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.

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.

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 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

Objective To investigate the effects of local application of NGF combined with Insulin on wound healing with Ⅱ degree deep scald and the expression of HIF-1α and VEGF in diabetic rats. Methods 60 male Wistar rats were used in the study by intraperitoneal injection of streptozotocin (STZ). Deep partial thickness scalding was engendered on the back of the rats after one month. Then these rats were random divided into group B (scalding of the rats with diabetes, n = 15), C (insulin control, n = 15), D ( NGF control, n = 15) and E ( NGF and insulin supplementation, n = 15). And group A ( nomal control,n = 15) was created by normal rats of partial thickness scalding. The wound area, wound healing rate, and grey of HIF-1α and VEGF immunohistochemistry staining were determined on the 3rd, 7th, 15th, and 21st day post scalding days (PSDs). Results Compared to those in groups A, B, C and D, the wound healing rate in group E increased significantly since the 7th PSD [(25.33 ± 2. 32 ) %, P ＜ 0.05]. The wound healing rate in group C and D increased significantly compared with group B [( 16. 68 ± 1.95 ) %, ( 18.29±1. 70)%, P ＜0.05]. Compared to group A, the group D increased significantly since the 15th PSD [(54. 84 ±3.60)%, P ＜0. 05]. The expression of HIF-1α and VEGF in group E began to increase slower than other groups after the 7th day ( P ＜0. 05 ). Conclusion Local application of NGF combined with Insulin could be beneficial to the regeneration of capillary vessel in the burn wounds of the rats with diabetes,as well as to accelerate wound healing by redress anoxia and decrease the expression of HIF-1α and VEGF.

Objective To investigate the effect of endoplasmic reticulum stress in renal damage caused by hyperlipidemia. Methods Forty male Wistar rats were randomly divided into two groups: normal control group (NC, n=20) and high-fat group (HF, n=20). Rats in NC group were fed with normal diet while those in HF group were fed with high-fat diet. Five rats in each group were randomly chosen in week 4, 8, 12 and 18. Serum lipid, urine protein in 24 hours and the pathological changes of renal tissues were observed; the apoptosis of renal cells was detected by TUNEL staining; the expression of GRP78 protein in the kidney was examined by immunohistochemistry. The expression of GRP78 mRNA and CHOP mRNA was examined by RT-PCR. Results Compared with those in NC group, serum lipid as well as the expression of GRP78 mRNA and protein in the kidney were increased in week 4, 8, 12 and 18 in HF group(P<0.05). In contrast, urine protein in 24 hours, the apoptosis index of renal cells and the expression of CHOP mRNA were increased in week 8, 12 and 18 (P<0.05). Conclusion CHOP pathway of endoplasmic reticulum stress is involved in renal damage caused by hyperlipidemia.

AIM: To investigate the role of endoplasmic reticulum stress in renal injury caused by hyperlipidemia and the influence effect of simvastatin. METHODS: Thirty male Wistar rats were randomly divided into three groups: rats in control group (n=10) were fed with normal diet; rats in high fat group (n=10) were fed with high fat diet; animals in simvastatin+high fat group (n=10) were fed with high fat diet and were received simvastatin 10 mg·kg~(-1)·d~(-1) by gastric irrigation. After 18 weeks, the quantitative urine protein in 24 h, the serum cholesterol and triglycerides levels were tested. The pathological changes of renal tissue were observed under optic microscope. The expressions of GRP78 and p-JNK in renal tissues were examined by immunohistochemistry. The apoptotic cells in the kidney were detected by TUNEL staining. The mRNA expressions of GRP78 and CHOP were examined by RT-PCR. RESULTS: The quantitative urine protein in 24 h, the serum lipid, the expressions of GRP78 and p-JNK proteins, the mRNA expressions of GRP78 and CHOP as well as the apoptotic cells in renal tissues were increased in high fat group (P<0.01).The quantitative urine protein in 24 h, the serum lipid, the expression of GRP78 and p-JNK proteins, the mRNA expressions of GRP78 and CHOP as well as the apoptotic cells in renal tissues were remarkably reduced in simvastatin+high fat group than those in high fat group (P<0.05). CONCLUSION: The endoplasmic reticulum stress is engaged in the renal injury caused by hyperlipidemia. The simvastatin play a role in renal protection by inhibiting the endoplasmic reticulum stress in the kidney.

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.