The Influence of Nano-apatite on c-myc and p53 Gene in the Hepatocellular Carcinoma

The influence mechanism of the nano-apatite on the human hepatocellular carcinoma in vitro was investigated. Using the homogeneous precipitation method, the nano-apatite was synthesized at room temperature, and it was characterized with transmission electron microscopy (TEM) and the Zataplus. The influence on the expression of the c-myc and p53 gene in the human hepatocellalar carcinoma cell lines were tested with the TEM and hybridization in situ. The TEM and the Zataplus analyses show that the nano-apatite is distributed homogenously in size and needle-shaped sizes, which ranges from 67.5 nm to 88.3 nm. It is found that the nanoapatitet increases the volume of the human hepatocellalar carcinoma cells, makes extensive cytoplasmic vacuolization, the mitochondria swelling, chromatin in nucleus dispersed partially and condensed around the nuclear membranes. The interspace in nuclear membranes were separated and even the cytoplasm dissolved. It is also found that the expression of the c-myc gene is inhibited, but the p53 /s enhanced. The experimental results demonstrate that the nano-apatite enables the oneosis of the human hepatocellular carcinoma cells by down-regulation of the expression of the c-myc and up-regulation of the expression of the p53 in vitro.     

纳米磷灰石对肝癌癌基因表达的影响

研究纳米磷灰石体外对人肝癌细胞系Bel-7402的作用机理。应用均相共沉淀法室温下合成纳米磷灰石，用透射电镜和电位粒度仪进行表征；利用透射电镜、原位杂交细胞化学检测其对人肝癌细胞系Bel-7402 c-myc和p53基因表达的影响。纳米磷灰石呈均匀分散的针状颗粒．粒径范围在67．5～88．3nm之间。可使肝癌细胞体积增大，胞质空泡化，线粒体肿胀；部分细胞核内染色质分散，凝集在核膜周围，呈团块状，核膜间隙不等，胞浆溶解。抑制c-myc和增强p53基因的表达。纳米磷灰石在体外通过下调c-myc和上调p53基因的表达，致肝癌细胞胀亡。     

纳米羟基磷灰石对肝癌细胞端粒酶基因表达的影响

研究羟基磷灰石（HAP）纳米粒子对Bel-7402人肝癌细胞端粒酶基因表达的影响。采用均相沉淀法制备出稳定单分散的HAP纳米粒子,应用透射电镜、电位粒度仪对其进行表征。HAP纳米粒子作用Bel-7402肝癌细胞4 h后,采用原位杂交技术检测Bel-7402肝癌细胞的端粒酶基因表达。结果表明HAP纳米粒子作用组的Bel-7402肝癌细胞的端粒酶阳性细胞比例为61.38%,而对照组的端粒酶阳性细胞比例为87.89%,2组有显著性差异（P〈0.01）。HAP纳米粒子可使Bel-7402肝癌细胞的端粒酶基因表达下调。     

羟基磷灰石纳米粒子对肝癌细胞PCNA表达的影响

为了研究羟基磷灰石(HAP)纳米粒子对肝癌细胞增殖细胞核抗原表达的影响,采用均匀沉淀法制备了均匀分散的纳米尺度的HAP纳米粒子。以0.56 mmol/L的HAP纳米粒子与Bel-7402肝癌细胞作用后,采用流式细胞技术检测细胞周期时相的变化,行免疫细胞化学法PCNA染色,形态学观察和定量分析细胞的PCNA表达。结果显示HAP纳米粒子能使Bel-7402细胞的细胞增殖周期阻滞于G1期,PCNA表达降低,与对照组比较有显著性差异(P<0.01)。HAP纳米粒子可能通过抑制PCNA的表达,起到抑制肝癌细胞增殖的作用。     

Microbiological synthesis of nanophase PbS by Desulfotomaculum sp.

The uniform nanoparticles PbS with diameters about 13 nm could be synthesized by Desulfotomaculum sp. under mild condition. The effects of the processing variables such as pH and temperature were investigated. The obtained products were in detailed by means of transmission electron microscopy (TEM) and X-ray diffraction (XRD), respectively. The results revealed that the PbS crystallites were identical in structure, shape and size under different temperatures while their morphology changed from rod to spheroidal with pH increasing. In the biological synthetic process for PbS nanoparticles, Desulfotomaculum sp. can use sulfate as a terminal electron acceptor to produce sulfide which acts as the source of sulfur for the formation of PbS nanoparticles. Supported by the Ministry of Science and Technology of the People’s Republic of China, and the Key Technologies R&D Program of China (Grant No. 2001BA540C)     

Cytotoxinic Mechanism of Hydroxyapatite Nanoparticles on Human Hepatoma Cell Lines

1　IntroductionWiththedevelopmentofthenanometerscienceandtechnology ,theuseandstudyofthenano biomaterialsonmedicalsciencebegintoshowitsbrillianttalents .Itwasfoundthatsomeinorganicnanoparticles[1] havethenano biologicaleffectascomparedwithnon nanometer[2 ] .Whenthoseinorganicnanoparticlesaresmalltothenano level,theycaninhibittheproliferationofcancercells ,andatthesametimetheyaffectedthenormalcellslittle .Tofur therconfirmthebiologicalcharacterofthenanoparticle ,thefollowingexperimentshavebeenp…     

p53基因在小鼠睾丸发育过程中的表达规律

以17组出生后不同发育阶段的昆明种正常小鼠睾丸组织为材料,利用地高辛标记的p53基因探针在其石蜡组织切片上进行DNA-mRNA分子原位杂交,探讨p53基因在小鼠睾丸生后发育全过程中的表达规律.结果发现：p53基因的首次表达出现在生后第15 d的初级精母细胞中,在睾丸后续发育至57 d的全过程中,初级精母细胞中均有表达信号;次级精母细胞和圆形精子细胞至生后27 d时也开始出现表达;而精原细胞和精子中始终均无阳性表达信号.p53基因的表达活跃期分别出现在生后20 d、33 d和50 d.上述结果表明：p53基因在小鼠睾丸生后发育过程中发挥着重要的作用;其表达活跃期和凋亡高峰期基本同步.     

乳腺浸润性导管癌p53表达与腋窝淋巴结转移的相关性分析

为研究乳腺浸润性导管癌组织p53表达及腋窝淋巴结转移的相关性,作者利用免疫组化染色检测72例浸润性导管癌标本中p53的表达水平,分析p53表达及腋窝淋巴结转移的关系。得出结果:腋淋巴结转移组共37例(52%),p53阳性表达21例(57%);未转移组共35例(48%),p53阳性表达11例(31%),转移组p53阳性表达较未转移组明显增高(P0.05);淋巴结转移数1~3个25例(68%)、≥4个12例(32%),p53表达阳性分别是12例(48%)和8例(67%),p53表达阳性率与淋巴结转移数呈正相关(P0.05)。结果表明,通过检测乳腺浸润性导管癌组织中p53蛋白的表达水平可以判断腋窝淋巴结转移状况和愈后。     

肝癌抑制基因-1转运功能区的研究

肝癌抑制基因-1(hepatocellular carcinorna suppressor 1,HCCS1)是一种潜在的肝癌抑制基因,在细胞内发挥蛋白分拣运输的作用,其抑癌作用有可能与蛋白转运的功能有一定的关系.本研究旨在寻找HCCS1序列中与转运功能相关的序列区域.首先通过亚克隆技术,构建以pEGFP-C2为载体,对不同长度HCCS1-cDNA片段进行亚克隆,将构建的亚克隆转染到HeLa细胞,通过免疫荧光显微镜观察不同长度HCCS1蛋白在细胞内的分布,及其与6-磷酸甘露糖受体(M6PR)的共定位.经过PCR及亚克隆后,目的片段成功插入载体质粒,构建了以pEGFPC2为载体的含有4个不同长度HCCS1片段的亚克隆.分别是:pEGFP-C2-N1836,pEGFP-C2-N1572,pEGFP-C2-1707和pEGFP-C2-1506,其中pEGFP-C2-N1836和pEGFP-C2-N1572编码的HCCS1蛋白,长度不同均呈颗粒状、极性分布于核周的胞质内,且与M6PR有共定位;而pEGFP-C2-1707片段编码的HCCS1蛋白虽然也呈颗粒状分布于核周,但极性分布消失,且与M6PR共定位消失;pEGFP-C2-1506与pEGFP-C2-1707结果相似.结果显示了HCCS1-cDNA的5'端的129 bp序列为与HCCS1的极性及M6PR共定位相关的区域.     

Titanium dioxide nanoparticle absorbed by hepatoma cells in vitro

It is reported that nanoparticles can be applied as carriers and anti-cancer medicines. But the interaction of nanoparticles and cells is unclear. The purpose of this study was to discuss whether inorganic crystal nanoparticles can get through cells with intact crystal. BEL7402 hepatoma cells and titanium dioxide (TiO₂) nanoparticles were selected and incubated together in vitro. All specimens were prepared and observed under a transmission electron microscope (TEM). TiO₂ nanoparticles were found not in the nuclear area but in the cytoplasma. TiO₂ nanoparticles maintained the plate-like shape during absorbing. The result shows that hepatoma cells can endocytose the intact TiO₂ crystal nanoparticles. It implies that novel nano-effect plays an important role in the biomedicinal application of inorganic crystal nanoparticles. Funded by National Natural Science Foundation of China (No. 50472040)     

Inhibition of the growth of human hepatoma cell line both in vitro and in vivo by transducing CKI gene p21WAF-1 with GE7 targeting gene delivery system

The EGF receptor-mediated targeting gene delivery system GE7 was used to transduce exogenous gene pCEP-p21WAF-1 into human hepatocellular carcinoma cell both in vitro and in vivo. After in vitro transduction of the exogenous gene, the growth of the cell lines SMMC-7721 and BEL-7402 was significantly inhibited compared with the control. On day 8 the inhibition rates of the above cell lines reached 56.0% and 66.7%, respectively. The in vivo experiment showed that the growth of human hepatoma transplanted in nude mice injected with GE7 gene delivery system subcutaneously once a week for 3 weeks was remarkably inhibited compared with that of untransfected control. The average tumor weight of the experiment group was (0.083 ± 0.043) g, while that of the control group was (0.281± 0.173) g. The difference is significant (P<0.05). It was indicated that GE7 gene delivery system could efficiently transduce exogenous gene pCEP-p21WAF-1 into hepatoma cell with high EGF receptor expression, and inhibit the cell growth with high efficacy both in vivo and in vitro.     

Preparation and characterization of cationic PLGA-PEGLf/DOPE nanoparticles for HO-1 gene delivery

Cationic nanoparticles(NPs)for gene delivery were successfully prepared by assembling carboxylation poly(lactic-co-glycolic acid)(PLGA),polyethylene glycol(PEG),L-α-Phosphatidylethanolamine(DOPE)and octadecyl quaternized carboxymethyl chitosans(OQCMC).Lactoferrin(Lf)was selected as a targeting ligand conjugated to PLGA via bifunctional PEG,yielding PLGA-PEG-Lf/DOPE NPs to be used for gene vectors.Fourier transform infrared spectroscopy(FTIR),UV and nuclear magnetic resonance(NMR)spectroscopy were performed to evaluate the synthesis of the vectors.The characteristics of the vectors loaded heme oxygenase(HO-1)gene were evaluated by transmission electron microscope(TEM),particle size analyser and fluorescent microscopy.The experimental results showed that the obtained vectors were spherical in shape with average particle size of 142.2 nm and zeta potentials of +16.4 mV.The vectors could protect the loaded gene from the degradation by nuclease.For 293T cells,there is high transfection eff iciency of the vectors similar to liposome-2000.It can be concluded that the established cationic PLGA-PEG-Lf/DOPE NPs have potential gene delivery ability for gene therapy.     

Synthesis and properties of water-soluble CdSe/ZnS nanocrystals with the type-I core/shell structure

The synthesis of CdSe/ZnS core/shell nanocrystals though aqueous phase using the coprecipitation method was reported. The influences of factors such as injection methods and dosages of precursors, reaction duration of water-bathing and the initial CdSe：ZnS molar ratio were discussed. In comparison to the CdSe plain core nanocrystals, the CdSe/ZnS core/shell nanocrystals show much brighter photoluminescence demonstrated by the photoluminescence spectra. The epitaxial growth of the core/shell structures was verified by TEM and XRD.     

TGF—β1对肝癌细胞系Hep3B中干性相关基因表达的影响

有研究表明TGF-β1可以诱导诸多上皮来源的癌细胞和正常细胞发生EMT并使其功能发生改变。实验就TGF-β1对肝癌细胞系Hep3B的作用展开研究：通过CCK8检测TGF-β1对Hep3B细胞增殖的影响,RT-PCR实验检测TGF-β1处理后细胞中EMT及干性相关基因的表达变化。结果表明：TGF-β1对Hep3B细胞的增殖无抑制作用;TGF-β1处理Hep3B细胞6d后EMT相关基因的mRNA表达水平并无显著改变,但TGF-β1可上调Hep3B细胞干性基因Oct-4,Klf-4,Nanog,C-myc的表达,并下调分化基因albumin的表达。结果提示TGF-β1一定程度上影响肝癌细胞系的干性基因表达,但并不一定是以发生EMT为前提的。     

Study on the Variation of Calcium and Phosphorus in Nano Hydroxyapatie Treated Cancer Cells through Synchrotron Radiation XRF

1Introduction Synchrotronradiation(SR)isanexcellentsource withhighbrightnessforX rayfluorescence(XRF).Syn chrotronradiationX rayisappliedaslightresourceofSR XRF.TheSR XRFisgeneratedbyhighenergyelectron accelerator.Itoffershighsensitivityandspatialresoluti…     

Characterization of tricalcium phosphate solubilization by Stenotrophomonas maltophilia YC isolated from phosphate mines

The phosphate solubilizing characteristics of a strain YC, which was isolated from phosphate mines (Hubei, China), were studied in National Botanical Research Institute’s phosphate (NBRIP) growth medium containing tricalcium phosphate (TCP) as sole phosphorus (P) source. The strain YC is identified as Stenotrophomonas maltophilia (S. maltophilia) based upon the results of morphologic, physiological and biochemical characteristics and 16S rRNA sequences analysis. The results show that the strain S. maltophilia YC can solubilize TCP and release soluble P in NBRIP growth medium. A positive correlation between concentration of soluble P and population of the isolate and a negative correlation between concentration of soluble P and pH in the culture medium are observed from statistical analysis results. Moreover, gluconic acid is detected in the culture medium by HPLC analysis. It indicates that the isolate can release gluconic acid during the solubilizing experiment, which causes acidification of the culture medium and then TCP solubilization. S. maltophilia YC has a maximal TCP solubilizing capability when using maltose as carbon source and ammonium nitrate as nitrogen source, respectively, in NBRIP growth medium. Foundation item: Project(2004CB619201) supported by the Major State Basic Research and Development Program of China; Project(Z200515002) supported by the Key Project Foundation of the Education Department of Hubei Province, China; Project(GCP200801) supported by the Open Research Fund of Key Laboratory for Green Chemical Process of Ministry of Education, China; Project(Q200811) supported by the Youths Science Foundation of Wuhan Institute of Technology, China     

模拟体液中纳米羟基磷灰石/壳聚糖的制备及表征

从仿生合成的思路出发,分别以模拟体液和含有壳聚糖的模拟体液为反应介质,通过磷酸和硝酸钙反应合成了羟基磷灰石(HAp)和HAp/壳聚糖复合粉体,利用TG-DTA、XRD、FT-IR和TEM等对HAp和HAp/壳聚糖的形成过程、结构及其微观形貌进行了研究.结果表明,在模拟体液中合成的HAp粉体呈现出球状和短棒状形态;HAp/壳聚糖复合粉体呈现出不规则形状,粒度<100 nm,主要晶相为羟基磷灰石.体外生物活性实验结果表明,HAp/壳聚糖复合材料比纯HAp具有更好的生物活性,具有较强的诱导磷灰石沉积能力.     

负载型纳米ZrO2/Al2O3复合载体的制备与表征

以大孔Al2O3为基载体，用沉积-沉淀法和溶胶-凝胶法分别制备负载型纳米ZrO2／Al2O3复合载体，利用X射线衍射(XRD)、透射电子显微镜(TEM)、比表面与孔径测定仪等对载体和催化剂进行了表征．结果表明，负载型纳米ZrO2／Al2O3复合载体具有较大的比表面积和适宜的孔径分布，ZrO2第二载体在基载体上分布较均匀，粒径分别为19和15nm，复合载体的比表面积分别为101．6和113．5m^2／g．