Objective To establish an acurate and convenient method to distinguish human platelet antigen(HPA) SNPs based on Target Enriched Multiplex-PCR(TEM-PCR),fluorescent probe melting curve analysis and blood direct PCR.Methods Design TEM-PCR primers and probes of HPA1-17,Cab alleles,amplify target sequences of all 18 alleles by blood direct PCR and distinguish different SNPs by melting curve of probes.Results The TEM-PCR could amplify all target sequences of 18 alleles and the melting curve analysis could distinguish those SNPs,the accuracy was equal to PCR-SSP method and the process was more convenient without blood genomic DNA extraction and subsequent gel electrophoresis thus decrease the cross-contamination risk.Conclusion Successfully established a HPA1-17,Cab alleles distinguishing method based on TEM-PCR,blood direct PCR and fluorescent probe melting curve analysis technique.

Objective To evaluate the risk factors of Sun' s procedure for acute Stanford A type aortic dissection involving aortic arch.Methods Between February 2009 to February 2012,data from 233 patients who underwent Sun' s procedure for acute Stanford type A aortic dissection involving aortic arch were collected retrospectively.All the risk factors related to mortality were analyzed by univariate statistical analysis.Significant univariate variables were entered into multiple logistic analysis.Results Total 23 patients died in the hospital and the mortality was 9.87 ％.Univariate analysis showed that age,limb ischemia,and cardiopulmonary bypass longer than 268 minutes were risk factors for mortality.After these factors were entered into multiple logistic regression analysis,multiple logistic regression showed that age(P =0.017,OR =1.062),cardiopulmonary by pass time longer than 268 minutes(P =0.001,OR =6.150) were final independent risk factors for mortality.ConclusionAge and cardiopulmonary bypass time longer than 268 minutes were final independent risk factors for mortality.Longer cardiopulmonary bypass time should be avoided.

Objective EuroSCORE is a widely used objective risk scoring model.The aim of this study was to evaluate the validation of EuroSCORE Ⅱ in predicting mid-term survival after total aortic arch replacement with stented elephant trunk implantation(sun' s procedure) for Stanford Type A aortic dissection.Methods Total 90 patients entered the study randomly.All the patients underwent aortic surgery using total aortic arch replacement with stented elephant trunk implantation (Sun's procedure).The patients were divided into three groups based on the predicting mortality by EuroSCORE Ⅱ.Ggroup 1:0 ＜:P ≤ 5 ％,Group 2:5 ％ ＜ P ≤ 10％,Group 3:P ＞ 10％.Kaplan-Meier method was used to evaluate the long term survival of three groups.Receiver operating characteristic curve was used to test discrimination of the EuroSCORE.Calibration was assessed with a Hosmer-Lemeshow goodness-offit statistic.Results 87 patients were followed umil October,2013.The mean follow-up time was(33.32 ± 11.11) months.Total 5 patients died during the follow-up time.Three patients died in group 1,2 patients died in group 2 and 1 in group 3.There was no statistical difference for the mid-term survival rate between 3 groups (P =0.054).Conclusion Although EuroSCORE Ⅱ is the newest risk model for cardiac surgery,it is not accurate when it is applied for predicting mid-term survival after aortic surgery.A new risk evaluating system specially designed for aortic surgery should be developed in the future.

Objective To investigate the treatment of septic shock(SS) from upper urinary obstruction(UUO). Methods Continuous veno-venous haemofiltration(CVVH) combined with surgical method was applied to 42 SS patients from UUO. Their general conditions, liver and kidney functions, APACHE Ⅱ, therapeutic intervention scorisystem(TISS), multiple organ dysfunction syndrome (MODS), complication rate and main outcomes were analysed comparing with traditional therapies groups(n=30). Results Compared with those traditional therapies, the APACHEⅡ , MODS and TISS score decreased (P＜0.05). Thirty-seven out of 42 patients survived and the survival rate was 88.0% during ICU. Conclusion CVVH combined with surgical methods may effectively decrease the incidence of complications and mortality of SS from UUO, and the mechanism may be related to the remove of mediators of inflammation.

Objective The aim of this study was to evaluate whether the reoperation is the risk factor for mortality after Sun's procedure(Total aortic arch replacement + frozen elephant trunk) for Stanford type A aortic dissection involving aortic arch.Methods Between February 2009 to February 2012,data from 383 patients who underwent Sun's procedure for Stanford type A aortic dissection involving aortic arch were collected retrospectively.35 patients had history of cardiac surgery.Of these patients,16 patients had underwent Bentall procedure,7 patients ascending aortic replacement,4 patients Wheat surgery,4 patients aortic valve replacement,2 patents Bentall combined with mitral valve replacement or plasty,1 patient bivalve replacement,1 patient atrial septal defect repair,1 patient coronary artery surgery.All the risk factors related to mortality were analyzed by univariate statistical analysis.Significant univariate variables were entered into multiple logistic analysis.Results Total 31 patients died in the hospital and the mortality was 8.07％.Of the 35 patients with history of cardiac surgery,3 patients died and the mortality was 8.33％.Univariate analysis showed that symptom onset before surgery less than 1 week,preoperative limb ischemia,combining with coronary artery surgery and cardiopulmonary bypass time longer than 300 minutes in the operative were risk factors for mortality.After these factors were entered into multiple logistic regression analysis,the result showed that symptom onset before surgery less than 1 week (P =0.038,OR =2.43),cardiopulmonary bypass time longer than 300 minutes(PP ＜0.001,OR =12.05) were final independent risk factors for mortality.Reoperation was not the independent risk factor for mortality.The intensive care unit and mechanical ventilation length of reoperation group was (2.09 ± 1.89) days and(30.09 ±33.42) hours respectively,while that of primary group was(2.71 ±3.01) days (P =0.25) and(33.86 ±40.98) hours(P =0.61) respectively.The incidence of postoperative bleeding of reoperation group was 3.03％,while that of primary group was 1.88％ (P =0.50).Conclusion Reoperation was not the independent risk factor for mortality after Sun's procedure for Stanford type A aortic dissection involving aortic arch and the morbidity was also not higher than primary surgery.For these patients,sun' s procedure should be advocated.

Background Retinal microvascular pericytes (RMPs) have been played increasing attention as an emerging key in pathogenesis of various retinal angiogenic diseases including diabetic retinopathy,and RMPs are thought to be a potential target for treatment.Yet the study has been hindered by the difficulty of obtaining source of tissue and isolating pure population.Objective This study was to establish a simple method of isolation,purification and cultivation of primary RMPs for rat.Methods Eyeballs were extracted from clean male Sprague Dawley rats and immersed by 75％ alcohol for 1 minute.The retinas were isolated and mechanical morcel.Trypsin (2.5 g/L) was firstly used and followed by type Ⅰ collagenase (2 g/L) for the digestion of the retina for 15 minutes,respectively.Retinal microvascular fragments were screened by 100 μm and 55 μm filter screen.DMEM containing 20％ fetal bovine serum was added for the cultivation and passaged of the cells.The cells were purified by exchanging medium and partial enzymatic digestion.The morphology and growth status were monitored under the phase contrast microscope,and α-smooth muscle actin (α-SMA),platelet-derived growth factor receptor-β (PDGFR-β),yon Willebrand factor (vWF),glial fibrillary acidic protein (GFAP) antibodies were used for the identification of RMPs.Results RMPs migrated out of fragments after 24-48 hours of plating.On day 7,RMPs appeared in primary cultures as loose colonies.The cells reached confluence to about 80％-90％ on day 14-16.The subcultures grew faster than the primary and reached confluence on day 12-14.The culture showed typical morphology of pericyte with large irregular triangular cell body and multiple long processes,and they could be repeatedly passaged 9 times without obvious loss of characteristic phenotype.Fluorescence assay exhibited that 96％ of the cells showed positive immunofluorescence for α-SMA and PDGFR-β,confirming the purity of RMPs in culture.However,only a few of them were positive for GFAP and the cells response for vWF was absent.Conclusions High purity of rat RMPs can be obtained easily by our method without high cost-consuming.Hcrc wc cstablished a simple mcthod for the primary culture of rat RMPs.

Objective To evaluate the effects of Ang Ⅱ on the expression of IQ domain GTPase-activating protein1 (IQGAP1) and apoptosis of glomerular cells,and to explore the role of IQGAP1 in Ang Ⅱ-induced apoptosis of glomerular cells.Methods Thirty-six male Wistar rats were randomly assigned to receive either saline or Ang Ⅱ by osmotic mini-pump,or be used as normal control.The systolic blood pressure and proteinuria were measured at day 7,14,21,28.After the animals were sacrificed at day 14,28 respectively,the kidneys were collected.Renal pathological change,glomerular cell apoptosis were observed.The expression of glomerular IQGAP1 was assessed by immunohistochemistry,immunofluorescence and Western blotting.The activation of caspase-3 and phosphorylation of extracellular regulated protein kinases 1 and 2 (ERK1/2) were determined by Western blotting.Results Ang Ⅱ-infused rats developed significant hypertension and marked proteinuria.Mild glomerular mesangial cell proliferation and mesangial matrix increase were also observed in Ang Ⅱ-infused rats.The number of apoptotic glomerular cells in Ang Ⅱ-infused rats was significantly more than that in normal control (P ＜ 0.05).The expression of IQGAP1 in glomeruli distributed linearly along the capillary loops.Ang Ⅱ infusion up-regulated the expression of glomerular IQGAP1,which had an significantly positive correlation with activation of caspase-3(r =0.689,P ＜ 0.05) and phosphorylation of ERK1/2 (r =0.658,P ＜ 0.05).Conclusion The enhanced expression of IQGAP1 may be involved in Ang Ⅱ-induced glomerular cells apoptosis via activation of ERK1/2 signaling pathway.

ObjectiveTo investigate the clinical features,therapeutic approach and prognosis of solitary necrotic nodule of the liver (SNN).MethodsClinical data of 10 SNN patients confirmed by operative pathology were analyzed retrospectively.ResultsSolitary necrotic nodule of the liver was found mainly in males (70.0％,7/10),with a median age of 47 years,most of the patients (80.0％,8/10) have no significant clinical symptoms,no underlying hepatitis.Tumor markers including CA19-9,a-fetoprotein and CEA were within normal range.Nine patients had solitary nodule and 1 patient had multiple nodules.Seven lesions located in left lobe and 4 lesions in right lobe.The diameter of the nodules was 2.9 cm ± 1.1 cm.The lesions were low or iso-intense in T1-weighted MR image,low or slight high intense in T2-weighted MR image.Contrast-enhanced MR imaging showed that internal part of lesions had no enhancement,while the capsule of the lesions demonstrated enhancement.MRI image had high diagnostic accuracy rate of 66.7％.A preoperative liver biopsy was difficult to achieve a definite diagnosis.On microscopic examination,all nodules had a central necrotic core,surrounded by fibrotic cells,inflammatory cells and multinucleated giant cells.Furthermore,among them 5 patients have fatty degeneration of the liver.All patients underwent liver resection with no recurrence within 6 - 67 months of follow up.ConclusionsThe etiology of solitary necrotic nodule is unclear.Misdiagnosis was common due to the low incidence and insufficient recognition among physicians,if a definite diagnosis can be obtained,clinical observation is recommended,and the prognosis is good.

Objective To measure the in vitro antibacterial activity of tigecycline against carbapenems-resistant Acinetobacter calcoacetcus-Acinetobacter baumannii complex.Methods The isolated strains of carbapenems-resistant Acinetobacter calcoacetcus-Acinetobacter baumannii complex were collected in our hospital from December 2013 to February 2014.The MIC test strip was a-dopted to measure the MIC value of tigecycline.The break point adopted the judgment criteria published by FDA.Results All 61 strains of carbapenems-resistant Acinetobacter calcoacetcus-Acinetobacter baumannii complex had extremely high drug resistant rate to the commonly used antimicrobial agents.The sensitive rate of tigecycline was 80.3%,intermediation was 19.7% and no re-sistant strain was found in this study.MIC50 and MIC90 were 2 μg/mL and 3 μg/mL respectively.Conclusion Tigecycline has bet-ter in vitro antibacterial activity to the carbapenems-resistant Acinetobacter calcoacetcus-Acinetobacter baumannii complex isolated in our hospital.

Objective To investigate the clinical features,therapeutic approach and prognosis of focal nodular hyperplasia (FNH) of the liver. Methods Clinical data of 34 FNH patients confirmed by operation were analyzed retrospectively. Results Focal nodular hyperplasia of the liver was found mainly in males (67. 6% ) , Mean nodular size was significantly smaller in men than women. Most of the patients (64.7%) had no significant clinical symptoms. Tumor markers including CA19-9 and A-fetoprotein were within normal range. MRI is helpful to achieve a more accurate diagnosis of the disease, correct preoperative diagnosis rate was 54. 5%. All patients underwent liver resection with no recurrence within 17 - 134 months of follow up. Conclusions FHN of the liver is benign in entity. If a definite diagnosis can be obtained preoperatively, observation follow-up are recommended. Hepatectomy is indicated when there is a suspicion of malignancy.