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 investigate the correlation between coronary artery calcification(CAC)scores based on computed tomography(CT)on chest and perioperative complications of emergency percutaneous coronary intervention(PCI)of patients with acute ST-segment elevation myocardial infarction(STEMI).Methods:A total of 263 STEMI patients who admitted to the Chest Pain Center of the Department of Cardiovascular Medicine of Baoding NO.1 Central Hospital from March 2020 to July 2021 were selected.All of them underwent CT examination on chest,and were assessed by CAC scores.They were divided into no calcification and mild group(0～2 scores,129 cases),moderate calcification group(3～5 scores,88 cases)and severe calcification group(6-9 scores,46 cases)according to CAC scores.Perioperative complications and major cardiovascular events(MACE)of patients with different degrees of calcification in the follow-up period were analyzed.The differences among different groups were compared.The correlation between CAC scores and perioperative complications of emergency PCI was further analyzed.Results:Compared with patients of no calcification and mild group,the ages of patients of moderate and severe calcification groups were older(x2=45.815,P＜0.05),and the incidences of hypertension,cerebral infarction,diabetes,multi-vessel disease and MACE of moderate and severe calcification groups were significantly higher(x2=6.762,11.071,6.064,25.036,21.694,P＜0.05).There were significant differences in eGFR and NT-ProBNP levels among the 3 groups(F=8.592,Z=20.890,P＜0.05).Compared with the severe calcification group,the incidence of coronary thrombosis was higher in the no calcification and mild group(x2=7.748,P＜0.05).According to logistic regression analysis,the patients with coronary thrombosis,moderate and severe calcification were more likely to have minor complications(OR=4.847,5.280,11.135,P＜0.001).Patients with older age,hypertension or severe calcification of coronary artery were more likely to occur serious complications,and the MACE incidence was higher within 1 year after surgery(OR=1.151,7.982,10.555,21.729,P＜0.05).Conclusion:Patients with moderate and severe calcification lesions who are assessed by CAC scores based on chest CT have a higher incidence of perioperative complications.CAC scores based on chest CT can be used to assess perioperative complications of emergency PCI and MACE within 1 year after surgery.

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 evaluate the efficacy of endovascular treatment for ruptured descending thoracic aortic aneurysm(rDTAA),and to summarize the treatment experience.Methods The clinical data of 8 patients with rDTAA,who were admitted to the Department of Vascular Interventional Surgery,Zhangjiajie Municipal People's Hospital of China to receive treatment between January 2016 and June 2023,were retrospectively analyzed.The patients included 4 males and 4 females,with a median age of 70.5 years(61-78 years).Preoperative computed tomography angiography(CTA)was performed in all the 8 patients,and the diagnosis of rDTAA complicated by hematoma in the thoracic cavity and/or mediastinum was confirmed.Emergency thoracic endovascular aortic repair(TEVAR)was carried out in all the 8 patients.Results Complete isolation of the thoracic descending aortic aneurysm was achieved in 6 patients,and 2 of them received reconstruction of left subclavian artery by using chimney stent technique.The remaining two patients had a small amount of type Ⅰa endoleak,which disappeared after implantation of a cuff stent at the proximal site.During hospitalization,one patient died of septic shock caused by thoracic aortic-esophageal fistula,and 3 patients developed respiratory failure after operation,which was recovered after thoracic drainage,anti-infection medication and ventilator-assisted breathing therapy.Two patients developed postoperative ischemic stroke,one of them presented as grade Ⅲ motor muscle weakness on the affected side accompanied by speech impairment,and the another patient presented as grade Ⅳ motor muscle weakness on the affected side accompanied by speech impairment.During the follow-up period,one patient died of sudden chest pain 7 months after surgery,and another patient who had not received chimney stent implantation developed type Ⅰa endoleak 18 months after operation,which disappeared after implantation of a cuff stent at the proximal site.The remaining 5 patients showed good clinical condition during the follow-up period.Conclusion For the treatment of rDTAA,TEVAR is clinically safe and effective.

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.