Synergistic protection of matrine and lycopene against lipopolysaccharide­induced acute lung injury in mice.

Acute lung injury (ALI) is a major cause of morbidity and mortality globally, and is characterized by widespread inflammation in the lungs. Increased production of reactive oxygen species is hypothesized to be associated with ALI. Matrine and lycopene are active products present in traditional Chinese medicine. Matrine is an effective inhibitor of inflammation, whereas lycopene decreases lipid peroxidation. Therefore, it was hypothesized that combinatorial treatment with matrine and lycopene may provide synergistic protection against ALI. In the present study, mice were treated with dexamethasone (DEX; 5 mg/kg), matrine (25 mg/kg), lycopene (100 mg/kg), and matrine (25 mg/kg) + lycopene (100 mg/kg) for 7 days prior to injury induction using lipopolysaccharide (LPS; 5 mg/kg) for 6 h. Lung tissues were collected following the sacrifice of the mice and hematoxylin and eosin staining was used for histological analysis. Malondialdehyde (MDA), glutathione (GSH) and myeloperoxidas (MPO) levels were examined by respective kits. The expressions of interleukin­6 (IL­6) and tumor necrosis factor­α (TNF­α) were evaluated by ELISA. The expressions of IκBα and NF­κB p65 were examined by reverse transcription­quantitative polymerase chain reaction, western blotting and immunohistochemistry. The results indicated that the combined treatment exhibited a similar effect to DEX, both of which attenuated lung structural injuries, downregulated the expressions of IL­6, TNF­α, MPO and MDA, and upregulated that of GSH. Furthermore, the combined treatment and DEX inhibited NF­κB p65 activation. The present study revealed that combined treatment with matrine and lycopene exhibited protective effects on an LPS­induced mouse model of ALI, suggesting that they may serve as a potential alternative to glucocorticoid therapy for ALI.

A highly efficient ß-glucosidase from the buffalo rumen fungus Neocallimastix patriciarum W5.

BACKGROUND: Cellulose, which is the most abundant renewable biomass on earth, is a potential bio-resource of alternative energy. The hydrolysis of plant polysaccharides is catalyzed by microbial cellulases, including endo-ß-1,4-glucanases, cellobiohydrolases, cellodextrinases, and ß-glucosidases. Converting cellobiose by ß-glucosidases is the key factor for reducing cellobiose inhibition and enhancing the efficiency of cellulolytic enzymes for cellulosic ethanol production. RESULTS: In this study, a cDNA encoding ß-glucosidase was isolated from the buffalo rumen fungus Neocallimastix patriciarum W5 and is named NpaBGS. It has a length of 2,331 bp with an open reading frame coding for a protein of 776 amino acid residues, corresponding to a theoretical molecular mass of 85.1 kDa and isoelectric point of 4.4. Two GH3 catalytic domains were found at the N and C terminals of NpaBGS by sequence analysis. The cDNA was expressed in Pichia pastoris and after protein purification, the enzyme displayed a specific activity of 34.5 U/mg against cellobiose as the substrate. Enzymatic assays showed that NpaBGS was active on short cello-oligosaccharides from various substrates. A weak activity in carboxymethyl cellulose (CMC) digestion indicated that the enzyme might also have the function of an endoglucanase. The optimal activity was detected at 40°C and pH 5 ~ 6, showing that the enzyme prefers a weak acid condition. Moreover, its activity could be enhanced at 50°C by adding Mg2+ or Mn2+ ions. Interestingly, in simultaneous saccharification and fermentation (SSF) experiments using Saccharomyces cerevisiae BY4741 or Kluyveromyces marxianus KY3 as the fermentation yeast, NpaBGS showed advantages in cell growth, glucose production, and ethanol production over the commercial enzyme Novo 188. Moreover, we showed that the KY3 strain engineered with the NpaNGS gene can utilize 2 % dry napiergrass as the sole carbon source to produce 3.32 mg/ml ethanol when Celluclast 1.5 L was added to the SSF system. CONCLUSION: Our characterizations of the novel ß-glucosidase NpaBGS revealed that it has a preference of weak acidity for optimal yeast fermentation and an optimal temperature of ~40°C. Since NpaBGS performs better than Novo 188 under the living conditions of fermentation yeasts, it has the potential to be a suitable enzyme for SSF.

Fabrication of pre-determined shape of bone segment with collagen-hydroxyapatite scaffold and autogenous platelet-rich plasma.

BACKGROUND: Reconstruction of large segment of bony defects is frequently needed in hand surgery. Autogenous bone grafting is considered the standard in management of these bony defects but has limited source of graft material. Collagen and hydroxyapatite have been used as bone-filling materials and are known to serve as the osteoconductive scaffold for bone regeneration. On the other hand, platelet-rich plasma is a kind of natural source of growth factors, and has been used successfully in bone regeneration and improving wound healing. This study was designed to evaluate the effectiveness of using biohybrids of platelet-rich plasma and collagen-hydroxyapatite beads for fabricating of protrusive bone in a rabbit animal model. METHODS: Biomaterial beads comprised of particulate hydroxyapatite dispersed in fibrous collagen (type I) matrices were prepared and filled in the ringed polytetrafluoroethylene (PTFE) artificial vascular graft (3 cm long, 1 cm in diameter). New Zealand White rabbits were each implanted with two cylindrical PTFE artificial vascular graft over both iliac crests (n = 16). A 2 x 0.5 cm opening was created at the side of each PTFE chamber to allow the content of chamber in contact with the bone marrow of the ileum. The chambers were empty (groups A and D), filled with type I collagen/hydroxyapatite beads (groups B and C). In groups C and D, autologous platelet rich plasma (PRP) was given by transcutaneous injection method into the chambers every week. After 12 weeks, the animals were sacrificed and the chambers were harvested for radiological and histological analysis. RESULTS: In plain radiographs, the group C chambers had significantly higher bone tissue engineered (average calcified density 0.95, average calcified area 61.83%) compared with other groups (P < 0.001). In histological examination, there was a creeping substitution of the implant by the in-growth of fibrovascular tissue in group C. Abundant fibrovascular networks positioned interstitially between these biomaterial beads in all parts of chamber. Degradation of these beads and newly formed capillaries and osteoids around the degraded matrixes are shown. The continually calcification in the matrixes or degraded matrixes is evidenced by the strong green fluorescence observed under the confocal microscope. In group B, looser architecture without evidence of tissue in-growth was shown. In the scaffold absent groups (A and D), there was only fibrous tissue shown within the chamber. CONCLUSIONS: In this study, we have demonstrated a feasible approach to fabricate an osseous tissue that meets clinical need. Using the type I collagen/ hydroxyapatite gel beads matrixes and intermittent injection of autologous platelet-rich-plasma, specific 3D osseous tissues with fibrovascular network structure from pre-exist bony margin were successfully created in an in vivo animal model.

Hydroa vacciniforme-like Epstein-Barr virus-associated monoclonal T-lymphoproliferative disorder in a child.

Hydroa vacciniforme (HV) is a chronic photosensitivity disorder induced by ultraviolet radiation. Hydroa vacciniforme-like lymphoma is a rare cutaneous T-cell lymphoma occurring mainly in childhood. Recent studies have demonstrated an association between chronic latent Epstein-Barr virus (EBV) infection and both the benign skin disorder and the lymphoma. The authors report a 6-year-old boy with chronic EBV infection, HV-like skin eruptions, and chronic hepatitis. Histopathologic examination of a skin biopsy specimen demonstrated epidermal ballooning degeneration and dense superficial and deep perivascular and periappendageal lymphoid cell infiltrates extending to the fat lobules. Some blood vessels in the deep plexus were infiltrated by predominantly CD4+ and TIA-1+ cytotoxic T cells. The EBV genomes were found within tissue from three skin biopsies and peripheral blood cells. Monoclonal T-cell receptor gene rearrangement was present in skin biopsy specimens. Although no lymphoma has been found during 2 years of follow-up treatment, the possibility of lymphoma developing out of the current smoldering stage is of concern. The clinical manifestations of lymphoproliferative disorder and chronic active EBV infection are discussed.