1.Study on The Effect of Estradiol on Proliferation of Diverse Mammary Primary Cells in Vitro
Zhi LI ; Tao HUANG ; Mingqiang HAN ; Yuan TIAN
Chinese Journal of Bases and Clinics in General Surgery 2003;0(03):-
Objective To understand the effect of estradiol in different concentrations on proliferation of diverse mammary primary cells in vitro. Methods The primary cells of cancer tissue, the adjacent tissue to tumors and normal mammary tissue from patiens with breast cancer were obtained using collagenase digesting method. All the tissue samples were cultivated in vitro, and were given estradiol in different concentrations. The effect of estradiol on the proliferation of those primary cells was measured by MTT. Results Estradiol remarkedly promoted the proliferation of primary cells of cancer tissue and peritumor tissue in vitro, whose ER expression were positive. Whereas, the promotion effect of estradiol on the proliferation of normal mammary primary cells was relatively weak, and there was no correlation between the promotion effect with the expression of ER in cancer tissue. Conclusion The risks of occurrence and relapse of breast cancer would increase significantly when the concentration of estradiol is no less than 103 pmol/L in vivo.
2.In vitro chemosensitivity testing of primary and recurrent breast carcinomas and its clinical significance.
Zhi, LI ; Haiping, SONG ; Wenshan, HE ; Yuan, TIAN ; Tao, HUANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2008;28(6):683-7
In this study, in vitro chemosensitivity testing was conducted on primary cultured breast cancer cells from 96 patients with breast cancer, and the results showed that the cells from a few patients with primary breast cancer developed multidrug resistance (MDR) prior to the first chemotherapy exposure. All the cells from the recurrent cancer patients had MDR. The findings suggested that patients having MDR would benefit from high-dose chemotherapy (HDC) regimens. In vitro chemosensitivity screening, which was aimed at improving the therapeutic efficacy and minimizing side effects, helps in choosing individualized treatment for breast cancer.
Antineoplastic Agents/*pharmacology
;
Breast Neoplasms/*drug therapy
;
Breast Neoplasms/pathology
;
Drug Resistance, Multiple
;
Drug Resistance, Neoplasm
;
Drug Screening Assays, Antitumor/*methods
;
Neoplasm Recurrence, Local/*drug therapy
;
Tumor Cells, Cultured
3.Regulation of Phosphatase and Tensin Homology Deleted on Chromosome 10
tao, XIONG ; de-yuan, LI ; yi, QU ; de-zhi, MU
Journal of Applied Clinical Pediatrics 2004;0(08):-
Phosphatase and tensin homology deleted on chromosome 10 (PTEN) is a tumor suppressor with dual protein and lipid phosphatase activity.PTEN is involved multisystem diseases and cancer,though the mechanisms of PTEN regulation are far from clear.Recent advances concerning regulation of PTEN including protein-protein interaction,phosphorylation,ubiquitination,oxidation and acetylation will be discussed.
4.The chemotherapy role of vinorelbine with relapsed breast cancer primary cells in vitro
Zhi LI ; Chun-Ping LIU ; Peng YUAN ; Yuan TIAN ; Tao HUANG ;
Cancer Research and Clinic 2006;0(08):-
Objective To know the antitumor effect and value of vinorelbine in chemotherapy with relapsed breast cancer.Methods To receive breast cancer tissue from primary and relapsed patients by surgery,then to obtain mammary cancer cells by collagenase.The antitumor effect of 5 chemotherapy drugs with breast cancer cells was measured by culture in vitro and MTT.Results With primary cancer cells from primary breast cancer,the antitumor effect of paclitaxel and docetaxel(sensitivity is 91.04%,92.54% respec- tively) is better than adriamycin,epirubicin and gemcitabine(sensitivity is 73.13%,74.63%,71.64% re- spectively;P
6.Human umbilical cord mesenchymal stem cells may differentiate into Leydig cells through conditioned medium induction.
Xiao-yu XING ; Ji-tao FAN ; Zhi-yuan ZHANG ; Liang ZHONG ; Jie SUN
National Journal of Andrology 2015;21(1):11-16
OBJECTIVETo explore the feasibility of inducing human umbilical cord mesenchymal stem cells (HuMSCs) to differentiate into Leydig cells through conditioned medium derived from Leydig cells.
METHODSHuMSCs and Leydig cells were obtained by tissue blocks culture attachment and enzymatic digestion respectively. HuMSCs were induced by conditioned medium of Leydig cells as an experiment group while those before induction were cultured as a control group. The expressions of LHR, 3β-HSD and StAR in the induced HuMSCs were determined by RT-PCR after 3, 7 and 10 days of culture; those of CYP11A1, CYP17A1 and 3β-HSD measured by immunofluorescence staining after 2 weeks; and that of 3β-HSD detected by Western blot after 4 weeks.
RESULTSThe experimental group showed positively expressed LHR, 3β-HSD and StAR at 3, 7 and 10 days, CYP11A1, CYP17A1 and 3β-HSD at 2 weeks, and 3β-HSD at 4 weeks, while the control group revealed negative expressions at all the time points.
CONCLUSIONInduced with conditioned culture medium derived from Leydig cells, HuMSCs are likely to differentiate into steroidogenic cells and eventually into Leydig cells.
Cell Differentiation ; Culture Media, Conditioned ; Humans ; Leydig Cells ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; Umbilical Cord ; cytology
7.The frequency,phenotypes and invtiro cytotoxic effects of icrculating CD 56+T cells in the patients with chornic HCV infection
Zhaojun DUAN ; Yuhong ZHI ; Lu LONG ; Yuan LIU ; Qiang XU ; Tao SHEN ; Fengmin LU
Chinese Journal of Microbiology and Immunology 2013;(7):481-487
Objective To explore the cell frequency , phenotypes and in vitro cytotoxic effects of circulating CD56+T cells in the patients with chronic HCV infection .Methods Peripheral blood mononu-clear cells (PBMCs) were isolated from 33 patients with HCV chronic infection and 21 healthy subjects. Multi-color flow cytometry was used to analyze cell frequency , expressions of activating receptors ( NKG2C, CD16 and NKp46) and inhibitory receptors (NKG2A and CD158a) on CD56+T cells.The functional mark-er for cytotoxic effects (CD107a) on circulating CD56+T cells and their cytokines expression (IFN-γand TNF-α) with or without stimulation of K 562 human Leukemia cell line were also analyzed .Then the correla-tions among the expressing levels of CD 107 a, IFN-γand TNF-αwere investigated .Results The frequency of CD56+T cells in periphery lymphocytes were significantly decreased in the patients with chronic HCV in -fection as compared with that in healthy controls ( P=0.018 ).The expressions of activating receptors (NKG2C, CD16 and NKp46) on CD56+T cells from HCV infected patients were decreased (P=0.015 for NKG2C, P=0.036 for CD16 and P=0.001 for NKp46), while there was no significant change in the ex-pressions of inhibitory receptors (P>0.05 for both CD158a and NKG2A).The concentrations of IFN-γand TNF-αsecreted by CD56+T cells in the patients with chronic HCV infection were significantly decreased with or without K562 stimulation (P<0.0001).However, in the presence of K562 cells CD107a expression on CD56+T cells were sharply decreased in the patients (P<0.0001).In absence of K562 cells, there was no significant change in CD107a expression on CD56+T cells from patients and healthy controls (P>0.05). The expressions of CD107a, IFN-γand TNF-αwere closely related under the stimulation of K562(r>0.80, P<0.0001).Conclusion The frequency of CD56+T cells was reduced in patients with chronic HCV infec-tion.Moreover, cytotoxic effects and cytokines production mediated by CD 56+T cells were also significantly impaired, indicating that the dysfunction of circulating CD 56+T cells might be associated with the persist-ence of chronic HCV infection .
8.Expression of VEGF, MMPs and its relation with vascular ultrastructure in primary and recurrent gliomas
Zhi-Qiang LI ; Xian-Hou YUAN ; Tao WU ; Zhi-Min WU ; Zhi-Hua WEN
Chinese Journal of Neuromedicine 2005;4(10):994-997
Objective To explore the expression and its implication of angiogenesis and invasiveness related factor in primary and recurrent glioma. Methods Expressions of vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 were detected by immunohistochemical technique. The morphological characteristics of ultrastructure of glioma were observed by transmission electron microscope (TEM). Results The expressions of VEGF, MMP-2 and MMP-9 varied in different grades of primary glioma. With the elevation of the malignant degree of the primary glioma, positive staining rates of VEGF, MMP-2 and MMP-9 increased significantly. The expression of VEGF correlated with both MMP-2 and MMP-9 expression. Compared with the primary glioma, the immunoreactivities of VEGF, MMP-2 and MMP-9 in recurrent glioma increased, especially in those with more severe malignancy. Under transmission electron microscope, endothelial cells markedly proliferated and protruded from the deficiency of basemembrane, concomitantly with edema of the extracapillary gap, plasma extravasation as well as some small worm-eaten caverns in the basemembrane.Conclusion VEGF, MMP-2 and MMP-9 play important roles in glioma angiogenesis and invasiveness.Inhibition of their expressions may be a useful therapy to glioma.
9.Isoflurane induces expression of vascular endothelial growth factor through activating protein kinase C in myocardial cells.
Zhi-gang LIU ; Zhong-yuan XIA ; Xiang-dong CHEN ; Tao LUO
Chinese Journal of Traumatology 2010;13(5):284-288
OBJECTIVEVascular endothelial growth factor (VEGF) plays important roles in establishing collateral circulation of ischemic myocardium. This study aimed to investigate the effect of isoflurane on VEGF expression and the potential intracellular signal transduction pathway in cultured rat myocardial cells in order to further reveal the molecular mechanism of myocardial preservation of isoflurane.
METHODSPrimary myocardial cells of Sprague-Dawley rats were isolated and cultured. They were divided randomly into control group, isoflurane group, protein kinase C (PKC) inhibitor group and PKC inhibitor+isoflurane group where cells were respectively incubated without any treatment, treated by 0.5, 1.0 and 1.5 minimum alveolar concentration (MAC) of isoflurane for 6 hours, by PKC inhibitor calphostin C at a final concentration of 50 nmol/L and by 50 nmol/L calphostin C+1.0 MAC isoflurane for 6 hours. VEGF expression was detected by enzyme-linked immunosorbent assay (ELISA) and the expression levels of PKC isoforms were determined by Western immunoblotting method.
RESULTSIsoflurane increased the VEGF expression in myocardial cells in a dose-dependent way. VEGF levels were significantly higher in 1.0 and 1.5 MAC isoflurane groups than in the control group (both P < 0.01). The effect of isoflurane on upregulating VEGF expression was blocked by PKC inhibitor calphostin C (P < 0.01), but calphostin C did not alter VEGF expression (P > 0.05). Isoflurane induced the activation and translocation of PKCε. Immunoblotting analysis revealed that the immunoreactivity of PKC ε increased significantly in the membrane fractions and deceased significantly in the kytoplasm fractions for cells treated with 1.0 MAC isoflurane as compared with the untreated cells, but not of PKC-α, PKC-δ and PKC-ζ (P less than 0.01).
CONCLUSIONIsoflurane induces myocardial cells to release VEGF through activating PKC-epsilon from the endochylema to the cytomembrane, suggesting a possible novel mechanism of isoflurane protecting myocardial cells.
Anesthetics, Inhalation ; pharmacology ; Animals ; Cells, Cultured ; Dose-Response Relationship, Drug ; Female ; Isoflurane ; pharmacology ; Male ; Myocardial Reperfusion Injury ; prevention & control ; Myocytes, Cardiac ; drug effects ; metabolism ; Protein Kinase C ; antagonists & inhibitors ; physiology ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics
10.Clinical characteristics and prognosis of children aged five or above with stage 3/4 neuroblastoma
Yi ZHANG ; Weiling ZHANG ; Dongsheng HUANG ; Tao HAN ; Tian ZHI ; Jing LI ; Yuan WEN ; Fan LI ; Yanyan MEI ; Yanyan Du
Chinese Journal of General Practitioners 2017;16(1):49-53
Objective To study the clinical characteristics and prognosis of children aged 5 or above with stage 3/4 neuroblastoma ( NB).Method Among 180 children with NB admitted from March 2007 to June 2015, 54 were aged 5 or above with stage 3 or 4.The clinical characteristics , therapeutic efficacy and prognosis of above 54 cases were analyzed.Results There were 36 boys and 18 girls with a male to female ratio of 2∶1.The most common of primary site was retroperitoneum (41/54, 75.5%), followed by mediastinum (10/54,18.5%), spine and pelvic cavity (3/545,6%).After treated by average 9 cycles of chemotherapy , 34 cases ( 63%) achieved complete remission ( CR ) , 13 ( 24.1%) achieved partial remission (PR), 5 (9.3%) presented disease progression (DP), and 2 died (3.7%).Patients were followed up for median 30 months ( 8 -99 months ) , 24 cases died and 30 survived with a overall survival (OS) rate of 55.6%.In 30 survival cases, there were 23 cases (76.7%) of event-free survival (EFS) ,and 6 cases (20.0%) of PR and 1 case (3.3%) of DP.There were significant differences in prognosis among patients with different responses to first therapy (χ2 =8.963, P =0.003 ).Among 20 children with stage 4 NB treated by autologous peripheral blood stem cell transplantation ( APBSCT ) , 13 cases died and 7 survived with an average survival time of (73.55 ±8.89)months.Among 29 cases without APBSCT, 11 cases died and 18 survived with an average survival time of(40.19 ±5.52)months.There was no significant difference in survival between APBSCT and no-APBSCT.Conclusion Children aged 5 or above with NB tend to have advanced stage , relapse and long treatment cycle , but the survival still can be improved with the appropriate treatment .