Objective To study movement process of circulating tumor cells (CTCs) in the blood and mechanism of CTC capture by CellCollector, and reveal relationship between the detected CTC numbers and the actual CTC concentration in the body. Methods Based on Fluent and EDEM software, the unidirectional fluid-solid interaction method was applied to establish a two-phase flow model, including the hemodynamic model and the CTC transport model, and capture simulations under different CTC concentration conditions were conducted. Results The number of CTCs captured by CellCollector was significantly positively correlated with the CTC concentration in the body. When the CTC concentration was low, CTCs could only be captured in several time intervals, and the capture had a certain contingency; as the concentration increased, the uniformity of CTC capture over time became better, and the total number of captures also increased. Conclusions Through the fitting of simulation results, analytical quantitative relationship between the captured CTC number and the CTC concentration in the body is preliminarily given, which provides theoretical basis and mechanical explanation for the clinical use of CellCollector.

Objective To systematically evaluate the clinical outcomes of patients with multivessel diseases treated by fractional flow reserve (FFR) guided percutaneous coronary intervention (PCI) or other forms of treatment .Methods Some keywords inclu-ding FFR or fractional flow reserve ,percutaneous coronary intervention or PCI ,multivessel coronary artery disease were used to search randomized control trails(RCT) ,which compared outcomes of patients with multivessel disease treated by FFR guided PCI with other forms of treatment ,in Chinese and English database including CNKI ,VIP ,Wanfang ,Pubmed ,EMBASE and the web of science .The quality of included studies was evaluated by Jadad quality score and all the data was analyzed by stata 10 .0 .Results A total of 7 english literatures accumulating 2024 cases in experimental group and 7 284 cases in control group were included in this study .Compared with other forms of treatment ,patients with multivessel diseases treated by FFR-guided PCI had significantly low-er risk of myocardial infarction (RR=0 .72 ,P=0 .008) .Although the risk of death and major adverse cardiovascular events were al-so be reduced ,there were no statistically significant difference .Conclusion FFR-guided PCI is an effective treatment for multivessel disease ,but it is still needs further verification for the application in Chinese population .

OBJECTIVETo establish a reliable approach to primary culture and identification of sinoatrial node (SAN) cells.

METHODSThe SAN cells were cultured from SAN tissue removed from neonatal Wistar rats and purified with differential attachment and 5'-bromodeoxyuridine (BrdU) treatment. The obtained cells were morphologically observed with inverted microscopy and transmission electron microscopy. Its action potential was recorded using electrophysiological methods.

RESULTSThree distinctly different cells were observed in the cultured SAN cells: spindle, triangle and irregular. Of these, the spindle cells comprised the greatest proportion, with their shape, structure and electrophysiological characteristics consistent with those of the pacemaker cells of SAN. The triangle cells were similar in features to the similarly shaped myocytes located in the atrial myocardium.

CONCLUSIONSThe culture method of differential attachment combined with BrdU treatment is a reliable approach to growing SAN cells. Of the cells cultured from SAN, the spindle cells appear to function as pacemaker cells.

Animals ; Animals, Newborn ; Cells, Cultured ; Female ; Male ; Microscopy, Electron ; Rats ; Rats, Wistar ; Sinoatrial Node ; cytology ; physiology ; ultrastructure

Objective To develop new methods to cultivate, retrieve and purify mouse mesenchymal stem cells (mMSCs). Methods Bone marrow was collected from 2-month-old Kunming mice by flushing femurs and tibias with complete medium of DMEM-LG. Cells were plated in a Petri dish. After 24 hours, non-adherent cells were removed by two to three washes with PBS, adherent cells were further cultured in complete medium and retrieved by trypsinisation with 0.25% trypsin for 5 min at 37 ℃. The treated adherent cells were cultivated with 3?dilution for further generations. CD11b-negative cells were retrieved from the collected adherent cells of 3rd generation by using immunomagnetic microbeads, and continued to be cultured in complete medium. After the cultured cells were retrieved, their morphology and their ability of osteoblastic differentiation and adipocytic differentiation were examined. Results Most of mMSCs from 1st generation were of shuttle shape, some of irregular shape. After treatment with magnetic microbeads and several generations, mMSCs were of spindle, star and irregular shape. These cells were of rich cytoplasma, clear nucleolus, and grew in parallel or vortex. The cultured adherent cells from the first and subsequent generations had plenty of CD11b-positive blooding-making cells. After 20-day osteoblastic induction, mMSCs differentiated into bone cells, which showed orange phosphate in extracellular matrix by Alizarin red S staining. mMSCs could differentiate into lipocytes. The size of cells increased along with fat-developing induction period. These cells showed many orange fatty follicles with O Red Oil dyeing. Conclusion Pure mMSCs can not be retrieved by either adhering method or generation cultivation method separately. The combined methods of adhering, immunomagnetic microbeads, and serial subcultivation is effective in vitro in retrieve mMSCs.

6 months (n=12), control group, without AF (n=8). Atrial muscle samples of the left and right atrial appendage were cut off during heart valve replacement. Expressions of Cx40 and Cx43 were detected by immunoblotting and immunohistologic analyses. Results ①No obvious change of Cx43 and Cx40 in the left and right atrium of each patient in each group was measured. ②Compared with that in the control group, the level of Cx40 was decreased signficantly in GroupⅠ (P

Objective To evaluate the therapeutic efficacy and safety of transcatheter closure of atrial septal defect (ASD) and patent ductus arteriosus (PDA) using Amplatzer occluder. Methods Routine cardiac catheterization and angiography were performed in 50 patients (23 male, 27 female, age ranging from 3 to 64 years old), including 19 cases of ASD and 31 cases of PDA under local or general anesthesia. After balloon sizing of the ASD, the optimal Amplazter septal occluder (ASO) was transmitted into the left atrial, and the left and right disks were released in turn. The Amplatzer occluder was completely released after transthoracic echocardiography confirmed that there was no residual shunts or new onset mitral valve regurgitation. The Amplatzer duct occluder (ADO) size was selected according to the narrowest point of PDA measured by angiography, and the occluder was released after the repeated angiography showed no residual shunts. Results ① The mean diameter of the ASD measured by balloon was 13-31 (23?6) mm and the diameter of ASO was (17-40) mm. The immediate closure rate was 100%. ② Angiography confirmed that closure of the ductus using ADO was achieved in 30 patients, and closure of the large size (12 mm) was achieved in 1 case of PDA patient using ASO (17 mm). No complications were encountered. Conclusion Transcatheter closure of ASD and PDA using Amplatzer device, with the advantages of simple operation, confirmative occlusion efficacy, minimal invasiveness, wide indications, and less complications, has a bright future of clinical application.

Objective To investigate the effect of calcium/calmodulin-dependent protein kinaseⅡ (CaMK[KG-*6]Ⅱ) inhibitor,KN93 on calcium overloading of atrial muscle cells in neonatal rats and detect the expression of CaMK[KG-*6]Ⅱ. Methods The atrial muscle cells from neonatal rats were primarily cultured for 96 h and then divided into 6 group,control,calcium overloading group,KN93 group (0.5 ?mol/L),low-,moderate-and high-dose of KN93+ calcium overloading group. A model of calcium overloading for atrial muscle cells was established by using calcium ionophore (ionomycin,1.0 ?mol/L). For the later 3 groups,KN93 at doses of 0.25,0.5 and 1.0 ?mol/L was added into the culture medium for 30 min followed by 1.0 ?mol/L ionomycin treatment for another 30 min. The identification of ?-actin was performed by immunofluorescence staining. In the present of Fluo-3/AM (an indicator of calcium),intracellular calcium and the expression of CaMK[KG-*6]Ⅱ were detected under the intervention of KN93 with laser cofocal microscopy and Western blotting respectively. Results More than 90% of cultured cells were positive to ?-actin antibody. Compared with the control group,the fluorescence intensity of intracellular Ca2+ was increased significantly by ionomycin (660.16?108.47 vs 376.12?57.57,P

AIM: To study influence of ischemia-reperfusion(IR) on apoptosis and expression of apoptosis-related genes Fas-L, Bax and Bcl-2 of sinoatrial node(SAN) cells in rabbits in vivo. METHODS:Ninety healthy adult rabbits were divided randomly into control group, ischemia groups (I 10 min , I 30 min , I 60 min and I 120 min ) and IR groups (I 10 min R 4h , I 30 min R 4h , I 60 min R 4h and I 120 min R 4h ). IR injury model of SAN was established by occluding and loosening the start section of right coronary artery. The apoptosis of SAN cells was detected by TUNEL staining. The expression of Fas-L, Bax and Bcl-2 of SAN cells was detected by immunohistochemistry. RESULTS:①No obvious apoptosis of SAN cells was observed in control group, I 10 min and I 30 min groups. Apoptosis of different degrees in SAN cells were found in 68.3%(41/60) rabbits in I 60 min , I 120 min and 4 subgroups of IR. ②The highest expression of Fas-L and Bax was observed in I 120 min group and that of Bcl-2 was in I 60 min group. ③The highest expression of Fas-L and Bax was observed in I 60 min R 4h group. The peak level of Bcl-2 was observed in I 30 min R 4h group. ④The expression of Fas-L and Bax was significant higher in IR group than that in ischemic group at the same time point. CONCLUSION:Ischemia and IR induced apoptosis of SAN cells in rabbit in vivo . Fas-L、Bax、Bcl-2 may participate in the regulation of apoptosis and the injury during IR aggravates the apoptosis of SAN cells.

AIM: To study the effects of ischemia/reperfusion (I/R) on primary cultured sinoatrial node (SAN) cells and the influence of pinacidil (a K_(ATP) channel activator). METHODS: The SAN cells were isolated from newborn rats and purified. The 48 h cultured cells were cultivated in following mediums: simulated reperfusion solution as normal control, simulated ischemia/reperfusion solution (I/R), Pinacidil+I/R (P+I/R), 5-HD+P+I/R and 5-HD+I/R. Spontaneous action potentials were recorded by ruptured-patch whole-cell technique in current clamp ((I=0)) and the maximum diastolic potential (MDP), upstroke velocity (UV), action potential overshoot (APO), interbeat interval (IBI) and action potential durations at 50% repolarization (APD_(50)) were measured. RESULTS: Compared with control group, simulated ischemia/reperfusion shorten APD_(50), reduced UV, MDP and APO. Exposed to pinacidil, MDP of cells in I/R groups was hyperpolarized; IBI, UV and APO were increased; APD_(50) was shorten. 5-HD couldn't block the effects of pinacdil on APD_(50), IBI and MDP, but reversed its actions on increasing UV and APO. CONCLUSIONS: Pinacidil made changes of AP in I/R group by opening different K_(ATP) channels of SAN cells. The role of this changes on protection in SAN cells during ischemia/reperfusion requires further investigation. [

Objective To study the effects of mimic ischemic preconditioning (IP) on i and I-LCa of sinoatrial node cells and explore the protective effects of IP. Methods Cells were randomized to three groups: control, ischemia/reperfusion (I/R), IP. After labeled with fura4, fluorescence intensity of i was studied with laser confocal microscopy and I-LCa was recorded by whole-cell patch clamp technology. Results IP significantly reduced the fluorescence intensity of i and enhanced the peak of I-LCa as compared with I/R, shifted the I-V curve to more negative value. Conclusion IP can reduce the overload of i caused by I/R and increase I-LCa weakened by I/R.