Objective: To study the effect of eliminating pyrogen with ultrafiltration on the quatity of Chuanshentong Injection. Methods: The sample solution was filtered with the ultrafiltration membranes of molecular weight 6000 and 20000. Then,the results of eliminating pyrogens and the examination results of quality of sample solution before and after ultrafiltration were compared.Results: The results of elminating pyrogens is satisfactory. But there are larger changes in many items of quality markers after ultrafiltration. Conclusion: Therefore, the ultrafiltration should be used with caution in eliminating pyrogens of injection of Chinese medicinals.

Objective To investigate the effect of MT-1 and MT-2 in liver damage in chronic inorganic arsenate exposure mice and to explore the mechanism of arsenic-induced liver damage. Methods The male Kunming mice were randomly divided into control and exposed groups. The control group was given ordinary feed and tap water. The exposed group was given ordinary feed and 300 mg/L of sodium arsenite solution by drinking water. After 10 months of treatment, the activity of serum alanine aminotransferase (ALT), aspartate aminotransferase(AST) and globulin (Glb) content were determined. The total RNA was extracted by the TRIzol-phenol-chlorofor-method from the liver tissue. The quantity of the RNA was determined by spectrophotometry and its purity was judged at a ratio of A260/A280. Then real-time PCR(RT-PCR) was used to measure the mRNA expression of MT-1 and MT-2. Results Compared with the control group, serum ALT, AST activity and Glb content were higher and the MT-1 and MT-2 mRNA content was lower in the exposed group (P

Objective To investigate changes of the visual P300 topography mapping in patients with closed craniocerebral trauma.Methods The visual P300 topography mapping was recorded from 103 patients with closed craniocerebral trauma and 66 normal subjects with a medicide-03E brain evoked potential instrument. Results The P300 latency in patient group was significantly prolonged as compared with the control group ( P

ObjectiveTo enrich and quantitatively analyze peripheral circulative carcinoma cells by magnetic activated cell sorting (MACS) in colorectal cancer patients.MethodsBlood samples, preoperatively collected from 21 patients with colorectal cancer and 9 healthy volunteers, were labeled by anti-cytokeratin (CK) with magnetic microbeads, and passed through magnetic columns; CK +carcinoma cells were separated from the samples, and quantified by flow cytometry.ResultsCK +CD45 -cells could not be detected from the samples without MACS; in the sample undergoing MACS, the concentrations of CK +CD45 -cells were significantly higher in patient group than in control group(P

Objective:To study inhibited tecomerase activity of HL-60 cells in human cancer cell by Icarrin(ICA) and its mechanism.Methods:MTT assay,NBT assay, TRAP-PCR,RT-PCR assay,Flow Cytometry.Results:ICA inhibited telomerase activity in HL-60 cells significantly.It was negative correlation between telomerase activity and the expression ratio of CD11b antigen in HL-60 cells.It can induced HL-60 cells to differentiate into mature granulocytes.It can changed cell cycle distribution of HL-60 cells,which was reflected by the accumulation of the vast majority of cells in the G0/G1phase and the loss of cells in the S phase. It can downregulated cell proliferation related gene c-myc;upregulated p21 gene protein and mRNA expression.Conclusion:This study proved mechanism of ICA on inhibited telomerase activity from gene-protein-effect of cell level in HL-60 cells.

Objective To observe the expression of c-fos mRNA and protein in fluoride treated mouse fibroblast (FB) and osteoblast (OB) and to further explore the effects of c-fos in the osteogenic action of FB. Methods Mouse FB and OB were divided into control group and six fluoride groups (0, 0.0001, 0.0010, 0.1000, 1.0000, 10.0000,20.0000 mg/L F-), and the levels of c-fos protein at 2,4,24,48,72 h and c-fos mRNA at 48 h were measured by using ELISA and RT-PCR methods. Results Compared with the control group, fluoride increased the content of c-fos protein obviously in all FB group(P<0.01); and it is increased in 0.0001,0.0010 mg/L groups at 48 h (0.73±0.04, 0.64±0.14) and 0.0001 mg/L group at 72 h(0.70±0.17) in OB compared with the control group (0.32±0.04,0.27±0.05, P<0.01). Compared with the control group (0.95±0.11), RT-PCR revealed an increasing tendency of the expression of c-fos mRNA at 48 h in FB (1.06±0.16, 1.06±0.12,1.12±0.16,1.04±0.15,1.04±0.10,1.15±0.29), but the difference was not statistically significant(P>0.05); however, a statistically significant difference(P<0.01) of c-fos mRNA in 20.0000 mg/L group(1.40±0.17) in O B was found compared with the control group (1.06±0.06). Conclusion The higher expression of c-fos mRNA and protein in FB induced by fluoride may play an important role in the transformation of osteoblastic phenotype as well as increase the osteogenesis ability in FB.

Purpose The aim of this study was to reveal the role of SGT,the small glutamine-rich tetratricopeptide repeat (TPR)-containing protein,in the developing mouse brain through examining the expression profile of SGT during the development stage. Methods In this study, quantitative RT-RCR and Western blot were applied to investigate the expression of SGT mRNA and protein in the mouse whole brain. Western bolt was also used to detect the expression of SGT in the hippocampus, cerebral cortex, striatum and cerebellum. Immunohistochemical analysis on postnatal and adult mouse brain was performed to examine the subcellular localization of SGT. Results Our data showed that the levels of SGT mRNA and protein in the mouse whole brain were both high during the postnatal stage and declined in the adult. Regional expression of SGT protein in the hippocampus, cerebral cortex, striatum and cerebellum showed a similar expression profile.Immunohistochemical analysis found that in the P14 mouse brain, SGT was abundant in all the CA regions of hippocampus as well as most regions of cerebral cortex and striatum. In the cerebellum, SGT was mainly distributed in Purkinje cells. In the sections of the adult mouse brain, faint expression was observed in the regions mentioned above. Conclusions Our findings firstly exhibit the expression pattern of SGT in the mouse brain development,which might shed new light on further functional analysis of SGT in the central nervous system.

Objective To clone human asparagine synthetase(ASNS)gene,express the MS2- ASNS fusion protein through gene engineering and use the purified target protein to immune BALB/C mice, prepare and identify the monoclonal antibody(McAb),which forms the base for studying mechanism of L- asparaginase used as salvage regimen in midline NK/T cell lymphoma nasal type.Methods ASNS gene fragment was amplified by RT-PCR from HepG2 cell line and constructed into prokaryocytic expression vector.Fusion-protein of MS2-ASNS was expressed and used for immunizing BALB/C mice to prepare McAbs against ASNS.Identified the McAb and detected the expression of ASNS in tumor.Results Part of the ASNS reading frame(NCBI,M27396:179-1 420 bp)was cloned and product length of RT-PCR was 1 263 bp.Molecular weight of MS2-ASNS was about 54 700 Da.Two strains of hybridoma secreting ASNS McAbs were obtained.The subtype of the ASNS McAb was IgG2a.Western-blot showed that the McAbs could specifically react with MS2-ASNS fusion protein and ASNS protein in tumor cell lines.ASNS expression was detected by immunocytochemistry and Immunohistochemisrry.Conclusion We have cloned human ASNS gene,obtained the anti-ASNS McAb and examined the expression of ASNS in tumor.

Objective:To study the role and mechanism of c myc ASODNS on cell expression of surface antigen of the tumor cells and susceptibility of target cells to immune cells attack.Methods:c myc mRNA was examined by RT PCR.MTT assay was usd to explore the effects of c myc ASODNS on PG cell sentisentity to lysis by CD 3AK effector cells.The expression of HLA ABC,ICAM 1,c myc protein was examined by flow cytometry.Results:When PG cells were treated with ASODNS(1 ?mol/L) there was a markably reduction of expression of c myc protein.Expression ratio of HLA ABC and ICAM 1 surface antigen expression ratio of PG cells were enhanced from 68.44%,38.44% to 83.16% and 42.09% respectively( P

Objective To explore the biological characteristics and karyotype of bone marrow-derived mesenchymal stem cells(MSCs)in patients with systemic lupus erythematosus(SLE)and hematopoietic sup- port of MSCs.Methods MSCs were isolated from bone marrow of 11 SLE patients and 6 healthy controls by density centrifugation and adhesive culture in vitro.The surface markers were detected by flow cytometry (FCM).The morphological changes of MSCs were observed in primary and passage cultures.The growth curves were assayed.The karyotype of MSCs was detected by blocking cellular mitosis with colchicines.The MSCs from SLE patients and healthy controls were infused to ICR mice after high-dose chemotherapy.The changes of peripheral blood counts of the mice were recorded.Results Approximately(6～9)?10~9 MSCs from SLE were obtained after 5 passages and their growth was slower than normal controls(P＜0.01).Both groups were positive for CD29,CD44 and CD105,and negative for CD14,CD34,CD45 and HLA-DR.MSCs from SLE had a normal karyotype.MSCs infusions of the two groups were accompanied by no adverse event and the recovery of white blood cell,hemoglobin and platelet count was quicker when compared with the controls(P＜0.05).Conclusion MSCs from SLE have demonstrated abnormalities in expansion in vitro.MSCs from SLE have a normal karyotype.Ex vivo MSCs infusion from SLE patients can support hematopoiesis as normal MSCs.