16 results on '"Yang, Jianping"'
Search Results
2. Feature extraction and discrimination of fabric frictional sounds.
- Author
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YANG Jianping, MENG Xu, XU Rongrong, and TAO Chen
- Subjects
FEATURE extraction ,SOUNDS ,TEXTILES ,SOUND ,FRICTION - Abstract
To discriminate different fabric frictional sound, a mechanism based on Haar features was established in this study, and an efficient approach was conducted to identify fabric friction sounds by discriminating against the performances of different fiber materials manifested in the mechanism. 4 categories of fabrics were put through a specialized apparatus to collect frictional sound signals. The Haar features on every scale and position of the acquired signal were extracted so as to establish a feature space. For each point in the feature space, a discriminator was built to approve all positive samples of a certain category and deny negative samples as many as possible. All negatives were filter out due to the probability of a single negative sample being allowed by a large number of discriminators is very small. The progressive selection was performed on the discriminators to form a queue so as to bring out a much reduced version of the unordered discriminators with the same discriminability. Results have indicated that the discrimination queue suggested by this study can substantially reduce the number of discriminators involved and meanwhile keep the discriminability of fabric friction sounds invariable. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
3. [Recognition of Low Arousal Level Electroencephalogram in the Vigilance Based on Wavelet Packet Rhythm and Support Vector Machine].
- Author
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Yang J, Zhang D, Luo W, and Xiao X
- Subjects
- Humans, Signal Processing, Computer-Assisted, Arousal, Brain physiology, Electroencephalography, Support Vector Machine, Wavelet Analysis
- Abstract
Poor and monotonous work could easily lead to a decrease of arousal level of the monitoring work personnel. In order to improve the performance of monitoring work, low arousal level needs to be recognized and awakened. We proposed a recognition method of low arousal by the electroencephalogram (EEG) as the object of study to recognize the low arousal level in the vigilance. We used wavelet packet transform to decompose the EEG signal so the EEG rhythms of each component were obtained, and then we calculated the parameters of relative energy and energy ratio of high-low frequency, and constructed the feature vector to monitor low arousal state in the operation. We finally used support vector machine (SVM) to recognize the low arousal state in the simulate operation. The experimental results showed that the method introduced in this article could well distinguish low arousal level from arousal level in the vigilance and it could also get a high recognition rate. Have been compared with other analysis methods, the present method could more effectively recognize low arousal level and provide better technical support for wake-up mechanism of low arousal state.
- Published
- 2016
4. [Status analysis on child neglect of rural pupils aged 6-11 year-old in China].
- Author
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Pan J, Yang W, Jiao F, Zhang Y, Zhang H, Chen J, Xi W, Peng Y, Gu G, Zhong Z, Qin R, Li J, Chu Y, Yang J, Ma L, Wang W, Wu W, Yang W, Ying Y, Gao Y, Teng H, and Liu T
- Subjects
- Child, China, Female, Humans, Male, Physical Examination, Pupil, Risk Factors, Schools, Surveys and Questionnaires, Child Abuse, Rural Population
- Abstract
Objective: To analyze pupils' neglect status aged 6-11 years in China's rural., Methods: According to the principle of multi-stage stratified cluster sampling, to extract 7,943 pupils aged 6-11 years for the survey from 56 primary schools in 28 counties in nine provinces, from December 2012 to March 2013. Proceed questionnaire survey by the scale from "Neglect Evaluation Norms for pupils aged 6-8 years in Rural Areas of China" and "Neglect Evaluation Norms for pupils aged 9-11 years in Rural Areas of China". And analyze neglect rates and neglect degrees of the different grades, gender, family types and different levels of neglect., Results: The total neglect rate of China's rural pupils aged 6-8 and 9-11 years was 40.2% (1,258/3,130) and 42.5% (1,498/3,526) respectively, which differences had no statistical significant (χ2=3.59, P=0.058); the total neglect degree was 46.04±8.87 and 44.78±10.43 respectively, which differences had statistical significant (t=5.01, P<0.001); the differences of neglect rates and neglect degrees between male (41.4% (657/1,587) and 46.28±8.76) and female (39.0% (601/1,543) and 45.78±8.97) of aged 6-8 were all no statistical significant (χ2=1.87, P=0.171; t=1.49, P=0.136); the neglect rates and neglect degrees of male (46.3% (816/1,763) and 45.53±10.11) were higher than female (38.7% (682/1,763) and 44.06±10.69) in the group of aged 9-11, which differences were all statistical significant (χ2=20.84, t=3.97, P<0.001); the male neglect rate in the group of aged 6-8 in social neglect (11.7% (198/1,691)) and the neglect degree in educational neglect (48.09±9.70) were higher than female (9.4% (155/1,648) and 47.37±9.89), which differences were all statistical significance (χ2=14.55, P<0.001, t=2.22, P=0.026), the male neglect rate in the group of aged 9-11 in physical neglect (20.4% (398/1,954)) was higher than female (16.7% (326/1 957)), which differences had statistical significance (χ2=8.92, P=0.003)., Conclusion: The neglect status of Chinese pupils aged 6-11 years in rural was serious, and we should find out risk factors and provide efficient prevention measures.
- Published
- 2015
5. [Intracerelar ventricular injection of the antisense oligonucleotides of ERK5 inhibits neuropathic pain].
- Author
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Chen Q and Yang J
- Subjects
- Animals, Cyclic AMP Response Element-Binding Protein, Hot Temperature, Hyperalgesia, Injections, Oligonucleotides, Antisense, Rats, Signal Transduction, Spinal Cord, Mitogen-Activated Protein Kinase 7 antagonists & inhibitors, Neuralgia
- Abstract
Objective: To explore the effects of extracellular signal-regulated protein kinase 5 signaling pathway in chronic constriction injury (CCI)-induced neuropathic pain above spinal cord level., Methods: The CCI model of neuropathic pain was established. Intracerebroventricular injection of antisense oligonucleotides and immunohistochemical labeling technique were employed. And the heat and mechanical hyperalgesia responses were used to examine the affect of ERK5 in CCI-induced neuropathic pain and the expression of p-CREB (cAMP response-element binding protein) in spinal cord., Results: The mechanical paw withdrawal reflex threshold and thermal reflex latency became significantly lower in CCI-induced mice. Compared with saline group, mechanical and thermal hyperalgesia significantly decreased and the expression of p-CREB protein decreased at spinal cord in oligonucleotide group., Conclusion: The ERK5 spinal cord neurons are involved in the regulation of CCI-induced neuropathic pain in mice. And this effect is achieved by adjusting the CREB-dependent gene expression.
- Published
- 2014
6. [Development of Neglect Evaluation Scale for primary school students aged 6-11 years old in rural areas of China].
- Author
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Pan J, Yang W, Chen J, Xi W, Zhang H, Peng Y, Gu G, Zhang Y, Zhong Z, Jiao F, Qin R, Li J, Zhu Y, Yang J, Wang W, Ma L, Teng H, Gao Y, Ying Y, Wu W, Yang W, and Liu T
- Subjects
- Child, China, Factor Analysis, Statistical, Humans, Reproducibility of Results, Schools, Child Abuse, Rural Population, Students, Surveys and Questionnaires
- Abstract
Objective: To develop a standard Neglect Evaluation Scale which is suitable for rural primary school students in China., Methods: According to the principle of multi-stage stratified cluster sampling, a field investigation was conducted among primary school students in 28 counties from 7 provinces and 2 municipalities. The questionnaires were self-designed, and determined by multiple rounds of pilots and revisions. Among 5 862 students, 2 792 (6-8 years' old) and 3 070 (9-11 years' old) were investigated by using two kinds of Neglect Evaluation questionnaires, respectively. After project analysis, factor analysis, reliability and validity analysis, the reliability and stability of the scale were tested. Then percentile method was used to determine the evaluation standard to develop and finalize the formal scale., Results: The numbers of the remaining items for 6-8 year-old group and 9-11 year-old group is 69 and 58, respectively, both of which contain six levels of neglect, including body, emotion, health care, education, security, and society. 4 times of factor analysis were conducted in both of the two groups. The factor loadings in these two groups were 0.290-0.700 and 0.276-0.729 respectively. Reliability test results showed that the two kinds of scales' Cronbach alpha coefficient were 0.924 and 0.929 respectively, split-half reliability were 0.891 and 0.904 respectively, the retest reliability were 0.559 and 0.892 respectively, the differences were statistically significant (P < 0.05). The retest reliability among 6-8 year-old group in medical neglect level had no statistical significance, but was close to the cut-off point (P = 0.054). The test results of external validity indicated that both of the two scales could reflect the neglect status of the subjects (P < 0.05), but the subjective and objective evaluation towards neglect were not consistent. The cut-off points for judging whether the children were neglected or not among 6-8 year-old and 9-11 year-old groups were 159 and 137 respectively; and the adjusted values were 160 and 135 respectively., Conclusion: The two norm scales developed by this study showed good discriminability, reliability, validity, and stability. The norms developed on the basis of the scales was suitable for the situation of rural students in primary schools in China.
- Published
- 2014
7. [Gravity frequency and its monitoring application of EEG spectrum in the vigilance operation].
- Author
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Yang J, Zhang D, Lu J, and Shuai X
- Subjects
- Algorithms, Humans, Brain physiology, Electroencephalography
- Abstract
It is an important means to study the electrical activity of the brain's nerve cells by exploring physiological information of the EEGs from the frequency domain. The gravity frequency is one of the global parameters with using this method. We used the multitaper spectrum method (MTM) spectrum estimation method of good performance to calculate the EEG spectrum and its gravity frequency of subjects under vigilance and vigilance decrement state. The results showed that the gravity frequency of vigilance state was higher than that of vigilance decrement state, the gravity frequency became smaller along with the vigilance decrement, and the location of the gravity frequency shifted to the left in the spectrum. Finally, the monitoring curve of the gravity frequency was acquired by designing an algorithm, and it was used to online monitoring vigilance operators.
- Published
- 2014
8. [Bedside ultrasound rapid diagnosis on pleural effusion and pneumothorax of H7N9 infected acute respiratory distress syndrome patients].
- Author
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Zhang Y, Yang J, and Chen J
- Subjects
- Humans, Influenza A Virus, H7N9 Subtype, Influenza, Human complications, Male, Middle Aged, Pleural Effusion etiology, Pneumothorax etiology, Respiratory Distress Syndrome complications, Respiratory Distress Syndrome virology, Ultrasonography, Pleural Effusion diagnostic imaging, Pleural Effusion therapy, Pneumothorax diagnostic imaging, Pneumothorax therapy
- Published
- 2014
- Full Text
- View/download PDF
9. [Profile of P66SHC expression and histone modifications in replicative cell senescence and oxidative-stress induced premature senescence].
- Author
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Xu W, Zhuang Z, Yang J, Yang L, Xu Y, and Zhang W
- Subjects
- Cell Proliferation, Fibroblasts cytology, Gene Expression Profiling, Histones metabolism, Humans, Hydrogen Peroxide, Lung cytology, Shc Signaling Adaptor Proteins genetics, Src Homology 2 Domain-Containing, Transforming Protein 1, Aging, Premature chemically induced, Aging, Premature genetics, Aging, Premature physiopathology, Cellular Senescence genetics, Histones genetics, Oxidative Stress physiology, Shc Signaling Adaptor Proteins metabolism
- Abstract
Objective: To study the profile of P66SHc expression and histone modifications in replicatively senescenct cells and oxidative-stress inducing premature senescenct cells., Methods: HPF cells were continuously cultured and subcultured in vitro to build replicative cellular model. HPF cells were treated with 200 pmol/L H2 O2 four times to build oxidative-stress inducing premature senescenct model. Comparative Q-PCR was utilized to investigate target gene (P66SHC, EP300, HDAC1) expressions respectively in H2O2 treated groups and normal cell groups. Then CHIP-QPCR was conducted to analyze histone modifications of P66SHC between young cells and aging cells., Results: P66SHC expression was positive correlation with H2O2 doses and population doubling level (PDL) (R = 0.909, P = 0.000; R = 0.743, P = 0.006), while EP300 was negative correlation with H2O2 (R = - 0.922, P = 0.000) and both EP300 and HDAC1 were negative with PDL (R = -0.709, P = 0.010, R = -0.599, P = 0.040). H3 histone modifications were declined in P66SHc gene regulating region. H3-Ac, H3K9-Ac and H3K4-tri-Me were dominant in the upstream region of transcriptional site (-3.0 kb) and alternative promotor (+3.8 kb)., Conclusion: P66SHC, EP300 and HDAC1 probably play a role in cellular replicative senescence and oxidative-stress inducing premature senescence. Besides, histone modification could regulate P66SHC gene expression.
- Published
- 2013
10. [An analytic method of wavelet energy value to evaluate the contraction intensity of uterus].
- Author
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Yang J and Xiao K
- Subjects
- Artifacts, Female, Humans, Pregnancy, Wavelet Analysis, Algorithms, Signal Processing, Computer-Assisted, Uterine Contraction physiology, Uterine Monitoring
- Abstract
The data of uterine contraction pressure is the information source for extracting uterine contractions status. Because there is a variety of interference existing in contraction pressure data, commonly used methods such as uterine contraction intensity integration method can not obtain decent evaluation results. We used the bior 2.4 biorthogonal wavelet to decompose and reconstruct the pressure data in order to obtain the best denoising effect. Combining with the denoised results, we proposed an algorithm of the wavelet energy value. Based on the algorithm, we calculated the curve of wavelet energy value. It was proved that using the curve of wavelet energy value can better identify contractions waveform and evaluation contractions intensity.
- Published
- 2012
11. [Effect of hydroquinone on the expression of ubiquitin-conjugating enzyme Rad6B in human L-02 hepatic cells].
- Author
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Hu G, Zhuang Z, Yang J, and Huang H
- Subjects
- Cell Line, Hepatocytes cytology, Humans, Mutagens toxicity, RNA, Messenger genetics, RNA, Messenger metabolism, Ubiquitin-Conjugating Enzymes genetics, Up-Regulation drug effects, Hepatocytes enzymology, Hydroquinones toxicity, Ubiquitin-Conjugating Enzymes metabolism
- Abstract
Objective: To study the effects of hydroquinone (HQ) on expression of ubiquitin-conjugating enzyme Rad6B in human L-02 hepatic cells., Methods: After L-02 hepatic cells were at the concentrations of 0, 5, 10, 20, 40, 80, and 160 micromol/L HQ for 24h, and the levels of Rad6B mRNA and protein expressions in L-02 hepatic cells were detected by real-time fluorescent quantitative polymerase chain reaction (QPCR) technique and Western blot method respectively., Results: It was found that HQ in the range of 0-160 micromol/L could induce increases in the expressions of Rad6B mRNA and protein, which was in a dose-dependent manner. The levels of Rad6B mRNA and protein expressions in the treated group of 5, 10, 20, 40, 80, and 160 micromol/L were higher than those in the control (P < 0.01), levels of Rad6B mRNA and protein expressions reached the maximun when L-02 hepatic cells were treated by HQ at the concentrations of 160 micromol/L, and the relative quantity values were 4.35 and 0.85, respectively., Conclusion: HQ could regulate up the expression of Rad6B in L-02 hepatic cells.
- Published
- 2009
12. [Study on the replicative and premature senescence induced by hydrogen peroxide of human embryonic lung fibroblasts].
- Author
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Zhang W, Ji W, Yang L, and Yang J
- Subjects
- Cell Proliferation drug effects, Cells, Cultured, Embryo, Mammalian, Humans, Proliferating Cell Nuclear Antigen genetics, Proliferating Cell Nuclear Antigen metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, beta-Galactosidase metabolism, Cellular Senescence drug effects, Fibroblasts cytology, Hydrogen Peroxide pharmacology, Lung cytology
- Abstract
Objective: To study both cellular replicative and prematuce senescence induced by hydrogen peroxide (H2O2) of human embryonic lung fibroblasts (HEFs)., Methods: HEFs were cultured conventionally and treated with H2O2. Biomarkers of senescence were assessed, including cell morphology, growth curve, life span, distribution of cell cycle and senescence associated beta-galactosidase (SA-beta-gal), and the expression of proliferating cell nuclear antigen (PCNA)., Results: HEFs entered replicative senescence status at 52 population doubling levels (PDL) and 400 micromol/L H2O2 induced premature senescence of HEFs in a short period. Senescent cells were characterized as enlarged, flattened and reduced saturation density, retarded in the G1 phase with increased expression of SA-beta-gal. The mRNA and protein level of PCNA decreased gradually during cellular senescence., Conclusion: Cellular premature senescence could be induced by 400 micromol/L H2O2. Premature senescence induced by H2O2 could share the morphology characteristics and proliferative capacity similar to those of replicative one.
- Published
- 2009
13. [An experimental study on treatment of steroid-associated femoral head necrosis with simvastatin and BMSCs transplantation].
- Author
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Yang J, Wang L, Xu Y, Wang J, and Wang Y
- Subjects
- Animals, Cells, Cultured, Disease Models, Animal, Femur Head Necrosis chemically induced, Femur Head Necrosis pathology, Male, Methylprednisolone toxicity, Rabbits, Random Allocation, Simvastatin administration & dosage, Treatment Outcome, Bone Marrow Transplantation methods, Femur Head Necrosis drug therapy, Femur Head Necrosis surgery, Simvastatin therapeutic use, Stem Cell Transplantation methods
- Abstract
Objective: To approach the possibility of combination of simvastatin and BMSCs transplantation for steroid-associated osteonecrosis of femoral head., Methods: The BMSCs harvested from 24 rabbits were prepared for cell suspension at a concentration of 1 x 107/mL, and combined with gelatin sponge. Seventy New Zealand white rabbits received one intravenous injection of lipopolysaccharide (10 microg/kg). After 24 hours, three injections of 20 mg/kg of methylprednisolone were given intramuscularly at a time interval of 24 hours. Forty-eight rabbits diagnosed as having femoral head necrosis by MRI were divided into 4 groups randomly, group A: no treatment; group B: only decompression; group C: decompression and BMSCs transplantation; and group D: simvastatin drench (10 mg/kg.d) decompression and BMSCs transplantation. The general information of animals were recorded; after 4 and 8 weeks of operation, 6 rabbits of each group were chosen randomly to do MRI scan, and femoral heads were harvested to do histopathology and scanning electron microscope examination., Results: After 8 weeks, rabbits became more active than before treatment, and walking way became normal gradually in groups C and D. Four weeks after operation, the MRI low signal region of all groups had no obvious changes, but 8 weeks later, the necrosis signal region of group A magnified while it reduced obviously in group D. Histopathological observation: 4 weeks after operation, diffuse presence of empty lacunae and pyknotic nuclei of osteocytes were found in the trabeculae, and few newborn micrangium could been seen in group A; lots of empty lacunae and a small quantity of newborn micrangium could been found in group B; and large amounts of osteoblats and newborn micrangium were found around the necrosis regions in groups C and D. The positive ratio of empty lacunae and microvessel density in group D were 19.30 +/- 1.52 and 7.08 +/- 1.09, showing significant difference compared with other groups (P < 0.05). After 8 weeks of treatment, the bone trabecula collapsed in many regions in group A; there was fibra callus formation along the decompression channel in group B; few empty lacunae was in the bone trabecular, but the shape of marrow cavity was not normal in group C; and it showed almost normal appearance in group D. The positive ratio of empty lacunae and microvessel density in group D were 11.31 +/- 1.28 and 12.37 +/- 1.32, showing significant differences compared with other groups (P < 0.05), meanwhile, showing significant difference compared with that of 4 weeks after operation(P < 0.05). Scanning electron microscope: 8 weeks after operation, the bone trabecula collapsed in many regions, and few osteoblasts could be found on the surface, a great quantity of fat cells cumulated in the bone marrow in group A; cracked bone trabecula could be found occasionally in group B; the density of bone trabecula was lower than the normal in group C; and the shape of the marrow cavity and the density of bone trabecula were similar to the normal in group D., Conclusion: Simvastatin can promote the differentiation of osteocyte and vascular endothelial cell from MSCs, the combination of simvastatin and marrow stem cells transplantation for the treatment of steroid-associated osteonecrosis of femoral head have good application prospects.
- Published
- 2008
14. [An experimental osteonecrosis of femoral head induced by a combination of a single low-dose lipopolysaccharide and methylprednisone].
- Author
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Yang J, Wang L, Xu Y, Fan K, and Wang J
- Subjects
- Adipocytes pathology, Animals, Bone Marrow Cells drug effects, Bone Marrow Cells pathology, Disease Models, Animal, Dose-Response Relationship, Drug, Femur Head diagnostic imaging, Femur Head Necrosis blood, Femur Head Necrosis chemically induced, Hormones blood, Lipoproteins, HDL blood, Lipoproteins, LDL blood, Magnetic Resonance Imaging, Male, Plasminogen Activator Inhibitor 1 blood, Rabbits, Radiography, Random Allocation, Tissue Plasminogen Activator blood, Femur Head pathology, Femur Head Necrosis pathology, Lipopolysaccharides toxicity, Methylprednisolone toxicity
- Abstract
Objective: To investigate both incidence and mechanism attributing to steroid-associated osteonecrosis of femoral head (ONFH) using an experimental protocol with a single low-dose lipopolysaccharide (LPS) injection and subsequently three injections of high-dose methylprednisolone (MPS)., Methods: Twenty-five New Zealand white rabbits with body weight of (3.0 +/- 0.3) kg were divided randomly into 2 groups. In treatment group, 19 rabbits received one intravenous injection of LPS (10 microg/kg); 24 hours later, three injections of 20 mg/kg of MPS were given intramuscularly at an interval of 24 hours. Additional 6 rabbits which received normal saline injection at the same time point were used as controls (control group). The blood samples were collected for hematological examinations before and after LPS injection, MRI was performed on bilateral hip six weeks after last MPS injection, meanwhile, bone marrow was aspirated from femoral head region to evaluate stem cell's activity. Bilateral femoral heads were harvested to make histopathology examination., Results: All animals survived throughout the experiment period except one death on the second day after LPS injection. In the histopathological examination for the femoral head, ONFH+ was observed in 16 rabbits (88.9%), and the lesions were mainly in the metaphysis. In ONFH+ rabbits, micro vessels fibrous thrombosis and extravascular marrow fat cell size increasing were found around necrotic bone; The femoral heads of control group had no changes. MRI accurate ratio was 93.8% (15/16). Compared to baseline, a significant decrease in ratio of tissue plasminogen activator/plasminogen activator inhibitor 1 and activated partial thromboplatin time, and a significant increase in ratio of low-density lipoprotein/high-density lipoprotein were only found in ONFH+ rabbits (P < 0.05). Meanwhile there was a significant decrease in the number of CFU-F (8.50 +/-9.63) compared with the control (70.17 +/- 7.78, P < 0.05)., Conclusion: A single low-dose LPS injection and subsequent three injections of high-dose MPS is effective on building steroid-associated ONFH model, coagulation and lipometabolism abnormality, activity degeneration of stem cell may be the key factors of ONFH.
- Published
- 2008
15. [Construction of short hairpin RNA vector of inhibiting poly ADP-ribose polymerase activity].
- Author
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Sha Y, Zhuang ZX, He Y, Hu DL, Hu G, Yang J, and Tu X
- Subjects
- Humans, Poly (ADP-Ribose) Polymerase-1, RNA Interference, Genetic Vectors genetics, Poly(ADP-ribose) Polymerases genetics, Poly(ADP-ribose) Polymerases metabolism, RNA, Small Interfering genetics
- Abstract
Objective: To construct expressing vector of short hairpin RNA (shRNA) in order to inhibit human PARP1 activity., Methods: 2 pairs of 64 base oligos for hairpin RNA expression which targeted PARP1 gene were chemically synthesized and annealed then ligased with pSIREN-RetroQ vector with BamH II and EcoR I . Cut by EcoR t and Bgl II, shRNA and its upstream U6, which have 330 bp, were inserted into the same treated pEGFP-C1 vecter to construct GFP expression plasmids that inhibited hPARP1 protein shRNA plasmid (pEGFP-C1P). Oligos with a scrambled sequence were used as a negative control., Results: Recombinant pEGFP-C1P1, pEGFP-C1P2 and pEGFP-C1N vectors was identified by digestion with EcoR I and Bgl II and confirmed by sequencing analysis with U6 primer. The results demonstrated that the 330 bp had been inserted into the expected site. Furthermore, the insertion sequence was exactly correct., Conclusion: pEGFP-C1-shRNA system has been constructed successfully. This will facilitate the study of PARP1's DNA repairing function.
- Published
- 2006
16. [Construction of "pEGFP-C1-pU6-dsRNA" recombinant for human DNA polymerase beta RNA interference].
- Author
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Hu DL, Zhuang ZX, He Y, Ji WD, Tang D, Yuan J, Fang D, Sha Y, Yang J, Tu X, and Hu G
- Subjects
- Cloning, Molecular, Environmental Pollutants adverse effects, Genetic Vectors genetics, Green Fluorescent Proteins genetics, Humans, RNA Interference, RNA, Double-Stranded biosynthesis, Recombinant Proteins biosynthesis, Recombinant Proteins genetics, DNA Polymerase beta genetics, DNA Repair genetics, RNA, Double-Stranded genetics, RNA, Small Interfering genetics, RNA, Small Nuclear genetics
- Abstract
Objective: To clone the "pEGFP-C1-pU6-dsRNA" recombinant for human DNA polymerase beta RNA interference, to provide research tool for the study on the function of DNA polymerase beta in repairing of human DNA damaged by environmental chemical pollutants (ECPs)., Methods: According to the gene sequence of polymerase beta cDNA published in Genbank, double strand RNA(dsRNA) sequence which was used in RNA interference was designed by dsRNA oligonucleotide designer and synthesized by chemical methods. DNA recombination technology was used to insert the up related dsRNA sequence into the vector of pSIREN-RetroQ, and then the "pSIREN-RetroQ-dsRNA" recombinant was obtained. After E. coli DH5alpha was transformed with the "pSIREN-RetroQ-dsRNA" recombinant and screened with ampicillin for positive clones, plasmid was extracted and digested by EcoR I and Bgl II , the fragment of"pU6-dsRNA"was purified. And then the "pU6-dsRNA"fragment was cloned into the vector of pEGFP-C1 by recombination technology, the recombinant of "pEGFP-C1-pU6-dsRNA" was obtained and identified by restriction endonuclease analysis and sequencing., Results: The "pEGFP-C1-pU6-dsRNA" recombinant lied in the predicted band, and the sequence of insert was identical to the designed target fragment., Conclusion: The "pEGFP-C1-pU6-dsRNA" recombinant was successfully cloned for human DNA polymerase beta RNA interference, it was an important research tool for the further study.
- Published
- 2006
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