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Biotechnology
Differentiation of smooth muscle progenitor cells in peripheral blood and its application in tissue engineered blood vessels
Shang-zhe XIE, Ning-tao FANG, Shui LIU, Ping ZHOU, Yi ZHANG, Song-mei WANG, Hong-yang GAO, Luan-feng PAN
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 923-930.   https://doi.org/10.1631/jzus.B0820257
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Background: A major shortcoming in tissue engineered blood vessels (TEBVs) is the lack of healthy and easily attainable smooth muscle cells (SMCs). Smooth muscle progenitor cells (SPCs), especially from peripheral blood, may offer an alternative cell source for tissue engineering involving a less invasive harvesting technique. Methods: SPCs were isolated from 5-ml fresh rat peripheral blood by density-gradient centrifugation and cultured for 3 weeks in endothelial growth medium-2-MV (EGM-2-MV) medium containing platelet-derived growth factor-BB (PDGF BB). Before seeded on the synthesized scaffold, SPC-derived smooth muscle outgrowth cell (SOC) phenotypes were assessed by immuno-fluorescent staining, Western blot analysis, and reverse transcription polymerase chain reaction (RT-PCR). The cells were seeded onto the silk fibroin-modified poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (SF-PHBHHx) scaffolds by 6×104 cells/cm2 and cultured under the static condition for 3 weeks. The growth and proliferation of the seeded cells on the scaffold were analyzed by 3-(4,5-dimethylthiazol-2-yl)-diphenyltetrazolium bromide (MTT) assay, scanning electron microscope (SEM), and 4,6-diamidino-2-phenylindole (DAPI) staining. Results: SOCs displayed specific “hill and valley” morphology, expressed the specific markers of the SMC lineage: smooth muscle (SM) α-actin, calponin and smooth muscle myosin heavy chain (SM MHC) at protein and messenger ribonucleic acid (mRNA) levels. RT-PCR results demonstrate that SOCs also expressed smooth muscle protein 22α (SM22α), a contractile protein, and extracellular matrix components elastin and matrix Gla protein (MGP), as well as vascular endothelial growth factor (VEGF). After seeded on the SF-PHBHHx scaffold, the cells showed excellent metabolic activity and proliferation. Conclusion: SPCs isolated from peripheral blood can be differentiated into the SMCs in vitro and have an impressive growth potential in the biodegradable synthesized scaffold. Thus, SPCs may be a promising cell source for constructing TEBVs.
Influence of outliers on QTL mapping for complex traits
Yousaf HAYAT, Jian YANG, Hai-ming XU, Jun ZHU
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 931-937.   https://doi.org/10.1631/jzus.B0820045
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A method was proposed for the detection of outliers and influential observations in the framework of a mixed linear model, prior to the quantitative trait locus (QTL) mapping analysis. We investigated the impact of outliers on QTL mapping for complex traits in a mouse BXD population, and observed that the dropping of outliers could provide the evidence of additional QTL and epistatic loci affecting the 1stBrain-OB and the 2ndBrain-OB in a cross of the abovementioned population. The results could also reveal a remarkable increase in estimating heritabilities of QTL in the absence of outliers. In addition, simulations were conducted to investigate the detection powers and false discovery rates (FDRs) of QTLs in the presence and absence of outliers. The results suggested that the presence of a small proportion of outliers could increase the FDR and hence decrease the detection power of QTLs. A drastic increase could be obtained in the estimates of standard errors for position, additive and additive× environment interaction effects of QTLs in the presence of outliers.
QTL analysis of flag leaf in barley (Hordeum vulgare L.) for morphological traits and chlorophyll content
Da-wei XUE, Ming-can CHEN, Mei-xue ZHOU, Song CHEN, Ying MAO, Guo-ping ZHANG
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 938-943.   https://doi.org/10.1631/jzus.B0820105
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To understand genetic patterns of the morphological and physiological traits in flag leaf of barley, a double haploid (DH) population derived from the parents Yerong and Franklin was used to determine quantitative trait loci (QTL) controlling length, width, length/width, and chlorophyll content of flag leaves. A total of 9 QTLs showing significantly additive effect were detected in 8 intervals on 5 chromosomes. The variation of individual QTL ranged from 1.9% to 20.2%. For chlorophyll content expressed as SPAD value, 4 QTLs were identified on chromosomes 2H, 3H and 6H; for leaf length and width, 2 QTLs located on chromosomes 5H and 7H, and 2 QTLs located on chromosome 5H were detected; and for length/width, 1 QTL was detected on chromosome 7H. The identification of these QTLs associated with the properties of flag leaf is useful for barley improvement in breeding programs.
Functional characterization of a potassium transporter gene NrHAK1 in Nicotiana rustica
Zhao-kui GUO, Qian YANG, Xiu-qing WAN, Pei-qiang YAN
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 944-952.   https://doi.org/10.1631/jzus.B0820209
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The purpose of this study is to investigate the function of a novel potassium transporter gene (NrHAK1) isolated from Nicotiana rustica roots using yeast complement and real-time PCR technique. The complementary DNA (cDNA) of NrHAK1, 2 488 bp long, contains an open reading frame (ORF) of 2 334 bp encoding a protein of 777 amino acids (87.6 kDa) with 12 predicted transmembrane domains. The NrHAK1 protein shows a high sequence similarity to those of high-affinity potassium transporters in Mesembryanthemum, Phytolacca acinosa, Arabidopsis thaliana, and so on. We found that the NrHAK1 gene could complement the yeast-mutant defect in K+ uptake. Among several tissues surveyed, the expression level of NrHAK1 was most abundant in the root tip and was up-regulated when exposed to potassium starvation. Moreover, the transcript accumulation was significantly reduced by adding 5 mmol/L NH4+ to the solution. These results suggest that NrHAK1 plays an important role in potassium absorption in N. rustica.
Optimal waveband identification for estimation of leaf area index of paddy rice
Fu-min WANG, Jing-feng HUANG, Qi-fa ZHOU, Xiu-zhen WANG
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 953-963.   https://doi.org/10.1631/jzus.B0820211
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The objectives of the study were to select suitable wavebands for rice leaf area index (LAI) estimation using the data acquired over a whole growing season, and to test the efficiency of the selected wavebands by comparing them with feature positions of rice canopy spectra. In this study, the field experiment in 2002 growing season was conducted at the experimental farm of Zhejiang University, Hangzhou, China. Measurements of hyperspectral reflectance (350~2500 nm) and corresponding LAI were made for a paddy rice canopy throughout the growing season. And three methods were employed to identify the optimal wavebands for paddy rice LAI estimation: correlation coefficient-based method, vegetation index-based method, and stepwise regression method. This research selected 15 wavebands in the region of 350~2 500 nm, which appeared to be the optimal wavebands for the paddy rice LAI estimation. Of the selected wavebands, the most frequently occurring wavebands were centered around 554, 675, 723, and 1 633 nm. They were followed by 444, 524, 576, 594, 804, 849, 974, 1 074, 1 219, 1 510, and 2 194 nm. Most of them made physical sense and had their counterparts in spectral known feature positions, which indicates the promising potential of the 15 selected wavebands for the retrieval of paddy rice LAI.
Relationships between changes of kernel nutritive components and seed vigor during development stages of F1 seeds of sh2 sweet corn
Dong-dong CAO, Jin HU, Xin-xian HUANG, Xian-ju WANG, Ya-jing GUAN, Zhou-fei WANG
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 964-968.   https://doi.org/10.1631/jzus.B0820084
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The changes of kernel nutritive components and seed vigor in F1 seeds of sh2 sweet corn during seed development stage were investigated and the relationships between them were analyzed by time series regression (TSR) analysis. The results show that total soluble sugar and reducing sugar contents gradually declined, while starch and soluble protein contents increased throughout the seed development stages. Germination percentage, energy of germination, germination index and vigor index gradually increased along with seed development and reached the highest levels at 38 d after pollination (DAP). The TSR showed that, during 14 to 42 DAP, total soluble sugar content was independent of the vigor parameters determined in present experiment, while the reducing sugar content had a significant effect on seed vigor. TSR equations between seed reducing sugar and seed vigor were also developed. There were negative correlations between the seed reducing sugar content and the germination percentage, energy of germination, germination index and vigor index, respectively. It is suggested that the seed germination, energy of germination, germination index and vigor index could be predicted by the content of reducing sugar in sweet corn seeds during seed development stages.
Antiviral activities of extracts from Hong Kong seaweeds
Hui WANG, Engchoon Vincent OOI, Put O. Ang Jr.
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 969-976.   https://doi.org/10.1631/jzus.B0820154
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We extracted six Hong Kong brown seaweed species with hot water for their antiviral properties. The cytotoxicity and antiviral activity of these extracts were tested by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenlytetrezolium bromide] method, cytopathic effect reduction assay, and plaque reduction assay. The antiviral effect was further determined by flow cytometric analysis. The results showed that most of these extracts inhibited the propagation of herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) standard strains with very low cytotoxicity to the host cells. The extracts of Hydroclathrus clathratus and Lobophora variegata showed more potential anti-HSV activities than the extracts of the other four seaweeds. They also had moderate anti- respiratory syncytial virus (RSV) activities but could not inhibit influenza A virus. Hydroclathrus clathratus was further extracted by diluted acid and alkali and the antiviral effects of the extracts were also detected. The result showed that the hot water extract contained the main carbohydrate components that exhibited the antiviral activities against various strains of HSV, including the acyclovir-resistant strain. HI-3, a compound fractionated from this hot water extract, showed a dose-dependent anti-HSV activity in flow cytometric analysis and plaque reduction assay.
Analysis of volatile components in a Chinese fish sauce, Fuzhou Yulu, by gas chromatography-mass spectrometry
Yuan-fan YANG, Shen-ru CHEN, Hui NI, Xing-qian YE
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 977-981.   https://doi.org/10.1631/jzus.B0820021
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Volatile components of Fuzhou Yulu, a Chinese fish sauce, were analyzed by gas chromatography-mass spectrometry (GC-MS), and two pretreatment methods, i.e., purge and trap (P&T) GC-MS and ethyl acetate extraction followed by GC-MS, were compared. P&T-GC-MS method determined 12 components, including sulfur-containing constituents (such as dimethyl disulfide), nitrogen-containing constituents (such as pyrazine derivatives), aldehydes and ketones. Ethyl acetate extraction followed by GC-MS method detected 10 components, which were mainly volatile organic acids (such as benzenepropanoic acid) and esters. Neither of the two methods detected alcohols or trimethylamine. This study offers an important reference to determine volatile flavor components of traditional fish sauce through modern analysis methods.
Application of principal component-radial basis function neural networks (PC-RBFNN) for the detection of water-adulterated bayberry juice by near-infrared spectroscopy
Li-juan XIE, Xing-qian YE, Dong-hong LIU, Yi-bin YING
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 982-989.   https://doi.org/10.1631/jzus.B0820057
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Near-infrared (NIR) spectroscopy combined with chemometrics techniques was used to classify the pure bayberry juice and the one adulterated with 10% (w/w) and 20% (w/w) water. Principal component analysis (PCA) was applied to reduce the dimensions of spectral data, give information regarding a potential capability of separation of objects, and provide principal component (PC) scores for radial basis function neural networks (RBFNN). RBFNN was used to detect bayberry juice adulterant. Multiplicative scatter correction (MSC) and standard normal variate (SNV) transformation were used to preprocess spectra. The results demonstrate that PC-RBFNN with optimum parameters can separate pure bayberry juice samples from water-adulterated bayberry at a recognition rate of 97.62%, but cannot clearly detect water levels in the adulterated bayberry juice. We conclude that NIR technology can be successfully applied to detect water-adulterated bayberry juice.
L-Proline as an efficient and reusable promoter for the synthesis of coumarins in ionic liquid
Xiu-hong LIU, Ji-cai FAN, Yang LIU, Zhi-cai SHANG
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 990-995.   https://doi.org/10.1631/jzus.B0820079
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The effect of L-proline as a promoter on the condensation reaction of salicylaldehyde or its derivatives with ethyl acetoacetate in neutral ionic liquid [emim]BF4 was studied. All reactions were carried out under mild reaction conditions and achieved high yields. Moreover, the ionic liquid containing L-proline could be recycled and reused for several times without noticeably decreasing in productivity. The results show that the L-proline-[emim]BF4 system has a potential in contribution to the development of environmentally friendly and inexpensive processes in organic syntheses.
Representing the retinal line spread shape with mathematical functions
Yi-rong YANG, Justin WANEK, Mahnaz SHAHIDI
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2008, 9(12): 996-1002.   https://doi.org/10.1631/jzus.B0820184
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Objective: To report a mathematical function that characterizes the double-pass line spread function (LSF) of the human eye. Determining analytical functions that represent the double-pass LSF is important because it allows modeling the optical performance of the eye. Methods: Optical section retinal images, generated in normal human eyes using a modified slit-lamp biomicroscope, were analyzed to derive the double-pass LSF by plotting the intensity distribution of laser light reflected/ scattered from the vitreoretinal interface. Three mathematical functions (Lorentzian, Gaussian, exponential) were fitted to the double-pass LSF and the root mean square error (RMSE) was calculated to provide a measure of the goodness of fit. Results: The Lorentzian function provided the best representation of the double-pass LSF of normal human eyes. The full width at half maximum (FWHM) of the Lorentzian fitted curve was positively correlated with age, indicating that the double-pass LSF broadens with age. Furthermore, the goodness of fit of the Lorentzian function was significantly better in younger subjects as compared with older subjects, suggesting that the fitted function to the double-pass LSF may vary according to age. Conclusion: The results demonstrate an age-related change in the double-pass LSF width and the goodness of fit of the Lorentzian function.
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