Chinese Trauma Surgeon Association for management guidelines of vacuum sealing drainage application in abdominal surgeries-Update and systematic review / 中华创伤杂志(英文版)

Vacuum sealing drainage (VSD) is frequently used in abdominal surgeries. However, relevant guidelines are rare. Chinese Trauma Surgeon Association organized a committee composed of 28 experts across China in July 2017, aiming to provide an evidence-based recommendation for the application of VSD in abdominal surgeries. Eleven questions regarding the use of VSD in abdominal surgeries were addressed: (1) which type of materials should be respectively chosen for the intraperitoneal cavity, retroperitoneal cavity and superficial incisions? (2) Can VSD be preventively used for a high-risk abdominal incision with primary suture? (3) Can VSD be used in severely contaminated/infected abdominal surgical sites? (4) Can VSD be used for temporary abdominal cavity closure under some special conditions such as severe abdominal trauma, infection, liver transplantation and intra-abdominal volume increment in abdominal compartment syndrome? (5) Can VSD be used in abdominal organ inflammation, injury, or postoperative drainage? (6) Can VSD be used in the treatment of intestinal fistula and pancreatic fistula? (7) Can VSD be used in the treatment of intra-abdominal and extra-peritoneal abscess? (8) Can VSD be used in the treatment of abdominal wall wounds, wound cavity, and defects? (9) Does VSD increase the risk of bleeding? (10) Does VSD increase the risk of intestinal wall injury? (11) Does VSD increase the risk of peritoneal adhesion? Focusing on these questions, evidence-based recommendations were given accordingly. VSD was strongly recommended regarding the questions 2-4. Weak recommendations were made regarding questions 1 and 5-11. Proper use of VSD in abdominal surgeries can lower the risk of infection in abdominal incisions with primary suture, treat severely contaminated/infected surgical sites and facilitate temporary abdominal cavity closure.

Expression and clinical significance of nerve growth factor in primary liver cancer / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To determine the expression of nerve growth factor (NGF) in hepatic tissues and serum of patients with primary liver cancer (PLC), and to investigate the relationship of serum NGF levels with clinicopathological features of PLC.</p><p><b>METHODS</b>Hepatocellular carcinoma (HCC) samples and patient-matched tumor-adjacent liver samples were collected from 26 PLC patients to assess the mRNA and protein expressions of NGF by reverse transcription-PCR, western blotting (b-actin normalized), and immunohistochemistry. In addition, serum samples were collected from 40 PLC patients, 40 liver cirrhosis patients, 40 chronic hepatitis patients (including hepatitis B or C virus infections), and 30 healthy (normal) controls. The serum levels of NGF were measured by enzyme-linked immunosorbent assay. Intergroup differences were assessed by the t-test, and correlation with sex, age, presence of cirrhosis, tumor size, and TNM classification were assessed by the Kruskal-Wallis H test followed the Mann-Whitney U test.</p><p><b>RESULTS</b>HCC tissues showed higher mRNA and protein expressions of NGF than the corresponding tumor-adjacent non-HCC tissues. Hepatic NGF expression was mainly localized to the tumor cell cytoplasm. Serum NGF expression was significantly higher in PLC patients (33.86+/-16.11 pg/ml) and cirrhosis patients (20.57+/-9.73 pg/ml) than in normal controls (11.13+/-6.12 pg/ml) and chronic hepatitis patients (13.20+/-6.23 pg/ml) (P less than 0.01). Furthermore, when the PLC patients were stratified according to tumor size and TNM stage, the serum NGF level was found to be significantly higher in patients with tumors more than 5 cm (vs. less than 5 cm; U=83.000, P=0.002) or of TNM stage III/IV (vs. stage I/II; U=103.500, P=0.009).</p><p><b>CONCLUSION</b>Elevated expression of NGF in liver cancer tissues and serum of PLC patients is related with tumor size and TNM staging. These findings suggest that NGF may play a role in HCC tumorigenesis and/or that serum NGF may represent a prognostic marker of PLC.</p>

MiR-122 regulates the expression of PEG10 in hepatoma cell lines / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Our previous work indicated that overexpression of imprinting gene PEG10 is associated with malignant phenotype of hepatocellular carcinoma. The aim of this study is to explore whether disregulation of PEG10 leads to dysregulation of microRNAs.</p><p><b>METHODS</b>In silico analysis using TargetScan indicated that miR-122 could regulate the expression of PEG10. The expression of miR-122 in three hepatoma cell lines, Huh7, Hep3B and HepG2 and in primary human normal liver cell were compared using real time RT-PCR. After pre-miR-122 was transfected into HepG2 cell, the levels of PEG10 mRNA and protein were measured.</p><p><b>RESULTS</b>In silico analysis revealed that miR-122 could regulate the expression of PEG10. Real time RT-PCR indicated that miR-122 was not expressed in Hep3B and HepG2 cells, and only weakly expressed in Huh7 cells, but highly expressed in primary human normal liver cells. The expression of miR-122 was negatively correlated with the expression of PEG10. After pre-miR122 was transfected into HepG2, the mRNA level of PEG10 was not increased, whereas the protein level of PEG10 was increased.</p><p><b>CONCLUSION</b>miR-122 may be involved in regulation of PEG10 expression.</p>

A study on the apoptosis of hepatoma cells synergistically induced by plasmid-mediated anti-angiogenesis and immunopotentiation therapy / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effect and mechanism of apoptosis of implanted hepatoma cells in mice induced by eukaryotic plasmid-mediated anti-angiogenesis and immunopotentiation therapy.</p><p><b>METHODS</b>Mouse endostatin eukaryotic plasmid (pSecES) and mouse IL-12 (interleukin 12) eukaryotic plasmid (pmIL-12) were extracted and purified from E.coli. H22 hepatoma cells were inoculated into the leg muscles of 32 mice. The mice were divided into four groups with pSecES, pmIL-12, pSecES+pmIL-12 and pcDNA3.1 naked plasmid DNA injected into the tumor cell implantation sites of each group. Tumor formation and their weights were evaluated. Microvessel density, numbers of the infiltrating lymphocytes in the tumors and the apoptosis of tumor cells were assayed by microscopical examination of the CD31 and HE stained slides of the tumors and TUNEL assay.</p><p><b>RESULTS</b>The tumors of those with pSecES or pmIL-12 injections grew slower and with less microvessel density, more lymphocyte infiltration and with more apoptosis tumor cells compared with those with pcDNA3.1 injections. There was much more tumor cell apoptosis in the pSecES+pmIL-12 group (19.9+/-5.5 per 400x microscope field, P less than 0.05) than that in any other single plasmid injection group (400x microscopic field: pSecES 11.3+/-4.1, pmIL-12 14.6+/-3.2, pcDNA3.1 1.4+/-1.3).</p><p><b>CONCLUSIONS</b>Tumor cell apoptosis of the implanted hepatoma in mice can be induced by eukaryotic plasmid-mediated anti-angiogenesis and immunotherapy through inhibiting tumor angiogenesis and promoting tumor lymphocyte infiltration, by which the growth of the implanted hepatoma was inhibited.</p>

Inhibition of HSP70-2 expression by RNA interference induces apoptosis of human hepatocellular carcinoma cells / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To determine the effect of short hairpin RNA targeting HSP70-2 on the growth and apoptosis of hepatocellular carcinoma (HCC) cells, and to elucidate its possible mechanisms in mitochondria apoptotic pathway.</p><p><b>METHODS</b>Western blot and immunocytochemistry were used to determine the expression of HSP70-2 in HCC cells and normal hepatocytes. HepG2 and Huh-7 cells were cultured and transfected with HSP70-2 shRNA1 and shRNA2. Cell proliferation was examined by MTT. Cell apoptosis and mitochondria membrane potential were determined by flow cytometry. Western blot was used to analyze the expressions of apoptosis related proteins including Bax, Bcl-2, PARP, caspase-9 and caspase-3.</p><p><b>RESULTS</b>HSP70-2 was expressed at high levels in hepatocellular carcinoma (HCC) cells (SNU-449, HepG2, Huh-7, Hep3B) whereas there were very low levels in normal hepatocytes (L02). Using DNA vector-based RNA interference, we found that knockdown of HSP70-2 inhibited the growth of HCC cells through induction of mitochondria-dependent apoptosis. The mitochondria-dependent apoptosis induced by HSP70-2 knockdown was indicated by cytochrome c release from mitochondria, activation of caspase-9 and caspase-3, and loss of mitochondrial potential. Furthermore, knockdown of HSP70-2 resulted in up-regulation of the pro-apoptotic factor Bax and down-regulation of the pro-survival factor Bcl-2.</p><p><b>CONCLUSIONS</b>Our results indicated that HSP70-2 down-regulation induces apoptosis of HCC cells through the mitochondrial apoptotic pathway, highlighting the importance of HSP70-2 in survival of HCC cells and maintenance of liver function.</p>

Relationship between death receptor 5 and apoptosis in hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the antitumor efficacy of death receptor 5, its ligand (TRAIL) and DR5mAb in human hepatocellular carcinoma.</p><p><b>METHODS</b>Expression of DR5 in the HCC cell lines HepG2, SMMC 7721 and normal human liver cell line LO2 was measured at mRNA and protein level by semi-quantitative RT-PCR and Western blot, respectively. MTT method was used to measure the cell viability and flow cytometry assay was used to detect apoptosis so as to observe the inhibitory effect of TRAIL and DR5mAb on HCC cells.</p><p><b>RESULTS</b>Death receptor 5 was highly expressed in the HCC cell lines, but rarely expressed in normal human liver cell line (P < 0.01). With the increase of TRAIL concentration, the cell viability of HCC cells decreased gradually. However, when the concentration of TRAIL was above 1000 ng/ml, HCC cells were resistant to TRAIL, but still sensitive to DR5mAb. After incubation with DR5mAb (1000 ng/ml) for 24 h, the rate of apoptosis in HCC cells reached to 52.45% +/- 0.57%, which was higher than that incubated with TRAIL under the same condition (14.74% +/- 0.48%) (P < 0.05). The cell viability of normal human liver cell line treated with TRAIL tended to decline with the increase of the concentration, which was significantly different from that of matched control group. But DR5mAb had little effect on normal human liver cell line.</p><p><b>CONCLUSION</b>Death receptor 5 as a target plays an important role in the course of HCC apoptosis induction. Agonistic monoclonal antibody specific for human DR5 can selectively and effectively kill hepatocellular carcinoma cells in vitro, while is not harmful to normal human hepatocytes. It reveals that DR5mAb might provide a new direction in hepatocellular carcinoma treatment research.</p>

Establishment of a new HBV genotyping method with PCR-RBD and its application / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Using PCR-RDB to establish a new method for HBV genotyping, and to survey the distribution of HBV genotypes in the Foshan area.</p><p><b>METHODS</b>Biotin-labeled primers for amplification of HBV region X (nt1550-1789) were used to amplify extracted HBV DNA. HBV was genotyped by hybridization of the PCR products with immobilized specific probes (genotype A to F) on C membrane. Color development was achieved by adding POD and TMB. A judgment was made according to color reactions. The reliability of this new method was verified by gene sequencing. 300 samples of HBV DNA-positive sera from the Foshan area were genotyped using this assay.</p><p><b>RESULTS</b>Of the 300 sera genotyped by PCR-RBD, 147 (49.0%) cases were genotype B, 136 (45.3%) were genotype C, 1 (0.3%) genotype D, and 12 (4.0%) were mixtures of genotype B and C, and 4 (1.3%) were mixtures of genotype C and D. No genotype A, E or F were found. The results of PCR-RDB genotyping were consistent with the results obtained with sequence analysis.</p><p><b>CONCLUSION</b>This newly established HBV genotyping system proved to be sensitive, specific, precise and economic, and should be suitable for clinical practice and epidemic study. The results of HBV genotyping show that genotype B and C are the predominant genotypes in the Foshan area.</p>

Hammerhead ribozyme against human telomerase catalytic subunit (hTERT) induced apoptosis of liver cancer cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To construct vector pEGFP-C1-hTERT-ribozyme (pGTRz-U6) and its mutant (pGTmRz-U6) against hTERT containing U6 promoter, then transfect them into human liver cancer cell line SMMC7721 to observe the action of the human telomerase catalytic subunit (hTERT) hammerhead ribozyme on proliferation and apoptosis of human liver cancer cell SMMC7721.</p><p><b>METHODS</b>Eukaryotic expressing vector pGTRz-U6 and mutant pGTmRz-U6 were constructed and transfected into SMMC7721 using Lipofectamine2000 Reagent, with pEGFP-C1 as the control group. After strict screening by G418, positive clones were cultured; the amount of expression of ribozyme and hTERT was detected by RT-PCR; cell proliferation by MTT; telomerase activity by TRAP and silver staining assay; cell apoptosis by FCM.</p><p><b>RESULTS</b>We found that the two ribozymes were expressed persistently in SMMC7721; different expression levels (P < 0.01) of hTERT among SMMC7721-Rz, SMMC7721-mRz and SMMC7721-pEGFP-C1 was exhibited by the analysis of variance with SPSS software. The difference between SMMC7721-Rz and the others is significant in t-test (P < 0.01), while there was no difference between SMMC7721-mRz and SMMC7721-pEGFP-C1 (P > 0.05). With the advance of cell division, telomerase activities of the cells treated by SMMC7721-Rz and SMMC7721-mRz decreased gradually, and the percentage of apoptosis of the cells transfected with Rz and mRz increased gradually. The apoptosis percentage of 7PDS SMMC7721-Rz was 29.86%, while those of SMMC7721-mRz and SMMC7721-pEGFP-C1 were 9.87% and 3.36%, respectively.</p><p><b>CONCLUSION</b>The apoptosis level of SMMC7721 induced by hTERT ribozyme increases as cells divide, and this ribozyme maybe a potential approach for liver cancer gene therapy.</p>

Therapeutic effect of secretive endostatin eukaryotic expressing plasmid on mouse hepatoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To construct and express secretive endostatin eukaryotic plasmid for treatment of hepatoma.</p><p><b>METHODS</b>Mouse Igk signal peptide sequence was synthesized and cloned into pcDNA3.1 with endostatin gene. The supernant of BHK-21 transfected with recombinant was used to culture ECV304. The proliferation of latter was evaluated by MTT assay. H22 was inoculated intramusclely, then naked DNA of endostatin plasmid was injected into the inoculation site. Tumors were dissected and weighted after treatments. All data was analyzed by SPSS10.0.</p><p><b>RESULTS</b>The supernant of BHK-21 transfected with recombinant can inhibit the proliferation of ECV304 by 29.2%. Tumor weight lighter after injected with naked pSecES (1.34 g+/-0.96g) compared with naked pcDNA3.1 (2.70g+/-0.82g) and saline (3.73g+/-1.41g).</p><p><b>CONCLUSION</b>The endostatin eukaryotic plasmid was constructed and it can be used for gene therapy on hepatoma.</p>