Objective: To detect the dynamic change of expression of CD133, a marker of liver cancer stem cell, at different stages of liver carcinogenesis induced by chemicals in C57BL/6J mice. Methods: The tumorigenesis and the growth of chemical (diethylnirtosamine/carbon tetrachloride/ethanol)-induced primary HCC (hepatocellular carcinoma) in 50 male C57BL/6J mice were observed. Another 45 male C57BL/6J mice which had not been exposed to carcinogenic chemicals were used as the normal controls. The mice were randomly sacrificed every two weeks, and the liver tissues were removed to observe the change of pathology. The expression of CD133 mRNA was detected by RFQ-PCR (real-time fluorescent quantitative PCR), and the expression of CD133 protein was detected by immunohistochemistry and Western blotting. The percentage of CD133-positive cells was analyzed by FCM (flow cytometry). Results: The primary HCC was successfully induced in male C57BL/6J mice after chemical intervention for 20 weeks. The results of RFQ-PCR and FCM showed that the expression level of CD133 in the chemicalinduced group was obviously higher than that in the normal control group after 8 weeks (P < 0.05, P < 0.001). The expression level of CD133 kept on rising in the chemical-induced group as time progressed, and it was significantly higher at the 16th week than at earlier stages (P < 0.001). Western blotting result showed that the CD133 weak expression could be observed at the 4th week. As time progressed, the expression level of CD133 protein was gradually increased. The result of immunohistochemistry showed that the expressions of CD133 in the chemical-induced group were weakly, moderately and strongly positive at the 8th, 16th and 20th week, respectively; while the expression of CD133 was negative in the normal control group. Conclusion: The liver cancer stem cell marker CD133 is involved in the development and progression of liver cancer, and its expression level is gradually increased in the process of carcinogenesis. Copyright © 2012 by TUMOR.

Adult ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Hygiene ; standards ; Male ; Middle Aged ; Occupational Health ; statistics & numerical data ; Occupational Health Services ; standards ; Young Adult

Career Mobility ; China ; Humans ; Occupational Diseases ; epidemiology ; prevention & control ; Occupational Exposure ; prevention & control ; statistics & numerical data ; Occupational Health

Objective To investigate the expressions of CXCR4 in Barrett esophagus (BE), esophageal adenocarcinoma (EADC) and esophageal squamous cell carcinoma (ESCC), and its relationship with pathology, clinical staging and lymph node metastasis. Methods The expressions of CXCR4 in 56 cases of normal esophageal mucosa, 80 BE (including 22 BE with multifocal dysplasia), 25 EADC and 48 ESCC were examined with immunohistochemical method. Results CXCR4 was expressed in most samples of BE (80. 8% ), EADC (68. 0% ) and ESCC (78.4%) without significant difference ( P ＞ 0. 05 ), which was significantly higher than that in normal esophageal mucosa (39. 3%, P ＜0. 01 ). The level of CXCR4 expression in BE, EADC or ESCC were not related with gender, age, or location of the foci ( P ＞ 0. 05). There was no significant difference in CXCR4 expression between BE without dysplasia or BE with multifocal dysplasia ( P ＞ 0. 05 ). CXCR4 expression level in well-differentiated EADC was significantly higher than that of mild or poorly differentiated (P ＜ 0. 05 ). CXCR4 expression level was higher in EADC with lymph node metastasis than those without ( P ＜ 0. 05 ). CXCR4 level in ESCC with TNM staging grades Ⅲ -Ⅳ was higher than that of grades Ⅰ - Ⅱ, and this variable was also higher in cases with lymph node metastasis than those without (P ＜ 0. 05), so was the case of well and poorly differentiated ESCC (P ＜ 0. 01 ). Conclusion Increased expression level of CXCR4 may be a common feature of EADC and ESCC, which is irrelevant to pathological types. CXCR4 level rises at the stage of BE, which is associated with the degree of tumor differentiation, lymph node metastasis and TNM staging. CXCR4 expression is of guiding significance in the diagnosis of BE, EADC and ESCC, and is the potential drug target.

Objective To develop a visible detection system prepared by protein microarrays which was based on GoldMag particles-labeled technique and compare the results of detection using the protein which was labeled with GoldMag particles and colloidal gold.Methods Human IgG was printed on the glass slides modified with epoxy groups,and goat anti-human IgG conjugated with GoldMag particles and colloidal gold respectlively was then added on the slide.The glass slides were incubated,and then the black images of microarray spots were produced by immuno-gold-silver staining method and observed by naked eyes and recorded with common flatbed scanner.Results The high signal-to-noise ratio could be obtained when the optimized procedures of GoldMag particles-labeling probe were introduced to the protein chip.The conditions of optimum assay were as follows:the spotting concentration of human IgG was 0.2 mg/ml,the glass slides were incubated at 37 ℃ for 2 hour to immobilize human IgG,and the silver enhancement time was 10 min-15 min.Parallelly,the optimized conditions for colloidal gold were as follows:the spotting concentration of human IgG was 0.1 mg/ml,the slides were incubated at 37 ℃ for 1 hour to immobilize human IgG,and the silver enhancement time was 15 min-20 min.Conclusions GoldMag particles-labeled protein technique is comparable to colloidal gold in applying to protein microarrays.The method is considerably simple and practical,and the protein labeled by GoldMag particles could be quantitative.

Aim To study the effect of tubeimosides on human rectal cancer cell line SW480 in vitro and the mechanisms of its antitumor effect.Method The morphological changes were determined by means of light microscopy and electron microscopy respectively.Cell apoptosis was detected by flow cytometry through Annexin V assay and PI fluorescent staining methods.The protein levels of Bc1-2,p53 and Fas were determined using immunohistochemical technique.Results Apparent morphological characteristic of apoptosis was detected under the optical and electric microscope.The percentage of apoptotic cell increased when comparing the drug groups with control after treatment with tubeimoside Ⅰand Ⅲ.Immunohistochemical analysis revealed apoptosis was and induced by tubeimosides through inhibiting Bcl-2 and p53,and upregulating Fas.Conclusion Tubeimosides can induce apoptosis of SW480 cells,which may be one reasons of its antitumor effects.

Cells, Cultured ; Endothelial Cells ; cytology ; Humans ; Stem Cells

Adult ; Female ; Humans ; Logistic Models ; Male ; Mental Health ; statistics & numerical data ; Transients and Migrants ; psychology ; Young Adult

OBJECTIVE:To prepare anti-tumor drug H6(lactone compound)polymeric micelles,and to investigate its in vitro anti-tumor effects. METHODS:Using mPEG2000-PCL4000 as carrier,H6/mPEG2000-PCL4000 micelles were prepared. Using particle size, PDI and 48 h whether to produce precipitation as indexes,feeding ratio,H6 concentration,volume ratio of organic solvent were screened. The encapsulation efficiency and drug-loading amount of micelle were all detected. MTT assay was used to detect the tox-icity of micelles and H6 solution to human non-small cell lung cancer cell A549 and human lung cancer cell H460. RESULTS:The screened formulation was as follows as feeding ratio of 1∶25,H6 concentration of 2 mg/mL,the ratio of ethanol to chloroform of 1∶1(V/V). The parameters of prepared H6/mPEG2000-PCL4000 micelles were as follows as particle size of(40.74±0.116 3)nm,PDI of(0.101±0.006),encapsulation efficiency of(94.87±0.016 3)%,drug-loading amount of(7.07±0.001 5)%(n=3). IC50 of mi-celles and H6 solution to A549 cell were 15.62 and 12.57 nmol/L;IC50 of micelles and H6 solution to H460 cell were 27.68 and 15.19 nmol/L. CONCLUSIONS:H6/mPEG2000-PCL4000 micelles are prepared successfully and show in vitro anti-tumor effects.

ObjectiveTo investigate the effect of glucose and insulin on the synthesis and secretion of adrenomedullin (ADM) by rat aortas in vitro.MethodsThe rat aortas were cut into pieces and divided into several equal groups. All the groups were incubated in K-H buffers including different levels of glucose,insulin and insulin+glucose for 3 hours,the group incubated in K-H buffer without glucose was used as control. To determine ADM in K-H buffers and tissues using RIA method.ResultsADM levels in insulin groups (100.0 μIU/ml,200.0 μIU/ml) and insulin+glucose groups were higher than that in control (P<0.05),the ADM levels in glucose groups and low level insulin group (20.0 μIU/ml) were not significantly difference compared with the control. ConclusionHigh levels of insulin can stimulate the synthesis and secretion of ADM.