Deformation detection of a double-clamped beam according to ultra-weak fiber Bragg grating.

Metal beam strain detection may offer valuable insight into health monitoring for large-scale steel structures. This research presents, to the best of our knowledge, the first implementation of ultra-weak fiber Bragg gratings for the deformation detection of a double-clamped beam because of their higher multiplexing and denser detection points compared to fiber Bragg gratings. The measured values are entirely consistent with those determined by strain gauges applied for reference and demonstrate that the strain of each sensing point increases linearly with the load at the middle of the beam. The errors of different loads at different load points between inversion maximum deflection and measured displacement are less than 9.59%.

MiR-320a acts as a prognostic factor and Inhibits metastasis of salivary adenoid cystic carcinoma by targeting ITGB3.

BACKGROUND: Salivary Adenoid cystic carcinoma (SACC) patients with local invasion and lung metastasis are often resistant to conventional therapy such as operation, chemotherapy and radiotherapy. To explore the underling mechanisms, we studied the roles of miRNA in regulating invasiveness of SACC cells. METHODS: MicroRNA profiling was done in SACC cells with microarray. MiRNA mimics or antisense oligonucleotide was transfected and invasiveness of SACC cells was evaluated by adhesion assay and transwell assay. The target gene of miRNA was identified by luciferase reporter assay and "rescue" experiment. Tumor metastasis was evaluated by BALB/c-nu mice xenografts. MiRNA and its target gene expression were identified by in-situ hybridization and immunohistochemistry respectively, in 302 patients from affiliated hospitals of Sun Yat-sen University and in 148 patients from affiliated hospitals of Central South University, and correlated to the clinicopathological status of the patients. RESULTS: MiR-320a was down-regulated in high lung metastatic ACCM and SACC-LM cells compared with the corresponding low metastatic ACC2 and SACC-83 cells, and inhibited adhesion, invasion and migration of SACC cells by targeting integrin beta 3 (ITGB3). In vivo, enforced miR-320a expression suppressed metastasis of SACC xenografts. In the two independent sets, miR-320a was downregulated in primary SACCs with metastasis compared to those without metastasis, and low expression of this miRNA predicts poor patient survival and rapid metastasis. Multivariate analysis showed that miR-320a expression was an independent indicator of lung metastasis. CONCLUSIONS: MiR-320a inhibits metastasis in SACCs by targeting ITGB3 and may serve as a therapeutic target and prognostic marker in salivary cancers.

The progress of PET/MRI in clinical management of patients with pancreatic malignant lesions.

Recently, the morbidity and mortality of pancreatic cancer have been increasing year by year. Because of its deep anatomical location and because most presented patients often suffer from abdominal pain or jaundice, it is difficult to diagnose pancreatic cancer at an early stage, leading to late clinical stage and poor prognosis. integrated positron emission tomography/magnetic resonance imaging (PET/MRI) fusion imaging not only has the characteristics of high resolution and multi-parameter imaging of MRI, but also combines the high sensitivity and the semi-quantitative characteristics of PET. In addition, the continuous development of novel MRI imaging and PET imaging biomarkers provide a unique and precise research direction for future pancreatic cancer research. This review summarizes the value of PET/MRI in the diagnosis, staging, efficacy monitoring, and prognosis evaluation of pancreatic cancer, and prognosis for developing emerging imaging agents and artificial intelligence radiomics in pancreatic cancer.

Fractal Permeability Model of Newtonian Fluids in Rough Fractured Dual Porous Media.

Based on the statistical self-similar fractal characteristics of the microstructure of porous media, the total flow rate and permeability of Newtonian fluids in the rough fracture network and rough matrix pores are derived, respectively. According to the connection structure between fractures and pores, the permeability analysis model of fluids in a matrix-embedded fracture network is established. The comparison between the predicted values of the model and the experimental data shows that the predicted values of the permeability of the rough fracture network and the rough matrix pores decrease with the increase in the relative roughness of the fractures and matrix pores, and are lower than the experimental data. Meanwhile, the predicted total flow rate of a rough fractured dual porous media is lower than that of a smooth fractal model and experimental data. In addition, it is also found that the larger the average inclination angle and the relative roughness of the fracture network, the smaller the permeability of the fractured dual porous media, and the relative roughness of the fracture network has a far greater influence on fluid permeability in the fractured dual porous media than the relative roughness of the matrix pores.

Biocompatibility and Angiogenic Effect of Chitosan/Graphene Oxide Hydrogel Scaffolds on EPCs.

Angiogenesis in the field of tissue engineering has attracted significant attention. Graphene oxide has become a promising nanomaterial in tissue engineering for its unique biochemical properties. Therefore, herein, a series of chitosan (CS)/graphene oxide (GO) hydrogel scaffolds were synthesized by crosslinking CS and GO at different concentrations (0.1, 0.5, and 1.0 wt.%) using genipin. Compared with the CS hydrogel scaffolds, the CS/GO hydrogel scaffolds have a better network structure and mechanical strength. Then, we used endothelial progenitor cells (EPCs) extracted from human umbilical cord blood and cocultured these EPCs with the as-prepared scaffolds. The scaffolds with 0.1 and 0.5 wt.%GO showed no considerable cytotoxicity, could promote the proliferation of EPCs and tube formation, and upregulated the expressions of CD34, VEGF, MMP9, and SDF-1 in EPCs compared to the case of the scaffold with 1.0 wt.%GO. This study shows that the addition of graphene oxide improves the structure of chitosan hydrogel and enhances the proliferation activity and angiogenic capacity of EPCs.

The role of PHF8 and TLR4 in osteogenic differentiation of periodontal ligament cells in inflammatory environment.

BACKGROUND: Histone methylation is considered to play an important role in the occurrence and development of periodontitis. Plant homeodomain finger protein 8 (PHF8), a histone demethylase, has been shown to regulate inflammation and osteogenic differentiation of bone marrow stromal cells (BMSCs). This study aimed to detect the functions of PHF8 and TLR4 in osteogenic differentiation in an inflammatory environment induced by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS) METHODS: A periodontitis mouse model was established, and the mice were treated with TAK-242. Immunohistochemical staining was used to detect the expression of PHF8 in periodontal tissue. Periodontal ligament cells (PDLCs) were treated with mineralization induction medium supplemented with Pg-LPS and/or TAK-242, and a Cell Counting Kit-8 (CCK-8) assay was used to detect the proliferation of PDLCs. Real-time PCR and western blotting were used to detect the mRNA and protein expression levels, respectively, of PHF8, toll-like receptor 4 (TLR4) and the other osteogenic markers alkaline phosphatase (ALP), osteocalcin (OCN), Special AT-rich sequence-binding protein 2 (Satb2) and Runt-related transcription factor 2 (Runx2) RESULTS: Periodontitis reduced PHF8 expression in periodontal tissue, and TAK-242 partially reversed this downregulation. An in vitro experiment revealed that the mRNA and protein expression levels of PHF8 were significantly upregulated during the osteogenic differentiation of PDLCs. Alizarin red staining showed that the mineralized nodules of PDLCs in osteogenic induction group were more than those in control group. Real-time PCR and western blot results indicated that Pg-LPS inhibited PHF8 expression and upregulated TLR4 expression in PDLCs. TAK-242 inhibited TLR4 and partially reversed the inhibition of PHF8 expression and osteogenic differentiation induced by Pg-LPS in PDLCs CONCLUSION: PHF8 and TLR4 play important roles in periodontitis. Pg-LPS inhibits the expression of PHF8 via upregulation of TLR4 and might further inhibit the osteogenic differentiation of PDLCs. However, the specific mechanisms involved remain to be explored.

Polypeptides IGF-1C and P24 synergistically promote osteogenic differentiation of bone marrow mesenchymal stem cells in vitro through the p38 and JNK signaling pathways.

OBJECTIVES: Insulin-like growth factor-1 (IGF-1) and bone morphogenetic protein 2 (BMP-2) both promote osteogenesis of bone marrow mesenchymal stem cells (BMSCs). IGF-1C, the C domain peptide of IGF-1, and P24, a BMP-2-derived peptide, both have similar biological activities as their parent growth factors. This study aimed to investigate the effects and mechanisms of polypeptides IGF-1C and P24 on the osteogenic differentiation of BMSCs. METHODS: The optimum concentrations of IGF-IC and P24 were explored. The effects of the two polypeptides on BMSC proliferation and osteogenic differentiation were examined using a CCK-8 assay, flow cytometry, alkaline phosphatase (ALP) staining, ALP activity assay, alizarin red S staining, qPCR, and Western blotting. In addition, specific pathway inhibitors were utilized to explore whether the p38 and JNK pathways were involved in this process. RESULTS: The optimal concentration of both polypeptides was 50 µg/ml. IGF-1C and P24 synergistically promoted BMSC proliferation, increased ALP activity and calcified nodule formation, upregulated the mRNA and protein levels of Osx, Runx2, Ocn, Opn, and Col1a1, and improved the phosphorylation levels of p38 and JNK proteins. Inhibition of the pathways significantly reduced p38 and JNK activation and blocked Runx2 expression while inhibiting ALP activity and calcified nodule formation. CONCLUSIONS: These findings suggest that IGF-1C and P24 synergistically promote the osteogenesis of BMSCs through activation of the p38 and JNK signaling pathways.

Retraction of "Biological Effects of Titanium Surface Charge with a Focus on Protein Adsorption".

[This retracts the article DOI: 10.1021/acsomega.0c02518.].

Range of turbulence-independent propagation and Rayleigh range of partially coherent beams in atmospheric turbulence.

Two characteristic distances for partially coherent beams propagating in atmospheric turbulence have been proposed. The turbulent Rayleigh range is used for characterizing the range over which the beams propagate in turbulence without spreading appreciably; i.e., the concept of the well-known Rayleigh range in free space is extended to the case of turbulence. In this paper the range of turbulence-independent propagation of the beams, in contrast to similar characteristic distances in previous published works, is based on the formula of the beam propagation factor (M(2) factor) and is used for describing the range over which the spatial and angular spreading and the M(2) factor increase due to turbulence are sufficiently small and negligible. Several simple formulas used for calculating the approximate values of these distances are given, and the formulas are applied to Gaussian Schell-model (GSM) beams and illustrated by examples. Furthermore, as a typical example, the effect of the angular spread of GSM beams in turbulence on a thin-lens optical system is also discussed. We show that the turbulent Rayleigh range depends on the Rayleigh range in free space, the waist width, and the spatial power spectrum of the refractive-index fluctuations of the turbulent atmosphere, and that the range of turbulence-independent propagation depends on the waist width, the initial angular spread in the waist plane, and the spatial power spectrum.

Biological Effects of Titanium Surface Charge with a Focus on Protein Adsorption.

The effect of changes in surface charge on the biological properties of implants is not clear. The objective of this study was to evaluate the biological properties of the surface of titanium sheets with different charges due to different treatment methods. Titanium sheets were sandblasted with large grit and underwent acid etching before being subsequently divided into the following groups: SLA, no further treatment; SLA-Ca2+, immersed in 1% CaCl2 solution; SLA-NaCl, immersed in saline; and SLA-Ca2+-NaCl, immersed in 1% CaCl2 solution followed by saline. Surface characteristics were evaluated using field-emission scanning electron microscopy with energy-dispersive spectrometry, surface profilometry, and contact angle assays. Additionally, we used a ζ-potential analyzer to directly measure the electrostatic charge on the different group surfaces. The effect of changes in the Ti surface on biological processes after different treatments was determined by analyzing fibronectin adsorption, osteoblast-like MG63 cell adhesion and proliferation, and the expression of osteogenesis-related genes. Compared to the SLA surface, the other three groups contained corresponding trace elements because they were soaked in different liquids; the contact angles of the three groups were not significantly different, but they were significantly smaller than that of the SLA group; and there was no change in the surface topography or roughness. Furthermore, the SLA-Ca2+ group had a significantly reduced negative charge compared to that of the other three groups. There were no differences between the SLA-NaCl and SLA-Ca2+-NaCl groups in terms of negative charge, and the SLA group surface carried the most negative charge. Fibronectin adsorption capacity and cytological performance testing further showed that the SLA-Ca2+ group had the most significant change, followed by the SLA-NaCl and SLA-Ca2+-NaCl groups; the SLA group had significantly lower capacity and performance than the other three groups. These results suggest that the surface charge of the titanium sheet changed when immersed in different liquids and that this treatment enhanced biocompatibility by reducing the electrostatic repulsion between biomaterials and biomolecules.

[Biocompatibility of poly-L-lactic acid/Bioglass-guided bone regeneration membranes processed with oxygen plasma].

OBJECTIVE: To prepare and characterize a nano-scale fibrous hydrophilic poly-L-lactic acid/ Bioglass (PLLA/BG) composite membrane and evaluate its biocompatibility as a composite membrane for guiding bone regeneration (GBR). METHODS: PLLA/BG-guided bone regeneration membrane was treated by oxygen plasma to improved its hydrophilicity. The growth of MG-63 osteoblasts on the membrane was observed using Hoechst fluorescence staining, and the biocompatibility of the membrane was evaluated by calculating the cells adhesion rate and proliferation rate. Osteogenesis of MG-63 cells was assessed by detecting alkaline phosphatase (ALP), and the formation of calcified nodules and cell morphology changes were observed using scanning electron microscope (SEM). RESULTS: The cell adhesion rates of PLLA/BG-guided bone regeneration membrane treated with oxygen plasma were (30.570±0.96)%, (47.27±0.78)%, and (66.78±0.69)% at 1, 3, and 6 h, respectively, significantly higher than those on PLLA membrane and untreated PLLA/BG membrane (P<0.01). The cell proliferation rates on the 3 membranes increased with time, but highest on oxygen plasma-treated PLLA/BG membrane (P<0.01). Hoechst fluorescence staining revealed that oxygen plasma treatment of the PLLA/BG membrane promoted cell adhesion. The membranes with Bioglass promoted the matrix secretion of the osteoblasts. Under SEM, the formation of calcified nodules and spindle-shaped cell morphology were observed on oxygen plasma-treated PLLA/BG membrane. CONCLUSION: Oxygen plasma-treated PLLA/BG composite membrane has good biocompatibility and can promote adhesion, proliferation and osteogenesis of the osteoblasts.

Preparation and characterization of genipin-cross-linked silk fibroin/chitosan sustained-release microspheres.

We report the effects of distinct concentrations of genipin and silk fibroin (SF):chitosan (CS) ratios on the formation of SF-CS composite microspheres. We selected microspheres featuring an SF:CS ratio of 1:1, encapsulated various concentrations of bovine serum albumin (BSA), and then compared their encapsulation efficiency and sustained-release rate with those of pure CS microspheres. We determined that the following five groups of microspheres were highly spherical and featured particle sizes ranging from 70 µm to 147 µm: mass ratio of CS:SF =1:0.5, 0.1 g or 0.5 g genipin; CS:SF =1:1, 0.05 g or 1 g genipin; and CS:SF =1:2, 0.5 g genipin. The microspheres prepared using 1:1 CS:SF ratio and 0.05 g genipin in the presence of 10 mg, 20 mg, and 50 mg of BSA exhibited encapsulation efficiencies of 50.16%±4.32%, 56.58%±3.58%, and 42.19%±7.47%, respectively. Fourier-transform infrared spectroscopy (FTIR) results showed that SF and CS were cross-linked and that the α-helices and random coils of SF were converted into ß-sheets. BSA did not chemically react with CS or SF. Moreover, thermal gravimetric analysis (TGA) results showed that the melting point of BSA did not change, which confirmed the FTIR results, and X-ray diffraction results showed that BSA was entrapped in microspheres in a noncrystalline form, which further verified the TGA and FTIR data. The sustained-release microspheres prepared in the presence of 10 mg, 20 mg, and 50 mg of BSA burst release 30.79%±3.43%, 34.41%±4.46%, and 41.75%±0.96% of the entrapped BSA on the 1st day and cumulatively released 75.20%±2.52%, 79.16%±4.31%, and 89.04%±4.68% in 21 days, respectively. The pure CS microspheres prepared in the presence of 10 mg of BSA burst release 39.53%±1.76% of BSA on the 1st day and cumulatively released 83.57%±2.33% of the total encapsulated BSA in 21 days. The SF-CS composite microspheres exhibited higher sustained release than did the pure CS microspheres, and thus these composite microspheres might function as a superior drug carrier.

Characterization of Silk Fibroin/Chitosan 3D Porous Scaffold and In Vitro Cytology.

Bone tissue engineering is a powerful tool to treat bone defects caused by trauma, infection, tumors and other factors. Both silk fibroin (SF) and chitosan (CS) are non-toxic and have good biocompatibility, but are poor biological scaffolds when used alone. In this study, the microscopic structure and related properties of SF/CS composite scaffolds with different component ratios were examined. The scaffold material most suitable for osteoblast growth was determined, and these results offer an experimental basis for the future reconstruction of bone defects. First, via freeze-drying and chemical crosslinking methods, SF/CS composites with different component ratios were prepared and their structure was characterized. Changes in the internal structure of the SF and CS mixture were observed, confirming that the mutual modification between the two components was complete and stable. The internal structure of the composite material was porous and three-dimensional with a porosity above 90%. We next studied the pore size, swelling ratio, water absorption ratio, degradation and in vitro cell proliferation. For the 40% SF-60% CS group, the pore size of the scaffold was suitable for the growth of osteoblasts, and the rate of degradation was steady. This favors the early adhesion, growth and proliferation of MG-63 cells. In addition to good biocompatibility and satisfactory cell affinity, this material promotes the secretion of extracellular matrix materials by osteoblasts. Thus, 40% SF-60% CS is a good material for bone tissue engineering.

Silencing Dicer expression enhances cellular proliferative and invasive capacities in human tongue squamous cell carcinoma.

microRNAs (miRNAs) are aberrantly expressed in cancer. An enzyme essential for miRNA processing is Dicer, whose expression is deregulated in diverse types of cancer and correlates with tumor progression. However, whether the regulation of Dicer expression affects tongue squamous cell carcinoma is unknown. In the present study, we investigated how silencing the expression of Dicer alters cell proliferation, cell cycle patterns, and cell migration and invasion in the Tca-8113 tongue squamous cell carcinoma cell line. Dicer expression levels were determined using quantitative PCR and western blot analysis in normal oral gingival epithelial cells and in two tongue squamous cell carcinoma lines, Tca-8113 and UM-1. Tca-8113 cells were transfected with Dicer siRNA or a negative control siRNA. Cell proliferation was determined using the MTT assay and the cell cycle was examined using flow cytometry. Cell migration and invasion changes were evaluated using wound-healing, adherence and Transwell assays. Dicer was expressed at lower levels in the tongue squamous cell carcinoma cell lines Tca-8113 and UM-1 compared to normal gingival epithelial cells, and less Dicer was expressed in UM-1 cells compared to Tca-8113 cells. Notably, Tca-8113 cells transfected with Dicer siRNA had significantly higher proliferative and invasive abilities than cells transfected with the negative control siRNA or non-transfected cells. Silencing Dicer may promote the progression of tongue squamous cell carcinoma. Dicer could serve a promising biomarker and a potential therapeutic target for tongue squamous cell carcinoma.

[Preparation and characterization of genipin-crosslinked silk fibroin/chitosan controlled-release microspheres].

OBJECTIVE: To investigate the property of genipin-crosslinked silk fibroin(SF)/chitosan(CS) microspheres for slow releasing of bovine serum albumin (BSA). METHODS: BSA-loaded genipin-crosslinked SF/CS microspheres were prepared by emulsion cross-linking technique. The micropheres were observed for surface morphology and size distribution under scanning electron microscope (SEM), and X-ray diffractometry (XRD) and fourier transform infrared spectroscopy (FTIR) were used to analyze their structural characteristics. BCA method was used for determining the drug entrapment, loading rate and cumulative drug release in 21 days. RESULT: The microspheres were spherical and showed a smooth surface with an average diameter of 7.84∓0.97 µm. The drug entrapment efficiency of the microspheres was (50.16∓4.32)% with a drug loading ratio of (1.25∓0.11)% and a cumulative release of the total drug of (75.2∓2.53)% in 21 days. CONCLUSION: Genipin-crosslinked SF/CS microspheres have a high drug entrapment efficiency and possess good capacity of sustained drug release.

[Intratumor injection of recombinant attenuated salmonella carrying Mycobacterium tuberculosis heat shock protein 70 and herpes simplex virus thymidine kinase genes to suppress murine melanoma growth].

OBJECTIVE: To study the effection of suppression murine melanoma growth by Intratumor injection of recombinant attenuated salmonella carrying heat shock protein 70 and herpes simplex virus thymidine kinase genes. METHODS: Plasmids PCMV-mtHSP70-IRES-TK were electro-transferred into salmonella typhimurium SL7207 to construct recombinant salmonella typhimurium. In vivo, Recombinant bacteria were injected into the mouse melanoma and the antitumor effection was observed. The survival period was recorded and safety analysis for this vaccine in each group. RESULTS: In vivo, the mtHSP70/HSV-tk recombinant bacteria can suppress tumor growth significantly and extend survival. After recombinant Salmonella, 10(9) CFU/mL, was administered as an intratumoral injection, No diarrhea were observed. During therapy, body weight did not change markedly. CONCLUSION: Results of the animal experiment suggests intratumor injection of recombinant attenuated salmonella typhimurium containing mtHSP70 and HSV-tk genes, has targeting ability against B16 tumor cell and could significantly inhibit tumor growth .

[Tyrosine kinase 2 with immunoglobulin-like and epidermal growth factor homology domains RNA interference inhibit the proliferation of human umbilical vein endothelial cells].

OBJECTIVE: The purpose of this study was to investigate the regulatory role of tyrosine kinase 2 with immunoglobulin-like and epidermal growth factor homology domains (Tie2) on apoptosis and proliferation in the endothelial cells. METHODS: RNA interference (RNAi) technique was used to silence Tie2 gene expression by transfecting an expression vector containing short hairpin RNA(shRNA) for Tie2 into human umbilical vein endothelial cells (HUVECs). Real time quantitation reverse transcriptase polymerase chain reaction (QRT-PCR) and Western blot were used to monitor Tie2 mRNA, as well as protein expression. The proliferation of HUVECs was examined by methyl thiazolyl tetrazolium (MTT), and the apoptosis was detected under microscope. HUVECs transfected with pGenesil-hk was negative control, and HUVECs transfected with nothing was empty control. RESULTS: Tie2 mRNA expression was down-regulated 24 h and 48 h after transfection, and Tie2 protein expression was significantly down-regulated at 24 h and 48 h (P< 0.05), especially 48 h after transfection. The apoptosis rate was conspicuously higher in experimental group than in negative control and empty control group after 48 h (P<0.05). The growth monitoring showed that proliferation was also markedly inhibited in experimental group (P<0.05) compared with two control groups. CONCLUSION: Down-regulated expression of Tie2 by RNAi can promotes apoptosis of HUVECs and has an anti-proliferation activity effect on them.

Suppression of murine melanoma growth by a vaccine of attenuated Salmonella carrying heat shock protein 70 and Herpes simplex virus-thymidine kinase genes.

Attenuated Salmonella can invade tumor cells and acts as a eukaryotic expression vector for gene propagation. We constructed a bi-gene, eukaryotic co-expression DNA vaccine of Mycobacterium tuberculosis heat shock protein 70 (mtHSP70) and Herpes simplex virus-thymidine kinase (HSV-tk) and used attenuated Salmonella as a vector to treat murine melanoma. In vitro, recombinant Salmonella can carry plasmid stably and can invade into the cytoplasm of B16 tumor cells expressing the protein of the mtHSP70/HSV-tk gene by Western blot assay. In vivo, after the recombinant Salmonella was injected into tumors, the HSV-tk precursor drug ganciclovir (GCV) was administered to start the HSV-tk killing of tumor cells. We found that the mtHSP70/HSV-tk recombinant bacteria can raise CD8+ T lymphocytes in peripheral blood by flow cytometry and in tumor tissues by immunofluorescence detection, increase IFN­Î³ contents in tumor tissue by ELISA and significantly suppress tumor growth.

[Culture and identify the human umbilical vein endothelial cells and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains in the cells].

OBJECTIVE: To study the cultural method and identification of human umbilical vein endothelial cells (HUVECs), and investigate the expression of tyrosine kinase-2 with immunoglobulin-like and epidermal growth factor homology domains(Tie-2) in HUVECs. METHODS: HUVECs were isolated from umbilical veins by the technique of irrigative digestion, and were cultivated in plates. The cells were identified by VIII monoclonal antibody. Tie-2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and SABC immunocytochemistry. RESULTS: HUVECs could adhere to the plates completely after 24 hours, and confluence a monolayer 4-5 days later. The band of Tie-2 mRNA was obviously and the expression of Tie-2 protein was strongly positive by immunocytochemistry in HUVECs. The positive rate was over 85%. CONCLUSION: Highly purified endothelial cells were isolated. And there were overexpression of Tie-2 in HUVECs.

Inhibition of VEGF expression in tongue squamous cancer cells via RNA interference silencing of iNOS gene.

The purpose of this study was to investigate the regulatory role of the inducible nitric oxide synthase (iNOS) gene on vascular endothelial growth factor (VEGF) expression in oral squamous cancer cells. The RNA interference (RNAi) technique was used to silence iNOS gene expression by transfecting an expression vector containing short hairpin RNA (shRNA) for iNOS into Tca8113 tongue squamous cancer cells using cationic liposomes. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blotting were used to monitor iNOS and VEGF mRNA, as well as protein expression. iNOS mRNA expression was significantly downregulated 24 and 36 h after transfection, and iNOS protein expression was significantly downregulated at 36 and 48 h (P<0.05 versus control), showing that effective silencing was achieved. VEGF mRNA was significantly decreased 24 and 36 h post-transfection, and VEGF protein expression was significantly decreased at 36 and 48 h (P<0.05). RNAi can decrease iNOS gene expression and achieve a gene silencing effect. iNOS gene silencing reduces VEGF expression levels in Tca8113 cells. Thus, there is a relationship between iNOS and VEGF expression in tongue squamous cancer cells.