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1.  Influence of Restricted Grazing Time Systems on Productive Performance and Fatty Acid Composition of Longissimus dorsi in Growing Lambs 
Fifty 3-month-old male Tan lambs (similar in body weight) were divided into 5 groups to investigate the effects of different restricted pasture grazing times and indoor supplementation on the productive performances and fatty acid composition of the intramuscular fat in growing lambs. The lambs grazed for different periods of time (12 h/d, 8 h/d, 4 h/d, 2 h/d, and 0 h) and received various amounts of supplementary feedings during the 120-day trial. Pasture dry matter intake (DMI), total DMI, average daily gains and the live body weights of the lambs were measured during the experiment. The animals were slaughtered at the end of the study, their carcass traits were measured, and their longissimus dorsi muscles were sampled to analyze the intramuscular fat (IMF) content and fatty acid profiles. The results indicated that the different durations of grazing and supplementary feedings affected the animal performances and the composition of fatty acids. Grazing for 8 h/d or 2 h/d with the corresponding supplementary concentrate resulted in lambs with higher body weights, carcass weights and IMF contents. Lambs with longer grazing times and less concentrate accumulated more healthy fatty acids such as conjugated linoleic acid and n-3 polyunsaturated fatty acid and had higher n-3/n-6 ratios. Overall, a grazing allowance of 8 h/d and the corresponding concentrate was recommended to maintain a high quantity and quality of lamb meat.
doi:10.5713/ajas.14.0937
PMCID: PMC4478478  PMID: 26104518
Grazing Time Allowance; Animal Performance; Fatty Acid Profile; Tan Lamb
2.  Correction: Estrogenicity of Glabridin in Ishikawa Cells 
PLoS ONE  2015;10(7):e0133340.
doi:10.1371/journal.pone.0133340
PMCID: PMC4505910  PMID: 26186551
3.  N-Glycan Branching Affects the Subcellular Distribution of and Inhibition of Matriptase by HAI-2/Placental Bikunin 
PLoS ONE  2015;10(7):e0132163.
The gene product of SPINT 2, that encodes a transmembrane, Kunitz-type serine protease inhibitor independently designated as HAI-2 or placenta bikunin (PB), is involved in regulation of sodium absorption in human gastrointestinal track. Here, we show that SPINT 2 is expressed as two species of different size (30-40- versus 25-kDa) due to different N-glycans on Asn-57. The N-glycan on 25-kDa HAI-2 appears to be of the oligomannose type and that on 30-40-kDa HAI-2 to be of complex type with extensive terminal N-acetylglucosamine branching. The two different types of N-glycan differentially mask two epitopes on HAI-2 polypeptide, recognized by two different HAI-2 mAbs. The 30-40-kDa form may be mature HAI-2, and is primarily localized in vesicles/granules. The 25-kDa form is likely immature HAI-2, that remains in the endoplasmic reticulum (ER) in the perinuclear regions of mammary epithelial cells. The two different N-glycans could, therefore, represent different maturation stages of N-glycosylation with the 25-kDa likely a precursor of the 30-40-kDa HAI-2, with the ratio of their levels roughly similar among a variety of cells. In breast cancer cells, a significant amount of the 30-40-kDa HAI-2 can translocate to and inhibit matriptase on the cell surface, followed by shedding of the matriptase-HAI-2 complex. The 25-kDa HAI-2 appears to have also exited the ER/Golgi, being localized at the cytoplasmic face of the plasma membrane of breast cancer cells. While the 25-kDa HAI-2 was also detected at the extracellular face of plasma membrane at very low levels it appears to have no role in matriptase inhibition probably due to its paucity on the cell surface. Our study reveals that N-glycan branching regulates HAI-2 through different subcellular distribution and subsequently access to different target proteases.
doi:10.1371/journal.pone.0132163
PMCID: PMC4501743  PMID: 26171609
4.  Lymph Node Metastasis, a Unique Independent Prognostic Factor in Early Gastric Cancer 
PLoS ONE  2015;10(7):e0129531.
Background
Lymph node metastasis (LNM) has been shown to be related to the prognosis of early gastric cancer (EGC). The choice of optimal treatment depends on an accurate pre-operative assessment of LNM status in EGC patients. However, in China, where EGC cases account for only a small part of gastric cancer (GC) cases, there are not enough data to make an accurate assessment. Therefore, this study, which involved a relatively large number of EGC patients, aimed to explore the relationship between clinicopathological characteristics and LNM in EGC.
Methods
Clinicopathological data from 205 EGC patients who underwent surgical resection at Sun Yat-Sen University Cancer Center from January 2000 to December 2011 were retrospectively analyzed. Clinicopathological characteristics were assessed to identify effective predictive factors for LNM and overall survival.
Results
LNM occurred in 52 (25.37%) EGC cases; of these cases, 18 occurred in intra-mucosal cancers (13 N1, 4 N2 and 1 N3), and 34 occurred in sub-mucosal cancers (22 N1, 7 N2 and 5 N3). Logistic regression analysis demonstrated that tumor differentiation (P=0.002), depth of tumor infiltration (P=0.004), vessel invasion (P=0.012), tumor size (P=0.020) and gender (P=0.022) were risk factors associated with LNM in EGC, listed in order of priority. The overall survival rate was 90.2%. Kaplan-Meier survival analysis showed that overall survival of EGC patients was significantly correlated with LNM (P=0.001), N staging (P<0.001) and invasion of lymphatic or blood vessels (P=0.010), but it was not correlated with tumor size, depth of tumor infiltration or tumor cell differentiation. Moreover, a multiple Cox regression analysis demonstrated that only N staging (P=0.001) could serve as an independent prognostic predictor in EGC patients.
Conclusions
Because LNM independently predicts the prognosis of EGC, endoscopic mucosal resection (EMR) or endoscopic submucosal dissection (ESD) and laparoscopic partial gastrectomy should be cautiously used in high-risk EGC patients. A pre-operative assessment of LNM status based on clinicopathological factors may be useful for therapy planning.
doi:10.1371/journal.pone.0129531
PMCID: PMC4496056  PMID: 26154617
5.  The Use of Unidirectional Barbed Suture for Urethrovesical Anastomosis during Robot-Assisted Radical Prostatectomy: A Systematic Review and Meta-Analysis of Efficacy and Safety 
PLoS ONE  2015;10(7):e0131167.
Background
Unidirectional barbed suture (UBS) has been widely used for surgery in recent years, especially for urethrovesical anastomosis (UVA) during robot-assisted radical prostatectomy (RARP). However, the efficacy and safety comparing it with conventional non-barbed suture (CS) for UVA is still controversial.
Aims
The objective of this study is to assess the current evidence regarding the efficacy and safety of UBS compared with CS for UVA during RARP.
Methods
We comprehensively searched PubMed, Embase, The Cochrane Library, SinoMed (Chinese) and other databases on Oct. 9, 2014 to conduct a systematic review and meta-analysis of all randomized controlled trials (RCTs) and other comparative studies evaluating these two types of suture. The outcome measures included anastomosis time operative time, posterior reconstruction (PR) time, postoperative leakage (PL) rate and continence rates at different time points (4-6 weeks, 3 months, 6-12 months) after surgery. Secondary outcomes included estimated blood loss (EBL) and length of catheterization (LOC).
Results
Three RCTs and six observational studies including 786 cases were identified. Meta-analysis of extractable data showed that use of UBS could significantly reduce anastomosis time (weighted mean difference [WMD]:-3.98min; 95% confidence interval [CI], -6.02 -1.95; p = 0.0001), operative time (WMD:-10.06min; 95% CI, -15.45–-4.67; p = 0.0003) and PR time (WMD:-0.93min; 95% CI, -1.52–-0.34; p = 0.002). No significant difference was found in PL rate, EBL, LOC, or continence rates at 4-6 weeks, 3 months and 6–12 months after surgery.
Conclusions
Our meta-analysis indicates that UBS appears to be safe and efficient as CS for UVA during RARP with not only shorter anastomosis time, operative time, PR time, but also equivalent PL rate, EBL, LOC, and continence rates at 4-6 weeks, 3 months and 6-12 months after surgery. For the inherent limitations of the eligible studies, future more persuasive RCTs are needed to confirm and update our findings.
doi:10.1371/journal.pone.0131167
PMCID: PMC4489906  PMID: 26135310
6.  High Fat Diet Produces Brain Insulin Resistance, Synaptodendritic Abnormalities and Altered Behavior in Mice 
Neurobiology of disease  2014;67:79-87.
Insulin resistance and other features of the metabolic syndrome are increasingly recognized for their effects on cognitive health. To ascertain mechanisms by which this occurs, we fed mice a very high fat diet (60% kcal by fat) for 17 days or a moderate high fat diet (HFD, 45% kcal by fat) for 8 weeks and examined changes in brain insulin signaling responses, hippocampal synaptodendritic protein expression, and spatial working memory. Compared to normal control diet mice, cerebral cortex tissues of HFD mice were insulin-resistant as evidenced by failed activation of Akt, S6 and GSK3β with ex-vivo insulin stimulation. Importantly, we found that expression of brain IPMK, which is necessary for mTOR/Akt signaling, remained decreased in HFD mice upon activation of AMPK. HFD mouse hippocampus exhibited increased expression of serine-phosphorylated insulin receptor substrate 1 (IRS1-pS616), a marker of insulin resistance, as well as decreased expression of PSD-95, a scaffolding protein enriched in post-synaptic densities, and synaptopodin, an actin-associated protein enriched in spine apparatuses. Spatial working memory was impaired as assessed by decreased spontaneous alternation in a T-maze. These findings indicate that HFD is associated with telencephalic insulin resistance and deleterious effects on synaptic integrity and cognitive behaviors.
doi:10.1016/j.nbd.2014.03.011
PMCID: PMC4083060  PMID: 24686304
7.  Seropositivity for Avian Influenza H6 Virus among Humans, China 
Emerging Infectious Diseases  2015;21(7):1267-1269.
doi:10.3201/eid2107.150135
PMCID: PMC4480397  PMID: 26079934
avian influenza; H6N2; viruses; occupationally exposed populations; human infection; agricultural workers' diseases; poultry; China; seropositivity
8.  Integrin αvβ6 sustains and promotes tumor invasive growth in colon cancer progression 
AIM: To detect the mechanism by which colon tumor escapes the growth constraints imposed on normal cells by cell crowding and dense pericellular matrices.
METHODS: An immunohistochemical study of integrin αvβ6 and matrix metalloproteinase-9 (MMP-9) was performed on tissue microarrays of 200 spots, including 100 cases of colon tumors.
RESULTS: High immunoreactivity for αvβ6 (73.7%; 28/38) and MMP-9 (76.5%; 52/68) was observed in invasive tumor portions. Furthermore, the effects of integrin αvβ6 on tumor invasive growth in nude mice were detected. Tumor invasive growth and high expression of both αvβ6 and MMP-9 were only seen in tumors resulting from WiDr cells expressing αvβ6 in the tumorigenicity assay. Flow cytometry was applied to analyze αvβ6 expression in colon cancer WiDr and SW480 cells. The effects of cell density on αvβ6 expression and MMP-9 secretion were also detected by Biotrak MMP-9 activity assay and gelatin zymography assay. High cell density evidently enhanced αvβ6 expression and promoted MMP-9 secretion compared with low density.
CONCLUSION: Integrin αvβ6 sustains and promotes tumor invasive growth in tumor progression via a self-perpetuating mechanism. Integrin ανβ6-mediated MMP-9 secretion facilitates pericellular matrix degradation at high cell density, which provides the basis of invasive growth.
doi:10.3748/wjg.v21.i24.7457
PMCID: PMC4481440  PMID: 26139991
Colonic neoplasms; Integrin αvβ6; Matrix metalloproteinase-9; Invasive growth
9.  A unification of models for meta-analysis of diagnostic accuracy studies without a gold standard 
Biometrics  2014;71(2):538-547.
Several statistical methods for meta-analysis of diagnostic accuracy studies have been discussed in the presence of a gold standard. However, in practice, the selected reference test may be imperfect due to measurement error, non-existence, invasive nature, or expensive cost of a gold standard. It has been suggested that treating an imperfect reference test as a gold standard can lead to substantial bias in the estimation of diagnostic test accuracy. Recently, two models have been proposed to account for imperfect reference test, namely, a multivariate generalized linear mixed model (MGLMM) and a hierarchical summary receiver operating characteristic (HSROC) model. Both models are very flexible in accounting for heterogeneity in accuracies of tests across studies as well as the dependence between tests. In this paper, we show that these two models, although with different formulations, are closely related and are equivalent in the absence of study-level covariates. Furthermore, we provide the exact relations between the parameters of these two models and assumptions under which two models can be reduced to equivalent submodels. On the other hand, we show that some submodels of the MGLMM do not have corresponding equivalent submodels of the HSROC model, and vice versa. With three real examples, we illustrate the cases when fitting the MGLMM and HSROC models leads to equivalent submodels and hence identical inference, and the cases when the inferences from two models are slightly different. Our results generalize the important relations between the bivariate generalized linear mixed model and HSROC model when the reference test is a gold standard.
doi:10.1111/biom.12264
PMCID: PMC4416105  PMID: 25358907
Diagnostic test; Generalized linear mixed model; Hierarchical model; Imperfect reference test; Meta-analysis
10.  Clinical impact of preoperative acute pancreatitis in patients who undergo pancreaticoduodenectomy for periampullary tumors 
AIM: To investigate the impact of preoperative acute pancreatitis (PAP) on the surgical management of periampullary tumors.
METHODS: Fifty-eight patients with periampullary tumors and PAP were retrospectively analyzed. Thirty-four patients who underwent pancreaticoduodenectomy (PD) and 4 patients who underwent total pancreatectomy were compared with a control group of 145 patients without PAP during the same period.
RESULTS: The preoperative waiting time was significantly shorter for the concomitant PAP patients who underwent a resection (22.4 d vs 54.6 d, P < 0.001) compared to those who did not. The presence of PAP significantly increased the rate of severe complications (Clavien grade 3 or higher) (17.6% vs 4.8%, P = 0.019) and lengthened the hospital stay (19.5 d vs 14.5 d, P = 0.006). A multivariate logistic regression analysis revealed that PAP was an independent risk factor for postoperative pancreatic fistula (OR = 2.91; 95%CI: 1.10-7.68; P = 0.032) and severe complications (OR = 4.70; 95%CI: 1.48-14.96; P = 0.009) after PD. There was no perioperative mortality.
CONCLUSION: PAP significantly increases the incidence of severe complications and lengthens the hospital stay following PD. PD could be safely performed in highly selective patients with PAP.
doi:10.3748/wjg.v21.i22.6937
PMCID: PMC4462735  PMID: 26078571
Pancreaticoduodenectomy; Complications; Preoperative pancreatitis; Pancreatic fistula
11.  Continuous Positive Airway Pressure Treatment Reduces Mortality in Elderly Patients with Moderate to Severe Obstructive Severe Sleep Apnea: A Cohort Study 
PLoS ONE  2015;10(6):e0127775.
Obstructive sleep apnea (OSA) is much more prevalent in older people than in middle-aged or young populations, and has been associated with cardiovascular disease. Continuous positive airway pressure (CPAP) is the first-line therapy for OSA, but its long-term clinical benefit in the elderly is unclear. Here, we carried out a prospective cohort study to explore the survival rate and incidence of cardiovascular events in elderly patients with moderate to severe OSA who did or did not receive CPAP treatment. The study included 130 patients (104 male, 26 female; mean age: 77.8 ± 6.2 years) who were followed up for a mean of 5 ± 2.54 years (range, 1–8 years). Thirty-six patients received CPAP and 88 had no CPAP. The results showed that mortality in the untreated group (21.6%) was significantly higher than in the CPAP group (5.6%). Kaplan–Meier survival analysis showed that the survival rate in the CPAP group was 94.4%, which was markedly higher than the rate of 78.4% in the untreated group. The incidence of cardiovascular events was 13.9% in the CPAP group and 55.7% in the untreated group. The present study provides evidence that CPAP can reduce mortality in older patients with moderate to severe OSA, and lead to a good long-term prognosis. The study also indicates that death in older OSA patients is associated with cardiovascular disease and diabetes.
doi:10.1371/journal.pone.0127775
PMCID: PMC4465781  PMID: 26068440
12.  The Structural Basis of Functional Improvement in Response to Human Umbilical Cord Blood Stem Cell Transplantation in Hearts with Post-Infarct LV Remodeling 
Cell transplantation  2013;24(6):971-983.
Cellular therapy for myocardial repair has been one of the most intensely investigated interventional strategies for acute myocardium infarction. Although the therapeutic potential of stem cells has been demonstrated in various studies, the underlying mechanisms for such improvement are poorly understood. In the present study, we investigated the long-term effects of stem cell therapy on both myocardial fiber organization and regional contractile function using a rat model of post-infarct remodeling. Human non-hematopoietic umbilical cord blood stem cells (nh-UCBSCs) were administered via tail vein to rats 2 days after infarct surgery. Animals were maintained without immunosuppressive therapy. In vivo and ex vivo MR imaging was performed on infarct hearts ten months after cell transplantation. Compared to the age-matched rats exposed to the identical surgery, both global and regional cardiac function of the nh-UCBSC-treated hearts, such as ejection fraction, ventricular strain and torsion, were significantly improved. More importantly, the treated hearts exhibited preserved fiber orientation and water diffusivities that were similar to those in sham-operated control hearts. These data provide the first evidence that nh-UCBSC treatment may prevent/delay untoward structural remodeling in post-infarct hearts, which supports the improved LV function observed in vivo in the absence of immunosuppression, suggesting a beneficial paracrine effect that occurred with the cellular therapy.
doi:10.3727/096368913X675746
PMCID: PMC4380875  PMID: 24332083
umbilical cord blood stem cells; ischemic heart injury; diffusion tensor MR imaging; displacement encoding; myocardial wall strain; fiber architecture
13.  Identification of a critical determinant that enables efficient fatty acid synthesis in oleaginous fungi 
Scientific Reports  2015;5:11247.
Microorganisms are valuable resources for lipid production. What makes one microbe but not the other able to efficiently synthesize and accumulate lipids is poorly understood. In the present study, global gene expression prior to and after the onset of lipogenesis was determined by transcriptomics using the oleaginous fungus Mortierella alpina as a model system. A core of 23 lipogenesis associated genes was identified and their expression patterns shared a high similarity among oleaginous microbes Chlamydomonas reinhardtii, Mucor circinelloides and Rhizopus oryzae but was dissimilar to the non-oleaginous Aspergillus nidulans. Unexpectedly, Glucose-6-phosphate dehydrogenase (G6PD) and 6-phosphogluconate dehydrogenase (PGD) in the pentose phosphate pathway (PPP) were found to be the NADPH producers responding to lipogenesis in the oleaginous microbes. Their role in lipogenesis was confirmed by a knockdown experiment. Our results demonstrate, for the first time, that the PPP plays a significant role during fungal lipogenesis. Up-regulation of NADPH production by the PPP, especially G6PD, may be one of the critical determinants that enables efficiently fatty acid synthesis in oleaginous microbes.
doi:10.1038/srep11247
PMCID: PMC4462047  PMID: 26059272
14.  Establishment and Validation of SSCLIP Scoring System to Estimate Survival in Hepatocellular Carcinoma Patients Who Received Curative Liver Resection 
PLoS ONE  2015;10(6):e0129000.
Background and Aims
There is no prognostic model that is reliable and practical for patients who have received curative liver resection (CLR) for hepatocellular carcinoma (HCC). This study aimed to establish and validate a Surgery-Specific Cancer of the Liver Italian Program (SSCLIP) scoring system for those patients.
Methods
668 eligible patients who underwent CLR for HCC from five separate tertiary hospitals were selected. The SSCLIP was constructed from a training cohort by adding independent predictors that were identified by Cox proportional hazards regression analyses to the original Cancer of the Liver Italian Program (CLIP). The prognostic performance of the SSCLIP at 12 and 36-months was compared with data from existing models. The patient survival distributions at different risk levels of the SSCLIP were also assessed.
Results
Four independent predictors were added to construct the SSCLIP, including age (HR = 1.075, 95%CI: 1.019–1.135, P = 0.009), albumin (HR = 0.804, 95%CI: 0.681–0.950, P = 0.011), prothrombin time activity (HR = 0.856, 95%CI: 0.751–0.975, P = 0.020) and microvascular invasion (HR = 19.852, 95%CI: 2.203–178.917, P = 0.008). In both training and validation cohorts, 12-month and 36-month prognostic performance of the SSCLIP were significantly better than those of the original CLIP, model of end-stage liver disease-based CLIP, Okuda and Child-Turcotte-Pugh score (all P < 0.05). The stratification of risk levels of the SSCLIP showed an enhanced ability to differentiate patients with different outcomes.
Conclusions
A novel SSCLIP to predict survival of HCC patients who received CLR based on objective parameters may provide a refined, useful prognosis algorithm.
doi:10.1371/journal.pone.0129000
PMCID: PMC4461285  PMID: 26057656
15.  DNA Marker Transmission and Linkage Analysis in Populations Derived from a Sugarcane (Saccharum spp.) x Erianthus arundinaceus Hybrid 
PLoS ONE  2015;10(6):e0128865.
Introgression of Erianthus arundinaceus has been the focus of several sugarcane breeding programs in the world, because the species has desirable traits such as high biomass production, vigour, ratooning ability and good resistance to environmental stresses and disease. In this study four genetic maps were constructed for two intergeneric populations. The first population (BC1) was generated from a cross between an Erianthus/Saccharum hybrid YC96-40 and a commercial sugarcane variety CP84-1198. The second population (BC2) was generated from a cross between YCE01-116, a progeny of the BC1 cross and NJ57-416, a commercial sugarcane cultivar. Markers across both populations were generated using 35 AFLP and 23 SSR primer pairs. A total of 756 and 728 polymorphic markers were scored in the BC1 and BC2 populations, respectively. In the BC1 population, a higher proportion of markers was derived from the Erianthus ancestor than those from the Saccharum ancestor Badila. In the BC2 population, both the number and proportion of markers derived from Erianthus were approximately half of those in the BC1 population. Linkage analysis led to the construction of 38, 57, 36 and 47 linkage groups (LGs) for YC96-40, CP84-1198, YCE01-116, and NJ57-416, encompassing 116, 174, 97 and 159 markers (including single dose, double dose and bi-parental markers), respectively. These LGs could be further placed into four, five, five and six homology groups (HGs), respectively, based on information from multi-allelic SSR markers and repulsion phase linkages detected between LGs. Analysis of repulsion phase linkage indicated that Erianthus behaved like a true autopolyploid.
doi:10.1371/journal.pone.0128865
PMCID: PMC4459986  PMID: 26053338
16.  Comparison of Biochemical Activities between High and Low Lipid-Producing Strains of Mucor circinelloides: An Explanation for the High Oleaginicity of Strain WJ11 
PLoS ONE  2015;10(6):e0128396.
The oleaginous fungus, Mucor circinelloides, is one of few fungi that produce high amounts of γ-linolenic acid (GLA); however, it usually only produces <25% lipid. Nevertheless, a new strain (WJ11) isolated in this laboratory can produce lipid up to 36% (w/w) cell dry weight (CDW). We have investigated the potential mechanism of high lipid accumulation in M. circinelloides WJ11 by comparative biochemical analysis with a low lipid-producing strain, M. circinelloides CBS 277.49, which accumulates less than 15% (w/w) lipid. M. circinelloides WJ11 produced more cell mass than that of strain CBS 277.49, although with slower glucose consumption. In the lipid accumulation phase, activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in strain WJ11 were greater than in CBS 277.49 by 46% and 17%, respectively, and therefore may provide more NADPH for fatty acid biosynthesis. The activities of NAD+:isocitrate dehydrogenase and NADP+:isocitrate dehydrogenase, however, were 43% and 54%, respectively, lower in WJ11 than in CBS 277.49 and may retard the tricarboxylic acid cycle and thereby provide more substrate for ATP:citrate lyase (ACL) to produce acetyl-CoA. Also, the activities of ACL and fatty acid synthase in the high lipid-producing strain, WJ11, were 25% and 56%, respectively, greater than in strain CBS 277.49. These enzymes may therefore cooperatively regulate the fatty acid biosynthesis in these two strains.
doi:10.1371/journal.pone.0128396
PMCID: PMC4457416  PMID: 26046932
17.  Aurora A orchestrates entosis by regulating a dynamic MCAK–TIP150 interaction 
Entosis, a cell-in-cell process, has been implicated in the formation of aneuploidy associated with an aberrant cell division control. Microtubule plus-end-tracking protein TIP150 facilitates the loading of MCAK onto the microtubule plus ends and orchestrates microtubule plus-end dynamics during cell division. Here we show that TIP150 cooperates with MCAK to govern entosis via a regulatory circuitry that involves Aurora A-mediated phosphorylation of MCAK. Our biochemical analyses show that MCAK forms an intra-molecular association, which is essential for TIP150 binding. Interestingly, Aurora A-mediated phosphorylation of MCAK modulates its intra-molecular association, which perturbs the MCAK–TIP150 interaction in vitro and inhibits entosis in vivo. To probe if MCAK–TIP150 interaction regulates microtubule plasticity to affect the mechanical properties of cells during entosis, we used an optical trap to measure the mechanical rigidity of live MCF7 cells. We find that the MCAK cooperates with TIP150 to promote microtubule dynamics and modulate the mechanical rigidity of the cells during entosis. Our results show that a dynamic interaction of MCAK–TIP150 orchestrated by Aurora A-mediated phosphorylation governs entosis via regulating microtubule plus-end dynamics and cell rigidity. These data reveal a previously unknown mechanism of Aurora A regulation in the control of microtubule plasticity during cell-in-cell processes.
doi:10.1093/jmcb/mju016
PMCID: PMC4034728  PMID: 24847103
Aurora A; TIP150; MCAK; entosis; microtubule plus-end; kinesin
19.  Risk scoring system and predictor for clinically relevant pancreatic fistula after pancreaticoduodenectomy 
AIM: To establish a scoring system to predict clinically relevant postoperative pancreatic fistula (CR-POPF) after pancreaticoduodenectomy (PD).
METHODS: The clinical records of 921 consecutive patients who underwent PD between 2008 and 2013 were reviewed retrospectively. Postoperative pancreatic fistula (POPF) was defined and classified by the international study group of pancreatic fistula (ISGPF). We used a logistic regression model to determine the independent risk factors of CR-POPF and developed a scoring system based on the regression coefficient of the logistic regression model. The optimal cut-off value to divide the risk strata was determined by the Youden index. The patients were divided into two groups (low risk and high risk). The independent sample t test was used to detect differences in the means of drain amylase on postoperative day (POD) 1, 2 and 3. The optimal cut-off level of the drain amylase to distinguish CR-POPF from non-clinical POPF in the two risk strata groups was determined using the receiver operating characteristic (ROC) curves.
RESULTS: Grade A POPF occurred in 106 (11.5%) patients, grade B occurred in 57 (6.2%) patients, and grade C occurred in 32 (3.5%) patients. A predictive scoring system for CR-POPF (0-6 points) was constructed using the following four factors: 1 point for each body mass index ≥ 28 [odds ratio (OR) = 3.86; 95% confidence interval (CI): 1.92-7.75, P = 0.00], soft gland texture (OR = 4.50; 95%CI, 2.53-7.98, P = 0.00), and the difference between the blood loss and transfusion in operation ≥ 800 mL (OR = 3.45; 95%CI, 1.92-7.75, P = 0.00); and from 0 points for a 5 mm or greater duct diameter to 3 points for a less than 2 mm duct (OR = 8.97; 95%CI: 3.70-21.77, P = 0.00). The ROC curve showed that the area under the curve of this score was 0.812. A score of 3 points was suggested to be the best cut-off value (Youden index = 0.485). In the low risk group, a drain amylase level ≥ 3600 U/L on POD3 could distinguish CR-POPF from non-clinical POPF (the sensitivity and specificity were 75% and 85%, respectively). In the high risk group, the best cut-off was a drain amylase level of 1600 (the sensitivity and specificity were 77 and 63%, respectively).
CONCLUSION: A 6-point scoring system accurately predicted the occurrence of CR-POPF. In addition, a drain amylase level on POD3 might be a predictor of this complication.
doi:10.3748/wjg.v21.i19.5926
PMCID: PMC4438027  PMID: 26019457
Pancreatic fistula; Pancreaticoduodenectomy; Postoperative complication; Risk factor; Logistic model
20.  Ligustrazine monomer against cerebral ischemia/reperfusion injury 
Neural Regeneration Research  2015;10(5):832-840.
Ligustrazine (2,3,5,6-tetramethylpyrazine) is a major active ingredient of the Szechwan lovage rhizome and is extensively used in treatment of ischemic cerebrovascular disease. The mechanism of action of ligustrazine use against ischemic cerebrovascular diseases remains unclear at present. This study summarizes its protective effect, the optimum time window of administration, and the most effective mode of administration for clinical treatment of cerebral ischemia/reperfusion injury. We examine the effects of ligustrazine on suppressing excitatory amino acid release, promoting migration, differentiation and proliferation of endogenous neural stem cells. We also looked at its effects on angiogenesis and how it inhibits thrombosis, the inflammatory response, and apoptosis after cerebral ischemia. We consider that ligustrazine gives noticeable protection from cerebral ischemia/reperfusion injury. The time window of ligustrazine administration is limited. The protective effect and time window of a series of derivative monomers of ligustrazine such as 2-[(1,1-dimethylethyl)oxidoimino]methyl]-3,5,6-trimethylpyrazine, CXC137 and CXC195 after cerebral ischemia were better than ligustrazine.
doi:10.4103/1673-5374.156991
PMCID: PMC4468780  PMID: 26109963
nerve regeneration; ligustrazine; ischemia; cerebral ischemia/reperfusion injury; neural regeneration
21.  Variability of Suitable Habitat of Western Winter-Spring Cohort for Neon Flying Squid in the Northwest Pacific under Anomalous Environments 
PLoS ONE  2015;10(4):e0122997.
We developed a habitat suitability index (HSI) model to evaluate the variability of suitable habitat for neon flying squid (Ommastrephes bartramii) under anomalous environments in the Northwest Pacific Ocean. Commercial fisheries data from the Chinese squid-jigging vessels on the traditional fishing ground bounded by 35°-45°N and 150°-175°E from July to November during 1998-2009 were used for analyses, as well as the environmental variables including sea surface temperature (SST), chlorophyll-a (Chl-a) concentration, sea surface height anomaly (SSHA) and sea surface salinity (SSS). Two empirical HSI models (arithmetic mean model, AMM; geometric mean model, GMM) were established according to the frequency distribution of fishing efforts. The AMM model was found to perform better than the GMM model. The AMM-based HSI model was further validated by the fishery and environmental data in 2010. The predicted HSI values in 1998 (high catch), 2008 (average catch) and 2009 (low catch) indicated that the squid habitat quality was strongly associated with the ENSO-induced variability in the oceanic conditions on the fishing ground. The La Niña events in 1998 tended to yield warm SST and favorable range of Chl-a concentration and SSHA, resulting in high-quality habitats for O. bartramii. While the fishing ground in the El Niño year of 2009 experienced anomalous cool waters and unfavorable range of Chl-a concentration and SSHA, leading to relatively low-quality squid habitats. Our findings suggest that the La Niña event in 1998 tended to result in more favorable habitats for O. bartramii in the Northwest Pacific with the gravity centers of fishing efforts falling within the defined suitable habitat and yielding high squid catch; whereas the El Niño event in 2009 yielded less favorable habitat areas with the fishing effort distribution mismatching the suitable habitat and a dramatic decline of the catch of O. bartramii. This study might provide some potentially valuable insights into exploring the relationship between the underlying squid habitat and the inter-annual environmental change.
doi:10.1371/journal.pone.0122997
PMCID: PMC4414546  PMID: 25923519
22.  Study on the behavioral changes of a post-stroke depression rat model 
The aim of this study was to observe the changes in the levels of learning and memory in a post-stroke depression (PSD) rat model. Forty-eight Wistar rats were randomly divided into three groups: Group A (normal group), group B (stroke group) and group C (PSD group). Each group comprised 16 rats. Three and 7 days after successful modeling, the open-field, sucrose solution consumption and forced swimming tests were performed, so as to evaluate behavioral changes in the three groups of rats. The sucrose solution consumption of the rats in group B was lower than that in group A in the tests at the two time-points, but only the difference after 7 days was statistically significant (P<0.05). The sucrose solution consumption of the rats in group C was lower than the consumption of those in groups A and B, and the differences were statistically significant (P<0.05). The open-field test results showed that the horizontal and vertical activity scores of the rats in group B were significantly lower than those of the rats in group A (P<0.05). Furthermore, the two test levels and the vertical activity scores of the rats in group C were significantly lower than those of the rats in groups A and B (P<0.05). The immobility time of the rats in the forced swimming test was higher in group B than that in group A at the two time-points, but the difference was only statistically significant after 7 days (P<0.05). The immobility time of the rats in group C was significantly higher than that in groups A and B (P<0.01). In conclusion, rats with PSD exhibited a significantly reduced memory capacity and altered behavior. The changes in the PSD rats were more severe than those in the rats in the stroke group.
doi:10.3892/etm.2015.2450
PMCID: PMC4487013  PMID: 26170928
stroke; post-stroke depression; rat model
23.  Treatment of renal cell carcinoma with a level III or level IV inferior vena cava thrombus using cardiopulmonary bypass and deep hypothermic circulatory arrest 
Background
The aim of this study was to investigate the minimally invasive cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA) approach in the management of renal cell carcinoma (RCC) with level III or IV inferior vena cava (IVC) thrombus and evaluate the survival outcomes.
Methods
We performed a retrospective analysis on 32 RCC patients with IVC thrombus that underwent nephrectomy and thrombectomy via the minimally invasive CPB/DHCA approach between January 2007 and December 2013. Perioperative variables (for example, operative time, CPB duration, and circulatory arrest duration), estimated blood loss, hospital stay, perioperative complications, and survival data were recorded and analyzed.
Results
Thirty-two patients (median age: 56 years) were treated surgically using the CPB and DHCA approach for RCC with a level III (n = 25) or level IV (n = 7) tumor thrombus. The median operation time was 360 min (interquartile range (IQR): 300 to 435 min) with median CPB and DHCA durations of 149 min and 23 min, respectively. The median estimated blood loss was 2,500 ml. Four complications were observed but no deaths occurred perioperatively. The median follow-up was 25 months (range: 4 to 64 months). The mean overall survival (OS) was 28.2 ± 4.6 months while the disease-free survival (DFS) was 19.5 ± 11.6 months. In patients with M0 disease, ten patients developed metastases and were treated with sorafenib as an adjuvant therapy. The mean OS and DFS of this subgroup were 25.4 ± 12.8 months and 16.0 ± 14.2 months, respectively.
Conclusions
Radical nephrectomy and thrombectomy using CPB and DHCA to treat RCC is a relatively safe approach associated with low morbidity and mortality. This minimally invasive procedure may help minimize surgical trauma and improve perioperative outcomes.
doi:10.1186/s12957-015-0584-8
PMCID: PMC4411871  PMID: 25897659
Cardiopulmonary bypass; Hypothermic arrest; Renal cell carcinoma; Thrombectomy; Tumor thrombus
24.  Is maximum primary tumor diameter still a prognostic factor in patients with nasopharyngeal carcinoma treated using intensity-modulated radiotherapy? 
BMC Cancer  2015;15:305.
Background
Intensity-modulated radiation therapy (IMRT) has represented a technical milestone that has facilitated the clinical implementation. The purpose of this study was to evaluate the prognostic value of maximum primary tumor diameter (MPTD) in patients with nasopharyngeal carcinoma (NPC) treated using IMRT.
Methods
Five-hundred and sixty-six patients with non-metastatic, histologically-confirmed NPC were retrospectively reviewed. MPTD was measured using magnetic resonance imaging (MRI). All patients were treated using IMRT; 87.5% (456/521) of patients with Stage T3-T4/N1-N3 disease also received cisplatin-based chemotherapy. Receiver operating characteristic (ROC) curves were used to identify the optimal MPTD cut-off point and examine the prognostic value of combining MPTD with the current T classification criteria.
Results
Median follow-up for all patients was 36 months (range, 1–52 months). The 3-year overall survival (OS), failure-free survival (FFS), distant metastasis-free survival (DMFS), and local relapse-free survival (LRFS) rates for patients with a MPTD ≤41 vs. >41 mm were 96.1% vs. 85.4%, 93.7% vs. 74.7%, 96.1% vs. 79.7%, and 98.1% vs. 92.9%, respectively (all P < 0.05). In multivariate analysis, MPTD was an independent prognostic factor for OS, FFS, DMFS and LRFS in all patients (all P < 0.05). Among stage T3-T4 patients, the 3-year OS, FFS, DMFS, and LRFS rates for patients with a MPTD ≤41 vs. >41 mm were 96.9% vs. 84.5%, 95.4% vs. 73.5%, 96.1% vs. 79.2%, and 99.3% vs. 92.6%, respectively (all P < 0.05). In multivariate analysis, MPTD was also an independent prognostic factor for OS, FFS and DMFS in stage T3-T4 patients (all P < 0.05), and the difference in LRFS was almost statistically significant (P = 0.05). ROC curves verified that inclusion of MPTD improved the predictive value of the current T classification criteria (P < 0.001).
Conclusions
MPTD was an independent prognostic factor in patients with NPC treated using IMRT, and significantly improved the prognostic value of the current T classification criteria for NPC.
doi:10.1186/s12885-015-1288-8
PMCID: PMC4406035  PMID: 25903652
Nasopharyngeal carcinoma; Maximum primary tumor diameter; Intensity-modulated radiotherapy; Prognostic factor; TNM stage
25.  Biliary tract intraductal papillary mucinous neoplasm: Report of 19 cases 
AIM: To gain a better understanding of biliary tract intraductal papillary mucinous neoplasm (BT-IPMN).
METHODS: From January 2000 to December 2013, 19 cases of BT-IPMN were retrospectively identified from a total of 343 biliary tract tumors resected in our single institution. Demographic characteristics, clinical data, pathology, surgical strategies, and long-term follow-up were analyzed.
RESULTS: The mean age of the 19 BT-IPMN cases was 53.8 years (range: 25-74 years). The most common symptom was abdominal pain (15/19; 78.9%), followed by jaundice (7/19; 36.8%). Cholangitis was associated with most (16/19; 84.2%) of the BT-IPMN cases. Macroscopically visible mucin was detected in all 19 patients, based on original surgical reports. The most common abnormal preoperative imaging findings for BT-IPMN were bile duct dilation (19/19; 100%) and intraluminal masses (10/19; 52.6%). Thirteen (68.4%) cases involved the intrahepatic bile duct and hilum. We performed left hepatectomy in 11/19 (57.9%), right hepatectomy in 2/19 (10.5%), bile duct resection in 4/19 (21.1%), and pancreatoduodenectomy in 1/19 (5.3%) patients. One (5.3%) patient was biopsied and received a choledochojejunostomy because of multiple tumors involving the right extrahepatic and left intrahepatic bile ducts. Histology showed malignancy in 10/19 (52.6%) patients. The overall median survival was 68 mo. The benign cases showed a non-significant trend towards improved survival compared to malignant cases (68 mo vs 48 mo, P = 0.347). The patient without tumor resection died of liver failure 22 mo after palliative surgery.
CONCLUSION: BT-IPMN is a rare biliary entity. Complete resection of the tumor is associated with good survival, even in patients with malignant disease.
doi:10.3748/wjg.v21.i14.4261
PMCID: PMC4394088  PMID: 25892877
Biliary tract; Cystic tumor; Intraductal papillary mucinous neoplasm; Mucinous tumor; Papillary tumor

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