Regulation mechanism of endoplasmic reticulum stress on metabolic enzymes in liver diseases.

The endoplasmic reticulum (ER) plays a pivotal role in protein folding and secretion, Ca2+ storage, and lipid synthesis in eukaryotic cells. When the burden of protein synthesis and folding required to be handled exceeds the processing capacity of the ER, the accumulation of misfolded/unfolded proteins triggers ER stress. In response to short-term ER stress, the unfolded protein response (UPR) is activated to allow cells to survive. When ER stress is severe and sustained, it typically provokes cell death through multiple approaches. It is well documented that ER stress and metabolic deregulation are functionally intertwined, both are considered contributing factors to the pathogenesis of liver diseases, including non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease (ALD), ischemia/reperfusion (I/R) injury, viral hepatitis, liver fibrosis, and hepatocellular carcinoma (HCC). Hepatocytes are rich in smooth and rough ER, which harbor metabolic enzymes that are capable of sensing alterations in various nutritional status and external stimuli. Extensive research has focused on the molecular mechanism linking ER stress with metabolic enzymes. The purpose of this review is to summarize the current knowledge regarding the effects of ER stress on metabolic enzymes in various liver diseases and to provide potential therapeutic strategies for chronic liver diseases via targeting UPR.

Metabolism and pharmacokinetic study of deuterated osimertinib.

Osimertinib is a highly selective third-generation irreversible inhibitor of epidermal growth factor receptor mutant, which can be utilized to treat non-small cell lung cancer. As the substrate of cytochrome P450 enzyme, it is mainly metabolized by the CYP3A enzyme in humans. Among the metabolites produced by osimertinib, AZ5104, and AZ7550, which are demethylated that is most vital. Nowadays, deuteration is a new design approach for several drugs. This popular strategy is deemed to improve the pharmacokinetic characteristics of the original drugs. Therefore, in this study the metabolism profiles of osimertinib and its deuterated compound (osimertinib-d3) in liver microsomes and human recombinant cytochrome P450 isoenzymes and the pharmacokinetics in rats and humans were compared. After deuteration, its kinetic isotope effect greatly inhibited the metabolic pathway that produces AZ5104. The plasma concentration of the key metabolite AZ5104 of osimertinib-d3 in rats and humans decreased significantly compared with that of the osimertinib. This phenomenon was consistent with the results of the metabolism studies in vitro. In addition, the in vivo results indicated that osimertinib-d3 had higher systemic exposure (AUC) and peak concentration (Cmax ) compared with the osimertinib in rats and human body.

MicroRNA-199b-5p attenuates TGF-ß1-induced epithelial-mesenchymal transition in hepatocellular carcinoma.

BACKGROUND: Accumulating evidence indicates that N-cadherin is a cell adhesion molecule that has critical roles in tumour progression. However, the role of N-cadherin in hepatocellular carcinoma (HCC) remains controversial. METHODS: This study aims to investigate the expression status of N-cadherin and its molecular mechanisms in HCC. RESULTS: The expression of N-cadherin was markedly overexpressed in HCC tissues and cell lines. We identified that miR-199b-5p binds to the 3'-UTR of N-cadherin mRNA, thus decreasing N-cadherin expression in HCC cells. We also found the downregulation of miR-199b-5p in HCC specimens, which was inversely correlated with N-cadherin upregulation, predicted poor clinical outcomes in HCC patients. Next, we determined that miR-199b-5p overexpression promoted cell aggregation, suppressed cell migration and invasion in HCC cells, and inhibited xenografts tumour metastasis in nude mice. Moreover, we demonstrated that miR-199b-5p attenuated TGF-ß1 induced epithelial-mesenchymal transition (EMT) -associated traits, while its effects could be partially reversed by N-cadherin restoration. Finally, we examined that N-cadherin downregulation or miR-199b-5p overexpression suppressed TGF-ß1-induced Akt phosphorylation, and inhibition of PI3K/Akt pathway blocked TGF-ß1-induced N-cadherin overexpression in HCC cells. CONCLUSIONS: Our data demonstrate that N-Cadherin was markedly overexpressed and miR-199b-5p was significantly downregulated in HCC. MiR-199b-5p exerts inhibitory effects on EMT, and directly targets N-cadherin in HCC, supporting the potential utility of miR-199b-5p as a promising strategy to treat HCC. Also, a positive regulatory loop exists between N-cadherin and Akt signalling represents a novel mechanism of TGF-ß1-mediated EMT in HCC cells.

Nanovalved Adsorbents for CH4 Storage.

A novel concept of utilizing nanoporous coatings as effective nanovalves on microporous adsorbents was developed for high capacity natural gas storage at low storage pressure. The work reported here for the first time presents the concept of nanovalved adsorbents capable of sealing high pressure CH4 inside the adsorbents and storing it at low pressure. Traditional natural gas storage tanks are thick and heavy, which makes them expensive to manufacture and highly energy-consuming to carry around. Our design uses unique adsorbent pellets with nanoscale pores surrounded by a coating that functions as a valve to help manage the pressure of the gas and facilitate more efficient storage and transportation. We expect this new concept will result in a lighter, more affordable product with increased storage capacity. The nanovalved adsorbent concept demonstrated here can be potentially extended for the storage of other important gas molecules targeted for diverse relevant functional applications.

Morphologic severity of cirrhosis determines the extent of liver resection in patients with hepatocellular carcinoma and Child-Pugh grade A cirrhosis.

BACKGROUND: Liver resection is the mainstay of treatment for patients with hepatocellular carcinoma and compensated cirrhosis. We investigated the relationship between the morphologic severity of cirrhosis and post-hepatectomy liver failure (PHLF) and evaluated the role of cirrhosis staging in determination of the extent limit for liver resection. METHODS: The clinicopathologic data of 672 consecutive patients with Child-Pugh grade A liver function who underwent curative liver resection for hepatocellular carcinoma in Tongji Hospital from 2009 to 2013 were retrospectively reviewed. Severity of cirrhosis was staged morphologically and histologically. Risk factors for histologic cirrhosis and PHLF were analyzed. The extent limit of liver resection with reference to morphologic staging was studied. RESULTS: Morphologic and histologic stages were significantly correlated (τ = 0.809, P < 0.001). Multivariate analysis showed that morphologic staging was the most crucial factor for histologic cirrhosis (odds ratio = 26.99, 95% confidence interval = 16.88-43.14, P < 0.001) and PHLF (odds ratio = 11.48, 95% confidence interval = 6.04-21.82, P < 0.001). The incidence of PHLF was high in patients with mild cirrhosis after resection of four or more liver segments (13.6%), those with moderate cirrhosis after major resection (38.1%), and those with severe cirrhosis or severe portal hypertension after resection of two or more liver segments (63.2% and 50.0%, respectively). CONCLUSIONS: Morphologic severity of cirrhosis is an independent predictor of PHLF. Resection of fewer than four liver segments is justified in patients with mild cirrhosis. Major resection is not recommended in patients with moderate cirrhosis. In patients with severe cirrhosis or severe portal hypertension, only resection of fewer than two liver segments can be safely performed.

Primary results of salvage liver transplantation in the patients with unresectable recurrent hepatocellular carcinoma after initial liver resection.

BACKGROUND/AIMS: Salvage liver transplantation (SLT) is a treatment choice for recurrent HCC fulfilling the Milan criteria. However, there is no consensus on the value of SLT for recurrent HCC beyond the Milan criteria, especially for unresectable HCC. METHODOLOGY: Eleven patients with recurrent HCC underwent SLT in Tongji Hospital between January 2003 and July 2010. All the 11 patients were considered unresectable because of deteriorated liver function, multiple bilobar tumors or vascular invasion. The outcomes and prognostic factors of these patients were analyzed. RESULTS: At a median follow up of 30 months, six patients were alive. Four patients died from HCC recurrence, and one died from gastric cancer. The 1-, 2-, and 3- year recurrence and overall survival rates after SLT were 58.4%, 72.3% and 86.1%, respectively, and 90.9%, 40.6% and 40.6%, respectively. Vascular invasion, recurrent HCC beyond the Milan criteria and early recurrence within 18 months after initial resection were negative prognostic factors of SLT for recurrent HCC. CONCLUSIONS: SLT can be recommended as an alternative treatment for recurrent HCC fulfilling the Milan criteria. For those beyond the Milan criteria or with vascular invasion, or early recurrence after initial resection, however, SLT is not beneficial and should not be recommended.

Decreased expression of serum semaphorin 3B is associated with poor prognosis of patients with hepatocellular carcinoma.

Semaphorin 3B (SEMA-3B), which belongs to the semaphorin family, has an important role in cell apoptosis and inhibition of angiogenesis. A previous study by our group revealed that SEMA-3B was downregulated in tumor tissues of patients with hepatocellular carcinoma (HCC) and exerts anti-motility and anti-invasion effects on tumor cells. However, the serum levels of SEMA-3B and their clinical significance have remained elusive; therefore, the aim of the present study was to monitor its expression in HCC and investigate its clinical significance. ELISA was used to determine the serum levels of SEMA-3B in 132 patients with HCC and 57 healthy individuals. The association between SEMA-3B and clinicopathological parameters was investigated. Serum SEMA-3B was indicated to be significantly decreased in patients with HCC as compared with that in the controls (P<0.05) and it was negatively associated with tumor size (P=0.039), encapsulation (P=0.002) and TNM stage (P=0.034). The prognosis of patients with low expression of SEMA-3B was poor. In conclusion, the results of the present study revealed that serum SEMA-3B is decreased in HCC and is negatively associated with prognosis; therefore, it may be used as a prognostic marker in HCC.

Role of PEPT1in the transport of cinnabar in Caco-2 cells.

Cinnabar, a mercury-containing mineral medicine, has been used as an ingredient in Traditional Chinese Medicines for treatment of various diseases for thousands of years and is still widely used today. The toxicity of cinnabar is much less than other mercury-containing compounds. This study aimed to evaluate the possible role of oligopeptide transporter1 (PEPT1) in intestinal uptake of cinnabar. Thus, the Caco-2 cell model was employed to investigate the differential transport levels and the probable transporter involved in the transport of cinnabar, mercury sulfide (HgS) and mercury chloride (HgCl2). Cells were incubated with the same molar concentration of cinnabar, HgS or HgCl2 and then the inorganic mercury content of apical (AP), cellular and basolateral (BL) side of the cell was measured by ultra-high liquid chromatography-inductively coupled plasma mass spectrometry (UPLC-ICP/MS) after the treatment, respectively. Their transportation levels were also investigated when pH was changed to 5.5 in AP side to define the role of the H+ dependent transporter. Effects of cinnabar, HgS or HgCl2 on transporter mRNA and protein expression levels were assayed by RT-PCR and Western-blot method, respectively. The possible transporter involved in the transport was examined by siRNA silencing and chemical inhibition. The results showed that the levels of inorganic mercury in the BL side for cinnabar and HgS were 49.39% and 30.41% of that in HgCl2 group. The transport levels of cinnabar and HgCl2 were significantly increased when the pH was changed to 5.5 on the AP side as compared with the control group (pH 7.4). Cinnabar significantly decreased the mRNA and protein expression of PEPT1. Transport levels of cinnabar were significantly decreased by PEPT1-siRNA and chemical inhibition of PEPT1. The present study demonstrates that PEPT1 may be an important transporter in the entry of cinnabar into the intestinal epithelium, and intestinal transport levels of cinnabar and HgS was lower than that of HgCl2.

CaCox Zr1-x O3-δ Perovskites as Oxygen-Selective Sorbents for Air Separation.

ABO3-δ perovskites are ideal for high-temperature thermochemical air separation for oxygen production because their oxygen nonstoichiometry δ can be varied in response to changes in temperature and oxygen partial pressure [ p O 2 ]. Herein, the outstanding oxygen-sorption performance of CaCox Zr1-x O3-δ perovskites and their potential application as oxygen-selective sorbents for air separation is reported. In situ thermal X-ray diffraction was used to study the materials' structural changes in response to temperature variations in air and inert atmosphere. Temperature-programmed reduction was employed to elucidate the relationship between perovskite composition and redox property. O2 sorption performance was evaluated by isothermal analyses at various temperature and p O 2 along with long-term absorption-desorption cycle tests. The high oxygen-sorption capacity was mainly attributed to Co at B-site, whereas partial substitution of Co by Zr enhanced the structural crystallinity and thermal stability of the perovskite. A stable oxygen production of 2.87 wt % was observed at 900 °C during 5 min-sorption cycles for 100 cycles.

Levels, congener profiles, and dietary intake assessment of polychlorinated dibenzo-p-dioxins/dibenzofurans and dioxin-like polychlorinated biphenyls in beef, freshwater fish, and pork marketed in Guangdong Province, China.

Persistent organic pollutants such as polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (DL-PCBs) consisting of non-ortho and mono-ortho PCBs are suggested to be very hazardous and have adverse effects on human health. However, their levels and congener profiles in retail foods marketed in Guangdong Province of China have not been elucidated thus far. Thus, in this study, 226 individual samples of beef, freshwater fish, and pork marketed across four regions of Guangdong Province were randomly collected during 2013-2015 to determine their levels of PCDD/Fs and DL-PCBs. The results showed that the total toxic equivalency quantities (TEQs) of most samples were below the maximum limits except for the 26 samples collected from the vicinities of pollution areas. The median total TEQs of these three categories were 0.174, 0.488, and 0.113pgTEQ/g fw, respectively, which indicated that the contamination status of the studied foods was not serious. For congener profiles, significantly different patterns were observed in three food groups, but with the same major TEQ contributors being 2,3,4,7,8-PeCDF in beef, freshwater fish, and pork. Regional differences of congener profiles in each food group were also found in this study, which might be attributed to the regionally different distributions of PCDD/Fs and DL-PCBs in environment media. The dietary exposures of four population subgroups (girls, boys, male adults, and female adults) to PCDD/Fs and DL-PCBs via three food groups were estimated to assessed the potential risks. They were all lower than the provisional tolerable monthly intake (PTMI, 70pgTEQ/kgbw/month) established by Joint FAO/WHO Expert Committee on Food Additive. In these food categories, the exposure to PCDD/Fs and DL-PCBs via freshwater fish was the highest one, which accounted for about 20% of PTMI, indicating that it was the major route to expose dioxin compounds.

Pancreatogenic choledocholithiasis in common bile duct stump after Roux-en-Y hepaticojejunostomy: A case report.

RATIONALE: Choledocholithiasis in common bile duct (CBD) stump after Roux-en-Y hepaticojejunostomy (RYHJ) is incredibly rare and its pathophysiology is poorly understood. PATIENT CONCERNS: A 79-year-old woman was admitted to our hospital with upper abdominal pain radiating through to the back in November 2016. DIAGNOSES: Abdominal computed tomography (CT) scan and magnetic resonance cholangiopancreatography (MRCP) revealed filling defects in CBD stump, chronic pancreatitis, and dilatation of CBD stump and main pancreatic duct (MPD). INTERVENTIONS: During the endoscopic retrograde cholangiopancreatography (ERCP), cannulation proceeded easily from MPD to CBD through a variant pancreatic duct, and then white crushed stones extracted from the CBD stump. Elemental analysis and infrared spectrophotometry demonstrated that the main constituent of the calculi was calcium carbonate. OUTCOMES: After a therapeutic ERCP, the patient's symptoms disappeared, and a 9-month follow-up indicated no remaining stones or lithiasis relapse. LESSONS: This type of choledocholithiasis in CBD stump after RYHJ has never been reported before. We nominated it as "pancreatogenic choledocholithiasis," and pancreatobiliary reflux caused by a variant pancreatic duct may be the main cause.

Hepatitis B virus X protein promotes CREB-mediated activation of miR-3188 and Notch signaling in hepatocellular carcinoma.

Familiar clustering of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) has been frequently reported. However, limited information is available about the underlying molecular mechanisms in HBV-related HCC patients with family history of HCC. In our previous study, Agilent miRNA Base 16.0 microarray showed miRNA profiles of the plasma of HBV-related HCC patients who had a family history of HCC. This study aims to explore the expression, function, and mechanisms of miR-3188 in HCC that might provide novel insights into the role of family history on the risk of HCC. The expression levels of miR-3188 were markedly overexpressed in HCC tissues, HBV transgenic mice, and HepG2.215 cells. We knocked out miR-3188 in HCC cell lines using the CRISPR/Cas9 system, and demonstrated that miR-3188 knockout (KO) suppressed cell growth, migration, and invasion, and inhibited xenografts tumor growth in nude mice. Next, we determined that miR-3188 KO exerts antitumor functions by directly repressing ZHX2. It has been reported that HBV X protein (HBx) plays a critical role in HBV-related HCC, promoting CREB-mediated activation of miR-3188 and activation of Notch signaling through repressing ZHX2. Finally, we verified that ZHX2 functions as a transcriptional repressor to Notch1 via interaction with NF-YA. Our data demonstrate that the HBx-miR-3188-ZHX2-Notch1 signaling pathway plays an important role in the pathogenesis and progression of HBV-related HCC with family history of HCC. These findings have important implications for identifying new therapeutic targets in HBV-related HCC.

Distilled Water Lavage During Surgery Improves Long-Term Outcomes of Patients with Ruptured Hepatocellular Carcinoma.

BACKGROUND: Rupture of hepatocellular carcinoma (HCC) releases tumor cells and furthers peritoneal metastasis. The present study investigated the killing effects of distilled water (DW) on HCC cells and the clinical outcomes of patients undergoing liver resection with DW lavage for spontaneously ruptured HCC. MATERIALS AND METHODS: Human HCC cells (BEL-7402, SMMC7721) were treated with DW, the morphological changes observed, and cell viability measured. DW-treated HCC cells were also injected intraperitoneally into nude mice, and the formation of tumor nodules and overall survival (OS) measured. The clinicopathological data of 141 consecutive patients undergoing liver resection for spontaneously ruptured HCC during 1998-2011 were retrospectively reviewed. RESULTS: Fifteen minutes of DW exposure caused complete cell lysis of HCC cells in vitro and completely prevented tumor formation and prolonged survival time in nude mice. Among the 141 patients, the 1-, 3-, and 5-year disease-free survival (DFS) and OS rates in patients administered DW lavage during surgery were 68.9, 24.6, and 6.6%, respectively, and 95.1, 65.1, and 40%, respectively, which were significantly higher than those in patients who did not (P < 0.05). DW lavage was an independent predictor of recurrence (odds ratio (OR), 0.34; 95% confidence interval (CI), 0.23-0.51; P < 0.001) and OS (OR, 0.35; 95% CI, 0.23-0.53; P < 0.001). CONCLUSIONS: Fifteen minutes of DW lavage can effectively kill HCC cells in vitro and prevent tumor formation in vivo. DW lavage significantly improves long-term outcomes in patients undergoing liver resection for spontaneously ruptured HCC and could be administered intra-operatively when tumor cell liberation is suspected.

Minocycline and cisplatin exert synergistic growth suppression on hepatocellular carcinoma by inducing S phase arrest and apoptosis.

Minocycline, a semisynthetic tetracycline, is a highly lipophilic molecule capable of infiltrating tissues and blood. Previous studies have revealed the functions and mechanisms of minocycline in anti-inflammation, protection of the nervous system and certain tumors. The role of minocycline has never been investigated in hepatocellular carcinoma (HCC). The functions of minocycline on HCC cells were investigated using immunohistochemical staining and western blotting. Minocycline was applied to L02, HepG2 and Huh7 cells, and the growth characteristics were studied. Cisplatin was administered in combination with minocycline in this study. Cell cycle and apoptosis analyses were employed to investigate the mechanisms underlying the growth regulation associated with minocycline and(or) cisplatin. Minocycline caused S phase cell cycle arrest and an increase in the apoptotic rate associated with upregulation of p27, cleaved-caspase8, cleaved-caspase3 and cleaved-PRAP-1. Low dose of cisplatin promoted cell cycle arrest and apoptosis, whereas minocycline was mainly associated with upregulation of cleaved-PARP-1. The combination of cisplatin and minocycline increased the rate and extent of cell cycle arrest and increased the apoptosis rate caused by minocycline. A novel mechanism was revealed. Minocycline functions as an antitumor drug in HCC by regulating p27, caspase-3 and PARP-1. Cisplatin enhanced minocycline's effect on PARP-1.

[The change of macular thickness of SAS before and after sleep].

PURPOSE: To observe the changes of macular thickness in patients with sleep apnea syndrome (SAS) before and after sleep. METHODS: Thirty-two patients (63 eyes) diagnosed as SAS from August 2003 to January 2007 were enrolled. Macular thickness were measured using optical coherence tomography (OCT) at 11:00 to 12:00 evening (before sleep)and 20 to 30 minute after sleep, respectively. RESULTS: The mean macular thickness was (123.00+/-19.98)um and (134.25+/-19.92)um before and after sleep, respectively. The mean difference was (11.25+/-9.04)um before and after sleep(t=9.878,p<0.05),95%CI(8.98,13.53)um. CONCLUSIONS: The macular thickness of SAS is increased in SAS patients, which may due to anoxia of SAS.