OBJECTIVE:To prepare a novel folate-targeted magnetic nanocomposites loaded with tissue facor pathway inhibitor 2 (TFPI-2) and cisplatin (CDDP) and to investigate its targeting ability and anti-tumor effect on nasopharyngeal carcinoma HNE-1 cells in vitro.METHODS:The copolymer folic acid-polyethylene glycol-polyethyleneimine (FA-PEG-PEI) was synthesized through amidation reaction, and then FA-PEG-PEI/ magnetic nanoparticles-CDDP/TFPI-2 (MNP-CDDP/TFPI-2) nanocomposites was obtained by electrostatic adsorption between TFPI-2 plasmid and magnetic nanoparticles loaded with CDDP (MNP-CDDP) with vortex FA-PEG-PEI. (1)H Nuclear Magnetic Resonance ((1)H NMR ) was used to determine if FA-PEG-PEI was synthesized. The particle size, zeta potential and morphology were detected by dynamic light scattering (DLS) and transmission electron microscope (TEM). The content of Fe and CDDP was measured by phenanthroline and o-phenylenediamine (OPDA) colourimetry. Agarose gel electrophoresis was used to analyze the binding ability of FA-PEG-PEI/MNP-CDDP to TFPI-2 plasmid. Molecular targeted uptake of FA-PEG-PEI/ MNP-CDDP/TFPI-2 coupling with green fluorescent protein (GFP) in NPC cells were observed by Prussian-blue iron staining and fluorescence microscope. The levels of TFPI-2 protein expression after transfection were evaluated by Western blot. The effects of nanocomposites on HNE-1 cells proliferation and apoptosis were measured with Cell Counting Kit-8(CCK-8) and flow cytometry.RESULTS:Special peak value of FA, PEG and PEI were showed on (1)H NMR spectrogram. The mean size and zeta potential of FA-PEG-PEI/MNP-CDDP/TFPI-2 were 141.1 nm and 21.5 mV. The nanocomposites showed a good monodispersity and an insufficient size uniformity under TEM. The content of Fe and CDDP were 116.2 μg/ml and 92.88 μg/ml, respectively. Agarose gel electrophoresis showed TFPI-2 could be encapsulated completely and protected from digestion of DNA enzyme as the mass ratio of FA-PEG-PEI/ MNP-CDDP and TFPI-2 plasmid was equal or higher than 1∶1. More blue-stained magnetic granulars and green fluorescence were seen in folate receptor (FR)-positive HNE-1 cells than in FR-negative CNE-2 (P<0.05) under microscope and fluorescence microscope. The level of TFPI-2 protein expression in HNE-1cells increased significantly after transfection by FA-PEG-PEI/ MNP-CDDP/TFPI-2, compared with other control groups (FA-PEG-PEI/MNP-CDDP group and TFPI-2 group), all P<0.05. The nanocomposites inhibitory effect on HNE-1 including cell growth inhibition rate (64.00%) and apoptosis rate (49.61%) were significantly higher than that in FA-PEG-PEI/MNP group (8.19%, 9.26%), FA-PEG-PEI/TFPI-2 group (40.35%, 19.85%) and FA-PEG-PEI/MNP-CDDP group(56.15%, 36.46%)(P<0.05).CONCLUSION:FA-PEG-PEI/MNP-CDDP/TFPI-2 nanocomposites was successfully synthesized using amidation and electrostatic adsorption technology and has a good molecular targeting and inhibitory effect on FR-positive HNE-1cells in vitro.

目的 探讨核转录因子(E2F-1)和细胞周期因子(CyclinD1)在子宫内膜癌发生、发展中的作用.方法 采用免疫组织化学SP法检测60例子宫内膜癌、32例正常子宫内膜组织中E2F-1、CyclinD1的表达.结果 正常子宫内膜标本中E2F-1、CyclinD1均无表达,子宫内膜癌组织中E2F-1与CyclinD1的阳性表达率分别为58.33％ (35/60)、60.00％ (36/60),子宫内膜癌组织中E2F-1、CyclinD1的表达均高于正常子宫内膜组织(P均＜0.05).两者的表达与肌层浸润程度、淋巴结转移、临床分期及病理分期有关(P均＜0.05).E2F-1与CyclinD1的表达呈正相关(P＜0.05).结论 子宫内膜癌组织中E2F-1、CyclinD1均呈高表达,且与子宫内膜癌的发生、发展和转移密切相关.

Objective To determine the expression of E2F-1 and CyclinD1 in the endometrial carcinoma.Methods 32 cases of normal endometrial tissue were detected by immunohistochemistry S-P.Results The expressions of E2F-1 and CyclinD1 were located in the cytoplasm and the endometrial.The positive expression rates were 58.33%(35/60) and 60.00%(36/60) which were significantly higher than that in normal endometrial tissue in 0,0(P0.05).All of their expressions were related with myometrial invasion,lymph nodemetastasis,clinical stage and pathological staging(P0.05).In endometrial carcinoma,the expressions of E2F-1 and CyclinD1 were positively correlated with each other(P0.05).Conclusion E2F-1 and CyclinD1 were highly expressed in the endometrial cancinoma and their expressions were related with endometrial carcinoma the development and the metastasis.

Objective To observe the histomorphology effects and biomechanical abilities of Chinese herbal medicine invigorating the kidney(Guling Pian) on osteoporotic(OP) ovariectomized(OVX) rats model,and explore its therapeutic mechanism.Methods 36 femal 3-month-old SD rats were randomized into sham-operated,OVX,Guling Pian low,medium,high dosage and ethynylestradiol groups,6 in each group.The dynamic and static parameters of lumbar vertebrae were tested by bone morphometry,the maximum load and rigidity of lumbar vertebrae were determined by biomechanical indentation testing 4 months later.Results ①The bone trabecula resolution,bone formation rate(BFR)/BS and BFR/BV,the number of osteoclast per mm in different dosages of Guling Pian and ethynylestradiol groups were significantly decreased than those in OVX group(P<0.01).The BFR/TV in moderate dosage Guling Pian group and bone mineral apposition rate(MAR) in middle and high dosages of Guling Pian groups were significantly decreased(P<0.01).The bone trabecula area percentage,thickness and number in moderate and high dosages of Guling Pian and ethynylestradiol groups obviously increased,the bone trabecula thickness of moderate dosage of Guling Pian group was higher than that in ethynylestradiol group(P<0.05).②The rigidity of ethynylestradiol and different dosages of Guling Pian groups and the maximum load in moderate and high dosages of Guling Pian groups were significantly higher than those in OVX group(P<0.01).The maximum load of high dosage of Guling Pian group was significantly different from that of ethynylestradiol group(P<0.05).Conclusions Guling Pian has double effects on restraining bone absorption and promoting bone formation,may decrease bone turnover to improve bone quality and to increase bone biomechanics intensity.

Objective To observe the effect of sinomenine on the proliferation of fibroblast-like synoviocytes (FLS) and expression level of MMP-3 secreted by FLS in rheumatoid arthritis in vitro. Methods FLS were obtained by digesting synovial tissues with collagens and were divided into four groups:sinomenine high, middle, low concentration and control group. The proliferation of FLS was assessed by methyl- thiazolyl-tetrazolium (MTT) assay and the mRNA expression of MMP-3 was measured by semi-quantitative RT-PCR respectively after treated by sinomenine. Results The rate of FLS proliferation was significantly reduced in groups treated by sinomenine. Compared with control group, the mRNA expression of MMP-3 markly decreased in sinomenine groups (P <0.05) and especially in sinomenine high group (P <0.01). Conclusion Sinomenine can effectively inhibit the proliferation of FLS and reduce the mRNA expression of MMP-3 of FLS in rheumatoid arthritis, that may be a mechanism in therapy of rheumatoid arthritis by sinomenine.

Objective To investigate the effect of Chinese herbal medicine which invigorating the kidney function (Gu Ling pian) on the changes in two and three dimensional structure, and bone strength in the osteoporotic ovariectomized (OVX) rat model. Methods 18 female 3-month-old SD rats were randomized into six groups(3 for each): the shame-operating group, ovariectomized(OVX) group, Gu Ling Pian(GLP) low, medium, high dosage groups and ethynylestradiol treated group . The two-dimensional and three-dimensional parameters were measured by bone histomorphometric analysis and indentation testing, respectively. 3D structure was reconstructed from serial images. Results Compared with the OVX rats, remarkable increase of the percentage of Tb.Ar, Tb.N, Tb.Th, Volume Voxels, Volume Percentage and remarkable decrease of Tb.Sp were found in GLP medium and high dosage groups. The improvement in the GLP medium and high dosage groups was more conspicuous than that in the ethynylestradiol treated group. Conclusion Chinese herbal medicine which invigorating the kidney function has obvious effects on microstructure of bone trabecula, especially on bone density in the Gu Ling Pian high dosage group.

Objective To investigate the effect of kdney-tonifying tranditional Chinese medicine,Gu Ling Pian(GLP)on regulating human osteoblasts function and the osteoprotegerin(OPG)/receptor activator of nuclear factor kappa B ligand(RANKL)expression via p38 MAPK pathway in vitro.Methods Human osteoblast were cultured with serum obtained from rat fed with high,medium,or low dose GLP.The expression levels of the OPG and RANKL mRNA were determined by real-time RT-PCR.Total p38 and phosphorylated p38 were detected by western blot.Mineralized bone nodule was detected by alizarin red staining method.Results GLP serum promoted osteoblasts differentiation,increased the formation of mineralized bone nodule,up-regulated the OPG expression while RANKL expression decreased and increased the total protein of phosphorylated p38.Conclusion Kdney-tonifying traditional Chinese medicine,Gu Ling Pian,induced osteoblasts differentiation and synthesis of OPG/RANKL probably through p38 MAPK pathway.

霉菌性阴道炎为妇科门诊常见病、多发病.2002年6月-2004年7月,本院采用定君生治疗霉菌性阴道炎取得较好效果,现报道如下.