Objective To investigate the expression and significance of human cervical canceroncogene (HCCR-1) protein in colon carcinoma,colon adenoma and normal colon mucosa.Methods Immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) were used to detect the location and expression of HCCR-1 in the 80 colon adenoma samples,30 colon carcinoma,20 normal colon mucosa.Results The positive expression rate of HCCR-1 protein in normal colorectal mucosa,colorectal adenoma and colorectal carcinoma tissues were 10.00％,62.50％,100.00％,respectively,with significant differences among them(P =0.001).The positive expression rate of HCCR-1 protein in tubular adenoma,mixed pattern of adenoma and villous adenoma were 46.70％,66.70％,80.00％,respectively,with significant differences among them(P =0.015).FQ-PCR showed that the HCCR-1 mRNA expression in normal colorectal mucosa,tubular adenoma,mixed pattem of adenoma,villous adenoma and colorectal carcinoma tissues were 0.609 2 ± 0.326 1,4.034 6 ± 1.188 4,12.353 4 ± 2.828 0,22.563 7 ± 78.952 0,43.202 9 ± 7.675 4.Conclusion HCCR-1 is correlated with canceration of colorectal adenoma and the development of colorectal carcinoma.

Objective To investigate the expression and significance of Ezrin protein in normal colon mucosa, colorectal adenoma and colorectal carcinoma.Methods Eighty colorectal adenoma samples of different pathological types were removed by endoscope in gastroenterology endoscopy center of PLA 252nd Hospital from January 2013 to December 2014, including 30 cases of tubular adenoma, 30 cases of tubulovillous adenoma, and 20 cases of villous adenoma.30 cases of colorectal carcinoma specimens and 30 cases of pericarcinous tissue specimens that were surgically removed in PLA 252nd Hospital were also obtained.The immunohistochemistry was used to detect the expression of Ezrin protein in the sections of tissues, and polymerase chain reaction (PCR) was used to detect the expression level of Ezrin mRNA.Results The positive expression rate of Ezrin protein in normal mucosa group, colorectal adenoma group and colorectal carcinoma group was 6.7%, 52.5% amd 86.7%, respectively with the difference being statistically significant among them (P=0.000).The positive expression rate of Ezrin in tubular adenoma group, tubulovillous adenoma group and villous adenoma group was 33.3%, 60.0% and 70.0% respectively, with the difference being significant among them (P=0.023).The Ezrin mRNA expression in the colorectal carcinoma, villous adenoma, the mixed pattern of adenoma, tubular adenoma and normal colorectal mucosa was 16.466±3.233, 9.778±0.820, 5.966±0.571, 3.043±1.053 and 0.688±0.351 respectively (F=73.628,P=0.017).SNK multiple comparisons displayed that the differences about any two groups of the total mean showed statistical significance.Conclusion The expression of Ezrin may promote proliferation and malignant transformation of colorectal adenoma.

Objective To detect the expression levels of Pokemon in colorectal carcinoma, colorectal adenomas and normal colorectal mucosa, and analyze the relationships among different histological types of colorectal adenomas and colorectal carcinoma.Methods Sixty cases of colorectal carcinoma tissues, 80 cases of colorectal adenomas, and 40 cases of normal colorectal mucosa tissues were collected.Immunohistochemistry and fluorescence quantitative polymerase chain reaction (PCR) were used to detect the expression of Pokemon in the tissues.Results The immunohistochemistry revealed that the positive expression rate of Pokemon protein in normal colorectal mucosa, colorectal adenoma and colorectal carcinoma tissues was 25.0%, 51.2% and 80.0%, respectively, with significant differences among them (P=0.000).The positive expression rate of Pokemon in tubular adenoma, mixed pattern of adenoma and villous adenoma was 33.3%, 53.3% and 75.0%, respectively, also with significant differences among them (P=0.015).There was a significant correlation between the raised expression of Pokemon and unfavorable variables, including nodal metastasis (P=0.040), Dukes stage (P=0.014), and differentiation degree (P=0.004).There was no obvious difference in age (P=0.947) and sex (P=0.795).Fluorescence quantitative PCR showed characterized amplification about Pokemon.The expression levels of Pokemon mRNA in normal colorectal mucosa, tubular adenoma, mixed pattern of adenoma, villous adenoma and colorectal carcinoma were 0.981±0.189, 2.024±0.226, 3.357±0.262, 5.006±0.281, and 6.450±0.216 respectively,F=432.884,P=0.012.Conclusion Pokemon participates in the occurrence and development of colorectal adenoma and colorectal carcinoma.

Objective To explore the expression of minichromosome maintenance protein 2 (MCM2) in normal colorectal mucosa, colorectal adenomas and colorectal carcinoma, and to analyze the relationships among different histological types of colorectal adenomas and colorectal carcinoma.Methods Immunohistochemistry was used to exzamine the location and expression of MCM2 in the colorectal tissues, colorectal adenoma tissues and colorectal carcinoma tissues.Real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) was used to detect MCM2 mRNA and connexin-32 mRNA expression.Results (1) Immunohistochemistry: The positive expression rate of MCM2 protein in normal colorectal mucosa, colorectal adenoma and colorectal carcinoma tissues was 16.7%, 55.0% and 80.0% respectively, with significant differences among them (P=0.031).The positive expression rate of MCM2 in tubular adenoma, mixed pattern of adenoma and villous adenoma was 40.0%, 65.0% and 75.0% respectively, also with significant differences among them (P=0.022).(2) FQ-PCR: The MCM2 mRNA expression in the normal colorectal mucosa, tubular adenoma, the mixed pattern of adenoma, villous adenoma, and colorectal carcinoma was 1.187±0.923, 4.126±1.339, 9.577±0.838, 22.150±4.077 and 48.020±4.811, respectively.The expression of MCM2 gradually increased in colorectal carcinoma, villous adenoma, the mixed pattern of adenoma, tubular adenoma, and normal colorectal mucosa (F=55.512,P=0.029).SNK multiple comparisons displayed that the differences about any two groups of the total mean showed statistical significance.Conclusion MCM2 is correlated with canceration of colorectal adenoma and the development of colorectal carcinoma.Detecting the expression of MCM2 can evaluate the progression of colorectal adenoma and the development of colorectal carcinoma.They may be the molecular biological indicators for predicting canceration of colorectal adenoma.

Objective To investigate the expression and significance of metastasis suppressor 1 (MTSS1) protein in colon adenoma and colon carcinoma.Methods Immunohistochemistry and fluorescence quantitative-polymerase chain reaction (PCR) were used to detect the location and expression of MTSS1 in the 20 normal colon mucosa,30 colon carcinoma,80 colon adenoma samples.Results The positive expression rate of MTSS1 protein in normal colorectal mucosa,colorectal adenoma and colorectal carcinoma tissues were 100.0％,63.8％,6.7％,respectively,with significant differences among them (P =0.005).The positive expression rate of MTSS1 protein in tubular adenoma,mixed pattern of adenoma and villous adenoma were 82.5％,56.7％,35.0％,respectively,with significant differences among them(P =0.005).Fluorescence quantitative PCR showed that the MTSS1 mRNA expression are 36.58 ± 9.06,16.34±1.34,10.47±3.20,3.40±1.28,0.80±0.40.Conclusion MTSS1 is correlated with canceration of colorectal adenoma and the development of colorectal carcinoma.Detecting the expression of MTSSI can evaluate the progression of colorectal adenoma and the development of colorectal carcinoma.

Objective To investigate the expression and significance of Kissl protein in colon adenoma and colon carcinoma.Methods Immunohistochemistry and real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) were used to detect the location and expression of Kiss1 in the 20 normal colon mucosae,30 colon carcinomae,80 colon adenomae.Results The positive expression rate of Kiss1 protein in normal colorectal mucosa,colorectal adenoma and colorectal carcinoma tissues was 93.3％,58.8％ and 5.0％ respectively,with significant differences among them (P =0.005).The posi tive expression rate of Kiss1 protein in tubular adenoma,mixed pattern of adenoma and villous adenoma was 80.0％,66.7％ and 35.0％ respectively,with significant differences among them (P =0.005).FQ-PCR showed that the Kiss1 mRNA expression rate was 28.85 ±8.34,14.61 ± 1.65,7.58 ± 1.31,2.26 ± 1.56 and 0.50 ± 0.40 in normal colorectal mucosa,tubular adenoma,mixed pattern of adenoma,villous adenoma and colorectal carcinoma,respectively.Conclusion Kiss1 is correlated with canceration of colorectal adenoma and the development of colorectal carcinoma.Detecting the expression of Kiss1 can evaluate the progression of colorectal adenoma and the development of colorectal carcinoma.

Objective The aim of this study is to detect the expression levels of Tat interacting protein 30 (TIP30) in colorectal carcinoma,colorectal adenomas,normal colorectal mucosa,and analyze the relationships among different histological types of colorectal adenomas and colorectal carcinoma.Methods We collected 60 colorectal carcinoma tissues,40 normal colorectal mucosa tissues,80 colorectal adenomas.Immunohistochemistry and fluorescence quantitative polymerase chain reaction (PCR) were used to detect the expression of TIP30 in the tissues.Results The results of the immunohistochemistry were that the positive expression rates of TIP30 in normal colorectal mucosa,colorectal adenoma and colorectal carcinoma tissues were 95.0％,67.5％,30.0％,respectively,with significant differences among them (P =0.000).The positive expression rates of TIP30 in tubular adenoma,mixed pattern of adenoma,villous adenoma were 86.7％,66.7％,40.0％,respectively,also with significant differences among them (P =0.003).There was a significant correlation between reduced expression of TIP30 and unfavorable variables,including nodal metastasis (P =0.024),Dukes stage (P =0.008),differentiation degree (P =0.015).There was no obvious difference in age (P =0.815) and sex (P =0.955).Fluorescence quantitative PCR showed characterized amplification about TIP30.The expression of TIP30 mRNA in normal colorectal mucosa,tubular adenoma,mixed pattern of adenoma,villous adenoma and colorectal carcinoma were 25.135 ±1.474,18.701 ±1.596,12.412 ±1.677,6.579 ±0.795,2.137 ±0.535,respectively,F =250.931,P =0.015.Conclusion TIP30 may participate in the inhibition of the occurrence and development of colorectal adenoma and colorectal carcinoma.

Objective To investigate the effects of captlpril on type Ⅰ collagen mRNA expression of culured hepatic stellate cells (HSCs) in vivo Methods HSC-T6 rat HSCs were chosen as the study model of the activated HSCs.Cultured HSCs were randomized into control group,angiotensin Ⅱ (Ang Ⅱ)group (1 × 10-5,1 × 10-7 mol/L) and AngⅡ + captlpril group (1 × 10-5,1 × 10-7 mol/L).HSCs were harvested to detect the collagen Ⅰ mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR).Results Type Ⅰ collagen gene expression levels in different groups of AngⅡ (1 × 10-5,1 × 10-7 mol/L) were 0.850 ± 0.107 and 0.620 ± 0.103 respectively,and that in control group was 0.438 ± 0.061.Type Ⅰ collagen gene expression levels in different groups of Ang Ⅱ + captlpril (1 ×10-5,1 ×10-7 mol/L) were 0.650 ±0.087 and 0.510 ±0.062.Conclusion AngⅡ can increase the type Ⅰ collage mRNA expression of HSCs,which was inhibited by captlpril.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) and captlpril on tissue inhibitorof metalloproteinase-1 (TIMP-1) mRNA expression in culured hepatic stellate cells (HSCs) in vivo.Methods HSC-T6 rat hepatic stellate cell line was chosen as the study model of the activated HSCs.The cultured HSCs were randomized into control group,Ang Ⅱ group (1 × 10-5,1 × 10-7 mol/L) and AngⅡ + captlpril group (1 × 10-5,1 × 10-7 mol/L).HSCs were harvested to measure the TIMP-1 mRNA expression by reverse transcription-polymerase chain reaction (RT-PCR).Results TIMP-1 gene expression levels in different groups of Ang Ⅱ (1 × 10-5 mol/L,1 × 10-7 mol/L) were 1.145 ±0.219 and 0.860±0.115 respectively,and those in control group was 0.523 ± 0.056.TIMP-1 expression levels in different groups of Ang Ⅱ ± captlpril (1 × 10-5 mol/L,1 × 10-7 mol/L) were 0.740 ± 0.089 and 0.590 ± 0.073 respectively,and those in Ang Ⅱ group were 0.850 ± 0.107 and 0.620 ± 0.103 respectively.Conclusion Angiotensin Ⅱ can increase the type Ⅰ collage mRNA expression of HSCs,which can be inhibited by captlpril.

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) and captlpril on proliferation of culured hepatic stellate cells in vivo.Methods HSC-T6 rat hepatic stellate cell line was chosen as the study model of the activated HSCs.Methyl thiazol tetrazolium (MTT) colorimetric assay was used to e valuate the proliferation of HSCs.Results A570 value of control groups was 52.28 ± 1.19 ; A570 values of Ang Ⅱ groups (1 × 10-5 mol/L,1 × 10-7 mol/L,1 × 10-9 mol/L) were 74.81 ±3.61,61.76 ± 1.34,58.12 ± 1.29.A570 values of Captlpril + Ang Ⅱ groups (1 × 10-5 mol/L,1 × 10-7 mol/L,1 × 10-9 mol/L) were 36.32 ±2.11,41.36 ± 1.09,47.01 ±2.65.A570 value of Captlpril + Ang Ⅱ groups was lower than that of control groups (P ＜ 0.01).A570 value of Ang Ⅱ groups was higher than that of control groups (P ＜ 0.01).Conclusion Angiotensin Ⅱ (1 × 10-9 to 1 × 10-5 mmol/L) stimulated proliferation of HSCs as demonstrated by MTT assay.Captlpril + Ang Ⅱ had significantly inhibitory effect on growth of HSCs at the concentration of 1 × 10-9 to 1 × 10-5 mmol/L.

Objective To investigate the effects of rosiglitazone,a ligand of peroxisome proliferators activated receptor gamma (PPAR-γ),on the expression of tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 in hepatic stellate cells (HSCs).Methods HSCs were cultured in medium containing different concentrations of rosiglitazone (5,10,15,20 μmol/L),and harvested at 24 h.The expression of TIMP-1 and TIMP-2 was detected by using reverse transcription polymerase chain reaction.Results TIMP-1 gene expression levels in HSCs cultured with different concentrations of rosiglitazone (5,10,15,20 μmoL/L) were 1.8087 ± 0.0642,1.3517 ± 0.0485,0.9510 ± 0.0835 and 0.5653 ± 0.0879 respectively,and it in control group was 2.0508 ±0.1576.The expression of TIMP-2 in HSCs cultured with different concentrations of rosiglitazone (5,10,15,20 μmoL/L) was 3.1314 ±0.0717,2.0758 ±0.0730,1.4718±0.0623 and 0.8566 ±0.0744 respectively,and it in control group was 3.5784 ±0.0956.Conclusion PPAR-γ ligand rosiglitazone may down-regulate the expression of TIMP-1 and TIMP-2 in HSC dose-dependently to some extent.

Objective To investigate the effects of carthamus tinctorius on gene expression of interstitial collagenase and type Ⅰ collagen in HSC-T6 cells.Methods Hepatic stellate cells (HSC-T) were cultured in medium containing different concentrations of carthamus tinctorius (1.0,and 0.5 g/L) and harvested after incubation for 48 h.Total RNA of HSCs was isolated and gene expression levels of interstitial collagenase (MMP13) and type Ⅰ collagen were detected by using reverse transcription polymerase chain reaction.Results MMP13 gene expression levels in different groups of carthamus tinctorius (1.0,and 0.5 g/L) were 0.63 ±0.10 and 0.40 ±0.07 respectively,and that in control group was 0.26 ±0.04.Type Ⅰ collagen gene expression levds in different groups of carthamus tinctorius (1.0,and 0.5 g/L)were 1.30 ±0.20 and 1.97 ±0.36 respectively,and that in control group was 3.12 ±0.46.Conclusion Carthamus tinctorius may significantly enhance MMP13 gene expression in HSC-T6 ceils,but inhibit type Ⅰ collagen gene expression remarkably in a dose-dependent manner.

Objective To investigate the effects of rosiglitazone,a ligand of peroxisome proliferators activated receptor gamma (PPARγ),on the expression of type Ⅰ collagen (COL Ⅰ) and type Ⅲ collagen (COL Ⅲ) in hepatic stellate cells (HSCs).Methods HSCs were cultured in medium containing different concentrations of rosiglitazone (5,10,15,20 μmol/L respectively),and harvested at 24 h.The expression of COL Ⅰ and COLⅢ was detected by using reverse transcription polymerase chain reaction.Results The COL Ⅰ expression of HSCs cultured with different concentrations of rosiglitazone (5,10,15,20 μmoL/L) was 2.9755 ± 0.0853,1.9866 ± 0.0938,1.4935 ± 0.1078 and 0.9603 ± 0.1365 respectively,and it in control group was 3.4802 ±0.1331.The COLⅢ gene expression in HSCs cultured with different concentrations of rosiglitazone (5,10,15,20 μ mol/L) was 3.6207 ± 0.0461,2.6150 ±0.0793,2.0051 ± 0.0830 and 1.4175 ± 0.0862 respectively,and it in control group was 4.2714 ±0.4652.Conclusion PPARγ ligand rosiglitazone may down-regulate the gene expression of COL Ⅰ and COLll in HSCs dose-dependently to some extent.

Objective To investigate the effects of salvia miltiorrhiza on gene expression of stromelysin-1 (MMP3) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in HSC-T6 cells.Methods HSC-T6 cells were cultured in the medium containing different concentrations of salvia miltiorrhiza (0.8 and 0.4 g/L) and collected after for 48 h.Total RNA of HSC-T6 cells was isolated and gene expression levels of MMP3 and TIMP-2 were measured by using reverse transcription polymerase chain reaction.Results MMP3 gene expression levels in 0.8 and 0.4 g/L salvia miltiorrhiza groups were 2.03 ± 0.32 and 2.19 ±0.26 respectively,and those in control group was 2.13 ±0.30.TIMP-1 gene expression levels in 0.8 and 0.4 g/L salvia miltiorrhiza groups were 2.11 ±0.32 and 3.12 ±0.46 respectively,and those in control group was 4.03 ± 0.56.Conclusion Salvia miltiorrhiza may significantly inhibit TIMP-1 gene expression in HSC-T6 cells in a concentration-dependent manner.Salvia miltiorrhiza has no effect on MMP3 gene expression.