BACKGROUND:The molecule mechanism underlying endothelial progenitor cells(EPCs) differentiation into mature endothelial cells remains poorly understood.It requires for the regulation of the expression of various genes inside cells by various signal pathways,PI3K/Akt maybe included.OBJECTIVE:To investigate the role of PI3K/Akt in EPCs differentiation.DESIGN,TIME AND SETTING:A group control in vitro experiment was performed at the Pathology Laboratory in Shengjing Hospital of China Medical University from September 2007 to March 2008.MATERIALS:Wistar rats aged 4-6 weeks,of either gender.METHODS:EPCs were isolated from rat bone marrow with density gradient centrifugation.Then difference-speed adherence screening method was used to obtain the second time adherent cells,which then were inoculated in culture flask.After 5 days of culture,cells were collected to be indentified with laser confocal microscopy.The differentiating EPCs were those that have positive results of both AC133 and vWF stainings.MAIN OUTCOME MEASURES:The mRNA and the protein expression of AC133,vWF,PI3K and Akt in EPCs were detected with Western blot and reverse transcription-polymerase chain reaction(RT-PCR) after 0,3,7,10 and 14 days of inoculation respectively.RESULTS:According to the RT-PCR and Western blot detection,AC133 showed the strongest expression at day 0,the weaker at day 3 and none at day 7,10 and 14(P

Objective To research the risk factors of new-onset atrial fibrillation (AF) during hospitalization in patients with acute myocardial infarction.Methods Five hundred and one patients first diagnosed with acute ST-segment elevation myocardial infarction who accepted primary coronary intervention from January 2012 to May 2014 in the Second Hospital of Tianjin Medical University were selected.These patients were divided into non-AF group and new-onset AF group depending on whether happened new-onset AF during hospitalization.Data of the patients in acute myocardial infarction complicated with new-onset atrial fibrillation were collected.Univariate and multivariate logistic regression analysis were used to determine the predictors of new-onset AF.Results AF occurred in 49 cases of the 501 (9.8％) patients diagnosed as acute myocardial infarction.Logistic regression analysis showed that admission heart rate(OR =1.019,95％CI:1.004-1.034,P =0.012),left atrial diameter (OR =1.086,95％ CI:1.021-1.156,P =0.009),previous stroke (OR =2.060,95％ CI:1.029-4.123,P =0.041) were independent risk factors for new-onset AF in acute myocardial infarction.Use of β-blockers before coronary intervention treatment can reduce the risk of new-onset atrial fibrillation(OR=0.222,95％CI:0.064-0.768,P=0.017).Conclusion Admission heart rate and left atrial diameter are independent predictors of new-onset atrial fibrillation,and β-blockers in early stage can effectively reduce the incidence of atrial fibrillation.

Objective To investigate the predictive value of CHADS2 and CHA2DS2-VASc score in left atrial (LA) or left atrial appendage (LAA) thrombus in patients with non-valvular atrial fibrillation (AF). Methods A total of 164 consecu?tive non-valvular AF patients confirmed by transesophageal echocardiography (TEE) were included in this study, and were divided into two groups, LA/LAA thrombus group and non LA/LAA thrombus group. The previous history was recorded in two groups of patients. Their CHADS2 and CHA2DS2-VASC scores were calculated based on the general data of two groups. The laboratory examinations and TEE indexes were compared between two gropes. Results Of all patients, the rate of LA/LAA thrombus was 19.5%. The CHADS2 and CHA2DS2-VASC scores were significantly higher in LA/LAA thrombus group than those of non LA/LAA thrombus group (2.1±1.3 vs 1.0±0.9, 3.4±1.8 vs 1.9±1.4,P<0.01). There were 13(11.9%) and 5 (8.8%) patients showed LA/LAA thrombus under low-moderate risk in the two score systems, respectively. Multivariate logis?tic regression analysis showed that CHADS2 score≥2 points(OR=3.735, 95%CI:1.508-9.251, P=0.004) and CHA2DS2-VASc score ≥2 points (OR=5.104, 95%CI:1.586-16.425, P=0.006) were independent risk factors of LA/LAA thrombus. ROC curve showed that AUC of CHADS2 and CHA2DS2-VASc scores to predict LA/LAA thrombus were 0.731 (95%CI:0.630-0.832, P<0.001) and 0.742 (95%CI:0.640-0.843, P<0.001), respectively. Conclusion CHADS2 and CHA2DS2-VASC score ≥2 points are independent risk factors of LA/LAA thrombus in patients with non-valvular atrial fibrillation. The pre?dictive values of both score systems are moderate. There are also LA/LAA thrombus in low-moderate risk patients when us?ing the two score systems, and with the increased risk stratification, the rate of LA/LAA thrombus is increased.

OBJECTIVE To establish the basic quality control system for disinfection of medical apparatus and(instruments).in order to meet the national standards of the disinfection quality.METHODS On the basis of background investigation about the disinfection(management) of medical apparatus and instruments from 1997 to 2004 to implemeat the basic quality control system scientifically and continuously and to improve the management of disinfection quality in hospital.RESULTS The disinfection quality of commonly used medical apparatus and(instruments),first-aid devices,combat readiness materials,special instruments and sterilizer,etc was got to(improve) year by year,that created condition to control the emergence of hospital(infection).CONCLUSIONS To(avoid) the exogenous hospital infection due to unsuitable medical apparatus and(instruments).

Objective To explore the relationship between the expression of Ki67 and mitosis karyorrhexis index（MKI）in ganglioneuroblastoma（GNB）.Methods Clinical materials and histology slides from 28 patients with GNB were reviewed.MKI was counted under microscope and the expression of Ki67 protein in GNB was evaluated by immuohistochemical technique.The follow-up data were also collected and statistically analyzed.Results There were 15 patients with favourite prognosis（favourite subgroup,FS） and 13 patients with poor prognosis（unfavourite subgroup,US）.The survival time of GNB patient in FS was significantly longer than that in US（P 0.05）.The survival time is related to pathology classification（P 0.05）.MKI was correlated with advanced clinical stage,pathology classification and survival time（P 0.001）.No significant difference in Ki67 expression was found between low and high MKI patients.Conclusion MKI might be a reliable prognostic parameter for making diagnosis and evaluating prognosis of GNB.Detection of Ki67 expression might be not useful for evaluating the prognosis of GNB.

Objective To investigate the nephroprotective effects of prostaglandin E1 (PGE1) in the elderly undergoing coronary angiography or intervention. Methods161 patients undergoing coronary angiography (CAG) or percutaneous coronary intervention (PCI) were randomly assigned to PGE1 group (n=87 cases) and control group (n=74 cases).10 μg lipo-PGE1 added to 100 ml normal saline were administered intravenously once daily for 5 days before and 2 days after the operation.The serum levels of creatinine (Scr) and cystatin C (Cys C) were measured on admission and 48 h after the procedure.Results After the procedure,the levels of Scr and Cys C were increased (P＜0.01) and creatinine clearance (Ccr) was decreased (P＜0.05) in control group than in PGE1 group.The incidence of contrast-induced acute kidney injury (CI-AKI) in control group 〔12.2％ (9/74)〕 was higher than in PGE1 group 〔3.4％(3/87)〕 (P＜0.05).The application of P(GE1 decreased CI-AKI,but high basic level of Scr and diabetes mellitus enhanced the incidence of CI-AKI by logistic regression.The serum levels of Cys C had negative correlation with Ccr (r=-0.615,P＜0.01).Conclusions Perioperative application of PGE1 has nephroprotective effects in the elderly undergoing CAG or PCI,and decreases the incidence of CI-AKI.The serum levels of Cys C is one of ideal indexes for auxiliary diagnosis of CI-AKI.

Objective To investigate the expression and relationship of TIP30(HIV-1 Tat interactive protein 2), vascular endothelial growth factor (VEGF) and MVD (detected by CD34) in the angiogenesis of human brain astrocytomas. Methods Expression of TIP30, VEGF and CD34 in 19 cases of normal brain tissue and 71 cases of astrocytoma were immunohistochemically examined with Elivision plus two-step method. Results The positive expression of TIP30 could be seen in cytoplasm of neuroglial cells and neurons of 19 normal brain tissues. The positive expression rate of TIP30 in 71 cases of astrocytoma was 33.80 % (24/ 71). The positive expression rate of TIP30 in astrocytoma of different grades was 52 % for grade Ⅱ, 34.78 % for grade Ⅲ and 13.04 % for grade Ⅳ. The positive expression rate of TIP30 in high grade (Ⅲ+Ⅳ) of astrocytoma was found significantly lower than that in low grade(Ⅱ) (χ2=5.71, P <0.05); The expression of VEGF and MVD detected by CD34 in astrocytomas were higher than that in normal brain tissue and increased as the tumor grade increased; In astrocytoma, the negtive correlation was found between the expression of TIP30 and VEGF (r=-0.428, P<0.05); no correlation was found between TIP30 and MVD(r=-0.065, P 0.05); the positive correlation was found between VEGF and MVD(r=0.684, P<0.01). Conclusion The positive expression rate of TIP30 in normal brain tissue is significantly higher than that in astrocytoma. The positive expression rate of TIP30 significantly decreases as the pathological grade of the astrocytoma increases; The expression of TIP30 and VEGF is negatively correlated in astrocytoma.

Objective To investigate the efficacy and pregnancy outcomes of women receiving double-catheter epidural block in labor analgesia, and compare the results with single-catheter epidural block.Methods A double-blind clinical trial was conducted on 206 full-term singleton primiparas, aged 25-35 and at the 37 -42 weeks of gestation who delivered at the Department of Obstetrics, Qingdao Municipal Hospital from August 2006 to December 2008, which were randomly divided into two groups:double-catheter epidural block ( group D, n = 103) and single-catheter epidural-block ( group S, n = 103 ).Women in group D were given mixture of 0.1% repivacaine hydrochloride and 0.5 mg/L sufentinil 4 -6 ml as initial dose.Patient control epidural analgesia pump (PCEA) was connected with the upper catheter after 45 minutes.A bolus dose of 4 -6 ml analgesia mixture was infused according to the condition through the lower catheter.Women in group S received analgesia mixture 10 - 15 ml as initial dose and PCEA pump was connected after 45 minutes.Oxytocin was infused in both groups according to uterine contraction after 30 minutes.The following indexes was observed: ( 1 ) visual analogue scales (VAS); (2) modified Bromage Scores;(3) the total dose of analgesia mixture, the percentage of oxytocin infusion, duration of labor and duration of the second stage of labor; (4) fetal birth weight and Apgar scores( 1,5 minutes); (5) mode of delivery; (6) the concentration of plasma cortisol and angiotension Ⅱ at the beginning of regular uterine contraction and at the time when cervical dilated to 4 cm and 10 cm and fetal disengagement; (7)anesthesia-related complications.Results ( 1 )The neonatal birth weight and Apgar scores ( 1,5 minutes)of group D were (3456 ±468)g, 9.8 ±0.6 and 9.9 ±0.7, respectively, while(3399 ±569) g, 9.8 ±0.5 and 9.9 ±0.7 in group S( P ＞0.05).No motor function block was reported in any group and the modified Bromage score was zero.(2) The total dose of analgesia mixture in group D was similar to that in group S [(57 ±9) ml vs.(58 ±11) ml, P＞0.05].However, the percentage of women received oxytocin in group D was smaller [59.2% (61/103) vs.81.6% (84/103), P ＜ 0.01], and the total time of labor and the duration of second stage of labor in group D were shorter[(532 ± 140) minutes vs.(608 ± 150) minutes;(46 ± 31 ) minutes vs.(60 ± 34) minutes, P ＜ 0.05].(3) There were no significant differences in VAS at 30 minutes after initial dose and in the first stage of labor between group D and S ( 1.2 ± 1.1 vs 1.2 ± 1.1,1.1 ± 1.1 vs.1.2 ± 1.0, P＞0.05).VAS at the second stage of labor stage was lower in group D than in group S ( 1.2 ± 1.1 vs.4.5 ± 2.2, P ＜ 0.01 ).(4) The rate of cesarean section, instrumental delivery and episiotomy in group D were lower than in group S (7.8% vs.17.5%, 7.8% vs.15.5%, 10.7% vs.18.4%, P ＜ 0.05).The incidence of fetal distress and meconium-stained amniotic fluid as the indication of cesarean section were similar between the two groups (P ＞ 0.05 ).Lower incidence of fetal malpresentation and arrested second stage of labor were shown in group D than in group S (2.9% vs.9.7%, 1.0% vs.5.8%, P ＜ 0.05 ).(5) The concentration of plasma cortisol and angiotension Ⅱ were lower in group D than in group S [(86 ±25) ng/L vs.( 100 ±20) ng/L, (278 ±53) nmol/L vs.(311 ±53)nmol/L, P＜0.05] only at the end of second stage of labor, but not at any other times(P ＞0.05).(6) No serious anesthesia-related complications were reported in any groups.Some light backache around the puncture point were complained by 29.1% (30/103) of the women in group D and 31.1% (32/103) in group S(P ＞0.05).Conclusion Double-catheter epidural block can provide better analgesia effect during labor than single-catheter epidural block, without any adverse influence on delivery outcomes.

OBJECTIVETo investigate the anti-human CEM lymphoma cell activities induced by TCR idiotypic DNA vaccine containing different antigen determinants in BALB/c mice.

METHODSThe specific rearranged gene fragment encoding TCRVbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into eukaryocytic expression vector pcDNA3, which was used as DNA vaccine and template for PCR amplifying different antigen determinant. Gene fragments encoding different antigen determinant were amplified and cloned into pcDNA3, separately. The experimental mice were immunized by intramuscular injection of the DNA vaccines. The specific anti-idiotype antibodies were detected by indirect immunofluorescence assay.

RESULTSTCRbetaV of CEM cell line contains five antigen determinants. Specific anti-idiotype antibody was detected in all of the six mice immunized with DNA vaccine containing all the five determinants (the highest titer was 1:480). Although the antibody could also be detected in four of the six mice immunized with DNA vaccine containing four of the five antigen determinants, the antibody titer was lower (the highest titer was 1:80). DNA vaccine containing two of the five determinants could not induce the specific antibody.

CONCLUSIONThe idiotypic DNA vaccine containing the whole TCRbetaV five antigen determinants could induce the specific anti-lymphoma idiotypic antibody in BALB/c mice.

Amino Acid Sequence ; Animals ; Antibodies, Anti-Idiotypic ; blood ; immunology ; Base Sequence ; Complementarity Determining Regions ; genetics ; immunology ; Epitopes ; genetics ; immunology ; HL-60 Cells ; Humans ; Lymphoma ; immunology ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; immunology ; Sequence Analysis, DNA ; Tumor Cells, Cultured ; Vaccines, DNA ; genetics ; immunology

This study was undertaken to investigate the anti-lymphocytic malignacy immunologic effects induced by TCR idiotypic DNA vaccine on BALB/c mice. CEM lymphoma cell line and BALB/c mice were used as models. The rearrangement gene fragment coding TCR Vbeta region of CEM cell line was obtained by RT-PCR technique. The PCR product was cloned into the eukaryocytic expression vector pcDNA3 to be used as DNA vaccine. The experimental animals were immunized by intramuscular injection with DNA vaccine. The specific anti-idiotypic antibody was detected by indirect immunofluorescence assay. The specific anti-idiotypic cellular immunity was detected by CTL activity assay using MTT method. The results showed that specific anti-idiotypic antibody in the immunized mice sera could be found since four weeks after immunization and came to the peak of titer on the sixth week. Using IL-6 as immunological adjuvant could significantly increase the antibody titers. It was concluded that the TCR idiotypic DNA vaccine could induce effectively the specific anti-lymphoma idiotypic antibody in BALB/c mice. Using IL-6 as immunological adjuvant could significantly increase the antibody titers induced by idiotipic DNA vaccine.