BACKGROUND: The human leucocyte antigen (HLA) allele are now understood to be alternative DNA sequences at the same physical gene locus, which may or may not result in different phenotypic traits. The generation of a new allele is induced by various factors, but, whether the mutation would be exist after removing the causative factors need further investigation. OBJECTIVE: To confirm the new allele by DNA sequencing technique, in addition, to analyze the heritage of a HLA allele A~*9217 (No. of WHO registration: WHS10004629)DESIGN, TIME AND SETTING: The open experiment with DNA as observation object. The initial detection with polymerase chain reaction sequence-specific oligonucleotide (PCR-SSO) was performed at HLA Laboratory of Henan Provincial Red Cross Blood Center in November 2007, and the sequencing was performed at HLA Laboratory of DYNAL Biological Technology (Beijing) Co., Ltd. in February, 2008. MATERIALS: The proband (sample ID: 371xxxxx ) and other 8 family member was investigated, blood sample was collected at the HLA Laboratory.METHODS: Nine family members of A~*9217 carrier were typing for HLA- A, B, DRB1 using PCR-SSO and SBT methods for low-media and high resolution, and 3 red blood cell blood groups systems: ABO, MN and Rh was tested to assist analysis .MAIN OUTCOME MEASURES: Sequence alignment of HLA-A exon 3 in A~*9217 carriers.RESULTS: According to the results of the blood groups phonotype of ABO, MN, Rh systems and HLA, the new allele A~*9217 of the proband was paternal origin. However, the sequence of this allele is A*020301, which had 3 base difference in exon 3, the new allele appears 3 base change at 391 T>G, 414 C>G, 418 T>G, and this new sequence was found in proband's two children.CONCLUSION: The new allele A~*9217 is generated from proband's father mutation, which can pass it to his children. However, the further heritage of A~*9217 allele need exploring.

AIM To investigate the positive inotropic effect of aconitine on isolated hearts from cardiac dysfunctional rats when combined with pinacidil. METHODS In our experiment Langendorff perfusion equipment was used to investigate the following cardiac indexes:LVSP(left ventricular systolic pressure), LVDP(left ventricular diastolic pressure), +d p /d t max (the maximum going up rate of left ventricular pressure) and -d p /d t max (the maximum going down rate of left ventricular pressure) from rat hearts under two conditions:① aconitine only,② aconitine plus pinacidil(an agonist of K ATP channel) to observe the positive inotropic effect of aconitine on cardiac dysfunctional rat hearts. RESULS ①Aconitine had certain positive inotropic effect on isolated hearts from cardiac dysfunctional rats; ②Combing aconitine with potassium channel activator markedly winded the doses range between the effective and toxic concentration and enhanced the cardiotonic effect of aconitine. CONCLUSION Aconitine had positive inotropic effect on dysfunctional isolated rat hearts; the combination of aconitine with potassium channel activator markedly widened the cardiotonic doses range of aconitine and enhanced its cardiotonic effect.

Mutations and deletions of the tumour suppressor PTEN are frequently involved in the development of cancer.However, PTEN is also tightly controlled by various non-genomic mechanisms,such as the epigenetic silen-cing of PTEN, post-transcriptional regulation by non-coding RNAs and post-translational modification.

Objective To investigate and compare the effects of minimally invasive percutaneous nephrolithotomy for renal upper calyceal stones with approaches through upper edge of the 1 lth rib and lower edge of the 11th fib.Methods Two hundred and sixteen cases of kidney stones in our hospital from July 2008 to July 2010 were reviewed.Of which 66 patients were renal calyceal stones.Thirty-two patients underwent the approach through upper edge of the 11th rib (group A),with 15 males and 17 females,with a mean age of 42 years.Ten cases with stone were located on the left side,right 21 cases,and bilateral 1 case,with calculi average maximum diameter of 1.9 cm.Thirty-four cases underwent the approach through lower edge of the 11th path (B group),with 14 males and 20 females,with a mean age of 44 years.Thirteen cases with stone were located on the left side,right 21 cases,with calculi average maximum diameter of 1.7 cm.The two groups underwent minimally invasive percutaneous nephrolithotomy holmium laser lithotripsy lithotomy.The average puncture time,one-time success rate of puncture and disposable stone-free rate were compared.Results Puncture time of group A was 3.0-14.0 min,with an average of 5.8 min,and group B was 3.0-17.0 min with an average of 6.4 min.The disposable puncture success rate of the two groups was 100％.Disposable stone-free rate was 100％ in group A (32/32),and 88％ in group B (30/34) (P ＜ 0.05).There were no pneumothorax,hemothorax,abdominal viscera injury in two groups.Conclusions The minimally invasive percutaneous nephrolithotomy through the approach of upper edge of the llth fib was better than through the approach of conventional lower edge of 11 subcostal margin,with shorter percutaneous renal access distance,and higher stone-free rate.

Objective To investigate the plasma level of D-dimer in healthy population and hospitalized patients, and to analyze the cut-off value of D-dimer levels in different age groups for the exclusion of venous thromboembolism (VTE) in older patients. Methods A total of 6 717 healthy individuals and 2 976 hospitalized patients were included in the study. The blood level of D-dimer was detected by the Biomerieux VIDAS fluor-euzymelinked immunoassay analyzer assay. Results The blood level of D-dimer was significantly increased with age in healthy people aged over 40 and different age groups of hospitalized patients (P<0.01). The incidence of venous thromboembolism was also increased with age. The cut-off value for the diagnosis of VTE with D-dimer was 504 μg/L in patient group aged ≤40 years, and the sensitivity was 100.0%. The cut-off value was 565μg/L in 41~60-year group, and the sensitivity was 96.2%. The cut-off value was 817μg/L in 61~74-year group, and the sensitivity was 80.8%. The cut-off value was 1 024μg/L in≥75-year group, and the sensitivity was 73.1%. Conclusion The plasma D-dimer level increased with age. The D-dimer cur-off value was higher in older patients than that of other age groups, but the sensitivity in diagnosis was decreased obviously.

Aim This paper aims at investigating the effect of dofetilide on Na+/Ca 2+exchange in ventricular myocytes of adult guinea pigs. Methods Rapid enzymatic isolation procedure was performed to get single ventricular myocytes from adult guinea pigs. After the whole cell configuration was formed, a ramp voltage-clamp pulse depolarized from a holding potential of -40 mV to +60 mV, then hyperpolarized to -120 mV at a rate of 90 mV/s to record the Na+/Ca 2+ exchange current (I Na/Ca) and the influence of dofetilide from 0.03～1.0 ?mol?L -1. Result Dofetilide from 0.03～1.0 ?mol?L -1 concentration-dependently increased the I Na/Ca in both outward and inward mode. The EC 50 of dofetilide on I Na/Ca in outward and inward mode of NCE was 0.178 ?mol?L -1 (95% confidence interval was 0.040～0.787 ?mol?L -1) and 0.178 ?mol?L -1 (95% confidence interval was 0.038～0.842 ?mol?L -1), respectively. Conclusion Dofetilide concentration-dependently increased the outward and inward Na+/Ca 2+ exchange current in ventricular myocytes of adult guinea pigs.

Aim To investigate the effects of 5-HT_4 receptor agonist and 5-HT_3 receptor antagonist 2-[1-(4-piperonyl)piperazinyl]benzothiazole on rat heart rhythm and the involved ionic mechanisms.Methods Langendorff-perfused rat hearts were subjected to 0.1～10 μmol·L~(-1) 2-[1-(4-piperonyl)-piperazinyl]benzothiazole for 15 minutes with simultaneous ECGs recording.The whole-cell patch-clamp electrophysiology was used to record effects of 2-[1-(4-piperonyl)piperazinyl]benzothiazole on inward rectifier K~+ current(I_(K1)),transient outward K~+ current(I_(to)),resting membrane potential(RMP)and action potential(AP)in enzymatic dissociated rat ventricular myocytes.Results In ex vivo Langendorff-perfused hearts,0.1～10 μmol·L~(-1) 2-[1-(4-piperonyl)piperazinyl]benzothiazole elicited singnificant rhythm disturbances.In the presence of 10 μmol·L~(-1) agent,the total of PVB were 236±37,87.5%(7/8)hearts exhibited VT,and 62.5%(5/8)hearts exhibited VF(P<0.01).At the concentration of 0.1～10 μmol·L~(-1),2-[1-(4-piperonyl)piperazinyl]benzothiazole could inhibit I_(K1)(EC50=0.74 μmol·L~(-1))and I_(to)(EC50=2.16 μmol·L~(-1)),decrease RMP and prolong action potential duration(APD)in concentration-dependent manners(n=6,P<0.01).Conclusion Inhibition of IK1,Ito and resultant prolongation of APD,depolarization of RMP might be the critical causes for induction of arrhythmias by 2-[1-(4-piperonyl)piperazinyl]benzothiazole in rat.

Aim To assess the effects of N -[2-p-bromo-cinnamylamino-ethyl]-5-isoquinoline-sulfonamide (H-89), a potentially selective inhibitor of Protein Kinase A (PKA), on cardiac membrane ion channels and transporters, which will further fulfill our understanding of pharmacology of PKA inhibitors.Methods Whole-cell patch clamp was used to investigate the effects of H-89 on cardiac L-type Ca~(2+) current (I_(Ca-L)), Na~+ current (I_(Na)), inward rectifier K~+ current (I_(K1)), transient outward K~+ current (I_(to)) and Na~+-Ca~2+ exchanger current (I_(Na/Ca)) in enzymatic dissociated SD rat ventricular myocytes.Results H-89 at 1～10 μmol·L~(-1) could inhibit I_(Ca-L) , I_(Na) , and Ito in a concentration-relative manner (P <0.05). At low concentra-tion (5 μmol·L~(-1)), H-89 completely inhibited I_(K1) (P <0.05) just as the action of 0.5 mmol·L~(-1) BaCl_2.Further, H-89 at 1～10 μmol·L~(-1) had no significant effect on I_(Na/Ca) (P >0.05).Conclusion The direct or PKA-mediated indirect action maybe involved in the effects of H-89 on ion currents and transporter.

Objective To investigate the reasonable time limit for stopping the aspirin treatment in preoperative pa-tients with general surgery and the effects on platelet function in postoperative patients with recovering the therapy of aspirin. Methods A total of 121 patients undergoing elective general surgery were divided into stopping aspirin treatment 5 d group (n=59) and stopping aspirin treatment 7 d group (n=62). Fifty healthy volunteers were used as the control group. The arachi-donic acid (AA)-induced platelet aggregation test was used to detect the platelet agglutination rate in all groups. Aspirin was reused 24~48 h after surgery. The level of urinary 11-dehydro-thromboxane B2 (11-DH-TXB2) was assayed by ELISA 7 and 10 d after retreatment. Results The levels of the PAgT (5 min, 8 min and 10 min) were decreased significantly in pa-tients with stopping aspirin treatment 5 d group compared with those of patients with stopping aspirin treatment 7 d group and control group (P<0.05). There was no significant difference in the level of PAgT between patients with stopping aspirin treatment 7 d group and control group. The platelet aggregation showed two different characteristic curves after stopping aspi-rin treatment for 5 d. And the normal curve of platelet aggregation was found after stopping aspirin treatment for 7 d. The lev-els of PAgT and urinary 11-DH-TXB2 were significantly lower in patient recovered the aspirin treatment for 7 d and 10 d than that of control, and which was significantly higher in 7 d group than that of 10 d group (P>0.05). Conclusion The platelet aggregative function returned to normal level in patients with 7-d preoperative stopping aspirin. The laboratory moni-toring of aspirin therapy should be more than 7 d after postoperative reusing aspirin.

Objective To study the variation characteristics of plasma von Willebrand factor antigen (vWF Ag) after the off-pump coronary artery bypass graft (OPCAB) and its predictive value on the assessment of cardiovascular ischemia events in the postoperative patients.Methods A total of 338 patients with non-ST-segment elevation myocardial infarction were selected from 2010 to 2012 in this retrospective cohort study,with 249 males and 89 females and a mean age of (69.2 ± 4.5) years.The level of vWF Ag was assayed by the IL ACL-TOP 700 blood coagulation instrument.Receiver-operator curve (ROC) analysis of vWF Ag levels in the prediction of risk of ischemic events was performed.x2 test and Logistic regression were conducted to analyze the relevance between vWF Ag and clinical pathological factors.Cox regression analysis model were used to evaluate the effect on prognosis.Results There was significant difference in vWF Ag level at different time point between the poor recovery group and the stable disease group (x2 =129.53 and 101.48 respectively,both P＜0.01).And the vWF Ag level was higher in the poor recovery group on the 14th,30th,60th,90th day after OPCAB than in the stable disease group at the same time points respectively (all P＜0.05).The optimum cut-off point of vWF Ag (the 30th day after OPCAB) for prediction of ischemic events was 251 ％ within the 31-90th day after OPCAB,the area under ROC curve was 0.839 (95％ confidence interval:0.776-0.902).The Logistic regression analysis showed that the vWF Ag level on 30th day after OPCAB was affected by age,left ventricular ejection fraction,left main artery disease,serum creatinine level,vascular number of bypass grafts,history of myocardial infarction and hypertension (all P＜0.01).The Cox analysis showed that the cut-off value of vWF Ag (on the 30th day after OPCAB) was the independent prognostic assessment parameter for ischemic events in patients with cardiovascular diseases within one year.Conclusions The variation of serum vWF Ag level can reflect the integrated influence of multiple pathological factors on vascular endothelial function after the OPCAB,it can become the effective predictor for disease progression within 90 days and the long-term prognosis after OPCAB.