Novel predictors of permanent pacemaker implantation following transcatheter aortic valve replacement

BACKGROUND Conduction and rhythm abnormalities requiring permanent pacemakers(PPM)are short-term complications following transcatheter aortic valve replacement(TAVR),and their clinical outcomes remain conflicting.Potential novel predictors of post-TAVR PPM,like QRS duration,QTc prolongation,and supraventricular arrhythmias,have been poorly studied.AIM To evaluate the effects of baseline nonspecific interventricular conduction delay and supraventricular arrhythmia on post-TAVR PPM requirement and determine the impact of PPM implantation on clinical outcomes.METHODS RESULTS Out of the 357 patients that met inclusion criteria,the mean age was 80 years,188(52.7%)were male,and 57(16%)had a PPM implantation.Baseline demographics,valve type,and cardiovascular risk factors were similar except for type II diabetes mellitus(DM),which was more prevalent in the PPM cohort(59.6%vs 40.7%;P=0.009).The PPM cohort had a significantly higher rate of pre-procedure right bundle branch block,prolonged QRS>120 ms,prolonged QTc>470 ms,and supraventricular arrhythmias.There was a consistently significant increase in the odds ratio(OR)of PPM implantation for every 20 ms increase in the QRS duration above 100 ms:QRS 101-120[OR:2.44;confidence intervals(CI):1.14-5.25;P=0.022],QRS 121-140(OR:3.25;CI:1.32-7.98;P=0.010),QRS 141-160(OR:6.98;CI:3.10-15.61;P<0.001).After model adjustment for baseline risk factors,the OR remained significant for type II DM(aOR:2.16;CI:1.18-3.94;P=0.012),QRS>120(aOR:2.18;CI:1.02-4.66;P=0.045)and marginally significant for supraventricular arrhythmias(aOR:1.82;CI:0.97-3.42;P=0.062).The PPM cohort had a higher adjusted OR of heart failure(HF)hospitalization(aOR:2.2;CI:1.1-4.3;P=0.022)and nonfatal myocardial infarction(MI)(aOR:3.9;CI:1.1-14;P=0.031)without any difference in mortality(aOR:1.1;CI:0.5-2.7;P=0.796)at one year.CONCLUSION Pre-TAVR type II DM and QRS duration>120,regardless of the presence of bundle branch blocks,are predictors of post-TAVR PPM.At 1-year post-TAVR,patients with PPM have higher odds of HF hospitalization and MI.

Safety and efficacy of balloon angioplasty compared to stent-basedstrategies with pulmonary vein stenosis:A systematic review and meta-analysis

BACKGROUND Pulmonary vein stenosis(PVS)is an uncommon but known cause of morbidity and mortality in adults and children and can be managed with percutaneous revascularization strategies of pulmonary vein balloon angioplasty(PBA)or pulmonary vein stent implantation(PSI).AIM To study the safety and efficacy outcomes of PBA vs PSI in all patient categories with PVS.METHODS We performed a literature search of all studies comparing outcomes of patients evaluated by PBA vs PSI for PVS.We selected all published studies comparing PBA vs PSI for PVS with reported outcomes of restenosis and procedure-related complications in all patient categories.In adults,PVS following atrial fibrillation ablation and in children PVS related to congenital etiology or post-procedural PVS following total or partial anomalous pulmonary venous return repair were included.The patient-centered outcomes were risk of restenosis requiring re-intervention and procedural-related complications.The metaanalysis was performed by computing odds ratios(ORs)using the random effects model based on underlying statistical heterogeneity.RESULTS Eight observational studies treating 768 severe PVS in 487 patients met our inclusion criteria.The age range of patients was 6 months to 70 years and 67%were males.The primary outcome of the re-stenosis requiring re-intervention occurred in 196 of 325 veins in the PBA group and 111 of 443 veins in the PSI group.Compared to PSI,PBA was associated with a significantly increased risk of re-stenosis(OR 2.91,95%CI:1.15-7.37,P=0.025,I2=79.2%).Secondary outcomes of the procedurerelated complications occurred in 7 of 122 patients in the PBA group and 6 of 69 in the PSI group.There were no statistically significant differences in the safety outcomes between the two groups(OR:0.94,95%CI:0.23-3.76,P=0.929,I^(2)=0.0%).CONCLUSION Across all patient categories with PVS,PSI is associated with reduced risk of re-intervention and is as safe as PBA and should be considered first-line therapy for PVS.

Anatomic Correlates of Mitral Systolic Anterior Motion in Transposition of the Great Arteries Following Atrial Switch Operation

Introduction:We sought to investigate whether the development of sub-pulmonic systolic anterior motion(SAM)may be inherent to the anatomy of the the mitral valve(MV)or affected by external factors,such as a dilated right ventricle or chest abnormalities in d-looped transposition of the great arteries post atrial switch operation(d-TGA/AtS).Methods:Analysis was performed of clinical and cardiac imaging studies acquired on 19 adult patients with d-TGA/AtS(age 42±6 years old,56%male)between 2015–2019.Echocardiography data included mitral apparatus anatomy,and CT/MRI data included biventricular dimensions,function,and Haller index(HI)for pectus deformity.Results:Patients with leaflet SAM(n=6)compared to patients without SAM(n=13)had higher MV protrusion height(2.3±0.5 vs.1.5±0.4 cm,p≤0.01)and longer anterior MV leaflet length(3.1±0.4 cm vs.2.6±0.3 cm p≤0.05),when compared to those without.CT/MRI showed higher sub-pulmonic left ventricular ejection fraction(LVEF)in the SAM group(71%±8%vs.54%±7%,respectively).RV size and function,significant chest deformity(HI>3.5),presence of a ventricular lead pacemaker,and septal thickness did not play a role in development of SAM.Conclusions:An elongated mitral apparatus is associated with the development of SAM,and the development of left ventricular outflow tract obstruction(LVOTO),in d-TGA/AtS.LV hyperkinesia is associated with SAM.Systemic RV dimensions,septal thickness,and degree of chest deformity did not differ significantly between subjects with SAM and those without.

Prediction of permanent pacemaker implantation after transcatheter aortic valve replacement:The role of machine learning

BACKGROUND Atrioventricular block requiring permanent pacemaker(PPM)implantation is an important complication of transcatheter aortic valve replacement(TAVR).Application of machine learning could potentially be used to predict preprocedural risk for PPM.AIM To apply machine learning to be used to predict pre-procedural risk for PPM.METHODS A retrospective study of 1200 patients who underwent TAVR(January 2014-December 2017)was performed.964 patients without prior PPM were included for a 30-d analysis and 657 patients without PPM requirement through 30 d were included for a 1-year analysis.After the exclusion of variables with near-zero variance or≥50%missing data,167 variables were included in the random forest gradient boosting algorithm(GBM)optimized using 5-fold cross-validations repeated 10 times.The receiver operator curve(ROC)for the GBM model and PPM risk score models were calculated to predict the risk of PPM at 30 d and 1 year.RESULTS Of 964 patients included in the 30-d analysis without prior PPM,19.6%required PPM post-TAVR.The mean age of patients was 80.9±8.7 years.42.1%were female.Of 657 patients included in the 1-year analysis,the mean age of the patients was 80.7±8.2.Of those,42.6%of patients were female and 26.7%required PPM at 1-year post-TAVR.The area under ROC to predict 30-d and 1-year risk of PPM for the GBM model(0.66 and 0.72)was superior to that of the PPM risk score(0.55 and 0.54)with a P value<0.001.CONCLUSION The GBM model has good discrimination and calibration in identifying patients at high risk of PPM post-TAVR.

Treatment of coronary in-stent restenosis： a systematic review

冠的 stent 培植显著地改进了经皮的冠的干预并且启用了平凡汽球 angioplasty 的早复杂并发症的管理。然而，新复杂并发症伴随了这些改进：从 neointimal 增生产生的 in-stent 狭窄(ISR ) 。在冠的 angioplasty 以后的 ISR 当前是这个方法的主要限制之一，导致 exertional 心绞痛或急性冠的症候群的复发。在赤裸金属的 stent (BMS ) 以后的 ISR 的临床的发生培植是近似 20%-35% 。drug-eluting stents (DES ) 的使用在 ISR 的出现导致了进一步的减少到 5%-10% 。源于控制临床的研究的证据建议 DES 和 drug-eluting 汽球导管(初上舞台的人) 提供临床的最好， angiographic 导致 ISR 的处理。我们为 BMS-ISR 和 DES-ISR 拿了 pathophysiology，诊断和处理选择的系统的评论。我们讨论最近的 randomised 研究，比较为新 biovascular scafolds 和 scafold 狭窄的话题的 BMS 或 DES-ISR 处理，以及使用使用的不同 DES 或初上舞台的人。

Angiotensin Ⅱ promotes NO production, inhibits apoptosis and enhances adhesion potential of bone marrow-derived endothelial progenitor cells

Endothelial 祖先房间(EPC ) 响应织物局部缺血和 endothelial 损害参予出生后的 neovascularization 和 re-endothelialization 的过程。在场的 EPC 的水平被发现了直接与心血管的疾病的结果被联系，并且能被 stimulatory 或禁止的因素调整。给在血管收缩素 II (AngII ) 和心血管的系统之间的靠近的关系，我们在导出的骨头髓(BM ) 的活动调查了 AngII 的效果 EPC。房间被密度坡度 centrifugation 从老鼠的 BM 孤立。AngII 的管理显著地支持了氮的氧化物(没有) 版本，禁止的 EPC apoptosis 和提高的 EPC 粘附潜力。所有被预告的处理与 valsartan 或 L 名字 EPC 上的这些调停 AngII 的效果稀释。而且， LY294002 和 wortmannin 废除了 AngII 的 anti-apoptotic 效果。西方的污点分析显示了没有 synthase (eNOS ) 蛋白质和 phosphorylated Akt 在 EPC 与 AngII 的处理增加了的那 endothelial。因此， AngII 通过 AngII 类型 1 受体(AT1R ) 改进了 EPC 的几项活动，它可以代表与 angiogenesis 连接 AngII 和 AT1R 的可能的机制。另外，导致 AngII 通过在 EPC 的 eNOS 的合成都不调整 apoptosis 和粘附，并且 PI3-kinase/Akt 小径在导致 AngII 的 antiapoptosis 发信号有一个必要角色。

Management of adults with coarctation of aorta

Coarctation of the aorta(CoA)is a relatively common congenital cardiac defect often causing few symptoms and therefore can be challenging to diagnose.The hallmark finding on physical examination is upper extremity hypertension,and for this reason,CoA should be considered in any young hypertensive patient,justifying measurement of lower extremity blood pressure at least once in these individuals.The presence of a significant pressure gradient between the arms and legs is highly suggestive of the diagnosis.Early diagnosis and treatment are important as long-term data consistently demonstrate that patients with CoA have a reduced life expectancy and increased risk of cardiovascular complications.Surgical repair has traditionally been the mainstay of therapy for correction,although advances in endovascular technology with covered stents or stent grafts permit nonsurgical approaches for the management of older children and adults with native CoA and complications.Persistent hypertension and vascular dysfunction can lead to an increased risk of coronary disease,which,remains the greatest cause of long-term mortality.Thus,blood pressure control and periodic reassessment with transthoracic echocardiography and threedimensional imaging(computed tomography or cardiac magnetic resonance)for should be performed regularly as cardiovascular complications may occur decades after the intervention.

Effects of Kanlijian (坎离煎) on Exercise Tolerance, Quality of Life, and Frequency of Heart Failure Aggravation in Patients with Chronic Heart Failure

Objective： To observe the effects of conventional therapy combined with Kanlijian （ 坎离煎, KLJ） on exercise tolerance, quality of life and frequency of heart failure aggravation in patients with chronic heart failure（CHF）. Methods： Sixty CHF patients differentiated as sufferring from the syndrome of Xin-Shen Yang deficiency were included in the study and randomly assigned at the ratio of 2 ： 1 into the KLJ group （n =39） and the control group（n = 21）. All the patients were treated with conventional therapy of Western medicine, but to those in the KLJ group, KLJ was medicated additionally one dose daily with 24 wks as one therapeutic course. The efficacy on TCM syndrome and changes of scores on TCM syndrome were observed after treatment. The indexes, including 6-minute walking distance （6MWD）, quality of life （QOL, accessed by LHFQ scoring）, NYHA grade, hemodynamic indexes and reducing/withdrawal rate of diuretic and digoxin before and after treatment were recorded and compared. Also the frequency of re-admission due to aggravation of heart failure in one year＇ s time were observed. Results： （ 1 ） The efficacy on TCM syndrome, improvement on scores of TCM syndrome, therapeutic effects on 6MWD, QOL, and NYHA grade in the KLJ group were superior to those in the control group. （2） Hemodynamic indexes after treatment, left ventricular fractional shortening （LVFS） and E peak/A peak （E/A）, between the two groups had no significant difference, while left ventricular ejection fraction （LVEF） was increased significantly in the KLJ group, but with no obvious change in the control group. （3） The reducing/withdrawal rate of diuretic and digoxin in the KLJ group was significantly higher than that in the control group. （4） The 1-year frequency of re-admission significantly decreased in the KLJ group. Conclusion： The adjuvant treatment of KLJ on the basis of Western conventional therapy can significantly improve CHF patients＇ exercise tolerance, quality of life and cardiac function, reduce the dosage of diuretic and digoxin needed, and decrease the re-admission frequency due to aggravation of heart failure.

Expression of NF-κ B and osteopontin of synovial fluid of patients with knee osteoarthritis

Objective:To explore the significance of osteopontin and nuclear factorκB(NF-κB) expression in patients with knee osteoarthritis.Methods:RT-PCR and enzyme-linked immunosorbent assay were used to measure the Osteopontin(OPN) and NF-κB concentration of knee joint synovial fluid of patients with knee osteoarthritis and trauma fractures,and analyze the relationship between the expressiones of them.Results:OPN and NF-κB expression at the mRNA and protein levels of patients with knee osteoarthritis were significantly higher than the control group, the result showed statistical significance(P【0.05).There was a positive correlation between the OPN levels in synovial fluid of patients with knee osteoarthritis and NF-κB expression levels (P【0.05).Conclusions:The high expression of OPN and NF-κB are closely related to occurrence and development of knee osteoarthritis.

Validation of the China-PAR Equations for Cardio-cerebrovascular Risk Prediction in the Inner Mongolian Population

The objective of this study was to evaluate the usefulness of the China-PAR equations in predicting the 10-year risk of cardiovascular disease（CVD） in the Inner Mongolians population. A population-based, prospective cohort of 2,589 Mongolians were followed up from 2003 to 2012.Participants were categorized into 4 subgroups according to their 10-year CVD risks calculated using the China-PAR equations： 〈 5%, 5%-9.9%,10%-19.9%, and ≥ 20%. The China-PAR equations discriminated well with good C statistics（range,0.76-0.86）. The adjusted hazard ratios for CVD showed an increasing trend among the 4 subgroups（P for trend ＆lt; 0.01）. However, the China-PAR equations underestimated the 10-year CVD risk in Mongolians, and the calibration was unsatisfactory（Hosmer-Lemeshow χ^2 = 19.98, P ＆lt; 0.01 for men, χ^2 = 46.58, P 〈 0.001 for women）. The performance of the China-PAR equations warrants further validation in other ethnic groups in China.

Construction of shRNA Targeted to the Rat Angiotensin Ⅱ Type 1 Receptors and Its RNAi in Cytoplasma

The expression vector of shRNA targeted to the rat angiotensin Ⅱ receptor gene was constructed and the efficacy of siRNAs to modulate the expression of target gene in the in vitro cultured mammalian cells was investigated for antihypertensive therapy in spontaneous hypertensive rat （SHR） at post-transcriptional level. The sense and antisense RNA oligonucleotides strands targeting angiotensin Ⅱ receptor mRNA were synthesized individually according to the sequence of the rat angiotensin Ⅱ receptor. For preparation of duplexes, sense- and antisense-stranded oligonucleotides were mixed and annealed, and the annealed duplexes were cloned into the pGenesil-1 vector. The rat glioma cells were transfected with constructed pGenesil-1-shRNA plasmid and scrambled plasmid. The cultured cells were collected at different phases. RT-PCR and Western blot were performed. The AT1 mRNA and protein levels behaved ultimately same. Compared to control after 48 h, AT1 mRNA levels were decreased to 35.5%±3.0 %, and the levels reached their lowest point after 72 h （20.7% ±4 % of control）. At 24 and 48 h, AT1 protein was reduced to 46.9%±4.2% and 36.98%±3.7% respectively compared to control and a maximum reduction was observed after 72 h of incubation （28.1%± 4% compared to controls）. Plasmid-based shRNA expression systems targeted against the rat angiotensin Ⅱ receptor gene were generated successfully. The shRNAs with a 22-nt stem and a short loop were cleaved into small interfering dsRNA （siRNA） by the Dicer. The in vitro transcribed siRNA enables the effective silencing of gene expression to the target mRNA and leads to effective inhibition of translation of proteins and will be lay the foundation of application of gene silencing technology to hypertensive rats.

Proteomic analysis of the serum in patients with idiopathic pulmonary arterial hypertension

Idiopathic pulmonary arterial hypertension(IPAH) is a rare disease of unknown etiology.The exact pathogenesis of pulmonary arterial hypertension is still not well known.In the past decades,many protein molecules have been found to be in-volved in the development of IPAH.With proteomic techniques,profiling of human plasma proteome becomes more feasible in searching for disease-related markers.In present study,we showed the protein expression profiles of the serum of IPAH and healthy controls after depleting a few high-abundant proteins in serum.Thirteen spots had changed significantly in IPAH com-pared with healthy controls and were identified by LC-MS/MS.Alpha-1-antitrypsin and vitronectin were down-regulated in IPAH and may be valuable candidates for further explorations of their roles in the development of IPAH.

Abnormal Calcium “Sparks” in Cardiomyocytes of Post-myocardial Infarction Heart

In ischemic hypertrophic myocardium, contractile dysfunction can be attributed to the decreased calcium induced calcium release （CICR） in cytoplasm. This study aimed to investigate the electrophysiological properties and the expression of L calcium channel subunits in post-MI myocardium. The ischemic heart remodeling model was established in SD rats. The expressions of calcium channel subunits were determined by realtime RT-PCR. Whole cell patch clamp was used to record the electrophysiological properties of L calcium channel. The results showed that the L calcium channel agonist Bayk 8644 induced the significantly decreased CICR in the rat cardiomyocyte 6 weeks after myocardial infarction （MI）. In the post-MI cardiomyocytes, the amplitude of ICaL decreased dramatically and the inactivation curve of the current shifted to more negative potential. At mRNA level, the expression of the calcium channel alpha1c, beta2c subunits decreased dramatically in the ventricle of post-MI rats. The expression of alpha2/delta subunit, however, remained constant. It is concluded that the abnormal expression of the L calcium channel subunits in post-MI cardiomyocytes contributes to the ICaL decrease at early stage of the ischemic remodeling in cardiomyocytes, which leads to the decreased CICR in the cell and contractile dysfunction of myocardium.

Domestic versus imported drug-eluting stents for the treatment of patients with acute coronary syndrome

BACKGROUND:The application of coronary stents,especially drug-eluting stents(DESs),has made percutaneous coronary intervention(PCI) one of important therapeutic methods for CHD. DES has reduced the in-stent restenosis to 5%–9% and signifi cantly improved the long-term prognosis of patients with CHD. The study aimed to investigate the long-term eff icacy and safety of domestic drugeluting stents(DESs) in patients with acute coronary syndrome(ACS).METHODS:All patients with ACS who had undergone successful percutaneous coronary intervention(PCI) in the First Aff iliated Hospital of Zhengzhou University from July 2009 to December 2010 were included in this study. Patients were excluded from the study if they were implanted with bare metal stents or different stents(domestic and imported DESs) simultaneously. The included patients were divided into two groups according to different stents implanted:domestic DESs and imported DESs.RESULTS:In the 1 683 patients of this study,1 558(92.6%) patients were followed up successfully for an average of(29.1±5.9) months. 130(8.3%) patients had major adverse cardiovascular events(MACEs),including cardiac death in 32(2.1%) patients,recurrent myocardial infarction in 16(1%),and revascularization in 94(6%). The rates of cardiac death,recurrent myocardial infarction,revascularization,in-stent restenosis,stent thrombosis and other MACEs were not signif icantly different between the two groups(all P>0.05). Multivarite logistic regression revealed that diabetes mellitus(OR=1.75,95%CI:1.09–2.82,P=0.021),vascular numbers of PCI(OR=2.16,95%CI:1.22–3.83,P=0.09) and PCI with left main lesion(OR=9.47,95%CI:2.96–30.26,P=0.01) were independent prognostic factors of MACEs. The Kaplan-Meier method revealed that there was no significant difference in cumulative survival rates and survival rates free from clinical events between the two groups(all P>0.05).CONCLUSIONS:The incidences of clinical events and cumulative survival rates are not statistically different between domestic DESs and imported DESs. Domestic DES is effective and safe in the treatment of patients with ACS.

Electrocardiographic markers of cardiac resynchronization therapy response:delayed time to intrinsicoid deflection onset in lateral leads

Cardiac resynchronization therapy(CRT)has emerged as an important intervention for patients with heart failure(HF)with reduced ejection fraction and delayed ventricular activation.In these patients,CRT has demonstrated to improve quality of life,promote reverse left ventricular(LV)remodeling,reduce HF hospitalizations,and extend survival.However,despite advancements in our understanding of CRT,a significant number of patients do not respond to this therapy.Several invasive and non-invasive parameters have been assessed to predict response to CRT,but the electrocardiogram(ECG)has remained as the prevailing screening method albeit with limitations.Ideally,an accurate,simple,and reproducible ECG marker or set of markers would dramatically overcome the current limitations.We describe the clinical utility of an old ECG parameter that can estimate ventricular activation delay:the onset to intrinsicoid deflection(ID).Based on the concept of direct measurement of ventricular activation time(intrinsic deflection onset),time to ID onset measures on the surface ECG the time that the electrical activation time takes to reach the area subtended by the corresponding surface ECG lead.Based on this principle,the time to ID on the lateral leads can estimate the delay activation to the lateral LV wall and can be used as a predictor for CRT response,particularly in patients with non-specific intraventricular conduction delay or in patients with left bundle branch block and QRS<150 ms.The aim of this review is to present the current evidence and potential use of this ECG parameter to estimate LV activation and predict CRT response.

Predictors of recurrent angina in patients with no need for secondary revascularization

BACKGROUND: Approximately 20% to 30% of patients with coronary artery disease (CAD)develop recurrent angina pectoris following successful and complete coronary revascularization utilizingpercutaneous coronary intervention (PCI). We aim to investigate predictors of recurrent angina pectorisin patients who have undergone successful coronary revascularization using PCI, but on repeat coronaryangiography have no need for secondary revascularization.METHODS: The study comprised 3,837 patients with CAD, who were enrolled from January2007 to June 2019. They had undergone successful PCI;some of them redeveloped anginapectoris within one year after the procedure, but on repeat coronary angiography had no need forrevascularization. Thrombolysis in myocardial infarction (TIMI) frame count was used to evaluatethe velocity of coronary blood in the follow-up angiogram. Multivariate logistic regression was usedto investigate risk factors for recurrent angina pectoris. Similarly, predictors of recurrent anginaaccording to the TIMI frame count were assessed using multivariate linear regression.RESULTS: In this retrospective study, 53.5% of patients experienced recurrent angina pectoris.By multivariate logistic regression, the following characteristics were statistically identified as riskfactors for recurrent angina pectoris: female sex, older age, current smoking, low-density lipoproteincholesterol (LDL-C) ≥1.8 mmol/L, and an elevated TIMI frame count (P for all <0.05). Similarly, usingmultivariate linear regression, the statistical risk factors for TIMI frame count included: female sex,older age, diabetes, body mass index (BMI), post-procedural treatment without the inclusion of dualantiplatelet therapy.CONCLUSIONS: Patient characteristics of female sex, older age, diabetes, and elevated BMIare associated with an increased TIMI frame count, coronary microcirculation dysfunction, and recurrentangina pectoris after initially successful PCI. In addition, current smoking and LDL-C ≥1.8 mmol/L arerisk factors for recurrent angina pectoris. In contrast, the treatment with dual antiplatelet therapy isnegatively correlated with a higher TIMI frame count and the risk of recurrent angina pectoris.

Functional Expression of Voltage-Gated Sodium Channels Nav1.5 in Human Breast Caner Cell Line MDA-MB-231

Voltage-gated sodium channels （VGSCs） are known to be involved in the initiation and progression of many malignancies, and the different subtypes of VGSCs play important roles in the metastasis cascade of many tumors. This study investigated the functional expression of Nav1.5 and its effect on invasion behavior of human breast cancer cell line MDA-MB-231. The mRNA and protein expression of Nav1.5 was detected by real time PCR, Western Blot and immunofluorescence. The effects of Nav1.5 on cell proliferation, migration and invasion were respectively assessed by MTT and Transwell. The effects of Nav1.5 on the secretion of matrix metalloproteases （MMPs） by MDA-MB-231 were analyzed by RT-PCR. The over-expressed Nav 1.5 was present on the membrane of MDA-MB-231 cells. The invasion ability in vitro and the MMP-9 mRNA expression were respectively decreased to （47.82±0.53）% and （43.97±0.64）% （P〈0.05） respectively in MDA-MB-23 t cells treated with VGSCs specific inhibitor tetrodotoxin （TTX） by blocking Navl.5 activity. It was concluded that Navl.5 functional expression potentiated the invasive behavior of human breast cancer cell line MDA-MB-231 by increasing the secretion of MMP-9.

Construction of recombinant plasmid pEGFP-C2-L539fs/47 and its expression in HEK293 cells

Objective:To reconstruct pEGFP-C2-L539fs/47,a HERG nonsense mutant in eukaryotic expression plasmid,and observe the fusion protein expressed in HEK293 cells(human embryo kidney cells).Methods:After double digestion of pcDNA3-L539fs/47 and pEGFP-C2-HERG with sbf I and Eco91 I,the small product fragment,from pcDNA3-L539fs/47,was subcloned into the big fragment of pEGFP-C2-HERG under T4 ligase.pEGFP-C2-L539fs/47 was identified by agarose gel electrophoresis and sequencing.pcDNA3-L539fs/47 and pEGFP-C2-L539fs/47 were transiently transfected into HEK293 cells by Lipofect,respectively.The expression of fusion protein in HEK293 cells was detected through immunofluorescence,laser confocal imaging scanning in vivo,Western blot and PCR.Results:Mutation region cDNA fragment(about 1 kb) and target vector fragment(about 7.2 kb) were ligated after purification and gel recovery.Agarose gel electrophoresis and sequencing successfully demonstrated eukaryotic expression plasmid pEGFP-C2-L539fs/47,constructed approximately 8.2 kb,sequencing consistent with template gene.The transfection efficiency of recombinant plasmid by fluorescence microscopy was more than60%.Western blot analysis detected pcDNA3-L539fs/47 expression of the protein size 60 KD,the expression of pEGFP-C2 fusion protein size of approximately 90 KD.The L539fs/47 gene expression in HEK293 cells was significant by PCR analysis.Confocal laser imaging showed that pEGFP-C2-L539fs/47 protein was successfully expressed in cytoplasm and cytomembrane of HEK293 cells.Conclusion:pEGFP-C2-L539fs/47 containing the HERG gene mutant was successfully constructed by double digestion method and expressed fusion protein in HEK293 cells,which laid a foundation for the further study on L539fs/47.

One Hundred and Seven Middle-Aged and Senile Cases of Coronary Heart Disease with Ventricular Premature Beat Treated by Qing Xin An Shen Fang

The therapeutic principle of clearing away the heart-heat to induce tranquilization was adopted in 107 middle-aged and senile patients with ventricular premature beat of coronary heart disease on the basis of pathogenic features of phlegm, heat, blood stasis and deficiency in this condition. Qing Xin An Shen Fang (清心安神方a formula for clearing away the heart-heat to induce tranquilization) was used in the treatment group and compared with mexiletine and Fu Fang Dan Shen Tablets (复方丹参片Compound Salvia Tablets) used in the control group. The results turned to be that the markedly effective rate was 85.1% and the total effective rate 96.3% in the treated group, better than that in the control group, with the former having a good antihypertensive and antihyperlipemic effect and an effect of improving microcirculation and clinical symptoms.

Evaluation of Atrial Septal Defect Using Real-time Three-dimensional Echocardiography:Comparison with Surgical Findings

The present study evaluated the application of three dimensional echocardigraphy （3DE） in the diagnosis of atrial septal defect （ASD） and the measurement of its size by 3DE and compared the size with surgical findings. Two-dimensional and real-time three dimensional echocardiography （RT3DE） was performed in 26 patients with atrial septal defect, and the echocardiographic data were compared with the surgical findings. Significant correlation was found between defect diameter by RT3DE and that measured during surgery （r=0.77, P〈0.001）. The defect area changed significantly during cardiac cycle. Percentage change in defect size during cardiac cycle ranged from 6%-70%. Our study showed that the size and morphology of atrial septal defect obtained with RT3DE correlate well with surgical findings. Therefore, RT3DE is a feasible and accurate non-invasive imaging tool for assessment of atrial septal size and dynamic changes.