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Objective:To explore the medium-long term efficacy of transjugular intrahepatic portosystemic shunt (TIPS) for gastrointestinal hemorrhage in patients with idiopathic non-cirrhotic portal hypertension (INCPH).Methods:From March 2013 to July 2018, clinical data of 13 INCPH patients, including 5 males, 8 females,with gastrointestinal hemorrhage were retrospectively analyzed, who were diagnosed at the First Affiliated Hospital of Zhengzhou University, Anyang Fifth People′ s Hospital and Yuncheng Central Hospital. All patients received TIPS treatment. The general information, postoperative survival rate, the incidence of rebleeding, shunt dysfunction rate, and incidence of hepatic encephalopathy were analyzed.Results:All 13 patients with INCPH completed TIPS successfully with an average age of 45±8 (33 to 59) years. The hepatic venous pressure gradient (HVPG) decreased from 20.0-26.0 (22.6±1.9) mmHg before procedure to 8.0-14.0 (9.4±3.2) mmHg after. The median follow-up time was 44±7 (31 to 53) months. One patient died of liver failure 27 months after TIPS. Hepatic encephalopathy occurred cumulatively in 1 case (1/13), 1 case (1/13) and 1 case (1/13) in 12, 24 and 36 months after TIPS. Stent restenosis occurred cumulatively in 2 cases (2/13), 3 cases (3/13) and 3 cases (3/13) in 12, 24 and 36 months after TIPS. Portal vein thrombosis occurred cumulatively in 2 cases (2/13), and no primary liver cancer developed.Conclusions:TIPS is safe and effective in the treatment of INCPH with gastrointestinal bleeding with favorable medium-long term outcome.
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Objective:To investigate the clinical value of transjugular liver biopsy (TJLB) in patients with unexplained liver disease complicated with massive ascites or coagulopathy.Methods:A retrospective analysis was performed from patients underwent TJLB in the First Affiliated Hospital of Zhengzhou University, Zhoukou Central Hospital, Shangqiu First People's Hospital and Jincheng People's Hospital from March 2015 to January 2022 due to unexplained liver disease complicated with massive ascites or coagulopathy. A total of 37 patients were included, including 21 males and 16 females, aged (53.5±11.9) years. According to different puncture points, the patients were divided into two groups: transhepatic right vein TJBL and transhepatic middle vein TJBL. The obtained liver tissue sampling effect, puncture times, complications were analyzed.Results:The success rate of TJLB was 97.3%(36/37). Thirty-six patients were able to obtain more than three segments of liver tissue and obtain histological diagnosis, and the pathological diagnosis rate was 100.0%(36/36). The number of puncture times, the amount of hepatic tissue and the number of portal areas in the right hepatic vein group (21 cases) were (3.7±0.9), (3.7±0.7) and (6.5±0.9) respectively, and those in the middle hepatic vein group (15 cases) were (3.7±0.7), (3.7±0.7) and (6.3±0.8) respectively. There were no significant differences between the two groups (all P>0.05). Conclusion:TJLB is safe and feasible for patients with unexplained liver disease complicated with massive peritoneal effusion and coagulopathy. Good liver tissue specimens can be obtained by TJLB from both right hepatic vein and middle hepatic vein.
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Objective To design a new integrated portable biliary internal-external drainage catheter carrying125 I seeds used for the treatment of malignant biliary obstruction lesions so as to achieve the dual curative effects of biliary drainage and brachytherapy. Methods A total of 15 patients with malignant obstructive jaundice, who were admitted to the First Affiliated Hospital of Zhengzhou University, China, during the period from September 2016 to January 2018, were enrolled in this study. Biliary stent implantation was performed in all patients, which was followed by insertion of a new integrated portable biliary internal-external drainage catheter carrying125 I seeds. The technical success rate, clinical success rate, complications, stent patency time and patient survival rate were evaluated. Results The placement of the drainage tube was simple and smooth, and the technical procedure was successful in all patients. One month after treatment, the bilirubin level was decreased significantly when compared with preoperative one (P<0.01), while the blood indexes and immunological indicators showed no obvious changes (P>0.05) . After treatment, 2 patients (13.3％) developed cholangitis and 2 patients (13.3％) had small amount of biliary bleeding, which returned to normal after symptomatic treatment. No severe complications such as perforation of bile duct, massive bleeding, radiation enteritis and radioactive source leakage, etc., occurred. The patients were followed up for 55-402 days, 6 patients (40.0％) developed biliary re-obstruction. The median patency time of stent was 255 days, and 6-month stent patency rate was 64.5％. Five patients died and 10 patients survived, the 9-month survival rate was 64.3％, the median survival time was 368 days. Conclusion By using the new integrated portable biliary internal-external drainage catheter carrying125 I seeds, the effects of bile drainage and brachytherapy can be simultaneously achieved. Preliminary clinical practice indicates that this new drainage catheter is feasible, safe and effective, although its long-term efficacy needs to be clarified with further follow-up observations and controlled studies.
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Objective:To investigate the expression of transmembrane 4 super family 1 (TM4SF1)in breast cancer tissue,and to elucidate its clinical significance and explore the related molecular biological mechanisms. Methods:A total of 190 cases of human breast cancer,110 cases of paracancerous tissue and 110 cases of normal breast tissue were collected.Immunohistochemistry was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue,paracancerous tissue,and normal breast tissue;Western blotting method was used to detect the expression levels of TM4SF1 in breast cancer tissue,paracancerous tissue,and normal breast tissue;RT-PCR method was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue,paracancerous tissue, and normal breast tissue.The positive expression rates of TM4SF1 in breast cancer tissue of the breast cancer patients with different clinicopathological features were detected.Results:The positive expression rate of TM4SF1 in the breast cancer tissue was significantly higher than those in paracancerous tissue and normal breast tissue (P <0.05);there was no significant difference in the positive expression rates of TM4SF1 between paracancerous tissue and normal breast tissue (P = 0.531);the expression of TM4SF1 was not correlated with age,but was closely correlated with tumor size,differentiation degree,lymph node metastasis and tumor stage (P <0.05);the positive expression rate of TM4SF1 in basal like breast cancer tissue was higher than those in the other three types of tissues (P <0.05).The results of Western blotting showed that the expression level of TM4SF1 in breast cancer tissue was higher than those in paracancerous tissue and normal breast tissue (P < 0.05 ), but there was no significant difference in the expression level of TM4SF1 between paracancerous tissue and normal breast tissue (P >0.05). The results of RT-PCR showed that the expression level of TM4SF1 mRNA in breast cancer tissue was higher than those in the paracancerous tissue and normal breast tissue (P <0.01);there was no significant difference in the expression level of TM4SF1 mRNA between paracancerous tissue and normal breast tissue (P > 0.05 ). Conclusion:The TM4SF1 is highly expressed in breast cancer tissue. TM4SF1 may affect the occurrence, development and distant metastasis of breast cancer through various mechanisms.TM4SF1 may be a potential target for the treatment of breast cancer.
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Objective To establish the A549 cell line with stable expression of HIF1α by using lentiviral vector system.Methods Primers were designed and synthesized with human HIF1α gene coding sequence by the National Center of Biotechnogical Information(NCBI) as the template.HIF1α was amplified by PCR.The HIF1α fragment recycled by enzyme digestion was recombined with prepared lentiviral vector HBLV-RFP-Puro.The recombinant plasmid was identified by PCR and gene sequencing.The recombinant plasmid and the auxiliary plasmid were co-transfect into 293T cell.After filtration and concentration of packaged virus,the viral titer was detected by using the dilution counting method.The prepared lentivirus was infected A549 cells.The drug screening was adopted to stabilize the transfected cell line.The transfection effect was detected and observed by fluorescence microscope and Western blotting.Results The HIF1α fragment amplified by PCR was successfully verified and the recombinant plasmid was successfully constructed by PCR and gene sequencing identification.High-titer LV-HIF1α was obtained by successful package.After LV-HIF1α infecting A549 cells,the cells showed the red fluorescence by fluorescence microscope.The expression level of HIF1α in the LV-HIF1α group was significant higher than that in the control group by Western blot.Conclusion The 549 cell line with HIF1α stable expression mediated by lentivirus is constructed successfully.
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Objective to construct the high titers rat Hes1 adenovirus expression vector (Ad-Hes1).Methods With the rat cDNA as a template,the Hes1 fragment was amplified by PCR,which constructed pShuttle-CMV-Hes1 shuttle plasmid by directly clone.Based on pShuttle-CMV-Hes1,pAdeno-Hes1 virus plasmid was constructed,pAdeno-Hes1 was transfected into 293 cells to package Ad-Hes1,virus titers were determined by modified TCID50.Hes1 was detected by Western blot after Ad-Hes1 infected with H9c2 myocardial cells.Results pShuttle-CMV-Hes1 shuttle plasmid and pAdeno-Hes1 plasmid were constructed successfully,with a general titer of 1.6 × 1011 PFU,Ad-Hes1 can be expressed in H9c2 myocardial cells,and its MOI value was 30.Conclusion Ad-Hes1 is successfully constructedand packaged,thus provide basis for further research on the protection effect of Hes1 on myocardium.
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Objective: To evaluate the association of glutathione S-Transferase P1 (GSTP 1) gene promoter methylation with the risk of prostate cancer. Methods: A computer-based online search was performed in PubMed, Embase, Cochrane Library, ISI Web of Science and China National Knowledge Infrastructure (CNKI) databases. According to the inclusion and exclusion criteria, the literatures about GSTP 1 gene promoter methylation and the risk of prostate cancer were selected. The difference in the incidence rate of GSTP 1 promoter methylation was compared between the prostate cancer tissues and the non-prostate cancer tissues, as well as in the blood or urine between the prostate cancer patients and the non-prostate cancer patients. The outcome measurements were odds ratio (OR ) with 95% confidence interval (95% CI ) for the association between the GSTP 1 gene promoter methylation and the risk of prostate cancer. The Meta-Analysis was performed using Stata 12.0 software. Results: A total of 19 studies including 1 889 samples were included in this Meta-Analysis. The result of the Meta-Analysis showed that GSTP 1 gene promoter methylation increased the risk of prostate cancer by 23.49 times [OR : 23.49 (95% CI : 13.14-42.01), P < 0.001]. Conclusion: GSTP 1 gene promoter methylation may increase the risk of prostate cancer.
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Objective: To study the expressions of enhancer of zeste homolog 2 (EZH2) and deleted in liver cancer-1 (DLC1) in breast cancer tissues and cell lines, and to explore their relationship. Methods: The expressions of EZH2 and DLC1 proteins in 120 cases of breast cancer tissues and 63 cases of para-cancerous tissues were detected by immunohistochemistry. The expression levels of EZH2 and DLC1 mRNAs and proteins in 20 cases of breast cancer tissues, 20 cases of the corresponding para-cancerous tissues, breast cancer MCF-7 and MDA-MB-231 cells and the normal mammary epithelial HBL100 cells were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The specific siRNA targeting EZH2 gene was transfected into MDA-MB-231 cells by liposome, then the expression levels of EZH2 and DLC1 mRNAs and proteins in MDA-MB-231 cells were detected again by realtime fluorescent quantitative PCR and Western blotting, respectively. Results: The positive expression rate of EZH2 in breast cancer tissues was significantly higher than that in corresponding para-cancerous tissues (P = 0.000). The positive expression rate of DLC1 in breast cancer tissues was significantly lower than that in corresponding para-cancerous tissues (P = 0.008). The expression rates of EZH2 and DLC1 were significantly correlated with tumor size and lymph node metastasis (both P < 0.05). The expression levels of EZH2 mRNA and protein in breast cancer tissues and breast cancer MCF7 and MDA-MB-231 cells were signifiicantly higher than those in corresponding para-cancerous tissues and normal breast epithelial HBL100 cells (all P < 0.01), but the expression levels of DLC1 mRNA and protein were lower than those in corresponding para-cancerous tissues and normal breast epithelial HBL100 cells (all P < 0.01). After EZH2 gene-silencing, the expression levels of DLC1 mRNA and protein in MDA-MB-231 cells were increased (both P < 0.05). Conclusion: There is a negative correlation between the expressions of EZH2 and DLC1 in breast cancer, and the inhibition of EZH2 expression can restore the expression of DLC1; which suggests that EZH2 maybe promote the occurrence and development of tumor by inhibiting the expression of DLC1.
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Objective:To investigate the effects of hematopoietic progenitor kinase 1 (HPK1) overexpression by construction of lentiviral vector on the proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells,and to elucidate its possible mechanism.Methods:The cells were infected with the lentivirus overxpressing HPK1,and the MCF-7-HPK1 and MDA-MB-231-HPK1 cell lines were stably expressed HPK1;each cell line was divided into three experimental groups:blank group (untreated),control group (empty vector) and HPK1-overexpression group.The expression levels of HPK1 mRNA and protein in breast cancer cells in each group were detected by RTPCR and Western blotting methods,respectively.The cell proliferation rate was detected by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry.Transwell assay was used to analyze the cell migration ability.Western blotting method was used to measure the expression levels of caspase 3,PTEN,MMP-9,MMP-2,Ki-67and HPK1 proteins.Results:Compared with blank groups and control groups,the expression levels of HPK1 mRNA and protein in the both cell lines in HPK1 overexpression groups were significantly up-regulated (P＜0.05),the proliferation rates were significantly decreased (P＜0.05) and the apoptotic rates were significantly increased (P＜0.05),the number of cells crossing matrigel was significantly reduced (P＜0.05),the cell cycle of MCF-7 was blocked in G1 phase (P＜0.05),the expression levels of caspase 3 and PTEN proteins in HPK1 overexpression group were significantly increased (P＜0.05),and the expression levels of MMP-2 and MMP-9 proteins were significantly decreased (P＜0.05).Conclusion:HPK1 overexpression can inhibit the proliferation and migration of MCF-7 and MDA-MB-231 cells and induce apoptosis,which may be related to the up-regulation of caspase 3 and PTEN and down-regulation of MMP-9,MMP-2 and Ki-67.
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Objective:To investigate the effects of hematopoietic progenitor kinase 1 (HPK1) overexpression by construction of lentiviral vector on the proliferation and apoptosis of breast cancer MCF-7 and MDA-MB-231 cells,and to elucidate its possible mechanism.Methods:The cells were infected with the lentivirus overxpressing HPK1,and the MCF-7-HPK1 and MDA-MB-231-HPK1 cell lines were stably expressed HPK1;each cell line was divided into three experimental groups:blank group (untreated),control group (empty vector) and HPK1-overexpression group.The expression levels of HPK1 mRNA and protein in breast cancer cells in each group were detected by RTPCR and Western blotting methods,respectively.The cell proliferation rate was detected by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry.Transwell assay was used to analyze the cell migration ability.Western blotting method was used to measure the expression levels of caspase 3,PTEN,MMP-9,MMP-2,Ki-67and HPK1 proteins.Results:Compared with blank groups and control groups,the expression levels of HPK1 mRNA and protein in the both cell lines in HPK1 overexpression groups were significantly up-regulated (P＜0.05),the proliferation rates were significantly decreased (P＜0.05) and the apoptotic rates were significantly increased (P＜0.05),the number of cells crossing matrigel was significantly reduced (P＜0.05),the cell cycle of MCF-7 was blocked in G1 phase (P＜0.05),the expression levels of caspase 3 and PTEN proteins in HPK1 overexpression group were significantly increased (P＜0.05),and the expression levels of MMP-2 and MMP-9 proteins were significantly decreased (P＜0.05).Conclusion:HPK1 overexpression can inhibit the proliferation and migration of MCF-7 and MDA-MB-231 cells and induce apoptosis,which may be related to the up-regulation of caspase 3 and PTEN and down-regulation of MMP-9,MMP-2 and Ki-67.
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Objective:To explore the expression of mammalian target of rapamycin (mTOR)and its relationgship with the prognosis of breast cancer,and to provide evidence-based basis for the using of mTOR inhibitor in the treatment of breast cancer.Methods: A systemical literature search was conducted based on the following databases:PubMed,EMBbase,Cochrane Library,ISI Web of Science,and CNKI up to November 24,2015.The outcome measures were hazard ratio (HR)with 95% confidence interval (CI) for the association between the mTOR expression and the prognosis of patients with breast cancer.The primary end points including disease-free survival (DFS ), and overall survival (OS ). STATA 12.0 was used to conduct the statistical analysis. Results:A total of seven cohort studies,1 758 patients were included. The risk of recurrence and metastasis of the breast cancer patients with positive expression of mTOR was 2.05 times of the patients with negative expression of mTOR (HR= 2.05, 95% CI: 1.01 - 4.13,P = 0.003);the risk of death in the breast cancer patients with positive expression of mTOR was 2.63 times of the patients with negative expression of mTOR (HR = 2.63, 95%CI:1.45-4.80,P = 0.736).Conclusion:The positive expression of mTOR can significantly increase the recurrence,metastasis and death risk of the patients with breast cancer.
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<p><b>OBJECTIVE</b>To construct rat N1ICD lentiviral over-expression vector (LV-N1ICD) and N1ICD lentivirus interference vector (LV-N1ICD-shRNA.</p><p><b>METHODS</b>With the rat cDNA as a template, the N1ICD fragment was amplified by PCR to construct pGC-FU-N1ICD-3Flag shuttle plasmid by directly clone. Four pairs of N1ICD-shRNA oligonucleotide sequences were syn- thesized to construct the GVC112-N1ICD-shRNA interference plasmid. pGC-FU-N1ICD-3Flag and GVC112 -N1ICD-shRNA plasmids were co-transfected into 293T cells to screen for the best interference plasmid in the 4 GVC112-N1ICD-shRNA plasmids by detecting Flag expression. pGC-FU-N1ICD-3Flag or GVC112-N1ICD-shRNA plasmid along with with pHelper 1.0 and pHelper 2.0 plasmids were co-transfect into 293T cells to package LV-N1ICD and LV-N1ICD-shRNA, and the virus titer was determined by real-time PCR and drug screening method, respectively. H9c2 cells infected with LV-N1ICD and LV- N1ICD-shRNA respectively were assessed for cell viability using CCK-8 assay.</p><p><b>RESULTS</b>pGC-FU-N1ICD-3Flag and GVC112- N1ICD-shRNA plasmid were verified by PCR, gene sequencing and Western blotting. Co-transfection of the plasmids with pHelper 1.0, and pHelper 2.0 plasmids into 293T cells obtained high-titer LV-N1ICD and LV-N1ICD-shRNA. LV-N1ICD was capable of promoting the cell viability and LV-N1ICD-shRNA produced an opposite effect.</p><p><b>CONCLUSION</b>The vectors LV-N1ICD and LV-N1ICD-shRNA have been successfully constructed and packaged, which have the biological functions of Notch1 signaling.</p>