1.Screening of the protein interacting with inhibitor of differentiation 1′ using yeast two hybrid method
Siyuan JIA ; Xiangdong LUO ; Jie QI ; Yongyue SU ; Yu CHEN ;
Journal of Third Military Medical University 2003;0(19):-
Objective To obtain the protein interacting with inhibitor of differentiation1′(Id1′). Methods The recombinant bait plasmid pHybLex/Zeo Id1′ was constructed and transformed into yeast strain EGY48/ pSH18 34 to test pHybLex/Zeo Id1′ for non specific activation. Adult human lung cDNA libraries were screened to obtain true positive library plasmid. The true positive library clone was obtained by sequencing and basic local alignment sequence tool (BLAST). Results The recombinant bait vector, named as pHybLex/Zeo Id1′, was confirmed by sequencing. pHybLex/Zeo Id1′ was transformed into yeast strain EGY48/pSH18 34 and the transformants had no autonomously activated reporter genes. One true positive clone, obtained by screening of the adult human lung cDNA libraries, was confirmed to be Fyn by sequencing and BLAST. Conclusion Id1′ can interact with Fyn.
2.Qualitative analysis of the main chemical constituents of shuanghuanglian injection powder and their origin by HPLC-ESI/MS/MS spectrometry.
Qi-Zhi LUO ; Jia-Bo LUO ; You-Zhi WANG
Acta Pharmaceutica Sinica 2009;44(12):1391-1396
This paper is to report the analysis of the main chemical constituents of Shuanghuanglian injection powder and determination of their origin. The sample solution was analyzed by a Zorbax C18 column with a gradient mobile phase comprised of methanol and 0.25% acetic acid solution. Both UV and electrospray ionization mass spectrometry detector were used simultaneously, -Q1-scan detection mode was evaluated for the identification of the LC peaks. To analyze the mass spectrum of every LC peaks, 43 molecular mass from the ion chromatogram of Shuanghuanglian injection powder were identified and among them, structure of 20 compounds were elucidated, and the data were sorted to the three component herbs, separately.
Chlorogenic Acid
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analysis
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Chromatography, High Pressure Liquid
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methods
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Drug Combinations
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Drugs, Chinese Herbal
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administration & dosage
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chemistry
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isolation & purification
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Flavonoids
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analysis
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Forsythia
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chemistry
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Glycosides
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analysis
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Injections
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Lonicera
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chemistry
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Plants, Medicinal
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chemistry
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Powders
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Scutellaria
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chemistry
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Spectrometry, Mass, Electrospray Ionization
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methods
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Tandem Mass Spectrometry
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methods
3.Research on bone marrow mesenchymal stem cells in the treatment of severe acute pancreatitis
Yuqiang SHAN ; Liang LUO ; Qi XIE ; Yi WU ; Zhong JIA ; Huicheng JIN ; Guanghua FENG
Chinese Journal of Hepatobiliary Surgery 2013;19(9):711-715
Objective To investigate the mechanism of acute inflammatory response and tissue repair when rats accepted transplanted bone marrow mesenchymal stem cells (MSC) in severe acute pancreatitis (SAP).Methods A total of 40 rats were randomly divided into 4 groups which included the normal group (n=10),the model severe acute pancreatitis group (n=10),the transplanted bone marrow mesenchymal stem cells group (n =10),and the combination of bone marrow mesenchymal stem cells and granulocyte colony-stimulating factor (G-CSF) group (n=10).To cure the acute severe pancreatic injury caused by SAP,rats were injected with EDU-labeled MSCs and granulocyte colonystimulating factor (Gr-CSF).The conversion rate of MSCs to pancreatic cells or MSCs to endothelial cells was detected to assess the role of MSCs in tissue regeneration and repair.The expression of amylase,interleukin-6 (IL-6),and interleukin-10 (IL-10) in serum was detected to assess the role of MSCs in an acute inflammatory response.Results The concentrations of amylase and IL-6 were reduced and the concentration of IL-10 was increased in MSC and MSC+G-CSF groups after the onset of SAP.Flow cytometry showed a small amount of MSCs converting to endothelial or pancreatic cells,but the conversion rate was relatively low.Conclusions MSCs can play an important role in the antipre-release of inflammatory mediators,reducing acute immune response to control the acute inflammatory response in SAP.Moreover,MSCs can repair a damaged pancreas by converting into both pancreatic and endothelial cells.
5.The survey of rat density and flea index in households in villages having previous plague experience in Lianghe County, Yunnan Province in 2007
Jia-xiang, YIN ; Xing-qi, DONG ; Chun-hong, DU ; You-hong, ZHONG ; Xing-jian, SHI ; Jia-li, LUO ; Wei, LIANG
Chinese Journal of Endemiology 2009;28(4):417-419
Objective To evaluate the risk of plague occurrence via surveying and analyzing indoor rat density and flea index in natural villages having previous plague experience. Methods During August to September 2007, 30 natural villages experiencing previous plague were selected based on the surveillance data, and then all households were coded with numbers and 20 households in each village were randomly selected via computer. Cages and sticky papers were set in 600 selected households to capture rats and fleas. Rat density, flea prevalence, flea index and median were estimated. Results One hundred thirty-three Rattus flavipectus and 33 Suncus murinus were caught and averaged rat density was 2.8 rats per one hundred cage. nights (166/6000), the median was 5 rats each village. One hundred and one mice infected fleas, flea prevalence on rats was 60.8% (101/166), 296 Xenopsylla cheopis and 48 Leptopsylla segnis were collected. Rat flea index was 2.1 fleas per rat (344/166). A total of 315 dissociated flea was caught, average dissociated flea index was 0.026 fleas per sticky paper (315/11888). The median was 5.5 dissociated fleas per village. Of dissociated fleas, Ctenocephalides felis felis (205) and Xenopsylla cheopis (103) accounted for 97.8% (308/315). The proportion for species of the rat flea and the dissociated flea was different(Fisher test: P < 0.01). The rat flea was significantly associated with the rat density(r = 0.68, P < 0.01), but the dissociated flea was significantly associated with neither the rat density(r = -yield than fried wheat batter(χ2 = 5.59, P < 0.05). Conclusions In these villages having previous plague experience of Lianghe County, Rattusflavipectus was dominant species of indoor rats, Xenopsylla cheopis and Ctenocephalides felis felis were dominant species of rat flea and dissociated flea, respectively. Mengsong, Bangdu, and Tangjiatun village had potential risk of plague emergence.
6.Screening of AGGF1 interacting with inhibitor of differentiation-1.
Si-yuan JIA ; Xiang-dong LUO ; Jie QI
Journal of Southern Medical University 2007;27(9):1379-1381
OBJECTIVETo screen the proteins interacting with inhibitor of differentiation 1(Id1) using yeast two-hybrid analysis in adult human lung cDNA libraries.
METHODSThe coding sequence of Id1 was amplified by PCR and cloned into the bait plasmid. The recombinant bait vector pHybLex/Zeo-Id1 was verified by restriction endonuclease digestion before transformation into the yeast strain EGY48/pSH18-34, which was tested subsequently for reporter genes Leu2 and LacZ activation. The pHybLex/Zeo-Id1 plasmid and the cDNA library plasmid were sequentially transformed into the yeast strains and screened to obtain Leu2(+) and Leu2(+)LacZ(+) clones, with the false positive clones excluded using positive and negative controls, and the plasmid of the true positive clone was sequenced and blasted for homological analysis.
RESULTSSuccessful construction of pHybLex/Zeo-Id1 was confirmed by enzyme digestion. After transformation of pHybLex/Zeo-Id1 into EGY48/pSH18-34, no specific reporter genes Leu2 and LacZ activation was found. The pHybLex/Zeo-Id1 plasmid and the cDNA library plasmid were sequentially transformed into yeast strain, and 198 Leu(+) clones and 19 Leu(+)LacZ(+) double positive clones were obtained. After elimination of the false positive clones, one true positive clone was obtained, whose plasmid analysis by sequencing and blasting indicated high homology (99.5%, 556/559) to AGGF1 (an angiogenic factor with G-patch and FHA domains 1). AGGF1 expression was confirmed in the true positive yeast cells by Western blotting.
CONCLUSIONAGGF1 is confirmed to interact with Id1 by yeast two-hybrid analysis for screening adult human lung cDNA libraries.
Adult ; Angiogenic Proteins ; genetics ; metabolism ; Cell Proliferation ; Endothelial Cells ; cytology ; metabolism ; Gene Library ; Humans ; Inhibitor of Differentiation Protein 1 ; metabolism ; Lung ; cytology ; Plasmids ; genetics ; Protein Binding ; Two-Hybrid System Techniques
7.Purification and characterization of deoxyribonuclease from earthworm Eisenia foetida
Jianlin ZHANG ; Zhizhen LIU ; Xiaoyuan WANG ; Jianqiang YAO ; Jia LUO ; Jianhua WANG ; Lijun YANG ; Qi YANG ; Bo NIU
Journal of Peking University(Health Sciences) 2008;40(5):519-523
Objective:To purify a kind of deoxyribonuclease from earthworm Eisenia foetida (named earthworm DNase, EDNase) and study its characteristics. Methods: Acetone precipitation, ion-exchange chromatography, high performance liquid chromatography, SDS-PAGE, CapiUary electrophoresis isoelectric focusing and MALDI-TOP MS were used for the study. Results: This purified protocol improved 137-fold purification and 45.6 % recovery of enzyme activity. The molecular mass of EDNase was estimated to be 63 000. Mg2+ , Mn2+ and Ca2+ were strong inhibitors of EDNase, while Na+ slightly increasd the enzyme activity. The enzyme was completely stable in the pH range from 4. 4 to 5.2 and had a pH optimum of 4.8. The optimum temperature was 37℃ and the enzyme was stable up to 40 ℃. The pI of the enzyme was 6. 20. Km and Vmax for the enzyme were 1.52 g/L and 4. 89 mg/(mL ·min), respectively, with calf thymus DNA as substrate. The enzyme was able to degrade chromosemal DNA, linear λbacteriophage DNA as well as supereoiled plasmid DNA, but didn' t display any RNase activity. Conclusion: This kind of deoxyribonuclease possesses unique characteristics, which is different from the deoxyribonucleases which we have known before.
8.Polymorphism of vitamin D receptor gene FokI and susceptibility of prostatic cancer: a meta-analysis.
Jia CAO ; Xiaxi LI ; Qi HOU ; Rong LI ; Rongcheng LUO
Journal of Central South University(Medical Sciences) 2012;37(12):1215-1220
OBJECTIVE:
To quantitatively investigate the association between the polymorphism of FokI of the VDR gene and the susceptibility of prostatic cancer.
METHODS:
Databases of Pubmed, EMBase, CBM, CNKI, VIP, and Wanfang Data were retrieved from the date they formed till May 2011. All randomized controlled clinical trials which matched both the inclusive criteria and exclusive criteria were subjected to meta-analysis, conducted on Revman 5.0.0 software. Stata 11.0 software was employed to process Begg's test.
RESULTS:
Ten studies were included. Total sample cases were 8360, with 3749 cases in the patient group and 4611 cases in the control group, respectively. The quantitative analysis showed there were no significantly differences between the polymorphism of VDR FokI alleles and the susceptibility of prostatic cancer (allele F to f: OR=1.00, 95% CI=0.94-1.06, P=0.96; genotypes FF/Ff to ff: OR=1.04, 95% CI=0.93-1.51, P=0.48; genotypes FF to Ff/ff: OR=0.97, 95% CI=0.89-1.06, P=0.53). Though one research on Indian people indicated that allele F was a risk factor for prostatic cancer, in Begg's test we observed relatively high publication bias. The subgroup analysis showed there were no significantly differences between the polymorphism of VDR FokI alleles and the susceptibility of prostatic cancer (allele F to f, white race: OR=0.91, 95% CI=0.88-1.02, P=0.17; yellow race: OR=1.09, 95% CI=0.95-1.24, P=0.22; Indian: OR=1.91, 95% CI=1.30-2.81, P=0.0009).
CONCLUSION
VDR FokI allele F might be a protective factor for European and American Caucasians.
Alleles
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Deoxyribonucleases, Type II Site-Specific
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genetics
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Genetic Predisposition to Disease
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Humans
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Male
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Polymorphism, Single Nucleotide
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Prostatic Neoplasms
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genetics
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Receptors, Calcitriol
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genetics
9.Curative Effect of YU Yun Pulse-feeling-based Acupuncture Therapy for Treatment of Middle-late Liver Cancer
Yin Ze LIU ; Bo Hai ZHANG ; Ying LUO ; De Min XIE ; Ying Qian DONG ; Kang Wei ZHOU ; Qi Jia LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2018;35(1):66-69
Objective To investigate the curative effect of YU Yun pulse-feeling-based acupuncture therapy for the treatment of middle-late liver cancer. Methods A total of 60 middle-late liver cancer patients were divided into control group and treatment group by stratified randomization method,30 cases in each group. The control group was given integrated Chinese and western medicine therapy according to the clinical pathway, including anti-cancer treatment such as vascular interrention,molecular targeted therapy,chemotherapy,radio therapy, and focal ablation therapy,as well as chinese medicine treatment based on disease differentiation and syndrome differentiation. And the treatment group was given YU Yun pulse-feeling-based acupuncture therapy on the basis of treatment for the control group. The two groups received 12-week treatment, and then their curative effects were compared. Results The treatment group had better effect on increasing the survival rate, prolonging survival time, improving the scores of clinical symptoms, stabilizing tumor size, and increasing the scores of Karnofsky Performance Status (KPS)than the control group,the difference being significant (P < 0.05 or P <0.01). Conclusion YU Yun pulse-feeling-based acupuncture therapy exerts certain curative effect for the treatment of middle-late liver cancer.
10.The status on care and nutrition of 774 children staying in rural areas while parents were in towns
Xi-Chun PENG ; Jia-You LUO ; Kuan-Bao YAO ; Ru-Shan HU ; Qi-Yun DU ; Ming-Yuan ZHU
Chinese Journal of Epidemiology 2008;29(9):860-864
Objective To understand the status on care and nutrition of children living in the rural areas (so called 'left-behind' children) while their parents were seeking for jobs in the urban areas. Methods Cross-sectional study was employed in this investigation.The group of‘left-behind' children (n=774) and the comparison group (n=774) were identified.The ascertainment methods mainly included questionnaire,anthropometries measurements,food-frequency,and laboratory examination.Results be poorly attended (X2=6.671,P=0.036;X2=15.053,P=0.001 ).Mothers who chose to work outside of their households would tend to choose bottle-feeding or decrease the duration of breastfeeding for their infants (X2=5.051,P=0.031;t=-7.201,P=0.001).The intake of milk and bean products in children were obviously lower than that seen in the control group (t=-2.150,P=0.032 ;t=-2.054,significant difference when comparing with the control group (X2=4.560,P=0.033 ).Conclusion communities,parents and extended families would facilitate more attention and effective intervention programs to improve the situation.