The effect of Alternariol Monomethyl Ether (AME), a suspected carcinogen in human being, on the lipid peroxidation in the epithelia of fetal esophagus and stomach was studied. The epithelia of esophagus and stomach of human fetus induced labor with water-bag were treated with AME. Lipid peroxides were measured by thiobarbituric acid test for malondialdehyde (MDA). There was a positive correlation between the values of MDA levels and AME dosaeges. It demonstrated that the lipid peroxdation could be initiated with AME. Lipid peroxidation levels were increased following treatment time elongating. The values of MDA levels in esophagus were higher than those in stomach, which were treated with the same dosaege of AME. It showed that AME had an organ selectivity. These results indicate that AME may be one of genetoxic etiological factors of esophageal carcinoma in Linxian county.

Purpose:To detect DNA polymerase ? gene (pol?) mutations in human gastric cancer specimens and the relationship between H.pylori infection and pol? mutation. Methods:Extracting total RNA from gastric carcinoma,corresponding cancer specimens tissues and normal tissues, synthesizing cDNA and then using them as templates proceed PCR,The products of PCR were checked by SSCP. Extracting DNA from the specimen, we could detect the H.Pylori from the tissue of gastric carcinoma and the tissue adjacent to them by PCR.Results:There were 7 abnormal SSCP of 32 gastric carcinoma samples, and the mutation rate was 21.9%, but nothing abnormal was found in the tissues adjacent to the tumor. The results of H.Pylori DNA were positive in 15 samples from 32 gastric carcinoma tissues. Positive rate was 46.9(15/32).Detection result of tissues adjacent to tumor was consistent with gastric carcinoma. Comparing pol? SSCP to H.pylori-DNA in gastric carcinoma,we found the positive samples of pol? SSCP correlated with H.pylori-DNA.Conclusions:It is suggested that the pol?gene mutation may be associated with carcinogenesis and development of human gastric cancer. H.pylori infection is possibly related to pol? mutation in gastric carcinoma.

Aim To evaluate the value of expressions of CD63 on basophils in diagnosing allergy to penicillins.Methods The expressions of CD63 on basophils activated with 9 kinds of antigens(PG:penicillin G,PV:phenoxomethylpenicillin,AMP:ampicillin,AX:amoxicilloyl,6-APA:6-aminopenicillanic acid,PHA:phenylacetic acid,PHOA:phenoxacetic acid,PHPG:N-(?-hydroxyphenyl)glycine,NPG:N-phenylglycine) were detected by FAST(Flow cytometric allergen stimulation test) in 43 patients with penicillins allergy.Eight types of specific IgE and IgG antibodies to penicillins were determinated by RAST and ELISA,respectively.Results The sensitivity and specificity of CD63 used as a marker in patients with penicillins allergy were 65.12% and 93.33%,respectively.Moreover,the sensitivities of CD63 and specific IgE were higher than those of specific IgG in patients with positive skin test(P

Objective To establish human lung adenocarcinoma multidrug resistance cell lines in vitro,observe their biological characteristics,and investigate the mRNA expressions of DNA pol?,mdr 1,mrp1,GST-?,lrp and topo Ⅱ genes.Methods Paclitaxel-resistant cell lines(A549/TXL20) were established in vitro by exposure to stepwise increased concentrations of the drug in a cell culture medium.Biological morphology and cell cycles were analyzed by morphometry and flow cytometry.The chemoresistance indexes of cells were measured by methyl tetrazolium assay.Evaluation of growth and in vitro drug sensitivity were performed.RT-PCR was employed to analyze the mRNA expressions of the DNA pol?,mdr 1,mrp1,GST-?,lrp,and topo Ⅱ genes.Results ① Compared with parent cells,the resistant sublines had a lower confluent density.They were smaller and mixed with giant cells in different sizes and with different numbers of nucleoli,and the growth property of A549/TXL20 did not change significantly compared with A549 cell lines.② The resistant cells,A549/TXL20,were 19.3 times more resistant to paclitaxel and 67.4 times more resistant to cisplatin than the parent cells,and also demonstrated cross-resistance to mitomycin,vinblastine,and 5-fluouracil(5-FU). ③ Compared with the A549 celllines,an unreasonably higher level of drug resistance and lower drug concentration was detected in A549/TXL20 cells after exposure to the drug in the culture medium.④ The mRNA expression level of DNA pol?,mdr1,GST-?,mrp1 andlrp genes in A549/TXL20 cells was significantly higher than that in A549 cell lines(P

Objective To observe the effect of RNAi that silences MTA1 gene on invasion and migration of esophageal carcinoma 9706 cells. Methods The siRNA expression vector that silences MTA1 gene was transfected into EC9706 cells by liposome. MTA1 mRNA and protein expressions were detected through quantitative RT-PCR and Western blot, respectively. The invasion and migration of EC9706 cells were evaluated by scrape wound healing assay and cell invasion assay in vitro. Results MTA1 gene expression significantly decreased. The scrape wound of EC9706 cells healed more slowly and the cell population that cut through Matrigel were less in the EC9706 cells transfected with siRNA expression vector than non-transfected EC9706 cells and the EC9706 cells transfected with blank vector (P

AIM:To study the effect of p21-activated protein kinase 2 (PAK2) knockdown by RNA interfer-ence on the proliferation and apoptosis of human breast cancer cells .METHODS:The short hairpin RNA ( shRNA) targe-ting PAK2 gene was designed and used for packing lentivirus in 293T cells.Human breast cancer MCF-7 cells were infected by the virus particles and PAK2 knockdown stable cell line was established by puromycin selection .The knockdown effi-ciency was assessed by Western blotting .The proliferation ability of MCF-7 cells was evaluated by CellTiter 96 AQueous and anchorage-independent growth assays .The cell apoptosis induced by staurosporine was detected by flow cytometry . RESULTS:The protein level of PAK2 was significantly suppressed after silencing of PAK2 gene in MCF-7 cells ( P<0.01).Furthermore, knockdown of PAK2 caused remarkable inhibition of the cell proliferation and colony formation (P<0.01).Staurosporine induced more apoptosis in the PAK2 knockdown cells compared with the control cells (P<0.01). CONCLUSION:Knockdown of PAK2 inhibits the proliferation of MCF-7 cells and increases the sensitivity of chemothera-peutic drug-induced cell apoptosis , suggesting that PAK2 might be a new therapeutic target in breast cancer treatment .

Objective To investigate the differentially expressed genes of primary esophageal squamous cell carci-noma and of normal esophageal mucosa. Methods LCM-GMA-cDNA microarray was used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 cases of human esophageal squamous cell carcinoma (ESCC). After high-stringent washing, the cDNA microarray was scanned for the fluores-cent signals. Results Among the 886 target genes, 34 genes had significant difference in Ⅰ / Ⅱ and Ⅲ/Ⅳ group. Cell cycle regulators possibly promoting the growth of tumor cells were highly expressed in the early stages of ESCC, whereas adhesion molecules and extracellular matrix-related molecules possibly promoting invasiveness increased in the later stages. Conclusion More than one gene contributed to esophageal cancer. The profiles of gene expression will bring us chance to understanding the molecular mechanism of tumor progression and to support clinical treat-ment.

AIM: To investigate whether Flk1~+ CD31~- CD34~- cells isolated from human adult adipose tissue have characteristics of hemangioblasts in vivo. METHODS: After sublethally irradiated (300cGy) with a caesium source, the female non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice were injected with human adipose tissue-derived Flk1~+ CD31~- CD34~- cells (10~5 cells per mouse) via tail vain with 0.4 mL Roswell Park Memorial Institute medium (RPMI-1640). The control mice received the same volume of RPMI-1640 medium. All mice were killed 2 months after transplantation for further study. The differentiation potential of Flk1~+ CD31~- CD34~- cells was assessed in bone marrow and gastrointestinal tract by the methods of flow cytometry, RT-PCR, FISH, and triple-color immunofluorescence. RESULTS: Flk1~+ CD31~- CD34~- human adipose tissue-derived adult stem cells differentiated into endothelial cells and hematopoietic cells at the single-cell level in vivo. CONCLUSION: Human adult adipose tissue-derived Flk1~+ CD31~- CD34~- cells bear characteristics of hemangioblast in vivo and may have potential application for the treatment of hematopoietic and vascular diseases.

BACKGROUND: Further studies are needed to understand the cytobiological character, functional regulation, gene changes and expression difference of CD34~+ and CD133~+ stem cells induced by basic fibroblast growth factor (bFGF) using gene chip. OBJECTIVE: To compare the differences of gene expression and the response to bFGF of human umbilical cord CD34~+ and CD133~+ cells, and to explore gene expression changes of bFGF-induced umbilical cord CD34~+ and CD133~+ hematopotic stem cells/hemapoietic progenitor cells in vitro. METHODS: Human umbilical cord blood CD34~+ and CD133~+ cells were isolated and purified by MiniMACS immunomagnetic beads selection. The CD34~+ and CD133~+cells were cultured for 10 to 15 days in DMEM/F12 medium, supplemented with bFGF and B27. Total RNA from these cells was extracted and the genetic level of these cells was performed using Oligo GEArray(r) chip and GEArray software. Selected rate of CD34~+ and CD133~+ hematopoietic stem cells was detected using flow cytometry. CD34~+ and CD133~+ cell morphological changes were measured before and after bFGF induction. The concentration and purity of RNA were determined by agarose gel electrophoresis degeneration. Gene-chip test results were analyzed.

AIM: To construct a mouse IFN-? expression vector and observe the antitumor effects of mouse peritoneal macrophages transfected with IFN-? in vivo and in vitro. METHODS: The IFN-? mRNA was amplified by RT-PCR. The open reading frame of mouse IFN-? gene was recombinanted with eukaryotic expression vector pcDNA3.1 through subcloning. Mouse peritoneal macrophages were transfected with recombinant vector pcDNA3.1-IFN-?. The expression of INF-? mRNA was measured by RT-PCR. Another group of peritoneal macrophages were cultured with the culture medium from pcDNA3.1-IFN-? transfecting groups, and its antitumor effect was measured by MTT. pcDNA3.1-IFN-? plasmid was peritoneally injected inte mouse with tumor. The appearance of ascites of pcDNA3.1-IFN-? plasmid injected mice and survival time were observed. RESULTS: The mouse IFN-? expression vector pcDNA3.1-IFN-? was constructed. The sequence was demonstrated to be the same as on GenBank. The recombinant vector was introduced into mouse peritoneal macrophages. IFN-? mRNA was detected by RT-PCR. The supernatant from cultured macrophages transfected with pcDNA3.1-IFN-? plasmid stimulated the antitumor effects of the macrophages without transfection. The appearance of ascites in pcDNA3.1-IFN-? plasmid injected mouse was delayed and survival time was longer than that in other groups. CONCLUSION: We have successfully constructed the mouse IFN-? expression vector pcDNA3.1-IFN-?. Mouse peritoneal macrophages transfected with pcDNA3.1-IFN-? have antitumor effects in vivo and in vitro.