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Isolating and Screening of Effective Decomposing Strain of Silkworm Chrysalis Protein and Study on Parts of Its Enzymatic Properties 被引量:4
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作者 孔军 涂国全 +3 位作者 邝哲师 赵祥杰 叶明强 罗国庆 《Agricultural Science & Technology》 CAS 2010年第2期8-11,共4页
[Objective] The research aimed to obtain an effectively-decomposing strain of silkworm chrysalis protein and discuss its enzymatic properties.[Method] The effectively-decomposing bacteria of protein was isolated from ... [Objective] The research aimed to obtain an effectively-decomposing strain of silkworm chrysalis protein and discuss its enzymatic properties.[Method] The effectively-decomposing bacteria of protein was isolated from the decayed silkworm chrysalis by using dilution plate and its enzymatic properties were tested after primary screening and second screening.The enzyme activity was determined and the intermediate and small molecule protein content in silkworm chrysalis was measured after solid-state fermentati... 展开更多
关键词 Decomposing strain of silkworm chrysalis isolating and screening Neutral protease Enzymatic properties
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Molecular Dynamic Simulation on the Absorbing Process of Isolating and Coating of α-olefin Drag Reducing Polymer 被引量:1
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作者 李冰 盛翔 +6 位作者 邢文国 董桂霖 刘永军 张长桥 陈祥俊 周宁宁 秦占波 《Chinese Journal of Chemical Physics》 SCIE CAS CSCD 2010年第6期630-636,745,共8页
The absorbing process in isolating and coating process of α-olefin drag reducing polymer was studied by molecular dynamic simulation method, on basis of coating theory of α-olefin drag reducing polymer particles wit... The absorbing process in isolating and coating process of α-olefin drag reducing polymer was studied by molecular dynamic simulation method, on basis of coating theory of α-olefin drag reducing polymer particles with polyurethane as coating material. The distributions of sodium laurate, sodium dodeeyl sulfate, and sodium dodeeyl benzene sulfonate on the surface of α-olefin drag reducing polymer particles were almost the same, but the bending degrees of them were obviously different. The bending degree of SLA molecules was greater than those of the other two surfactant molecules. Simulation results of absorbing and accumulating structure showed that, though hydrophobie properties of surfactant molecules were almost the same, water density around long chain sulfonate sodium was bigger than that around alkyl sulfate sodium. This property goes against useful absorbing and accumulating on the surface of α-olefin drag reducing polymer particles; simulation results of interactions of different surfactant and multiple hydroxyl compounds on surface of particles showed that, interactions of different surfaetant and one kind of multiple hydroxyl compound were similar to those of one kind of surfaetant and different multiple hydroxyl compounds. These two contrast types of interactions also exhibited the differences of absorbing distribution and closing degrees to surface of particles. The sequence of closing degrees was derived from simulation; control step of addition polymerization interaction in coating process was absorbing mass transfer process, so the more closed to surface of particle the multiple hydroxyl compounds were, the easier interactions With isoeyanate were. Simulation results represented the compatibility relationship between surfactant and multiple hydroxyl compounds. The isolating and coating processes of α-olefin drag reducing polymer were further understood on molecule and atom level through above simulation research, and based on the simulation, a referenced theoretical basis was provided for practical optimal selection and experimental preparation of α-olefin drag reducing polymer particles suspension isolation agent. 展开更多
关键词 Molecular dynamic simulation Coating process Multiple hydroxyl compound Addition polymerization Optimal selection Isolation agent
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An effective technique for isolating adult activated Schwann cells
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作者 Jifei Zhang Lianhong Jin Yuzhen Zhao 《Neural Regeneration Research》 SCIE CAS CSCD 2006年第2期103-107,共5页
BACKGROUND: Schwann cells (SCs) are neuroglial cells of peripheral nerve and play a key role in repairing peripheral nerve injury; therefore, it provides an important evidence for transplantation of SCs which are c... BACKGROUND: Schwann cells (SCs) are neuroglial cells of peripheral nerve and play a key role in repairing peripheral nerve injury; therefore, it provides an important evidence for transplantation of SCs which are characterized by active proliferation and adult high-purity in vitro after nerve injury in clinic, and also develops a new therapeutic way for nerve injury.OBJECTEVE: To investigate an effective technique for isolating adult activated Schwann cells,DESIGN: Controlled observational study.SETTING: Mudanjiang Medical College.MATERIALS: The experiment was completed at the Department of Medical Genetics of Harbin Medical University from March 2003 to April 2005. Health female Wistar rats, aged 2 months, weighting 150-160 g, were randomly divided into 3 groups with 5 in each group.METHODS: The right sciatic nerves from 15 Wistar rats were exposed and transected at the mid thigh under pentobarbital anesthesia (4 mg/kg, Lp). Seven days later, the distal segments of the predegenerated nerves were removed and used to produce adult Schwann cell cultures. The distal segment of the predegenerated nerve, 20 mm in length, was resected. The nerve was cut into pieces 1 mm in length and incubated for 3 hours under CO2 at 37 ℃ with an enzyme mixture of 0.05% collagenase/dispase. Rats were divided into 3 groups: ① Group 1: The nerve fragments were explanted in poly-L-lysine and laminin-coated dishes with BS medium from the 1st to the 6th day, On the 6^th day, the fragments were removed into a new poly-L-lysine-laminin-coated dish and the BS medium was changed to BS with 10% FBS, The nerve fragments were replaced repeatedly in the same way in new dishes on the 12^th and the 18th days. ② Group 2: For the first 3 days, the nerve fragments were fed with BS with 10% FBS. This medium was changed to BS medium on the third day. The nerve fragments were removed to another dish on day 6 and BS medium was changed to BS with 25 mI.JL FBS. Hereafter the culture method was the same as for group 1. ③ Group 3: For the first 6 days, nerve fragments were incubated in a dish not coated with poly-L-lysine and laminin, in BS medium supplemented with 8×10^7 U/L of penicillin-streptomycin. On the 6th day, the nerve fragments were removed to a poly-L-lysine-laminin-coated dish and cultured in BS with 25 mI.JL FBS, On the 12th day, the nerve fragments were explanted a second dish and fed with BS containing 100 mL/L FBS. On the 18^th day, they were explanted to a third poly-L-lysine-laminin-coated dish, SCs were obtained from all 3 dishes on the 21st day, Finally, purity and density of SCs were identified and proliferation index was calculated at the same time.MAIN OUTCOME MEASURES : Purity and density of SCs cultured with various methods in the three groups for 21 days.RESULTS : ① Isolation and proliferation of SCs: In the group 1, they increased in number after 4 days and both purity and density of cultured SCs were significantly higher than those from group 2. In the group 2, there were few fibroblasts. In the group 3, both purity and density of cultured SCs were remarkably higher than in those from groups 1 or 2. Then optimal proliferation was soon seen and the rapid expansion of SC populations suppressed the development of contaminating fibroblasts. On the 21st day, SCs proliferated to achieve maximal density and were too crowded to be counted. With Chi-square test, the data of the purity and the density were analyzed from groups 1 to 3, the result indicated X^2=430.47, P 〈 0.05. ② Characterization and proliferation rate of SCs: Immunostaining for S100 protein was evident in the cell soma and the processes of all three groups in cultures of SCs. SCs in vitro demonstrated typical bior tri-polar morphology, had oval nuclei, and stained brightly for $100. The proliferation rate of SCs was assessed with double fluorescence staining for BrdU and S100 on the 21^st day of all three groups in cultures. About 40%-50% of the total SCs in the each group showed BrdU incorporation.CONCLUSION: The method is to use predegeneration in vivo, differential speed culture supplemented with the penicillin-streptomycin in low concentration, and changing of the concentration of FBS in the BS medium from 0 to 100 mL/L. This method allows remarkable suppression of fibroblast growth and attainment of SC proliferation and purity, in a short time, from adult nerves. 展开更多
关键词 cell An effective technique for isolating adult activated Schwann cells FIGURE
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Progresses and Strategies of Gene Mapping and Isolating of Photo(thermo) sensitive Genic Male Sterile Rice
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作者 江树业 《High Technology Letters》 EI CAS 2000年第3期91-94,共4页
Considering the research on classical genetics of photoperiod(therm) sensitive genic male sterile rice, it is important to select the sterile lines and their segregating population controlled by one pair of gene in ma... Considering the research on classical genetics of photoperiod(therm) sensitive genic male sterile rice, it is important to select the sterile lines and their segregating population controlled by one pair of gene in mapping and isolating sterile genes. It is discussed the advantages, disadvantages and the reasons leading to various mapping results of chromosome location of sterile genes through gene marker, isozyme marker and DNA marker techniques. In comparison to isolation of photo(thermo) sensitive sterile genes via various plant gene cloning techniques, it was concluded that map based cloning was acceptable, but it is still difficult to locate the loci of sterile genes within 1cM. On the other hand, “sensitivity to environment”, an important characteristic of sterile lines can be fully utilized by DD PCR and (or) RDA techniques. Therefore, these two techniques were considered as the effective ways to isolate sterile genes. 展开更多
关键词 RICE PHOTOPERIOD (therm) SENSITIVE GENIC male STERILITY Gene isolating and CLONING
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A NEW METHOD OF ISOLATING KUPFFER CELLS FROM BIOPSY TISSUE OF RAT LIVER
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作者 张力健 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 1990年第3期79-81,共3页
The isolation of a high yield and purity of Kupffer cells has been reported in detail.1 This paper reports into the research about isolation Kupffer cells from biopsy tissue of liver. This method includes 5 important ... The isolation of a high yield and purity of Kupffer cells has been reported in detail.1 This paper reports into the research about isolation Kupffer cells from biopsy tissue of liver. This method includes 5 important steps: (1) take fresh liver tissue, and mince with scissors. (2) spin at low speed to wash off red blood cells. (3) digest in collagenase for suitable time. (4) isolate Kupffer cells on a percoll density gradient. (5) cell charaterization was observed by N.S.E stain and peroxidatic activity with lumino-meter measurement and phagocytosis with latex beads. 展开更多
关键词 A NEW METHOD OF isolating KUPFFER CELLS FROM BIOPSY TISSUE OF RAT LIVER EGTA
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Isolating cDNA clones from rice induced by Magnaporthe grisea using PCR based differential screening method
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作者 DONG Haitao, DONG Jixin, WU Yuliang, HE Zuhua,and Li Debao,Biotechnology Institute,ZheJiang Agri Univ, Hangzhou 310029, China 《Chinese Rice Research Newsletter》 1998年第3期1-2,共2页
The differential hybridization technique hasbeen widely used to identify genes that are dif-ferentially expressed.However,this approachhas several drawbacks.First,the screeningprocedures are rather labor-intensive and... The differential hybridization technique hasbeen widely used to identify genes that are dif-ferentially expressed.However,this approachhas several drawbacks.First,the screeningprocedures are rather labor-intensive and time-consuming.Second,the amount of phageDNAs transferred onto the two filters may notbe equivalent,which leads to an inaccurate se-lection of a positive clone.Third,isolation ofphage DNA is slow and cumbersome.Here,aPCR based differential screening method that 展开更多
关键词 cDNA PCR isolating cDNA clones from rice induced by Magnaporthe grisea using PCR based differential screening method
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A rapid approach for isolating a single fungal spore from rice blast diseased leaves 被引量:1
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作者 FEI Li-wang LU Wen-bo +8 位作者 XU Xiao-zhou YAN Feng-cheng ZHANG Li-wei LIU Jin-tao BAI Yuan-jun LI Zhen-yu ZHAO Wen-sheng YANG Jun PENG You-liang 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2019年第6期1415-1418,共4页
Single spore isolation is a fundamental approach in plant pathology and mycology to isolate and identify plant fungal pathogens from diseased samples.However,routine single spore isolation procedure is time-consuming ... Single spore isolation is a fundamental approach in plant pathology and mycology to isolate and identify plant fungal pathogens from diseased samples.However,routine single spore isolation procedure is time-consuming and has a high risk of contamination by other microorganisms.In this study,we developed a rapid approach for isolating a single spore of the fungal pathogen,Pyricularia oryzae,from rice blast diseased leaves in the paddy field with low potential of contamination.First,rice blast leaves with single lesions were selected in the paddy field,and a single lesion was cut out and pressed and dragged gently across the surface of water agar.Next,a germinated single spore with a barely visible piece of agar was cut out of water agar with a dissecting needle under a stereomicroscope at approximately 120-fold magnification.Last,the germinated single spore with water agar was transferred onto oatmeal tomato agar for growth and preservation.Based on our experience,a skilled technician or student can successfully isolate single spore from over 150 independent diseased samples with nearly no contaminations in a single working day.This approach is also suitable for isolating single spore from other fungal disease samples that produce abundant aerial conidia. 展开更多
关键词 PYRICULARIA ORYZAE SINGLE SPORE isolation rice BLAST field experiment
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Efficiency of Plating Media and Enrichment Broths for Isolating <i>Salmonella</i>Species from Human Stool Samples: A Comparison Study 被引量:2
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作者 Apichai Srijan Boonchai Wongstitwilairoong +1 位作者 Ladaporn Bodhidatta Carl Mason 《Open Journal of Medical Microbiology》 2015年第4期231-236,共6页
A comparative study was performed to evaluate best practice culture media and enrichment broths for recovering Salmonella species from human stool samples. A total of 1297 human stools were collected and processed in ... A comparative study was performed to evaluate best practice culture media and enrichment broths for recovering Salmonella species from human stool samples. A total of 1297 human stools were collected and processed in this study. Evaluation of agar media was carried out by direct plating (DP), 1096 stool samples were inoculated on Modified Semisolid Rappaport-Vassiliadis (MSRV), Xylose-Lysine-Deoxycolate (XLD), MacConkey (MAC), and Hektoen Enteric (HE) agars. Evaluation of enrichment broths were carried out by enrichment all 1297 stool samples in Selenite broth (SB), Rappaport-Vassiliadis (RV) broth, and Buffered Peptone Water (BPW), followed by plating on MSRV, MAC, and HE agars. A total of 102 Salmonella-positive stools by DP, 85.3% (87/102) were recovered utilizing MSRV while recovery from XLD, MAC, and HE agars were 34.3% (35/102), 34.3% (35/102), and 29.4% (30/102) respectively. A total 299/1297 stools samples were Salmonella-positive on at least one plating medium after enrichment procedure were 77.3% (177/299) for SB, 86.0% (197/299) and 78.6% (180/299) for RV and BPW respectively. All Salmonella isolated in this study was nontyphi Salmonella. Presently, the data suggest that the use of MSRV over MAC, HE, and XLD agars for isolation nontyphi Salmonella species from human stools is more efficacious. Additionally, use of MSRV in combination with MAC and HE agars following enrichment in RV broth enhances recovery of nontyphi Salmonella species. However, RV broth is inhibitory to typhi Salmonella, thus use of MSRV medium in combination with MAC, HE or XLD agars in direct plating following enrichment in non-selective BPW is an alternate method for recovery of both typhi and nontyphi Salmonella species contaminated in human stool samples. 展开更多
关键词 HUMAN STOOL Specimens Culture MEDIA SALMONELLA Isolation ENRICHMENT
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Isolated Pelvic Hyperthermochemotherapeutic Perfusion-An Experimental Study on Isolating Efficacy 被引量:1
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作者 张进祥 郑启昌 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2000年第3期239-241,共3页
Hyperthermochemotherapeutic perfusion model through isolated pelvic vessels was developed to evaluate the leakage of hyperthermia and drugs (such as adriamycin) from the isolated pelvic circulation to systemic circula... Hyperthermochemotherapeutic perfusion model through isolated pelvic vessels was developed to evaluate the leakage of hyperthermia and drugs (such as adriamycin) from the isolated pelvic circulation to systemic circulation and its associated side/toxic effects. The isolated pelvic circulation was perfused through a femoral artery catheter with hyperthermic (48 ℃ to 55 ℃) adriamycin solution (50 μg/ml) for 30 min. The efflux was drained through a femoral vein catheter. And the pelvic temperature was kept at the level of 43±0.5 ℃. The temperature of pelvic circulation was kept at 4 ℃ to 5 ℃ greater than the systemic/core temperature. The adriamycin concentration of pelvic efflux was 12 to 46 folds of that of systemic serum. The difference between them was very significant ( P <0.001). As the perfusion pressure was increased, which kept lower than the mean systemic artery pressure, the leakage of the adriamycin from the isolated pelvic circulation to systemic circulation was increased, but there was no significant difference between them ( P >0.05). During isolated perfusion, the systemic blood dynamics remained stable and there were no organic injuries on the important organs. It was suggested that the isolating efficacy of the modality of isolated pelvic hyperthermochemotherapeutic perfusion through vessels was rather high. The hyperthermia and drugs could be effectively limited in the isolated pelvic region with minor side effects on the systemic circulation and important organs. 展开更多
关键词 isolated perfusion chemotherapeutic hyperthmia rectal carcinoma
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Absence of postzygotic isolating mechanisms:evidence from experimental hybridization between two species of tropical sea urchins
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作者 M.Aminur RAHMAN Tsuyoshi UEHARA +2 位作者 Aziz ARSHAD Fatimah Md.YUSOFF Mariana Nor SHAMSUDIN 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2012年第10期797-810,共14页
Two reef margin species of tropical sea urchins,Echinometra sp.C(Ec) and Echinometra oblonga(Eo),occur sympatrically on Okinawa intertidal reefs in southern Japan.Hybridization between these species was examined throu... Two reef margin species of tropical sea urchins,Echinometra sp.C(Ec) and Echinometra oblonga(Eo),occur sympatrically on Okinawa intertidal reefs in southern Japan.Hybridization between these species was examined through a series of cross-fertilization experiments.At limited sperm concentrations,where conspecific crosses reached near 100% fertilization,both heterospecific crosses showed high fertilization rates(81%-85%).The compatibility of the gametes demonstrated that if gamete recognition molecules are involved in fertilization of these species,they are not strongly species-specific.We found that conspecific crosses reached peak fertilization levels much faster than did heterospecific crosses,indicating the presence of a prezygotic barrier to hybridization in the gametes.Larval survival,metamorphosis,and juvenile and adult survival of hybrid groups were nearly identical to those of their parent species.Hybrids from crosses in both directions developed normally through larval stages to sexually mature adults,indicating that neither gametic incompatibility nor hybrid inviability appeared to maintain reproductive isolation between these species.In adults,Ec×Ec crosses gave the highest live weight,followed by Eo(ova)×Ec(sperm),Ec(ova)×Eo(sperm),and Eo×Eo.Other growth performance measures(viz.,test size,Aristotle's lantern length,and gonad index) of hybrid groups and their parental siblings showed the same trends.The phenotypic color patterns of the hybrids were closer to the maternal coloration,whereas spine length,tube-foot and gonad spicule characteristics,pedicellaria valve length,and gamete sizes showed intermediate features.Adult F 1 hybrids were completely fertile and displayed high fertilization success in F 1 backcrosses,eliminating the likelihood that hybrid sterility is a postzygotic mechanism of reproductive isolation.Conversely,intensive surveys failed to find hybrid individuals in the field,suggesting the lack or rarity of natural hybridization.This strongly suggests that reproductive isolation is achieved by prezygotic isolating mechanism(s).Of these mechanisms,habitat segregation,gamete competition,differences in spawning times,gametic incompatibility or other genetic and non-genetic factors appear to be important in maintaining the integrity of these species. 展开更多
关键词 Sea urchins Echinometra HYBRIDIZATION Gamete compatibility Reproductive isolation SPECIATION
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Reproductive Isolating Mechanisms in the Bangladesh Coastal Bullfrog Hoplobatrachus litoralis and Its Congeneric Species Revealed by Crossing Experiments and Examination on Spermatogenesis of the Hybrids
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作者 Mahmudul HASAN Mohammed Mafizul ISLAM +3 位作者 Md. Mukhlesur Rahman KHAN Ratanasate WANICHANON Atsushi KURABAYASHI Masayuki NUMIDA 《Asian Herpetological Research》 SCIE CSCD 2017年第1期27-38,共12页
To elucidate reproductive isolating mechanisms in the Bangladesh coastal bullfrog Hoplobatrachus litoralis and its congeneric species, we performed crossing experiments using three species: H. litoralis, H. tigerinus... To elucidate reproductive isolating mechanisms in the Bangladesh coastal bullfrog Hoplobatrachus litoralis and its congeneric species, we performed crossing experiments using three species: H. litoralis, H. tigerinus and H. rugulosus. In addition, we conducted histological observations on spermatogenesis of the hybrids. The reciprocal hybrids between H. litoralis and H. tigerinus developed normally with somewhat lower viability at the metamorphosis stage compared with the controls. Most of the metamorphosed frogs became mature. On the other hand, almost all hybrids between female H. rugulosus and male H. litoralis or H. tigerinus died of underdevelopment at the tadpole stage, and only a few hybrids metamorphosed normally and survived to maturity. The inner structures of the testes of the control H. litoralis and H. tigerinus were completely normal, with seminiferous tubules filled with compact bundles of normal spermatozoa. Those of the reciprocal hybrids between H. litoralis and H. tigerinus were almost normal or slightly abnormal, with seminiferous tubules that contained pycnotic nuclei in addition to normal bundles of normal spermatozoa, which demonstrates slight abnormalities in spermatogenesis. In contrast, the hybrids between female H. rugulosus and male H. litoralis or H. tigerinus had no bundles of spermatozoa nor spermatids in the seminiferous tubules, which indicates entirely abnormal spermatogenesis. Meiotic chromosome figures in the reciprocal hybrids between H. litoralis and H. tigerinus showed slight abnormalities, with the occurrence of univalents and increase of rod-shaped bivalents. These results indicate that H. litoralis and H. tigerinus are not isolated from each other by hybrid inviability nor by hybrid sterility, although the hybrids showed somewhat abnormal spermatogenesis in hybrids and that H. rugulosus is isolated from both H. litoralis and H. tigerinus by incomplete hybrid inviability and complete hybrid sterility. 展开更多
关键词 Hoplobatrachus reproductive isolation interspecific hybrids SPERMATOGENESIS meiotic chromosomes
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A Simple Method for Isolating Chloroplast DNA and Mitochondria DNA from the Same Rapeseed Green Leaf Tissue
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作者 HU Zhi-yong ZHAN Gao-miao +1 位作者 WANG Han-zhong HUA Wei 《Journal of Integrative Agriculture》 SCIE CSCD 2012年第7期1212-1215,共4页
In the study, we present a fast, simple and inexpensive protocol for isolating chloroplast and mitochondrial DNA from one rapeseed leaf tissue sample. The chloroplast and mitochondria were separated from the same gree... In the study, we present a fast, simple and inexpensive protocol for isolating chloroplast and mitochondrial DNA from one rapeseed leaf tissue sample. The chloroplast and mitochondria were separated from the same green leaf tissue by differential centrifugations. The protocol is the first report that isolates plant chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) from the same sample homogenate. The organelle DNA yield is 2-10 micrograms per gram of tissue; the DNA was fully restrictable and was successfully used for sequencing. 展开更多
关键词 chloroplast DNA mitochondrial DNA organelle isolation RAPESEED
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Method for Isolating Mitochondrial DNA from Etiolated Tissue of Cabbage
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作者 Wang Shuai Wang Chao Zhang Xiao-xuan 《Journal of Northeast Agricultural University(English Edition)》 CAS 2014年第3期23-29,F0003,共8页
Isolation of high-quality mitochondrial DNA(mtDNA) is an important premise for researching molecular mechanisms in cytoplasmic male sterility of cabbage(Brassica oleracea L.var.capitata). An efficient protocol for... Isolation of high-quality mitochondrial DNA(mtDNA) is an important premise for researching molecular mechanisms in cytoplasmic male sterility of cabbage(Brassica oleracea L.var.capitata). An efficient protocol for separation and purification of mitochondria and extraction of mitochondrial DNA(mtDNA) from etiolated tissues of cabbage was developed. We took a method combined mannitol density gradient with differential centrifugation, selected appropriate rotational speed, extended DNase I treating time and changed mitochondria cracking condition. The results showed that the extracted mitochondria in this protocol had complete structure, appeared to ellipsoid and had not been contaminated with other impurities under the Jannus Green B staining. The isolated mitochondrial DNA had high purity and yield through detecting the optical density, nuclear specific primer PCR and agarose gel electrophoresis. The results indicated that mitochondrial DNA extracted by this protocol had high quality and enabled to be used in futher genetic studies. 展开更多
关键词 CABBAGE cytoplasmic male sterility(CMS) mitochondrial DNA(mtDNA) ISOLATION
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A method for isolating rat cardiomyocytes by acute enzymatic hydrolysis
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作者 Bo Liang Hui-Ling Liao 《TMR Integrative Medicine》 2019年第1期1-6,共6页
Objective: This r esea rch showed a method of isolate rat cardio myocytes though acute enzymatic hydrolysis. Method: Our method was divided into five steps: preparation, heart ext raction, perfusion, mechanical dissoc... Objective: This r esea rch showed a method of isolate rat cardio myocytes though acute enzymatic hydrolysis. Method: Our method was divided into five steps: preparation, heart ext raction, perfusion, mechanical dissociation and pu rifica tion and r ecalcifica ti on. Fir stly, make a preparation, including solutions, equipment and so on. Secondly, after anesthesia satisfactorily, open the chest of rat and take the heart out. Thirdly, transect the aorta and hang the aorta onto the Langendroff perfusion system such that the tip is just distal to the aortic valve. For th, perfuse the hea rt with diges tion solution for 17-20 min afte r Ca2+ free tyrode solution for 5 min. Finally, remove the heart from the system quickly and dissociate mechanically in the KB solution and recalcification after filter. Result: Though the microscope, good-state cardiomyocytes (clear stripes, strong sense of three-dimensional, stick well) can be observed. Conclusion: Those cells can be used to undergo medical electrophysiological experiment, including the study of iron channel. 展开更多
关键词 CARDIOMYOCYTES ACUTE isolation ACUTE ENZYMATIC hydrolysis Patch CLAMP MYOCARDIAL ELECTROPHYSIOLOGY
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phenotype-independent“label-capture-release”process for isolating viable circulating tumor cells in real-time drug susceptibility testing
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作者 Zhiqi Lao Xiaoxue Ren +8 位作者 Dehua Zhuang Lingxia Xie Yucong Zhang Wei Li Yue Chen Penghui Li Liping Tong Paul K.Chu Huaiyu Wang 《The Innovation》 2025年第5期64-73,63,共11页
Although various strategies have been proposed for enrichment of circulating tumor cells(CTCs),the clinical outcomes of CTC detection are far from satisfactory.The prevailingmethodologies for CTC detection are general... Although various strategies have been proposed for enrichment of circulating tumor cells(CTCs),the clinical outcomes of CTC detection are far from satisfactory.The prevailingmethodologies for CTC detection are generally oriented towardnaturallyoccurring targets;however,misdetection and interference are prevalent due to the diverse phenotypes and subpopulations of CTCs,which are highly heterogeneous.Here,a CTC isolation system based on the“labelcapture-release”process is demonstrated for the precise and highly efficient enrichment of CTCs fromclinical blood samples.On the basis of the abnormal glycometabolism of tumor cells,the surface of CTCs can be decorated with artificial azido groups.By utilizing bio-orthogonal plates designed with dibenzocyclooctane(DBCO)and disulfide groups,withthe aid of anti-fouling effects,CTCs labeled with azido groups can be captured through a copper-free click reaction and subsequently released via disulfide reduction.The technique has been shown to label tumor cells with the epithelial cell adhesion molecule(EpCAM)+and EpCAM~phenotypes in both adherent and suspended states.Moreover,it effectively isolates all epithelial,interstitial,and hybrid phenotypes of CTCs from clinical blood samples collected from dozens of patients across more than 10 cancer types.Compared to the clinically approved CTC detection system,our strategy demonstrates superior performance from the perspective of broad-spectrum and accurate recognition of heterogeneous CTCs.More importantly,most of the captured CTCs can be released with the retention of living activity,making this technique well suited for downstream applications such as drug susceptibility tests involving viable CTCs. 展开更多
关键词 circulating tumor cells ctcs bio orthogonal click reaction label capture release real time drug susceptibility testing isolation system disulfide reduction phenotype independent circulating tumor cells
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Utilizing engineered extracellular vesicles as delivery vectors in the management of ischemic stroke:a special outlook on mitochondrial delivery 被引量:1
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作者 Jiali Chen Yiyang Li +7 位作者 Xingping Quan Jinfen Chen Yan Han Li Yang Manfei Zhou Greta Seng Peng Mok Ruibing Wang Yonghua Zhao 《Neural Regeneration Research》 SCIE CAS 2025年第8期2181-2198,共18页
Ischemic stroke is a secondary cause of mortality worldwide,imposing considerable medical and economic burdens on society.Extracellular vesicles,serving as natural nanocarriers for drug delivery,exhibit excellent bioc... Ischemic stroke is a secondary cause of mortality worldwide,imposing considerable medical and economic burdens on society.Extracellular vesicles,serving as natural nanocarriers for drug delivery,exhibit excellent biocompatibility in vivo and have significant advantages in the management of ischemic stroke.However,the uncertain distribution and rapid clearance of extracellular vesicles impede their delivery efficiency.By utilizing membrane decoration or by encapsulating therapeutic cargo within extracellular vesicles,their delivery efficacy may be greatly improved.Furthermore,previous studies have indicated that microvesicles,a subset of large-sized extracellular vesicles,can transport mitochondria to neighboring cells,thereby aiding in the restoration of mitochondrial function post-ischemic stroke.Small extracellular vesicles have also demonstrated the capability to transfer mitochondrial components,such as proteins or deoxyribonucleic acid,or their sub-components,for extracellular vesicle-based ischemic stroke therapy.In this review,we undertake a comparative analysis of the isolation techniques employed for extracellular vesicles and present an overview of the current dominant extracellular vesicle modification methodologies.Given the complex facets of treating ischemic stroke,we also delineate various extracellular vesicle modification approaches which are suited to different facets of the treatment process.Moreover,given the burgeoning interest in mitochondrial delivery,we delved into the feasibility and existing research findings on the transportation of mitochondrial fractions or intact mitochondria through small extracellular vesicles and microvesicles to offer a fresh perspective on ischemic stroke therapy. 展开更多
关键词 delivery engineering extracellular vesicles identification ischemic stroke isolation MITOCHONDRIA targeting strategy therapeutic effects
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Dynamic response of concrete pavement structure with asphalt isolating layer under moving loads 被引量:9
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作者 Jianmin Wu Jiaping Liang Sanjeev Adhikari 《Journal of Traffic and Transportation Engineering(English Edition)》 2014年第6期439-447,共9页
A three-dimensional finite element model (3D FEM) is built using ABAQUS to analyze the dynamic response of a concrete pavement structure with an asphalt isolating layer under moving loads. The 3D model is prepared a... A three-dimensional finite element model (3D FEM) is built using ABAQUS to analyze the dynamic response of a concrete pavement structure with an asphalt isolating layer under moving loads. The 3D model is prepared and validated in the state of no asphalt isolating layer. Stress and deflection at the critical load position are calculated by changing thickness, modulus of isolating layer and the combination between the isolating layer and concrete slab. Analysis result shows that the stress and deflection of the concrete slab increase with the increase of thickness. The stress and deflection of the concrete slab decrease with the increase of combination between the isolating layer and concrete slab. The influence of changing the isolating layer modulus to the stress and deflection of the concrete slab is not significant. From the results, asphalt isolating layer design is suggested in concrete pavement. 展开更多
关键词 concrete pavement asphalt isolating layer moving loads three-dimensional finite element
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Rapid and efficient method for isolating plant high molecular weight(HMW)DNA with high purity 被引量:4
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作者 Fang Qiu Jianmin Fu +2 位作者 Weiwu Xie Chuanyou Li Bin Wang 《Chinese Science Bulletin》 SCIE EI CAS 1999年第4期378-380,共3页
PREPARATION of HMW DNA (Megabase-size) is the basis for construction of genomic library with large DNA inserts such as bacterial artificial chromosome (BAC) and yeast artificial chromosome (YAC), and for long-range ph... PREPARATION of HMW DNA (Megabase-size) is the basis for construction of genomic library with large DNA inserts such as bacterial artificial chromosome (BAC) and yeast artificial chromosome (YAC), and for long-range physical mapping. It can also be used for the macro-study of repeat sequences. Since HMW DNA during preparation is inclined to be sheared physically and digested by internal nucleases, it is very difficult to prepare the HMW DNA. Initially, plant HMW DNA was prepared by embedding protoplasts in the low melting-point (LMP) agarose; however, it had several disadvantages: (ⅰ) Culture of protoplasts was time-consuming, costly and tedious. ( ⅱ ) It was only used successfully for limited 展开更多
关键词 DNA Rapid and efficient method for isolating plant high molecular weight HIGH
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Identification and distribution patterns of the ultra-deep small-scale strike-slip faults based on convolutional neural network in Tarim Basin,NW China 被引量:1
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作者 Hao Li Jun Han +4 位作者 Cheng Huang Lian-Bo Zeng Bo Lin Ying-Tao Yao Yi-Chen Song 《Petroleum Science》 2025年第8期3152-3167,共16页
The isolated fracture-vug systems controlled by small-scale strike-slip faults within ultra-deep carbonate rocks of the Tarim Basin exhibit significant exploration potential.The study employs a novel training set inco... The isolated fracture-vug systems controlled by small-scale strike-slip faults within ultra-deep carbonate rocks of the Tarim Basin exhibit significant exploration potential.The study employs a novel training set incorporating innovative fault labels to train a U-Net-structured CNN model,enabling effective identification of small-scale strike-slip faults through seismic data interpretation.Based on the CNN faults,we analyze the distribution patterns of small-scale strike-slip faults.The small-scale strike-slip faults can be categorized into NNW-trending and NE-trending groups with strike lengths ranging 200–5000 m.The development intensity of small-scale strike-slip faults in the Lower Yingshan Member notably exceeds that in the Upper Member.The Lower and Upper Yingshan members are two distinct mechanical layers with contrasting brittleness characteristics,separated by a low-brittleness layer.The superior brittleness of the Lower Yingshan Member enhances the development intensity of small-scale strike-slip faults compared to the upper member,while the low-brittleness layer exerts restrictive effects on vertical fault propagation.Fracture-vug systems formed by interactions of two or more small-scale strike-slip faults demonstrate larger sizes than those controlled by individual faults.All fracture-vug system sizes show positive correlations with the vertical extents of associated small-scale strike-slip faults,particularly intersection and approaching fracture-vug systems exhibit accelerated size increases proportional to the vertical extents. 展开更多
关键词 Small-scale strike-slip faults Convolutional neural network Fault label Isolated fracture-vug system Distribution patterns
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Vibration safety assessment and parameter analysis of buried oil pipelines based on vibration isolation holes under strong surface impact 被引量:1
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作者 Wang Guobo Mei Hua +4 位作者 Wang Jianning He Wei Yin Yao Zhai Yuxin Zuo Pengfei 《Earthquake Engineering and Engineering Vibration》 2025年第1期69-82,共14页
Strong surface impact will produce strong vibration,which will pose a threat to the safety of nearby buried pipelines and other important lifeline projects.Based on the verified numerical method,a comprehensive numeri... Strong surface impact will produce strong vibration,which will pose a threat to the safety of nearby buried pipelines and other important lifeline projects.Based on the verified numerical method,a comprehensive numerical parameter analysis is conducted on the key influencing factors of the vibration isolation hole(VIH),which include hole diameter,hole net spacing,hole depth,hole number,hole arrangement,and soil parameters.The results indicate that a smaller ratio of net spacing to hole diameter,the deeper the hole,the multi-row hole,the hole adoption of staggered arrangements,and better site soil conditions can enhance the efficiency of the VIH barrier.The average maximum vibration reduction efficiency within the vibration isolation area can reach 42.2%.The vibration safety of adjacent oil pipelines during a dynamic compaction projection was evaluated according to existing standards,and the measurement of the VIH was recommended to reduce excessive vibration.The single-row vibration isolation scheme and three-row staggered arrangement with the same hole parameters are suggested according to different cases.The research findings can serve as a reference for the vibration safety analysis,assessment,and control of adjacent underground facilities under the influence of strong surface impact loads. 展开更多
关键词 vibration isolation hole buried oil pipeline strong surface impact vibration velocity vibration safety assessment
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