Objective To investigate the alternations in gene/amino acid sequence of penicillin-binding protein (PBP)2x from clinical isolates of Streptococcus pneumoniae, and to find the reasons for the rapid surge of penicillin and cefotaxime nonsusceptibility among pneumococcal isolates from Shenyang. Methods Thirty-four strains of Streptococcus pneumoniae were collected from January 2006 to February 2007. The antibiotics susceptibility of these strains was detected. PCR amplification and direct sequencing of pbp2x genes were performed. The sequence variations of PBP genes of the penicillin nonsusceptible Streptococcus pneumoniae(PNSP) in this region were studied by BLAST analysis. Results Two prominent substitutions were common to 12 PNSP isolates for which the MIC of penicillin resistance and cefotaxime were at least 0.5 mg/L, which included the replacement of Thr338→Ala in the first conservative motif STMK and Leu546→Val adjacent to third conservative motif KSG. The importance of the exchange of His394→Leu was identified in one PNSP isolate 15. The remarkable finding in this study was Met342→Ile following the first conservative motif STMK. pbp2x sequences of eight PRSP isolates shared Lys501-Glu505-Thr507 substitutions which might be served as a unique marker for PRSP in this region. Novel gene and amino acid sequence variants in 17 isolate were identified in this study, and these sequences have been deposited in the GenBank database and assigned accession No. EU044831, EU089706-EU089709, EU106881-EU106884 and EU124672. Conclusion It is likely that the emergence of penicillin and cefotaxime nonsusceptible Streptococcus pneumoniae in Shenyang might be associated with novel gene sequence variants.

Objective To explore the changes of left atrial systolic function in patients with coronary heart disease after coronary artery bypass grafting (CABG). Methods Strain rate imaging (SRI) was performed on 23 patients with coronary heart disease before CABG, 1 week, 1 and 3 months after CABG to evaluate left atrial systolic function quantitatively. Results No significant change of left atrial systolic function was detected 1 week after CABG (P＞0.05 ). E/A and LVEF increased, LAFS, AEF and SRa decreased 1 month after CABG compared with those before CABG (P＜0.05). Three months after CABG, changes turned more significantly (P＜0.01). Left ventricular ejection fraction (LVEF) increased 1 and 3 months after CABG, and its changing rate negatively correlated with those of Sra (r=-0.751,-0.783; all P＜0.01). Conclusion Left atrial systolic function is affected by CABG, presenting as decrease of pump function. SRI can be used to evaluate the atrial systolic function quantitatively and monitor the changing of left atrial systolic function dynamically after CABG.

Objective To observe the growth situation of Blastocystis hominis in vitro and select the optimal method for culti?vation of B. hominis in different media. Methods Ten positive stools with B. hominis were inoculated in three different media for cultivating,namely 1640,Jone’s medium and vitro medium. And the stools with good growth status and high quantities of B. hominis were chosen to inoculate in the three media with equal amount after subcultivation,and the number of B. hominis was counted every 24 h for ten days,and the morphological changes and growth status were also observed. Results The densities of B. hominis in the 1640 and Jone’s medium were higher than that in the vitro medium 48 h after the inoculation. The same stool sample was inoculated to the three different media and observed for ten days,and the results indicated that the growth of B. homi?nis presented regular changes in the three media,the growth peaks were on the third,sixth and ninth day post inoculation;and the density of B. hominis was the highest in the Jone’s medium. The morphology of B. hominis was the clearest and most dynamic in the vitro medium,while various reproductive forms were observed in the Jone’s medium. Conclusion Jone’s medium is suitable for the growth of B. hominis and can be the first choice for the cultivation of B. hominis in vitro,and vitro medium is the best medium for observing the growth situation of B. hominis.

Objective To explore the relations among blood lipid,blood rheology and microcirculation of bulbar conjunctiva of model rats of diabetesⅡwith heart blood stasis syndrome.Methods The rats were fed with high fat and caloric feed.Intraperitoneal injection of streptozotocin(STZ)in small dosage was applied to make the model of diabetesⅡ.Then the model of heart blood stasis syndrome was made through introperitoneal injecting dexamethasone and adrenalin.Eight hours after empty stomach,the rats were anaesthetized to measure the microcireulation of bulbar conjunctiva and test the blood lipid and carotid blood rheology.Results The indices of blood lipid,blood rheology and bulbar eonjunetiva microcirculation of model rats were all changed to abnormal,tallying with the changes of cardiovascular complications of diabetes.

Blood hypercoagulabale state of malignant tumor was the pathological basis of tumor blood stagnation,which led to tumor proliferation and metastasis and also influenced the therapeutic effect and prognosis of tumor.Based on knowledge of thrombin and tissue factor system,We discussed the mechanism blood hypercoagulabale leading to tumor proliferation and metastasis,and the influence of traditional Chinese drugs of activating blood circulation to dissipate blood smsis on tunlor proliferation and metastasis.We concluded that traditional Chinese drugs have sound effects on prevention and treatment of tumor proliferation and metastasis.

Objective To investigate the alternations in gene/amino acid sequence of penicillin-binding protein (PBP) 2b from clinical isolates of penicillin-nonsusceptible Streptococcus pneumonia(PNSP) in this region.Methods 24 strains of Streptococcus pneumonia were collected from January to December 2006.The antibiotics susceptibility of these strains was detected.PCR amplification and direct sequencing of pbp2b genes were performed.The sequence variations of PBP genes of the PNSP in this region were studied with sequence BLAST analysis.Results Three prominent substitutions were common tO 13 PNSP isolates with minimal inhibitory concentration(MIC) at least 0.1 mg/L.These included the replacement of Thr445→Ala following the conservative motif SSN,Glu475→Gly and Thr488→Ala/Ser.The exchange of Glu332→Gly was identified in 12 PNSP isolates of which the MIC was at least 0.25 mg/L.Seven penicillin resistant Streptococcus pneumonia (PRSP) isolates (MIC≥3 mg/L)shared the amino acid substitution Ala618→Gly adiacent to third conserved (KTG) motif and the PBP2b sequences of seven PRSP isolates were classified within Back's group Ⅱ and were very similar to those of the Korean J77 isolate.Novel gene and amino acid sequence variants in isolate 14,15,8,11 and 24 was identified in this study and these gene sequences have been deposited in the GenBank database and assigned accession no.EU035970,EU056919,EU056920,EU056921 and EU106886.Conclusion Analysis of pbp2b genes revealed highly similar patterns of nucleotide and amino acid sequence variation among most resistant isolates.while penicillin intermediate Streptococcus pneumonia might be associated with novel gene sequence variants.

Objective To determine the possible genetic background and the source of our hospital's 43 clinical isolates of multidrug-resistant Acinetobacter baumannii, and the category of gene cassettes in type 1 integrons of all strains.Methods Restriction enzyme Apa I was chosed for all strains in pulsed-field gel electrophoresis (PFGE) methods.Multilocus sequence typing ( MLST) was used to compare the allelic profiles of all the strains. PCR method was used for amplify the integrons of all strains. Results PFGE results showed that 43 strains were divided into four types. A-type and B-type were divided into 4 and 2 subtypes, respectively. The MLST results showed the existing of three allelic profiles; 1-3-3-2-2-7-3, 1-3-3-2-2-11-3, and 1-3-3-2-2-14-3.B-type and D-type of PFGE have the same allelic profile(1-3-3-2-2-11-3).A-type strains were detected mainly in ICU, and in burn unit only found B- and D-type.The same integron was detected in 62.8% of the strains.The constituent ratio of A1,A2,A3,A4,B1,B2,C and D-type was 40.7% , 18.5% , 7.4% , 3.7% , 14.8% , 3.7% , 3.7% and 7.4% , respectively.Conclusions The coexistence of multiple cloning system in this region was proved by the PFGE and MLST, and the same clone can evolve to different subtypes when stimulated by different environmental conditions; and the different carrying-situationt of the same integron in strains prove the possibility of the change during the evolution of resistance mechanisms.

Objective To explore the effect of coronary artery bypass grafting(CABG) on left atrial (LA) function by strain rate imaging(SRI). Methods Twenty-three patients with coronary heart disease who underwent coronary artery bypass grafting were involved. SRI was performed on those patients to evaluate LA function quantitatively at baseline (before CABG),and at 1 week, 1 month and 3 months after CABG. Peak strain rate(SR) was measured at each segment (septal, lateral, posterior, anterior, and inferior walls) and mean peak systolic SR (SRs),peak early diastolic SR (SRe) and peak atrial systolic SR (SRa) were calculated by averaging data in each segment. Results Compared with the baseline,LV pre-systolic volume(LAVp), maximal volume (LAVmax), minimal volume (LAVmin), LV active emptying fraction (LAAEF) and passive empting fraction(LAPEF) had on significant differences at 1 week (P >0.05). LAVp,LAVmin,LAVmax and LAAEF decreased gradually after CABG, LAPEF increased gradually after CABG (P <0.05). Compared with the baseline, the peaks of SR curve showed no significant differences at 1 week (P >0.05). Nevertheless,the peaks of SR were increased at systole and early diastole,decreased at atrial contraction at 1 month (P <0.05). Those changes were turned more significantly at 3 months (P 0.01). Left ventricular ejection fraction (LVEF) both increased at 1 month and 3 months,and its changing rate correlated inversely with the changing rate of SRa respectively (r = -0.751, -0.783,all P<0.01).Conclusions LA function is affected by CABG, presented as reservoir and pump functions decreased and conduit function increased. SRI can evaluate the atrial function quantitatively and monitor the changing of LA function dynamically after CABG.

Objective To detect the encoding gene of efflux pump and two-component system, and investigate the effect of efflux inhibition on the multiresistance of Acinetobacter baumarmii. Methods PCR was used to detect the adeB, adeR and aries gene. Agar dilution was used to determine the minimum inhibitory concentrations(MIC) of ciprofloxacin, cefotaxime, amikacin and imipenem of 50 multiresistance Acinetobacter baumannii, with or without 25 μg/ml reserpine. Results 94%, 96% and 92% of 50 muhiresistance of Acinetobacter baumannii were detected for adeB, adeR and aries gene,respectively. At least four fold decrease of MIC was observed in 49, 50, 50 and 46 isolates for ciprofioxacin, cefotaxime, amikacin and imipenem, respectively. Conclusion The multiresistance of Acinetobacter baumannii is related to the effect of the efflux system.

Objective To exPlore the effect of mammalian target of raPamycin ( mTOR ) inhibitor eVerolimus on radiosensitiVity of human non_small cell lung cancer cell line in vitro by using eVerolimus to inhibit mTOR signaling Pathway of A549. Methods Human non_small cell lung cancer cell line A549 was subjected to radiation alone or in combination with eVerolimus treatment. The 50%inhibition concentration ( IC50 ) of eVerolimus in A549 cells was detected by methylthiazol tetrazolium ( MTT) assay in vitro. EVerolimus at the 20%inhibition concentration ( IC20 ) was used to Pretreat A549 cells for 24 h. Cells were then irradiated by X_ray with 2,4,6,8 Gy. The cell surViVal fraction was comPuted by clone formation. Cell surViVal curVe was fitted by multitarget one_hit model, and mean lethal dose ( D0 ), dose quasithreshold ( Dq ), surViVal fraction at 2 Gy ( SF2 ), and sensitization enhancement ratio (SER) were calculated. The exPression ofγ_H2AX was determined by Western blotting and then the relatiVe gray Values were analyzed. Results EVerolimus significantly imProVed the sensitiVity of A549 cells to radiation. The D0 , Dq and SF2 of eVerolimus+irradiation grouP were significantly lower than those of irradiation grouP. The SER was 1. 36. The residual amount of γ_H2AX Protein in the eVerolimus + irradiation grouP was significantly higher than that of the irradiation grouP. Conclusion EVerolimus inhibiting mTOR signaling Pathway can increase the radiosensitiVity of A549 cells.