Objective To identify,construct and express scFv CD133,verify its biological function.Methods VL and VH were isolated from hybridoma of mAb CD133 by using antibody engineering technology.Its DNA sequencing and CDR were determined.scFv CD133 was then cloned into pET32a,transformed into Origami,induced by IPTG,purified by Ni2+-NTA His resin.Its affinity and specificity were tested by NH4SCN elution and ELISA.Results The size of VL and VH of scFv CD133 was 339 bp and 342 bp,which coded 113 and 114 amino acid separately.Its VL belonged to mouse Igκ chain and VH belonged to mouse IgG heavy chain subtype I.The molecular weight of scFv CD133 was about 27 × 103 which was testified by SDSPAGE and Western blot.Its affinity and specificity were also verified.Conclusion scFv CD133 has been successfully constructed and expressed in Origami,which could supply basis for target therapy of CD+133 cancer stem cell.

To observe the inhibitory effects of puerarin on angiopoiesis of endometriotic tissue, and to explore the regulatory effects of puerarin on tumor-related gene expression of endometriosis.

Objective We evaluated the role of a quality improvement initiative in improving clinical outcomes in peritoneal di-alysis ( PD) . Methods In a retrospective analysis of 6 years of data from a hospital registry, the period between 1 July 2005 and 30 June 2008 ( control group) provided baseline data from before implementation of systemic outcomes monitoring, and the period between 1 July 2008 and 30 June 2011 [continuous quality improvement (CQI) group] represented the time when a CQI program was in place.Per-itonitis incidence, patient and technique survival, cardiovascular status, causes of death, and drop-out were compared between the groups. Results In the 370 patients of the CQI group and the 249 patients of the control group, the predominant underlying kidney diseases were chronic glomerulonephritis and diabetic nephropathy.After implementation of the CQI initiative, the peritonitis rate de-clined to 1 episode in 77.3 patient-months from 1 episode in 22.6 patient-months.In the CQI group, the complicance of blood pressure was more significantly improved than the control group ( 67.8% vs 47.4%,P<0.05).During the 3 years of follow-up,cardiothoracic ratio and IVST were significantly increased in the control group [0.55 ± 0.08 vs 0.51 ±0.05,P<0.05,11.07 ±1.66 mm vs 10.25 ±1.38 mm, P<0.05〗.The difference of LVID between the two groups was signifi-cant at the 2nd and 3rd year of follow-up(P<0.05).Patient survival at 1, 2, and 3 years was significantly higher in the CQI group (97.3%, 96.3%, and 96.3%respectively) than in the control group (92.6%, 82.4%, and 67.3%respectively, P<0.001).Imple-mentation of the CQI initiative also appeared to significantly improve technique survival rates:95.6%, 92.6%, and 92.6%in the CQI group compared with 89.6%, 79.2%, and 76.8%in the control group (P<0.001) after 1, 2, and 3 years respectively. Conclusion Integration of a CQI process into a PD program can significantly improve the quality of therapy and its outcomes.

Aim To study the effects of polyamine analogue CPENSpm on the human lung cancer line A549 in cell proliferation and apoptosis.Methods MTS was used to assay the cell proliferation,chemical analysis methods were used to determine the activities of enzymes in the polyamine metabolism,HPLC was performed to assay the intracellular concentration of polyamines,Sub-G1 and DNA fragmentation assays were used to determine the cell apoptosis.Results Treating A549 lung cancer cells by CPENSpm resulted in:①cell-growth inhibition and cell apoptosis;②inhibition of ODC(key enzyme in polyamine synthetic pathway)and activation of SSAT and SMO(key enzymes in polyamine catabolism);③great decrease of intracellular polyamine concentrations.MDL72527,the SMO inhibitor,can antagonize the effect of CPENSpm on inhibiting the proliferation of A549 cells.Conclusion CPENSpm inhibits proliferation and induces apoptosis of human A549 lung cancer cell line by interfering the polyamine metabolism,depleting intracellular polyamine contents that are need by quick-growth of cancer cells and inducing production of H2O2.

Objective To evaluate the value of MR and CT examinations in the diagnosis of nasopharyngeal carcinoma (NPC) and compare 2008 staging system with 1992 staging system and 2002 UICC staging system for NPC. Methods MR and CT images of seventy-six cases with NPC were studied. According to 2008 staging system and taking MR as a standard, differences between these two examinations were evaluated under the new NPC staging system, and three staging system were compared by MR findings. Results MR was inconsistent with CT in eveluating invasion of medial pterygoid muscle(22,24 cases), lateral pterygoid muscle( 15, 11 cases), skull base(35, 32 cases) and intracranial fossa( 11,6 cases), but no statistical diffence existed ( P ＞ 0. 05 ). There were statistical difference ( P ＜ 0. 05 )between MR and CT in determining invasion of parapharyngeal space( 50, 61 cases), retropharyngeal lymph node metastasis(48, 23 cases), stage T1 (18, 11 cases), T2 (15, 22 cases), N0 (18, 24 cases) and N1(33, 27 cases) with differences of 11 cases, 25 cases, 7 cases, 7 cases, 6 cases and 6cases respectively.For invasion of parapharyngeal space, CT showed 11 cases more than MR while 5 cases were comfirmed as compression by local tumor and 6 cases were proved as retropharyngeal lymph node metastasis according to MR. For retropharyngeal lymph node metastasis, MR presented 25 cases more than CT. These two reasons above mainly caused the differences of T-staging and N-staging. For 2008 staging system, when compared with 1992 staging system, there were 9 cases upstaging and 1 case downstaging in T classification, 16 cases upstaging in N classification, and 15 cases upstaging and 1 case downstaging in clinical classification; and when compared with 2002 UICC staging system, there were 7 cases, 10 cases and 12 cases upstaging in T,N, and clinical staging respectively. Conclusions Compared with MR examination which was regarded as standard by 2008 staging system of NPC, there were some differences in demonstrating invasion of parapharyngeal space and retropharyngeal lymph node metastasis by CT. Compared to 1992 staging system and 2002 UICC staging system, 2008 staging system mainly made T and N classification of tumor upstage,resulting in upstaging in clinical classification.

Objective To evaluate the effect of preloading the epidural space with normal saline (NS) on the incidence of injury to blood vessel by epidural catheter placement for cesarean section. Methods One hundred and fifty parturients with a single baby at full term in vertex presentation scheduled for cesarean section under continuous epidural anesthesia were randomly divided into 3 groups ( n = 50 each): Ⅰ group control; Ⅱ group NS needle attached to a 5 ml syringe. Loss of resistance was used to identify the epidural space. In group Ⅰ no fluid was injected into the epidural space before insertion of catheter; while in group Ⅱ and Ⅲ NS 5 ml with or without whom blood or blood tinted fluid was withdrawn from epidural catheter was recorded. Results The number of patients in whom blood or blood tinted fluid was withdrawn from epidural catheter was significantly lower in group Ⅱand Ⅲ than in group Ⅰ but was not significantly different between group Ⅱ and Ⅲ. Conclusion Preloading the epidural space with 5 ml NS can reduce the incidence of injury to blood vessel induced by insertion of epidural

Objective To investigate the value of DWI before treatment on predicting sensitivity of concurrent chemoradiation in nasopharyngeal carcinoma.Methods Seventy patients with nasopharyngeal carcinoma proved by nasopharyngoscope and biopsy pathology conducted DWI before concurrent chemoradiation and reexamined on receiving dose of 50 Gy.The mean, maximum and minimum ADC value of tumor were measured on DWI and maximum area of tumor before and during treatment ( on dose of 50 Gy) was delineated to calculate the tumor regression rate ( RS0-50 ).The patients were classified into three groups according to the RS0-50 as sensitive, moderate, and resistant therapeutic effect.Patients were classified into different groups according to the pathologic type and clinical stage respectively .Spearman correlation analysis was used between RS 0-50 and ADC values of all tumors , different pathologic types and clinical stages , respectively.ROC was used to evaluate the cutoff and value of ADC which had highest correlation to RS0-50 on predicting therapeutic effect.Results DWI of 3 patients were excluded due to obvious swallow artifact which influenced the measurement , and finally 67 patients were included in this study, with pathological type of nonkeratinized differentiated undifferentiated carcinoma in 49 cases, nonkeratinized undifferentiated carcinoma in 18 cases, clinical T1 stage in 7 cases, T2 in 14 cases, T3 in 17 cases and T4 in 29 cases.During treatment , there were 13 cases with sensitive therapeutic effect , 42 cases with moderate therapeutic effect and 12 cases with resistant therapeutic effect.RS0-50 [ ( 65.6 ± 3.1) %] showed mildly and moderately negative correlation to mean ADC [(1.06 ±0.19) ×10 -3 mm2/s] and maximum ADC [(1.29 ±0.33) ×10 -3 mm2/s] respectively ( r =-0.276, P =0.024 and r =-0.434, P=0.001, respectively).ROC showed when setting threshold at maximum ADC value of lower than 1.06 ×10 -3 mm2/s for predicting sensitive therapeutic effect , the specificity , sensitivity , and accuracy was 69.2%(9/13), 88.9%(48/54) and 85.1% (57/67), respectively, and when setting threshold at maximum ADC value of higher than and equal to 1.30 ×10 -3 mm2/s for predicting resistant therapeutic effect, the specificity, sensitivity, and accuracy was 75.0% (9/12), 65.5% (36/55) and 67.2%(45/67), respectively.Conclusion Pretreatment maximum ADC value were able to predict the tumor regression rate and sensitivity of concurrent chemoradiation in nasopharyngeal carcinoma .

Objective To investigate the value of perfusion CT in the evaluation of neoadjuvant chemotherapy in locally advanced squamous cell carcinoma.Methods Sixty-seven patients with Ib2-IIb squamous cell carcinoma of the cervix underwent CT perfusion imaging before neoadjuvant chemotherapy to measure blood flow (BF),blood volume (BV),peak time (MTT),permeability sur-face (PMB),perfusion time to peak (TTP)and MIP(HU);and underwent routine enhancement CT after the two course of neoadju-vant chemotherapy treatment to measure tumor size,and evaluate the therapeutic effect.Results Among 67 patients,48 patents were effective (2 completely remission and 46 partial remission),and 1 9 patients were ineffective (1 5 stable and 4 progressed),with the overall response rate of 71.6%.The differences of PMB and BV value between effective and ineffective group were statistically significant before neoadjuvant chemotherapy (P <0.001).The BF,MTT,TTP value were no significant difference between effec-tive and ineffective group before neoadjuvant chemotherapy (P >0.05).The BV and PMB value were relatively higher in the patients with effective chemotherapy.Conclusion CT perfusion imaging could measure the BV and PMB value of the tumor before neoadju-vant chemotherapy to provide the basis for treatment selection.

OBJECTIVE To investigate the effect of furanodiene(FDE),a diterpene derived from the medicinal plant Zedoary,on apoptosis of human gastric cancer MGC-803 cells induced in vitro. METHODS MGC-803 cells were treated with FDE 46.29~740.74μmol·L-1 for 24,48 and 72 h,and the cell viability was detected with MTT assay. Cell morphology was observed by light microscopy and Hoechst33342 staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. Rh123 staining and fluorescence probe DCFH-DA were employed to detect the changes in mitochondrial membrane potential (MMP) and reactive oxygen species(ROS). RESULTS MTT Results showed that FDE 46.29-740.74μmol · L-1 exhibited significantly higher cytotoxicity to gastric cancer MGC-803 cells. IC50 for MGC-803 of 24,48 and 72 h treatment was 347.91,257.41 and 101.01μmol·L-1,respectively. Treatment with FDE 92.58-370.32μmol·L-1 for 24 h also caused significant morphological changes in MGC-803 cells. AnnexinⅤ-FITC/PI double staining showed that the apoptotic rate increased after FDE 92.58-370.32μmol·L-1 treatment for 24 h(P<0.05). FDE enabled MGC-803 cell cycle arrest in S phase. DCFH-DA staining showed that FDE resulted in an increase in intracellular ROS levels(P<0.05) when PDE concentration was 370.37μmol·L-1(P<0.05). MMP decreased after FDE treatment when PDE concen?tration was 370.37μmol·L-1(P<0.05). CONCLUSION FDE Possesses potent tumor selected toxicity and can induce apoptosis of MGC-803 cells through cell cycle arresting,which is related to inhibition of DNA biosynthesis.

Objective To investigate the effect of DNA-dependent protein kinase catalytic subunit (DNA-PKcs) on autophagy induction by ionizing radiation ( IR ) .Methods The cell model of knocking-down DNA-PKcs expression was constructed by transfecting HeLa cells with a pSicoR-based lentivirus vector expressing DNA-PKcs specific shRNA .Cellular growth activity and radiosensitivity were detected by cell countingkit ( CCK)-8 assay.The expression of autophagy related proteins was detected by Western blotting hybridization .Autophagy was also detected by monitoring the autophagic marker green fluorescere protein ( GFP )-light chain 3 ( LC3 ) puncta per cell under an immunofluorescent microscope .Results A cellular model of knocking-down DNA-PKcs expression was successfully generated by transfecting the specific shRNA against DNA-PKcs.Depression of DNA-PKcs significantly decreased the growth activity of HeLa cells and increased the cellular sensitivity to ionizing radiation .Both the expression changes of P 62 and LC3 proteins and immunofluorescent GFP-LC3 puncta observation indicated that knocking-down DNA-PKcs prompted the induction of autophagy by ionizing radiation . Moreover, inactivation of DNA-PKcs led to a decreased phosphorylation of mammalian target of sirolimus ( Rapamycin, RAPA) ( mTOR) at S2481 site.Conclusion Depression of DNA-PKcs expression prompts the induction of autophagy by IR and cellular radiosensitivity .mTOR signaling may be involved in the regulation of autophagy processing by DNA-PKcs.