Objective: To evaluate the effect of the active specific immunotherapy with leukemia vaccine in the malignant hematopoietic diseases. Methods: We established the animal models by inoculating C57BL/6 rats with FBL-3 erythroleukemia cells and prepared three types of tumor vaccine, which were administered on the rats respectively. The MTT colorimetric assay was adopted 2 and 4 weeks later to test the cytotoxicity of macrophage (MΦ) and that of cytotoxic T lymphocyte(CTL) derived from the rats injected with tumor vaccines, and compared the results with the control group. Results: With the growth of erythroleulemia cells in the rats, the cellular immunity was seriously depressed, and the inhibition of specific cellular immunity was later than that of non-specific cellular immunity. The tumor vaccine made from inactivated tumor cells, IFA and cytokines (rGM-CSF, rIL-2 and rIL-6), promote the cellular immunity of tumor burden rats, especially the specific cellular immunity more efficiently than that of tumor vaccine made from inactivated tumor cells and IFA, but the third vaccine made from inactivated tumor cells alone has no effect. Conclusion: The tumor vaccine made from inactivated tumor cells with addition of IFA and cytokines (rGM-CSF, rIL-2 and rIL-6) provides a promising future in the active specific immunotherapy against hematopoietic tumor.

Objective To study the disinfection efficacy of c ompound disinfectant of peracetic acid. Methods Suspended liqu id quantitative sterilization test and metal corrosion test were carried out wit h different concentrations of disinfectant compound. Results T he killing rate of Bacillus subtilis var.niger spores e xposed to peracetic acid 500 mg*L-1 or available chlorine 300 mg*L-1 for 15 minutes was 99.94% and 97.91% respectively, while that of the spores e xpos ed to the compound disinfectant containing both of them reached 100%. The influ ence of organic substances on the bactericidal efficacy of this compound disinfe ctant was less than that on the bactericidal efficacy of the single ingredient. The corrosive effect of the compound disinfectant on the metals was milder than that of peracetic acid, but heavier than that of dichlorodimethylhydantoin. Conclusions Disinfectant efficacy of compound disinfectant increase s clearly, while metal corrosiveness decreases.

Objective:To evaluate the cytotoxicity of macrophage of C57BL/6 mice treated with the prepared leukemia vaccine.Methods:Established the leukemia burden mice and prepared three types of leukemia vaccine,which were administered on the mice respectively.The MTT colormetric method was adopted 2 and 4 weeks later to measure the cytotxicity of M? derived from the mice accept immunotherapy or prevention,and compared with control group.Results:(1)With the growth of leukemia cells in the mice,the cytotoxicity of M? was seriously depressed;(2)The administration of tumor vaccine prepared from inactivated tumor cells,IFA and cytokines,such as rGM CSF,rIL 2 and rIL 6,promote the cellular immunity of tumor burden mice,more effciently than tumor vaccine made from either inactivated tumor cells and IFA or inactive tumor cells themselves along.Conclusion:The tumor vaccine simply prepared with inactive tumor cells,IFA and cytokines can active the non specific cellular immunity that represented as M?,for example non specific cytotoxicity,antigen present,immune surveillance and so on,these can build a base for active the T lymphocyte the next.The tumor vaccine provides a promising future in the therapy against hematopietic tumor.

The functional experimental teaching reform plays a very important role in raising the teaching resources,consummating the experimental teaching management system,enhancing the student to practice the ability and so on and has obtained the good effect.But it also faces some problems waiting urgently to be solved and further perfected,to which this article has made the analysis and the discussion from the management science angle.The goal lies in deepening the educational reform in functional experiment unceasingly to provide the model.

Objective: To observe the promoting blood circulation by removing blood stasis of Lubai Capsule(LBC)(Rhizoma Phragmitis, Radix Paeoniae Alba, Radix Saposhnikoviae, Flos Schizonepetae, etc.). Methods: Acute blood stasis rat models were established with swimming in iced water and sc adrenalin in order to observe the effect of LBC on blood rheology. Mesenteric microcirculatory disturbance rat models were also established with adrenalin in order to observe the effect of LBC. Clotting time was measured in vitro with prothrombin time(PT) and kaolin partial thromboplastin time(KPTT) kit in order to observe its effects. Results: LBC could decrease the whole blood and plasma viscosity, fibrinogen, erythrocyte sedimentation and aggregation ratio of blood platelets of rats, ease the sticky condition of blood stasis rat models and prevent from forming thrombus. It could also inhibit the constraction and slowing of blood flow of thin artery, the reducing of open capillaries and change of fluid condition caused by adrenalin and improve these phenomena. PT and KPTT could be increased obviously. Conclusion: LBC can significantly promote blood circulation by removing blood stasis, because of improving blood rheology and mesenteric microcirculatory disturbance and inhibit endogenous and exogenous coagulation system.

Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.
Bacteriophages/genetics ; DNA, Complementary/*biosynthesis ; *DNA, Neoplasm/biosynthesis ; DNA, Recombinant/biosynthesis ; *Gene Library ; Genetic Vectors ; Leukemia, Promyelocytic, Acute/*genetics ; Leukemia, Promyelocytic, Acute/metabolism ; Leukemia, Promyelocytic, Acute/pathology ; RNA-Directed DNA Polymerase/metabolism ; Transcription, Genetic/genetics

ObjectiveTo perform pharmacology problem-based learning (PBL) and evaluate its effects.MethodsPBL was performed for the clinical medicine class of grade 2007 and the satisfactory degree of students to teaching effects was observed with questionnaire. Results The students thought that PBL teaching had substantial contents and proper schedule and increased learning interest. Students' participating degree, mutual communication and controlling discussion procedure were fine,which reached the expected learning objective. ConclusionsThe effects of PBL teaching were excellent and most of our students could accept it.

Objective To investigate the expression level and clinical significance of WT1 gene in acute luekemia (AL) patients.Methods WT1 gene level was detected by real time quantitative-polymerase chain reaction in acute myelogenous leukemia (AML) and in acute lymphocytic leukemia (ALL) patients.Then the expression levels of WT1 gene in different subtypes of AML were compared,and the correlation between gene expression and disease courses and prognosis were observed.Moreover,the relationship between disease courses and WT1 expression in patiens after receiving haemopoietic stem cell transplantation were analyzed.Results Among 66 cases,WT1 expression positive rate was 87.5 ％ (14/16) in AML and 76.0 ％ (38/50) in ALL.In AML,the expression level in M3 showed the lowest than that in any other subtypes (compared with M1,M2,M4,M5,P value was 0.040,0.007,0.006 and 0.01,respectively).The expression level of WT1 was closely correlated with leukemia disease courses.The expression level in complete remission (CR) group showed a significant lower expression level than that in non-remission group (P =0.018) and relapse group (P =0.003),and the re-increase of WT1 expression level could predict relapse as early as 1.5 months.Moreover,WT1 expression also showed an close relationship with prognosis of patients receiving haemopoietic stem cell transplantation.Patients whose WT1 was undetectable had a better prognosis than those with persistent expression,and increase again after becoming undetectable.Conclusion WT1 has a high expression level in AL,which can represent minimal residual disease.The expression level in M3 was lowest than that in different AML subtypes,and its expression level has a close correlation with clinical disease course and prognosis of AL.

Objective To study the clinic effect and safety of MEAD chemotherapy regimen for adult patients with relapsed or refractory acute lymphocyte leukemia. Methods Between July 2006 and July 2009,twenty-two adult patients with relapsed or refractory acute lymphocyte leukemia received MEAD regimen (mitoxantrone 6 mg/d dl-3 iv drip,cytarabine 100 mg/d dl-5 iv drip,etoposide 100 mg/d dl-5 iv drip,dexmethasone 10 mg/d dl-8 iv drip). Results The complete remission (CR) rate of adult patients with relapsed or refractory acute lymphocyte leukemia was 31.8 %,the partial remission(PR) rate was 22.7 % and the overall response (OR) rate 54.5 %. The cumulitive CR rate was 50.0 %,and the PR rate 40.9 % after two times MEAD chemotherapy regimen. The main adverse effect was different level of myelosuppression,and other toxicity of vital organ was mild. Conclusion MEAD regimen is effective and can be tolerated for adult patients with relapsed or refractory acute lymphocyte leukemia,and its side effect is mild.

Objective Ischemic stroke with elevated serum immunoglobulin E ( IgE) in some young patients is regarded as cerebral vasculitis clinically though without sufficient pathological evidence .This study was to investigate the characteristics of vascular lesions in these patients by temporal artery biopsy . Methods We performed histopathologic examinations on the temporal arteries of 32 young ischemic stroke patients with unknown etiology , 16 with normal and the other 16 with elevated serum IgE .We observed inflammatory cells infiltration and mast cells by HE staining and toluidine blue stai-ning respectively and determined the expressions of matrix metalloproteinase -9 (MMP-9), monocyte chemotaxis protein -1 (MCP-1) and serum IgE by immunohistochemistry . Results Compared with the patients with normal IgE , those of the elevated IgE group showed a significantly higher rate of inflammatory cells infiltration (12.5%vs 62.5%, P<0.01), with 1 case of focal necrosis and fi-brinous exudation in the adventitia in the latter group .The average optical density ( OD) of monocyte chemotaxis protein-1 ( MCP-1) in the temporal artery was also dramatically higher in the elevated IgE group than in the normal controls ([9.25 ±5.79] ×10 -5 vs [4.41 ±2.87] ×10 -5, P<0.01).The average OD of matrix metalloproteinase 9 (MMP-9) and intima-media thickness were both increased in the elevated IgE group ([32.79 ±21.38] ×10 -4 and [0.25 ±0.06] mm) but showed no statistically significant differ-ence from those in the normal IgE group ([25.23 ±12.78] ×10 -4 and [0.22 ±0.06] mm) (both P>0.05).Nor was any signifi-cant difference observed in the number of the mast cells between the normal and elevated IgE groups (2.8 ±1.5 vs 3.6 ±2.3, P>0.05). Conclusion The infiltration and necrosis of inflammatory cells and fibrin exudation in the temporal artery of the young pa-tient with elevated serum IgE are likely to be the manifestations of vasculitis , and MCP-1 may play a role in the pathogenesis of the disease.