polymerase chain reaction (RT-qPCR) in each groups.

RESULTSThe number of MDSC in the complete remission group was significantly higher than that in the control group (P<0.05); the number of MDSC in the newly diagnosed group was higher than that in the control group; the number of MDSC in the complete remission group was higher than that in the newly diagnosed group. The serum level of PGE2 in bone marrow of ITP patients in the newly diagnosed group was higher than that of the control group（P<0.05）. The serum level of PGE2 in the bone marrow of ITP patients of the complete remission group was higher than that of the control group (P<0.05）. The level of PGE2 in bone marrow serum of ITP patients of the newly diagnosed group was lower than that in the complete remission group(P<0.05）. The relative expression level of IFN-gamma in bone marrow mononuclear cells of the ITP patients in newly diagnosed group was higher than that in the control group and the complete remission group（P<0.001）. The relative quantification (RQ) of IFN-γ in bone marrow mononuclear cells was 2.60 between the newly diagnosed group and the complete remission group.

CONCLUSIONWhen adult ITP disease is remitted, the number of MDSC rises and correlates with the therapeutic response and PGE2 level in the bone marrow.

Adult ; Bone Marrow ; Flow Cytometry ; Humans ; Myeloid-Derived Suppressor Cells ; Purpura, Thrombocytopenic, Idiopathic ; RNA, Messenger

Objective To compare the clinical efficacy and safety of insulin glulisine (GLU) and insulin aspam(ASP) combined with insulin glargine (GLA) in type 2 diabetes patients (T2DM) with secondary failure to oral hypoglycemic agents.Methods One hundred and twenty cases of T2DM with secondary failure to oral hypoglycemia agents were randomly divided into control group (n =60 cases) and treatment group (n =60 cases).The control group received subcutaneous injection of ASP before 0-10 min three meals daily,and the treatment group received subcutaneous injection of GLU before 0-10min three meals daily.The two groups were treated with subcutaneous injection of GLA,once daily.The initial dose of ASP and GLU were 0.8 U · kg-1 · d-1,and the initial dose of GLA was 0.2 U · kg-1 · d-1.Two groups of patients were hospitalized for 2 weeks.The changes of blood glucose,cases and days of blood glucose of achieving target were compared after treatment of hospitalization.The improvement of HbAlc and the frequency of hypoglycemia were observed after treatment of 12 weeks.Results The days of 2-hour postprandial blood glucose(2 h PG) of achieving target in the treatment group and the control group were (10.01 ± 1.99)d and (10.93 ± 1.52)d with 57 cases and 47 cases,and the days of fasting plasma glucose,postprandial blood glucose and 2 h PG of achieving target were (10.31 ± 1.04) d and (11.03 ± 1.38) d with 48 cases and 40 cases(all P ＜ 0.05).After treatment 12 weeks,the HbAlc in treatment and control groups were (6.78 ±0.59)％ and (7.07 ±0.49)％ without significant difference (P ＞ 0.05).During treatment of 12 weeks,the adverse drug reactions in the two groups were mainly hypoglycemic events,the incidence of hypoglycemia in the treatment group was 30.00％,and 25.00％ in the control group,the difference was not statistically significant (P ＞ 0.05).Conclusion Both GLU combined with GLA or ASP combined with GLA have similar efficacy and safety for T2DM with secondary failure to oral hypoglycemic agents.GLU combined with GLA is more rapid in controlling 2 h PG of these patients.

OBJECTIVETo determine the effectiveness and safety of Xinfeng Capsules (XFC) for the treatment of rheumatoid arthritis (RA) patients with decreased pulmonary function.

METHODSThis was a randomized controlled clinical trial of 80 RA patients. Participants were assigned to the trial group (40 cases) and the control group (40 cases) by block randomization. The trial group was treated with XFC, three pills each time three times daily for 2 months. The control group was treated with tripterygium glycoside (TPT), two pills each time three times daily for 2 months. Both groups were followed up after 2 months. The clinical effects, changes in joint and pulmonary function, and quality of life before and after treatment were observed; safety indices were also evaluated.

RESULTSPain, swelling, tenderness, and duration of morning stiffness of joints were obviously decreased after treatment in both the trial and the control groups compared with baseline (P<0.01). Compared with before treatment, hand grip strength increased significantly after treatment in the trial group (P=0.0000); pulmonary function parameters such as forced expiratory volume in the first second of expiration/forced vital capacity (FEV1/FVC), 50% of the expiratory flow of forced vital capacity (FEF50), carbon monoxide diffusing capacity (DLco) were increased (P<0.01 or P<0.05); measures of quality of life such as role-physical, body pain, vitality and mental health were also improved after treatment in the trial group (all P<0.05). Joint swelling in the trial group decreased compared with the control group (P=0.0043), while hand grip strength was increased after treatment (P=0.0000). The increase in FEF50, DLco, and the dimensions of quality of life such as vitality and mental health were all significantly greater in the trial group than the control group (P<0.05 or P<0.01).

CONCLUSIONSXFC not only relieved joint pain in RA patients, but also significantly improved the ventilation and diffusion function of the lungs. Therefore, XFC could improve the whole body function and enhance the quality of life of RA patients.

Adult ; Aged ; Arthritis, Rheumatoid ; blood ; drug therapy ; pathology ; physiopathology ; Blood Sedimentation ; C-Reactive Protein ; Capsules ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Joints ; pathology ; Male ; Middle Aged ; Quality of Life ; Respiratory Function Tests ; Surveys and Questionnaires ; Treatment Outcome

BACKGROUNDAblation of complex fractionated atrial electrograms (CFAE) is an important adjunctive therapy in atrial fibrillation (AF). The present study was to elucidate the substrate underlying CFAE.

METHODSNine adult mongrel dogs were involved in the present study. AF was induced through rapid atrial pacing with vagosympathetic nerve stimulation. CFAE was recorded during AF. Ablation was performed at CFAE sites. Based on the location of the ablation scar, the atrial specimens were divided into CFAE and non-CFAE sites. Serial sections of the atrium were stained respectively with hematoxylin-eosin (HE) and the general neural marker protein gene product 9.5 (PGP9.5). We compared the characteristics of the myocardium and the ganglionated plexus (GPs) distribution between the CFAE and non-CFAE sites.

RESULTSThe myocardium of non-CFAE sites was well-organized with little intercellular substance. However, the myocardium in the CFAE site was disorganized with more interstitial tissue ((61.7 ± 24.3)% vs. (34.1 ± 9.2)%, P < 0.01). GPs in the CFAE site were more abundant than in non-CFAE sites ((34.45 ± 37.46) bundles/cm(2) vs. (6.73 ± 8.22) bundles/cm(2), P < 0.01).

CONCLUSIONThe heterogeneity of the myocardium and GPs distribution may account for the substrate of CFAE and serve as a potential target of ablation.

Animals ; Atrial Fibrillation ; pathology ; Dogs ; Electrophysiologic Techniques, Cardiac ; methods ; Myocardium ; pathology

Objective To determine the predictive value of HATCH score on recurrence of atrial fibrillation (AF) after radiofrequency catheter ablation (RFCA).Methods The data of 123 consecutive AF patients (74 paroxysmal and 49 persistent AF) who underwent RFCA between April 2009 and December 2010 in our department were retrospectively analyzed.Of theses patients,65 (52.9％) patients had HATCH score =0,41 (33.3％) patients had HATCH score =1,and 17 (13.8％) patients had HATCH score ≥2 (HATCH =2 in 11 patients,HATCH =3 in 5 patients,HATCH =4 in 1 patient).The recurrence was defined as atrial tachyarrhythmia lasting more than 30 seconds after 3 months post RFCA.The patients were divided into recurrence group and no recurrence group.Relationship between HATCH score and recurrence was observed.Results There were 43 cases in recurrence group and 80 cases in no recurrence group.After 12 months follow-up,HATCH score was significant higher in recurrence group than in nonrecurrence group[(0.91 ±0.94) score vs.(0.53 ± 0.80) score,P ＜ 0.05].The ratio of patients with HATCH≥2 in recurrence group was higher than in non-recurrence group [23.3％ (10/43) vs.8.8％ (7/80),P ＜ 0.01].The sensitivity and specificity of HATCH ≥ 2 to define the risk of recurrence was 25.0％,92.4％ respectively.Cumulative non-recurrence rate of patients with HATCH score≥2 was lower than patients with HATCH score =0 and 1 (P ＜ 0.05).Conclusion Higher HATCH score is associated with increased risk of AF recurrence post RFCA.

Objective The aim of this study was to investigate the efficiency and safety of ibutilide for cardioversion of persistent atrial fibrillation (AF) during radiofrequency ablation.Methods Eighteen patients (16 males) with persistent atrial fibrillation were enrolled in this study.All patients underwent circumferential pulmonary vein ablation guided by a Carto three-dimensional mapping systen.In addition,linear ablation at the top of the left atrium and the isthmus of mitral valves and complex fractionated atrial electrogram (CAFE) ablation were performed.All patients were still in either atrial fibrillation or atrial flutter after ablation,the patients were treated with l mg intravenous ibutilide injection within 10 minutes after unsuccessful ablation.Intravenous injection was stopped in case of sinus rhythm (SR) restoration or occurrence of severe adverse reactions such as ventricular tachycardia.Cardioversion rate within 30 min and adverse reactions within 4 h were observed.Patients were divided into either conversion group or nonconversion group according to whether AF was converted to sinus rhythm within 30 minutes after injection.Results Eleven patients (61.11％ ) converted to SR after ibutilide injection.There were no significant differences in gender,age,body mass index,left atrium and left ventricular function between conversion group and non-conversion groups.The average conversion time was ( 13.80 + 7.64 ) min,left atrium scar area ratio was significantly larger in non-conversion group ( 12.40 + 11.03 ) ％ than in conversion group (5.12 ±3.83)％,P ＜0.05.Ibutilide significantly prolonged the average wavelength of the AF wave ( 171.8 + 29.5 ) ms vs.(242.0 ± 40.0) ms at baseline,P ＜ 0.01.The QT interval at 30 min after ibutilide injection (0.39 ± 0.21 ) s was significantly longer than before injection (0.51 ± 0.08 ) s,P ＜ 0.05.There was no serious arrhythmias or other adverse reactions post ibutilide injection.Conclusions Ibutilide is highly effective and safe agent for cardioversion in patients underwent unsuccessful ablation.Left atrium scar area ratio is an important determinant for the conversion rate in this cohort.

BACKGROUNDClinical observations have shown that the complex fractionated atrial electrogram (CFAE) associates with ganglionated plexus activity in the cardiac autonomic nervous system. This study aimed to investigate the impact of CFAE ablation on vagal modulation to atria and vulnerability to develop atrial fibrillation (AF).

METHODSTen adult mongrel dogs were involved. Cervical sympathovagal trunks were decentralized and sympathetic effects were blocked. CFAE was color tagged on the atrial 3-dimensional image and ablated during AF induced by S1S2 programmed stimulation plus sympathovagal trunk stimulation. Atrial effective refractory period (ERP) and vulnerability window (VW) of AF were measured on baseline and at vagal stimulation at 4 atrium sites. Serial tissue sections from ablative and control specimens received hematoxylin and eosin staining for microscopic examination.

RESULTSMost CFAE areas were localized at the right superior pulmonary quadrant, distal coronary sinus (CS(d)) quadrant, and proximal coronary sinus (CS(p)) quadrant (21.74%, separately). Sinus rhythm cycle length (SCL) shortening did not decrease significantly after ablation at the sites, including right atrial appendage, left atrial appendage, CS(d), and CS(p) (P > 0.05). ERP shortening during vagal stimulation significantly decreased after ablation (P < 0.01); the VW to vagal stimulation significantly decreased after ablation (P < 0.05). The architecture of individual ganglia altered after ablation.

CONCLUSIONSCFAE has an autonomic basis in dogs. The decreased SCL and ERP shortening to vagal stimulation after CFAE ablation demonstrate that CFAE ablation attenuates vagal modulation to the atria, thereby suppressing AF mediated by enhanced vagal activity. CFAE ablation could suppress AF mediated by enhanced vagal activity.

Animals ; Atrial Fibrillation ; therapy ; Autonomic Nervous System ; Catheter Ablation ; methods ; Dogs ; Electrophysiologic Techniques, Cardiac ; methods ; Electrophysiology ; Female ; Male