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Water inrush into coal mines from aquifers underlying coal seams often causes serious casualties and economic losses. The key to preventing the disaster is to discover a water inrush mechanism suitable for specific geological and hydrogeological conditions and apply reasonable control measures. A case of the Chensilou mine is studied in this paper. Complex geological and hydrogeological conditions, such as 12 aquifers in the floor and small distance between coal seam and aquifer, make the mining face in the synclinal basin have a great risk of water inrush. In addition, as an important way to prevent the disaster, grouting will aggravate the risk of water inrush from the floor. The slurry will drive groundwater in the limestone aquifers L8, even L7 and L6 along the horizontal (fracture zone in L11~L8) and vertical (Fs1 ~Fd1) water flow channel into the mining face and synclinal basin. A new water inrush mechanism driven by grouting is formed. In order to prevent this disaster, based on statistical law of hole deviation, the relative error of vertical depth and the angle between the borehole and the rock formation are obtained. Finally, an improved grouting method is proposed, which is useful to ensure the safe production of coal mine and reduce the cost of grouting.

The water inrush from karst collapse column (KCC) is a cascading, vicious cycle disaster caused by geological and mining activities, that can cause serious casualties and property losses. The key to preventing this risk is to study the mechanism of water inrush under confining pressure. Aiming at the investigationg the characteristics of the KCC named X1 in Chensilou mine, a series of methods, including connectivity experiments, water pressure monitoring tests in two side-walls, and numerical simulations based on plastic damage-seepage (PD-S) theory have been developed. The methods are used to test the security of the 2519 mining area, the damage thickness, pore water pressure, and seepage vector in the X1. The results indicate that the X1 has a certain water blocking capacity. In addition, with the decrease of confining pressure and increase of shear stress, deviatoric stress could cause the increase of permeability, the reduction of strength, and the reduction of pore water pressure in KCC. Therefore the increased effective stress in the rock will force the rock to become more fractured. Conversely, the broken rock could cause the change of stress, and further initiate new plastic strains, damage and pore water pressure until a new equilibrium is reached. This cascading water inrush mechanism will contribute to the exploitation of deep coal resources in complex geological and hydrogeological conditions.

Mining-induced groundwater inrush could result in serious economic losses and casualties. This disaster usually depends on geological structure, such as the karst collapse column (KCC). It could reduce the aquifuge thickness, causing a great risk of water bursting. In order to assess the impact of mining on the KCC, we have developed a complete set of methods, including three-dimensional seismic exploration, connectivity experiment, water injection experiment in two sidewalls, numerical simulation based on Drucker–Prager (D–P) elastoplastic damage theory. This method is used to test the size, type and the damage thickness of the KCC named DX1 in Sima mine, and the results indicate that the type of DX1 is good cementation. In addition, under the influence of the shear stress at the coal wall and the unloading at the goaf, a significant compressive damage zone with a large damage thickness will form at the sidewall of the KCC. As the roof collapses, the backfill is compacted and the maximum damage thickness gradually moves to the other side of the KCC. Eventually, an asymmetric inverted saddle-shaped damage distribution is formed with a maximum compression damage depth of 32.5 m. The results provide a basis for preventing water inrush from KCC.

The Hippo signaling pathway and the Notch signaling pathway are evolutionary conserved signaling cascades that have important roles in embryonic development of many organs. In murine liver, disruption of either pathway impairs intrahepatic bile duct development. Recent studies suggested that the Notch signaling receptor Notch2 is a direct transcriptional target of the Hippo signaling pathway effector YAP, and the Notch signaling is a major mediator of the Hippo signaling in maintaining biliary cell characteristics in adult mice. However, it remains to be determined whether the Hippo signaling pathway functions through the Notch signaling in intrahepatic bile duct development. We found that loss of the Hippo signaling pathway tumor suppressor Nf2 resulted in increased expression levels of the Notch signaling pathway receptor Notch2 in cholangiocytes but not in hepatocytes. When knocking down Notch2 on the background of Nf2 deficiency in mouse livers, the excessive bile duct development induced by Nf2 deficiency was suppressed by heterozygous and homozygous deletion of Notch2 in a dose-dependent manner. These results implicated that Notch signaling is one of the downstream effectors of the Hippo signaling pathway in regulating intrahepatic bile duct development.

The prediction of groundwater inflow in karst collapse pillar has an important impact on safety underground mining, and the occurrence of these groundwater disasters is likely to be controlled and decreased via establishing an accurate groundwater inflow prediction system. However, the relationship between groundwater inflow and the factors such as geological structure, hydrogeology, aquifer, groundwater pressure, groundwater-resisting layer, mining damage and so on can be highly nonlinear, so it is difficult to establish a suitable model using traditional mechanics methods to predict the groundwater inflow and inrush risk using time series data. It is appropriate to consider modelling methods developed in other fields in order to provide adequate models for rock behaviour on groundwater inflow, nonlinear grey Bernoulli model (NGBM) is a new grey prediction model which is a simple improvement of GM(1,1) together with Bernoulli differential equation. This paper presents a new parameter optimization scheme of NGBM using the genetic algorithm (GA). The power index r of Bernoulli differential equation and production coefficient of the background value are considered as decision variables, and the prediction mean absolute percentage error is taken as the optimization objective. Parameters optimization of NGBM was formulated as the combinatorial optimization problem and would be solved collectively using GA. The model can be optimized once the GA finds the optimal parameters of NGBM. NGBM with this parameter optimization algorithm is then applied in time series groundwater inflow prediction system. Results of long-term groundwater inflow prediction show that GA is an effective global optimization algorithm suitable for the parameter optimization of NGBM and the NGBM-GA model is suit for the groundwater inflow and inrush risk prediction. After grouting reconstruction, the groundwater inflow is decreased and there is no groundwater inrush risk in the process of mining.

A karst collapse pillar (KCP) is caused by the karst subsidence that occurs in carbonate rock distribution areas. Current KCP studies focus mainly on aspects such as the cause of formation, rule of development, groundwater inrush, and structural characteristics of the pillar and sidewall rock body. However, few existing studies focus on the influences of mining activity on the damage to KCP, its surrounding rocks, and the formation of the fracture zone, especially the mechanical behavior. In this paper, a numerical fast Lagrangian analysis of continua in three dimensions model was created to understand the mechanical state of a coal seam penetrated by a KCP during mining panel extraction. The numerical results suggest that (1) because of the structure defect, the stress field strongly depends on the mining activity, and the stress distribution varies in different parts of the KCP; (2) KCP and the surrounding rocks show dislocation when the working face passes the KCP, at which point shear failure may then occur, which will lead to a sidewall channel for groundwater inrush; and (3) as mining advances, the plastic zone in the KCP and its broken area develop gradually until the upper and lower plastic zones connect when the working face passes the KCP, which will cause a channel for groundwater inrush.

In order to study the permeability and water-resisting ability of the strata on the top of the Ordovician in Longgu Coal Mine, this paper tested the permeability and porosity of the strata, investigated the fracture and pore structure features of the strata, and identified the main channels which govern the permeability and water-resisting ability of the strata. The permeability of the upper, central and lower strata shows as 2.0504 × 10−3–2.782762 × 10−3, 4.1092 × 10−3–7.3387 × 10−3 and 2.0891 × 10−3–3.2705 × 10−3 μm2, respectively, and porosity of that is 0.6786–0.9197%, 0.3109–0.3951% and 0.9829–1.8655%, respectively. The results indicate that: (1) the main channels of the relative water-resisting layer are the pore throats with a diameter more than 6 μm; (2) the major proportion of pore throats in the vertical flow channel and the permeability first increases and then sharply decreases; (3) the fractures occurring from the top to 20 m in depth of the strata were filled and there occurred almost no fracture under the depth of 40 m; and (4) the ratio of turning point of the main flow channel in the strata on top of Ordovician can be used to confirm the thickness of filled water-resisting layers. Keywords: Relative water-resisting layer, Filling effect, Main channels of seepage, Pore structure

Although physiologic jaundice of neonates is common, persistent neonatal cholestasis is life-threatening and has multiple etiologies. Among these etiologies, biliary atresia (BA) requires rapid diagnosis and treatment. In diagnosing BA, the surgical pathologist must recognize subtle histologic changes, often with only a small core liver biopsy. To aid in the differential diagnosis of neonatal cholestasis, we investigated Yes-associated protein (YAP), a regulator of organ size and bile duct development. We examined whether a YAP immunostain can highlight emerging hepatobiliary epithelium in BA (n = 28) versus other causes of persistent cholestasis (non-BA; n = 15) and thus serve as a useful diagnostic marker in persistent neonatal jaundice. We show significantly (P < .01) more high-grade (

Groundwater inrush has an impartible relationship with geological structures such as karst collapse pillars (KCPs), which are widely distributed in North China. In order to study the mechanism of groundwater inrush from coal seam floor, the variable mass dynamics and nonlinear dynamics were introduced. A mechanical model–plug model is established to describe the behavior of water seepage flow in coal-seam-floor containing KCP. The study shows that: (1) If the mass of the KCP keeps steady, the water seepage velocity in the KCP and the surrounding rocks will reach a constant value soon; (2) if the mass of the KCP and the surrounding rocks increases by grouting, etc, the seepage velocity in the KCP and the surrounding rocks will reach its minimum value gradually, and (3) if the mass of the KCP and the surrounding rocks decreases by scouring, the flow velocity in the KCP and the surrounding rocks shows a monotone increase, the water flow may change into pipe flow, especially when a large number of the mass of the KCP and the surrounding rocks enters into goaf road, which may lead to instable flow and cause groundwater inrush.

The evaluation of the hydraulic properties evolution of granular sandstones in relation with groundwater inrush within faults is an important issue for mining engineering applications. This paper presents the results of an experimental investigation of small particle migration from granular sandstone samples under different original porosities, particle size compositions and water flow pressures. A new rock testing system has been setup to carry out the tests. Based on the results, it is observed that the overall permeability evolution during the tests can be divided into four different phases, including i) re-arrangement of large rock fragments, ii) water inrush with substantial particle migration, iii) continued moderate particles seepage, and iv) steady state water flow. The crushing of edges and corners of large rock fragments, and the evolution of the fracture network has mainly been observed in the first two phases of the tests. The results indicate that the migration of small particles has an essential effect on permeability and porosity increase during water inrush through fractured sandstone. The samples with higher original porosity, higher percentage of fine particles in their formation and under higher water flow pressures, achieve higher permeability and porosity values when the test is complete. Furthermore, using the measured data, the performances of a number of empirical models, for permeability evolution in fractured porous media, have been studied. The prediction results indicate that not all of the fractures in a sample domain contribute in small particle migration. There are parts of the fracture network that are not effective in particle flow, a sample with less original porosity, more fine particles and under lower water pressure shows less ineffective fractures. Therefore, using the concept of the effective porosity (fracture) is sufficient enough for the flow calculation.

Karst collapse pillar (KCP) is widespread in North China coalfields, where coal extraction above the Ordovician limestone aquifer is threatened by the abundant supply of water and a very high hydraulic pressure. KCP is composed of rock skeleton and fine fillings, which can be transferred under the effect of water flow, thus KCP usually functions as a channel for groundwater inrush. In order to investigate the mechanism of mining-induced groundwater inrush of KCP which was caused by fine fillings transfer, a self-designed particle transfer permeability testing system was used to test the crushed mudstones’ flow properties, which included fine filling particle transfer rate, porosity increase rate and permeability under the conditions of varying pore pressure, particle size mixture and compaction level (initial porosity). The tests indicate fine fillings transfer is the essential reason for mining-induced groundwater inrush (flow instability) of KCP. The flow properties changeability during fine particle transfer could be divided into four stages, i.e., initial flow stage, flow inrush stage, continued particle flow stage and stable flow stage, and flow inrush stage which is the key point to cause water inrush. Due to the crushing of edges and corners and the adjustment of the structure, the fluctuation of permeability–time relationship mainly distributed in the first two stages, which make a change to flow channel. Moreover, with the increasing of pore pressure, particle size mixture and initial porosity, the water inrush time and stable seepage time decreased more rapidly; the fine fillings, porosity and permeability increased gradually. The efficiency criteria analysis between measurement and calculation permeability shows an empirical equation can fit the relationship between porosity and permeability well, and not all of the pore structures were in flow; this means there was a part of pores that were invalid, but the effective porosity in flow can be treated as calculation value approximately.

Hepatocellular carcinoma and intrahepatic cholangiocarcinoma account for 95% of primary liver cancer. For each of these malignancies, the outcome is dismal; incidence is rapidly increasing, and mechanistic understanding is limited. We observed abnormal proliferation of both biliary epithelium and hepatocytes in mice after genetic manipulation of Yes-associated protein, a transcription coactivator. Here, we comprehensively documented Yes-associated protein expression in the human liver and primary liver cancers. We showed that nuclear Yes-associated protein expression is significantly increased in human intrahepatic cholangiocarcinoma and hepatocellular carcinoma. We found that increased Yes-associated protein levels in hepatocellular carcinoma are due to multiple mechanisms including gene amplification and transcriptional and posttranscriptional regulation. Survivin, a member of the inhibitors-of-apoptosis protein family, has been reported as an independent prognostic factor for poor survival in both hepatocellular carcinoma and intrahepatic cholangiocarcinoma. We found that nuclear Yes-associated protein expression correlates significantly with nuclear Survivin expression for both intrahepatic cholangiocarcinoma and hepatocellular carcinoma. Furthermore, using mice engineered to conditionally overexpress Yes-associated protein in the liver, we found that Survivin messenger RNA expression depends upon Yes-associated protein levels. Our findings suggested that Yes-associated protein contributes to primary liver tumorigenesis and likely mediates its oncogenic effects through modulating Survivin expression.

Water-inrush in mine is one of the mine disasters caused by mining. In order to assess the risk of roof water-inrush in Wuyang Coal Mine based on the geological material of the coal mine, we built numerical models for the roof fracture and seepage development rule by using RFPA2D and COMSOL respectively, to analyze the changes in fracture zone, stress, water pressure and seepage vector with the advancement of working face, and compared the results with the field investigated data. The numerical simulation results indicate that: (1) with the advancement of the working faces, the stress relief range and fracture zone in the overlying strata increased rapidly up to about 90 m, and then tended to remain constant, reaching a final height of about 95 m which agrees with the field investigation; (2) the seepage flow constantly increased with a larger flow volume both in the front and rear area, where the stress concentration are the most serious. Keywords: Fracture zone, Numerical simulation, Water inrush, Wuyang coal mine

The mining of shallow buried and thick coal seams may cause ground settlement, loss of surface water, and even soil desertification in arid&ndash, semi-arid climate zones. The key to protecting surface water resources through the technology of separated strata grouting is to study the evolution of the separation space under the key stratum (KS) with the change of strata. Aiming at the section of underground strata characteristics of F6202 and F6208 working faces, similar simulation tests under three typical geological conditions were carried out. Based on the experimental results, a theoretical calculation model was established, and the practice of separation grouting was carried out. The results show that, under the conditions of a thick coal seam with one KS, the structure formed by breaking strata below the KS was relatively unstable, and the separation space was relatively large, which is likely to cause serious ground settlement. More seriously, with the increase of the weakly consolidated layer and coal seam thickness, the increase of the ratio of rock to soil in the weakly consolidated layer, and the decrease of the thickness of the strata below the KS (two KS condition), the separation space was further increased. Based on the above analysis, combined with the real-time feedback data of the grout amount and pressure on hydraulic support, the position of the grouting borehole was determined, and the speed of mining advance was adjusted in time, thus effectively controlling the curvature of the ground settlement, protecting the integrity of the red clay layer near the surface, and consequently protecting water resources.

The checkpoint protein Rad9/Rad1/Hus1 heterotrimer (the 9-1-1 complex) is structurally similar to the proliferating cell nuclear antigen sliding clamp and has been proposed to sense DNA damage that leads to cell cycle arrest or apoptosis. Human (h) NEIL1 DNA glycosylase, an ortholog of bacterial Nei/Fpg, is involved in repairing oxidatively damaged DNA bases. In this study, we show that hNEIL1 interacts with hRad9, hRad1 and hHus1 as individual proteins and as a complex. Residues 290–350 of hNEIL1 are important for the 9-1-1 association. A significant fraction of the hNEIL1 nuclear foci co-localize with hRad9 foci in hydrogen peroxide treated cells. Human NEIL1 DNA glycosylase activity is significantly stimulated by hHus1, hRad1, hRad9 separately and the 9-1-1 complex. Thus, the 9-1-1 complex at the lesion sites serves as both a damage sensor to activate checkpoint control and a component of base excision repair.

As mining activity rapidly develops to deeper and deeper ground, there have been an increasing number of groundwater inrush events occurring in China. The Ordovician limestone karst collapse pillar (KCP), which contains a lot of crushed rocks, usually functions as a channel for groundwater inrush. The factors, such as the geological conditions, groundwater flow and extraction percentage of coal seam in underground mines, are the main causes of recompaction and permeability change of crushed rock particles in KCP. The high permeability of rock will cause large volumes of groundwater flow, thus posing a great threat of loss of lives in the mines. It is important to take into account the compaction and seepage behaviors of the crushed rocks in Ordovician limestone KCP. The MTS815.02 system and a self-designed water flow apparatus were used to investigate the effect of particle size distribution on compaction and seepage behavior of crushed limestone particle mixture. The Reynolds number calculation of particle mixture shows that the seepage has been influenced by non-Darcy flow. Testing results indicate that the effective porosity of crushed limestone sample is strongly influenced by compaction and particle size distribution. The effective porosity decreases with the increase in compaction and decrease in larger particle size, respectively. Particle crushing during compaction is a main cause of size 0–2.5 mm materials, whereas some fine particles are washed away due to the effect of water seepage, which is a main cause of weight loss. Non-Darcy seepage properties of the crushed limestone are strongly influenced by compaction and particle size distribution. In general, during the compaction, the permeability decreases while the non-Darcy coefficient increases with the decreasing of effective porosity. The effective porosity, particle crushing and seepage properties of crushed limestone are not only related to compaction levels and mixture sizes but also to style of arrangement and initial pore structure.

Background: For Hepatocellular carcinoma (HCC) and cholangiocarcinoma (CCA), the outcome is dismal; incidence is rapidly increasing, and mechanistic understanding is limited. YAP, the effector of Hippo signaling pathway, is an oncoprotein that promotes cell proliferation and inhibits apoptosis. Genetic manipulation of YAP results in biliary hamartoma and HCC. Therefore, we hypothesize that YAP is driver of liver cancer and a potential therapeutic target. Methods: Expression of YAP and its transcriptional targets GPC3 and Survivin was surveyed in normal human liver and primary liver cancer tissues. Efficacy of YAP inhibitor Vertepofin and Survivin inhibitor YM155 was tested in HCC and CCA cell lines with MTT assay. Results: Compared to the non-tumor tissue, nuclear YAP expression significantly increased in both HCC and ICC specimens. Nuclear YAP levels significantly correlate with nuclear Survivin levels in HCC and ICC tissues, but not with GPC3 in HCC tissues. We also found that verteporfin and YM155 supp...

The base excision repair DNA glycosylase MutY homolog (MYH) is responsible for removing adenines misincorporated into DNA opposite guanine or 7,8-dihydro-8-oxo-guanine (8-oxoG), thereby preventing G:C to T:A mutations. Biallelic germline mutations in the human MYH gene predispose individuals to multiple colorectal adenomas and carcinoma. We have recently demonstrated that hMYH interacts with the mismatch repair protein hMSH6, and that the hMSH2/hMSH6 (hMutSalpha) heterodimer stimulates hMYH activity. Here, we characterize the functional effect of two missense mutations (R227W and V232F) associated with hMYH polyposis that lie within, or adjacent to, the putative hMSH6 binding domain. Neither missense mutation affects the physical interaction between hMYH and hMSH6. However, hMYH(R227W) has a severe defect in A/8-oxoG binding and glycosylase activities, while hMYH(V232F) has reduced A/8-oxoG binding and glycosylase activities. The glycosylase activity of the V232F mutant can be partially stimulated by hMutSalpha but cannot be restored to the wild-type level. Both mutants also fail to complement mutY-deficiency in Escherichia coli. These data define the pathogenic mechanisms underlying two further hMYH polyposis-associated mutations.

Escherichia coli MutY is an adenine and weak guanine DNA glycosylase involved in reducing the mutagenic effects of 7,8-dihydro-8-oxoguanine (GO). MutY contains three structural domains: an iron-sulfur module, a six-helix barrel module with the helix-hairpin-helix motif, and a C-terminal domain. Here, we demonstrate that the mutant MutY(Δ26–134), which lacks the six-helix barrel domain, cannot complement the mutator phenotype of a mutY mutant in vivo. However, the mutant can still bind DNA and has weak catalytic activity at high enzyme concentrations. The mutant is a dimer in solution and assembled into two and multiple (up to five) complexes with 20- and 44-bp DNA fragments, respectively, in a concentration-dependent manner. Higher order complexes with DNA substrates containing A/GO mismatches were formed at lower protein concentrations than with the A/G mismatch and homoduplex DNA. Measurement of equilibrium binding using fluorescence anisotropy showed that the mutant protein retains some specificity for A/GO-containing DNA substrates and that the binding event is highly cooperative. This is consistent with the MutY structure determined, which indicates that GO specificity is contributed by both the six-helix barrel and C-terminal domains. The nonspecific binding of MutY(Δ26–134) to DNA suggests a model in which the specific binding of mismatched DNA by MutY involves sequential interactions, in which one MutY molecule scans the DNA and enhances binding of another MutY molecule to the A/GO mismatch.

Adenines mismatched with guanines or 7,8-dihydro-8-oxo-deoxyguanines that arise through DNA replication errors can be repaired by either base excision repair or mismatch repair. The human MutY homolog (hMYH), a DNA glycosylase, removes adenines from these mismatches. Human MutS homologs, hMSH2/hMSH6 (hMutSalpha), bind to the mismatches and initiate the repair on the daughter DNA strands. Human MYH is physically associated with hMSH2/hMSH6 via the hMSH6 subunit. The interaction of hMutSalpha and hMYH is not observed in several mismatch repair-defective cell lines. The hMutSalpha binding site is mapped to amino acid residues 232-254 of hMYH, a region conserved in the MutY family. Moreover, the binding and glycosylase activities of hMYH with an A/7,8-dihydro-8-oxo-deoxyguanine mismatch are enhanced by hMutSalpha. These results suggest that protein-protein interactions may be a means by which hMYH repair and mismatch repair cooperate in reducing replicative errors caused by oxidized bases.

Human chronic cholestatic liver diseases are characterized by cholangiocyte proliferation, hepatocyte injury, and fibrosis. Yes-associated protein (YAP), the effector of the Hippo tumor-suppressor pathway, has been shown to play a critical role in promoting cholangiocyte and hepatocyte proliferation and survival during embryonic liver development and hepatocellular carcinogenesis. Therefore, the aim of this study was to examine whether YAP participates in the regenerative response after cholestatic injury. First, we examined human liver tissue from patients with chronic cholestasis. We found more-active nuclear YAP in the bile ductular reactions of primary sclerosing cholangitis and primary biliary cirrhosis patient liver samples. Next, we used the murine bile duct ligation (BDL) model to induce cholestatic liver injury. We found significant changes in YAP activity after BDL in wild-type mice. The function of YAP in the hepatic response after BDL was further evaluated with liver-specific Yap conditional deletion in mice. Ablating Yap in the mouse liver not only compromised bile duct proliferation, but also enhanced hepatocyte necrosis and suppressed hepatocyte proliferation after BDL. Furthermore, primary hepatocytes and cholangiocytes isolated from Yap-deficient livers showed reduced proliferation in response to epidermal growth factor in vitro. Finally, we demonstrated that YAP likely mediates its biological effects through the modulation of Survivin expression. Conclusion: Our data suggest that YAP promotes cholangiocyte and hepatocyte proliferation and prevents parenchymal damage after cholestatic injury in mice and thus may mediate the response to cholestasis-induced human liver disease. (Hepatology 2012;56:1097–1107)

Purpose: To illustrate the prognostic significance of hedgehog (Hh) signaling in patients with hepatocellular carcinoma (HCC) and to evaluate the efficacy of a novel nanoparticle-encapsulated inhibitor of the Hh transcription factor, Gli1 (NanoHHI) using in vitro and in vivo models of human HCCs. Experimental Design: Patched1 (Ptch1) expression was detected in tumor tissue microarrays of 396 patients with HCC who underwent curative surgical resection during February 2000 to December 2002. Prognostic significance was assessed using Kaplan–Meier survival estimates and log-rank tests. The effects of NanoHHI alone and in combination with sorafenib were investigated on HCC cell lines. Primary HCC tumor growth and metastasis were examined in vivo using subcutaneous and orthotopic HCC xenografts in nude mice. Results: Elevated expression of Ptch1 in HCC tissues was significantly related to disease recurrence, as well as a shorter time to recurrence in patients with HCC. In vitro, NanoHHI significantly inhibited the proliferation and invasion of HCC cell lines. NanoHHI potently suppressed in vivo tumor growth of HCC xenografts in both subcutaneous and orthotopic milieus, and in contrast to sorafenib, resulted in significant attenuation of systemic metastases in the orthotopic setting. Furthermore, NanoHHI significantly decreased the population of CD133-expressing HCC cells, which have been implicated in tumor initiation and metastases. Conclusion: Downstream Hh signaling has prognostic significance in patients with HCC as it predicts early recurrence. Gli inhibition through NanoHHI has profound tumor growth inhibition and antimetastatic effects in HCC models, which may provide a new strategy in the treatment of patients with HCC and prevention post-operative recurrence. Clin Cancer Res; 18(5); 1291–302. ©2011 AACR.

Plant-derived polyphenols such as curcumin hold promise as a therapeutic agent in the treatment of chronic liver diseases. However, its development is plagued by poor aqueous solubility resulting in poor bioavailability. To circumvent the suboptimal bioavailability of free curcumin, we have developed a polymeric nanoparticle formulation of curcumin (NanoCurc™) that overcomes this major pitfall of the free compound. In this study, we show that NanoCurc™ results in sustained intrahepatic curcumin levels that can be found in both hepatocytes and non-parenchymal cells. NanoCurc™ markedly inhibits carbon tetrachloride-induced liver injury, production of pro-inflammatory cytokines and fibrosis. It also enhances antioxidant levels in the liver and inhibits pro-fibrogenic transcripts associated with activated myofibroblasts. Finally, we show that NanoCurc™ directly induces stellate cell apoptosis in vitro. Our results suggest that NanoCurc™ might be an effective therapy for patients with chronic liver disease.

The hippo pathway and its downstream mediator yes-associated protein 1 (YAP1) regulate mammalian organ size in part through modulating progenitor cell numbers. YAP1 has also been implicated as an oncogene in multiple human cancers. Currently, little is known about the expression of YAP1 either in normal human brain tissue or in central nervous system neoplasms. We used immunohistochemistry to evaluate nuclear YAP1 expression in the fetal and normal adult human brains and in 264 brain tumors. YAP1 was expressed in fetal and adult brain regions known to harbor neural progenitor cells, but there was little YAP1 immunoreactivity in the adult cerebral cortex. YAP1 protein was also readily detected in the nuclei of human brain tumors. In medulloblastoma, the expression varied between histologic subtypes and was most prominent in nodular/desmoplastic tumors. In gliomas, it was frequently expressed in infiltrating astrocytomas and oligodendrogliomas but rarely in pilocytic astrocytomas. Using a loss-of-function approach, we show that YAP1 promoted growth of glioblastoma cell lines in vitro. High levels of YAP1 messenger RNA expression were associated with aggressive molecular subsets of glioblastoma and with a nonsignificant trend toward reduced mean survival in human astrocytoma patients. These findings suggest that YAP1 may play an important role in normal human brain development and that it could represent a new target in human brain tumors.

OBJECTIVE To optimize the extracting process of ethanol extract with hypouricemic effect from Rhizoma Alismatis. METHOD Orthogonal design was utilized to optimize the preparing conditions including the concentration of ethanol, amount of ethanol, extraction times and extraction duration per time while alisol B 23-acetate was selected as the evaluation index. Both the alisol B 23-acetate and the serum uric acid levels were measured by HPLC. RESULT The ethanol extract had hypouricemic action. Only extraction times had significant effect on the content of alisol B 23-acetate in ethanol extract. CONCLUSION The optimal extraction process is to extract two times using sevenfold 70% ethanol and 1 h per time.

In eukaryotic cells, the cell cycle checkpoint proteins Rad9, Rad1, and Hus1 form the 9-1-1 complex which is structurally similar to the proliferating cell nuclear antigen (PCNA) sliding clamp. hMSH2/hMSH6 (hMutS alpha) and hMSH2/hMSH3 (hMutS beta) are the mismatch recognition factors of the mismatch repair pathway. hMutS alpha has been shown to physically and functionally interact with PCNA. Moreover, DNA methylating agent N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) treatment induces the G2/M cell cycle arrest that is dependent on the presence of hMutS alpha and hMutL alpha. In this study, we show that each subunit of the human 9-1-1 complex physically interacts with hMSH2, hMSH3, and hMSH6. The 9-1-1 complex from both humans and Schizosaccharomyces pombe can stimulate hMutS alpha binding with G/T-containing DNA. Rad9, Rad1, and Hus1 individual subunits can also stimulate the DNA binding activity of hMutS alpha. Human Rad9 and hMSH6 colocalize to nuclear foci of HeLa cells after exposure to MNNG. However, Rad9 does not form foci in MSH6 defective cells following MNNG treatment. In Rad9 knockdown untreated cells, the majority of the MSH6 is in cytoplasm. Following MNNG treatment, Rad9 knockdown cells has abnormal nuclear morphology and MSH6 is distributed around nuclear envelop. Our findings suggest that the 9-1-1 complex is a component of the mismatch repair involved in MNNG-induced damage response.

Escherichia coli MutY and MutS increase replication fidelity by removing adenines that were misincorporated opposite 7,8-dihydro-8-oxo-deoxyguanines (8-oxoG), G, or C. MutY DNA glycosylase removes adenines from these mismatches through a short-patch base excision repair pathway and thus prevents G:C-to-T:A and A:T-to-G:C mutations. MutS binds to the mismatches and initiates the long-patch mismatch repair on daughter DNA strands. We have previously reported that the human MutY homolog (hMYH) physically and functionally interacts with the human MutS homolog, hMutSα (Y. Gu et al., J. Biol. Chem. 277:11135-11142, 2002). Here, we show that a similar relationship between MutY and MutS exists in E. coli . The interaction of MutY and MutS involves the Fe-S domain of MutY and the ATPase domain of MutS. MutS, in eightfold molar excess over MutY, can enhance the binding activity of MutY with an A/8-oxoG mismatch by eightfold. The MutY expression level and activity in mutS mutant strains are sixfold and twofold greater, respectively, than those for the wild-type cells. The frequency of A:T-to-G:C mutations is reduced by two- to threefold in a mutS mutY mutant compared to a mutS mutant. Our results suggest that MutY base excision repair and mismatch repair defend against the mutagenic effect of 8-oxoG lesions in a cooperative manner.

The MutY homolog (MYH) can excise adenines misincorporated opposite to guanines or 7, 8-dihydro-8-oxo-guanines (8-oxoG) during DNA replication; thereby preventing G:C to T:A transversions. Germline mutations in the human MYH gene are associated with recessive inheritance of colorectal adenomatous polyposis (MAP). Here, we characterize one newly identified MAP-associated MYH missense mutation (R231L) that lies adjacent to the putative hMSH6 binding domain. The R231L mutant protein has severe defects in A/GO binding and in adenine glycosylase activities. The mutant fails to complement mutY-deficiency in Escherichia coli, but does not affect binding to hMSH6. These data support the role of the hMYH pathway in carcinogenesis.

The base excision repair carried out by bacterial MutY DNA glycosylase and eukaryotic MutY homolog (MYH) is responsible for removing adenines misincorporated into DNA opposite G and 7,8-dihydro-8-oxo-guanines (8-oxoG); thereby preventing G:C to T:A mutations. Escherichia coli MutY (EcMutY) can also remove adenines from A/C and A/5-hydroxyuracil and can remove guanines from G/8-oxoG mismatches at reduced rates. Thus, MutY has a minor role in reducing the mutagenic effects on G:C to A:T transitions and G:C to C:G transversions. The eukaryotic MYH can excise adenines misincorporated opposite GO, G, or C; remove 2-hydroxyadenines mispaired with A,G, and GO; excise G from G/GO mismatch weakly, thereby preventing G:C to T:A transversions. The in vitro and in vivo activities of MYH can be modulated by several proteins including apurinic/apyrimidinic endonuclease (APE1), proliferating cell nuclear antigen (PCNA), and mismatch recognition enzymes MSH2/MSH6. Recently, MYH has been shown to associate with the checkpoint proteins, Rad9, Rad1, and Hus1 (referred as the 9-1-1 complex). Thus, MYH-mediated base excision repair is coordinated with mismatch repair, DNA replication, cell-cycle progression, and DNA-damage checkpoints. Biallelic germ-line mutations in the human MYH gene are associated with recessive inheritance of multiple colorectal adenomas and carcinoma. MYH mutations can cause G:C to T:A mutations of the adenomatous polyposis coli (APC), K-ras, and other genes that control cellular proliferation in the colon.