Antitumor peptides from marine organisms.

The biodiversity of the marine environment and the associated chemical diversity constitute a practically unlimited resource of new antitumor agents in the field of the development of marine bioactive substances. In this review, the progress on studies of antitumor peptides from marine sources is provided. The biological properties and mechanisms of action of different marine peptides are described; information about their molecular diversity is also presented. Novel peptides that induce apoptosis signal pathway, affect the tubulin-microtubule equilibrium and inhibit angiogenesis are presented in association with their pharmacological properties. It is intended to provide useful information for further research in the fields of marine antitumor peptides.

Primary salivary gland tumors of the lung: Two cases date report and literature review.

BACKGROUND: Primary salivary gland-type tumors of the lung (PSGTTL) is a rare intrathoracic malignant tumor that accounts for all lungs <1% of tumors. PURPOSE: To introduce two case reports of primary lung salivary gland tumors, and highlight their diagnosis and treatment challenges. CASE REPORT: The first case was a 30-year-old female, who complained of repeated coughing and dyspnea for 1 year and worsening for 2 days. Chest CT and bronchoscopy showed new organisms in the lower part of the trachea, that the bronchus obstruction accounted for 70%. The biopsy histology revealed a adenoid cystic carcinoma. She underwent extensive surgical resection and multiple radiotherapy, and She is recovering well from follow-up. The second case was a 70-year-old man, who complained of intermittent sputum blood for 2 years, worsening hemoptysis and chest tightness for 3 months. The new organisms was found in the upper trachea from Chest CT and bronchoscopy, and histological biopsy was used to diagnose epithelial myoepithelial carcinoma. He underwent twice bronchoscopy thermal ablation treatments. The follow-up is currently in good condition. CONCLUSION: Primary lung salivary gland tumors are considered to be rare malignant tumors in the lungs. Early detection is advocated as late presentation with advanced tumor presents diagnostic and therapeutic challenges.

Controlled release of BMP-2 from titanium with electrodeposition modification enhancing critical size bone formation.

In this study, a porous Ti-alloy based implant with an interconnected channel structure (MAO-CaP-BMP2) is fabricated using a method combining 3D printing, microarc oxidation (MAO) treatment, and co-precipitation of Ca,P layer with BMP-2 technique. The macroporous structure with pore size of 600 µm made by 3D printing not only enhances the ingrowth of cells but also allows the formation of blood vessels inside the implant. As a result, the new bond formation is promoted. In addition, the microporous dioxide layer formed on the implant surface by MAO provides the sites for co-precipitation of Ca,P layer with BMP-2. The microstructure allows the prolonged release of BMP-2. Our results show that a sustained release of BMP-2 over 35 days is achieved for MAO-CaP-BMP2 group longer than Ti without MAO modification group and without Ca,P electrochemical deposition group. The slow release of BMP-2 at the bone/implant interface for a long period of time leads to enhancement of the osseointegration between the implant and surrounding bones. This result indicates that MAO-CaP-BMP2 is a good candidate of growth factor carrier. Successful regeneration of bone requires the concomitant processes of osteogenesis and neovascularization. MAO-CaP-BMP2 modified Ti-alloy implant is both osteoinductive and osteoconductive which can create better osteogenesis and angiogenesis. As a result, it can enhance bone formation.

[Thoracoscopic cardiac surgical procedures: a report of 674 cases].

OBJECTIVE: To evaluate the efficacy and safety of thoracoscopic cardiac surgical procedures under extracorporeal circulation. METHODS: From May 2000 to May 2006, 674 patients received thoracoscopic cardiac surgery under extracorporeal circulation. These procedures included atrial septal defect occlusion for 238 patients, ventricular septal defect occlusion for 380 patients and mitral valve replacement for 56 patients. Thirty degree thoracoscopes and femoral extracorporeal circulation were used. The aorta was cross-clamped and the myocardium was protected by coronary perfusion with cold crystal or blood cardioplegia. RESULTS: The operation succeed in 645 patients (96%, 645/674). Enlarging the incision was performed in 28 patients. Operation time was from 1.8 h to 5.6 h with the mean of (2.8 +/- 1.2) h. Cardiopulmonary bypass time was from 56 min to 198 min with the mean of (78 +/- 2.3) min. Aortic cross-clamp time was from 8 min to 96 min with the mean of (31 +/- 19) min. The volume of chest drainage was (140 +/- 46) ml. None but one postoperative death occurred, the mortality was 0.15%. Postoperative complications occurred in 48 cases (7%), including bleeding in 8 patients, leakage in 5 patients (reoperation in 2 patients) and hemo-pneumothorax in 33 patients. One patient died postoperatively from cerebral hemorrhage (0.15%, 1/647). CONCLUSION: Thoracoscopic cardiac surgical procedures for atrial septal defect occlusion, ventricular septal defect occlusion and mitral valve replacement is feasible and safe.

[Purification and properties of an antimicrobial substance from marine Brevibacillus laterosporus Lh-1].

An antimicrobial substance produced by Brevibacillus laterosporus isolated from the sea sediment was purified and characterized. The antimicrobial substance was purified by ultrafiltration, DEAE-Sepharose Fast flow chromatography, CM-Sepharose Fast flow chromatography and HPLC reversed phase column chromatography, and after the final purification step, one active fraction was obtained, designated R-1. The molecular weight (MW) was accurately determined by MALDI-TOF-MS as 1608.023 Da. And its pI was determined with Rotofor Cell BIO-RAD to be 8.55. Amino acid analysis of the purified R-1 showed that it was composed of Leu, Tyr, Val, Ile, Lys, Gly, Met, Ser and Ala. Most of them were hydrophobic and neutral amino acid except Lys which was a basic amino acid. And this accorded with pI of R-1. R-1 remained active over a wide temperature range and it also was active over a broad pH rang. R-1 was insusceptible to pancreatin, pepsin and alkaline proteinase. Agar radial diffusion assay showed that R-1 had low minimun bactericidal concentration against Gram-Positive Bacteria such as Streptococcus mutans, Staphylococcus aureaus, Clostridium and Gram- egative Bacteria such as Escherichia coli Pseudomonas putrefaciens And R-1 had antibacterial activities against Candida albicans

A polypeptide from Chlamys farreri abolishes UV-induced apoptosis in murine thymocytes in vitro.

Previously we reported that a polypeptide from Chlamys farreri (PCF) was a potent photoprotective agent against ultraviolet (UV) irradiation in vitro. To understand the mechanism by which PCF protects cells from irradiation, we studied anti-apoptotic effects of PCF against UV irradiation on the murine thymocytes in vitro. MTT and flow cytometric analysis assays showed that 2h pretreatment with PCF completely abolished UV induced cell death. TEM examination showed that PCF fully protected the ultrastructure of thymocytes exposed to UV irradiation. Lipid peroxidation and intracellular reactive oxygen species assays indicated that PCF efficiently blocked production of reactive oxygen intermediates induced by UV irradiation. Further, PCF protected UV-irradiated thymocytes from losing mitochondrial transmembrane potential and DNA fragmentation. Based on these observations we propose that PCF is a potent anti-apoptotic factor, which protects cells from irradiation at multiple steps.

Polypeptide from Chlamys farreri inhibits murine thymocytes apoptosis and modulates UVB induced signaling pathway activation.

Polypeptide from Chlamys farreri (PCF) has been identified as a potent antioxidant and photoprotective agent. In this study, we investigated whether PCF could inhibit apoptosis of murine thymocytes induced by ultraviolet B (UVB) and modulate UVB induced the mitogen-activated protein kinases (MAPKs) cascade in vitro. Our results show that PCF inhibit UVB-induced apoptotic cell death in murine thymocytes. We also found that PCF potently stimulated the phosphorylation of ERKs, which is involved in the cell survival-signaling cascade. Furthermore, the specific inhibition of the ERKs pathways by PD98059 reduced the cytoprotective effect of PCF. On the other hand, the JNKs and p38 inhibitor SP600125 and SB203580 additively enhanced the cytoprotective effect of PCF. We concluded that the activation of JNKs and p38 kinase played an important role in UVB-induced apoptosis, and PCF likely exerted its cytoprotective effect in thymocytes through ERKs activation. These suggested that part of the antiapoptotic effect of PCF might be mediated by its ability to modulate the MAPKs cascade.

Polypeptide from Chlamys farreri protects HaCaT cells from UVB-induced apoptosis.

A natural polypeptide from marine Chlamys farreri (a kind of scallop) (PCF), has been recently been found to be an effective photoprotective agent against ultraviolet rays B (UVB)-induced mitochondria damage in normal human fibroblasts. To investigate whether PCF has the antiapoptotic effect on human keratinocytes, in the present study, we established an apoptotic model on HaCaT cell line by means of UVB radiance of 30 mJ/cm(2) and compared the effect of different PCF treatments on UVB-radiated cells. Flow cytometry analyses showed that PCF treatment before UVB-irradiation inhibited UVB-induced apoptosis, the loss of mitochondrial membrane potential (Deltapsim) and the increase of free Ca(2+) level in HaCaT cells. In parallel with these results, UVB-irradiation enhanced activities of caspases-3, 8, 9, while this enhancement was inhibited by PCF treatment prior to irradiation. PCF added after irradiation neither reduced UVB-induced activities of the three caspases nor synergized the effect of pre-added PCF. Cellular ultrastructural features obtained from transmission electron microscopy further confirmed the antiapoptotic effect of PCF pre-treatment. It is concluded that the antiapoptotic effect of PCF is not therapeutic but prophylactic. Caspases-3, 8, 9, Deltapsim and calcium are involved in UVB-induced apoptosis, while prophylactic PCF inhibits apoptosis of UVB-irradiated HaCaT cells by blocking the caspases activities, the Deltapsim lost and the elevation of intracellular free Ca(2+) level.

[Study on low-temperature alkaline lipase from Bohaisea-9145].

Lipase-overproducing marine yeast Bohaisea-9145 isolated from the Bohai sea region was identified as psychrotrophlic Yarrowia lipolytica. The optimum composition of culture medium was ground soybean 4%, rice bran 4%, peanut power 4%, crude peanut oil 0.5%, MgSO4 0.05%, KH2PPO4 0.2%. The optimum temperature and pH were 26 +/- 1 degrees C pH5.0, the time of fermentation cycle was 23h. The lipase activity was improved to 258.67 U/mL through optimization, four-fold more than that of initial strain. The lipase displayed maximum activity at pH 8.5, 35 degrees C and was stable within the range of pH 4.0 - 9.0 at low temperature. It had high thermosensitivity, good compatibility with familiar metal ions and chemical reagents. It was antioxidant and had strong resistance to high salinity. As a novel marine low-temperature lipase, it has promising prospect in many fields, especially as an additive of detergent.

Polypeptide from Chlamys farreri attenuates murine thymocytes damage induced by ultraviolet B.

AIM: Polypeptide from Chlamys farreri (PCF, molecular mass is 879) is a new marine polypeptide compound isolated from Chlamys farreri. This study investigates the possible protective roles and the mechanism of PCF against ultraviolet B (UVB)-induced apoptosis in murine thymocytes. METHODS: The rate of apoptosis and caspase-3 activation was measured by flow cytometry. The expression of stress-response genes c-fos and c-jun was observed by RT-PCR. Western blot analysis was performed to determine the release of cytochrome c. RESULTS: It was found that UVB induced murine thymocyte death. The cells treated with UVB showed an increase in cytochrome c release, caspase-3 activity, as well as in the expression of c-fos and c-jun. In addition, all were involved in UVB-induced cell apoptosis. CONCLUSION: Our present observations pointed to the ability of PCF to avert UVB-induced apoptosis in thymocytes by modulating c-fos and c-jun expression, cytochrome c release, and the consequent activation of caspase-3, which were essential components of the UV-induced cell apoptotic pathway. The results suggested that PCF is a promising protective substance against UV radiation.

[Purification and characterization of a lysozyme from a marine microorganism].

A novel lysozyme was purified from a marine microorganism and its major characteristics were studied. Cell-free supernatant was prepared by centrifugation of culture broth, ultrafiltration using a hollow fiber (molecular weight cut off, 50kD) and concentration using a hollow fiber (molecular weight cut off, 10kD). The crude lysozyme was purified 34.7 fold to electrophoretic homogeneity with a recovery of 24.1% by CM-Sepharose FF cationic-exchange and Sephadex G-100 gel chromatography. The relative molecular weight of this lysozyme was determined as about 39 kD. The optimum pH and temperature towards Micrococcus lysodleikticus were pH 8.0 and 35 degrees C respectively, and the enzyme was stable at temperature below 50 degrees C and pH 5.0 - 10.0. The lysozyme activity was slightly enhanced by Zn2+ and Cu2+ and slightly inhibited by Mn2+ and Ag+. The lysozyme showed good compatibility to many common chemical agents such as EDTA (0.1%) and KH2 PO4 (1.0%). The lysozyme had broad-spectrum against many bacteria, including a number of pathogens, which were resistant to egg-white lysozyme.

Inhibitory effect of polypeptide from Chlamys farreri on ultraviolet A-induced oxidative damage on human skin fibroblasts in vitro.

We have previously reported that polypeptide from Chlamys farreri (PCF) inhibits the oxidative damage of ultraviolet A (UVA) on HeLa cells in vitro [Acta Pharm. Sin. 23 (2002) 961]. To further elucidate a possible role for PCF on UVA-damaged normal human cells, we established the oxidative damage models of normal human dermal fibroblasts (NHDF) exposed to UVA to study the protective effect of PCF on human dermal fibroblasts in vitro. In this study, 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) method was used to detect the cell viability. The intracellular superoxide dismutase (SOD), glutathione peroxidase (GSH-px), catalase (CAT), xanthine oxidase (XOD), malondialdehyde (MDA), reactive oxygen species (ROS), total antioxidative capacity (T-AOC), and anti-superoxide anion capacity (A-ASC) were measured. The effect of PCF on UVA-induced apoptosis were investigated by Annexin V-FITC assay. Intracellular calcium was determined with the calcium-sensitive fluorochrome Fluo-3, and mitochondrial transmembrane potential with rhodamine 123. Comet assay was employed to detect the UVA-induced DNA damage. The ultrastructure of cell was observed under transmission electron microscope. The results indicated that PCF could greatly enhance the viability of NHDF and markedly promote SOD, GSH-px, T-AOC, and A-ASC, while the amounts of MDA and ROS, the activity of XOD were decreased. PCF could inhibit UVA-induced apoptosis and DNA damage in NHDF. The concentration of cellular free calcium was decreased and the mitochondrial transmembrane potential was increased by PCF. In ultrastructure of NHDF, PCF could greatly decrease UVA-induced damage, especially membrane. Our results suggest that the supplementation of PCF appears to reduce the UVA-induced normal human dermal fibroblasts damage efficiently. It may be involved in the PCF's abilities of scavenging oxygen free radical, inhibiting lipid peroxidation, increasing antioxidative enzymes, decreasing intracellular calcium and protection of membrane structure in NHDF irradiated by UVA.

Protective effect of polypeptide from Chlamys farreri on hairless mice damaged by ultraviolet A.

AIM: To study the protective effect of the polypeptide isolated from Chlamys farreri (PCF) on hairless mice skin damaged by ultraviolet A. METHODS: Enzymes and malondialdehyde (MDA) were determined by biochemical methods; the expressions of Bcl-2 protein and NOS protein were examined by immunohistochemical technique. The ultra-structure of the skin was observed through electronic microscope. RESULTS: PCF could enhance the activities of glutathione peroxidase (GSH-px), superoxide dismutase (SOD), and total anti-oxidative capacity (T-AOC). Also PCF could reduce the amount of MDA, increase the expression of Bcl-2 protein, and inhibit the expression of NOS protein. The ultra-structure of epidermis and fibroblasts remained normal in 20 % PCF groups; there were vacuoles in smooth endoplasm reticulum in epidermis of mice and the number of rough endoplasm reticulum in fibroblasts was decreased in model group. CONCLUSION: PCF had the protective effects on hairless mice skin damaged by ultraviolet A via its anti-oxidative mechanisms.

Protective effect of polypeptide from Chlamys farreri on mitochondria in human dermal fibroblasts irradiated by ultraviolet B.

AIM: To study the effect of polypeptide from Chlamys farreri (PCF) on mitochondria of human dermal fibroblasts irradiated by ultraviolet B (UVB) in vitro. METHODS: Malondialdehyde (MDA) and antioxidant enzymes including superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) were determined by biochemical methods. Mitochondrial transmembrane potential was measured by flow cytometry. Ultrastructure of fibroblasts was observed with transmission electron microscope. RESULTS: UVB (1.176 x 10(-4) J/cm(2)) induced mitochondria damage in dermal fibroblast and PCF (0.25%-1%) reduced the damage in a concentration-dependent manner. Furthermore, PCF also concentration-dependently maintained the stability of mitochondrial transmembrane potential. PCF was able to reduce the MDA formation caused by UVB, meanwhile increased the activities of SOD and GSH-PX. The differences among the PCF groups and UVB model group were significant (P<0.05, P<0.01). CONCLUSION: The UVB-induced mitochondria damage was alleviated by PCF in human dermal fibroblasts.