A vascularized model of the human liver mimics regenerative responses.

Liver regeneration is a well-orchestrated process that is typically studied in animal models. Although previous animal studies have offered many insights into liver regeneration, human biology is less well understood. To this end, we developed a three-dimensional (3D) platform called structurally vascularized hepatic ensembles for analyzing regeneration (SHEAR) to model multiple aspects of human liver regeneration. SHEAR enables control over hemodynamic alterations to mimic those that occur during liver injury and regeneration and supports the administration of biochemical inputs such as cytokines and paracrine interactions with endothelial cells. We found that exposing the endothelium-lined channel to fluid flow led to increased secretion of regeneration-associated factors. Stimulation with relevant cytokines not only amplified the secretory response, but also induced cell-cycle entry of primary human hepatocytes (PHHs) embedded within the device. Further, we identified endothelial-derived mediators that are sufficient to initiate proliferation of PHHs in this context. Collectively, the data presented here underscore the importance of multicellular models that can recapitulate high-level tissue functions and demonstrate that the SHEAR device can be used to discover and validate conditions that promote human liver regeneration.

Transient Support from Fibroblasts is Sufficient to Drive Functional Vascularization in Engineered Tissues.

Formation of capillary blood vasculature is a critical requirement for native as well as engineered organs and can be induced in vitro by co-culturing endothelial cells with fibroblasts. However, whether these fibroblasts are required only in the initial morphogenesis of endothelial cells or needed throughout is unknown, and the ability to remove these stromal cells after assembly could be useful for clinical translation. In this study, we introduce a technique termed CAMEO (Controlled Apoptosis in Multicellular Tissues for Engineered Organogenesis), whereby fibroblasts are selectively ablated on demand, and utilize it to probe the dispensability of fibroblasts in vascular morphogenesis. The presence of fibroblasts is shown to be necessary only during the first few days of endothelial cell morphogenesis, after which they can be ablated without significantly affecting the structural and functional features of the developed vasculature. Furthermore, we demonstrate the use of CAMEO to vascularize a construct containing primary human hepatocytes that improved tissue function. In conclusion, this study suggests that transient, initial support from fibroblasts is sufficient to drive vascular morphogenesis in engineered tissues, and this strategy of engineering-via-elimination may provide a new general approach for achieving desired functions and cell compositions in engineered organs.

Controlled Apoptosis of Stromal Cells to Engineer Human Microlivers.

Engineered tissue models comprise a variety of multiplexed ensembles in which combinations of epithelial, stromal, and immune cells give rise to physiologic function. Engineering spatiotemporal control of cell-cell and cell-matrix interactions within these 3D multicellular tissues would represent a significant advance for tissue engineering. In this work, a new method, entitled CAMEO (Controlled Apoptosis in Multicellular tissues for Engineered Organogenesis) enables the non-invasive triggering of controlled apoptosis to eliminate genetically-engineered cells from a pre-established culture. Using this approach, the contribution of stromal cells to the phenotypic stability of primary human hepatocytes is examined. 3D hepatic microtissues, in which fibroblasts can enhance phenotypic stability and accelerate aggregation into spheroids, were found to rely only transiently on fibroblast interaction to support multiple axes of liver function, such as protein secretion and drug detoxification. Due to its modularity, CAMEO has the promise to be readily extendable to other applications that are tied to the complexity of 3D tissue biology, from understanding in vitro organoid models to building artificial tissue grafts.

[Value of contrast - enhanced ultrasonography and acoustic radiation force impulse elastography in identification of boundary range and viability of hepatic alveolar echinococcosis].

OBJECTIVE: To investigate the biologic viability and boundary range of hepatic alveolar echinococcosis (HAE) by the contrast-enhanced ultrasonography (CEUS) and acoustic radiation force impulse elastography (ARFI). METHODS: Totally 27 HAE patients confirmed by pathology underwent CEUS and ARFI examinations. RESULTS: Gray scale sonography of HAE showed unclear boundary, inhomogeneous, and middle hyperechoic nodules, and the maximum area was (6.08 ± 4.47) cm2 in 27 lesions. CEUS of HAE showed non-enhancement in three phases and black hole sign. Circumferential enhancement on the pe riphery of the lesion was synchronized with the liver parenchyma and showed "fast in and slow out". The maximum area was (8.87 ± 4.83) cm2. The area of ECUS was larger than gray scale sonography in HAE (t = 2.20, P = 0.03). The mean shear wave velocities (SWVs) of the interior, the boundary range, and the surrounding liver tissues of HAE were statistically different by ARFI (F = 84.538, P < 0.001), and the interior had the highest values. CONCLUSIONS: CEUS and ARFI examinations can detect the biologic viability and boundary range of migrating zone around HAE, which is valuable for guiding treatment, judging curative effect, and predicting prognosis.

Vascular Tissue Engineering: Progress, Challenges, and Clinical Promise.

Although the clinical demand for bioengineered blood vessels continues to rise, current options for vascular conduits remain limited. The synergistic combination of emerging advances in tissue fabrication and stem cell engineering promises new strategies for engineering autologous blood vessels that recapitulate not only the mechanical properties of native vessels but also their biological function. Here we explore recent bioengineering advances in creating functional blood macro and microvessels, particularly featuring stem cells as a seed source. We also highlight progress in integrating engineered vascular tissues with the host after implantation as well as the exciting pre-clinical and clinical applications of this technology.

[Factors associated with smoking cessation attempts in male current smokers in rural area of Shandong province].

Objective: To explore the influencing factors for the smoking cessation attempts in male current smokers. Methods: This cross-sectional study was conducted in nine villages selected from three counties (Pingyin, Junan and Liangshan) of Shandong province in August, 2010 through household questionnaire survey in villagers aged ≥15 years to collect the information about current smokers demographic characteristics and smoking-related behaviors. A hurdle count data model was used to assess factors associated with the times of past smoking cessation attempts. Results: Among 1 798 male current smokers, 29.53% had at least one smoking cessation attempt. Smokers who were married (ß=0.705, P=0.002), had high educational level (ß=0.214, P=0.026) and had higher level of awareness of smoking risks (ß=0.237, P=0.009) were more likely to have smoking cessation attempt. Young age at smoking initiation (ß=-0.167, P=0.035) and higher level of awareness of smoking risks (ß=0.146, P=0.020) were associated with increased smoking cessation attempts. Conclusions: Less male current smokers had smoking cessation attemps in rural area in Shandong. The factors influencing smoking cessation attempt varied. It is necessary to conduct targeted intervention according to the smokers' smoking cessation experience.

Biomimetic on-a-chip platforms for studying cancer metastasis.

Cancer metastasis is a multi-step, secondary tumor formation that is responsible for the vast majority of deaths in cancer patients. Animal models have served as one of the major tools for studying metastatic diseases. However, these metastasis models inherently lack the ability to decouple many of the key parameters that might contribute to cancer progression, and therefore ultimately limit detailed, mechanistic investigation of metastasis. Recently, organ-on-a-chip model systems have been developed for various tissue types with the potential to recapitulate major components of metastasis. Here, we discuss recent advances in in vitro biomimetic on-a-chip models for cancer metastasis.

A unique surface-initiated property of nanoparticles and application for the synthesis of hybrid organic-inorganic nanoparticles.

We report a unique property of nanoparticles to initiate acrylic acid and acrylamide solution polymerization under low air pressure conditions. This property could be applied to synthesize a wide variety of hybrid organic-inorganic nanoparticles, which hold great promise for use in nanophotonics, catalysis, and medical applications.

Comparison of RECIST 1.0 and RECIST 1.1 on computed tomography in patients with metastatic colorectal cancer.

OBJECTIVE: We conducted this study to compare tumor measurement by computed tomography (CT) and tumor response assessment between Response Evaluation Criteria In Solid Tumors (RECIST) 1.0 and RECIST 1.1 in patients with metastatic colorectal cancer (CRC). METHODS: We reviewed the medical records of patients with metastatic CRC who received first-line chemotherapy between January 2004 and December 2012 and compared CT tumor measurement using two RECIST versions. RESULTS: A total of 58 patients who had target lesions according to RECIST 1.0 were included in the study. The number of target lesions recorded by RECIST 1.1 was significantly lower than that by RECIST 1.0, with a decrease experienced in 48 patients (82.7%). Six patients had no target lesions because of the new criteria of RECIST 1.1 for lymph node size. Out of 95 lymph nodes from 58 patients, only 40% were defined as target lesions according to RECIST 1.1. The overall response rate of first-line chemotherapy according to RECIST 1.0 and 1.1 was 41.5 and 40.4%, respectively. The best tumor responses showed almost perfect agreement between RECIST 1.1 and RECIST 1.0 (ĸ = 0.913). Three patients showed disagreement of the best responses between the two RECIST versions. CONCLUSION: RECIST 1.1 showed a highly concordant response assessment with RECIST 1.0 in metastatic CRC and its clinical impact on therapeutic decisions was minimal.

Effects of in ovo injection of carbohydrate solution on small intestine development in domestic pigeons (Columba livia).

The objective of this study was to test the hypothesis that in ovo injection of carbohydrates into pigeon (Columba livia) amnion may improve the small intestine development. At d 14.5 of incubation, 80 fertile eggs were injected with 200 µL of carbohydrate solution, and 80 control eggs were not injected. The carbohydrate solution (wt/vol) contained 2.5% maltose + 2.5% sucrose, all dissolved in 0.75% saline. Twelve eggs from each treatment were randomly sampled at d 16 of incubation and the day of hatch, embryos or young pigeons were euthanized, and the jejunum samples were collected. Jejunal villus surface area, activity of the brush border enzymes, sucrase, maltase, aminopeptidase-N, and alkaline phosphatase, and mRNA expression of the digestion-absorption related genes oligopeptide transporter 1, sodium glucose transporter 1, glucose transporter 2, aminopeptidase-N, and sucrase-isomaltase were examined. Results showed that in ovo injection of carbohydrate solution caused a villus surface area increase of 38% on d 16 of incubation and 23% on day of hatch relative to controls (P < 0.05). The in ovo injected pigeons exhibited greater (P < 0.05) activities of jejunal sucrase, maltase, and alkaline phosphatase from d 16 of incubation to day of hatch compared with the controls. At day of hatch, aminopeptidase-N activity in embryos injected in ovo was approximately 27% greater (P < 0.05) than control embryos. Enhanced expressions of the jejunal sodium glucose transporter 1, glucose transporter 2, and aminopeptidase-N mRNA were found at d 16 of incubation in embryos that received carbohydrate solution into the amniotic fluid in comparison with the control group (P < 0.05). These results indicate that the in ovo injected pigeon may hatch with more mature enterocytes and greater intestinal digestive and absorptive capacity than the conventional hatchling. Therefore, the in ovo injected pigeons may become more precocial at hatch and easier to hand-rear during the immediate posthatch period.

Correlation between clinical diagnosis based on RDC/TMD and MRI findings of TMJ internal derangement.

This study was conducted to investigate the value of magnetic resonance imaging (MRI) in the diagnostic process based on the Research Diagnostic Criteria for Temporomandibular Disorder (RDC/TMD) by evaluating agreement between RDC/TMD and MRI diagnosis of disc displacement (DD) and correlation amongst MRI findings such as DD, joint effusion (JE), degenerative change and superior lateral pterygoid muscle (SLPM) attachment. Randomly selected MRIs of 200 joints from 100 TMD patients differentiated into RDC/TMD group II representing DD by clinical examination were reviewed retrospectively. The results show that Cohen's kappa value was 0.336 showing overall disagreement between RDC/TMD group II and MRI diagnoses (P<0.001). The Cohen's kappa value for group IIa, DD with reduction (DDWR), was -0.223 (P<0.01) showing disagreement, whilst the value was 0.546 for group IIb, DD without reduction (DDWOR) with limited opening, and 0.490 for group IIc, DDWOR without limited opening, showing moderate agreement (P<0.001). JE was detected with a higher probability as the state of DD advanced (P<0.001) and when degenerative joint changes were present (P<0.05). The difference of DD according to SLPM attachment was insignificant. MRI could be used when clinical examination cannot predict the true position of the disc.

Structural analysis of enniatin H, I, and MK1688 and beauvericin by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and their production by Fusarium oxysporum KFCC 11363P.

The molecular structures of enniatins H, I, and MK1688 and beauvericin were investigated by liquid chromatography-tandem mass spectrometry (LC-MS/MS). MS fragmentation occurred by loss of -CO after opening of the cyclic molecule to carbonyl carbon, and cleavage of the peptide and ester bonds in the molecular structure. Fusarium oxysporum KFCC 11363P was tested for its ability to produce beauvericin and enniatins H, I, and MK1688 on five cereal substrates: rice, barley, maize, wheat, and Indian millet kernels. Furthermore, optimal conditions for the production of the four mycotoxins by the Fusarium isolate were examined on maize at four temperatures (15, 20, 25, and 30 degrees C) and at three moisture contents (10, 20, and 40%). Large amounts of beauvericin and enniatin H were present in maize cultures at 25 degrees C (232.4 and 196.4 microg g(-1), respectively). Enniatins I and MK1688 were maximally formed at 20 degrees C (221.5 and 180.2 microg g(-1), respectively). The optimal moisture contents for the production of enniatins H (196.4 microg g(-1)) and MK1688 (165.6 microg g(-1)), were 40%.

Phase II study with oxaliplatin and S-1 for patients with metastatic colorectal cancer.

BACKGROUND: To evaluate the efficacy and safety of the combination of oxaliplatin and S-1 (OS) in treating metastatic colorectal cancer. PATIENTS AND METHODS: Eligible patients were those with measurable lesions, no previous history of chemotherapy (except adjuvant chemotherapy), an age of 18-70 years, and an Eastern Cooperative Oncology Group performance status of zero to two. Oxaliplatin 130 mg/m(2) was administered i.v. on day 1, and S-1 40 mg/m(2) b.i.d. was administered orally on days 1-14, every 3 weeks. RESULTS: Forty-eight patients (median age, 56 years) were enrolled: 23 had colon cancer, seven rectosigmoid colon cancer; and 18 rectal cancer. Of the 48 patients, 31 were diagnosed with metastatic cancer and 17 had relapsed cancer after surgery, with adjuvant chemotherapy or chemoradiotherapy. In total, 413 cycles were administered (median 6 per patient; range 2-24). Toxicity was evaluated in 48 patient and response in 46. Major toxic effects were grade 3/4 thrombocytopenia (13%) and neutropenia (10%). The overall response rate was 54% [95% confidence interval (CI) 40% to 68%]. The median time to progression and median survival time were 8.5 (95% CI 6.2-10.9) months and 27.2 (95% CI 20.3-34.0) months, respectively. CONCLUSIONS: These data indicate that the OS regimen is effective and well tolerated in patients with advanced colorectal cancer.

Clinical heterogeneity of extranodal NK/T-cell lymphoma, nasal type: a national survey of the Korean Cancer Study Group.

BACKGROUND: This national survey was undertaken to propose the classification of extranodal natural killer (NK)/T-cell lymphoma (NTCL) subtypes and to clarify a clinical heterogeneity. PATIENTS AND METHODS: Two hundred and eighty patients newly diagnosed as NTCL were enrolled from 22 Korean medical centers. Two subsets were compared: one involving the upper aerodigestive tract (UAT) and another involving the non-upper aerodigestive tract (NUAT) region, which comprises the skin, gastrointestinal tract, and liver or soft tissues. Clinical prognostic factors, survival outcomes, and independent predictors for survival were compared between each subset. RESULTS: NUAT-NTCL (59 patients) had significantly higher proportions of disseminated disease, aggressive biologic features, and unfavorable host reactions compared with UAT-NTCL (221 patients). NUAT-NTCL had shortened 5-year overall survival (OS) (22% versus 41%, P = 0.001). Ann Arbor staging, the International Prognostic Index, and the NTCL prognostic index failed to predict the OS of NUAT-NTCL, but did predict the OS in UAT-NTCL. Independent predictors for OS by multivariate analyses differed between each subset. In the NUAT subset, extranodal sites and regional nodes predicted the OS, while Ann Arbor staging, age, performance status, and lactate dehydrogenase level predicted the OS in the UAT subset. CONCLUSION: NUAT-NTCL may represent a distinctive disease entity in terms of clinical factors, independent predictors, and survival outcomes.

Regulation of adipocyte differentiation by PEGylated all-trans retinoic acid: reduced cytotoxicity and attenuated lipid accumulation.

Obesity is major risk factor for many disorders, including diabetes, hypertension and heart disease. Unfortunately, there is a dearth of therapeutic agents available to clinicians for the treatment of obesity. The principal aim of this study was to investigate whether PEGylated all-trans retinoic acid (PRA) can have favorable stability and biological activity in 3T3-L1 preadipocytes as an antiobesity drug. Here, we found that PRA inhibits the process of adipogenesis, including survival of adipocytes and differentiation to mature adipocytes. The results showed that RA nanoparticles (NPs) were prepared by PEGylation; below 200 nm, PRA-NPs were obtained. Moreover, PRA decreased glycerol-3-phosphate dehydrogenase activity in 3T3-L1 preadipocytes by acting with major adipocyte marker proteins such as PPARgamma2, C/EBPalpha and aP2 modulators. Apoptosis, in addition, increased as the level of RA increased from 10 to 20 microM, whereas PRA reduced apoptosis with increasing concentrations. Our data suggest that PRA-NP has potential as an antiobesity drug carrier due to its small particle size and PEGylated core-shell structure. In addition, our results suggest that PRA inhibits the process of adipogenesis and may be developed to treat obesity. Based on these results, PRA is suitable for adipocyte studies, and an enhanced effect of PRA with adipocyte differentiation offers a challenging approach for pharmaceutical applications.

Interaction between GNB3 C825T and ACE I/D polymorphisms in essential hypertension in Koreans.

Essential hypertension (EH) is considered a typical polygenic disease, so the evaluation of gene-gene interactions rather than the determination of single gene effects is crucial to understanding any genetic influences. The G-protein beta3-subunit (GNB3) 825T allele, associated with enhanced G-protein signalling, is a strong candidate for interactions with polymorphisms, such as insertion/deletion (I/D) polymorphism of angiotensin I-converting enzyme (ACE) gene. We investigated whether there is an association between GNB3 C825T and ACE I/D polymorphisms for the development of EH. We carried out a case-control study of 688 hypertensive and 924 normotensive subjects recruited from South Korea. The GNB3 C825T and ACE I/D genotypes were determined by polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism methods, respectively. The distributions of alleles and genotypes for the GNB3 C825T and ACE I/D polymorphisms were not found to be significantly associated with hypertensive status in either males or females. Logistic regression analysis indicated that the GNB3 825T allele carriers were positively associated with EH in males (odds ratio (OR) for TT/CT, 1.459; 95% confidence interval (CI), 1.048-2.033, P=0.0255). In analysis of gene-gene interaction, we found that there was a significant interaction between the GNB3 825T and ACE D alleles (P<0.05). OR for EH was significantly higher in 825T allele carriers with ACE D allele (OR, 1.490; 95% CI, 1.117-1.987, P=0.0067). A significant interaction between the GNB3 825T and the ACE D alleles may contribute to the predisposing effect for the development of EH in Koreans.

Salvaging and maintaining non-maturing Brescia-Cimino haemodialysis fistulae by percutaneous intervention.

AIM: To report our experience of the salvage of non-maturing Brescia-Cimino dialysis fistulae by percutaneous intervention. MATERIALS AND METHODS: Twenty-two patients (15 men, 68%; mean age: 58 years range: 42-79) with non-maturing Brescia-Cimino fistulae were treated by percutaneous angioplasty. Fistulae were created a mean of 2.7 months (range, 1-13 months) before intervention. The size of balloons used was 4mm for the arterial and anastomotic stenosis and 5mm or 6mm for the venous stenosis. RESULTS: On initial venography, venous stenosis (17 fistulae) or occlusions (five fistulae) were responsible for non-maturation. Stenoses or occlusions were adjacent to the arterial anastomoses in 18 patients and in the venous outflow (future puncture zone) in four patients. Additionally, a focal arterial stenosis was present in one and occlusion of the innominate vein in one other patient. Clinical success (initiation of dialysis) was achieved in 21/22 patients (95.5%). Twelve patients required 18 repeat angioplasties for recurrent stenosis. Two patients had small extravasation that required no further treatment. Over a follow-up period of 5-40 months (mean 14.6 months) 12 patients required repeat angioplasty. The mean interval between the initial angioplasty and subsequent intervention was 7.5 month (range 3-12 months). Primary patency after intervention at 6 and 12 months was 82 and 28%. Secondary patency at 6 and 12 months was 95 and 85%. CONCLUSION: Percutaneous intervention can effectively salvage non-maturing Brescia-Cimino fistulae. As repeat angioplasty is often necessary to maintain function, careful surveillance is necessary.

Release of ciprofloxacin from chondroitin 6-sulfate-graft-poloxamer hydrogel in vitro for ophthalmic drug delivery.

The system was designed to use Poloxamer as a vehicle for ophthalmic drug delivery using in situ gel formation property. To enhance the wound healing and cell adhesion as well as transparency of Poloxamer hydrogel, chondroitin 6-sulfate (C6S) was introduced into Poloxamer. For this purpose, mono amine-terminated Poloxamer (MATP), which was end-capped with ethylene amine group only in one side of terminal hydroxyl groups of Poloxamer, was synthesized. Subsequently, C6S-graft-Poloxamer copolymer (C6S-g-Poloxamer) was prepared by reaction between the amine groups of MATP and carboxyl groups of C6S in the presence of 1-ethyl-3-(3-dimethylaminopropyl)-carboimide (EDC). The coupling of MATP with C6S was clarified by 1H-NMR and FT-IR spectroscopy. The gelation temperature of graft copolymers was determined by measuring the temperature at which immobility of the meniscus in each solution was first noted. Release behavior of ciprofloxacin from C6S-g-Poloxamer hydrogel in vitro was investigated as a function of C6S content in the graft copolymer by a spectrophotometric assay at 287 nm using an UV spectrophotometer. Differences in the adhesion and morphology of human lens cell between Poloxamer- and C6S-g-Poloxamer-coated surfaces were also investigated. The gelation temperatures of C6S-g-Poloxamer copolymers were lowered with increasing of the concentration of the copolymer and decreasing of C6S content. The release of ciprofloxacin from the graft copolymer was sustained compared with Poloxamer itself and decreased with increasing the content of C6S in the copolymer due to the in situ gel formation of the copolymer and viscous properties of C6S. Human lens cells (B3) adhered to C6S-g-Poloxamer-coated surface were observed as transformed shapes after 2 days. The bioadhesive and thermally gelling of these graft copolymers will be expected to be an excellent drug carrier for the prolonged delivery to surface of the eye.

Identification of a tobacco protein interacting with tomato mosaic virus coat protein and facilitating long-distance movement of virus.

The protein-protein interaction of virus and host is essential for virus infection and host defense. The coat protein (CP) of tomato mosaic virus (ToMV) has been proved to be involved in cell-to-cell and long-distance movements of viruses that are presumably related with the protein-protein interactions. However, the host proteins that interact with the ToMV coat protein (ToCP) are largely unknown. In this study, we isolated a cDNA from a tobacco library through yeast two-hybrid system, which encodes a protein designated the ToMV CP-interacting protein-L (IP-L) that interacted with ToCP in vitro and in vivo. Sequencing analysis revealed that the putative coding region of IP-L gene was identical to that of an 'elicitor responsive protein' gene from N. tabacum (Genbank: #AB040409). A homology was also found between the cDNA sequence of IP-L and two senescence-related cDNAs (SENU1: Z75523 and AY479987) isolated from tomato and pepper. Northern blotting analysis showed that the mRNA level of IP-L was elevated after infection of ToMV. Then, we investigated the in vivo function of IP-L using virus-induced gene silencing (VIGS) and virus challenging assay. Semi-quantitative RT-PCR and Northern blotting results showed that the endogenous mRNA of IP-L in N. benthamiana plant was silenced at 10 days post inoculation with the in vitro transcripts of PVX-IP-L that were produced from the potato virus X (PVX)-based gene silencing plasmid pPC2S.IP-L. The IP-L silent plant developed a delayed systemic symptom at 7 days post challenging with ToMV, indicating that a high expression of IP-L was necessary for the interaction with ToCP to assist the viral transportation. Together, our data suggested that IP-L is a novel plant factor that interacts with the coat protein of ToMV and facilitates the long-distance movement of virus, which may provide a valuable clue for us to further investigate the mechanisms of plant virus infection and to control plant virus diseases.