LncRNA DERCNC in Hepatocellular Carcinoma with Cirrhosis Aggravates Tumor Proliferation by Targeting SOX9.

PURPOSE: This study aims to understand the role of cirrhosis in promoting hepatocellular carcinoma (HCC) progression by analyzing the differential expression of long noncoding RNAs (lncRNAs) between cirrhotic hepatocellular carcinoma (CHCC) and noncirrhotic hepatocellular carcinoma (NCHCC). METHODS: A transcriptional profile array was used to identify differentially expressed lncRNAs. Subsequently, a specific lncRNA was selected to evaluate the clinical significance, potential functions, regulatory targets, and pathways through both in vitro and in vivo experiments. RESULTS: The study identified a lncRNA, which we termed DERCNC, an acronym for Differentially Expressed RNA between Cirrhotic and Non-Cirrhotic HCC. DERCNC was significantly more highly expressed in CHCC than in NCHCC. Clinically, elevated levels of DERCNC expression were positively correlated with both the cirrhotic state and tumor stage and inversely correlated with tumor differentiation. Furthermore, high expression of DERCNC was associated with a poor prognosis for patients. Conditioned medium from the hepatic stellate cell (LX2) was found to enhance DERCNC expression, SOX9 expression, and tumor proliferation. Overexpression of DERCNC similarly promoted tumor proliferation and increased SOX9 levels. Conversely, DERCNC silencing resulted in the opposite effects. Moreover, the pro-proliferative function of DERCNC was reversible through the modulation of SOX9 expression. Further mechanistic studies revealed that DERCNC upregulated SOX9 by increasing the enrichment of H3K27ac modifications near the SOX9 promoter. CONCLUSION: In conclusion, DERCNC expression in CHCC has significant clinical implications and can aggravate tumor proliferation by targeting SOX9. This represents a novel mechanism by which cirrhosis promotes tumor progression.

High-intensity focused ultrasound combined with herpes simplex virus thymidine kinase gene-loaded ultrasound-targeted microbubbles improved the survival of rabbits with VX2 liver tumor.

BACKGROUND: To explore the anti-tumor effect of high-intensity focused ultrasound (HIFU) combined with herpes simplex virus thymidine kinase (HSV-TK) gene-loaded ultrasound-targeted microbubbles on VX2 rabbit liver tumors. METHODS: Seventy-five New Zealand white rabbits were randomly divided into five groups after the models of VX2 rabbit liver tumors were established: (a) HIFU group; (b) HIFU and HSV-TK group (HIFU + HSV-TK); (c) HIFU, HSV-TK and ultrasound group (HIFU + HSV-TK + US); (d) HIFU, HSV-TK gene-loaded microbubbles and ultrasound group (HIFU + HSV-TK-MBs + US); and (e) HSV-TK gene-loaded microbubbles and ultrasound group (HSV-TK-MBs + US). After 2 weeks of VX2 liver tumor implantation, rabbits in groups (a), (b), (c) and (d) received HIFU to establish rabbit models of residual tumor by ablating 80% of the tumor volume. After HIFU ablation, rabbits in different groups received MBs wrapped around HSV-TK or HSV-TK solution via marginal ear veins and/or local ultrasonic irradiation to the tumor. Six rabbits in each group were sacrificed 48 h after the corresponding treatment, and tumors were extracted for in vitro experiments. Thymidine kinase mRNA was detected by the real-time polymerase chain reaction. The green fluorescent protein expression in liver tumor was detected by western blotting and immunohistochemistry. Tumor cell apoptosis was detected by terminal deoxynucleotidyl transferase dUTP nick end labeling. The growth curves of VX2 liver tumors and survival curves of rabbits were compared. RESULTS: Forty-eight hours after treatment, TK mRNA and protein were the highest in the HIFU + HSV-TK + US + MBs group and the HSV-TK + US + MBs group (p < 0.05). At 48 h after treatment, the apoptotic index of tumor cells in HIFU + HSV-TK-MBs + US group was the highest (p < 0.05). Compared to other groups, HIFU combined with MBs wrapped HSV-TK suicide gene significantly inhibited tumor growth in vivo (p < 0.05) and prolonged the survival time of animals (p < 0.05). CONCLUSIONS: HIFU combined with HSV-TK gene-loaded ultrasound-targeted MBs significantly inhibited the growth of VX2 rabbit liver tumors in vivo and prolonged the survival time of the animals, providing a novel gene delivery method and a novel strategy for liver tumor treatment.

Field Investigation of Effect of Plants on Cracks of Compacted Clay Covers at a Contaminated Site.

Compacted clay covers (CCCs) are effective in restricting the upward migration of volatile organic compound (VOC) and semi-volatile organic compound (SVOC) vapors released mainly from unsaturated contaminated soils and hence mitigate the risks to human health. Desiccation cracking of CCCs would result in numerous preferential channels. VOC or SVOC vapors can prefereially migrate through the cracks and emit into the atmosphere, exposing threats to human health and surrounding environmental acceptors. This study presented results of comprehensive field investigation of desiccation crack distribution in CCCs, where four herbaceous plants were covered at the industrial contaminated site in. The plants included Trefoil, Bermuda grass, Conyza Canadensis, and Paspalum, and the corresponding planting areas were labeled as S1, S2, S3, and S4, respectively. The quantity and geometry parameters of the cracks including crack width, depth, and length, were investigated. The results showed that the cracks of the CCCs were mainly distributed in the areas of S3 (Conyza Canadensis) and S4 (Paspalum), where more cracks were formed when the degree of compaction (DOC) of the CCCs was less than 87%. In addition, the results revealed that: (1) no cracks were found in the area S1 (Trefoil); (2) the quantity, average width, average depth, average length, and maximal length of the cracks in the investigated areas followed S4 (Paspalum) > S3 (Conyza Canadensis) > S2 (Bermuda grass); (3) the maximal crack length in the area S2 (Bermuda grass) was the shortest, which was approximately one-seventh and one-eighth of those in the areas S3 (Conyza Canadensis) and S4 (Paspalum), respectively; and (4) the maximal width and depth of the cracks followed S3 (Conyza Canadensis) > S4 (Paspalum) > S2 (Bermuda grass).

Evaluation of the effectiveness of ex-situ stabilization for arsenic and antimony contaminated soil: Short-term and long-term leaching characteristics.

Ex-situ stabilization for As and Sb co-contaminated soil was conducted through an iron-based stabilizer, PFSC (a mixture of polymerized ferric sulfate (PFS) and hydrated lime (Ca(OH2)) with a dry mass ratio of 2:1). After field aging for one week, the stabilized contaminated soil was subjected to a horizontal vibration leaching test (HJ 557), Wenzel's sequential extraction, and a semi-dynamic leaching test (ANS 16.1). By assessing the cumulative fractions of As and Sb, the observed diffusion coefficients (Dobs) and leachability indices (LX) of metalloids released from the soil specimens were calculated. The PFSC ex-situ stabilization was effective to immobilize metalloids, and the As and Sb leached concentrations of stabilized contaminated soil samples were lower than remediation targets. Nonspecifically bound As and Sb in the stabilized contaminated soil samples decreased from 4.5 - 9.2 % to 1.5-2.5 % and from 2.2 - 5.8 % to 1.1-1.5 %, respectively. The mechanisms controlling the leaching behaviors of As and Sb included wash-off and diffusion and they were changed with the leaching interval. The mean Dobs of As and Sb released from stabilized contaminated soil specimen were 3.46 × 10-12 and 2.99 × 10-13 cm2 s-1, in the which were two orders of magnitude lower than that of untreated contaminated soil specimen. The mean LX of stabilized contaminated soil specimen for As and Sb releases were 11.40 and 12.83, respectively, indicating that the stabilized contaminated soil was acceptable for "controlled utilization".

Geoenvironmental properties of industrially contaminated site soil solidified/stabilized with a sustainable by-product-based binder.

This paper presents a study on utilizing a novel BCP binder, basic oxygen furnace slag (BOFS) activated with mixed calcium carbide residue (CCR) and phosphogypsum (PG), to solidify/stabilize heavy metals in industrial contaminated site soil. The effects of curing time and binder dosage on the geoenvironmental properties of the solidified/stabilized soil including soil pH, electrical conductivity, unconfined compressive strength, and leachability were tested and discussed. Chemical speciation of target heavy metals, pore-size distribution of treated soil, and phase identification of reaction products were analyzed to understand the mechanisms leading to the change of geoenvironmental properties. The results demonstrated that the addition of the BCP binder yielded remarkable increase in soil pH, unconfined compressive strength, and relative binding intensity index (IR) of target heavy metals including nickel (Ni) and zinc (Zn), while significantly decreased the electrical conductivity and leachability of contaminated soil. The IR value of heavy metals had a good linear relationship with the leached concentrations on a semi-logarithmic scale. The formation of heavy metal-bearing precipitates, absorptivity of calcium silicate hydrate (C-S-H), heavy metals encapsulation by C-S-H, and ion-exchange of heavy metals with ettringite (AFt) contributed to the immobilization of heavy metals in the solidified/stabilized soil.

[Effect of ultrasound microbubble carrying herpes simplex virus thymidine kinase on hepatocellular carcinoma in mice].

OBJECTIVE: To observe the effect of ultrasound microbubble carrying herpes simplex virus thymidine kinase hepatocellular carcinoma in mice. METHODS: Kunming mice were inoculated subcutaneously with H22 tumor cells. 40 male mice bearing subcutaneous hepatoma were randomized into 4 groups: PBS (group A), HSV1-TK (group B), HSV1-TK (group C), and microbubble carrying HSV1-TK (group D) were injected into the tail vein every 3 days. Mice in group C and D were exposed to ultrasound. The expression of TK protein was detected by western blot. Ganciclovir (GCV) was intraperitoneally injected at a dose of 100 mg x kg (-1) x d(-1) in group B, group C and group D. The tumor size was measured every 2 days. RESULTS: TK gene could be injected precisely into hepatocellular carcinoma with ultrasound monitor, and the expression of TK protein was found in all 4 groups. Expression in group D was higher than others (P < 0.05). The rate of tumor growth inhibition were 0 in group A, 3.90%+/-1.80% in group B, 22.70%+/-2.86% in group C, 41.25%+/-3.20% in group D (group B vs group C, P < 0.05; group D vs group C, P < 0.05; group D vs group B, P < 0.05). CONCLUSION: Ultrasound microbubble not only improve target gene therapy, but also enhance transfection efficiency.

Prognostic value of lymph nodes count on survival of patients with distal cholangiocarcinomas.

AIM: To evaluate the prognostic value of the number of retrieved lymph nodes (LNs) and other prognostic factors for patients with distal cholangiocarcinomas, and to determine the optimal retrieved LNs cut-off number. METHODS: The Surveillance, Epidemiology and End Results database was used to screen for patients with distal cholangiocarcinoma. Patients with different numbers of retrieved LNs were divided into three groups by the X-tile program. X-tile from Yale University is a useful tool for outcome-based cut-point optimization. The Kaplan-Meier method and Cox regression analysis were utilized for survival analysis. RESULTS: A total of 449 patients with distal cholangiocarcinoma met the inclusion criteria. The Kaplan-Meier survival analysis for all patients and for N1 patients revealed no significant differences among patients with different retrieved LN counts in terms of overall and cancer-specific survival. In patients with node-negative distal cholangiocarcinoma, patients with four to nine retrieved LNs had a significantly better overall (P = 0.026) and cancer-specific survival (P = 0.039) than others. In the subsequent multivariate analysis, the number of retrieved LNs was evaluated to be independently associated with survival. Additionally, patients with four to nine retrieved LNs had a significantly lower overall mortality risk [hazard ratio (HR) = 0.39; 95% confidence interval (CI): 0.20-0.74] and cancer cause-specific mortality risk (HR = 0.32; 95%CI: 0.15-0.66) than other patients. Additionally, stratified survival analyses showed persistently better overall and cancer-specific survival when retrieving four to nine LNs in patients with any T stage of tumor, a tumor between 20 and 50 mm in diameter, or a poorly differentiated or undifferentiated tumor, and in patients who were ≤ 70-years-old. CONCLUSION: The number of retrieved LNs was an important independent prognostic factor for patients with node-negative distal cholangiocarcinoma. Additionally, patients with four to nine retrieved LNs had better overall and cancer-specific survival rates than others, but the reason and mechanism were unclear. This conclusion should be validated in future studies.

Connexin32­mediated antitumor effects of suicide gene therapy against hepatocellular carcinoma: In vitro and in vivo anticancer activity.

Normal hepatocytes express connexin32 (Cx32), which forms gap junctions at cell­cell contact areas. The aim of the present study was to investigate whether Cx32 mediates the cell death­inducing effects of ultrasound microbubbles carrying the herpes simplex virus thymidine kinase (HSV­TK) suicide gene against hepatocellular carcinoma cells in vitro and in vivo. HepG2 cells were exposed to different concentrations of trans­retinoic acid (ATRA) in culture, to evaluate the intrinsic antitumor effect of ATRA. Detailed in­vitro and in­vivo investigations on the antitumor effects of ATRA via Cx32 mediation were performed, and the possible underlying mechanisms of action of the compound were then examined. The gene expression of HSV­TK transfected by ultrasound wave irradiation in the HepG2 cells was quantified using reverse transcription­quantitative polymerase chain reaction analysis. The effects on cell death were assessed using an MTT assay. The protein expression levels of Cx32 in ATRA­untreated or ATRA­treated tissues were quantified by immunohistochemical analysis and Western blot assays. The HSV­TK gene was successfully transfected into the HepG2 cell using ultrasound wave irradiation, and was stably expressed. Compared with the other groups, the HSV­TK gene group treated with ATRA exhibited an increased number of apoptotic cells (P<0.05) and improved tumor suppression (P<0.05). ATRA significantly increased the expression of Cx32 in the hepatoma tissues (P<0.01). The present study demonstrated that ATRA elevated the protein expression of Cx32 and enhanced the bystander effect of the HSV­TK/GCV suicide gene therapy system, which may provide a potential strategy for hepatocellular carcinoma treatment.

Diagnosis and cure experience of hepatolithiasis-associated intrahepatic cholangiocarcinoma in 66 patients.

BACKGROUND: The management of hepatolithiasis combined with intrahepatic cholangicarcinoma (IHHCC) remains a challenge due to poor prognosis. The aim of this study was to summarize our diagnosis and cure experience of IHHCC over the recent 10 years. METHODS: From January 1996 to January 2006, 66 patients with IHHCC were reviewed retrospectively. RESULTS: Of the 66 patients, 52 underwent surgical resection (radical resection in 38 and palliative in 14) and 8 patients abdominal exploration, while the other 6 cases received endoscopic retrograde biliary internal drainage and stent implantation. In this series, correct diagnosis of advanced stage was made during operation in 8 cases (8/60, 13.3%) and all of them (underwent unnecessary abdominal exploration, among them the positive rate of CA19-9 was 100%, and the positive rate of CEA was 87.6% (7/8), incidence rate of ascites was 100% and short-term significant weight loss was 100%, with median overall survival of only 4 months. CONCLUSION: Radical resection is mandatory for IHHCC patient to achieve long-term survival, the CT and MR imaging features of IHHCC being concentric enhancement. Patients with IHHCC have significant higher CA199 and significant higher CEA and short-term significant weight loss and ascites should be considered with advanced stage of IHHCC and unnecessary non-therapeutic laparotomies should be avoided.