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BACKGROUND: Neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and monocyte-to-lymphocyte ratio (MLR) are all markers of systemic inflammation response. The role of systemic inflammation in the development of esophageal fistula (EF) has yet to be defined. This study aimed to investigate the predictive value of hematologic measures of inflammation and to set up a predictive model. METHODS: The data of esophageal cancer (EC) patients who received chemoradiotherapy (CRT) in our institution between January, 2015 and January, 2018 were retrospectively collected. The NLR, PLR, and MLR of these enrolled patients were calculated. Univariate and multivariate analyses were performed to find the independent risk factors of EF. Moreover, a nomogram model was developed to predict the probability of fistula occurring in EC patients. RESULTS: For PLR, the optimal cut-off value was 153. Patients with PLR >153 had a higher probability of developing fistula than those with PLR ≤153 (P

BACKGROUND The poor palatability, low digestibility, and unpleasant color of parboiled rice (PR) have severely hampered its acceptance by consumers. It is hence necessary and urgent to develop a new method for producing high-quality PR. In the current study, the effect of high hydrostatic pressure (HHP) pre-soaking on the color, textural properties, and the degree of retrogradation of PR was investigated. RESULTS With HHP from 100 to 500 MPa, the water adsorption rate increased and cooking time decreased. Parboiled rice samples presented higher lightness scores (L) and had lower color intensity (B). Compared with a control group, PR samples treated with high-pressure pre-soaking showed a reduction of hardness values from 0.69% to 32.99%, and gumminess values also decreased from 8.58% to 33.62%. The differential scanning calorimetry (DSC) results indicated that the enthalpy values of PR samples decreased after high pressure pre-soaking. The molecular structure of PR characterized by Fourier transform infrared spectrometry confirmed that HHP pre-soaking could decrease the retrogradation level. CONCLUSION The findings outlined above suggest that the texture and retrogradation properties of PR were improved after high-pressure pre-soaking. © 2021 Society of Chemical Industry.

Gene therapy is a promising strategy for treating ischemic disease by solving the dual dilemma of ischemia and inflammation. However, its development remains limited by inefficient gene transfection. Hence, we propose a "dual genes + all-adaptive carrier" idea. We have innovatively co-delivered eNOS gene and the ZNF

Aim: To evaluate long-term outcome and prognostic factors of stage III esophageal cancer after definitive radiotherapy using three dimensional conformal radiotherapy (3DCRT) or intensity-modulated radiotherapy (IMRT) techniques. Methods: Patients with T3N1M0/T4N0-1M0 esophageal squamous cell carcinoma (ESCC) treated with definitive radiotherapy from 2002 to 2016 in 10 Chinese medical centers were retrospectively analyzed. Overall survival (OS) and progression-free survival (PFS) rates were calculated. Prognostic factors were analyzed by Log-rank test and multivariable Cox model. Results: Survival data of 1,450 patients were retrospectively collected. With a median follow-up time of 65.9 months, 1-, 3-, and 5-year OS rates were 69.3, 36.7, and 27.7%, respectively, and PFS rates were 58.6, 32.7, and 27.4%, respectively. Univariable analyses showed that gender, age, lesion location, lesion length, largest tumor diameter, lymph node metastasis, gross tumor volume, EQD2, short-term response, and concurrent chemotherapy were prognostic factors for OS. Multivariable analyses showed that lesion location, T-classification, GTV size, EQD2, and short-term response to RT were independent prognostic factors for OS, and tumor diameter, GTV size, and short-term response were independent prognostic factors for PFS. Conclusions: This study demonstrated that definitive radiotherapy using 3DCRT and IMRT provides promising outcomes for locally advanced ESCC.

Esophageal cancer (EC) has the seventh highest incidence and the sixth highest mortality rate of any type of cancer worldwide. In China, esophageal squamous cell carcinoma (ESCC) accounts for more than 95% of EC patients. The main treatment for EC patients is surgery and/or chemoradiotherapy (CRT). A large proportion of EC patients are already at an advanced stage of the disease by the time they are diagnosed. In these cases, CRT is left as the only treatment choice, and the treatment outcome is poor. Immune checkpoint inhibitors (ICIs) can improve clinical response and patient survival of patient with many types of tumors through reactivating antitumor immune response. The study of ICIs in ESCC is relative delayed compared with that in other solid tumors. Recent results from clinical trials have demonstrated the safety and efficacy of ICIs either alone or combined with chemotherapy or chemoradiotherapy in ESCC patients. Accumulated evidences also have shown the improved treatment outcome was associated with PD-L1 expression, tumor DNA instability-induced tumor mutational burden (TMB), and drawing lymphocytes into the tumor. Based on these findings, ICIs combined with CRT or radiotherapy (RT) are the focus of ongoing studies. This review will summarize the recent progress in this field, especially the mechanism of ICIs used in ESCC, their clinical efficacy and toxicities, and potential biomarkers.

BACKGROUND: We aimed to analyze the value of metal clip markers guided and placed by endoscopic ultrasonography (EUS) in the delineation of gross tumor volume (GTV) for thoracic esophageal squamous cell carcinoma. METHODS: From September 2016 to September 2018, patients with thoracic esophageal squamous cell carcinoma in Tianjin Medical University Cancer Institute and Hospital were recruited in the prospective trial, NCT02959385. They underwent titanium clips placement on tumor superior and inferior boundaries under EUS by a single expert endosonographer before radiotherapy computed tomography (CT) simulation. According to the clip markers, the reference GTVs were contoured by one experienced radiation oncologist. With the help of the Eclipse treatment planning system, clip markers on CT were concealed. Afterward, two other radiation oncologists with expertise in esophageal cancer delineated GTVs, defined as conventional GTVs, based on endoscopy and barium radiography findings. The two GTVs were compared and analyzed. Subgroup analysis was conducted in different T stage [early (T1 + T2) vs. advanced (T3 + T4)], focus location (upper vs. middle vs. lower segment), and tumor length (5 cm) groups. RESULTS: The trial recruited 55 patients with 60 thoracic esophageal cancer foci. A total of 111 titanium clips were guided and implanted by EUS. Before CT simulation, two titanium clips at two foci fell off. After the procedure, no case of intolerable esophageal pain, hemorrhage, or perforation occurred. Compared to reference GTVs’, discrepancies of conventional GTVs’ superior borders were 0.91±0.82 cm (P

The development of smart cities construction can solve the problems in the process of planning, construction and operation of cities, and it has gradually become a hot spot in China and the rest of the world. The concept and application of City Information Model (CIM) has developed rapidly since the concept was put forward, and it becomes an important part of the construction of smart city. It is regarded as the carrier of synchronic growth of physical city and digital twin city. However, the development of new things is not smooth sailing. Based on the author’s experience in the process of creating city information model, this paper summarizes the main problems in the construction of City Information Model platform and gives some suggestions for promoting the development of City Information Model, which can be referenced effectively for the research and practice of the construction of city information model.

Introduction: Intensive treatments can often not be administered to elderly patients with esophageal squamous cell carcinoma (ESCC), leading to a poorer prognosis. This multi-center phase II trial aimed to determine the toxicity profile and efficiency of S-1-based simultaneous integrated boost radiotherapy (SIB-RT) followed by consolidation chemotherapy with S-1 in elderly ESCC patients and to evaluate the usefulness of comprehensive geriatric assessment (CGA). Patients and Methods: We prospectively enrolled 46 elderly patients (age ≥ 70 years) with histopathologically proven ESCC. The patients underwent pretreatment CGA followed by SIB-RT (dose, 59.92 Gy/50.4 Gy) in 28 daily fractions administered using intensity-modulated radiotherapy or volumetric-modulated arc therapy. S-1 was orally administered (40-60 mg/m2) concurrently with radiotherapy and 4-8 weeks later, for up to four 3-week cycles at the same dose. Results: The median survival time was 22.6 months. The 1- and 2-year overall survival rates were 80.4 and 47.8%, respectively. The overall response rate was 78.3% (36/46). The incidence of grade 3-4 toxicities was 28% (13/46). The most common grade 3-4 toxicities were radiation esophagitis (5/46, 10.9%), nausea (4/46, 8.7%), anorexia (3/46, 6.5%), and radiation pneumonitis (3/46, 6.5%). There were no grade 5 toxicities. CGA identified that 48.8% of patients were at risk for depression and 65.5% had malnutrition. Conclusion: Concurrent S-1 treatment with SIB-RT followed by 4 cycles of S-1 monotherapy yielded satisfactory tumor response rates and manageable toxicities in selected elderly patients with ESCC. Pretreatment CGA uncovered numerous health problems and allowed the provision of appropriate supportive care. Clinical Trial Registration: www.ClinicalTrials.gov, identifier: NCT02979691.

Purpose: To assess the benefit of post-mastectomy radiotherapy (PMRT) in breast cancer (BC) patients with T1-2N1M0 who developed pathologically negative lymph nodes (ypN0) after undergoing neoadjuvant chemotherapy (NAC) and mastectomy. Patients and Materials: Patients with T1-2 tumors and positive lymph node(s) who became pN0 after NAC and mastectomy were screened from our prospectively maintained database. The primary endpoint was recurrence-free survival (RFS), and the secondary endpoints were local recurrence-free survival (LRFS) and overall survival (OS). Propensity-score matching (PSM) was conducted for the comparison between PMRT and non-PMRT groups. Results: Of the 142 eligible patients, 110 (77.5%) received PMRT, and 32 (22.5%) did not. The median follow-up time was 72 months. Univariate analyses showed that the 5-year RFS, LRFS, and OS rates were 88.7, 94.5, and 96.1, respectively, with PMRT and 72.4, 90.1, and 95.0% without PMRT (p = 0.028; p = 0.151; p = 0.971). Multivariate analyses established PMRT as a significant prognostic factor for RFS rate (HR, 0.411; 95% CI, 0.175–0.968; p = 0.042). After a PSM analysis (64 in the PMRT group vs. 32 in the non-PMRT group), PMRT remained significant, with improved RFS in univariate and multivariate analysis (with 5-year RFS rates of 90.1 vs. 72.4%, respectively, p = 0.016; HR, 0.323, 95%CI, 0.115–0.913, p = 0.033). In the subgroup of 48 (33.8%) patients with pathologic complete responses (pCR, ypT0, and ypN0) after NAC, PMRT did not affect RFS (HR, 0.226; 95% CI, 0.034–1.500; p = 0.123). Conclusions: PMRT might benefit pT1-2N1M0 patients with pN0 after NAC. Patients with pCR might consider omitting PMRT. Prospective studies are needed to assess the effect of PMRT on this specific patient population.

Ischemia-reperfusion (I/R) plays an important role in myocardial damage, which has been widely recognized as a key procedure in the cardiovascular disease. A hypoxia/reoxygenation (H/R) model was established using H9c2 cardiomyocytes to investigate the possible positive effect of oxymatrine (OMT), an alkaloid originating from the traditional Chinese herb Sophora flavescens Aiton, on cardiomyocytes exposed to H/R injury and the underlying molecular mechanisms. Cell viability was measured using the MTT assay, lactate dehydrogenase release measurements and hematoxylin and eosin staining. Oxidative stress was detected by measuring cellular malondialdehyde (MDA) content, as well as superoxide dismutase (SOD) and catalase (CAT) activities. Apoptosis was detected using TUNEL staining and flow cytometric analysis, and the underlying mechanism was investigated using reverse transcription-quantitative PCR and western blot analyses. The results revealed that OMT increased the viability of H9c2 cardiomyocytes exposed to H/R. The OMT pretreatment decreased the production of MDA by reactive oxygen species and increased the activities of SOD and CAT. Furthermore, the OMT pretreatment reduced the expression of Bax and caspase-3, while inducing Bcl-2 expression. In addition, the protective effect of OMT was shown to be associated with the PI3K/Akt signaling pathway, and the PI3K inhibitor LY294002 attenuated the effects of OMT on the H9c2 cardiomyocytes exposed to H/R. These findings indicate that OMT could be a potential therapeutic candidate for the treatment of myocardial ischemia/reperfusion injury.

Biocompatibility, stability and high efficiency profiles are critical points for promoting the practical applications of gene delivery systems. The incorporation of cell-penetrating peptides (CPPs), REDV, and a nuclear localization signal (NLS) peptide sequence has been considered to be a promising strategy for developing efficient gene carriers to transfect vascular endothelial cells (ECs). However, these integrated multifunctional peptide carriers are usually limited by their inefficient targeting function and weak endosomal escape ability. Aiming to develop more efficient gene carriers, the integrated multifunctional REDV-G-TAT-G-NLS-C sequence was conjugated to polyhedral oligomeric silsesquioxane (POSS) by heterobifunctional poly(ethylene glycol) in the current study. This star-shaped polymer carrier complexed with the pZNF580 plasmid to form gene complexes, and then the histidine-rich peptide of REDV-TAT-NLS-H12 (TP-H12) was incorporated into their surface to obtain ternary gene delivery systems with enhanced endosomal escape ability. These ternary gene delivery systems exhibited low cytotoxicity towards ECs and possessed high REDV-mediated cellular uptake, excellent internalization efficiency, rapid endosomal escape and high nucleus translocation capacity. The endosomal escape of the ternary complexes was improved due to the pH buffering capacity of the histidine residue in TP-H12 and the optimized macropinocytosis internalization pathway. Moreover, these CPP-based ternary gene delivery systems have high gene delivery efficiency and could improve the migration of ECs as demonstrated by gene expression and transwell assay. These systems may serve as a promising candidate for gene delivery and transfection in ECs, which is advantageous for EC migration and endothelialization on the biomaterial surface.

Purpose To evaluate the safety and efficacy of Cyberknife® (CK) for the treatment of primary or recurring thymic tumours. Materials and methods We retrospectively reviewed 12 patients (16 tumour lesions) with primary or recurring thymic tumours who were treated with CK between March 2008 and October 2017. Their data was stored in prospectively collected database. Kaplan–Meier method was used to calculate survival curves. Results Five patients (41.7%), who had inoperable disease or refused surgery, were treated with CK initially, and 7 patients (58.3%) were treated with CK when they had recurrence diseases. The disease sites treated with CK were primary tumour site (5), regional lymph nodes (4), tumour bed (3), chest wall (2), pleura (1), and bone (1). The median target volume was 43.8 cm3 (range, 13.1–302.5 cm3) for the 16 tumour lesions. The median follow-up time was 69.3 months (range, 9.7–124.8 months). The median survival time was 48.2 months, and the 5-year and 10-year OS rates were 68.2% and 45.5%, respectively. A high response rate for the tumour lesions irradiated with CK was obtained. Only one patient (8%) experienced in-field recurrence, and the 5-year local recurrence free survival was 90.9%. A case indicated that CK may induce the abscopal effect, which provides the potential to combine CK and immunotherapy. No severe radiation related toxicities were observed, and no treatment related death occurred. Conclusion CK treatment resulted in good outcomes, particularly local control, with minimal side effects, in highly selected patients with primary and recurring thymic tumours. More studies with larger sample are needed.

Redox-responsive cationic polymers have gained considerable attention in gene delivery due to low cytotoxicity and spatio-temporal release of DNA into the cells. Here, we reported the synthesis of reducible disulfide conjugated polyethyleneimine (1.8 kDa) (denoted as SS-PEI) and its application to transfer pEGFP-ZNF580 plasmid (pZNF580) into EA.hy926 cell. This reducible SS-PEI polymer was prepared by one-step polycondensation reaction of low molecular weight PEI with bis-(p-nitrophenyl)-3,3′-dithiodipropionate. The SS-PEI successfully condensed pZNF580 into nano-sized complexes (170 ± 1.5 nm to 255 ± 1.6 nm) with zeta potentials of 3 ± 0.4 mV to 17 ± 0.9 mV. The complexes could be triggered to release pZNF580 when exposed to the reducing environment of 5 mM dithiothreitol. Besides, the SS-PEI exhibited low cytotoxicity. In vitro transfection results showed that SS-PEI exhibited good transfection efficiency comparable to PEI25kDa. Thus, the SS-PEI could act as an reducible gene carrier with good transfection efficiency and low cytotoxicity.

Background Protecting the intestinal mucosa from being destroyed helps reduce the inflammation caused by acute pancreatitis (AP). In this study, whether okra pectin (OP) could attenuate the inflammation of AP through protecting the intestinal barrier was investigated. Results OP was obtained from crude okra pectin (COP) through the purification by DEAE cellulose 52 column. Supplementation with OP or COP in advance reduced the severity of AP, as revealed by lower serum amylase and lipase levels, abated pancreatic edema, attenuated myeloperoxidase activity and pancreas histology. OP or COP inhibited the production of pancreatic proinflammatory cytokines, including tumor necrosis factor-α and interleukin-6. In addition, the upregulation of AP-related proteins including ZO-1, occludin, the antibacterial peptide-defensin-1 (DEFB1) and cathelicidin-related antimicrobial peptide (CRAMP), as well as the histological examination of colon injuries, demonstrated that OP or COP provision could effectively maintain intestinal barrier function. Ultimately, dietary OP or COP supplementation could inhibit AP-induced intestinal inflammation. For the above, the effect of OP was better than COP. Conclusion Dietary OP supplementation could be considered as a preventive method that effectively interferes with intestinal damage and attenuates inflammatory responses trigged by AP. © 2020 Society of Chemical Industry.

In this study, the preparation of molecularly imprinted polymers for bilobalide (BBMIPs) was successfully achieved by bulk polymerization with methacrylamide (MAM), trimethylolpropane triacrylate (TMPTA), and acetonitrile (ACN) as functional monomer, cross-linker, and solvent, respectively. After Gaussian software simulation and single factor experiments, the prepared MIPs with a molar ratio of 1:4:15 for BB-MAM-TMPTA were systematically characterized. The hydrogen bonding interaction between BB and MAM was confirmed by a combination of FTIR and NMR analysis. Thermal gravimetric analysis results displayed that MIPs have excellent thermal stability under high temperature. Additionally, the average pore size and surface area of MIPs were found to be higher than those of NIPs through nitrogen adsorption results. The results of static adsorption and kinetic adsorption suggested that the adsorption equilibrium concentration was 0.6 mg/mL and the equilibrium time was 5 h, and the Langmuir and pseudo-second-order kinetic models were proven to fit with static and kinetic adsorption behaviors, respectively. Meanwhile, the selective adsorption study revealed that MIPs show high adsorption and great selectivity towards BB in comparison with other substances having similarly structure. MIPs also possessed a good performance on reusability, maintaining a high recovery rate after being reused 5 times. The application experiment further indicated that MIPs can effectively separate BB from low purity samples. Therefore, the prepared MIPs had a great potential for BB separation.

Objective: HuR (human antigen R)—an RNA-binding protein—is involved in regulating mRNA stability by binding adenylate-uridylate–rich elements. This study explores the role of HuR in the regulation of smooth muscle contraction and blood pressure. Approach and Results: Vascular HuR SMKO (smooth muscle–specific HuR knockout) mice were generated by crossbreeding HuR flox/flox mice with α-SMA (α-smooth muscle actin)-Cre mice. As compared with CTR (control) mice, HuR SMKO mice showed hypertension and cardiac hypertrophy. HuR levels were decreased in aortas from hypertensive patients and SHRs (spontaneously hypertensive rats), and overexpression of HuR could lower the blood pressure of SHRs. Contractile response to vasoconstrictors was increased in mesenteric artery segments isolated from HuR SMKO mice. The functional abnormalities in HuR SMKO mice were attributed to decreased mRNA and protein levels of RGS (regulator of G-protein signaling) protein(s) RGS2, RGS4, and RGS5, which resulted in increased intracellular calcium increase. Consistently, the degree of intracellular calcium ion increase in HuR-deficient smooth muscle cells was reduced by overexpression of RGS2, RGS4, or RGS5. Finally, administration of RGS2 and RGS5 reversed the elevated blood pressure in HuR SMKO mice. Conclusions: Our findings indicate that HuR regulates vascular smooth muscle contraction and maintains blood pressure by modulating RGS expression.

Banana (Musa nana Lour.) have the effect of anti-obesity and lipid modulating properties. However, the influences of banana pulp dietary fibers (BP-DF) on metabolic syndrome (MetS) and gut microbiota (GM) are unknown. In this research, we explore a novel strategy for dietary BP-DF on attenuation of lipid metabolic disease, GM disorder, and associated mechanisms in high-fat diet (HFD) mice. BP-DF can strongly suppress on HFD caused body weight and epididymal fat mass gain, and significantly improved serum lipid profiles, liver lipid profiles, and intestinal function. BP-DF also significantly improved fecal short-chain fatty acids formation and fecal ammonia content. BP-DF impacted the intestinal microorganism at all kinds of taxonomic levels by increasing the proportions of beneficial Lactobacillus, Bacteroidales _S24_7_group, and Alloprevotella and decreasing the disease or obesity associated Sutterella, Streptococcaceae, and Erysipelotrichaceae. The experiments show that BP-DF may use as a functional ingredient for preventing obesity, MetS, and intestinal microorganism imbalance. PRACTICAL APPLICATIONS: Obesity result in many metabolic complications, and it poses a great threat to people's health. Nowadays, the introduction of DF may lead to the development of a new strategy in the treatment of obesity and its metabolic syndrome. Our experiments findings show that BP-DF may use as a functional ingredient for preventing obesity, MetS, and intestinal microorganism imbalance. Therefore, BP-DF can be applied to the development and production of functional food, and can also be used as an important food functional ingredient, which can be added to various food, such as bread, noodles, baked goods, cakes, etc., to improve its nutritional value.

Background: Liver plays an important role in lipid and glucose metabolism. Human antigen R (HuR) as an RNA regulator protein participates in many disease processes. Here, we investigated the specific role of HuR in hepatic steatosis and glucose metabolism. Methods: We investigated the level of HuR in liver from mice fed a normal chow diet (NCD) and high fat diet (HFD). Liver specific HuR knockout (HuRLKO) mice were generated and challenged with an HFD. Lipid levels and glucose metabolism index were examined. Findings: HuR was downregulated in livers of HFD-fed mice. HuRLKO mice showed exacerbated HFD-induced hepatic steatosis but improved glucose tolerance as compared with controls. Consistently, HuR inhibited lipid accumulation in hepatocytes. Mechanically, HuR could bind to the mRNA of phosphatase and tensin homology deleted on chromosome 10 (PTEN), thus increasing their stability and translation. Finally, PTEN over-expression alleviated HFD-induced hepatic steatosis in HuRLKO mice. Interpretation: HuR modulates lipid and glucose metabolism through regulating PTEN expression. Funding Statement: This work was supported by the National Natural Science Foundation of China (no. 81970198, 81770473) and the Taishan Scholar Project of Shandong Province of China (no. tsqn20161066). Declaration of Interests: The authors declare no conflicts of interest. Ethics Approval Statement: The animal experiment was approved by the Animal Care Committee of Shandong University and was performed in compliance with the Animal Management Rules of the Chinese Ministry of Health (Document No. 55, 2001). All procedures conformed to the guidelines from the NIH Guide for the Care and Use of Laboratory Animals.

A growing body of literature has discussed the effect of service trade liberalization on external financial constraints of enterprises in China. Based on a dataset of Chinese manufacturing firms and service trade restriction indices, we contribute to this discussion by analyzing how service trade liberalization affects financial constraints of Chinese enterprises through altering their ability to access external financing, as well as internal financing and commercial credit financing. We find that service trade liberalization alleviates financial constraints of Chinese enterprises through strengthening their ability to obtain both external and internal financing. There exist heterogeneous outcomes for different types of firms. While service trade liberalization eases exporters’ financial constraints, it deteriorates financial constraints of non-exporters through tightening their commercial credit financing. In addition, non-SOEs or SMEs gain from service trade liberalization through easing internal and external financing constraints while SOEs or large enterprises lose due to tightening commercial credit financing constraints.

The purpose of this study was to investigate the effect of okra polysaccharides with similar molecular weight on intestinal ecology. After extraction, ethanol precipitation, deproteinization and purification, two polysaccharide fractions, named as water-extracted polysaccharides (WEP) and alkaline-extracted polysaccharides (AEP), were obtained. According to the results of high performance liquid chromatography, AEP contained mannose, rhamnose, galacturonic acid and galactose, while WEP was composed of galactose, rhamnose, fructose, arabinose, glucose, galacturonic acid and mannose. The compositions of intestinal microbiota were affected by the change of daily substrates. Meanwhile, the concentration of ammonia and gastrointestinal transit time decreased, the total short chain fatty acid (SCFA) levels, moisture of caecum content increased. These findings suggested that the effect of okra polysaccharides on intestinal microbiota was achieved by the output of detrimental compounds and the supply of nutrients. Besides, AEP supported a more pivotal role in manipulating gut microbiota than WEP.

Background: Macamides, the main active components contained in maca, have attracted increasing attention due to their various bioactivities. In this study, crude macamide extract (CME) and purified macamide extract (PME) were prepared by enzyme-assisted extraction and macroporous resin separation, and the anti-fatigue effects of CME and PME were evaluated in a forced swimming model.; Results: The composition analysis results revealed that both CME and PME mainly contain eight kinds of macamide. Based on the results of a weight-loaded forced swimming test, compared with a control group, CME and and PME groups could prolong exhaustive swimming time, increase levels of liver glycogen (LG) and muscle glycogen (MG), accelerate fatty acid oxidation in serum to provide energy, eliminate the accumulation of blood lactic acid (BLA) and blood urea nitrogen (BUN), and decrease the serum biomarkers for muscle damage, such as lactate dehydrogenase (LDH) and creatine kinase (CK). Histological analysis also indicated that CME and PME attenuated damage to skeletal muscle and the myocardium in mice during exercise.; Conclusion: Two macamide extracts have a beneficial effect on relieving physical fatigue by attenuating the damage of skeletal muscle and myocardium during exercise, and a better effect was observed in the PME group. © 2018 Society of Chemical Industry.; © 2018 Society of Chemical Industry.

Smoking is a major preventable risk factor for atherosclerosis. However, the causative link between cigarette smoke and atherosclerosis remains to be established. The objective of this study is to characterize the role of GTP cyclohydrolase 1 (GTPCH1), the rate‐limiting enzyme for de novo tetrahydrobiopterin (BH4) synthesis, in the smoking‐accelerated atherosclerosis and the mechanism involved. In vitro, human umbilical vein endothelial cells were treated with nicotine, a major component of cigarette smoke, which reduced the mRNA and protein levels of GTPCH1 and led to endothelial dysfunction. GTPCH1 overexpression or sepiapterin could attenuate nicotine‐reduced nitric oxide and ‐increased reactive oxygen species levels. Mechanistically, human antigen R (HuR) bound with the adenylateuridylate‐rich elements of the GTPCH1 3′ untranslated region and increased its stability; nicotine inhibited HuR translocation from the nucleus to cytosol, which downregulated GTPCH1. In vivo, nicotine induced endothelial dysfunction and promoted atherosclerosis in ApoE−/− mice, which were attenuated by GTPCH1 overexpression or BH4 supplement. Our findings may provide a novel and promising approach to atherosclerosis treatment.

We used the University of Washington Quality of Life (UW-QoL) questionnaire (version 4) to assess the long-term quality of life (QoL) of patients with oral squamous cell carcinoma treated with or without reconstruction with a microvascular free flap, and all patients with T2-T4 oral squamous cell carcinoma (SCC) treated in this way were eligible for the study. A total of 139 patients' personal details, medical history, and QoL scores were collected and analysed. The mean (SD) overall QoL score was 73.09 (14.8) for patients with T2-T4 oral SCC. The mean (SD) global QoL scores of patients who had had reconstructions was 75.68 (13.85) and of patients who had not 71.00 (15.34) (t=1.864, df=137, p=0.064). Univariate and multivariate analyses indicated that site of tumour, T stage, and the need for postoperative radiotherapy had significant effects on the global QoL scores. Among patients with T2 oral SCC there was no significant difference between patients who did and did not have reconstructions. However, there were significant differences among patients with T3/T4 SCC in scores for appearance, recreation, mood, anxiety, chewing, swallowing, and speech depending on whether they did or did not have reconstructions. There were significant differences in the domains recreation, chewing, speech, and taste, depending on the primary site of the tumour. We conclude that reconstruction with a microvascular free flap had a beneficial effect on the treatment of SCC and improved the QoL of patients with oral SCC.

Cooking properties and retrogradation process of black rice (BR) with high hydrostatic pressure (HHP, 200–400 MPa/15 min) soaking were evaluated in this study. Results showed that the water absorption capacity of BR with HHP soaking was higher than that of control one. The HHP soaking also generated the lower leached amylose and hardness, while the higher springiness, cohesiveness, and resilience for cooked BR. In addition, the DSC data revealed that the enthalpy of cooked BR was influenced by HHP treatment, and the degree of retrogradation was decreed from 91% to 71% (storage for 21 days). Furthermore, the analysis of X-ray diffraction (XRD) revealed that the HHP treatment weakened the intensity of the peak of the 14 day-retrograded samples and decreased the relative crystallinity from 19.92% to 15.94% (storage for 21 days). These findings suggest that the HHP soaking could be a potentially applicable pretreatment for producing cooked BR with high quality.

Recently, polymeric micelles with different biodegradable hydrophobic cores, such as poly(lactide-co-glycolide) (PLGA) and poly(lactide-co-3(S)-methyl-morpholine-2,5-dione) (PLMD), have been used f...

Studies on how education shapes adult health have largely conceptualized education as the quantity of schooling attained, coined the "vertical dimension" of education. While this dimension is important, heterogeneity within levels of education (the "horizontal dimension") may also shape health. Using data from the 2010-2014 American Community Survey on adults aged 45-64 with a Bachelor's degree (N = 667,362), we investigate the association between a key indicator of adult health (physical functioning) and an understudied horizontal dimension of education (college major). We find that physical functioning in midlife varies significantly by college major. For instance, the odds of poor functioning for men who majored in Psychology/Social Work were 1.9 (95% CI: 1.7, 2.1) times greater than for men who majored in Business. However, all college graduates, regardless of major, report better functioning than non-graduates. We also find that inequalities in midlife functioning across majors largely reflect differences in human capital skills and financial returns in the labor market. Taken together our findings suggest that college major is an important component of health stratification and should be integrated into the literature on health inequalities.

Recently, the red blood cell (RBC) membrane has been used as a mimetic nanocoating for nanoparticles for drug delivery systems to promote their biocompatibility. In the present study, the nano-sized RBC membrane was coated on the surface of gene complexes through electrostatic interactions to prepare biomimetic gene delivery systems so as to improve their biocompatibility and prolong their circulation time in vivo. The structure of the biomimetic gene delivery systems was determined by transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM). They exhibited low cytotoxicity and high transfection efficiency in endothelial cells (ECs), which could improve the migration ability of ECs. Besides, the biomimetic gene delivery systems exhibited strong immune evasion and long in vivo circulation time. The phagocytic rate of these biomimetic gene delivery systems reduced 52% compared with that of the PLGA-PEI/pZNF580 control group (without RBC membrane modification). Their circulation time in vivo was more than 2 times higher than that of the control group. Consequently, we provide a simple method for the preparation of camouflaged gene delivery systems, which can further facilitate the development of a gene delivery platform for the therapy of vascular diseases via enhancing EC transfection. This strategy will open up a new avenue for gene delivery systems by RBC membrane camouflage.

Inhibiting vascular restenosis remains a tricky challenge for the postoperative development of cardiovascular interventional therapy. The ideal approaches should activate endothelial cells (ECs) and restrain smooth muscle cells (SMCs), however, they are commonly contradictory. Herein, a strategy was developed for synchronizing ECs promotion and SMCs inhibition by codelivery DNA and siRNA for combination therapy. Thus, an easy and efficient strategy integrated dual-superiorities of precise targeting and dual therapeutic genes to precisely regulate the behaviors of ECs and SMCs. The ECs-targeting REDV peptide and SMCs-targeting VAPG peptide grafted anionic polymers were used to surface-functionalize the delivery nanoplatforms for vascular endothelial growth factor (VEGF) plasmids and ERK2 siRNA delivery, respectively. The dual targeting-nanoparticles were prepared by physical mixing method, and their outstanding advantages were confirmed by the co-culture experiments. In comparison with single targeting-nanoparticles and dual non-targeting-nanoparticles, the dual targeting-nanoparticles simultaneously enhanced ECs proliferation/migration and restrained SMCs proliferation/migration. Moreover, the dual targeting-nanoparticles group manifested the highest VEGF expression in ECs and the lowest ERK2 expression in SMCs. In summary, the two-pronged strategy with dual targeting-nanoparticles provides a valuable cornerstone for synchronizing ECs promotion and SMCs inhibition.

Ischemic injury is mostly caused by insufficient blood supply, excessive inflammation and increased oxidative stress. Pro-angiogenic gene delivery can foster vascular remodeling and increase blood perfusion, but the adverse ischemia microenvironment leads to inadequate therapy effect and high recurrence rate. A novel combination strategy should be developed for the effective therapy of ischemic injury via remediating microenvironment and accelerating revascularization simultaneously. Carbon monoxide (CO) exhibits multiple bio-functions, especially anti-inflammation and anti-oxidant effects, suggesting that it might be an attractive candidate for ischemia therapy. Herein, for the first time, “controlled CO release” was introduced to treat limb ischemia. We developed the controlled CO delivery nanoplatform (PR@CORM) via encapsulating CO-releasing molecule-401 by the stimulus-responsive and ECs-adhesive peptide functionalized nanocarrier. In response to oxidants, PR@CORM can intelligently release therapeutic CO gas to remediate the microenvironment, but the pro-angiogenic effect is still disappointing. We thereby combined “CO therapy” and “pro-angiogenic gene therapy” to achieve the cooperative effects for ischemia therapy. This strategy exhibits superiority in the treatment of ischemic injury through extensively normalizing the microenvironment and accelerating revascularization. The “controlled CO release” serves as a scavenger of inflammation and reactive oxygen species, and pro-angiogenic gene holds potential for ischemia therapy.

Multi-temporal remote sensing image registration is a geometric symmetry process that involves matching a source image with a target image. To improve the accuracy and enhance the robustness of the algorithm, this study proposes an end-to-end registration network—a bidirectional symmetry network based on dual-field cyclic attention for multi-temporal remote sensing image registration, which mainly improves feature extraction and feature matching. (1) We propose a feature extraction framework combining an attention module and a pre-training model, which can accurately locate important areas in images and quickly extract features. Not only is the dual receptive field module designed to enhance attention in the spatial region, a loop structure is also used to improve the network model and improve overall accuracy. (2) Matching has not only directivity but also symmetry. We design a symmetric network of two-way matching to reduce the registration deviation caused by one-way matching and use a Pearson correlation method to improve the cross-correlation matching and enhance the robustness of the matching relation. In contrast with two traditional methods and three deep learning-based algorithms, the proposed approach works well under five indicators in three public multi-temporal datasets. Notably, in the case of the Aerial Image Dataset, the accuracy of the proposed method is improved by 39.8% compared with the Two-stream Ensemble method under a PCK (Percentage of Correct Keypoints) index of 0.05. When the PCK index is 0.03, accuracy increases by 46.8%, and increases by 18.7% under a PCK index of 0.01. Additionally, when adding the innovation points in feature extraction into the basic network CNNGeo (Convolutional Neural Network Architecture for Geometric Matching), accuracy is increased by 36.7% under 0.05 PCK, 18.2% under 0.03 PCK, and 8.4% under 0.01 PCK. Meanwhile, by adding the innovation points in feature matching into CNNGeo, accuracy is improved by 16.4% under 0.05 PCK, 9.1% under 0.03 PCK, and 5.2% under 0.01 PCK. In most cases, this paper reports high registration accuracy and efficiency for multi-temporal remote sensing image registration.

Gene-engineered vascular endothelial cells have acquired satisfactory ischemia therapy in animal models. However, its clinical process is still severely limited by the hurdle of transfection-structure-toxicity and single therapeutic pathway. To this end, inspired by green process, we innovatively introduced “green” concept into gene delivery. “Green” means fully utilizing carrier and gene via simple, economic and nontoxic process. Herein, the combined strategy of “bioactive carrier” and “CO2-modification” was proposed. On the basis of the cascaded expression from ZNF580 to endothelial nitric oxide synthase (eNOS), we designed cationic peptide based on L-arginine as bioactive carrier to deliver pZNF580. Furthermore, CO2-bubbling was performed to endow bioactive carrier with pH-sensitive nanobomb effect. The carrier dramatically enhanced endo/lysosomal escape via nanobomb effect for high pZNF580 delivery and expression. In turn, carrier changed “enhanced vehicle” identity into NO substrate after completing gene delivery and further was catalyzed by transfection-triggered eNOS overexpression, which greatly amplified NO generation via their synergistic promotion. This simple design obtained dual-high results of ZNF580 (to mediate angiogenesis) and NO (to mediate angiogenesis and anti-inflammation) level. In critical hind limb ischemia (CLI) mice, the strategy simultaneously promoted angiogenesis and mitigated inflammation, which co-worked to recover blood flow and rescue limb. Besides, it was nontoxic to muscle tissue. The multi-pathways achieved significant CLI treatment. Taken together, we established a positive feedback between CO2-enhanced bioactive carrier and target-gene expression via green design and process. This can maximize and expand the output of gene therapy through simplifying the input of gene complexes, thus guaranteeing high transfection, simple structure and low toxicity. We believe that this green strategy will provide a novel viewpoint for gene delivery.

Background Bi-potential hepatic progenitor cells can give rise to both hepatocytes and cholangiocytes, which is the last phase and critical juncture in terms of sequentially hepatic lineage restriction from any kind of stem cells. If their differentiation can be controlled, it might access to functional hepatocytes to develop pharmaceutical and biotechnology industries as well as cell therapies for end-stage liver diseases. Methods In this study, we investigated the influence of Cx32 and Cx43 on hepatocyte differentiation of WB-F344 cells by in vitro gain and loss of function analyses. An inhibitor of Cx32 was also used to make further clarification. To reveal p38 MAPK pathway is closely related to Cxs, rats with 70% partial hepatectomy were injected intraperitoneally with a p38 inhibitor, SB203580. Besides, the effects of p38 MAPK pathway on differentiation of hepatoblasts isolated from fetal rat livers were evaluated by addition of SB203580 in culture medium. Results In vitro gain and loss of function analyses showed overexpression of Connexin 32 and knockdown of Connexin 43 promoted hepatocytes differentiation from hepatic progenitor cells. In addition, in vitro and ex vivo research revealed inhibition of p38 mitogen-activated protein kinase pathway can improve hepatocytes differentiation correlating with upregulation of Connexin 32 expression and downregulation of Connexin 43 expression. Conclusions Here we demonstrate that Connexins play crucial roles in facilitating differentiation of hepatic progenitors. Our work further implicates that regulators of Connexins and their related pathways might provide new insights to improve lineage restriction of stem cells to mature hepatocytes. Electronic supplementary material The online version of this article (doi:10.1186/s13287-017-0703-2) contains supplementary material, which is available to authorized users.

Recently, gene therapy has attracted much attention, especially for the treatment of vascular disease. However, it is still challenging to develop the gene carriers with high biocompatibility as well as highly efficient gene delivery to overcome multiple barriers. Herein, a frequently used cell-penetrating peptide PKKKRKV (TAT) was selected as a functional sequence of the gene carrier with distinctive cell-penetrating ability. REDV peptide with selectively targeting function for endothelial cells (ECs) and nuclear localization signals (NLS) were integrated with this TAT peptide to obtain a highly efficient gene delivery system with ECs specificity and nucleus accumulation capacity. Besides, the glycine sequences with different repeat numbers were inserted into the above integrated peptide. These glycine sequences acted as a flexible spacer arm to exert the targeting, cell-penetrating, and nucleus accumulation functions of each functional peptide. Three tandem peptides REDV-Gm-TAT-Gm-NLS (m = 0, 1, and 4) ...

Due to its hollow structure, halloysite has been considered for owning the potential of application in preparing low dielectric constant material. However its polarity has suppressed such potential and raised the dielectric constant of its polymeric composite. In this study, the non-polar carbon intercalated halloysite (C-HNT) was successfully prepared via in-situ polymerization and thermal treatments. Characterizations of the carbon intercalated halloysite confirmed that acrylonitrile and acrylamide as monomers had been intercalated and co-polymerized between the halloysite layers. The copolymer carbonized at 600°C under nitrogen atmosphere with the lumen of halloysite being preserved and water being eliminated. A series of carbon intercalated halloysites/organosilicone hybrid films were prepared via sol-gel process, and their hydrophobicity, thermal property, mechanical property and dielectric property were characterized. The results showed that the present of carbon intercalated halloysite strengthened the hydrophobicity, thermal property, mechanical property of the hybrid film. The lowest dielectric constant of the hybrid film samples was 2.46 at 1MHz. It reveals that the intercalated process suppresses the polarity of halloysite and revives halloysite's great potential for being used as low dielectric constant material. Keywords: Halloysite, Intercalation, Crystalline structure, Low dielectric constant, Organosilicone hybrid film

Purpose: Radiomics has shown great promise to use quantifiable imaging characteristics to predict the behavior and prognosis of neoplasms. This is the first study to evaluate whether radiomic texture analysis can predict outcomes in gastroesophageal junction adenocarcinoma (GEJAC) treated with neoadjuvant chemoradiotherapy (CRT). Materials and Methods: Pretreatment contrast-enhanced CT images of 146 patients with stage II-III GEJAC were reviewed (2009–2011), and randomly split into training and validation groups at a 1:1 ratio stratified with baseline clinical characteristics. Whole-tumor texture was assessed using quantitative image features based on intensity, shape, and gray-level co-occurrence matrix. The relevant pretreatment texture features, in addition to the significant baseline clinical features to predict survival, were identified using multivariate Cox proportional hazard regression model with stepwise variable selection in the training sample and verified in the validation sample, to facilitate the proposal of a multi-point index for standard patient pre-treatment risk classification. Results: Of the factors identified in the training cohort independently associated with OS, only shape compactness (p = 0.04) and pathologic grade differentiation (PDG) (p = 0.02) were confirmed in the validation sample. Using both parameters, we created a 3-point risk classification index: low-risk (well-moderate PDG and high compactness), medium-risk (poor PDG or low compactness), and high-risk (poor PDG and low compactness). The risk index showed a strong negative association with postoperative pathologic complete response (pCR) (p = 0.04). Median OS for the high-, medium-, and low-risk groups were 23, 51, and ≥72 months, respectively (p

// Xue Li 1, 2, * , Lujun Zhao 1, * , Wencheng Zhang 1 , Chengwen Yang 1 , Zhen Lian 1 , Shuai Wang 1 , Ningbo Liu 1 , Qingsong Pang 1 , Ping Wang 1 and Jinming Yu 1, 2 1 Department of Radiation Oncology and Key Laboratory of Cancer Prevention and Therapy, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin 300060, China 2 Department of Radiation Oncology, Shandong University Affiliated Shandong Cancer Hospital and Institute, Jinan 250000, China * These authors have contributed equally to this work Correspondence to: Jinming Yu, email: sdyujinming@163.com Keywords: esophageal squamous cell carcinoma, supraclavicular nodes, celiac nodes, common hepatic nodes, radiotherapy Received: February 17, 2017 Accepted: April 17, 2017 Published: May 19, 2017 ABSTRACT The purpose of this study is to assess the prognostic value of supraclavicular nodes, left gastric nodes, celiac nodes and common hepatic nodes metastasis in esophageal squamous cell carcinoma (ESCC) treated with definitive radiotherapy. A total of 293 ESCC patients treated with radiotherapy or chemoradiotherapy entered the study. The results showed that the presence of supraclavicular nodes (χ 2 = 0.075, P = 0.785) and left gastric nodes (χ 2 = 3.603, P = 0.058) metastasis had no significant influence on survival, while celiac nodes (χ 2 = 33.775, P < 0.001) and common hepatic nodes (χ 2 = 42.350, P < 0.001) metastasis were associated with significantly shorter survival, regardless of the sites of primary tumor. Multivariate analysis showed that celiac nodes (HR: 0.457, 95% CI: 0.256-0.816; P = 0.008) and common hepatic nodes (HR: 0.241, 95% CI: 0.092-0.630; P = 0.004) metastasis were independently adverse indicator of survival in upper ESCC. While in the middle and lower ESCC, only the common hepatic nodes (middle ESCC: HR: 0.345, 95% CI: 0.161-0.738, P = 0.006; lower ESCC: HR: 0.377, 95% CI: 0.160-0.890, P = 0.026) metastasis was an independently adverse indicator of survival. In conclusion, our study demonstrated that in ESCC treated with definitive radiotherapy, both of celiac nodes and common hepatic nodes metastasis were adverse indicator of survival in upper ESCC, and common hepatic nodes metastasis were adverse indicator of survival in middle and lower ESCC. Supraclavicular nodes an left gastric nodes metastasis is not associated with patients survival in ESCC.

An efficient preparation procedure has been proposed for the synthesis of 5,6-di-O-cinnamyl-l-ascorbic acid ester (5,6-CA–AA) and 2-O-cinnamyl-l-ascorbic acid ester (2-CA–AA) via ascorbic acid (AA) and cinnamic acid (CA), and their anti-oxidant and antibacterial activities were also investigated. Among the factors affecting the synthesis efficiency of 5,6-CA–AA and 2-CA–AA, reaction solvent and substrate molar ratio were very important, resulting in 76 and 54% total yield, respectively. Further studies indicated that 5,6-CA–AA maintained the inherent inhibitory ability of AA in α, α-diphenyl-β-picrylhydrazyl radical scavenging assay. Also, it had stronger scavenging activity against hydroxyl radicals than that of standard tert-butylhydroquinone (EC50: 7.84 vs. 8.38 mM). In addition, 5,6-CA–AA and 2-CA–AA possessed comparable antibacterial activity to that of CA. Significantly, the application of ascorbyl cinnamate could retard the lipid oxidation of peanut oil and pork, and the corresponding perioxide value and thiobarbituric acid reactive substance content decreased by 32.0 and 56.5%, respectively. Moreover, the microbial reproduction of the pork treated with mixed ascorbyl cinnamate was attenuated to some extent. These investigations will pave the way for designing more efficient bifunctional derivatives of AA and CA.

// Yun Zhang 1, 2, * , Jing Liu 3, * , Wencheng Zhang 4 , Weiye Deng 5, 6 , Jinbo Yue 2 1 School of Medicine and Life Sciences, University of Jinan Shandong Academy of Medical Sciences, Jinan, Shandong, China 2 Department of Radiation Oncology, Shandong Cancer Hospital and Institute, Shandong Cancer Hospital Affiliated to Shandong University, Jinan, Shandong, China 3 Graduate Education Center, Shandong Academy of Medical Sciences, Jinan, Shandong, China 4 Tianjin Cancer Hospital, Tianjin, China 5 Department of Radiation Oncology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA 6 Division of Epidemiology, Human Genetics and Environmental Sciences, The University of Texas School of Public Health at Houston, Houston, Texas, USA * These authors have contributed equally to this work Correspondence to: Jinbo Yue, email: yuejinbo@hotmail.com Keywords: esophageal cancer, radiation therapy, radiation oncologist, questionnaire Received: February 08, 2017 Accepted: March 28, 2017 Published: April 05, 2017 ABSTRACT Lots of controversies were found about the treatment in relation to radiation therapy (RT) for esophageal squamous cell carcinoma (ESCC). We designed a questionnaire of these controversies to do a pan-Chinese survey of radiation oncologists (ROs). For operable ESCC, 53% ROs chose surgery plus postoperative chemoradiotherapy (CRT), while 40% chose preoperative CRT plus surgery. For target volume of postoperative RT, most ROs (92%) would delineate tumor bed plus involved lymph nodes region before surgery. For definitive RT, most ROs (81%) would give patients higher RT dose to 60–65Gy. For radiation target volume, most ROs would give patients prophylactic irradiation of the bilateral superclavicular-lymph nodes region for cervical ESCC (93%), and the left gastric lymph nodes region for lower thoracic ESCC (72%). For the treatment of mediastinal lymph nodes, 72% ROs preferred elective nodal irradiation, while 28% did the involved nodal irradiation. For concurrent chemotherapy regimen, PF (5-Fu + cisplatin) and TP (cisplatin + paclitaxel) were used widely (49% and 46%, respectively). During simulation, four-dimensional computer tomography (4D CT) was not widely used (48%), even for cervical or lower thoracic ESCC (52%). For daily RT delivery, only 66% ROs would perform imaging guidance RT daily. In summary, more controversies existed in the treatment of ESCC with RT in China, including treatment strategy, radiation dose and target contour. Future goals include standardization of treatment strategy, radiation dose, and target contour, and application of 4D CT and daily imaging guidance, and pursuit of randomized trials in Chinese population.

Enzyme-assisted extraction (EAE) coupled with macroporous resin column chromatography (MRCC) was successfully used for the extraction and purification of cordycepin and adenosine from cultured Cordyceps militaris. After optimized by the response surface methodology (RSM), the extraction yields of cordycepin and adenosine were 86.45% and 70.06%, respectively. Subsequently, under the optimal separation conditions on NKA-II resins (loading 1 BV of extraction solution with pH 8.0, and eluting with 4 BVs of 70% ethanol at a flow rate of 3 BV/h), cordycepin (purity of 32.5%), and adenosine (purity of 39.9%) were obtained with an overall recovery rate more than 90%.

Liver kinase B1 (LKB1) is a serine/threonine protein kinase ubiquitously expressed in mammalian cells. It was first identified in Peutz-Jeghers syndrome as a tumor suppressor gene. Whether endothelial LKB1 regulates angiogenesis and tumor growth is unknown. In this study, we generated endothelial cell-specific LKB1-knockout (LKB1endo-/-) mice by crossbreeding vascular endothelial-cadherin-Cre mice with LKB1flox/flox mice. Vascular endothelial growth factor (VEGF) level was highly co-stained in endothelial cells but not in macrophages in LKB1endo-/- mice. Consistently, LKB1endo-/- mouse tissues including the lung, skin, kidney and liver showed increased vascular permeability. Tumors implanted in LKB1endo-/- mice but not macrophage-specific LKB1-knockout mice grew faster and showed enhanced vascular permeability and increased angiogenesis as compared with those implanted in wild-type mice. Injection of VEGF-neutralizing antibody but not the isotype-matched control antibody decreased endothelial-cell angiogenesis and tumor growth in vivo. Furthermore, LKB1 deletion enhanced mouse retinal and cell angiogenesis, and knockdown of VEGF by small-interfering RNA decreased endothelial cell proliferation and migration. Re-expression of LKB1 or knockdown of VEGF receptor 2 decreased the overproliferation and -migration observed in LKB1endo-/- cells. Mechanistically, LKB1 could bind to the VEGF transcription factor, specificity protein 1 (Sp1), which then inhibited the binding of Sp1 to the VEGF promoter to reduce VEGF expression. Endothelial LKB1 may regulate endothelial angiogenesis and tumor growth by modulating Sp1-mediated VEGF expression.

// Jing Kong 1,* , Ya Zhang 1,* , Shanshan Liu 1 , Hongxuan Li 1 , Shangming Liu 2 , Jingjing Wang 3 , Xiaoteng Qin 1 , Xiuxin Jiang 4 , Jianmin Yang 1 , Cheng Zhang 1 and Wencheng Zhang 1 1 The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Qilu Hospital of Shandong University, Jinan, Shandong, China 2 Key Laboratory of the Ministry of Education for Experimental Teratology, Department of Histology and Embryology, Shandong University School of Medicine, Jinan, China 3 Institute of Pathology and Pathophysiology, Shandong University School of Medicine, Jinan, China 4 Department of General Surgery, Virtual Laboratory, Qilu Hospital of Shandong University, Jinan, China * These authors have contributted equally to this work Correspondence to: Wencheng Zhang, email: // Keywords : abdominal aortic aneurysm, melatonin, HuR, MMP2, MMP9, Pathology Section Received : December 16, 2016 Accepted : January 24, 2017 Published : February 04, 2017 Abstract Abdominal aortic aneurysm (AAA) affects more than 5% of the population in developed countries and the pharmacotherapies for AAA are limited. Here, we explored whether melatonin regulates the development of AAA. In smooth muscle cells, melatonin treatment decreases angiotensin II-induced matrix metalloproteinase 2 (MMP2) and MMP9 expression. Human antigen R (HuR) could bind with the adenylateuridylate-rich elements of MMP2 and MMP9 mRNAs 3′ untranslated region, resulting in the increased stability of MMP2 and MMP9 mRNAs. HuR is required for angiotensin II-induced MMP2 and MMP9 expression. Moreover, melatonin suppresses angiotensin II-induced HuR expression through inhibiting NF-κB signaling, leading to decreased MMP2 and MMP9 levels. Finally, melatonin attenuates the development of AAA in ApoE -/- mice infused with angiotensin II in vivo . These data support a role of HuR in the development of AAA and possible therapeutic roles for melatonin and/or HuR inhibition in AAA.

Gene therapy is a promising strategy for angiogenesis, but developing gene carriers with low cytotoxicity and high gene delivery efficiency in vivo is a key issue. In the present study, we synthesized the CAGW peptide- and poly(ethylene glycol) (PEG)-modified amphiphilic copolymers. CAGW peptide serves as a targeting ligand for endothelial cells (ECs). Different amounts of CAGW peptide were effectively conjugated to the amphiphilic copolymer via heterofunctional poly(ethylene glycol). These CAG- and PEG-modified copolymers could form nanoparticles (NPs) by self-assembly method and were used as gene carriers for the pEGFP-ZNF580 (pZNF580) plasmid. CAGW and PEG modification coordinately improved the hemocompatibility and cytocompatibility of NPs. The results of cellular uptake showed significantly enhanced internalization efficiency of pZNF580 after CAGW modification. Gene expression at mRNA and protein levels demonstrated that EC-targeted NPs possessed high gene delivery efficiency, especially the NPs with higher content of CAGW peptide (1.16 wt %). Furthermore, in vitro and in vivo vascularization assays also showed outstanding vascularization ability of human umbilical vein endothelial cells treated by the NP/pZNF580 complexes. This study demonstrates that the CAGW peptide-modified NP is a promising candidate for gene therapy in angiogenesis.

If plasmid complexes prepared from targeting carriers have endothelial cell selectivity and high transfection efficiency, they can specifically enhance rapid endothelialization by delivering the corresponding gene plasmids. A high content of functional groups in the carriers benefits high selectivity efficiency. Herein, we have synthesized a comb-shaped polymer bearing several Arg-Glu-Asp-Val (REDV) peptides and poly(ethylene glycol) as a pEGFP-ZNF580 gene carrier with cell-type recognition of human endothelial cells. An amphiphilic block copolymer of poly(2-hydroxyethyl methacrylate)-block-poly(ε-caprolactone)-graft-poly(ethylene glycol)-graft-poly(ethyleneimine) conjugated with REDV peptide (PHEMA-b-PCL-g-PEG-g-PEI-REDV) is synthesized, and nanoparticles of it are prepared by polymer self-assembly. This polycationic PHEMA-b-PCL-g-PEG-g-PEI-REDV carrier effectively condenses pEGFP-ZNF580 plasmid to form REDV-targeted complexes and protects pEGFP-ZNF580 integrity from enzymatic hydrolysis. These REDV-targeted complexes exhibit low cytotoxicity but high transfection efficiency to EA.hy926 cells compared with non-targeted complexes (PHEMA-b-PCL-g-PEG-g-PEI/pEGFP-ZNF580) as demonstrated by MTT assay, fluorescence-activated cell sorting analysis and fluorescence analysis. Furthermore, the relative protein level of endothelial cells transfected by REDV-targeted complexes is higher than that by non-targeted complexes. Therefore, REDV-bearing carriers may have potential as effective and targeting transfer carriers for pEGFP-ZNF580 plasmid, and their complexes can be used in the endothelialization of artificial vascular grafts.

In this work, a biodegradable star-shaped copolymer poly(lactide-co-3(S)-methyl-morpholine-2,5-dione)6 (Star-(PLMD)6) was synthesized via ring-opening polymerization (ROP), and subsequently a gene carrier Star-PLMD-g-PEI-g-PEG-CREDVW was prepared by grafting polyethyleneimine (PEI), polyethylene glycol (PEG) and targeting peptide REDV onto Star-(PLMD)6. This gene carrier could form stable micelles to condense pEGFP-ZNF580 through electrostatic interaction. The resulting complexes were biocompatible and showed high efficiency in gene delivery. In addition, these complexes exhibited high selectivity for endothelial cells (ECs), high transfection efficiency and enhanced migration of ECs. The protein level of ZNF580 expression was significantly high (up to 85%), while the control group was only 51%. This combination of degradability, targeting ligand and star-structure strategy exhibits a significant advantage in transfection efficiency and migration of ECs.