2.The clinical significance of anti-nuclear protein B23 in systemic sclerosis
Zhongqiang YAO ; Jun MA ; Xiangyuan LIU ; Guofeng WANG ; Mengxue YU
Chinese Journal of Rheumatology 2011;15(3):179-182
Objective To explore the clinical significance of anti-nuclear protein B23 antibody in systemic sclerosis. Methods Enzyme-linked immunosorbent assay was employed to detect the serum antinuclear B23 autoantibody. Mann-Whitney U test was used to compare the clinical and autoantibody profiles between SSc patients with B23 antibody and those without B23 antibody. Logistic regression analysis was employed to analyze the correlation between B23 antibody and clinical manifestations and autoantibody profiles. Results Mann-Whitney U test showed that, forced vital capacity (FVC) diffusion capacity of CO (DLco) in B23 positive SSc was significantly lower than that in B23 negative counterparts, pulmonary artery hypertension was more prevalent in B23 positive SSc patients. While anti-fibrillarin, anti-U1RNP, and antic entromere antibodies were more prevalent in B23 positive SSc. Multivariate logistic regression showed that anti-B23 antibody positivity was an independent risk factor for pulmonary artery hypertension in SSc (OR=123.92, 95%CI 26.67~575.66, P<0.01), and a protective factor for severe gastrointestinal involvement (OR=0.08, 95%CI 0.01 ~0.70, P<O.05). Logistic analysis showed that anti-B23 antibody was correlated with antifibrillarin (OR=11.50, 95%CI3.85~34.37, P<0.01) and anti-U1RNP antibodies (OR=3.43, 95%CI 1.01~11.63, P<0.05), and correlated with different degree of pulmonary artery hypertension. Conclusion The pulmonary artery pressure should be monitored closely in those SSc patients with a positive B23 antibody.
3.Effect of Kanglaite Injection on Proliferation and Apoptosis of Human Breast Cancer MCF-7 Cells
Jun CHEN ; Yan MA ; Yu LI ; Long QIN ; Li YAO
Chinese Journal of Information on Traditional Chinese Medicine 2014;(11):48-51
Objective To investigate effects of Kanglaite injection on proliferation, cycle and apoptosis of human breast cancer MCF-7 cells;To discuss its relevant mechanism. Methods Logarithmic growth phase cells were divided into control group and Kanglaite-treatment group (10, 20, 40μL/mL). Cells were cultured in RPMI-1640 for 24 h before drug treatment. The inhibition rate of Kanglaite injection on proliferation of human breast cancer MCF-7 cells was detected by MTT assay. Apoptosis and cell cycle of MCF-7 cells were detected by flow cytometry. Changes in cell nucleus were determined by Hochest staining assay. Protein expressions of Bcl-2 and Bax were detected by ELISA and Western blot. Results Kanglaite injection for 12 h, 24 h or 48 h resulted in a significant inhibition of MCF-7 cells proliferation (P<0.05, P<0.01);Compared with the control group, Kanglaite injection-treated cells showed increased percentage in G2/M and G0/G1 phases (P<0.001, P<0.01), but showed decreased percentage in S phase (P<0.01), and apoptosis rate increased (P<0.05, P<0.001). Kanglaite injection significantly decreased protein expression of Bcl-2, and enhanced protein expression of Bax of MCF-7 cells (P<0.01, P<0.001). Conclusion Kanglaite injection can inhibit the proliferation of human breast cancer MCF-7 cells, decrease cell cycle and induce apoptosis, the mechanism is related with decreasing protein expression of Bcl-2 and enhance the protein expression of Bax.
5.Surgical treatment for pulmonary carcinoid tumors: a single-center analysis of 62 patients
Feng YAO ; Jun ZHOU ; Chenxi ZHONG ; Yu YANG ; Heng ZHAO
Chinese Journal of Thoracic and Cardiovascular Surgery 2012;28(4):233-236
ObjectiveTo evaluate prognostic factors and the surgical results of pulmonary carcinoid tumors.Methods We retrospectively reviewed the medical records of 62 patients who were diagnosed as pulmonary carcinoid tumors between January 2000 and October 2010 at Department of Thoracic Surgery,Shanghai Chest Hospital.The following information was available for each of the 62 patients:age,sex,pathological type,and TNM stage.ResultsThere were no operative death.The 3-year and 5-year survival rates after surgery were 92.1% and 77.8%,respectively.Of the 62 patients,42 were diagnosed as typical carcinoid tumor,and among them,4 patients (8.3%) had lymph node metastases.Their 3-year and 5-year survival rates were 97.8% and 94.7%,respectively.The remaining 20 patients were diagnosed as atypical carcinoid tumor,and among them,6 patients (37.5%) had lymph node metastases.Their 3-year and 5-year survival rates were 84.4% and 58.8%,which were statistically significant compared with typical carcinoid tumor( P =0.0047 ).There was significant difference in survival rate between the patients with lymph node metastases and the patients without lymph node metastases (P =0.0048).CondusionThe main risk factors affecting survival rate of those patients who were diagnosed as pulmonary carcinoid tumors were pathological types and lymph node metastases.
6.A cellular model for the expression of the C-type lectin dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin: construction and functional analysis
Yu ZHANG ; Xu YAO ; Hanyan GU ; Baoxi WANG ; Jun LIU
Chinese Journal of Dermatology 2014;47(7):457-460
Objective To establish a cellular model for the expression of the C-type lectin dendritic cellspecific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN),and to provide a basis for the functional analysis of DC-SIGN.Methods The cDNA of DC-SIGN was obtained via PCR,and cloned into the eukaryotic expression vector porcine cytomegalovirus-enhanced green fluorescent protein (PCMV-EGFP) with EGFP at the N terminal of DC-SIGN.Then,the recombinant PCMV-EGFP-DC-SIGN plasmid was transfected into HEK293T cells followed by the detection of DC-SIGN expression using PCR,Western blot and flow cytometry.Confocal microscopy was performed to localize the expression of DC-SIGN-EGFP and visualize the recognization and internalization of the Derp2 allergen by DC-SIGN.Results The recombinant fluorescent fusion protein-expressing plasmid was successfully constructed.Both PCR and Western blot confirmed the expression of DC-SIGN.Flow cytometry showed that the expression of DC-SIGN was increased by approximately 50% in HEK293T cells transfected with the recombinant expression plasmid compared with those untransfected.As confocal microscopy showed,the green fluorescence-labelled DC-SIGN was located on the cell membrane,which could bind to the red fluorescence-labelled antigen Derp2 and internalize it into the cells.Conclusions The recombinant DC-SIGN-EGFP fusion protein is characteristically located on the surface of 293T cells,which can be recognized by the DC-SIGN-specific antibody and is capable of internalizing the allergen Derp2,and may serve as an ideal cell model for further studies on molecular function of DC-SIGN.
7.Synthesis and verification of herpes simplex virus envelope glycoprotein gC
Yu ZHANG ; Xu YAO ; Hanyan GU ; Baoxi WANG ; Jun LIU
Chinese Journal of Dermatology 2014;47(8):578-582
Objective To synthesize herpes simplex virus (HSV) envelope glycoprotein gC using gene engineering techniques,and to verify its expression.Methods Two separate parts of the HSV envelope glycoprotein gC,i.e.,GC-F and GC-R,were respectively synthesized.The GC-F and GC-R genes were synthesized,subcloned into the expression vectors pSumo-Mut (containing recognition sequences for endonucleases Stu1 and XhoI) and pCzn1 (containing recognition sequences for endonucleases NdeI and XhoI) respectively to form the recombinant plasmids pSumo-Mut-GC-F and pCzn1-GC-R.E.coli BL21 Arctic Express (DE3) cells were transformed with the two recombinant plasmids separately.Isopropyl thiogalactoside (IPTG) was used to induce the expression of target protein which was subsequently purified by nickel affinity chromatography.Finally,Western blot was performed to verify the reactivity of the synthesized protein with the sera of HSV-1-positive patients.Results Both GC-F and GC-R genes were synthesized by a total gene synthesis method.As sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) showed,the fusion proteins were mainly distributed in the sediment layer.The purity of GC-F and GC-R proteins was over 80% after purification by affinity chromatography.Western blot showed that both of the proteins were reactive with anti-HSV-1 antibody-positive sera.Conclusions Fusion expression vectors have been constructed for the gC protein,and IPTG successfully induces its expression.Moreover,the resulting proteins could react with anti-HSV-1 antibody-positive sera,and may serve as an ideal experimental material for next functional study.
8.Isolation,Characterization and Identification of SD Rats′ Oligodendrocyte Precursor Lineage Cells in vitro
jun, TANG ; lin, ZHONG ; yu-jia, YAO ; juan, CHEN
Journal of Applied Clinical Pediatrics 2006;0(23):-
Objective To obtain highly purified oligodendrocyte precursor lineage cells in vitro and make identification.Methods The oligodendrocyte precursors were separated from astrocyte by orbital shaker and further purified by differential adhesion,and finally cultured in chemically defined serum-free medium,with appended neurotrophin 2(N2),platelet-derived growth factor(PDGF),basic fibroblast growth factor(bFGF).Immunofluorescence assay was applied to identify the separated cells with A2B5,O4,O1 and glial fibrillary acidic protein(GFAP) antibodies.Results Over 95% of cultured oligodendrocyte precursor cells were obtained.The oligodendrocyte progenitors were A2B5 and O4 positive,immature oligodendrocytes were O4 and O1 positive while GFAP were negative.Conclusions Separation and purification by shaking and differential adhesion and chemically defined medium are suitable and effective to obtain highly purified oligodendrocyte precursor cells.Cell output will increase notabily and rest in immature phase by appending both N2,PDGF and bFGF.
9.Efficacy comparison between gemcitabine combined with nedaplatin and paclitaxel combined with nedaplatin in treatment of locally advanced nasopharyngeal carcinoma
Xuebing PAN ; Yu HE ; Junwei SHAO ; Mantian YAO ; Jun JIA
Cancer Research and Clinic 2017;29(6):394-397
Objective To investigate the effect of gemcitabine, paclitaxel combined with nedaplatin in treatment of locally advanced nasopharyngeal carcinoma. Methods 40 patients diagnosed as locally advanced nasopharyngeal carcinoma from May 2012 to August 2014 were randomly divided into observation group and control group. Patients in the observation group received gemcitabine plus nedaplatin chemotherapy, while those in the control group received paclitaxel plus nedaplatin chemotherapy. Then chemotherapy effect, adverse reactions, survival time and tumor marker contents were compared. Results The effective rate of the observation group was 65 % (13/20), which was higher than that (30 %, 6/20) of the control group (χ2 = 4.912, P< 0.05), however, there were no statistically differences in gastrointestinal reactions, bone marrow suppression, liver damage, kidney damage between groups (P> 0.05). Before chemotherapy, the SCCAg and CYFRA21-1 content of the control group were (2.30 ±0.31) ng/L and (18.27±2.19)μg/L, and the observation group were (2.34±0.27) ng/L and (18.48±2.25)μg/L, and there was no significant difference between the groups (P>0.05). After chemotherapy, the SCCAg and CYFRA21-1 content of the control group were (1.92±0.22) ng/L and (13.72±1.74) μg/L, and the observation group were (1.20 ±0.15) ng/L and (8.49 ±0.91) μg/L (P= 0.000). Conclusion Gemcitabine combined with nedaplatin chemotherapy can improve the chemotherapy effect and reduce tumor marker content, without increasing more adverse reactions, which is an ideal chemotherapy regimen for advanced nasopharyngeal carcinoma.
10.Signal mining for adverse drug reactions based on healthcare big data: methodology and applications
Xia ZHAO ; Yao CHEN ; Jun LIAO ; Feng YU ; Sheng LIN
Chinese Journal of Hospital Administration 2017;33(5):373-376
This paper presented the conventional methods for signal detection of adverse drug reactions (ADRs) and their applications, the research progress in ADRs signal mining based on healthcare big data, and briefed the methods and uses of ADRs prediction using machine learning technology in the era of healthcare big data.The conclusion was that deep learning, as a fast growing tool in machine learning, will become hotspot of research, expected to help with ADRs signal mining and rational clinical drug use.