Search

We recently establish that aspafilioside B, a steroidal saponin extracted from Asparagus filicinus, is an active cytotoxic component. However, its antitumor activity is till unknown. In this study, the anticancer effect of aspafilioside B against HCC cells and the underlying mechanisms were investigated. Our results showed that aspafilioside B inhibited the growth and proliferation of HCC cell lines. Further study revealed that aspafilioside B could significantly induce G2 phase cell cycle arrest and apoptosis, accompanying the accumulation of reactive oxygen species (ROS), but blocking ROS generation with N-acetyl-l-cysteine (NAC) could not prevent G2/M arrest and apoptosis. Additionally, treatment with aspafilioside B induced phosphorylation of extracellular signal-regulated kinase (ERK) and p38 MAP kinase. Moreover, both ERK inhibitor PD98059 and p38 inhibitor SB203580 almost abolished the G2/M phase arrest and apoptosis induced by aspafilioside B, and reversed the expression of cell cycle- and apoptosis-related proteins. We also found that aspafilioside B treatment increased both Ras and Raf activation, and transfection of cells with H-Ras and N-Ras shRNA almost attenuated aspafilioside B-induced G2 phase arrest and apoptosis as well as the ERK and p38 activation. Finally, in vivo, aspafilioside B suppressed tumor growth in mouse xenograft models, and the mechanism was the same as in vitro study. Collectively, these findings indicated that aspafilioside B may up-regulate H-Ras and N-Ras, causing c-Raf phosphorylation, and lead to ERK and p38 activation, which consequently induced the G2 phase arrest and apoptosis. This study provides the evidence that aspafilioside B is a promising therapeutic agent against HCC., (© 2015 Wiley Periodicals, Inc.)

There has been considerable interest in the biological functions of astaxanthin and its potential applications in the nutraceutical, cosmetics, food, and feed industries in recent years. However, the unstable structure of astaxanthin considerably limits its application. Therefore, this study reports the encapsulation of astaxanthin in calcium alginate beads using the extrusion method to improve its stability. This study also evaluates the stability of the encapsulated astaxanthin under different storage conditions. The evaluation of astaxanthin stability under various environmental factors reveals that temperature is the most influential environmental factor in astaxanthin degradation. Stability analysis shows that, regardless of the formulation used, the content of astaxanthin encapsulated in alginate beads remains above 90% of the original amount after 21 days of storage at 25°C. These results suggest that the proposed technique is a promising way to enhance the stability of other sensitive compounds.

Background: Alpha-blockers are notorious for their first-dose effect of acute hypotension during the early initiation period. Because acute cerebral hypoperfusion may precipitate an episode of ischemic stroke, we aimed to provide a quantitative estimate of the risk of ischemic stroke during the early initiation period of α-blocker therapy, using a self-controlled case series design., Methods: We identified all men aged 50 years or more as of 2007 who were incident users of α-blockers and had a diagnosis of ischemic stroke during the 2007-2009 study period using claims data from Taiwan's National Health Insurance claims database. The first day on which the α-blocker was prescribed was the index date. We partitioned different risk periods according to their relationship to the index date (pre-exposure risk periods 1 and 2 = ≤ 21 d and 22-60 d before index date, respectively; post-exposure risk periods 1 and 2 = ≤ 21 d and 22-60 d after index date, respectively); the remainder of the study period was defined as the unexposed period. We estimated the incidence rate ratio (IRR) of ischemic stroke in each risk period relative to the unexposed period using a conditional Poisson regression model., Results: A total of 7502 men were included. Compared with the risk in the unexposed period, the risk of ischemic stroke was increased in post-exposure risk period 1 among all patients in the study population (adjusted IRR 1.40, 95% confidence interval [CI], 1.22-1.61) and among patients without concomitant prescriptions for other antihypertensive agents (adjusted IRR 2.11, 95% CI 1.73-2.57)., Interpretation: Alpha-blocker therapy was associated with an increased risk of ischemic stroke during the early initiation period, especially among patients who were not taking other antihypertensive agents., (© 2016 Canadian Medical Association or its licensors.)

Background: Methamphetamine is a commonly used illicit drug that threatens many adolescents and youth in Asian Pacific countries, contributing to the heavy burden of disease and drug-related death. Little is known about the prevalence of associated psychological problems in methamphetamine-dependent adolescents and youth., Purpose: The study sought to estimate the prevalence of psychological problems in adolescents and youth in China with methamphetamine dependence and to identify factors related to co-occurring psychological problems., Methods: A cross-sectional design was used. Methamphetamine-dependent adolescents and youth from two drug rehabilitation centers in Tianjin, China, took part. Each participant was assessed with a sociodemographic questionnaire and Symptom Checklist-90 (SCL-90)., Results: A total of 161 adolescents and youth took part. Psychological symptomatology was high, with 59.6% of participants having over 160 points on the SCL-90 total score, indicating significant psychological distress. On the basis of subscale scores, phobic anxiety, depression, and anxiety were common. Male participants, younger adolescents, and those who are addicted to methamphetamine for a longer time with more consumption were more likely to experience psychological problems., Discussion: Nurses should be aware of the type and severity of psychological problems among methamphetamine-dependent adolescents and youth. Nursing care should be designed to relieve psychological problems and improve mental health of the methamphetamine-dependent adolescents and youth.

Low heart rate variability (HRV) has been recognized to correlate with adverse cardiovascular (CV) outcomes in hemodialysis (HD) patients. It has been reported that HRV might be improved after HD, but whether the improved HRV after HD predicts a better CV prognosis remains to be determined. This study examined the ability of the change in HRV before and after HD in predicting overall and CV mortality in HD patients. This study enrolled 182 patients under maintenance HD. HRV was examined to assess changes before and after HD. The change in HRV (ΔHRV) was defined as post-HD HRV minus pre-HD HRV. During a median follow-up period of 35.2 months, 29 deaths (15.9%) were recorded. Multivariate analysis showed that decreased ΔLF% was associated with increased overall (hazard ratios [HR], 0.978; 95% confidence interval [CI], 0.961-0.996; p = 0.019) and CV mortality (HR, 0.941; 95% CI, 0.914-0.970; p < 0.001), respectively. Moreover, adding ΔLF% to a clinical model provided an additional benefit in the prediction of overall (p = 0.002) and CV mortality (p < 0.001). HRV change before and after HD (ΔHRV) is an useful clinical marker, and it is stronger than HRV before HD in predicting overall and CV mortality.

Objective: Because the number of long-term survivors of colorectal cancer has increased, second primary cancer has become an important issue. However, previous studies were heterogeneous in design, and few data for Asia-Pacific area were available., Methods: This was a retrospective population-based study conducted using the national database of the Taiwan Cancer Registry. Patients who have histology-proven primary colon cancer and rectal cancer from 1995 to 2005 were enrolled in this study. All second primary cancer events had to be histology proven. The standardized incidence ratio of second primary cancer was used as an indicator. Standardized incidence ratio was counted as the number of observed second primary cancer divided by the expected number of cancer cases in the general population., Results: A total of 65 648 eligible index patients were enrolled, and 3810 second primary cancer events were identified. The standardized incidence ratio for all of the patients was 1.03 (95% confidence interval: 0.99-1.06), which implied that the risk of second primary cancer was not significantly elevated in the index patients compared with that of the general population. The standardized incidence ratio for the patients aged <50, 50-70 and >70 years was 2.52 (95% confidence interval: 2.28-2.78), 1.18 (95% confidence interval: 1.12-1.23) and 0.80 (95% confidence interval: 0.76-0.84), respectively. In young patients (aged <50 years), the standardized incidence ratio increase was statistically significant and persisted for over 10 years and this significantly increased across all subgroups. The small intestine, the large intestine, the female genital organs and the lungs were the most common sites of second primary cancer in young patients., Conclusions: Young patients with colorectal cancer have an increased risk of developing second primary cancer., (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Aims: This study aimed to evaluate the risk of hip/femur fractures during the initiation period of α-adrenoceptor blocker therapy using the National Health Insurance claims database, Taiwan, with a self-controlled case series design., Methods: All male beneficiaries aged over 50 years as of 2007, who were incident users of α-adrenoceptor blockers and also had a diagnosis of hip/femur fracture within the 2007-2009 study period were identified. The first day when the α-adrenoceptor blocker was prescribed was set as the index date. We partitioned the initial 21 day period following the index date as the post-exposure risk period 1, days 22-60 after the index date as the post-exposure risk period 2, the 21 day period prior to the index date as the pre-exposure risk period 1 and days 22-60 prior to the index date as the pre-exposure risk period 2. The remainder of the study period was defined as the unexposed period. The incidence rate ratio (IRR) of hip/femur fractures within each risk period compared with the unexposed period was estimated using a conditional Poisson regression model., Results: A total of 5875 men were included. Compared with the unexposed period, the IRR of hip/femur fractures was 1.36 (95% confidence interval 1.06, 1.74, P = 0.017) within the post-exposure risk period 1 for patients without concomitant prescriptions of anti-hypertensive agents., Conclusions: Use of α-adrenoceptor blockers was associated with a small but significant increase in the risk of hip/femur fractures during the early initiation period in patients without concomitant prescriptions of anti-hypertensive agents., (© 2015 The British Pharmacological Society.)

The large yellow croaker Larimichthys crocea is an important mariculture fish species in China, and the bacterium Vibrio harveyi (V. harveyi) and the ciliate protozoan Cryptocaryon irritans (C. irritans) are the two major pathogens in its aquaculture sector. Interferon-gamma (IFN-γ) plays important roles in regulating both innate and cell mediated immune responses as an inflammatory cytokine. In this study, we obtained the nucleotide sequence of IFN-γ from the large yellow croaker (LcIFN-γ). The phylogenetic relationship tree of 18 available IFN-γ genes was constructed based on their sequences. Expression analyses in 10 various tissues were conducted after the croaker challenged with V. harveyi and C. irritans, respectively. Real time PCR analysis showed that the expression of LcIFN-γ was observed broadly in health individuals. After injected with V. harveyi, the 10 tissues had a higher expression of IFN-γ at the first day (1 d); only spleen, muscle, intestine, heart and skin had higher expressions after infected with C. irritans at 1 d. Major immune tissues (skin, gill, head kidney and spleen) and detoxification tissue (liver) were sampled at 0 h, 6 h, 1 d, 2 d, 3 d, 4 d, 5 d and 7 d to understand the expression trends of LcIFN-γ after challenged with C. irritans. The expressions of LcIFN-γ in skin and gill (the primary immune organs) showed a clear correlative relationship with the life cycle of C. irritans., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Given the growing number of arthritis patients and the limitations of current treatments, there is great urgency to explore cartilage substitutes by tissue engineering. In this study, we developed a novel decellularization method for menisci to prepare acellular extracellular matrix (ECM) scaffolds with minimal adverse effects on the ECM. Among all the acid treatments, formic acid treatment removed most of the cellular contents and preserved the highest ECM contents in the decellularized porcine menisci. Compared with fresh porcine menisci, the content of DNA decreased to 4.10%±0.03%, and there was no significant damage to glycosaminoglycan (GAG) or collagen. Histological staining also confirmed the presence of ECM and the absence of cellularity. In addition, a highly hydrophilic scaffold with three-dimensional interconnected porous structure was fabricated from decellularized menisci tissue. Human chondrocytes showed enhanced cell proliferation and synthesis of chondrocyte ECM including type II collagen and GAG when cultured in this acellular scaffold. Moreover, the scaffold effectively supported chondrogenesis of human bone marrow-derived mesenchymal stem cells. Finally, in vivo implantation was conducted in rats to assess the biocompatibility of the scaffolds. No significant inflammatory response was observed. The acellular ECM scaffold provided a native environment for cells with diverse physiological functions to promote cell proliferation and new tissue formation. This study reported a novel way to prepare decellularized meniscus tissue and demonstrated the potential as scaffolds to support cartilage repair.

Background: The course of incident delirium and subsyndromal delirium (SSD) after cardiac surgery is not well studied., Objective: The aim of this study was to evaluate the course of incident delirium and SSD, their risk factors, and impact on patients' cognitive function after elective coronary artery bypass graft (CABG) surgery., Methods: Consecutive patients scheduled for an isolated CABG at a tertiary medical center in Taiwan were enrolled if they had no preoperative delirium symptoms. Delirium was assessed daily for 1 week after surgery using the Confusion Assessment Method. Subsyndromal delirium was defined as presenting with any core symptom below the diagnostic threshold for delirium. Cognitive function was assessed by the Mini-mental State Examination., Results: Of 38 participants, 7 had incident (first-time) delirium (18.4% incidence) and 13 had incident SSD (34.2% incidence). Whereas SSD usually lasted 1 day, delirium changed gradually to SSD to recovery and its symptomatology lasted longer. We identified 6 delirium risk factors: older age, more comorbidities, cardiac pulmonary bypass, blood transfusion, larger transfusion volume, and longer duration of intraoperative blood pressure less than 60 mm Hg. The frequencies of these risk factors for SSD were often intermediate between those of risk factors in groups with and without delirium. By hospital discharge, participants with delirium had the longest hospital stays and lowest cognitive scores, those with SSD had intermediate stays and scores, and those without delirium had the lowest stays and scores., Conclusion: Delirium and SSD after CABG are common. Greater number and severity of risk factors for delirium may predict increasingly poor outcomes, with the dose-response relationship between risk factors and outcomes for SSD intermediate between that for no symptoms and full delirium. Intervention trials are indicated, particularly for patients with a greater number and severity of predisposing and precipitating risk factors.

Objective: To assess the relationship between smoking status and health-related quality of life 1 year after participation in a smoking cessation programme in Taiwan., Design: A cohort study of smokers who voluntarily participated in a smoking cessation programme with two follow-up assessments of smoking status via telephone interview, conducted 6 months and 1 year after finishing the smoking cessation programme., Setting: Hospitals and clinics providing smoking cessation services., Participants: A total of 3514 participants completed both telephone interviews, which represents a response rate of 64%. After the interviews, participants were divided into four groups according to their smoking status: (1) long-term quitters: participants who had quit tobacco use for 1 year; (2) short-term quitters: participants who had been smoking for at least 6 months and then quit tobacco for 6 months after participating in the programme; (3) relapsed smokers: participants who relapsed into tobacco use after ceasing tobacco use for 6 months; and (4) continuing smokers: participants who failed to quit smoking for at least 1 year, despite participating in the programme., Interventions: The Outpatient Smoking Cessation Service of Taiwan provides counselling and pharmacotherapy to individuals seeking to quit smoking., Primary Outcomes: The health-related quality of life of the participants was measured using an approved Chinese version of the EuroQol-5D-3L (EQ-5D-3L) descriptive system., Results: After controlling for sex, age, education, marital status, job status, monthly income and disease status at baseline, our results revealed that long-term (OR=0.61 (0.48 to 0.77)) and short-term (OR=0.65 (0.54 to 0.79)) quitters experienced less anxiety and depression than did continuing smokers., Conclusions: Our study provides evidence to support claims that all quitters, regardless of whether they stop smoking for 6 months or 1 year, have better quality of life with regard to anxiety or depression., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.)

The clinical outcomes of different limus-based drug-eluting stents (DES) in a real-world setting have not been well defined. The aim of this study was to investigate the clinical outcomes of three different limus-based DES, namely sirolimus-eluting stent (SES), Endeavor zotarolimus-eluting stent (E-ZES) and everolimus-eluting stent (EES), using a national insurance claims database. We identified all patients who received implantation of single SES, E-ZES or EES between January 1, 2007 and December 31, 2009 from the National Health Insurance claims database, Taiwan. Follow-up was through December 31, 2011 for all selected clinical outcomes. The primary end-point was all-cause mortality. Secondary end-points included acute coronary events, heart failure needing hospitalization, and cerebrovascular disease. Cox regression model adjusting for baseline characteristics was used to compare the relative risks of different outcomes among the three different limus-based DES. Totally, 6584 patients were evaluated (n=2142 for SES, n=3445 for E-ZES, and n=997 for EES). After adjusting for baseline characteristics, we found no statistically significant difference in the risk of all-cause mortality in three DES groups (adjusted hazard ratio [HR]: 1.14, 95% confidence interval [CI]: 0.94-1.38, p=0.20 in E-ZES group compared with SES group; adjusted HR: 0.77, 95% CI: 0.54-1.10, p=0.15 in EES group compared with SES group). Similarly, we found no difference in the three stent groups in risks of acute coronary events, heart failure needing hospitalization, and cerebrovascular disease. In conclusion, we observed no difference in all-cause mortality, acute coronary events, heart failure needing hospitalization, and cerebrovascular disease in patients treated with SES, E-ZES, and EES in a real-world population-based setting in Taiwan.

Colonoscopy remains the gold standard for colorectal cancer screening. Many barriers to the procedure exist including the possibility of abdominal discomfort that may occur with insufflation. Carbon dioxide (CO2), which is rapidly absorbed in the blood stream, is an alternate method used to distend the lumen during colonoscopy. The goal of this study was to compare patient discomfort, abdominal girth, and recovery time in 2 groups of patients randomized to CO2 versus room air insufflation during colonoscopy. Using a Wong-Baker score, we found statistical difference in postprocedural discomfort levels (CO2 Group: 1.15 ± 2.0 vs. room air: 0.41 ± 0.31, p = .015) and a significantly greater increase in abdominal girth over CO2 immediately postprocedure (room air: 1.06 ± 1.29 inches vs. CO2: 0.56 ± 0.73 inches, p = .054) girth immediately postprocedure; however, recovery time was similar between the 2 study arms (CO2: 9.1 ± 16.2 minutes vs. room air: 10.2 ± 18.6 minutes, p = .713). Further studies are needed to determine whether CO2 is cost-effective and improves patient satisfaction with colonoscopy.

Background: Randomized trials suggest that radiofrequency ablation (RFA) may be more effective than percutaneous ethanol injection (PEI) in the treatment of hepatocellular carcinoma (HCC). However, the survival advantage of RFA needs confirmation in daily practice., Methods: We conducted a population-based cohort study using the Taiwan Cancer Registry, National Health Insurance claim database and National Death Registry data from 2004 through 2009. Patients receiving PEI or RFA as first-line treatment for newly-diagnosed stage I-II HCC were enrolled., Results: A total of 658 patients receiving RFA and 378 patients receiving PEI treatment were included for final analysis. The overall survival (OS) rates of patients in the RFA and PEI groups at 5-year were 55% and 42%, respectively (p < 0.01). Compared to patients that received PEI, those that received RFA had lower risks of overall mortality and first-line treatment failure (FTF), with adjusted hazard ratios (HRs) [95% confidence interval (CI)] of 0.60 (0.50-0.73) for OS and 0.54 (0.46-0.64) for FTF. The favorable outcomes for the RFA group were consistently significant for patients with tumors > 2 cm as well as for those with tumors < 2 cm. Consistent results were also observed in other subgroup analyses defined by gender, age, tumor stage, and co-morbidity status., Conclusion: RFA provides better survival benefits than PEI for patients with unresectable stage I-II HCC, irrespective of tumors > 2 cm or ≤ 2 cm, in contemporary clinical practice.

Curcumin is extracted from the rhizomes of the ginger family plant Curcuma longa L., which has a good protection for liver, kidney, and immune system. However, there is little information about its contribution in protection of astrocytes recently. The present study was undertaken to elucidate the protective effect of curcumin, an herbal antioxidant, on 1-methyl-4-phenylpyridinium ion- (MPP(+)-) and lipopolysaccharide- (LPS-) induced cytotoxicities, as well as the underlying mechanisms by using primary mouse mesencephalic astrocytes. The results showed that curcumin protected the mesencephalic astrocytes from MPP(+)- and LPS-induced toxicities along with reducing reactive oxygen species (P < 0.05) and maleic dialdehyde (P < 0.05) sufficiently. Moreover, curcumin significantly inhibited the cytochrome P450 2E1 (CYP2E1) expression (P < 0.01 at mRNA level, P < 0.05 at protein level) and its activity (P < 0.05) sufficiently induced by MPP(+) and LPS in the mouse mesencephalic astrocytes. And curcumin as well as diallyl sulphide, a CYP2E1 positive inhibitor, ameliorated MPP(+)- and LPS-induced mouse mesencephalic astrocytes damage. Accordingly, curcumin protects against MPP(+)- and LPS-induced cytotoxicities in the mouse mesencephalic astrocyte via inhibiting the CYP2E1 expression and activity.

Aims: In this study, we investigated the peripheral mechanisms underlying the metabolic side effects of fluoxetine (FLX) by focusing on hepatic lipid metabolism., Methods: Primary mouse hepatocytes were prepared from male mice by the two-step perfusion method. The lipid accumulation in primary mouse hepatocytes was analyzed via neutral oil staining. And the lipid metabolism enzymes were determined with RT-PCR and Western blot., Results: Fluoxetine significantly induced the lipid accumulation in primary mouse hepatocytes. Moreover, FLX increased the acetyl-CoA carboxylase 1 (ACC1) and fatty acid synthase (FAS) expression, which are important enzymes in lipogenesis. Oppositely, Fluoxetine significantly decreased the carboxylesterase 3 (CES3) and carboxylesterase 1 (CES1) expression, which are related to lipolysis. Further study demonstrated FLX-activated SREBP1c, which is one of the most important transcription factors conducting coordinated transcriptional regulation of lipogenesis gene such as ACC1 and FAS. And the increase of lipogenesis gene (ACC1) was abolished by SB203580 but not by pyrrolidine dithiocarbamate (PDTC), suggesting through p38-MAPK pathway., Conclusion: Fluoxetine induces hepatic lipid accumulation via both promotion of the SREBP1c-related lipogenesis and reduction of lipolysis in primary mouse hepatocytes., (© 2012 Blackwell Publishing Ltd.)

8-Methoxypsoralen (8-MOP), a naturally occurring compound, is a potent modulator of epidermal cell growth and differentiation in combination with ultraviolet light. However, there is little information on 8-MOP contribution to cell apoptosis alone. In the study, we evaluated 8-MOP, independently of its photoactivation, induced apoptosis in human hepatocellular carcinoma HepG2 cells. And we provide a molecular explanation linking 8-MOP to induce apoptosis. In HepG2 cells, treatment with 8-MOP induced the cell apoptosis in both dose-dependent and time-dependent manners. IC(50) values of 8-MOP were 8.775, 5.398 μM for 48 and 72 h, respectively. Further study showed that 8-MOP decreased the procaspase-3, procaspase-8, and procaspase-9, increased the ratio of Bax/Bcl-2 and decreased the survivin. Moreover, 8-MOP decreased differentiated embryonic chondrocyte gene1 (DEC1). Overexpression of DEC1 antagonized partially apoptosis induced by 8-MOP. And overexpression of DEC1 abolished the decrease of survivin and the activation of caspase-3 induced by 8-MOP partially. So, down regulation of DEC1 is involved in 8-MOP-induced apoptosis in HepG2 cells. Here, it is demonstrated that DEC1 possesses anti-apoptotic effects in 8-MOP-treated HepG2 cells. The findings provide more of a basis for 8-MOP as an anti-tumor agent in cancer therapy., (Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.)

Objective: To observe the therapeutic effect of type II early diabetic nephropathies intervened by acupoint thread embedding for strengthening spleen and benefiting kidney., Methods: Sixty cases of type II early diabetic nephropathies were randomly divided into an acupoint thread embedding group and a routine therapy group, 30 cases in each group. In routine therapy group, western medicine routine therapy was applied to control blood sugar, blood pressure and blood lipid. In acupoint thread embedding group, besides the western medicine routine therapy, thread embedding intervention was given at Pishu (BL 20), Zusanli (ST 36), Shenshu (BL 23) and Yishu (Extra) on both sides as main acupints, and the treatment course was 3 months. Urinary micro-albumin excretion (UAER), total score of TCM syndrome and monitor control indices (blood sugar, blood pressure, blood lipid, urea nitrogen and serum creatinine) in both groups were observed before and after treatment, and the therapeutic effects in both groups were compared., Results: After treatment, the indices of UAER and total score of TCM syndrome were all reduced (P < 0.001, P < 0.01); the reduction in acupoint thread embedding group was more obvious (P < 0.01, P < 0.05); the total effective rate in acupoint thread embedding group was 76.7% (23/30), superior to that of 63.3% (19/30) in routine therapy group (P < 0.05). The blood sugar, blood pressure and blood lipid in both groups were remarkably improved (all P < 0.001) after treatment; the urea nitrogen and creatinine had no notable variation., Conclusion: Acupoint thread embedding combined with western medicine routine therapy can not only reduce the urinary micro-albumin excretion of type II early diabetic nephropathies, but also improve the Chinese medicine symptoms and the therapeutic effect is superior to that of simple western medicine routine therapy.

Background: Asian ethnicity is associated with a distinct molecular etiology, treatment response, and survival outcome among patients with non-small cell lung cancer (NSCLC). This study examines the survival impact of platinum-based adjuvant chemotherapy for Asian patients with stage I-IIIA NSCLC., Methods: This study recruited patients aged ≥18 years with histologically proven stage IA-IIIA NSCLC registered in the Taiwan Cancer Registry database in January 2004 to December 2007. Platinum-containing adjuvant chemotherapy had to be started within 90 days of the primary surgery. Kaplan-Meier survival curves, log-rank tests, and the Cox proportional hazards regression model were used to assess the influence of various risk factors on survival time., Results: This study included 2,231 patients with stage IA-IIIA NSCLC who underwent primary surgery with a clear surgical margin. The percentages of all causes of death were significantly lower for the chemotherapy group for both stage II and stage IIIA patients. Multivariate analysis identified platinum-based adjuvant chemotherapy as an independent prognostic factor for the overall survival outcome of stage II (hazard ratio [HR], 0.61; 95% confidence interval [CI], 0.39-0.94; p = .024) and IIIA (HR, 0.71; 95% CI, 0.52-0.96; p = .029) patients. Among these patients, those who received adjuvant chemotherapy had a superior overall survival outcome for both genders, for the subgroup of patients aged ≥70 years, and for those with adenocarcinoma., Conclusion: Platinum-based adjuvant chemotherapy should be considered in the treatment plan for Asian patients with resected stage II and stage IIIA NSCLC.

An oleic acid-based microemulsion system with a member of the Tween series or Cremophor EL as the surfactant and a short-chain alcohol as the cosolvent was developed for rapid-onset intranasal delivery of sildenafil. The phase behaviour and solubilization capacity of the microemulsion system were characterized, and nasal absorption of sildenafil from the microemulsion formulations was investigated in rabbits. Sildenafil displayed a high solubility of 124 mg/mL in the microemulsion consisting of 40% oleic acid, 10% H(2)O, and 50% Tween 80:ethanol (EA) (at a 1:4 weight ratio). Nasal absorption of sildenafil from this microemulsion was found to be fairly rapid. With a 10-mg dose, the onset of action was arrived instantly following intranasal administration and the duration was over 3 h using an in vivo rabbit studies. In addition, nasal ciliotoxicity studies were carried out using in vivo rat nasal mucosa model and showed no ciliotoxicity. Therefore, the prepared systems are no serious nasal ciliotoxicity for intranasal administration. The microemulsion system composed of oleic acid, Tween 80, EA, and water may be a practical approach for the rapid-onset delivery of sildenafil for the treatment of erectile dysfunction.

In order to characterize the dense and regular level of aerobic granular sludge (AGS) which was cultivated by different carbon sources, the SEM images of mature AGS were used in the study. The calculation process using Photoshop and Fips2 on fractal characteristics of these granules was built. The lowest box-counting dimension was bulking aerobic granular sludge cultivated by glucose (No. 1), which was 1.794 +/- 0.011; the box-counting dimension of AGS cultivated by peptone (No. 3) and domestic sewage (No. 5) were reached up to 1.866 +/- 0.018 and 1.880 +/- 0.015. The boundary box-counting dimension of the AGS was also calculated and the average value was 1.14. The AGS cultivated by beer (No. 6) was more regular in shape and the boundary box-counting dimension was 1.115 +/- 0.003. The AGS cultivated by landfill leachate (No. 7) was the most irregular in shape. This study indicates that fractal dimension provides an approach for quantification of dense and regular level of AGS, furthermore, it could be used to characterize the status of AGS, such as bulking.

Aerobic granular sludge (AGS) for simultaneous nitrogen and phosphorus removal (SNPR) was cultivated and studied in two lab-scale sequencing batch reactors (named as A & B) treating real domestic wastewater, additional carbon (sodium propionate and sodium acetate for A, glucose for B) was added to make the ratio of COD:N:P as 360:60:6, good SNPR was achieved at normal (18-27 degrees C) and low temperature (9-13 degrees C). The microbial community composition and distribution, distribution of cells and extracellular polymeric substances (EPS) and morphologies of the AGS were investigated using fluorescence in situ hybridization (FISH), in situ fluorescent staining and scanning electron microscope (SEM), respectively. FISH results showed that ammonium-oxidizing bacteria comprised 12% of all the bacteria and were mainly located at the outer parts of the granules; phosphates accumulating organisms comprised 40% of all the bacteria and were mainly located in the inner parts of the granules. Nitrification was the rate controlling step; denitrifying phosphate accumulating organisms inside the granular sludge might be responsible for denitrification in the aerobic phase, which enabled effective SNPR. Live/dead fluorescent staining results showed that dead cells were distributed throughout the granules and live cells were principally distribution of polysaccharide (including alpha-mannopyranosyl, alpha-glucopyranosyl sugars and beta-D-glucopyranose polysaccharides) of EPS in AGS were influenced by different carbon sources, but the contents and distributions of protein and lipids were not, the contents of protein was the largest. Polysaccharide was responsible for the formation and maintenance of aerobic granular sludge. SEM results showed that bacilli and cocci were the main bacteria in the granules of A and B, respectively. Carbon sources affected the bacteria type and the SNPR efficiency, sodium propionate and sodium acetate were better than glucose.

The thyroid hormone receptor (TR) and human testicular orphan receptor 4 (TR4) belong to the nuclear hormone receptor superfamily. They are ligand-dependent transcription factors. TR and TR4 bind to a similar thyroid response element (TRE), known as a direct repeat with four nucleotide spacing (DR4). This study examined the possible interaction or cross-talking between those two receptors. We hypothesized that protein-protein interaction between TR4 and TR may promote TR-mediated transcriptional activity. Glutathione S-transferase pull-down and immunoprecipitation assays showed direct interaction between TR and TR4. Electrophoretic mobility-shift assay demonstrated that TR and TR4 could co-occupy the same TRE. The interaction between TR4 and TR may enhance regulation of genes targeted by TR, such as furin, fibrinogen, cdk2 and p21 expression. We found that TR4 function is similar with TR as TR4 alone could regulate expression of some TR target genes, and could increase cell migration or inhibit cell proliferation. Importantly, the TR-dependent inhibition of cell proliferation and stimulation of cell migration are more enhanced in the HepG2-TR cells stably over-expressing TR4. Overall, TR4 not only has modulation abilities similar to TR but also can cross-talk with TR and promote the TR signaling pathway., ((c) 2009 Wiley-Liss, Inc.)

The aim of this study was to develop an optimal gastroretentive drug delivery system (GRDDS) for administering Losartan. Additionally, the influence of optimized GRDDS on the bioavailability of Losartan and the formation extent of active metabolite E3174 by CYP2C9 polymorphism was investigated. Swellable and floatable GRDDS tablets combining hydroxyethyl cellulose (HEC), sodium carboxymethyl cellulose (NaCMC), and sodium bicarbonate were prepared at various compression pressures for evaluating swelling characteristics and floating capacity. Then Losartan was incorporated into optimized formulations for in vitro and in vivo characterizations. An appropriate ratio of HEC to NaCMC, addition of sodium bicarbonate, and compression at lower pressures resulted in the tablets floating over SGF for more than 16 h and swelling to 2 cm in diameter within 3h. The release patterns of Losartan from these tablets were pH-dependent. Results of the clinical trials showed that the mean bioavailability from GRD-A (HEC 91.67%, sodium bicarbonate 3.33% and Losartan 8.33%) was approximately 164%, relative to the immediate-release product (Cozaar). MRT and t(max) values were greater and C(max) values were lower for the GRDDS tablets compared with Cozaa. The lower bioavailability of Losartan in the CYP2C9*1/*1 subjects than CYP2C9*1/*3 subjects was found and could be due to the variety of enzymatic activity., (Copyright 2009 Elsevier B.V. All rights reserved.)

The thyroid hormone 3,3',5-triiodo-L-thyronine (T(3)) regulates growth, development, and differentiation processes in animals. These activities are mediated by the nuclear thyroid hormone receptors (TRs). Microarray analyses were performed previously to study the mechanism of regulation triggered by T(3) treatment in hepatoma cell lines. The results showed that spondin 2 was regulated positively by T(3). However, the underlying mechanism and the physiological role of T(3) in the regulation of spondin 2 are not clear. To verify the microarray results, spondin 2 was further investigated using semi-quantitative reverse transcription-PCR and western blotting. After 48 h of T(3) treatment in the HepG2-TR alpha 1#1 cell line, spondin 2 mRNA and protein levels increased by 3.9- to 5.7-fold. Similar results were observed in thyroidectomized rats. To localize the regulatory region in spondin 2, we performed serial deletions of the promoter and chromatin immunoprecipitation assays. The T(3) response element on the spondin 2 promoter was localized in the -1104/-1034 or -984/-925 regions. To explore the effect of spondin 2 on cellular function, spondin 2 knockdown cell lines were established from Huh7 cells. Knockdown cells had higher migration ability and invasiveness compared with control cells. Conversely, spondin 2 overexpression in J7 cells led to lower migration ability and invasiveness compared with control cells. Furthermore, this study demonstrated that spondin 2 overexpression in some types of hepatocellular carcinomas is TR dependent. Together, these experimental findings suggest that spondin 2, which is regulated by T(3), has an important role in cell invasion, cell migration, and tumor progression.

Background: Although performance measurement for assessing care quality is an emerging area, a system for measuring the quality of cancer care at the hospital level has not been well developed. The purpose of this study was to develop organization-based core measures for colorectal cancer patient care and apply these measures to compare hospital performance., Methods: The development of core measures for colorectal cancer has undergone three stages including a modified Delphi method. The study sample originated from 2004 data in the Taiwan Cancer Database, a national cancer data registry. Eighteen hospitals and 5585 newly diagnosed colorectal cancer patients were enrolled in this study. We used indicator-based and case-based approaches to examine adherences simultaneously., Results: The final core measure set included seventeen indicators (1 pre-treatment, 11 treatment-related and 5 monitoring-related). There were data available for ten indicators. Indicator-based adherence possesses more meaningful application than case-based adherence for hospital comparisons. Mean adherence was 85.8% (79.8% to 91%) for indicator-based and 82.8% (77.6% to 88.9%) for case-based approaches. Hospitals performed well (>90%) for five out of eleven indicators. Still, the performance across hospitals varied for many indicators. The best and poorest system performance was reflected in indicators T5-negative surgical margin (99.3%, 97.2%-100.0%) and T7-lymph nodes harvest more than twelve(62.7%, 27.6%-92.2%), both of which related to surgical specimens., Conclusions: In this nationwide study, quality of colorectal cancer care still shows room for improvement. These preliminary results indicate that core measures for cancer can be developed systematically and applied for internal quality improvement.

The simultaneous nitrogen and phosphorus removal (SNPR) from real domestic wastewater using aerobic granular sludge (AGS) has been investigated. The effect of temperature changes on the SNPR of AGS was also studied. Two lab-scale sequencing batch reactors (SBRs), named as A and B, were operated for 254 days in alternating anaerobic-aerobic mode. The AGS was cultivated in 20 days using settling time as selection pressure. From the 42nd day onwards, additional carbon (acetate and peptone for A, glucose for B) were added to make the ratio of COD:N:P as 360:60:6. The SNPR by AGS was achieved for 4 months, and the removal ratios of ammonia, total inorganic nitrogen and phosphorus of A were reached to 98.42%, 74.25% and 94.79%, respectively; the removal ratios of ammonia, total inorganic nitrogen and phosphorus of B were reached to 99.45%, 75.96% and 95.60%, respectively. Then, the temperature decreased to 9-13 degrees C, which seriously affected the efficiency of SNPR. The removal efficiencies of SNPR of reactor A and B were recovered after 35 days and 49 days, respectively, and the removal ratios of ammonia, total inorganic nitrogen and phosphorus of A were reached to 96.33%, 79.49% and 99.68%, respectively; the removal ratios of ammonia, total inorganic nitrogen and phosphorus of B were reached to 93.85%, 76.44% and 98.44%, respectively. PAOs and DNPAOs together completed the phosphorus removal and made phosphorus removal faster than nitrogen removal. Nitrification was the rate limiting step for SNPR. The low temperature made the anoxic zone in AGS decrease, together with the absence of PHAs, made denitrification could not complete.

The objective of this study was to identify genes regulated by thyroid hormone (T(3)) mediated by its receptor (TR) and associated with tumorigenesis. The gene encoding aldo-keto reductase family 1, member B1 (AKR1B1), as previously identified by c-DNA microarray, is known to be up-regulated by T(3) treatment. Enzyme AKR1B1 was elevated roughly 3-fold in HepG2-TRalpha1 cells at the protein level and 4.6-fold increase at the mRNA level after 48 h T(3) treatment. Similar findings were obtained from thyroidectomized rats after T(3) application. To identify and localize the critical TR element (TRE), series deletion of the promoter mutant were constructed and electrophoretic mobility shift assays were carried out. The TRE on the AKR1B1 promoter was localized to the -1099/-1028 region. Further, this study demonstrated that AKR1B1 over-expression in some types of hepatocellular carcinomas (HCCs) is TR-dependent and might play a crucial role in the development of HCC. Thus, T(3) regulates AKR1B1 gene expression via a TRE-dependant mechanism and associates liver cancer.

Objective: To observe therapeutic effect and analgesic eJfect of needling at Ashi point for treatment of piriformis syndrome., Methods: Eighty cases were randomly divided into an inhibitory-needling method group (INM) and a routine-needling method group (RNM), forty cases in each group. The acupuncture at Ashi point as major point with inhibitory-needling munipulation was applied in the INM group, the acupuncture at Huantiao (GB 30) ,Zhibian (BL 54) ,Yanglingquan (GB 34) with routine-needling method in the RNM group. The therapeutic effect, the instant analgesic effect and score of Visual Analogue Scale (VAS) were observed., Results: After treatment, the effective rate of 92.5% in the INM group was better than that of 82.5% in the RNM group (P < 0.05); the instant analgesic effect rate of 75.0% in the INM group was better than that of 52.5% in the RNM group (P < 0.01); the score of VAS in both groups had decreased with a significant difference between two groups (P < 0.05)., Conclusion: The treatment of inserting the needle at Ashi point as major point with inhibitory-needling method has excellent effectiveness and good instant analgesic effect, with fewer points selection and rapid effect.

The objective of this study was to identify genes regulated by thyroid hormone (T(3)) and associated with tumor invasion. The gene encoding furin, as previously identified by cDNA microarray, is known to be up-regulated by T(3) treatment, and stimulated furin production occurs in thyroidectomized rats after administration of T(3). Presently, by using serial deletion of the promoter and EMSAs, the T(3) response element on the furin promoter was localized to the -6317/-6302 region. T(3)-mediated furin up-regulation was cooperative with TGF-beta because T(3) induction increased after Smad3/4 addition. Furthermore, the invasiveness of HepG2-thyroid hormone receptor (TR) cells was significantly increased by T(3) treatment, perhaps due to furin processing of matrix metalloproteinase-2 and -9. In addition, furin up-regulation either by stable overexpression or T(3) and/or TGF-beta induction was evident in severe-combined immune-deficient mice inoculated with HepG2-TRalpha1 cells. The HepG2-furin mice displayed a higher metastasis index and tumor size than HepG2-neo mice. Notably, the increased liver and lung tumor number or size in the hyperthyroid severe-combined immune-deficient mice as well as TGF-beta mice was attributed specifically to furin overexpression in the HepG2-TRalpha1 cells. Furthermore, this study demonstrated that furin overexpression in some types of hepatocellular carcinomas is TR dependent and might play a crucial role in the development of hepatocellular carcinoma. Thus, T(3) regulates furin gene expression via a novel mechanism or in cooperation with TGF-beta to enhance tumor metastasis in vitro and in vivo.

Pituitary tumor transforming gene 1 (PTTG1) is expressed in most tumors. However, whether thyroid hormone (T(3)) and its receptors (TR) regulate PTTG1 in human hepatocellular carcinomas (HCC) remains unclear. Previous cDNA microarrays revealed PTTG1 is down-regulated by T(3)/TR. This study investigated the significance of PTTG1 regulation by T(3) in HCC cells. The PTTG1 mRNA and protein expression were repressed by T(3) in HCC cell lines overexpressing TR. However, after knockdown of TRs expression by RNA interference, PTTG1 repression by T(3) was abolished. Similar results were observed in thyroidectomized rats. To localize the regulatory region in the PTTG1 promoter, serial deletions within the PTTG1 promoter region were constructed. The promoter activity of the PTTG1 gene was repressed (25-51%) by T(3). Additionally, these findings indicate that PTTG1 may be regulated by Sp1. The critical role of the -594 and -520 Sp1 binding sites was confirmed by electrophoretic mobility shift assay. Transfection with Sp1 expression vector enhanced the activity of the PTTG1 promoter fragment reporter. Also, Sp1 was down-regulated in HCC cells and in thyroidectomized rat after T(3) treatment. Additionally, ectopic expression of PTTG1 promotes cell proliferation in Hep3B hepatoma cells. Conversely, knockdown of PTTG1 or Sp1 expression reduced cell proliferation in HepG2 cells. Notably, the expression of PTTG1 and Sp1 was inversely correlated with the expression of TR proteins in HCC. Together, these findings indicate that PTTG1 gene expression is mediated by Sp1 and is indirectly down-regulated by T(3). Finally, overexpression of PTTG1 or SP1 in HCCs is TR-dependent and crucial in the development of HCC.

Thyroid hormone (T3) regulates multiple physiological processes during development, growth, differentiation, and metabolism. Most T3 actions are mediated via thyroid hormone receptors (TRs) that are members of the nuclear hormone receptor superfamily of ligand-dependent transcription factors. The effects of T3 treatment on target gene regulation was previously examined in TRalpha1-overexpressing hepatoma cell lines (HepG2-TRalpha1). Androgen receptor (AR)-associated protein 70 (ARA70) was one gene found to be up-regulated by T3. The ARA70 is a ligand-dependent coactivator for the AR and was significantly increased by 4- to 5-fold after T3 treatment by Northern blot analyses in the HepG2-TRalpha1 stable cell line. T3 induced a 1- to 2-fold increase in the HepG2-TRbeta1 stable cell line. Both stable cell lines attained the highest fold expression after 24 h treatment with 10 nM T3. The ARA70 protein was increased up to 1.9-fold after T3 treatment in HepG2-TRalpha1 cells. Similar findings were obtained in thyroidectomized rats after T3 application. Cycloheximide treatment did not suppress induction of ARA70 transcription by T3, suggesting that this regulation is direct. A series of deletion mutants of ARA70 promoter fragments in pGL2 plasmid were generated to localize the thyroid hormone response element (TRE). The DNA fragments (-234/-190 or +56/+119) gave 1.55- or 2-fold enhanced promoter activity by T3. Thus, two TRE sites exist in the upstream-regulatory region of ARA70. The TR-TRE interaction was further confirmed with EMSAs. Additionally, ARA70 could interfere with TR/TRE complex formation. Therefore, the data indicated that ARA70 suppresses T3 signaling in a TRE-dependent manner. These experimental results suggest that T3 directly up-regulates ARA70 gene expression. Subsequently, ARA70 negatively regulates T3 signaling.

Two common symptoms of pregnancy are nausea and vomiting, which, for some women, cause great discomfort and profoundly impact daily life. A descriptive phenomenological method was used to understand how Taiwanese women deal with nausea and vomiting during pregnancy. A purposive sample of 10 expectant mothers participated in the study. A transcription of each informant's verbal description of her nausea and vomiting experience was made and analyzed using Colaizzi's phenomenological methodology. Four themes emerged from the interview data after inductively analyzing the narrative content concerning nausea and vomiting: 1) understanding nausea and vomiting, 2) finding coping strategies, 3) psychosocial adaptation, and 4) needing support. The results also indicated the importance of social support to pregnant women's health, including instrumental and emotional forms of support. Health professionals, however, were not mentioned in the mothers' narratives as a source of support. The issues of social support, language translation, and western and eastern perspectives on nausea and vomiting and cultural differences in perceptions of their severity are discussed.

Thyroid hormone, T(3), regulates cell metabolism, differentiation, and development. cDNA microarrays were performed to study the mechanism of target gene regulation after T(3) treatment in a thyroid hormone receptor-alpha (TRalpha)-overexpressing hepatoma cell line (HepG2-TRalpha). The differentially expressed target genes are several metabolic enzymes, including dehydroepiandrosterone-sulfotransferase family 1A member 2 (SULT2A1). Enzyme SULT2A1 was elevated roughly 5-fold at the protein level and 9-fold increase at the mRNA level after 48 h T(3) treatment in HepG2-TRalpha cells. Cycloheximide inhibited T(3)-induced SULT2A1 expression, suggesting that regulation was indirect. SULT2A1 has been reported to be regulated by the two transcription factors, steroidogenic factor 1 (SF1) and GATA, in the human adrenal gland. T(3) induced a 2.5- to 3.5-fold elevation of SF1 at the protein level and a 6.2-fold increase at the RNA level in HepG2-TRalpha cells. About seven SF1 binding sites exist on the SULT2A1 gene. To identify and localize the critical SF1 binding site, series of deletion mutants of SULT2A1 promoter fragments in pGL2 plasmid were constructed. The promoter activity of the SULT2A1 gene was enhanced about 2.8- to 7.1-fold by T(3). The -228 SF1 binding site was identified as the most critical site because deleting this region reduced T(3)-induced expression. Transcription factor SF1 application enhanced the -228 but not -117 reporter plasmid activities. SULT2A1 and SF1 up-regulation at protein and RNA levels in thyroidectomized rats occurred after T(3) application. In summary, this work demonstrated that the SULT2A1 gene was mediated by SF1 and indirectly regulated by T(3). Further study is required to elucidate the physiological importance of SULT2A1 induction mediated by T(3).

In an attempt to accelerate wound healing by stimulating the recruitment of fibroblasts and improve the mechanical properties of collagen matrixes, N,O-(carboxymethyl)chitosan (NOCC) was incorporated into the backbone of a collagen (COL) matrix without or with chondroitin sulfate (CS) or an acellular dermal matrix (ADM). The result of a cell migration study demonstrated that the migration of fibroblasts was significantly enhanced by NOCC in a concentration-dependent manner. In the analysis with a dynamic mechanical analyzer, NOCC/CS/COL matrixes presented higher tensile strengths than did NOCC/ADM/COL matrixes. Skin fibroblasts cultured on the matrixes containing NOCC showed increased proliferation and secretion of three kinds of cytokines compared with the control. Results of the in vivo wound healing study showed that matrixes incorporating NOCC showed markedly enhanced wound healing compared with the control. Therefore, the above results clearly suggest that NOCC/COL matrixes containing CS or ADM can be potential wound dressings for clinical applications.

In search of a new approach for crosslinking collagen-based biomaterials, we examined the effect of microbial transglutaminase (MTGases) as a crosslinking reagent on collagenous matrices made from porcine type I collagen. As the results revealed, MTGase exhibited a crosslinking action that raised the viscosity of the collagen solution. Matrices crosslinked with MTGase at the low pH values of pH 3 and 4 exhibited higher tensile strengths than those at high pH values. In comparison with untreated matrices, the denaturation temperatures of the corresponding matrices shifted toward higher temperatures. These enzyme-catalyzed crosslinked matrices were proven by MTT assay to be non-cytotoxic. In conclusion, this enzymatic method of using MTGase provides an alternative potential way for crosslinking collagen-based matrices.

Viscoelastic characteristics of acellular dermal matrix (ADM) preparations with various additives were analyzed with creep curves, stress-strain relationships, and the storage modulus with reference to those of ADM preparations crosslinked with glutaraldehyde. Creep curves for all ADM preparations were determined to comply with the Kelvin-Voigt model. The stress-strain plots of all ADM preparations compared were described as linear. The storage modulus of all ADM preparations was maintained at a nearly constant level throughout the range of oscillating frequencies applied. ADM preparations crosslinked with glutaraldehyde showed that both Young's modulus (E) for the spring part and retardation time (tau) in the Kelvin-Voigt model, and hence viscosity (eta) for the liquid part, increased with an increasing concentration of glutaraldehyde. Higher Young's modulus and viscosity and a greater extent of the "solid" response of ADM preparations crosslinked with glutaraldehyde might have been responsible for the longer persistence that was demonstrated after implantation. The increase in ADM concentration and the addition of various additives to ADM preparations, including alpha-hydroxy acid (citric acid, lactic acid, and glycolic acid) and hyaluronic acid, resulted in similar effects on the viscoelastic characteristics of the ADM preparations, but they were less efficacious than those crosslinked with glutaraldehyde. Among them, increasing ADM concentration to >200 mg/mL and addition of glycolic acid at a concentration of >2% improved the viscoelastic characteristics of the resulting ADM preparations so that their level of persistence was closer to that of material crosslinked with glutaraldehyde. On the contrary, the influence on viscoelastic characteristics of adding PVP greatly differed from that of hyaluronic acid and was only apparent when adding concentrations of PVP of >10%. Similarly, viscoelastic characteristics of the ADM preparations examined were also so sensitive to temperature that the persistence of ADM preparations after implantation at body temperature would deteriorate., (Copyright 2004 Wiley Periodicals, Inc.)

The object of this study was to compare the extent of decellularization at each critical step of processing porcine skin to produce an acellular dermal matrix (ADM) for biomedical applications. The results demonstrated that the removal of epidermis using treatment with 0.25% trypsin for 18 h and 0.1% sodium dodecyl sulfate (SDS) for 12 h at room temperature was beneficial for the subsequent treatment to remove cells in the dermal structure. Lengthy incubation in 0.25% trypsin (12 h) and then 560 units/l Dispase (12 h) at 25 degrees C of small pieces of porcine skin from which the epidermis had been removed efficiently removed cells and cellular components from the skin. Histological examinations revealed that the epidermis, dermal fibroblasts, and epidermal appendages were completely removed by these treatments, and the basic dermal architecture of collagen bundles was that of a loose meshwork. Examinations by TEM showed that the characteristics of collagen fibers in the ADM were retained after complete removal of cells present under optimal conditions defined in this study. SDS-PAGE and size-exclusion HPLC revealed that collagen fibers in the ADM were mostly type I and showed two typical component peaks identified as oligomers and monomers, respectively.

Objectives: Laparoscopic renal cryoablation is a developmental minimally invasive nephron-sparing treatment alternative for highly select patients with small renal tumors. We present our evolving experience with this procedure., Methods: Thirty-two patients (34 tumors) with a mean tumor size of 2.3 cm on preoperative computed tomography underwent laparoscopic renal cryoablation. As dictated by the tumor location, cryoablation was performed by either the retroperitoneal (n = 22) or the transperitoneal (n = 10) laparoscopic approach using real-time ultrasound monitoring. A double freeze-thaw cycle was routinely performed., Results: The mean surgical time was 2.9 hours, cryoablation time 15.1 minutes, and blood loss 66.8 mL. For a mean intraoperative ultrasonographic tumor size of 2 cm, the mean cryolesion size was 3.2 cm. The hospital stay was less than 23 hours in 22 (69%) of 32 patients. Sequential magnetic resonance imaging scans demonstrated a gradual contraction in the mean diameter of the cryolesions. Of the 20 patients who underwent a 1-year follow-up magnetic resonance imaging scan, the cryoablated tumor was no longer visible in 5. Of note, 23 patients have now undergone a 3 to 6-month follow-up computed tomography-directed biopsy of the cryoablated tumor site; the biopsy was negative for cancer in all 23 patients. No evidence of local or port-site recurrence was found during a mean follow-up of 16.2 months., Conclusions: Critical long-term data regarding laparoscopic renal cryoablation, a developmental technique, are awaited. However, our initial experience is cautiously optimistic. Despite its significant potential for false-negative results, it is encouraging that the follow-up computed tomography-directed needle biopsies at 3 to 6 months were negative for cancer in 23 of 23 patients.

Laparoscopic cryoablation seems to be an effective treatment modality for small peripheral renal tumors. The technique is minimally invasive, has a rapid learning curve, results in minimal blood loss and morbidity, and, to date, has demonstrated precise reliable ablation of small renal neoplasms. Long-term follow-up is necessary to confirm the absence of local tumor recurrence or distant or port-site metastases.

Objectives: Laparoscopy may be complicated by neuromuscular injuries, both to the patient and to the surgeon. We used a survey to estimate the incidence of these injuries during urologic laparoscopic surgery, to assess risk factors for these injuries, and to determine preventive measures., Methods: A survey of neuromuscular injuries associated with laparoscopy submitted to 18 institutions in the United States was completed by 18 attending urologists from 15 institutions., Results: From among a total of 1651 procedures, there were 46 neuromuscular injuries in 45 patients (2.7%), including abdominal wall neuralgia (14), extremity sensory deficit (12), extremity motor deficit (8), clinical rhabdomyolysis (6), shoulder contusion (4), and back spasm (2). Neuromuscular injuries were twice as common with upper retroperitoneal as with pelvic laparoscopy (3. 1% versus 1.5%). Among patients with neuromuscular injuries, those with rhabdomyolysis were heavier (means 91 versus 80 kg) and underwent longer procedures (means 379 versus 300 minutes), and those with motor deficits were older (means 51 versus 42 years of age). Of the surgeons, 28% and 17% reported frequent neck and shoulder pain, respectively., Conclusions: Although not common, neuromuscular injuries during laparoscopy do contribute to morbidity. Abdominal wall neuralgias, injuries to peripheral nerves, and joint or back injuries likely occur no more frequently than during open surgery, but risk of rhabdomyolysis may be increased. Positioning in a partial rather than full flank position may reduce the incidence of some injuries. Measures to reduce neuromuscular strain on the surgeon during laparoscopy should be considered.

Objectives: To assess retrospectively whether laparoscopic retroperitoneal lymph node dissection (RPLND) in patients with clinical Stage I nonseminomatous germ cell testicular tumor (NSGCT) provides useful pathologic staging information on which subsequent management can be based. Approximately 30% of patients with clinical Stage I NSGCT will have pathologic Stage II disease., Methods: A retrospective review of 29 patients with clinical Stage I NSGCT who underwent transperitoneal laparoscopic RPLND by a single surgeon was performed. Selection criteria included the presence of embryonal carcinoma in the primary tumor or vascular invasion. A modified left (n = 18) or right (n = 11) template was used., Results: Positive retroperitoneal nodes were detected in 12 (41%) of 29 patients. Ten of these patients received immediate adjuvant platinum-based chemotherapy, and 2 patients refused chemotherapy. The nodes were negative in 1 7 (59%) of 29 patients; all but 2 patients (one with recurrence in the chest, the other with biochemical recurrence) have undergone observation. No evidence of disease recurrence has been found in the retroperitoneum of any patient (follow-up range 1 to 65 months). Prospectively, the dissection was limited if grossly positive nodes were encountered; therefore, the total number of nodes removed was significantly different if the nodes were positive or negative (14 +/- 2 and 25 +/- 3, respectively; P <0.004). Two patients required an open conversion because of hemorrhage. Complications included lymphocele (n = 1) and flank compartment syndrome (n = 1)., Conclusions: Laparoscopic RPLND is a feasible, minimally invasive surgical alternative to observation or open RPLND for Stage I NSGCT. Disease outcomes are favorable to date. Longer follow-up in a larger series is necessary to determine therapeutic efficacy.

Purpose: We determine the subjective and objective durability of laparoscopic versus open pyeloplasty., Materials and Methods: From August 1993 to April 1997, 42 patients underwent laparoscopic pyeloplasty (laparoscopy group) with a minimum clinical followup of 12 months (mean 22). Subjective outcomes and objective findings were compared to those of 35 patients who underwent open pyeloplasty (open surgery group) from August 1986 to April 1997 with a minimum clinical followup of 12 months (mean 58). We assessed clinical outcome based on responses to a subjective analog pain and activity scale. In addition, radiographic outcome was assessed based on the results of the most recent radiographic study., Results: Of the 42 laparoscopy group patients 90% (38) were pain-free (26, 62%) or had significant improvement in flank pain (12, 29%) after surgery. Two patients had only minor improvement and 2 had no improvement in pain. Surgery failed in only 1 patient with complete obstruction. A patent ureteropelvic junction was demonstrated in 98% (41 of 42 patients) of the laparoscopy group on the most recent radiographic study (mean radiographic followup 15 months). Of the 35 open surgery group patients 91% were pain-free (21, 60%) or significantly improved (11, 31%) after surgery. One patient had only minor improvement and 2 were worse., Conclusions: Pain relief, improved activity level and relief of obstruction outcomes are equivalent for laparoscopic and open pyeloplasty.

Background and Objectives: The aim of minimally invasive approaches to vesicoureteral reflux, such as endoscopic trigonoplasty, is to lower the morbidity of open procedures without compromising the results. Initial successes have not been sustained, mainly because of trigonal splitting, which results in the ureteral orifices returning to their preoperative positions. This study was designed to address trigonal splitting by mobilizing the ureters before repositioning them and to evaluate the feasibility of accomplishing this intravesically with 2- to 3-mm endoscopic mini-instruments., Methods: Bilateral vesicoureteral reflux was surgically created in 10 minipigs. After radiologic confirmation of success 4 weeks later, modified trigonoplasty was performed by endoscopic intravesical mobilization of both ureters and incision of the trigonal mucosa using 2-mm instruments. The ureteral orifices were then advanced toward the midline and sutured in place. The initial surgical techniques were modified to permit the entire procedure to be performed endoscopically in the last four minipigs. Cystograms and intravenous urograms were obtained 4 weeks later., Results: Two minipigs died postoperatively. Six of the remaining eight had persistent reflux, including three of the four in the group treated completely by endoscopic means. None of the dissected ureters showed evidence of stricture or necrosis., Conclusions: Although the procedure was not successful in correcting reflux in this model, this study demonstrates the feasibility of endoscopic ureteral mobilization. With current instrumentation, there is no significant technical obstacle to complete intravesical endoscopic surgery, including ureteral reimplantation.

Laparoscopic partial nephrectomy (LPN) remains a technically challenging procedure largely because of the lack of methods for obtaining consistent parenchymal hemostasis. The objective of this study was to determine if the extent of resection influences the ability of the harmonic scalpel to achieve hemostasis and to define the cases in which the harmonic scalpel is appropriate for LPN. Thirty LPNs were performed in a 25-kg domestic pig model. The blunt blade of the laparoscopic harmonic scalpel (LaparoSonic Coagulating Shears; Ethicon Endo-Surgery, Cincinnati, OH) at power level 5 was used to divide the parenchyma. Control of the renal hilar vessels was not obtained. Three standardized types of resections were performed: I = peripheral wedge biopsy; II = upper- or lower-pole nephrectomy; and III = heminephrectomy. Bleeding was graded on a scale from 0 to 4: 0 = no hemostasis; 1 = steady bleeding; 2 = moderate bleeding; 3 = parenchymal oozing; and 4 = dry. Hemostasis grades of 2 or less were clinically significant bleeding necessitating supplemental coagulation. The mean hemostasis scores showed a significant (P < 0.02) trend toward inadequate hemostasis with increasing extent of resection: 3.3 for Type I, 3.0 for Type II, and 2.4 for Type III. The percent of kidneys with grade 2 bleeding or worse was 9% for Type I surgery, 25% for Type II, and 57% for Type III. Successful hemostasis with the harmonic scalpel correlates with the extent of parenchymal resection in the porcine model. Most wedge excisions can be done with the harmonic scalpel alone, whereas larger resections necessitate supplemental coagulation. On the basis of this study, heminephrectomies with the harmonic scalpel are not recommended because of the high incidence of significant hemorrhage.

Background: Renal biopsy continues to be a pivotal tool and frequently indispensable diagnostic procedure in the clinical assessment of proteinuria and or unexplained renal disease. Laparoscopic renal biopsy has recently been reported as an alternative to open renal biopsy., Methods: Thirty-two patients who had proteinuria and/or renal insufficiency underwent laparoscopic renal biopsy at our center. The indications for biopsy included failed percutaneous biopsy (N = 3), morbid obesity (14), solitary kidney (5), chronic anticoagulation/coagulopathy (6), religious consideration (refusal of potential blood transfusion) (2), multiple bilateral renal cysts and body habitus (1 case each). The kidney was approached via a laparoscopic retroperitoneal route (retroperitoneoscopy) using a two port technique. The lower pole of the kidney was localized using blunt dissection, laparoscopic cup biopsies were performed, and hemostasis was achieved using standard techniques., Results: All biopsies were successfully completed laparoscopically with sufficient tissue obtained for histopathological diagnosis in all cases. Mean estimated blood loss was 25.9 ml (range 5 to 100). None of the patients required parenteral narcotics during the perioperative period. Operative time ranged from 0.8 to 3.0 hours (mean 1.5). Mean hospital stay was 1.7 days (range 0 to 7). Sixteen patients were treated as outpatients. Patients returned to normal activity at a mean of 1.7 weeks (range 0.3 to 3.0) postoperatively. In one patient, the spleen was inadvertently biopsied without consequence. An additional patient developed a postoperative 300 cc perinephric hematoma that resolved without the need for intervention. One postoperative mortality occurred on postoperative day seven secondary to a perforated peptic ulcer in a patient undergoing high-dose steroid therapy for lupus nephritis., Conclusion: Laparoscopic renal biopsy is a safe, reliable, minimally invasive alternative to open renal biopsy for patients in whom a closed percutaneous approach is either a relative or absolute contraindication, which can be performed on an outpatient basis.

A 26-year-old patient with end-stage renal disease secondary to vesicoureteral reflux and recurrent pyelonephritis was referred for bilateral native nephrectomy. A transperitoneal laparoscopic approach was used. Extremely dense fibrosis was encountered around the left kidney during the dissection. A left laparoscopic subcapsular nephrectomy and a right extracapsular nephrectomy were performed. The indications and surgical technique for laparoscopic subcapsular nephrectomy are discussed.

Laparoscopic pyeloplasty is one of several minimally invasive treatment options for UPJ obstruction. In fact, several endoscopically and fluoroscopically controlled methods of incising the obstructed UPJ are now available that are significantly less invasive and less morbid in comparison with open pyeloplasty. However, the long-term success rates of these incisional techniques are less than the rates reported for open pyeloplasty. Several causes of obstruction may be present in the primarily obstructed UPJ, including kinking or compression related to crossing vessels or intrinsic narrowing at the UPJ. One potential reason for the inferior success rates of incisional methods in comparison with open pyeloplasty is that the former techniques address the intrinsically narrowed UPJ but may not address extrinsic problems such as kinking of the ureter associated with fibrotic bands or compression from crossing vessels. Laparoscopic pyeloplasty addresses all potential causes of obstruction. Any fibrotic bands kinking the ureter are divided, and the ureter is spatulated through the level of the UPJ prior to completion of the anastomosis. If a crossing vessel is encountered, a dismembered pyeloplasty is performed, the ureter and renal pelvis are transposed to the opposite side of the vessels, and the anastomosis is completed. An additional disadvantage of incisional techniques is the significant risk of hemorrhage following incision of the UPJ, with as many as 3% to 11% of patients requiring blood transfusion. Hemorrhage may occur owing to an errant anterior incision, the presence of a crossing vessel, incision into the renal parenchyma adjacent to the UPJ, or as the result of bleeding from the percutaneous access site. In contrast, mean estimated blood loss in the authors' series of 57 laparoscopic pyeloplasties was 139 mL, and none of the patients required blood transfusion. Although it is more morbid in comparison with retrograde or fluoroscopically controlled endopyelotomy, laparoscopic pyeloplasty seems at least comparable to antegrade percutaneous endopyelotomy in terms of the length of hospitalization and patient convalescence. Laparoscopic pyeloplasty, however, offers a higher success rate than with incisional techniques, not only from a radiographic standpoint but from a subjective standpoint as determined by the results of the analogue pain and activity questionnaire. The major disadvantage of laparoscopic pyeloplasty is the need for proficiency in laparoscopic techniques and for a longer operative time. As a result, the literature on laparoscopic pyeloplasty consists primarily of small series. Janetschek and co-workers reported on a series of 17 patients who underwent laparoscopic pyeloplasty, including 14 via a transperitoneal approach and 3 via a retroperitoneal approach. Procedures performed included ureterolysis alone, dismembered pyeloplasty, and nondismembered (Fenger) pyeloplasty. "Fenger-plasty" is similar to Y-V pyeloplasty and is performed by incising the UPJ longitudinally and closing the incision transversely in a Heineke-Mikulicz fashion. Janetschek and colleagues reported a 100% success in the eight patients who underwent dismembered pyeloplasty but believed that this technique was too cumbersome and should be reserved for patients with long stenoses, dorsally crossing vessels, or large renal pelvis. Because two of the four patients undergoing ureterolysis alone failed treatment, Janetschek and colleagues have abandoned this technique. They now prefer the Fenger-plasty technique, even in the setting of ventrally crossing vessels, because the technique can be performed quickly with one to three sutures, and the anastomosis can be sealed with fibrin glue and a flap of Gerota's fascia. Their experience with this technique, however, remains relatively limited. Technologic advances such as the Endostitch device have facilitated reconstructive laparoscopic procedures such as pyeloplasty. (ABSTRACT TRUNCATED)

Obtaining accurate percutaneous renal access when treating intrarenal disease requires substantial skill. Robotic devices have been used in a variety of surgical applications and have been successful in facilitating percutaneous puncture while improving accuracy. Laboratory models of robotic devices for percutaneous renal access have also been developed. However, several technical hurdles need to be addressed. One relates to the device-patient interface. As a first step in creating a complete robotic system, a mechanical arm (PAKY) with active translational motion for percutaneous renal access has been developed and clinically assessed. The PAKY consists of a passive mechanical arm mounted on the operating table and a radiolucent needle driver that uses a novel active translational mechanism for needle advancement. The system utilizes real-time fluoroscopic images provided by a C-arm to align and monitor active needle placement. In vitro experiments to test needle placement accuracy were conducted using a porcine kidney suspended in agarose gel. Seven copper balls 3 to 12.5 mm diameter were placed in the collecting system as targets, and successful access was confirmed by electrical contact with the ball. The PAKY was then used clinically in nine patients. The number of attempts, target calix location, calix size, and time elapsed were evaluated. In the in vitro study, successful needle-ball contact occurred the first time in all 70 attempts, including 10 attempts at the 3-mm balls. Clinically, percutaneous access to the desired calix was attained on the first attempt in each case. The mean target calix diameter was 14.7 mm (range 7-40 mm). The mean time elapsed while attempting access was 8.2 minutes. No perioperative complications attributable to needle access occurred. Early experience indicates that the PAKY provides a steady needle holder and an effective and safe end-effector for percutaneous renal access. This device may provide the mechanical platform for the development of a complete robotic system capable of creating percutaneous renal access.