Optimization of SSR-PCR Non-denatured Polyacrylamide Gel Electrophoresis Conditions in Kernelled Apricot

[Objective] The aim was to optimize the SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot.[Method]25 accessions of kernelled apricot and three accessions of edible apricot were selected as experimental materials to screen the repeatable SSR loci with high polymorphism by the use of SSR markers combined with non-denatured polyacrylamide gel electrophoresis.And the effect of different factors on electrophoresis conditions was compared to explore the optimal SSR-PCR non-denatured polyacrylamide gel electrophoresis conditions in kernelled apricot.[Result]The optimal non-denatured polyacrylamide gel electrophoresis conditions for SSR-PCR were established as follows：polyacrylamide gel concentration 6%,the ratio of acrylamide to bisacrylamide 29∶1,electrophoresis at 1 000 V for 2-3 h,and staining for 15 min within 0.1% AgNO3.[Conclusion]The optimum electrophoresis system has provided some technical foundations to further study the phylogenetic relationship of kernelled apricots by SSR markers.

Improvement of Polyacrylamide Gel Electrophorus in Phytophthora infeatans SSR Marker

[Objective] The paper was to improve polyacrylamide gel electrophorus in Phytophthora infeatans SSR Marker.[Method] With the disease sample of P.infeatans collected from Guyuan in Ningxia Province in 2009 as test material,its DNA was extracted and amplified with PCR,and its products were carried out polyacrylamide gel electrophoresis.[Result] 12% polyacrylamide gel electrophoresis was used to detect primers D13,G11 and PI02,and 8% polyacrylamide gel electrophoresis was used to detect primers PI4B,PI63,SSR4,SSR8 and SSR11,then 0.1% silver nitrate was used to stain,and an ideal electrophoresis and staining effect was obtained.[Conclusion] The electrophoresis and staining method suitable for P.infeatans SSR Marker established in the study had the characteristics of high sensitivity,simple operation and clear bands,which was an effective,simple and quick detection method.

Synthesis of YAG∶Ce^(3+) Phosphor by Polyacrylamide Gel Method and Promoting Action of α-Al_2O_3 Seed Crystal on Phase Formation

YAG： Ce^3 ＋ phosphor particles were prepared using polyacrylamide gel method. The structure evolution of powders during annealing process was followed by X-ray diffraction determination. It is found that some intermediate phases, including θ-Al2O3, YAM and YAP, are formed when calcining polyacrylamide gel, however, the pure YAG phase can be formed directly when calcining polyacrylamide gel with α-Al2O3 as seed crystal. These facts show that the existence of α- Al2O3 seed crystal can block the formation of θ-Al2O3, YAM and YAP, and accelerate its reaction with Y2O3 to form YAG phase directly at lower temperature. The emission peak of prepared YAG ： Ce^3 ＋ phosphor is wide with maximum at 550 nm and the exitation band has two peaks, the major one is around at 460 nm, which matches the blue emission of GaN LED and is suitable for the assemble of white LED. Some fluxes can enhance the photoluminescence intensity of phosphor particles, that can be attributed both to the improvement of crystallization processes of YAG and to the stabilization of trivalence cerium ion in YAG：Ce^3 ＋.

Synthesis,Characterization and Electromagnetic Studies on Nanocrystalline Nickel Zinc Ferrite by Polyacrylamide Gel

Nanocrystalline nickel zinc ferrite powders （Ni=Zn1-xFe2O4, A for x=0, B for x=0.2, C for x=0.5, D for x= 0.8 and E for x= 1） were synthesized by polyacrylamide gel method. X-ray diffraction （XRD）, transmission electron microscopy （TEM） and wave-guide were used to characterize the composition. The XRD results show that the dried gel powders are amorphous, and the characteristic peaks of the spinel Ni0.5Zn0.5Fe2O4 appear after the gel is calcined at 400℃ for 1 h. When the calcining temperatures are 600 and 800℃, the average grain sizes are identified by TEM to be 10 and 30 nm, respectively. The NixZn1-xFe2O4 powders have both dielectric loss and magnetic loss in the frequency range of 8.2-11.0GHz. With the increase of Ni^2＋ ions content, the dielectric parameters （ε′） and permeability （u′） of the NixZn1-xFe2O4 powders decrease while the dielectric loss （ε″）, magnetic loss （u″） and the reflection loss increase.

Synthesis,Characterization and Electromagnetic Studies on Nanocrystalline Co_(0.5)Zn_(0.5)Fe_2O_4 Synthesized by Polyacrylamide Gel

Nanocrystalline Co0.5Zn0.5Fe2O4 ferrite was synthesized by polyacrylamide gel method. The electromagnetic and microwave absorption properties of the ferrite were investigated. The results indicated that calcining temperature of the ferrite had a significant influence on the effective properties of the ferrite. When the calcining temperature was 500, 600 and 700℃, the average size of particles was 10, 30 and 80 nm, respectively. The dielectric loss （ε″） and magnetic loss （μ″） of the ferrite was around 0.65 and 0.29 at 8.2 GHz, respectively. Microwave absorption properties of the ferrites were simultaneously influenced due to the strong correlation between reflection loss and electromagnetic parameters of the ferrite.

Characterization of YSZ Ceramic Nanopowders Synthesized at Different Temperatures via Polyacrylamide Gel Method

Tetragonal zirconia (T-ZrO2) ceramic nanopowders stabilized with 3 mol% Y2O3 were synthesized via polyacrylamide gel method, using ZrOCl2?8H2O and Y(NO3)3?6H2O as raw materials. The effect of temperature on phase composition and morphology of YSZ nanopowders and sintering behavior of YSZ ceramics was investigated by X-ray diffraction (XRD), scanning electron microscopy (SEM) and Vickers hardness tester. The aging-resistance of YSZ ceramics was measured by means of aging experiments. The results demonstrated that the phase composition of YSZ ceramic nanopowders had no obvious change and it was composed of T-ZrO2. Particle size of well-dispersed YSZ ceramic nanopowders increased from 17 to 35 nm with increasing calcining temperature from 600 to 800 ℃. There was noticeable negative correlation between calcining temperature and the relative density of YSZ ceramic at the same sintering temperature. The aging experiments showed that water vapor facilitated tetragonal to monoclinic phase transformation, and the sample that had smaller grain size exhibited better aging-resistance. It can be concluded that when the calcining temperature is 600 ℃ and sintering temperature is 1550 ℃, the relative density and hardness of YSZ ceramic arrive at the peak of 96.64% and 11.135 GPa respectively, and it has less microcracks and excellent aging-resistance.

Breast Fibromatosis after Hydrophilic Polyacrylamide Gel Injection for Breast Augmentation: a Case Report and Review of the Literature

BREAST fibromatosis is a rare kind of lesion. The average incidence is about 2-4 per million every year} So far there have been about 100 cases reported altogether.2 In this report, we describe a case of breast fibromatosis developed after hydrophilic polyacrylamide gel （HPG） injection for breast augmentation. By reviewing the literature, the possible pathogenesis of this case and the proper treatment strategy are investigated.

Isolation and purification of guinea pig inner ear antigens by preparative polyacrylamide gel electrophoresis

Objective To isolate and purify the subcomponents of guinea pig inner ear antigens for further study on the autoimmunity of the inner ear.Methods Inner ear tissues were homogenized in phosphate buffered saline containing 0.1% SDS, and then frozen and defrosted repeatedly to extract inner ear antigens. Preparative polyacrylamide gel electrophoresis was used to separate the subcomponents of inner ear antigens. Following electrophoresis,the protein bands were localized by rapid staining and destaining.Results The major protein bands were clearly distinct when 3 mg of crude inner ear antigens was loaded,and the three major subcomponents (31, 42- 45 and 60 kD proteins) accounted for about 25.99%,21.91% and 21.10%, respectively.Conclusion Preparative polyacrylamide gel electrophoresis can be used to purify the major subcomponents of inner ear antigens.

Magnetic resonance imaging on complications of breast augmentation with injected hydrophilic polyacrylamide gel

The injection augmentation mammaplasty for cosmetic purpose has been popular recently in China. Two kinds of injectable material are used clinically, autologous fat and biomaterial. The fat injection for breast augmentation is in question with the major problems of progressive fat re-absorption, microcalcification, and fat liquefaction. Now, the principal alloplastic biomaterial for injection augmentation mammaplasty in China is hydrophilic polyacrylamide gel （HPAAG）. Although thousands of breasts have been augmented with HPAAG and it seems to be a good biocompatible material, some complications develop after HPAAG injection augmentation mammaplasty.The patients had to undergo surgery to remove the injected HPAAG and associated lesions. Ultrasonography or magnetic resonance imaging （MRI） should be taken preoperatively to demonstrate the distribution of injected HPAAG and associated lesions. In this report, the diagnostic value and clinical significance of MRI on the complications of HPAAG breast augmentation were discussed.

A green method of staining DNA in polyacrylamide gel electrophoresis based on fluorescent copper nanoclusters synthesized in situ

The safety of nucleic acid staining dyes has long been recognized to be a problem. Extensive efforts have been made to search for alternatives to the most popular but toxic staining dye, ethidium bromide （EtBr）. However, so far no staining method that can be guaranteed to be suffidently safe has been developed. In this paper, we report a green staining method of DNA in polyacrylamide gel electrophoresis, where in situ synthesis of DNA-templated fluorescent copper nanoclusters （CuNCs） in the gel is achieved to make the DNA bands visible under UV light. Moreover, a comprehensive study of the performance of this staining method has been conducted and the experimental results show that it has favorable sensitivity, stability, and usability. Meanwhile, in our animal experiments, the two reagents （copper sulfate and ascorbic acid） as well as the synthesized CuNCs have been proven to be non-toxic in contact with skin. In addition, all the reagents employed in this work are readily available and low cost, and the procedure is simple to carry out. Therefore, this novel staining method based on the in situ synthesis DNA-templated fluorescent CuNCs has many potential applications.

Conformer Analysis of a Biological G-Quadruplex Element in the Human c-MYC Promoter by Native Polyacrylamide Gel Electrophoresis

The G-quadruplexes undergo complex folding and conformation exchanges.G-quadruplex stability is substantially influenced by sequence,bufer and temperature.Mutational analysis together with nuclear magnetic resonance spectroscopy(NMR),X-ray crystallography and circular dichroism spectroscopy has been proved to be a powerful approach for G-quadruplex structural analysis.Herein,we used DNA sequence mutation and native polyacrylamide gel electrophoresis to investigate the topology and conformations of a G-quadruplex model molecule Pu18 found in the human c-MYC promoter.The guanines(G6,G9 or G18)which were not contributable to G-tetrad formation in c-MYC Pu18 sequence were mutated to thymine or adenine.We screened the buffer and temperature of gel electrophoresis for Pu18.Gel electrophoresis showed that two of the four conformers of c-MYC Pu18 in 100 mM K+buffer were resolved,which was in accordance with the conformations as determined by the 1 H NMR spectra in previous studies.This technique is expected as a general methodology for its easy operation and low cost to facilitate uncovering more yet unidentified G-quadruplex folds and functions,with the assistance of other analytical methods like NMR,X-ray crystallography and circular dichroism spectroscopy.

A method to determine Young's modulus of soft gels for cell adhesion

A convenient technique is reported in this note for measuring elastic modulus of extremely soft material for cellular adhesion. Specimens of bending cylinder under gravity are used to avoid contact problem between testing device and sample, and a beam model is presented for evaluating the curvatures of gel beams with large elastic deformation. A self-adaptive algorithm is also proposed to search for the best estimation of gels＇ elastic moduli by comparing the experimental bending curvatures with those computed from the beam model with preestimated moduli. Application to the measurement of the property of polyacrylamide gels indi- cates that the material compliance varies with the concentrations of bis-acrylamide, and the gels become softer after being immersed in a culture medium for a period of time, no matter to what extent they are polymerized.

Non-decoloured In-gel Digestion of Coomassie Blue-stained Proteins

A simplified method is presented for tryptic digestion of Coomassie brilliant blue(CBB)-stained proteins in polyacrylamide gels. Compared with conventional methods, the proposed method does not require a removal of the dye before digestion, and is thus faster and saves a lot of labor. The resulted digest can be analyzed by either RPLC/ESIMS or MALDI MS for identification of the protein in a conventional way. Model studies with bovine serum albumin(BSA) showed that 50 ng of the protein could be routinely identified. The simplified procedure displays a tendency to produce more incompletely cleaved peptides, which is favorable for improving the sequence coverage.

Plant Proteomics in the Post-genomic Era

Proteomics is one of the most active research fields in the post-genomic era. Here we briefly introduce the scientific background of the origination of proteomics and its content, research method. The new developments of proteomics at the levels of individual plants, tissues, organs and organells, as well as its applications in the area of plant genetic diversity, mutant characterization, and plant physiology, etc are reviewed. At last, the challenge and prospect of proteomics are discussed.

Research on Sex and Tissue Differences in Isozymic Phenotype of Varicorhinus macrolepis through Isozyme Electrophoresis

Polyacrylamide gel electrophoresis (PAGE) and biochemical staining method were used in this study for the analysis on malate dehydrogenase (MDH,E.C. 1.1.1.37) isozymes zymogram in 11 different types of tissues of male and female Varicorhinus macrolepis. It had been found for the first time that the phenotype of malate dehydrogenase (MDH),acid phosphatase (ACP) and superoxide dismutase (SOD) showed difference between male and female V. macrolepis,and there was no difference among different individuals in the same sex. Therefore,the electrophoresis band of malate dehydrogenase,acid phosphatase and superoxide dismutase could be used as an indicator for the identification of gender and tissues of V. macrolepis,which would provide basic data for the developmental genetics,variety improvement and directed breeding of V. macrolepis groups,thus facilitating the development and protection of this valuable fish species.

Optimization of Extraction Methods of ROP GTPase from Young Leaves of Wheat

All of the Rho GTPase seems to form a special sub-family,because such a sub-family has so far only found in plants,and then named Rop GTPase,which directly involved in and regulated of muscle actin cytoskeletal reorganization,such as a series of signal transductions.The efficient purification technology and the means of ROP GTPase in wheat are the key basis of the studies on its functions and prosperities.And it has a very important theoretical and practical significance in the signal transduction and F-act...

Virulence and potential pathogenicity of coccoid Helicobacter pylori induced by antibiotics

AIM: To explore the virulence and the potential pathogenicity of coccoid Helicobacter pylori (H. pylori) transformed from spiral form by exposure to antibiotic. METHODS: Three strains of H. pylori, isolated from gastric biopsy specimens of confirmed peptic ulcer, were converted from spiral into coccoid from by exposure to metronidazole. Both spiral and coccoid form of H. pylori were tested for the urease activity, the adherence to Hep-2 cells and the vacuolating cytotoxicity to Hela cells, and the differences of the protein were analysed by SDS-PAGE and Western blot. The mutation of the genes including ureA, ureB,hpaA, vacA and cagA, related with virulence, was detected by means of PCR and PCR-SSCP. RESULTS: In the coccoid H. pylori,the urease activity, the adherence to Hep-2 cells and the vacuolating cytotoxicity to Hela cells all decreased. In strain F44, the rate and index of adherence reduced from 70.0% +/- 5.3% to 33% +/- 5.1% and from 2.6 +/- 0.4 to 0.96 +/- 0.3 (P 【 0.01), respectively. The invasion of coccoid H. pylori into Hep-2 cell could be seen under electronmicroscope. SDS-PAGE showed that the content of the protein with the molecular weight over Mr 74000 decreased, and the hybriditional signal in band M(r) 125000 weakened, while the band M(r)110000 and M(r)63000 strengthened in coccoid H.pylori as shown in Western blot. The results of PCR were all positive, and PCR-SSCP indicated that there may exist the point mutation in gene hpaA or vacA. CONCLUSION: The virulence and the proteins with molecular weight over M(r)74000 in coccoid H.pylori decrease, but no deletion exists in amplification fragments from ureA, ureB, hpaA, vacA and cagA genes, suggesting that coccoid H.pylori may have potential pathogenicity.

Comparative proteomics analysis of human gastric cancer

AIM: To isolate and identify differentially expressed proteins between cancer and normal tissues of gastric cancer by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). METHODS: Soluble fraction proteins of gastric cancer tissues and paired normal tissues were separated by 2-DE. The differentially expressed proteins were selected and identified by MALDI-TOF-MS and database search. RESULTS: 2-DE profiles with high resolution and reproducibility were obtained. Twenty-three protein spots were excised from sliver staining gel and digested in gel by trypsin, in which fifteen protein spots were identified successfully. Among the identified proteins, there were ten over-expressed and five under-expressed proteins in stomach cancer tissues compared with normal tissues. CONCLUSION: In this study, the well-resolved, reproducible 2-DE patterns of human gastric cancer tissue and paired normal tissue were established and optimized and certain differentially-expressed proteins were identified. The combined use of 2-DE and MS provides an effective approach to screen for potential tumor markers.

Recombinant scorpion insectotoxin AaIT kills specifically insect cells but not human cells

The nucleotide sequence deduced from the amino acid sequence of the scorpion insectotoxin AaIT was chemically synthesized and was expressed in Escherichia coli. The authenticity of this in vitro expressed peptide was confirmed by N-terminal peptide sequencing. Two groups of bioassays, artificial diet incorporation assay and contact insecticidal effect assay, were carried out separately to verify the toxicity of this recombinant toxin. At the end of a 24 h experimental period, more than 60% of the testing diamondback moth (Plutella xylostella) larvae were killed in both groups with LC50 value of 18.4 microM and 0.70 microM respectively. Cytotoxicity assay using cultured Sf9 insect cells and MCF-7 human cells demonstrated that the toxin AaIT had specific toxicity against insect cells but not human cells. Only 0.13 microM recombinant toxin was needed to kill 50% of cultured insect cells while as much as 1.3 microM toxin had absolutely no effect on human cells. Insect cells produced obvious intrusions from their plasma membrane before broken up. We infer that toxin AaIT bind to a putative sodium channel in these insect cells and open the channel persistently, which would result in Na+ influx and finally cause destruction of insect cells.