OBJECTIVE:To establish a HPLC method for the simultaneous determination of quercitrin and isoquercetin in hypericum japonicum thunb.injection.METHODS:Merck Lichrospher RP 18 was used as chromatographic column and the mo-bile phase consisted of acetonitrile-(pH=3.0)buffer phosphate(18∶82)with flow rate at1ml/min,column temperature at25℃and detection wavelength at350nm.RESULTS:The linear range for quercitrin and isoquercetin were0.278?g～4.440?g(r=0.9991)and0.315?g～2.520?g(r=0.9991),respectively.The average recovery were98.60%(RSD=2.23%)and97.79%(RSD=1.74%),respectively.CONCLUSION:This method is accurate,reliable and applicable,and suitable for the quality control of hypericum japonicum thunb.injection.

This paper analyzes the present situation of intellectualization of hospital outpatient service building,points out its characteristics and disposition requirement,and proposes the design of intellectualized outpatient service.After rectifying outpatient procedure,our hospital designs and practices the new one,which provides humanism service for outpatient.

Purpose To evaluate apoptosis in renal tissue of diffuse proliferative lupus nephritis and therelationship between the existence of apoptosis cells in renal tissue and histopathological or clinical changes.Methods Apoptosis was detected by in situ nick-end labeling techniques (TUNEL) in renal biopsies from 25patients with type Ⅳ LN, 12 patients with IgAN, 4 patients with MsPGN, and 3 patients with APSGN. Normalrenal tissue obtained at nephrectorny for hypemephroma in 4 adults was used as control. In addition, proliferatingcells were identified by proliferating cell nuclear antigen(PCNA) in these patients. Results Compared to otherproliferative glomerulonephritis and control,the patients with lupus nephritis had less apoptosis cells, higher ratio ofPCNA+ cells/TdT+ cells/(P/T) in renal tissues;Ratio of P/T in glomeruli and tubulointerstitium correlated withthe chronicity index, r=0. 498 3(P = 0. 013 2), r = 0. 839 9(P＜ 0.001 ), r = 0. 661 4(P = 0. 003 3),respectively. Ratio of P/T in glomerulus and tubule had positive correlation with 24 hour urinary protein, r =0.855 4(P＜0.001),r=0.713 4(P=0. 001); negative correlation with Ccr, r = - 0. 488 0(P =0. 013 3)and r = - 0. 722 9(P = 0. 001), which in tubules positively correlated with Scr, r = 0. 410 7 (P = 0.041 4 ).Conclusions Apoptosis is insufficient in proliferative lupus nephritis. Intense proliferation without followingincrease in apoptosis may be related to chronic progressive renal histopatholcgical changes.

Laboratory of electron microscopy is representative of large instruments laboratories in medical research.And the operation and management in this kind of laboratory are different.Case of forty years of operation and management in the medical laboratory of electron microscopy,this paper analyzed and summarized its successful experiences in the research management system,personnel system and the aspect of equipment maintenance management.Provide a management reference for medical research institutions with the laboratory of similar large-scale instruments.

AIM: To investigate the protective effect of five natural anti-oxidants (schisandrin B, Sch B; silibinin, SIB; propyl gallate, PG; sodium ferulate, SF; total flavonoids of hippophase, TFH) on experimental oxidative injuries of lens. METHODS: All fresh transparent lenses of rabbit eyes except control group were bathed in Fenton reaction system in order to produce a model of oxidative damages of lens, meanwhile Sch B, SIB, PG, SF, TFH and pirenoxine sodium (PS) were added in the reaction system in different groups respectively. Lenses were incubated for 24 hours. Then total protein (TP), soluble protein (SP), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione (GSH), vitamin C (Vit C), total activities of anti-oxidation (TAO) and malondiaoldehyde (MDA) in homogenized lenses were measured to observe the effects of five anti-oxidants on above index. RESULTS: Five anti-oxidants increased the anti-oxidative index and decreased MDA in lens of oxidative damages in different levels, the effects are better than that of PS, especially in group SF and Sch B. CONCLUSION: The five natural anti-oxidants protected lens against experimental oxidative injuries very well. There are wide prospects to pursue effective anti-cataract drugs from natural anti-oxidants.

AIM: To investigate the effects of Co-SZ eye drop on galactose cataract in rats. METHODS: Based on folk remedy, SZ eye drop was made from leech, as a modified SZ eye drop, Co-SZ eye drop was enriched in Zinc and Vitamin C. In the present study, animal model of galactose cataract in SD rats was used. All animals were randomly divided into 3 groups : control group(using 0.9 % NaCl instead of SZ and Co SZ), SZ group and Co-SZ group. Lens opacities were examined dynamically in each groups via FS-3V slit-lamp microscope. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione (GSH) and soluble protein (SP) in the lenses were measured in 15 days. RESULTS: Both the Co-SZ and SZ eye drops could significantly delay and alleviate galactose cataract in rats, with better effect of Co-SZ than SZ eye drop. The antioxidant index indicated that SOD, GSH-Px, GSH in Co-SZ and SZ group were significantly higher than that in control group. Furthermore, SOD, GSH-Px in Co-SZ group were higher than that in SZ group significantly. CONCLUSION: Co-SZ eye drops could significantly delay and alleviate galactose cataract in rats, the effect is much better than SZ eye drops. The different effect between SZ and Co-SZ eye drops could be raised from the different content of Zinc, which is involved in anti-oxidation.

Objective To study the ultrastructure of neural stem cell neurosphere cultured in vitro. Methods Neural stem cell neurospheres from the rat brain were observed under transmission electron microscope or with stain of lanthanum nitrate, ruthenium red and tannic acid. Results The conjunction between the cells within neurosphere was loose and no tight junction was observed. Neural stem cells were proliferating lively. Some neural stem cells differentiated into cells with process and structure of axon, even showed the structures similar to myelin.Conclusion The ultrastructure of neural stem cell neurosphere cultured in vitro was revealed.

OBJECTIVE: We used the SD rat's bone marrow stromal cells (BMSCs) cultured in vitro to observe the effects of Bugu Mixture on the apoptosis and to explore the molecular biologic mechanism of the treatment of osteoporosis with Bugu Mixture. METHODS: BMSCs were separated from the bones of the extremities of SD rats in vitro. The morphologic changes, the apoptosis cell cycles, the mitochondrion membrane potential changes, and the Bcl-2 and Bax gene expression were observed, and the effects of Bugu Mixture on the course of cell apoptosis were evaluated. RESULTS: The earlier use of Bugu Mixture could decrease the cells blocked in G0/G1 phase, and promote their synthesis of DNA in S phase. The expression of Bcl-2 was higher in the Bugu Mixture group than that in the all-trans retinoic acid (ATRA) induced group, and the expression of Bax was lower in the Bugu Mixture group than that in the ATRA induced group. The mitochondrion membrane potential descended significantly in the Bugu Mixture group than that in the ATRA induced group. CONCLUSION: The mechanism of the treatment of osteoporosis with Bugu Mixture is that the earlier use of Bugu Mixture can decrease the amount of apoptostic cells induced by ATRA, thus promoting the cell mitosis and restraining the apoptosis. It can also act as a protector to Bcl-2 located on the mitochondrion membrane. By preventing the transferring of the Bax protein from cell-plasma to mitochondrion membrane, it takes the advantage of Bcl-2 in forming Bcl-2/Bax homodimer so as to prevent the opening of the permeability transition pore to avoid the changing of mitochondrion membrane potential and the destruction of biosynthesis caused by the mitochondrion release of apoptosis inducing factors and to reach the objective of restraining apoptosis.

Objective To express the novel fibronectin-binding protein Fba of group A streptococcus(GAS)and analyze its immunogenicity,so that to evaluate the immune responses to GAS infection.Methods fba gene was amplified by polymerase chain reaction(PCR)and confirmed by sequencing.Then it was cloned into pGEX4T-2 vector and Fba protein was expressed in E.coli BL21.The protein expression was identified by enzyme-linked immunosorbent assay(ELISA)and Westernblot.The sera from mice infected with GAS and anti-streptolysin-O positive patients were detected using microtiter plates coated with purified Fba protein as antigen.Afterward Balb/C mice were immunized with this purified protein and the sera were collected after the third immunization for the detection of IgG titer.Results It was confirmed by ELISA and Western blot that the recombinant Fba protein had a specific affinity with anti-Fba sera of rabbit.The anti-serum IgG titer of mice imrnunized with Fba protein was up to 1:4800.Conclusions GAS infection or Fba protein immunization are able to induce high serum titer of anti-Fba which could react specifically with the recombinant Fba protein.It indicates that Fba protein has good immunogenicity and antigenicity.So Fba protein could be a GAS candidate vaccine and an important tool to detect anti-GAS titer in GAS infected patients.

Objective:To observe cellular,humoral and mucosal immune responses induced by DNA-or protein-based of FimH of UPEC type 1.Methods:After mice were immunized respectively with recombinant plasmid pcDNA3.1/fimH or pcDNA3.1/fimC,and the combinant FimH and FimC protein,the anti-FimH protein IgG of sera and SIgA in bladders were detected by ELISA.The lymphocyte phenotypes of CD3,CD4 and CD8 were analyzed by FCM.Results:The titers of IgG in sera and SIgA in the bladders were all low in the group immunized by recombinant FimH plamid,but the percentage of CD4+T cells in spleen was high,which revealed that recombinant FimH plamid was able to trigger better cellular immune response.The titers of IgG were very high in the group immunized by FimH protein,which suggested that the FimH protein was able to trigger better humoral immune response,but SIgA in the bladders was not detectable.Conclusion:The DNA for FimH can induce humoral,mucosal and cellular immune response.FimH protein can only induce humoral immune response.FimC protein is able to enhance the immunogenicity of FimH protein.