Mesenchymal stem cell (MSC)-derived exosomes (Exos) were reported to a prospective candidate in accelerating diabetic wound healing due to their pro-angiogenic effect. MSCs pretreated with chemistry or biology factors were reported to advance the biological activities of MSC-derived exosomes. Hence, this study was designed to explore whether exosomes derived from human umbilical cord MSCs (hucMSCs) preconditioned with Nocardia rubra cell wall skeleton (Nr-CWS) exhibited superior proangiogenic effect on diabetic wound repair and its underlying molecular mechanisms. The results showed that Nr-CWS-Exos facilitated the proliferation, migration and tube formation of endothelial cells in vitro. In vivo, Nr-CWS-Exos exerted great effect on advancing wound healing by facilitating the angiogenesis of wound tissues compared with Exos. Furthermore, the expression of circIARS1 increased after HUVECs were treated with Nr-CWS-Exos. CircIARS1 promoted the pro-angiogenic effects of Nr-CWS-Exos on endothelial cellsvia the miR-4782-5p/VEGFA axis. Taken together, those data reveal that exosomes derived from Nr-CWS-pretreated MSCs might serve as an underlying strategy for diabetic wound treatment through advancing the biological function of endothelial cells via the circIARS1/miR-4782-5p/VEGFA axis.
Humans ; Endothelial Cells/metabolism* ; Exosomes/metabolism* ; Cell Wall Skeleton/metabolism* ; Neovascularization, Physiologic ; Wound Healing/physiology* ; MicroRNAs/metabolism* ; Diabetes Mellitus ; Vascular Endothelial Growth Factor A/metabolism*

Peach brown rot, caused by Monilinia fructicola, is one of the most serious peach diseases. A strain belonging to the Actinomycetales, named Streptomyces blastmyceticus JZB130180, was found to have a strong inhibitory effect on M. fructicola in confrontation culture. Following the inoculation of peaches in vitro, it was revealed that the fermentation broth of S. blastmyceticus JZB130180 had a significant inhibitory effect on disease development by M. fructicola. The fermentation broth of S. blastmyceticus JZB130180 had an EC₅₀ (concentration for 50% of maximal effect) of 38.3 µg/mL against M. fructicola, as determined in an indoor toxicity test. Analysis of the physicochemical properties of the fermentation broth revealed that it was tolerant of acid and alkaline conditions, temperature, and ultraviolet radiation. In addition, chitinase, cellulase, and protease were also found to be secreted by the strain. The results of this study suggest that S. blastmyceticus JZB130180 may be used for the biocontrol of peach brown rot.
Ascomycota/pathogenicity* ; Bacterial Proteins/metabolism* ; Cell Wall/metabolism* ; Cellulase/metabolism* ; Chitinases/metabolism* ; Fermentation ; Fruit/microbiology* ; Pest Control, Biological/methods* ; Phylogeny ; Plant Diseases/prevention & control* ; Prunus persica/microbiology* ; Siderophores/metabolism* ; Streptomyces/physiology*

OBJECTIVE: To study the effects of aerobic exercise combined with chlorella pyrenoidos of disintegrated cell wall on the lipid metabolism in rats with high-fat diet. METHODS: Fifty-five male Wistar rats were subjected to adaptive feeding for 4 days and weight-free swimming training for 3 days, 20 min/d. After eliminating 5 rats that were not suitable for swimming training, the other rats were randomly divided into 5 groups according to their body weight:control group (C group), high fat diet group (H group), high-fat diet + chlorella group(HC group), high fat diet + aerobic exercise group (HM group), high fat diet + chlorella + aerobic exercise group (HMC group), 10 in each group. The HM and HMC group were subjected to 60 min/d swimming training for 6 weeks with non-weight-bearing. Group C were fed regular diet. The other groups were fed with high-fat diet, the rats in group HC and HMC were intragastrically treated with chlorella pyrenoidos of disintegrated cell wall at the dose of 3.9 g/(kg·d), the volume was 5 ml/kg, and the other groups are given equivalent saline. The Lee's index and biochemical indexes of blood and liver were measured after 6 weeks. RESULTS: Compared with group C, Lee's index, serum levels of free fatty acids(FFA), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-c), liver FFA and interleukin-10 (IL-10) were increased significantly (<0.01), the serum level of high-density lipoprotein cholesterol (HDL-c) was decreased significantly (<0.01) in group H. Compared with group H, Lee's index, serum FFA, IL-6, TNF-α, TC, TG, LDL-c, liver FFA and IL-10 were decreased significantly (<0.05 or <0.01), serum level of HDL-c was increased significantly (<0.05 or <0.01) in group HC, HM and HMC. Compared with group HC and HM, Lee's index, serum FFA, IL-6, TNF-α, TC, TG, LDL-c, liver FFA and IL-10 were decreased significantly (<0.05), serum level of HDL-c was increased significantly (<0.05) in group HMC. CONCLUSIONS Aerobic exercise and chlorella pyrenoidos of disintegrated cell wall can improve lipid metabolism in rats with high-fat diet and reduce the lipid toxicity caused by obesity. Joint intervention is more effective than single intervention.
Animals ; Cell Wall ; Chlorella ; Diet, High-Fat ; Lipid Metabolism ; Male ; Physical Conditioning, Animal ; Rats ; Rats, Wistar

Small, cysteine-rich, highly stable antifungal proteins secreted by filamentous Ascomycetes have great potential for the development of novel antifungal strategies. However, their practical application is still limited due to their not fully clarified mode of action. The aim of this work was to provide a deep insight into the antifungal mechanism of Neosartorya fischeri antifungal protein (NFAP), a novel representative of this protein group. Within a short exposure time to NFAP, reduced cellular metabolism, apoptosis induction, changes in the actin distribution and chitin deposition at the hyphal tip were observed in NFAP-sensitive Aspergillus nidulans. NFAP did show neither a direct membrane disrupting-effect nor uptake by endocytosis. Investigation of A. nidulans signalling mutants revealed that NFAP activates the cAMP/protein kinase A pathway via G-protein signalling which leads to apoptosis and inhibition of polar growth. In contrast, NFAP does not have any influence on the cell wall integrity pathway, but an unknown cell wall integrity pathway-independent mitogen activated protein kinase A-activated target is assumed to be involved in the cell death induction. Taken together, it was concluded that NFAP shows similarities, but also differences in its mode of antifungal action compared to two most investigated NFAP-related proteins from Aspergillus giganteus and Penicillium chrysogenum.
Actins ; metabolism ; Antifungal Agents ; pharmacology ; Apoptosis ; drug effects ; Aspergillus nidulans ; cytology ; drug effects ; growth & development ; Cell Membrane ; drug effects ; metabolism ; Cell Wall ; drug effects ; metabolism ; Chitin ; metabolism ; Endocytosis ; drug effects ; Fungal Proteins ; pharmacology ; GTP-Binding Proteins ; metabolism ; Hyphae ; cytology ; drug effects ; Microbial Viability ; drug effects ; Neosartorya ; chemistry ; Signal Transduction ; drug effects

Acanthamoeba cysts are resistant to unfavorable physiological conditions and various disinfectants. Acanthamoeba cysts have 2 walls containing various sugar moieties, and in particular, one third of the inner wall is composed of cellulose. In this study, it has been shown that down-regulation of cellulose synthase by small interfering RNA (siRNA) significantly inhibits the formation of mature Acanthamoeba castellanii cysts. Calcofluor white staining and transmission electron microscopy revealed that siRNA transfected amoeba failed to form an inner wall during encystation and thus are likely to be more vulnerable. In addition, the expression of xylose isomerase, which is involved in cyst wall formation, was not altered in cellulose synthase down-regulated amoeba, indicating that cellulose synthase is a crucial factor for inner wall formation by Acanthamoeba during encystation.
Acanthamoeba castellanii/*enzymology/genetics/metabolism ; Aldose-Ketose Isomerases/*biosynthesis ; Amebiasis/*pathology ; Benzenesulfonates ; Cell Wall/chemistry/genetics/*metabolism ; Cellulose/biosynthesis ; Down-Regulation ; Encephalitis/parasitology ; Glucosyltransferases/*biosynthesis/genetics ; Keratitis/parasitology ; Microscopy, Electron, Transmission ; RNA Interference ; RNA, Small Interfering

Adult ; CD56 Antigen ; metabolism ; Diagnosis, Differential ; Granzymes ; metabolism ; Humans ; Lymphoma, Extranodal NK-T-Cell ; metabolism ; pathology ; surgery ; Lymphoma, T-Cell, Peripheral ; metabolism ; pathology ; Male ; Pectoralis Muscles ; pathology ; surgery ; RNA, Viral ; metabolism ; Soft Tissue Neoplasms ; metabolism ; pathology ; surgery ; Thoracic Wall ; pathology ; surgery

To improve the extraction of solanesol from tobacco waste, we developed an enzymatic cell wall-breaking process with combined cellulase and ligninase. The effects of reaction time, temperature, pH and enzyme/substrate ratio were determined. The results show that the catalytic effect was better than either single enzyme when the ratio of cellulase to ligninase was 15:1 (U/U). Under the optimized conditions of 175 U/g (enzymes/substrate), tobacco to water 1:5 (W/W), temperature 40 degrees C and pH 6.0, the concentration of solanesol in the solution could reach 0.33 g/L after 8 h. And the average leaching rate reached 96.53% which was 1.68 times of the extraction methods of chemical reflux. It provides new way for the extraction of solanesol from tobacco waste, and worthwhile to be further explored.
Cell Wall ; metabolism ; Cellulase ; metabolism ; Oxygenases ; metabolism ; Plant Leaves ; chemistry ; Terpenes ; isolation & purification ; Tobacco ; chemistry

The mature cyst of Acanthamoeba is highly resistant to various antibiotics and therapeutic agents. Cyst wall of Acanthamoeba are composed of cellulose, acid-resistant proteins, lipids, and unidentified materials. Because cellulose is one of the primary components of the inner cyst wall, cellulose synthesis is essential to the process of cyst formation in Acanthamoeba. In this study, we hypothesized the key and short-step process in synthesis of cellulose from glycogen in encysting Acanthamoeba castellanii, and confirmed it by comparing the expression pattern of enzymes involving glycogenolysis and cellulose synthesis. The genes of 3 enzymes, glycogen phosphorylase, UDP-glucose pyrophosphorylase, and cellulose synthase, which are involved in the cellulose synthesis, were expressed high at the 1st and 2nd day of encystation. However, the phosphoglucomutase that facilitates the interconversion of glucose 1-phosphate and glucose 6-phosphate expressed low during encystation. This report identified the short-cut pathway of cellulose synthesis required for construction of the cyst wall during the encystation process in Acanthamoeba. This study provides important information to understand cyst wall formation in encysting Acanthamoeba.
Acanthamoeba castellanii/*enzymology/genetics/growth & development ; Amebiasis/*parasitology ; Cell Wall/*metabolism ; Cellulose/*biosynthesis/genetics ; Glucosyltransferases/genetics/metabolism ; Glycogen Phosphorylase/genetics/metabolism ; Protozoan Proteins/genetics/*metabolism ; UTP-Glucose-1-Phosphate Uridylyltransferase/genetics/metabolism

OBJECTIVETo establish a human gallbladder carcinoma cell line derived from a metastatic gallbladder carcinoma and identify its biological characteristics.

METHODSTissue samples were separated from the surgical specimen obtained from a patient with metastatic carcinoma and single-cell suspension was prepared. Then the cells were cultured in DMEM medium supplemented with 15% fetal bovine serum. The morphology of tumor cells was observed under an electron microscope. The cell growth curve was plotted. The tumorigenicity of the cell line was studied by subcutaneous inoculation in SCID mice. The cells were infected by lentiviral vector carrying fluorescent report genes (lenti-GFP and lenti-Red2) separately for expressions of GFP and Red2, respectively.

RESULTSA novel metastatic gallbladder carcinoma cell line was successfully established and named "EH-GB1". It could be passaged for over 20 generations with typical malignant epithelial morphology and a stable growth cycle of 24 h. Tumors were formed in all of the 10 SCID mice inoculated with EH-GB1 cells subcutaneously, and the tumor cells were tumor marker CA19-9-positive. Continuous expressions of fluorescent report genes were observed in EH-GB1 cells infected by lenti-GFP and lenti-Red2.

CONCLUSIONEH-GB1 cells might be the first stable cell line of human gallbladder carcinoma established from a metastatic focus of gallbladder carcinoma. This cell line with continuous expressions of GFP and Red2 might be a novel and perfect experimental model for clinical and basic research on gallbladder carcinoma.

Abdominal Neoplasms ; metabolism ; pathology ; secondary ; Abdominal Wall ; Adenocarcinoma ; metabolism ; pathology ; secondary ; Animals ; CA-19-9 Antigen ; metabolism ; Cell Line, Tumor ; metabolism ; pathology ; Female ; Gallbladder Neoplasms ; metabolism ; pathology ; Genes, Reporter ; Green Fluorescent Proteins ; metabolism ; Humans ; Mice ; Mice, Nude ; Mice, SCID ; Middle Aged ; Neoplasm Transplantation

BACKGROUNDPsoriasis is a common inflammatory skin disease, yet knowledge of the factors that may induce, trigger, or exacerbate psoriasis is not fully delineated. Recent advances have improved our understanding of the link between psoriasis and cell-wall-deficient bacteria (CWDB) infections. In the present study we assessed the prevalence of CWDB infection in patients with psoriasis.

METHODSThe carriage rate of CWDB in the tonsil or pharynx of psoriasis patients, chronic tonsillitis patients and controls were investigated using hypertonic medium. Psoriasis patients with CWDB were randomly assigned to two groups and respectively treated with antibiotics or systemic therapy without antibiotic. Human peripheral blood mononuclear cells (PBMC) from psoriasis patients, chronic tonsillitis patients and control subjects were stimulated with bacteria antigens and extra-cellular levels of interferon-gamma (IFN-gamma) and interleukin (IL)-10 were measured in the supernatants using the ELISA technique, in vitro. Meanwhile, the proliferation ability of PBMC to respond to bacteria antigens was detected by MTT assay.

RESULTSCWDB were isolated from 74.2% of psoriasis patients, 23.5% of chronic tonsillitis patients and only 6.3% of controls. Antibiotic therapy was appropriate for approximately 80% of psoriasis patients with CWDB infection, and in only 8.9% psoriasis patients CWDB infection was detected after antibiotic therapy. Meanwhile, our study showed that CWDB and wide-type bacteria did remarkably enhance the production of IFN-gamma, in vitro, and PBMC proliferation.

CONCLUSIONCWDB infection may be a virtual triggering factor in psoriasis by regulating T-cell activation.

Adult ; Anti-Bacterial Agents ; therapeutic use ; Bacteria ; cytology ; drug effects ; isolation & purification ; Cell Wall ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Male ; Middle Aged ; Psoriasis ; drug therapy ; etiology ; metabolism ; microbiology ; Young Adult