1.Immunohisochemical differential diagnosis of mesenteric fibromatosis, solitary fibrous tumors and gastrointestinal stromal tumors
Juanhong SHI ; Yan WANG ; Qingsi HE
Chinese Journal of Current Advances in General Surgery 1998;0(01):-
Objective: To study the immunohistochemical features and the differential diagnosis of mesenteric fibromatosis (MF), solitary fibrous tumors (SFT)and gastrointestinal stromal tumors (GIST). Methods: The expressions of CD117, CD34, ?-catenin, S-100, Desmin, Bcl-2 and Dog-1 of the three tumors were studied by immunohistochemistry method (S-P method). Results: By immunohistochemical staining detection, the expression of ?-catenin in six MF cases were positive, the positive expression rates of CD117 and Dog-1 in GIST cases were 85% and 92% respectively, and the positive expression rates of CD34 and Bcl-2 in SFT were 71% . Conclusion: MF, SFT and GIST can be identified by the combining applying of CD117,CD34 and ?-catenin through the immunohistochemistry method.
2.Determination of host bacterium genomic DNA content in DNA vaccine with digoxigenin-labeled probe
Yan HE ; Ke SHI ; Shuhan SUN ;
Academic Journal of Second Military Medical University 2000;0(08):-
The ribosomal DNA fragment was amplified with PCR utilizing synthetic primers with host bacteria chromosomal DNA as template. The resultant probes were labeled with Digoxigenin and were applied in the dot blot of DNA vaccine samples. The host bacteria genomic DNA in DNA vaccine against hepatitis B was less than 15 ng/?g. Digoxigenin labeled probes proved sensitive and reliable in determining genomic DNA.
3.Proteomics analysis of Sut melanocytes in response to xCT deficiency
Liming SHI ; Liping ZHAO ; Lu HE ; Wenhui YAN ; Haixuan QIAO
International Journal of Biomedical Engineering 2011;34(3):140-144
Objective To investigate the mechanism of Sut melanocytes growth inhibition in response to xCT -deficiency. Methods TTotal proteins were extracted from xCT-deficient Sut melanocytes and wild melanocytes, respectively, and were separated by 2-dimensional electrophoresis. Altered expression profile of proteins of Sut melanocytes was analyzed by PDQuest software and compared with that of wild melanocytes.Proteins with significant change were chosen to be identified by mass spectrometry and database query.Results Twenty proteins in Sut melanocytes altered significantly compared with wild melanocytes. Ten of the proteins were up-regulated, while the other tens were down-regulated. Four proteins from both up-regulated and down-regulated were identified respectively: up-regulated proteins were Tubulin alpha-1b, S100-A6,Nucleoside, S-formylglutathione, and down-regulated proteins were Calumenin,NDRG1 ,DPYSL2, 14kDa unknown protein. Conclusion The identification of the xCT-deficiency related proteins may provide supporting evidence for the mechanism research of Sut melanocytes' growth inhibition caused by xCT-deficiency.
4.Detection of integron structure and sequence of drug resistance gene cassette in clinical multidrug resistant Shigella dysenteriae strains
He YAN ; Minhua ZONG ; Lei SHI ; Yamasaki SHINJI
Chinese Journal of Infectious Diseases 2010;28(5):273-277
Objective Clinical strains of Shigella dysenteriae isolated from eastern parts of India in 1988, 1995 and 2002 were examined for the presence of class 1, 2 and 3 integrons which closely related with drug resistance and the distribution of resistance gene cassette in order to clarify the influence of integron system on drug resistance of Shigella dysenteriae. Methods Susceptibility to antimicrobial agents was tested by the disk agar diffusion method. Class 1, 2 and 3 integron genes (intI) were screened by polymerase chain reaction (PCR) in 16 clinical strains with drug resistance.The variable regions of gene cassette of positive strains were sequenced. Results All 16 isolates were resistant to at least 4 agents including β-lactams, aminoglycosides, tetracyclines, sulfonamides,chloramphenicols and quinolones. Class 1 integron gene was detected in 13 strains and all isolates carried class 2 integron which indicated that strains with two integron structures were detected simultaneously and class 3 integron was not detected. Class 1 integron inserted gene cassette was mainly blaara30 -aadA 1 family, conferring resistance to β-lactamase, spectinomycin and streptomycins isolates carried class 2 integron were mainly dfrAl-satl genes cassettes conferring resistance to methoxybenzyl aminopyrimidine and streptothricin, while dfrA\-sat\-aadA\ genes were present only in 4 isolates. Conclusions These data indicate that class 2 integrons are widespread in Shigella dysenteriae strains, and closely associated with multidrug resistance of Shigella.
6.DETECTION OF ORIENTIA TSUTSUGAMUSHI DNA FROM THE SPECIMENS OF BLOOD CLOT AND SPLEEN COLLECTED BOTH FROM EXPERIMENTAL INFECTED MICE AND FIELD RODENTS BY NESTED POLYMERASE CHAIN REACTION
Yansheng YAN ; Liang CHENG ; Shi HE ; Enshu YU
Chinese Journal of Zoonoses 2000;(6):39-41
AimTo evaluate the efficiency of gene amplification technique used in detecting the specimens colleted from rodents to identify natural epidemic foci of scrub typlus. MethodMice of Kunining strain were experimentally infected by a certain amount of Oriential tsutsugamushi. The specimens of blood clot and spleen from the infected animals were detected by nested polymerase chain reaction(nPCR)specific to O. T sutsugamush at the day 3,6 and 9 of post-infection. Then the technique was used for detection of samples collected from field. As an infected index ,the specimen was considered to be positive only if a 88-bp DNA fragment from Sta 58kDa gene of O. Tsutsugamushi could be produced. According to the study ,it was estimated whether or not that the sampling area is a natural epidemic focus of the disease. ResultsThe specimens of both blood clot and spleen from the mica at day 3 of post-infecction showed negative to the specific PCR product ,but positive when detected at day 6 and hereafter. Of 111 spleen samples from the field collections in the northwest of Fujian province,one was positive, and another positive sample was in the 29 blood clots from Jiangxi province. It is demonstrated that these areas have been the natural epidemic foci. Conclusion The nPCR method is of highly sensitive and specific to be used in the etiologic study on specimens from field rats.
7.Expression of miRNA-21 in colonic mucosa and peripheral blood CD4+ T cells of 49 patients with active inflammatory bowel disease
Yan SHI ; Caiyun MA ; Chong HE ; Zhanju LIU
Chinese Journal of Digestion 2015;35(12):830-833
Objective To investigate the role and significance of miRNA-21 in the pathogenesis and development of inflammatory bowel disease (IBD).Methods The expressions of microRNA-21 (miRNA-21) in colonic mucosa and peripheral blood CD4+T cells were analyzed by fluorescence quantitative polymerase chain reaction (PCR) in 26 patients with active Crohn's disease (CD), 23 patients with active ulcerative colitis (UC) and 19 healthy controls.The relative expression amount of miRNA-21 in colonic mucosal and peripheral blood CD4+ T cells in 11 patients with CD was compared before and after infliximab (IFX) treatment, and its correlation with tumor necrosis factor (TNF)-α was analyzed.t-test was performed for comparison between groups.The correlation between miRNA-21 expression in peripheral blood CD4+ T cells and serum TNF-α was analyzed by Spearman correlation analysis.Results The expression of miRNA-21 in colonic mucosa of CD patients and UC patients was 4.720 ± 0.397 and 4.993±-0.415, respectively, which were both higher than that of healthy control group (1.101±-0.284),and the differences were statistically significant (t =7.657 and 8.356, both P<0.01).The expression of miRNA-21 in peripheral blood CD4+ T cells of CD patients and UC patients was 3.254 ± 0.323 and 3.450±0.311, which were both higher than that of healthy control group (0.891 ±0.220), and the differences were statistically significant (t=6.007 and 7.098, both P<0.01).After 12 weeks of IFX treatment, simple endoscopic score for Crohn's disease (SES-CD) of CD patients was 8.019 ± 0.349,which was lower than that before treatment (21.502 ± 1.185), and the difference was statistically significant (t=10.910, P<0.01).The serum TNF-α level was 61.570±6.252, which was lower than that before treatment (173.500 ± 12.670), and the difference was statistically significant (t=7.926, P<0.01).The results of correlation analysis indicated that serum TNF-α level was positively correlated with miRNA-21 expression in peripheral blood CD4+ T cells (r =0.780 4, P < 0.01).Conclusions The increasing of miRNA-21 expression in IBD was closely correlated with the serum TNF-α level and severity of inflammation.miRNA-21 might be a new target for IBD treatment.
8.The spatial analysis of myocardium ultrastructure and observation of activity of mitochondrial enzymes
Shi-ying, ZHU ; Yan-he, ZHU ; Lian-bang, ZHAI
Chinese Journal of Endemiology 2009;28(3):288-290
Objective To perform a spatial analysis of myocardium uhrastructure and the activity of mitochondrial succinate dehydrogenase in sub-acute Keshan disease. Methods Myocardium samples were collected from the cases with sub-acute Keshan disease and non-myocarditis(control), and their ultrastructure was observed under electron microscope. The density of mitochondrion volume and cristal membrane and its volume were measured by a point-counting method, while mitochondrion volume was estimated by water displacement method, succinate dehydrogenase activity of mitochondrion by iron-copper method in sub-acute Keshan disease and non-myocarditis cases. Results The volume ratio of mitochondrion to the cell on myoeardium [(47.79±6.20)%], the area ratio of mitochondrion to sarcoplasm [(55.06±6.50) %], mitochondrion to myofibrils [(1.43±0,41)%], mitochondrion section area[(0.78±0.15)μm2], and ratio of the lesion of cristal membrane area to the matrix area and mitochondrion volume[(67.14±13.96)%, (44.62±13.44)%]in sub-acute Keshan disease group were obviously higher than those in control [(33.20±7.62)%, (38.07±9.43)%, (0.71±0.33)%, (0.44±0.07)μm2, (14.11± 12.51)%, (9.34±11.28)%; t = 3.75,7.93,6.61,36.40,52.65,37.51, all P < 0.05]. The volume ratio of myofibrils to cell[(34.52±5.12)%]and the area ratio of cristae mitochondria to matrix[(32.43±14.42)%]in sub-acute Keshan disease group was obviously less than those in control [(48.51±4.30)%, (86.04±12.37)%; t = 9.85, 53.46, both P < 0.05)]. Succinate dehydrogenase activity was negative in sub-acute Keshan disease group. Conclusions Myocardium ultrastructure changes in sub-acute Keshan disease including the increase of volume and areas of mitochondria and the damage of the cristal membrane in mitochondria. Succinate dehydrogenase activity is decreased or even disappeared.
10.CBR improves elder stroke patients' ability of activities of daily living
Yan WANG ; Xiang-dong XU ; Shi-ming HE ; Ming YANG
Chinese Journal of Rehabilitation Theory and Practice 2002;8(12):762-763
ObjectiveTo observe the effect of community-based rehabilitation (CBR) on older stroke patients in ability of activities of daily living (ADL).Methods50 older stroke patients were randomly divided into the rehabilitation group and control group. The rehabilitation group was treated with motor function exercise and ADL training, while the control group only took medicine. Two groups were evaluated with Barthel index before and after treatment. ResultsScores of Barthel index on the rehabilitation group were higher than that on the control group after treatment, and there was a significantly difference between two groups (P<0.01). Conclusions CBR has the significant effect on improving ADL in older stroke patients.