A drainage-enhancing device for foam fractionation of proteins

The foam fractionation of nisin from its fermentation broth was studied.Two types of devices consisting of a rubber piston and a foam riser were developed to enhance foam drainage.The separation performance of these two devices was investigated.Experimental results indicated that the second device could significantly reduce the liquid fraction of the foam leaving the column,εout,leading to a higher enrichment of the out-flow stream.As its mounting height increased from 0 to 15 cm,εout declined from 7.07‰ to...     

Simultaneous detection of seven phenolic acids in Danshen injection using HPLC with ultraviolet detector

A high-performance liquid chromatographic (HPLC) method with ultraviolet (UV) detector had been developed for simultaneous quantification of danshensu, protocatechuic aldehyde, caffeic acid, salvianolic acid D, rosmarinic acid, salvianolic acid B and salvianolic acid A in Danshen injection. According to the UV spectra of these components, three detection wavelengths have been selected as follows: 280 nm for danshensu and protocatechuic aldehyde, 326 nm for caffeic acid, salvianolic acid D and rosmarinic acid, 286 nm for salvianolic acid B and salvianolic acid A. The limit of detection (LOD) was improved to be in the range of 0.008~0.160 μg/ml. Moreover, excellent linear behavior over the investigated concentration range was observed, with R>0.999 for all the analytes.     

Expression, purification and characterization of a phyAm-phyCs fusion phytase

The phyA^m gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichiapastoris in order to expand the pH profile ofphytase and decrease the cost of production. The fusion phytase phyA^m-phyCs gene was successfully overexpressed in P. pastoris as an active and extracellular phytase. The yield of total extracellular fusion phytase activity is （25.4±0.53） U/ml at the flask scale and （159.1±2.92） U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 ℃ and an optimal pH at 5.5-6.0 and its relative activity remains at a relatively high level of above 70% in the range ofpH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 ℃ to 95 ℃ for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE （sodium dodecyl sulfate-polyacrylamide gel electrophoresis）. After deglycosylation by endoglycosidase H （EndoHf）, the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those ofphyCs andphyA^m.     

Anti-H IV-1 Activities of 4 Telomerase Restrictors

MTT Cell Proliferation Assay was used to optimize the concentration of Telomerase Restrictors（TRs） with minimum toxicity to the selected cells. FACSort flow cytometer and Innotest P24 HIV（Human immtmodeficiency Virus） antigen mAb ELISA Kit were used to investigate the anti-HIV-1 activities of TRs. The results showed that TRs had low cytotoxicity to the PBMC （Peripheral Blood mononuclear cells） and CEM/GFP if the concentration of TRs was at 50μmol/L or below, and the supernatant from PBMC pretreated with SHIV and TR1-001 /TR1-002 could not infect the PBMC, while can infect the C8166 with reduced infectivity, which suggested that the TRs may be one of the novel resources for screening anti-HIV-1 agents.     

Synergistically optimizing electrical and thermal transport properties of ntype PbSe

In this paper,we report that the thermoelectric performance of n-type Pb Se could be improved through synergistically optimizing electrical and thermal transport properties via Sb doping and Mg alloying.The carrier concentration was firstly optimized through Sb doping,resulting in a maximum power factor of~15.4μW cm~(-1)K~(-2)and maximum ZT of~0.9 at 873 K in Pb_(0.99)Sb_(0.01)Se.Then,Mg was selected for alloying in Pb sites to produce point defects,which can largely intensify the phonon scattering and lower thermal conductivity.After Mg alloying,the thermal conductivity at 300 K(873 K)was significantly suppressed from~4.6 Wm~(-1)K~(-1)(1.5 Wm~(-1)K~(-1))for Pb_(0.99)Sb_(0.01)Se to~2.9 Wm~(-1)K~(-1)(1.1 Wm~(-1)K~(-1))for Pb_(0.99)Sb_(0.01)Se-6%Mg Se.Through combining Sb doping and Mg alloying,a maximum ZT of~1.1 was achieved at 873 K for Pb_(0.99)Sb_(0.01)Se-6%Mg Se,and the average ZT(ZT_(ave))was increased by 28.6%from~0.42 for Pb_(0.99)Sb_(0.01)Se to~0.54 for Pb_(0.99)Sb_(0.01)Se-6%Mg Se.The results indicate that Pb Se is a robust candidate for medium-temperature thermoelectric applications.     

Polymer coated nanodiamonds as gemcitabine prodrug with enzymatic sensitivity for pancreatic cancer treatment

The fabrication of Gemcitabine (GEM) prodrug was reported to be an effective method to enhance its pancreatic cancer treatment efficiency. Here, a kind of nanocarbon-based materials, nanodiamond (ND), was selected as the nanocarrier of GEM, owing to its outstanding surface properties and non-cytotoxicity. The polyelectrolytes, polyethyleneimine and polyacrylic acid, were used to self-assemble outside ND surface through electrostatic forces, followed by attachment of polyethylene glycol to address better biocompatibility. GEM was conjugated with an enzyme-sensitive peptide gly-phe-leu-gly to build up the controlled release platform. From characterization results of dynamic laser scattering, zeta potential and transmission electron microscope, the significant improvement of ND stability in physiological condition was proved. Non-cytotoxicity of this functionalized ND carriers and cytotoxicity of the prodrug against BxPC-3 pancreatic cancer cells were indicated by methylthiazolyl tetrazolium (MTT) assay. In vivo experiments also revealed its superior anticancer effect compared with free GEM treatment. Therefore, the combination of polymer coated NDs with high surface capability and enzyme-responsive intracellular GEM release make it possible to realize higher treatment efficiency on pancreatic tumor therapy.     

Ginsenosides stimulated the proliferation of mouse spermatogonia involving activation of protein kinase C

The effect of ginsenosides on proliferation of type A spermatogonia was investigated in 7-day-old mice. Spermatogonia were characterized by c-kit expression and cell proliferation was assessed by immunocytochemical demonstration of proliferating cell nuclear antigen (PCNA). After 72-h culture, Sertoli cells formed a confluent monolayer to which numerous spermatogonial colonies attached. Spermatogonia were positive for c-kit staining and showed high proliferating activity by PCNA expression. Ginsenosides (1.0~10 μg/ml) significantly stimulated proliferation of spermatogonia. Activation of protein kinase C (PKC) elicited proliferation of spermatogonia at 10⁻⁸ to 10⁻⁷ mol/L and the PKC inhibitor H₇ inhibited this effect. Likewise, ginsenosides-stimulated spermatogonial proliferation was suppressed by combined treatment of H₇. These results indicate that the proliferating effect of ginsenosides on mouse type A spermatogonia might be mediated by a mechanism involving the PKC signal transduction pathway.     

Effects of Nd<Superscript>3+</Superscript> and Sm<Superscript>3+</Superscript> on the proliferation,differentiation and mineralization function of primary osteoblasts <Emphasis Type="Italic">in vitro</Emphasis>

A series of experimental methods including 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) test, alkaline phosphatase (ALP) activity measurement, Oil Red O stain and measurement, mineralized function expression and quantitive real time RT-PCR (qRT-PCR) were employed to assess the effect of Nd³⁺ and Sm³⁺ on the proliferation, differentiation and mineralization function of primary osteoblasts (OBs) in vitro at cell and molecular levels. The experimental results suggest that concentration, culture time and ion species are pivotal factors for switching the biological effects of rare earth ions from toxicity to activity, from damage to protection, or from down-regulation to up-regulation.     

胡桃醌含药血清体外抗肿瘤实验研究

研究胡桃醌(Juglone)含药血清对人肺癌A549细胞的生长抑制作用及对细胞周期的影响. 以血清药理学原理为基础,通过MTT法检测胡桃醌对A549细胞的生长抑制作用,并筛选最佳药物浓度;利用流式细胞仪分析胡桃醌对A549细胞周期的影响,并用Western blot验证. 实验结果表明,胡桃醌的半衰期T_1/2=136 min,MTT法显示其对体外培养的人肺癌A549细胞具有明显抑制作用,比较12,24,48 h细胞存活率得出10%高剂量组(0.107 μg/mL)胡桃醌血清抑制作用较好,同时,胡桃醌可通过凋亡途径阻滞细胞于S期,增加caspase-3的表达. 胡桃醌对A549细胞可能具有生长抑制作用,将细胞阻滞在S期,诱导细胞凋亡,上调caspase-3表达量.      

6-(1H-吲哚-3-甲基)- 5-乙基-3H-嘧啶-4-酮类化合物的合成及抗HIV活性研究

 基于二氢烷氧苄基嘧啶酮(DABO)类非核苷类逆转录酶抑制剂(NNRTIs)的构效关系研究,设计合成了2个新的6-(1H-吲哚-3-甲基)- 5-乙基-3H-嘧啶-4-酮类化合物,并采用C8166细胞进行了体外抗HIV活性测试,为新型S-DABO类非核苷类逆转录酶抑制剂结构修饰提出了新的设想.     

Association of p53 codon 72 polymorphism with liver metastases of colorectal cancers positive for p53 overexpression

Objective： To evaluate the association between p53 codon 72 polymorphism （R72P） and the risk ofcolorectal liver metastases. Methods： The p53 R72P genotype was identified by polymerase chain reaction-restriction fragment length polymorphism （PCR-RFLP） method in 78 consecutive colorectal cancer patients with liver metastases and 214 age- and sex-matched cases with nonmetastatic colorectal cancer. Results： The R allele of the p53 R72P polymorphism was more frequently found in metastatic cases than in nonmetastatic cases （P=0.075）. Carriers of the 72R allele had a 2.25-fold （95% CI （confidence interval）=1.05-4.83） increased risk of liver metastases. On the stratification analysis, 72R-carrying genotype conferred a 3.46-fold （95% CI=1.02-11.72） and a 1.05-fold （95% CI=0.36-3.08） increased risk of liver metastases for p53 overexpression-positive and negative colorectal cancers, respectively. Conclusion： These results demonstrate for the first time that the 72R allele of the p53 polymorphism has an increased risk for liver metastases in colorectal cancers positive for p53 overexpression.     

Construction of recombinant industrial Saccharomyces cerevisiae strain with bglS gene insertion into PEP4 locus by homologous recombination

The bglS gene encoding endo-1,3-1,4-β-glucanase from Bacillus subtil＆ was cloned and sequenced in this study. The bglS expression cassette, including PGK1 promoter, bglS gene fused to the signal sequence of the yeast mating pheromone a-factor （MFals）, and ADH1 terminator with G418-resistance as the selected marker, was constructed. Then one of the PEP4 allele of Saccharomyces cerevisiae WZ65 strain was replaced by bglS expression cassette using chromosomal integration of polymerase chain reaction （PCR）-mediated homologous recombination, and the bglS gene was expressed simultaneously. The recombinant strain S. cerevisiae （SC-βG） was preliminarily screened by the clearing hydrolysis zone formed after the barley β-glucan was hydrolyzed in the plate and no proteinase A （PrA） activity was measured in fermenting liquor. The results of PCR analysis of genome DNA showed that one of the PEP4 allele had been replaced and bglS gene had been inserted into the locus of PEP4 gene in recombinant strains. Different endo-1,3-1,4-β-glucanase assay methods showed that the recombinant strain SC-βG had high endo-1,3-1,4-β-glucanase expression level with the maximum of 69.3 U/（h·ml） after 60 h of incubation. Meanwhile, the Congo Red method was suitable for the determination of endo-1,3-1,4-β-glucanase activity during the actual brewing process. The current research implies that the constructed yeast strain could be utilized to improve the industrial brewing property of beer.     

Anti-allergic effects of ethanol extracts from brown seaweeds

Ethanol extracts of brown seaweeds from Pakistan and China were isolated and compared for their antiallergenic activities. They included Sargassum tennerimum (ST) and Sargassum cervicorne (SC) from Pakistan, and Sargassum graminifolium turn (SG), Sargassum thunbergii (STH), and Laminaria japonica (LJ) from China. The ethanol extracts of these brown seaweeds were optimized at 85% (v/v) ethanol for the maximum yield of phlorotannin, an inhibitor against hyaluronidase. Total phlorotannins contained in the crude extracts were measured as 1.71% (SG), 0.74% (STH), 0.97% (LJ), 3.30% (SC), and 5.06% (ST). The 50% inhibitory concentrations (IC₅₀) of Pakistani SC and ST were 109.5 and 21 μg/ml, respectively, lower than those of Chinese SG, STH, and LJ (134, 269, and 148 μg/ml, respectively). An antiallergic drug, disodium cromoglycate (DSCG), had an IC₅₀=39 μg/ml, and a natural inhibitor of hyaluronidase, catechin, had an IC₅₀=20 μg/ml. The IC₅₀ of ST extract was found similar to that of catechin (21 vs 20 μg/ml) and lower than that of DSCG (21 vs 39 μg/ml). This suggests that ST is a potent inhibitor of hyaluronidase, indicating a promising future development of natural antiallergic medicines or functional foods.     

HCV不同基因型对慢性丙型肝炎抗病毒治疗疗效的影响

目的探讨干扰素联合利巴韦林治疗慢性丙型肝炎患者临床疗效的相关影响因素.方法对110例慢性丙型肝炎患者进行PEG-IFNα-2a 180μg/周联合RBV 800~1200 mg/d的标准治疗,并完成治疗后24周随访,同时分析HCV基因型、HCV-RNA定量对持续病毒学应答(SVR)的影响.应用罗氏试剂检测丙肝病毒定量,应用反转录-套式聚合酶链反应(PT-PCR)法检测HCV基因型.结果在110例进行标准联合治疗、并完成治疗后24周随访的慢性丙型肝炎患者中,68例患者获得SVR.其中,HCV基因1b型感染者SVR率为51.4%(37/72),HCV基因N-1b型感染者SVR率为81.5%(31/38);基线HCV-RNA水平1.0×10~5IU/m L的患者SVR率为71.2%(42/59),基线HCV-RNA水平≥1.0×10~5IU/m L的患者SVR率为50.9%(26/51).结论非1b基因型SVR率高于1b基因型;基线低病毒RNA水平(血清HCV-RNA水平1.0×10~5IU/m L)SVR率高于基线高病毒RNA水平(血清HCV-RNA水平≥1.0×10~5IU/m L).     

胡椒烯丙酮的绿色合成新方法

报道了一种绿色合成胡椒烯丙酮[3160-37-0]的新方法.这种不污染环境的合成方法是用洋茉莉醛[120-57-O]与丙酮[67-64-1]在水相和有机相之间进行克莱森-施密特反应;产物分离后有机层循环再用,水层经过活性炭吸附处理后也循环使用,做到了无试验废水排放.1mol洋茉莉醛、回收的苯溶液200ml、补充丙酮70ml、回收并且经过活性炭吸附处理后的水溶液200ml、补充1.1g氢氧化钠,50℃反应7h,产率99.7%,纯度99.6%.     

Corrosion study of resorbable Ca60Mg15Zn25 bulk metallic glasses in physiological fluids

The corrosion activity of amorphous plates of Ca_(60)Mg_(15)Zn_(25)alloy was investigated.The biocompatible elements were selected for the alloy composition.The electrochemical corrosion and immersion tests were carried out in a multi-electrolyte fluid and Ringer's solution.Better corrosion behavior was observed for the samples tested in a multi-electrolyte fluid despite the active dissolution of Ca and Mg in Ringer's solution.The experimental results indicated that reducing concentration of NaCl from 8.6 g/dm~3for Ringer's solution to 5.75 g/dm~3caused the decrease of the corrosion rate.The volume of the hydrogen evolved after 480 min in Ringer's solution(40.1 ml/cm~2)was higher in comparison with that obtained in a multi-electrolyte fluid(24.4 ml/cm~2).The values of opencircuit potential(E_(OCP))for the Ca_(60)Mg_(15)Zn_(25)glass after 1 h incubation in Ringer's solution and a multielectrolyte fluid were determined to be-1553 and-1536 m V vs.a saturated calomel electrode(SCE).The electrochemical measurements indicated a shift of the corrosion current density(j_(corr))from 1062μA/cm~2for the sample tested in Ringer's solution to 788μA/cm~2for the specimen immersed in a multi-electrolyte fluid.The corrosion products analysis was conducted by using the X-ray photoelectron spectroscopy(XPS).The corrosion products were identified to be CaCO_3,Mg(OH)_2,CaO,MgO and Zn O.The mechanism of corrosion process was proposed and described based on the microscopic observations.The X-ray diffraction and Fourier transform infrared spectroscopy(FTIR)also indicated that Ca(OH)_2,CaCO_3,Zn(OH)_2and Ca(Zn(OH)_3)_2·2H_2O mainly formed on the surface of the studied alloy.     

Biosorption and biodegradation of polycyclic aromatic hydrocarbons by Phanerochaete chrysosporium in aqueous solution

Biosorption and biodegradation of phenanthrene and pyrene by live and heat-killed Phanerochaete chrysosporium are investigated to elucidate the bio-dissipation mechanisms of polycyclic aromatic hydrocarbons(PAHs) in aqueous solution and its regulating factors.The effects of nutrient conditions(carbon source and nitrogen source concentrations),the co-existing Cu 2+,and repeated-batch feed of PAHs on the biosorption and biodegradation are systematically studied.The removal of PAHs by dead bodies of P.chrysosporium is attributed to biosorption only,and the respective partition coefficients of phenanthrene and pyrene are 4040 and 17500 L/kg.Both biosorption and biodegradation contribute to the dissipation of PAHs by live P.chrysosporium in water.After a 3-d incubation,the removal percentage via biosorption are 19.71% and 52.21% for phenanthrene and pyrene,respectively.With the increase of the incubation time(3 40 d),biodegradation gradually increases from 20.40% to 60.62% for phenanthrene,and from 15.55% to 49.21% for pyrene.Correspondingly,the stored-PAHs in the fungal bodies decrease.Under the carbon-rich and nitrogen-limit nutrient conditions,the removal efficiency and biodegradation of phenanthrene and pyrene are significantly promoted,i.e.99.55% and 92.77% for phenanthrene,and 99.47% and 83.97% for pyrene after a 60-d incubation.This phenomenon is ascribed to enhanced-biosorption due to the increase of fungal biomass under carbon-rich condition,and to stimulated-biodegradation under nitrogen-limit condition.For the repeated-batch feed of phenanthrene,the pollutant is continuously removed by live P.chrysosporium,and the contribution of biodegradation is enhanced with the repeated cycles.After 3 cycles,the biodegradation percentage is up to 90% with each cycle of a 6-d incubation.