Activated leukocyte cell adhesion molecule (ALCAM) is a glycoprotein of the immunoglobulin superfamily that has been implicated in the processes of cell adhesion and migration. The current study ...examines the importance of ALCAM in regulating HaCaT cell growth and migration and its potential to impact on wound healing. ALCAM levels were examined in a range of clinical wound and normal skin samples using Q‐PCR and immunohistochemistry. ALCAM expression was targeted in HaCaT keratinocyte cells using a hammerhead ribozyme transgene system. Subsequently, the impact of ALCAM suppression on HaCaT migration and growth was assessed. ALCAM protein was detected mainly in keratinocytes. ALCAM transcript levels were found to be significantly higher in the non‐healed chronic wound samples compared with healed samples (P = 0·026). In addition, targeting of ALCAM in HaCaT cells brought about a substantial increase in cellular migration and growth compared with HaCaT control cells.Our results suggest that ALCAM plays an important role in the migration of HaCaT keratinocyte cells. The data also suggests that higher levels of ALCAM may impair healing in chronic wounds. The impact of ALCAM in wound healing may thus be somewhat due to its impact on cell migration and growth.
The objective of this fixed-dose study was to determine the efficacy and safety of a new formulation of modafinil (modafinil film-coated tablets) in children and adolescents with ...attention-deficit/hyperactivity disorder (ADHD). In addition, the effect of abrupt discontinuation of modafinil was evaluated in a 2-week observation period.
Patients aged 6 to 17 years with DSM-IV-TR-defined ADHD were randomly assigned to 7 weeks of double-blind treatment with modafinil or placebo in a 2:1 ratio, followed by abrupt discontinuation of modafinil and a 2-week, double-blind observation period in which 46% of patients receiving modafinil were switched to placebo without tapering and half continued to receive modafinil. Study drug was administered once daily and titrated over the first 7 to 9 days to daily doses of 340 mg for patients < 30 kg or 425 mg for patients > or = 30 kg. Assessment instruments included the Attention-Deficit/ Hyperactivity Disorder Rating Scale-IV (ADHD-RS-IV) School and Home Versions and Clinical Global Impressions-Improvement scale (CGI-I). The study was conducted from November 2003 to June 2004.
A total of 190 patients were randomly assigned to receive modafinil (340 mg, N = 44; 425 mg, N = 82) or placebo (N = 64). 189 patients were evaluated for safety. Modafinil significantly improved symptoms of ADHD as shown by reductions in ADHD-RS-IV School Version total scores compared with placebo at all visits (p < or = .009), including the final visit of the double-blind phase (p < .0001). With modafinil, ADHD-RS-IV School Version mean total scores changed from 37.8 at baseline to 29.3 at week 1 and 20.7 at final visit; corresponding placebo values were 36.6, 32.8, and 28.4, respectively; effect size at final visit was 0.76 (95% CI = 0.63 to 0.88). Total scores on the ADHD-RS-IV Home Version were also significantly reduced at all visits (p < or = .022) and final visit (p = .001) in patients receiving modafinil compared with those receiving placebo. Significantly higher proportions of patients receiving modafinil were rated "much improved" or "very much improved" in overall clinical condition (CGI-I) at all visits compared with patients receiving placebo (p < .001). No withdrawal symptoms were observed when modafinil was abruptly discontinued at the beginning of the final 2-week observation period. Modafinil was generally well tolerated. Insomnia, headache, and decreased appetite were the most commonly reported adverse events. Sixty-three percent of patients who received modafinil completed the study; 13% discontinued because of lack of efficacy; 10%, because of adverse events; and 13%, for other reasons (e.g., consent withdrawn, lost to follow-up).
Modafinil significantly improved symptoms of ADHD both at school and at home and was well tolerated by children and adolescents. Abrupt discontinuation of modafinil was not associated with symptoms of withdrawal or with rebound of symptoms of ADHD.
Angiogenesis is an essential event during the excessive growth and metastatic spread of solid tumours. Anti-angiogenic agents have become a new choice of therapy for patients with cancer. In the ...present study, we investigated the potential effect of Yangzheng Xiaoji, a traditional Chinese medicinal formula presently used in the treatment of several solid tumours including liver cancer and gastric cancer, on angiogenesis, in vitro. The human vascular endothelial cell line HECV was used. A Matrigel-based sandwich tubule formation assay was employed to assess in vitro angiogenesis, a colorimetric method for assessing in vitro cell growth. Electric cell-substrate impedance sensing (ECIS) was used to evaluate the adhesion and migration of endothelial cells. The effects on activation of focal adhesion kinase (FAK) were evaluated using western blotting and immunofluorescence methods. Yangzhen Xiaoji extract DME25 significantly inhibited tube formation (p=0.046 vs control). This was seen together with a concentration-dependent inhibition on cell-matrix adhesion and cellular migration. It was demonstrated that the focal adhesion kinase (FAK) inhibitor PF557328 had a significant synergistic effect on DME25-induced inhibition of cell adhesion, migration and tube formation. The study showed that DME25 inhibited the phosphorylation of FAK in endothelial cells. In conclusion, Yangzhen Xiaoji has a marked effect on angiogenesis, in vitro and that this effect is at least partly mediated by the focal adhesion kinase (FAK) pathway.
Tumor endothelial markers (TEMs) are a newly discovered family of endothelial markers associated with tumor specific angiogenesis. This study sought to examine the levels of expression (qualitatively ...and quantitatively) for TEMs in human colon cancer.
Human colorectal cancer tissues (n = 48) and normal background tissues (n = 31) were obtained after surgery. RNA was extracted from frozen sections for gene amplification. The expression of TEMs (TEM-1 to TEM-8) was assessed using RT-PCR and their transcript levels were determined using real-time-quantitative PCR (Q-RT-PCR).
TEM-1 (P = 0.01), TEM-7 (P = 0.04), TEM-7R (P = 0.03), TEM-8 (P = 0.001) significantly raised in colon cancer tissues compared with the levels detected in normal background tissues. The expressions of TEM-2 and TEM-6 were found to be not significantly different between tumor tissues and normal tissues (P > 0.05). Patients who had cancer penetrating into and through the muscularis propria of the bowel wall and developed nodal involvement (Dukes C) exhibited significantly higher levels of TEM -8 compared to patients who were node negative (P < 0.05). TEM-7 and TEM-7R showed high level of transcripts in Dukes C, but they were not statistically significant.
The level of the expression of TEM-1, TEM-7, TEM-7R and TEM-8 (but not TEM-2 and TEM-6) were associated with both nodal involvement and disease progression, and may therefore, have a prognostic value in colorectal cancer.
Repulsive guidance molecules (RGMs) are novel co-receptors of bone morphogenetic proteins (BMPs) which have been implicated in bone metastasis of cancer. This study aimed to investigate roles played ...by RGMs in breast cancer.
Expression of RGMs was examined in breast cancer cell lines using RT-PCR. The expression of RGMs in human breast cancer tissues was assessed using both quantitative PCR and immunohistochemical staining.
RGMB was detectable in both cell lines and tissues samples of breast cancer. RGMA and RGMC were expressed in the breast tissues, but were undetectable in the examined breast cancer cell lines. Furthermore, reduced expression of RGMA in breast cancer was associated with poor prognosis. RGMB transcript levels appeared to be lower in breast cancer with local recurrence and distant metastasis, but were relatively higher in the patients who died from the disease.
Aberrant expression of RGMs was indicated in breast cancer. The perturbed expression was associated with disease progression and poor prognosis.
ABSTRACT
Hepatocyte growth factor (HGF) is a cytokine known to play multiple roles during the various stages of wound healing. This study addresses the ongoing key questions regarding the role of HGF ...in wound healing, namely: are HGF and its regulators expressed differently in chronic and acute wounds? Biopsies from normal skin (n=10), acute (n=10), and chronic wounds (n=17) were analyzed by immunohistochemistry, reverse transcriptase polymerase chain reaction (RT‐PCR), and quantitative real‐time RT‐PCR for the presence of HGF, its receptor cMet, its activators and its inhibitors. Immunohistochemical staining for HGF, HGF activators, and HGF inhibitors was similar, with expression being greater in chronic wound dermis compared with acute wound dermis. While expression of cMet in chronic wound dermis was less than in the acute wound dermis. PCR quantification of these proteins showed similar trends although the differences did not reach statistical significance. The results of this study provide important data confirming the role of HGF in wound healing. In addition, we have demonstrated the aberrant expression of the HGF receptor cMet and activation inhibitors (HAI‐1 and HAI‐2) in chronic wounds. This aberrant expression may have a value in predicting the process of healing and nonhealing wounds and constitute important targets of future therapies.
Deterministic electroplating repair is a novel method for rapidly repairing the attrited parts. By the qualitative contrast and quantitative comparison, influences of the current density on ...performances of the chrome-plated layer were concluded in this study. The chrome-plated layers were fabricated under different current densities when the other parameters were kept constant. Hardnesses, thicknesses and components, surface morphologies and roughnesses, and wearability of the chrome-plated layers were detected by the Vickers hardness tester, scanning electron microscope / energy dispersive X-ray detector, digital microscope in the 3D imaging mode, and the ball-milling instrument with profilograph, respectively. In order to scientifically evaluate each factor, the experimental data was normalized. A comprehensive evaluation model was founded to quantitative analyse influence of the current density based on analytic hierarchy process method and the weighted evaluation method. The calculated comprehensive evaluation indexes corresponding to current density of 40A/dm2, 45A/dm2, 50A/dm2, 55A/dm2, 60A/dm2, and 65A/dm2 were 0.2246, 0.4850, 0.4799, 0.4922, 0.8672, and 0.1381, respectively. Experimental results indicate that final optimal option was 60A/dm2, and the priority orders were 60A/dm2, 55A/dm2, 45A/dm2, 50A/dm2, 40A/dm2, and 65A/dm2.
Changes in oxygen concentration may influence various innate characteristics of stem cells. The effects of varying oxygen concentration on human periodontal ligament stem cells (HPDLSCs) has not been ...explored, particularly under hypoxia-related conditions. First, HPDLSCs were cultured from the periodontium of human teeth using the outgrowth method. STRO-1 and CD146 expression of HPDLSCs was investigated by flow cytometry. To detect the multilineage differentiation capacities of HPDLSCs, osteogenic-like and adipogenic-like states were induced in cells. Next, HPDLSCs (passage 3) were exposed to normal oxygen (21% O2) or hypoxia (2% O2) conditions for 7 days and cell proliferation was evaluated. After culture in osteogenic medium for 7 days, osteoblastic differentiation was evaluated by semi-quantitative reverse transcription-polymerase chain reaction analysis to detect 3 osteoblastic markers: core-binding factor a 1/runt-related transcription factor 2, osteocalcin, and osteopontin. In addition, each cell group was incubated with a hydroxyapatite/tricalcium phosphate carrier and transplanted subcutaneously into the back of immunocompromised mice to investigate transplantation differences in vivo. HPDLSCs were isolated, cultured, and successfully identified. After exposure of HPDLSCs to hypoxia for 7 days, the proliferation rate was increased and showed higher osteogenic differentiation potential compared to control cells. After 12 weeks of transplantation, hypoxia-treated HPDLSCs differentiated into osteoblast-like cells that formed bone-like structures. These results suggest that oxygen concentrations affect various aspects of HPDLSC physiology and that hypoxia enhances osteogenic differentiation both in vivo and in vitro. Oxygen concentration may be a critical parameter for HPDLSCs during expansion and differentiation.
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