Objective To establish hepatocellular carcinoma (HCC) cell lines which olig-expressed IGF1R gene stably． Methods An eukaryotic expressing vector pSUPER-IGF1R-siRNA that could block IGF1R expressing was transferred into hepatocellular carcinoma cell lines SMMC7721 and Hep3B with Lipofectamine 2000 reagents． After transferred, cells were selected with G418 to obtain positive clones. The expressions of IGF1R, cyclin D1 and cyclin B1 were detected by RT-PCR and Western-blot． Cell growth curve were painted. Results Two cell lines clones were screened olig-expressing IGF1R gene stably． The experimental cell lines grew more slowly than control cell lines and the expression of cyclin D1 decreased (P＜0.05). Conclusion The HCC cell lines for olig-expressing IGF1R gene stably are established successfully．The plasmid pSUPER-IGF1R-siRNA can inhibit the growth of SMMC7721 and Hep3B cell lines, and the expression of cyclin D1.
【Abstract】Objective To investigate the expression of tumor growth tactor β1 (TGFβ1) and p27 in gallbladder carcinoma and their relation to the development of the carcinoma. Methods The expression of TGF-β1 and p27 in 36 cases of gallbladder carcinoma was detected by SP immunohistochemical staining. Twenty cases of chronic cholecystitis were collected as control. Results The positive rate of TGF-β1 （63.9%） was higher than that of the control （10.0%），P＜0.05， and the positive rate of p27 (47.2%) was lower than that of the control 100%(P＜0.05). The positive rate of TGF-β1 was significantly higher in metastasis or Nevin Ⅳ～Ⅴ stage cases than that of non-metastasis or Nevin Ⅰ～Ⅲ stage cases 33.3% (P＜0.05). The positive rate of p27 was statistically higher in moderate and highly differentiation (60.9%), nonmetastasis (75.0%) or Nevin’s Ⅰ～Ⅲ stage (75.0%) cases than those of poor differentiation (23.0%), metastasis (33.3%) and Nevin Ⅳ～Ⅴ stage (33.3%) cases (P＜0.05). The expression of p27 was negatively correlated with that of TGF-β1(r=-0.4473，P＜0.05). There was significant difference in survival time between patients with TGF-β1 positive and TGF-β1 negative（P＜0.05）. The difference was also found between patients with p27 positive and p27 negative. Conclusion The upregulation of TGF-β1 and downregulation of p27 in gallbladder carcinoma indicates the imbalance of TGF-β1/p27 system, which may play a role in the carcinogenesis and predict the malignant behaviors of the carcinoma.
目的 探讨子宫内膜癌中雌激素受体（ER）、孕激素受体（PR）和p53的表达及其临床意义。 方法 1994年1月-2009年11月，应用免疫组织化学Envision法检测52例子宫内膜癌中ER、PR和p53的表达情况并进行统计学分析。 结果 ER、PR和p53阳性表达率分别为51.9%、50.0%和46.2%。ER、PR的阳性表达率与癌组织的细胞分化程度有关（Plt;0.05）。随着组织学分级的增高，ER、PR阳性表达率逐渐降低（在Ⅰ、Ⅱ和Ⅲ级子宫内膜癌中，ER阳性表达率分别为64.0%、56.3%和18.2%， PR阳性表达率分别为72.0%、37.5%和18.2%）。随着组织学分级的增高及淋巴结转移，p53的阳性表达率逐渐增高（Plt;0.05），p53表达与ER、PR表达有关（Plt;0.05）。 结论 ER、PR和p53的表达与子宫内膜癌组织学分级和生物学行为密切相关，其测定对评估子宫内膜癌预后，指导临床治疗具有重要意义。
Objective To study the interferencing and anti-tumor effects of lentiviral vector of siRNA targeting IGF1R and EGFR gene of the liver cancer cell. Methods The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed, synthesized and connected to the pLVTHM vector, named pLVTHM-IGF1R, into whom the EGFR-siRNA expression frame containing H1 promotor synthesized by RT-PCR was cloned to generate pLVTHM-IGF1R-EGFR-siRNA. The 293T cells were cotransfected by 3 plasmids of pLVTHM-IGF1R-EGFR-siRNA, psPAX2 and pMD2G to enclose LVTHM-IGF1R-EGFR-siRNA, which was amplified in large amount and purified by caesium chloride density gradient centrifugation for measurement of virus titer. SMMC7721 cells infected by LVTHM-IGF1R-EGFR-siRNA were infection group, the untreated SMMC7721 cells and blank vector plasmid LVTHM were two control groups (SMMC7721 cell group and blank vector group). The effect of LVTHM-IGF1R-EGFR-siRNA on IGF1R and EGFR expressions of SMMC7721 cells were detected by RT-PCR and Western blot. The antitumor potential of LVTHM-IGF1R-EGFR-siRNA to SMMC7721 cells was evaluated by Cell Counting Kit-8 assay for cell growth and TUNEL for apoptosis respectively. Results LVTHM-IGF1R-EGFR-siRNA was constructed successfully. Functional pfu titers of LVTHM-IGF1R-EGFR-siRNA was 4.58×109 pfu/ml. Protein and mRNA expression of IGF1R and EGFR of infection group were less than those of blank vector group and SMMC7721 cell group （P＜0.05）, LVTHM-IGF1R-EGFR-siRNA was more effective to inhibit the proliferation and promote apoptosis of SMMC7721 cells （P＜0.05）. Conclusion LVTHM-IGF1R-EGFR-siRNA expressing IGF1R-EGFR-siRNA can inhibit the expression of IGF1R and EGFR, and may be used for further investigation of gene therapy of liver cancer.
ObjectiveTo compare the clinical effect of laparoscopic Heller myotomy (LHM) combined with Dor fundoplication and peroral endoscopic myotomy (POEM) in treatment of patients with achalasia.MethodsThe clinical data of 67 patients with achalasia from January 2014 to December 2018 in the Affiliated Hospital of Xuzhou Medical University were retrospectively analyzed. Among them, 19 patients received the LHM combined with Dor fundoplication (LHM group), 48 patients received the POEM (POEM group). The clinical efficacy and safety of the two groups were compared.ResultsThere were no significant differences in the baseline data such as the gender, age, course of disease, body mass index, preoperative Eckardt score, preoperative maximum diameter of esophagus, and previous treatment history between the two groups (P>0.05). There were no significant differences in the operation time, bleeding volume, the Eckardt points at 3 and 12 months after operation, the decrease degree of maximum diameter of esophagus, complications (except for gastroesophageal reflux, P=0.029), and recurrence rate between the two groups (P>0.05). The total hospitalization time, postoperative hospitalization time, and total hospitalization costs of the POEM group were lower than those of the LHM group (P<0.05).ConclusionsBoth LHM and POEM could effectively relieve clinical symptoms, short-term efficacy and safety of the two kinds of operations are similar. Postoperative recovery of POEM is fast and hospitalization cost is less, but incidence of gastroesophageal reflux is higher.
Objective To investigate the impact of laparoscopic versus. open hepatic resection for liver cancer on clinical rehabilitation and humoral immune function in patients organism. Methods Forty-four patients of laparoscopic and open left-lateral sectionectomy from January 2010 to June 2012 were selected， including 22 patients of laparoscopy group and 22 patients of conventional laparotomy group. The levels of IgG， IgA， IgM， C3， C4， C reactive protein （CRP）， IL-2， IL-6， and TNF-α in peripheral blood of patients on the last day before operation， first day and 5th day after operation were determinated by using ELISA assay. At the same time， the operative time， intraoperative bleeding， hospitalization time， and complications after operation between two groups were compared. Results The postoperative analgesic using time， first time eating， and hospitalization time in laparoscopic group were （1.9±0.8） days， （2.2±0.5） days， and （6.3±1.3） days， respectively， they were shorter than that in conventional laparotomy group （P＜0.05）. The operative time， intraoperative bleeding， complication rate， and mortality in two groups were not significant differences（P＞0.05） . Compared with before operation， the levels of C3， C4， IgA， IgG， IgM， and IL-2 on the first day after oper-ation in two groups were obviusly reduced， the levels of CRP， IL-6， and TNF-α on the first day after operation in two groups were significantly increased. The levels of C3， C4， lgA， IgG， lgM， and IL-2 on the first day after operation in conventional laparotomy group were significantly decreased than that in laparoscopic group （P＜0.05）. On the 5th day after operation， the levels of C3， C4， lgA， IgG， lgM， and IL-2 of laparoscopy group increased， the levels of CRP， IL-6，and TNF-α were reduced，that were no difference compared with before operation. Compared with before operation，the levels of C3， C4， lgA， IgG， lgM， and IL-2 of conventional laparotomy group were still at a low level state， and the levels of CRP， IL-6， and TNF-α were still at a high level state on the 5th day after operation. Conclusions Laparoscopic resection of liver cancer after operation， the patients’ recovery are quickly， and the impact on humoral immune function of laparoscopic radical resection for liver cancer patients is significantly less than that conventional laparotomy.
Objective To investigate the risk factors associated with the formation of parastomal hernia after Miles operation, and to provide scientific evidence for the prevention of parastomal hernia. Methods The clinical data of 218 patients with rectal cancer who undergoing Miles operation in the Affiliated Hospital of Xuzhou Medical University between May 2011 and May 2015 were analyzed retrospectively. Fifteen possible factors affecting the occurrence of parastomal hernia were selected. The fifteen related factors were analyzed by single factor analysis and two elements logistic regression analysis. Results A total of 55 patients with parostomy hernia occurred in 218 patients during follow-up period, and the incidence of parastomal hernia was 25.23%. Univariate analysis showed that age, thickness of subcutaneous fat in abdomen, BMI, stoma pathway, and hypertension were related to the formation of parastomal hernia after Miles operation (P<0.05). Further two elements logistic regression analysis showed that, advanced age, thickness of subcutaneous fat in abdomen, BMI>25 kg/m2, and transperitoneal were independent risk factors for the formation of parastomal hernia after Miles operation (P<0.05). Conclusion For patients with risk factors, it is very important to choose optimum stoma pathway during Miles operation to avoid the occurrence of formation of parastomal hernia after Miles operation.