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Sorafenib is the oldest first line systemic treatment in patients with advanced hepatocellularcarcinoma (HCC) and has been used exclusively for nearly 10 years. The superiority ofadministering a combination of atezolizumab plus bevacizumab (AteBeva) compared tosorafenib as first line systemic treatment for unresectable HCC was recently proven duringthe IMbrave150 Phase III randomized trial. While clinicians can expect improved responsesand treatment outcomes due to the good results of the IMbrave 150 trial, they must alsoconsider that atezolizumab can cause various immune-related adverse events (IrAEs). Basedon the above suggestions, we herein present a case of HCC with lymph node metastasiswho achieved complete remission following treatment with AteBeva and developed an IrAE(adrenal insufficiency). Further study of real-life data regarding combination therapy withAteBeva is needed to manage patients with advanced HCC.

Lipases catalyze a wide range of industrially important reactions, including the transesterification of triglycerides with alcohols for biodiesel production, and the stabilization of lipases are critical to achieve their recycled uses. Here, nanoscale enzyme reactor (NER) of lipase from Rhizopus oryzae (LP) was prepared via a simple two-step process, comprising of enzyme adsorption into magnetically-separable mesoporous silica and follow-up crosslinking of adsorbed enzymes. In aqueous phase, the specific hydrolysis activity of NER-LP was 4.7 times lower than that of free LP. On the other hand, however, the specific transesterification activity of NER-LP (130.4 μmol/min/mg LP) in organic phase for biodiesel production was 50 times higher than that of free LP (2.6 μmol/min/mg LP). These results reveal that the enzyme crosslinking for the preparation of NER does not interfere with the interfacial activation of LP molecules, opening the lid of LP active site under an optimal hydrophobic environment provided by the combination of organic solvent and mesoporous silica. Magnetic separation and optimized washing protocol facilitated the recycled uses of NER-LP. Highly stable and active NER-LP in magnetically-separable mesoporous silica has demonstrated its great potentials as an environmentally-friendly nanobiocatalyst for various lipase applications, including plasticizers, biosurfactants, functional fatty acids, as well as recyclable biodiesel production.

Background: Hand-foot skin reaction (HFSR) is the most common adverse event during sorafenib treatment, which is the main reason for dose de-escalation or discontinuation of sorafenib in hepatocellular carcinoma (HCC) patients. Here, we aimed to investigate the role of urea cream in the prevention or amelioration of HFSR severity. Methods: HCC patients at 13 hospitals in Korea were recruited from May 2016 to May 2018. Patients were treated with placebo cream and urea cream for 12 weeks concomitantly with sorafenib treatment. Development of HFSR, score for Hand-Foot Skin Reaction and Quality of Life (HF-QoL) questionnaire, and adverse event were assessed at 2, 4, 8, and 12 weeks. Findings: Of the 288 patients, 247 patients, with 117 patients in the placebo control group and 130 patients in the urea cream group, were analyzed. The urea cream group showed lower cumulative incidence of any grade of HFSR (Log-rank, p=0.247) and severe HFSR of grade 2 or higher (Log-rank, p = 0.394) without statistical significance. In the incidence by time point, the development of severe HFSR of grade 2 or higher was significantly lower in the urea cream group than in the placebo control group at two weeks (13.8% vs. 23.9%, p=0.042). The urea cream group showed a significantly improved questionnaire score compared to the placebo control group (11.8 vs. 19.7, p=0.014) at 12 weeks. Interpretation: Treatment with urea cream showed a lower incidence of severe sorafenib-induced HFSR at 2 weeks and reduced the tendency of HFSR development in HCC patients. Therefore, treatment with urea cream could be considered for prophylaxis or improvement of HFSR grade in HCC patients treated with sorafenib. Trial Registration: This study was registered at ClinicalTrial.gov (NCT03212625). Funding Statement: The Korea Liver Cancer Association and Hanmi Pharmacy (IIT-CAV-004). Declaration of Interests: The authors declare no competing interests. Ethics Approval Statement: All participants provided written informed consent. This study was approved by the institutional review board of each participating institute (2016GR0350).

Sorafenib is a well-known approved systemic therapeutic agent used in patients with advanced hepatocellular carcinoma (HCC). Regorafenib and nivolumab are approved as second-line therapeutic drugs in patients showing disease progression after sorafenib therapy. However, there is no established third- or fourth-line therapy in patients with progression after regorafenib or nivolumab treatment. Recently, the combination of tyrosine kinase inhibitors (TKIs) and immune checkpoint inhibitors (ICPIs) has been attempted as a firstline treatment strategy in advanced HCC patients based on the hypothesis that combination therapy may overcome resistance in ICPI monotherapy. On the basis of this suggestion, we herein describe the case of an HCC patient demonstrating macrovascular invasion, whereby partial remission was achieved via the combination of sorafenib and nivolumab following disease progression after nivolumab therapy. Further studies on the combination of TKIs and ICPIs are necessary to determine ways to manage HCC patients showing disease progression after ICPI therapy. (J Liver Cancer 2019;19:74-78)

Background/Aims Regorafenib has been approved as a second-line systemic therapy for hepatocellular carcinoma (HCC) patients after the phase III RESORCE trial. This study analyzed real-world data to assess the clinical effectiveness and safety of regorafenib compared to the RESORCE trial. Methods This multicenter cohort study included HCC patients treated with regorafenib after sorafenib (n = 133). We evaluated the time to progression (TTP), progression-free survival (PFS), overall survival (OS), and safety in patients receiving regorafenib along with the predictors of prognosis. Results The median age was 60 years and 81.2% patients were men. Hepatitis B virus infection (68.4%) was the commonest etiology. Most patients were classified as Child-Pugh A (98.5%) and had extrahepatic metastasis (84%) and vascular invasion (45.1%). This study demonstrated similar characteristics apart from more frequent hepatitis B etiology and more vascular or extrahepatic involvement compared with the RESORCE trial. An objective response rate of 12.5% was obtained for response assessment (n = 112); the disease control rate was 34.8%. Thirty-eight patients died during follow-up. With regorafenib, the median OS, PFS, and TTP were 10.0, 2.7, and 2.6 months, respectively. In the exploratory analysis after sorafenib administration, the median OS was 25.8 months. The rate of response and survival were comparable to those in the RESORCE trial. Child-Pugh score > 5, alpha-fetoprotein > 400 ng/ml, and TTP for sorafenib ≥ median were independently associated with OS. Conclusions This real-word regorafenib study showed comparable effectiveness and safety to the RESORCE trial. Regorafenib improves the prognosis of patients with prolonged TTP during previous sorafenib therapy.

Amylases from Streptomyces are useful in the production of maltooligosaccharides, but they have weak thermal stability at temperatures higher than 40 °C. In this study, α-amylase (SAV5981 gene of Streptomyces avermitilis) was expressed from Streptomyces lividans 1326 and purified by ammonium sulfate fractionation followed by anionic chromatography (Q-HP sepharose). The properties of the purified SAV5981 amylase were determined by the starch-iodine method. The effect of metal ions on amylase activity was investigated. The optimal temperature shifted from 25 to 50 °C with the addition of the Ca(2+) ion. The thermal stability of SAV5981 was also dramatically enhanced by the addition of 10 mM CaCl2. Improvement of the thermal stability of SAV5981 was examined by CD spectra in the presence and the absence of the Ca(2+) ion. Thin-layer chromatography (TLC) analysis and HPLC analysis of starch degradation revealed that SAV5981 mainly produced maltose and maltotriose, not glucose. The maltoorigosaccharide-producing amylase examined in this study has the potential in the industrial application of oligosaccharide production.

Encapsulation of active proteins in the hydrophilic core of vesicular liposomes is important for developing a therapeutic protein carrier system. The efficiency of liposomal encapsulation of proteins is generally low. A better understanding of the fundamental mechanisms of encapsulation is needed to increase this efficiency. In this study we investigated the effects of operating parameters such as phospholipid concentration, buffer pH and ionic strength, protein size and surface charge, and liposome size on the enzyme encapsulation yield. Four model enzymes of different molecular weights and isoelectric points (trypsin, horseradish peroxidase, enterokinase and hyaluronidase) were encapsulated into three different sized liposomes (125 nm, 194 nm, and 314 nm in mean diameter). Relatively inert and neutral DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphocholine) was used as the main phospholipid in the liposomes. Size exclusion chromatography was used to separate the enzyme-encapsulated liposomes from the free enzyme, and the encapsulation yield was determined from the peak area. The encapsulation yield was generally low ranging from ca. 5% to 20%, and did not depend much on the molecular weight of the enzyme encapsulated. Larger liposomes had higher encapsulation yields. The electrostatic interaction between the phospholipid and enzyme was the most significant parameter in determining the encapsulation yield. Thus adjusting buffer pH and ionic strength and adding charged phospholipids to the liposome preparation to impart electric charge to the lipid bilayer could significantly improve the yield. This approach can be used to optimize the liposomal encapsulation of clinically significant proteins.

The objective of this study was to propose a feasibility of a cellular imaging assay as an alternative to the conventional cytotoxicity assay, such as MTS assay, for apoptosis monitoring. As an apoptosis monitoring parameter, affinity interaction between phosphatidylserine (PS) and annexin V was chosen. First, the specific binding affinity between annexin V and PS in phospholipid bilayers consisting of various molar (0-15%) composition of PS was measured using a surface plasmon resonance biosensor. As PS composition increased, the binding level of annexin V increased proportionally. Second, various concentrations (0.1-10 μM) of staurosporine were used as to induce apoptosis and introduced to MCF-7 breast carcinoma cells. The cellular fluorescence images from annexin V-FITC conjugate were obtained by confocal microscopy, and their fluorescence intensities were quantified by image scanning. Dose-apoptosis (or cell death) relationships were very similar to those from MTS and FACS assays. In summary, our cellular imaging method could serve as a quicker and simpler alternative to MTS (end point assay) and FACS (flow cytometry) to screen potential apoptosis inducers.

생촉매인 효소의 기질선택성은 다양한 분야에서 유용하게 이용되고 있다. 그 중에서도 효소결합면역흡착검사(enzyme-linked immunosorbent assay, ELISA)는 항원항체의 결합을 항체와 공유결합된 효소의 반응으로 나타냄으로써 다양한 항원들의 진단을 가능케 했다. 하지만 기존의 효소결합면역흡착검사는 하나의 항체당 하나의 효소가 결합된 형태이기 때문에 감도(sensitivity)의 증가 폭에 그 한계가 있으며, 이를 극복하기 위한 방안으로 하나의 항체당 결합된 효소의 수를 증가시킴으로써 혁신적인 감도의 향상을 가져오는 연구가 진행되었다. 최근 나노바이오촉매(nanobiocatalyst, NBC) 접근방식을 이용한 효소활성의 안정화는 효소결합면역흡착검사의 감도 향상뿐만 아니라 그 성능의 안정성을 확보할 수 있는 연구결과로 이어지고 있다. 본 총설에서는 일반적인 효소결합면역흡착검사의 기본적인 원리와 감도향상을 위한 연구, 그리고 성능안정성(performance stability)을 향상시키기 위한 나노바이오촉매-결합면역흡착검사(Nanobiocatalyst-Linked Immunosorbent Assay, NBC-LISA)에 대하여 살펴보고자 한다. 【Enzymes are being used in various fields due to their unique property of substrate specificity. Enzyme-linked immunosorbent assay(ELISA) has enabled the detection of various antigens by reporting the binding event of antigen and antibody via enzyme-catalyzed reaction. However, the sensitivity improvement of conventional ELISA has been limited because only one enzyme molecule is conjugated to one molecule of antibody. To overcome this limitation and further improve the sensitivity of ELISA, there have been efforts to increase the number ratio of enzymes to antibody. Recently, the nanobiocatalytic approaches, with their successful enzyme stabilization, improved the performance stability as well as sensitivity in a modified protocol of ELISA. The present paper introduces the basic principle of ELISA, and the recent efforts to improve sensitivity and performance stability of ELISA by using the nanobiocatalytic approaches.】

Caspase-3 is a key mediator of apoptosis in mammalian cells. Cells expressing caspase-3 substrate peptides have become powerful and increasingly common components of cell-based assay systems. We developed a cell-based assay to measure staurosporine (STP)-induced caspase-3 activity in live rat glioma C6 cells. The caspase-3 sensing system was constructed to include a nuclear export signal (NES), followed by the amino acid sequence Asp-Glu-Val-Asp (DEVD) and a green fluorescent protein (GFP) fused to the Nterminal site of a nuclear localization signal (NLS). Using time-lapse confocal microscopy, we monitored caspase-3 activation during apoptosis by imaging the translocation of GFP from the cytosol to the nucleus. After 8 h of 0.5 μM STP treatment, caspase-3 activity was assessed by monitoring the translocation of GFP to the nucleus due to cleavage of the NES from the GFP by caspase-3. Finally, disintegration of the plasma membrane during late apoptosis was confirmed using a nuclear dye, propidium iodide. Analysis of caspase-3 activity using real-time monitoring can potentially be used to screen for apoptotic molecules in living cells.

Lipase (LP) was immobilized on electrospun and ethanol-dispersed polystyrene-poly(styrene-co-maleic anhydride) (PS-PSMA) nanofibers (EtOH-NF) in the form of enzyme precipitate coatings (EPCs). LP precipitate coatings (EPCs-LP) were prepared in a three-step process, consisting of covalent attachment, LP precipitation, and crosslinking of precipitated LPs onto the covalently attached LPs via glutaraldehyde treatment. The LP precipitation was performed by adding various concentrations of ammonium sulfate (20-50%, w/v). EPCs-LP improved the LP activity and stability when compared to covalently attached LPs (CA-LP) and the enzyme coatings of LPs (EC-LP) without the LP precipitation. For example, the use of 40% (w/v) ammonium sulfate resulted in EPC40-LP with the highest activity, which was 4.0 and 3.6 times higher than those of CA-LP and EC-LP, respectively. After 165-day incubation under rigorous shaking at 200 rpm, the residual activities of EPC50-LP were 0.5 μM/min mg of EtOH-NF, representing 113 and 75 times higher than those of CA-LP and EC-LP, respectively. When LP was partially purified via a simple ammonium sulfate precipitation and dialysis, both activities and stabilities of EC-LP and EPC-LP could be marginally improved. It is anticipated that the improved LP activity and stability in the form of EPCs would allow for their potential applications in various bioconversion processes such as biodiesel production and ibuprofen resolution.

Goal: In this study, we attempted to evaluate the prognosis of combined hepatocellular and cholangiocarcinoma (cHCC-CC) with comparison to hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (CC). Background: The prognosis of cHCC-CC has not been fully elucidated. In this study, we attempted to evaluate the prognosis of cHCC-CC with comparison to HCC and CC. Study: Consecutive patients who underwent curative resection for cHCC-CC at a single tertiary care center in Korea and their age, sex, and Child-Turcotte-Pugh class matched HCC and CC patients were included. We evaluated time-to-recurrence (TTR) and overall survival (OS) of cHCC-CC cases and compared them with HCC and CC patients. Results: Thirty cHCC-CC, 60 HCC, and 60 CC patients were included. For cHCC-CC group, the median TTR and OS were 5.4 and 18.0 months. After adjustment for confounding factors, the cHCC-CC group had a shorter TTR than did HCC group [relative risk (RR), 2.76; P < 0.001] and CC group (RR, 2.00; P = 0.013), and a shorter OS than HCC group (RR, 4.70; P < 0.001). Compared with the each stage I diseases, cHCC-CC had shorter TTR than HCC (RR, 4.59; P = 0.001) and CC (RR, 2.74, P = 0.015) and shorter OS than HCC (RR, 9.35; P = 0.001). Conclusions: The results of this study indicated that cHCC-CC had a significantly poorer prognosis than HCC and CC even after curative resection.

To investigate the effect of cholesterol composition on the binding of factor VIII (FVIII) and annexin V (AV) to membranes, liposomal membranes with phospholipid bilayers of various compositions of phosphatidylcholine (PC), phosphatidylserine (PS), and cholesterol were constructed. A surface plasmon resonance (SPR) biosensor system was employed to measure the equilibrium and rate constants of the bindings. As expected, PS was found to play a dominant role in the binding of AV; its binding level was directly proportional to the PS composition in a liposome. The binding levels of FVIII and AV to liposome increased with an increase in cholesterol composition in liposome. It seemed to suggest that cholesterol in liposome acts as a 'phospholipid arrangement' factor by inducing the formation of PS-rich microdomains. However, in the absence of PS (20% on a mole basis), cholesterol could not exert the binding enhancement effect, which again confirmed the critical role of PS in the bindings. Stability of the AV binding was significantly improved by the increase in cholesterol content; for AV, the dissociation rate constant was decreased approximately fivefold, from 1.7 x 10(-3)s(-1) in the absence of cholesterol to 3.3 x 10(-4)s(-1) in the presence of only 10% cholesterol. But, for FVIII the binding stability was not so much influenced by the cholesterol addition (up to 50% on a mole basis). In summary, by using liposomes on an SPR system, we were able to demonstrate quantitatively the apparent effects of cholesterol on the binding affinity and stability of the membrane-binding proteins.

SiO(2) surface was successfully modified with phospholipid bilayer for biocompatibility by covering the planar surface with vesicular liposomes. By applying heat to rupture the vesicle, they were converted into a planar form. To effectively decorate the bilayer with biological molecules such as a protein, BAM (biological anchor for membrane) was used as a linker. It is a linear assembly consisting of oleyl chain, polyethylene glycol, and NHS (N-hydroxysuccinimide). After a target protein (BSA) was conjugated with BAM by NHS replacement, the conjugate was effectively inserted to the phospholipid bilayer through the lipophilic interaction between the oleyl chain and the lipid bilayer. The entire process was monitored and quantitatively analyzed by QCM (quartz crystal microbalance). BSA-BAM conjugate showed approximately 12-fold higher binding efficiency to the lipid bilayer than BSA alone. From this result, we conclude that SiO(2) surface could be modified to a lipid bilayer surface so as to anchor a protein by the action of BAM.

Liposome immunoassay (LIA) is based on enzyme immunoassay (EIA) but the detection sensitivity could be significantly enhanced by using antibody-coupled immunoliposomes encapsulating HRP (horse radish peroxidase). Here, we applied LIA to non-porous poly(methyl methacrylate) (PMMA) and polystyrene (PS) surfaces to compare its detection sensitivity with that of EIA, using rabbit IgG (a ligand molecule) and anti-rabbit IgG antibody (a capture molecule) as the model system. LIA developed much stronger color signals than EIA, especially at a lower concentration range (ca. 1 microg mL(-1)). PMMA showed higher affinity toward rabbit IgG than the PS surface, and the anti-rabbit IgG antibody adsorbed on PMMA was more stable than that on PS. Furthermore, the effects of spot volume and antibody concentration on the signal density were analyzed. The signal density increased as the antibody concentration increased, but it was not significantly affected by the spot volume (2.5-20 microL). In conclusion, LIA on PMMA as a solid support is a very useful, highly sensitive microarray detection system.

Recent technical advances in biorecognition engineering and microparticle fabrication enabled us to develop a single-step purification process using magnetic particles (MPs). The process is simple, efficacious, easy to automate, and economical. The method immobilizes the ligand molecule in a particular orientation on commercial MPs that have surface carboxyl groups. Mouse IgG and anti-mouse IgG antibody were the model capture and ligand molecules for this study. The immunobinding efficacy of anti-mouse IgG antibody using “oriented immobilization” was compared with the efficacy of a conventional amine-coupling system that results in random orientation and of another standard method, the biotin–streptavidin system. The oriented immobilization was accomplished by oxidizing the sugar moiety in the CH 2 domain of the antibody's Fc and covalently conjugating the moiety to the hydrazine-coated MP. The specific binding affinity of the oriented immobilization process was about 2.5 times that of the amine-coupling system, and selectivity from a binary mixture was about 2 times greater for the oriented immobilization method. Results were nearly identical for the biotin–streptavidin system and the oriented immobilization system, matching the calculated binding stoichiometry between mouse IgG and anti-mouse IgG antibody. The binding improvement over the amine-coupling system shown by assay was confirmed by a separate surface plasmon resonance experiment. In summary, the oriented immobilization method was as effective as the streptavidin–biotin system, yet simpler and cost-effective.

Plant cells are increasingly used as alternative host organisms for recombinant protein production requiring post-translational modifications because they have relatively low risks of hazardous viral contamination compared to animal cells. On the other hand, the plant cell culture route has intrinsic disadvantages, including a relatively low expression titer and difficulties in the purification. We developed a simple and scalable, chromatography based process to purify rhGM-CSF from recombinant rice cell culture. It consisted of cell removal by microfiltration, ammonium sulfate precipitation, anion exchange chromatography, and SEC-HPLC. α-Amylase, used as a leader sequence in the expression vector, was the major impurity protein co-purified with rhGM-CSF. The purification process resulted in a final purity of ca. 95 % (as verified by size exclusion chromatography) and an overall recovery yield of ca. 48 % from the culture broth (as verified by ELISA).

The LAL (Limulus amebocyte lysate) test for the detection and quantification of endotoxin is based on the gelation reaction between endotoxin and LAL from a blood extract ofLimulus polyphemus. The test is labor intensive, requiring dedicated personnel, a relatively long reaction time (approximately 1 h), relatively large volumes of samples and reagents and the detection of the end-point is rather subjective. To solve these problems, a miniaturized LOC (labon-a-chip) prototype, 62 mm (L)×18 mm (W), was fabricated using PDMS (polydimethylsiloxane) bonded to glass. Using this prototype, in which 2 mm (W)×44.3 mm (L)×100 μm(D) microfluidic channel was constructed, turbidometric and chromogenic assay detection methods were compared, and the chromogenic method was found the most suitable for a small volume assay. In this assay, the kinetic-point method was more accurate than the end-point method. The PDMS chip chickness was found to be minimized to around 2 mm to allow sufficient light transmittance, which necessitated the use of a glass slide bonding for chip rigidity. Due to this miniaturization, the test time was reduced from 1 h to less than 10 min, and the sample volume could be reduced from 100 toca. 4.4 μL. In summation, this study suggested that the LOC using the LAL test principle could be an alternative as a semi-automated and reliable method for the detection of endotoxin.

Radioresistance in the tumor and radiotoxicity in the non‑tumorous liver significantly restrict efficient radiotherapy of hepatocellular carcinoma (HCC). It is therefore important to study the radioresistance mechanism and development of radiosensitization to optimize the effect of irradiation on cancer cells. Emodin (1, 3, 8‑trihydroxy‑6‑methylanthraquinone) is a plant‑derived polyphenol, possessing anticancer properties. It is known to act as a radiosensitizer in human HCC cell lines. The aim of this study was to evaluate the role of emodin in radioresistance of human HCC cell lines as well as the underlying radiosensitization mechanism. The human HCC cell line (HepG2) was used in this study. Four different treatment groups, i.e., no treatment (control), irradiation (10 Gy, one fraction), emodin (10 µM), and a combination of irradiation and emodin (10 Gy+10 µM) were used for two environmental conditions: hypoxia (1% O2) and normoxia (20% O2). The cells were exposed to the respective treatments for 24 and 72 h. Following the treatment, the cell viability was determined by the 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide (MTT) assay, and the radiosensitization mechanism was evaluated by western blotting. The proliferation of HepG2 cells was significantly suppressed in the treatment groups under hypoxic and normoxic conditions in the following order: combination of irradiation and emodin>irradiation only >emodin only. The combination of irradiation and emodin induced apoptotic signaling activities such as cleavage of poly (ADP‑ribose) polymerase (PARP)‑1 as well as the downregulation of epigenetic signaling such as JMJD1A and JMJD2B. Emodin attenuated radioresistance in the HepG2 cells via upregulation of the apoptotic signals and down-regulation of the proliferative signals. These results suggested that emodin is a potential candidate for the radiosensitization of HCC cells and can aid in identifying novel therapeutic strategies for HCC radiotherapy.

Treatment of cirrhotic patients with spontaneous peritonitis using antibiotics occasionally fails. Fungal infections may be one of the causes of antibiotic treatment failure in such patients. In this study we evaluated the clinical significance and characteristics of spontaneous fungal peritonitis (SFP). Consecutive cirrhotic patients with spontaneous peritonitis treated between 2000 and 2005 at a tertiary care center in Seoul, Korea, were included. We analyzed the clinical characteristics and the prognosis of SFP patients compared with patients with spontaneous bacterial peritonitis (SBP). During the study period 416 patients developed spontaneous peritonitis and 15 (3.6 %) had SFP. Compared with patients with SBP, nosocomial peritonitis (peritonitis that developed after hospitalization for >72 h) was more common and the Child–Pugh score was higher in SFP patients (both, P < 0.01). Ten patients were infected with Candida spp. (C. albicans, 8; C. tropicalis, 1; C. glabrata, 1), and 5 with Cryptococcus neoformans. Eleven patients were co-infected with bacteria that were susceptible to the antibiotics administered. Only 5 patients were treated using appropriate anti-fungal agents. The 1-month mortality rate for SFP patients was 73.3 % (11 out of 15; median time to death, 2 days [range, 0–22]), which was significantly higher than patients with SBP alone (28.7 %, P = 0.0007). SFP is severe complication related to high mortality in cirrhotic patients. A longer admission and a higher Child–Pugh score may be risk factors. Immediate anti-fungal treatment is warranted in patients with spontaneous peritonitis, once fungus is found in the ascitic fluid.

In this study, we aimed to devise a simple scoring system predicting the risk of genotypic resistance (GR) to current rescue therapies for patients with lamivudine (LAM)-resistant chronic hepatitis B.LAM and adefovir (ADV) combination therapy should be recommended for an initial rescue therapy against LAM-resistant hepatitis B virus (HBV). However, there still are many LAM-resistant patients being treated with entecavir (ETV) or ADV monotherapy.This retrospective cohort study included consecutive patients treated for LAM-resistant chronic hepatitis B with ETV or ADV monotherapy, or LAM/ADV combination therapy. The cumulative probabilities of GR and virological responses and breakthrough according to clinical variables were analyzed by survival analyses and derived an index for future GR.A total of 224 patients were included (median treatment duration=117.9 wk). Using risk factors indentified on multivariable analyses, a simple index for future GR (Antiviral Resistance Prediction Index, ARPI) was developed with 3 clinical variables: the rescue therapy regimens (+0, ADV; +1, ETV; +2, LAM/ADV), HBV DNA reduction at 12 weeks (+0,3 log10 copies/mL; +1,3 log10 copies/mL), and the initial HBV DNA level (+0,10 copies/mL; +1,10 copies/mL). No patient with ARPI ≥2 exhibited GR, whereas 47% of the patients with an ARPI2 developed GR by week 144 (P=0.005).The results of this study suggest that the ARPI is a simple and early index that can be used to determine the risk for subsequent GR during rescue therapy for LAM-resistant chronic hepatitis B.

To determine the efficacy and safety of transarterial chemoembolization (TACE) in patients with hepatocellular carcinoma (HCC) and main portal vein (MPV) invasion.This study was approved by the institutional review board, and the requirement to obtain informed consent was waived. The authors retrospectively assessed the electronic medical records of patients in whom HCC with MPV invasion was newly diagnosed from January 2004 to December 2007 at a single tertiary medical center. Patients with decompensated hepatic function were excluded. Outcomes of patients treated with TACE were compared with those of patients given supportive care according to Child-Pugh class.One hundred twenty-five patients (104 men and 21 women; mean age, 55.7 years; age range, 33.4-83.0 years) were included. The median overall survival was 3.7 months (range, 0.2-33.3 months). Eighty-three of the 125 patients (66.4%) were treated with TACE and 42 (33.6%) received supportive care. Repeated TACE showed significant survival benefits compared with supportive care in patients with Child-Pugh class A (median survival, 7.4 months vs 2.6 months, respectively; P.001) and class B (median survival, 2.8 months vs 1.9 months, respectively; P = .002) disease. Results of multivariate analysis showed that treatment with TACE (hazard ratio, 0.263; 95% confidence interval [CI]: 0.164, 0.424; P.001) and Child-Pugh class A status (hazard ratio, 0.550; 95% CI: 0.368, 0.822; P = .004) were independent predictive factors of a favorable outcome. There were no procedure-related deaths within 4 weeks after TACE, and patient morbidity was 28.9% (24 of 83 patients).TACE can be performed safely and may improve the overall survival of patients with HCC and MPV invasion.

Background/Aims Lamivudine (LAM) has been extensively used to treat hepatitis B, but high incidence of drug resistance has required rescue studies. We validated the optimum treatment strategy for LAM-resistant patients by means of a comparative study of add-on adefovir (ADV) and a switch to entecavir (ETV). Methods We assessed the virologic response in consecutive LAM-resistant patients who received add-on ADV or a switch to ETV. Results The mean reduction of serum hepatitis B virus (HBV) DNA levels was significantly less in the ETV group than in the add-on ADV group (-3.45 vs. -4.17; P = 0.047 at week 24 and -3.81 vs. -4.68 log10 IU/mL; P = 0.044 at week 48). Achievement of undetectable HBV DNA was significantly lower in the ETV group than in the add-on ADV group (P = 0.043). Multivariate analysis showed that add-on ADV, baseline HBV DNA levels, and initial virologic response were significant predictors of HBV DNA negativity (adjusted OR, 2.582; P = 0.008, 0.304; P = 0.001, and 5.928; P = 0.001). Virologic breakthrough was observed for 12 patients, in the ETV group only. Conclusions Add-on ADV was more effective and durable than ETV as rescue therapy. Therefore, add-on ADV might be the preferred strategy for LAM-resistant patients who need long-term antiviral treatment.

Xanthogranulomatous cholecystitis (XGC) is an unusual and destructive inflammatory process that is characterized by thickening of the gallbladder (GB) wall with a tendency to adhere to neighboring organs. XGC is often mis- taken for GB carcinoma, and the frequency of the coexistence of these two lesions is approximately 10%. Therefore, in case of severe XGC, there is chance of either overlooking the carcinoma or other significant lesions. CA 19-9 is commonly measured in the serum of patients with hepatobiliary malignancies. Although CA 19-9 can be elevated in benign conditions such as cholestasis, pancreatitis, tuberculosis, thyroid disease etc., ma- lignancy should be considered at first in setting of its significant and persistent elevation. We report a case of a 62-year-old man who showed continuously rising level of CA19-9 over 2000 U/mL after cholecystectomy for xanthogranulomatous cholecystitis and finally was diagnosed as cholangiocarcinoma by short-term follow up. (Korean J Gastroenterol 2010;55:404-409)

A novel HER2-targeted carrier was developed using bionanocapsules (BNCs). Bionanocapsules (BNCs) are 100-nm hollow nanoparticles composed of the l -protein of hepatitis B virus surface antigen. An affibody of HER2 was genetically displayed on the BNC surface (Z HER2 -BNC). For the investigation of binding affinity, Z HER2 -BNC was incubated with the cancer cell lines SK-BR-3 (HER2 positive), and MDA-MB-231 (HER2 negative). For analysis of HER2 targeting specificity, Z HER2 -BNC or Z WT -BNC (without affibody) was incubated with both SK-BR-3 and MDA-MB-231 cells by time lapse and concentration. For the delivery of encapsulated molecules (calcein), fluorescence of Z HER2 -BNC mixed with liposomes was also compared with that of Z WT -BNC and nude liposomes by incubation with SK-BR-3 cells. As a result, Z HER2 -BNC-liposome complex demonstrated the delivery to HER2-expressing cells (SK-BR-3) with a high degree of specificity. This indicates that genetically engineered BNCs are promising carrier for cancer treatment.

To develop a sterically stable, targeting proteoliposome, a postinsertion method was employed to biofunctionalize the liposome surface with a biocompatible anchor molecule (BAM) linker conjugated with epidermal growth factor (EGF) as a homing molecule. In this method, EGF was first conjugated with BAM that consisted of a hydrophilic reactive group at one end and oleic acid chains at the other end. The EGF-BAM complex was then inserted into the liposomal phospholipid bilayer through lipophilic interaction. When compared with the traditional surface modification method by amine coupling, the modification efficiency with BAM at 25 degrees C was about 2.5-fold higher, and the 24 h stability of the inserted BAM at 25 degrees C was also about 2.5-fold higher. The insertion affinity and stability of the BAM to the liposomal bilayer was not influenced by an increase in cholesterol concentration, a component in liposome preparation. Confocal microscopy studies showed that the proteoliposomes biofunctionalized with the BAM-EGF complex encapsulating Cy5 fluorescent dye were selectively bound to the surface of MDA-MB-231 cells overexpressing EGFR (epidermal growth factor receptor), but not to MCF-7 cells, which did not express EGFR. The same proteolipome encapsulating doxorubicin was selectively targeted to MDA-MB-231 cells and killed them. In sum, BAM could be used as a suitable postinsertion linker for biofunctionalization of liposome surface with high modification efficiency and stability. Furthermore, the protein used as a homing molecule maintained its bioactivity after the biofunctionalization.

One of the major problems of wild-type lignin peroxidase (LiP) is its inactivity at the presence of excess H(2)O(2) and high concentration of aromatic compounds. Little is known about the substrate-binding site of LiP, and functionality improvement of LiP was not actively tried by genetic engineering and directed evolution. In order to improve LiPs functionality, we performed directed evolution with a colorimetric screening method. Finally, three types of LiP mutants were screened. The catalytic efficiency of the variants toward 2,4-dichlorophenol (DCP) degradation activity and the stability against H(2)O(2) was increased over the wild type. The K(m) value of the variants toward H(2)O(2) was increased, but K(m) value toward 2,4-DCP degradation was reduced. Overall, The K(cat)/K(m) values of the mutants toward 2,4-DCP was increased ca. 4-fold, and that toward H(2)O(2) was increased ca. 89-fold. Amino acid sequence analysis indicated that the most of the mutations were located on the enzyme surface. We expect that these results coupled with recombining mutation can be successfully applied to the molecular evolution cycles for screening of LiPs and other oxidative enzymes with improved functionality and stability.

Rectal schwannoma is a rare mesenchymal tumor originating from Schwann's cell. We experienced a 61- year-old female patient who complained of blood tinged and narrow calibered stool for several years, and found a 4 cm sized submucosal tumor with a central ulcer on the rectal wall during colonoscopy. She underwent transanal excision. Microscopically, the tumor was composed of fasciculating bundles of spindle cells with benign nuclear atypia and peripheral lymphoid cell cuffing. Tumor cells showed a diffuse strong immunoreactivity to S-100 protein, but not stain for CD 34, desmin and smooth muscle actin. This is the first case report of rectal schwannoma in Korea.

Many patients suffering from breast carcinoma have metastases at initial diagnosis. The common metastatic sites are skeleton, liver and lung. Metastases to stomach are rare and only three cases have been reported in Korea. The endoscopic features of gastric metastases from breast carcinoma can be divided into three main categories: diffuse infiltration, external compression, and localized tumor deposition with ulceration or with a polypoid mass. However, metastatic gastric lesions which resemble early gastric carcinoma are rare. Typically, gastric metastases are confined to submucosa and muscularis, so that mucosal biopsy specimens might be false-negative. We report a case of gastric metastasis from infiltrative lobular carcinoma of the breast in a 66-year-old woman who had undergone left mastectomy with postoperative radiotherapy 17 years earlier. Initial diagnosis was early gastric carcinoma, signet ring cell type on gastric biopsy findings. However, definitive diagnosis of metastatic breast cancer was confirmed after endoscopic mucosal resection of a presumed primary early gastric carcinoma.

Angiosarcoma is a rare malignant tumor which occurs frequently in the skin and soft subcutis. Moreover, primary gastrointestinal angiosarcomas are very rare. This tumor manifests as non-specific symptoms such as gastrointestinal bleeding, abdominal pain and nausea. The diagnosis is often made at an advanced stage. Surgery, chemotherapy and radiotherapy are the mainstay of treatment. However, the prognosis is very poor. We report a case of primary angiosarcoma of the small intestine presenting as recurrent gastrointestinal bleeding. A 54-year-old man was admitted with recurrent gastrointestinal bleeding. An abdominal CT scan revealed an ileo-ileal intussusception. Segmental resection was performed with ileo-ileal anastomosis. The ileal mass was diagnosed as angiosarcoma on immunohistochemical stain. He received 3 cycles of chemotherapy, but died 5 months after the diagnosis.

Extramedullary plasmacytoma involves organs outside the bone marrow; however, involvement of the pancreas is rare. We recently experienced a case of extramedullary plasmacytoma of the pancreas that was diagnosed by endoscopic ultrasonography-guided fine needle aspiration (EUS-FNA). EUS-FNA, which has a high diagnostic accuracy and an excellent safety profile, is the modality of choice for establishing tissue diagnosis. We report a case of extramedullary plasmacytoma of the pancreas diagnosed using EUS-FNA.

A highly sensitive immunoassay was developed by using antibody-conjugated spherical mesoporous silica with immobilized enzymes. The higher ratio of enzyme/antibody than conventional ELISA improved both the sensitivity and dynamic range. Especially, the use of spherical mesoporous silica could achieve a limit of detection (LOD) with a sensitivity that is 20 times more than that of ELISA using amorphous silica.