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Background: Tongue cancer metastasis is mainly through blood stream and possibly associated with tumor cell-induced platelet aggregation (TCIPA)., Methods: Platelet aggregation was induced by different amounts of SAS tongue cancer cells with/without inhibitors and the latent period for induction of platelet aggregation was recorded. Gene expression was analyzed by reverse transcriptase-polymerase chain reaction., Results: SAS cells (4 x 10(4) to 1 x 10(6) cells/ml) induced platelet aggregation in a cell density-dependent manner. The latent period for induction of platelet aggregation reduced from 11.3 min (2 x 10(5) cells/ml) to 0.9 min (5 x 10(5) cells/ml). The extent of platelet aggregation increased from 39% to 76% by 2 x 10(5) and 5 x 10(5) SAS cells. Pre-treatment of SAS cells with aspirin showed little effect on its induction of platelet aggregation. SAS cells expressed tissue factor (TF) mRNA and the SAS cells-induced TCIPA was inhibited by TF neutralization antibody (5-20 microg/ml), heparin (5-10 U/ml), Hirudin fragment 54-65 (50 microg/ml) and D-Phenylalanyl-L-prolyl-L-arginine chloromethyl ketone. But areca nut (AN, a betel quid component known to generate reactive oxygen species (ROS)) extract showed little effect on TF expression in SAS cells. Pre-treatment with U73122 and 2-aminoethoxydiphenylborate inhibited SAS-induced TCIPA. Interestingly, catalase suppressed SAS cells-induced TCIPA, whereas AN extract enhanced this event., Conclusions: These results suggest that tongue cancer cells may induce TCIPA and enhance tumor metastasis. SAS-induced TCIPA is related to TF secretion, thrombin generation and associated with Phospholipase C-Inositol triphosphate signaling and ROS production. Betel quid chewing may potentially promote tongue cancer metastasis.

This article has a special focus on the broad range of innovative nanoprobes for signal amplification and new generations of bioassays. Advances in functionalizing gold nanoparticles with oligonucleotides speed up the development of a series of new nucleic acid assays. A biobarcode assay allows signal amplification by utilizing antibody-coated magnetic beads to concentrate the analytes and antibody-coated gold nanoparticle probes to carry a large number of oligonucleotides. Novel signal-amplification technologies, based on either new classes of nanoprobes consisting of releasable fluorophores or with aggregation-induced emission features, can also improve the sensitivity of bioassays. Advances in synthesis and biofunctionalization of quantum dots with unique properties have generated increasingly widespread applications in DNA sorting, multiplexing bioassays and fluorescence resonance energy transfer-based sensing. Ingenious nanoprobes in bioassays can offer PCR-like sensitivity, high selectivity, capacity for massive multiplexing, time efficiency and, most importantly, the ability to be performed at the point- of-care.

A unique approach of developing a bar code version of lateral-flow enzymatic-based assay for the semi-quantification of hydrogen peroxide is described. The proposed assay system is mainly composed of a goat anti-mouse IgG-horseradish peroxidase conjugate (Gt anti-M IgG-HRP)-coated nitrocellulose (NC) membrane and a peroxidase substrate pad. Unlike the bar code immunochromatographic assay which depends on the stepwise capture of analyte, the principle of enzyme-based bar code lateral-flow assay is based on the different reaction time on successive lines due to the delay in 3,3',5,5'-tetramethylbenzidine (TMB) release. Hydrogen peroxide (H(2)O(2)) acts as a limiting factor which controls the rate of the enzymatic conversion of TMB to blue color complex. The system expresses the concentration of H(2)O(2) in micromole range as three distinct ladder bars in 9 min therefore without the need of any reading device. The major advantages of this assay are its easily readable result, and also its simplicity and low-cost in production offers a cheaper alternative for testing those expensive biosensors might not be available to the third world countries. By incorporating with H(2)O(2)-generating oxidoreductases, the assay can be further extended to detect a variety of analytes with clinical and environmental importance. Glucose was chosen to be the model analyte where the proposed system gave signal response at between 5 microM and 100 microM.

The cytoplasmic enzyme dihydrodiol dehydrogenase (DDH) plays an important role in detoxification. Patients with DDH overexpression have a significantly higher incidence of early tumor recurrence and distant metastasis. This study evaluated the correlation between clinicopathological data and DDH expression and the prognostic significance of DDH expression in patients with resected gastric cancer. Between January 1998 and September 2004, we retrospectively enrolled 81 patients who received surgical treatment for gastric cancer. Pathology samples were immunostained with monoclonal antibody to DDH. The relationship between DDH expression and clinicopathological data (age, gender, histological type, stage) was analyzed by chi-square analysis. Survival curves were plotted using the Kaplan-Meier method and compared using a log-rank test. The overexpression rate of DDH was 41.9%. Of patients with overexpressed DDH, 13% had stage I, 24% had stage II, 52% had stage III, and 78% had stage IV tumors. Among patients who died, DDH expression level differed significantly between high and low-expression groups (P = 0.042). Survival was significantly better in patients with low DDH expression (P = 0.048). Thus, DDH expression may be useful in identifying high-risk gastric cancer patients and distinguishing future candidates for curative and palliative treatment.

Objectives: In this study we aimed to investigate the roles of neopterin, C-reactive protein (CRP) and the CRP to neopterin (C/N) ratio to differentiate bacterial from viral aetiology in patients with suspected acute respiratory tract infections (ARTIs) presenting to the emergency department (ED)., Methods: Serum was taken from five hundred and sixty-one patients and used to measure neopterin and CRP levels. The primary outcome was bacterial or viral infection based on positive bacterial culture and positive viral serology. Patients were classified as either: group 1 with positive bacterial culture and mixed bacterial/viral growth; group 2 with virological aetiology, and group 3 with unknown microbiological aetiology., Results: The median of the C/N ratio was 10 times higher in patients with bacterial aetiology than with viral aetiology (12.5 vs 1.2mg/nmol; P<0.0001), and 42 times higher than those in healthy subjects (12.5 vs 0.3mg/nmol; P<0.0001). The area under the receiver-operator characteristic curve for the C/N ratio was 0.840 (0.783-0.898; P<0.05). A cut-off value of "C/N ratio >3" for ruling in/out bacterial/viral infection yielded optimal sensitivity and specificity of 79.5% and 81.5% respectively. A sensitivity analysis performed on all patients (including unknown aetiology) with a cut-off value of "C/N ratio >3" yields a best-case scenario for ruling in/out bacterial/viral infection with sensitivity of 93.1% and specificity of 93.0%., Conclusion: This study shows that CRP and neopterin have a role in differentiating bacterial from viral causes of ARTI, and the C/N ratio yields optimal differentiation in the ED setting.

A lateral-flow, enzyme-based, bar-code assay for creatinine employing the concept of combination of diffusion and kinetics controlled has been developed. Unlike the traditional bar-code version of immunochromatographic assay, which depends on the stepwise capture of colorimetric tracer-labeled antibody-antigen complex by the immobilized antibody on each successive line, the principle of our proposed assay is based on the delay in TMB release and its diffusion in combination with horseradish peroxidase kinetics. Hydrogen peroxide (H(2)O(2)) produced from enzymatic reactions acts as a limiting factor, which controls the rate of conversion of TMB to blue color complex. The assay takes advantage of giving ladder bar result therefore without the need of any reading device. Depending on the amount of enzymes used, the assay can be one (9 min) or two steps (19 min). The strip assay semiquantitatively measures creatinine concentrations ranging from 0 to 400 microM. Thirty urine samples and thirty serum samples were tested, and the assay showed 90.0% and 86.7% agreement compared with conventional Jaffé method, respectively. This assay provides a tool for quick identification of creatinine for patients without the requirement of any instrument.

This study focuses a novel method to remove the human pathogens cryptosporidium parvum from water by silica particles coated with functionalized self-assembled monolayers. The results of this investigation clearly show that the pathogen can efficiently and completely be removed at pH ranges of drinking water by stirring the coated particles in the contaminated water for up to 60 min and finally filtrating the powder. The removal is believed to be caused by electrostatic attraction and immobilization of pathogen on the surface of the particles. At higher pH vales, even chemisorption may occur.

Left ventricular (LV) systolic dyssynchrony is an important pathologic mechanism in patients with heart failure (HF). However, the prevalence of intraventricular dyssynchrony in patients with different LV ejection fractions (EFs) is unknown. This study evaluated 402 consecutive patients with HF (mean age 64.99 +/- 13.15 years, 72.4% men) and 120 healthy controls. Dyssynchrony indexes included the SD of the time to peak systolic velocity (Ts) in ejection phase in the 12-segmental model (Ts-SD) and the difference in Ts between basal septal and basal lateral segments (Ts-Septal-Lateral) using tissue Doppler imaging. Patients were divided into 3 groups according to LVEF (LVEF <20%, >20% to 35%, and >35% to 50%) and compared with healthy controls. Both indexes were significantly higher in all 3 LVEF groups compared with controls (p <0.0001). Based on the established cut-off values, systolic dyssynchrony was equally prevalent in all 3 LVEF groups and was 67%, 62%, and 55% using Ts-SD and 38%, 36%, and 35% using Ts-Septal-Lateral, respectively. However, the prevalence of systolic dyssynchrony was higher using Ts-SD than Ts-Septal-Lateral (chi-square = 94.43, p <0.001). Conversely, the prevalence of electrical dyssynchrony, defined as a >120-ms QRS duration, decreased significantly with increasing LVEF (44%, 35%, and 16%; chi-square 5.60, p <0.001). In conclusion, the prevalence of mechanical systolic dyssynchrony was independent of severity of LV systolic dysfunction. This may implicate the potential role of cardiac resynchronization therapy for those with LVEF of 35% to 50%, in particular when systolic dyssynchrony is present.

In the present study, we describe an InfectCheck barcode-style lateral flow assay for semi-quantitative detection of CRP in distinguishing between bacterial and viral infections. The severity of bacterial infection can be assessed by simply counting the number of red lines developed at the CRP test zone of the test device. If only one visible line appeared at the CRP test zone, it represents a low or mild inflammation with CRP levels <10 mg/L. Two and three visible lines mean moderate (>or=10-25 mg/L) and severe (>or=25-50 mg/L) inflammations respectively while four visible lines stand for a very severe inflammation (>or=50-100 mg/L). If the visible lines become faint and the intensity of the first line is weaker than that of the control line and may even disappear, this outcome corresponds to the stage of having super severe inflammation (>or=100 mg/L). A total of 500 patients admitted to hospital through the Accident and Emergency Unit at the Prince of Wales Hospital were examined. The InfectCheck CRP barcode-style rapid test gave a high sensitivity of 88.7% and a high negative predictive value of 93.8%. This result indicates that the rapid test is reliable to exclude non-infected patients. The calculated intra- and inter-assay coefficient of variation for the five CRP concentration ranges was both within 20.0%. It is a one-step whole blood rapid test without any sample pre-treatment and the result is available within 20 min. This user friendly diagnostic tool can allow self-testing by interested individuals without any expensive reading device.

Transforming growth factor beta (TGF-beta) may regulate the biological activities of dental pulp cells. We found that human dental pulp cells expressed TGF-beta1, TGF-beta2, and a little amount of TGF-beta3 messenger RNA (mRNA). The exposure of pulp cells to TGF-beta2 induced the phosphorylation of Smad2/3, Smad1/5/8, and extracellular regulated-kinase 1/2 (ERK1/2) as observed by Western blotting. Exposure to TGF-beta2 decreased the alkaline phosphatase (ALP) mRNA expression and enzyme activity. Pretreatment of pulp cells with SB431542 (an inhibitor of TGF-beta ALK-4, ALK-5, and ALK-7 receptors) but not U0126 (a MEK1 inhibitor) prevented the inhibition of viable cell number, ALP activity, and mRNA expression by TGF-beta2 in dental pulp cells. These results suggest that TGF-beta may affect the growth and differentiation of dental pulp cells via an autocrine fashion by activation of the ALK/Smad2/3-signal transduction pathways. TGF-beta2 possibly regulates the differentiation of pulp cell at specific stages synergistically with other factors.

Introduction: This study aims to assess the accuracy of detection of breast lesion by breast self-examination and to assess different factors affecting the accuracy., Methods: All consecutive Chinese female patients, who attended our breast imaging unit in 2001, completed our questionnaire, had retrievable hard copy films, and had more than three years clinical follow-up, were recruited for this study. Different factors, such as age, menopausal status, previous experience of breastfeeding, family history of breast cancer, previous history of mastectomy or lumpectomy, hormonal therapy, oral contraceptive pills and previous history of mammography, were correlated with accuracy in self-detection of breast lesions retrospectively. The nature, size and location of the lesion, and breast size based on imaging, were also correlated with the accuracy in self-detection of breast lesions., Results: A total of 163 questionnaires were analysed. 111 patients detected a breast lesion themselves and 24 of these lesions were false-positives. A total of 173 lesions (27 cancerous, 146 benign lesions) were documented by either ultrasonography and/or mammography, and confirmed by either histology or three-year clinical follow-up. The overall sensitivity in detecting both benign and malignant breast lesions was 71% when number of breast lesions was used as the denominator, and up to 78% sensitivity was achieved when number of patients was used as the denominator. History of mastectomy, and size and nature of the lesions were found to affect the accuracy of self-detection of breast lesions., Conclusion: Overall, breast self-examinations were effective in the detection of breast lesions and factors such as size of lesion, nature of the lesion and history of mastectomy affect the accuracy of the detections. Breast self-examination should be promoted for early detection of breast cancer.

This article takes a special focus on signal amplification technologies in immunoassays and new generations of lateral-flow assays. Novel signal amplification technologies based either on new classes of biofunctional nanocrystals consisting of releasable fluorophores or on aggregation-induced emission (AIE) can improve the sensitivity and the limits of detection in immunoassays. A bio-barcode assay also allows signal amplification by utilizing antibody-coated magnetic beads to concentrate the analytes and antibody-coated gold nanoparticle probes to carry with a large number of oligonucleotides. These innovative technologies boost the development of immunoassays. Growth in rapid immunoassay is fueled by the increasing number of diabetics, the globalization of infectious diseases and the surge in cardiovascular and other chronic diseases as well as other chronic conditions. Rapid, near patient, decentralized, point-of-care (POC) tests are emerging as a tool for more efficient diagnosis and patient evaluation. Technological innovations in lateral-flow assays have enabled a move to bring testing closer to the patient. A novel "digital-style" lateral-flow assay provides semi-quantitative results by simply counting the number of red lines in the test without any expensive reading instrument. An immuno-threshold-based assay can give a signal directly proportional to the concentration of a hapten to prevent confusion on interpretation of the test results. In addition, POC tests become more meaningful to healthcare professionals by combining the benefits of new technologies to provide quantitative results. A molecular compact disc provides a high-resolution imaging capability that can identify and quantify many different antigens simultaneously in highly complex immunoassays. Further advances in immunoassays will bring diagnostic testing even closer to the patient, and can help physicians to monitor diseases that require immediate test results, thereby enhancing the quality of patient care.

Aim: To evaluate the antioxidant and pro-oxidant properties of chlorhexidine (CHX)., Methodology: The scavenging and generation of reactive oxygen species (ROS) by CHX in the presence or absence of saturated Ca(OH)(2) solutions was evaluated. The reaction emitted chemiluminescence in the presence of lucigenin thus was determined by a luminometer to evaluate the levels of ROS production. Changes in DNA conformation were analysed by agarose gel electrophoresis. Paired Student's t-test was used to compare the difference between groups., Results: Chlorhexidine (0.00002-0.02%) effectively scavenged 56-88% of the superoxide radicals generated by the xanthine/xanthine oxidase reaction. Through analysis of PUC18 DNA conformation changes, CHX was shown to be a mild scavenger of hydroxyl radicals generated by H(2)O(2) plus FeCl(2). However, CHX (>0.083%) decreased the mobility of PUC18 plasmid DNA with potential production of DNA-DNA cross-link and severe DNA breaks (presence of DNA smear) at further higher concentrations. Furthermore, CHX induced ROS production including H(2)O(2) and superoxide radicals in 0.1N NaOH (pH = 12.76) or Ca(OH)(2) (pH = 12.5) solutions., Conclusion: Chlorhexidine exhibited both antioxidant and pro-oxidant properties under different conditions. These events are possibly involved in the killing of root canal and periodontal microorganisms when CHX and Ca(OH)(2) were used in combination or separately. Potential genotoxicity and tissue damage when extruded into the periradicular tissue and at higher concentrations should be considered during periodontal and endodontic practice.

Aim: Surgical and nonsurgical procedures for management of hepatolithiasis have been reported. The aim of this study was to evaluate immediate and long-term results of hepatectomy as treatment for hepatolithiasis., Materials and Methods: Immediate and long-term outcomes of 123 consecutive patients who underwent hepatectomy for hepatolithiasis at our institution from 2000 to 2005 were analyzed retrospectively. Acute cholangitis was the major presenting symptom (in 106 out of 123, 86.2% of cases)., Results: The immediate stone clearance rate was 92.7% (114 out of 123) and final stone clearance rate was 96% (118 out of 123) after subsequent T-tube route or endoscopic retrograde cholangiopancreatography (ERCP). Residual stones were identified in 5 patients (4%). The surgical morbidity and mortality rates were 33.3% (41 out of 123) and 1.6% (2 out of 123) respectively. Of the 123 patients, 3 (2.4%) had associated cholangiocarcinoma at the time of hepatectomy. With a median follow-up of 40.3 months (range 5-58), a recurrent stone developed in 7 patients (5.7%) and cholangiocarcinoma in 2 (1.6%). Ten patients died during the follow-up period, with 4 of them (out of 123, 3.2%) due to recurrent stone with sepsis., Conclusion: Hepatectomy is a safe and effective treatment for hepatolithiasis, with a high stone clearance rate and fair rate of surgical complications. Recurrent stone-induced sepsis and cholangiocarcinoma are the major factors compromising long-term survival in these patients.

Advances in nanotechnology have had significant impacts in the field of biodiagnostics. In this study, we describe the novel application of dissolvable, organic and biofunctional nanocrystals for the quantitative detection of a PCR product. Fluorescein diacetate (FDA), a fluorogenic precursor of fluorescein, was milled in a solution of a polymeric surfactant to create a stable, nanosized colloid with an interface for coupling streptavidin molecules. The application of these particulate labels for the quantitative detection of biotinylated human papillomavirus (HPV) DNA, amplified in a standard PCR procedure, was demonstrated. After the affinity reaction, the FDA molecules were dissolved and concomitantly converted into fluorescein. This approach resulted in a high selectivity, short incubation times and a sensitivity up to 147 times greater than obtained from state-of-the-art, directly fluorescent-labeled streptavidins. This innovative method offers rapid detection of small amounts of nucleic acids because less target material and thus fewer PCR cycles are required.

Sanguinarine is a benzopheanthridine alkaloid present in the root of Sanguinaria canadensis L. and Chellidonium majus L. In this study, sanguinarine (2 and 3 microM) exhibited cytotoxicity to KB cancer cells by decreasing MTT reduction to 83% and 52% of control after 24-h of exposure. Sanguinarine also inhibited the colony forming capacity (>52-58%) and growth of KB cancer cells at concentrations higher than 0.5-1 microM. Short-term exposure to sanguinarine (>0.5 microM) effectively suppressed the adhesion of KB cells to collagen and fibronectin (FN). Sanguinarine (2 and 3 microM) induced evident apoptosis as indicated by an increase in sub-G0/G1 populations, which was detected after 6-h of exposure. Only a slight increase in cells arresting in S-phase and G2/M was noted. Induction of KB cell apoptosis and necrosis by sanguinarine (2 and 3 microM) was further confirmed by Annexin V-PI dual staining flow cytometry and the presence of DNA fragmentation. The cytotoxicity by sanguinarine was accompanied by an increase in production of reactive oxygen species (ROS) and depolarization of mitochondrial membrane potential as indicated by single cell flow cytometric analysis of DCF and rhodamine fluorescence. NAC (1 and 3 mM) and catalase (2000 U/ml) prevented the sanguinarine-induced ROS production and cytotoxicity, whereas dimethylthiourea (DMT) showed no marked preventive effect. These results suggest that sanguinarine has anticarcinogenic properties with induction of ROS production and mitochondrial membrane depolarization, which mediate cancer cell death.

Perturbation of the function of endoplasmic reticulum (ER) causes stress leading to the activation of cell signaling pathways known as the unfolded protein response (UPR). Severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) uses ER as a site for synthesis and processing of viral proteins. In this report, we demonstrate that infection with SARS-CoV induces the UPR in cultured cells. A comparison with M, E, and NSP6 proteins indicates that SARS-CoV spike (S) protein sufficiently induces transcriptional activation of several UPR effectors, including glucose-regulated protein 78 (GRP78), GRP94, and C/EBP homologous protein. A substantial amount of S protein accumulates in the ER. The expression of S protein exerts different effects on the three major signaling pathways of the UPR. Particularly, it induces GRP78/94 through PKR-like ER kinase but has no influence on activating transcription factor 6 or X box-binding protein 1. Taken together, our findings suggest that SARS-CoV S protein specifically modulates the UPR to facilitate viral replication.

Objective: Neopterin is generated and released in increased amounts by macrophages upon activation by interferon-gamma during Th1-type immune response. The potential usefulness of neopterin in early prognostic information of dengue virus infection was investigated., Methods: Neopterin concentrations were determined in serum samples from 110 dengue fever (DF) patients. The neopterin levels were compared with those in 50 measles and 40 influenza patients; 155 healthy blood donors served as controls., Results: In acute sera of DF patients mean neopterin concentration was 48.2 nmol/L, which was higher than that in patients with measles (mean: 36.3 nmol/L) and influenza (18.8 nmol/L) and in healthy controls (6.7 nmol/L; P<0.001). In the patients with confirmed DF, an early neopterin elevation was detected already at the first day after the onset of symptoms and rose to a maximum level of 54.3 nmol/L 4 days after the onset. Higher increase of neopterin level in DF patients was associated with longer duration of fever and thus predicted the clinical course of the disease., Conclusions: Neopterin concentrations were found significantly higher in DF patients compared with healthy controls and also with other viral infections (P<0.001) and may allow early assessment of the severity of DF.

A solid-phase sandwich fluorescence immunoassay using nanocrystals of a fluorogenic precursor, fluorescein diacetate (FDA), conjugated with monoclonal antibodies for the detection of C-reactive protein (CRP), is described. FDA nanocrystals were coated with distearoylglycerophosphoethanolamine (DSPE), modified with amino(poly(ethylene glycol))(PEG(2000)-Amine) as an interface for coupling biomolecules. CRP was chosen as a model analyte because of its widely accepted role as a marker for acute inflammation and prospective heart failure. A low limit of detection (1.10 microg l(-1)) and high precision (CV = 2.72-9.48%) were achieved. Following the immunoreaction, the monoclonal anti-CRP conjugated nanocrystals were released by hydrolysis and dissolution instigated by the addition of a large volume of organic solvent-sodium hydroxide mixture. Using human serum samples from 66 patients with high heart attack risk and 19 healthy blood donors, this CRP fluorescence immunoassay showed a good correlation to the commercially available, turbidimetric immunoassay for CRP. This result was corroborated by the Bland-Altman plot that showed a mean difference between the two methods of only 0.36+/-1.46 mg l(-1). The study demonstrates that the organic fluorogenic FDA nanocrystals can be applied for the detection of CRP, which is a clinically interesting plasma protein with a low limit of detection.

Background: Chronic infection and inflammation, including periodontitis, is linked to an increased risk for atherosclerosis. To investigate the possible adverse effects of periodontitis in maintenance hemodialysis patients, we compared periodontal severity with malnutrition and inflammation, which are associated with poor atherosclerotic outcome in hemodialysis patients., Methods: Two hundred fifty-three hemodialysis patients were included in this study to evaluate clinical periodontal status by using the Plaque Index, Gingival Index, and Periodontal Disease Index. Geographic, hematologic, biochemical, and dialysis-related data also were collected. Values for nutritional and inflammatory markers, such as albumin, blood urea nitrogen, creatinine, transferrin, absolute lymphocyte count, normalized protein catabolic rate, high-sensitivity C-reactive protein, and ferritin, were included for analysis with the Periodontal Index., Results: Poor oral health status was shown by 80.6% of hemodialysis patients with periodontal disease. In an analysis of geographic and disease-related parameters, we found that aging, smoking, diabetes, and longer dialysis duration were associated with severity of periodontitis. Parameters of malnutrition and inflammation also were associated with poor periodontal status. We next conducted multiple regression analysis and found that age, diabetes, smoking, albumin level, and dialysis duration were associated independently with periodontitis severity in hemodialysis patients. According to the severity of periodontitis, there were higher percentiles of patients with malnutrition (chi-square = 13.055; P = 0.005) and inflammation (chi-square = 10.046; P = 0.018) in the severe group., Conclusion: Periodontal health is poor in hemodialysis patients and correlates with markers of malnutrition and inflammation. Its diagnosis and treatment deserve better awareness.

Nanocrystals of the fluorogenic precursor fluorescein diacetate (FDA) were applied as labels in order to improve on the assay sensitivity achieved in our previous studies. Each FDA nanocrystal can be converted into approximately 2.6x10(6) fluorescein molecules, which is useful for improving immunoassay sensitivity and limits of detection. NeutrAvidin was simply adsorbed onto the surface of the FDA nanocrystals, which were coated with distearoylglycerophosphoethanolamine (DSPE) modified with amino(poly(ethylene glycol))(PEG(2000)-Amine) as an interface for coupling biomolecules. This can be applied to detect different kinds of analytes that are captured by corresponding biotinylated biomolecules in different bioanalytical applications. The applicability of the NeutrAvidin-labeled nanocrystals was demonstrated in an immunoassay using the labeled avidin-biotin technique. Biotinylated antibody and FDA-labeled avidin were applied to the assay sequentially. The performance was compared with the traditional sandwich-type assay for mouse immunoglobulin G detection. Following the immunoreaction, the nanocrystals were released by hydrolysis and dissolution instigated by adding a large volume of organic solvent/sodium hydroxide mixture. The limit of detection was lower (by a factor of 2.5-21) and the sensitivity was (3.5-30-fold) higher than immunoassays using commercial labeling systems (FITC and peroxidase). This study shows that using fluorescent nanocrystals in combination with the avidin-biotin technique can enhance assay sensitivity and provide a lower limit of detection without requiring long incubation times as in enzyme-based labels.

1.--Thrombin is activated during gingival tissue injury and inflammation. Thrombin (platelet)-rich plasma has been used for periodontal regeneration with success. Thrombin and other bacterial proteases also affect the functions of adjacent periodontal cells via stimulation of protease-activated receptors (PARs). 2.--We noted that thrombin (0.1-2 U ml(-1)), human, and frog PAR-1 agonist peptide (20-240 microM) induced the gingival fibroblast (GF)-populated collagen gel contraction within 2 h of exposure. However, PAR-2, PAR-3, and PAR-4 agonist peptide (20-240 microM) showed little effect on collagen gel contraction. U73122 (phospholipase C inhibitor) and 2-APB (IP3 antagonist) were effective in inhibition of GF contraction. 3.--Thrombin-induced GF contraction was inhibited by 5 mM EGTA (an extracellular calcium chelator) and verapamil (an L-type calcium channel blocker). In addition, W7 (10 and 25 microM, a calcium/calmodulin (CaM) inhibitor), ML-7 (50 microM, myosin light chain kinase (MLCK) inhibitor), and HA1077 (100 microM, Rho kinase inhibitor) completely inhibited the thrombin-induced collagen gel contraction. Thrombin also induced the phosphorylation of ERK1/ERK2 and elevated the Rho-GTP levels in GF. 4.--However, U0126 only partially inhibited the thrombin-induced GF contraction. Similarly, wortmannin (100 nM), LY294002 (20 microM) (two PI3K inhibitor) and genistein also showed partial inhibition. Moreover, NAC was not able to suppress the GF contraction, as supported by the slight decrease in reactive oxygen species production in GF by thrombin. 5.--Thrombin also stimulated metalloproteinase-2 (MMP-2) and MMP-3 production in GF. But addition of GM6001 or 1,10-phenanthroline, two MMP inhibitors, could not inhibit the thrombin-induced GF contraction. 6.--These results indicate that thrombin is crucial in the periodontal inflammation and wound healing by promoting GF contraction. This event is mainly mediated via PAR-1 activation, PLC activation, extracellular calcium influx via L-type calcium channel, and the calcium/CaM-MLCK and Rho kinase activation pathway.

Despite recent advances in management of severe blunt liver trauma, the operative mortality rate in patients with complicated blunt liver trauma (grades IV and V) is still high. The purpose of this study was to assess the results of anatomic liver resection for severe liver trauma in the institution in which liver transplant surgeons are responsible for the management of liver injuries.

Neopterin and C-reactive protein (CRP) concentrations were determined in serum samples from 129 severe acute respiratory syndrome (SARS) patients and 156 healthy blood donors. In the patients with confirmed SARS, an early neopterin elevation was detected already at the day of onset of symptoms and rose to a maximum level of 45.0 nmol/L 3 days after the onset. All SARS patients had elevated neopterin concentrations (>10 nmol/L) within 9 days after the onset. The mean neopterin concentrations were 34.2 nmol/L in acute sera of SARS patients, 5.1 nmol/L in convalescent sera, and 6.7 nmol/L in healthy controls. In contrast, the mean CRP concentrations in both acute and convalescent sera of SARS patients were in the normal range (<10 mg/L). Serum neopterin level in SARS patients was associated with fever period and thus the clinical progression of the disease, while there was no significant correlation between the CRP level and the fever period. Serum neopterin may allow early assessment of the severity of SARS. The decrease of neopterin level was found after steroid treatment, which indicates that blood samples should be collected before steroid treatment for the neopterin measurement.

Fatty acid-binding protein (FABP) holds promise for early detection of tissue injury. This small protein (15kD) appears earlier in the blood than large proteins after cell damage. Combined its characteristics of high concentration tissue contents and low normal plasma values provide the possibility of a rapid rise above the respective reference values, and thus an early indication of the appearance of tissue injury. A general review was presented on the current status of different types of FABP for the detection of tissue injury in patients with myocardial injury, brain injury and also in athletes or horses with skeletal muscle injury. To take full advantage of the characteristics of the early marker FABP, rapid analysis is a crucial parameter. In this review, an overview of the development of immunoassay for the quantification of FABP in buffer, plasma or whole blood was outlined. The characteristics of different FABP immunosensors and immunotests were described. The feasibility of these immunoassays to be used in routine clinical practice and in emergency case was also discussed. Nowadays, the improved automated immunoassays (e.g. a microparticle-enhanced turbidimetric immunoassay), less time-consuming bedside immunosensors and immunotests (e.g. a one-step FABP lateral flow immunotest), are the main advance technology in point-of-care testing. With these point-of-care tests, the application of FABP as an early tissue injury marker has a great potential for many clinical purposes.

Transforming growth factor-beta (TGF-beta) is important in regulating the repair and regeneration of damaged dental pulp. For further elucidating the roles of different isoforms of TGF-beta in the healing and inflammatory processes of human dental pulp, we found that TGF-beta1, TGF-beta2 and TGF-beta3 inhibited the growth of two human dental pulp cell strains in vitro by 19-29, 18-25 and 23-26%, respectively, at a concentration of 0.5 ng/ml. TGF-beta also differentially stimulated the collagen synthesis of pulp cells. Collagen synthesis increased by 1 ng/ml of TGF-beta1 and TGF-beta2 by 42 and 51%, respectively. TGF-beta3 (0.1-1 ng/ml) lacked of stimulatory effect on collagen synthesis of pulp cells. Pulp cells have the intrinsic capacity to contract collagen lattice, leading to decreasing of lattice diameter. An 8 h exposure to TGF-beta1 and TGF-beta2 enhanced the pulp cell-populated collagen lattice contraction at concentrations ranging from 0.2 to 3 ng/ml. At similar concentrations, TGF-beta3 lacked of this stimulatory effect. When collagen lattice were detached after 24 h of exposure, TGF-beta1 and TGF-beta2 (0.6-3 ng/ml) induced the pulp cells-populated collagen lattice contraction within 4-8h of gel detachment. These results indicate that TGF-beta-induced collagen lattice contraction is a late cellular event. These in vitro results indicate that effects of TGF-beta isoforms on the growth, collagen synthesis and collagen lattice contraction of pulp cells may play crucial roles in the pathobiological processes of dental pulp.

The compaction of DNA by a cationic surfactant both in the bulk and adsorbed on the surface of latex particles was followed for the first time by SANS. In the bulk, a decrease in the overall size of the DNA coil in the presence of the cationic surfactant was observed at a negative-to-positive charge ratio far below the phase separation region, at a negative-to-positive charge ratio of 18. Additionally, large surfactant aggregates seem to form within the DNA-surfactant complex. On the other hand, DNA adsorbs onto the surface of latex particles, forming a thick layer, as evidenced by the fitting of the SANS data to a core-shell form factor. Addition of a cationic surfactant to the DNA-coated latex particles at a negative-to-positive charge ratio of 38 induces a slight decrease in the size of the particle layer, where the cationic surfactant is evenly distributed within the adsorbed layer. A further decrease of the negative-to-positive charge ratio to 18 induces a dramatic change in the SANS data that suggests significant compaction of the adsorbed layer and the formation of large surfactant aggregates, similar to those detected in the bulk.

We have previously characterized transcription factor LZIP to be a growth suppressor targeted by hepatitis C virus oncoprotein. In search of proteins closely related to LZIP, we have identified a liver-enriched transcription factor CREB-H. LZIP and CREB-H represent a new subfamily of bZIP factors. CREB-H activates transcription by binding to cAMP responsive element, box B, and ATF6-binding element. Interestingly, CREB-H has a putative transmembrane (TM) domain and it localizes ambiently to the endoplasmic reticulum. Proteolytic cleavage that removes the TM domain leads to nuclear translocation and activation of CREB-H. CREB-H activates the promoter of hepatic gluconeogenic enzyme phosphoenolpyruvate carboxykinase. This activation can be further stimulated by cAMP and protein kinase A. CREB-H transcript is exclusively abundant in adult liver. In contrast, the expression of CREB-H mRNA is aberrantly reduced in hepatoma tissues and cells. The enforced expression of CREB-H suppresses the proliferation of cultured hepatoma cells. Taken together, our findings suggest that the liver-enriched bZIP transcription factor CREB-H is a growth suppressor that plays a role in hepatic physiology and pathology.

Cresol is a well-known environmental pollutant, toluene metabolite, uremic toxicant and accidental poisoning product. Formocresol, a preparation of formalin and cresol, is also used as a root canal medicament and for pulpotomy of primary teeth. However, little is known about its effect on cardiovascular system. In this study, m-cresol inhibited the AA-induced platelet aggregation by 43-97% at concentrations ranging from 0.25 to 1 mM. Collagen-induced platelet aggregation was also inhibited by 0.25-1 mM of m-cresol by 47-98%. Accordingly, o-cresol (0.1-0.5 mM) also inhibited the AA-induced platelet aggregation by 46-96% and the collagen-induced platelet aggregation by 35-88% at concentrations of 0.1-1 mM. AA- and collagen-induced platelet thromboxane B(2) (TXB(2)) production was inhibited by even 0.1 mM of m-cresol with 88 and 54% of inhibition, respectively. The o-cresol (0.1 mM) also inhibited the AA- and collagen-induced platelet TXB(2) production with 91 and 97% respectively. Although m- and o-cresol (<1 mM) showed little effect on thrombin-induced platelet aggregation, they effectively inhibited the thrombin-induced platelet TXB(2) production. The m-cresol (2 and 5 mM) inhibited the COX-1 activity by 55-99%, but showed little effect on COX-2 enzyme activity. Moreover, o-cresol (0.5 and 1 mM) inhibited the COX-1 activity by 40-95%. COX-2 enzyme activity was inhibited by 68% at a concentration of 5 mM o-cresol. These results indicate that acute cresol-poisoning, direct root canal medication with formocresol or long-term occupational exposure to cresol and toluene may potentially suppress blood clot formation and lead to tissue hemorrhage via inhibition of platelet aggregation, TXB(2) production and COX enzyme activity.

A novel class of biofunctional silole nanocrystals with the potential to create highly sensitive immunoassay was firstly demonstrated. Biolabels were constructed by encapsulating nanocrystalline hexaphenylsilole [Ph2Si(CPh)4HPS] within ultrathin polyelectrolyte layers via the layer-by-layer (LbL) technique that provided an "interface" for the attachment of antibodies. A high ratio of fluorescent dyes to biomolecules (F/P ratio; 2.4 x 10(3)) was achieved without self-quenching problem. The aggregation-induced emission (AIE) feature offered silole biolabels the sensitivity 40- to 140-fold higher than that of a start-of-the-art immunoassay using directly fluorescent-labeled antibodies.

Recently, numerous attempts have been made to improve the performance of fluorescence immunoassays. One way pursued is the substitution of labeling molecules by micro- or nanocrystalline dyes. The surfaces of these particulate structures are typically engineered by a layerwise assembly of oppositely charged polyelectrolytes, the outer layer being constituted of biorecognition molecules, for example, immunoglobulins. In this study, we show that amphiphilic polymers such as alkylated poly(ethylene imine)s and 1,2-distearoyl-sn-glycero-3-phosphatoethanolamine-N-[amino(poly(ethylene glycol))] can fully substitute the more intricate layer-by-layer technique and evaluate the influence of surface charge and particle size on the overall performance of these assays., (Copyright 2004 American Chemical Society)

A novel signal amplification technology based on a new class of biofunctional fluorescent nanocrystals holds promise to improve the sensitivity and the limits of detection of immunoassays. A two-step approach without layer-by-layer techniques is described to encapsulate the fluorogenic precursor fluorescein diacetate (FDA) nanocrystals (107-nm average size) followed by conjugation of the antibody. Distearoylphosphatidylethanolamine (DSPE) modified with amino(poly(ethylene glycol)) (PEG(2000)Amine) is coated on the surface of the FDA nanocrystals to provide a interface for the antibody coupling. Anti-mouse antibodies are attached to the nanocrystalline FDA biolabels by adsorption. A high molar ratio of fluorescent molecules to biomolecules (2.8 x 10(4)) is achieved in this nanocrystal biolabel system. The analytical performance of the nanocrystal-based label system is evaluated in a model sandwich immunoassay for the detection of mouse IgG. After separation of the nonbound antibody nanocrystal labels, fluorophores are released by hydrolysis and dissolution of the nanocrystalline FDA. Due to the release of the fluorophores (fluoresceins) into a large volume of organic solvent/sodium hydroxide mixture, self-quenching is suppressed. The FDA[DSPE-PEG(2000)Amine]-modified biolabels have a highly stable colloidal suspension with minimized nonspecific interactions. The limit of detection was lowered by a factor of 5-28, and the sensitivity was 400-2700-fold higher compared with a state-of-the-art immunoassay using directly fluorescent-labeled antibodies. Our approach provides high sensitivity and low limits of detection without the need for long incubation times, making it an interesting alternative in biolabel technology.

Thrombin is a serine protease activated during injury and inflammation. Thrombin and other proteases generated by periodontal pathogens affect the behavior of periodontal cells via activation of protease-activated receptors (PARs). We noted that thrombin and PAR-1 agonist peptide stimulated intracellular calcium levels ([Ca2+]i) of gingival fibroblasts (GF). This increase of [Ca2+]i was inhibited by EGTA and verapamil. U73122 and neomycin inhibited thrombin- and PAR-1-induced [Ca2+]i. Furthermore, 2-APB (75-100 microM, inositol triphosphate [IP3] receptor antagonist), thapsigargin (1 microM), SKF-96365 (200 microM) and W7 (50 and 100 microM) also suppressed the PAR-1- and thrombin-induced [Ca2+]i. However, H7 (100, 200 microM) and ryanodine showed little effects. Blocking Ca2+ efflux from mitochondria by CGP37157 (50, 100 microM) inhibited both thrombin- and PAR-1-induced [Ca2+]i. Thrombin induced the IP3 production of GF within 30-seconds of exposure, which was inhibited by U73122. These results indicate that mitochondrial calcium efflux and calcium-calmodulin pathways are related to thrombin and PAR-1 induced [Ca2+]i in GF. Thrombin-induced [Ca2+]i of GF is mainly due to PAR-1 activation, extracellular calcium influx via L-type calcium channel, PLC activation, then IP3 binding to IP3 receptor in sarcoplasmic reticulum, which leads to intracellular calcium release and subsequently alters cell membrane capacitative calcium entry.

Human heart-type fatty acid-binding protein (FABP) has a high potential as an early marker for myocardial infarction (MI) being more specific than myoglobin. FABP is a low molecular mass cytoplasmic protein (15 kDa) that is released early after the onset of ischemia and it may be useful for rapid confirmation or exclusion of acute myocardial infarction (AMI). Immunochemically assayed FABP, cardiac troponin I (cTnI) and enzymatically assayed creatine phosphokinase (CPK) were determined serially in plasma and serum samples from 218 patients presenting with chest pain and suspected MI. In the 94 patients with confirmed MI, FABP rose to a maximum level (577.6 +/- 43.8 microg/L) 3 hours after the onset of symptoms and returned to normal within 30 hours. The FABP level peaked 7-9 hours earlier than CPK (2288 +/- 131 U/L) and cTnI (357.1 +/- 23.9 microg/L). CPK took 50-70 hours to return to normal level and cTnI returned to normal level over 70 hours. The areas under the receiver operating characteristic (ROC) curves for FABP were calculated as 0.871 at admission and 0.995 one hour after admission, whereas for CPK the areas were 0.711 and 0.856 and for cTnI the areas were 0.677 and 0.845, indicating that the FABP test gave a better diagnostic classification at the early stage being reached by cTnI (0.995) only 8 hours after admission. For FABP, both sensitivity and negative predictive value (NPV) increased quickly to 100% for samples monitored just one hour after admission. By using only two samples, one at admission and one 1 hour post admission, sequential FABP monitoring can reliably diagnose AMI patients 1 hour after admission and 100% of non-AMI patients can be excluded with no false negative results. The late markers cTnI and CPK have the similar diagnostic performance only 7 hours later. Thus measurement of FABP in plasma or serum allows the earliest immunochemical confirmation or exclusion of AMI.

Thirteen patients with large ameloblastomas of the mandible underwent segmental mandibulectomy and immediate reconstruction, with simultaneous placement of osseointegrated implants. All patients received palatal mucosal grafts around the dental implants 6 to 10 months after surgical treatment and received implant-supported prostheses another 1 to 2 months later. There were five female and eight male patients, with a mean age of 32 years (range, 17 to 50 years). The mean length of the mandibular defect was 8.8 cm (range, 5 to 13 cm). All free fibula flap procedures were successful, with no reexplorations or partial flap losses. There was no clinical or radiographic evidence of failure during the osseointegration process for any implant. With functional occlusal loading, the marginal bone loss around the implants was less than 1.5 mm in a mean follow-up period of 40 months (range, 18 to 70 months). There were no recurrences during that time. The technique described allows improved access to the bone at the time of reconstruction, immediate assessment of alveolar ridge relationships, and accurate fixation of the implant-fibula construct. The advantages of this procedure include a reduced risk of recurrence with segmental resection, reliable mandibular reconstruction, and reduction of the number of surgical procedures, allowing full oral rehabilitation in a shorter time. It is concluded that segmental mandibulectomy and immediate vascularized fibula osteoseptocutaneous flap reconstruction, with simultaneous placement of osseointegrated implants, represent an ideal treatment method for large ameloblastomas of the mandible.

The major challenges for anti-hepatitis B Virus (HBV) therapy are the low efficacy of current drugs and the occurrence of drug resistant HBV mutations. A drug with new target sites or independent metabolic pathways may overcome these shortcomings. Small interfering RNA (siRNA) offers the possibility of developing a new anti-HBV therapy. Here we describe the almost complete inhibition of HBV replication by stably expressed 21-mer short hairpin RNAs (shRNA). Besides the conventional targets on HBV reverse-transcriptase, we also systemically targeted other sites of pregenomic RNA, including direct repeat (DR) elements, S, core, and X gene. Our results indicated that shRNAs can serve as efficient alternative anti-HBV agents. They can also be used in combination with chemotherapy, because they showed better effects on the inhibition of HBV replication due to different mechanisms of drug actions.

Background: Loss of the interproximal dental papilla may cause functional and, especially in the maxillary anterior region, phonetic and severe esthetic problems. The purpose of this study was to investigate whether the distance from the contact point to the bone crest on standardized periapical radiographs of the maxillary anterior teeth could be correlated with the presence of the interproximal papilla in Taiwanese patients., Methods: In total, 200 interproximal sites of maxillary anterior teeth in 45 randomly selected patients were examined. Selected subjects were adult Taiwanese with fully erupted permanent dentition. The presence of the interproximal papilla was determined visually. If there was no visible space apical to the contact area, the papilla was recorded as being present. The distance from the contact point to the crest of bone was measured on standardized periapical radiographs using a paralleling technique with a RinnXCP holder., Results: Data revealed that when the distance from the contact point to the bone crest on standardized periapical radiographs was 5 mm or less, the papillae were almost 100% present. When the distance was 6 mm, 51% of the papillae were present, and when the distance was 7 mm or greater, only 23% of the papillae were present., Conclusion: The distance from the contact point to the bone crest on standardized periapical radiographs of the maxillary anterior teeth is highly associated with the presence or absence of the interproximal papilla in Taiwanese patients, and is a useful guide for clinical evaluation.

Betel quid (BQ) chewing is an etiologic factor of oral cancer and submucus fibrosis (OSF). Keratinocyte inflammation is crucial for the pathogenesis of cancer and tissue fibrosis. We found that areca nut (AN) extract (100-400 micro g/ml) induced PGE2 production by KB cells by 2.34- to 23.1-fold and also TNF-alpha production by gingival keratinocytes (GK). Arecoline (0.2-1.2 mM) elevated PGE2 production by KB cells by 2.5- to 6.1-fold. AN extract (200-400 micro g/ml) also induced IL-6 production by GK (7.5- to 8.4-fold) and KB cells. In contrast, arecoline (0.1-1.2 mM) suppressed IL-6 production by GK and KB cells, with 42-81 and 41-63% inhibition, respectively. A 48 h exposure of GK to 800-1200 micro g/ml AN extract led to 37-69% cell death. Arecoline cytotoxicity to GK was noted at concentrations of 0.8-1.2 mM, which led to 28-38% cell death. AN extract (400-800 micro g/ml) induced Cox-2 and IL-6 mRNA expression and also COX-2 protein production by KB cells. IL-6 (5-100 ng/ml) suppressed GK growth by 20-33%, but enhanced oral fibroblast (OMF) and KB cell growth. PGE2 (0.05-5 micro g/ml) and anti-IL-6 antibody (ab) (50-1000 ng/ml) showed little effect on GK and KB cell growth. Incubation of GK and KB cells with aspirin, anti-IL-6 ab and anti-TNF-alpha ab showed little effect on arecoline- and AN-induced cytotoxicity, cell cycle arrest and apoptosis. Exposure to anti-TNF-alpha ab slightly affected arecoline- and AN-modulated PGE2 and IL-6 production by GK and KB cells. Arecoline- and AN-conditioned medium decreased phytohemagglutinin-mediated CD4+ and CD8+ T cell activation. These results indicate that BQ chewing contributes to the pathogenesis of cancer and OSF by impairing T cell activation and by induction of PGE2, TNF-alpha and IL-6 production, which affect oral mucosal inflammation and growth of OMF and oral epithelial cells.

Using human heart-type fatty acid-binding protein (H-FABP) as an early cardiac marker to confirm or exclude a diagnosis of acute myocardial infarction (AMI) soon after admission requires a rapid assay system. Due to the requirement of skillful technicians and complicated assay procedures, most immunochemical assays for H-FABP are of limited use for routine clinical practice. In the present study, we describe a rapid lateral-flow assay for detection of H-FABP. Fifty-one human samples were evaluated using a conventional ELISA and a newly developed lateral-flow assay. A good agreement between the two methods was found according to Bland and Altman plot. The correlation found was y=0.9685 x -0.6270 (r(2)=0.9585). The detector antibody labeled with colloidal gold was mixed with those without label to extend the linear range of the calibration curve up to 125 microg/l H-FABP with r(2)=0.9832. The detection limit of the assay was 2.8 microg/l. The test-strips can be stored either at 4 degrees C and room temperature for up to 1 year without significant loss of activity. Finally, a one-step FABP test so-called CardioDetect(R), which was derived from the serum lateral-flow assay has been designed for qualitative determination of H-FABP in whole blood samples. It requires no sample pretreatment and gives results within 15 min. Thirty-eight patients presenting with chest pain and suspected AMI were studied. Using an upper reference level of 7 microg/l, the specificity of the rapid test was 94%. Both sensitivity and negative predictive value (NPV) were 100%, implying that 100% of non-AMI patients could be excluded with no false-negative results. With this rapid and sensitive immunotest, H-FABP could soon be introduced into clinical practice.

Unsuspected gallbladder cancer after laparoscopic cholecystectomy (LC) is a cancer that was previously manipulated by laparoscopic technique. The reported incidence was 0.3-1% and became an emerging problem as the popularity of LC increased. Lack of reliable data could address the outcome of reresection or nonreresection patients and the standard management. This study reviewed a single center experience in managing unsuspected gallbladder cancer patients after LC between July 1, 1992 and July 1, 2000 who had at least 2 years of follow-up. There were 11 patients (0.6%) postoperatively diagnosed with gallbladder cancer after 1825 LCs. Group A included three patients (28%) with nontransmural invasion, group B included four patients (36%) who had transmural invasion without secondary surgical intervention, and group C included four patients (36%) with reresection. The perioperation parameters and strategies were collected and compared. A review of the literature was performed simultaneously, and we concluded that unsuspected gallbladder cancer with nontransmural invasion needs no further treatment; however, aggressive reresection is beneficial to transmural invasion cancer, and prevention of bile spillage during LC should be the goal of every surgeon.

Because of the efforts of many pioneer surgeons, the minimally invasive video-assisted thyroidectomy (MIVAT) has been recognized as a safe procedure, offering advantages such as better cosmetic outcome and less analgesic need. The MIVAT technique was described in 51 selected patients in 2001. The technique was not therefore widely used because of the excess operating time compared with traditional thyroidectomy, and most importantly, this method needed a steep learning period. This study reports a modified MIVAT procedure, which can make this operation easier and shorten the time of learning. We compared the outcomes of the originally described methods with our modified method. The selection criteria for performing MIVAT were as follows: thyroid nodules in one lobe and less than 50 mm on their largest diameter, benign lesion proved by fine-needle biopsy, patient without history of thyroiditis, and no previous neck surgery or irradiation. All patients received lobectomy. Sixty patients were eligible for MIVAT during a period of 27 months. The patients were divided into two groups. Group A consisted of the 17 patients who underwent MIVAT using the original technique that was described previously. Group B consisted of the 43 patients who underwent MIVAT using a self-designed Army retractor with a mosaic ring. The mean operation time of Group A was 120 minutes and that of Group B was 59.2 minutes. The size of the incisions was no difference in either group. There were no postoperative complications except in one patient with transient recurrent laryngeal nerve palsy in Group A. There was one conversion to open thyroidectomy in Group A and none in Group B. The cosmetic results were no different between the two groups. In conclusion, the use of a modified Army retractor with a mosaic ring made the MIVAT procedure easier and offered similar advantages.

Piper betle, belonging to the Piperaceae family, is a tropical plant, and its leaf and inflorescence are popularly consumed by betel quid (BQ) chewers in Taiwan and many other South and Southeast Asian countries. However, little is known about the biochemical properties of inflorescence Piper betle (IPB) toward reactive oxygen species (ROS) and platelet functions. In the present work, aqueous IPB extract was shown to be a scavenger of H(2)O(2), superoxide radical, and hydroxyl radical with a 50% inhibitory concentration (IC(50)) of about 80, 28, and 73 microg/mL, respectively. IPB extract also prevented the hydroxyl radical induced PUC18 plasmid DNA breaks at concentrations higher than 40 microg/mL. Since ROS are crucial for platelet aggregation, we further found that IPB extract also inhibited the arachidonic acid (AA) induced and collagen-induced platelet aggregation, with an IC(50) of 207 and 335 microg/mL, respectively. IPB extract also inhibited the AA-, collagen- (>100 microg/mL of IPB), and thrombin (>250 microg/mL of IPB)-induced thromboxane B(2) (TXB(2)) production by more than 90%. However, IPB extract showed little effect on thrombin-induced aggregation. These results indicated that aqueous components of IPB are potential ROS scavengers and may prevent the platelet aggregation possibly via scavenging ROS or inhibition of TXB(2) production.

This double-blind randomized control study was conducted to evaluate whether a nonsteroidal anti-inflammatory drug (NSAID) could act as an effective pain control method during first trimester suction abortion, and whether co-treatment of NSAID with misoprostol will decrease the efficacy of the cervical ripening effect of misoprostol. Subjects were randomized to receive misoprostol alone or misoprostol together with diclofenac sodium. Both groups of subjects suffered from similar incidence of preoperative side effects. Co-treatment of NSAID with misoprostol did not attenuate the cervical ripening efficacy of misoprostol. There was no significant pain reduction in the group treated with NSAID, except that a marginal benefit was found in the subgroup of multiparous women. About two thirds of the subjects in both treatment groups found that this was a satisfactory pain relief method during the procedure.

Formocresol has long been used for pulpotomy of primary teeth and as an intracanal medicament. Little is known, however, about the pharmacological effect of tricresols. This study showed that three cresol isomers, o-cresol, m-cresol and p-cresol, are H2O2 scavengers with a 50% inhibitory concentration (IC50) of 502, 6.7 and 10.16 microM, respectively. o-, m- and p-cresol were also shown to be effective scavengers of superoxide radicals generated by xanthine/xanthine oxidase with an IC50 of 282, 153 and > 4000 microM, respectively, as analyzed by luminometer. o-, m- and p-cresol showed protective effects on the DNA breaks generated by H2O2/FeCl2 and FeCl3/ascorbate/H2O2 systems at concentrations ranging from 70 microM to 1.43 mM, o-, m- and p-cresol also showed differential protective effects against DNA breaks induced by 0.17% NaOCl with 100% inhibitory concentration (IC100) of about 10, 1 and 10 mM, respectively. In addition, reaction with 3% H2O2 and 0.17% NaOCl completely prevented NaOCl-induced DNA breaks. The results indicate that the three cresol isomers are effective ROS scavengers and may prevent ROS induced damage when used as pulpotomy agents or as intracanal medicaments. Owing to the difference in the position of the functional hydroxyl group in the three cresol isomers, m-cresol is the most effective ROS scavenger. Concomitant use of H2O2 for root canal irrigation may diminish both the tissue dissolving capacity of NaOCl and NaOCl-induced DNA damage.