Objective To investigate the expression of cyclooxyenase 2 (COX 2)in carcinogensis and development of endometrium carcinoma Methods Immunostainings, westernblotting and quantitve reverse transcription polymerase chain reaction (RT PCR) assay were utilized to measure levels of protein and mRNA expression of COX 2 in following five study groups: 25 cases with proliferative phase, 25 cases with secretory phase,25 cases with endometritis, 23 cases with atypical proliferative phase and 34 cases with endometrium carcinoma Results COX 2 expression of both RNA and protein in patients with endometrial carcinoma was higher significantly than patients with proliferative phase, secretary phase,endometritis,atypical proliferate phase Immunostaining score was 5 46?0 12 vs 3 20?0 18,4 78?0 12,6 10?0 25,8 70?0 93, average absorbent value was 0 75?0 23 vs 0 41?0 45, 0 56?0 31, 1 10?0 56,1 46?0 41;concentration of mRNA [(93?8) vs (65?11),(79?6),(299?11),(493?30) fpg/?g respectively] Successively the expression of COX 2 in atypical proliferation group was higher than normal endometrial and endometritis group The expression in proliferative phase group was higher significantly than secretory phase group ( P

Background and purpose:Previous studies have shown that Bubl was a critical component of the spindle checkpoint.Paclitaxel sensitivity was considered to be dependent on the functionality of this spindle checkpoint.This study investigated the effects of pEGFP-Bubl-shRNA plasmid stably transfected into the cell cycle and its sensitivity in human ovarian cancer cell line A2780.Methods:After the pEGFP-Bubl-shRNA plasmid and empty plasmids were constructed.they were transfected into A2780 cells by the Lipofectamine 2000~(TM).The nontransfected cells were the control.RT-PCR and Western blotting were used to determine the target gene and protein expression.The rate of proliferation inhibition was tested by an MTT assay,apoptosis and cell cycles were determined by flow cytometry,and the mitotic index was determined bv Hoechst33342 dye.Results:RT-PCR and Western blotting showed that pEGFP-Bubl-shRNA/A2780 group displayed a low expression of Bubl compared to the A2780 and pEGFP-Cl/A2780 group(P＜0.05).The sensitivity of the pEGFP-Bubl-shRNA/A2780 group was significantly lower than the non-transfccted and pEGFP-Cl/A2780 cells(P＜0.05).Flow cytometry revealed that the rates of G2/M phase and apoptosis were significantly lower in the pEGFP-Bubl-shRNA/A2780 group than those in the control group (P＜0.05).Conclusion:Bubl plays an important role in the paclitaxel treatment.A down-regulation of Bubl could reduce the drug sensitivity and rate of G_2/M cells in human ovarian cancer cell line A2780.

In order to investigate the roles of MTA2 in the pathogenesis of ovarian epithelial cancer, the expression of MTA2 in 4 ovarian cell lines were detected by semi-quantitative RT-PCR and Western-blot assays. MTA2 expression in normal, borderline, benign and malignant epithelial ovarian tissues was immunohistochemically examined. The expression of MTA2 mRNA and protein was detected in all of 4 cell lines of ovarian epithelial cancer. The expression of MTA2 mRNA and protein was higher in strong migration cell lines than in weak migration ones. In borderline and malignant ovarian tissues tested, MTA2 staining was dramatically stronger than in normal and benign tissues (P < 0.01). The expression levels in malignant ovarian tissues were significantly higher than that in borderline epithelial ovarian tissues (P < 0.01). The expression of MTA2 was correlated with clinical stage, histopathological grade and lymph node metastasis. It was concluded that the high expression of MTA2 was associated with more aggressive behaviors of epithelial ovarian cancer. MTA2 provides a novel indicator of ovarian cancer.

In order to screen the genes differentially expressed in two human prostate cancer cells with different metastasis potentials, suppression subtractive hybridization (SSH) was done twice on human prostate cancer cell line with high potential of metastasis PC3M-1E8 and its synogenetic cell line PC3M-2B4 with low metastasis potential. In the first subtraction PC3M-2B4 was used as tester and PC3M-1E8 as driver and the forward subtractive library was constructed. In the second on the tester and driver were interchanged and the reverse subtractive library was constructed. The screened clones of both libraries were sequenced and Gene Bank homology search was performed. Some clones were confirmed by quantitative real-time PCR. The results showed that two subtractive libraries containing 238 positive clones were constructed. Analysis of 16 sequenced clones randomly picked from two libraries showed that 4 differentially expressed gene fragments were identified as new EST with unknown functions. It was concluded that two subtractive libraries of human prostate cancer cell lines with different metastasis potentials were constructed successfully.
Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Neoplasm Metastasis/*genetics ; Nucleic Acid Hybridization/*methods ; Prostatic Neoplasms/*genetics ; Prostatic Neoplasms/pathology

Objective To investigate the clinical value of echo-tracking(ET) technique in evaluating the carotid elasticity in patients with hyperuricemia and hyperuricemia combined with hyperlipidemia.Methods One hundred and twenty patients with hyperuricemia were divided into two group:group of hyperuricemia (group A) and group of hyperuricemia combined with hyperlipidemia (group B).Sixty normal persons were served as the control group.ET was used to measure the carotid artery elastic modulus including stiffness parameters (β),pressure-strain elastic modulus (Ep),arterial compliance (AC),augmentation index (AI) and pulse wave conducting velocity (PWVβ).Then the statistical data were analyzed to observe the changes of each parameter.Results In the comparison of three groups,elastic function index β,Ep,PWVβ and AI in group A and group B were higher than those of control group (P ＜0.05) ;however,AC was lower than that of control group (P ＜ 0.05).Compared with group A,elastic function index β,Ep,PWVβ,AI was higher in group B,while the AC was lower than that of group B (P ＜0.05).Conclusions ET may be helpful to prevent atherosclerotic changes and to provide the basis for the clinical diagonosis and treatment atherosclerotic changes.

AIM: To investigate the cell cycle arrest ind uced by hypoxia, hypoxia inducible factor-1 and their possible mechanism in huma n ovarian cancer cell line SW626. METHODS: CoCl 2, a chemical inducer of hypoxia and hypoxic cell culture chamber were used to induce chemical and physical hypoxia in human ovar ian cancer cell line SW626. The method of ‘decoy’ was used to block the functi on of HIF-1? because it acts as the core sequence of the target gene as a compe titor combined to the HIF-1?. The cells were divided into group A1 (normal oxyg en), A2 (normal oxygen plus HIF-1? decoy), B1 (CoCl 2), B2 (CoCl 2 plus HIF-1 ? decoy), C1 (hypoxia) and C2 (hypoxia plus HIF-1?). The expression of the HIF -1? protein, mRNA and cell cycle analysis were detected by Western blotting, RT -PCR and flow cytometry (FCM). RESULTS: The expression level of HIF-1? protein in group B1 (3 .75?1.31) and group C1 (3.48?1.01) was significantly higher than that in g roup A1 (0.97?0.31) (P0.05). FCM showed that the G 0/ G 1 phase was markedly increased in group B1 (81.78?24.33) and group C1 (77 .62?22.76) and was significantly higher than that in group A1 (49.49?18.54 ) (P0.05). CONCLUSION: Both CoCl 2 and physical hypoxia could distinctly i nduce cell cycle arrest in G 0/G 1 phase and the expression of HIF-1? in huma n ovarian cancer cell line SW626. HIF-1? plays an important role in cell cycle arrest induced by hypoxia in human ovarian cancer cell line SW626.

Objective To explore the value of virtual touch tissue quantification (VTQ ) in the diagnosis of pancreas early damage of patients with hyperuricemia(HUA) .Methods Sixty‐five cases of patients with normal glucose tolerance and HUA (HUA group) and 42 cases of patients with HUA and impaired glucose tolerance (IGT group) were included in the study ,and other 150 health subjects were taken as control group .After routine ultrasound examination ,the pancreas shear wave velocity values (Vs) of different groups were measured by VTQ technology ,and the results were analyzed .Results Compared with the control group ,the rate of the abnormai ultrasonogram of pancreas in IGT group showed obvious increasing trend but there was no significantly difference between the other groups(both P >0 0.5) .The Vs of the normal control group ,HUA group ,and IGT group were (1 2.1 ± 0 1.5)m/s ,(1 4.1 ± 0 1.3)m/s and (1 5.9 ± 0 1.5)m/s ,respectively .The Vs of HUA group and IGT group were higher than that of the control group (both P <0 0.5) ,and the Vs of IGT group was significantly higher than that of HUA group ( P <0 0.5) .Conclusions Hyperuricemia may cause pancreas damage ,and with the disease progress ,the pancreas damage became more serious ,and VTQ technology may play an important role on the diagnosis of pancreas early damage in HUA patients .

In order to investigate the expression levels of Pin1 mRNA and protein in cervical cancer and its association with Ki67 and their clinical significance, amplification of Pin1 gene was examined by RT-PCR, and the expression of both Pin1 and Ki67 protein was detected by immunohistochemistry in cervical cancer tissues. It was shown that the expression levels of Pin1 were higher in cervical cancer than in normal cervical tissues (P < 0.05). The expression of Pin1 protein was increased progressively along with the disease process from normal cervix to CIN and to cervical cancer (P < 0.05). No significant difference in the Pin1 expression was found between disease stages (FIGO), pathological grades or pelvic lymph node metastasis status (P > 0.05). The expression of Pin1 was significantly higher in adenocarcinoma than in squamous carcinoma of the uterine cervix (P < 0.05). In cervical cancer, the overexpression of Pin1 was positively correlated with that of Ki67 (P < 0.05). These results suggested that the overexpression of Pin1 was closely related with cancer cell proliferation or progression of cervical cancer and contributed to oncogenesis. Pin1 may serve as a potential marker for cervical cancer diagnosis.
Cervical Intraepithelial Neoplasia/metabolism ; Ki-67 Antigen/*biosynthesis ; Ki-67 Antigen/genetics ; Peptidylprolyl Isomerase/*biosynthesis ; Peptidylprolyl Isomerase/genetics ; Tumor Markers, Biological ; Uterine Cervical Neoplasms/*metabolism

Recent evidence has suggested that Akt2 plays an important role in the protection of cells from paclitaxel (PTX)-induced apoptosis and control of the cell cycle. In addition, some scholars suggested that the PTX sensitivity depends on a functional spindle assembly checkpoint. In the present study, we investigated the role of the Akt2/Bub1 cross-talking in apoptosis and cell cycle after exposure of the A2780 ovarian cancer cells to paclitaxel (PTX). Recombinant expression plasmid WT-Akt2 was transfected into A2780 cells by lipofectamine2000, and then the expression level of Akt2 gene was detected by using RT-PCR and Western blotting. Cell apoptosis and cell cycle were detected by flow cytometry and Hoechst 33342 staining after treatment with PTX. Moreover, we compared the expression level of Bub1 in different groups by Western blotting. Our study showed that up-regulation of Akt2 contributed to A2780 ovarian cancer cells overriding PTX-induced G(2)/M arrest, and inhibited Bub1 expression. Our findings might shed light on the molecular mechanism of PTX-induced resistance in ovarian cancer and help develop novel anti-neoplastic strategies.

In this article, the status of spindle assembly checkpoint and the alteration of its major component, Mad2 protein level were examined in A2780 and SKOV3 ovarian cancer cell lines. Recombinant eukaryotic expression plasmid pEGFP-Mad2 was transfected into paclitaxel-resistant SKOV3 cells and Mad2 protein was knocked down by Mad2-specific siRNA in paclitaxel-sensitive A2780 cells. Then the expression level of Mad2 gene was detected by Western blotting. Flow cytometry revealed that SKOV3 cells were not fully arrested in G(2)/M phase in contrast to A2780 cells in the presence of paclitaxel. However, paclitaxel sensitivity assay showed that sensitivity to paclitaxel was reversed after the transfection in both cell lines in terms of number of cells arrested at G(2)/M phase and the expression of Bcl-2 was significantly changed. These results suggest that weakened spindle checkpoint with reduced expression of Mad2 is associated with resistance to paclitaxel in ovarian cells and Bcl-2 may be involved in this process.