2.Screening of Alzheimer's disease-associated gene expression changes through peripheral blood monocytes extracted gene microarray analysis in patients with Alzheimer's disease
Li TIAN ; Juan FENG ; Quan SHI ; Weishuang XUE ; Weiyu TENG
Chinese Journal of Geriatrics 2016;35(12):1267-1270
Objective To screen the amyloid protein-β(Aβ)degradation-associated genes through peripheral blood monocytes(PBMC)-extracted gene microarray analysis in patients with Alzheimer's disease(AD).Methods PBMC were isolated from blood of elderly AD patients versus age-matched healthy individuals(control).The cDNA(mRNAs)were analyzed using gene microarray.And real-time quantitative polymerase chain reaction detection and enzyme activity analysis were used to verify the primary outcome of gene chip.Results The expression of cathepsin D mRNA in peripheral blood monocytes was 104.70±15.96 in AD patients as compared with the control group 49.86±5.19,and the activity of cathepsin D was (22 620 ± 1 389) RFU in AD versus (32 210 ± 2 284) RFU in control (both P<0.05).Conclusions The results suggest that the decreased levels of cathepsin D could be the stage markers related to the pathophysiology of AD process.Based on the microarray data,we select cathepsin D genes for further study.
3.Exploration of the correlation between Chinese medicine syndrome types of verrucous gastritis and the pressions of gastric mucosal hypoxia-inducible factor-1alpha as well as downstream molecules.
Yu-feng BU ; Li-juan CHU ; Hui-juan SHI
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(10):1326-1330
OBJECTIVETo study the expressions changes of gastric mucosal hypoxia-inducible factor-1alpha (HIF-1alpha) and downstream molecules [such as vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2)] of verrucous gastritis (VG) patients of different Chinese medicine (CM) syndrome types and their clinical significance.
METHODSTotally 94 VG patients were assigned to Gan-Wei disharmony group (GWDG, 28 cases), the damp-heat in Pi-Wei group (DHPWG, 17 cases), the blood stasis in Wei-collateral group (BSWCG, 20 cases), and the insufficiency of Pi-yang group (IPYG, 29 cases). Another 30 patients with chronic mild non-active superficial gastritis patients accompanied with negative Hp infection were recruited as the control group. The Hp infection was detected using 14C-labeled urea breath test. The expressions of HIF-1alpha, VEGF, and COX-2 in the gastric mucosal tissue were detected using immunohistochemical EnVision two-step test.
RESULTSThe positive Hp infection rate in VG patients was 37.23% (35/94 cases). The positive Hp infection rate in patients of DHPWG (76. 47%) was significantly higher than the other three groups (32.14% in GWDG, 31.03% in IPYG, and 20.00% in BSWCG, P < 0.01). The expressions of HIF-1alpha and COX-2 in VG patients of different syndrome types were obviously higher than those of the control group (P < 0.01, P < 0.05). Of them, the expression of HIF-1alpha was the highest in BSWCG and the expression of COX-2 was the highest in DHPWG. The expression of VEGF was higher in DHPWG and IPYG than in the control group and the GWDG (P < 0.01, P < 0.05).
CONCLUSIONSThe expressions of HIF-1alpha, VEGF, COX-2, and Hp infection showed certain changes in VG patients of different syndrome types. The expression of HIF-1alpha was the strongest in BSWCG. The expressions of VEGF and COX-2 as well as Hp infection were the highest in DHPWG. All showed the specificity of CM syndromes.
Adult ; Aged ; Cyclooxygenase 2 ; metabolism ; Female ; Gastric Mucosa ; metabolism ; pathology ; Gastritis ; diagnosis ; metabolism ; pathology ; Helicobacter Infections ; diagnosis ; metabolism ; pathology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult
5.Quality standard study on Mori Cortex liquid extract.
Mao-feng LIU ; Mei-ping XIE ; Lan LI ; An-qi LU ; Jian-gong SHI ; Su-juan WANG
China Journal of Chinese Materia Medica 2015;40(10):1850-1854
A reasonable and practicable quality standard was developed for mori liquid extract from different sources by TLC, HPLC and fingerprint technology. In TLC method, the compounds were separated on polyamide film using glacial acetic acid-water (1: 3) as mobile phase at a UV wavelength of 365 nm. All qualified samples had the spots of the same color as the control herb and substance. The RP-HPLC method was used to determine the content of mulberroside A with mobile phase of methanol-water (25: 75) at a wave-length of 326 nm. The mulberroside A was in good linear with a regression equation of Y = 46.965X (r = 0.999 6) in the range of 4.6 - 228 mg x L(-1). In 14 batches of samples, the mulberroside A in 4 batches of them was less than 0.5 g x L(-1), and was more than 2.0 g x L(-1) in the other batches. It was suggested that the content limit of mulberroside A should be no less than 1.5 g x L(-1). The HPLC fingerprints were evaluated by the similarities. It has found that the similarities of different mori liquid extracts were very low and the chemical diversity of mori cortex was the major factor of similarity. Moreover, the process impact was minimal. Thus the fingerprint was not included in this quality standard.
China
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Chromatography, High Pressure Liquid
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Disaccharides
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chemistry
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isolation & purification
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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standards
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Morus
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chemistry
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Quality Control
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Stilbenes
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chemistry
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isolation & purification
6.Pathogens in expressed prostatic secretion and their correlation with serum prostate specific antigen: analysis of 320 cases.
Shu-Xia WANG ; Jia-Ming ZHANG ; Kai WU ; Juan CHEN ; Jian-Feng SHI
National Journal of Andrology 2014;20(8):715-718
OBJECTIVETo investigate the pathogenic infection and its drug resistance in expressed prostatic secretion (EPS) and its correlation with serum PSA, and provide some evidence for the systematic and normalized diagnosis and treatment of prostatitis.
METHODSThree EPS swabs were collected from each of the 320 prostatis patients following measurement of the serum PSA level, 1 for bacterial culture and identification, 1 for detection of Mycoplasma and drug sensitivity, and the other for examination of Chlamydia trachomatis antigen by colloidal gold immunoblot.
RESULTSTotally 244 strains were isolated from the 320 EPS samples, including 188 bacterial strains (dominated by Staphylococcus and sensitive to vancomycin or linezolid) and 44 Mycoplasma and Chlamydia strains (mainly Ureaplasma urealyticum and susceptible to josamycin or doxycycline). The serum PSA level was significantly higher in the pathogen-positive than in the pathogen-negative group ([6.98 +/- 0.56] microg/L vs [2.32 +/- 0.12] microg/L, P < 0.05).
CONCLUSIONProstatitis may lead to the elevation of the serum PSA level and the pathogens involved vary in their resistance to different antibacterial spectrums. Therefore, appropriate and individualized antibiotic therapy should be selected according to etiological diagnosis and the results of drug sensitivity test.
Adult ; Humans ; Male ; Middle Aged ; Prostate ; microbiology ; secretion ; Prostate-Specific Antigen ; blood ; Prostatitis ; blood ; microbiology ; Young Adult
7.Problems and countermeasures for communication equipment supportability of newly established field medical unit
feng Lei SHI ; zhao Feng FU ; hua Wei HE ; Juan LUO
Chinese Medical Equipment Journal 2017;38(11):118-120
Objective To explore the problems and countermeasures for the communication equipment supportability of newly established field medical unit. Methods The characteristics of the communication equipment assigned to the field medical unit were introduced, and the problems of the equipment were summarized during 2-a exercise, examination, training and etc of the field medical unit, then some countermeasures were put forward accordingly. Results The communication equipment had problems in low utilization rate, training, operation procedure and safety, and the countermeasures were brought out from the aspects of emphasis from the director, personnel training, technical performances of the communication equipment and etc. Conclusion The communication equipment needs improving to support the commanding system of the field medical unit although it behaves better than the original equipment.
8.Establishment and optimization of detection of the hepatitis B immune complexes.
Li-Feng XU ; Juan WANG ; Jing XU ; Nan SHI ; Gang WU
Chinese Journal of Experimental and Clinical Virology 2007;21(3):291-293
OBJECTIVETo establish and optimize methods to detect the immune complexes (IC) of hepatitis B virus directly.
METHODSA C1q solid phase ELISA, mouse anti-HBs MAb solid phase ELISA and the complement consumption assay were established to detect the IC and these methods were optimized.
RESULTSAll the three methods were highly sensitive, specific and reproducible. The C1q used for coating tended to lose its activity easily at room temperature. Although strict requirements are needed for the raw and processed materials for complement consumption assay and the process of manipulation is complex, it can quantitatively detect IC. Comparing to the C1q solid ELISA and complement consumption assay, the mouse anti-HBs MAb solid phase ELISA has its own merits: convenience and stability.
CONCLUSIONMouse anti-HBs MAb solid phase ELISA is the best way to detect IC directly.
Complement C1q ; analysis ; Complement Fixation Tests ; methods ; Enzyme-Linked Immunosorbent Assay ; methods ; Hepatitis B ; blood ; Humans ; Reproducibility of Results
9.Value of the hypointensive internal septation on MR for the differentiation between benign and malignant breast lesions
Pei-Hong GAO ; Bin ZHAO ; Shi-Feng CAI ; Guang-Bin WANG ; Hong-Juan PENG ; Shao-Ling LIU ;
Chinese Journal of Radiology 2001;0(07):-
Objective To investigate the diagnostic value of internal septation for differentiating benign from malignant breast lesions.Methods A total of 26 patients were included in the study,in which 12 patients had 20 lesions of breast carcinoma and 14 patients had 25 lesions of fibroadenoma diagnosed either pathologically or clinically.The differential diagnoistic value of the hypointensive internal septation was analyzed.Results The signal intensity of fibroadenomas and malignant lesions on T_2-weighted fat- suppressed images could be classified as iso- to hyper- intensity,hypointensity and mixed intensity. According to the signal intensity classification,there were 5,11 and 4 cases in patients with breast carcinoma respectively,while 11,10,4 cases in patients with fibroadenoma respectively.There was no statistical difference in the distribution between the two patient groups(?~2=1.764,P=0.414).The shape of fibroadenomas and malignant lesions could be classified as irregular、roundish or lobulated.According to the morphological classification,there were 12,7 and 1 case in patients with breast carcinoma respectively, while 1,7,17 cases in patients with fibroadenoma respectively.There was statistical difference in the distribution between the two patient groups(?~2=23.262,P=0.000).The typical features of fibroadenomas were as follows:lobulated shape,hypointensive internal septations on T_2-weighted or postcontrast images. The diagnostic sensitivity of the three imaging features for fibroadenoma was 68%(17/25),52%(13/25), and 72%(18/25)respectively;and the diagnostic specificity was 95%(19/20),90%(18/20),95% (19/20)respectively.Conclusion The internal septation is a rather specific sign for diagnosis of fibroadenomas.
10.Construction of eukaryotic recombinant expression plasmids with glyceraldehydes-3-phosphate dehydrogenase and cysteine protease inhibitor gene of periodic Brugia malayi and its expression in HeLa cells
Xiao-jun, LIU ; Xiao-feng, GUO ; Sai-nan, ZHANG ; Shi-juan, LU ; Hao, FANG ; Bang-sheng, XU ; Zheng, FANG
Chinese Journal of Endemiology 2011;30(4):371-375
Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.