1.Cloning and Characterization Analysis of 60S Ribosomal Protein L10a Gene from Chaetomium globosum
Zhi-Hua LIU ; Qian YANG ;
Microbiology 1992;0(01):-
The sequence of Neurospora crassa(XP_322380)and Gibberella zeae PH-1(EAA76971)ribosomal protein gene were subjected to local tBlastn searching against the Chaetomium globosum ESTs datebase.The 765 bp full length cDNA encoding 60S ribosomal protein L10a gene was obtained.The open reading frame was 654 bp and encoded 217 amino acids.The protein molecular mass was 23.9 kD.The BlastP analysis revealed that amino acids sequence of ribosomal protein L10a gene from C.globosum shared 89% high similarity with N.crassa and 78% low similarty with Ustilago maydis.The cDNA and deduced amino acid sequence of 60S ribosomal protein L10a gene were accepted by GenBank(accession numbers: AY669070,AAT74578).
2.Carrier erythrocytes and its application in targeting chemotherapy
Yang YANG ; Baorui LIU ; Xiaoping QIAN
Journal of Medical Postgraduates 2003;0(11):-
Erythrocytes, the most abundant cells in human body with excellent physiologic and morphologic characteristics, have been exploited extensively for their potential applications as drug delivering microspheres for several potential applications, including intravenous slow release of therapeutic agents, enzyme replacement therapy and drug targeting therapy. A series of methods has been tested successfully for the improvement of targeting to reticuloendothelial system.
3.Extraction and identification of exosomes in follicular fluid from patients with polycystic ovary syndrome and isolation and detection of miRNAs in exosomes
Qian YANG ; Lanxin LIU ; Hefeng HUANG
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(8):1085-1089
Objective · To extract and identify exosomes in follicular fluid from patients with polycystic ovary syndrome in order to determine the existence of exosomes in follicular fluid, to isolate and extract miRNAs in exosomes, and to detect relative expression of miRNAs. Methods · Exosomes in follicular fluid were collected with membrane affinity chromatography and their size and morphology were observed with the transmission electron microscope. Exosome protein markers CD63 and CD81 were detected with flow cytometry. miRNAs in purified exosomes were extracted and expressions of miR-125b, miR-19b, and miR-222 were measured with TaqMan real-time PCR. Results · Exosomes in follicular fluid were circular or elliptic under the transmission electron microscope with diameters of around 30-100 nm. They had complete membrane structure and contained low density matter. Flow cytometry showed that CD63 and CD81 were positively expressed in exosomes. Real-time PCR detected expressions of miR-125b, miR-19b, and miR-222. Conclusion · Exosomes can be collected in follicular fluid from patients with polycystic ovary syndrome. Transmission electron microscopy and flow cytometry can be used to identify exosomes in follicular fluid. miR-125b, miR-19b, and miR-222 can be detected in exosomes.
4.Clinical Efficacy of Xiaoer Chiqiao Qingre Granule for Child Wind-heat Common Cold Fever(Stagnant Type)
Ruijie ZHANG ; Yuanhui LIU ; Qian YANG
China Pharmacy 2005;0(24):-
OBJECTIVE:To observe the curative effect of Xiaoer chiqiao qingre granule in treating child common cold fever (stagnant type).METHODS:A total of 118 cases with child common cold fever(stagnant type) were randomly assigned to receive Xiaoer chiqiao qingre granule (treatment group,n=60) or ribovirin tablet,Ertangganmao granule(control group,n=58) tid for 3 days.The antipyretic time, clinical effective rate, and improvement in clinical symptoms were compared between the two groups. RESULTS: There outcome indexes including the antipyretic time,the clinical effective rate,and the improvement in clinical symptoms in the treatment group was significnatly better than in the control group(P
5.Optimization of Extracting Process of Feining Mixture with Orthogonal Design of Multi-indexes
Dakai YANG ; Zhihui LIU ; Fang QIAN
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(10):-
Objective To optimize the extracting process of the mixture of Feining.Methods Orthogonal design was used to optimize extracting process.The content of glycyrrhizicac,Ephedrine and Pseudoephedrin,and the extractive rate of water and n-butyl alcohol were indexes.Results The optimum water-extracting factors were 8 times of water,refluxing and extracting for 1 h,for 2 times.Conclusion The optimal extraction process is precise and repeatable,which can be used in industrial production.
6.Association between clara cell protein 10 and airway insufflation pressures in guinea pig asthma models.
Qian GONG ; Ao LIU ; Weikang YANG
Chinese Journal of Practical Internal Medicine 2001;0(04):-
0.05).The CC10 levels in lung homogenate of the allergic asthma group were significantly decreased than that of control group(P0.05).Conclusion The CC10 levels in lung homogenate is decreased,and has negative correlation with AIPs,which suggests that CC10 may take part in the onset of asthma,and work as an important endogenous anti-inflammatory factor.
7.Cloning and identification of human gene 3 transactivated by nonstructural protein 5A of hepatitis C virus
Yan LIU ; Qian YANG ; Jun CHENG
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective To screen and clone the target genes transactivated by nonstructural protein 5A (NS5A) of hepatitis C virus (HCV). Methods The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-NS5A and pcDNA3.1(-) empty vector, respectively. Suppression subtractive hybridization (SSH) method was employed to analyze the differentially expressed DNA sequence between the two groups. The obtained sequences were searched for homologous DNA sequence from GenBank. The new gene with no homology with known genes in this database was confirmed, and electric polymerase chain reaction was conducted for cloning the full-length DNA of the new gene and in conjunction with Kozak rule and the terminus of polyadenyl signal sequence. The reverse transcription PCR (RT-PCR) was used to amplify the new gene from mRNA of HepG2 cell as the template. The coding sequence of the new gene was deduced according to the nucleotide sequence. Results A new gene with unknown function was named as NS5ATP3. The nucleotide sequence of the NS5ATP3 gene and its corresponding amino acid have been determined, which contained 1 572nt and 524aa. The sequence of the NS5ATP3 gene was deposited into GenBank, with the accession number AF529364. Conclusions NS5ATP3 gene transactivated by HCV NS5A protein was cloned and identified successfully by combining molecular biological technology and bioinformatics technique. These results will pave the way for the study of the molecular mechanism of the transactivating effects of HCV NA5A protein and the development of new therapy for chronic hepatitis C.
8.Peroxynitrite aggravates myocardium injury in acute myocardial infarction during ischemia-reperfusion
Lin LIU ; Qian FAN ; Xinchun YANG
Chinese Journal of Interventional Cardiology 1996;0(04):-
Objective To investigate the changes of peroxynitrite(ONOO-)in acute myocardial infarction during ischemia-reper fusion.Methods Thirty patients with actute myocardial infarction(single coronary arterial stenosis of left anterior descending in proximal-middle segment) and twenty-four healthy cases were enrolled as the control in our research.Peripheral blood were taken at the time of 24 h and 72 h after primary percutaneous coronary intervention.The patients were further divided into 2 groups according to their serum cTnI level(mild myocardium injury group with cTnI 0.05).Conclusion Plasma peroxynitrite(ONOO-) is related to myocardium injury during ischemia-reperfusion in actue myocardial infarction which suggest that peroxynitrite may aggravate myocardium injury in acute myocardial infarction during ischemia-reperfusion.
9.Long-term Curative Effective analyse of Open Mastoidectomy With Tympanoplasty(OMT)and Combined Approach Tympanoplasty(CAT)
Yang LIU ; Jianjun SUN ; Jin QIAN
Journal of Audiology and Speech Pathology 1998;0(02):-
Objective To discuss the surgical interference of chronic suppurative otitis media and it's long-term results.Methods In 113 cases of mastoidectomy with tympanoplasty followed up for 3 years,We discuss the results of recurrence and hearing levle of the open-mastoidectomy with tympanoplasty(OMT)and combined approach tympanoplasty(CAT).Results There is no significant difference between OMT(improved 12 dB HL)and CAT(improved 9.5 dB HL)in the improved hearing threshold leve.The recurrence rate of OMT was 5.8%,but the CAT was 24.4%,the difference is significant.Conclusion The effect of OMT is better than that of CAT for the long-term resulls in these cases.
10.Cloning, Sequence Analysis and Proeukaryotic Expression of Peroxisomal Membrane Protein Allergen Gene from Chaetomium globosum
Zhi-Hua LIU ; Qian YANG ; Li-Ming YANG ;
China Biotechnology 2006;0(04):-
The full length cDNA encoding peroxisomal membrane protein from Chaetomium globosum was cloned using RACE technology and the sequence in cDNA library of C. globosum in GenBank ( Accn: BP099709). The 747bp full length cDNA encoding peroxisomal membrane protein allergen (pero) gene was assembled with 412bp 3'and 508bp 5'RACE products. The open reading frame was 501 bp encoding 166 amino acids. The molecular weight of the protein was 17. 5kDa and its theoretical isoelectric point was 5.75. The pero gene was amplified using specific primers of cDNA 5'and 3'untranslated region, sequence analysis indicated that the gene have 3 exons and 2 introns. ClustalX analysis revealed that amino acids sequence of pero gene from C. globosum and Neurospora crassa shared 83% high similarity. To construct pET28a-pero expressive plasmid, pero gene was inserted into pET28a expressive vector. Escherichia coli BL21 transformed by pET28a-pero plasmid was induced with IPTG. The protein expression was analyzed with SDS-PAGE. A 21kDa pero fusion protein representing the pero gene was expressed in recombinant E. coli BL21. The sequences of cDNA,DNA and deduced amino acid of the pero gene from C. globosum were submitted to GenBank (Accn: AY555771, AY584753,AAS66898).