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1.
Chinese Medical Journal ; (24): 82-87, 2018.
Article in English | WPRIM | ID: wpr-324681

ABSTRACT

<p><b>BACKGROUND</b>Decoy receptor 3 (DcR3) is a protein with anti-apoptotic effect that belongs to the tumor necrosis factor receptor superfamily. DcR3 is highly expressed in a variety of malignant tumors including cholangiocarcinoma and its expression was found to be related to the clinical stage, the invasion, and the metastasis of the tumor. This in vitro study aimed to investigate the effect of downregulated expression of DcR3 on cell viability, cell apoptosis, and cell cycle in cholangiocarcinoma cell line TFK-1.</p><p><b>METHODS</b>Three different cell lines were cultured: human cholangiocarcinoma TFK-1, human biliary epithelial carcinoma HuCCT-1, and human cholangiocarcinoma RBE. The cholangiocarcinoma cell line with the highest expression of DcR3 was selected for further investigation. The expression of DcR3 was silenced/knocked down by transfection with DcR3-siRNA in the selected cell line. Various biological phenotype parameters such as cell viability, apoptosis, and cell cycle were observed.</p><p><b>RESULTS</b>The mRNA and protein levels of DcR3 were measured in the three cell lines, and TFK-1 was selected. After the treatment with DcR3-siRNA for 48 h, DcR3 mRNA and protein expression in the treatment group were 38.45% (P < 0.01) and 48.03% (P < 0.05) of that of the control, respectively. It was found that the cell viability decreased to 61.87% of the control group (P < 0.01) after the downregulation of DcR3 in cholangiocarcinoma cell line TFK-1 by transfection with DcR3-siRNA, while the percentage of apoptotic cells was 2.98 times as compared with the control group (P < 0.05). Compared with the control group the ratio of G0/G1increased, and the ratio of G2/M decreased in the treatment group. However, the differences were not statistically significant.</p><p><b>CONCLUSIONS</b>The effect of DcR3 on the growth and apoptosis of cholangiocarcinoma has been demonstrated. DcR3 is not only a predictive marker for malignant tumor but it is also likely to be a potential target for cancer gene therapy. Further studies should focus on exploring the binding ligand of DcR3, the signaling pathway involved, and the molecular mechanism for the regulation of DcR3 expression in cholangiocarcinoma.</p>

2.
Chinese Medical Journal ; (24): 2320-2331, 2018.
Article in English | WPRIM | ID: wpr-690219

ABSTRACT

<p><b>Background</b>XB130 is a recently discovered adaptor protein that is highly expressed in many malignant tumors, but few studies have investigated its role in hepatocellular carcinoma (HCC). Therefore, this study explored the relationship between this protein and liver cancer and investigated its molecular mechanism of action.</p><p><b>Methods</b>The expression of XB130 between HCC tissues and adjacent nontumor tissues was compared by real-time polymerase chain reaction, immunochemistry, and Western blotting. XB130 silencing was performed using small hairpin RNA. The effect of silencing XB130 was examined using Cell Counting Kit-8, colony assay, wound healing assay, and cell cycle analysis.</p><p><b>Results</b>We found that XB130 was highly expressed in HCC tissues (cancer tissues vs. adjacent tissues: 0.23 ± 0.02 vs. 0.17 ± 0.02, P < 0.05) and liver cancer cell lines, particularly MHCC97H and HepG2 (MHCC97H and HepG2 vs. normal liver cell line LO-2: 2.35 ± 0.26 and 2.04 ± 0.04 vs. 1.00 ± 0.04, respectively, all P < 0.05). The Cell Counting Kit-8 assay, colony formation assay, and xenograft model in nude mice showed that silencing XB130 inhibited cell proliferative ability both in vivo and in vitro, with flow cytometry demonstrating that the cells were arrested in the G0/G1 phase in HepG2 (HepG2 XB130-silenced group [shA] vs. HepG2 scramble group [NA]: 74.32 ± 5.86% vs. 60.21 ± 3.07%, P < 0.05) and that the number of G2/M phase cells was decreased (HepG2 shA vs. HepG2 NA: 8.06 ± 2.41% vs. 18.36 ± 4.42%, P < 0.05). Furthermore, the cell invasion and migration abilities were impaired, and the levels of the epithelial-mesenchymal transition-related indicators vimentin and N-cadherin were decreased, although the level of E-cadherin was increased after silencing XB130. Western blotting showed that the levels of phosphorylated phosphoinositide 3-kinase (PI3K) and phospho-protein kinase B (p-Akt) also increased, although the level of phosphorylated phosphatase and tensin homolog increased, indicating that XB130 activated the PI3K/Akt pathway. Furthermore, we found that a reduction in XB130 increased liver cancer cell sensitivity to tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis.</p><p><b>Conclusions</b>Our findings suggest that XB130 might be used as a predictor of liver cancer as well as one of the targets for its treatment.</p>


Subject(s)
Adaptor Proteins, Signal Transducing , Genetics , Metabolism , Animals , Apoptosis , Carcinoma, Hepatocellular , Metabolism , Pathology , Cell Movement , Cell Proliferation , Gene Knockdown Techniques , Liver Neoplasms , Metabolism , Pathology , Mice , Mice, Nude , Microfilament Proteins , Genetics , Metabolism , Neoplasm Invasiveness , Phosphatidylinositol 3-Kinases , Signal Transduction
3.
Chinese Medical Journal ; (24): 2623-2629, 2016.
Article in English | WPRIM | ID: wpr-230911

ABSTRACT

<p><b>BACKGROUND</b>Decoy receptor 3 (DcR3) binds to Fas ligand (FasL) and inhibits FasL-induced apoptosis. The receptor is overexpressed in hepatocellular carcinoma (HCC), and it is associated with the growth and metastatic spread of tumors. DcR3 holds promises as a new target for the treatment of HCC, but little is known regarding the molecular mechanisms underlying the oncogenic properties of DcR3. The present work, therefore, examined the role of DcR3 in regulating the growth and invasive property of liver cancer cell HepG2.</p><p><b>METHODS</b>HepG2 cells were stably transfected with lentivirus-based short hairpin RNA vector targeting DcR3. After the knockdown of DcR3 was confirmed, cell proliferation, clone formation, ability of migrating across transwell membrane, and wound healing were assessed in vitro. Matrix metalloproteinase-9 (MMP 9) and vascular epithelial growth factor (VEGF)-C and D expressions of the DcR3 knockdown were also studied. Comparisons between multiple groups were done using one-way analysis of variance (ANOVA), while pairwise comparisons were performed using Student's t test. P< 0.05 was regarded statistically significant.</p><p><b>RESULTS</b>DcR3 was overexpressed in HepG2 compared to other HCC cell lines and normal hepatocyte Lo-2. Stable knockdown of DcR3 slowed down the growth of HepG2 (P < 0.05) and reduced the number of clones formed by 50% compared to those without DcR3 knockdown (P < 0.05). The knockdown also reduced the migration of HepG2 across transwell matrix membrane by five folds compared to the control (P < 0.05) and suppressed the closure of scratch wound (P < 0.05). In addition, the messenger RNA levels of MMP 9, VEGF-C, and VEGF-D were significantly suppressed by DcR3 knockdown by 90% when compared with the mock control (P < 0.05).</p><p><b>CONCLUSIONS</b>Loss of DcR3 impaired the growth and invasive property of HCC cell line of HepG2. Targeting DcR3 may be a potential therapeutic approach for the treatment of HCC.</p>


Subject(s)
Analysis of Variance , Cell Movement , Genetics , Physiology , Cell Proliferation , Genetics , Physiology , Hep G2 Cells , Humans , Matrix Metalloproteinase 9 , Genetics , Metabolism , RNA, Small Interfering , Genetics , Receptors, Tumor Necrosis Factor, Member 6b , Genetics , Metabolism , Vascular Endothelial Growth Factor A , Genetics , Metabolism
4.
Article in Chinese | WPRIM | ID: wpr-230417

ABSTRACT

<p><b>OBJECTIVE</b>To compare the clinical effects of two methods of internal fixation in treating unstable femoral intertrochanteric fractures in elderly patients.</p><p><b>METHODS</b>From August 2009 to August 2014, 68 elderly patients with unstable femoral intertrochanteric fracture treated with locking proximal femur plate and auxiliary short reconstructed plate (reconstructing calcar group) and proximal femoral nail antirotation (PFNA group) with clinical course from 1 to 3 days were retrospectively analyzed. In reconstructing calcar group, there were 30 patients including 8 males and 22 females, aged from 63 to 85 years old with an average of (73.41±5.12) years old, the fractures were classified to type AO 31-A2.2 in 12 cases, A2.3 in 11 cases, A3.3 in 7 cases according to AO/ASIF classification. In PFNA group, there were 38 patients including 10 males and 28 females, aged from 65 to 90 years old with an average of (74.26±4.53) years old, the fractures were classified to type AO 31-A2.2 in 15 cases, A2.3 in 13 cases, A3.3 in 10 cases. All fracture were caused by injury, leading pain and swelling. Femoral intertrochanteric fracture was confirmed by X ray films. The data of each group were collected for statistical analysis on the following aspects: the incision length, operation time, blood loss volume, postoperative partial weight bearing standing time, clinical healing time of fracture, postoperative complications, and hip functional score of Harris.</p><p><b>RESULTS</b>All incisions were healed at stage I. In the aspect of postoperative complications, there were 1 case of screw blade cutting and 1 case of deep venous thrombosis in PFNA group; there was 1 case of deep venous thrombosis in the reconstructing calcar group (²=0.000,=1.000). Patients were followed up from 20 to 24 months with an average of 22.5 months. There were no significant in postoperative partial weight bearing standing time, postoperative complications, hip functional score of Harris between two group. There were significant in the incision length, operation time, blood loss volume, clinical healing time of fracture. In the incision length, operation time, blood loss volume, clinical healing time of fracture, the PFNA group was significantly differently less than that of the reconstructing calcar group (<0.001). In the clinical healing time of fracture, the PFNA group was significantly differently less than that of the reconstructing calcar group (<0.05).</p><p><b>CONCLUSIONS</b>For the treatment of unstable femoral intertrochanteric fractures in elderly patients, reconstructing calcar and PFNA are both effective, and proximal femoral intramedullary nails may be the best choice, which can be simpler operation, smaller incision and less healing time.</p>

5.
China Occupational Medicine ; (6): 683-685, 2016.
Article in Chinese | WPRIM | ID: wpr-877007

ABSTRACT

OBJECTIVE: To explore the effect of positive-pressure ventilation on dust removal in whole lung lavage( WLL).METHODS: By random sample method,21 patients with stage Ⅱ or Ⅲ pneumoconiosis were chosen for different WLL.Using the patients' own left and right lung was used for matched control study. The positive pressure ventilation was performed at the end of the 3rd,6th,9th and 12 th lavage in treatment lung( treatment groups). The positive-pressure ventilation was not implemented at the end of the 3rd and 6th lavage in the contralateral lung( control groups) but implemented at the end of the 9th,11 th and 12 th lavage. The recovery of lavage fluid,dust and dust concentration drained from 4th to 9th lavage were compared in the two groups. RESULTS: There was no statistical difference in the recovery of the lavage fluid in the 4th to 9th lavage in the two groups( P > 0. 05). The amount of dust and the dust concentration in the fourth lavage drainage in the treatment group was higher than that in the control group( P < 0. 01). The amount of dust and the dust concentration in the 6th,8th and 9th lavage drainage in the treatment group was lower than that in control group( P < 0. 01). The amount of dust and the dust concentration in the 3rd positive pressure ventilation were higher than that in the 6th positive pressure ventilation in the treatment group( P < 0. 01). The total amount of dust in the treatment group was higher than that in the the control group( P < 0. 01). CONCLUSION: In whole lung lavage,the positive pressure ventilation can accelerate the discharge of dust in the lung of patients with pneumoconiosis.

6.
Chinese Journal of Hepatology ; (12): 822-825, 2010.
Article in Chinese | WPRIM | ID: wpr-360826

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the anti-fibrogenesis property of intraportal vein small interfering RNA (siRNA) injection targeting connective tissue growth factor (CTGF) in a rat model of liver fibrosis induced by carbon tetrachloride (CCl4) and its effect on hepatic stellate cell (HSC) activation.</p><p><b>METHODS</b>24 male rats were randomly divided into four group. rats received CCl4 by subcutaneous injections every three days for 6 consecutive weeks, and meantime they also obtained either siRNA targeting CTGF (as CTGF siRNA group), saline (as model group) or a control siRNA (as control siRNA group) by intraportal vein injection to rats liver at the same approach. Other rats received saline intraportal vein injection for 6 weeks (as normal control group). The expressions of CTGF and a-SMA protein were detected by Western blot. Hepatic histology was evaluated by HE staining and Sirius red staining. The collagen staining areas were measured quantitatively using a computer-aided manipulator with slight modifications. The number of active HSC were evaluated by immunohistochemistry.</p><p><b>RESULTS</b>Six weeks after CCl4 injection, prominent upregulations were observed in the expressions of CTGF and a-SMA protein in saline or control siRNA-treated rats livers. In rats with CTGF siRNA treatment, the protein expressions of CTGF and a-SMA in liver decreased by 95%+/-2% and 86%+/-11% (F=21.234 and 12.473, P<0.01) respectively, the number of active HSC in liver decreased by 76%+/-9% (F=9.179, P<0.01) as compared to the model group. The attenuation of liver fibrosis was also observed in rats with CTGF siRNA treatment.</p><p><b>CONCLUSION</b>Intraportal vein siRNA injection targeting CTGF could significantly inhibit CTGF gene expression in rats, thereby attenuate liver fibrosis by decreasing the number of active HSCs.</p>


Subject(s)
Animals , Connective Tissue Growth Factor , Genetics , Gene Silencing , Hepatic Stellate Cells , Metabolism , Liver Cirrhosis , Genetics , Metabolism , Pathology , Therapeutics , Male , RNA, Small Interfering , Genetics , Rats , Rats, Sprague-Dawley
7.
Chinese Journal of Hepatology ; (12): 840-843, 2008.
Article in Chinese | WPRIM | ID: wpr-250104

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of small interfering RNA targeting connective tissue growth factor (CTGF) on rat transforming growth factor beta (TGF beta)/Smads signal pathway.</p><p><b>METHODS</b>Chemically synthetic siRNA targeting CTGF was transfected into HSC T6 and then they were injected into rat livers through their intraportal veins. At the same time these rats also received CCl4 subcutaneously every three days for 6 consecutive weeks. Untreated HSC T6 or/and rats with random siRNA treatment served as controls. Total RNA or/and protein in HSC T6 and rat hepatic tissues were extracted. The expressions of CTGF and TGF beta 1, Smad2, 3 and 7 genes were detected by reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot.</p><p><b>RESULTS</b>CTGF siRNA significantly reduced the expression of CTGF protein in HSC T6. At 48 h after CTGF siRNA treatment, the down-regulation of CTGF protein was the most significant, up to 94%+/-4% (t=46.196, P less than 0.01), but the expressions of TGF beta 1, Smad2, 3 and 7 mRNA showed no differences in HSC T6 compared with the blank controls. Six weeks after CCl4 injections, prominent up-regulations were observed in the gene expressions of CTGF and TGF beta 1 in saline control or siRNA-treated rat livers. Administering CTGF siRNA for six weeks markedly attenuated the induction of CTGF and TGF beta 1 genes; the expressions of CTGF and TGF beta 1 protein decreased by 95%+/-2% (F=21.234, P less than 0.01) and 74%+/-8% (F=13.464, P less than 0.05), respectively, whereas Smad2, 7 protein expressions were not affected.</p><p><b>CONCLUSION</b>Silencing the CTGF gene can suppress the TGF beta /Smads signal pathway in rat livers.</p>


Subject(s)
Animals , Connective Tissue Growth Factor , Metabolism , Gene Silencing , Male , RNA, Messenger , Genetics , RNA, Small Interfering , Rats , Rats, Sprague-Dawley , Signal Transduction , Smad Proteins , Metabolism , Transfection , Transforming Growth Factor beta , Metabolism
8.
Chinese Journal of Hepatology ; (12): 188-192, 2008.
Article in Chinese | WPRIM | ID: wpr-332286

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the anti-fibrogenesis property of intraportal vein injection of small interfering RNA targeting connective tissue growth factor (CTGF) in a rat model of liver fibrosis and its effect on the accumulation of extracellular matrix (ECM).</p><p><b>METHODS</b>Thirty male rats were randomly divided into five groups. Some rats received CCl4 subcutaneously every three days for 6 consecutive weeks, and in the meantime they also received either siRNA targeting CTGF (preventive group), saline (model group) or siRNA (siRNA control group) by intraportal vein injections. Other rats received CCl4 by subcutaneous injection for 2 weeks, followed by CCl4 and CTGF siRNA intraportal vein injection for 4 more weeks (as treatment group). The expressions of CTGF and type I and III collagen genes were detected by means of reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot respectively. Hepatic histology was evaluated by HE and Sirius red stained sections. The collagen staining areas were measured quantitatively using a computer-aided manipulator with slight modifications. Serum procollagen type III and hyaluronic acid were determined by radioimmunoassay.</p><p><b>RESULTS</b>Six weeks after CCl4 injection, prominent upregulation of gene expressions of CTGF, type I and III collagen, and laminin in saline or siRNA-treated rat livers were observed. The expressions of CTGF at mRNA and protein level and type I and III collagen at mRNA level were markedly reduced in rats with CTGF siRNA treated for four or six weeks. Expressions of CTGF at mRNA and protein levels decreased by 76%+/-8%, 80%+/-3% (F = 68.630) and 95%+/-2%, 93%+/-3% (F = 21.234, P < 0.01); type I and III collagen and laminin at mRNA levels decreased by 74%+/-8%, 78%+/-8%, 31%+/-7% and 57%+/-6%, 59%+/-10%, 43%+/-9% (F = 24.219, 16.315, 9.716, P < 0.01) compared with rats in the model group at 72 h. The CTGF siRNA treatment markedly reduced serum levels of procollagen type III and hyaluronic acid and the degrees of liver fibrosis.</p><p><b>CONCLUSION</b>Intraportal vein siRNA injection targeting CTGF could significantly inhibit CTGF gene expression in rats, thereby attenuating liver fibrosis by reducing ECM accumulation.</p>


Subject(s)
Animals , Carbon Tetrachloride , Connective Tissue Growth Factor , Metabolism , Extracellular Matrix , Metabolism , Gene Silencing , Liver , Metabolism , Pathology , Liver Cirrhosis, Experimental , Metabolism , Pathology , Male , RNA, Small Interfering , Rats , Rats, Sprague-Dawley
9.
Article in Chinese | WPRIM | ID: wpr-253451

ABSTRACT

<p><b>AIM</b>To explore the relationship between the effects of curcumin on cerebral ischemic/reperfusion injury and immediately genic expressions of Fos, Jun and NF-kappaB in hippocampal CA1 area.</p><p><b>METHODS</b>Gerbils were randomly divided into sham group (SH), ischemia/reperfusion group (I/R), curcumin group (CU) and solvent control group (SC). Forebrain ischemia was induced by occlusion of bilateral common carotid arteries. Observations were carried out in each group 15 min, 1 h, 2 h, 6 h, 1 d, 3 d, 5 d and 7 d after ischemia: open field test was used to examine the behavioral change, the apoptosis neurons in hippocampal CA1 region was counted, the expression of Fos, Jun and NF-kappaB in hippocampal CA1 was detected by SABC immunocytochemical technique.</p><p><b>RESULTS</b>The behavioral mark and the number of apoptosis neurons in hippocampal (CA1 region was much less in CU group than in I/R group (P < 0.01) The expression of Fos was more and the expression of Jun and NF-kappaB was less in CA1 area in CU group than in I/R group (P < 0.01).</p><p><b>CONCLUSION</b>Curcumin can significantly protect neurons against cerebral ischemia, increasing the expression Fos and decreasing the expression of Jun and NF-kappaB may be the protective mechanisms.</p>


Subject(s)
Animals , Apoptosis , Brain Ischemia , Metabolism , Pathology , Curcumin , Pharmacology , Gerbillinae , Hippocampus , Metabolism , Male , NF-kappa B , Metabolism , Proto-Oncogene Proteins c-fos , Metabolism , Proto-Oncogene Proteins c-jun , Metabolism , Reperfusion Injury , Metabolism , Pathology
10.
Chinese Journal of Surgery ; (12): 181-185, 2006.
Article in Chinese | WPRIM | ID: wpr-317186

ABSTRACT

<p><b>OBJECTIVE</b>To study lymph node micrometastases (LNMM), expression of nm23-H(1), MMP(9), TIMP(2) proteins, and their relationship and clinical significance in patients with stage Dukes B colorectal cancer.</p><p><b>METHODS</b>Thirty patients with stage Dukes B colorectal cancer were studied. LNMM in these patients was detected by immunohistochemical anti-cytokeratin 20 (CK20) staining. The expression of nm23-H(1), MMP(9) and TIMP(2) proteins in primary tumors was examined by Strept-avidin-biotin complex method. Clinical-pathological data and survival of each patient were recorded and analyzed.</p><p><b>RESULTS</b>(1) The positive dyeing of CK20 was observed in 26.7% for cases and in 7.8% for lymph nodes of 30 patients with stage Dukes B colorectal cancer. (2) Different expression of nm23-H(1) and MMP(9) proteins in the patients between stage Dukes B and stage Dukes CD was observed (P < 0.05). The decreased nm23-H(1) expression, and/or the increased MMP(9) expression in primary stage Dukes B tumors were significantly associated with LNMM (P < 0.05). Sensitivity and specificity for detection of LNMM by using nm23-H(1) or MMP(9) were respectively 62.5% and 81.8% or 75.0% and 69.8%. If by combining nm23-H(1) with MMP(9), specificity for detection of LNMM became 90.9%. The expression of TIMP(2) protein was not related with stage Dukes and LNMM. (3) The percent of tumor recurrence and/or metastasis for the stage Dukes B patients with LNMM was significantly higher than that for the patients without LNMM (P < 0.05), but the survival percent for the patients with LNMM was significantly lower than that for the patients without LNMM. The outcome for the patients with nm23-H(1) (-) LNMM (+) or MMP(9) (+) LNMM (+) was significantly worse than that for patients with nm23-H(1) (+) LNMM (-) or MMP(9) (+) LNMM (-) (P < 0.05).</p><p><b>CONCLUSIONS</b>LNMM is detected by immunohistochemical anti-CK20 staining. The expression of nm23-H(1) and MMP(9) in primary stage Dukes B tumors was significantly associated with LNMM. The outcome in the LNMM patients with nm23-H(1) (-) and/or MMP(9) (+) were worse. Combining examination of CK20 for lymph nodes with expression of nm23-H(1) and MMP(9) for primary tumors is of important clinical significance for staging of Dukes, selection of adjuvant treatment and evaluation of prognosis in patients with colorectal cancer.</p>


Subject(s)
Colorectal Neoplasms , Metabolism , Pathology , Therapeutics , Humans , Keratins , Metabolism , Lymph Nodes , Pathology , Lymphatic Metastasis , Matrix Metalloproteinase 9 , Metabolism , NM23 Nucleoside Diphosphate Kinases , Neoplasm Staging , Nucleoside-Diphosphate Kinase , Metabolism , Prognosis , Tissue Inhibitor of Metalloproteinases , Metabolism
11.
Article in Chinese | WPRIM | ID: wpr-283293

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the clinical significance of detection on lymphatic microvessel, lymphatic microvessel density (LMVD) and vascular endothelial growth factor-C (VEGF-C) in patients with colorectal carcinoma.</p><p><b>METHODS</b>Eighty tissue specimens of the colorectal carcinoma and the peritumoral tissue and thirty of adjacent normal bowel tissue were collected. The lymphatic microvessel and LMVD were determined by 5'-nucleotidase histochemical staining. The expression of VEGF-C protein and VEGF-C mRNA in specimens of colorectal carcinoma and normal colorectal tissues were studied by RT-PCR and immunohistochemical methods utilizing strept-avidin-biotin complex. Clinicopathological data and survival of each patient were obtained and analyzed.</p><p><b>RESULTS</b>(1) The brown or filemot stained lymphatic microvessels were observed in specimens from the colorectal carcinoma, the peritumoral tissue and the normal bowel. Collapsed, nonfunctional lymphatic vessels were observed in the intratumoral tissue, and plenty of lymphatic vessels with large lumen referred as functional lymphatic vessels were observed in the peritumoral tissue. (2) The mean value of LMVD in the peritumoral tissue was significantly higher than that in the normal bowel tissue (9.76+/-2.85 vs. 5.49+/-1.43, t=8.220, P<0.01) and tumor tissue (9.76+/-2.85 vs. 2.13+/-0.96, t=15.118, P<0.001). (3) The positive rate (48.8% vs. 0, P<0.01) and mean value (1.09+/-1.20 vs. 0, P<0.01) of the VEGF-C protein expression in colorectal carcinoma specimens were significantly higher than that of the normal bowel tissue. The expression of VEGF-C protein was consistent with the expression of VEGF-C mRNA. The VEGF-C expression in intratumoral tissue demonstrated significant correlation with LMVD in the peritumoral tissue of colorectal carcinoma. (4) Both LMVD in the peritumoral tissue and the expression of VEGF-C in the intratumoral tissue correlated significantly with Dukes' stage (P<0.0001 and P=0.0234), lymph node metastasis (P<0.0001 and P=0.0059), and survival (P<0.0001 and P<0.0001), but not with age, sex, location and dimension of lesion, gross and histological type. Also, there was a positive significant correlation of LMVD in the peritumoral tissue with degree of differentiation (P=0.0168) and metastasis to the liver or the lung (P=0.0088).</p><p><b>CONCLUSIONS</b>Lymphatic microvessels in the peritumoral tissue are functional. The functional lymphatic microvessels, increased LMVD in the peritumoral tissue and the expression of VEGF-C in the intratumoral tissue may act as the morphological features and the molecular phenotype of lymphangiogenesis in colorectal carcinoma, and also as important predictive markers for evaluating lymphatic metastasis and prognosis in patients with colorectal carcinoma.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Colorectal Neoplasms , Metabolism , Pathology , Female , Humans , Lymphangiogenesis , Lymphatic Metastasis , Lymphatic Vessels , Pathology , Male , Microvessels , Middle Aged , Neoplasm Staging , Prognosis , Vascular Endothelial Growth Factor C , Metabolism , Young Adult
12.
Chinese Journal of Hepatology ; (12): 168-170, 2005.
Article in Chinese | WPRIM | ID: wpr-349178

ABSTRACT

<p><b>OBJECTIVE</b>To study the risk factors of renal failure in the early post-liver transplantation period.</p><p><b>METHODS</b>92 consecutive liver transplantation cases were reviewed and a multi-factor analysis of presumed risk factors of early post-transplantation period renal failure was conducted. The factors analyzed were total bilirubin level, prothrombin activity, onset of structural renal disease, onset of gastrointestinal hemorrhage, whether the patient underwent large-volume paracentesis, or underwent plasmapheresis therapy, needed renal replacement therapy, the operation method used, the bleeding volume during operation and the immunosuppressive agents used.</p><p><b>RESULTS</b>Of the 92 patients, 29 (31.5%) developed acute renal failure (ARF) in the early postoperative period. Multi-factor analysis revealed a high pre-transplantation serum creatinine level and low prothrombin activity as risk factors for development of ARF.</p><p><b>CONCLUSION</b>ARF is a frequent medical complication after liver transplantation. A high pre-transplantation serum creatinine level and low prothrombin activity are risk factors of its development.</p>


Subject(s)
Acute Kidney Injury , Epidemiology , Adolescent , Adult , China , Epidemiology , Creatinine , Blood , Female , Humans , Liver Cirrhosis , General Surgery , Liver Transplantation , Male , Middle Aged , Prothrombin , Metabolism , Risk Factors
13.
Chinese Journal of Hepatology ; (12): 685-688, 2005.
Article in Chinese | WPRIM | ID: wpr-276387

ABSTRACT

<p><b>OBJECTIVE</b>To study the role of caspase-12 expression on hepatocyte apoptosis in an experimental model of acute hepatic failure (AHF).</p><p><b>METHODS</b>A mouse experimental model of AHF was developed by intraperitoneal injection of lipopolysaccharide (LPS) and D-galactosamine (D-Gal). Hepatocyte apoptosis was examined by DNA agarose gel and liver pathology. Caspase-12 mRNA expression in liver was detected by reverse transcriptase PCR (RT-PCR) method. The expression of caspase-12, GRP78 proteins in livers was determined by Western blot.</p><p><b>RESULTS</b>Caspase-12 mRNA expression in the livers increased significantly from 5 to 7 hours after administration of LPS and D-Gal. Typical manifestation of hepatocyte apoptosis appeared at 5 hours after the drug administration. After 5 hours the level of serum ALT and AST were remarkably increased, and they reached the peak at 7 hours. The expression of procaspase-12 protein decreased obviously at 7 hours. Seven hours after the drug administration, hepatocyte apoptosis and necrosis both started. The marker of endoplasmic reticulum (ER) stress, Bip/GRP78 was activated during the development of hepatocyte apoptosis. The level of Bip/GRP78 protein was gradually increased at 5 hours after the drug induction.</p><p><b>CONCLUSION</b>Hepatocyte apoptosis plays an important role in the pathogenesis of AHF. Caspase-12 induced ER stress involves in hepatocyte apoptosis. It suggests that inhibition of caspase-12 activation might be a potential strategy in the treatment of AHF in the future.</p>


Subject(s)
Animals , Apoptosis , Caspase 12 , Genetics , Galactosamine , Hepatocytes , Pathology , Lipopolysaccharides , Liver Failure, Acute , Metabolism , Male , Mice , Mice, Inbred BALB C , RNA, Messenger , Genetics , Random Allocation
14.
Chinese Journal of Hepatology ; (12): 923-926, 2005.
Article in Chinese | WPRIM | ID: wpr-276306

ABSTRACT

<p><b>OBJECTIVES</b>To study the inhibition of primary mouse hepatocyte apoptosis by small interfering RNA (siRNAs) against caspase-12.</p><p><b>METHODS</b>The Balb/c mouse primary hepatocytes were isolated in situ with two-step liver perfusion with 0.5 g/L collagenase type IV, and apoptosis were induced with 4 micromol/L thapsigargin (TG). The three kingds of siRNAs targeting different gene sites (130, 214, 521) were synthetized chemically. The single-stranded RNAs were annealed to produce double-stranded siRNAs, then the mouse primary hepatocytes were transfected by oligofectamine package. The inhibition of caspase-12 was analyzed with RT-PCR and Western-blot. The viable hepatocytes following the induction of apoptosis were evaluated with MTT.</p><p><b>RESULTS</b>All the three kinds of siRNAs could obviously inhibit normal mouse hepatocyte caspase-12 mRNA. The siRNA (214) were more effective than the other two when the concentration was 100 nmol/L. The caspase-12 mRNA expression was inhibited by 52.08%, while that of siRNA (521) was 30.73% (t=4.30, P <0.05). However when the concentration was 200 nmol/L, the inhibitions were similar (88.07%, 86.22% and 89.41% respectively). siRNA (214) could downregulate the expression of apoptotic hepatocytes procaspase-12 by 51.43% ( t=4.30, P <0.01). Contrasted with apoptotic hepatocytes, the cell activity, which was analyzed with MTT, increased by 48.76% (t=2.23, P <0.01).</p><p><b>CONCLUSION</b>siRNAs could effectively downregulate the expression of caspase-12 at mRNA and protein levels and prevent mouse primary hepatocytes from apoptosis.</p>


Subject(s)
Animals , Apoptosis , Physiology , Caspase 12 , Genetics , Hepatocytes , Cell Biology , Male , Mice , Mice, Inbred BALB C , RNA, Messenger , Genetics , RNA, Small Interfering , Genetics
15.
Chinese Journal of Surgery ; (12): 447-449, 2005.
Article in Chinese | WPRIM | ID: wpr-264487

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the variations type of hepatic artery and discuss the method of how to protect hepatic artery from injury during the quick harvest of the donor liver.</p><p><b>METHODS</b>A retrospective review of the variations of hepatic arteries of the donor livers and the course of excision and reconstruction of 200 donor livers was performed, and the aberrance and reconstruction method of hepatic arteries were summarized.</p><p><b>RESULTS</b>37 out of 200 hepatic arteries varied and 2 patients suffered biliary complications because of improper preservation of aberrant hepatic arteries.</p><p><b>CONCLUSIONS</b>Most aberrant liver arteries come from superior mesenteric artery or left gastric artery. Proper quick harvest of multiple organs is the basis of the integrity of hepatic arteries, and all the aberrance must be reconstructed.</p>


Subject(s)
Female , Hepatic Artery , General Surgery , Humans , Kidney Transplantation , Liver Transplantation , Methods , Male , Retrospective Studies , Tissue and Organ Harvesting , Methods , Transplantation, Homologous
16.
Chinese Journal of Hepatology ; (12): 526-529, 2004.
Article in Chinese | WPRIM | ID: wpr-250167

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of chemically synthetic small interfering RNA (siRNA) targeting connective tissue growth factor (CTGF) on the synthesis and secretion of extracellular matrix (ECM) in hepatic stellate cells (HSC).</p><p><b>METHODS</b>Chemically synthetic siRNA targeting CTGF was transfected into HSC T6 (an active HSC line in rats) by oligofectamine package, and untreated HSC T6 were used as control. Total RNA and protein of the cells, after their incubation with siRNA for 24, 48 and 72 hours, were extracted, and the supernatants were collected. The expressions of CTGF and type I and III collagen genes were detected by means of reverse transcription-polymerase chain reaction (RT-PCR) and/or Western blot. Contents of hyaluronic acid and type III pro-collagen in the supernatants were determined by radioimmunoassay.</p><p><b>RESULTS</b>The expression of CTGF at mRNA and protein level and type I and III collagen at mRNA levels were markedly down-regulated in siRNA-transfected HSCs. The contents of hyaluronic acid and type III pro-collagen in the supernatants decreased by 46%+/-7%, 52%+/-7%, 53%+/-7% and 29%+/-18%, 29%+/-7%, 27%+/-5%, compared with those of the blank control at 24, 48 and 72 hours.</p><p><b>CONCLUSIONS</b>Chemically synthetic anti-CTGF siRNA can significantly inhibit CTGF gene expression in HSC, and markedly reduce the synthesis and secretion of ECM including type I and III collagen and hyaluronic acid. The siRNA-directed suppression of CTGF gene in HSC was maintained for 72 hours. This suggests that chemically synthetic siRNA may be a potential in preventing and treating liver fibrosis and may have a promising future for development</p>


Subject(s)
Cell Line , Connective Tissue Growth Factor , Extracellular Matrix , Metabolism , Gene Targeting , Humans , Immediate-Early Proteins , Genetics , Intercellular Signaling Peptides and Proteins , Genetics , Liver , Cell Biology , Metabolism , RNA, Small Interfering , Genetics
17.
Chinese Journal of Surgery ; (12): 1044-1047, 2004.
Article in Chinese | WPRIM | ID: wpr-360905

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the diagnosis and managements of hepatic artery complications in orthotopic liver transplantation.</p><p><b>METHODS</b>The clinical data of 107 consecutive orthotopic liver transplantation patients was reviewed retrospectively to assess the risk factors and the diagnosis and treatment of the vascular complications.</p><p><b>RESULTS</b>The incidence of the artery related complications in orthotopic liver transplantation was associated with the quality of the donor organ artery and the reconstruction way of donor-recipient artery intimately. The main hepatic artery related complications were hepatic artery thrombosis and stenosis. The incidence of the vascular complications was 6.54%, and the mortality rate was 85.7%.</p><p><b>CONCLUSIONS</b>The main influence factors of vascular complications were the quality of the donor organ artery and the reconstruction way of donor-recipient artery. The key steps of organ salvaging and the patients' life saving were early diagnosis and treatment of those complications.</p>


Subject(s)
Adolescent , Adult , Aged , Constriction, Pathologic , Diagnosis , Therapeutics , Female , Hepatic Artery , Pathology , General Surgery , Humans , Liver Transplantation , Male , Middle Aged , Retrospective Studies , Thrombosis , Diagnosis , Therapeutics , Transplantation, Homologous
18.
Chinese Journal of Hepatology ; (12): 298-301, 2003.
Article in Chinese | WPRIM | ID: wpr-344416

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of Tauroursodeoxycholic acid (TUDCA) on Taurodeoxycholic acid (TDCA)-induced HepG2 cell apoptosis and to clarify the molecular mechanism of its anti-apoptosis effect of TUDCA.</p><p><b>METHODS</b>Morphologic evaluation of apoptotic cells was performed by Hoechst 33258 staining and electron microscope. DNA fragment was detected by electrophoresis on 1.5% agarose gels. Apoptosis rate was measured by flow cytometry using PI dye. Following incubation of HepG2 cells either with TDCA alone, or coincubation with TUDCA and TDCA, the releasing level of cytochrome c from mitochondria into cytosol was determined by western blot, also the activity of caspase-3, 8, 9.</p><p><b>RESULTS</b>Incubating the cells with 400 micromol/L TDCA for 12 h induced the cells apoptosis significantly. The apoptotic rate decreased from 50.35% +/- 2.20% to 13.78% +/- 0.84% after coincubation with TUDCA, and this anti-apoptotic effect of TUDCA was confirmed by morphological and DNA ladder detection. TUDCA significantly inhibited the release of cytochrome C from mitochondria into cytosol, and the activity of caspase-9, 3 (t > or = 13.00, P < 0.01), especially at 12 h, caspase-3 activity decreased by 54.9% (t = 16.88, P < 0.01) and 52.5%, however it had no obvious effect on the activity of caspase-8 (t = 1.94, P > 0.05).</p><p><b>CONCLUSIONS</b>TUDCA prevents HepG2 cells apoptosis induced by TDCA through modulating mitochondrial membrane stability, inhibiting the release of cytochrome c and the activation of procaspase-9 and 3. Anti-apoptotic mechanism of TUDCA may be considered to be one of the most important reasons that TUDCA exerts significant efficacy in the treatment of cholestatic liver diseases.</p>


Subject(s)
Apoptosis , Carcinoma, Hepatocellular , Pathology , Caspase 3 , Caspase 9 , Caspases , Metabolism , Cytochromes c , Pharmacology , Humans , Liver Neoplasms , Pathology , Taurochenodeoxycholic Acid , Pharmacology , Taurodeoxycholic Acid , Pharmacology , Tumor Cells, Cultured
19.
Chinese Journal of Surgery ; (12): 10-12, 2003.
Article in Chinese | WPRIM | ID: wpr-257744

ABSTRACT

<p><b>OBJECTIVES</b>To summarize hemodynamic and metabolic changes during bypass, and to evaluate the bypass in liver transplantation.</p><p><b>METHODS</b>Fifty-four patients underwent orthotopic liver transplantation with venovenous bypass from May 2000 to May 2002. Their clinical features were analysed.</p><p><b>RESULTS</b>SHR, MAP, CVP, CO, PaO(2), PaCO(2), serum K(+), Na(+), Ca(2+), BUN values were not significantly changed during bypass. Compared to the pre-bypass stage, pH was decreased in the post-bypass stage (P < 0.05), serum lactic acid value was increased in the bypass and post-bypass stage (P < 0.05), active clotting time was increased in the bypass stage (P < 0.05), serum creatinine value was increased on first postoperative day (P < 0.05).</p><p><b>CONCLUSIONS</b>Venovenous bypass could improve hemodynamic and metabolic stability in the anhepatic phase, but it also could increase operation duration, liver ischemic time and cost.</p>


Subject(s)
Adolescent , Adult , Aged , Female , Hemodynamics , Humans , Kidney , Liver Transplantation , Methods , Male , Middle Aged , Portal Vein , General Surgery , Postoperative Complications
20.
Article in Chinese | WPRIM | ID: wpr-344258

ABSTRACT

The treatment planning can directly decide the treatment effect of X-knife. This paper presents some qualitative and quantitative tools assessing the treatment planning, mainly they are the isodose curve, dose-wash, volume dose and dose volume histogram.


Subject(s)
Algorithms , Brain Neoplasms , General Surgery , Evaluation Studies as Topic , Humans , Radiosurgery , Radiotherapy Dosage , Radiotherapy Planning, Computer-Assisted , Methods , Radiotherapy, Conformal , Methods , Sensitivity and Specificity
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