1.Establishment of a nasopharyngeal carcinoma cell line CNE1 stably overexpressing ebv-miR-BART7 mediated by lentivirus.
Cun-cun YUAN ; Hong-bing CAI ; Yu-qi HUANG ; Yan-fen YE ; Man-li ZHAO ; Xiao-ming LÜ ; Xin LI
Journal of Southern Medical University 2011;31(3):419-422
OBJECTIVETo establish a nasopharyngeal carcinoma (NPC) cell line CNE1-pLVTHM/BART7 with stable ebv-miR-BART7 overexpression.
METHODSThe recombinant lentivirus pLVTHM/BART7 expression plasmid was packaged into mature lentivirus by 293FT cells and used to infect CNE1 cells. Flow cytometry was employed for sorting the GFP(+) cells. The efficiency of ebv-miR-BART7 overexpression was determined using qRT-PCR.
RESULTSThe recombinant lentivirus plasmid pLVTHM/BART7 was successfully constructed and verified by PCR and sequencing. The expression of ebv-miR-BART7 in CNE1 cells infected with the lentivirus pLVTHM/BART7 was significantly increased as compared with the negative control and the blank control cells.
CONCLUSIONThe recombinant lentivirus vector pLVTHM/BART7 results in high and stable expression of ebv-miR-BART7 in infected CNE1 cells, which provides a useful cell model for further studies of the role of ebv-miR-BART7 in nasopharyngeal carcinoma.
Carcinoma ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; genetics ; MicroRNAs ; Nasopharyngeal Neoplasms ; genetics ; Plasmids
2.Whole-body fluorescent imaging of the growth and metastasis of GFP-expressing bladder tumors.
Yuan-dong WU ; Wan-long TAN ; Yi XIE ; Zhao-cun YU ; Guo-zhi ZHAO
Journal of Southern Medical University 2006;26(4):436-440
OBJECTIVETo label a human bladder cancer cell line and establish a novel human bladder cancer mouse model.
METHODST-24 cells, a human bladder transitional cell carcinoma cell line, were transfected with GFP plasmid to screen stable GFP-expressing clones. The latter were implanted into the wall of the bladder or the subcutaneous tissue of the neck of nude mice. The growth, invasion, and metastasis of the implanted tumor were observed and evaluated with whole-body optical imaging system. The findings were compared with those of HE staining on routine paraffin sections.
RESULTSGFP-labeled tumor cells displayed green fluorescence under fluorescent microscopy and showed stable GFP expression in vitro and in vivo. One week after in situ transplantation of 5 x 10(5) T24 cells, the new bladder cancer was observed and evaluated under whole-body optical imaging system. Two weeks later, the new bladder tumor could be palpated, and 4 weeks later, metastasis to regional drainage lymph nodes in the pelvic and retroperitoneal lymph nodes occurred. The growth and metastasis of the implant bladder tumor were easily observed and accurately evaluated by fluorescent microscope.
CONCLUSIONGFP-labeled tumor cells display green fluorescence under fluorescent microscopy and show stable GFP expression. GFP-labeled T-24 cells and the novel human bladder cancer model described hereby provide a simple and reliable means for studying human bladder cancer in vivo.
Animals ; Carcinoma, Transitional Cell ; metabolism ; pathology ; Diagnostic Imaging ; Disease Models, Animal ; Female ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Indicators and Reagents ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microscopy, Fluorescence ; Neoplasm Transplantation ; Urinary Bladder Neoplasms ; metabolism ; pathology
3.In vitro inhibition of respiratory syncytial virus with combined ribavirin and an oral preparation of traditional Chinese medicine Hu Fei.
Yu-hua LIU ; An-cun HOU ; Gao-chao ZHAO ; Feng-lian WANG ; Shan-na WU
Chinese Journal of Experimental and Clinical Virology 2003;17(2):187-188
OBJECTIVETo study the in vitro antiviral effect of ribavirin combined with an oral preparation of traditional Chinese medicine "Hu Fei" (protecting the lung) on respiratory syncytial virus (RSV).
METHODSCytopathic effects (CEP) of RSV on Hep2 cells were observed after adding different concentrations of ribavirin, Hu Fei and combination of both into the culture medium.
RESULTSThe minimum concentration of ribavirin and Hu Fei for complete inhibition of CPE caused by RSV was 7.80 microg/ml and 5.00 mg/ml, respectively. When the combination of ribavirin and Hu Fei was applied, their minimum concentrations needed for complete inhibition were decreased to 0.98 ?g/ml and 0.63 mg/ml.
CONCLUSIONSBoth ribavirin and Hu Fei showed in vitro anti-RSV effect, but the inhibitory effect of combined ribavirin and Hu Fei was more potent than either of the preparation alone.
Antiviral Agents ; pharmacology ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Microbial Sensitivity Tests ; Respiratory Syncytial Virus, Human ; drug effects ; Ribavirin ; pharmacology
4.In vitro inhibitory mechanism of respiratory syncytial virus with solution prescription of zhidanhuayu.
Yu-hua LIU ; An-cun HOU ; Gao-chao ZHAO
Chinese Journal of Experimental and Clinical Virology 2008;22(6):486-487
OBJECTIVETo study the effective part of solution prescription of Zhidanhuayu (ZDHY) against respiratory syncytial virus (RSV) in vitro.
METHODSObserve the pathology of RSV to Hep-2 under the condition of different concentrations and each effective part of ZDHY.
RESULTSThe concentration limit causing celluar toxicity of ZDHY is 5.5 mg/ml. The ZDHY failed to block the absorption of RSV to Hep-2 within this concentration, and consequently the cell fell into the full pathological changes. During the concentration of 2.75-5.50 mg/ml, the ZDHY directly destroyed virus array,meanwhile, the infected cells that treated by the medicine kept healthy also.
CONCLUSIONZDHY could not defend the infection of RSV, but is able to destroy the RSV directly and inhibit the RSV inhabiting in the cell.
Animals ; Antiviral Agents ; adverse effects ; therapeutic use ; Cell Line ; Cells, Cultured ; Cercopithecus aethiops ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Respiratory Syncytial Virus Infections ; drug therapy ; Respiratory Syncytial Viruses ; drug effects ; physiology ; Vero Cells ; drug effects ; physiology ; Virus Replication ; drug effects
5.Clinical features of 66 children with acquired immunodeficiency syndrome.
Zai-Cun LI ; Yan ZHAO ; Zhi-Hui DOU ; Lan YU ; Hao WU ; Fu-Jie ZHANG
Chinese Journal of Contemporary Pediatrics 2009;11(2):93-95
OBJECTIVETo study the clinical features of pediatric acquired immunodeficiency syndrome(AIDS).
METHODSThe epidemiological, clinical and laboratory data of 66 children with AIDS were retrospectively studied.
RESULTSOf the 66 patients, 46 (69.7%) were male and 20 (30.3%) were female, with a mean age of 8.7 years (ranged 2-16 years). The mean age at diagnosis was 7.7 years (ranged 2-15 years). Vertical transmission as the route of infection was documented in 48 cases (72.7%). Fourteen children (21.2%) were infected through blood or blood products. The route of infection could not be identified in 4 cases (6.1%). Body weight loss was noted in 43 cases (65.2%), anemia in 42 cases (63.7%), fever in 40 cases (60.6%), fatigue in 38 cases (57.6%), rash in 31 cases (47.0%), chronic cough in 28 cases (12.1%), chronic diarrhea in 24 cases (36.4%), CNS involvement in 16 cases (24.2%), oral thrush in 13 cases (19.7%), and hepatosplenomegaly in 12 cases (18.2%). Body height of 30 cases (45.4%) and body weight of 26 cases (39.4%) ranked the lower level. The immune system was severely suppressed in 59 cases (89.4%) and moderately suppressed in 7 cases (10.6%).
CONCLUSIONSVertical transmission remained the most common route of pediatric HIV infection. There were various clinical manifestations in children with AIDS. The immune systems of the majority of children with this disorder were severely suppressed.
Acquired Immunodeficiency Syndrome ; complications ; etiology ; immunology ; Adolescent ; Body Height ; Child ; Child, Preschool ; Female ; Humans ; Infectious Disease Transmission, Vertical ; Male ; Weight Loss
6.Correlation of vasculogenic mimicry with clinicopathologic features and prognosis of ovarian carcinoma.
Yan GAO ; Xiu-lan ZHAO ; Qiang GU ; Jun-yan WANG ; Shi-wu ZHANG ; Dan-fang ZHANG ; Xing-hui WANG ; Nan ZHAO ; Yu-tong GAO ; Bao-cun SUN
Chinese Journal of Pathology 2009;38(9):585-589
OBJECTIVETo explore the existence of vasculogenic mimicry (VM) in ovarian carcinoma and its correlationship with the clinicopathologic features and prognosis of the tumor.
METHODSA total of 84 ovarian carcinoma cases were collected with complete clinical and prognostic data. CD31 immunohistochemistry and PAS special stain were used to investigate VM in the tumor tissue. Immunohistochemical staining of VEGF, MMP-2, MMP-9, E-cadherin, beta-catenin, and Vimentin were used to explore the pathogenesis of VM.
RESULTSTotally 36 of 84 cases exhibited evidence of VM. FIGO classification, pathologic grades and histological types were significantly different between the VM and non-VM groups. Expression of VEGF, MMP-2, MMP-9, E-cadherin and beta-catenin were higher in the VM group than in the non-VM group. Kaplan-Meier survival curve analysis showed that cases of the VM group had a lower survival rate than that of the non-VM group (P = 0.04).
CONCLUSIONSVasculogenic mimicry exists in ovarian carcinoma. Ovarian carcinomas with a high grade malignancy have a high incidence of VM formation, a higher incidence of metastases and a lower survival rate. High expression of MMP-2 and MMP-9 may contribute to the formation of VM in the ovarian cancer.
Cadherins ; metabolism ; Carcinoma, Endometrioid ; metabolism ; pathology ; Cystadenocarcinoma, Mucinous ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Female ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Neoplasm Metastasis ; Neovascularization, Pathologic ; metabolism ; pathology ; Ovarian Neoplasms ; metabolism ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism ; beta Catenin ; metabolism
7.Expression of TFAR19(PDCD5) in normal human kidney, renal clear cell carcinoma, normal human bladder and bladder carcinoma.
Lin XIONG ; Wan-long TAN ; Zhao-cun YU ; Yuan-dong WU ; He HUANG ; Guo-zhi ZHAO ; Wen-hui ZHU ; Shao-bin ZHENG
Journal of Southern Medical University 2006;26(6):805-809
OBJECTIVETo detect the expression of apoptosis gene PDCD5 in tissues of normal human kidney, renal clear cell carcinoma, normal bladder and bladder carcinoma, and explore the role of PDCD5 gene in renal clear cell carcinoma and bladder carcinoma.
METHODSIndirect immunohistochemistry was employed to detect PDCD5 expression in 63 kidney specimens and 42 bladder specimens. Positive expression rates and intensity of PDCD5 protein expression in the kidney tissue were investigated microscopically and by computerized image analysis. Positive expression rate in the bladder tissue was investigated by microscopic observation.
RESULTSThe results of immunohistochemical staining showed PDCD5 protein overexpression in the renal tubule of normal human kidney tissues and downregulation with the stage increase of renal clear cell carcinoma. PDCD5 protein expression showed statistical significance in tissues of normal kidney and renal clear cell carcinoma in all stages. No obvious PDCD5 expression was detected in the tissues of normal human bladder and bladder carcinoma.
CONCLUSIONPDCD5 is an important apoptosis-regulating factor in the occurrence of renal clear cell carcinoma, and its expression is extremely low in tissues of normal human bladder and bladder carcinoma.
Adult ; Aged ; Apoptosis Regulatory Proteins ; biosynthesis ; Carcinoma, Renal Cell ; metabolism ; Carcinoma, Transitional Cell ; metabolism ; Female ; Humans ; Immunohistochemistry ; Kidney ; metabolism ; Kidney Neoplasms ; metabolism ; Male ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; Urinary Bladder ; metabolism ; Urinary Bladder Neoplasms ; metabolism
8.Effect of Wnt signaling suppression on gefitinib in non small cell lung cancer cell lines.
Xia FANG ; Pan GU ; Cai-cun ZHOU ; Sheng-xiang REN ; Ben-fang LUO ; Yu ZENG ; Yun-jin WU ; Yin-min ZHAO ; Xu-you ZHU ; Xiang-hua YI
Chinese Journal of Pathology 2013;42(7):455-459
OBJECTIVETo explore the effect of Wnt signaling suppression on proliferation of non small cell lung cancer to gefitinib, and its related mechanisms.
METHODSPC9 and PC9/AB2 cells of both gefitinib sensitive and resistant were treated with different concentrations of gefitinib, and the proliferation index was measured using CCK8 kit. The members of Wnt signaling pathway were detected by Western blot. Dual luciferase reportor gene assay (TOP Flash) was used to document the transcriptional level of β-catenin. β-catenin siRNA was transfected into PC9/AB2 cells to suppress the Wnt signaling transcription, followed by treatment with different concentrations of gefitinib. Western blot was then used to detect the expression of EGFR and its downstream signaling after inhibit the expression of β-catenin.
RESULTSTreating with different concentrations of gefitinib, the resistance of PC9/AB2 cells to gefitinib was significantly increased (P < 0.05). The members of Wnt signaling expressed at higher level in PC9/AB2 cells than in PC9 cells (t = 24.590, P = 0.000). TOP Flash examination showed that the endogenous transcriptional activity of Wnt signaling was higher in PC9/AB2 cell than that in PC9 cell (t = 4.983, P = 0.008). Compared with the negative control group, apoptotic rate and sensitivity to gefitinib significantly increased in interfered group (P < 0.05). The expression of p-ERK1/2 significantly decreased after Wnt signaling suppression, although other proteins showed no significant alterations.
CONCLUSIONSuppressing the activity of Wnt signaling can partly reverse the celluar resistance to gefitinib in non small cell lung cancer.
Antineoplastic Agents ; administration & dosage ; pharmacology ; Apoptosis ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Phosphorylation ; Quinazolines ; administration & dosage ; pharmacology ; Wnt Signaling Pathway ; drug effects ; beta Catenin ; metabolism
9.Relationship between programmed cell death 5 protein expression and prognosis of renal clear cell carcinoma.
Wan-long TAN ; Lin XIONG ; Shao-bin ZHENG ; Zhao-cun YU ; Huan QI ; Yue-jun DU ; Peng WU
Journal of Southern Medical University 2006;26(9):1316-1318
OBJECTIVETo study the role of programmed cell death 5 (PDCD5) protein in the oncogenesis and development of renal clear cell carcinoma and its association with the prognosis of the malignancy.
METHODSPDCD5 expression was assayed immunohistochemically in 46 cases of human renal clear cell carcinoma, and the patients' survival was followed up.
RESULTSPDCD5 staining in the adjacent normal tissue of the tumor was significantly stronger than that in the tumor tissue, and PDCD5 expression was significantly correlated with the tumor grade, stage and prognosis. The tumors of high grade with strong invasive ability had much less PDCD5 expression and lighter staining. The three- or five-year survival rates of patients positive for PDCD5 expression was much higher than that of patients negative for PDCD5 expression.
CONCLUSIONPDCD5 is a potent inhibitor of malignant transformation of renal clear cell carcinoma and may serve as a major predictor for evaluating the malignant potential and prognosis of the tumor.
Adult ; Aged ; Apoptosis Regulatory Proteins ; biosynthesis ; Carcinoma, Renal Cell ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; Prognosis ; Retrospective Studies ; Survival Analysis
10.Association of HIV transmissions and non-transmission knowledge with negative attitudes to HIV/AIDS.
Yu-feng WEN ; Hua-dong WANG ; Cun-xi ZHAO ; Ying-shui YAO ; Dong-qing YE ; Zuo-jun JIANG
Chinese Medical Journal 2011;124(4):537-540
BACKGROUNDHuman immunodeficiency virus/acquired immune deficiency syndrome (HIV/AIDS)-related stigma is an obstacle to the implementation of treatment, care, and support programs for people living with HIV/AIDS. This study explored the association of the HIV knowledge with the attitudes toward HIV/AIDS.
METHODSA cross-sectional survey was conducted in HIV epidemic rural areas with 5355 participants. Their knowledge and attitudes about HIV/AIDS were assessed, and the associations of HIV transmission and non-transmission knowledge with negative attitudes towards the HIV/AIDS were analyzed.
RESULTSNegative attitudes were significantly correlated with the HIV non-transmission knowledge and the correlation was higher than that with HIV transmission knowledge among participants who were male, 20 years old and migrant workers, students and respondents of and above junior high school. However, among those who were female, age 30 and older, illiterate with primary school education, negative attitudes were significantly associated with HIV non-transmission knowledge and the association was lower than that with HIV transmission knowledge.
CONCLUSIONSHIV transmission knowledge and non-transmission knowledge have different influences on negative attitudes towards HIV/AIDS among different demographic subgroups.
Acquired Immunodeficiency Syndrome ; transmission ; Adult ; Cross-Sectional Studies ; Female ; HIV Infections ; transmission ; Health Knowledge, Attitudes, Practice ; Humans ; Male ; Middle Aged ; Socioeconomic Factors ; Surveys and Questionnaires ; Young Adult