Specific anti-tumor effect induced by attenuated <Emphasis Type="Italic">Salmonella typhimurium</Emphasis> vaccine expressing extracellular region of vascular endothelial growth factor receptor 2

The purposes of this research were to study the stable expression of exogenous gene encoding therapeutic protein in attenuated Salmonella typhimurium, observe the metabolism of oral gene vaccine carried by attenuated Salmonella typhimurium in BALB/c mouse, and investigate the feasibility of prevention and treatment of tumors by the recombinant bacteria. Recombinant plasmid pcDNA3.1＋ VEGFR2（n1-7） was transformed into competent attenuated Salmonella typhimuriurn SL3261 to develop oral DNA vaccine SL3261-pcDNA3.1＋VEGFR2（n1-7）. To observe whether the exogenous gene can be expressed in the recombinant bacteria, PCR was performed to amplify the CMV promoter of the eukaryotic expression vector as the proof of stable expression of exogenous protein; transmission elec- tron microscopy （TEM） was applied to observe the morphology of the recombinant bacteria to confirm that the exogenous gene has no impact on the growth of the bacteria, and then BALB/c mice were immunized with the gene vaccine. After inoculation of the gene vaccine, the recombinant bacteria SL3261 could be detected in the tissues such as small intestine, colon, liver and spleen. And then, mice in each group were challenged with tumor cells. The results of animal experiment showed that tumor growth of the mice in experimental group was inhibited and survival time of immunized mice was prolonged compared with control groups. A higher lymphocyte infiltration in tumors from animals treated with DNA vaccine was observed. Immunohistochemical analysis of tumor samples revealed an enhanced accumulation of CD8^＋ cytotoxic T lymphocytes, as well as an increase in CD4^＋ cells in the tumore of animals treated with the oral gene vaccine compared to tumors from control group mice. UI- trestructure of the tumor tissue showed that tumor cells in the samples of the immunized mice were well-differentiated. Our research confirmed that the exogenous gene can be stably expressed in the attenuated Salmonella typhimurium and has no impact on the growth of the r     

CpG DNA enhances the immune responses elicited by the DNA vaccine against foot-and-mouth disease virus in guinea pigs

CpG DNA is DNA sequence that has immune stimulatory effects. Several lines of investigation over the past few years indicate that CpG DNA plays an important role in the induction of immune responses to DNA vaccines. In this study, CpG DNA-containing synthetic oligodeoxynucleotide (CpG-ODN) was cloned into the eukaryotic expression plasmid encoding a fusion protein containing b- galactosidase from E. coli and immunogenic epitopes of foot- and-mouth disease virus (FMDV) type O, and the immune responses induced by the plasmid were assayed. The results showed that guinea pigs immunized with the recombinant plasmid containing CpG-ODN generated a higher level of FMDV-neutralizing antibody and a stronger T cell proliferative response and protection against viral challenge than those receiving the plasmid containing no CpG-ODN. Our study demonstrated that it is an effective route to enhance the efficacy of DNA vaccines by inserting exogenous CpG DNA into the plasmids, and the DNA vaccine developed here is a promising candidate to prevent FMDV infection.     

重组戊型肝炎疫苗的免疫原性试验

目的　研究重组戊型肝炎疫苗的免疫原性。方法　通过对小鼠、新西兰兔的多点皮内注射及肌肉注射 ,然后测定其抗体效价 ,研究重组戊型肝炎疫苗的性能。结果　小鼠在免疫 7d后开始产生抗体 ,两周抗体阳性率为80 % ,四周抗体阳性率达 10 0 %。新西兰兔从第二周后开始产生抗体 ,第三周达到高峰 ,抗血清在 1∶6 4 0 0稀释度的阳性率达 4 0 %。结论　重组戊型肝炎疫苗免疫动物具有良好的免疫原性 ,为进一步研究开发戊肝疫苗对人体的免疫提供了基础     

Integral Points on a Class of Elliptic Curve

0 IntroductionZasegairerc[h1]fodre sbcirgibientde gsreavle rpaolin tmse tohnocdsert waihnicehlli ppetircm citur ovnese btyogiving the upper bound of solution. Unfortunately,this upperbound was verylarge andsometi mes beyondthe range of com-puter searching.For a particular elliptic curvey2=x3-30x+133(1)he mentioned he can find all integral points and the largestpoint is (x,y) =(5 143 326 ,±11 664 498 677) by using Mas-ser and W櫣stholz bounds on elliptic logarithms .Although recent results on…     

The Escherichia coli O157:H7 DNA detection on a gold nanoparticle-enhanced piezoelectric biosensor

This paper presents development of a quartz crystal microbalance （QCM） biosensor for real-time detection of E. coil O157：H7 DNA based on nanogold particles amplification. Many inner Au nanoparticles were immobilized onto the thioled surface of the Au electrode, then more specific thiolated sin- gle-stranded DNA （ssDNA） probes could be fixed through Au-SH bonding. The hybridization was induced by exposing the ssDNA probe to the complementary target DNA of E. coli O157：H7 gene eaeA, then resulted in a mass change and corresponding frequency shifts （ △f ） of the QCM. The outer avidin-coated Au nanoparticles could combine with the target DNA to increase the mass. The electrochemical techniques, cyclic voltammetry （CV） and electrochemical impedance spectroscopy （EIS） were adopted to manifest and character each step. The target DNA corresponding to 2.0×10^3 colony forming unit （CFU）/mL E. coil O157：H7 cells can be detected by this biosensor, so it is practical to develop a sensitive and effective QCM biosensor for pathogenic bacteria detection based on specific DNA analysis. The piezoelectric biosensing system has potential for further applications, such as food safety and environment monitoring, and this approach lays the groundwork for incorporating the method into an integrated system for in-field bacteria detection.     

Construction of multivalent DNA vaccines forMycobacterium tuberculosis and its immunogenicity

The coding regions of Ag85B MPT-64, and ESAT-6 secreted proteins were cloned initially into the eukaryotic expression vector pJW4303, then transformed to E. coli Top 10 strain for plasmid DNA extraction and further analysis. Plasmids containing the right insertion were sequenced to confirm their identity. COS7 cells were transfected with a mixture containing serially diluted plasmid DNA encoding three secreted proteins and Lipofectin (Gibco). The supernatants and pellets prepared from various cell lines were run on SDS-PAGE gel and the expression of these proteins in COS7 cells were demonstrated by immunoblot using polyclonal or monoclonal antiserum of M.TBH37Rv. 21 days after first vaccination of C57BL-6 mice by all three recombinant eukaryotic expressing vectors, antibody titer for Ag85B reached 1∶3200. 21 days after second vaccination, the antibody titer reached 1∶102400. The highest antibody levels induced by multivalent vaccines after the second injection were equal to or even greater than the highest antibody levels of single DNA vaccine reported in literature after third injections. Antibody titer of MPT-64 was 1∶50 after the first injection and it reached 1∶200 after the second injection. No antigen-specific antibody against ESAT-6 was detected in sera harvested from immunized mice 21 days after both injections. Antigen-specific IFN-g level of Ag85B was 110 pg/mL while no antigen-specific IFN- g level of ESAT-6 and MPT-64 was detected even after third injections. To our knowledge, it is the first time that studies of polyvalent recombinant DNA vaccines against TB were carried out in C57BL-6 mice. Our results indicated that multiple DNA vaccines could be used to enhance protective responses against M.TB.     

不同轮伐期对巨尾桉人工林碳固存的影响

【目的】深入探讨不同轮伐期对巨尾桉人工林碳固存的影响,为从应对全球气候变化的视角确定合理轮伐期提供理论依据。【方法】以轮伐期为短(7a)、中(13a)和长周期(21a)的巨尾桉人工林为研究对象,通过对不同轮伐期桉树林分生物量、碳固存、年平均固碳量的分析,揭示不同轮伐期对桉树林分碳固存的影响。【结果】巨尾桉人工林的生物量碳随着轮伐期的延长而逐渐增加,由7a轮伐期的(75.81±5.12)t·C/hm~2增至13a轮伐期的(180.11±19.97)t·C/hm~2以及21a轮伐期的(192.08±16.50)t·C/hm~2,方差分析表明,13a和21a轮伐期的总生物量碳显著高于7a轮伐期,而13a和21a轮伐期之间的差异不显著。巨尾桉人工林土壤有机碳随轮伐期延长而显著降低,由7a轮伐期的(89.99±0.35)t·C/hm~2、13a轮伐期的(85.42±0.76)t·C/hm~2下降到21a轮伐期的(74.64±0.24)t·C/hm~2。7~13a仍是巨尾桉人工林固碳能力迅速增长期,年平均总生物量碳由7a时的10.78t·C/(hm~2·a)迅速提高到13a的19.54t·C/(hm~2·a),增长81%;21a时巨尾桉人工林进入固碳能力下降期,年平均总生物量碳降至3.78t·C/(hm~2·a),固碳能力只是13a的19.34%。【结论】在南亚热带,巨尾桉人工林的最佳轮伐期确定在13a左右较为适宜,这与经济效益的最大化一致。     

Cloning,expression of<Emphasis Type="Italic">Crocus sativus</Emphasis> phytoene desaturase gene and preparation of antiserum against it

A 2 149 bp full length phytoene desaturase (PDS) cDNA was first cloned from saffron (Crocus sativus L.) stigma using RT-PCR technique and a rapid amplification of cDNA end (RACE) strategy. The cDNA has an open reading frame of 1 697 bp, which encodes a polypeptide of 565 amino acids. The coding region of the cDNA was inserted into a prokaryotic expression vector pET-21a(+) and over-expressed inE. coli BL21 (DE3). The fusion proteins were found largely in an insoluble inclusion bodies. The purified fusion protein was used to immunize rabbits to obtain polyclonal antiserum with titer of 1×10⁵. Western blot analysis by using this particular antiserum showed that the higher expression level of PDS in mature stigma than in leaves and stamen, and the higher expression level of PDS in mature stigma than in young stigma. Foundation item: Supported by the Doctoral Foundation of the Ministry of Education, P. R. China and the Young Science Foundation of Sichuan University (Grant 0020405505012) Biography: Bai Jie (1968-), female, Ph. D candidate, research direction: plant developmental biology and reproductive engineering.     

Vanadyl ions binding to GroEL （HSP60） and inducing its depolymerization

Several vanadium compounds have been known for the hypoglycemic and anticancer effects. However, the mechanisms of the pharmacological and toxicological effects were not clear. In this work, we in- vestigated the potential targets of vanadium in mitochondria. Vanadyl ions were found to bind to mi- tochondria from rat liver with a stoichiometry of 244±58 nmol/mg protein and an apparent dissocia- tion constant (Kd) of (2.0±0.8)×10·16 mol/L. Using size exclusion chromatography, a vanadium-binding protein was isolated and identified to be the 60-kDa heat shock protein (HSP60) by mass spectrometry analysis and immunoassays. Additionally, binding of vanadyl ions was found to result in depolymeri- zation of homo-oligomeric HSP60 (GroEL). HSP60 is an indispensable molecular chaperone and in- volved in many kinds of pathogenesis of inflammatory and autoimmune diseases, e.g. type 1 diabetes. Our results suggested that HSP60 could be a novel important target involved in the biological and/or toxicological effects of vanadium compounds.     

The study on the immune response induced by expressing recombinant plasmid of dengue virus type 2 NS3 protein

The PSV·NS3, an expressing recombinant plasmid of dengue virus type 2 NS3 protein, was injected directly into the quadriceps of Balb/C mice to explore whether it could inducing immune response. The splenic T cell subsets of two groups was analysed by flow cytometry. It was found that the percentage of CD4⁺ and CD8⁺ T cells of experimental group were significantly higher than those of the control group. The titer of IgG antibody was as high as 1∶5 120 in experimental group, but it couldn’t be detected in control group by ELISA. The western blot further proved that the IgG antibody was specific for NS3 protein. Those results Suggested that inoculation Balb/C mice with PSV·NS3 could inducing immune response, and the NS3 protein might be used as the candidate protein of DNA vaccine of dengue virus. Foundation item: Supported by Foundation of American Chinese Medicine Biography, LI Xiang-qun(1963−), male, M.D, Research direction: molecular and clinical virology.     

Sensitive Determination of DNA by RLS Enhancement of Metal Ions

0　IntroductionThequantitativeanalysisofnucleicacids,especiallythemi cro determinationofnucleicacids,isbecomingmoreandmoreimportantinmanybiologicalstudies.Recently ,apromisingspectraltechnique ,whichwasbasedonthemeasurementofen hancedresonancelightscattering (RLS) [1 ,2 ] ,hasgivenrisetostronginterestbyanalystsandbiochemistsfornucleicacidsandproteinassay[3 9] .Uptonow ,manykindscompoundshavebeenfoundRLSenhancementwhilebindingtoDNA ,andallthesecompoundsarecharacterizedofpositivechargewhicha…     

Effect of Quercetin on Breeding and Apoptosis of Cervical Cancer HeLa Cell and on Growth of Transplanted Tumor in Nude Mice

Effect of quercetin on HeLa cell system of cervical cancer was studied by methods of MTT and Annexin V-FITC/PI. The results show that quercetin has functions of inhibiting breeding of HeLa cells and inducing apoptosis of the cells. The total apoptosis rate is positively proportional to reaction duration and concentration of quercetin used. The maximum apoptosis rate being （88.76±2.35）% was obtained when the concentration was 50.0 μmol/L and the cells were treated with quercetin for 72 hours. Based on establishing a model of tumor of cervical cancer transplanted into nude mice, quercetin of different concentrations was injected into abdominal cavity of nude mice and situation of tumor growth was reviewed. The result showed that with quercetin concent＇ration increasing from 0 to 100.0 μmol/L, the transplantation volume and weight of the tumors decreased from （279.59±70.58） mm^3 and （0.145±0.019） g to （128.72±36.12） mm^3 and （0.089± 0.019） g respectively, while apoptosis rate of the transplanted tumor increased from （9.63±1.85）% to （34,98±0.47）%, which proved that quercetin inhibited increment of volume and weight of transplanted tumor in nude mice bodies.     

Fluorescence Determination of Artemisinin Using Hemoglobin as Catalyst and Pyronine B as Substrate

0 IntroductionMaalnadria siusb atr ompajicosr .he aAltrthe pmrisoibnlienmi (n qtihneghtraoopsiucs,QHS,Fig.1) is a sesquiterpene endoperoxide isola-ted fromArtemisia annuaL., an ancient Chineseherbal medicine usedfor treatment of fever and ma-laria.Studies of the structure and activity relation-ship have shownthat endoperoxide groupis essentialfor anti malarial activity of QHS and absence of thismoiety lead to completely loss in activity of thedrug. Many techniques have been developed to de…     

Synergistic interaction and gelation in cationic guar gum-sodium alginate system

The synergistic interaction between the cationic guar gum (the ammonium hydroxy-propyl-trimethyl chloride of guar gum) and sodium algirmte has been studied. The effects of the mass ratio of them, mixed temperature, balk salt ion concentration, incubation time and pH value on gelation were investigated. It has been observed that there was a gel strength maximum when the mass ratio was 0. 6, the mixed temperature was 70 ~C, the balk salt ion concentration was 1.0 mol ~ L-1 , the incubation time was 30 rain and the pH value was 8. Interaction between molecules of these two polysaccharides was investigated by FT-IR spectrometry.     

Myoglobin as Mimetic Enzyme and Its Analytical Application in Determination of H2O2

0 IntroductionSltougdiiceasl csoynstceermnsin gharveeac btieveen o txhyeg ei nm psopretcaienst ( oRbjOecSt) iinn bbiioo--medicine and analytical chemistry in recent years[1-4]. Manyi mportant life phenomena and diseases are relatedtotheinter-mediates of ROS[5-7].ROS have beeni mplicated as ani mpor-tant causative factor in cell damage,including apoptosis andnecrosis . Their proposed actions comprise lipid peroxidation,DNAdamage,the mitochondrial respiratory chain destructionand protein mo…     

Expression of TGF-β2 in LECs of Age-Related Nuclear, Cortex Cataract and the Relationship among TGF-β2, Proliferation,Apoptosis and Transdifferentiation

0 IntroductionAsgue-br-eclaaptseudl acrat acraatcatr ianctc l,ud aems ocnogrte xwh,incuhcl,e acro ratnexdcataract and nuclear cataract are more i mportant .Cortex cataract is different from nuclear cataract inoccurring,developing and clinic display,evenin his-tology. The pathogenesis of age-related cataract hasnot been perfectly described.But it has been verifiedthat thereis alayer oflens epithelial cells (LECs) un-der vertebrates’lens anterior capsule, which is i m-portant in maintaining t…     

A tumor suppressor genefhit prokaryotic expression and identification

The recombinant expression vector pGEMD-fhit which contains full encoding region offhit gene was constructed. The recombinant was introduced into the BL21 (DE3) strain ofE. coli and induced by 1 mmol/L IPTG to express a 29×10³ polypeptide offhit fusion protein. And the 29×10³ protein was sensitive and specific in reaction with anti-fhit antibody in Western blot. Foundation item: Supported by the National Natural Science Foundation of China (39770373) Biography: SUN Yan (1975−), female, Master of science, Research direction: gene engineering     

Hausdorff Measure of Linear Cantor Set

We study the Hausdorff measure of linear Cantor set E,on the unit interval,under the strong seperated condition.We give a necessary and sufficient condition for (E) =|E|^α by using the contracting ratio and the first gap.This condition is easy to use.     

Application of an Optical Biosensor to Study the Interaction between Porphyrins and Wheat Germ Agglutinin

0Introduction Porphyrinderivativeshavebeenusedasphotosensitizersinphotodynamictherapy(PDT),anewapproachdevel opedforthetreatmentofcancer[1].Cationwater solublepor phyrinsareofconsiderableinterestowingtotheirpossiblebio medicalapplications[2].Inwhich,theinhibiteffectonthe growthmetabolismofE.coliandcancercellHL60ofmeso tetra(4N ethylacetatepyridyl)porphyrin(H2NEAE pp)and itsZn(Ⅱ)complexform(ZnNEAE pp)havebeenrecentlyin vestigated[3,4].Toenhancetumor locationandanticancerac tivitiesofdr…     

Carboxymethytl Pachymaram Up-Regulates Dendritic Cell＇s Function in Hepatitis B Virus Transgenic Mice

The effect of carboxymethytl pachymaram (CMP) on the function of dendritic cells(DCs) derived from spleens of hepatitis B virus transgenic mice are studied in vitro. The phenotypes of DCs are tested by flow cytometry (FCM), cytokines measured by ELISA. The expression of DCs’ phenotypes in HBV transgenic mice are low (CD80⁺CD11c⁺:59.12±11.53 vs 9.60±4.53, p<0.01; CD80⁺ MHC-II⁺: 44.86±12.31 vs 9.80±5.72, p<0.01, normal mice vs HBV transgenic mice), the ability of DCs stimulating T lymphocytes proliferation decreases (0.37±0.11 vs 0.20±0.11, p<0.05, normal mice vs HBV transgenic mice), levels of IL-12 and IFN-γ decrease whereas the level of IL-10 increases; CMP can enhance DCs’ ability of stimulating T lymphocytes proliferation, facilitate the secretion of IL-12 and IFN-γ, inhibit the secretion of IL-10, thus up regulates DCs function. The results show a good prospective use of CMP on the treatment of chronic hepatitis B. Biography: HOU Anji (1963–), male, Ph.D. candidate, Associate professor, research direction: clinical virology.