Objective To set up a method to determine cytoplasmic free calcium [Ca~(2+)]_(i) in human platelets with flow cytometry.Methods As a calcium indicator,Fluo 3-AM was used to determine the changes,induced by thrombin,of the level of platelet plasma free calcium [Ca~(2+)]_(i) in presence or absence of outside calcium [Ca~(2+)]_(o),in order to elucidate where the platelet plasma free calcium [Ca~(2+)]_(i) mainly come from and it's role in the activity of platelets.Results The geometric mean of fluorescence intensity of the platelet,labeled with Fluo 3-AM,was increased obviously.The concentration of platelet plasma free calcium [Ca~(2+)]_(i) was increased significantly by thrombin in a dose-and [Ca~(2+)]_(o)-dependent manner.Conclusion The increased level of platelet plasma free calcium [Ca~(2+)]_(o),induced by thrombin,mainly comes from [Ca~(2+)]_(o) and partly from the released [Ca~(2+)]_o of intra-platelet probably.

AIM:To study the reversal effects of multidrug resistance by transfecting tumor necrosis factor ?(TNF-?) cDNA and multidrug resistant 1(MDR1) gene antisense RNA into multidrug resistant breast cancer cell line MCF-7/ADR.METHODS:The recombinant vector of enhanced green fluorescent protein(EGFP) with MDR1 antisense RNA and recombinant vector of red fluorescent protein(DsRed2) with TNF-? cDNA were constructed by RT-PCR and DNA recombinant techniques.The recombinant vectors were transfected into multidrug resistant breast cancer cell line MCF-7/ADR.The cell growth curves,cell apoptosis rates,MDR1 gene expression at mRNA and P-gp levels,and the sensitivity to ADR were determined before and after the transfection.RESULTS:After the transfection,cells showed lower growth rate,higher apoptosis rate,lower MDR1 expression at mRNA and P-gp levels,and the sensitivity to ADR increased significantly.CONCLUSION:Transfection of TNF-? cDNA and MDR1 antisense RNA into multidrug resistant breast cancer cells may have good effects on reversal of multidrug resistance.

AIM: To study the effect of integrin ?2 on adhesion of SK-N-SH neuroblastoma cells to collagen. METHODS: Adhesion of the SK-N-SH cells to immobilized collagen was tested with various concentration of Mg~ 2+ , Ca~ 2+ and with 10 ?g/L anti-?2 monoclonal antibody (mAb) 6F1. A_ 570 was detected as adhesion cell numbers. RESULTS: Mg~ 2+ -dependent adhesion of SK-N-SH cells to type I collagen was increased significantly, with peak adhesion at concentration of 1 mmol/L Mg~ 2+ . A_ 570 with or without Mg~ 2+ was 0.59?0.03 and 0.25?0.01 respectively (P

Background and purpose:Thermal ablation (radiofrequency ablation, RFA/microwave ablation, MWA) is the most commonly used minimally invasive technique for the treatment of liver cancer. Real-time shear wave elastography (SWE) is a new type of ultrasonic imaging technology, which was used in our study to monitor thermal ablation of liver cancer. This study aimed to investigate the stiffness change of liver cancer and that of surrounding liver parenchyma before and after thermal ablation, and to evaluate the application of SWE for monitoring thermal ablation for liver cancer.Methods:From Oct. 2014 to Apr. 2015, a total number of 36 patients, with 39 lesions, were treated with RFA or MWA and got complete response. SWE examination was performed before and after ablation. The SWE-mean, SWE-min, SWE-max, SWE-SD of lesions and the surrounding liver parenchyma were measured. Statistical analysis was made to compare the stiffness changes of liver cancer with those of the surrounding liver parenchyma before and after thermal ablation, and to determine whether there were differences between two different ablation modes.Results:Before and after ablation, the SWE-mean of lesions was (30.09±11.67) kPavs (52.11±17.56) kPa,SWE-min was (10.46±8.22) kPavs (20.57±11.42) kPa, SWE-max was (51.50±20.84) kPavs (88.54±27.75) kPa, SWE-SD was (10.63±4.30) kPavs (16.89±7.72) kPa; There were statistically signiifcant differences (P<0.05). Before and after ablation, the SWE-mean of surrounding liver parenchyma was (8.84±2.82) kPavs (8.91±2.78) kPa, SWE-min was (4.77±1.95) kPavs (4.69±1.90) kPa, SWE-max was (13.82±3.79) kPavs (14.34±3.97) kPa, SWE-SD was (3.24±1.32) kPavs (3.37±1.29) kPa; There were no statistically signiifcant differences (P>0.05). After ablation, the SWE-mean of RFA and MWA was (45.55±10.91) kPavs (60.59±20.99) kPa, SWE-min was (18.95±8.86) kPavs (25.93±10.93) kPa, SWE-max was (76.58±15.51) kPavs (104.01±32.59) kPa, SWE-SD was (13.82±3.52) kPavs (20.85±9.77) kPa; There were statistically signiifcant differences (P<0.05).Conclusion:SWE can quantitively analyze the stiffness of lesions. The ablation zone became stiffer after RFA or MWA, and the ablation zone of MWA was stiffer than that of RFA. Two kinds of ablation methods did not signiifcantly affect the stiffness of liver parenchyma around the lesion. SWE could potentially be used to monitor thermal ablation of liver cancer.

Objective To investigate the predictive value of combined analysis on single nucleotide polymorphisms (SNPs) of X-ray cross-complementing1 ( XRCC1 ) gene 194 and 399 codon,xeroderma pigmentosum group D (XPD) gene 312 codon and glutathione S-transferase P1 (GSTP1) gene 105 codon in platinum based chemotherapy.Methods Direct sequencing was performed to detect XRCC1,XPD and GSTP1 genotypes in peripheral blood from 50 cancer patients receiving platinum-based chemotherapy.Genetic polymorphisms of these genes related to sensitivity of platinum were reviewed.Results Favorable genotypes were Arg/Trp and Trp/Trp in XRCC1 194 codon,Arg/Arg in XRCC1 399 codon,Asn/Asn in XPD 312 codon and Val/Val in GSTP1 105 codon.The response rate to chemotherapy was 57.1％,75.0％,60.9％,85.7％ and 87.5％,respectively.The response rate for patients possessing ≥2 favorable genotypes and those possessing 1 or 0 favorable genotype was 78.9％,36.4％ and 0,respectively.Patients possessing ≥2 favorable genotypes demonstrated higher sensitivity to platinum based chemotherapy,compared with those possessing 1 or 0 favorable genotype ( x2 =25.79,P ＜ 0.01 ).Conclusions Combination analysis of genomic polymorphisms of XRCC1,XPD and GSTP1 may be useful in predicting sensitivity of platinum based chemotherapy.

The paper illustrates the role of humanistic multi-element administrate model in the clinical laboratory operation on various aspects such as the construction of the institution,the education of professional personnel and the enhancing of the personnel's enthusiasm.

Objective To compare the mid-term results of video-assisted pulmonary veins isolation and box-lesion for lone atrial fibrillation.Methods Between June 2010 to November 2012,80 lone atrial fibrillation(LAF) patients underwent minimally invasive surgical ablation in Beijing Anzhen Hospital,and all of them received left atrial appendage excision and Marshall ligament break in the surgery.Among them,37 cases were performed two epicardial ablation lines in order to created box-lesion based on PVI(14 paroxysmal AF and 23 persistent AF).Follow-up was finished after discharge.Results Mean follow-up was(18.8 ± 7.4) months,and 2 patients were died with non-cardiac disease.The success rate is 78.8％ (Paroxysmal AF 87.9％;Persistent AF 72.3％).In the paroxysmal AF patients,the success rate for PVI and Box lesion treatment was 84.6％ (11/13) and 92.9％ (13/14),P =0.45;in the persistent AF patients,the success rate for PVI and Box-lesion was 58.3％ (14/24) and 87.5％ (14/16),P =0.04.Conclusion Addition of epicedial ablation lines might increase the cure rate for lone AF therapy,especially for persistent AF.

Objective To conclude the preliminary results of aortic valve repair with autopericardium.Methods After routine median sternotomy the pericardium was isolated and immersed into the 0.6% glutaraldehyde immersion for 10 minutes, then rinsed off before use.Conventional extracorporeal circulation was established and diseased aortic valves were removed.The curves lengths between three commissures measured with the specific sizers.The corresponding size of the pericardial patchs was used to cut the new aortic leaflets.The continuous Johnson Prolene 4-0 sutures were used to fixed the new aortic valve leaflets to the anulus.Another new 4-0 prolene suture were used to fix the new three commissures outside the aortic wall with mat.The new mat stripes were used to fix the sinus in condition with the aortic sinus dilation or bileaflets malformation.Results Since 2015 October total 27 patients received aortic valve repair with autologous pericardium.The age was in the range of 27 to 60 years old including 18 female and 9 male.There were 7 cases diagnosed as aortic valve stenosis and 20 cases aortic insufficiency.And there were 3 cases bileaflets aortic valve deformity.The mean bypass time is(104±25)mins.Postoperative esophageal ultrasound showed good coaptation of the aortic valve without significant regurgitation.The mean transvalve gradient was(8.5±3.2)mmHg(1 mmHg=0.133 kPa).In the early stage one case diagnosed with aortic valve stenosis and regurgitation received monocusp repaired with autologous pericardial leave.But there was still moderate aortic regurgitation and had to receive the aortic valve replacement.All patients recovered well and were treated with Aspirin for 6 months.The followed up lasted for 3 to 13 months.Mean aortic occlusion time was(104±25) minutes.The patients were followed up for 3-13 months, the cardiac function and aortic valve function were stable, and no gradient increased.Conclusion It is relatively simple to perform the aortic valve repair with individual autologous pericardial sizer technique and the primary clinic results are satisfactory.It is worthy of further study and long-term follow-up.

Objective To investigate the clinical results of extracorporeal membrane oxygenation (ECMO) technique during the peri-operative heart transplantation. Methods The clinical data of heart transplantations supported by the ECMO were retrospectively analyzed, including during the bridge to transplant, working as routine bypass in the operation room, and supporting the heart failure after surgery. Results Three 3 cases were supported with ECMO to extend the waiting time: 2cases died of multi organ failure during the waiting, and the third one received the heart transplant and the ECMO was successfully removed. Totally 12 cases of heart transplantation received the modified open style ECMO techniques replacing the routine bypass. The ECMO was removed after assistance for (38 ± 14) h. All the recipients except one death recovered well without right ventricular failure or obvious tricuspid regurgitation which was common after heart transplantation. Ten cases received ECMO support for the heart failure after the transplant. The initial application time ranged from 2 h to 2 days and lasted 43～176 h. One died of infection and two died of multi organ failure nothing to do with ECMO application and other 7 discharged with NYHA Ⅰ/Ⅱ cardiac function. Conclusion Early use of ECMO could safely bridge the heart failure patients to the transplant. Replacing the routine with ECMO techniques in the heart transplantation could get good myocardial protection results even with long cold ischemic time and improve the cardiac function recovery. For the heart failure patients after the surgery the ECMO could afford good support effect.

Objective To evaluate the therapeutic results of percutaneous transhepatic stent angioplasty in patients with portal vein stenosis following liver transplantation.Methods From 2005 to 2013,38 patients developed portal vein stenosis following liver transplantation.Percutaneous transhepatic angioplasty of the portal vein stenosis was performed on these patients.The results were monitored by clinical follow-up and imaging studies.Results Percutaneous transhepatic angioplasty was successful in these patients.Self-expanding metallic stents (n=7),balloon-expandable coronary stent (n=29),and membranous stent (n=1) were used.The follow-up period ranged from 3 to 90 months.Portal venous patency was maintained in 34 patients (one patient died due to multi organ failure,1 patient accepted a third liver transplantation because of biliary tract complication,and 1 patient received a repeat placement of a membranous tent because the portal vein stent was blocked by a tumor thrombus,and 1 patient developed stent restenosis).There was 1 patient who developed hemorrhage in the early postoperative period (2.63％).A diagnosis of hepatic artery hemorrhage was made by hepatic artery angiography and the patient was treated by interventional embolization.Conclusion Percutaneous transhepatic stent angioplasty is an efficacious and safe method to treat portal vein stenosis following liver transplantation.