ObjectiveTo compare clinical effect of percutaneous radiofrequency ablation (RFA) and open repeated hepatectomy (ORH) in treatment of liver cancer with late recurrence (recurrence time >12 months) and single tumor diameter ≤5 cm.MethodsThe patients with advanced intrahepatic recurrence after first operation for liver cancer in this hospital from January 2013 to December 2019 were retrospectively collected, who were treated with ORH (ORH group) or percutaneous RFA (RFA group) and met the inclusion criteria. The overall survival rate and disease-free survival rate of the two groups were compared after 1∶1 matching by propensity score matching (PSM), while the factors affecting survival were stratified.ResultsA total of 244 patients with recurrent liver cancer were collected, including 134 patients in the ORH group, 110 patients in the RFA group. The patients in the two groups were matched with 1∶1 by PSM, 90 patients in each group. The median overall survival time of the ORH group and the RFA group was 54 months and 45 months, respectively. There were no significant differences in the curves of cumulative overall survival and cumulative disease-free survival between the two groups (P=0.221, P=0.199). The incidence of severe complications in the ORH group was higher than that in the RFA group (10.00% versus 2.22%, P=0.029). A further subgroup analysis showed that the overall survival time of the ORH group was longer than that of the RFA group when the diameter of recurrent liver cancer was 3 to 5 cm (P=0.035), which had no significant differences for the patients with AFP (>400 μg/L or ≤400 μg/L), tumors number (single or multiple), and tumor diameter ≤3 cm between the two groups (P>0.05).ConclusionsPercutaneous RFA is effective and safe in treatment of advanced recurrent liver cancer, its overall survival and disease-free survival are similar to ORH treatment. However, when diameter of recurrent tumor is3–5 cm, ORH treatment has a advantage in prolonging survival time of patients.
Objective To explore the feasibility of recombinant adeno-associated virus (rAAV) as a vector for the gene therapy of liver cancer. Methods The rAAV/enhance green fluorescein protein (EGFP) recombinant was prepared by the routine method of two plasmids cotransfection.Results The experiment showed that one 10cm plate could produce 107-108 infection unit recombinant by the method of two plasmids cotransfection, and the transduction of HepG2 cell was increased with the increase of infection dosage of rAAV. About 100 multiplicity of infection (MOI) AAV vector could make all the tumor cell light. Conclusion Liver cancer cell can be efficiently transduced by rAAV, and AAV vector may be a valuable vector for the gene therapy of liver cancer.
Objective To investigate the value of contrast-enhanced ultrasonography in detection and diagnosis of small primary liver cancer. Methods SonoVue-enhanced ultrasonography were performed on 353 patients with 378 primary liver cancer, less than 3 cm in diameter. Enhancement patterns and enhancement phases of hepatic lesions on contrast-enhanced ultrasonography were analyzed and compared with the results of histopathology. Results In all hepatic tumors, 96.6% （365/378） lesions enhanced in the arterial phase. Among them, 317 (83.9%) tumors enhanced earlier than liver parenchyma and 48 (12.7%) tumors enhanced synchronously with liver parenchyma, and 342 (90.5%) tumors showed early wash-out in the portal and late phases. With regard to the enhancement pattern, 329 (87.0%) tumors presented whole-lesion enhancement, 35 (9.3%) to be mosaic enhancement and 14 (3.7%) to be rim-like enhancement. If taking the whole-lesion enhancement and mosaic enhancement in arterial phase as diagnotic standard for primary liver cancer on contrast-enhanced ultrasonography, the sensitivity was 92.9%(351/378), and if the earlier or synchronous enhancement of the tumor compared with liver parenchyma in arterial phase and the wash-out in portal phase were regarded as the stardand, the sensitivity was 87.3%(330/378). Conclusion Contrast-enhanced ultrasonography could display real-time enhancement patterns as well as the wash-out processes both in hepatic tumors and the liver parenchyma. It might be of clinical value in diagnosis of primary liver cancer based on the hemodynamics of hepatic tumors on contrast-enhanced ultrasonography.
To elucidate bcl-2 protein expression in hepatic carcinogenetic process and its relationship with apoptotic changes. bcl-2 protein was evaluated immunohistochemically while apoptosis was approached with terminal deoxynucleotidyl transferase (TdT)mediated dUTP nick end labeling (TUNEL) technique in 8 normal livers (NL), 17 liver cirrhosises (LC) and 77 hepatocellular carcinomas (HCC). The results showed that bcl-2 protein was expressed in 3 of 8 NLs(37.5%), 5 of 17 LCs(29.4%) and 7 of 77 HCCs(9.1%) with significant differences between group NL and HCC and between LC and HCC (Ｐ＜0.05). Apoptosis rates of 1.18±0.42%, 4.85±2.78%, 12.89±2.33% in NL, LC, HCC group respectively were demonstrated with significant differences among them (Ｐ＜0.01). Compared the bcl-2 expression with the apoptosis rate in this hepatocarcinogenetic process, reversed trends were presented. Conclusion: bcl-2 expression could be detected in NL, LC and HCC, and its decreasing expression was related to the inhibition attenuation of hepatocellular apoptosis in the process of hepatocarcinogenesis.
【Abstract】ObjectiveTo construct a recombinant eukaryotic expression vector for human endostatin in order to study the inhibitory effect of liposome-mediated endostatin gene on the growth of human liver carcinoma in nude mice. MethodsHuman endostatin cDNA including IL-2 secreting peptide was cloned into eukaryotic expression plasmid pcDNA3.0 to construct recombinant plasmid pCD-sEndo. pCD-sEndo plasmid was transferred into hepatocarcinoma cell line SMMC-7721 mediated by liposome Dosper, the expression and secretion of endostatin gene was detected by RTPCR and Western blot analysis. The suspension of SMMC-7721 cells was injected subcutaneously at the back of 32 nude mice to establish the model of human liver carcinoma. The mice were divided into 4 groups randomly, and injected with Dosper+pCD-sEndo, Dosper+pcDNA3.0, Dosper and physiological brine separately. Tumor volume was measured by stages. The mice were executed after the drug had been given for 1 week, then the microvessel density (MVD) of the tumor tissue was detected with immunohistochemical method and apoptotic index of tumor cells was measured by TUNEL-stain. ResultsThe eukaryotic expression vector pCD-sEndo was successfully constructed and was confirmed by enzyme digestion and sequence analysis. Expression of endostatin gene was detected in transfected SMMS-7721 cells by RTPCR in vitro, and endostatin protein was also detected in the supernatant of transfected SMMS-7721 cells by Western blot. In vivo study, the growth of human liver carcinoma was inhibited in the group injected with endostatin gene: the average volume of tumor in this group was significantly smaller than that in other groups （P＜0.05）; the average MVD in this group was 6.2±2.5, significantly less than that in the group injected with physiological brine (32.8±6.4), Dosper （27.8±6.4）, or Dosper+pcDNA3.0 (25.5±5.5), P＜0.05. The average apoptotic index of tumor cells in treatment group, brine group, Dosper group and Desper+pcDNA3.0 group was 24.5±7.3, 7.6±2.5, 9.5±3.0 and 11.2±3.6 respectively, it was evidently higher in the treatment group than in the latter three groups. ConclusionHuman endostatin mediated by cation liposome could decrease the microvessel number of implanted human hepatocarcinoma in nude mice. It could also accelerate apoptosis of tumor cells and inhibit growth of tumor.
The AC impedance of human hepatoma SMMC-7721 cells were measured in our laboratory by Agilent 4294A impedance analyzer in the frequency range of 0.01-100 MHz. And then the effect of hematocrit on electrical impedance characteristics of hepatoma cells was observed by electrical impedance spectroscopy, Bode diagram, Nyquist diagram and Nichols diagram. The results showed that firstly, there is a frequency dependence, i.e., the increment of real part and the imaginary part of complex electrical impedance (ΔZ', ΔZ″), the increment of the amplitude modulus of complex electrical impedance (Δ|Z*|) and phase angle (Δθ) were all changed with the increasing frequency. Secondly, it showed cell volume fraction (CVF) dependence, i.e., the increment of low-frequency limit (ΔZ'0, Δ|Z*|0), peak (ΔZ″p, Δθp), area and radius (Nyquist diagram, Nichols diagram) were all increased along with the electric field frequency. Thirdly, there was the presence of two characteristic frequencies: the first characteristic frequency (fC1) and the second characteristic frequency (fC2), which were originated respectively in the polarization effects of two interfaces that the cell membrane and extracellular fluid, cell membrane and cytoplasm. A conclusion can be drawn that the electrical impedance spectroscopy is able to be used to observe the electrical characteristics of human hepatoma cells, and therefore this method can be used to investigate the electrophysiological mechanisms of liver cancer cells, and provide research tools and observation parameters, and it also has important theoretical value and potential applications for screening anticancer drugs.