Filamentous fungi can secrete large amounts of proteins, glycosylate proteins and grow faster than plant, insect or mammalian cells. With the advances in fungal molecular genetics, strain improvement, and especially fungal genomics, filamentous fungi are developed as microbial cell factories for the production of heterologous proteins. This review focuses on recent developments of filamentous fungi as production hosts, such as protein quality control mechanisms, the secretion pathways, protein modification, strain stability, and most importantly the application of functional genomics in protein expression. At the same time, the strategies for improving heterologous protein production were also discussed in details.
Biotechnology ; methods ; trends ; Cloning, Molecular ; Fermentation ; genetics ; physiology ; Fungi ; genetics ; metabolism ; Genetic Engineering ; Protein Processing, Post-Translational ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To evaluate the role of spinal Leucine-rich Repeat Kinase 2 (LRRK2) in neuropathic pain in rats.Methods:Fifty SPF healthy male Sprague-Dawley rats, aged 6-7 weeks, weighing 210-245 g, were divided into 5 groups ( n=10 each) using a random number table method: control group (C group), neuropathic pain group (NP group), low dose GNE-7915 group (low-dose GNE-7915 group), medium-dose GNE-7915 group (medium-dose GNE-7915 group), and high-dose GNE-7915 group (high-dose GNE-7915 group). Neuropathic pain was induced by the spared nerve injury in anesthetized rats. At 7 days after developing the model, LRRK2 inhibitor GNE-7915 12.5, 25.0 and 50.0 mg/kg were intraperitoneally injected in low-, medium- and high-dose GNE-7915 groups, respectively. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model, at 7 days after developing the model, and at 4 h after injecting the inhibitor. After measurement of the pain threshold, the rats were sacrificed and the spinal cord tissues were taken for determination of the positive expression of ionized calcium-binding adapter molecule 1(Iba-1) (by immunofluorescence staining), contents of interleukin-1β (IL-1β), monocyte chemotactic protein-1 (MCP-1) and IL-18 (by enzyme-linked immunosorbent assay), positive expression of phosphorylated LRRK2 (p-LRRK2) (by immunofluorescence staining), and expression of LRRK2, IL-1β, MCP-1 and IL-18 (by immunoblotting). The ratio of p-LRRK2/LRRK2 was calculated. Results:Compared with C group, the MWT was significantly decreased, the TWL was shortened, the proportion of Iba-1 and p-LRRK2 positive cells in spinal cord tissues, contents of IL-1β, MCP-1 and IL-18, and p-LRRK2/LRRK2 ratio were increased, and the expression of IL-1β, MCP-1 and IL-18 proteins was up-regulated in NP group ( P<0.05). Compared with NP group, the MWT was significantly increased, the TWL was prolonged, the proportion of Iba-1 and p-LRRK2 positive cells in spinal cord tissues, contents of IL-1β, MCP-1 and IL-18, and p-LRRK2/LRRK2 ratio were decreased, and the expression of IL-1β, MCP-1 and IL-18 proteins was down-regulated in low-, medium- and high-dose GNE-7915 groups ( P<0.05). Conclusions:LRRK2 in the spinal cord may be involved in the pathophysiological mechanism of neuropathic pain by activating microglia and inducing inflammatory responses in rats.

Objective: To analyze effects of cooperation between physicians in department of burn surgery and department of intensive care medicine on rescue and treatment of severe mass burn patients involved in August 2nd Kunshan factory aluminum dust explosion accident. Methods: On August 2nd, 2014, 15 extremely severe burn patients involved in August 2nd Kunshan factory aluminum dust explosion accident were admitted to temporary burn treatment center established in Department of Critical Care Medicine of the Second Affiliated Hospital of Soochow University. The 15 patients were equally divided into 3 groups, with 5 patients in each group. Fifteen surgeons and 30 nurses from department of burn surgery and 15 physicians and 30 nurses from department of intensive care medicine from different hospitals in China were divided into 3 groups, with 5 physicians and 10 nurses from department of burn surgery and 5 physicians and 10 nurses from department of intensive care medicine in each group. Each group of physicians and nurses were responsible for treatment of 5 patients. Treatment of patients was leaded by surgeons from department of burn surgery, who were responsible for wound dealing and operation. Physicians from department of intensive care medicine were responsible for systemic treatment and adjustment of relevant equipment's parameters. Volume of fluid infusion and urine output in shock period, severe systemic complication during period of treatment, using time and kind of antibiotics, death in 1 month after admission, length of hospital stay, and survival of patients were monitored. Results: Volume of fluid infusion of 15 extremely severe burn patients within the first 24 hours post injury was 10 360-17 162 (12 998±1 811) mL, including (1.62±0.23) mL·% total body surface area (TBSA)^-1·kg^-1 electrolyte and colloid and (2 850±232) mL glucose, with electrolyte and colloid ratio of (1.76±0.23)∶1.00. Volume of urine output within the first 24 hours post injury was (2 384±1 242) mL, with (99±52) mL in each hour. Volume of fluid infusion of 15 extremely severe burn patients within the second 24 hours post injury was 8 720-11 616 (9 406±1 277) mL, including (1.04±0.22) mL·%TBSA^-1·kg^-1 electrolyte and colloid and (2 910±187) mL glucose, with electrolyte and colloid ratio of (1.53±0.31)∶1.00. Volume of urine output within the second 24 hours post injury of patients was (2 299±1 362) mL , with (108±61) mL in each hour. One patient had pulmonary infection, and 7 patients had fungal infection, and no patient had gut microbiota dysbiosis. Patients were treated with combined 2 kinds of antibiotics for 21-85 (50±16) d. No patient died within 1 month after admission. The length of hospital stay was 53-132 (98±44) d. Ten patients survived finally. Conclusions After being treated by cooperation between physicians in department of burn surgery and department of intensive care medicine, severe mass burn patients involved in August 2nd Kunshan factory aluminum dust explosion accident had hemodynamic stability and could stably experience shock period, with less complication, shorter length of hospital stay, no death within 1 month after admission, more survived patients, which can provide reference for rescue and treatment of severe mass burn patients.