1.Isolation and identification of tumor-like stem cells from human nasopharyngeal carcinoma cell line
Xiaofang GUAN ; Zhong WEN ; Congxiang SHEN
Medical Journal of Chinese People's Liberation Army 2001;0(12):-
Objective To investigate the expression of CD133 in human nasopharyngeal carcinoma cell line SUNE,and to detect proliferative and differential ability of CD133+ tumor cells in vitro,and to identify tumor-like stem cells in SUNE.Methods Immunocytochemical staining method and flow cytometry were used to detect the expression of putative tumor-initiated cell marker CD133 in nasopharyngeal carcinoma cell line SUNE,and the isolation technique with immunomagnetic beads was applied to purify CD133+ cells.The proliferative ability of CD133+ tumor cells in vitro was estimated by MTT assay,and the proliferative ability of CD133+,CD133-and control SUNE cells were statistically compared.Finally,the immunocytochemical staining method and flow cytometry were used to detect differential ability of CD133+ tumor cells in vitro.Results CD133 was positively expressed only in 0.35% of the cells in nasopharyngeal carcinoma cell line SUNE.In the serum-free medium RPM I1640,the UV optical density of the CD133+ tumor cells enriched with immunomagnetic beads was 0.317?0.007,0.370?0.002 and 0.558?0.004 at 3,5 and 7 days,respectively.Compared with CD133-cells and control SUNE cells,CD133+ cells showed increased proliferative capacity(P
2.Phytohaemagglutinin stimulates the proliferation of peripheral blood mononuclear cells and expression of secretory cytokines
Ding WANG ; Bing SONG ; Xuan ZHONG ; Xiaofang SUN ; Yong FAN
Chinese Journal of Tissue Engineering Research 2014;(23):3707-3714
BACKGROUND:Phytohemagglutinin (PHA) can stimulate the peripheral blood mononuclear cells (PBMCs) into cellcycle, and cause their immune activation, which is a common immune proliferation model. However, the role of non-PBMC ingredient of peripheral blood is unclear, as wel as the expression of endothelial cells related cytokines. OBJECTIVE:To study the effect of whole blood culture and PBMCs alone culture with PHA on the PBMC proliferation and apoptosis, expression of inflammatory cytokine and endothelial cellsecreted cytokine markers. METHODS:Morphological changes of PBMCs separated from normal karyotype human peripheral blood individual y cultured with or without PHA were observed. The PBMCs were col ected by whole blood culture or PBMC separated culture. mRNA was extracted for the fluorescence quantitative RT-PCR, which was applied to detect the cellproliferation, apoptosis, and expression of inflammatory cytokine and endothelial cellsecreted cytokines. The statistic analysis was used for the significance explication. RESULTS AND CONCLUSION:PBMCs alone cultured ere different from those undergoing whole blood culture. The PHA could up-regulate the gene expression of Ki67, proliferating cellnuclear antigen, Caspase 3, interferon-γ, tumor necrosis factor-βand interleukin-6, but down-regulate Protein C. This indicted that PHA could promote the proliferation and apoptosis of PBMCs and up-regulate the expression of inflammatory cytokines, but down-regulate the expression of endothelial cells secreted coagulation cytokines.
3.Evaluating of left atrial remodeling and predicting the efficacy of radiofrequency catheter ablation with atrial fibrillation by speckle tracking imaging
Xiaofang ZHONG ; Weibin HUANG ; Jing WANG ; Jun GUO
Chinese Journal of Ultrasonography 2015;(5):382-386
Objective To evaluate left atrial function dynamic changes before and after radiofrequency catheter ablation (CA)with persistent atrial fibrillation (AF)and predict the maintain sinus rhythm of postoperative atrial fibrillation by speckle tracking imaging.Methods The study was including 45 cases underwent CA with persistent AF(preoperative 1 week,postoperative 24 hours,3 months,6 months)and 25 normal cases.Left atrial function and strain rate were analysed.These index were measured and calculated:the left atrial diameter index (LADI),left atrial area index (LAAI),left atrial maximum volume index (LAVImax)and left atrial ejection fraction(LAEF),left atrial wall strain rate of left ventricular systolie (SRs),left atrial wall strain rate of left ventricular early diastolie (SRe),left atrial systolic strain rate(SRa) and the average of left atrial wall each phase strain rate.Results ① LADI,LAAI,LAVImax were significantly increased;LAEF,SRs,SRe were decreased significantly;SRa were disappeared with AF compared with the control group.②LADI,LAAI,LAVImax decreased gradually,LAEF,SRs,SRe,SRa were gradually increased with the extension of the follow-up time.③Left atrial average SRs predicted maintenance of sinus rhythm with atrial fibrillation after operation of CA(odds ratio=0.607,95% CI=0.417-0.819,correction of P =0.017)was an independent predictor.Conclusions Speckle tracking imaging can quantitative evaluate left atrial function with persistent AF after CA.Left atrial global reservoir function predicts the efficacy of CA.
4.Preliminary study of the relationship between the ischemic mitral regurgitation mechanism and geometric structure of the mitral valve in the papillary muscle dysfunction
Xiaofang ZHONG ; Jing WANG ; Maolong SU ; Xiaoyang HUANG
Chinese Journal of Ultrasonography 2014;(11):925-929
Objective To study the relationship between ischemic mitral regurgitation and geometric angles of the mitral valve leaflets in patients with coronary artery disease and papillary muscle dysfunction by transesophageal echocardiography(TEE) ,and to evaluate anatomy and pathophysiology mechanicm of the ischemic mitral regurgitation with left ventricular papillary muscle dysfunction .Methods A total of 84 subjects were enrolled in this study .All of the subjects were divided into group A (healthy volunteers constituted the control group ,n =40) ,group B (chronic inferior or posterior myocardial infarction patients with papillary muscle dysfunction group ,n =44) ,group C(patients from group B after mitral valvuloplasty surgery ,n = 20) .The relationship between mitral regurgitation and geometric angles of the mitral valve leaflets and mitral annulus were studied by TEE .Results Geometric angle between the mitral valve leaflets and mitral annulus were significantly larger in group B ,and geometric angles of the mitral valve leaflets and mitral annulus was positive correlated with degree of mitral regurgitation .Degree of mitral regurgitation significantly reduced in group C ,no significant difference was found at the geometric angles of the mitral valve leaflets and mitral valve annular compared with group A .Conclusions The degree of mitral regurgitation were highly correlated with geometric angles of the mitral valve leaflets and mitral valve annular in patients with left ventricular papillary muscle dysfunction caused by coronary heart disease .Repair of the mitral valve leaflets surgery can significantly reduce geometric angles of the mitral valve leaflets and mitral valve annular ,and reduce the degree of mitral regurgitation significantly .
5.Study on the relationship between TK gene regulated by enhanced suicide gene vector and telomerase activity in nasopharyngeal carcinoma cells
Congxiang SHEN ; Zhong WEN ; Yuhong QIAN ; Shaofeng MU ; Xiaofang GUAN
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2010;(4):168-173
Objective:To explore the relationship between TK gene expression regulated by enhanced suicide gene vector and telomerase activity in nasopharyngeal carcinoma cells.Method:The reformed reconstructed enhanced vector, pGL3-basic-EGFP-TK-hTRETp-CMV enhancer, and hTERT mono-promoter vector, pGL3-basic-EGFP-TK-hTRETp(as controls), were transfected into telomerase(+) nasopharyngeal carcinoma 5-8F cell lines,telomerase(+) human breast cancer MCF-7 cell lines and telomerase(-) normal vascular endothelium cell lines respectively. TK gene green fluorescent protein was observed by fluorescence microscope. The expression of TK gene mRNA was measured by the real-time fluorescent quantified PCR and the telomerase activity was determined by the method of TRAP argentation in maligment tumour cells pre- and post-transfected by enhanced vector . Meanwhile the relationship beteewn TK and telomerase was analyzed.Result:①A strong TK gene fluorescent show and TK mRNA expression were displayed after the enhanced suicide gene vector was transfected into nasopharyngeal carcinoma 5-8F cell lines and human breast cancer MCF-7 cell line, which were more stronger than those of mono-promoter group,pGL3-basic-EGFP-TK-hTRETp,and ECV cells transfected by enhanced suicide gene vector. Meanwhile,real-time fluorescent quantified PCR showed that the A value of enhanced vector group was higher than that of controls. ②Telomerase activity after transfection of enhanced vector and GCV was lower than those before by the method of TRAP argentation in nasopharyngeal carcinoma cell lines,but no change in normal control cells after transfection of enhanced vector and GCV.③ After adding GCV, the obvious inhibitory effect of tumour cells growth induced by pGL3-basic-EGFP-TK-hTRETp-CMV enhancer were observed in nasopharyngeal carcinoma 5-8F cell lines and human breast cancer MCF-7 cell line, which was higher than those of mono-promoter, pGL3-basic-EGFP-TK-hTRETp,pGL3-basic-EGFP3 and blank controls, but without inhibitory effect in ECV cells transfected by enhanced vector. Conclusion:TK gene expression is regulated by hTERT promoter and CMV enhancer, and then the telomerase activity is reduced and the cancer cells are specifically killed.But it is unclear how the telomerase are down-regulated by TK gene.
6.Study of change of Th17 cell caused by bacteria biofilm after infection
Guoqiang WANG ; Xiaofang WANG ; Lei WANG ; Huali ZHANG ; Hua ZHONG ; Yajing PAN ; Lei ZHANG ; Tingye LOU
Chinese Journal of Immunology 2015;(1):36-39
Objective:To study of change of Th17 cell and IL-17 caused by bacteria biofilm after infection.Methods: To detect biofilm formation ability of bacteria isolated from bronchus alveolus washing-water of patients;to detect expressing ratios of Th17 cells and IL-17 level in peripheral blood of patients and healthy person;to detect expressing ratios of Th17 cells and IL-17 level in bronchus alveolus washing-water of patients;to analyze the above data using SPSS17.0 software.Results: Expressing ratios of Th17 cells and IL-17 level in peripheral blood of patients infected by producing biofilm and non-producing biofilm bacteria and healthy person are respectively(0.59±0.18)%and(108.8±20.5)pg/ml,(0.58±0.18)%and(100.1±20.7)pg/ml,(0.55±0.17)% and(100.0± 21.4)pg/ml,they were not different among data from patients and healthy person,P>0.05,expressing ratios of Th17 cells and IL-17 level in bronchus alveolus washing-water of patients infected by producing biofilm and non-producing biofilm bacteria were respective (1.37±0.34)%,(157.4±30.8)pg/ml and(1.11±0.21)%,(136.2±24.3)mg/ml,the data between patients infected by producing biofilm and non-producing biofilm bacteria were distinctly different, P<0.05.Conclusion: Bacteria biofilm can cause Th17 cell expressing ratio and IL-17 level to become high in infected part,this may be mechanism that infection become chronic.
7.A novel naphthalene derivative from Aloe barbadensis.
Xiaofang WU ; Jinzhi WAN ; Bingjun LUO ; Miaorong YANG ; Wenjing DING ; Jiasheng ZHONG
Acta Pharmaceutica Sinica 2013;48(5):723-7
To investigate the chemical constituents of A. barbadensis, aqueous extract of the plant was subjected to preparative medium pressure liquid chromatography (MPLC). The chemical structures were mainly determined by spectroscopic evidences (UV, IR, HR-MS, 1H NMR, 13C NMR, HSQC, 1H-1H COSY and HMBC) and chemical methods. A new O, O, O-triglucosylated naphthalene derivative, together with two known 6-phenyl-2-pyrone derivatives and four 5-methylchromones, were isolated and identified as 1-((3-((4- O-beta-D-glucopyranosyl)-beta-D-xylopyranosyloxymethyl)-1-hydroxy-8-alpha-L-rhamnopyranosyloxy)naphthalene-2-y])-ethanone (1), 10-O-beta-D-glucopyranosyl aloenin (2), aloenin B (3), aloesin (4), 8-C-glucosyl-(R)-aloesol (5), 8-C-glucosyl-7-O-methyl-(S)-aloesol (6), and isoaloeresin D (7). Compound 1 is a novel naphthalene derivative and named as aloveroside B, compounds 2-3 are isolated from this Aloe species for the first time.
8.Rapid Identification of Chemical Composition in Aloe Barbadensis Mill by Liquid Chromatography-Mass Spectrometry-Ion Trap-Time-of-Flight
Xiaofang WU ; Jinzhi WAN ; Wenjing DING ; Jiasheng ZHONG ; Yinmao DONG ; Qiaoe WANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(8):1735-1746
This article was aimed to rapidly analyze chemical composition in A loe Barbadensis Mill, and to compare the chemical composition of commercial aloe vera medicinal materials with that of fresh aloe yellow exudate. An opti-mized liquid chromatography-mass spectrometry-ion trap-time-of-flight (LCMS-IT-TOF) method was applied for the analysis of commercial aloe vera medicinal materials and fresh aloe yellow exudates. The Agilent TC-C18 column (4.6 í 250 mm, 5 m) was used. The gradient elution was a solvent system of water(A)-methanol(B). ESI source was operated in both positive and negative ion modes. The results showed that chromones, pyrones, naphthalene deriva-tive, anthrones and anthraquinones were separated successfully, 30 compounds were characterized by the comparison of characteristic MS/MS fragment ions data with the literature. The diagnostic fragmentation patterns of different chemical compositions were also discussed on the basis of EST-IT-TOF MS data. It was concluded that the chemical composition of commercial aloe vera medicinal materials were significantly different from that of fresh aloe yellow ex-udate in terms of types and contents: the former one mainly contains isoaloeresin D and aloin, and few aloesin; but the latter is mainly composed of aloesin and aloin, and the content of aloesin is the highest. The LCMS-IT-TOF analysis can be used to rapidly obtain rich structural information of different chemical compositions, which improves the efficiency of qualitative analysis of chemical composition, and is of great significance to the quality control, eval-uation and the utilization of A loe Barbadensis Mill.
9.Quantitative Analysis for Healing of Fracture of Lower End of Radius by Dual-Energy X-ray Absorptionmetry: A Report of 126 Cases
Qiang FU ; Zhiwei CHEN ; Xunwen CHEN ; Haiwei CAO ; Guangling ZHONG ; Xiaofang LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2001;0(03):-
ive] To explore a new method of evaluating the healing of fractures. [Methods] Self-con trolled trial was applied. Bone mineral density ( BMD) of fracture end (region 3, R 3 ) and its neighboring proximal and distal regions ( R 2 and R 1 respectively) in 126 cases of fracture of lower end of radius was deter mined by dual-energy X-ray absorptionmetry. [Results] BMD of R3 elevated as compared with that of R 2 and R1 (P
10.Relationship between skin prick test reactivity to dermatophagoides farinae and autologous serum reactivity in patients with chronic urticaria
Xiuying HUANG ; Xiaofang ZHOU ; Lin FENG ; Hua ZHONG ; Zhiqiang SONG ; Fei HAO
Chinese Journal of Dermatology 2011;44(1):15-17
Objective To investigate the role of and relationship between reactivity to autologous serum and dermatophagoides farinae (Df) in the pathogenesis of chronic urticaria (CU). Methods Autologous serum skin test (ASST) and skin prick test (SPT) to Df were carried out in 831 patients with CU. The correlation between reactivity to autologous serum and Df was statistically analyzed. Results The positivity rate of ASST and SPT to Df was 51.74% and 64.62%, respectively in the 831 patients. SPT was positive in 56.52% of patients with positive ASST and in 73.86% of those with negative ASST (P < 0.05). In patients with positive ASST, the degree of autologous serum reactivity was negatively correlated with that of reactivity to Df (P < 0.05). Conclusions The skin reactivity to Df and autologous serum plays an important role in the pathogenesis of CU, and the degree of the reactivity to Df and autologous serum is negatively correlated. To conduct ASST and SPT simultaneously in patients with CU may favor the clinical classification and therapy of CU.