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Journal of Jiangsu University(Medicine Edition)

	2013 Vol.23 Issue.3 Published 2013-05-30

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		2013 Vol. 23 (3): 0- [Abstract] ( 710 ) [HTML 1KB] [ PDF 322KB] ( 1354 )

	185	Osteogenic differentiation of primary cultured human adiposederived mesenchymal stem cells accompanying adipogenic differentiation in vitro
		MIN Min-1, ZHANG Xue-Jing-2, MA Hong-1, XU Hui-1, LI Yu-Mei-1
		Objective: To establish the isolation and culture method of human adiposederived mesenchymal stem cells (hADSCs) derived from human adipose in vitro, so as to explore their morphology, identify cell surface markers, observe biological properties, and discuss the possible causes and mechanism of the phenomenon of hADSCs′ osteogenic differentiation accompanying with synthesis of triglycerides. Methods: Human adipose tissue were obtained from abdominal operation. The hADSCs were isolated from human adipose tissue by 0.15% collagenase digesting. The cells were applied to do the experiments : MTT method, flow cytometry, immunofluorescence. Its differentiation potential was proved by osteogenic and adipogenic differentiation. The osteogenic and adipogenic related genes: PPARγ2, osteopontin expression were detected by realtime fluorescent quantitative PCR technique. Results: After isolation and culture, we obtained a large amount of hADSCs, which grew like swirls. MTT revealed high capability for and proliferation. The flow cytometry showed CD29+, CD31-, CD34-, CD45-, CD73+, CD105+, CD166+, HLADR-, which fit the results of immuno fluorescence. Moreover, these cells could be functionally induced into adipocytes and osteoblasts in the presence of appropriate conditioned media.During osteogenic differentiation, we found it accompanying with the synthesis of triglycerides.RT-PCR results proved that during the differention process, osteogenic and adipogenic related genes began to be expressed gradually, which had statistically significant(P<0.01). Conclusion: Highly efficient isolation and cultivation methods for hADSCs have been developed. They are a kind of mesenchymal cells with great application prospect, which characterized with adherent growth, high proliferation, stem cell phenotype and multipotent differentiation. During vitro osteogenic differentiation, the triglyceride formation has a certain role in promoting osteogenesis.
		2013 Vol. 23 (3): 185- [Abstract] ( 1471 ) [HTML 1KB] [ PDF 5671KB] ( 1953 )

	191	Potential mechanism and inhibitory effects of silver nanoparticles on parainfluenza virus type 3
		YIN Jian-Jian, Li-Xiu-Jing, Zheng-Cong-Long
		Objective: To explore the inhibitory effects and the potential mechanism of silver nanoparticles against parainfluenza virus type 3(PIV3). Methods: Cell culture, MTT test assay and immune fluorescence assay were applied to evaluate the antiPIV3 activities of silver nanoparticles in preventing, treating and directly inactivating. The neuraminidase activity inhibition test was used to study the inhibitory effect of silver nanoparticles on PIV3 neuraminidase activity; destructive effect of silver nanoparticles on PIV3 was observed by transmission electron microscope (TEM); murine experiment was performed to observe the pathological changes of mouse lung tissues in vivo. Results: The survival rates of canine kidney cells(MDCK) were 93.05%，90.32% and 94.81%, respectively, under three different administration(treatment to medicine group, preventive medication group, direct inactivating group) of silver nanoparticles by MTT assay, while the cell livability in control group was 25.50%, the differences were statistically significant (P<0.01). MDCK cells infected with PIV3 had more specific yellowishgreen immunofluorescence compared with the cells treated with silver nanoparticles in virucidal, antiviral and inactivating ways. The inhibition rate of neuraminidase activity on silver nanoparticles treatment group was higher than 80%, while the PIV3 control group and the solvent group were lower than 20%. The results of TEM showed that silver nanoparticles had an obvious destructive effect on PIV3 in a timedependence. Compared with the normal lung tissues, the pathological changes in the mice treated with the silver nanoparticles were similar or less severe, while the lung tissues from the virus model group presented with edema, inflammatory cell infiltrating and morphological structure disappearing. Conclusion: Silver nanoparticles had remarkably inhibitory effects on PIV3 and the possible mechanisms may be associated with the neuraminidase inhibition and virion damage by silver nanoparticles.
		2013 Vol. 23 (3): 191- [Abstract] ( 1753 ) [HTML 1KB] [ PDF 5220KB] ( 1834 )

	197	Expression and effect of miR375 on synthesis of collagen Ⅰ in mice with diabetic cardiomyopathy
		HUANG Yan, ZHANG Ying-Yu, WANG Hao, ZHOU Yan-Fang, ZHANG Guo-Hui, RUI Tao
		Objective:To investigate the effect of miR375 on cardiac fibrosis in diabetic cardiomyopathy mice.Methods: Cardiac fibroblasts isolated from hearts of C57BL/6 mice were cultured in DMEM medium(low glucose concentration).The purity of cardiac fibroblasts were evaluated by immunohistochemical staining.The second and third generation cardiac fibroblasts were randomly divided into control group(5 mmol/L glucose)and high glucose group(25 mmol/L glucose). The cardiac fibroblasts of high glucose groups were treated by high glucose for different times(6，12，24 h).The different expression of miR375 in cardiac fibroblasts was detected by realtime quantitative PCR.miR375 inhibitor was transiently transfected into cardiac fibroblasts by lipofectamineTM 2000 reagent； transfection efficiency was detected through realtime quantitative PCR. The expression of collagen Ⅰin cardiac fibroblasts was evaluated by immunohistochemical staining.Results: The purity of the second generation of cardiac fibroblasts was above 90%.During the time point 6 to 24 h cardiac fibroblasts treated by high glucose，the expression of miR375 showed an increase tendency；in high glucose and miR375 inhibitor treated group，the expression of collagen Ⅰdecreased significantly. Conclusion: MiR375 might play an important role in diabetic myocardial fibrosis.
		2013 Vol. 23 (3): 197- [Abstract] ( 1422 ) [HTML 1KB] [ PDF 3728KB] ( 1571 )

	201	Effect of macrophages on the migration of mouse bone marrow mesenchymal stem cells in inflammation
		GAO Shuo, CAI Meng-Jie, MAO Fei, YANG Ting-Ting, ZHANG Jie, ZHANG Ling, ZHAO Lu-Lu
		Objective: To investigate the effect of macrophage cell line RAW264.7 on the migration and expression of TGF-β1, IL-6 and MCP-1 gene level of mouse bone marrow derived mesenchymal stem cells(MSCs) under the stimulation of lipopolysaccharide(LPS). Methods： Adherent method was used to isolate mouse bone marrow mesenchymal stem cells. MSCs were co-cultured with RAW264.7 through transwell assay. Co-culture experiments were divided into four groups: culture medium control, RAW264.7 cells, LPS and RAW264.7 cells stimulated by LPS. Cell transwell migration assay were performed to detect the number of migrated MSCs under the four different conditions. At the same time, qRT-PCR was performed for the quantification of TGF-β1, IL-6 and MCP-1genes. Results： RAW264.7 expressed high level of TNF-α, IL-6, IL-1β and MCP-1 gene by LPS stimulation. In the presence of RAW264.7, the migrated MSCs numbers were higher than controls(P<0.01), while RAW264.7 cells remarkably increased the migratory ability of MSCs under the stimulation of LPS(P<0.01). Meanwhile, the expression of TGF-β1, IL-6 and MCP-1 genes were enhanced by co-culture with RAW264.7 in the presence of LPS(P<0.05). Conclusion： Macrophage cell line RAW264.7 enhanced the migratory ability and exchanged the cytokine expression of MSCs under the stimulation of LPS, which provided mechanistic insights into the recruitment of MSCs in inflammation.
		2013 Vol. 23 (3): 201- [Abstract] ( 1607 ) [HTML 1KB] [ PDF 4575KB] ( 1589 )

	207	Prokaryotic expression and identification of rhoptry protein 18 from Toxoplasma gondii
		JU Ai-Ping, WU Liang, SHEN Jin, LI Li, JIANG Xu-Gan, CHEN Sheng-Xia
		Objective:To clone and express the rhoptry protein 18(ROP18) from tachyzoites of the RH strain of T.gondii and establish methods to determine and locate the expression of ROP18 in T.gondii. Methods: The gene fragment of ROP18 was amplified by PCR from the cDNA of T.gondii RH strain and cloned into pET-28a(+) vector. The recombinant ROP18 was expressed in E.coil Rosetta strain and purified by cutting the gel slices with KCl stain. AntiROP18 polyclonal antibody was produced in rabbit. The ROP18 in T.gondii was determined and located by Western blotting and indirect immunofluorescence assay (IFA) respectively. Results: The recombinant and its polyclonal antibody were obtained. The specific band of ROP18 in T.gondii was detected using polyclonal antibody by Western blotting. The ROP18 was distributed in the rhoptry of T.gondii by IFA. Conclusion: Western blotting and IFA could detect and locate ROP18 expression using anti-ROP18 polyclonal antibody. It will facilitate the study of the biological functions of ROPs.
		2013 Vol. 23 (3): 207- [Abstract] ( 1185 ) [HTML 1KB] [ PDF 3779KB] ( 1492 )

	212	Neurotoxic effect of aconitine based on the midbrain dopaminergic neurons cell culture
		YAN Yan-1, YANG Mao-2, Wolfdieter-Rausch3 , WANG Xi-Jun-1
		Objective: To observe toxicological effects of aconitine on midbrain dopaminergic neurons and to understand and prevent neurotoxic side effects from aconitine and its derivatives. Methods: Fetal rat brain was taken out from the 14 days pregnant mice and dopaminergic neurons were isolated to culture in 5％ CO2, 37 ℃ and 100％ humidity conditions. Different concentrations of aconitine (0.1, 0.5, 1, 5, 10, 50 and 100 μmol/L) were added on 10th day in vitro and treated for 48 hours. Immunohistochemical staining was done on 12th day and the number of cells, dendritic length and the number of neuron axons were selected as observed indicators, and to make the analysis. Results: Fortyeight hours later, different concentrations of aconitine(0.5-100 μmol/L) had strong inhibition on growth of neuron cells, which showed a significant concentrationdependent neurotoxicity. While the high concentrations of aconitine (10～100 μmol/L) had stronger influence on neuron cells than the low concentrations of aconitine (0.5～5 μmol/L). Conclusion: The neurotoxicity effects of aconitine on midbrain dopaminergic neurons in vitro experiments were confirmed for the first time.
		2013 Vol. 23 (3): 212- [Abstract] ( 1271 ) [HTML 1KB] [ PDF 3170KB] ( 1353 )

	216	Expression and role of S100B on the injury of cardiomyocytes induced by doxorubicin
		HAN Yao-Hui, PAN Hao, MAO Jun-Qian, ZHOU Yan-Fang, ZHANG Ying-Yu, WANG Hao, BAO Yong-Hui, ZHAO Long, LIU Ling-Ling, ZHANG Guo-Hui
		Objective: To investigate the expression and the role of S100B on the injury of cardiomyocytes induced by doxorubicin. Methods: The cardiomyocytes from 1d SD rats were cultured in DMEMF12 medium and then randomly divided into 4 groups: control group(CON), doxorubicin damnified group(DOX), doxorubicin+negatived small interfering RNA control group(DNC), and doxorubicin+S100B small interfering RNA group(DSB). S100B small interfering RNA was mediated by lipofectamineRNiMAX to transfect cardiomyocytes. The cardiomyocytes were treated with doxorubicin(2 μmol/L)for 2 h at 48 h posttransfection. The survival rate of cardiomyocytes was evaluated by MTT. The apoptosis rate was determined by flow cytometry(FCM). The expression of S100B was detected by Western blot. Results: MTT assay showed that there was no significant difference in the survival rate of cardiomyocytes between DOX group and DNC group(P>0.05). Compared with DOX group, the survival rate of cardiomyocytes in the DSB group was increased(P<0.05); Flow cytometry showed that there was no significant difference in the apoptosis rate of cardiomyocytes between DOX group and DNC group(P>0.05). Compared with DOX group, the apoptosis rate of cardiomyocytes in the DSB group was descended(P<0.01); Western blot showed that there was no significant difference in the expression of S100B between DOX group and DNC group(P>0.05), Compared with DOX group, the expression of S100B in the DSB group was descended(P<0.01). Conclusion: The expression of S100B were significantly increased on the injury of cardiomyocytes induced by doxorubicin and might promote the apoptosis.
		2013 Vol. 23 (3): 216- [Abstract] ( 1425 ) [HTML 1KB] [ PDF 3911KB] ( 1458 )

	220	Establishment of a mouse model of human tumor based on principle of oral tolerance
		LIU Xia-1, CHENG Jing-1, FAN Xing-Wen-1, ZHOU Xiao-Bing-2, ZHOU Dan-1, WU Wei-Jiang-1, HUA Ye-3, SHAO Qi-Xiang-1
		Objective: To establish a mouse model of human tumorspecific immune tolerance based on principle of oral tolerance. Methods: Ninety mice were randomly divided into three groups: blank control group, irrelevant tumor control group and experimental tumor model group, thirty in each group. The mice of three groups were received intragastric administration of normal saline, human ovarian cancer cell line 3AO and human hepatoma cell line HepG2 cells for 7 days, respectively. Then, all mice received subcutaneous injection of HepG2 cells in the back skin. The time course of the tumor incidence were recorded and the size of tumor were determined by slide caliper. The histological examination of sections of tumor tissues were evaluated by optical microscope. Results: HepG2 cells in mice from experimental tumor model group survived and developed into tumor after subcutaneous injection of HepG2 cells. The tumor incidence was 90％ (27/30). There was no tumor formation in the mice of blank control group and irrelevant tumor control group. Obviously pathologic changes were found in tumor tissue sections with HE staining, while some debris of death tumor cells were found in mice from control group and irrelevant tumor control group. Conclusion: The mouse model of human tumor was established successfully by intragastric administration of HepG2 cells through induction of specific immune tolerance.
		2013 Vol. 23 (3): 220- [Abstract] ( 1785 ) [HTML 1KB] [ PDF 3917KB] ( 1879 )

	224	Establishment of techniques for generation in vitro and isolation ex vivo for human primary mast cells
		DI Rong-Rong-1, 2 , JIANG Jin-Feng-2, ZHANG Ji-3, DING Yong-Bin-3, WANG Jian-Hua-2, WEI Ji-Fu-1
		Objective: To establish the technique for generation or isolation of human primary mast cells,and for further studying their roles in regulating host immunity and elucidating immunopathogenesis. Methods: CD34+hemopoietic stem cells were cultured in presence of stem cell factor, IL6, and IL3 to generate primary mast cells.Mast cells allocated in human intestinal tissues were isolated through digestion, density gradient centrifugation and purification with specific antibodiescoated magnetic beads. Cell phenotype was monitored by immuostaining of surface markers of CD117 and FcεRⅠ and intracellular tryptase, or cells were identified with Toluidine blue staining. Results: Both of the CD34+cellderived and intestinal mast cells expressed CD117 and FcεRⅠ， the specific markers for mast cells, and contained intracellular tryptase. Conclusion: Both methods could provide sufficient and purified primary mast cells for further study in regulating host immune response and defence against pathogenic microorganism.
		2013 Vol. 23 (3): 224- [Abstract] ( 1425 ) [HTML 1KB] [ PDF 4761KB] ( 2503 )

	229	Influence of aerobic exercise on cell apoptosis and expressions of Bcl-2 and Bax protein in skeletal muscle from aging model rats
		LIU Ning-Ning-1, LI Ming-Xue-2, JIN Wen-1
		Objective: To investigate the effect of aerobic exercise on cell apoptosis and expressions of Bcl-2 and Bax protein in skeletal muscle from aging model rats. Methods: The aging model rats were randomly divided into control group and two exercise groups which include 6 times/wk exercise group and 3 times／wk exercise group. Exercise groups were trained by swimming for 12 weeks, 90 min each time. All rats were killed after 48 hours of the last training session. Cell apoptosis of skeletal muscle was detected by using TUNEL staining. The expressions of Bcl-2 and Bax in skeletal muscle were quantified by western blotting. Results: Compared with control group, the apoptotic index(AI) of skeletal muscle and expression of Bax protein in exercise groups were obviously decreased (both P<0.01), while the expression of Bcl-2 protein and the ratio of Bcl-2/Bax were markedly increased (both P<0.01). Compared with 6 times／wk group, the AI of skeletal muscle and expression of Bax protein in 3 times/wk group were significantly decreased(P<0.05), whereas the expression of Bcl2 protein and the ratio of Bcl-2/Bax were greatly increased(P<0.05). Conclusion: The ratio of Bcl-2/Bax could be increased by aerobic exercise. And the excessive apoptosis of skeletal muscle of aged rats could be reduced by aerobic exercise.The effect of everyotherday aerobic exercise was better than everyday aerobic exercise. It was beneficial to inhibit cell apoptosis of skeletal muscle.
		2013 Vol. 23 (3): 229- [Abstract] ( 1388 ) [HTML 1KB] [ PDF 3717KB] ( 1682 )

	233	Inhibitory effect of ginkgols on SMMC-7721 liver cancer cells in vitro and liver cancer H22bearing mice in vivo
		WANG Yun-Fei-1, YANG Xiao-Ming-1, Li-Yue-Ying-2, Li-Jie-1, Huang-Bing-Zhong-3, Guo-Chan-Yuan-4, Yi-Nan-1, Xing-Chao-Hui-1
		Objective: To study the inhibitory effects of ginkgols on SMMC-7721 liver cancer cells in vitro, and on tumor growth of H22bearing mice in vivo. Methods: The inhibitory effect of ginkgol on SMMC-7721 cells was evaluated by the survival rate of SMMC-7721 cells using MTT assay; then half maximal inhibitory concentration (IC50) was calculated. The solid tumor model of liver cancer H22bearing mice was established, and all mice were randomly divided into model control group, cyclophosphamide(CTX) group, four ginkgol groups(20, 40, 80, 160 mg/kg), eight in each group. All mice in the groups received intraperitoneal injeciton of normal saline(0.2 mL each), CTX(2 mg/kg body weight), ginkgol 20, 40, 80, 160 mg/kg, respectively, for 7d. At 24 hours after the final injection, blood sample was collected by eye bleed; samples of tumor tissues, spleen and thymus were harvested after mice were killed. The inhibition rate of the antitumor, spleen index and thymus index were calculated. The levels of lymphocytes in peripheral blood were measured. Results: MTT assay showed that ginkgols inhibited the proliferation of SMMC-7721 cells with half maximal inhibitory concentration (IC50) values of (15.33±1.19), (12.20±1.08) and (10.88±1.03) mg/L for 24, 48, and 72 hours, respectively. The inhibition rate of 20, 40, 80 and 160 mg/kg group were 8.96%、39.81%、48.51% and 28.36%, respectively. Except for 40 mg/kg group, there were significant differences in inhibition rate between the other ginkgols groups and model control group(both P＜0.05). The levels of lymphocytes in ginkgols groups from peripheral blood were significantly increased, compared with CTX group(P<0.01). Conclusion: Ginkgols showed markedly inhibitory effects on cell proliferation of human hepatic carcinoma cell lines SMMC7721 in vitro, and on tumor growth of H22bearing mice, which could be associated with the regulation of immune function.
		2013 Vol. 23 (3): 233- [Abstract] ( 1539 ) [HTML 1KB] [ PDF 3699KB] ( 1555 )

	238	Preparation and identification of monoclonal antibody against Helicobacter pylori CagL protein
		ZHU Hong, Chen-Cheng, Ling-Feng, Shao-Chen, Yu-Min, Shao-Shi-He
		Objective: To prepare and characterize the monoclonal antibody(mAb) against Helicobacter pylori (H.pylori) CagL protein. Methods: The CagL fusion protein was induced to express by IPTG in E.coli BL21(DE3) which contains recombinant expression vector pET-28a (+) and then purified by Ni^2＋-NTA resin. Purified CagL fusion protein was used to immunize 6week BALB/c mice, and then the spleen cells were fused with myeloma SP2/0 cells; the positive hybridoma cell lines were obtained by means of several rounds of subclone. Immunoglobulin isotype of mAb was identified by ELISA method. The titer of mAbs in cell culture supernatant and ascites, as well as the relative affinity of these antibodies, were measured by indirect ELISA method. The antigenic epitopes of mAbs were evaluated with ELISA additivity test. The specificity for antibody was determined by Western blotting. The stability of hybridoma cell lines specifically secreting mAbs was tested by cryopreservation and resuscitation. Results: Three hybridoma cell lines which could secrete stably mAbs against CagL were successfully obtained. The isotype of two mAbs was classified as IgM, another IgG2a, and the light chain subset of three mAbs was classified as type κ. The titers of three mAbs in cell culture supernatant were 1∶64～1∶128, while those in ascites reached more than 1∶16 000. The affinities of mAbs were found to be high. Three different antigenic epitopes might be recognized by the mAbs. Western blotting analysis showed that all three mAbs could bind to the CagL fusion protein; however, only two of them had specific reaction with the whole protein extracted from H.pylori strains. After cryopreservation, resuscitation and subculture, the titers of mAbs secreted by three hybridoma cell lines were still high. Conclusion: The monoclonal antibody against CagL protein was successfully prepared, which would play a significant role for the further study in the mechanisms of CagL as well as H.pylori type Ⅳ secretion system.
		2013 Vol. 23 (3): 238- [Abstract] ( 2507 ) [HTML 1KB] [ PDF 3645KB] ( 1795 )

	243	*Gene cloning, expression and polyclonal antibody preparation of osmotically inducible gene osmY of Salmonella enterica* serovar Typh**
		WENG Xiao-Qin, Zhang-Hong, Zhan-Li-Fang, Zhang-Xiao-Lei, Sheng-Xiu-Mei, Xu-Shun-Gao, Huang-Xin-Xiang
		Objective: To clone and express osmotically inducible gene osmY of xserovar Typhi(S. Typhi), and prepare the anti-OsmY polyclonal antibody. Methods: The DNA fragment of osmotically inducible gene osmY from S.Typhi GIFU 10007 was cloned using PCR. The sequence encoding the protein OsmY was cloned into the pET22b(+) vector and expressed in E. coli JM109. The fusion protein OsmY-His6 was purified by Ni affinity chromatography and was used as antigen to prepare polyclonal antibody. Results: The protein expression strain of osmotically inducible gene osmY was prepared successfully and was highly expressed in E. coli JM109. The antiOsmY polyclonal antibody was prepared successfully. Conclusion: This study was contributed to research on the role of OsmY in hyperosmotic stress of S.Typhi.
		2013 Vol. 23 (3): 243- [Abstract] ( 1182 ) [HTML 1KB] [ PDF 3052KB] ( 1395 )

	247	Synthesis and release property of porphyrin-cored star-shaped poly(ε-caprolactone)-block-poly(ethylene glycol)
		HUANG Ya-Fei-1, GE Yan-Ru-1, QI Xue-Yong-1, DAI Xiao-Hui-2
		Objective: In order to overcome the quenching of the traditional micromolecular porphyrin drug,the targeting polymer nanoparticles with porphyrin were obtained. Methods:Porphyrin-cored star-shaped poly(caprolactone)-block-poly(ethylene glycol) copolymers (SPPCL-b-PEO) were synthesized via ringopening polymerization and esterification reaction,and were characterized by gel permeation chromatograph,nuclear magnetic resonance spectrum,fourier transform infrared spectroscopy and ultraviolet visible spectroscopy.The photosensitizing efficiency of SPPCL-b-PEO investigated by fluorescence probe method.The selfassembly behavior of SPPCL-b-PEO was investigated by transmission electron microscope.The drug loading and release properties of SPPCL-b-PEO were studied by ultraviolet visible spectroscopy. Results:The polymeric poly(caprolactone)-block-poly(ethylene glycol)(PCL-PEO) skeleton can efficiently prevent selfquenching of the central porphyrin in the copolymers.The selfassembled aggregates changed from spherical micelles to wormlike micelles with decreasing the weight fraction of hydrophilic poly(ethylene glycol)(PEO) block in the copolymers.The Doxorubicinloaded SPPCLbPEO nanospheres possess the high drug content and entrapment efficiency, and exhibit property of pH-induced drug release. Conclusion: SPPCLbPEO will not only provide potential porphyrin core for photodynamic therapy but also improve targeting to pH5～6 tumor tissues for drug delivery.
		2013 Vol. 23 (3): 247- [Abstract] ( 1218 ) [HTML 1KB] [ PDF 5360KB] ( 2093 )

	254	Preparation and characterization of mPEG modified magnetic longcirculating epirubicin liposomes
		FANG Yu-1, CHEN Jin-2, WU Xun-Shen-1, WU Xiu-Gen-2, JIANG De-Li-1, CHEN Min-1, XIE Ji-Min-1
		Objective: To prepare new dosage form of targeted anti-cancer drug， epirubicin long-circulating magnetic liposomes. Methods: Fe₃O₄magnetic particles were synthesized by the chemical co-precipitation method. Long-circulating epirubicin liposomes were prepared by the ammonium sulphate gradients with ethanol injection, and the methoxy polyethylene glycol(mPEG) was added to modify the membrane property of the liposomes. The characteristics of liposomes were measured by transmission electron microscope (TEM), external magnetic field and FT-IR. The entrapment efficiency of epirubicin was measured by Sephadex-UV method, and the influence of preparation conditions on the encapsulation efficiency of epirubicin was studied to obtain the optimum conditions. Results: The magnetic Fe3O4 nanoparticles were nearly spherical in shape with an average diameter of around 20 nm. The longcirculating epirubicin liposomes were nearly round with an average particle of 50 nm. The entrapment efficiency of epirubicin was determined to be 575％. Meanwhile, the experiment results indicated that the long-circulating epirubicin liposomes show good magnetic response, in vitro stability and sustained release property. Conclusion: The method for the preparation of longcirculating epirubicin liposomes in this work was easy to be performed with the merits of high encapulation efficiency and reproducibility.
		2013 Vol. 23 (3): 254- [Abstract] ( 1359 ) [HTML 1KB] [ PDF 4854KB] ( 1739 )

	259	Hazards analysis of HBV infection in children of migrant workers in suburban area of Shanghai
		ZHU Yi-Bing-1, Yin-Jian-Hua-2, JI Wei-1, Liu-Hua-1, Li-Guang-Ping-1, ZHOU Yan-Ping-1
		Objective: To analysis the risk factors which associated with the hepatitis B virus transmission of migrant children in Baoshan district, Shanghai, in order to provide a scientific basis for the prevention and control of further development of the children of migrant workers hepatitis B virus (HBV) infection. Methods: The uniform prepared questionnaire was used in epidemiological survey of hepatitis B virus infection of migrant children in Baoshan district, and univariate and multivariate analysis of risk factors for hepatitis B virus infection was done in the survey results. Results: The overall infection rate was 30％ for the children of migrant workers in Baoshan District, and in both of univariate analysis and logistic regression analysis, the three consistent risk factors were mothertochild transmission, family history of hepatitis B and hepatitis B vaccination. Conclusion: The key to prevent the spread of hepatitis B virus is to strengthen the Hepatitis Brelated knowledge of migrant workers and their children′s health education, to take ahead of the immune prevention, and to cultivate health habits.
		2013 Vol. 23 (3): 259- [Abstract] ( 1107 ) [HTML 1KB] [ PDF 2682KB] ( 1399 )

	263	Clinical efficacy of tacrolimus combined with small does of hormone therapy on IgA nephropathy
		ZHANG Jun-Xian, Qian-Jian-Zhong
		Objective: To study the clinical efficacy and safety of tacrolimus combined with glucocorticoid on IgA nephropathy patients with moderate proteinuria. Methods: Twentyfive patients with IgA nephropathy were enrolled in the study and were randomly divided into two groups：single hormone group, 14 cases were given single glucocorticoid treatment; combined treatment group,11 cases were given tacrolimus combined with glucocorticoid treatment. The changes of 24 h urine protein, aminotransferase, serum creatinine, serum glucose, blood pressure, treatment effects, and side effects were observed and compared between two groups before and 10 d, 1 month, 2 month, 4 month, 6 month after treatment. Results: There were no significant differences between two groups with regard to side effects of treatment; and the effect of tacrolimus combined with glucocorticoid were better than taking glucocorticoid singly on IgA nephropathy. Conclusion: When tacrolimus blood concentration fluctuates at 3.76-4.35 μg/L, it takes effect more quickly on IgA nephropathy patients with moderate proteinuria and has higher clinical remission rate.
		2013 Vol. 23 (3): 263- [Abstract] ( 1210 ) [HTML 1KB] [ PDF 3542KB] ( 1519 )

	268
		SHANG Ru-Xia-1, 2 , SHAO Ji-2, CHEN Li-3

		2013 Vol. 23 (3): 268- [Abstract] ( 1213 ) [HTML 1KB] [ PDF 2367KB] ( 1606 )

	270
		DA Jing-Jing, QIAN Min-Zhang

		2013 Vol. 23 (3): 270- [Abstract] ( 856 ) [HTML 1KB] [ PDF 3155KB] ( 1592 )

	274
		LI Jiang-1, She-Peng- 2, Qian-Liang-Yu-2, Chao-Hua-Jun-1, Kong-Fan-Zhi-2

		2013 Vol. 23 (3): 274- [Abstract] ( 1172 ) [HTML 1KB] [ PDF 2126KB] ( 1358 )

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