On the base of element and metabolism balancing,the mathematic model of the human-like collagen expression phase with recombinant Escherichia coli BL21 was developed and the unknown parameters in the model were estimated with the method of nonlinear optimization.The model was in agreement with the growth kinetics and the metabolic kinetics,and the key calculated parameters of ?h,?p and mx were 1.173 mol?C-mol-1,293.814 mol?C-mol-1 and 17.878 mol?C-mol-1?h-1 respectively.This model could preferably predict the macroscopic reaction rates,and in the synthesis phase of human-like collagen,the specific growth rate should be controlled at 0.04 h-1 with controlling glucose feeding rate to gain the highest specific production rate of human-like collagen.

Diabetic ulcer is recognized as a chronic nonhealing wound, often associated with bacterial infection and tissue necrosis, which seriously affect patients' health and quality of life. The traditional treatment methods exist some problems, such as bacterial resistance and secondary trauma, so it is urgent to find new methods to meet the requirements of diabetic ulcer treatment. In this study, we prepared a drug delivery system (DFO@CuS nanoparticles) based on hollow copper sulfide (CuS) nanoparticles loaded with deferoxamine (DFO), which realized the synergistic therapy of promoting angiogenesis and photothermal antibacterial. The morphological structure and particle size distribution of DFO@CuS nanoparticles were characterized by transmission electron microscopy and particle size analyzer, respectively. The antibacterial effect of DFO@CuS nanoparticles was evaluated by the plate coating method. The effects of DFO@CuS nanoparticles on the proliferation, migration, and tube formation of human umbilical vein endothelial cells (HUVECs) were evaluated by CCK-8 (cell counting kit-8) assay, cell scratch assay, and tube formation assay. The results showed that DFO@CuS nanoparticles were hollow and spherical in shape with an average particle size of (200.9 ± 8.6) nm. DFO@CuS nanoparticles could effectively inhibit the growth of methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa (PA) under near-infrared (NIR) light irradiation. DFO@CuS nanoparticles showed negligible cytotoxicity and effective acceleration of cell migration and tube formation in a certain concentration range. In conclusion, the prepared DFO@CuS nanoparticles exhibit good photothermal antibacterial properties and pro-angiogenic effects, providing a basis for their application in the treatment of diabetic ulcer.

Background and Objective: The expression of transcription factor Elf-1 and inhibitor of apoptosis survivin in non-small cell lung cancer(NSCLC)is correlated with the angiogenic factor vascular endothelial growth factor(VEGF),and are both factors affecting the cell cycle.This study investigated the expression of Elf-1,survivin,and intratumoral microvessel density(iMVD)assessed by monoclonal antibody CD105 in NSCLC,and explored their correlations with clinicopathologic features and angiogenesis of NSCLC.Methods: PowerVisionTM-9000 immunohistochemistry was used to evaluate the expression of Elf-1,survivin,and CD105 in tissue microarrays containing60 specimens of NSCLC and 9 specimens of normal tissue.Western blot analysis was used to evaluate the protein levels of Elf-1 and survivin in 17specimens of NSCLC and 5 specimens of normal tissue.Results: Elf-1 and survivin were detected in 1 of the 9 normal tissues.The positive rates of Elf-1and survivin in NSCLC were 70.0% and 65.0%,respectively.The expression levels of both Elf-1 and survivin were significantly related to tumor differentiation,lymphatic metastasis,clinical stage,and postoperative survival time(P<0.05).Overexpression of both were related to poor prognosis: the survival rates were significantly lower in patients with positive expression than in those with negative expression(P<0.01).Elf-1 expression was positively correlated with survivin expression(r=0.769,P<0.01.Elf-1 and survivin expressions were positively correlated with iMVD(r=0.446,P<0.01 ; r=0.435,P<0.01).Conclusions: The expression of Elf-1 and survivin in NSCLC is related to differentiation,lymphatic metastasis,clinical stage,and prognosis,and both are positively correlated with iMVD.Detection their combined expression can help to predict the malignant behavior of NSCLC.Blocking the activity of Elf-1 and survivin may be a new way to inhibit angiogenesis in NSCLC.

Abdominal Pain ; etiology ; Acute Pain ; etiology ; Child ; Humans ; Lymphoma ; complications

Cranial Fossa, Anterior ; Humans ; Liposarcoma, Myxoid ; diagnostic imaging ; metabolism ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; S100 Proteins ; metabolism ; Skull Base Neoplasms ; diagnostic imaging ; metabolism ; pathology ; Tomography, X-Ray Computed

BACKGROUND AND OBJECTIVEThe expression of transcription factor Elf-1 and inhibitor of apoptosis survivin in non-small cell lung cancer (NSCLC) is correlated with the angiogenic factor vascular endothelial growth factor (VEGF), and are both factors affecting the cell cycle. This study investigated the expression of Elf-1, survivin, and intratumoral microvessel density (iMVD) assessed by monoclonal antibody CD105 in NSCLC, and explored their correlations with clinicopathologic features and angiogenesis of NSCLC.

METHODSPowerVision(TM)-9000 immunohistochemistry was used to evaluate the expression of Elf-1, survivin, and CD105 in tissue microarrays containing 60 specimens of NSCLC and 9 specimens of normal tissue. Western blot analysis was used to evaluate the protein levels of Elf-1 and survivin in 17 specimens of NSCLC and 5 specimens of normal tissue.

RESULTSElf-1 and survivin were detected in 1 of the 9 normal tissues. The positive rates of Elf-1 and survivin in NSCLC were 70.0% and 65.0%, respectively. The expression levels of both Elf-1 and survivin were significantly related to tumor differentiation, lymphatic metastasis, clinical stage, and postoperative survival time (P < 0.05). Overexpression of both were related to poor prognosis: the survival rates were significantly lower in patients with positive expression than in those with negative expression (P < 0.01). Elf-1 expression was positively correlated with survivin expression (r = 0.769, P < 0.01). Elf-1 and survivin expressions were positively correlated with iMVD (r = 0.446, P < 0.01; r = 0.435, P < 0.01).

CONCLUSIONSThe expression of Elf-1 and survivin in NSCLC is related to differentiation, lymphatic metastasis, clinical stage, and prognosis, and both are positively correlated with iMVD. Detection their combined expression can help to predict the malignant behavior of NSCLC. Blocking the activity of Elf-1 and survivin may be a new way to inhibit angiogenesis in NSCLC.

Adult ; Aged ; Antigens, CD ; metabolism ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cell Differentiation ; Endoglin ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Microvessels ; metabolism ; Middle Aged ; Neoplasm Staging ; Neovascularization, Pathologic ; metabolism ; pathology ; Nuclear Proteins ; metabolism ; Receptors, Cell Surface ; metabolism ; Survival Rate ; Transcription Factors ; metabolism

The effects of hypoxia on the level of reactive oxygen species (ROS), IkappaBalpha tyrosine phosphorylation, transcription of P65 mRNA and NF-kappaB activation in isolated rat peritoneal macrophages were investigated by DCFH-DA fluorescence spectrophotometry, Western blotting and RT-PCR. The results obtained are as follows. (1) During hypoxia, the levels of intracellular ROS began to increase at 1 h, then reached a peak at 2 h, and began to decrease after 3 h. IkappaBalpha tyrosine phosphorylation began to rise after 2 h hypoxia and was the highest after 3 h hypoxia. After 4 h hypoxia it decreased gradually. NF-kappaB activation began to increase after 3 h hypoxia, and reached a peak after 4 h hypoxia. (2) When antioxidant NAC (500 mmol/L) was added into the medium, the level of IkappaBalpha phosphorylation showed no significant changes during hypoxia. After adding protein tyrosine kinase inhibitor genistein (200 micromol/L), NF-kappaB activation induced by hypoxia was blocked significantly. (3) The expression of p65 mRNA was also elevated markedly during hypoxia. These results suggest that hypoxia may lead to IkappaBalpha phosphorylation and NF-kappaB activation through intracellular ROS, and that the regulation of NF-kappaB activity may involve IkappaBalpha phosphorylation and the expressions of each subunit gene of NF-kappaB.
Animals ; Cell Hypoxia ; Cells, Cultured ; Macrophages, Peritoneal ; cytology ; physiology ; Mice ; NF-kappa B ; biosynthesis ; genetics ; physiology ; Phosphorylation ; RNA, Messenger ; biosynthesis ; genetics ; Reactive Oxygen Species ; analysis ; Signal Transduction

OBJECTIVETo study the therapeutic effect of intranasal administration of temozolomide (TMZ) for brain-targeting delivery in a rat model bearing orthotopic C6 glioma xenografts.

METHODSForty Wistar rat bearing brain C6 glioma xenograft were randomly divided into 4 groups and treated with physiological saline solution or with TMZ by intravenous injection, gavage or intranasal administration. The tumor size, rat survival time and pathological changes were observed in each group.

RESULTSMagnetic resonance imaging showed a significantly reduced volume of glioma in intranasal TMZ group compared with that in the control, intraveneous TMZ injection group and TMZ gavage groups (12.45∓2.49 mm(3) vs 60.16∓4.12, 33.17∓3.56, and 35.16∓4.36 mm(3), respectively, P<0.05). The median survival time of the C6 glioma-bearing rats was also significantly longer in intranasal TMZ group than in the other 3 groups (31.0 days vs 20, 19, and 21.5 days, respectively, P<0.05). In the glioma xenografts, PCNA expression was the lowest and tumor cell apoptosis rate the highest in intranasal TMZ group.

CONCLUSIONIntranasal TMZ administration can suppress the growth of C6 glioma in rats and may serve as an effective strategy for glioma treatment.

Administration, Intranasal ; Animals ; Antineoplastic Agents, Alkylating ; administration & dosage ; Apoptosis ; Brain Neoplasms ; drug therapy ; Cell Line, Tumor ; Dacarbazine ; administration & dosage ; analogs & derivatives ; Drug Delivery Systems ; Glioma ; drug therapy ; Magnetic Resonance Imaging ; Neoplasm Transplantation ; Rats ; Rats, Wistar

OBJECTIVEDendritic cell vaccines are one of the important active immunotherapies for neoplasms. The aim of this study was to observe the killing effect of specific cytotoxic T lymphocytes (CTL) on liver carcinoma HepG2 and SMMC-7721 cells in vitro. The CTL was induced by human peripheral blood mononuclear cells-originated dendritic cells (DC) transfected by recombinant adeno-associated virus (rAAV) with hAFP gene fragment (137-145).

METHODSImmature DCs were generated from peripheral blood mononuclear cells of healthy volunteers and then transfected by rAAV with AFP gene fragment. The CTL was thereafter induced. The activities of DC and CTL were measured and the killing effect of the CTL on HepG2 cells was detected using M1Tr assay.

RESULTSThe mature DC, transfected or not, highly expressed CD40, CD86 and IL-12. IFN-gamma was highly expressed in the CTL. The DC-induced CTL could effectively recognize and destroy the HepG2 and SMMC-7721 cells.

CONCLUSIONDC transfected by rAAV can stimulate the proliferation and differentiation of lymphocytes and also induce the proliferation of CTL, and their own phenotypes are not significantly altered. The DC vaccine can be effectively used as an adjuvant immunotherapy for patients with hepatocellular carcinoma.

B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cancer Vaccines ; immunology ; Carcinoma, Hepatocellular ; pathology ; Cell Differentiation ; Cell Line, Tumor ; Cell Proliferation ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; metabolism ; Dependovirus ; genetics ; Hep G2 Cells ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Liver Neoplasms ; pathology ; Peptide Fragments ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism ; pathology ; Transfection ; alpha-Fetoproteins ; genetics

To develop a Huntington’s disease(HD) cell model in vitro to screen drugs targeting the aggregation of polyQ,different length of CAG repeat fragments were amplified by random primer PCR, identified by DNA sequencing and were fused to the N-terminus of CAT in the pCAR system respectively which had been constructed and identified before. Recombinant plasmids were transformed into and induced to express in the host E.coli. SDS-PAGE and chloramphenicol resistance test were done to determine the solubility of the polyQ and chloramphenicol resistance levels of the fusions. With different length of CAG repeat fragments cloned and expressed in the CAT-fusion protein reporting system, it is found that when the length of the fragments increased over 40, their encoding polyQ expressed as insoluble protein and chloramphenicol resistance levels are lower, while under 40, the polyQ expressed as soluble ones and chloramphenicol resistance levels are higher. A in vitro HD model that could minimize the pathological process of the HD thus has been developed. With which by measure the recombinant bacteria’s resistance to chloramphenicol, the polyQ’ solubility and folding state in vitro by quality and quantity could be determined. Thus this model can be used to screen drugs or bioactivity materials that can inhibit aggregation of the polyQ, which thereby shedding new light on the prevent, diagnosis and therapy of HD.