BACKGROUND:Chemokine receptor 4 (CXCR4) is the main homing regulating factor for mesenchymal stem cells in vivo. How to improve the self-expression of CXCR4 is crucial for regulating the homing ability of stem cells in vivo targeting organs. OBJECTIVE:To construct lentiviral vectors carrying CXCR4 or green fluorescent protein (GFP) gene, and to observe their effects on growth and secrete function of bone marrow mesenchymal stem cells. METHODS:Lentiviral vectors were used to over-express CXCR4 in rat bone marrow mesenchymal stem cells. GFP gene was transfected as a control. Transfection efficiency was analyzed using fluorescence microscopy, RT-PCR and western blot assay for detecting the expression of CXCR4 or GFP in cells at both the mRNA and protein levels. Effects of CXCR4 expression modified on the cellular proliferation of bone marrow mesenchymal stem cells were assayed with flow cytometry. Effect of CXCR4 regulation on secretion function of bone marrow mesenchymal stem cells was determined by protein analysis from cellsupernatant. RESULTS AND CONCLUSION:CXCR4 gene or GFP gene could be transfected into bone marrow mesenchymal stem cells safely and effectively with lentivirus transfection system. Analysis of CXCR4 expression at the mRNA and protein levels confirmed the transfection efficiency. Effects of CXCR4 over-expression on the cellular proliferation of bone marrow mesenchymal stem cells and effects of CXCR4 regulation on secretion of protective factors from bone marrow mesenchymal stem cells were improved. The proliferation ability of bone marrow mesenchymal stem cells with CXCR4 over-expression was improved, which was approved with flow cytometry (P<0.05). And it was helpful for bone marrow mesenchymal stem cells to secrete some protective factors. Many protein and soluble factors were at higher levels, and those were good at cells proliferation and immunoloregulation in bone marrow mesenchymal stem cells (P<0.05). The method of lentiviral transfection is a safe and effective way to modify bone marrow mesenchymal stem cells. We can improve the ability of bone marrow mesenchymal stem cells in the survival and emiocytosis ability through genetic engineering.

Objective IFN-? is a pleiotropic cytokine with potent immunomodulatory effects and antiviral activity. To study the mechanism of IFN-? clearing duck hepatitis B virus (DHBV) in ducks, it is essential to establish a method to quantify expression of DuIFN-? in immune response. In the present study,a semi-quantitative competitive RT-PCR was developed to quantify expression of duck IFN-?(DuIFN-?) mRNA by PBMCs. Methods Based on ?-actin consensus sequence, fishing the ?-actin gene as house keeping gene from duck PBMC by RT-PCR. A competitive internal control was constructed and the competitive RT-PCR system could be used to quantify the transcription of DuIFN-? mRNA. Results After duck PBMCs were stimulated in vitro with PHA, the peak of DuIFN-? expression was at 24-36h. Then RT-PCR method was applied to detect DuIFN-? mRNA transcription by PBMCs from DHBV infected ducks immunized with DuIFN-? plasmid plus DNA vaccine or DNA vaccine alone. Results showed that expression of DuIFN-? in ducks co-immunized with DuIFN-? plasmid were higher than other groups immunized without DuIFN-? plasmid as adjuvant. Conclusions The results indicated that DuIFN-? gene could be a useful adjuvant to develop vaccines. The semi-quantitation of DuIFN-? mRNA by competitive RT-PCR provides the basis for future study of the mechanism of IFN-? in duck hepatitis B virus persistent infection.

Objective To explore the methods for treating no-functioning renal graft.Methods The management of 240 cases of non-functioning renal graft between 1978 and December 2003 was retrospectively analyzed.Results Twenty-nine recipients with renal failure one month after renal transplantation were subjected to removal of the renal graft. One year after renal transplantation, the recipients with renal failure received resection of the renal graft, 4 were treated with renal artery embolism, and 114 did not receive special treatment.Conclusion The non-functioning renal graft required removal at early stage of renal transplantation. At later stage, no symptomatic cases of post-transplanted renal failure required no management after stopping immunosuppressant. The appropriate management is reserved for the symptomatic cases.

BACKGROUND: Anemia after kidney transplantation has a clinical incidence rate of 30%-40%, is the important risk factor for cardiovascular diseases and kidney failure after kidney transplantation and is also the independent prediction index of patient's death. OBJECTIVE: To analyze the factors related to anemia after kidney transplantation. RESULTS AND CONCLUSION: In the anemia group (n = 225, 27.2%, aged 26-65 years), the incidence rate of anemia in female and male patients was 23% and 37%, respectively (P < 0.05), 46 patients had hypertension and used angiotensin-converting enzyme inhibitor or angiotensin Ⅱ receptor antagonist and 16 patients had chronic erosive gastritis or upper gastrointestinal tract ulcer, with the human survival rate of 85.3% and kidney failure rate of 25.3%. In the non-anemia group (n = 601, 72.8%, 405 males, 196 females, aged 18-71 years), 35 patients had hypertension and used angiotensin-converting enzyme inhibitor or angiotensin Ⅱ receptor antagonist and 14 patients had chronic erosive gastritis or upper gastrointestinal tract ulcer, with the human survival rate of 92.1% and kidney failure rate of 12.6%. There was significant difference in above-mentioned indices between anemia and non-anemia groups (P < 0.05). These results suggest that gender, age, kidney function, digestive tract disease history, and drug application are closely related to anemia after kidney transplantation.

BACKGROUND:Autologous peripheral blood hemopoietic stem cell transplantation(HSCT)in combination with high-dose chemotherapy significantly improves complete remission and survival rate of multiple myeloma patients.However,the relapse rate is high.Bortezomib is 26S proteasomes inhibitor,and effective on the primary treatment of multiple myeloma.OBJECTIVE:To evaluate the curative effect of HSCT in combination with bortezomib and high dose-melphalan for multiple myeloma.METHODS:A retrospective analysis of 3 patients with a stage-ITT multiple myeloma admitted to Department of Hematology,Affiliated Hospital of Guiyang Medical College from October 2006 to May 2007,was conducted.Chemotherapy and granulocyte colony-stimulating factor were used to mobilize autologous peripheral blood hemopoietic stem cells.All patients were pretreated with 200 mg/m2 melphalan via intravenous drip 3 days before transplantation,followed by HSCT 48 hours after drug termination.RESULTS AND CONCLUSION:All patients obtained prompt and sustained hematopoietic reconstitution,and bone marrow depression restored 30 days following HSCT.Case 1 and 2 obtained complete remission,and case 3 obtained partial remission.Results show that HSCT in combination with bortezomib and high-dose melphalan is a safe and feasible treatment on multiple myeloma.The patients have good tolerance to pretreatment.

BACKGROUND: Rituximab single or in combination with CHOP regimen for treatment of CD20-positive non-Hodgkin lymphoma has achieved good curative effects. Autologous hematopoietic stem cell transplantation (AHSCT) has been shown to improve the curative effects and increase survival rate of patients with non-Hodgkin lymphoma. However, the curative effects of these two methods remain disputed. OBJECTIVE: To investigate the efficiency of rituximab in combination with AHSCT on CD 20-positive non-Hodgkin lymphoma. METHODS: Six patients with CD 20-positive non-Hodgkin lymphoma (stage IV) underwent AHSCT and rituximab administration. 375 mg/m~2 rituximab was intravenously administered 2-4 times prior to AHSCT, twice prior to and after peripheral blood stem cells mobilization and preprocessing, respectively, as well as once every 3 months after AHSCT. RESULTS AND CONCLUSION: The mean number of mononuclear cells and CD 34-positive cells was 5.13×10~(-8)/kg and 4.75×10~(-6)/kg, respectively. Following AHSCT, all 6 patients presented normal hematopoietic functions, neutrophils exceeded 0.5×10~(-9)/L at 9-15 days and blood platelet counts exceeded 20×10~(-9)/L at 12-19 days. Hemorrhagic cystitis, interstitial pneumonia, cytomegalovirus infection, or hepatic venous obstruction was not observed during the whole process of AHSCT in each patient. At 6-32 months, patients completely recovered. These results indicate that rituximab in combination with AHSCT is a good method for treatment of CD20-positive non-Hodgkin lymphoma and rituximab maintenance therapy could prevent disease recurrence.

Objective To explore the indication,surgical procedure and the postoperative treatment of breast conserving modified radical mastectomy for early stage breast cancer .Methods The clinical data of 21 patients with early stage breast cancer underwent breast conserving modified radical mastectomy were analyzed retrospectively. Results All patient recovered uneventfully. All the external configuration of the breast were fine. There were no recurrence and no complications in this series. Conclusions The breast conserving modified radical mastectomy is recommended for the early stage breast cancer. The external configuration of breast is fine postoperatively, and patients have higher survival quality.But follow up is necessary in order to find and treat the recurrence of breast cancer.

Objective To investigate the effects of CSK-conjugated PLGA nanoparticles on oral delivery of insulin in vitro and in vivo.Method CSK-INS-NPs were prepared by double-emulsion. Nanoparticle size、zeta potential and entrapment efficiency were measured. The efficiency of cellular uptake on Caco-2 cells in vitro was evaluated. The hypoglycemic effects were evaluated by monitoring the glucose levels in diabetic rats. Results The average sizes was(134. 4 ± 15)nm and their PDI values were less than 0.3.The insulin entrapment efifciency was around 71%. The cellular uptake of CSK-INS-NPs by Caco-2 cells was 2.8 times higher than INS-NPs. The CSK-INS-NPs transferred more insulin across the Caco-2 cell monolayer than INS-NPs and insulin solution did. In vivo experiments,the CSK-INS-NPs could reduce the blood glucose level of diabetic rats after oral administration in 10 h.Conclusion Compared with insulin solution,CSK-INS-NPs enhanced the insulin through Caco-2 cell monolayer by transcellular pathway and may be a potential delivery system for Oral Delivery of Insulin.

OBJECTIVE:To establish a New Vierordt method for assaying ingredients in compound fleroxacin suppositories and compound fleroxacin ear drops METHODS:New Vierordt method was applied to determination of fleroxacin and tinidazole in compound fleroxacin suppositories and fleroxacin and metronidazole in compound fleroxacin ear drops with no need of separation RESULTS:The fleroxacin and tinidazole in compound fleroxacin suppositories were detected at wavelength 286nm and 317nm respectively The average recoveries were 100 17% and 99 96% with RSDs=0 44% and 0 37% respectively The fleroxacin and metronidazole in the compound fleroxacin ear drops were detected at wavelength 286nm and 318nm respectively The average recoueries were 100 65% and 99 92% with RSDs=0 65% and 0 21% respectively CONCLUSION:The New Vierordt method can be used for quality control of the two compound preparations because of its convenience,accuracy and good repeatability and can eliminate mutual interference of two ingredients in preparations

OBJECTIVE:A HPLC method was established for determination of the level of sodium ferulate in plasma.ME_THODS:A C18 radial compression column was used with a mobile phase of CH3OH∶H2O∶H3PO4(250∶250∶0.2).The detecing wavelength was 254nm.RESULTS:The linear range was 1.2～24?g/ml(r=0.9 993).The average recovery was(98.8?0.6)%.The RSDs of within-day and between-day were less than 10%.CONCLUSION:This method provides an accurate and sensitive way in detecting plasma concentration of sodium ferulate.