Search

Objective: The goal of the present study was to determine the expression of yes-associated protein 1 (YAP1) in renal tissues of mice with lupus nephritis (LN) and elucidate its role in the progression of renal fibrosis. Methods: C57BL/6 mice and MRL/lpr mice were selected for experimental comparison. Mouse kidney tissues were removed and sectioned for hematoxylin and eosin staining, Masson’s trichome staining, Sirius staining, and immunohistochemistry. The mRNA and protein levels of YAP1 in mouse kidney tissues were detected, and the correlation between YAP1 and fibronectin (FN) mRNA levels was analyzed. Mouse renal epithelial cells were used for in vitro experiments. After transfection and stimulation, the cells were divided into 4 groups, namely the C57BL/6 serum group (group 1), the MRL/lpr serum group (group 2), the MRL/lpr serum + siRNA-negative control group (group 3), and the MRL/lpr serum + siRNA-YAP1 group (group 4). Epithelial-mesenchymal transition (EMT) markers in each group were detected by Western blotting and immunofluorescence staining. Serum creatinine, blood urea nitrogen, and urinary protein levels were detected and assessed for their correlation with YAP1 mRNA levels by Spearman’s analysis. Results: Compared to C57BL/6 mice, MRL/lpr mice exhibited obvious changes in fibrosis in renal tissues. In addition, YAP1 expression was significantly higher in the renal tissues of MRL/lpr mice than in those of C57BL/6 mice, and YAP1 mRNA levels were positively correlated with those of FN. YAP1 silencing in lupus serum-stimulated cells could effectively relieve serum-induced EMT. Finally, we observed that YAP1 mRNA levels in mouse kidney tissue were significantly and positively correlated with the degree of renal function injury. Conclusion: YAP1 expression in the kidney tissues of LN mice was higher than that observed in normal mice, indicating that YAP1 may play an important role in the occurrence and development of LN.

PIM2 (proviral integration site for Moloney murine leukemia virus 2) kinase plays an important role as an oncogene in multiple cancers, such as leukemia, liver, lung, myeloma, prostate and breast cancers. PIM2 is largely expressed in both leukemia and solid tumors, and it promotes the transcriptional activation of genes involved in cell survival, cell proliferation, and cell-cycle progression. Many tumorigenic signaling molecules have been identified as substrates for PIM2 kinase, and a variety of inhibitors have been developed for its kinase activity, including SMI-4a, SMI-16a, SGI-1776, JP11646 and DHPCC-9. Here, we summarize the signaling pathways involved in PIM2 kinase regulation and PIM2 mechanisms in various neoplastic diseases. We also discuss the current status and future perspectives for the development of PIM2 kinase inhibitors to combat human cancer, and PIM2 will become a therapeutic target in cancers in the future.

Clinical investigations have shown that a nonimmunogenic "cold" tumor is usually accompanied by few immunopositive cells and more immunosuppressive cells in the tumor microenvironment (TME), which is still the bottleneck of immune activation. Here, a fluorine assembly nanocluster was explored to break the shackles of immunosuppression, reawaken the immune system, and turn the cold tumor "hot." Once under laser irradiation, FS@PMPt produces sufficient reactive oxygen species (ROS) to fracture the ROS-sensitive linker, thus releasing the cisplatin conjugated PMPt to penetrate into the tumors and kill the regulatory T cells (Tregs) and myeloid-derived suppressor cells (MDSCs). Meanwhile, ROS will induce potent immunogenic cell death (ICD) and further promote the accumulation of dendritic cells (DCs) and T cells, therefore not only increasing the infiltration of immunopositive cells from the outside but also reducing the immunosuppressive cells from the inside to break through the bottleneck of immune activation. The FS@PMPt nanocluster regulates the immune process in TME from negative to positive, from shallow to deep, to turn the cold tumor into a hot tumor and provoke a robust antitumor immune response.

To determine the efficacy and safety of Endostar combined with gemcitabine and cisplatin in the treatment of NSCLC, and provide evidence-based reference for clinical drug use. Retrieved from Cochrane Library, PubMed, Embase, ClinicalTrials, CNKI, Wanfang and VIP database, randomized controlled trials (RCT) were collected. Pooled standard mean differences (SMD) and 95% confidence intervals (CI) were analyzed using R software with a random-effects model. Data from a total of 27 RCTs were included, involving 1646 patients were analyzed. Results of meta-analysis showed that response rate and clinical benefit rate of trial group were significantly higher than those of control group. There was no statistical significance in the incidence of leucopenia, thrombocytopenia and gastrointestinal reaction between 2 groups. The results showed that Endostar combined with gemcitabine and cisplatin may generally improve therapeutic efficacy of NSCLC patients, without increasing the incidence of adverse reactions.

The southern Red Sea is genetically distinct from the rest of the basin; yet the reasons responsible for this genetic separation remain unclear. Connectivity is a vital process for the exchange of individuals and genes among geographically separated populations, and is necessary for maintaining biodiversity and resilience in coral reef ecosystems. Here, using long-term, high-resolution, 3-D backward particle tracking simulations, we investigate the physical connectivity of coral reefs in the southern Red Sea with neighbouring regions. Overall, the simulation results reveal that the southern Red Sea coral reefs are more physically connected with regions in the Indian Ocean (e.g., the Gulf of Aden) than with the northern part of the basin. The identified connectivity exhibits a distinct monsoon-related seasonality. Though beyond the country boundaries, relatively remote regions of the Indian Ocean may have a substantial impact on the southern Red Sea coral reef regions, and this should be taken into consideration when establishing conservation strategies for these vulnerable biodiversity hot-spots.

There is an urgent need for animal models of COVID-19 to study immunopathogenesis and test therapeutic intervenes. In this study we showed that NSG mice engrafted with human lung (HL) tissue (NSG-L mice) could be infected efficiently by SARS-CoV-2, and that live virus capable of infecting Vero cells was found in the HL grafts and multiple organs from infected NSG-L mice. RNA-seq examination identified a series of differentially expressed genes, which are enriched in viral defense responses, chemotaxis, interferon stimulation, and pulmonary fibrosis between HL grafts from infected and control NSG-L mice. Furthermore, when infecting humanized mice with human immune system (HIS) and autologous HL grafts (HISL mice), the mice had bodyweight loss and hemorrhage and immune cell infiltration in HL grafts, which were not observed in immunodeficient NSG-L mice, indicating the development of anti-viral immune responses in these mice. In support of this possibility, the infected HISL mice showed bodyweight recovery and lack of detectable live virus at the later time. These results demonstrate that NSG-L and HISL mice are susceptible to SARS-CoV-2 infection, offering a useful in vivo model for studying SARS-CoV-2 infection and the associated immune response and immunopathology, and testing anti-SARS-CoV-2 therapies.

Background Endometriosis is a chronic hormonal inflammatory disease characterized by the presence of endometrial tissue outside the uterus. Endometriosis often causes infertility, which brings physical and mental pain to patients and their families. Methods We examined the functions of heat shock factor 1 (HSF1) in endometriosis development through cell count assay, cell-scratch assay and clone formation experiments. We used quantitative real-time PCR (qRT-PCR) and Western blot (WB) to detect HSF1 expression. Glucose and lactate levels were determined using a glucose (GO) assay kit and a lactate assay kit. Furthermore, we used a HSF1 inhibitor-KRIBB11 to establish a mouse model of endometriosis. Results Our data demonstrated that HSF1 promoted endometriosis development. Interestingly, HSF1 enhanced glycolysis via up-regulating PFKFB3 expression in endometriosis cells, which was a key glycolysis enzyme. Consistently, the HSF1 inhibitor KRIBB11 could abrogate endometriosis progression in vivo and in vitro. Conclusions Findings indicate that HSF1 plays an important role in endometriosis development, which might become a new target for the treatment of endometriosis. Electronic supplementary material Supplementary data are available.

Non-suicidal self-injury (NSSI) refers to any intentional, self-inflicted behavior that causes direct damage to body tissues (Kerr et al., 2010), and has emerged as a challenging public health issue worldwide, especially among adolescents. The most common presentations of NSSI include skin-cutting, severe scratching, and burning (Whitlock et al., 2006). In a community-based study conducted in England, 6% of 5506 adolescents reported self-harm in 2014, and 1320 adolescents (74% aged 15‒17 years) presented to hospitals due to non-fatal self-harm (Geulayov et al., 2018). In Australia, 8% of adolescents aged less than 20 years reported a history of self-harm (Hiscock et al., 2018). In addition, the prevalence of NSSI has significantly increased in recent years. It was reported by McManus et al. (2019) that the prevalence of self-reported lifetime NSSI increased from 2.4% in 2000 to 6.4% in 2014, with the largest increase in females aged 16–24 years (from 6.5% to 19.7%). Researchers have also proposed an "iceberg" model of self-harm, indicating that the phenomenon of NSSI is often concealed and that most individuals with NSSI do not seek help from others (McMahon et al., 2014; Geulayov et al., 2018).

AIM: To perform a Meta-analysis to explore the correlation between metabolic syndrome and intraocular pressure (IOP). METHODS: We searched PubMed and Embase in November 2017 for studies discussing the relationship between metabolic syndrome components and IOP in patients. Pearson correlation coefficients, odds ratios and standardized betas were extracted from inclusive studies. Heterogeneity and publication bias were checked. RESULTS: Of 295 articles, 10 met inclusion criteria and provided sufficient data for Meta-analysis. Results showed a significant positive relation between metabolic syndrome and IOP (Z=0.47, 95%CI: 0.15-0.79, P=0.005). The five components [waist circumference, hypertriglyceridemia, high blood pressure, high fasting glucose and low high density lipoprotein (HDL)-cholesterol] of metabolic syndrome all showed positive correlation with IOP except the low HDL-cholesterol which had no statistical significance. The pooled Z was 0.08 (95%CI: 0.04-0.12), 0.16 (95%CI: 0.11-0.21), 0.16 (95%CI: 0.10-0.22), 0.30 (95%CI: 0.20-0.40) and 0.12 (95%CI: 0.08-0.16), respectively. Begg's test and Egger's test showed no evidence of significant publication bias of this Meta-analysis. CONCLUSION: Our findings suggest that metabolic syndrome and its components are significantly associated with IOP, besides the HDL-cholesterol. This association may be used to control IOP by intervening the occurrence of metabolic syndrome.

Background: Infectious keratitis after refractive surgery is uncommon, and mycobacterium abscess has never been observed in corneal infections in China. Purpose: To propose an unusual presentation of two cases about Mycobacterium abscess infection after laser in situ keratomileusis (LASIK). Participants: Both cases 1 and 2 were females who exhibited a corneal ulceration after LASIK in the same hospital. Methods: The first patient in the study was a 28-year-old woman who underwent Epi-LASIK surgery in both eyes. She developed an infectious corneal ulcer in one eye after 50 days post-surgery. She was referred to our out-patient clinic 3 months post-surgery with corneal perforation, and was treated with a therapeutic penetrating keratoplasty. The second patient was a 27-year-old woman who developed infectious keratitis in one eye after 2 months after bilateral LASIK and was referred to us 3 months later. Results: Cultures in both the cases were identified as a Mycobacterium abscess, and case 1 was treated with penetrating keratoplasty eventually, while case 2 recovered after sensitive treatment. Conclusions: Infectious keratitis after refractive surgery is uncommon, and Mycobacterium abscess has never been observed in corneal infections. An accurate diagnosis and identification of the causative agent is very important.

Automatic brain tumor segmentation from multi-modality Magnetic Resonance Images (MRI) using deep learning methods plays an important role in assisting the diagnosis and treatment of brain tumor. However, previous methods mostly ignore the latent relationship among different modalities. In this work, we propose a novel end-to-end Modality-Pairing learning method for brain tumor segmentation. Paralleled branches are designed to exploit different modality features and a series of layer connections are utilized to capture complex relationships and abundant information among modalities. We also use a consistency loss to minimize the prediction variance between two branches. Besides, learning rate warmup strategy is adopted to solve the problem of the training instability and early over-fitting. Lastly, we use average ensemble of multiple models and some post-processing techniques to get final results. Our method is tested on the BraTS 2020 online testing dataset, obtaining promising segmentation performance, with average dice scores of 0.891, 0.842, 0.816 for the whole tumor, tumor core and enhancing tumor, respectively. We won the second place of the BraTS 2020 Challenge for the tumor segmentation task.

BackgroundEndometriosis is a chronic hormonal inflammatory disease characterized by the presence of endometrial tissue (glands and stroma) outside the uterus. Endometriosis seriously affects the physical health of women of childbearing age, often causes infertility, and affects the body and mind of patients and their families.MethodsWe examined the effect of HSF1 on endometriosis through cell count, scratch and clone formation experiments. We used real-time quantitative PCR and western blotting to detect the effect of HSF1 on mRNA and protein of endometriosis cells. Collect the cell culture medium and Glucose levels and lactate levels were determined using a glucose (GO) assay kit and a lactate assay kit. Furthermore, we established a mouse model of endometriosis, and the effect of HSF1 on endometriosis was observed by inhibiting HSF1 with KRIBB11 in the mice.ResultsHSF1 is highly expressed in endometriosis and plays an indispensable role in endometriosis development in both cell and animal studies. We found that HSF1 promotes endometriosis development and glucose consumption and lactic acid production. Further research showed that HSF1 functions in endometriosis by up-regulating PFKFB3, a key enzyme in glycolysis. And the HSF1 inhibitor KRIBB11 can abrogate all of the above experimental effects both in vivo and in vitro. ConclusionsOur study shows that HSF1 plays a significant role in the occurrence and development of endometriosis, which may become a new target for the treatment of endometriosis and provide a new idea for the clinical treatment of endometriosis.

Bacteria-based therapy has shown great promise in treating various diseases by regulating intestinal flora balance in pre-clinical and clinical studies. However, weak resistance against stress in the gastrointestinal (GI) tract and limited retention time in the intestine result in low bacteria availability and persistence, restricting further clinical application. Here, we report a double-layer coating strategy employing tannic acid (TA) and enteric L100 for probiotics encapsulation to address the delivery challenges after oral administration. The Escherichia coli Nissle 1917 (EcN) sequentially encapsulated by TA and L100 layers display robust resistances against the harsh external environment of the GI tract. Furthermore, the pH-responsive degradation of the outer L100 layer leads to the selective delivery of TA-EcN to the intestine, where the strong mucoadhesive capability of the TA layer prolongs the retention time of EcN without compromising the viability and proliferation capabilities of EcN, resulting in superior efficacy of prophylactic and treatment efficacy against colitis. To mitigate the potential side effects caused by long-term mucoadhesion of TA, the TA layer can be further removed by addition of ethylenediaminetetraacetic acid (EDTA). The spatiotemporal delivery of probiotics through double-layer encapsulation can augment the therapeutic efficacy of bacteria-based therapy against GI tract-related diseases.

To design and develop artificial intelligence (AI) hydrocephalus (HYC) imaging diagnostic model using a transfer learning algorithm and evaluate its application in the diagnosis of HYC by non-contrast material-enhanced head computed tomographic (CT) images. A training and validation dataset of non-contrast material-enhanced head CT examinations that comprised of 1000 patients with HYC and 1000 normal people with no HYC accumulating to 28,500 images. Images were pre-processed, and the feature variables were labeled. The feature variables were extracted by the neural network for transfer learning. AI algorithm performance was tested on a separate dataset containing 250 examinations of HYC and 250 of normal. Resident, attending and consultant in the department of radiology were also tested with the test sets, their results were compared with the AI model. Final model performance for HYC showed 93.6% sensitivity (95% confidence interval: 77%, 97%) and 94.4% specificity (95% confidence interval: 79%, 98%), with area under the characteristic curve of 0.93. Accuracy rate of model, resident, attending, and consultant were 94.0%, 93.4%, 95.6%, and 97.0%. AI can effectively identify the characteristics of HYC from CT images of the brain and automatically analyze the images. In the future, AI can provide auxiliary diagnosis of image results and reduce the burden on junior doctors.

Nonalcoholic fatty liver disease (NAFLD) is the hepatic manifestation of metabolic syndrome as an arising health issue globally. Obesity and diabetes seem coherent with high NAFLD occurrence. Currently, its underlying mechanism of cause and progression remains to be elicited and effective therapy is unavailable. Huge investment on studying its pathogenesis and innovative therapy is underway. As mechanistic studies are more difficult or unethical to be performed in humans, animal model that recapitulates the human disease is urgently needed. Nonhuman primates (NHPs) with NAFLD can highly mimic the human disease. This study aimed to induce NAFLD by various diets (Table 1) in normoglycemic and diabetic NHPs with biomarker and histopathology assays to quantify NAFLD occurrence. Our data show that diabetic monkeys with high-fat-high-fructose (HFHF) diet developed NAFLD which could assemble to the human disease and be highly valuable for NAFLD research. However, low choline (LC), LC+10% fructose, LC+low methionine (LCM)+10% fructose diets for 31 weeks had no significant effects on liver function, except significant decrease in body weight and food intake in LCM+10%F group and some increase in liver histological scores in LC+10%F group (Table 1). In summary, normal diet with LC or LCM ± high fructose is hardly to induce NAFLD in normoglycemic NHPs, at least in 31 weeks. HFHF could induce or enhance NAFLD in diabetic NHPs. Disclosure J. Gao: Employee; Self; Crown Bioscience. D. Wang: Employee; Self; Crown Bioscience. Y. Ji: Employee; Self; Crown Bioscience. X. Zhang: Employee; Self; Crown Bioscience. C. Yang: None. J. Zhang: Employee; Self; Crown Bioscience. W. Ye: Employee; Self; Crown Bioscience. F. Shi: Employee; Self; Crown Bioscience. M. Jiang: Employee; Self; Crown Bioscience. Q. Jin: Employee; Self; Crown Bioscience. D. Zhang: None. X. Wang: None. W. Song: Employee; Self; Crown Bioscience. K. Chng: None. H. Jiang: Employee; Self; SYSE Bio-tec Co., Ltd. Y. Wang: None. Y. Xiao: Employee; Self; Crown Bioscience.

MS-NASH mouse is a leptin pathway intact model of spontaneous metabolic diseases. Our previous work developed a nonalcoholic steatohepatitis (NASH) model in MS-NASH mice fed with western diet and fructose drinking water (WDF). This study showed aggravated liver cholesterol content and fibrosis score by using a modified Amylin diet (GAN). Liver Cholesterol was increased from 10.11±1.58 mg/g to 14.07±0.81 mg/g and fibrosis score increased from 1.5±0.3 to 2.1±0.2 when comparing GAN with WDF and with C57Bl/6 mice. Administration of 30 mg/kg Obeticholic Acid (OCA, QD, PO) or 30 mg/kg Elafibranor (GFT-505, QD, PO) for 8 weeks significantly reduced serum liver enzymes, liver lipids and fibrosis scores. Conclusion: The GAN diet fed MS-NASH mice can be a better NASH model for novel compound efficacy. Disclosure G. Zhang: None. K. Chng: None. Y. Wang: None.

Summary: Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was reported from China in January, 2020. SARS-CoV-2 is efficiently transmitted from person to person and, in 2 months, has caused more than 82 000 laboratory-confirmed cases of coronavirus disease 2019 (COVID-19) and 2800 deaths in 46 countries. The total number of cases and deaths has surpassed that of the 2003 severe acute respiratory syndrome coronavirus (SARS-CoV). Although both COVID-19 and severe acute respiratory syndrome (SARS) manifest as pneumonia, COVID-19 is associated with apparently more efficient transmission, fewer cases of diarrhoea, increased mental confusion, and a lower crude fatality rate. However, the underlying virus–host interactive characteristics conferring these observations on transmissibility and clinical manifestations of COVID-19 remain unknown. Methods: We systematically investigated the cellular susceptibility, species tropism, replication kinetics, and cell damage of SARS-CoV-2 and compared findings with those for SARS-CoV. We compared SARS-CoV-2 and SARS-CoV replication in different cell lines with one-way ANOVA. For the area under the curve comparison between SARS-CoV-2 and SARS-CoV replication in Calu3 (pulmonary) and Caco2 (intestinal) cells, we used Student's t test. We analysed cell damage induced by SARS-CoV-2 and SARS-CoV with one-way ANOVA. Findings: SARS-CoV-2 infected and replicated to comparable levels in human Caco2 cells and Calu3 cells over a period of 120 h (p=0·52). By contrast, SARS-CoV infected and replicated more efficiently in Caco2 cells than in Calu3 cells under the same multiplicity of infection (p=0·0098). SARS-CoV-2, but not SARS-CoV, replicated modestly in U251 (neuronal) cells (p=0·036). For animal species cell tropism, both SARS-CoV and SARS-CoV-2 replicated in non-human primate, cat, rabbit, and pig cells. SARS-CoV, but not SARS-CoV-2, infected and replicated in Rhinolophus sinicus bat kidney cells. SARS-CoV-2 consistently induced significantly delayed and milder levels of cell damage than did SARS-CoV in non-human primate cells (VeroE6, p=0·016; FRhK4, p=0·0004). Interpretation: As far as we know, our study presents the first quantitative data for tropism, replication kinetics, and cell damage of SARS-CoV-2. These data provide novel insights into the lower incidence of diarrhoea, decreased disease severity, and reduced mortality in patients with COVID-19, with respect to the pathogenesis and high transmissibility of SARS-CoV-2 compared with SARS-CoV. Funding: May Tam Mak Mei Yin, The Shaw Foundation Hong Kong, Richard Yu and Carol Yu, Michael Seak-Kan Tong, Respiratory Viral Research Foundation, Hui Ming, Hui Hoy and Chow Sin Lan Charity Fund, Chan Yin Chuen Memorial Charitable Foundation, Marina Man-Wai Lee, The Hong Kong Hainan Commercial Association South China Microbiology Research Fund, The Jessie & George Ho Charitable Foundation, Perfect Shape Medical, The Consultancy Service for Enhancing Laboratory Surveillance of Emerging Infectious Diseases and Research Capability on Antimicrobial Resistance for the Department of Health of the Hong Kong Special Administrative Region Government, The Theme-Based Research Scheme of the Research Grants Council, Sanming Project of Medicine in Shenzhen, and The High Level-Hospital Program, Health Commission of Guangdong Province, China.

Background: Trials reported there are beneficial effects of the addition of bevacizumab to chemotherapy in advanced cervical cancer but might have adverse effects. The purposes of the study were to evaluate the treatment response and safety of the addition of bevacizumab to paclitaxel plus carboplatin in advanced cervical cancer women. Methods: Data on treatment response, adverse effects, and overall survival of women who received paclitaxel plus carboplatin every 3 weeks (ACT cohort, n = 161) or paclitaxel, carboplatin, and bevacizumab every 3 weeks (PCB cohort, n = 127) until disease progression or severe adverse events were collected and analyzed. Results: The treatment response of paclitaxel plus carboplatin increased on addition of bevacizumab ( P = .037). Neutropenia (grade ≥3, P = .001), leukopenia (grade 4, P = .041), anemia (grade ≥3, P = .031), hypertension (grade ≥2, P = .002), and gastrointestinal fistula (grade ≥2, P = 0.006) are reported in the PCB cohort. Women of ACT and PCB cohorts reported an overall survival of 20.11 ± 3.15 months and 24.52 ± 4.05 months, respectively. Conclusions: Addition of bevacizumab increases the treatment response of paclitaxel and carboplatin chemotherapy and overall survival of women with advanced cervical cancers, but it is not well tolerated.

ObjectiveWe aimed to investigate clinical features and management of 55 COVID-19 patients in Wuxi, especially severe COVID-19.MethodsEpidemiological, demographic, clinical, laboratory, imaging, treatment, and outcome data of patients were collected. Follow-up lasted until April 6, 2020.ResultsAll 55 patients included 47 (85.5%) non-severe patients and 8 (14.5%) severe patients. Common comorbidities were hypertension and diabetes. Common symptoms were fever, cough and sputum. Lymphopenia was a common laboratory finding, and ground-glass opacity was a common chest CT feature. All patients received antiviral therapy of α-interferon inhalation and lopinavir-ritonavir tablets. Common complications included acute liver injury and respiratory failure. All patients were discharged. No death was occurred and no medical staff got infected. Patients with severe COVID-19 showed significantly older age, decreased lymphocytes, increased C reactive protein, and higher frequency of bilateral lung infiltration compared to non-severe patients. Significantly more treatments including antibiotic therapy and mechanical ventilation, longer hospitalization stay and higher cost were shown on severe patients.ConclusionsOur study suggested that patients with severe COVID-19 may be more likely to have an older age, present with lymphopenia and bilateral lung infiltration, receive multiple treatments and stay longer in hospital.

Avian leukosis (AL) is one of the most pandemic immunosuppressive diseases and has been widely spread between 2006 and 2009 in China. The contamination of avian leukosis virus (ALV) in attenuated vaccine is considered as one of the possible transmission routes of this disease. Based on a retrospective survey of 918 batches of attenuated vaccine produced before 2010, three of them were identified as ALV-positive and corresponding ALV strains were successfully isolated from a live Fowlpox virus vaccine, a live Newcastle disease virus vaccine and a live Infectious Bursal Disease virus vaccine, respectively, and whole-genome sequencing showed that these three isolates shared the highest homology with ALV-A wild strains isolated in China (97.7%) over the same period, and the phylogenetic analysis based on their gp85 genes further confirmed that they belong to subgroup A. Meanwhile, although these three ALV-A strains isolated from contaminated vaccines shared a close genetic relationship, their U3 region of genome have a relatively low identity, suggesting that these three strains may have different sources. This study reminds us once again that the possibility of ALV infecting chickens through contaminated live vaccines, requiring us to carry out stricter exogenous virus monitoring in vaccines.

Corneal endothelial disease is a global sight-threatening disease, and corneal transplantation using donor corneas remains the sole therapeutic option. A previous work demonstrated that N (2)-alanyl-glutamine (Ala-Gln) protected against apoptosis and cellular stress, and maintained intestinal tissue integrity. In this pursuit, the present study aimed to examine the effect of Ala-Gln in the protection of the corneal endothelium and expand its range of potential clinical applications. Mice in the control group were intracamerally irrigated with Ringers lactate injection, whereas those in the experimental group were irrigated with Ringers lactate injection containing Ala-Gln. The mean intraocular pressure increased to 44 ± 3.5 mm Hg during intracameral irrigation (normal range 10.2 ± 0.4 mmHg). In vivo confocal microscopy results showed that the addition of Ala-Gln protected the morphology, structure, and density of the corneal endothelial cells. Optical Coherence Tomography (OCT) measurements showed that corneal thickness was not significantly different between the two groups, because of the immediate corneal edema after irrigation, but the addition of Ala-Gln obviously promoted the recovery of the corneal edema. Scanning electron microscopy indicated that the corneal endothelial cells were severely ruptured and exfoliated in the Ringer's group accompanied with cellular edema, when compared with the Ala-Gln group. The intracameral irrigation using Ala-Gln protected the structure and expression of cytoskeleton and Na-K-ATPase, which exhibited a regular distribution and significantly increased expression in comparison to Ringer's group. Furthermore, Ala-Gln maintained the mitochondrial morphology and increased the activity of mitochondria. Moreover, transmission electron microscopy showed that intracameral irrigation of Ala-Gln reversed the ultrastructural changes induced by the acute ocular hypertension in mice. Our study demonstrates that the intracameral irrigation of Ala-Gln effectively maintained the corneal endothelial pump function and barrier function by protecting the mitochondrial function and preventing the rearrangement of cytoskeleton in acute ocular hypertension in mice.

To better inform the next clinical trials of vascular disrupting agent combretastatin-A4-phosphate (CA4P) in patients with hepatic malignancies, this preclinical study aimed at evaluating CA4P therapeutic efficacy in rats with primary hepatocellular carcinomas (HCCs) of a full spectrum of differentiation and vascularity by magnetic resonance imaging (MRI), microangiography and histopathology. Ninety-six HCCs were raised in 25 rats by diethylnitrosamine gavage. Tumor growth was monitored by T2-/T1-weighted-MRI (T2WI, T1WI) using a 3.0 T scanner. Early vascular response and later intratumoral necrosis were detected by dynamic-contrast-enhanced (DCE) MRI and diffusion-weighted-imaging (DWI) before, 1 and 12 hr after CA4P iv-administration. In vivo MRI-findings were validated by postmortem-techniques. Multi-parametric MRI revealed rapid CA4P-induced tumor vascular shutdown within 1 hr, followed by variable intratumoral necrosis at 12 hr. Tumor volumes decreased by 10% at 1 hr (p

Background Reticuloendotheliosis is an immunosuppressive disease caused by avian reticuloendotheliosis virus (REV). It is commonly found in poultry farms and has caused a notable economic loss worldwide. Despite this, there is currently no effective vaccine available to protect against REV infection. Method In this study, gp90 protein derived from an REV isolated from a contaminated vaccine was co-administered with cytosine-phosphate-guanine oligodeoxynucleotide (CpG-ODN) adjuvant to hens to determine if it protects their chicks against REV infection. To synthesize the gp90 protein, the gp90 gene was amplified using polymerase chain reaction, expressed in Escherichia coli, and purified. The resulting recombinant protein was injected intramuscularly into breeder hens along with CpG-ODN adjuvant and then serum antibody levels were regularly evaluated. After the fertilized eggs from these vaccinated hens had hatched, the resulting chicks were challenged with a 102.7 50% tissue culture infectious dose (TCID50) of REV at 1 day old and the REV antibody levels in these hatched chickens were evaluated before and after the challenge. Viremia and growth rate were measured weekly and statistically analyzed. Results The results suggest that the gp90 recombinant protein was successfully prepared and, when used with CpG-ODN adjuvant to immunize breeder hens, induced serological antibody production against REV in both hens and their hatched chicks. In addition, the maternal antibodies induced by the gp90 protein vaccine effectively protected majority of the chicks from REV infection. Conclusions Overall, we found the gp90 protein obtained in this study may be a potential vaccine candidate that had good immunogenicity and could be an auxiliary measure to accelerate the eradication of REV.

Background Previous studies have shown that serum interleukin 33 serving as an “alarmin” is increased in children with asthma. The objective of this study was to assess the validity of serum IL33 test for early diagnosis of childhood asthma. Methods A literature search was performed in June 2016 using PubMed, Embase, the Cochrane Library and other Chinese Medical Databases to identify studies. The search terms used were “cytokine”, “interleukin-33“, “asthma” and “children”. The meta-analysis was performed using Review Manager 5.3 software. Random-effects model was used to estimate the standardized mean differences (SMDs) with 95% confidence intervals (CIs). Results A total of eight studies were included into this meta-analysis, involving 330 asthmatic children and 248 healthy children. The meta-analysis results revealed that the serum IL33 level was higher in asthmatic children compared to that in healthy children (SMD = 1.29, 95%CI = 0.53–2.05, P = 0.0009), with significant heterogeneity across studies ( I 2 = 94% and P Conclusions The meta-analysis showed that serum IL33 is a helpful biomarker for early diagnosis of childhood asthma. However, owing to lack of enough data, the increased serum concentration of IL33 cannot be an indicator for the asthma severity.

Background Avian leukosis virus (ALV) is one of the main causes of tumour development within the poultry industry in China. The subgroup J avian leukosis viruses (ALV-J), which induce erythroblastosis and myelocytomatosis, have the greatest pathogenicity and transmission ability within this class of viruses. ALV can be transmitted both horizontally and vertically; however, the effects of ALV infection in chickens—especially roosters—during the propagation, on future generations is not clear. Knowing the role of the cock in the transmission of ALV from generation to generation might contribute to the eradication programs for ALV. Results The results showed that two hens inseminated with ALV-J-positive semen developed temporary antibody responses to ALV-J at 4–5 weeks post insemination. The p27 antigen was detected in cloacal swabs of six hens, and in 3 of 26 egg albumens at 1–6 weeks after insemination. Moreover, no viremia was detected at 6 weeks after insemination even when virus isolation had been conducted six times at weekly intervals for each of the 12 females. However, ALV-J was isolated from 1 of their 34 progeny chicks at 1 week of age, and its gp85 had 98.4%–99.2% sequence identity with the gp85 of ALV-J isolated from semen samples of the six cocks. Conclusions Our findings indicated that females that were late horizontally infected with ALV-J by artificial insemination might transmit the virus to progeny through eggs, which amounts to vertical transmission.

The use of antiplatelet therapy after intracerebral hemorrhage remains controversial, while the use of dual antiplatelet therapy (DAPT) is required after cardiac stenting. In this study, we examine the risk of bleeding and ischemic events for PCI patients with a history of hypertensive hemorrhage on DAPT.A total of 128 cases and 153 controls were selected from Chinese patients with cardiac stenting on dual anti-platelet therapy for a single-center retrospective case-control study. Patients with a history of hypertensive hemorrhage were selected for the case group, while patients with a history of hypertension were chosen as control. All patients were on aspirin 100 mg and clopidogrel 75 mg after cardiac stenting, and were followed for a duration of 12-48 months. The primary outcomes were intracerebral hemorrhage, major bleeding, and major adverse cardiovascular and cerebrovascular events.A history of previous hypertensive hemorrhage was not found to be a risk factor for intracerebral hemorrhage and major bleeding while on dual anti-platelet therapy. However, a history of either hypertensive hemorrhage or coronary artery disease was independently found to be risk factors for major adverse cardiovascular and cerebrovascular events. On sub-group analysis, patients with a history of hypertensive hemorrhage within 12 months were found to be at higher risk for bleeding on dual anti-platelet therapy, while patients with history of hypertensive hemorrhage outside of 12 months on dual anti-platelet therapy did not have the same increased risk.A history of hypertensive hemorrhage and coronary heart disease were two independent risk factors for major adverse cardiovascular and cerebrovascular events in PCI patients taking DAPT. A history of hypertensive hemorrhage less than 12 months had an increased risk for recurrent intracerebral hemorrhage and major bleeding in PCI patients taking DAPT.

Insulin resistance and diabetes can develop spontaneously with obesity and aging in rhesus monkeys, highly similar to the natural history of obesity, insulin resistance, and progression to type 2 diabetes in humans. The current studies in obese rhesus were undertaken to assess hepatic and adipose contributions to systemic insulin resistance—currently, a gap in our knowledge—and to benchmark the responses to pioglitazone (PIO). A two-step hyperinsulinemic-euglycemic clamp, with tracer-based glucose flux estimates, was used to measure insulin resistance, and in an intervention study was repeated following 6 wk of PIO treatment (3 mg/kg). Compared with lean healthy rhesus, obese rhesus has a 60% reduction of glucose utilization during a high insulin infusion and markedly impaired suppression of lipolysis, which was evident at both low and high insulin infusion. However, obese dysmetabolic rhesus manifests only mild hepatic insulin resistance. Six-week PIO treatment significantly improved skeletal muscle and adipose insulin resistance (by ~50%). These studies strengthen the concept that insulin resistance in obese rhesus closely resembles human insulin resistance and indicate the value of obese rhesus for appraising new insulin-sensitizing therapeutics.

In our previous study, six subgroup J strains of avian leukosis virus (ALV-J)-associated acutely transforming viruses carrying different lengths of the v-fps oncogene, designated as Fu-J and Fu-J1-5, were isolated and characterized from fibrosarcomas in ALV-J-infected chickens. In the present study, the oncogenic potential of Fu-J and Fu-J1-5 was investigated using a reverse genetics technique. Six replication-defective viruses, named rFu-J and rFu-J1-5, were rescued with the replication-competent rescued ALV-J strain rSDAU1005 as a helper virus by co-transfection of chicken embryo fibroblast monolayers with infectious clone plasmids. Experimental bird studies were performed, demonstrating that only the rescued rFu-J virus carrying the complete v-fps oncogene with rSDAU1005 as the helper virus could induce acute fibrosarcoma after inoculation in specific-pathogen-free (SPF) chickens. These results provide direct evidence that the replication-defective acutely transforming Fu-J virus, with the complete v-fps oncogene, was associated with acute fibrosarcoma in chickens infected with ALV-J in the field, as reported previously.

Acute lung injury (ALI) and idiopathic pulmonary fibrosis (IPF) are severe lung diseases causing irreversible lung damage and premature death. Both diseases share multiple pathological features, including overexpression of C-X-C chemokine receptor type 4 (CXCR4) and upregulation of plasminogen activator inhibitor-1 (PAI-1). The goal of the present study was to evaluate therapeutic potential of pulmonary treatment with combined inhibition of CXCR4 and PAI-1 in ALI and various disease stages of IPF. We report preparation of perfluorocarbon emulsion polyplexes containing a fluorinated polymeric CXCR4 antagonist (F-PAMD) as an siRNA carrier suitable for pulmonary delivery. In vitro testing of the emulsion polyplexes in primary lung fibroblasts from IPF mice showed high cellular uptake and promising antifibrotic effect as indicated by the decreased expression of α smooth muscle actin, when compared with conventional siRNA polyplexes. Biodistribution analysis in mice with IPF showed prolonged lung retention and widespread lung distribution following intratracheal administration of the formulations. The emulsion polyplexes showed promising therapeutic efficacy in ALI and in early fibrinogenic stage of IPF. Increased survival was observed in the model of late-stage IPF. The use of perfluorocarbon emulsion polyplexes to achieve combined CXCR4 antagonism and PAI-1 inhibition is a promising strategy for treatment of ALI and IPF.

Carbon Tetrachloride (CCl4) is a common hepatotoxin used in liver injury and fibrosis animal models, but it is rare to observe obesity and liver fat accumulation in CCl4-induced liver fibrosis models since weight loss is also provoked. Nonalcoholic Steatohepatitis (NASH) is an epidemic disease coincides with obesity and type 2 diabetes (T2D) causing liver steatosis and inflammation. There are no perfect animal models that capture all steatosis, inflammation and following fibrosis stages of NASH disease, our recent study developed an accelerated fibrotic model in previously established NASH model of western diet fed FATZO/pco mice. Using a dose of 0.08 mL/kg (IP, 2/week) CCl4 on 8-week western diet fed FATZO/pco mice, the average histology fibrosis score increased from 1.2±0.2 to 3.0±0.0, average lobular inflammation score increased from 1.4±0.7 to 1.9±0.1, while average steatosis score remained stable. Hepatotoxicity of CCl4 affected the hepatocyte ballooning score due to increased necrosis, reducing ballooning score from 1.6±0.4 to 1.0±0.0. It is also interesting to find out the FATZO/pco strain tolerated CCl4 challenges better than the control C57Bl/6 strain. Conclusion: The study developed an accelerated NASH mice model with balanced performance in all stages of histopathology and provides better translational values in comparison with human patients. Disclosure G. Zhang: None. K. Chng: None. Y. Wang: None.

The FATZO mouse is a polygenic model with an intact leptin system and was developed by cross breeding of C57BL6/J (C57) with AKR/J. FATZO mice have been used in many dysmetabolic studies. The epidemic health issue of nonalcoholic fatty liver disease (NAFLD, including NASH) is the organ dysmetabolic manifestation. This study was to investigate a high fat/fructose diet (HFFD) on liver function in FATZO mice. The body weights of FATZO (n=30) at age of 9 to 10 weeks were significantly heavier than those of age-matched C57 (n=19) mice (39±0.4g vs. 23±0.2g, p Disclosure X. Wang: None. W. Ye: None. Y. Chen: None. X. Zhang: None. Y. Wang: None. Y. Xiao: Employee; Self; Crown Bioscience Inc.

Lung metastasis is a common and deadly occurrence in many types of solid tumors. Chemokine receptor CXCR4 and transcription factor signal transducer and activator of transcription 3 (STAT3) are among potential therapeutic targets in lung metastatic cancer. Both CXCR4 and STAT3 play important roles in the proliferation, angiogenesis, and metastasis of cancer cells. Here, we report on the development of a pulmonary delivery (p.d.) system based on perfluorocarbon (PFC) nanoemulsions for combined delivery of a partially fluorinated polymeric CXCR4 antagonist (FM) and anti-STAT3 small interfering RNA (siRNA). We have prepared FM-stabilized PFC (FM@PFC) as a delivery system of therapeutic siRNA adsorbed on the surface of the emulsion. These FM@PFC/siRNA nanoemulsions inhibited both CXCR4 and STAT3, as demonstrated by effective anti-invasive ability in vitro and related antimetastatic activity in vivo. The combined nanoemulsions provided a comprehensive anticancer effect in the model of established lung metastasis of breast carcinoma, which was dependent on induction of cancer cell apoptosis, anti-angiogenic effect, anti-invasive activity, and overcoming of the immunosuppressive tumor microenvironment. Direct comparison with intravenous (i.v.) injection showed superior activity of pulmonary administration as indicated by significantly increased animal survival. Overall, this work established the suitability of the PFC nanoemulsions for p.d. of combination anticancer treatments and as a promising method to treat lung metastasis.

Osteoarthritis (OA) is a chronic joint disease resulting from joint inflammation and damage. In this study, we employed a boundary lubricant known as a 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) liposome for loading of an anti-inflammatory drug D-glucosamine sulphate (GAS) to construct a treatment strategy allowing for sustained anti-inflammation and reduced damage. This kind of drug-loaded nanocarrier integrates the anti-inflammatory effect of the GAS and the lubrication ability of DSPC liposomes without the involvement of complex synthesis processes leading to easier popularization. Our experimental results indicated that the GAS-loaded DSPC liposomes could release GAS in a sustained manner while providing good lubrication in pure water (H2O) and phosphate buffered saline (PBS). Moreover, the GAS-loaded DSPC liposomes prepared at a 2 : 8 molar ratio in PBS exhibited a greater entrapment efficiency, lower GAS release rate and smaller friction coefficient as compared to those prepared in H2O. The superiority of the drug release and lubrication ability achieved with the GAS-loaded DSPC liposomes in PBS were elucidated on the basis of salt-induced enhancement in liposomal stability and hydration lubrication by the hydrated salt ions. Such GAS release accelerated the viability and proliferation of primary mouse chondrocytes while also providing the anti-inflammatory and chondroprotective potential for tumor necrosis factor (TNF-α) induced chondrocyte degeneration through the down-regulation of pro-inflammatory cytokines, pain related gene and catabolic proteases, as well as the up-regulation of anabolic components. We envision that the GAS-loaded DSPC liposomes could represent a promising new strategy for clinical treatment of OA in the future.

Diabetic retinopathy is a diabetes complication. During diabetic retinopathy development, the TLR4/NF-κB pathway is up-regulated. Apocynin is a nicotinamide adenine dinucleotide phosphate oxidase blocker which can reduce the superoxide radicals. It is demonstrated that apocynin can inhibit TLR4/NF-κB pathway in rats. We aim to figure out whether apocynin treatment is benefit for the diabetic retinopathy in rat model. The diabetes in rats was induced by streptozotocin. The treatment of apocynin (16 mg/kg/day) or vehicle in diabetic rat model was maintained for 12 weeks. The expression levels of relative genes in this research were shown through Western blot and qRT-PCR. Morphology of the retinas was shown by Hematoxylin-Eosin staining. The treatment of apocynin ameliorated biochemical indexes in diabetic rats and rescued the morphology of the retinas. After a 12 weeks apocynin treatment, the cell apoptosis, oxidative stress, and inflammatory in retina was reduced in diabetic rats. TLR4/NF-κB signaling pathway activity in diabetic rat retina was inhibited by apocynin. Based on our study, the treatment of apocynin ameliorates diabetic retinopathy in rats. The TLR4/NF-κB signaling pathway inhibition by apocynin is involved in this process. This result indicated a great therapeutic potential of apocynin in diabetic retinopathy treatment.

Background Metabolic disorders such as insulin resistance, obesity, and hyperglycemia are prominent risk factors for the development of non-alcoholic fatty liver disease (NAFLD)/steatohepatitis (NASH). Dietary rodent models employ high fat, high cholesterol, high fructose, methionine/choline deficient diets or combinations of these to induce NAFLD/NASH. The FATZO mice spontaneously develop the above metabolic disorders and type 2 diabetes (T2D) when fed with a normal chow diet. The aim of the present study was to determine if FATZO mice fed a high fat and fructose diet would exacerbate the progression of NAFLD/NASH. Methods Male FATZO mice at the age of 8 weeks were fed with high fat Western diet (D12079B) supplemented with 5% fructose in the drinking water (WDF) for the duration of 20 weeks. The body weight, whole body fat content, serum lipid profiles and liver function markers were examined monthly along with the assessment of liver histology for the development of NASH. In addition, the effects of obeticholic acid (OCA, 30 mg/kg, QD) on improvement of NASH progression in the model were evaluated. Results Compared to normal control diet (CD), FATZO mice fed with WDF were heavier with higher body fat measured by qNMR, hypercholesterolemia and had progressive elevations in AST (~ 6 fold), ALT (~ 6 fold), liver over body weight (~ 2 fold) and liver triglyceride (TG) content (1.4–2.9 fold). Histological examination displayed evidence of NAFLD/NASH, including hepatic steatosis, lobular inflammation, ballooning and fibrosis in FATZO mice fed WDF. Treatment with OCA for 15 weeks in FATZO mice on WDF significantly alleviated hypercholesterolemia and elevation of AST/ALT, reduced liver weight and liver TG contents, attenuated hepatic ballooning, but did not affect body weight and blood TG levels. Conclusion WDF fed FATZO mice represent a new model for the study of progressive NAFLD/NASH with concurrent metabolic dysregulation.

The case characteristics of lung cancer are extremely complex, difficult to distinguish, and the rate of deterioration is rapid, and its early symptoms are not obvious. Early diagnosis and treatment of lung cancer is one of the main directions to reduce lung cancer mortality. The computer-aided detection and diagnosis system reduces the workload of the physician and improves the accuracy of image reading. In this paper, based on the analysis of the current lung nodule segmentation algorithm, in order to enhance the accuracy of lung nodule segmentation extraction, the adaptive particle swarm optimization algorithm is used to realize the simultaneous optimization of the number of mixed components and the model parameters, and finally realize the segmentation of lung nodules. The effectiveness and accuracy of segmentation of lung nodule recognition by adaptive particle swarm optimization algorithm is verified by adaptive particle swarm optimization and image model establishment. Provide new aids for the identification of pulmonary nodules.

Bisphenol A (BPA), one of the most prevalent chemicals for daily use, was recently reported to disturb the homeostasis of energy metabolism and insulin signaling pathways, which might contribute to the increasing prevalence rate of mild cognitive impairment (MCI). However, the underlying mechanisms are remained poorly understood. Here we studied the effects of low dose BPA on glucose transport and the IR/IRS/AKT/GSK3β axis in adult male mice to delineate the association between insulin signaling disruption and neurotoxicity mediated by BPA. Mice were treated with subcutaneous injection of 100μg/kg/d BPA or vehicle for 30 days, then the insulin signaling and glucose transporters in the hippocampus and prefrontal cortex were detected by western blot. Our results showed that mice treated with BPA displayed significant decrease of insulin sensitivity, and in glucose transporter 1, 3 (GLUT1, 3) protein levels in mouse brain. Meanwhile, hyperactivation of IR/IRS/AKT/GSK3β axis was detected in the brain of BPA treated mice. Noteworthily, significant increases of phosphorylated tau and β-APP were observed in BPA treated mice. These results strongly suggest that BPA exposure significantly disrupts brain insulin signaling and might be considered as a potential risk factor for neurodegenerative diseases.

To understand the cytogenetic characteristics of acute fibrosarcoma in chickens infected with the subgroup J avian leukosis virus associated with the v-src oncogene, we performed a karyotype analysis of fibrosarcoma cell cultures. Twenty-nine of 50 qualified cell culture spreads demonstrated polyploidy of some macrochromosomes, 21 of which were trisomic for chromosome 7, and others were trisomic for chromosomes 3, 4, 5 (sex chromosome w), and 10. In addition, one of them was trisomic for both chromosome 7 and the sex chromosome 5 (w). In contrast, no aneuploidy was found for 10 macrochromosomes of 12 spreads of normal chicken embryo fibroblast cells, although aneuploidy for some microchromosomes was demonstrated in five of the 12 spreads. The cytogenetic mosaicism or polymorphism of the aneuploidy in the acute fibrosarcoma described in this study suggests that the analysed cells are polyclonal.

Methamphetamine (METH), an illicit drug, is widely abused in many parts of the world. Mounting evidence shows that METH exposure contributes to neurotoxicity, particularly for the monoaminergic neurons. However, to date, only a few studies have tried to unravel the mechanisms involved in METH-induced non-monoaminergic neural damage. Therefore, in the present study, we tried to explore the mechanisms for METH-induced neural damage in cortical neurons. Our results showed that METH significantly increased intracellular [Ca(2) (+) ]i in Ca(2) (+) -containing solution rather than Ca(2) (+) -free solution. Moreover, METH also upregulated calmodulin (CaM) expression and activated CaM-dependent protein kinase II (CaMKII). Significantly, METH-induced neural damage can be partially retarded by CaM antagonist W7 as well as CaMKII blocker KN93. In addition, L-type Ca(2) (+) channel was also proved to be involved in METH-induced cell damage, as nifedipine, the L-type Ca(2) (+) channel-specific inhibitor, markedly attenuated METH-induced neural damage. Collectively, our results suggest that Ca(2) (+) -CaM-CaMKII is involved in METH-mediated neurotoxicity, and it might suggest a potential target for the development of therapeutic strategies for METH abuse. Copyright © 2016 John Wiley & Sons, Ltd.

Introduction Breast cancer radiotherapy cures many women, but, as with other therapies, can cause late side-effects. Methods We undertook meta-analyses of individual patient data from the trials of breast cancer radiotherapy, relating various characteristics of the regimens tested to cause-specific mortality rate ratios (RRs) and second cancer incidence RRs. Doses to cardiac structures were calculated for trials with some heart disease death(s), lung doses were calculated for megavoltage trials with some lung cancer(s) in the second decade after radiotherapy, and oesophagus doses were calculated for megavoltage trials with some oesophageal cancer(s). Trial radiotherapy regimens were reconstructed for a woman with typical anatomy using virtual simulation and 3-dimensional CT planning (and, for a few regimens, manual planning). Results Information was available on 40,781 women in 75 evenly randomised comparisons of radiotherapy versus not. Median follow-up was 9.7 years and 20,345 died, 6064 without recurrence. Smoking information for included women was unavailable. Mean normal tissue radiation doses for irradiated women were: heart 6.3 Gy (range Allocation to radiotherapy increased non-breast-cancer mortality (RR=1.15, 95% CI 1.09–1.22, 2p Second cancer incidence was increased (RR=1.23, 1.12–1.36, 2p Conclusions Since these trials, normal tissue doses from breast cancer radiotherapy have at least halved so the excess relative risks will be at least halved. Background disease rates have also changed, so the absolute risks will be different for women today. Modelling the effects of such changes suggests that for women who have smoked throughout adult life and will continue smoking, even modern radiotherapy may cause an absolute lung cancer risk of a few per cent, making this the main late side-effect in smokers. However, for non-smokers (and ex-smokers) with healthy hearts who would, under current guidelines, be offered radiotherapy, the expected reduction in breast cancer mortality greatly outweighs any increase in other mortality. Citation Format: Taylor C, Correa C, Anderson S, Duane F, Ewertz M, Jagsi R, Pierce L, Swain S, Whelan T, Wang Z, Wang Y, Peto R, McGale P. Late side-effects of breast cancer radiotherapy: Second cancer incidence and non-breast-cancer mortality among 40,000 women in 75 trials. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr S5-08.

Acute pulmonary edema is one of the major outcomes of exposure to high levels of hydrogen sulfide (H2S). However, the mechanisms involved in H2S-induced acute pulmonary edema are still poorly understood. Therefore, the present study is designed to evaluate the role of epithelial sodium channel (ENaC) in H2S-induced acute pulmonary edema. The Sprague-Dawley rats were exposed to sublethal concentrations of inhaled H2S, then the pulmonary histological and lung epithelial cell injury were evaluated by hematoxylin-eosin staining and electron microscopy, respectively. In addition to morphological investigation, our results also revealed that H2S exposure significantly decreased the alveolar fluid clearance and increased the lung tissue wet-dry ratio. These changes were demonstrated to be associated with decreased ENaC expression. Furthermore, the extracellular-regulated protein kinases 1/2 pathway was demonstrated to be implicated in H2S-mediated ENaC expression, because PD98059, an ERK1/2 antagonist, significantly mitigated H2S-mediated ENaC down-regulation. Therefore, our results show that ENaC might represent a novel pharmacological target for the treatment of acute pulmonary edema induced by H2S and other hazardous gases.

Metastatic breast cancer is a major cause of cancer-related female mortality worldwide. The signal transducer and activator of transcription 3 (STAT3) and the chemokine receptor CXCR4 are involved in the metastatic spread of breast cancer. The goal of this study was to develop nanomedicine treatment based on combined inhibition of STAT3 and CXCR4. We synthesized a library of CXCR4-inhibiting polymers with a combination of beneficial features that included PEGylation, fluorination, and bioreducibility to achieve systemic delivery of siRNA to silence STAT3 expression in the tumors. An in vivo structure-activity relationship study in an experimental lung metastasis model revealed superior antimetastatic activity of bioreducible fluorinated polyplexes when compared with nonreducible controls despite similar CXCR4 antagonism and the ability to inhibit in vitro cancer cell invasion. When compared with nonreducible and nonfluorinated polyplexes, improved siRNA delivery was observed with the bioreducible fluorinated polyplexes. The improvement was ascribed to a combination of enhanced physical stability, decreased serum destabilization, and improved intracellular trafficking. Pharmacokinetic analysis showed that fluorination decreased the rate of renal clearance of the polyplexes and contributed to enhanced accumulation in the tumors. Therapeutic efficacy of the polyplexes with STAT3 siRNA was assessed in early stage breast cancer and late-stage metastatic breast cancer with primary tumor resection. Strong inhibition of the primary tumor growth and pronounced antimetastatic effects were observed in both models of metastatic breast cancer. Mechanistic studies revealed multifaceted mechanism of action of the combined STAT3 and CXCR4 inhibition by the developed polyplexes relying both on local and systemic effects.

Introduction: Cardiomyopathy is the leading cause of morbidity and mortality among all complications of type 2 diabetic (T2D) and obese patients. Diabetic cardiomyopathy (DC) is characterized by an initial cardiac hypertrophy followed by thinning of the cardiac walls with declines in both systolic and diastolic functions, which ultimately leads to heart failure. No rodent models fully captured phenotypes of DC. The ZDSD rat, a new generation of T2D rat model with intact leptin signaling features with slow onset of diabetes and obesity, which closely mimics the development of the disease in patients. Here we sought to evaluate the cardiac function during the development of metabolic syndromes in ZDSD rats. Methods: 12 male ZDSD rats and age-matched SD controls were monitored for blood pressure, glucose, and cardiac function using echocardiography. Animals were also challenged with 1 mg/kg dobutamine for the assessment of cardiac reserve. Results: ZDSD rats developed hypertension from age of 18 weeks. Their left ventricular (LV) functions were compromised along with changes in cardiac morphology. At resting state, ZDSD rats showed LV hypertrophy from age of 18 to 22 weeks after which cardiac walls became thinner with larger LV volume. At 34 weeks old, both ejection fraction (EF) and transmitral E/A ratio declined. Upon treatment with dobutamine for 5 minutes, SD rats reached almost 98% EF, while the values of ZDSD were 91% at 30 and 34 weeks old, suggesting the loss of LV contractility the animals. Conclusion: ZDSD rats which carry multiple dysmetabolic phenotypes are spontaneously hypertensive with reduction in LV function and cardiac reserve which resembles ultrasonic symptoms of diabetic cardiomyopathy patients. Therefore, ZDSD rats may serve as a suitable preclinical model to study potential therapeutic approaches to treat cardiomyopathy with presence of metabolic syndromes. Disclosure G. Sun: None. G. Zhang: None. Y. Wang: None.

Introduction: Type 2 diabetes (T2D) and obesity are strongly associated with peripheral immune cell responses. Subsets of T cells in the circulation are correlated with the severity of the diseases, which may serve as novel targets for antidiabetic or obese treatment. Higher numbers of blood Th1, Th17 and Th22 cells of T helper cells were found in disease patients, while Th2 and Treg cells were in reverse correlation of the severity of the disease. FATZO mouse is a new generation of T2D mouse model with intact leptin signaling. The model is featured with slow onset of diabetes and obesity. Here, we aim to characterize the peripheral immune cell responses in FATZO mice for the use of preclinical drug development targeting diabetes or obesity related inflammation. Methods: Freshly collected blood from 12 FATZO mice at age 6, 12, 22 and 34 weeks were analyzed for the presence of monocytes, B, T, NK, NK-T, CD4+, CD8+ T cells and subsets of T helper cells (Th1, Th2, Th17, Th22, Treg) using FACS. Serum inflammatory cytokines such as IL1, IL2, IL6, IL17, TNFα and IFNγ were also measured. Results: FATZO mice developed obesity and hyperglycemia from age of 12 weeks. Concomitantly, higher numbers of leukocytes were noticed with an increase and decrease in frequency of B and NK cells respectively. Within T cells, numbers of CD4+ T cells decreased by 20%, while CD8+ T cells increased by 25%. When subsets of T helper cells were assessed, percentages of Th1 and Th17 cells significantly elevated during the development of diabetes and obesity, while frequencies of Th2 and Treg cells declined. Serum inflammatory cytokine measurements showed a tendency in higher levels of serum IL1a when the animals became diabetic. Conclusion: FATZO mice develop diabetes and obesity with changes in blood immune cell responses that closely mimic disease patients. The results suggest that FATZO might serve as an ideal model for preclinical research for the diabetes and obesity related inflammation. Disclosure G. Sun: None. W. Ye: Employee; Spouse/Partner; Crown Bioscience Inc.. X. An: None. Y. Xiao: None. Y. Wang: None.

Introduction: Progressive albuminuria and bi-phasic changes in glomular filtration rate (GFR) are hall marks of functional deterioration of the kidney in diabetic nephropathy. However, not all diabetic rodent models can capture the development of such changes in the kidney. FATZO, a new generation of diabetic mouse model with intact leptin signaling pathway shows all the metabolic dysfunctions that mimic human patients such as obesity, dyslipidemia, diabetes and NASH etc. Here, we sought to further characterize the model for the evaluation of its development in diabetic nephropathy. Method: Male FATZO mice on 50diet or on western diet with addition of 10% fructose (WD+fru) from age of 11 weeks were subject to monthly urine collection for albumin and creatinine measurement for 28 weeks, after which animals were treated with Lisinopril (15 mg/kg) and pioglitazone (20 mg/kg) daily for 6 weeks (n=8 for each group). Animals were also monitored for GFR using a subcutaneous monitoring device (Medibeacon, US). Results: Animals on WD+fru were heavier than those on 50diet. Notably, WD+fru markedly elevated urine albumin levels compared to 50diet in FATZO mice which resulted in higher albumin/creatinine ratio and urine albumin excretion rate (UAE) from 2 weeks on diet, while lisinopril treatment for 7 weeks could reduce the levels respectively. In addition, GFR was slightly elevated after 6 weeks of diet induction, which could be normalized by either pioglitazone or lisinopril treatment. Conclusion: We have created a mouse model with progressive albuminuria and elevated GFR that can be reversed by angiotensin-converting-enzyme (ACE) inhibitor. The model with associated dysmetabolic phenotypes including obesity and dyslipidemia can be used as an ideal tool for studying antidiabetic nephropathy treatment. Disclosure G. Sun: None. G. Zhang: None. Y. Wang: None.

Pancreatic β-cell regeneration is considered as a possible therapy for cure of type 2 diabetes (T2D). It has been reported that administration of FTY720, a sphingosine 1-phosphate (S1P) receptor modulator, to db/db mice led to sustained normalization of hyperglycemia by stimulating β-cell in vivo regeneration through PI3K-dependent regulation of cyclin D3 and p57KIP2. To further evaluate the therapeutic potential of FTY720 in the treatment of T2D, we examined the effects of FTY720 on glucose homeostasis in spontaneous diabetic NHP model. NHPs with mean fasting glucose (FG) level of 249 mg/dL were randomly divided into the control (vehicle) and the FTY720-treatment group. FTY720 was orally administered at 5 mg/kg to diabetic NHPs once per day and the FG and HbA1c levels were measured biweekly. Here we show that FTY720 treatment of diabetic NHPs significantly lowered their FG and HbA1c in two weeks. After 10 weeks of treatment, their FG and HbA1c levels remained low and their glucose tolerance was significantly improved. To determine the effect of FTY720 on β-cell function, IVGTT was performed. Following glucose injection, the insulin levels in the control NHPs barely changed. In contrast, the FTY720-treated NHPs secreted large amount of insulin in response to glucose stimulation, consistent with β-cell in vivo regeneration observed in mice. We also evaluated the cardiac function in these NHPs by echocardiography, from which ejection fraction (EF) and fractional shortening (FS) are found abnormally lower in diabetic NHPs (EF=60% and FS=30%) than that in normal NHPs (EF=68% and FS=43%). After 10 weeks of treatment, however, EF in the treatment group was significantly increased from 60% to 70% and FS from 30% to 38% while these values remained abnormally low in the control group. Our data strongly suggest that FTY720 has a great therapeutic promise for the treatment of T2D with benefit to improve the cardiac function in the patients. Disclosure Y. Wang: None. Y. Liu: None. Q. Wei: Employee; Self; Crownbio.Inc.. X. Wang: None. G. Sun: None. Y. Xiao: None. G. Yu: None. Z.A. Ma: None.

1. Abstract 1.1. Introduction: Diabetic Cardiomyopathy (DC) is the leading cause of morbidity and mortality among all complications of Type 2 Diabetes (T2D) and obese patients. It is characterized by an initial cardiac hypertrophy followed by declines in cardiac functions, which ultimately leads to heart failure. No rodent models fully captured phenotypes of DC. The Zucker Diabetic Sprague Dawley (ZDSD) rat, a new generation of T2D rat model with intact leptin signaling features with slow onset of diabetes, obesity and dyslipidemia, which closely mimics the development of the disease in patients. Here we sought to evaluate the cardiac function during the development of metabolic syndromes in ZDSD rats. 1.2. Methods: Age matched male ZDSD and SD rats were monitored for blood pressure and cardiac function using Echocardiography. Animals were also challenged with 1 mg/kg dobutamine for the assessment of cardiac reserve. 1.3. Results: ZDSD rats developed hypertension from age of 18 weeks with both systolic and diastolic blood pressure significantly higher than controls. Their Left Ventricular (LV) functions were compromised along with changes in cardiac morphology. At resting state, ZDSD rats showed thickening of LV posterior wall from age of 18 to 22 weeks after which cardiac walls became thinner with larger LV cavity volume. Both Ejection Fraction (EF) and transmitral E/A ratio of LV declined at 34 weeks old. Upon treatment with dobutamine, ZDSD rats had much lower EF and Fractional Shorterning (FS) compared to SD rats, suggesting the loss of contractility and cardiac reserve of the animals. 1.4. Conclusion: ZDSD rats which carry multiple dysmetabolic phenotypes are spontaneously hypertensive with reduction in LV function and cardiac reserve which resembles ultrasonic symptoms of diabetic cardiomyopathy patients. Therefore, ZDSD rats may serve as a suitable preclinical model to study potential therapeutic approaches to treat cardiomyopathy with presence of metabolic syndromes. 2. Keywords: Cardiomyopathy; Diabetes; Echocardiography; Obesity; Ultrasound

China is going through major change and the incidence of first-ever stroke has increased dramatically. In this study, we aim to determine the ultrasound characteristics of carotid intima-media thickness (CIMT) and carotid plaques (CP) in the Chinese community-based population with high risk of stroke. 1009 stroke-free participants from Datun community were classified at high risk of stroke and included in this cross-sectional study. We performed B-mode carotid ultrasound imaging in all of the study subjects to measure the CIMT in the common carotid artery (CCA) far wall and CP in the CCA, bifurcation and internal carotid artery. Stepwise logistic regression analyses were used to determine factors associated with elevated CIMT and subclinical atherosclerosis, as well as plaque burden (≥2 plaques). Our results showed that traditional risk factors including aging, hypertension, current smoking and the level of high density lipoprotein cholesterol are associated with subclinical atherosclerosis and plaque burden in high-risk community residents. To improve primary prevention in this population, we may consider intense blood pressure and lipid management, and smoking cessation.

Many of the techniques that are employed today by pathologists and oncologists to generate a diagnosis, prognosis or prediction of response have not changed over several decades. However, new molecular and cellular technologies will enable more precise and objective decision-making. This review will detail some of the more recent developments in these areas from Veridex, LLC, academic laboratories and other commercial entities. The discussion of molecular technologies will focus on breast sentinel lymph node biopsy, prostate biopsy, carcinoma of unknown primary, prediction of recurrence in lymph node negative colon and breast cancers and pharmacogenomics. The discussion of cellular technologies will focus on the use of circulating cells to serve in both prognostic and predictive capacities and on the use of molecular methods to interrogate the DNA and RNA isolated from these circulating cells.