Transcriptome profiling reveals multiple regulatory pathways of Tamarix chinensis in response to salt stress.

KEY MESSAGE: Multiple regulatory pathways of T. chinensis to salt stress were identified through transcriptome data analysis. Tamarix chinensis (Tamarix chinensis Lour.) is a typical halophyte capable of completing its life cycle in soils with medium to high salinity. However, the mechanisms underlying its resistance to high salt stress are still largely unclear. In this study, transcriptome profiling analyses in different organs of T. chinensis plants in response to salt stress were carried out. A total number of 2280, 689, and 489 differentially expressed genes (DEGs) were, respectively, identified in roots, stems, and leaves, with more DEGs detected in roots than in stems and leaves. Gene Ontology (GO) term analysis revealed that they were significantly enriched in "biological processes" and "molecular functions". Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis revealed that "Beta-alanine metabolism" was the most differentially enriched pathway in roots, stems, and leaves. In pair-to-pair comparison of the most differentially enriched pathways, a total of 14 pathways, including 5 pathways in roots and leaves, 6 pathways in roots and stems, and 3 pathways in leaves and stems, were identified. Furthermore, genes encoding transcription factor, such as bHLH, bZIP, HD-Zip, MYB, NAC, WRKY, and genes associated with oxidative stress, starch and sucrose metabolism, and ion homeostasis, were differentially expressed with distinct organ specificity in roots, stems, and leaves. Our findings in this research provide a novel approach for exploring the salt tolerance mechanism of halophytes and identifying new gene targets for the genetic breeding of new plant cultivars with improved resistance to salt stress.

Clinical characteristics and survival prediction of surgical patients with invasive pancreatic cystic neoplasm: a large retrospective study over two decades.

PURPOSES: Invasive pancreatic cystic neoplasms (iPCNs) are an uncommon and biologically heterogeneous group of malignant neoplasms. We aimed to investigate the clinicopathological characteristics of iPCN patients and to develop nomograms for individual survival prediction after radical surgery. METHODS: Data of patients diagnosed with iPCN and pancreatic ductal adenocarcinoma (PDAC) between 2000 and 2018 from the SEER database were retrieved. The differences in clinical outcomes were evaluated using the Kaplan-Meier analysis. Nomograms were proposed based on the Cox regression model and internally validated by C-index, area under the curve (AUC) value, and calibration plot. RESULTS: A total of 7777 iPCN patients and 154,336 PDAC patients were enrolled. Most neoplasms were advanced, with 63.1% at stage IV. The 3-year overall survival (OS) and cancer-specific survival (CSS) rates in surgical patients were as follows: 45.7% and 50.1% for invasive intraductal papillary mucinous neoplasm (IPMN), 54.8% and 59.3% for invasive mucinous cystic neoplasm (MCN), 97.8% and 98.2% for invasive solid pseudopapillary neoplasm (SPN), 88.9% and 88.9% for invasive serous cystic neoplasm (SCN), and 27.3% and 30.5% for PDAC. Subgroup analyses showed no clinical benefit from chemotherapy or radiotherapy in lymph node-negative iPCN patients who underwent surgery. The following variables associated with OS and CSS were identified: age, race, chemotherapy, radiotherapy, histologic type, pathological grade, regional nodes examined, and T, N, and M stage. The nomograms had good discrimination and calibration by internal validation, with an AUC value of 0.800 for 3-year OS and 0.814 for 3-year CSS. CONCLUSION: Our study showed that the prognosis of iPCN patients was significantly better than PDAC patients. The proposed nomograms demonstrated substantially better discrimination and calibration.

Genome-wide analysis of growth-regulating factor genes in grape (Vitis vinifera L.): identification, characterization and their responsive expression to osmotic stress.

KEY MESSAGE: Identification, characterization and osmotic stress responsive expression of growth-regulating factor genes in grape. The growth and fruit production of grape vine are severely affected by adverse environmental conditions. Growth-regulating factors (GRFs) play a vital role in the regulation of plant growth, reproduction and stress tolerance. However, their biological functions in fruit vine crops are still largely unknown. In the present study, a total number of nine VvGRFs were identified in the grape genome. Phylogenetic and collinear relationship analysis revealed that they formed seven subfamilies, and have gone through three segmental duplication events. All VvGRFs were predicted to be nucleic localized and contained both the conserved QLQ and WRC domains at their N-terminals, one of the typical structural features of GRF proteins. Quantitative real-time PCR analyses demonstrated that all VvGRFs, with a predominant expression of VvGRF7, were constitutively expressed in roots, leaves and stems of grape plants, and showed responsive expression to osmotic stress. Further growth phenotypic analysis demonstrated that ectopic expression of VvGRF7 promoted the growth and sensitivity of transgenic Arabidopsis plants to osmotic stress. Our findings provide important information for the future study of VvGRF gene functions, and potential gene resources for the genetic breeding of new fruit vine varieties with improved fruit yield and stress tolerance.

Long non-coding RNA AFAP1-antisense RNA 1 promotes the proliferation, migration and invasion of gastric cancer cells and is associated with poor patient survival.

Gastric cancer (GC) is the second-leading cause of cancer-associated mortality worldwide. AFAP1-antisense RNA 1 (AFAP1-AS1), a long non-coding RNA (lncRNA), is believed to promote the aggressive progression of cancer; however, its role in GC remains largely unknown. In the present study, the expression of AFAP1-AS1 in GC tissues and cell lines was measured using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Knockdown of AFAP1-AS1 was performed using a lentiviral vector containing a short hairpin RNA. The proliferation of GC cells was measured using Cell Counting kit-8. The migration and invasion of GC cells were analyzed using a QCM Laminin Migration Assay kit and a Cell Invasion Assay kit. The levels of epithelial-mesenchymal transition (EMT)-associated proteins were detected by western blot analysis. The cut-off value of the expression of AFAP1-AS1 was evaluated using receiver operating characteristic (ROC) curves and patient survival rate was analyzed using Kaplan-Meier. The expression of AFAP1-AS1 was significantly increased in the primary tumor tissues of GC patients with lymph node metastasis or tumor node metastasis stage (stage III or IV; P<0.01). ROC curve analysis revealed that the expression of AFAP-AS1, at a cut-off value of 0.5040, could distinguish GC tissues from the matched normal tissues, with an AUC of 0.8802, sensitivity of 81.25% and specificity of 83.75%. The overexpression of AFAP1-AS1 was positively associated with the poor survival rates of GC patients. Furthermore, the downregulation of AFAP1-AS1 significantly inhibited the proliferation, migration and invasion of GC cells in vitro (P<0.01). The decrease in AFAP1-AS1 expression significantly suppressed the expression level of N-cadherin protein in GC cells and increased that of E-cadherin. The present study demonstrated that the expression signature of AFAP1-AS1 may serve as a biomarker for the diagnosis and prognosis of GC, and its downregulation may repress the aggressive progression of GC, partially through inhibiting the EMT progress.

Effective and persistent antitumor activity of HER2-directed CAR-T cells against gastric cancer cells in vitro and xenotransplanted tumors in vivo.

Human epidermal growth factor receptor 2 (HER2) proteins are overexpressed in a high proportion of gastric cancer (GC) cases and affect the maintenance of cancer stem cell (CSC) subpopulations, which are used as targets for the clinical treatment of patients with HER2-positive GC. Despite improvements in survival, numerous HER2-positive patients fail treatment with trastuzumab, highlighting the need for more effective therapies. In this study, we generated a novel type of genetically modified human T cells, expressing a chimeric antigen receptor (CAR), and targeting the GC cell antigen HER2, which harbors the CD137 and CD3ζ moieties. Our findings show that the expanded CAR-T cells, expressing an increased central memory phenotype, were activated by the specific recognition of HER2 antigens in an MHC-independent manner, and effectively killed patient-derived HER2-positive GC cells. In HER2-positive xenograft tumors, CAR-T cells exhibited considerably enhanced tumor inhibition ability, long-term survival, and homing to targets, compared with those of non-transduced T cells. The sphere-forming ability and in vivo tumorigenicity of patient-derived gastric cancer stem-like cells, expressing HER2 and the CD44 protein, were also inhibited. Our results support the future development and clinical application of this adoptive immunotherapy in patients with HER2-positive advanced GC.

Predictive factors for lymph node metastasis in early gastric cancer with signet ring cell histology: a meta-analysis.

BACKGROUND: Less invasive surgery is widely used in the treatment of early gastric cancer; however, no definite guidelines exist regarding indications for less invasive surgery to treat early gastric cancer with signet ring cell histology. The aim of this study was to identify risk factors for lymph node metastasis (LNM) in early signet ring cell carcinoma (SRC). An extensive search of PubMed, Embase and the Cochrane library was performed for pertinent articles involving early SRC and LNM. METHODS: Eligible data (gender, depth of invasion, lymphovascular invasion, size, ulceration, macroscopic type and location) were extracted from the included studies and systematically reviewed via a meta-analysis. Review Manager version 5.3 was used to perform the data processing. The Newcastle-Ottawa Scale was utilized to evaluate the quality of the included articles. RESULTS: Fourteen studies were included in the final analysis. After meta-analysis, female gender, submucosal invasion, lymphovascular invasion and size >20 mm were associated with LNM in early SRC. CONCLUSION: Four variables were identified as risk factors for LNM in early SRC. The significance of the results of the present study should be further confirmed in more early SRC patients for future clinical use.

A unified model of the hierarchical and stochastic theories of gastric cancer.

Gastric cancer (GC) is a life-threatening disease worldwide. Despite remarkable advances in treatments for GC, it is still fatal to many patients due to cancer progression, recurrence and metastasis. Regarding the development of novel therapeutic techniques, many studies have focused on the biological mechanisms that initiate tumours and cause treatment resistance. Tumours have traditionally been considered to result from somatic mutations, either via clonal evolution or through a stochastic model. However, emerging evidence has characterised tumours using a hierarchical organisational structure, with cancer stem cells (CSCs) at the apex. Both stochastic and hierarchical models are reasonable systems that have been hypothesised to describe tumour heterogeneity. Although each model alone inadequately explains tumour diversity, the two models can be integrated to provide a more comprehensive explanation. In this review, we discuss existing evidence supporting a unified model of gastric CSCs, including the regulatory mechanisms of this unified model in addition to the current status of stemness-related targeted therapy in GC patients.

A systematic review and meta-analysis of robot-assisted versus laparoscopically assisted gastrectomy for gastric cancer.

BACKGROUND: Robotic-assisted gastrectomy (RAG) has been used for gastric cancer since 2002. This meta-analysis was carried out to evaluate whether RAG is safer and more effective than conventional laparoscopically assisted gastrectomy (LAG) for gastric cancer. METHODS: We performed a manual search for these 2 types of operations (RAG and LAG) in the PubMed, Embase, and the Cochrane Library databases up to April 30, 2016. Twelve nonrandomized controlled trials that reported on RAG and LAG for gastric cancer were included. Outcomes evaluated included operation time, number of retrieved lymph nodes, blood loss, length of the resection margin, complications, and postoperative hospital stay. RESULTS: A total of 3744 patients in 12 studies were included (1134 patients in the RAG group and 2610 patients in the LAG group). The operation time was significantly shorter in the LAG group [weighted mean difference (WMD) 42.0 (95% confidence interval, 95% CI 28.11-55.89) minutes; P < .00001], while the loss of blood volume was lower in the RAG group (P = .01). The number of retrieved lymph nodes, duration of postoperative stay, length of the proximal resection margin, length of the distal resection margin, and postoperative complications were similar between groups. CONCLUSION: We conclude that RAG is a safe and appropriate treatment for gastric cancer patients in comparison to LAG. Nevertheless, RAG is not superior to LAG. Future research on RAG should focus on comparing the differences in retrieved lymph nodes in different tiers, evaluating the postoperative recovery and reducing the cost of the treatment.

Expression of decorin in intestinal tissues of mice with inflammatory bowel disease and its correlation with autophagy.

The aim of this study was to investigate the expression of decorin (DCN) in the intestinal tissues of mice with inflammatory bowel disease (IBD) and its correlation with autophagy. The IBD mouse model was created by intrarectal injection of trinitrobenzene sulfonic acid. The pathology of colon tissues of the mice was examined using hematoxylin and eosin staining. Expression of DCN and the proteins associated with autophagy was detected using immunohistochemistry. Normal human colon mucosal epithelial cells (NCM460 cells) were transfected with DCN expression plasmid and the expression of DCN and autophagy-associated proteins was detected by western blot analysis. Cell apoptosis was studied using an Annexin V apoptosis detection assay and intracellular autophagosomes were observed using electron microscopy. The IBD mouse model was successfully established. Thickening, edema and inflammatory cell infiltration of the intestinal wall were observed in the IBD mice. The expression of DCN as well as the autophagy-associated proteins beclin 1 and LC3B, was increased in the intestinal tissues of the IBD mice. Furthermore, in the NCM460 cells transfected with DCN, the expression of beclin 1 and LC3B was upregulated, while p62 expression was downregulated. Intracellular autophagosomes were increased and apoptosis was decreased in the cells with DCN overexpression. Inhibition of autophagy reversed the effects of DCN on apoptosis. Therefore, DCN is able to induce autophagy and protect intestinal cells during the occurrence and development of IBD.

[Application of synchrotron radiation X-ray fluorescence to investigating the distribution of trace elements in different organs of greenhouse rape].

The trace elements distribution in the root and leaf of greenhouse rape was determined with synchrotron radiation XRF, and many essential elements for human health were found, including Co, Mn, Ni, Fe, Cu, Zn, K and Ca, among which the contents of Ca, Mn, Fe, Cu and Zn are especially higher than the other elements. The results proved that the contents of different elements in different parts of leaves and roots were inequable. Generally, the contents of K, Ca and Mg in the roots are significantly higher than those in leaves, while the contents of Fe, Co, Ni, Cu and Zn in the leaves are apparently higher than those in roots. And the contents of all the elements in lateral roots are much lower. The contents of K, Ca and Co in the central section of leaf were higher than those in the edge, and the contents are gradually increasing from the edge to center; the contents of Cu, Mn, Fe and Ni in the tip of leaf were higher than those in base, and it increases gradually from the base to the tip of leaf; the content of Zn in the leaf is absolutely uniform; all the elements in the middle of root were higher than those in two ends, and the contents in the base of root are commonly higher than those in the tip of root.