1.Expression of mRNA for MCP-1 and CCR2 in cerebral tissue of rats with acute alcoholism.
Chun-yang XU ; Shuang LI ; Kun SHAO ; Rui-ling ZHANG ; Wei HAO
Chinese Journal of Applied Physiology 2011;27(3):274-379
Alcoholism
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metabolism
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Animals
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Brain
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metabolism
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Chemokine CCL2
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genetics
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metabolism
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Male
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RNA, Messenger
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genetics
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metabolism
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Rats
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Rats, Sprague-Dawley
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Receptors, CCR2
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genetics
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metabolism
2.Screening Citrinin Mutants from the Transformants Library of Monascus ruber M-7 by Agrobacterium-mediated DNA transfer
Yue-Di DING ; Yan-Chun SHAO ; Yi-Ping XU ; Fu-Sheng CHEN ;
Microbiology 1992;0(04):-
200 citrinin mutants were screened with the inhibition zone method from the transformants library of Monascus ruber M-7 by Agrobacterium-mediate DNA transfer, which contains more than 5,000 transformants.Then 53 mutants, whose citrinin contents ranged from 0.04?g/g to 154.57?g/g in the red fermented rice (RFR), were achieved by high performance liquid chromatography (HPLC).Color values of RFR prepared by these mutants were also detected.The results showed that there was a positive correlation between the citrinin content and the color value among the mutants.These results provide materials and research bases for ferrther studying the relationship between the production of citrinin and pigment of Monascus ruber at molecular level.
3.Detection of Salmonella spp.,Escherichia coli and Staphylococcus aureus by Multiplex PCR
Yi-Ping XU ; Wei CHENG ; Yan-Chun SHAO ; Fu-Sheng CHEN ;
Microbiology 1992;0(06):-
According to DNA sequences of the invA gene of Salmonella spp.,the phoA gene of Escherichia coli and the nuc gene of Staphylococcus aureus,three pairs of oligonucleotide primers were designed and synthesized to amplify the special DNA sequences by multiplex PCR. Moreover,the reaction conditions of multiplex PCR were optimized. The results showed the multiplex PCR using the three pairs of primers produced specific amplicons of expected sizes,284bp for Salmonella spp.,622bp for Escherichia coli,484bp for Staphylococcus aureus. The optimized reaction conditions followed as the concentration of primer 40nmol/L for Salmonella spp.,40nmol/L for Escherichia coli,80nmol/L for Staphylococcus aureus,2.4mmol/L Mg 2+ ,200?mol/L dNTP,1.5U Taq DNA polymerase,anneal temperature from 55.0℃ to 57.4℃. Under the condition,the detection limits for DNA template were 10.2pg,10.2pg and 102.0pg for Salmonella spp.,Escherichia coli and Staphylococcus aureus,respectively. The whole process could be completed within 4h. The multiplex PCR assay was a specific,sensitive,rapid and reliable method for detecting Salmonella spp.,Escherichia coli and Staphylococcus aureus,which establish important foundation for simultaneous detection for these three bacteria in food.
4.Construction and immunoscreening of cDNA library of Armillifer agkistrodontis nymphs
Ling-ling, ZHANG ; Jia-xu, CHEN ; Shao-hong, CHEN ; Jian, GUO ; Yu-chun, CAI
Chinese Journal of Endemiology 2012;31(6):599-603
Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.
5.Inhibitory effect of docetaxel on proliferation of human lens epithelial cells
Shao-ling, YI ; Bin, SHI ; Wan-wen, LI ; Li-jian, XU ; Chun-shun, ZHAO
Chinese Journal of Experimental Ophthalmology 2011;29(1):32-37
Background Some drugs with inhibitory effect on the proliferation of lens epithelial cells have a limiting application in clinic because of their adverse response.To screen the effective and less side-effect drug for supressing LECs growth is very inportant for the prevention and treatment of after cataract.Objective This study was to explore the effects of docetaxel on LECs growth and compare its role with epirubicin hydrochloride,pirarubicin hydrochloTide and rahitrexed.Methotis Immortalized human LECs line (SRA01/04) were cultured and passaged.Different concentrations of docetaxel,epirubicin hydrochloride,pirarubicin hydrochloride and rahitrexed were added into the medium respectively for 24.48 and 72 hours.The proliferation of LECs was detect by M1Yr.Flow cytometry analysis Was used to analyze the influence of different concentrations of docetaxel on cellular cycle at 48 hours after addition of docetaxel,and Annexin V-FITC/PI marking method was used to assesse the apoptosis of LECs under the action of docetaxel.Expression of bcl-2 protein in LECs Was evaluated by Westeru blot. Result The growth rate of LECs Wag 100%in 8-519 pmol/L doeetaxel groups with the normal cell shape.Majority of abnormal cells and low growth rate were found in 66 nmoVL docetaxel group at 48 and 72 hours.The IC50 of docetaxel was lowest in 48 and 72 hours in docetaxel group in comparison to epirubicin hydrochloride and pirarubicin hydrochloride. However,no evident inhibition on LECs growth in 23.22-523.56 μmol/L of raltitrexed.At 48 hours,the percentage of LECs in G2/M phase increased as the asccnte of concentration of docetaxel,showing a significant difference among 4 groups(F=2633.05,P<0.01).The percentage of early apoptotic cells increased to 22.4%(χ2=20.00,P<0.01) and 27.9%(χ2=42.68,P<0.01)from normal control 3.1% at 48 hours after LECs exposed to 8.3 nmol/L and 266 nmol/L docetaxe.The expression of bcl-2 protein in LECs was obviously weakened after addition of docetaxel,especially 8.3 nmol/L docetaxel group. Conclusion Docetaxel,epirubicin hydrochloride and pirarubicin hydrochloride can inhibit the proliferation of human LECs in vitro.But there is no supression on LECs growth inraltitrexed.Docetaxel is proved to have a strongly arrested effect on the proliferation of LECs in comparison with epirubicin hydrochloride and pirarubicin hydrochloride and play its role at concentration-and time-dependent manner.
6.Simultaneous determination of seven bioactive constituents in Smilacis Glabrae Rhizoma by high-performance liquid chromatography.
Shuo XU ; Ming-ying SHANG ; Guang-xue LIU ; Feng XU ; Feng-chun LI ; Xuan WANG ; Shao-qing CAI
China Journal of Chinese Materia Medica 2015;40(3):469-479
This study is to develop an HPLC method for the simultaneous determination of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoisoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in Smilacis Glabrae Rhizoma. Samples of Smilacis Glabrae Rhizoma, Heterosmilacis Chinensis Rhizoma and Heterosmilacis Yunnanensis Rhizoma were separated on an Agilent Zorbax SB-C18 column with gradient elution of acetonitrile-0.05% phosphoric acid at a flow rate of 1.0 mL · min(-1). The detected wavelength was set at 230 nm and the column temperature was maintained at 35 °C. The contents of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in 84 Smilacis Glabrae Rhizoma samples were in the range of trace-1.381, trace-9.913, trace-3.673, 0.6706-27.08, trace-1.181, trace-4.833 and trace-2.754 mg · g(-1), respectively. The established method was used for analysis of common adulterants. The results demonstrated that the contents of (-)-epicatechin in Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma were 0.01163-0.06007 mg · g(-1) and 0.01583-0.08585 mg · g(-1), respectively, while the other six constituents were not detected. The method is simple and accurate, and can be used for the quality control of Smilacis Glabrae Rhizoma. The constituents of Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma are significantly different from Smilacis Glabrae Rhizoma, and they are not suitable for using as Smilacis Glabrae Rhizoma.
Chromatography, High Pressure Liquid
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methods
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Drugs, Chinese Herbal
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analysis
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Liliaceae
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chemistry
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Rhizome
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chemistry
7.Social participation and needs for rehabilitation services of the disabled in Guangdong Province HUANG
feng DONG ; Xi CHEN ; Ai-hua LIN ; Shao-zhen CHEN ; Ming-xu JIANG ; Zhi-ming YANG ; Chun-guang GONG ;
Chinese Journal of Physical Medicine and Rehabilitation 2010;32(3):208-211
Objective To explore the relationship between social participation and needs for rehabilitation of the disabled in Guangdong Province and to make a proposal for developing the rehabilitation strategies. Meth-ods The data of the Second National Sample Survey of Disabled Persons in Guangdong Province was used in this study. Ranked data analysis was made with the sub-items of the social participation assessment and the main needs of the disabled individuals. Results Significantly statistical differences were revealed with regard to the constitu-ent ratio of needs for rehabilitation services among people with different degrees of difficulties in social participation caused by hearing and visual impairments as well as physical and mental disabilities. No significant difference was found in terms of the constituent ratio of rehabilitation needs among those with difficulties in speech and those with psychiatric diseases. The major rehabilitation needs focused on medical service, assistive apparatus support and functional trainings. Conclusions The rehabilitation needs were different among different categories of disabled persons. Rehabilitation services should be provided accordingly.
9.Processing environmental stimuli in paranoid schizophrenia: recognizing facial emotions and performing executive functions.
Shao Hua YU ; Jun Peng ZHU ; You XU ; Lei Lei ZHENG ; Hao CHAI ; Wei HE ; Wei Bo LIU ; Hui Chun LI ; Wei WANG
Biomedical and Environmental Sciences 2012;25(6):697-705
OBJECTIVETo study the contribution of executive function to abnormal recognition of facial expressions of emotion in schizophrenia patients.
METHODSAbnormal recognition of facial expressions of emotion was assayed according to Japanese and Caucasian facial expressions of emotion (JACFEE), Wisconsin card sorting test (WCST), positive and negative symptom scale, and Hamilton anxiety and depression scale, respectively, in 88 paranoid schizophrenia patients and 75 healthy volunteers.
RESULTSPatients scored higher on the Positive and Negative Symptom Scale and the Hamilton Anxiety and Depression Scales, displayed lower JACFEE recognition accuracies and poorer WCST performances. The JACFEE recognition accuracy of contempt and disgust was negatively correlated with the negative symptom scale score while the recognition accuracy of fear was positively with the positive symptom scale score and the recognition accuracy of surprise was negatively with the general psychopathology score in patients. Moreover, the WCST could predict the JACFEE recognition accuracy of contempt, disgust, and sadness in patients, and the perseverative errors negatively predicted the recognition accuracy of sadness in healthy volunteers. The JACFEE recognition accuracy of sadness could predict the WCST categories in paranoid schizophrenia patients.
CONCLUSIONRecognition accuracy of social-/moral emotions, such as contempt, disgust and sadness is related to the executive function in paranoid schizophrenia patients, especially when regarding sadness.
Adolescent ; Adult ; Asian Continental Ancestry Group ; European Continental Ancestry Group ; Executive Function ; Facial Expression ; Female ; Humans ; Male ; Middle Aged ; Schizophrenia, Paranoid ; psychology ; Young Adult
10.Effects of CASI on myocardial ischemia-reperfusion arrhythmia in rats.
Li WANG ; Xiao-Feng YU ; Shao-Chun QU ; Hua-Li XU ; Da-Yuan SUI
China Journal of Chinese Materia Medica 2007;32(20):2174-2177
OBJECTIVETo observe the protective effect of compound acanthopanax senticosus injection (CASI) on myocardial ischemia-reperfusion arrhythmia in rats.
METHODThe myocardial ischemia-reperfusion model was induced by 30 min coronary occulusion and 60 min reperfusion in openchest anesthetized rats. The changes of arrhythmia with electrocardiogram lead II, the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), the contents of malondialdehyde (MDA) and Ca2+ in myocardium were determined.
RESULTIn rats treated by CASI (in a dosage of 25, 50 and 100 mg x kg(-1) femoral vein infusion at 30 min after coronary occulusion), the incidence of myocardial ischemia-reperfusion ventricular arrhythmias, for instance the ventricular tachycardia (VT) and ventricular fibrillation (Vf), was effectively prevented, the appearing time of arrhythmia was delayed and the duration of arrhythmia was shortened, while the elevated ST segment lowered as well. At the same time, the contents of myocardial Ca2+ and MDA were decreased significantly as well as the activities of myocardial SOD and GSH-Px increased markedly.
CONCLUSIONCASI is of protective effect on myocardial ischemia-reperfusion arrhythmia, which may be related to scavenging the oxygen free radicals and Ca2+ overload formed during reperfusion.
Animals ; Anti-Arrhythmia Agents ; isolation & purification ; pharmacology ; Arrhythmias, Cardiac ; drug therapy ; etiology ; physiopathology ; Calcium ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Electrocardiography ; Eleutherococcus ; chemistry ; Female ; Ginsenosides ; isolation & purification ; pharmacology ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Myocardial Reperfusion Injury ; complications ; Myocardium ; metabolism ; pathology ; Panax ; chemistry ; Phytotherapy ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Saponins ; isolation & purification ; pharmacology ; Superoxide Dismutase ; metabolism