Objective:To investigate the inhibitory effect of recombinant human endostatin(ES)combined with rAd/P53 on transplanted human breast cancer cell MCF-7 in nude mice.Methods:Animal breast cancer model was established by inoculating MCF-7 cells in nude mice.The animals were randomized into control group,ES group,rAd/P53 group,and ES+ rAd/P53 group.The tumor growth was observed in each group and immunohistochemical method was employed to examine the microvessel density(MVD) and the expression of vascular endothelial growth factor(VEGF).Results:Tumor growth was significantly inhibited in all the 3 treatment groups(P

Background and purpose:There is a tendency to treat tumors through multiple genes and multiple targeting but there has been no reports about the therapeutic effect of recombinant human endostatin(ES)combined with rAd/p53 on graft model of human breast cancer.Methods:MCF-7 human breast cancer cells were inoculated in nude mice.Then the nude mice with xenograft tumor were randomized into groups of control,ES,rAd/p53,and ES + rAd/p53,they were treated according to the plan.Diversity of the xenograft tumor volume and side effect was observed.Microvessel density(MVD)and expression of vascular endothelial growth factor(VEGF)were detected by immunohistochemistry.Results:After the administration of ES,rAd/p53,ES + rAd/p53,the growth of tumor was inhibited with the inhibitory rate of 1.75%,43.36% and 58.68%,respectively.Inhibition of tumor growth was significant in Group rAd/p53 and ES + rAd/p53(P0.05).Conclusions:recombinant human endostatin combined with rAd/p53 can greatly inhibit growth of xenograft tumor,and reduce the generation of capillary vessels.

AIM:To explore the feasibility and biological characterization of long-term regulated expansion of JAK2 transduced human CD34+ cord blood cells in vitro.METHODS: A retrovirus (RV) vector which contains JAK2 catalytic domain and two binding sites for a chemical inducer, dimerization (AP20187), was cloned (designated MGI-F2JAK2). CD34+cells were enriched from cord blood with a MiniMACS system. The purified CD34+cells were transfected with supernatant from the retrovirus packaging cell line that expressed JAK2. Following transduction, cells were expanded into four groups: AP20187 alone, FL alone, TPO, alone, AP20187+FL+TPO, respectively. The expanded cells were monitored by GFP expression, immunophenotyping, progenitor colony assay, karyotype analysis as well as tumorigenesis in nude mice. RESULTS: The purity of selected CD34+ cells was over 91% and gene transfer rate was 49.32%?6.21%. Only the group of AP20187 +FL+ TPO was obtained a significant sustained outgrowth of the transduced CD34+ cord blood cells. The percentage of GFP+ cells consistently produced a rise to the 90% peak level by the end of 8th week of culture. Flow cytometry analysis showed that the phenotype of the expanded cells was CD33+, CD61+ and Gly-A+ partial positive; CD38+ and HLA-DR+ strong positive, while CD2, CD7 and CD19 were almost negative. Colony assays performed in methycelluos, which can give rise to BFU-E, CFU-GM and CFU-Mix, the CFU-GM was predominantly in all colonies. The tumor was not observed in nude mice and the karyotype analysis was normal from expanded cells.CONCLUSION: The results demonstrate that AP20187-mediated activation of JAK2 signaling is capable of stimulating expansion JAK2 transduced CB CD34+ cells in combination with FL and TPO. This system may have applications for studies in signaling transduction, hematopoiesis, and for gene and cell therapy.

Objective To explore the immunophenotype of chronic lymphocytic leukemia (CLL). Methods The immunophenotypes of 31 patients with CLL were determined by immunocytometry. Results Among 31 cases with CLL, the positive expression rates of CD19,HLA-DR, CD5, CD23, CD20, CD22, CD38, FMC7 and CD10 were 100%, 96.8%, 90.3%,90.3%, 83.9%, 54.8%, 32.3%, 6.5% and 0.0%, respectively. Conclusions The imrnunophenotype analysis is very important for diagnosing CLL and it can early detect monoclonal B-cell lymphocytosis of CLL, which is the early phase of CLL, and provide early warning for patients.

Objective To evaluate the characteristics of immunophenotyping in adults with acute myeloid leukemia (AML) using muhi-eolor flow cytometry. Methods Immunophenotyping was performed by three color flow cytometry using CD45/SSC gating. Results In 126 patients with AML, the myeloid antigen of CD13, CD33 and CD117 was highly expressed. The positive rate was 86.4 %, 70.2 % and 90.4 %, respectively. The CD34 and HLA-DR were lowerly expressed as 63.5 % and 61.7 %, respectively. About 34.2 % of lymphoid-assoeiated antigen expression of all the AML patients. The lymphoid-associated antigens of CD7 and CD19 expression in patients with AML was 23.6 % and 2.3 %, respectively. Conclusion Multi-color flow cytometry is an important method for diagnosis and prognosis for AML.

Objective To explore the procedures of 3Q validation for blood coagulation analyzer under the good laboratory prac‐tice(GLP) system based on the Sysmex CA7000 automatic blood coagulation analyzer .Methods Four test indicators of PT ,APTT , TT and FIB were chosen to conduct the performance verification in the within‐run precision ,between‐day precision ,accuracy ,linear‐ity ,carry‐over contamination rate of the automatic blood coagulation analyzer .They were prothrombin time(PT) ,activated partial thromboplastin time(APTT) ,thrombin time(TT) ,and fibrinogen(FIB) .Results The within‐run precision CV values of PT , APTT ,TT and FIB were 1 .33% ,1 .57% ,1 .47% and 1 .90% ,respectively ;the inter‐day precision CV values of PT ,APTT ,TT and FIB were 1 .73% ,1 .52% ,1 .55% and 2 .14% respectively ;in terms of accuracy ,the normal quality control CV values were 7 .45% ,3 .88% ,-4 .98% and 4 .36% respectively ;the abnormal quality control CV values of PT and APTT were 8 .11% and 8 .77% respectively ;the r value of FIB linear correlation coefficient was 0 .999 3 ,the a value was 1 .02 ;the highest CV value of car‐ry‐over contamination rate was 2 .15% ;the detected results all conformed to the general industry standards and requirements of in‐strument manufacturer .Conclusion The 3Q validation under the GLP system proves that the Sysmex CA7000 automatic blood co‐agulation analyzer is good in performance and is appropriate for the laboratory work of clinical laboratory department .

Objective To study the effects and its probable mechanism of ultrasound-irradiation docetaxel-loaded microbubbles on apoptosis in human liver cancer cells MHCC97-H.Methods The MHCC97-H cells were randomly divided into 4 groups:control group,docetaxel-loaded microbubb[es group(DM),Ultrasound group (US),ultrasound irradiating docetaxel-loaded microbubbles group (DM + US).The livability,the morphological change of cells,cell apoptosis rate and the expression of proliferating cell nuclear antigen and MAPK pathway members of each group were respectively detected.Results After 24h,MTT test showed that the livability of MHCC97-H cells of DM group,US group and DM + US group were (76.0±3.0)％,(73.0± 5.0)％ and (52.0 ± 6.0)％ respectively,which was significantly declined comparing with control group (P ＜ 0.05).The morphological of tumor cells in control group remain normal size and grow eugenic,while the cells of DM group,US group and DM + US group decreased,their shapes were irregular,the intercellular space increased,and cells apoptosis occured in the medium.The cells in DM + US group were changed significantly.FCM showed the apoptosis rate of DM + US group was (26.81 ± 2.82) ％,which was declined clearly compared with (2.90 ± 0.99) ％ of the control group (P ＜0.05).Western-blot showed that the expression of proliferating cell nuclear antigen and p-ERK1/2 protein in DM + US group were down-regulated,and meanwhile p-p38 protein in human liver cancer cells MHCC97-H was up-regulated.Conclusions The method of ultrasound irradiating docetaxel-loaded microbubbles can significantly induce human liver cancer cells MHCC97-H apoptosis by regulating the expression of related protein.

Objective Preoperative autologous blood donation(PABD) can reduce the demand of allogeneic blood transfusion and its safety in obstetrical application has been proved.The article aimed to explore the effects of PABD on reducing allogeneic blood transfusion in pregnant women with placenta previa and the optimal PABD volume for implanted placenta.Methods Retrospective analysis were made on 156 cases with placenta previa hospitalized in our hospital from January 2015 to April 2016, including 78 cases with placenta implantation.According to the volume of PABD, the cases were classified into no PABD group, 300~400mL PABD group, and 600ml PABD group.Data of postpartum hemorrhage volume and allogeneic blood transfusion after delivery were collected to analyze the effectiveness of PABD in reducing the need for allogeneic blood transfusion during pregnancy.Results The hemorrhage volume during the delivery of all 156 patients with placenta previa was 230-5670mL (median 985ml), the rate of severe postpartum hemorrhage (PPH) was 49.4% (77/156), and the rate of allogeneic blood transfusion was 33.3% (52/156).In patients who had no PABD, the rate of allogeneic blood transfusion was 48.2% (40/83).However, this rate dropped down to 16.4% in PABD patients (12/73)(χ2=17.624,P<0.001).The rate of allogeneic blood transfusion in patients was different according to the situation of placenta planting, 43.3% in patients with no placenta plantingand 53.8% in patients with placenta planting.600ml autologous blood could meet all the needs for blood transfusion if there was no placenta implantation.300-400mL PABD could meet the needs of more than 80% patients.11.2%-13.3% of ABD patients might need allogeneic blood transfusion in addition to autologous blood.However, the amount of allogeneic RBC and FFP per capita reduced.Conclusion Patients with placenta previa is in high risk of PPH and PABD can improve their medical safety by reducing the rate and volume of the allogeneic blood transfusion.The strategy of 300-400mL PABD during pregnancy are recommended if there is no contraindication.

Objective To explore the effect of uremic serum on apoptosis in human umbilical vein endothelial cells(HUVECs) and the intervention by recombinant human erythropoietin(rhEPO) in this process.Methods From Dec.2008 to Apr.2009,10 uremic patients and 10 healthy volunteers were enrolled in this study.HUVECs were divided into 3 groups:control group(including 10% healthy serum medium),uremic group(including 10% uremic serum medium) and rhEPO treatment group(rhEPO at 5,10 or 15 U/ml was added to 10% uremic serum medium).After 24 hous's intervention,cell apoptosis was evaluated by TUNEL assay,and reactive oxygen species were detected by colorimetry.Results HUVECs apoptosis index and ROS level were higher in the presence of uremic serum than those of healthy serum(P

Background and purpose:Multidrug resistance(MDR)is the main obstacle of chemotherapy in the treatment of cancer,and it has been reported that the muted p53 gene is related to MDR.In this study,we explored whether adenovirus mediated p53 gene(Ad-p53)could reverse MDR of human breast cancer and its impacts on the expressions of P-gp,TOPOⅡ and GST-?.Methods:In this study,adriamycin-resistant human breast carcinoma cells(MCF-7/Adr)and its parental cells(MCF-7)were used to determine the effect of Ad-p53.Cck-8 assay was adopted to evaluate the cytotoxicity of adriamycin.Western blot were performed to observe the expression of P-glycoprotein(P-gp),TopoismeraseⅡ(TOPOⅡ)and GST-?.Results:After transfection with 50 MOI Ad-p53,the 50% inhibitory concentration(IC50)of adriamycin to MCF-7/Adr cells was decreased from(4.54?0.91)?g/ml to(0.26?0.11)?g/ml,and the chemosensitivity increased 18.1 times(P