1.Application of two dimensional electrophoresis,western blot and mass spectrum to screen markers of endometriosis
Hong ZHANG ; Yidong NIU ; Jie FENG ; Huifang GUO ; Xue YE
Journal of Peking University(Health Sciences) 2003;0(04):-
Objective:To find out markers of endometriosis. Methods: The two dimensional gel images of proteins extracted from eutopic endometrium from endometriosis patients and controls were analyzed by software Phoretix 2D,and the proteins expressed differently were identified primarily by query of data base.The proteins extracted from ectopic endometrium of ovarian endometriosis were transferred from two dimensional gel onto nitrocellulose membranes,followed by incubation with sera from women with and without endometriosis. Analyzed by MALDI-TOF-MS,the proteins hybridized differently were identified through their Peptide Mass Footprints. Results: Having compared the reproducible two dimensional gel images of proteins from eutopic endometrium of women with and without endometriosis,we obtained 11 proteins expressed differently.Through Western Blot technique,we found three proteins hybridized differently which were identified as vimentin,?-actin and ATP synthase ? subunit respectively. Conclusion: The protein expression spectra of eutopic endometrium from patients with endometriosis are significantly different from those of the controls,and the anti-endometrial autoantibodies against vimentin, ?-actin and ATP synthase ? subunit may be induced.
2.INHIBITION OF T-CELL APOPTOSIS BY TRANSFECTING ANTISENSE HUMAN Fas GENE AND ITS ROLE OF TUMOR THERAPY
Hong ZHANG ; Jie FENG ; Tianyun FU ; Xue YE ; Jingb ZHANG
Acta Anatomica Sinica 1953;0(01):-
Objective\ By inhibiting approach of apoptosis of T cells to observe if it could reduce the attacks of tumor cells upon lymphocytes and its possible role in tumor treatment. Methods\ The expressions of Fas/FasL genes in ovarian carcinoma cells were detected by using flow cytometry and RT\|PCR method.The construction of pcDNA3\|antisense Fas,the constructed vector was transfected into Jurkat cells with lipofectin,the change in expression of Fas gene was determined by flow cytometry.By means of Annexin\|V and MTT the effect of apoptosis in transduced Jurkat cells was investigated,and also using MTT cytotoxic test to investigate how 3AO cells kill Jurkat cells. Results\ Fas/FasL were expressed in 6 ovarian carcinomal cells.The expression level of Fas protein in Jurkat cells transduced with the constructed vector was decreased.Apoptosis was reduced in antisense Fas\|transfected Jurkat cells after anti\|Fas treatment. Conclusion\ FasL expression in ovarian carcinoma may be one of the reasons for ovarian carcinoma to escape immunosurveillance and attacking lymphocytes.Blocking Fas expression in lymphocytes can partially inhibit Jurkat cells apoptosis induced by anti\|Fas and reducing the attacks of tumor cells upon lymphocyte.\;
4.Application of MRI in severity grading and activity staging in thyroid associated ophthalmopathy
Qingshan HONG ; Jie SHEN ; Yaoming XUE ; Yanping CHEN ; Zhichun UN
Chinese Journal of Endocrinology and Metabolism 2010;26(3):215-216
The influence of MRI on severity grading and activity staging in thyroid associated ophthalmopathy (TAO) was evaluated.The results showed that MRI had a higher detection rate in abnormal extraocular muscle than NOSPECS scores(χ~2 = 19.37,P = 0.000).MRI had higher sensitivity,specificity,and accuracy in detection of active TAO than clinical activity score (100% vs 95.7%,88.2% vs 35.3%,96.9% vs 79.7%,respectively).Therefore,MRI improves early detection of atypical grade 4 cases.Signal intensity elevation in adipose suppressed T2WI is helpful to define active stage.
5.Combination of Thyroid Imaging-reporting and Data System and Ultrasound Elastography in the Differentiation of Benign and Malignant Thyroid Nodules
Jie XUE ; Xiaoli CAO ; Hong JIANG ; Zhibin WANG
Chinese Journal of Medical Imaging 2015;(5):351-355
PurposeTo evaluate the value of combined employment of thyroid imaging-reporting and data system (TI-RADS) and ultrasound elastography (UE) in the differentiation of benign and malignant thyroid nodules.Materials and Methods The clinical data of 174 patients with 232 thyroid nodules confirmed surgically or pathologically with puncture biopsy were retrospectively analyzed. All nodules were examined by ultrasound and UE. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of TI-RADS, UE and the combination of the two methods were calculated respectively, and the receiver operating characteristic curve (ROC) was drawn to evaluate the value of each of the three methods in differentiating benign and malignant thyroid nodules by comparison of area under curve (AUC).Results The diagnostic sensitivity, specificity and accuracy of UE (number of benign nodules: 155, number of malignant nodules: 77) were 88.9%, 91.8% and 90.9%, respectively; those of TI-RADS (number of benign nodules: 153, number of malignant nodules: 79) were 76.4%, 85.0% and 82.3%, respectively. The values measured by UE were significantly higher than those by TI-RADS (χ2=3.920 and 7.446,P<0.05). The sensitivity, specificity and accuracy measured by the combined use of the two methods (number of benign nodules: 155, number of malignant nodules: 77) were 93.0%, 93.7% and 93.5%, respectively, which were higher than either of the methods, and the difference was statistically significant (χ2=7.725, 6.450 and 13.728,P<0.05). The areas under the curve (AUC) were 0.833 and 0.812 respectively for UE and TI-RADS; the AUC for the combination of the two methods was 0.914, which had significantly higher diagnosis accuracy than that by any single method, and the difference was statistically significant (Z=1.95 and 2.55,P<0.05). Conclusion Compared with gray scale ultrasound, the combination of TI-RADS and UE has higher diagnostic sensitivity and accuracy for excluding malignancy in diagnose of thyroid nodules.
7.Fingerprint and spectrum-effect relationships on Tripterygium glycosides preparation.
Jie CHI ; Bing LIN ; Zhi-hong LIU ; Li-na YANG ; Xue-mei LIU ; Hong-tao SONG
China Journal of Chinese Materia Medica 2015;40(8):1479-1483
Tripterygium glycosides preparation which extracted from the traditional Chinese herb Tripterygium wilfordii (TWHY), was widely used to treat the autoimmune diseases. Previous works demonstrated that TWHF had potent anti-inflammatory and immunosuppressive properties. But the different quality and high incident rate of side effects of different manufactures inhibited its clinical application. Since TWHF had been generally known to play a therapeutical effect by synergism of multiple constituents, it was necessary to build the relationship between the HPLC fingerprint and bioactivity so as to ensure the quality safety and efficacy. The HPLC fingerprint showed that description and content of peaks from different manufactures were diverse. Only 11 common peaks were found. In this study, mice spleen cells stimulated by Con A were used to test the proliferation inhibition bioactivity of TWHF preparations, which were incubated with 30, 15, 7.5, 3.75, 1.88 and 0.94 mg x L(-1) TWHF preparations for 48 h. The results showed that mice spleen cells proliferation was inhibited by all TWHF preparations significantly compared with the control group, which suggested the TWHF preparations showed immune suppress activity. The TWHF preparations from 7 manufacture showed different IC50 value, which might belong to different contents which showed in the HPLC fingerprint. Moreover, a relationship between the HPLC fingerprint and the bioactivity were established to identify important constituents by grey relational analysis (GRA). The result showed that all the contents were relative with the IC50, especially No. 5 and 10 peaks, but No. 1 peak, which was proved to be triptolide, had few contribute to the inhibition of mice spleen cells proliferation. The study of relationship between the HPLC fingerprint and the IC50 by GRA could help to investigate mechanism of bioactive and provide an evidence for the quantification of multi-constituents.
Animals
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Anti-Inflammatory Agents
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analysis
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pharmacology
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Cell Proliferation
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drug effects
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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analysis
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pharmacology
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Glycosides
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analysis
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pharmacology
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Male
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Mice
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Mice, Inbred BALB C
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Spleen
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cytology
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drug effects
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Tripterygium
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chemistry
8.Regulation of a Highly Repeated GT Sequence from DCA1 Promoter of Dunaliella Salina in Salt-induction
Qing-Ju LUO ; Jie LI ; Hong-Xia YAN ; Xue-Jing LU ; Yu-Min LU ; Le-Xun XUE ;
China Biotechnology 2006;0(07):-
To identify whether a highly repeated GT sequence from DCA1 promoter from Dunaliella salina,which have been proved to be a salt-inducible promoter in our previous study,would be a salt-inducible regulation element,different primers were designed to amplify 6 different-length fragments of DCA1 promoter from D.salina by PCR.After these fragments were respectively inserted into the HindⅢ-BamH I sites of the vector pU?GUS,serial expression vectors containing the gus gene were generated.D.salina cells transformed with these recombinant plasmids by electroporation were grown in liquid media containing different concentrations of sodium chloride respectively.GUS enzyme activity was measured histochemically and fluorometrically.The results revealed that 3 fragments containing GT repeated sequence drove the external gus gene expression and the expression pattern of the gus gene was regulated by the concentrations of sodium chloride.Additionally,the 2 fragments without tandem GT sequence drove the gus gene expression,but the expression pattern of the gus gene wasn't regulated by the concentration of sodium chloride;Also,the upstream fragment of the tandem GT sequence wasn't able to drive the gus gene expression.In conclusion,the highly repeated GT sequence from the DCA1 promoter plays an important role in the salt-inducible regulation of DCA1 promoter from D.salina and might be a novel salt-inducible element.
9.Expression of chemokine-like factor 2 (CKLF2) mRNA in the rat myocardium after myocardial infarction
Yanjun GONG ; Tao HONG ; Jie JIANG ; Xue YU ; Dingfang BU ; Yong HUO ; We GAO
Journal of Peking University(Health Sciences) 2003;0(04):-
Objective: To investigate the expression of chemokine like factor 2 (CKLF2) mRNA in the rat myocardium after myocardial infarction(MI).Methods: In a rat model of MI, the myocardium surrounding the infarcted area was used for RNA preparation at different time points. After RT, competitive polymerase chain reaction amplification was performed to assess the expression of rCKLF2 mRNA. SAS Kruskal Wallis test was used for statistical comparisons. Results: The gene expression of rCKLF2 at mRNA level was significantly increased in the myocardium surrounding the infarcted area 3 days, 1 week, 2 weeks after infarction.Conclusion: It is possible that CKLF2 contributes to the pathophysiological process and needs to be further investigated.
10.The role of DCs in the immunoprotective effects of active immunization of elemene combo-tumor cell vaccine
Guangxia SHI ; Bing XUE ; Hong QU ; Lianying GUO ; Jie SHEN ; Zhenchao QIAN
Chinese Journal of Pathophysiology 2001;17(8):789-
In this experiment, Hca-F or L615 elemene combo-TCV (H-TCV and L-TCV), H-TCV lysates, corynebacterium parvum (CP) were used to immunize 615 and Balb/c inbred mice, and their splenic DCs were prepared and pulsed in vitro with tumor cell lysates (H or L) and TCV lysates (TH or TL). The capacities of DCs to stimulate the proliferation of syngeneic nonadherent spleenic cells were tested with MTT assay. The results showed that the splenic DCs from normal mice in vitro pulsed with TH or TL could induced syngeneic noradherent splenic cells to proliferate (P<0.01), while the H or L pulsed DCs could not. The splenic DCs from H-TCV or L-TCV or TH immunized mice re-pulsed in vitro with TH or TL exhibited stronger stimulating effects than the DCs from normal mice pulsed in vitro for the firth time pulsed with TH or TL (P<0.01 or P<0.05); The capacities of DCs to induce proliferation of syngeneic nonadherent splenic cells could be further enhanced by CP immunization, especially when were pulsed with TH (P<0.01). Normal inbred 615 mice were transferred with DCs pulsed with lysates of elemene TCV (TDCs) or pulsed with lysates of Hca-F tumor cells (HDCs) on day 7 before challenged with lethal dose of live Hca-F cells, significant adoptive immunoprotective effects were seen, with 61.6% tumor inhibition rate and 25% survival in TDC adoptive transfer group. This study indicated that DCs might play a role in the mechanisms of active immunization and pulsing DCs with lysate of elemene combo TCV and isolating DCs from elemene combo TCV immunized mice were useful methods for DCs vaccine preparation.