CUMULATIVE RESULTS OF CHEMOSENSITIVITY TEST USING MTT ASSAY IN DOUBLE-LAYER AGAROSE

Objective To investigate cumulative results of chemosensitivity test using 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl （MTT） assay in double-layer agarose. Methods A total of 2 491 patients with different kinds of cancers were enrolled in the study, in which 18 kinds of different anticancer drugs were used. A computer soft was used to get charts. ResultsThe total evaluability rate was 82.7% （2 060/2 491）. Among all agents, the efficiency rates of 5-Fu, MMC, DDP, BLM and CBP were higher than the efficiency rates of others. The response rate range of different cancer in vitro sensitivity by using MTT assay in double layer agarose were from 9.2% （biliary duct） to 37.5% （malignant lymphoma）. For colon and rectum cancer, 5-Fu, DDP, MMC and BLM were more sensitive than other anti-tumor agents. For breast carcinomas, ACTD and DDP were more sensitive. For gastric cancer, 5-Fu, DDP and BLM were more sensitive. For leukemia, VM-26 and HHRT were more sensitive. ACM was more sensitive to kidney and MXT and BLM were more sensitive to pancreas cancer. For Lung cancer, DDP and EPI were more sensitive. Mean true positive rate, mean true negative rate, mean sensitivity, mean specificity and mean accuracy were 44% , 92% , 72% , 77% , and 76% , respectively. Conclusion Chemosensitivity tesing using the MTT assay in a double layer agarose was a very useful reference to chem- therapy.

Heteroatom tuning in agarose derived carbon aerogel for enhanced potassium ion multiple energy storage

The incorporation of heteroatoms into carbon aerogels(CAs)can lead to structural distortions and changes in active sites due to their smaller size and electronegativity compared to pure carbon.However,the evolution of the electronic structure from single-atom doping to heteroatom codoping in CAs has not yet been thoroughly investigated,and the impact of codoping on potassium ion(K+)storage and diffusion pathways as electrode material remains unclear.In this study,experimental and theoretical simulations were conducted to demonstrate that heteroatom codoping,composed of multiple heteroatoms(O/N/B)with different properties,has the potential to improve the electrical properties and stability of CAs compared to single-atom doping.Electronic states near the Fermi level have revealed that doping with O/N/B generates a greater number of active centers on adjacent carbon atoms than doping with O and O/N atoms.As a result of synergy with enhanced wetting ability(contact angle of 9.26°)derived from amino groups and hierarchical porous structure,ON-CA has the most optimized adsorption capacity(−1.62 eV)and diffusion barrier(0.12 eV)of K^(+).The optimal pathway of K^(+)in ON-CA is along the carbon ring with N or O doping.As K^(+)storage material for supercapacitors and ion batteries,it shows an outstanding specific capacity and capacitance,electrochemical stability,and rate performance.Especially,the assembled symmetrical K^(+)supercapacitor demonstrates an energy density of 51.8 Wh kg^(−1),an ultrahigh power density of 443Wkg^(−1),and outstanding cycling stability(maintaining 83.3%after 10,000 cycles in 1M KPF6 organic electrolyte).This research provides valuable insights into the design of highperformance potassium ion storage materials.

maltosyl-agarose亲和层析法提取人肺灌洗液中肺表面活性物质结合蛋白A及其鉴定

目的:提取及纯化人肺灌洗液中肺表面活性物质结合蛋白A(SP-A)。方法:由支气管肺泡灌洗液经10000g离心40min获得的沉淀中提取SP-A。沉淀溶解于6M的尿素,SP-A蛋白重新复性后上清过maltosyl-agarose柱进行亲和层析,用梯度EDTA洗脱SP-A,并过surpose6柱纯化。聚丙烯酰胺电泳和Westernblotting鉴定。结果:经亲和层析法可以从人肺灌洗液中提取出高纯度的SP-A,聚丙烯酰胺电泳在36ku和70ku处可见清晰条带,在分子量118ku以上亦可看到3条清晰条带,经Westernblotting鉴定均为特异性SP-A蛋白。结论:采用maltosyl-agarose亲和层析法可以从肺灌洗液中获得高纯度的SP-A。为进一步研究SP-A的功能和制备SP-A单克隆抗体提供了充足的SP-A来源。

植物胞外钙调素与拮抗剂W7-Agarose结合作用的荧光光谱法研究

W7-agarose是常用的细胞外CaM功能的拮抗剂，本实验采用荧光光谱法研究了水溶液中钙调素拮抗剂W7-agarose与植物胞外钙调素的相互结合反应。W7-agarose是一种将W7-共价连接到颗粒型agamse（琼脂糖）的粒子。W7-agarose颗粒较大且容易沉淀，静置5min后，溶液中的荧光强度完全由游离的CaM产生。在溶液中加入W7-agamse后，溶液中一部分CaM与其结合后沉降至荧光比色皿底部，导致溶液中CaM的荧光强度下降。由此可以确定溶液中游离CaM的浓度。根据公式lg{[Q]t（F0-F）／F0}=nlg{[Q]tF/F0}＋lgnK[B]t，从而计算出配位体系的结合常数和配比。研究表明：二者以摩尔比1：1结合，其平衡常数为4．9×10^5。由此进一步计算了W7-agarose对胞外钙调素的拮抗率，在拮抗剂W7-agarose浓度达到15～20μmol／L时，拮抗率可达到90％以上，与文献报道的生物学体内实验结果一致，从分子水平上解释了W7-agarose与CaM的结合作用。

以硅胶和Agarose 6B为载体金属螯合亲和层析分离纯化重组人Cu,Zn-SOD

以层析硅胶Agarose 6B ,制备了Cu2 + 螯合亲和载体 ,利用合成的Cu2 + IDA 硅胶亲和柱和Cu2 + IDA Agarose 6B亲和柱分离纯化了重组人Cu ,Zn SOD ,并比较了这两 2亲和载体的纯化效果和性能。硅胶纯化SOD的比活、纯化倍数和回收率分别为 75 86U/mg·protein、6 .5倍和 86 .7% ;而Agarose 6B纯化SOD的比活、纯化倍数和回收率则分别为 6 2 2 3/mg·protein、5 .8倍和 93.0 %。 2种亲和载体 5次使用后 。

DNA recovery from agarose gels with a simple centrifuge-driven sephadex filtration

Conventional methods of DNA recovery from agarose gel generally require expensive equipment, extended elution times, or considerable handling of the sample after elution. We developed a simple protocol for a quick and effective recovery of DNA from agarose gels with good yield and quality. Using a Sephadex resin filled spin column, DNA fragments of 500 bp to 6 kb in an agarose gel slice were easily recovered by a 2 min centrifugation. The recovery efficiencies were over 40% -50% and the eluted DNA can be used directly for downstream application, such as polymerase chain reactions （PCR） and restriction enzyme digestion. This method could also be used to recover large DNA fragment （48 kb） without degradation. The use of Sephadex helps to remove small molecular impurities from agarose and it also reduces the chance of clogging the column filter caused by direct contact with agarose.

Poly-lactic acid and agarose gelatin play an active role in the recovery of spinal cord injury

Objective To investigate the role of poly-lactic acid and agarose gelatin in promoting the functional recovery of the injured spinal cord. Methods Poly-lactic acid （PLA） or agarose was embedded in the space between two stumps of the hemisectioned spinal cord. Immunohistochemistry was used to show astroglia proliferation and the infiltration of RhoA-positive cells. Locomotor activity recovery was evaluated by testing the function of hindlimbs. Results Astroglias and RhoA labeled non-neuronal cells accumulated in the area adjacent to the implant, while the number of RhoA-posirive cells was decreased dramatically in the absence of implant. Animals implanted with agarose gelatin recovered more quickly than those with PLA, concomitant with a higher survival rate of the neurons. Conclusion Both PLA and agarose gelatin benefited the recovery of spinal cord after injury by providing a scaffold for astroglia processes. Modulation of the rigidity, pore size and inner structure of PLA and agarose gelatin might make these biodegradable materials more effective in the regeneration of the central nervous system （CNS）.

Efficient Extraction of Agarose from Red Algae Using Ionic Liquids

We explored the possibility of using ionic liquids (ILs) as medium for efficient extraction of agarose via dissolution of red algae under varying conditions of heating or microwave irradiation. As compared to conventional methods, a very high extraction yield of good quality agarose (as high as 39 wt%) could be achieved depending upon the nature of used IL and applied experimental conditions. Purity of extracted agarose was confirmed from various spectral and analytical techniques, such as 1H and 13C NMR, FTIR, circular dichroism (CD), gel permeation chromatography (GPC) and thermogravimetric analysis (TGA). The physicochemical properties, such as gelling or melting temperature, viscosity and gel strength of extracted agarose hydrogels have been measured and compared with the agarose obtained from similar source reported in the literature. ILs were recovered after the extraction of agarose and were reused for further extraction experiments. % Recycling and extraction ability of recycled ILs in different cycles have been measured. The developed extraction process of utilizing ILs as medium is easy, simple and highly efficient as compared to the conventional methods of agarose extraction from algae.

Fabrication of agarose hydrogel with patterned silver nanowiresfor motion sensor

In this work, a facile strategy is proposed to construct stretchable electronics based on agarose hydrogels. The hot agarose solution is casted onto a template with patterned Ag nanowires, endowing agarose hydrogel with patterned conductive surface. After further heating treatment, Ag nanowires can be embedded into the agarose hydrogel, which improves the stability of Ag pattern and has no obvious e ffect on the conductivity of hydrogels. The agarose hydrogel with patterned Ag nanowires is certi fied to be an e ffective stretchable electrode to record the motion of joints, which has great potential applications in the field of wearable devices.

Downregulation of iASPP Expression Suppresses Proliferation, Invasion and Increases Chemosensitivity to Paclitaxel of Head and Neck Squamous Cell Carcinoma In Vitro

Objective Our previous study has revealed that iASPP is elevated in human head and neck squamous cell carcinoma(HNSCC)and iASPP overexpression signifcantly correlates with tumor malignant progression and poor survival of HNSCC.This study investigated the function of iASPP playing in proliferation and invasion of HNSCC in vitro.Methods HNSCC cell line Tu686 transfected with Lentiviral vector-mediated iASPP-specific shRNA and control shRNA were named the shRNA-iASPP group and shRNA-NC group,respectively.The non-infected Tu686 cells were named the CON group.CCK-8 assay,flow cytometry,transwell invasion assay were performed to detect the effects of iASPP inhibition in vitro.Results Our results demonstrated that the proliferation of shRNA-iASPP cells at the time of 72 h(F=32.459,P=0.000),96 h(F=51.407,P=0.000),120 h(F=35.125,P=0.000)post-transfection,was significantly lower than that of shRNANC cells and CON cells.The apoptosis ratio of shRNA-iASPP cells was 9.42%±0.39%(F=299.490,P=0.000),which was significantly higher than that of CON cells(2.80%±0.42%)and shRNA-NC cells(3.18%±0.28%).The percentage of shRNA-iASPP cells in G0/G1 phase was 74.65%±1.09%(F=388.901,P=0.000),which was strikingly increased,compared with that of CON cells(55.19%±1.02%)and shRNA-NC cells(54.62%±0.88%).The number of invading cells was 56±4 in the shRNA-iASPP group(F=84.965,P=0.000),which decreased significantly,compared with the CON group(111±3)and the shRNA-NC group(105±8).The survival rate of shRNA-iASPP cells administrated with paclitaxel was highly decreased,compared with CON cells and shRNA-NC cells(F=634.841,P=0.000).Conclusion These results suggest iASPP may play an important role in progression and aggressive behavior of HNSCC and may be an efficient chemotherapeutic target for the treatment of HNSCC.

Preparation and characterization of agar, agarose,and agaropectin from the red alga Ahnfeltia plicata

Agar,agarose,and agaropectin were extracted from the red alga Ahnfeltia plicata,and their properties and structures were characterized.Agar was extracted by a comparatively low alkaline consumption of 1.2%.It exhibited a gel strength of 1 152.50±74.25 g/cm^2 and a sulfate content of 0.55%±0.08%.The yield of agar from A.plicata was 24.53%,which is higher than those of other agarophytes commonly used in China.Three kinds of the method were compared for the purification of agarose,and the physicochemical properties of agarose that was prepared under the optimal condition were identical to those of commercially available agarose.Furthermore,agaropectin was purified from A.plicata and characterized by GC,HPLC,UV-spectrum,and FI-IR to understand its composition and structure.It was the first time to comprehensively study the agar and its fractions from the red alga of A.plicata.This research provided an eco-friendly agar extraction method from A.plicata and revealed its potential application for the production of agar,agarose,and agaropectin.

Searching for the best agarose candidate from genus Gracilaria, Eucheuma,Gelidium and local brands

Objective: To explore the potential of local agar of genus Gracilaria, Eucheuma,Gelidium and local brands as an alternative for imported agarose for DNA electrophoresis, and to examine their ability related to separation and migration of DNA fragments in DNA electrophoresis.Methods: Their performance at various concentrations were compared via an experimental study with a specific brand of imported commercial agarose used in molecular biology research. The measured variables were separation and migration during electrophoresis of a DNA fragment.Results: The local agar genus Gracilaria gigas, Gelidium, brand “B” and brand “S”could separate DNA fragments at a concentration between 1% and 2%, with an optimum concentration of 2% w/v, as good as a specific brand of imported commercial agarose.Conclusions: Their performance were very close to that of commercial agarose and can still be improved by further agar purification as well as by p H and sulfur control.

AFFINITY CHROMATOGRAPHY PURIFICATION OF UROKINASE WITH EPICHLOROHYDRIN ACTIVATED AGAROSE MATRIX

1 INTRODUCTIONIn literature,most matrices of affinity chromatography for urokinase(EC 3.4.99.26)purification were prepared by cyanogen bromide activation.However,theseadsorbents usually suffered from the drawback of leakage of the ligand,particularly inalkaline medium,because of the instability of the isourea linkage between the ligandand the spacer or agarose.Moreover,the positively charged imido group of theN-substituted isourea derivative and the hydrophobicity of the spacers might promotenonspecific adsorption.On the contrary,the adsorbents prepared by the method

Application value of ATP based bioluminescence tumor chemosensitivity assay in the chemotherapy for hydrothorax caused by non-small cell lung cancer

Objective： The aim of the study was to investigate the clinical value and application of ATP based bioluminescence tumor chemosensitivity assay （ATP-TCA） in the chemotherapy for hydrothorax caused by non-small cell lung cancer （NSCLC）. Methods： Hydrothorax specimens from 120 NSCLC patients were analyzed by ATP-TCA and the most sensitive chemotherapeutic drugs were used in NSCLC patients （treatment group）. At the same time, 56 NSCLC patients with hydrethorax were admitted in our Hospital （Department of Oncology, The No. 2 People＇s Hospital of Yibin, China） and given chemotherapy without guidance of the ATP-TCA （control group）. Before the third chemotherapeutic cycle, clinical outcomes were analyzed in the two groups. Results： Effective rate of hydrothorax in treatment group was 67%, while 46% in control group （P 〈 0.05）. In refractory hydrothorax patients, they were 69% and 40% （P 〈 0.05）, respectively.In vitro results correlated well with clinical outcomes （P 〈 0.01）. Conclusion： Effective rate of chemotherapy for hydrothorax in NSCLC is higher in treatment group than that in control group. ATP-TCA is especially helpful for refractory hydrothorax.

Effects of agarose concentration and addition on the properties of gel-cast 3Y-ZrO2 green and sintered bodies

Using agarose as a gel agent to prepare ceramics is a suitable non-toxic gel-casting method. The effects of agarose concentration and addition amount on gel-cast 3Y-ZrO2 green and sintered bodies were investigated. Green bodies were prepared by gel-casting using an agarose solution of a different agarose mass fraction including 3.0%, 3.5%, 4.0%, 4.5%, 5.0% and 5.5% with an added amount of agarose in the green body at a mass fraction of 0.7%, and using the agarose solution of a mass fraction of 4.5% with a different amount of added agarose including 0.6%, 0.7%, 0.8%, 0.9%, and 1.0% in terms of mass fraction. The green bodies were sintered at I 600 ~C for 4 h. The relative density, linear shrinkage, and bending strength of the green bodies, and L^e density, microstructure, bending strength, and fracture toughness of sintered samples were characterized and analyzed. Results show that comprehensive properties of green bodies made with 4.5% （in terms of mass fraction） agarose solution added to an mass fraction of agarose in the slurry at 0.6% were the best, of which the bending strength was 3.14 MPa, the linear shrinkage was below 4.3%, and the relative density is 57.3%. For sintered bodies, the best fracture toughness was 5.63 MPa m1/2, and the optimal density was 5.88 gcm-3.

A Rapid Electrophoresis Method on Agarose Gel to Characterise Dairy Protein Aggregates

Heat treatment of milk may cause whey proteins and caseins to form aggregates. These soluble and micellar aggregates and their other properties (size, composition, shape, etc.) can affect the techno-functionalities to the milk, conferring interesting or negative features depending on the application in dairy industries. In this study, we propose a new approach to characterise those protein aggregates. SDS-agarose electrophoresis is followed by the calculation of a retention factor (Rf) for each protein spot. Rf allows milk aggregates to be compared qualitatively under the same conditions. This method could be transposed to the dairy industry for a better knowledge of the milk subsequent to heat treatment.

Agarose hydrogel doped with soluble Se-chitosan for Se-enriched cultivation of sprouts

Se-chitosan with a high Se content of 139 mg/g was successfully synthesized using the glacial acetic acid-sodium selenite(HAc-Na2SeO3)method.The chemical structure of Se-chitosan was characterized by FTIR,XRD,TGA,UV–vis and 13C NMR,confirming that selenite acid was introduced into chitosan via the esterification reaction of the hydroxyl group at C6 and the electrostatic interaction of the amino group at C2 of chitosan.Agarose hydrogels were designed as an efficient carrier of Se-chitosan for Se-enriched cultivation of sprouts.The hydrogels exhibited a porous structure,which is beneficial for Se diffusion in hydrogels.The growth indexes of sprouts grown in Agar/Se-chitosan composite hydrogels with different Se concentrations,including plant weight,plant height and Se content,were investigated.The results showed that in the Se-enriched cultivation of sprouts,Se-chitosan possessed an excellent Se enrichment effect.As the Se concentration in Agar/Se-chitosan composite hydrogels increases from 0 to 1600μg/L,the Se content of the sprouts will also increase from 0.92 mg/kg to 6.79 mg/kg accordingly.Moreover,it is necessary to control the amount of Se-chitosan added to the hydrogels;otherwise,the growth of sprouts will be inhibited.This research provided a simple and effective method for the fabrication of Secontaining hydrogels for Se-enriched cultivation of sprouts.

GLUTATHIONE S TRANSFERASE-π EXPRESSION IN HU- MAN GASTRIC CANCER AND ITS RELATIONSHIP TO CHEMOSENSITIVITY

Objective To investigate the expression of glutathione S transferase π(GST-π) in gastric carcinoma tissue sections,and its relationship to the clinicopathological parameters and the results of chemosensitivity testing on the freshly purified human gastric cancer cells. Methods Expression of GST-π in 39 gastric cancer specimens was detected by immunohistochemical method.Simultaneously,the chemosensitivity of individual cases to 5-FU,CDDP,MMC and ADM was determined by using MTT colorimetric assay. Results The percentage of positive GST-π was 66.7%,while that of negative was 33.3%.No significant correlation between the expression of GST-π and clinicopathologic features was observed.There was a statistical significance of the correlation of GST-π expression with drug resistance of gastric cancer to CDDP and MMC. Conclusion Over-expression of GST-π in gastric cancer tissue may be related to the drug resistance to CDDP and MMC.It is therefore important to select carefully the optimal anticancer drug for patients with highly positive GST-π expression.

Applying Both Chemical Liquefaction and Enzymatic Catalysis Can Increase Production of Agaro-Oligosaccharides from Agarose

Red algae represents an important marine bioresource.One high-value utilization of red algae is the production of agarooligosaccharides which have many positive biological effects.However,the lack of an efficient production route seriously limits the application of agaro-oligosaccharides.In this study,we established a green route that combines chemical liquefaction and enzymatic catalysis for the efficient production of agaro-oligosaccharides,and we used the production of neoagarotetraose(NA4)as an example.Agarose(150 g L^−1)liquefaction by citric acid was controlled with respect to two targets:a 100%liquefaction rate and a high average degree of polymerization(>4)of the liquesced agaro-oligosaccharides,which were then catalyzed byβ-agarase into an oligosaccharides mixture with a high concentration of NA4(30.8 g L^−1)in a 1-L reaction volume.After purification,we obtained 25.5 g of NA4 with a purity of 92%.This work establishes an easy route for the efficient production of pure agaro-oligosaccharides from agarose.

The preparation of low electroendosmosis agarose and its physico-chemical property

Studies on Gelidium amansii agar fractionations were carried out in this paper. Gelidium amansii agar was fractionated on DEAE-Cellulose, and four fractions were obtained sequentially. The fractions were analyzed on physical and chemical properties, and IR and 13C-NMR spectroscopy applied for elucidating the chemical structure. Among the four fractions obtained, water fraction measured up to the standard of low EEO agarose. The sulfate content, ash content, electroendosmosis and gel strength (1%) of water fraction were 0.16%, 0.34%, 0.12 and 1 130g/cm2 respectively, similar to those of the Sigma products.