Characterisation and Expression Analysis of LdSERK1, a Somatic Embryogenesis Gene in Lilium davidii var. unicolor.

The Lanzhou lily (Lilium davidii var. unicolor) is a variant of the Sichuan lily of the lily family and is a unique Chinese 'medicinal and food' sweet lily. Somatic cell embryogenesis of Lilium has played an important role in providing technical support for germplasm conservation, bulb propagation and improvement of genetic traits. Somatic embryogenesis receptor-like kinases (SERKs) are widely distributed in plants and have been shown to play multiple roles in plant life, including growth and development, somatic embryogenesis and hormone induction. Integrating the results of KEGG enrichment, GO annotation and gene expression analysis, a lily LdSERK1 gene was cloned. The full-length open reading frame of LdSERK1 was 1875 bp, encoding 624 amino acids. The results of the phylogenetic tree analysis showed that LdSERK1 was highly similar to rice, maize and other plant SERKs. The results of the subcellular localisation in the onion epidermis suggested that the LdSERK1 protein was localised at the cell membrane. Secondly, we established the virus-induced gene-silencing (VIGS) system in lily scales, and the results of LdSERK1 silencing by Tobacco rattle virus (TRV) showed that, with the down-regulation of LdSERK1 expression, the occurrence of somatic embryogenesis and callus tissue induction in scales was significantly reduced. Finally, molecular assays from overexpression of the LdSERK1 gene in Arabidopsis showed that LdSERK1 expression was significantly enhanced in the three transgenic lines compared to the wild type, and that the probability of inducing callus tissue in seed was significantly higher than that of the wild type at a concentration of 2 mg/L 2,4-D, which was manifested by an increase in the granularity of the callus tissue.

Interferons are key cytokines acting on pancreatic islets in type 1 diabetes.

AIMS/HYPOTHESIS: The proinflammatory cytokines IFN-α, IFN-γ, IL-1ß and TNF-α may contribute to innate and adaptive immune responses during insulitis in type 1 diabetes and therefore represent attractive therapeutic targets to protect beta cells. However, the specific role of each of these cytokines individually on pancreatic beta cells remains unknown. METHODS: We used deep RNA-seq analysis, followed by extensive confirmation experiments based on reverse transcription-quantitative PCR (RT-qPCR), western blot, histology and use of siRNAs, to characterise the response of human pancreatic beta cells to each cytokine individually and compared the signatures obtained with those present in islets of individuals affected by type 1 diabetes. RESULTS: IFN-α and IFN-γ had a greater impact on the beta cell transcriptome when compared with IL-1ß and TNF-α. The IFN-induced gene signatures have a strong correlation with those observed in beta cells from individuals with type 1 diabetes, and the level of expression of specific IFN-stimulated genes is positively correlated with proteins present in islets of these individuals, regulating beta cell responses to 'danger signals' such as viral infections. Zinc finger NFX1-type containing 1 (ZNFX1), a double-stranded RNA sensor, was identified as highly induced by IFNs and shown to play a key role in the antiviral response in beta cells. CONCLUSIONS/INTERPRETATION: These data suggest that IFN-α and IFN-γ are key cytokines at the islet level in human type 1 diabetes, contributing to the triggering and amplification of autoimmunity.

CD4+ T Cells From Individuals With Type 1 Diabetes Respond to a Novel Class of Deamidated Peptides Formed in Pancreatic Islets.

The ß-cell plays a crucial role in the pathogenesis of type 1 diabetes, in part through the posttranslational modification of self-proteins by biochemical processes such as deamidation. These neoantigens are potential triggers for breaking immune tolerance. We report the detection by LC-MS/MS of 16 novel Gln and 27 novel Asn deamidations in 14 disease-related proteins within inflammatory cytokine-stressed human islets of Langerhans. T-cell clones responsive against one Gln- and three Asn-deamidated peptides could be isolated from peripheral blood of individuals with type 1 diabetes. Ex vivo HLA class II tetramer staining detected higher T-cell frequencies in individuals with the disease compared with control individuals. Furthermore, there was a positive correlation between the frequencies of T cells specific for deamidated peptides, insulin antibody levels at diagnosis, and duration of disease. These results highlight that stressed human islets are prone to enzymatic and biochemical deamidation and suggest that both Gln- and Asn-deamidated peptides can promote the activation and expansion of autoreactive CD4+ T cells. These findings add to the growing evidence that posttranslational modifications undermine tolerance and may open the road for the development of new diagnostic and therapeutic applications for individuals living with type 1 diabetes.

Protection of ß cells against pro-inflammatory cytokine stress by the GDF15-ERBB2 signaling.

Aim/hypothesis: Growth/differentiation factor 15 (GDF15) is a therapeutic target for a variety of metabolic diseases, including type 1 diabetes (T1D). However, the nausea caused by GDF15 is a challenging point for therapeutic development. In addition, it is unknown why the endogenous GDF15 fails to protect from T1D development. Here, we investigate the GDF15 signaling in pancreatic islets towards opening possibilities for therapeutic targeting in ß cells and to understand why this protection fails to occur naturally. Methods: GDF15 signaling in islets was determined by proximity-ligation assay, untargeted proteomics, pathway analysis, and treatment of cells with specific inhibitors. To determine if GDF15 levels would increase prior to disease onset, plasma levels of GDF15 were measured in a longitudinal prospective study of children during T1D development (n=132 cases vs. n=40 controls) and in children with islet autoimmunity but normoglycemia (n=47 cases vs. n=40 controls) using targeted mass spectrometry. We also investigated the regulation of GDF15 production in islets by fluorescence microscopy and western blot analysis. Results: The proximity-ligation assay identified ERBB2 as the GDF15 receptor in islets, which was confirmed using its specific antagonist, tucatinib. The untargeted proteomics analysis and caspase assay showed that ERBB2 activation by GDF15 reduces ß cell apoptosis by downregulating caspase 8. In plasma, GDF15 levels were higher (p=0.0024) during T1D development compared to controls, but not in islet autoimmunity with normoglycemia. However, in the pancreatic islets GDF15 was depleted via sequestration of its mRNA into stress granules, resulting in translation halting. Conclusions/interpretation: GDF15 protects against T1D via ERBB2-mediated decrease of caspase 8 expression in pancreatic islets. Circulating levels of GDF15 increases pre-T1D onset, which is insufficient to promote protection due to its localized depletion in the islets. These findings open opportunities for targeting GDF15 downstream signaling for pancreatic ß cell protection in T1D and help to explain the lack of natural protection by the endogenous protein.

The effectiveness and safety of laparoscopic sleeve gastrectomy with different sizes of bougie calibration: A systematic review and meta-analysis.

OBJECTIVES: This systematic review and meta-analysis was performed to compare the influence of different calibrating bougie sizes on clinical outcomes in laparoscopic sleeve gastrectomy (LSG) for patients with obesity. MATERIALS AND METHODS: A systematic review of the literature was performed using the key words: "laparoscopic sleeve gastrectomy", "bougie size", "calibration", "obesity", and "obese" for searches of electronic databases up to October 2017. Clinical characteristics such as, the percentage of excess weight loss (%EWL), overall complications, gastrointestinal leaks, gastroesophageal reflux disease (GERD) were pooled by meta-analysis. Stata 12.0 (Stata Corp, College Station, TX, USA) was used to perform the meta-analysis. RESULTS: Data were extracted from 11 original studies matching our inclusion criteria. In our review, the group of patients who had operations with thinner bougies had a greater %EWL (SMD 0.23, 95% CI 0.14-0.33, P < .001) than the group where larger diameters were used. Furthermore, no significant differences were found in the incidence of overall complications (OR 1.00, 95% CI 0.73-1.37, P = .978), postoperative gastrointestinal leaks (OR 0.91, 95% CI 0.67-1.24, P = .554), and GERD (OR 0.77, 95% CI 0.37-1.59, P = .476) between the two groups. A robust result could not be made about remission of comorbidities using differing diameter bougies due to insufficient data. CONCLUSIONS: Use of thinner diameter bougies in LSG was more effective in enabling weight loss and did not increase the risk of overall complications, gastrointestinal leaks or GERD compared with larger diameter bougies.

The Effectiveness and Safety of Sleeve Gastrectomy in the Obese Elderly Patients: a Systematic Review and Meta-Analysis.

BACKGROUND: This systematic review was performed to compare the effectiveness and safety of SG in the obese elderly patients with the young ones. METHODS: Cohort studies that compared outcomes among old and young patients who had undergone SG were included. Summary odds ratios were estimated using a random effect model. RESULTS: Eleven studies were included. Old patients had a worse outcome in percentage of excess weight loss than the young ones (SMD -0.39, 95 % CI -0.55 to -0.24). No significant differences were recorded in resolution of co-morbidities: type 2 diabetes mellitus (OR 1.60, 95 % CI 0.84-3.05), hypertension (OR 1.05, 95 % CI 0.65-1.68), dyslipidemia (OR 1.38, 95 % CI 0.68-2.80), OSAS (OR 0.64, 95 % CI 0.30-1.34), or postoperative complications (OR 0.89, 95 % CI 0.51-1.55) between the elderly and the young who had undergone SG. CONCLUSIONS: SG was effective for weight loss and resolution of co-morbidities with low complication rate in the obese elderly patients.

A meta-analysis of maternal and fetal outcomes of pregnancy after bariatric surgery.

BACKGROUND: The effects of bariatric surgery (BS) on outcomes in subsequent pregnancies are unclear. OBJECTIVES: To compare maternal and fetal outcomes among women who become pregnant after BS and obese women who have not undergone BS before pregnancy. SEARCH STRATEGY: PubMed and Embase were searched for relevant reports, and the reference lists of identified articles were hand-searched. SELECTION CRITERIA: Cohort studies that compared outcomes among women who had undergone any type of BS and obese women who had not undergone surgery were included when results were reported as risk ratios or odds ratios (ORs). DATA COLLECTION AND ANALYSIS: Summary ORs were estimated using a random effects model. MAIN RESULTS: Eleven studies were included. Compared with obese women who had not undergone BS, women who had undergone BS had significantly lower odds of gestational diabetes (OR 0.31; 95% CI 0.15-0.65), hypertensive disorders (OR 0.42; 95% CI 0.23-0.78), and macrosomia (OR 0.40; 95% CI 0.24-0.67). However, their odds of small-for-gestational-age newborns were increased (OR 2.16; 95% CI 1.28-3.66). No significant differences were recorded for cesarean, postpartum hemorrhage, and preterm delivery. CONCLUSIONS: BS reduces the odds of some adverse maternal and fetal outcomes among obese women.

MicroRNA-92a as a potential biomarker in diagnosis of colorectal cancer: a systematic review and meta-analysis.

INTRODUCTION: Previous studies demonstrated that MicroRNA-92a (miR-92a) was significantly differential expressed between colorectal cancer (CRC) patients and control cohorts, which provide timely relevant evidence for miR-92a as a novel promising biomarker in the colorectal cancer patients. This meta-analysis aimed to evaluate potential diagnostic value of plasma miR-92a. METHODS: Relevant literatures were collected in PubMed, Embase, Chinese Biomedical Literature Database (CBM), Chinese National Knowledge Infrastructure (CNKI) and Technology of Chongqing (VIP), and Wan Fang Data. Sensitivity, specificity and diagnostic odds ratio (DOR) for miR-92a in the diagnosis of CRC were pooled using random effects models. Summary receiver operating characteristic (SROC) curve analysis and the area under the curve (AUC) were used to estimate the overall test performance. RESULTS: This Meta-analysis included six studies with a total of 521 CRC patients and 379 healthy controls. For miR-92a, the pooled sensitivity, specificity and DOR to predict CRC patients were 76% (95% confidence interval [CI]: 72%-79%), 64% (95% confidence interval [CI]: 59%-69%) and 8.05 (95% CI: 3.50-18.56), respectively. In addition, the AUC of miR-92a in diagnosis CRC is 0.7720. CONCLUSIONS: MicroRNA-92a might be a novel potential biomarker in the diagnosis of colorectal cancer, and more studies are needed to highlight the theoretical strengths.

Alpha-fetoprotein-L3 in hepatocellular carcinoma: a meta-analysis.

BACKGROUND AND OBJECTIVE: Alpha-fetoprotein (AFP) has been widely used as a diagnostic marker. AFP is also increased in patients at high risk for hepatocellular carcinoma (HCC), ie those with chronic hepatitis. The percentage of lens culinaris agglutinin-reactive alpha-fetoprotein (AFP-L3%) has long been proposed as a marker for HCC, but has not been widely adopted due to inconsistent results reported in the literature. In this study, the performance of AFP-L3% is specifically evaluated for diagnosis of HCC. METHODS: A systematic review of relevant studies, the sensitivity, specificity, and diagnostic odds ratio (DOR) for the diagnosis of HCC were pooled using random-effects models. The overall test performance was summarized using summary receiver operating characteristic (SROC) curve analysis. Potential between-study heterogeneity was explored by meta-regression model. RESULTS: Twelve articles were included in this meta-analysis. The overall estimates of AFP-L3% in detecting HCC were as follows: pooled sensitivity, 0.483 (95% confidence interval (CI) 0.459-0.507); pooled specificity, 0.929 (95% CI 0.916-0.940); DOR, 12.33 (95% CI 7.82-19.44); and area under the curve (AUC), 0.7564. CONCLUSIONS: AFP-L3% could be complementary to AFP as a marker for HCC.

The assessment of environmental pollution along the coast of Beibu Gulf, northern South China Sea: an integrated biomarker approach in the clam Meretrix meretrix.

The clam Meretrix meretrix was used as a biomonitor to implement an environmental monitoring program along the coast of Beibu Gulf in October 2011. This program not only analyzed biomarkers including acetylcholinesterase, glutathione peroxidase, glutathione S-transferase, catalase and superoxide dismutase activities, total glutathione content and lipid peroxidation level in M. meretrix but also adopted a multi-biomarker approach - integrated biomarker response (IBR) to assess the environmental quality in this ecosystem. In addition, the metal (Hg, As, Cu, Pb, Zn, Cd and Cr) and polychlorinated biphenyls (PCBs) content in the surface sediment at the study area were also measured. The results showed that IBR index was able to distinguish a space trend between sampling sites with different degrees of anthropogenic environmental stress. Integrated contamination degree were displayed in the form of star plots and compared to IBR plots. There was a visual consistency between the pollution level and IBR variation. Based on the results, it was proved that the IBR method coupled with chemical analysis was quite useful for the assessment of environmental pollution in the coastal system.