Stable expression of antisense urokinase mRNA inhibits the proliferation and invasion of human hepatocellular carcinoma cells

Urokinase-type plasminogen activator (u-PA) plays a key role in malignant tumor behavior. We have previously shown that the expression of high levels of u-PA mRNA in human hepatocellular carcinoma (HCC) biopsies was inversely correlated with the survival of the patients. In order to evaluate the involvement of u-PA in the invasive and infiltrating properties of HCC cells, the SKHep1C3 cell line was stably transfected with an expression vector containing the 5' portion (257 bp) of u-PA cDNA in the antisense orientation. u-PA mRNA expression and its protein level and enzymatic activity were specifically inhibited in the antisense transfectants. A comparable inhibition of the u-PA receptor (u-PAR) mRNA and protein was also evidenced in the antisense transfected cells compared with the control ones. At the functional level, the SKHep1C3-AS cells showed a significant reduction in proliferation, Matrigel invasion, and motility assays compared to parental and vector-alone cells. These results indicate that u-PA is an essential factor in the growth and invasiveness of human hepatocarcinoma cells. Antisense u-PA strategy might be a potential approach to reduce tumor growth as well as the invasive capacity of the malignant cells in HCC.     

Temporal facelift and corrugator resection

On a path analogous to other surgical disciplines, the progression of facial cosmetic surgery has trended toward shorter scars and less invasive techniques. This article outlines an approach to facelifting that limits the scar to the temporal scalp, preauricular area, and retroauricular sulcus. It will enable the reader to identify those patients who are candidates for a less invasive procedure compared with the conventional facelift approach. Specifically, the temporal facelift addresses the lateral brow, the middle one third of the face, and the upper neck. In addition, this article examines the various and current methods for treatment of the glabellar area of the forehead. Medical and surgical options are outlined for the correction of those rhytids induced by corrugator and procerus muscle activity.     

Smokeless tobacco and nicotine bring about excessive cytokine responses of murine memory T-cells

To determine the influence of smokeless tobacco (ST) and nicotine on the cytokine phenotype of memory T-cells, splenic mononuclear cells (SPM) were exposed to 1:10(2) or 1:10(3) dilutions of ST extract (ST-SPM), 10 or 100 microg/ml nicotine (NIC-SPM), or medium (CON-SPM) during 4 days of stimulation with anti-CD3. SPM were then washed extensively to remove residual ST or nicotine and restimulated with anti-CD3 and anti-CD28 in the absence of ST or nicotine. Expression of IL-2, IL-4, IL-10 and IFN-gamma protein and mRNA levels after 4 days of primary stimulation and after 24 and 48 h of restimulation was evaluated using ELISA and RT-PCR, respectively. After 4 days of primary stimulation, SPM exposed to 100 microg/ml nicotine sustained expression of IL-2, IFN-gamma, IL-10 and IL-4 mRNA as opposed to CON-SPM. Restimulation of CON-SPM resulted in maximum re-expression of cytokine mRNA at 24 h and a decline by 48 h. Restimulated NIC-SPM in the absence of nicotine delayed maximal re-expression of IL-2, IFN-gamma, IL-10 and IL-4 mRNA until 48 h. Heightened expression of cytokine mRNA at 48 h was paralleled by a small but significant increase in production of IFN-gamma, IL-4 and IL-10 protein by NIC-SPM as measured by ELISA. In contrast, ST-SPM did not exhibit residual expression of cytokine mRNA after 4 days of primary stimulation. Like NIC-SPM, however, restimulated ST-SPM exhibited maximum IL-2, IL-4, IFN-gamma, and IL-10 mRNA at 48 h. Heightened re-expression of cytokine mRNA at 48 h by ST-SPM was paralleled by increased production of IL-2, IFN-gamma, IL-4 and IL-10 protein. These results indicate that exposure of T-cells to nicotine, but not ST, during a primary immune response result in inordinate cytokine expression after 4 days. In addition, memory T-cells initially exposed to nicotine or ST during a primary immune response, exhibit excessive cytokine expression when T-cells are restimulated in the absence of nicotine or ST. pharmacology.     

Spontaneous dissection of the cervical internal carotid artery: correlation of arteriography, CT, and pathology

Spontaneous dissection of the internal carotid artery is being recognized as a more frequent cause of acute neurologic deficit, particularly in young persons. Saccular pseudoaneurysm formation may be an associated finding, especially in the presence of tortuosity (coiling) of the cervical internal carotid artery. Of eight patients with nine vessels demonstrating internal carotid artery dissection on arteriography, pseudoaneurysms were found in five arteries. Four of the five pseudoaneurysms occurred in tortuous (coiled) arterial segments. Thin-section contrast-enhanced dynamic incremental CT showed close agreement with the findings on selective arteriography and provided additional information on the presence and configuration of arterial wall thickening as well as the extent of the pseudoaneurysm. Our experience indicates that CT may play an important role in the diagnosis, management, and follow-up of this lesion.     

Headache in the athlete and radiographic evaluation

A deliberate attempt is made to stress the nonspecificity of headache and the complexity of its work-up. The choice of radiographic studies for its evaluation should be governed by the strongly considered etiology. This article deals with the most likely causes of headaches in an athlete. The merits and limitations of currently available radiographic modalities to detect headache-producing diseases are broadly discussed.     

Small interfering RNA urokinase silencing inhibits invasion and migration of human hepatocellular carcinoma cells

The serine protease urokinase-type plasminogen activator (u-PA) is involved in a variety of physiologic and pathological processes; in particular, u-PA mRNA is up-regulated in human hepatocellular carcinoma (HCC) biopsies and its level of expression is inversely correlated with patients' survival. To determine the role of u-PA in the invasiveness properties of HCC, we successfully down-regulated u-PA by RNA interference (RNAi) technology, in an HCC-derived cell line at high level of u-PA expression. RNAi is a multistep process involving generation of small interfering RNAs (siRNA) that cause specific inhibition of the target gene. SKHep1C3 cells were transfected with a U6 promoter plasmid coding for an RNA composed of two identical 19-nucleotide sequence motifs in an inverted orientation, separated by a 9-bp spacer to form a hairpin dsRNA capable of mediating target u-PA inhibition. Stable transfectant cells showed a consistently decreased level of u-PA protein. In biological assays, siRNA u-PA-transfected cells showed a reduction of migration, invasion, and proliferation. In conclusion, u-PA down-regulation by RNAi technology decreases the invasive capability of HCC cells, demonstrating that stable expression of siRNA u-PA could potentially be an experimental approach for HCC gene therapy.