Objective To investigate the effects of dedicator of cytokinesis 1 (Dock180) knockout on proliferation and apoptosis in rat derived H9C2 cardiomyocytes and their mechanisms.Methods A single guide RNA (sgRNA) targeting rat Dock180 gene was designed and constructed using CRISPR/Cas9 system.A plasmid contained above sgRNA was packaged into lentivirus and selected to knockout Dock180 in the cardiomyocytes.A single clone of cardiomyocyte with Dock180 knockout was established.Cardiomyocytes were divided into negative lentivirus group (Cas9, A group), Dock180 knockout group (B group), Cas9 lentivirus hypoxia group (C group), Dock180 knockout hypoxia group (D group).The expression of Dock180 mRNA was examined by RT-PCR, and relevant proteins were detected by Western blot.The cell proliferation rate of the cardiomyocytes was determined by MTT, and the apoptotic rate was measured by flow cytometry.Results Dock180 mRNA and protein were absent in B andD groups.Compared with A and C groups, p-ERK1/2 and Bcl-2 protein expression and cell proliferation rate were lower in B and D groups respectively (P<0.01), while Bax protein expression and cell apoptosis rate were higher in B and D groups respectively (P<0.05, P<0.01);Compared with A group, Dock180 mRNA and protein, p-ERK1/2 and Bcl-2 proteins and cell proliferation rate were reduced, while Bax protein and cell apoptosis rate were increased in C group(P<0.05,P<0.01).Compared with B group, p-ERK1/2 and Bcl-2 proteins and cell proliferation rate were decreased, while Bax protein and cell apoptosis rate were increased in D group(P<0.05,P<0.01).Conclusions Dock180 knockout with CRISPR/Cas9 can inhibit proliferation and promote apoptosis via p-ERK1/2, Bcl-2 and Bax in H9C2 cardiomyocytes.

Objective: To explore the impact of ticagrelor on myocardial microcirculation in patients of non-ST elevation myocardial infarction (NSTEMI) after percutaneous coronary intervention (PCI). Methods: A total of 80 NSTEMI patients treated in our hospital from 2015-03 to 2015-12 were enrolled. All patients received coronary angiography (CAG) and PCI, they were randomly divided into 2 groups: ticagrelor (TA) group and clopidogrel (CA) group, n=40 in each group. The baseline condition, PCI related parameters, plasma levels of SOD, MDA at pre- and 24h, 72h post-PCI were compared, the incidence of major adverse cardiac events (MACE) within 30 days after PCI was also compared between 2 groups. Results: The baseline condition, proportion of infarction-related vessels, average length and diameter of implanted stents were similar between 2 groups, all P>0.05. Compared with CA group, TA group had the less post-operative CTFC in LAD and RCA, P<0.05; while CTFC in LCX was similar between 2 groups, P>0.05. The ratios of acute pre-operative, post-operative TIMI grade 3 and slow flow incidence were similar between 2 groups, P>0.05. The peak values of CK-MB and cTnI in TA group were lower than CA group, P<0.05. Compared with baseline condition, MDA levels at 24h and 72h post-operation were increased in both groups, all P<0.001; compared with CA group, TA group had the lower MDA level at 24h post-operation, P=0.023, MAD was further reduced at 72h post-operation in TA group, P=0.043. Compared with baseline condition, SOD levels at 24h and 72h post-operation were decreased in both groups, all P<0.001; compared with CA group, TA group had the higher SOD level at 24h post-operation, P=0.013, SOD was elevated at 72h post-operation in both groups, the elevation was more obvious in TA group, P=0.049. The incidence of MACE was similar between 2 groups within 30 days of PCI,. Conclusion: Ticagrelor could improve myocardial microcirculation in NSTEMI patients after PCI; it was safe and without obvious adverse effects.

0bjective To establish a standard curve method with more accuracy employed in fluorescence real-time PCR(RT-PCR)as a alternation of the general method.Methods β-actin and KLK11 plasmid DNA for quantitative standard curve were constructed in our study,and Plasmids of β-actin was employed as a internal control.After serial dilution these plasmid were used as DNA standard to obtained slope.Expression of these two genes in malignant prostate cancer cell line LNCAP were tested by real-time PCR,and we analyzed the RT-PCR results with two different methods and compared their accuracy.Results Thestandards curves made from these linear DNA standards showed good linearity (R2=0.991 and 0.992 for β-actin and KLK11 standards graphs),but also displayed a discrepancy in their PCR efficiency(β-actin 123% and KLK11 99%).There were different results after two different stand curve analytical method:the expression of KLK11 mRNA in LNCAP was downregulated in general standard curve method.In the new analytical method,howerer,KLK11 upregnlated for 4.46-fold.And there was a significant difference between aplification efficiency of targt gene and internal control gene(t=4.829,P<0.05).Conclusions Compared with general standard curve method,the new advanced standard curve method described here avoids an error which considers there is identical amplification efficiency between target gene and internal reference gene.It is considered to be a more correct analytical method in fluorescence real-time PCR.

Objective This study was conducted to investigate the clinical outcomes and safety of percutaneous coronary intervention (PCI) to the single-opened vessel lesion among patients with severe left ventricular systolic dysfunction. Methods Twenty-seven patients with severe left ventricular systolic dysfunction (ejection fraction≤35%) undergoing PCI were included. All the patients received PCI only to the single-opened vessel lesion under the conditions of: (1) There were limitations to open chronic total occlusion (CTO);(2) Single-opened vessel lesion was not calcified and tortuous. Clinical outcomes, including success rate of PCI, changes of symptoms in-hospital, brain natriuretic peptide (BNP) and left ventricular ejection fraction (LVEF) pre-and one week post-PCI, the major adverse cardiac events (MACE, including death, myocardial infarction and target vessel revascularization) at 30-days after discharged were observed. Results The success rate of PCI was obtained in all 27 patients(100%), and all the patients received drug eluting stent implantation. The symptoms improvement occurred in all patients and the NYHA class improved from grade Ⅳto grade Ⅲin 22 patients(81.5%) in-hospital. Significant differences were noted in the mean BNP and LVEF between pre-PCI and one week post-PCI, BNP[(2699.6±1104.7) pg/ml vs. (737.0 ± 261.7) pg/ml, P<0.05],LVEF[(26.9±5.7)%vs. (36.0±3.41)%, P<0.05)]. No MACE happened in-hospital and at 30-days follow up. Conclusions PCI only to the single-opened vessel lesion among patients with severe left ventricular systolic dysfunction under the condition of limitations to open CTO is safe and can significantly improve clinical outcomes in-hospital and at 30-days follow up, but it must be emphasized that single-opened vessel lesion not with obvious calcification and tortuosity.

Objective To summarize the clinical experiences of liver transplantation.Methods Of the nine patients, four operation was standard orthotopic liver transplantation,the latter five were the piggyback liver transplantation.The immunosuppressive protocols included methylprednisolone FK506 and mycophenolatemofeti. Meanwhile intravenous antihepatitis B immunoglobulin and Lamivudine were used to prevent hepatitis B recurrence.Results All patients were cured.Conclusion Liver transplantation can be employed for liver disease both cirrhosis and carcinoma as a conventional surgery.It is an effective way for the treatment of no metastatic liver carcinoma.The immunosuppressive protocols included methylprednisolone FK506 and mycophenolatemofeti,it can prevent immune rejection.

Objective: To explore the relationship between coronary tortuosity and coronary microvascular disease (CMVD). Methods: Patients with typical angina symptoms and without serious coronary artery stenosis by coronary angiography were enrolled from June 2014 to December 2016, and CMVD was diagnosed by single photon emission tomography (SPECT). According to the SPECT results, patients were divided to the CMVD group and non-CMVD group. The baseline clinical characteristics, results of coronary angiography were compared between the two groups. The logistic analysis was used to analyze the relationship between coronary tortuosity and CMVD. Result: A total of 117 cases were enrolled, with 69 cases in the CMVD group and 48 cases in the non-CMVD group. No differences were found in gender distribution, age, hypertension, lipid abnormality, hyperuricemia and uses of statins between the two groups (all P>0.05). Incidence of diabetes (78.26%(54/69) vs. 35.42% (17/48) , P<0.05), hs-CRP ((4.29±2.15)mmol/L vs. (2.63±1.20)mmol/L, P<0.001), LDL-C ((2.98±0.96)mmol/L vs. (2.52±0.83)mmol/L, P=0.008) and homocysteine ((13.7±5.61)mmol/L vs. (11.5±4.38)mmol/L, P=0.025) levels were higher in the CMVD group than in the non-CMVD group. The data derived from echocardiographic examination were similar between the two groups. The Corrected TIMI frame counts were higher in the CMVD group than in non-CMVD group (LAD: 31.56±4.92 vs. 27.31±3.75, LCX: 29.47±4.18 vs. 26.62±3.19, RCA: 29.09±5.05 vs. 26.24±3.28, all P<0.001). The incidences of coronary atherosclerosis (76.81% (53/69) vs. 27.08% (13/48) , P<0.001) and coronary tortuosity ( (60.87% (42/69) vs. 33.33% (16/48) , P=0.035) were also higher in the CMVD group than in non-CMVD group. Logistic analysis found that coronary tortuosity (OR=6.111, 95%CI 2.707-13.794, P<0.001), diabetes (OR=6.565, 95%CI 2.883-14.948, P<0.001) and coronary atherosclerosis (OR=8.918, 95%CI 3.822-20.808, P<0.001) were independent risk factors of CMVD. Conclusion Coronary tortuosity, diabetes and coronary atherosclerosis are related to CMVD in this patient cohort.

Objective To use the ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)to authenticate the chemical composition of Morinda citrifolia,and further experiments verify that Morinda citrifolia regulate miR-223-3p/NLRP3 signaling pathway to inhibit the inflammatory response in rats with polycystic ovary syndrome(PCOS).Methods Use UPLC-MS/MS analyze the chemical composition of Morinda citrifolia.SD female rats were randomly divided into three groups:normal control group(n=5),PCOS model group(n=5)and Morinda citrifolia gavage group(n=5).HE staining observe the morphology of ovarian histology,ELISA method detect the expression levels of testosterone(T),luteinizing hormone(LH),follicle stimulating hormone(FSH),interleukin(IL)-1βand IL-18,RT-qPCR detect the mRNA expression of miR-223-3p and NLRP3,and Western blot detect the protein expression of NLRP3.Results UPLC-MS/MS identify 641 chemical composition of Morinda citrifolia,including luteolin,Apigenin,emodin and other composition.Compared with the normal control group,the number of cell layers of ovarian granule in the PCOS model group is reduced,and follicular cystic dilation,atresia follicles increased,T and LH levels increased(P<0.05),FSH levels decreased(P<0.05),IL-18 and IL-1β levels increased(P<0.05),and ovarian tissue miR-223-3p mRNA expression decreased(P<0.01),NLRP3 mRNA and protein expression increased(P<0.01)in PCOS model group.Compared with the PCOS model group,the proportion of follicles at all levels is normal,the number of granule cell layers increased,T and LH levels decreased(P<0.05),FSH levels increased(P<0.05),IL-18 and IL-1β levels decreased(P<0.05),miR-223-3p mRNA expression in ovarian tissue increased((P<0.05),NLRP3 mRNA and protein expression decreased(P<0.05)in Morinda citrifolia gavage group.Conclusion Morinda citrifolia can improve the ovary pathological state of PCOS rats,change the abnormal sex hormones and reduce the levels of inflammatory factors IL-18 and IL-1β.The mechanism maybe regulate the miR-223-3p/NLRP3 signaling pathway to inhibit the inflammatory response in PCOS rats.

In recent years, transesophageal echocardiography has a trend toward miniaturization, so it has great clinical significance and broad clinical application prospect in the management of Cardiac Surgery ICU patient. This paper presents the characteristics of miniaturized transesophageal echocardiography and its clinical application. And we also focused on the contrast between miniaturized transesophageal echocardiography and standard transesophageal echocardiography and transthoracic echocardiography.

OBJECTIVESTo investigate whether the administration of recombinant human B-type natriuretic peptide (rhBNP) before primary percutaneous coronary intervention (PCI) could further limit the infract size, improve left ventricular function, and alleviate cardiac dilation in patients with acute ST-segment elevation myocardial infarction(STEMI).

METHODSA total of 93 consecutive patients presenting chest pain within 12 hours from the onset, suspicious of first STEMI located at anterior wall undergoing primary PCI, were eligible for enrollment and randomly assigned to either rhBNP group (rhBNP administration starting at 5 min before PCI, 1.5 µg/kg bolus intravenous injection followed by 0.007 5-0.03 µg·kg(-1)·min(-1) for up to 120 hours, n=48) or nitroglycerin (NIT) group (NIT treatment starting at 5 min before PCI, 10-100 µg/min intravenous infusion for 120 hours, n=45). Primary PCI was performed in both groups using post-conditioning (PC) technique. TIMI flow grade, corrected TIMI frame count, and TIMI myocardial perfusion grade were compared between the two groups at the time of infarct related artery (IRA) re-patency. The levels of serum creatine kinase MB isoenzyme (CK-MB) and troponin I (TnI) were measured. Echocardiography was performed at baseline 7 days and 6 months later.

RESULTSBaseline characteristics were similar between the two groups. The percentage of TIMI grade 3 and TIMI myocardial perfusion grade 3 after PCI both tended to be higher in rhBNP group than those in NIT group (95.8%(46/48) vs. 86.7%(39/45), P=0.162) and (72.9%(35/48) vs. 62.2%(28/45), P=0.500). The corrected TIMI frame count was significantly decreased in rhBNP group (21.0±8.7 vs. 28.2±14.8, P=0.005). The myocardial infarct size expressed as the AUC of CK-MB ((3 249±1 101) U/L vs. (4 474±1 661)U/L, P=0.010) or AUC of TnI ((3 670±942) µg/L vs. (4 541±1 098) µg/L, P=0.021) was significantly decreased in rhBNP group compared with those in NIT group. At 7 days after primary PCI, the left ventricular ejection fraction (LVEF) tended to be higher (P>0.05), while the E/e' index and wall motion score index (WMSI) ((11.95±3.31 vs. 14.60±4.09, P=0.030) and (1.74±0.17 vs. 2.40±0.55, P<0.001)) were significantly improved in rhBNP group compared with those in NIT group. BNP level was also significantly lower in rhBNP group compared that in NIT group ((68.3±37.8) ng/L vs. (129.4±64.4) ng/L, P<0.001). During 6-month follow-up, LVEF and WMSI were significantly improved in rhBNP group compared those in NIT group(51.7%±12.7% vs. 46.9%±9.6%, P=0.024 and 1.69±0.35 vs. 1.92±0.47, P=0.020).

CONCLUSIONAdministration of rhBNP before PCI with post-conditioning procedure can further improve myocardial perfusion, limit myocardial infarct size, ameliorate cardiac dysfunction and postpone left ventricular early-stage and long-term remodeling in STEMI patients undergoing primary PCI.

Acute Disease ; Creatine Kinase, MB Form ; Echocardiography ; Humans ; Myocardial Infarction ; Natriuretic Peptide, Brain ; Percutaneous Coronary Intervention ; Troponin I ; Ventricular Function, Left

BACKGROUNDTransradial approach catheterization is now widely used in coronary angiography and angioplasty. The ulnar artery, which is one of the two terminal branches of the brachial artery, may be a potential approach for cardiac catheterization. The aim of this study was to evaluate the safety and feasibility of a transulnar approach for coronary catheterization in non-selective patients.

METHODSA total of 535 consecutive patients were randomly assigned to transulnar approach (TUA) group (n = 271) or transradial approach (TRA) group (n = 264) upon arrival at the catheterization laboratory. Allen's test and inverse Allen's test were not routinely performed. Ultrasound-Doppler assessment of the forearm artery was performed before the procedure, two days after the procedure, and 30 days after the procedure. The primary endpoints of study were the rate of successful artery cannulation and the access-site related complications. The secondary endpoints included the number of needle punctures, total time for the procedure, and major adverse cardiac events (MACE).

RESULTSSuccessful puncture of the objective artery was obtained in 91.5% of the patients in the TUA group, and 95.1% of the patients in the TRA group (P > 0.05). There was no significant difference in hematoma complications between the two groups (7.7% vs. 4.2%, P = 0.100). A motor abnormality of the hand was observed in one patient in the TUA group. There were no arteriovenous fistula or pseudoaneurysm observed in our study. Three (1.1%) patients in the TUA group and 8 (3.0%) patients in the TRA group had occlusion of the access artery (P = 0.137), but none of the patients had symptoms or signs of hand ischemia. There were no significant differences in MACE between the two groups during follow-up.

CONCLUSIONThe transulnar approach is an effective and safe technique for coronary catheterization in non-selective patients.

Aged ; Cardiac Catheterization ; methods ; Coronary Angiography ; methods ; Female ; Humans ; Male ; Middle Aged ; Radial Artery ; surgery ; Ulnar Artery ; surgery