1.The cloning and expression of an anti-Alzheimer's disease neuroprotective peptide gene humanin
Benyan LUO ; Min YUAN ; Min TANG ; Zhi CHEN
Chinese Journal of Neurology 2001;0(03):-
Objective To use gene engineering technique to express the humanin.Methods Humanin segment was synthesized and cloned into the prokaryocyte expression vector pGEMEX-1 and pet44a. The plasmid was transformed to E. coli BL21(DE3) by screening and identifying to obtain the high expressing positive clone.Results The humanin segment was cloned into vector successfully,and expressed efficiently with the inducement of IPTG. The molecular weight of the expressed recombinant protein of pGEMEX-1-humanin was about 28 kd while it was 63 kd in pet44a-humanin,and the expressed quantity of fusion protein was 40 percentage of the total bacterial protein while it was 20 percent in pet44a-humanin.Conclusion The humanin has been cloned and expressed successfully and the high efficiently expressed recombinant protein might contain the intact important structure domain of the humanin.
5.Determination of chromatographic fingerprint of Forsythia suspensa by pyrolysis-gas chromatography
Min YUAN ; Mingguang ZHANG ; Ting YANG ; Zhi ZENG ; Xiaoling HUANG ;
Chinese Traditional and Herbal Drugs 1994;0(01):-
Object To study comparatively the chromatographic fingerprint of Forsythia suspensa (Thunb ) Vahl in different producing aeras Methods Pyrolysis gas chromatography (PGC) was used Results The similar fingerprint of 13 samples (PGC) appeared The chromatographic overlap of them was over 89%, RSD
6.Construction of siRNA expression vector targeting IGF-1R and its ability to induce cell apoptosls in hmnan lung cancer cells
Ai-Qiang DONG ; Zhi-Yuan MA ; Min-Jian KONG ;
Chinese Journal of Thoracic and Cardiovascular Surgery 2003;0(04):-
Objective Insulin-like growth factor-1 receptor (IGF-1R),similar to insulin receptor,is one of the families of re- ceptor tyrosine kinases.,which has been found to be overexpressed in a variety of cancer.It is the main proliferation and survival sig- nal molecule in cancer cell and plays an important role in cancer growth and progress.Blocking signal transduction of IGF-1R by vari- ous strategies can suppress tumor growth and induce regression of established tumor.This study is to construct the siRNA expression vector targeting IGF-1R and to evaluate its ability to induce cell apoptosis in human lung cancer cells.Methods Two siRNA expres- sion vector,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 targeting IGF-1R,were constructed using pENTR/U6 vector,and a vector targeting hieiferase gene,pENTR/U6-shRNA-Iuc,was constructed as control.After vectors were transfected into A549 for 48h, knockdown of IGF-1R mRNA and protein and Akt phosphorylation were accessed,and DNA ladder and flow cytometry were used for cell apoptosis.Results siRNA expression vectors targeting IGF-1R were successfully constructed,which was confirmed by PCR and DNA sequencing,pENTR/U6-shRNA-1 and pENTR/U6-shRNA-2 demonstrated the expression were (22.1?2.5) % and (80.1? 3.9) % in IGF-1R mRNA level,(15.2?3.1)% and (47.1?4.1)% in protein level,respectively,compared with pENTR/U6- shRNA-luc.Suppression of IGF-1R by pENTR/U6-shRNA-1 blunted Akt phosphorylation,increased cell apoptosis induced by 3% ethanol,and retained 77.5 % A549 cells in the G0/G1 phase.Conclusion siRNA expression vector targeting IGF-1R can effectively suppress the expression of IGF-1R expression in A549.This study suggests that DNA vector-based RNAi has the potential to be effec- tive and practical cancer gene therapy strategy.
7.Fingerprint of Pogostemon cablin by pyrolysis-gas chromatography and its fuzzy cluster analysis
Mingguang ZHANG ; Min YUAN ; Peng YUAN ; Zhi ZENG ; Chunlan YAN ; Xiaoling HUANG ;
Chinese Traditional and Herbal Drugs 1994;0(08):-
Object To study comparatively the fingerprint of Pogostemon cablin (Blanco) Benth. in different producing area and investigate the effect of pyrolysis temperature and pyrolysis time on the fingerprint Methods Pyrolysis gas chromatography (PGC) and fuzzy cluster analysis were used Results Both similarity and charecteristics were found in the fingerprint of P. cablin in different producing area. The methodological evalulation showed that this method had a good reproducibility, RSD≤2.75% Conclusion This method is simple, rapid and accurate. It is suitable for the quality control of the Chinese materia medica with the important role of pyrolysis temperature and pyrolysis time
9.Anatomy, classification of intersinus septal cell and its clinical significance in frontal sinus endoscopic surgery in Chinese subjects.
Min WANG ; Fei YUAN ; Wei-wei QI ; Jye-yuan CHENG ; Xiao-pei YUAN ; Lin HAN ; Zhi-min XING
Chinese Medical Journal 2012;125(24):4470-4473
BACKGROUNDIntersinus septal cell (ISSC) is not a very uncommon frontal recess cell. But it is poorly described in literature. The clinical significance of this anatomic variant still remains unclear. The purpose of this study was to clarify the anatomy, classification of ISSC and its clinical significance in Chinese subjects.
METHODSWe prospectively identified ISSC in 200 consecutive subjects who had undergone computed tomography (CT) scans: 120 without frontal sinusitis (group 1) and 80 with frontal sinusitis (group 2). The ISSC was classified into two types: Type I ISSC communicated with frontal sinuses, type II ISSC communicated with frontal recess. The patients of frontal sinusitis had undergone functional endoscopic sinus surgery with the assistance of the classification of ISSC. Statistical analysis was performed to correlate the ISSC and its type to the presence of frontal sinusitis.
RESULTSThe ISSC was obvious when reviewing the coronal and axial CT scans. Of the 200 CT scans reviewed, ISSC were present in 90 (45%). Of the 120 scans in group 1, ISSC were present in 49 (41%), among which type I ISSC was in 22 (18%) and type II was in 27 (23%). Of the 80 scans in group 2, ISSC was present in 41 (51%), among which type I ISSC was in 16 (20%) and type II was in 25 (31%). There were no statistically significant differences about the frequency distribution of total ISSC, type I and II ISSC between group 1 and group 2.
CONCLUSIONSThe prevalence of ISSC was very high in Chinese patients. The classification of ISSC was helpful for surgeon to operate according to whether it communicated with frontal sinus or frontal recess. The type II ISSC could be relatively easily removed from frontal recess.
Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; Endoscopy ; methods ; Female ; Frontal Sinus ; anatomy & histology ; pathology ; Frontal Sinusitis ; classification ; diagnostic imaging ; surgery ; Humans ; Male ; Middle Aged ; Prospective Studies ; Radiography ; Young Adult
10.Restriction endonuclease digest - melting curve analysis: a new SNP genotyping and its application in traditional Chinese medicine authentication.
Chao JIANG ; Lu-Qi HUANG ; Yuan YUAN ; Min CHEN ; Jing-Yi HOU ; Zhi-Gang WU ; Shu-Fang LIN
Acta Pharmaceutica Sinica 2014;49(4):558-565
Single nucleotide polymorphisms (SNP) is an important molecular marker in traditional Chinese medicine research, and it is widely used in TCM authentication. The present study created a new genotyping method by combining restriction endonuclease digesting with melting curve analysis, which is a stable, rapid and easy doing SNP genotyping method. The new method analyzed SNP genotyping of two chloroplast SNP which was located in or out of the endonuclease recognition site, the results showed that when attaching a 14 bp GC-clamp (cggcgggagggcgg) to 5' end of the primer and selecting suited endonuclease to digest the amplification products, the melting curve of Lonicera japonica and Atractylodes macrocephala were all of double peaks and the adulterants Shan-yin-hua and A. lancea were of single peaks. The results indicated that the method had good stability and reproducibility for identifying authentic medicines from its adulterants. It is a potential SNP genotyping method and named restriction endonuclease digest - melting curve analysis.
Atractylodes
;
classification
;
genetics
;
DNA Restriction Enzymes
;
metabolism
;
DNA, Plant
;
genetics
;
Drug Contamination
;
Genotype
;
Lonicera
;
classification
;
genetics
;
Plants, Medicinal
;
classification
;
genetics
;
Polymorphism, Single Nucleotide