The aim of this study was to assess the influence of an admission eGFR<60 mL/min/1.73 m2) and the development of AKI: eGFR≥60 without AKI, eGFR<60 without AKI, eGFR≥60 with AKI and eGFR<60 with AKI.A prospective study of 828 AMI patients was performed. AKI was defined as a serum creatinine increase of ≥50% from the time of admission (RIFLE criteria) in the first 7 days of hospitalization. Patients were divided into subgroups according to their eGFR upon hospital admission  for 30-day mortality were 2.00 (95% CI 1.11–3.61) for eGFR<60 without AKI, 4.76 (95% CI 2.45–9.26) for eGFR≥60 with AKI and 6.27 (95% CI 3.20–12.29) for eGFR<60 with AKI. Only an admission eGFR of <60 with AKI was significantly associated with a 30-day to 1-year mortality hazard .AKI development was associated with an increased early mortality hazard in AMI patients with either preserved or impaired admission eGFR. Only the association of impaired admission eGFR and AKI was associated with an increased hazard for late mortality among these patients. Recent North American and European epidemiological studies have shown that the incidence of acute kidney injury (AKI) is increasing at an alarming rate Ischemic heart disease is the leading cause of death among adults in high-income countries and accounts for a substantial fraction of the total disease burden globally. AKI is an important and common complication after acute myocardial infarction (AMI), affecting from 10 to 55% of the patients (this latter number referring to patients suffering from cardiogenic shock) The effect of pre-existing renal dysfunction on AKI mortality remains controversial and conflicting results have been published The purpose of this study was to evaluate the association of a decreased admission eGFR with the incidence and early and late mortality of patients developing AKI, as defined by the RIFLE criteria, in the acute phase of a myocardial infarction.2 (interquartile range: 47.5–81.3 mL/min/1.73 m2).We evaluated 828 patients with a median age of 65 years (interquartile range: 54 to 74), 65.5% of whom were male. Our study population was composed of 7.7% black patients and 92.3% non-black patients. Overall, at the time of the study, 69% of the patients had a history of hypertension, 36.7% smoked, 25.7% were diabetic, 22.6% were dyslipidemic, 41.7% had previously used angiotensin-converting enzyme inhibitors (ACEIs)/angiotensin II receptor blockers (ARBs), 8.8% had prior percutaneous coronary intervention (PCI), 15.5% had been affected by prior coronary artery disease (CAD) with greater than 50% stenosis, 16.3% had suffered from prior infarction and 8.5% of the patients had previously undergone coronary artery bypass graft (CABG) surgery. The median SCr of patients upon hospital admission was 1.2 mg/dl (interquartile range: 1.0–1.5 mg/dL), and the admission median eGFR was 63.1 mL/min/1.73 mThe systolic left ventricular function (LVF) was measured in 709 patients and was classified as normal in 29.7%, mildly dysfunctional in 28.6%, moderately dysfunctional in 22.1% and severely dysfunctional in 19.6% of them. ST elevation myocardial infarction (STEMI) was diagnosed in 50.2% of the patients, 20.9% of whom presented with a Killip class >I and 54.5% of whom presented with an anterior wall infarction. The overall in-hospital mortality was 13.6%.AKI occurred in 121 (14.6%) of the patients, with 76 patients of these patients (9.2%) in the Risk category, 37 patients (4.5%) in the Injury category and 8 patients (1%) in the Failure category. The 30-day mortality rate was 38.8% , and the 30-day to 1-year mortality rate was 25.4% .2 (interquartile range: 36.7–52.8 mL/min/1.73 m2). Patients with non-impaired eGFR had a median admission SCr of 1.0 mg/dL (interquartile range: 0.8–1.2 mg/dl) and a median admission eGFR of 79.2 mL/min/1.73 m2 (interquartile range: 68.5–96.1 mL/min/1.73 m2).Patients with an impaired eGFR had a median admission SCr of 1.5 mg/dl (interquartile range: 1.3–1.8 mg/dL) and a median admission eGFR of 46.1 mL/min/1.73 m2. When compared to the group of patients with an admission eGFR≥60 mL/min/1.73 m2, this group was composed of a smaller proportion of men; was significantly older; and had a significantly increased prevalence of hypertension, diabetes, prior infarction, prior use of ACEIs/ARBs and previously documented CAD, PCI or CABG. A significantly higher number of patients with an admission eGFR≥60 mL/min/1.73 m2 had an admission heart rate (HR) >100 beats/min and a systolic blood pressure (SBP) <100 mmHg. These patients were also more likely to have non-ST elevation myocardial infarction (NSTEMI). When presenting with STEMI, these patients exhibited a higher frequency of Killip class >I  3 flow rate or in the favorable reperfusion criteria after reperfusion treatment , were more likely to have a HR>100 beats/min (p = 0.013) and were more likely to have a history of diabetes (p = 0.009). Among patients with admission eGFRs<60 mL/min/1.73 m2, there were no significant differences between subgroups with and without AKI  for death of 2.00 (95% CI 1.11–3.61). Patients with a non-impaired admission eGFR and AKI had a death AHR of 4.76 (95% CI 2.45–9.26). The patients with an impaired admission eGFR and AKI had the worst outcome with a mortality AHR of 6.27 (95% CI 3.20–12.29). The reference group was composed of patients with an admission eGFR≥60 mL/min/1.73 m2 without AKI  .Previous studies have demonstrated that either an impaired renal function at admission or the subsequent development of AKI negatively affects the outcome of patients suffering from an AMI In this study, the use of Cox multivariate analysis clearly revealed that the development of AKI was the predominant factor associated with increased 30-day mortality, while an impaired admission eGFR was strikingly associated with an increased long-term mortality in patients with AMI-associated AKI. In fact, during the period from 30 days to one year after hospital admission, only the patients with an impaired admission eGFR and AKI had significantly lower survival rates. These results suggest that early mortality was largely related to the effects of AKI, while long-term outcomes were influenced by AKI development in addition to previously impaired renal function.2 were less likely to receive pharmacological therapy or to undergo interventional procedures. Other authors have also observed that AMI patients with an impaired admission eGFR were offered less AMI treatment The factors leading to the remarkably higher mortality among patients with an impaired admission eGFR and AMI-associated AKI are not evident. One possible explanation for these poorer outcomes may be the combination of a powerful AKI-induced surge of inflammation along with the high prevalence of cardiovascular risk factors, such as older age, diabetes, hypertension and a pro-inflammatory milieu already present in the group of patients with impaired admission eGFR A limited number of studies have assessed the role of admission eGFR on the outcome of AMI-associated AKI. A study on STEMI patients Overall, studies regarding the prognostic effect of pre-existing renal dysfunction on mortality after AKI have reported conflicting results. Some reported a decrease in early mortality in patients with overlapping AKI and CKD compared to the controls in unselected cohorts of critically ill patients In this study, there was no difference in the incidence of AKI between patients with an admission eGFR<60 mL/min or an eGFR≥60 mL/min. Similarly, Lombardi et al. Although not assessed in the present study, proteinuria, a marker of renal injury, has been recently identified as an important risk factor for the development of AKI in various situations, such as following cardiac surgery This was a single-center observational prospective cohort study.Systolic left ventricular function as assessed by echocardiography was not always determined by the same observer.The present study cannot be used to determine the effect of CKD on the incidence and mortality of AKI because the definition of CKD AKI incidence was probable underestimated. The reference SCr used for AKI diagnosis was the one obtained at the hospital admission. It is likely that some patients were hospitalized already with AKI, but because there was no SCr increase from the reference SCr during the hospital stay, they were misdiagnosed as non-AKI.Finally, SCr measurements were performed on a clinical need basis after discharge from the intensive care unit, which could also lead to an underestimation of the incidence of AKI.AKI development was negatively associated with the early outcomes of AMI in patients with either non-impaired or impaired admission eGFR. On the other hand, the presence of a low admission eGFR in patients with acute myocardial infarction-associated AKI was clearly associated with a significantly poorer long-term prognosis. Additionally, this study confirmed, using RIFLE criteria that AKI development in AMI patients is frequent and is associated with a high mortality rate.These results strongly suggest that patients with an impaired admission eGFR should have their renal function monitored closely after an acute myocardial infarction. Clinical measures for the prevention and early diagnosis of AKI must be taken to potentially increase the survival rates of these patients. Moreover, patients with an impaired admission eGFR who develop AKI must have an extremely careful and detailed long-term follow-up to try to avoid the poor long-term outcomes described in this study.The study protocol was approved by the Institutional Ethics Committee on September 21, 2009 , who agreed that informed consent was unnecessary due to the purely observational and non-interventional nature of this study (“Resolução CNS 196/96”).A total of 1,012 consecutive patients (October 2004–December 2008) with a STEMI or with a NSTEMI 2 or chronic dialysis (21 patients); death within 48 hours of admission (49 patients); lack of at least two SCr measurements in the first seven days of hospitalization (94 patients); obstructive AKI (3 patients); and a hospital stay <48 hours (17 patients).Among the 1,012 patients, 184 were not included in the study because they met the following exclusion criteria: admission SCr≥6.0 mg/dL, admission eGFR<15.0 mL/min/1.73 mOnly the first hospital admission was considered if a patient had more than one hospitalization for AMI during the study period.Systolic left ventricular function was classified as either normal function or mild, moderate or severe dysfunction and was assessed by echocardiography  in 85.6% of the patients The SCr level was obtained at the time of hospital admission and daily during the intensive care unit (ICU) stay. After ICU discharge, the SCr was measured as needed. The SCr was assessed using the Jaffé colorimetric method .2): GFR = 186.3×SCr −1.154 * age −0.203 * 1.212 (if patient is black) * 0.742 . It should be noted that this equation was not specifically validated for this study population.The admission eGFR was assessed by the Modification of Diet in Renal Disease (MDRD) formula Patients were divided into 4 subgroups:admission eGFR≥60 without AKIadmission eGFR<60 without AKIadmission eGFR≥60 with AKIadmission eGFR<60 with AKIPatients were diagnosed with AKI if they had a SCr increase of ≥1.5-fold over the admission value within the first seven days of hospitalization . The RIFLE criteria definition of AKI using urinary output (<0.5 mL/kg for 6 h for Risk) was not used in this study The primary endpoints were death from any cause within 30 days and 30-day to 1-year mortality for those patients who survived after 30 days. Patient follow-up after discharge was conducted either by an electronic hospital system records review or by mail or telephone contact of patients.2) and on whether they developed AKI during their hospitalization. The demographics and clinical characteristics  were compared by the Student's t-test, Mann-Whitney test or Kruskal-Wallis test followed by Dunn's post-test for continuous variables. The categorical variables (presented as numbers and percentages) were compared using either χ2 statistics or Fisher's exact test. Bonferroni corrections were used for post-test multiple comparisons.Patients were categorized based on their admission eGFR  and on the 30-day to 1-year mortality rates for those patients (n = 699) who survived past day 30. Patients who had been part of the follow-up period for less than one year were not included (n = 107) in the descriptive and univariate analyses of the mortality rates at that time point.We performed multivariate Cox proportional hazards regression analyses to evaluate the two mortality periods. The association between mortality and admission eGFR and AKI development in the four subgroups in which patients were divided was evaluated, controlling for clinically important variables and for variables with a p-value<0.15 in the univariate analyses of mortality in each period. The proportional hazards test and the plotted cumulative survival estimate after the ln (-ln) transformation suggested that the hazards of these variables were proportional for the period analyzed.In the first period of multivariate Cox proportional hazards regression analysis, the follow-up was censored at day-30 and was adjusted for age, creatine phosphokinase MB (CPK-MB) and admission glycemia (categorized by quartiles with the first as a reference), gender , history of prior CABG, STEMI, history of diabetes, history of hypertension, admission Killip class >I, SBP<100 mmHg, admission HR >100 beat/min, clopidogrel use during the hospitalization, use of diuretics, coronary angiography during the hospitalization, reinfarction, severe systolic left ventricular dysfunction (LVD) and any PCI performed during the hospitalization.The multivariate Cox proportional hazards regression analysis of 30-day to 1-year mortality rates included all patients who survived for 30 days after AMI with censoring at 365 days. The controlling variables were the same used for the 30-day analysis with the exception of CPK-MB, history of prior CABG, SBP<100 mmHg and reinfarction. The prior use of ACEIs or ARBs was also controlled in this analysis.Differences were considered statistically significant by a two-tailed p-value<0.05 and a confidence interval (CI) of 95%.Analyses were performed with SPSS statistical software .


Williams–Beuren syndrome  is a multisystemic neurodevelopmental disorder caused by a hemizygous deletion of 1.55Mb on chromosome 7q11.23 spanning 28 genes. Here we report a patient showing mild WBS physical phenotype, who carries a longer 21Mb atypical deletion.Genomic DNA from the proband was extracted from peripheral blood leukocyte. Karyotype analysis was performed on metaphase cells. Array-based comparative genomic hybridization of DNA from the patient’s peripheral blood lymphocytes was performed.The proband, a female neonate, is the first child of healthy nonconsanguineous Chinese parents. She was born by uterine-incision delivery with intrauterine distress after 41 weeks of gestation. Her birth weight was 2.4 kg. She showed an distinctive facies including broad brow, periorbital fullness, epicanthal folds, short nose, long philtrum, small jaw and prominent earlobes. The cardiology ultrasound examination showed open foramen ovale without elastin arteriopathy such as supravalvular aortic stenosis, pulmonic stenosis. Her abdominal ultrasound examination showed right Duplicated kidneys. Her Karyotyping was 46, XX, del(7)(q11.1q11.23). We then performed array CGH for this patient and confirmed the deletion region of 21Mb from 7q11.2 to 7q21.11.A deletion of the 7q11.23 chromosomal region is found in approximately 90-95% of the clinically typical WBS patients but in a lower percentage of atypical cases. The commonly deleted chromosomal region has a size of approximately 1.5 Mb. However, smaller or longer deletions have also been described in atypical WBS patients. Array CGH analysis can be performed to make sure the location and size of microdeletion and confirmed the diagnosis of patients with mild or atypical physical phenotypes.


We conducted a four-city time-series study to investigate acute effects of O3 and NO2 in the Pearl River Delta (PRD) of southern China, using data from 2006 through 2008.Background and objectives: Epidemiologic studies have attributed adverse health effects to air pollution; however, controversy remains regarding the relationship between ambient oxidants  and 1.95%  increases in total mortality, respectively, with stronger estimated effects for cardiovascular and respiratory mortality. After adjusting for PM10, estimated effects of O3 on total and cardiovascular mortality were stronger for exposure during high-exposure months (September through November), whereas respiratory mortality was associated with O3 exposure during nonpeak exposure months only.Results: We found consistent positive associations between ambient oxidants and daily mortality across the PRD cities. Overall, 10-μg/m3 and NO2 in the PRD and strengthen the rationale for further limiting the ambient pollution levels in the area.Conclusions: Our findings suggest significant acute mortality effects of O Many epidemiologic studies suggested both short- and long-term effects of exposure to O3 , using a time-series approach.Air monitoring data collected through the PRD monitoring network have made it possible to assess the health effects of ambient air pollution on a regional scale. In this multicity study, we estimated associations between ambient OStudy site description. Our study cities included Guangzhou, Foshan, Zhongshan, and Zhuhai in the PRD  deaths  and deaths attributed to cardiovascular diseases (codes I00–I99) and respiratory diseases (codes J00–J98) at all ages. The data also included subcategories of cardiovascular and respiratory mortality: coronary (codes I00–I09 and I20–I52), stroke (codes I60–I69), and chronic obstructive pulmonary disease .Environmental data. Air pollution data were measured at five monitoring stations in the area: two in Guangzhou , and one station each in Foshan, Zhongshan, and Zhuhai , carbon monoxide (CO), and PM10 (PM with aerodynamic diameter ≤ 10 μm) from each station. NO2, SO2, O3, and CO were measured using chemiluminescence, fluorescence, ultraviolet, and infrared instruments, respectively; PM10 was measured by tapered element oscillating microbalance. We calculated 24-hr average concentrations for PM10, NO2, SO2, and CO for days that had measurements for at least 18 of 24 hr. We calculated 8-hr (from 1000 hours to 1800 hours) average concentrations for O3 for days with at least six of eight hourly measurements available. If monitor data for an individual pollutant were insufficient to calculate a daily average, all measurements from that day were excluded for that pollutant and monitor. Data from the two monitoring stations were averaged to derive concentrations for Guangzhou. Daily temperature and RH data for each city were obtained from the Chinese Academy of Meteorological Sciences. Missing data were not imputed.We obtained hourly concentrations of NOStatistical analysis. Because daily mortality counts typically follow a Poisson distribution, we used Poisson regression models to evaluate the association between mortality and air pollution controlling for temperature, RH, seasonal patterns, and long-term trends using natural spline smoothers (moothers . Degreesmoothers  and genemoothers . Becausemoothers  suggest moothers . Residua3 increase in the average concentration of each pollutant during the previous 1–2 days. Single-day temperature and RH (lag 0 or lag 1 day) were used in our analyses, and the reported results were from models with lag 1 day covariates.Associations between mortality and average air pollutant concentrations on individual days (lag 0 to lag 6) and 2-day periods (lag 0–1 days and lag 1–2 days) were first examined in single-pollutant models. Preliminary analyses indicated that the largest pollutant effects were usually observed at lag 1–2 days (data not shown). Therefore, we report the excess risk (ER) of mortality and its 95% confidence interval (CI) associated with a 10-μg/mQ-statistics to test the homogeneity of effect estimates among the study cities (α = 0.05) (3 and from 0.43 to 0.83 for NO2), we also generated combined effect estimates by summing the mortality data across the four cities and averaging the environmental data from the five monitoring stations.After establishing the final model that controlled for time trend, temperature, RH, year, DOW, public holiday, and influenza epidemics, we calculated city-specific estimates by fitting Poisson regression models for each city separately. We calculated  = 0.05) . Because = 0.05) , and are3, NO2, and PM10 adjusted for confounding by other pollutants. Two-pollutant models were restricted to pollutants with Pearson correlation coefficients < 0.6 to avoid multicollinearity. Furthermore, we stratified O3 exposure as exposure during peak (September through November) and nonpeak (December through August) exposure periods identified previously for the area . Statistical significance was defined as p < 0.05.Analyses were performed using R  with the mgcv package , whereas NO2 was highly correlated with PM10, SO2, and CO . Temperature was positively correlated with O3 only, whereas RH was negatively correlated with all the pollutants  ranged from 0.22% to 0.64% for O3, 1.22% to 1.87% for NO2, and 0.37% to 0.74% for PM10 ]. We also observed significant increased risk in cardiorespiratory mortality per unit increase in air pollution: a 10-μg/m3 increase in O3 exposure was associated with 0.79% , 1.17% , and 1.16%  increases in mortality from coronary, stroke, and COPD diseases, respectively  and 66.9 μg/m3 in the nonpeak period. O3 exposure was significantly associated with total mortality and cardiovascular mortality in both periods . Altering the degrees of freedom of time and meteorological smoothers and excluding days with extremely high or low O3 concentrations did not alter total mortality effect estimates by > 20% . However, estimates for total mortality were somewhat sensitive to adjustment for temperature over different lag periods, resulting in ERs for a 10-μg/m3 increase in O3 (1- to 2-day lag) of 0.70%  and 0.63%  when adjusted for temperature with a lag of 2–3 days or 4–6 days, respectively, compared with 0.81%  when adjusted for temperature with a 1-day lag.Our sensitivity analyses indicated that covariates did not introduce collinearity, and all were significant predictors , and effect estimates for NO2 were robust to adjustment for O3. Effect estimates for O3 were larger for exposure during the peak period for total and cardiovascular mortality after adjustment for PM10, whereas for respiratory mortality, the association appeared to be limited to exposure during the nonpeak period.We estimated significant acute mortality effects associated with exposure to ambient oxidants in the PRD. In general, associations between O3 in the PRD cities  than those observed in North American cities (14–38 μg/m3)  increase in the previous week’s O3 was associated with a 0.52%  increase in daily mortality and a 0.64%  increase in cardiovascular and respiratory mortality  project . Considering the limited number of estimates available for meta-analysis, future studies should be conducted in more Asian cities in order to address the inconsistency.We observed much higher concentrations of O8 μg/m3) . Our anaortality . A meta-ortality . However project  and stud project . Wong et2 and mortality in the PRD, with estimated increases of 1.95%  in total mortality, 2.12%  in cardiovascular mortality, and 3.48%  in respiratory mortality, per 10-μg/m3 increase in lag 1–2 day NO2 concentrations. However, the magnitude of NO2 effect estimates in our study was much greater than comparable estimates reported for Western populations  .Consistent with previous studies , our anarations) . Our est3 and NO2 are major air pollutants in the PRD. Estimated effects of O3 were moderately reduced but still significant after adjustment for PM10, SO2, and CO in two-pollutant models. This is probably because O3, as a secondary pollutant, has formation paths in the environment different from those of PM10, SO2, and CO and thus does not typically covary with these pollutants. With weak correlations observed between O3 and other pollutants, the mortality effect of O3 exposure is at least partially independent of other pollutants, which is consistent with findings reported previously  generated by the same sources, such as vehicle emissions, or may reflect the combined effects of pollutants related to traffic and atmospheric photochemical pollution in the PRD. The significant associations estimated for O3 and NO2 suggest that ambient oxidants may play important roles in initiating air pollution-related mortality effects in the PRD.Ambient oxidants of Oeviously . Unlike 3 estimated for exposures during the peak and nonpeak exposure periods. The PRD lies on the border between subtropical and tropical zones, and the highest O3 concentrations often occur in autumn (September through November), in contrast with other geographic areas studied, where peak O3 period occurs in summer (June through August),  when the O3 level is higher , . Correlae summer . In the s higher . Althoug3 levels , 2001. Pr levels .2.5 and component data were available for the study period. Compared with PM10, PM2.5 is a better index of combustion source air pollution and has a larger proportion of secondary particles. 3 effects. Therefore, adjusting for PM10, instead of PM2.5 or its components, may overestimate the effect of O3.Limitations should be noted in interpreting the results of our study. First, exposure data were obtained from only one or two air pollution monitoring stations in each city. Air pollution exposure may be spatially autocorrelated across our study cities, which may violate the assumption of independent exposures among cities for the merged regression analyses. The limited number of air pollution monitoring stations and exposure data to assess spatial structure of air pollution between cities may yield biased variance of parameter estimates and inefficient significance tests in our study . Second,3 and NO2 exposures in the PRD. The evidence of differential effects of O3 on mortality from different diseases supports the need for further investigation of the pathophysiological mechanisms of O3-associated cardiovascular and respiratory effects. Our findings strengthen the rationale for further limiting ambient oxidant pollution in the PRD.We estimated significant increases in mortality associated with O(94 KB) PDFClick here for additional data file.


Iodine deficiency disorders (IDD) may affect the child growth and development. Speech development disorders in IDD area caused by brain damage. The incidence of speech developmental disorders is still high. Stimulation to children under 3 years old may increase speech capability due to brain plasticity. The aims of this study is to determine the effect of stimulation on speech development of 1-3 years old children in IDD and Non IDD areas.Quasi experimental study used one group pretest posttest design with consecutive sampling was done on 1- 3 years old children who fulfilled inclusion criteria in Kepil (Non IDD area) and Kertek District (IDD area), Wonosobo regency from April to September 2011. Stimulation interventions in accordance with the guidelines for implementation of stimulation, early detection and intervention for growth and child development in Primary Health Care 2006. Standard score equivalent of global language from Early Language Milestones Scale 2 was measured before and after stimulation in IDD and Non IDD areas. Statistical analysis used paired and independent t test.Eighty children consisting of 57.5% boys and 43.5% girls were enrolled in this study. Mean of speech development score in IDD 8,4  while in Non IDD 2,93  with p value 0.004. Standard score equivalent of speech development before and after stimulation in Non IDD area increase from 89.8 to 92.7 (SD 8.3) (p=0.032) and 85.7 to 94 (SD 7.94) (p=0.001) in IDD area. These result suggest that stimulation intervention has an effect on increasing speech development of 1-3 years old children in IDD and Non IDD area.


Stathmin is closely correlated with the progression and prognosis of a number of types of human cancer. The present study analyzed the associations between genetic variations in the stathmin gene and clinical outcomes of ovarian cancer. A total of 178 patients with epithelial ovarian cancer were treated with cytoreductive surgery followed by platinum-based chemotherapy. DNA was extracted from fresh tumor samples obtained during surgery. A total of 32 DNA samples were selected randomly for resequencing of the stathmin gene. Tag single nucleotide polymorphisms (SNPs) were identified based on the haplotype model as analyzed by PolyPhred software. Direct sequencing was employed in the genotyping of stathmin in 178 cases. A total of 10 nucleotide variations in stathmin were identified, of which 3 high-frequency variations were known SNPs from databases and 7 were new variations with low frequencies. The tag SNPs rs159531 and rs11376635 were selected from the linkage disequilibrium block of the gene to genotype stathmin in 178 cases. The distribution of the rs159531 genotype in ovarian cancer was 52.8% C/C, 35.4% C/T and 11.2% T/T. The distribution of the rs11376635 genotype in ovarian cancer was 32.0% G/G, 48.3% G/-, 18.5% -/-. The main haplotypes calculated by phase2.0 software were 55.6% CG, 27.8% T-, 15.4% C- and 1.2% TG. However, no associations between the stathmin genotype or haplotype and the outcomes in patients with ovarian cancer were observed. The stathmin genotype and haplotype were not associated with the phenotype of patients with ovarian cancer. Epithelial ovarian cancer is the leading cause of mortality from gynecological cancer worldwide and the 5-year survival rate remains <40% despite good initial responses to standard post-operative adjuvant chemotherapy with combined paclitaxel and cisplatin –3. The mStathmin is encoded by the human STMN1 gene located at chromosome 1p36.1. The encoded protein prevents the assembly and promotes the disassembly of microtubules and is involved in the regulation of the microtubule filament system by destabilizing microtubules . NumerouIn the present study, the stathmin gene was resequenced by genotyping tag single nucleotide polymorphisms (SNPs) based on haplotype model analysis, then the associations of the stathmin genotype with treatment responses and disease progression were analyzed.A clinical study of ovarian cancer was conducted at the Gynecologic Oncology Unit at University of Turin  between October 1991 and February 2000. An ethics review committee at the university approved the study and all participants provided informed consent. From the study, 178 patients who had primary epithelial ovarian cancer and received post-operative platinum-based chemotherapy were identified. The median age of the patients at surgery was 57.4 years, with a range of 26 to 82 years. Of the 178 patients, 33 (18.5%) were diagnosed with stage I disease, 12 (6.7%) were stage II, 120 (67.4%) were stage III and 13 (7.3%) were stage IV. The disease staging was classified according to the criteria of the International Federation of Gynecologists and Obstetricians (FIGO)  includedFollowing surgery, all the patients were treated with platinum-based chemotherapy; among them, 75 received platinum with paclitaxel (42.1%), while 103 received platinum without paclitaxel (57.9%). At 1 month after the chemotherapy, the treatment response was assessed based on the WHO criteria . A complhttp://primer3.wi.mit.edu/). The primer sequences for PCR are shown in μl mixture was prepared for each reaction and included 1X HotStarTaq buffer, 2.0 mM Mg2+, 0.2 mM dNTP, 0.2 μM of each primer, 1 unit HotStarTaq polymerase (Qiagen) and 1 μl template DNA (5–10 ng/μl). The cycling program was 95°C for 15 min; 11 cycles of 94°C for 15 sec, 62-0.5°C per cycle for 40 sec, 72°C for 1 min; 24 cycles of 94°C for 15 sec, 56°C for 30 sec, 72°C for 1 min; and 72°C for 2 min. The PCR products were purified with shrimp alkaline phosphatase enzyme  and exonuclease I . Cycle sequencing was performed with the BigDye Terminator Cycle Sequencing Ready Reaction kit  on an ABI 3130 Genetic Analyzer (Applied Biosystems). The primer sequences for sequencing are shown in http://droog.mbt.washington.edu/poly_get.html). Pairwise linkage disequilibrium (LD) between the sequence variations was analyzed by two parameters, |D’| and r2. The haplotypes from the sequence variations were reconstructed with the PHASE 2.1 program (http://www.stat.washington.edu/stephens/phase/download.html).Genomic DNA was extracted from tumor samples following manual homogenization using the QIAamp1 DNA Mini kit . A total of 32 DNA samples were randomly selected for resequencing from 1 kbp upstream to 1 kbp downstream of the stathmin gene. The entire coding exon and the flanking intronic sequences of the stathmin gene, covering 8.7 kbp, were amplified by PCR using the 16 pairs of primers designed by the online Primer3 software  and corresponding 95% confidence intervals (CIs) were calculated using the logistic regression model. By combining the data of stathmin expression from our previous study , the assAs shown in 2 showed these 10 variation loci were in high LD and the entire gene was in an LD block. Based on parameters including r2>0.8 and allelic frequency >10%, 2 tag SNPs were identified, rs159531 (T/C) and rs11376635 (-/G). According to the 2 tag SNPs, the main haplotypes were calculated as CG, T- and C-.The analysis of pairwise LD and rUsing direct sequencing in 178 patients with ovarian cancer, 2 tag SNPs with high allelic frequency, rs159531 (G/T) and rs11376635 (T/C), were identified for genotyping. The genotypes of rs159531 were 52.8% C/C (94/178), 35.4% C/T (63/178), 11.2% T/T (20/178) and 0.6% missing (1/178). The genotypes of rs11376635 were 32% G/G (57/178), 48.3% G/- (86/178), 18.5% -/- (33/178) and 1.1% missing (2/178). The distribution of the main haplotypes were 79.8% CG (142/178), 9.6% T- (17/178), 9.0% C- (16/178) and 1.7% TG 3/178; .As shown in No significant differences in the survival of patients with ovarian cancer according to the genotypes of stathmin were observed.By combining the present data with our previous mRNA expression of stathmin data, no significant correlations between the genotype and phenotype of stathmin were observed, as shown in Stathmin, first identified as a cytosolic phosphoprotein in neuroendocrine cells , has beeIn order to observe the possible biological function of those variations, 2 tag SNPs with high allelic frequency (rs159531 and rs11376635) were selected for genotyping and the associations between the genetic variants of stathmin and clinical characteristics of 178 patients with ovarian cancer were analyzed. The genotypic frequencies of these 2 tag SNPs were similar to the data from the dbSNP database. The majority of patients  had the CG haplotype. However, the genotype of the tag SNP or haplotype was not associated with the expression level of stathmin and no significant associations among genotype, clinical characteristics and outcomes in patients with ovarian cancer were observed.et al also analyzed rs182455 in 647 clinically well-characterized multiple sclerosis (MS) patients and 519 healthy controls, but no associations of the genotype of rs182455 SNPs with MS susceptibility or clinical disease course were observed (Stathmin is a microtubule-destabilizing protein that regulates microtubule dynamics by preventing tubulin polymerization and promoting microtubule disassembly during cell cycle progression ,30. Chanobserved .In conclusion, although no significant associations were observed between the genotype of stathmin and clinical characteristics or outcomes in patients with ovarian cancer in the present study, the results provided information concerning somatic or genetic variations of stathmin in patients with ovarian cancer.


Bacillus thuringiensis var. israelensis (Bti) is a suitable agent for controlling Chironomus kiiensis, a major pest polluting water. In this study, laboratory bioassays were used to study toxicity and affecting factors of Bti on C. kiiensis larvae. Tests were conducted using three commercial Bti formulations  of Bti. The toxicity of Bti formulations to third and fourth instar C. kiiensis larvae was in decreasing order of technical material, oil miscible suspension, and wettable powder, based on the 12 and 24 hour LC50 values. Increasing larval densities (from 10 to 30 per bioassay cup) increased the LC50 values for fourth instar C. kiiensis larvae. The LC50 values for fourth instar larvae reared in sand substrate were higher than those from soil substrate, and autoclaved substrates significantly increased the LC50 values. The technical material of Bti at 12 and 24 hours responded similarly to changes in temperature between 30° C and 15° C, but the LC50 values at a range of tested temperatures showed distinct differences in time points. Chironomus kiiensis Tokunaga has been reported to be the most prevalent chironomid species in Korea out of about 50 species recorded to date  has been used to control these insects. Lei et al.  is a species-rich family of flies, with almost 15,000 species described worldwide . As an i to date . In Chine vector . Since 2i et al.  investighttp://www.btrdc.com/).Oil miscible suspension , wettable power , and technical material  of Bti were obtained from Hubei Kangxin Agro-industry Co., Ltd. . Tests were conducted in 100-mL plastic cups containing 50 mL of test solution, which was comprised of dechlorinated water. Test vessels were not aerated and test solutions were not renewed for 24 hours. The test vessels were covered with clear plastic film to minimize evaporation. Tests were conducted at room temperature with a photoperiod of 16:8 L:D. The number of immobilized midges was recorded at 12 and24 hours using a dissecting microscope (× 10 magnification). Immobilization was defined as the cessation of all visible signs of movement or activity when viewed under the dissecting microscope. Each experiment was repeated using a different batch of midge larvae. An experiment was considered valid if mortality in the control did not exceed 10% at the end of the test and dissolved oxygen did not fall below 20% saturation. Bioassays evaluating the effects of larval age were conducted by measuring the toxicity of oil miscible suspension , wettable powder , and technical material  of Bti on third and fourth instar C. kiiensis larvae were susceptible to wettable powder, oil miscible suspension, and technical material of Bti. The 12 and 24 hour LC50S of the three Bti formulations showed similar changes for third and fourth instar C. kiiensis larvae and were in decreasing order of wettable powder, oil miscible suspension, and technicial material. The young third instar larvae were more susceptible than fourth instar larvae.The larvicidal activity of three Bti preparations is summarized in 50 of technical material on fourth instar larvae. The 12 and 24 hour LC50S of Bti with a density of 30 larvae/cup were 2.40 and 2.08 fold higher than at a density of 10 larvae/cup, respectively. The results showed that Bti activity in low densities was higher than in greater densities.C. kiiensis larvae. The 12 hour LC50S of Bti on fourth instar C. kiiensis larvae residing in autoclaved soil, sand, and autoclaved sand were increased by 10.43, 9.50 and 15.39 fold compared to 10 larvae per cup in soil, respectively. Moreover, the 24 hour LC50S of the fourth instar C. kiiensis larvae in autoclaved soil, sand, and autoclaved sand were also significantly higher than in soil substrate. However, the autoclaved substrates significantly influenced the toxicity of Bti on the C. kiiensis larvae. After 12 hours, the LC50S for larvae from autoclaved soil and autoclaved sand were 10.43 and 1.62 fold higher than those from soil and sand, respectively.50 values for technical material of Bti bioassayed against fourth instar C. kiiensis larvae  at a range of constant temperatures. The results showed that Bti at two time points responded similarly to changes in temperature between 30° C and 15° C. Between 30° C and 27° C, the Bti at 12 hours showed a steep (3.60 fold) decline in activity, whereas the corresponding decline in activity at 24 hours was only 1.02 fold. The 12 hour LC50 values at the tested temperature extremes for Bti were 3.11 mg/L (30° C) and 25.51 mg/L (15° C), while 24 hour LC50 values were 0.41 mg/L (30° C) and 2.25 mg/L (15° C). The LC50 values at a range of tested temperatures showed a distinct difference in time points, such as the 12 hour LC50 being 23.49 fold higher than the 24 hour LC50.To investigate the influence of temperature on the toxicity of Bti, five temperatures were selected to measure LCChironomus tepperi  ranging from 0.13 to 0.37 mg/L for third instar larvae, and from 0.32 to 1.57 mg/L for fourth instar larvae. These results were similar to mosquitoes . Our results showed a similar variation, in which the 12 and 24 hour LC50S of Bti at a density of 30 larvae/cup were 2.40 and 2.08 fold higher than at a density of 10 larvae/cup, respectively. Additionally, substrate type and temperature also affected Bti efficacy. Our data demonstrated that the LC50 values (fourth instar larvae) using sand substrate were higher than those in soil substrate. The characteristics of soil inhabited by chironomid larvae may therefore affect the Bti activity. Charbonneau et al. (50 values, thus suggesting that an increased toxicity in treated substrates might be due to the contribution of other microorganisms in original substrates. However, Stevens et al. (Since Bti was initially described and applied to control mosquito larvae , Bti-bas tepperi . Formula tepperi . The chisquitoes  and othesquitoes , with thsquitoes  also repu et al.  demonstrs et al.  found sos et al. . TherefoC. tepperi has been shown to display large variations in LC50 values at 15° C. In this study, the data with technical materials of Bti and C. kiiensis larvae showed similar changes at temperatures between 30° C and 15° C. The Bti toxins can produce toxicity in the insect midgut after ingestion (Temperature is also a major factor affecting Bti toxicity . The toxngestion , and it ngestion . To effe


P < 0.01) but no significant reduction in body fat percentage (P < 0.05). The findings suggest that weight reduction does not promise a reduction in body fat. Changes in neck circumference were only found in normal subjects. Hence, it can be said that overweight and obese subjects showed no changes in anthropometry status during Ramadan. No changes in body composition were reported in all three weight groups except for trunk body fat. In conclusion, normal subjects showed significant changes in various anthropometry parameters, but overweight and obese subjects showed no obvious difference.This study was done to observe the anthropometry and body composition changes before, during, and after the holy month of Ramadan. This study was carried out on 46 staff from one of the local universities, which comprised of 14 males and 32 females ranging in age from 25 to 40 years old. There were four sessions done to complete this study, namely, a week before Ramadan (T1), 1st week of Ramadan (T2), 3rd week of Ramadan (T3), and a month after Ramadan (T4). All subjects were assessed according to weight, body circumference, and body composition status. It was found that subjects with different weight status showed a significant reduction in weight ( Ramadan is the ninth month of the Islamic calendar where it is compulsory for all Muslims to abstain from eating, drinking, and smoking (just to name a few of the restrictions related to this study) from sunrise to sunset . In biocP > 0.05) in weight and BMI during the fasting period (26.86 kg/m2) compared to the period before the holy month of Ramadan (25.77 kg/m2).During the nonfasting months, Muslims usually consume approximately 3-4 meals in a day. Hence, frequency and quantity of food intake may be reduced during the holy month of Ramadan. During the holy month of Ramadan, dietary intake as well as the level of physical activity is drastically reduced as most Muslims consume at least two meals per day and restrict their physical activities. Commonly, Muslims have two heavy meals during Ramadan, one just before sunrise and the other immediately after sunset or also known as the breaking of the fast . El Ati Fedail et al.  also repThis study was done among 55 healthy Muslim adults aged between 25 and 40 years who were recruited for the first session of this study. However, nine subjects' dropped out from the study and only 46 subjects remained for the 2nd session until the 4th session of the study. The subjects were recruited from the UniSZA community who had responded to our invitation to take part in the study. This study was done among UniSZA staff from the Gong Badak campus between July and October 2012. A total of 46 subjects from UniSZA who had volunteered for this study had participated in this study session before, during, and after the holy month of Ramadan. The final sample size for this study was 46 subjects  with a mean age of 33.04 ± 4.57. The nominal inclusion criteria are ages between 25 and 40 years old, apparently healthy, metabolically normal (no metabolic problems), and a Muslim who practices fasting during the holy month of Ramadan. Exclusion criteria include presence of chronic illnesses, having metabolic syndrome, on drug treatment, or being pregnant. The study protocol was approved by the Medical Research Ethics Committee, Universiti Sultan Zainal Abidin.Anthropometry assessment for height, waist circumference (WC), neck circumference (NC), and mid-upper arm circumference (MUAC) was assessed using a portable stadiometer (SECA 217) and body circumference tape (seca203). While weight and body composition including body fat %, whole subcutaneous fat (WSF), whole skeletal muscle (WSM), trunk subcutaneous fat (TSF), trunk skeletal muscle (TSM), arm subcutaneous fat (ASF), arm skeletal muscle (ASM), leg subcutaneous fat (LSF), and leg skeletal muscle (LSM) were measured using the Karada Scan HBF-362 (OMRON). sahur  to ensure good hydration for accurate reading of body composition. All anthropometry and body composition assessments were done in four sessions, represented by T1, T2, T3, and T4. The 1st session of data collection (T1) was done a week before the holy month of Ramadan began. The second session of data collection (T2) was done a week after Ramadan (1st week of Ramadan) and the third session (T3) was done on 3rd week of Ramadan. While the last session of data collection (T4) was done a month after Ramadan . Data was collected a month after Ramadan because some subjects continued another 6 days of fasting after Ramadan had ended. Hence, to ensure that all subjects could attend the data collection sessions, the last data collection was done a month after Ramadan .Subjects were advised to drink adequate water during The data collected was analyzed by using Statistical Program Package for Social Sciences (SPSS) version 17.0. Descriptive statistical was performed to obtain mean and standard deviation. The level of significance was set at 0.05.Total of 46 subjects had participated conclusively in this study. Among the 46 subjects, 19 (41.3%) of them comprising 2 males and 17 females had a normal BMI, 18 (39.1%) of them  were overweight and another 9 (19.6%) of them  were obese. There were no subjects in the underweight category. All subject were Malay with a mean age of 34.3 ± 2.8 for males and 32.5 ± 5.1 for females. Mean body mass index (BMI) for males was 28.8 ± 3.4 and for females was 24.9 ± 3.5.Anthropometry status among subjects with different weight status during the holy month of Ramadan was compared as shown in P > 0.05). Waist circumference (WC) in normal subjects showed a significant decrease before the holy month of Ramadan (72.23 ± 6.39), during the 1st week of the holy month of Ramadan (71.38 ± 6.28), and after the holy month of Ramadan (71.29 ± 6.31) with P < 0.001. WC on overweight and obese subjects also showed a significant difference from T1 (P < 0.001) with 82.13 ± 6.74 (overweight) and 91.91 ± 6.37 (obese) and on T4 (P < 0.05) with 83.22 ± 10.96 (overweight) and 90.62 ± 8.67 (obese). On T2 and T3, WC on overweight and obese subjects showed no significant difference statistically (P > 0.005). However, the slight reduction in body fat among all weight groups was not statistically significant (Mid-upper arm circumference (MUAC) status among all groups  was significantly different from T1 until T4. In the normal weight group, MUAC was slightly increased from 27.15 ± 3.02 (T1) to 27.34 ± 2.86 (T2) and reduced to 26.26 ± 2.77 (T3) but increased slightly (27.02 ± 3.42) after the holy month of Ramadan (T4). In the overweight group, MUAC was reduced to 31.06 ± 1.99 (T2) compared to 31.83 ± 2.36 (T1) and increased to 31.10 ± 2.02 (T3) but reduced back to 30.99 ± 2.48 after the holy month of Ramadan (T4). MUAC in the obese weight group also increased a week into the holy month of Ramadan (T2) with 35.37 ± 2.59 compared to 34.47 ± 2.74 a week before the holy month of Ramadan (T1). However, MUAC in the obese weight group continuously reduced on T3 (34.58 ± 2.45) and T4 (34.39 ± 2.91).Neck circumference (NC), which has recently been used as a screening tool for obesity, showed no significant difference statistically on overweight and obese weight groups. However, NC showed significant difference before, during, and after the holy month of Ramadan for normal weight group, with 33.13 ± 3.29 (T1) and then reduced to 32.55 ± 2.77 (T2). NC in the normal weight group continued to decrease on T3 (32.12 ± 2.76) but slightly increased to 32.23 ± 3.00 (T4).Both, normal and overweight subjects showed a reduction in weight during the holy month of Ramadan and this increased slightly after the holy month of Ramadan but was still lower compared to the weight before the holy month of Ramadan. Obese subjects showed a continuous reduction of weight before the holy month of Ramadan and until a month after the holy month of Ramadan. A study done by Ramadan  among obThere is a lot of arguments on how weight reduction is interpreted during the holy month of Ramadan. Many studies have reported that calorie intake during the holy month of Ramadan increased compared to other months. Sugary food and drinks are consumed more frequently during the holy month of Ramadan . It is sThere was no difference in body fat (%) before, during, and after the holy month of Ramadan among all weight groups even though some weight reduction was indicated. An increase in body fat percentage is usually coupled with an increase in weight . Howeverr = 0.67) compared to BMI.Many studies have reported a reduction in waist circumference after the holy month of Ramadan , 19, 20.Body composition status is one important parameter that might change during fasting. Based on P < 0.05). TSM in obese subjects was lower compared to normal and overweight subjects. Based on the TSF in normal, overweight, and obese subjects showed a reduction a month after Ramadan  compared to a week before Ramadan . Reduction of TSF in normal and obese subjects was significant  was significantly higher compared to body fat percentage (P = 0.03); neck circumference (NC) also showed significantly (P = 0.006) greater reduction after Ramadan compared to body fat. This may suggest that reduction in NC is indicative of a reduction in weight.Generally, there were changes in body weight, body fat percentage, and neck circumference among subjects during the holy month of Ramadan. All these three parameters showed significant reductions after Ramadan compared to a week before Ramadan. However, there were no significant reductions in waist circumference and neck circumference. Reduction in weight  and mid-upper arm circumference (MUAC) showed a significant reduction after the holy month of Ramadan in all subjects in the different weight groups. However, overweight and obese subjects showed no significant changes in WC during the holy month of Ramadan; only normal subjects showed continuous changes in WC throughout Ramadan. Normal subjects also showed a continuous reduction in MUAC during and after the holy month of Ramadan. Overweight and obese subjects only showed a significant reduction on the first week of the holy month of Ramadan; no significant changes were seen in the third week of the holy month of Ramadan. NC in normal subjects continued to decrease throughout the holy month of Ramadan but increased slightly after Ramadan but was still less than the NC measured before the holy month of Ramadan. Even though there was a reduction of body fat after the holy month of Ramadan compared to before it, there was no change in body composition observed in this study. Normal and obese subjects showed a reduction in WSF, TSF, ASF, and LSF. Overweight subjects showed a reduction in TSF and ASF but no change in WSF and LSF. In terms of muscle percentage, normal subjects showed an increased in WSM, LSM, and TSM but no change in TSM. Overweight subjects only showed an increase in WSM but no changes in TSM, LSM, and ASM. Obese subjects showed no changes in ASM and LSM but showed an increase in WSM and TSM.In conclusion, only weight reduction, WC, and MUAC were indicated after the holy month of Ramadan in all weight groups, while reduction in NC was only seen in the normal weight group. There was no improvement seen in body composition status during Ramadan in this study except TSF, even though some studies have reported otherwise. Reduction in TSF was only seen in normal and obese groups. Hence, it can be said that fasting during the holy month of Ramadan brings no significant difference in body composition. However, fasting during the holy month of Ramadan may provide improvements to general health, body structure, and spiritual enhancement if the holy month of Ramadan is celebrated modestly with moderate food intake. If one appreciates the holy month of Ramadan, the quality of life may improve to the fullest. Since a high percentage of subjects that responded to our invitation were in the normal, overweight, and obese weight category, the effect on anthropometry and body composition during the holy month of Ramadan among underweight subjects could not be observed. Most of the staff of UNiSZA are already preoccupied with the dissemination of health information and are living a healthy lifestyle; hence, the probability of recruiting an underweight subject was severely reduced. More females were interested in participating in this study compared to males, which resulted in higher number of female subjects participating compared to males.Another limitation is the period allotted to carry out this study, since the holy month of Ramadan only lasts for approximately 30 days in a year. The subjects who responded in a week after Ramadan (second session) could still be accounted for and observed but subjects who came in for the third session (third week of Ramadan) but absent during the second session and vice versa need to be excluded from study.


Background: Combination therapy with antihypertensive agents utilises different mechanisms of action and may be responsible for a more effective decrease in blood pressure. Objective: To review the recently published trials on efficacy and safety of the combination therapy with olmesartan and amlodipine.  Results: The double-blind American COACH (Combination of Olmesartan Medoxomil and Amlopdine Besylate in Controlling High Blood Pressure) study (2008) showed in 1,940 patients that after eight weeks of treatment the BP goals were most frequently achieved in the ‘combination therapy group’, with 56.3% (54.1–58.5%) and 54.0% (51.8–56.2%) of patients reaching adequate blood pressure of <140/90 mmHg with olmesartan/amlodipine 20/10 and 40/10 respectively. Combination therapy was generally well tolerated. The most common side effect was oedema . The frequency of oedema was lower in the groups combining amlodipine 10 mg with olmesartan 10 mg , 20 mg  or 40 mg . In 2009 three double-blind controlled European studies including 500–1,000 patients each and performed independently of one another have confirmed the above study, and have demonstrated similar efficacy-safety effects from the combination of olmesartan medoxomil with amlodipine, particularly for patients not achieving adequate blood pressure control with olmesartan monotherapy. Conclusions: Combinations of olmesartan and amlodipine were significantly more effective at reducing blood pressure and realising guideline blood pressure goals in patients with mild to severe hypertension than monotherapy (with a placebo component). Combination therapy is well tolerated and is associated with a lower incidence of side effects, such as oedema, compared to monotherapy with high amlodipine dosages (10 mg).  According to the most recent data from the National Health and Nutrition Examination Survey, the prevalence of (age adjusted) hypertension in the American population, estimated from 14,653 individuals, is 29.3% (28.6–30.0%)  and is increasing . The. The3]. A meta-analysis of 61 prospective, observational studies of over one million patients in total showed that there was a 7% 6.95–7.05%) risk reduction in cardiovascular mortality for every 2 mmHg decrease in systolic blood pressure (SBP) and even a 10% 9.94–10.06%) reduction in the risk of death from cerebro-vascular accidents  3,4]. I,4. I3,4]–10.06% r–7.05% riOne of the most significant reasons for poor control is insufficient medical treatment to intensify antihypertensive therapy in patients unable to attain the blood pressure goals . Extra aVarious extensive studies have demonstrated that it is certainly possible to achieve blood pressure goals in patients with hypertension. However, the great majority will eventually require two or more antihypertensive agents to achieve this . Both thCombination therapy with antihypertensive agents utilises different mechanisms of action and may be responsible for a more effective decrease in blood pressure . One of The treatment of hypertension increasingly involves a combination of an ARB and a dihydropyridine calcium antagonist, and, in the past few years, fixed drug combinations have been used in a dose that is below the dose normally prescribed when each drug is used on its own . The com®) against monotherapy in patients with mild to severe hypertension [The purpose of the COACH study (Combination of Olmesartan Medoxomil and Amlodipine Besylate in Controlling High Blood Pressure) was to investigate the effectiveness and the safety profile of combinations of olmesartan and amlodipine (Sevikarrtension . The stu54.3% (52.1–56.5%) of the 1,940 randomised patients were male. The average age was 54 years and 19.8% were over 65 years of age. The average blood pressure at the start of the study was 164/102 mmHg, and 79.3% of patients had stage 2 hypertension. The combination therapy with olmesartan/amlodipine produced decreases in diastolic blood pressure (from –14.0 mmHg with olmesartan/amlodipine 20/5 mg to –19.0 mmHg with olmesartan/amlodipine 40/10 mg) and in systolic blood pressure  and 54.0% (51.8–56.2%) of patients reaching adequate blood pressure of <140/90 mmHg with olmesartan/amlodipine 20/10 and 40/10 respectively. Combination therapy was generally well tolerated. The most common side effect was oedema (olmesartan 20 mg 9.9% (8.6–11.3%), amlodipine 10 mg 36.8% (34.7–39.0%), placebo 12.3% (10.9–13.8%)). The frequency of oedema was lower in the groups combining amlodipine 10 mg with olmesartan 10 mg , 20 mg  or 40 mg .These findings of dose-dependent effectiveness with a combination of olmesartan and amlodipine are consistent with the results both from comparable studies of other fixed-dose combinations and from other studies specifically investigating a combination with a different ARB: valsartan with amlodipine .There are a number of fixed-dose antihypertensive combinations, such as ARBs combined with a diuretic agent (hydrochlorothiazide) and ACE inhibitors combined with a diuretic or a calcium antagonist. The treatment that underlies these combinations is consistent with the JNC 7 recommendation, which stipulates that combinations should comprise of antihypertensive constituents that work through different mechanisms of action in order to achieve maximum therapeutic effect . Both thFixed-dose combinations could be regarded as more patient friendly than taking two medicinal products separately. A recent meta-analysis demonstrated that compliance with fixed-dose antihypertensive combinations was significantly better than compliance with two separate medicinal products . actively investigates the development and the severity of oedema [When a higher dose of amlodipine was combined with increasing dosages of olmesartan, the lower incidence of oedema in the COACH study was consistent with other studies, where the use of a RAAS blocker combined with amlodipine reduced the probability of amlodipine-induced oedema ,15,16,17f oedema . Consequet al. reported 19% (16.3–21.6%) peripheral oedema while using amlodipine 5 or 10 mg in a study with a specific questionnaire actively asking after these symptoms [One study of valsartan and amlodipine, which followed a passive recording system , showed that 8.7% (2.1–26.5%) of patients developed oedema while using amlodipine monotherapy and 3.0% 0–17.2%) while using placebo .2% while. In contsymptoms . Another study (the CASTLE study), which followed a proactive monitoring system, even showed that 22.1% (16.8–27.0%) of patients who received amlodipine 5 or 10 mg had oedema . In the In the last year three double blind controlled European studies including 500–1,000 patients each and performed independently of one another have confirmed the above studies, and have demonstrated similar efficacy-safety effects from the combination of olmesartan medoxomil with amlodipine, particularly for patients not achieving adequate blood pressure control with olmesartan monotherapy ,22,23. WIn conclusion, combinations of olmesartan and amlodipine were significantly more effective at reducing blood pressure and realising guideline blood pressure goals in patients with mild to severe hypertension than monotherapy (with a placebo component). Combination therapy is well tolerated and is associated with a lower incidence of side effects, such as oedema, compared to monotherapy with high amlodipine dosages (10 mg).


Hand hygiene is the single most important measure for preventing healthcare associated infection. In Hong Kong the WHO hand hygiene program was launched in a territory wide healthcare system.In 2006, hand hygiene (HH) was piloted in 4 hospitals that included intervention and control wards. The promotion program included education, placement of Alcohol Hand Rub (AHR) at point of care and posters. In 2009, the program was extended to 74 out patient clinics, 14 Tradition Chinese Medicine clinics and 8 long-term Care Facilities.In 2008, “clean care is safer care” was implemented in 38 public Hospitals. A huge banner sized 9 x 15 meter was posted at the headquarter building. Same design was also put up across all hospitals. New designs were changed annually.Compliance rate of control wards was 22.16 % at baseline that increased to 25.27 %. For test wards it was 23.8% that significantly increased to 55.7 % and Chi-square test was <0.05.For the 74 out patient clinics, 14 Tradition Chinese Medicine clinics and 8 long-term Care Facilities, the overall compliance was 91%, 90% and 80% respectively.AHR was also measured as a surrogate marker of hand hygiene compliance. The amount used was found to have steadily increased from 3.2 L in 2007 to 12.9 L per 1000 patients bed days in 2010 and the correlation will be done when the data of 2011 is available.The WHO Hand Hygiene program can be successfully implemented in different kinds of healthcare by a concerted territory-wide effort.None declared.


Water flooding plays an important role in recovering oil from depleted petroleum reservoirs. Exactly how the microbial communities of production wells are affected by microorganisms introduced with injected water has previously not been adequately studied. Using denaturing gradient gel electrophoresis (DGGE) approach and 16S rRNA gene clone library analysis, the comparison of microbial communities is carried out between one injection water and two production waters collected from a working block of the water-flooded Gudao petroleum reservoir located in the Yellow River Delta. DGGE fingerprints showed that the similarities of the bacterial communities between the injection water and production waters were lower than between the two production waters. It was also observed that the archaeal composition among these three samples showed no significant difference. Analysis of the 16S rRNA gene clone libraries showed that the dominant groups within the injection water were Betaproteobacteria, Gammaproteobacteria and Methanomicrobia, while the dominant groups in the production waters were Gammaproteobacteria and Methanobacteria. Only 2 out of 54 bacterial operational taxonomic units (OTUs) and 5 out of 17 archaeal OTUs in the injection water were detected in the production waters, indicating that most of the microorganisms introduced by the injection water may not survive to be detected in the production waters. Additionally, there were 55.6% and 82.6% unique OTUs in the two production waters respectively, suggesting that each production well has its specific microbial composition, despite both wells being flooded with the same injection water. The results showed that most microorganisms in the injection water could not be detected in the production water, and each production well was composed of a unique microbial community.2. The viscosity of the crude oil was 400–2,000 mPas. Three samples were used in this study: one injection water (W), one production water (C) abstracted from oil-bearing stratum Ng44, and one production water (L) abstracted from oil-bearing strata Ng34 and Ng35. Physicochemical characteristics of the injection water and production waters are presented in The Gudao petroleum reservoir is located at Dongying in the Yellow River Delta of China, near the Bohai Sea. This oil field has been water flooded for over 30 years. The water content of production water in this reservoir is over 95%. The depths of the sampled petroleum horizons ranged from 1,173 m to 1,230 m, with a temperature of 69.5°C and a pressure of 12 MPa. The porosity of the reservoir was 33%, and air permeability was between 1.5–2.5 mDGGE analysis of a PCR-amplified V3 region of bacterial 16S rRNA genes showed that three dominant bands  were found in all three samples, and two dominant bands (d and e) were observed in C and L only. Several bands  were found in W but absent in C and L . StatistThree bacterial clone libraries  were constructed using total genomic DNA. From these libraries, 360 bacterial clones were randomly selected and sequenced . After dUsing the same analytical methods, three archaeal clone libraries  were constructed, from which 166 clones were randomly selected and sequenced . After dPseudomonas (24.3%), Thauera (5.2%), Marinobacterium (6.7%) and Pusillimonas (3.1%) had its abundance higher than 3.0%. Unlike BW, except for one OTU (B-OTU65), the remaining bacterial sequences in BC and BL belonged to Gammaproteobacteria (Pseudomonas (67.7%), Psychrobacter (29.3%) and Acinetobacter (1.5%). In BL, all of the sequences were assigned to three families, Moraxellaceae (97.1%), Pseudomonadaceae (1.5%) and Comamonadaceae (1.4%) (Psychrobacter (73.9%), Pseudomonas (1.4%), Acinetobacter (17.4%) and Acidovorax (1.4%), respectively.The compositional differences in bacterial communities were analyzed based on bacterial 16S rRNA gene sequences obtained from the three bacterial clone libraries . In BW, 0.5% sequences were considered phylogenetically undefined bacteria, which shared less than 75% identity with reference sequences in the database. The remaining sequences were clustered within five phyla: Proteobacteria (91.2%), Deferribacteres (3.5%), Bacteroidetes (2.2%), Thermotogae (1.6%) and Firmicutes (1.0%) . At the bacteria . In BC, e (1.4%) . Among tPseudomonas stutzeri (AF094748) (>98%), had an abundance of 1.6% in BW and 66.7% in BC but was not detected in BL. In previous literature, P. stutzeri has been shown to enhance crude oil biodegradation and increase rhamnolipid production Pseudomonas xanthomarina (AB176954) (>98%), had an abundance of 2.1% in BW and 1.4% in BL but was not detected in BC. P. xanthomarina has been reported as a denitrifying bacterium isolated from marine ascidians Psychrobacter namhaensis (AY722805) (>99%) and Acinetobacter johnsonii (HQ650820) (>98%), respectively. The proportion of B-OTU2 in BC and BL was 25.8% and 62.3%, and the proportion of B-OTU18 in BC and BL was 1.5% and 17.4%, respectively. However, 40.0% and 72.7% unique OTUs were found in BC and BL libraries, respectively. These results indicated that each production well appears to have a unique bacterial community despite water flooding with the same injection water for over 30 years.There were 66 bacterial OTUs (labeled as B-OTU) in these three samples . B-OTU16Methanomethylovorans (80.6%), Methanolobus (13.3%), Methanothermobacter (2.0%) and Methanobacterium (1.0%). All of the sequences in AC were affiliated with Methanobacteriaceae (93.8%) and Methanosarcinaceae (6.2%)  and Archaeoglobaceae (28.6%) (Geoglobus (28.6%) and Methanothermobacter (34.3%).Sequences of three archaeal 16S rRNA gene clone libraries  were analyzed using the same methods used for bacteria. All archaeal sequences belong to Euryarchaeota and were clustered into 4 classes of Methanomicrobia, Methanobacteria, Archaeoglobi and Thermoprotei . In AW, e (6.2%) , of whic (28.6%) . At the Methanomethylovorans thermophila (AY672821), were found in both AW and AC. A-OTU3 had an abundance of 4.0% in AW and 3.1% in AC, and A-OTU26 had an abundance of 57.6% in AW and 3.1% in AC. Other two OTUs (A-OTU16 and A-OTU20), which were related to the classes Methanomicrobia and Methanobacteria respectively, were found in both AW and AL. A-OTU16 had an abundance of 1.0% in AW and 14.3% in AL, and A-OTU20 had an abundance of 1.0% in AW and 5.7% in AL. On the other hand, A-OTU1, related to Methanobacteria, was found in all three samples and had an abundance of 2.0%, 90.6% and 22.9% in AW, AC and AL, respectively. Except for the OTUs described above, 70.6% of OTUs in AW were not detected in either production water. The comparison of archaeal communities showed that A-OTU1 and A-OTU2 were shared by two production waters, of which A-OTU2 presented in production waters only and their abundances in AC and AL were 3.1% and 2.8%, respectively. Additionally, 50% and 83.3% of the total OTUs in AC and AL were unique to themselves, indicating that each production well appears to have a specific archaeal community.There were 26 archaeal OTUs total in these three samples . Two OTUThe water flooding method has been widely used to enhance oil recovery, during which the reservoir may be influenced by the microorganisms introduced with the large volumes of injection water et al. analyzed 15 isolates from the Samotlor oil field and found 4 of them belong to new species et al. found that 16.7% of bacterial phylotypes and 50% of archaeal phylotypes detected in a high-temperature offshore oil field showed less than 97% sequence similarity with known sequences Our results have indicated that there were a large number of unexplored microorganisms in the subsurface of the Gudao oil field. Clone library analysis showed that 44.9% bacterial sequences and 20% archaeal sequences had less than 97% identity with reference sequences in the database, which seems to be a common phenomenon in petroleum reservoirs. For example, Bonch-Osmolovskaya et al. found that the similarity of the microbial structure between an injection well and a production well was lower than that of between two production wells et al. indicated that injection water contained more diverse bacteria and archaea compared with production water 2, and acetate, was dominant in the injection water. Methanobacteriales, which prefers H2 as substrates for methanogenesis, was predominant in the production waters. These results indicate that both microbial structure and metabolism are different between the injection and production wells.The microbial compositions of injection water and production waters were different according to comparison of the community structure of the three samples. Previously, using PCR-DGGE, She One reason for the different microbial structures of the injection and production wells might be the distinct difference of temperature, dissolved oxygen, and nutrient availability between the ground water recirculation system and subsurface oil strata Our results also show that a high proportion of unique bacterial and archaeal OTUs exist in the two production waters. The specificities of the two production wells are probably due to their physical and geological properties Pseudomonas. The existence of Pseudomonas in the thermophilic temperature oil reservoirs were also reported in several previous literatures, such as the Huabei Oil field with in situ temperature 75°C Psychrobacter existed in the production wells of the Gudao petroleum reservoir. Strains of Psychrobacter were previously known to be either psychrophilic, inhabiting in cold environments, such as the Antarctic or the Arctic Psychrobacter in such a high-temperature oil reservoir are still unclear and needs more investigation. The finding that bacteria of this genus degrading long chain n-alkanes Psychrobacter acting as active members in the high temperature petroleum reservoir.Additionally, many sequences obtained from two production wells in this study were affiliated with In conclusion, we demonstrated that the microbes in the production water are not significantly related with injected microorganisms and that microbial communities in adjacent production wells were different. These results prompted us to emphasize the importance of monitoring the microorganisms in production water and to understand the effects of the recovery process on the subsurface microbial ecosystem.We obtained permission from the Institute of Oil Recovery Research (Shengli Oil Field Ltd.) for observation and field studies in Gudao Oil Field which has been water flooded since 1974. The re-injection water produced from oil-water separation of production fluids, which collected from production wells in one working block. One water station supplied all the water needed for injection in the working block with several different pump stations to maintain the injection pressure. One injection well (G1–6) and two production wells (3C15 and 6–13) were used in this study. An injection water sample (W) was obtained from the water supply pipelines of G1–6, and production waters (C and L) were oil-water mixtures collected directly from the wellheads of 3C15 and 6–13, respectively. The distances between these three wells ranged from 0.3 to 0.6 km. Both injection water and production waters were full filled in 5 liters sterilized plastic bottles to prevent the diffusion of oxygen into the samples. The plastic bottles were transported to the laboratory at ambient temperature and maintained at 4°C until subsequent analysis.Demulsification of production waters was completed by adding 1/4 volume of saturated NaCl solution and heating at 70°C for 2 min. The microbial biomass was collected from 250 ml of the oil-water mixture using successive filtration with 8-µm and 0.22-µm filters . Genomic DNA was extracted from the filters using a proteinase K/sodium dodecyl sulfate (SDS)/bead beater treatment et al. The bacterial V3 region of the 16S rRNA gene was amplified using the primers P2 and P3 described by Muyzer A 250-ng aliquot of each V3 PCR product was separated on 8% (wt/vol) denatured polyacrylamide gels using a Dcode System apparatus ; the linear denaturant gradient was 27%–55% for bacteria and 30%–70% for archaea (100% denaturant corresponded to 7 M urea and 40% deionized formamide). Electrophoresis was performed in 1× Tris-acetate-EDTA (TAE) buffer at a constant voltage of 200 V and a temperature of 60°C for 240 min. The DNA bands were stained using SYBR green I  and photographed using a UV gel documentation system . The PCR-DGGE DNA profiles were digitized, and similarities between samples were analyzed using Quantity One software. Dendrograms were constructed using the unweighted pair group method using arithmetic averages (UPGMA).Bacterial and archaeal 16S rRNA gene clone libraries were constructed to examine the microbial communities of injection water and production waters. Universal primers 27F and 1492R Escherichia coli DH5α cells . Cells were spread on Luria-Bertani agar plates containing 25 µl ampicillin (50 mg/ml), 25 µl X-Gal (0.05 g/ml) and 100 µl IPTG (0.1 M).The purified PCR products were ligated into a T-vector using the pGEM-T easy vector system I according to the manufacturer's instructions (Promega). The ligation products were transformed into competent http://greengenes.lbl.gov/cgi-bin/nph-index.cgi). Operational taxonomic units (OTUs) were defined based on 97% similarity using the DOTUR program (http://www.plantpath.wisc.edu/fac/joh/dotur/documentation.html). All sequences were submitted to GenBank for analysis using the BLAST program From two clone libraries, 360 bacterial and 166 archaeal white clones were randomly selected for sequencing using an ABI 3730 DNA sequencer. Sequences were assembled after all clones were sequenced in both direction using the primers 27F/1492R for bacteria and 46F/1017R for archaea. Chimeras were detected within the sequence data set using the Chimera Check program  and the Simpson (1-D) index, were calculated using Past The GenBank accession numbers for bacterial 16S rRNA gene sequences are FJ900823-FJ901182 and HQ658567-HQ658470; the archaeal 16S rRNA gene sequences are FJ900660-FJ900822 and HQ658471.


A Tissue Microarray was constructed using immunohistochemistry to quantify expression of the HER family, E-cadherins, and podoplanin. Clinical and histopathological factors related to lymph node metastasis and prognosis were also established. Primary tumor positivity was 25.5% for EGFR, 87.3% for HER-3, and 48.1% for HER-4. Metastases were positive for EGFR in 41.7%, for HER-3 in 83.3%, and HER-4 in 43.5%. HER-2 was negative in all samples. Membrane E-cadherin and cytoplasmic E-cadherin were positive in 47.3% and 30.2% of primary tumors and 45.5% and 27.3% of metastases, respectively. Podoplanin was positive in 41.8% of primary tumors and 41.7% of metastases. Intratumoral lymphocytic infiltrate was associated with lymph node metastasis. Patients with T3 tumors had better cancer-specific survival (CSS) than those with T4 tumors; patients with no lymph node involvement had better CSS than patients with N1 tumors. Undifferentiated tumors and hyperexpression of podoplanin were negative prognostic indicators on multivariate analysis. 55 patients with advanced cutaneous squamous cell carcinoma (CSCC) of the trunk and extremities were studied.  This study identified prognostic factors including podoplanin, a novel molecular marker.”Cutaneous squamous cell carcinoma (CSCC) has a high incidence worldwide particularly in the sun exposed skin of Caucasians –5. The mE-cadherin is a transmembrane glycoprotein, and it is a mediator of calcium-dependent cell-cell adhesion in normal cells . Reduced Podoplanin is a membrane protein found on lymphatic vessel endothelium. Its function is poorly understood although it may govern endothelial motility, and its absence in animal studies is associated with lymphedema and malformation of lymphatic vessels . The aimA retrospective study of patients with locally advanced (American Joint Committee on Cancer staging T3 and T4 ) CSCC of the trunk and extremities admitted to two cancer institutions in Brazil  between 1997 and 2006 was performed. Only those patients with tumor paraffin blocks available for analysis were included. Patients with tumor infiltration of the head and neck or genital area and those with a previous cancer diagnosis other than cutaneous basal cell carcinoma were excluded. This was to avoid difficulties in identifying origin of metastasis and cause of death. 55 consecutive patients admitted and treated from October 1997 to March 2006 with a pathologic diagnosis of squamous cell carcinoma were evaluated. Patients had to have stage T3 (tumor >5 cm) or T4  tumors according to the 2002 American Joint Committee on Cancer (AJCC) staging system. Institutional Review Board approval was obtained and all clinical information retrospectively collected from medical records. Demographic and clinical variables assessed included age, gender, ethnicity, previous chronic skin lesions  at the site of the tumor, patient residence , anatomic site, and treatment. Patterns of lymph node metastases, recurrence, and survival outcomes were also recorded. Lymph node metastasis was classified as follow: N0: patients with no evidence of lymph node metastasis at presentation; N1: patients with lymph node metastasis at presentation. We considered lymph node metastasis at presentation (N1) or recurrence as the endpoint for risk of lymph node metastasis. The endpoint for survival was death from cancer. Only clinically involved lymph nodes were removed and no elective or sentinel node dissections were performed. 2, deepest tumor diameter (Breslow depth), tumor grade I to III as previously described [55 primary tumors and 22 lymph node metastases were available for pathological review by two pathologists. The pathological variables analyzed were number of mitosis/mmescribed , perineuAfter pathological review, the most representative tumor area in the paraffin block was selected for creation of a tissue microarray (TMA). Both primary tumors and lymph node metastases were selected. A Manual Tissue Arrayer I,  was used to obtain two cylinders of 1.0 millimeter in diameter from each paraffin bloc. These were implanted into the receptor paraffin block (TMA). Fifty slides were obtained and numbered from the TMA. For sample quality analysis slide numbers 1, 25, and 50 were stained with hematoxilin and eosin, and the most representative was chosen, and the other slides studied were subsequent to this. 3 for 30 minutes at 37°C and for 16 hours at 4°C.Deparaffinization of the sections was done with xylene for 15 minutes at 60°C, followed by 15 minutes at room temperature. The sections were then washed 3 times for 30 seconds with 100%, 95%, 80%, and 70% ethanol before washing in water. Endogenous peroxidase was blocked by incubating the sections in 6% hydrogen peroxide in methanol. The sections were then washed with phosphate buffered saline (PBS) 10 mM pH 7.4 for 5 minutes. Incubation followed, as described by Hsu and Raine  with theFor EGFR the H11 clone (DAKO) was used, diluted 1 : 100 in an autoclave with EDTA at pH 8.0. The following HER family polyclonal antigens were used all with citrate at pH 6.0: HER-2 (DAKO) diluted 11500 in a moist chamber, HER-3 (Neomarkers) 1 : 100 in an autoclave, (HER-4) (Neomarkers) 1 : 300 in an autoclave. The E-cadherin NCH-38 (DAKO) monoclonal antigen was diluted 1 : 600 in a moist chamber with EDTA/TRIS at pH 9.0. The Podoplanin D2–40 clone (DAKO) was used at a dilution of 1 : 200 in a moist chamber with EDTA/TRIS at pH 9.0. After incubation they were washed with PBS 3 times for 5 minutes each and then antigen amplification was performed. After amplification they were again washed with PBS 3 times for 5 minutes each. Reactions were visualized with 0.6 mg/mL 3′-3′ diaminobencidine tetrahydrochloride and 0.06% hydrogen peroxide in a 1% PBS solution for 5 minutes at 37°C. The final reaction was a brown color deposit in the cell area where the antigen-antibody reaction had occurred. The EGFR, HER-3, and HER-4 expressions were evaluated semiquantitatively according to the method described by Lager et al. . Areas wHER-2 expression was evaluated semiquantitatively according to the intensity of reaction in the cytoplasmatic membrane. Negative or weak reaction in less than 10% of cells was classified as “0”, weak reaction in more than 10% as “+”, moderate reaction in more than 10% as “++”, and strong reaction in more than 10% as “+++” positivity. For the analysis, 0 and + were considered negative and ++ and +++ positive . Cytoplasmic and membranous E-cadherin immunoexpressions were semiquantitatively evaluated as negative if the reaction occurred in up to 50% of cells and positive if occurred in more than 50% of cells.  The podoplanin (D2–40) immunoreactivity was semiquantitatively evaluated as described by Padgett et al. : negativt-test were used. Specific Cancer Survival (SCS) was also studied, and curves were constructed using the Kaplan-Meier method and compared using the univariate log-rank test. All tests were two sided, and a P-value of ≤.05 was considered statistically significant. Simultaneous prognostic effect of various factors was determined in a multivariate analysis by use of the Cox proportional-hazards regression model with a covariate of primary interest and adjustment covariates.To analyze the association between clinical variables and lymph node metastasis the chi square, Fisher exact, and 55 patients with a Mean age of 63 years (Range 30–91) were included in the study. 2 was 3 and median Breslow depth was 8 mm. Tumor characteristics are shown in The median number of mitosis/mmEGFR positivity was 25.5% in the primary tumor and 41.7% in the metastases. HER-2 was negative in all samples. HER-3 and HER-4 positivity was 87.3% and 48.1% in the primary tumor and 83.3% and 43.5% in the metastases, respectively. Membrane E-cadherin positivity was 47.3% in the primary tumor and 27.3% in the metastases. Primary tumor cytoplasmic E-cadherin was positive in 30.2% and 45.5% in the metastasis. The E-cadherin membrane/cytoplasmic ratio was 1.56 in the primary tumor and 0.60 in the metastases. Podoplanin positivity was 41.8% in primary tumor and 41.7% in metastases. p = 0.046)  . The mean and median followup was 9.6 (SD 25.0) and 25.0 months, respectively. At last followup, 19 patients were alive with no evidence of disease (34.5%), one was alive with disease (1.8%), 19 were dead of disease (34.5%), 9 dead from other causes (16.4%), and 7 lost to followup (12.7%). Those lost to followup had a mean and median followup of 24.5 (SD 21.8) and 22.1 months respectively. Only two patients were followed up for less than one year and three patients for less than 22 months. For the 11 patients that presented with lymph node metastasis during the followup, median time to occurrence was 13.08 months. p = 0  .001). Patients with no lymph node metastases had a 5-year CSS of 63.3%, with no 5 year survivors in patients with lymph node metastases (p = 0.004). Gender, race, ambient, anatomic location, location of metastasis, and presence of previous nononcologic lesions did not affect survival . No significant difference in 5-year CSS was seen for Breslow depth, peritumoral lymphocytic infiltrate, vascular, and perineural infiltration. The only histological variable that had a significant impact on survival was the tumor grade. Patients with grade I lesions had a 5-year CSS of 82.2% compared with 23.8% in patients with grade II and III tumors (p = 0  .010). Podoplanin negative patients had a higher 5 years (p = 0  .038). Clinical stage and Podoplanin positivity were independent prognostic factors on both univariate and multivariate analysis  was 49.7%. For patients with T3 tumors 5-year CSS was 67.6%, and no patients with T4 tumors were alive at 5 years  had no impact on lymph node metastasis and did not influence survival. Breslow depth may correlate with lymph node metastasis and survival in less advanced tumors, but in this study median tumor thickness was high which may have weakened any association. Low-grade tumors were associated with prolonged survival confirming the aggressive nature of undifferentiated tumors. Unlike squamous cell carcinoma of the head and neck or esophagus, EGFR had no influence on prognosis. It is possible that altered EGFR expression may be associated with local recurrence, which is more frequently life threatening at other sites. HER-2 was negative in all samples and may play little part in CSCC progression as found in squamous cell carcinomas from other sites. High HER-4 expression in lymph node metastases was associated with poor prognosis suggesting a role in progression of CSCC of the trunk and extremities. It is possible that altered HER-4 expression occurs late and is present only in metastases. The altered coexpression of the HER family may play a role  but the small number of cases in this study meant this could not be analyzed. E-cadherin expression had no significant association with lymph node metastasis or survival, but the expression ratio between membrane and cytoplasm was lower in the metastasis, suggesting accumulation with a loss of function. This may be due to a mutated E-cadherin resulting in cytoplasmic accumulation with a loss of cell adhesion and disease progression , 34. SomPodoplanin expression was not associated with the presence of lymph node metastasis, but was a prognosticator of reduced survival indicating a locally aggressive tumor, with survival impact. Altered expression of podoplanin is associated with mesothelioma, squamous cell carcinoma of oral mucosa, and germ cell tumors, suggesting that podoplanin may influence invasive and proliferative activity –40. As CPatients with advanced CSCC of trunk and extremities with poor prognostic factors such as undifferentiated, T4, N1 tumors, high podoplanin expression in the primary tumor, or high HER-4 expression in the lymph node metastasis may be candidates for new more aggressive modalities of treatment. Further studies using these molecular markers are needed to help refine treatment of CSCC of the trunk and extremities.


Ag/AgCl) using an electrochemical cell containing aniline and polyvinylsulphonate. Firstly, the optimum working conditions for preparing polyaniline-polyvinylsulfonate films were investigated. Determination of glucose was carried out by the oxidation of enzymatically produced H2O2 at 0.4 V vs. Ag/AgCl. The effects of pH and temperature were investigated and the optimum pH value was found to be 7.5. The storage stability and operational stability of the enzyme electrode were also studied. The results show that 75% of the response current was retained after 16 activity assays. The prepared glucose biosensor retained 80.6% of initial activity after 40 days when stored in 0.1 M phosphate buffer solution at 4 °C.In this study, a novel amperometric glucose biosensor with immobilization of glucose oxidase on electrochemically polymerized polyaniline-polyvinylsulphonate (Pani-Pvs) films has been accomplished via the entrapment technique. Electropolymerization of aniline on the Pt surface of the Pt electrode was carried out at constant potential (0.75 V,  After entrapping the glucose oxidase onto the Pani-Pvs film, the electrode was rinsed with deionized water to remove the unreacted aniline monomer and free glucose oxidase. Immobilized enzyme electrodes were kept in a refrigerator at the 4 °C in phosphate buffer when not in use.Immobilization of glucose oxidase was carried out by the physical entrapment technique. Pt/Pani-Pvs electrode was immersed in 0.2 M aniline and 2.5 mL aqueous solution of the sodium salt of polyvinylsulphonate (25%). Then 1 mL of GOX enzyme (10 U/mL) was added into the solution, which was purged with argon for the removal of oxygen before electropolymerization. The polymerization was carried out at constant potential of 0.75 V for 60 min [2.5.vs. Ag/AgCl electrode (3 M KCl)] by detecting the H2O2 oxidation current. After the background current reached a stable value, glucose solution was added to the cell using a micropipette and it was stirred. Then the resulting current difference was recorded. Research into the operational and storage stability as well as effects of pH and temperature were carried out using 1.0 × 10−5 M concentrations of glucose.Amperometric response studies carried out in phosphate buffer (0.1 M pH 7.5). Operational stability, storage stability, pH and temperature were determined via application of 0.4 V −1/Δi graph (Lineweaver-Burk plot)  . Km valu 9.34 mM ,16. This3.9.−5 M). At the end of the 16 measurements, the biosensor had lost 25% of its initial activity  16 times in the same day at constant temperature, pH and substrate concentration  and uric acid (in 3.0 × 10−4 M) on the analysis of glucose were found to be 15% and 25%, respectively. These interferences were almost removed by dilution of solutions in the cell.A few common substances found in serum or urine were studied for any interfering effects on the analysis of glucose. Known concentrations of ascorbic acid, uric acid and paracetamol (acetaminophen) were added. It was observed that a paracetamol concentration of 1.0 × 104.In this study, glucose oxidase was successfully immobilized on a poly (aniline)-polyvinylsulphonate (Pani-Pvs) composite film. The experimental results clearly showed that the resulting biosensor exhibited good performance in the determination of glucose. It was seen that glucose biosensor was sensitive and its operational stability and long term storage stability were found to be good.