1.Cloning, Expression and Property Analysis of Arabinosidase in Pichia pastoris
Dong-Sheng YAO ; Hui-Mei TAN ; Hui HUANG ; Da-Ling LIU ; Chun-Fang XIE ;
China Biotechnology 2006;0(04):-
SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.
2.Cardiac ischemia in type 2 diabetes mellitus rats induced by high sucrose and high fat diet and STZ treated.
Xue-Li YAO ; Jin WANG ; Wei-Fang ZHANG ; Xiao-Liang WANG ; Hui-Rong LIU
Chinese Journal of Applied Physiology 2014;30(2):137-140
OBJECTIVETo build a type 2 diabetes mellitus rat model with cardiac ischemia.
METHODSMale Wistar rats were fed high sucrose and high fat diet for four weeks and then injected with streptozoticin (STZ) (40 mg/kg .i.p.). The levels of fasting blood glucose and serum insulin were monitored every week. The body weights of rats were also measured every week. The blood levels of creatine kinase and lactate dehydrogenase (LDH) were measured following the electrocardiograph used BL-410 biological experiment system.
RESULTSThe serum insulin levels of diabetic rats were 4.05 ng/ml after four weeks high sucrose and high fat diet. The fasting blood glucose levels of diabetic rats were 17.9 mmol/L after injection. Compared with normal group, there was obvious change of S-T segment in the electrocardiograph of diabetic group at the fourteenth week. The levels of creatine kinase and lactate dehydrogenase in diabetic group significantly increased in comparison with those in normal group.
CONCLUSIONThe cardiac ischemia of diabetic rats model is suitable for investigating cardiac disease of diabetes mellitus.
Animals ; Creatine Kinase ; blood ; Diabetes Mellitus, Experimental ; physiopathology ; Diabetes Mellitus, Type 2 ; chemically induced ; physiopathology ; Diet, High-Fat ; adverse effects ; Dietary Sucrose ; adverse effects ; Disease Models, Animal ; L-Lactate Dehydrogenase ; blood ; Male ; Myocardial Ischemia ; physiopathology ; Rats ; Rats, Wistar ; Streptozocin
3.The Radiosensitizing Effect of Resveratrol on Hopypharyngeal Carcinoma Cell Line FADU and its Effect on the Cell Cycle.
Yuan SHAO ; Fang QUAN ; Hong-hui LI ; Xiao-bao YAO ; Qian ZHAO ; Rui-min ZHAO
Chinese Journal of Integrated Traditional and Western Medicine 2015;35(6):699-703
OBJECTIVETo study the radiosensitizing effect of resveratrol on hypopharyngeal carcinoma cell line FADU in vitro.
METHODSHypopharyngeal carcinoma cell line FADU was cultured in in vitro DMEM. Its inhibition on cell proliferation was detected using cytotoxicity test (MTT assay). The cell survival curve was drawn using clone formation to obtain sensitive enhancement ratio (SER). Changes of the cell cycle and cell apoptosis were analyzed using flow cytometry (FCM).
RESULTSResults of MTT showed the inhibition of resveratrol on FADU cells increased along with its concentrations (P < 0.05). Results of clone formation indicated the surviving fraction at 2 Gy (SF2) was 0.717 ± 0.062 in the irradiation group, and 0.426 ± 0.035 in the resveratrol plus irradiation group (with SER ranged 1.684 ± 0.178) with statistical difference (P = 0.007). Results of FCM showed that after radiation of 4 Gy radiation, cells at G2/M phase arrest increased, but cells at G1 decreased. After radiation of resveratrol for 24 h, cells at G1 decreased, but cells at G2/M phase and S phase arrest increased. When 4 Gy radiation combined resveratrol was used, cells at G2/M phase arrest significantly increased, but cells at G1 significantly decreased. The apoptosis rate was 1.94% ± 1.65% in the control group, 4.56% ± 0.92% in the irradiation group, 2.03% ± 1.46% in the resveratrol group, and 23.11% ± 7.22% in the resveratrol plus irradiation group. There was statistical difference between the resveratrol plus irradiation group and the rest 3 groups (P < 0.05).
CONCLUSIONResveratrol could enhance the radiosensitivity of hypopharyngeal carcinoma FADU cells in vitro possibly by inducing cell apoptosis and causing changes in the cell cycle distribution.
Apoptosis ; Carcinoma, Squamous Cell ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Head and Neck Neoplasms ; Humans ; Hypopharyngeal Neoplasms ; drug therapy ; Radiation Tolerance ; Radiation-Sensitizing Agents ; therapeutic use ; Stilbenes ; therapeutic use
4.Prevalence and influencing factors of dysmenorrhea among female college students
Chuanli PENG ; Yue ZHANG ; Hui LIANG ; Hongzhi HAN ; Fang YANG ; Yan YAO ; Weiying GUO
Chinese Journal of Health Management 2016;(1):50-54
Objective To investigate the prevalence and influencing factors of dysmenorrhea among female college students in Changchun city, so as to provide scientific basis for health promotion and effective preventive measurement. Methods Non-randomized convenience sampling and face to face interview were used to collect information from female college students aged between 17 and 25 years in 14 universities in Changchun. Chi-square test and logistic regression model were used to analyze influencing factors of dysmenorrhea. Results The average age of 1 071 subjects was 21.21 ± 1.83 years. The prevalence of dysmenorrhea was 86.55%. The proportion of mild dysmenorrhea among the subjects was 62.56%, followed by 33.01% with moderate dysmenorrhea and 4.43% with severe dysmenorrhea; 80.76% of subjects paid attention to keep warm in the daily life. Normal BMI, sleeping before 23 o'clock or between 23 to 24 o'clock, taking exercise frequently or everyday might be the protective factors of dysmenorrhea, and the OR values (95% CI) were respectively as 0.60 (0.37-0.97), 0.56 (0.37-0.84), 0.42 (0.22-0.78) and 0.63(0.42-0.97). Tension and the family history of dysmenorrhea might be the risk factors, and the OR values (95%CI) were respectively 1.63 (1.10-2.41), 4.84 (2.80-8.35). Conclusion The prevalence of dysmenorrhea is high among female college students. Lacking exercise, BMI less than 18.5 kg/m2, staying up late, tension and the family history of dysmenorrhea may be the influencing factors of dysmenorrhea among female college students.
5.Application of therapeutic communication in nursing of stroke patients
Hui LIU ; Zhihong TAN ; Jianling YAO ; Haishan YUAN ; Junying GAO ; Fang WAN
Modern Clinical Nursing 2015;(7):22-26
Objective To evaluate the effect of therapeutic communication on negative emotion and quality of life of stoke patients. Methods A total of 120 stroke patients were divided into the control group and the observation group equally. The former received routine nursing and the latter routine nursing plus therapeutic communication. After 4 weeks, they were evaluated by self-rating anxiety scale (SAS), self-rating depressive scale (SDS), trait coping style questionnaire (TCSQ) and the short-form health survey-36 (SF-36). Results After the intervention, the scores on anxiety, depression and negative coping in the observation group were lower than those in the control group (P<0.01), while the scores on positive coping and quality of life were higher in the observation group than those in control group (P<0.01). Conclusion Therapeutic communication can effectively improve stroke patients′negative emotion and quality of life.
6.Clinical distribution and drug resistance trend of Acinetobacter baumannii and analysis on its drug resistance in comprehensive hospital
Rongfeng YAO ; Juying SHEN ; Guoxiang XU ; Zhendong TAO ; Hui FANG ; Long XU
International Journal of Laboratory Medicine 2017;38(2):194-197
Objective This work was to study the distribution of Acinetobacter baumannii and analyze the change in the trend of its resistance,so as to provide experimental basis for clinical rational drug usage.Methods The identification of isolates were car-ried out by using VITEK-2 compact automatic identification system,and drug susceptibility testing was performed by disc diffusion method,the results were carried out according to the Standard of CLSI 2014 version for interpretation,and data analysis was per-formed with WHONET5.6.Results A total of 981 strains of Acinetobacter baumannii had been isolated from the samples in 2013 to 2015,most of them were isolated from respiratory samples accounted for 80.4%,followed by the urines and other body secre-tion.The distribution of this kind of bacteria mostly was 30.1% in ICU,departments of Emergency observation(16.0%),respirato-ry diseases(15.7%)and senior cadres(10.8%).The drug resistance of this kind of bacteria to polymyxin B and cefoperazone/sul-bactam was to a minimum of lower than 5.0%,and has relatively high sensitivity to amikacin and minocycline(<30.0%).And the resistance to other antimicrobials were higher than 30.0%,and the resistance to Nitrofurantoin and cefotaxime were higher than 94.0%.The most drug resistance of Acinetobacter baumannii in 2014 were lower than those in 2013,but the most drug resistance of Acinetobacter baumannii in 2015 were increased significantly as well as strains had been isolated from the samples.Conclusion The main source for isolation of Acinetobacter baumannii in this hospital mainly comes from sputum specimens of patients in ICU, departments of emergency observation,respiratory diseases and senior cadres.The high prevalence of antimicrobial resistance of this kind of bacteria should be prevented and controlled in nosocomial infection,and rational application of antimicrobial agents in order to reduce the spread of drug-resistant strains.
7.Orthogonal Experiment of Serum ContainingBuyang Huanwu Decoction in Influencing Apoptosisof PC12 Cells Induced by Oxygen-Glucose Deprivation
Jiping ZHANG ; June LIU ; Hui YAO ; Yongqi FANG ; Yuping HE ; Fengni WEN
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(3):469-474
Objective To investigate the protective effect of Buyang Huanwu Decoction ( BYHWD) on PC12 cell apoptosis induced by oxygen-glucose deprivation ( OGD) and to screen out the main ingredients of BYHWD on protecting PC12 cell from injury induced by OGD. Methods Seven herbs of BYHWD were selected as the influencing factors, and they were Radix Astragali, Radix Paeoniae Rubra, the carda part of Radix Angelicae Sinensis, Pheretima Asiatica, Rhizoma Chuanxiong, Semen Persicae, and Flos carthami. Two levels were designed for each factor, and the experiment was arranged according to L8 ( 27) orthogonal test table. Apoptosis of PC12 cells induced by OGD was detected by serum pharmacology method and flow cytometry techniques. With apoptotic rate as the evaluation index, visual analysis and variance analysis were used for the comparison of apoptosis of PC12 cells in the blank control group ( oxygen + sugary medium + blank serum) , model group (oxygen-glucose-free medium + blank serum) and drug groups (oxygen-glucose-free medium +serum containing BYHWD or the separate recipe of BYHWD ) . Results Compared with the blank control group, apoptotic rate was increased in PC12 cells of the model group, the difference was statistically significant ( P<0.01) . Compared with the model group, the apoptotic rate was decreased in BYHWD group, and the differences was statistically significant (P<0.01) . The results of orthogonal analysis showed that Semen Persicae, Flos carthami and Radix Paeoniae Rubra were the main active ingredients of BYHWD on depressing apoptosis of PC12 cell induced by OGD ( P<0.01). The apoptotic rate of serum containing the prescription with Radix Paeoniae Rubra was lower than that of the prescription without Radix Paeoniae Rubra. Conclusion BYHWD could inhibit OGD-induced PC12 cell apoptosis, and Radix Paeoniae Rubra is the main active herb on depressing apoptosis in BYHWD.
8.Gender differences of brain metabolic networks: a 18F-FDG PET study
Lei FANG ; Jianping AN ; Junfeng MAO ; Xuejiao CHEN ; Yuanwei XIE ; Hui ZHAO ; Wansheng LIANG ; Changshuai YAO
Chinese Journal of Nuclear Medicine and Molecular Imaging 2017;37(9):544-549
Objective To investigate the gender differences of glucose metabolic network in brains of healthy adults at resting state by 18F-FDG PET.Methods A total of 204 dextromanual,healthy individuals (104 males,average age:(53.45±11.51) years;100 females,average age:(54.11±12.09) years) were enrolled from June 2011 to June 2016 to construct brain metabolic networks.The nodal and global parameters,including clustering coefficient (Cp),characteristic path length (Lp) and betweenness centrality (Cb),were analyzed by graph theory.Permutation test with 1 000 repetitions was used.Results The brain metabolic networks derived from 18F-FDG PET data were with small-world properties in both male group and female group.Compared with Cb in females,Cb in males was significantly reduced in left postcentral gyrus,right angular gyrus and left temporal pole/middle temporal gyrus (permutation test,all P<0.05);and it was increased in left amygdala,left precuneus,right temporal pole/middle temporal gyrus and left inferior temporal gyrus (permutation test,all P<0.05).Comparing with the females,the male group had higher Cp and longer absolute Lp but without significant difference (permutation test,all P>0.05).Conclusions There are gender-related differences of topological structure in whole-brain metabolic networks.Gender should be considered as a covariate while designing experiments,accounting for cerebral metabolic data from normal control and experimental patients as well as making clinical decisions.
9.Therapeutic Efficacy of Acupuncture at Baihui (GV 20) and Shenting (GV 24) for Post-stroke Cognitive Impairment: A Systematic Review
Fang LIU ; Li-Qun YAO ; Jin-Hui CHEN
Shanghai Journal of Acupuncture and Moxibustion 2018;37(1):104-111
Objective To evaluate the clinical efficacy of acupuncture at Baihui (GV 20) and Shenting (GV 24) in treating post-stroke cognitive impairment (PSCI) by using the systematic review method.Method Via computer, Chinese Journal Full-text Database (CJFD), Wanfang, China Biology Medicine disc (CBMdisc), Chinese Science and Technology Periodical Database, PubMed, Foreign Evidence-Based Medicine (FEMB), the Cochrane Library were retrieved.Chinese Acupuncture and Moxibustion andShanghai Journal of Acupuncture-moxibustion were manually retrieved. Randomized controlled trials published before Jan 31st of 2013 on acupuncture at Baihui (GV 20) and Shenting (GV 24) in treating PSCI, both in Chinese and English, were collected. The required data were extracted, then were evaluated according to the criteria of Cochrane systematic review and underwent meta-analysis by using RevMan 5.0.Result Twenty-two clinical trials were finally recruited, including 1637 subjects. The meta-analysis showed that acupuncture at acupoints including Baihui (GV 20) and Shenting (GV 24) produced a more significant rehabilitation result compared to single rehabilitation training or medication. The comparison of Mini-Mental State Examination (MMSE) score showed: [WMD=3.37, 95%CI (1.70, 5.05),P<0.00001]; the comparison of P300 latency: [WMD=1.22, 95%CI (0.84, 1.59),P<0.00001]. Severe adverse reactions were not discovered. Conclusion Acupuncture at Baihui (GV 20) and Shenting (GV 24) can effectively improve the cognitive function of PSCI patients. However, the diagnostic criteria and evaluation indexes are expected to be unified and standardized, and the clinical trials on acupuncture intervening PSCI are required to be further improved methodologically.
10.Integrated DNA barcoding database for identifying Chinese animal medicine.
Lin-Chun SHI ; Hui YAO ; Li-Fang XIE ; Ying-Jie ZHU ; Jing-Yuan SONG ; Hui ZHANG ; Shi-Lin CHEN
China Journal of Chinese Materia Medica 2014;39(12):2155-2159
In order to construct an integrated DNA barcoding database for identifying Chinese animal medicine, the authors and their cooperators have completed a lot of researches for identifying Chinese animal medicines using DNA barcoding technology. Sequences from GenBank have been analyzed simultaneously. Three different methods, BLAST, barcoding gap and Tree building, have been used to confirm the reliabilities of barcode records in the database. The integrated DNA barcoding database for identifying Chinese animal medicine has been constructed using three different parts: specimen, sequence and literature information. This database contained about 800 animal medicines and the adulterants and closely related species. Unknown specimens can be identified by pasting their sequence record into the window on the ID page of species identification system for traditional Chinese medicine (www. tcmbarcode. cn). The integrated DNA barcoding database for identifying Chinese animal medicine is significantly important for animal species identification, rare and endangered species conservation and sustainable utilization of animal resources.
Animals
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DNA Barcoding, Taxonomic
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methods
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Databases, Nucleic Acid
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Eukaryota
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classification
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genetics
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Medicine, Chinese Traditional