OBJECTIVE: To establish the quality control system for the recombinant human GLP-1 analogue fusion protein. METHODS: The potency of the fusion protein was determined by luciferase reporter gene assay. The purity was analyzed by non-reduced SDS-PAGE and SEC-HPLC respectively. RP-HPLC was used for the peptide mapping. ELISA was used to analyze the identification of the final products. The molecular mass and peptide mass spectra were analyzed by LC-ESI-MS technique. Other control tests were performed according to the requirements in the Chinese Pharmacopoeia (Volume III, 2010 edition). RESULTS: Control tests were performed on three different lots of bulks and final products of recombinant GLP-1 analog fusion protein by the developed methods. The results showed that all the indexes met the requirements in the Guideline for Quality Control of Recombinant DNA Products for Human Use and Chinese Pharmacopoeia (Volume III, 2010 edition). The molecular weight of the recombinant human GLP-1 analogue fusion protein was 71 361.0, which was in conformity with the theoretical value. CONCLUSION: The developed methods and standard may assure the safety, effectiveness, and controllability of the recombinant human GLP-1 analogue fusion protein, which might be used for the routine quality control of products of the same kind.

The study aims to characterize and compare interferon reference standards from 5 manufacturers. By testing molecular mass and trypsin-digested peptide mass mapping, the amino acid sequence was verified and post-translational modifications such as disulfide bond were identified. Results show that the molecular mass and amino acid sequence were consistent with theory; the disulfide bonds of 4 lots of interferon were Cys1-Cys98/Cys29-Cys138, 1 lot was Cys29-Cys139/Cys86-Cys99; N-terminal "+Met", acetyl N-terminal and Met oxidation were identified in part of the sample. UPLC-MS can be used to characterize and compare interferon reference standards from different manufacturers.
Amino Acid Sequence ; Chromatography, High Pressure Liquid ; methods ; Interferons ; standards ; Mass Spectrometry ; methods ; Molecular Weight ; Oxidation-Reduction ; Peptide Mapping ; Protein Processing, Post-Translational ; Reference Standards

Objective To discuss effect on prevention of postpartum hemorrhage of caesarean section by us- ing calcium gluconate combined with oxytocin and misoprostol.Methods 385 cases of caesarean section were select- ed and randomized into O(Oxytocin) group and OM(Oxytocin+ Misoprostol) group and COM (Calcium gluconate+ Oxytocin+Misoprostol)group.Results The mean operative blood loss in O group and OM group and COM group were (300?50.24)ml,(220?30.83) ml,(150?45.52) ml.The amount of the mean operative blood loss of COM group was significantly lower than those of O group and OM group(P＜0.05).The amount of bleeding of 2 hours after delivery in O group and OM group and COM group were (400?45.52)ml,(260?60.43)mi and(210?50.54) ml.The amount of bleeding of COM group was significantly lower than those of O group and OM group (P＜0.05).Conclusion The prevention by used calcium gluconate combined with oxytocin and misoprostol is efficient in reducing the amount of postpartum hemorrhage of caesarean section.The operation of medicine is easy and safe and economic.

Background Arresting the overexpression of vascular endothelial growth factor (VEGF) will be a new approach to the inhibition of neovascularization.RNA interference (RNAi) can inhibit the expression of specific gene,and its application in eye has little interference to other gene expression.Objective This study was to investigate the effect of small interference RNA (siRNA) targeting VEGF on the expression of VEGF and retinal neovascularization in oxygen-induced retinopathy (OIR) model.Methods psi-HITM/enhanced green fluorescent protein (EGFP)/VEGF siRNA was designed and prepared in vitro.Mouse endothelioma (EOMA) were cultured in DMEM without antibiotic and divided into 5 groups.The cells were incubated in DMEM only in the blank control group;while 1 μl of LipofectamineTM 2000 + psi-HITM/EGFP,1 μl LipofectamineTM 2000 + 40,50 or 60 nmol/L of psi-HITM/EGFP/VEGF siRNA was added into DMEM in the negative control group and siRNA groups,respectively.The expression of VEGF mRNA and protein was detected by real time PCR (RT-PCR) and Western blot.The optimal effective concentration of VEGF siRNA was assessed.OIR models were established in 48 7-day-old C57BL/6J mice by raising them at an oxygen concentration of (75±2) ％ for 5 days and then to normal air.The mice were randomized into the model group,null vector group and VEGF siRNA group.1 μl of a mixture of psi-HITM/EGFP or VEGF siRNA (60 nmol/L) and LipofectamineTM 2000 was intravitreally injected,respectively,in the null vector group and VEGF siRNA group.The normal mice were used as the normal control group.Expression of VEGF mRNA and protein in the mouse retinas was detected by RT-PCR and Western blot,respectively,and FITC-dextran stretched retinal preparation was examined to evaluate the neovascularization,and retinal sections were examined to quantify the number of vascular endothelial cell nuclei extending beyond the internal limiting membrane (ILM).Results The in vitro transfection test showed that the expression of VEGF mRNA and protein in the EOMA cells was significantly different among the blank control group,negative control group and 40,50,60 nmol/L VEGF siRNA groups (F =148.890,P ＜ 0.001; F =306.960,P ＜ 0.001),and the expression of VEGF mRNA was lower in different concentrations of VEGF siRNA groups than that in the blank control group (t=73.950,119.890,156.480,all at P＜0.001).Also,the expression of VEGF protein was less in different concentrations of VEGF siRNA groups than that in the blank control group (t =15.452,23.344,42.119,all at P＜0.001).The optimal inhibitory concentration of VEGF siRNA was 60 nmol/L.In vivo study determined that compared to the model group and null vector group,the non-perfusion zones and neovascular net in the retina were decreased,and the number of vascular endothelial cell nuclei extending beyond the ILM was less in the VEGF siRNA group.The relative expression level of VEGF mRNA in the retinas was 1.23±0.18,4.02±0.16,3.98±0.19 and 1.98±0.12 in the normal control group,model group,null vector group and VEGF siRNA group,respectively,with a significant difference among them (F=369.780,P＜0.001),and the relative expression levels of VEGF mRNA in the model group and null vector group were higher than that in the normal control group (t=37.880,37.336,both P＜0.001),and the expression of VEGF mRNA in the VEGF siRNA group declined by 50.8％ (t=10.183,P＜0.001).The difference in the expression levels of VEGF protein also was assayed among the various groups (F=408.980,P＜0.001),and VEGF level in the retina was lowered by 68.0％ in the VEGF siRNA group compared to the model group (t =11.473,P＜0.001).However,VEGF level in the VEGF siRNA group remained at a high level in comparison with the normal control group (t =2.422,P＜0.001).Conclusions Intravitreal injection of VEGF siRNA can attenuate retinal neovascularization by effectively downregulate the expression VEGF mRNA and protein in the retina.

Biomarkers, Tumor ; metabolism ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; metabolism ; pathology ; Humans ; Lung Neoplasms ; drug therapy ; metabolism ; pathology ; Oncogene Proteins, Fusion ; chemistry ; metabolism ; Protein Kinase Inhibitors ; therapeutic use ; Pyrazoles ; therapeutic use ; Pyridines ; therapeutic use ; Pyrimidines ; therapeutic use ; Smoking

Aged ; Antigens, CD34 ; analysis ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Solitary Fibrous Tumors ; metabolism ; pathology

Objective To evaluate the effects of regular follow-up and systematic management in premature infants on early discovering of cerebral palsy and intervention efficacy in early phase. Methods Fifty premature infants with cerebral palsy recruited from early children development outpatient service, were classified as treatment group; 40 premature infants with cerebral palsy recruited from general outpatient service, were classified as control group. The infants of treatment group received systematic follow-up and intervention. while the infants of control group received no systematic management. Age and therapeutic efficacy were compared between the two groups. ResultsThe average age in the treatment group ( [4. 0 ± 1.2] months) was significantly younger than that of the control group ( [7.0 ± 1.4] months) ( P ＜ 0. 05 ). The mental development index ( MDI )and physical development index (PDI) in the treatment group was 91.8 ± 10. 2 and 90. 2 ± 11.2,respectively,which were significantly higher than that of the control group(80. 2 ± 11.7 for MDI and 79. 3 ± 10. 2 for PDI)( P ＜ 0. 05 ) . Conclusion Systematic management could help discover cerebral palsy in premature infants in time and could improve physical and mental development of these infants.

The genome of a new SV40 strain(SV-IMB)isolated from a rhesus monkey was completely sequenced and compared with other isolates.The results showed that the whole genome contains 5246bp,and the average identity of SV-IMB was 98.1% as compared to other SV40 isolates.Its regulatory region is composed of a complete enhancer and a defective enhancer.Amino acid changes occurred to some extent in both the large T antigen (T-Ag) and VP1 region.The findings demonstrate that the SV-IMB is a new SV40 isolate.

OBJECTIVETo assess the efficacy and safety of recombinant human thrombopoietin (rhTPO) on chemotherapy-induced thrombocytopenia in patients with solid tumor.

METHODSIn this randomized crossover self-controlled multi-center clinical trial, 154 patients with solid tumor were randomly divided into two groups (group A 77 cases and group B 77 cases). All patients were given the same two cycles of chemotherapy. In group A, the first cycle was treated cycle, in which patients were given rhTPO, while the second cycle was non-treated cycle as a control. In group B, the first cycle was non-treated cycle as a control, while the second cycle was treated cycle. RhTPO 1.0 microg/(kg x d) was administered subcutaneously 6-24 hours after chemotherapy for the longest 14 days. Laboratory tests included complete blood counts, urinalysis, serum chemistry, coagulant test, chest radiography, and electrocardiogram. Serum samples were screened for anti-rhTPO antibodies.

RESULTSIn both group A and group B, platelet decrease and duration had no significant difference between the treated cycle and non-treated cycle. Platelet count was higher in the treated cycle, than in the non-treated cycle: [minimal mean platelet count (64.4 +/- 45.4) x 10(9) cells/L and (52.4 +/- 30.9) x 10(9) cells/L (P=0.000), maximal mean platelet count (263.9 +/- 142.5) x 10(9) cells/L and (148.9 +/- 67.7) x 10(9) cells/L (P=0.000)]. Duration of thrombocytopenia was shorter in the treated cycle than in the non-treated cycle [days with platelet count < 50 x 10(9) cells/L, (2.5 +/- 3.9) and (3.7 +/- 5.7) (P=0.04); days with platelet count recovered > or = 75 x 10(9) cells/L, (10.3 +/- 8.7) and (14.0 +/- 8.9) (P=0.000), and days with platelet count recovered > or = 100 x 10(9) cells/L, (15.9 +/- 10.5) and (21.1 +/- 9.5) (P=0.000)]. The need for platelet transfusion was not significantly reduced in treated cycle. The effects of rhTPO on WBC, Hb, hepatic function, renal function, and coagulant function were not found. Transient low-titer non-neutralizing antibody was developed in one patient. Therapy with rhTPO was tolerated by all patients. Mild side effects were observed in individual patients, including fever, dizziness, and chill. Conclusion Administration of rhTPO after chemotherapy can significantly reduce the degree and duration of thrombocytopenia and promote platelet recovery. Therapy with rhTPO seems to be safe.

Adolescent ; Adult ; Aged ; Antineoplastic Agents ; adverse effects ; Cross-Over Studies ; Female ; Humans ; Lung Neoplasms ; blood ; drug therapy ; Male ; Middle Aged ; Neoplasms ; blood ; drug therapy ; Platelet Count ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Thrombocytopenia ; chemically induced ; drug therapy ; prevention & control ; Thrombopoietin ; therapeutic use