White spot syndrome virus entry is dependent on multiple endocytic routes and strongly facilitated by Cq-GABARAP in a CME-dependent manner.

White spot syndrome virus (WSSV) is a lethal pathogen of shrimp and many other crustaceans, including crayfish. However, the molecular mechanism underlying its cellular entry remains elusive due to the lack of shrimp cell lines for viral propagation. Crayfish hematopoietic tissue (Hpt) cell culture was recently established as a good model for WSSV infection study. Here, we showed that multiple endocytic routes, including clathrin-mediated endocytosis (CME), macropinocytosis and caveolae-mediated endocytosis, were indispensably employed for the viral entry into Hpt cell of the crayfish Cherax quadricarinatus. Intriguingly, cellular autophagic activity was positively correlated with efficient viral entry, in which a key autophagy-related protein, γ-aminobutyric acid receptor-associated protein (Cq-GABARAP), that not only localized but also co-localized with WSSV on the Hpt cell membrane, strongly facilitated WSSV entry by binding to the viral envelope VP28 in a CME-dependent manner that was negatively regulated by Cq-Rac1. Furthermore, cytoskeletal components, including Cq-ß-tubulin and Cq-ß-actin, bound to both recombinant rCq-GABARAP and WSSV envelope proteins, which likely led to viral entry promotion via cooperation with rCq-GABARAP. Even under conditions that promoted viral entry, rCq-GABARAP significantly reduced viral replication at an early stage of infection, which was probably caused by the formation of WSSV aggregates in the cytoplasm.

Detrimental effect of CO2-driven seawater acidification on a crustacean brine shrimp, Artemia sinica.

The effects of the decline in ocean pH, termed as ocean acidification due to the elevated carbon dioxide in the atmosphere, on calcifying organisms such as marine crustacean are unclear. To understand the possible effects of ocean acidification on the physiological responses of a marine model crustacean brine shrimp, Artemia sinica, three groups of the cysts or animals were raised at different pH levels (8.2 as control; 7.8 and 7.6 as acidification stress according to the predictions for the end of this century and next century accordingly) for 24 h or two weeks, respectively, followed by examination of their hatching success, morphological appearance such as deformity and microstructure of animal body, growth (i.e. body length), survival rate, expression of selected genes (involved in development, immunity and cellular activity etc), and biological activity of several key enzymes (participated in antioxidant responses and physiological reactions etc). Our results clearly demonstrated that the cysts hatching rate, growth at late stage of acidification stress, and animal survival rate of brine shrimp were all reduced due to lower pH level (7.6 & 7.8) on comparison to the control group (pH 8.2), but no obvious change in deformity or microstructure of brine shrimp was present under these acidification stress by microscopy observation and section analysis. In addition, the animals subjected to a lower pH level of seawater underwent changes on their gene expressions, including Spätzle, MyD88, Notch, Gram-negative bacteria binding protein, prophenoloxidase, Apoptosis inhibitor 5, Trachealess, Caveolin-1 and Cyclin K. Meanwhile, several key enzyme activities, including superoxide dismutase, catalase, peroxidase, alkaline phosphatase and acid phosphatase, were also affected by acidified seawater stress. Taken together, our findings supports the idea that CO2-driven seawater acidification indeed has a detrimental effect, in case of hatching success, growth and survival, on a model crustacean brine shrimp, which will increase the risk of juvenile brine shrimp and possibly also other crustaceans, as important live feeds for aquaculture being introduced in the ecosystem especially the marine food webs.

Identification of a serine proteinase homolog (Sp-SPH) involved in immune defense in the mud crab Scylla paramamosain.

Clip domain serine proteinase homologs are involved in many biological processes including immune response. To identify the immune function of a serine proteinase homolog (Sp-SPH), originally isolated from hemocytes of the mud crab, Scylla paramamosain, the Sp-SPH was expressed recombinantly and purified for further studies. It was found that the Sp-SPH protein could bind to a number of bacteria (including Aeromonas hydrophila, Escherichia coli, Staphylococcus aureus, Vibrio fluvialis, Vibrio harveyi and Vibrio parahemolyticus), bacterial cell wall components such as lipopolysaccharide or peptidoglycan (PGN), and ß-1, 3-glucan of fungus. But no direct antibacterial activity of Sp-SPH protein was shown by using minimum inhibitory concentration or minimum bactericidal concentration assays. Nevertheless, the Sp-SPH protein was found to significantly enhance the crab hemocyte adhesion activity (paired t-test, P<0.05), and increase phenoloxidase activity if triggered by PGN in vitro (paired t-test, P<0.05). Importantly, the Sp-SPH protein was demonstrated to promote the survival rate of the animals after challenge with A. hydrophila or V. parahemolyticus which were both recognized by Sp-SPH protein, if pre-incubated with Sp-SPH protein, respectively. Whereas, the crabs died much faster when challenged with Vibrio alginolyiicus, a pathogenic bacterium not recognized by Sp-SPH protein, compared to those of crabs challenged with A. hydrophila or V. parahemolyticus when pre-coated with Sp-SPH protein. Taken together, these data suggested that Sp-SPH molecule might play an important role in immune defense against bacterial infection in the mud crab S. paramamosain.

Growth response of Zea mays L. in pyrene-copper co-contaminated soil and the fate of pollutants.

Phytoremediation, use of plants for remediation, is an emerging technology for treating heavy metals or a final polishing step for the high-level organic contamination, and may be suitable for remediation of heavy metal and organic co-contaminated soil. The aim of this study was to investigate the influence of co-contamination on the growth of Zea mays L. and the fate of both heavy metal and organic pollutants, using Cu and pyrene as the model pollutants. Results showed that shoot and root biomass were affected by the copper-pyrene co-contamination, although maize grown in spiked soils showed no outward signs of phytotoxicity. With the initial concentration of 50,100 and 500 mg/kg, pyrene tended to alleviate the inhibition of Cu to Z. mays L. Pyrene in both planted and non-planted soil was greatly decreased at the end of the 4-week culture, accounting for 16-18% of initial extractable concentrations in non-planted soil and 9-14% in planted soil, which indicated that the dissipation of soil pyrene was enhanced in the presence of vegetation probably due to the biodegradation and association with the soil matrix. With the increment of Cu level, residual pyrene in the planted soil tended to increase. The pyrene residual in the presence of high concentration of Cu was even higher in the planted soil than that in the non-planted soil, which suggested that the change of the microbial composition and microbial activity or the modified root physiology under Cu stress was probably unbeneficial to the dissipation of pyrene. A more thorough understanding of the mechanisms by which metals affect the dissipation of organic pollutants in the rhizosphere could provide a much better framework on which to base manipulation. Unlike pyrene, heavy metal copper cannot be degraded. Decontamination of Cu from contaminated soils in this system required the removal of Cu by plants. It was observed that the ability of Cu phytoextraction would be inhibited under co-contamination of high level of pyrene in highly Cu-polluted soil. In the treatment of 400 mg Cu/kg and 500 mg pyrene/kg, the accumulation of Cu was less than half of that in 400 mg Cu/kg treatment.

Partial glossectomy assisted with temperature-controlled radiofrequency for treating macroglossia / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the clinical efficacy of partial glossectomy assisted with temperature-controlled radiofrequency for treating macroglossia.</p><p><b>METHODS</b>There were 4 patients performed this procedure. We took a rhombus shape incision in the middle of the tongue and performed a wedge excision.</p><p><b>RESULTS</b>The mouth can close entirely in all of patients and there weren' t hemorrhage and obviously swollen; Tongue's sensory function hadn't disturbance. Masticate function were normal. One patient still had the symptom that tongue lied outside the oral cavity occasionally after operation. The symptom was disappeared after Temperature-controlled radiofrequency (TCRF) ablation. All of patients' parent were satisfied with the results.</p><p><b>CONCLUSIONS</b>Partial glossectomy assisted with temperature-controlled radiofrequency for treating macroglossia is an effective, much safer and less invasive procedure without obvious adverse reactions. There are better prospects for applying.</p>

Repair of eyelid defect with a prefabricated flap / 中华整形外科杂志

<p><b>OBJECTIVE</b>To introduce a new method for repair of eyelid defects.</p><p><b>METHODS</b>Eyelid reconstruction was performed using a prefabricated island flap with an expanded forehead myocutaneous flap and hard palate mucoperiosteal graft.</p><p><b>RESULTS</b>Five patients with congenital or acquired eyelid defects were successfully treated with this method.</p><p><b>CONCLUSION</b>The introduced method is rational and practical for clinical applications.</p>