Comparative proteomics of three Chinese potato cultivars to improve understanding of potato molecular response to late blight disease.

BACKGROUND: Late blight disease (LBD) caused by the pathogen Phytophthora infestans (PI), is the most devastating disease limiting potato (Solanum tuberosum) production globally. Currently, this disease pathogen is re-emerging and appearing in new areas at a very high intensity. A better understanding of the natural defense mechanisms against PI in different potato cultivars especially at the protein level is still lacking. Therefore, to elucidate potato proteome response to PI, we investigated changes in the proteome and leaf morphology of three potato cultivars, namely; Favorita (FA), Mira (MA), and E-malingshu N0.14 (E14) infected with PI by using the iTRAQ-based quantitative proteomics analysis. RESULTS: A total of 3306 proteins were found in the three potato genotypes, and 2044 proteins were quantified. Cluster analysis revealed MA and E14 clustered together separately from FA. The protein profile and related functions revealed that the cultivars shared a typical hypersensitive response to PI, including induction of elicitors, oxidative burst, and suppression of photosynthesis in the potato leaves. Meanwhile, MA and E14 deployed additional specific response mechanism different from FA, involving high induction of protease inhibitors, serine/threonine kinases, terpenoid, hormone signaling, and transport, which contributed to MA tolerance of LBD. Furthermore, inductions of pathogenesis-related proteins, LRR receptor-like kinases, mitogen-activated protein kinase, WRKY transcription factors, jasmonic acid, and phenolic compounds mediate E14 resistance against LBD. These proteins were confirmed at the transcription level by a quantitative polymerase chain reaction and at the translation level by western-blot. CONCLUSIONS: We found several proteins that were differentially abundant among the cultivars, that includes common and cultivar specific proteins which highlighted similarities and significant differences between FA, MA, and E14 in terms of their defense response to PI. Here the specific accumulation of mitogen-activated protein kinase, Serine/threonine kinases, WRKY transcription played a positive role in E14 immunity against PI. The candidate proteins identified reported in this study will form the basis of future studies and may improve our understanding of the molecular mechanisms of late blight disease resistance in potato.

Quantitative Proteomics of Potato Leaves Infected with Phytophthora infestans Provides Insights into Coordinated and Altered Protein Expression during Early and Late Disease Stages.

In order to get a better understanding of protein association during Solanum tuberosum (cv. Sarpo Mira)⁻Phytophthora infestans incompatible interaction, we investigated the proteome dynamics of cv. Sarpo Mira, after foliar application of zoospore suspension from P. infestans isolate, at three key time-points: zero hours post inoculation (hpi) (Control), 48 hpi (EI), and 120 hpi (LI); divided into early and late disease stages by the tandem mass tagging (TMT) method. A total of 1229 differentially-expressed proteins (DEPs) were identified in cv. Sarpo Mira in a pairwise comparison of the two disease stages, including commonly shared DEPs, specific DEPs in early and late disease stages, respectively. Over 80% of the changes in protein abundance were up-regulated in the early stages of infection, whereas more DEPs (61%) were down-regulated in the later disease stage. Expression patterns, functional category, and enrichment tests highlighted significant coordination and enrichment of cell wall-associated defense response proteins during the early stage of infection. The late stage was characterized by a cellular protein modification process, membrane protein complex formation, and cell death induction. These results, together with phenotypic observations, provide further insight into the molecular mechanism of P. infestans resistance in potatos.

Human acellular amniotic membrane incorporating exosomes from adipose-derived mesenchymal stem cells promotes diabetic wound healing.

BACKGROUND: Diabetic wounds threaten the health and quality of life of patients and their treatment remains challenging. ADSC-derived exosomes have shown encouraging results in enhancing diabetic wound healing. However, how to use exosomes in wound treatment effectively is a problem that needs to be addressed at present. METHODS: A diabetic mouse skin wound model was established. ADSC-derived exosomes (ADSC-Exos) were isolated, and in vitro application of exosomes was evaluated using human umbilical vein endothelial cells (HUVECs) and human dermal fibroblasts (HDFs). After preparation and characterization of a scaffold of human acellular amniotic membrane (hAAM) loaded with ADSC-Exos in vitro, they were transplanted into wounds in vivo and wound healing phenomena were observed by histological and immunohistochemical analyses to identify the wound healing mechanism of the exosome-hAAM composites. RESULTS: The hAAM scaffold dressing was very suitable for the delivery of exosomes. ADSC-Exos enhanced the proliferation and migration of HDFs and promoted proliferation and tube formation of HUVECs in vitro. In vivo results from a diabetic skin wound model showed that the hAAM-Exos dressing accelerated wound healing by regulating inflammation, stimulating vascularization, and promoting the production of extracellular matrix. CONCLUSION: Exosome-incorporated hAAM scaffolds showed great potential in promoting diabetic skin wound healing, while also providing strong evidence for the future clinical applications of ADSC-derived exosomes.

Effect of radical resection combined with antiviral therapy in patients with hepatitis B virus-associated hepatocellular carcinoma and prognostic analysis.

PURPOSE: To observe the clinical effect of radical resection combined with antiviral therapy in patients with hepatitis B virus (HBV)-associated hepatocellular carcinoma (HCC), and to analyze the risk factors affecting its prognosis. METHODS: The clinical data of 132 patients with HBV-associated HCC treated in our hospital from January 2015 to December 2016 were retrospectively analyzed, and the patients were randomly divided into Control group (n=66) and Anti-virus group (n=66). The changes in liver function indexes, HBV-deoxyribonucleic acid (DNA) load and alpha fetoprotein (AFP) level were compared between the two groups before and after treatment. The tumor recurrence and patients' survival were recorded during the follow-up period, and the possible influencing factors for the prognosis of patients with HBV-associated HCC were analyzed. RESULTS: After treatment, the levels of alanine aminotransferase (ALT), albumin (ALB), prealbumin (PA) and AFP significantly declined in both groups (p<0.05), while the levels of ALT, PA and AFP were significantly lower in Anti-virus group than those in Control group (p<0.001). After treatment, the HBV-DNA level declined in both groups compared with that before treatment, while it was obviously lower in Anti-virus group than that in Control group (p<0.001). During treatment, the total incidence rate of complications in Anti-virus group was 37.9%, markedly lower than that in Control group 59.1% (p=0.023). The results of log-rank test showed that both OS and PFS rates were far higher in Anti-virus group than those in Control group (p=0.043, p=0.034). The results of Cox multivariate analysis revealed that a low degree of tumor histological differentiation, a large diameter of tumor and no antiviral therapy were independent risk factors affecting the OS rate of patients after treatment (p=0.030, p=0.017). CONCLUSIONS: Antiviral therapy after radical resection of HBV-associated HCC can effectively inhibit the replication of HBV, reduce the recurrence rate of tumor, and prolong the OS of patients. Low grade of tumor histological differentiation, large diameter of tumor and no antiviral therapy are independent risk factors affecting the OS rate of patients after treatment.