Correction: FoQDE2-dependent milRNA promotes Fusarium oxysporum f. sp. cubense virulence by silencing a glycosyl hydrolase coding gene expression.

[This corrects the article DOI: 10.1371/journal.ppat.1010157.].

Asynchronous control for semi-Markov switching systems with singular perturbation and an improved protocol.

This study addresses the asynchronous control problem for a semi-Markov switching system in the presence of singular perturbation and an improved triggering protocol. To decrease the occupation of network resources, an improved protocol is skillfully established by adopting two auxiliary offset variables. Unlike the existing protocols, the established improved protocol is capable of arranging information transmission with more degrees of freedom, thereby reducing the communication frequency and maintaining control performance. Apart from the reported hidden Markov model, a nonhomogeneous hidden semi-Markov model is used to handle the mode mismatch between the systems and controllers. Benefiting from Lyapunov techniques, parameter-dependent sufficient conditions are devised to ensure a stochastically stable subject to a predetermined performance. Finally, the validity and practicability of the theoretical results are verified via a numerical example and a tunnel diode circuit model.

A Dynamic-Memory Event-Triggered Protocol to Multiarea Power Systems With Semi-Markov Jumping Parameter.

This work deals with the dynamic-memory event-triggered-based load frequency control issue for interconnected multiarea power systems (IMAPSs) associated with random abrupt variations and deception attacks. To facilitate the transient faults, a semi-Markov process is addressed to model the dynamic behavior of IMAPSs. In order to modulate transmission frequency, a novel area-dependent dynamic-memory event-triggered protocol (DMETP) is scheduled by resorting to a set of the historically released packets (HRPs), which ensures better dynamic performance. From the viewpoint of the defender, the randomly occurring deception attack is taken into account, which is regulated by a Bernoulli-distributed scalar. Benefitting from the DMETP scheduling, a novel framework of the memory-based asynchronous control strategy is formulated, in which the hidden semi-Markov model is adopted to reveal the mode mismatches. Based on the Lyapunov theory, sufficient conditions are established to ensure the stochastic stability of the resulting systems. In the end, the simulation result is presented to reveal the efficiency of the proposed dynamic-memory event-triggered-based approach.

FoQDE2-dependent milRNA promotes Fusarium oxysporum f. sp. cubense virulence by silencing a glycosyl hydrolase coding gene expression.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate protein-coding gene expression primarily found in plants and animals. Fungi produce microRNA-like RNAs (milRNAs) that are structurally similar to miRNAs and functionally important in various biological processes. The fungus Fusarium oxysporum f. sp. cubense (Foc) is the causal agent of Banana Fusarium vascular wilt that threatens global banana production. It remains uncharacterized about the biosynthesis and functions of milRNAs in Foc. In this study, we investigated the biological function of milRNAs contributing to Foc pathogenesis. Within 24 hours post infecting the host, the Argonaute coding gene FoQDE2, and two Dicer coding genes FoDCL1 and FoDCL2, all of which are involved in milRNA biosynthesis, were significantly induced. FoQDE2 deletion mutant exhibited decreased virulence, suggesting the involvement of milRNA biosynthesis in the Foc pathogenesis. By small RNA sequencing, we identified 364 small RNA-producing loci in the Foc genome, 25 of which were significantly down-regulated in the FoQDE2 deletion mutant, from which milR-87 was verified as a FoQDE2-depedent milRNA based on qRT-PCR and Northern blot analysis. Compared to the wild-type, the deletion mutant of milR-87 was significantly reduced in virulence, while overexpression of milR-87 enhanced disease severity, confirming that milR-87 is crucial for Foc virulence in the infection process. We furthermore identified FOIG_15013 (a glycosyl hydrolase-coding gene) as the direct target of milR-87 based on the expression of FOIG_15013-GFP fusion protein. The FOIG_15013 deletion mutant displayed similar phenotypes as the overexpression of milR-87, with a dramatic increase in the growth, conidiation and virulence. Transient expression of FOIG_15013 in Nicotiana benthamiana leaves activates the host defense responses. Collectively, this study documents the involvement of milRNAs in the manifestation of the devastating fungal disease in banana, and demonstrates the importance of milRNAs in the pathogenesis and other biological processes. Further analyses of the biosynthesis and expression regulation of fungal milRNAs may offer a novel strategy to combat devastating fungal diseases.

Inhibition of Murine Breast Cancer Metastases by Hydrophilic As4S4 Nanoparticles Is Associated With Decreased ROS and HIF-1α Downregulation.

Arsenic sulfide (As4S4) is a mineral drug that can be administrated orally and has been applied in the treatment of myeloid leukemia. The aim of this work is to investigate the therapeutic effect of As4S4 in highly metastatic triple-negative breast cancer (TNBC) animal model, as As4S4 has not been applied in the treatment of breast cancer yet. To overcome the poor solubility of original As4S4, a formulation of hydrophilic As4S4 nanoparticles (e-As4S4) developed previously was applied to mouse breast cancer cells as well as the tumor-bearing mice. It was shown that e-As4S4 was much more cytotoxic than r-As4S4, strongly inhibiting the proliferation of the cells and scavenging intracellular reactive oxygen species (ROS). The oral administration of e-As4S4 significantly increased the accumulation of arsenic in the tumor tissue and eliminated ROS in tumor tissues. Besides, e-As4S4 could also inhibit the activation of hypoxia-inducible factor-1α (HIF-1α) and NLRP3 inflammasomes. Consequently, the angiogenesis was reduced, the metastasis to lung and liver was inhibited and the survival of tumor-bearing mice was prolonged. In conclusion, e-As4S4 holds great potential for an alternative therapeutics in the treatment of breast cancer, due to its unique function of correcting the aggressive microenvironment.

Cationic polysaccharide spermine-pullulan drives tumor associated macrophage towards M1 phenotype to inhibit tumor progression.

Macrophages are predominant immune cells in the tumor microenvironment where they display an immunosuppressive M2 phenotype to support tumor growth. Reprogramming M2-like tumor-associated macrophages (TAMs) to antitumor M1 phenotype represents as a promising strategy in cancer immunotherapy. In this work we reported that one cationic polysaccharide spermine modified pullulan (PS) could act as an effective immunological stimulator to modulating either naïve (M0) or M2 macrophages towards M1 phenotype. We showed that PS upregulated the expression of TLR1/3/4 and promoted the phosphorylation of Akt, Erk, JNK, following the activation of NF-κB, which led to the polarization towards M1. In a murine breast cancer model of tumor cell 4T1 inoculation, subcutaneous injection of PS induced effective antitumor effect through reprogramming M2 macrophages in the tumor microenvironment to M1, increased CD4+ and CD8+ T cells, and decreased the expression of CD31 in the tumor mass, which together inhibited the tumor progression and the metastasis in lung and liver, leading to the prolong of the mice survival. In conclusion, PS could effectively stimulate the immunological function of macrophages. Therefore, PS may provide a novel immunological stimulator to cancer immune therapies.

Ratio of Polycation and Serum Is a Crucial Index for Determining the RNAi Efficiency of Polyplexes.

We report that the mass ratio of the polycation to serum in the medium determines the (RNA interference) RNAi efficiency in vitro by using spermine-modified pullulan (Ps) and spermine-modified dextran (Ds) as polycation models. The high ratio of Ps to serum protein (Ps/Pr) mediated the formation of larger polyplexes, which led to the promoted cellular uptake, enhanced lysosomal escape, and elevated RNAi efficiency. In addition, the supplementary of free Ps also enhanced small interfering RNA transfection because of the elevation of Ps/Pr. Similar results were obtained with Ds. Compared with the adjustment of the nitrogen to phosphate (N/P) ratio in the polyplex, these findings revealed a more applicable strategy to tune the polycation-mediated RNAi efficiency in the serum-containing culture medium.

Visible light-switched cytosol release of siRNA by amphiphilic fullerene derivative to enhance RNAi efficacy in vitro and in vivo.

Cationic macromolecules are attractive for use as small interfering RNA (siRNA) carriers due to their performance in non-immunological reactions, customization during synthesis, and low costs compared to viral carriers. However, their low transfection efficiency substantially hinders their application in both clinical practices and academic research, which is mostly attributable to the low capacity of siRNA/cationic macromolecule complexes to escape lysosomes. To address this challenge, we designed an amphiphilic fullerene derivative (C60-Dex-NH2) for efficient and controllable siRNA delivery. To synthesize C60-Dex-NH2, terminally aminated dextran was conjugated to C60. The conjugate was further cationized by covalently introducing ethylenediamine to the dextran. The physicochemical characteristics of C60-Dex-NH2 was examined with elemental analyses, gel permeation chromatography, solid-state nuclear magnetic resonance (13C, HPDEC), agarose gel electrophoresis, and dynamic light scattering. The cytotoxicity, cellular uptake, intracellular distribution, and in vitro RNA interference (RNAi) of siRNA/C60-Dex-NH2 complex was evaluated in the human breast cancer cell line MDA-MB-231. The RNAi efficiencies mediated by C60-Dex-NH2in vivo was evaluated in subcutaneous tumor-bearing mice. The results showed that C60-Dex-NH2 has a specific amphiphilic skeleton and could form micelle-like aggregate structures in water, which could prevent siRNA from destroying by reactive oxygen species (ROS). When exposed to visible light, C60-Dex-NH2 could trigger controllable ROS generation which could destroy the lysosome membrane, promote the lysosomal escape, and enhance the gene silencing efficiency of siRNA in vitro and in vivo. The gene silencing efficiency could reach a maximum of 53% in the MDA-MB-231-EGFP cells and 69% in the 4T1-GFP-Luc2 tumor-bearing mice. STATEMENT OF SIGNIFICANCE: We designed a novel photosensitive amphiphilic carrier (C60-Dex-NH2) for efficient and controllable siRNA delivery, which can be used in gene therapy. We showed that C60-Dex-NH2 could destroy lysosome membrane via controllable generation of ROS when exposed to light, which can help siRNA to escape from lysosome before degradation. This can enhance the gene silencing efficiency significantly and provides a useful way to regulate RNAi efficiency by light. One advantage for C60-Dex-NH2 system is C60 has broad absorbance spectrum and can be activated by weak visible light; Furthermore, C60-Dex-NH2 has a specific amphiphilic structure, which may prevent siRNA from degrading and allows C60-Dex-NH2 to embed into the lipid membrane of lysosome to improve the ROS induced lysosomal disturbance after internalization.

[Isolation and Identification of Bacteria and Mycete from Preservation of Morinda officinalis].

OBJECTIVE: In order to select antistaling agent which had obvious antibacterial effect on Morinda officinalis, the colony counts of microorganism from the surface of Morinda officinalis and the species of bacteria and mycete were analyzed. METHODS: Bacteria and moulds classification identification method were carried out. RESULTS: After using 10 g burdock oligosaccharide, 20 g chitosan and 0. 5 g potassium sorbate treated for 60 d, the total bacteria on the surface of Morinda officinalis was 3. 0 x 10(4) cfu/g, mycete total of 1. 5 x 10(4) cfu/g, preservation effect was better than the other treatments. CONCLUSION: The common advantage microorganisms group of Morinda officinalis on the surface are from Leuconostoc ,Aerococcus and Penicillium genera.

Inhibition of α-smooth muscle actin expression and migration of pterygium fibroblasts by coculture with amniotic mesenchymal stem cells.

PURPOSE: Epithelial-mesenchymal transition (EMT) of normal conjunctival tissues is a major reason of pterygium generation; α-smooth muscle actin (α-SMA) is a maker in EMT. In this study, we investigated if human amniotic mesenchymal stem cells (hAMSCs) can inhibit α-SMA expression and migration of human pterygium fibroblasts (hPFs) in vitro. MATERIALS AND METHODS: Coculture of hAMSCs and hPFs was completed by using a Transwell coculture system. The α-SMA expression of hPFs was detected by immunocytochemistry and western blot analysis. The migration ability of hPFs was measured by means of a migration assay. Immunoreactivity for α-SMA was detected in all pterygium fibroblasts examined. RESULTS: The expression of α-SMA in cocultured hPFs was significantly lower than in hPFs. Similar result was demonstrated in western blot analysis. In addition, migration assay showed that hAMSCs reduce the migration of hPFs. CONCLUSIONS: These results suggested that hAMSCs have the potential to inhibit the generation and invasiveness of pterygium in vitro.

[Application of surgical robotic system in patients with gastric cancer: a report of 120 cases].

OBJECTIVE: To evaluate the safety, feasibility, and efficacy of robotic gastrectomy for gastric cancer using da Vinci surgical system. METHODS: A total of 120 patients who underwent robotic gastrectomy using da Vinci surgical system for gastric cancer from May 2010 to April 2012. Data regarding surgical and early oncological outcomes were systematically collected in a specific database for statistical analysis. RESULTS: There were 74 males and 46 females, with a mean age of 58.5 (22-80) years old. The type of robotic operation included 62 distal subtotal gastrectomies, 35 total gastrectomies, and 23 proximal gastrectomies. Reconstruction of the alimentary tract was achieved using extracorporeal method through a minilaparotomy in 55 cases, and intracorporeal robot-sewn anastomosis in 65 cases. There was 1 conversion (0.9%). The operative time was (245±50) min and the docking time (17±5) min. The blood loss was (70±45) ml. The number of harvested lymph nodes was 22.5±10.7. The resection margins were negative in all surgical specimens. The postoperative pathological stage consisted of stage I B in 24 cases, stage II in 28, stage III A in 47, and stage III B in 21. Six patients (5%) developed postoperative complication including 1 case of duodenal stump leakage, 2 cases of esophagus-gastric leakage, 1 case of small bowel obstruction, 1 case of delayed gastric emptying, and 1 case of abdominal bleeding. All the complications were cured by reoperation or conservative therapy, and there were no postoperative 30-day deaths. The postoperative hospital stay was (6.3±2.6) days. CONCLUSION: Robotic gastrectomy for the treatment of gastric cancer is a feasible and safe procedure, and is advantageous in terms of lymphadenectomy and alimentary tract reconstruction.

[Robotic-assisted laparoscopic colectomy for colon cancer: a report of 13 cases].

OBJECTIVE: To investigate the safety and feasibility of robotic-assisted laparoscopic colectomy for colonic cancer. METHODS: The clinical outcomes of 13 patients with colon cancer undergoing robotic-assisted laparoscopic colectomy from May 2010 to November 2010 were retrospectively evaluated. RESULTS: All the operations were performed successfully, including 5 right colectomies, 3 left colectomies, and 5 sigmoidectomies. The operative time was (171.5±31.8) minutes. The estimated blood loss was (54.6±21.8) ml. Time to the return of bowel function was (60.9±15.8) hours and postoperative hospital stay was (6.4±3.6) days. There was one patient developed fat liquefaction at the incision. No bleeding, anastomotic fistula, anastomotic stenosis, or other complications were found. CONCLUSION: Robotic-assisted laparoscopic colectomy is safe and feasible for colon cancer resection.

Fast-track rehabilitation program vs conventional care after colorectal resection: a randomized clinical trial.

AIM: To compare the fast-track rehabilitation program and conventional care for patients after resection of colorectal cancer. METHODS: One hundred and six consecutive patients who underwent fast-track rehabilitation program were encouraged to have early oral feeding and movement for early discharge, while 104 consecutive patients underwent conventional care after resection of colorectal cancer. Their gastrointestinal functions, postoperative complications and hospital stay time were recorded. RESULTS: The restoration time of gastrointestinal functions in the patients was significantly faster after fast-track rehabilitation program than after conventional care (2.1 d vs 3.2 d, P<0.01). The percentage of patients who developed complications was significantly lower 30 d after fast-track rehabilitation program than after conventional care (13.2% vs 26.9%, P<0.05). Also, the percentage of patients who had general complications was significantly lower 30 d after fast-track rehabilitation program than after conventional care (6.6% vs 15.4%, P<0.05). The postoperative hospital stay time of the patients was shorter after fast-track rehabilitation program than after conventional care (5 d vs 7 d, P<0.01). No significant difference was observed in the re-admission rate 30 d after fast-track rehabilitation program and conventional care (3.8% vs 8.7%). CONCLUSION: The fast-track rehabilitation program can significantly decrease the complications and shorten the time of postoperative hospital stay of patients after resection colorectal cancer.

[Clinical use of fast-track surgery in colorectal cancer patients:report of 116 cases].

OBJECTIVE: To investigate the efficacy and safety of fast-track surgery(FTS) in patients with colorectal cancer. METHODS: One hundred and sixteen patients with colorectal cancer underwent colorectal operation between June 2006 and May 2008. FTS procedures were used in these patients during the study period. Data were retrospectively analyzed pertaining to post-operative hospital stay and operative complications. RESULTS: The median postoperative hospital stay was (5.6+/-5.4)(range 1-54) d. Fourteen patients(12.1%) had postoperative complications, in which the rate of anastomotic leakage and wound infection was 1.7% and 2.6% respectively. 30-day readmission rate was 1.7%, and the 30-day mortality was 0.9%. CONCLUSION: Fast-track surgery is feasible in an unselected patient population undergoing elective colorectal cancer resection without compromising surgical outcomes.