Objective To evaluate the value of transcatheter arterial embolization(TAE) in the treatment of intractable epistaxis.Methods TAE using gelform or polyvinyl alcohol(PVA) particles of forty-one patients with intractable epistaxis were undertaken by the femoral artery approach,through selective catheterization of involved maxillary artery or the bleeding arteries for the stopage of bleeding..Results Of the forty-one patient,39 cases were cured by once TAE and the other 2 with recurrent bleeding on the next day after the TAE,to whom a second interventional treatment fullfilled the requirement.Conclusions Transcatheter arterial embolization is a simple,safe and effective treatment for the intractable epistaxis.

Objective To explore the endovascular therapeutic strategy for multiple occlusive lesions of aorta and iliac-femoral arteries, and to discuss the technical skill as well as the clinical significance. Methods A total of 8 patients with multiple occlusive lesions of aorta and iliac-femoral arteries were enrolled in this study. Preoperative CT angiography and MR angiography were performed in all the 8 patients. The lesions included complete occlusion of abdominal aorta below renal artery level (n = 2), distal abdominal aorta occlusion (n = 4), distal abdominal aorta stenosis (n = 1), distal abdominal aorta membranous occlusion (n = 1), and diseased iliac artery (n = 12), external iliac artery (n = 8), femoral artery (n = 1) and popliteal artery (n = 2). Endovascular interventional management, including opening channel, thrombolysis, balloon dilation, stent implantation, etc. was carried out via different routes. The results were analyzed. Results After endovascular interventional management the abdominal aorta was completely reopened in all the 8 patients. Of 12 diseased iliac arteries, 9 were successfully reopened by interventional treatment and the remaining 3 were not treated. All the diseased external iliac arteries were opened up. The involved femoral artery and popliteal arteries were not treated. The patients were followed up for 1 -12 months. During the follow-up period, ischemic symptoms of the lower limb disappeared in 5 patients and were obviously improved in 2 patients. Recurrence of thrombotic occlusion was observed in one case, which returned to normal after transcatheter thrombolysis therapy. Conclusion For the treatment of multiple occlusive lesions of aorta and iliac-femoral arteries, endovascular interventional management is safe, simple and effective with fewer complications. The ischemic symptoms of the lower limb can be significantly improved.

Objective To explore the clinical value of the interventional and comprehensive treatment of iliac-femoral venous thrombus(I-FVT).Methods 32 patients with I-FVT were underwent interventional therapy.First the filter was implanted into the inferior vena caval via opposite side of femoral vein,then the catheteres were implanted into the pathologic regions within the vena to process the emboluses,and balloon-directed extend when necessary.Results The procedure of treatment was successful in all patients.The iliac-femoral veins were patent,swollen and pain symptom of lower limb disappeared gradually after operation.The followed-up study the longest period for 30 months showed no severe complications and recurrence.Conclusion The interventional and comprehensive therapy is of ideal effect on treating I-FVT,occuring interventional and comprehensive treatment has ideally effect to cure the patients with I-FVT.

Objective To construct the T-cells and B-cells combined epitope peptide gene based on the LipL32,OmpL1 and LipL21 protein from Leptospira interrogans and E.coli expression system,and better understanding of the immunological activity of the recombinant protein. MethodsThe immunodomaint T- and B-cells combined epitopes of LipL32,OmpL1 and LipL21 were identified and used to synthetic a new gene and then construct its prokaryotic expression system.The expression of recombinant protein was determined by SDS-PAGE; MAT was used to determine the titer of the antiserum to L. interrogans standard strains of China ; Western blot and ELISA were used to identify the immunity activity of the recombinant protein.Results The synthetic gene was effectively expressed in E.coli BL21 ( DE3 ) strain and mainly presented in dissoluble protein.Western blot result showed that the expression protein react well with the antibodies from immunized rabbit by Leptospira or recombinant protein.ELISA and MAT results showed that the multiepitope protein could cross-react with different serogroup or serovar of Leptospira.Conclusion In this study,we successfully constructed the recombinant T- and B-cells combine epitope gene of leptospires and expressed it in E.coli.The recombinant protein had a good immune activity,and could cross-reacted with antibodies from different serogroups Leptospira infected patients.

Objective To investigate the effects of small interference RNA (siRNA) targeting HOXA9 on the proliferation and apoptosis of human acute monocytic leukemia U937 cell line.Methods Effective and specific siRNA oligo targeting HOXA9 was designed and compounded.It was transfected transiently into U937 cells by cationic liposome.The cells was divided into three groups:experimental group(siRNA targeting HOXA9 was transfected by liposome),negative control group (negative siRNA was transfected by liposome) and cell control group (add equal cells and medium).The expression of HOXA9 mRNA and protein were detected by reverse transcription PCR and Western blot.The cell proliferation was assessed by MTT.The apoptosis of each group were measured by Annexin V-FITC.Results Aftcr transfected by siRNA targeting HOXA9,the relative mRNA expression levels of HOXA9 in the experimental group,negative control group and cell control group were (22.980±0.548) ％,(82.371±1.517) ％ and (84.637±2.252) ％,respectively (P ＜ 0.05),and the relative protein expression levels were (50.377±2.773).％,(105.500±3.900) ％ and (111.392±3.905) ％,respectively (P ＜ 0.05).The inhibitory rates of cell proliferation and the apoptosis rates of the experimental group were significantly increased.The inhibitory rates of cell proliferation of 24 h,48 h and 72 h were (41.909±4.333) ％,(54.470±3.756) ％ and (65.835±1.024) ％,respectively,and the apoatosis rate was (26.800±2.081) ％.Compared with 2 controls,the experimental group differences had statistically significance (P ＜ 0.05).Conclusion siRNA targeting HOXA9 can effectively silence HOXA9 gene expression in U937 cell,suppress cell proliferation and induce cell apoptosis obviously,which providing experimental basis for clinical lenkemia therapy by targeting HOXA9 gene.

BACKGROUND: It is of great importance in improving the clinical effect of human islet allograft to study and design models of such large animals as pigs or primates preclinically.OBJECTIVE: To evaluate the effect of different doses of streptozotocin (STZ) on inducing diabetes type Ⅰ models of nonhuman primates.DESIGN, TIME AND SETTING: A contrast observational animal experiment was performed in the Cell Transplantation and Gene Therapy Center, the Third Xiangya Hospital of Central South University from October 2007 to December 2008. MATERIALS: A total of 21 adult male rhesus monkeys were divided into a 125 mg/kg STZ group (n =5), a 75 mg/kg STZ group (n=5) and a 50 mg/kg STZ group (n=11).METHODS: STZ weighed with regard to body mass of animals was prepared into 25 g/L STZ solution with buffer that was prepared in advance. After being filtered and degermed, the new-prepared STZ of 125 mg/kg, 75 mg/kg and 50 mg/kg were administered by intravenous injection into the experimental monkeys respectively, which took 1-5 minutes.MAIN OUTCOME MEASURES: Liver and renal function, glucose metabolism and histomorphological changes of animals during 1-16 weeks following administration.RESULTS: In 125 mg/kg STZ group, two rhesus monkeys died, in 8 hours following STZ administration, of serious hypoglycemia caused by severely damaged pancreas β cells; All rhesus monkeys in this group had got significantly increased liver transaminase, serum creatinine and urea nitrogen at week 1 following STZ administration, which reached a peak during 2-4 weeks; One rhesus monkey in this group showed severe shortage of endogenous trypsin and hyperglycemia irreversible by exogenous insulin following STZ administration, and finally died at day 13 following STZ administration due to the glucose metabolic disorder, ketoacidosis, liver and renal failure; The other two survivors in this group kept high level of liver transaminase,urea nitrogen and serum creatinine throughout the observation period. In 75 mg/kg STZ group, rhesus monkeys presented significantly increased liver transaminase, serum creatinine and urea nitrogen at week 1-2 following STZ administration; After 4 weeks following administration, their liver and renal function presented with abnormality of different degrees; One rhesus monkey in this group had got injured renal function, decreased power of resistance, eyelid edema, general dropsy and irreversible infected rump after injection of STZ, and finally died at the end of week 5 following administration; Another rhesus in this group presented with irreversible continuous hyperglycemia, inappetence and significantly decreased weight, and finally died ofsystemic failure at week 9 following administration. In the 50 mg/kg STZ group, renal function of monkeys were slightly affected, with a transient mild rise which return to the normal level by the end of week 4 following administration; Only 3 animals in this group appeared eyelid edema during 1-4 weeks following administration which disappeared afterwards.CONCLUSION: STZ of 50 mg/kg is possibly the optimal dose for inducing diabetes models in most rhesus monkeys.

Objective To investigate the effects of Livin antisense oligonucleotide (ASODN) on the proliferation and apoptosis of human leukemia (K562) cells. Methods Specific phosphorothioate ASODN and missense oligonucleotide (MSODN) target Livin mRNA were synthesized and transfected into K562 cells following cationic liposome. The proliferation inhibition of K562 cells was assessed by MTT. The apoptosis rate of each group was detected by Annexin V-FITC. The expression of Livin mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Results ASODN at a final concentration of 600 nmol/Lcould inhibit the K562 cells proliferation (IR) was (52.99t2.67) % and the expressions of Livin mRNA (ODR)was (59.75±3.24) %, the apoptosis rate was apparently increased [(36.89±1.08) %] (P ＜0.01); but the difference between Lip-MSODN group, Lip control group and cell control group was not statistically significant (P ＞0.05).Conclusion Livin ASODN may decrease Livin gene expression, suppress K562 cells proliferation effectively, and induce significant apoptosis of K562 cells.

Objective To evaluate the effect of cytokine-induced killer cells (CIKs) as an adoptive immunotherapy option for treatment of leukemia relapse after allo-hematopoietic stem cell transplantation (allo-HSCT).Methods Two cases of infusion of donor CIKs in patients with leukemia relapse after allo-HSCT were retrospectively analyzed.Patient one relapsed 986 days (+986d) after HLA-matched unrelated donor allo-HSCT.Applications of chemotherapy only resulted in short term remission,but allo-CIKs were successfully expanded from the patient's peripheral blood mononuclear cells of donor origin.Totally five cycles of CIKs infusion were infused as an alternative of adoptive immunotherapy.Patient two had recurrent in the + 158d after HLA-matched sibling alloHSCT.At + 204d and + 294d,two cycles of CIKs which were expanded from donor peripheral blood mononuclear cells were infused.Results One cycle of CIKs was given to patient one after the application of chemotherapy to reduce the tumor burden,and the patient successively achieved complete remission.Again after additional four cycles of CIKs infusion,consistent remission was maintained during the following seven months.Patient two who had relapsed disease posttransplantation,achieved cytological complete remission after withdrawal of immunosuppressants and undergoing chemotherapy combined with G-CSF mobilized stem cell infusion.However,at + 187d,the patient suffered from side-effect of acute graft versus host disease and extramedullary infiltration.The symptoms were alleviated markedly after one cycle of CIKs infusion at + 204d.Moreover,the pain disappeared after an additional infusion at + 294d.And up to the present,the bone marrow aspiration showed complete remission while the extramedullary disease vanished.Conclusion The use of CIKs in the treatment of leukemia relapse after allogeneic bone marrow transplantation can be feasible and well tolerated.

ObjectiveTo establish a novel rapid detection method based on PCR-single-strand-conformational polymorphism (PCR-SSCP) to determine mutation of streptomycin-resistance associated rpsL and rrs genes in isolates of Mycobacterium tuberculosis (MTB).MethodsStreptomycin-resistance of 112 MTB isolates was detected using the routine drug susceptibility test,and a special PCR-SSCP assay was established.The mutations of rpsL and rrs genes in streptomycin-resistant MTB isolates were detected by PCR-SSCP and PCR direct sequencing (PCR-DS) ; the results from two techniques were compared.Results All isolates had both rpsL and rrs genes.Fifty-two isolates (46.4％) were streptomycin susceptible,in which only 1 isolate showed abnormal PCR-SSCP fragments from rrs gene,and the specificity of PCR-SSCP was 98.1％ (51/52).Sixty isolates (53.6％) were streptomycin-resistant,in which 46 (76.6％) and 11 ( 18.3％ ) isolates presented the abnormal PCR-SSCP fragments of rpsL and rrs gene,respectively.One streptomycin-resistant isolate showed abnormal PCR-SSCP fragments from both rpsL and rrs genes.The sensitivity of PCR-SSCP was 93.3％ (56/60).ConclusionThe PCR-SSCP that established in this study is a specific and sensitive method for rapid detection of the streptomycin-resistance associated mutations in rpsL and rrs genes of MTB.

The animal experiment studies have demonstrated that hyperthermia is a strong teratogen to many kinds of animal with high incidence of neural tube defects(NTD).Epidermiological investigation showed that hyperthermia was closely related to the acencephaly and excenphaly in human being,but little was known about the distinct mechanism of NTD induced by hyperthermia.In order to study the developmental mechanism of NTD induced by hyperthermia,we observed the effects of hyperthermia on the neuroepithelial cells in primary culture.The neural tubes of the hamster embryos on 10 d after fertilization were obtained.the neuroepithelia were dissociated and then seeded at the density of 1×106 cells per well.The cells were divided into experimental and control groups randomly.The experimental groups were exposed to 42 C for 20 min,whereas groups exposed to 37 Cserved as control.After treatment of hyperthermia,the cells were incubated continuously at 37 C in a 95%air/5%CO2 humidified incubator,the culture was terminated after different intervals.Phase-contrast microscopy.scanning electron microscopy.transmission electron microscopy,MTT assay,agarose gel electrophoresis analysis,TUNEL detection,immunocytochemistryand image analysis were made.The results of the experimental groups indicated that the floating cells and apoptotic cells increased in number,the neurites of the cells were shortened even disappeared,the survival cells decreased in number,the ultrastructure of the cells demonstrated distinct abnormal changes,the function of mitochondria was impaired,the expressions of both bcl-2 and bax were abnormal.The above results suggest that hyperthermia may induce apoptosis of neuroepithelial cells,duringwhich bcl-2 and bax genes may play important regulatory roles.