Purpose　 The aim is to establish the HPLC method for the determination of Fluoro-deoxyuridine in plasma.Methods　The Chromatography conditions include: Chromatography column: Nova-pak C18(3.9mm×150mm,4μm), mobile phase: 0.05mol/L sodium phosphate monobasic -methanol-water(0.5∶7∶92.5)， UV detection at 260nm， FUDR was extracted with ethyl acetate. Results　The average recoveries were 96.4%，96.5%，97.8% for concentration 0.23、1.67、20.0μg/ml (n=5).The corresponding reproducibility were RSD 1.61%, 1.98%, 3.17% respectwely for iner-day and RSD 3.56%, 1.90%, 2.63% for the intra-day(n=5). The FUDR concentration was linear with a correlation coefficient of 0.999　4 over the range of 0.099～20.0μg/ml. Conclusion　 The method was sensitive and accurate and suitable for pharmacokinetics and bioavailability study of FUDR.

Purpose　Both the working standard and corresponding sample(GRAN75)were first calibrated by WHO international standard for G-CSF.Methods　MTT method by NFS-60 cells was used. The results were calculated by (4,4)method.Results　Three batchs working standards were prepared,two batchs were freeze-dry and the prescription was same as WHO international standard for G-CSF,one batch was injection and the prescription was same as corresponding sample(GRAN75).The biological potency and the FL% of average potency were 3.062×106 、4.276×106IU/ampoule、1.635×107IU/ml and 5.529%、4.291%、4.244% for working standard, and 1.880×107IU/ml and 5.175% for corresponding sample,respectively.Conclusion　The working standard which calibrated could be used as working standard in the measurement of rhG-CSF biological activity.

Purpose　The aim is to study the influence factors of recombinant E.coli cell growth,product expression and purification,in order to raise the product rate of rhGM-CSF.Methods　Three pH media were planed to observe the effects of pH on E.coli cell growth and expression with the indexes of the cell density and expression level.The influences of various treatments on product rate were observed with the indexes of the volume and purity of inclusion bodies and the yield,purity,activity of product.Using SDS-PAGE method for analysis of purity.The product bioactivity was determined by using TF1 cell and MTT colorimetric method.Results　The cell density in the medium pH 7.0 was 2.22 g/L;the product expression rate was 23%.It was two times higher than other pH media.The inclusion body prepared by bacteria without freezing store was 2.5 times volume than freezing store.During renatura-tion PEG 4000 could raise 0.8 times active product.The optimum conditions used rhGM-CSF product rate could raise 8 times.Concluston　The product rate of rhGM-CSF expressed in E.coli was affected by many factors.The optimum condition for culture,expression and purification could raise quite a number of product rate.

Purpose　The aim is to investigate the hemostatic and protective effect of fibrin sealant(FS) on liver trauma in rats.Methods　On the surface of cracked wound(type Ⅰ) and resected wound(type Ⅱ)，spraying of FS or thrombin or direct suturation, was used receptively. Natural hemostasis was used as control.During postoperation the hemostatic time and the state of wound healing on 1st, 4th, 7th,14th, 50th day were observed.Results　Compared with the natural hemostasis,the hemostatical time of the FS group was shorter 86.0%(P＜0.01)in liver trauma type Ⅰ and 79.0%(P＜0.01)in liver trauma type Ⅱ.Compared with the thrombin group,the hemostatical time of the FS group was shorten 45.0%(P＜0.01)in liver trauma type Ⅰ and 84.0%(P＜0.05) in liver trauma type Ⅱ.The wound surface of FS group was healed faster than that of other groups.Conclusion　FS was an effective hemostatic and healing-promoter on liver trauma of rats.

Purpose　The aim is to investigate the effects of thymopeptide on LPS-induced release of nitric oxide (NO) from mouse peritoneal microphase in vitro.Methods　NO was detected by Griess reagent.Results　Thymopeptide 83.3～333.3 μg/ml reduced NO synthesis and secretion from mouse peritoneal microphase in vitro significantly.Conclusion　The effects of thymopeptide on LPS-induced release of NO from mouse peritoneal microphase may be one of mechanisms of its pharmacological effect.

Purpose　The aim is to develop a method for the analysis of micro-scale amino acids in biological samples.Methods　After having been extracted and dried,the residues were dissolved in a running buffer or a distilled water and acetonitrile.The samples were loaded into column for 5,25 or 50s at a hydrodynamic injection and were separated by capillary electrophoresis.The effect of different dissolvent on the sample stacking was observed.Results　A 100-fold improvement in the amount of material that can be injected into a capillary column without loss of resolution was shown.Conclusion　The marked effect of sample stacking was achieved by the using distilled water and acetonitrile(1∶1,v/v) as dissolvent.

Purpose　The aim is to study the conditions of preparation and regeneration of Gliocladium sp. (F) protoplast. Methods　 Different enzyme systems, enzymolysis time, osmotic pressure stabilizers were studied to investigate their influence on the productivity and regenerating rate of Gliocladiumsp. F protoplasts. The HPLC method was used to determine EP (6,22-diene-5,8-epidioxy ergosta-3-hydroxy) content.Results　The higher productivity of protoplasts was obtained when mycelia of strain F growing for 60 hours was digested at 28℃ for 4 hours by solution containing 2% cellulase and 2% helicase dissolved in 0.5 mol/L mannitol and the medium containing 0.5mol/L mannitol as osmotic pressure stabilizer would be suitable for protoplast regeneration. According to the EP productivity detected by HPLC, high positive rate of the regenerated strains growing in the medium containing 0.5mol/L mannitol could be got. Conclusion　The results will promote the research of strain F mutantgenesis and will be helpful for obtaining the strain more effectively biosythesizing compound EP.

Purpose　The aim is to establish fermentation process of new type recombinant human necrosis factor (nrhTNF)α.Methods　The optimized host cell,culture medium and induction time were deternined according to the growth and expression specificity of nrhTNF on test tube and flask shaker. Then fed-batch culture was carried out on 5 L automatic fermentor.Results　The expression of nrhTNF in fermentor could keep in a higher level of about 50% of total bacterial proteins when fermented to around A600nm 30，with a fermentation time of 12 h to 13 h.Conclusion　A short cycle,high expression and stabilized fermentation process was established.

Purpose　The aim is to evaluate the effects of rhIL-11 made in China on hematopoietic recovery following chemotherapy-induced thrombocytopenia in cynomolgus monkeys.Methods　Chemotherapy-induced myelosuppression of cynomolgus monkeys was made by china iv cyclophosphamide ( 30 mg/kg, qd ) for 5 days, then animals were divided 6 groups(n=4), by sc rhIL-11( 50, 100, 200 μg/kg), or Neumega (GI rhIL-11 control, 100 μg/kg) for 14 days, another normal control and model control. Blood was taken before chemothery and then at day 3, 6,9,12,15,18,21 for blood cell counts and platelet congregate test. Bone marrow was aspirated day 0, 12 and 21 for evaluation of the megakaryocyte ploidy distribution.Results　Recovery of blood platelets was accelerated and reached normal levels by day 12, and was higher than normal day 18, day 21 sc rhIL-11 in cynomolgus moneys. Blood platelet congregated rate were 71.4%～74.6% by day 21 and higher than model control . megakaryocytes in bone marrow increased.Conclusion　rhIL-11 could accelerate the recovery of peripheral blood platelets in monkeys. The function of increased platelets was normal. The results supported the clinical use of rhIL-11 as a platelet restorative agent to prevent severe thrombocytopenia following chemotherapy.

Purpose　The aim is to isolate and purify the erythropoietin(EPO)and thrombomodulin(TM) from human urine.Methods　Purifying and isolating human urine with means of filtration and concentration,ion-exchange chromatography, affinity chromatography and so on.Results　4.47 mg EPO and 9.92 mg TM were obtained from 2 000 kg human urine.Their molecular weights were(38.6±1.0)kD and (60±1.4)kD erespectively, the isoelectric point, 3.60±0.02 and 3.70±0.02, yield, 31.9% and 25.0%.The purified EPO and TM was shown to be highly homogeneous by capillary zone-electrophoresis(CZE),abundant Glu，Ala and Gly include on TM. Conclusion　The high purity EPO and TM was obtained.