0.05),whereas the survival rate in each group of 160 and 320 ?mol/L was significantly higher than that in the control(P

The controversial results of several studies suggest that certain everyday-use chemicals may be linked to breast cancer. ln recent years, extensive researches have been carried out to under-stand breast carcinogenesis and the hedgehog(Hh) signaling pathway has emerged as a critical determi-nant of human breast cancer. Aberrant Hh signaling in adults results in carcinogenesis, angiogenesis, and metastasis. This review is focused on the Hh signaling pathway and chemicals in the regulation of breast cancer development and provide an updated survey of pre-clinical and clinical trials of novel strategies to target them.

Purpose:To set up a new method to screen hMSH2 gene mutations, to detect hMSH2 gene mutations in gynecologic tumor population and to find molecular biomarkers of tumor. Methods:The basic data and blood samples from 42 gynecologic tumor patients werre collected. The genetic mutations of the sixth exon and seventh exon of hMSH2 gene were investigated by multiple polymerase chain reaction single strand conformation polymorphism (PCR SSCP),followed by DNA sequencing. Results:The successes in setting up the multiple PCR SSCP method made it possible to detect hMSH2 gene mutations more quickly and more economically. Two samples of the seventh exon mutation was detected in the tumor population. The mutation is Leu→Phe missense mutation. No mutation of the sixth exon was detected by SSCP.Conclusions:The multiple PCR SSCP method is effective in detecting genetic mutations. There is a mutation site in the seventh exon of hMSH2 gene. It is probably a biomarker of gynecologic tumor. This discovery might offer the basis for further investigation of mutations in large amount population and studies of the function of the mutation.

AIM To explore the molecular mechanism of hydroquinone genotoxicity in human bronchial epithelial cells and investigate whether human X-ray repair cross complementing group 1 (XRCC1)was involved in protecting cells from the damage caused by hydroquinone. METHODS XRCC1 gene was knocked down by RNA interference and XRCC1-deficient cell was established by transfected recombinant plasmid pEGFP-C1-pU6-dsRNA. Normal human bronchial epithelial cells (normal cells) and cells transfected with the empty vector of pEGFP-C1 (vector cells) were used as the normal control and vector control. All cells were treated with different concentrations of hydroquinone (10-100 μmol·L-1) for 4 h. MTT assay was used to test cell viability and comet assay was used to detect the DNA damage and repairment. RESULTS MTT assay showed that hydroquinone inhibited the growth of cells in a concentration-dependant manner and the survival number of XRCC1-deficient cell was less than that of the two control groups. Comet assay revealed that different concentrations of hydroquinone caused more severe DNA damage in XRCC1-deficient cell line than in control cells and there were no significant difference in the two control groups. CONCLUSION The results suggest XRCC1 be involved in preventing cells from damage caused by hydroquinone.

ObjectiveTo evaluate the assistant effct of electroacupuncture on acute incomplete spinal cord injury treated with routine therapy.Methods9 cases who accepted routine therapy, including operation and medicine, were in control group, while other 7 cases who accepted electroacupuncture other than routine therapy were in treating group.ResultsThere were 2 cases were effective, 4 cases were improved and 3 cases were not improved in control group, while 6 cases were clinical recovery, 1 case was effective in treating group.ConclusionElectroacupuncture can improve the effect of routine therapy on acute incomplete spinal cord injury.

目的观察神经外科与针刺联合治疗急性脊髓不完全损伤的疗效。方法神经外科治疗组根据指征确定是否进行手术,药物治疗均给予激素、脱水、抗去甲肾上腺素类药物、神经营养药物,高压氧。神经外科与针刺联合治疗组除上述神经外科治疗外,采用针刺治疗,针刺处方:电针刺激夹脊穴,根据不同辨证配合远端取穴。结果神经外科治疗组9例,持续治疗 3—6个月,出院后随访6个月,6例好转,肌力有所改善,最好者可达到肌力IV—IV+级,3例无效,甚至退步,瘫痪进一步加重。神经外科与针刺联合治疗组病人5例,经针刺及电针治疗10—80天时间后,4例临床治愈,1例经针刺治疗好转后,因转入骨科手术治疗腰椎压缩性骨折时脱离针刺治疗。结论神经外科与针刺联合治疗急性脊髓损伤效果好,并能抑制脊髓损伤的进一步恶化。

OBJECTIVE To explore the effect of bisphenol A (BPA) on the differentiation potential of embryonic stem cells, and provide an experimental basis for evaluation of safety of BPA. METHODS Mouse embryonic fibroblasts (MEFs) and embryonic stem cells (ESCs) were treated with BPA 0.1, 1, 10, 100 and 1000 μmol.L-1 for 8 d respectively. The viability of MEFs and ESCs was measured by CCK-8 and lC50 was calculated. The mRNA expression of α-myosin heavy chain in ESCs was tested by RT-PCR to determine lD50 . The embryonic body cultured by suspension method was treated with BPA 0.001, 0.01, 0.1 and 1 μmol.L-1 for 10 d respectively. The changes of marked genes in each blastoderm were detected by RT-PCR. RESULTS lC50 of BPA to mouse ESCs was 5.22×10-4 mol.L-1 , and to MEFs was 6. 25 × 10-4 mol.L-1 . lD50 of BPA to mouse ESCs differentiating to cardiomyocytes was 7.0×10-7 mol.L-1 . BPA 0.001 and 0.01 μmol.L-1 upregulated the expression of the marked genes of mesoderm, fetal liver kinase-1 and globin transcription factor 1. CONCLUSION BPA is a strong embry-otoxic compound. BPA of low concentration can promote the differentiation of mouse ESCs to mesoderm.

Objective To study the relationship between chemotherapeutic resistance to 5-fluorouracil (5-FU) and Golgi phosphoprotein 3 (GOLPH3) expression in human colon cancer cell line HT29.Methods HT29 cells were divided into four groups:control group,siRNA transfection group,experimental group 1 (30 μmol/L 5-FU),and experimental group 2 (siRNA-GOLPH3 + 30 μmol/L 5-FU).The silencing effect of GOLPH3 gene was detected by RT-PCR and Western blot.The tumor proliferation and formation ability of HT29 cell were measured respectively by MTT and plate colony formation assay.The protein expressions of GOLPH3,P-gp and β-catenin in HT29 cell were detected by Western blot.Results Compared with the control group,the expression level of GOLPH3 mRNA (1.000 ± 0.078 vs.0.147 ± 0.021,t =12.296,P ＜ 0.01) and protein(1.003 ± 0.235 vs.0.077 ± 0.399,t =6.723,P ＜ 0.01),the absorbance(A value) (1.000 ± 0.082 vs.0.769 ± 0.086,t =3.885,P ＜ 0.01) and the tumorigenesis (430 ±36 vs.297 ± 21,t =5.492,P ＜ 0.01) in transfection group significantly reduced.The A value (0.803 ± 0.101 vs.1.050 ± 0.140,t =2.855,P ＜ 0.05;0.242 ± 0.091 vs.1.050 ± 0.140,t =9.664,P ＜ 0.001) and the tumorigenesis (73 ± 8 vs.427 ± 29,t =20.363,P ＜ 0.001;305 ± 22 vs.427 ± 29,t =5.840,P ＜ 0.01) in experimental group 1 and experimental group 2 were significantly lower than that in the control group,while the A value (t =8.264,P ＜ 0.001) and the tumorigenesis (t =17.346,P ＜ 0.001) of experimental group 2 significantly decreased than that experimental group 1.Compared to the control group (0.967 ± 0.094,1.001 ± 0.199,1.000 ± 0.101),the expressions of GOLPH3,P-gp and β-catenin in experimental group 1 (3.634 ± 0.574,2.424 ± 0.261,2.324 ± 0.418) significantly upregulated (t =7.944,P ＜ 0.01;t =7.520,P ＜ 0.01;t =5.330,P ＜ 0.01),while the expressions of these proteins in experimental group 2(0.520 ±0.176,0.466 ±0.159,0.933 ±0.049) were significantly lower than that in experimental group 1 (t =8.983,P ＜ 0.01;t =11.106,P ＜ 0.001;t =5.724,P ＜ 0.01).Conclusion Expression of GOLPH3 is associated with resistance to 5-FU in colon cancer HT29 cell by activating Wnt/β-catenin signal pathway.

Objective: To detect the alterations of telomerase activity and the expression for oxidative stress responsive genes related with senescence during cellular replicative senescence and hydrogen peroxide-induced premature senescence in human embryonic lung fibroblasts (HELFs) in vitro. Methods: The HELFs were divided into young cells (22 population doubling levels, 22PDL) , mid-aged cells (35PDL) and replicative senes-cent cells (49PDL) and premature senescent cells induced by H₂O₂(premature senescence, PS). The telomerase activity was detected by ELISA assay during cellular replicative and premature senescence. The mRNA level of oxidative stress responsive genes related with senescence for Foxo1, Foxo3, Pdx1, apoA-I and MMP1 was per-formed by RT-Q-PCR separately. Results: The mRNA level for Foxo1, Foxo3, apoA-I and Pdx1 was decreased separately during cellular replicative senescence compared to that in the young-stage cells with statistical signifi-cance (P<0.05). The expression of MMP1 was up-regulated 5.1-fold obviously (P<0.05). In premature senes-cence, the mRNA level was only decreased for Foxo1, Foxo3 and apoA-I, but up-regulated 2.3-fold and 6.2-fold for Pdx1 and MMP1 respcetively vs 22PDL significantly (P<0.05). The telomerase activity in young cells was not detected, and it increased in mid-aged cells and replicative senescence stages during cellular replicative se-nescence as compared to 22PDL with statistical significance (P<0.05). The telomerase activity in premature se-nescence was highly active. Conclusion The expression for genes related with senescence has differences be-tween replicative and premature senescence and hydrogen peroxide modifies their expression levels. The telomer-ase activity has been going up with increased PDLs.

OBJECTIVETo put the insight into the trichloroethylene (TCE)-induced effect on the differential expression of subcellular proteins in human normal liver cell line (L-02).

METHODSThe membrane proteins and nuclear proteins of TCE-treated (8.0 mmol/L) group and controls were extracted by subcellular proteome extraction kit, respectively. The TCE-induced differentially expressions were analyzed by a two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization tandem time-of-flight spectrometry (MALDI-TOF-MS). Bioinformatics analysis was used to reveal the biological processes and predict transmembrane domains of differential expressed proteins. The expression of ATP synthase subunit beta (ATP5B), heterogeneous nuclear ribonucleoprotein H2 (hnRNP H2) and far up steam element-binding protein 1 (FUBP1) were measured under TCE treatment by Western blot.

RESULTSAfter TCE treatment for 24 h in L-02 cells, 14 membrane proteins and 18 nuclear proteins were identified as differential expression. After treated with TCE in concentrations of 0, 2.0, 4.0 and 8.0 mmol/L for 24 h, the relative levels of ATP5B expression were 1.00±0.03, 1.21±0.14, 1.25±0.12 and 1.48±0.17 (F = 8.51, P = 0.007), the relative levels of hnRNP H2 expression were 1.00±0.09, 1.22±0.15, 1.43±0.21, 1.53±0.17 (F = 6.57, P = 0.015), respectively; the relative levels of FUBP1 expression were 1.00±0.11, 0.91±0.07, 0.73±0.04 and 0.67±0.03 (F = 15.81, P = 0.001), respectively, which were consistent with the results in proteomics. The bioinformatics analysis showed that the most dominant biological process were involved in RNA processing (10 proteins, P = 2.46×10(-6)), especially in RNA splicing (9 proteins, P = 1.77×10(-7)).

CONCLUSIONThe exposure of TCE could alter the expression of membrane proteins and nuclear proteins in L-02 cells. These abnormal expressed proteins involved in RNA splicing would provide novel clues for further understanding of TCE-induced hepatotoxicity.

Blotting, Western ; Cell Line ; DNA Helicases ; DNA-Binding Proteins ; Hepatocytes ; Heterogeneous-Nuclear Ribonucleoprotein Group F-H ; Humans ; Mitochondrial Proton-Translocating ATPases ; Proteome ; Proteomics ; RNA Processing, Post-Transcriptional ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Trichloroethylene