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Acute myocardial infarction (AMI) is a major cause of death worldwide. In the USA, there are approximately 0.8 million persons suffering from AMI annually with a death rate of 27%. The risk factors of AMI include hypertension, family history, smoking habits, diabetes, serenity, obesity, cholesterol, alcoholism, coronary artery disease, and so forth. In this study, data acquired from a subset of the National Health Insurance Research Database (NHIRD) of Taiwan were used to develop a clinical decision support system (CDSS) to predict AMI. The integrated genetic algorithm and support vector machine (IGS) and deep neural network (DNN) were both applied to design the predictive models. A balanced dataset (6087 AMI patients and 6087 non-AMI patients) and an imbalanced dataset (6,087 AMI patients and 12,174 non-AMI patients) with each patient record including 74 features were retrieved to design the predictive models. Tenfold cross-validation was used to obtain the optimal model with best prediction performance during training. The experimental results showed that the CDSSs reached a prediction performance with accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve (AUC) of 79.75–84.4%, 68.29–83.7%, 82.45–92.07%, and 0.8424–0.9089, respectively, for models designed based on the balanced dataset, as well as 81.86–86.27%, 52.65–81.22%, 84.29–96.47%, and 0.8503–0.9098, respectively, for models implemented based on the imbalanced dataset. The IGS and DNN algorithms and a combination of age, presence of related comorbidities, and other comorbidity-related features, including diagnosed age and annual physician visits of individual comorbidities, have been shown to be promising in designing strong predictive models in detecting patients who are more likely to develop AMI in the near future as well as for realizing mobile-health (m-Health) systems in managing their comorbidities to prevent occurrence of AMI events. Future work will focus on realizing an ensemble model by combining the model designed based on the long-term NHIRD dataset and the model based on the short-term EMR data and the real-time IoT sensor data, as well as implementing a transfer learning model by transferring the knowledge learned from the long-term model for training the short-term model, so that the predictive performance can be enhanced.

Background Percutaneous endoscopic surgery is a popular surgery to treat lumbar spinal disorders. However, seizure after percutaneous endoscopic surgery is an unpredictable complication. The only prodromal sign for seizure currently known is neck pain. We reviewed the incidence of, and risk factors for, seizure during percutaneous endoscopic surgery and present the cases of 3 patients with seizure and our management. Case Description From October 2006 to March 2019, 3 of 816 patients (0.34%) with thoracic lumbar disorders who had undergone percutaneous endoscopic surgery experienced a seizure episode. The cases of those 3 patients were carefully reviewed. Studies of the risk factors for seizure after spinal procedures reported before June 13, 2019 were identified through a PubMed search. We found that infusion fluid containing cefazolin, the infusion rate, a prolonged operative time, the occurrence of a dural tear, and sevoflurane anesthesia might be associated with seizure, both described in the reported data and found in our experience. Three patients who experienced a seizure episode had had general anesthesia with sevoflurane, and the surgical approach used was interlaminar for a herniated disc in L5-S1. We noted a “red flag sign,” namely an uncontrollable hypertension episode combined with a decreasing pulse rate, in all 3 patients who had experienced a seizure, which was not observed in the other patients. All 3 patients had received antihypertensive medication (labetalol) ≥3 times without response. Conclusion Seizure after percutaneous endoscopic surgery is rare, but lethal. Although its cause remains unknown, all risk factors for seizure should be checked and corrected immediately when a red flag sign, uncontrolled hypertension, appears.

Background Surgical interventions for congenital scoliosis are challenging for spine surgeons. The coordination of 3-dimensional (3D), real-time, image-guided navigation with an ultrasonic bone cutter allows surgeons to localize the affected area of the spine accurately and remove the lesion without damaging soft tissue structures. The goal of this technical paper is to report a previously undescribed method of hemivertebrectomy that combines 3D, real-time, image-guided navigation and an ultrasonic bone cutter. We highlight the feasibility and safety of this method in spinal surgery. Methods Three patients with congenital scoliosis were treated with this technique. We present three illustrative cases comprising hemivertebrectomies for congenital scoliosis. Intraoperative photos demonstrating the technique are also provided. Results All surgeries were completed without complications. The hemivertebrae were completely removed, and marked correction of congenital scoliosis was noted. Conclusions We believe that the combination of 3D, real-time image navigation and an ultrasonic bone cutter improves hemivertebrectomy by increasing accuracy and avoiding dura laceration, major organ damage, or potential vessel damage.

Background The risk of injury directly related to hospitalization for motor vehicle accidents (MVAs) in the obstructive sleep apnea (OSA) patients has not been thoroughly understood. Our study aimed to examine the association between the OSA and the hospitalization for an MVA injury. Methods This retrospective cohort study used Taiwan’s National Health Insurance Research Database (NHIRD) between 2000 and 2015. The OSA patients aged ≥20 years by age, sex, and index-year matched by non-OSA controls were enrolled (1:3). We used the Cox proportional regression model to evaluate the association between the OSA and the hospitalization for an MVA injury. Results The incidence rate of hospitalization for an MVA injury was higher in the OSA cohort (N = 3025) when compared with the non-OSA controls (N = 9075), as 575.3 and 372.0 per 100,000 person-years, respectively (p p p Conclusions This study provides evidence that patients with OSA are at a two-fold higher risk of developing hospitalization for an MVA injury, and the usage of modafinil and methylphenidate was associated with a lower risk of an overall hospitalization for the MVA injury.

This study processed the recent in vivo survey results for over a thousand patients and optimized their neck and head CT angiography triggered timing (CTA-TT) via the inverse problem algorithm, which ensured the maximal ratio of both left and right arterial to upper sinuses (LRA/US). These results are instrumental in examining the ischemic stroke syndromes along the neck and head. These 1001 patients were randomly categorized into test surveyed (802 patients) and verification group (199 patients), then a six factors semi-empirical formula was constructed by the STATISTICA program. The six factors were assigned a patient’s biological data and preset of the CTA facility; namely Age, mean arterial pressure (MAP), heart rate (HR), contrast media dose (CMD), Pre (injected pressure of CMD), and body surface area (BSA). Each factor was normalized into dimensionless values and incorporated into the dataset matrix [Formula: see text] to analyze the coefficient matrix [Formula: see text]. The derived semi-empirical formula closely correlated with experimental data, according to the loss function [Formula: see text], correlation coefficient [Formula: see text], and variance of 0.8965. The formula verification for 199 more patients (verification group) yielded a correlation coefficient [Formula: see text]. Thus, it can be used for the CTA-TT estimation of patients without their preliminary tests, avoiding unnecessary irradiation. The estimated LRA/US was [Formula: see text] for the verification group in this study. A simplified three-factor formula, featuring only age, MAP, and BSA, was also proposed.

The COVID-19 pandemic is an ongoing global pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in 2020–2021. COVID-19 is becoming one of the most fatal pandemics in history and brings a huge challenge to the global healthcare system. Opportune detection, confinement, and early treatment of infected cases present the first step in combating COVID-19. Diagnosis via viral nucleic acid amplification tests (NAATs) is frequently employed and considered the standard procedure. However, with an increasing urge for point-of-care tests, rapid and cheaper immunoassays are widely utilized, such as lateral flow immunoassay (LFIA), which can be used for rapid, early, and large-scale detection of SARS-CoV-2 infection. In this narrative review, the principle and technique of LFIA applied in COVID-19 antigen and antibody detection are introduced. The diagnostic sensitivity and specificity of the commercial LFIA tests are outlined and compared. Generally, LFIA antigen tests for SARS-CoV-2 are less sensitive than viral NAATs, the “gold standard” for clinical COVID-19 diagnosis. However, antigen tests can be used for rapid and mass testing in high-risk congregate housing to quickly identify people with COVID-19, implementing infection prevention and control measures, thus preventing transmission. LFIA anti-SARS-CoV-2 antibody tests, IgM and/or IgG, known as serology tests, are used for identification if a person has previously been exposed to the virus or vaccine immunization. Notably, advanced techniques, such as LFT-based CRISPR-Cas9 and surface-enhanced Raman spectroscopy (SERS), have added new dimensions to the COVID-19 diagnosis and are also discussed in this review.

Background. The pedicle screw is one of the main tools used in spinal deformity correction surgery. Robotic and navigated surgeries are usually used, and they provide superior accuracy in pedicle screw placement than free-hand and fluoroscopy-guided techniques. However, their high cost and space limitation are problematic. We provide a new solution using 3D printing technology to facilitate spinal deformity surgery. Methods. A workflow was developed to assist spinal deformity surgery using 3D printing technology. The trajectory and profile of pedicle screws were determined on the image system by the surgical team. The engineering team designed drill templates based on the bony surface anatomy and the trajectory of pedicle screws. Their effectiveness and safety were evaluated during a preoperative simulation surgery. The surgery consisted in making a pilot hole through the drill template on a computed tomography- (CT-) based, full-scale 3D spine model for every planned segment. Somatosensory evoke potential (SSEP) and motor evoke potential (MEP) were used for intraoperative neurophysiological monitoring. Postoperative CT was obtained 6 months after the correction surgery to confirm the screw accuracy. Results. From July 2015 to November 2016, we performed 10 spinal deformity surgeries with 3D printing technology assistance. In total, 173 pedicle screws were implanted using drill templates. No notable change in SSEP and MEP or neurologic deficit was noted. Based on postoperative CT scans, the acceptable rate was 97.1% (168/173). We recorded twelve pedicle screws with medial breach, six with lateral breach, and five with inferior breach. Medial breach (12/23) was the main type of penetration. Lateral breach occurred mostly in the concave side (5/6). Most penetrations occurred above the T8 level (69.6%, 16/23). Conclusion. 3D printing technology provides an effective alternative for spinal deformity surgery when expensive medical equipment, such as intraoperative navigation and robotic systems, is unavailable.

Ischemic stroke (IS) is the most common type of stroke, accounting to about 80% of the stroke, and is a major cause of morbidity and mortality worldwide. The risk factors of IS included older age, male gender, high BMI, smoking habit, hypertension, diabetes, hyperlipidemia, etc. Other factors, such as environmental metal exposure and atrial cardiopathy are also found to be risk factors. In this study, a balanced dataset, consisting of healthcare data of 36,880 IS patients and 36,880 non-IS patients matched with IS patients using indexed date, age, and sex, retrieved from a subset of the National Health Insurance Research Database (NHIRD) of Taiwan, an administrative healthcare database, were adopted for developing the AI models to predict events of ISs. Integrated Genetic Algorithm and Support Vector Machine (IGS) algorithm accompanied with 3 different fitness functions was applied for designing the predictive models. To select the best predictive performance from different AI models, tenfold cross validation were conducted for model training. The predictive performance of the designed models exhibits that predictive accuracy, sensitivity, specificity, and area under ROC curve (AUC) achieve 73.38-73.96%, 73.31-73.91%, 73.03-74.02%, and 0.808-0.813, respectively. The selected features including age, comorbidities, and other comorbidity-related variables are shown to be effective in designing strong predictive models (AUC>0.8) for predicting patients who are more likely to develop IS in the near future. Future works will focus on designing the predictive models using more effective AI technique, such as deep neural network (DNN), and useful variables, such as administrated drugs, to enhance the predictive performance.

Background The aim of this study is to introduce a new method of percutaneous endoscopic decompression under 3D real-time image-guided navigation for spinal stenosis in degenerative kyphoscoliosis patients without instability or those who could not endure major surgery due to multiple comorbidities. Decompression along using endoscope for kyphoscoliosis patient is technical demanding and may result in unnecessary bone destruction leading to further instability. The O-arm/StealthStation system, a 3D real-time image-guided navigation system, is popular for its ability to provide automated registration with intraoperative, postpositioning computed tomography (CT) with no radiation exposure to operative staffs. With 3D real-time image-guided navigation, we could improve the accuracy of surgery and maintain the spinal stability as preoperative condition. Methods In this study, we presented four cases. All patients were over seventy years old female with variable degrees of kyphoscoliosis and multiple comorbidity who could not endure major spine fusion surgery. Percutaneous endoscopic unilateral laminotomy and bilateral decompression under 3D real-time image-guided navigation were successfully performed with following steps. First, the reference pin was introduced over iliac crest. Second, all navigational instruments were registered to the O-arm Stealth Station. Third. With the interlaminar approach, a 8.0 mm dilator followed by operative sleeve was introduced toward the ligamentum flavum. Then the endoscope attached with universal tracker was inserted, and the boundaries of lamina and facet joint were well-identified through the monitor. Finally, unitaleral laminotomy and resection of ligamentum flavum with joy-stick technique for bilateral decompression were performed under the navigation system. Patients’ demographics, image study parameters, and outcome measurements including pre- and post-operative serial visual analog scale (VAS), and Oswestry Disability Index (ODI) were well documented. The follow-up time was one year. Results All patients were successfully treated with this technique. Pre- and post-operative MRI showed average dural sac cross sectional area (DSCSA) improved from 81.62 (range 67.34–89.07) to 153.27 (range 127.96-189.73). All preoperative neurological symptoms including radicular leg pain and sciatica improved postoperatively. The mean ODI (%) were 85 (range 82.5–90) at initial visit, 35.875 (range 25–51) at 1 month post-operatively, 26.875 (range 22.5–35) at 6 months post-operatively and 22.5 (range 17.5–30) at 12 months post-operatively (p p

Readmission of patients is generally caused by inappropriate inpatient care, unexpected disease deterioration, inadequate health care plans for transition to community care, or premature discharge, and may be manifested by a degradation of healthcare quality and an increase of healthcare costs. In this study, we aimed to design a clinical decision support system (CDSS) based on patient data collected in Taiwan National Health Insurance Research Database in the prediction of 30-day all-cause readmissions for patients admitted with all-cause conditions. In order to select significant variables and adjust appropriate model parameters, we applied a method integrating genetic algorithm and support vector machine to design CDSSs based on 3 different cost-sensitive objective functions. The imbalanced data of patients admitted with all-cause conditions, including 6864 cases of admission and 48,374 cases of non-admission in 2011 as well as 6923 admissions and 49,521 non-admissions in 2012 were retrieved and used for training and testing the CDSSs, respectively. Each patient data consist of 31 candidate variables, including age, gender, number of comorbidities, Charlson comorbidity index (CCI), presence of comorbidities for calculating CCI, events within 1 year before admission, inpatient intervention, category of admitted hospital, length of admission, and discharge status. In the training phase, cluster-based under-sampling method (kNN) was used to prepare the balanced dataset for training and validating the CDSS. In contrast, the imbalanced testing dataset was applied to test the predictive performance of the trained CDSS model. The accuracy, sensitivity, specificity, and area under ROC curve (AUC) of the CDSSs designed with tenfold cross validation are 69.0–69.4%, 69.3.9–69.7%, 68.8–69.4%, and 0.74–0.76, respectively, in the training phase, and 62.3–64.3%, 68.0–69.5%, 61.3–63.8%, and 0.70–0.72, respectively, in the testing phase. Compared with the model in Maali et al. (BMC Med Inform Decis Making 18(1):1, 2018) (training phase: 0.74; testing phase: 0.71), all three models exhibit higher sensitivity and greater AUC. Among the 31 variables, 10, 18, and 17 salient variables, respectively, were selected for designing the models with three different objective functions. An online web service system designed based on Model 2 is provided for assisting physicians to detect potential patients having higher risk of all-cause readmissions after discharge and, if necessary, give them appropriate interventions to reduce their morbidities and mortalities as well as to reduce their healthcare costs.

Acute myocardial infarction (AMI) is a major cause of death worldwide. There are around 0.8 million persons suffered from AMI annually in the US and the death rate reaches 27%. The risk factors of AMI were reported to include hypertension, family history, smoking habit, diabetes, serenity, obesity, cholesterol, alcoholism, coronary artery disease, etc. In this study, data acquired from a subset of the National Health Insurance Research Database (NHIRD) of Taiwan were used to develop the clinical decision support system (CDSS) for predicting AMI. Support vector machine integrated with genetic algorithm (IGS) was adopted to design the AMI prediction models. Data of 6087 AMI patients and 6087 non-AMI patients, each includes 50 features, were acquired for designing the predictive models. Tenfold cross validation and three objective functions were used for obtaining the optimal model with best prediction performance during training. The experimental results show that the CDSSs reach a prediction performance with accuracy, sensitivity, specificity, and area under ROC curve (AUC) of 81.47–84.11%, 75.46–80.94%, 86.48–88.21%, and 0.8602–0.8935, respectively. The IGS algorithm and comorbidity-related features are promising in designing strong CDSS models for predicting patients who may acquire AMI in the near future.

Objective To focus on the potential beneficial effects of the pleiotropic effects of dipeptidyl peptidase-4 inhibitors (DPP4is) on attenuating progression of diabetic kidney disease in reducing the long-term effect of the acute kidney injury (AKI) to chronic kidney disease (CKD) transition. Patients and Methods Data from the National Health Insurance Research Database from January 1, 1999, to July 31, 2011, were analyzed, and patients with diabetes weaning from dialysis-requiring AKI were identified. Cox proportional hazards models and inverse-weighted estimates of the probability of treatment were used to adjust for treatment selection bias. The outcomes were incident end-stage renal disease (ESRD) and mortality, major adverse cardiovascular events, and hospitalized heart failure. Results Of a total of 6165 patients with diabetes weaning from dialysis-requiring AKI identified, 5635 (91.4%) patients were DPP4i nonusers and 530 (8.6%) patients were DPP4i users. Compared with DPP4i nonusers, DPP4i users had a lower risk of ESRD (hazard ratio, 0.81; 95% CI, 0.70-0.94; P=.04) and all-cause mortality (hazard ratio, 0.28; 95% CI, 0.23-0.34; P

Gold nanoparticles (AuNPs) can be applied in biosensors using fluorescence resonance energy transfer (FRET) technique. Based on this technique, we have established a sensitive and efficient biosensing method by modifying a peptide-probe onto AuNPs to detect proteinase enzyme activity in this study. This biosensing method was designed for chymase activity detection and applied in kidney disease diagnosis. In this study, 16 nm-AuNPs were used to construct the AuNPs-based fluorescence peptide probe (named AuNPs-peptide probe) for chymase activity determination. The peptide sequence is FITC-Acp-DRVYIHPFHLDDDDDC, which comprises a fluorophore at the N-terminal end, an enzyme (chymase) substrate (DRVYIHPFHL), a spacer (DDDDD) and cysteine (C) to conjugate to AuNPs surface. When the enzyme catalyzes the substrate sequence, the fluorophore drifts away from AuNPs and the fluorescence emitting signal can be excited at 495 nm and detected at 515 nm. The results indicate that the time required for the AuNPs-peptide probe for activity detection of chymase was only 15 min, and a linear correlation from 10 to 100 ng mL−1 of chymase was acquired. The chymase reaction would be significantly inhibited by addition of specific chymase inhibitor chymostatin. The AuNPs-peptide probe was tested for the detection of high concentrations of trypsin and chymotrypsin, but only minor emitted fluorescence intensity was detected. According to these results, sensitivity and specificity of the AuNPs-peptide probe for chymase detection have been confirmed. AuNPs-peptide probe was successfully used for the detection of renal chymase activity; and the results indicate the pathogenically increased chymase activity in kidney tissue of nephropathic mice from aristolochic acid I treatment.

// Cheng-Yi Chen 1, 2, 3 , Cheng-Jui Lin 2, 4, 5 , Chih-Sheng Lin 3 , Fang-Ju Sun 2, 6 , Chi-Feng Pan 2, 4 , Han-Hsiang Chen 2, 4 and Chih-Jen Wu 4, 5, 7, 8 1 Division of Nephrology, Department of Internal Medicine, Mackay Memorial Hospital, Hsinchu, Taiwan 2 MacKay Junior College of Medicine, Nursing and Management, Taipei, Taiwan 3 Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan 4 Division of Nephrology, Department of Internal Medicine, Mackay Memorial Hospital, Taipei, Taiwan 5 Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan 6 Department of Medical Research, MacKay Memorial Hospital, Taipei, Taiwan 7 Graduate Institute of Medical Sciences and Department of Pharmacology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan 8 Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, Taiwan Correspondence to: Chih-Jen Wu, email: wcjyali@yahoo.com.tw Keywords: liver cirrhosis; chronic kidney disease; diabetes mellitus; estimated glomerular filtration rate; MELD score Received: June 02, 2017 Accepted: August 29, 2017 Published: December 18, 2017 ABSTRACT Background: Chronic kidney disease (CKD) in cirrhosis is one of the dreaded complications associated with a steep rise in mortality and morbidity, including diabetes. There are limited data on the prevalence of CKD and the association with diabetes in outpatients with cirrhosis. Methodology: This is a cross-sectional study of 7,440 adult liver cirrhosis patients enrolled from August 2001 to April 2010 in a medical center. Case control matching by age and sex with 1,967 pairs, and conditional logistic regression for odds of diabetes was analyzed using adjusted model. Results: CKD was present in 46.0%, 45.7% and 45.6% of the study population using the MDRD-6, CKD-EPI and MDRD-4 estimated glomerular filtration rate (eGFR) equations, respectively. Using a conditional logistic regression model after adjusting for other risk factors, odds for diabetes increased significantly compared with non-CKD in CKD stage 3 to 5 (stage 3~5) based on MDRD-6–adjusted model, ORs were: stage 3~5, 2.34 (95% CI, 1.78-3.01); MDRD-4–adjusted model, ORs were: stage 3~5, 8.51 (95% CI, 5.63-11.4); CKD-EPI–adjusted model, ORs were: stage 3~5, 8.61 (95% CI, 5.13-13.9). Conclusion: In cirrhosis patients, prevalence of diabetes was higher in patients with advanced stage of CKD. For patients with cirrhosis, patients with CKD stages 3~5 defined by MDRD-4, MDRD-6, and CKD-EPI eGFR equations had increased risk for diabetes. More severe cirrhosis, indicated by the Child-Turcott-Pugh classification was also accompanied by an increased risk for diabetes.

// Chih-Jen Wu 1, 2, 3 , Cheng-Yi Chen 1, 8, 11 , Thung-S. Lai 4 , Pei-Chen Wu 1 , Chih-Kuang Chuang 5, 6, 7 , Fang-Ju Sun 8, 9 , Hsuan-Liang Liu 5 , Han-Hsiang Chen 1, 8 , Hung-I. Yeh 10 , Chih-Sheng Lin 11 and Cheng-Jui Lin 1, 5, 8 1 Division of Nephrology, Department of Internal Medicine, Mackay Memorial Hospital, Mackay Medical College ,Taipei, Taiwan 2 Department of Medical Research, China Medical University Hospital, China Medical University, Taichung, Taiwan 3 Graduate Institute of Medical Sciences and Department of Pharmacology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan 4 Graduate Institute of Biomedical Science, Mackay Medical College, New Taipei City, Taiwan 5 Institute of Biotechnology, National Taipei University of Technology, Taipei, Taiwan 6 Division of Genetics and Metabolism, Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan 7 College of Medicine, Fu-Jen Catholic University, Taipei, Taiwan 8 Mackay Junior College of Medicine, Nursing and Management, Taipei, Taiwan 9 Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan 10 Department of Medicine, Mackay Medical College, New Taipei City, Taiwan 11 Department of Biological Science and Technology, National Chaio Tung University, Hsinchu, Taiwan Correspondence to: Cheng-Jui Lin, email: lincj@mmh.org.tw Keywords: indoxyl sulfate, chronic kidney disease, erythropoietin, renal anemia Received: April 27, 2017 Accepted: May 29, 2017 Published: June 28, 2017 ABSTRACT Renal anemia is a common complication in patients with advanced chronic kidney disease. In vitro studies have shown that indoxyl sulfate decreases erythropoietin production. Whether this effect is seen in vivo remains unclear. Our goal was to explore the role of indoxyl sulfate in renal anemia. We found serum indoxyl sulfate levels are significantly and negatively associated with erythropoietin levels in human. A multiple stepwise linear regression analyses after adjustment for other independent parameters revealed that free indoxyl sulfate, and total indoxyl sulfate were significantly associated with erythropoietin levels. In animal studies, erythropoietin gene and protein expression were markedly inhibited in rats with chronic kidney disease; however, this effect was significantly reversed by lowering serum indoxyl sulfate with AST-120. Indoxyl sulfate may also inhibit erythropoietin expression in animal models with chronic kidney disease. These findings further support the role of indoxyl sulfate in the development of renal anemia.

Yung-Fu Chen,1–3,* Hsuan-Hung Lin,4,* Chih-Sheng Lin,5 Battsetseg Turbat,1 Kuo-An Wang,4,6 Wei-Sheng Chung1,3,7 1Department of Healthcare Administration, 2Department of Dental Technology and Materials Science, Central Taiwan University of Science and Technology, Taichung, 3Department of Health Services Administration, China Medical University, Taichung, 4Department of Management Information Systems, Central Taiwan University of Science and Technology, Taichung, Taiwan, Republic of China; 5Department of Radiology, BenQ Medical Center, Nanjing, People’s Republic of China; 6Department of Industrial Education and Technology, National Changhua University of Education, Changhua, 7Department of Internal Medicine, Taichung Hospital, Ministry of Health and Welfare, Taichung, Taiwan, Republic of China *These authors contributed equally to this work Background: Bronchiectasis is characterized by permanent dilatation of the bronchial tree caused by recurrent airway infection and inflammation. The association of atherosclerosis and inflammation is well established. However, studies on the relationship between bronchiectasis and stroke are scant. Objective: We conducted a population-based cohort study to investigate the incidence and risk of ischemic stroke in patients with bronchiectasis. Methods: Data of 1,295 patients newly diagnosed as bronchiectasis between 2000 and 2008 were retrieved from the Taiwan National Health Insurance Research Database. A total of 6,475 controls without bronchiectasis at a ratio of 5:1 were randomly selected from the general population based on frequency-matched age and sex to the patients. All participants were followed up to the date of ischemic stroke development, censoring, or the end of 2010. The Cox proportional hazard model was used to identify the risk of ischemic stroke in patients with bronchiectasis compared with those without bronchiectasis. Results: The patients with bronchiectasis exhibited a higher incidence rate of ischemic stroke (9.18 vs 4.66 per 1,000 person-years) than the patients without bronchiectasis, with an adjusted hazard ratio of 1.74 (95% confidence interval =1.28–2.35). The patients with bronchiectasis and any comorbidities exhibited a 2.66-fold adjusted hazard ratio of ischemic stroke compared with those with neither bronchiectasis nor comorbidity (95% confidence interval =1.85–3.84). The patients with bronchiectasis carried a dose response of ischemic stroke according to the number of emergency visits and hospitalizations per year. Conclusion: This study indicated that bronchiectasis is an independent risk factor of ischemic stroke. Keywords: bronchiectasis, ischemic stroke, cohort study

Optical biosensors for rapid detection of significant foodborne pathogens are steadily gaining popularity due to its simplicity and sensitivity. While nanomaterials such as gold nanoparticles (AuNPs) are commonly used as signal amplifiers for optical biosensors, AuNPs can also be utilized as a robust biosensing platform. Many reported optical biosensors were designed for individual pathogen detection in a single assay and have high detection limit (DL). Salmonella spp. is one of the major causative agents of foodborne sickness, hospitalization and deaths. Unfortunately, there are around 2,000 serotypes of Salmonella worldwide, and rapid and simultaneous detection of multiple strains in a single assay is lacking. In this study, a comprehensive and highly sensitive simultaneous colorimetric detection of nineteen (19) environmental and outbreak Salmonella spp. strains was achieved by a novel optical biosensing platform using oligonucleotide-functionalized AuNPs. A pair of newly designed single stranded oligonucleotides (30-mer) was displayed onto the surface of AuNPs (13 nm) as detection probes to hybridize with a conserved genomic region (192-bases) of ttrRSBCA found on a broad range of Salmonella spp. strains. The sandwich hybridization (30 min, 55°C) resulted in a structural formation of highly stable oligonucleotide/AuNPs-DNA complexes which remained undisturbed even after subjecting to an increased salt concentration (2 M, final), thus allowing a direct discrimination via color change of target (red color) from non-target (purplish-blue color) reaction mixtures by direct observation using the naked eye. In food matrices (blueberries and chicken meat), nineteen different Salmonella spp. strains were concentrated using immunomagnetic separation and then simultaneously detected in a 96-well microplate by oligonucleotide-functionalized AuNPs after DNA preparation. Successful oligonucleotide/AuNPs-DNA hybridization was confirmed by gel electrophoresis while AuNPs aggregation in non-target and control reaction mixtures was verified by both spectrophotometric analysis and TEM images. Results showed that the optical AuNP biosensing platform can simultaneously screen nineteen (19) viable Salmonella spp. strains tested with 100% specificity and a superior detection limit of

To date, population-based studies on the healthcare service utilization among stable heart, kidney, and liver transplant recipients with different calcineurin inhibitors are still scarce. Therefore, we used the Taiwan National Health Insurance Research Database to conduct a nationwide cross-sectional study to estimate the healthcare utilization of stable transplant recipients with tacrolimus or cyclosporine (n = 3,482). The sampled patients in this study comprised 377 heart, 1,693 kidney, and 1,412 liver transplant recipients between 1 January 2011 and 31 December 2011. Each subject was followed for a 1-year period to evaluate his/her healthcare service utilization. Outcome variables of the healthcare service utilization were stated as below: numbers of outpatient visits, outpatient costs, numbers of inpatient days, inpatients costs, and total costs of all healthcare services. As for all healthcare service utilization, stable transplant recipients on tacrolimus had significantly more outpatient visits (40.7 vs. 38.6), outpatient costs (US$10,383 vs. US$8,155), and total costs (US$12,516 vs. US$10,372) of all healthcare services than those on cyclosporine during the 1-year follow-up period. Additionally, further analysis showed that heart transplant recipients receiving tacrolimus incurred 1.7-fold higher inpatient costs compared to patients receiving cyclosporine. We concluded that transplant recipients using tacrolimus had significantly higher utilization of all healthcare services than those receiving cyclosporine as immunosuppressive therapy.

Introduction Few studies have clinically assessed road traffic injuries in individuals with insomnia. This study examined the epidemiological characteristics of road traffic injuries in individuals with insomnia and identified relevant factors. Methods In this retrospective cohort study, we used administrative data (National Health Insurance Research Database) to identify a cohort from people admitted to hospital with insomnia and matched controls from other admitted patients. All the included data were used to determine the incidence rate of having road traffic injuries in hospital admission in individuals aged 18 and older with insomnia. A control group of individuals without insomnia with a 4:1 ratio was matched by gender, age, and index year. Cox regression models were used to evaluate the effects of insomnia on the risk of road traffic injuries (Hazard ratio, HR). The link between insomnia and repeated road traffic injury was also explored. Results From 2000 to 2015, 1,936,512 people aged 18 and older were hospitalized in Taiwan (194,035 with insomnia and 776,140 without insomnia). After other factors were controlled, the risk of road traffic injuries in individuals with insomnia was 1.23 times that of individuals without insomnia. In the 25 to 44 age group, the risk in individuals with insomnia was 1.80 times that of those without insomnia. Additionally, the risk of road traffic injuries in automobile drivers and motorcycle riders with insomnia was 1.46 and 1.29 times that of those without insomnia, respectively. The risk of repeated road traffic injuries with insomnia was 2.06 times that of those without insomnia. Conclusions To reduce the probability of road traffic injury, high-risk groups should seek early medical assistance to resolve insomnia.

Inflammation-mediated abnormalities in the renin-angiotensin system (RAS) and expression of matrix metalloproteinases (MMPs) are implicated in the pathogenesis of lung injury. Angiotensin converting enzyme II (ACE2), an angiotensin converting enzyme (ACE) homologue that displays antagonist effects on ACE/angiotensin II (Ang II) axis, could also play a protective role against lung diseases. However, the relationship between ACE2 and MMPs activation in lung injury is still largely unclear. The purpose of this study is to investigate whether MMPs activity could be affected by ACE2 and which ACE2 derived signaling pathways could be also involved via using a mouse model with lung injury induced by cigarette smoke (CS) exposure for 1 to 3 weeks. Wild-type (WT; C57BL/6) and ACE2 KO mice (ACE2(-/-)) were utilized to study CS-induced lung injury. Increases in the resting respiratory rate (RRR), pulmonary immunokines, leukocyte infiltration and bronchial hyperplasia were observed in the CS-exposed mice. Compared to WT mice, more serious physiopathological changes were found in ACE2(-/-) mice in the first week of CS exposure. CS exposure increased pulmonary ACE and ACE2 activities in WT mice, and significantly increased ACE in ACE2(-/-) mice. Furthermore, the activity of pulmonary MMPs was decreased in CS-exposed WT mice, whereas this activity was increased in ACE2(-/-) mice. CS exposure increased the pulmonary p-p38, p-JNK and p-ERK1/2 level in all mice. In ACE2(-/-) mice, a significant increase p-STAT3 signaling was detected; however, no effect was observed on the p-STAT3 level in WT mice. Our results support the hypothesis that ACE2 deficiency influences MMPs activation and STAT3 phosphorylation signaling to promote more pulmonary inflammation in the development of lung injury.

Erectile dysfunction (ED) affects millions of men worldwide. Men with ED generally complain failure to attain or maintain an adequate erection during sexual activity. The prevalence of ED is strongly correlated with age, affecting about 40% of men at age 40 and nearly 70% at age 70. A variety of chronic diseases, including diabetes, ischemic heart disease, congestive heart failure, hypertension, depression, chronic renal failure, obstructive sleep apnea, prostate disease, gout, and sleep disorder, were reported to be associated with ED. In this study, data retrieved from a subset of the National Health Insurance Research Database of Taiwan were used for designing the clinical decision support system (CDSS) for predicting ED incidences in men. The positive cases were male patients aged 20-65 who were diagnosed with ED between January 2000 and December 2010 confirmed by at least three outpatient visits or at least one inpatient visit, while the negative cases were randomly selected from the database without a history of ED and were frequency (1:1), age, and index year matched with the ED patients. Data of a total of 2832 ED patients and 2832 non-ED patients, each consisting of 41 features including index age, 10 comorbidities, and 30 other comorbidity-related variables, were retrieved for designing the predictive models. Integrated genetic algorithm and support vector machine was adopted to design the CDSSs with two experiments of independent training and testing (ITT) conducted to verify their effectiveness. In the 1st ITT experiment, data extracted from January 2000 till December 2005 (61.51%, 1742 positive cases and 1742 negative cases) were used for training and validating and the data retrieved from January 2006 till December 2010 were used for testing (38.49%), whereas in the 2nd ITT experiment, data in the training set (77.78%) were extracted from January 2000 till Deceber 2007 and those in the testing set (22.22%) were retrieved afterward. Tenfold cross validation and three different objective functions were adopted for obtaining the optimal models with best predictive performance in the training phase. The testing results show that the CDSSs achieved a predictive performance with accuracy, sensitivity, specificity, g-mean, and area under ROC curve of 74.72%–76.65%, 72.33%–83.76%, 69.54%–77.10%, 0.7468–0.7632, and 0.766–0.817, respectively. In conclusion, the CDSSs designed based on cost-sensitive objective functions as well as salient comorbidity-related features achieve satisfactory predictive performance for predicting ED incidences.

Patient readmission may be attributed to unsuccessful treatments, new diseases, or worsening comorbidities, which manifest degraded healthcare quality and increased cost of a hospital. Clinical decision support systems (CDSSs) provide useful information and expert knowledge to improve diagnostic performance, treatment outcomes, and healthcare quality in clinical settings. The objectives of this study is to design a CDSS based on the data retrieved from the National Health Insurance Research Database (NHIRD) for assisting physicians in the identification of high-risk patients of 30-day readmissions. A wrapper method integrating genetic algorithm (GA) and support vector machine (SVM) was used to designing the CDSSs by selecting salient features and designing the classifiers based on 2 under-sampling methods and 3 objective functions. The accuracy, sensitivity, specificity, and area under ROC curve (AUC) of the CDSSs achieve 69.33-71.44%, 66.27-69.41%, 69.32-72.24%, and 0.7518-0.7601, respectively, which outperform the models designed based on data retrieved from electronic medical records. An online web service system is provided to assist physicians for detecting potential pneumonia patients who have higher probability of all-cause readmission after being discharged from hospitals, thereby giving them necessary interventions to reduce morbidities and mortalities and to reduce healthcare cost.

Angiotensin-converting enzyme (ACE) is the primary enzyme that converts angiotensin I (Ang I) to angiotensin II (Ang II) in the renin-angiotensin system (RAS). However, chymase hydrates Ang I to Ang II independently of ACE in some kidney diseases, and it may play an important role. The present study investigated whether chymase played a crucial role in aristolochic acid I (AAI)-induced nephropathy. C57BL/6 mice were treated with AAI via intraperitoneal injection for an accumulated AAI dosage of 45 mg/kg body weight (BW) (15 mg/kg BW per day for 3 days). The animals were sacrificed after acute kidney injury development, and blood, urine and kidneys were harvested for biochemical and molecular assays. Mice exhibited increased serum creatinine, BUN and urinary protein after the AAI challenge. Significant infiltrating inflammatory cells and tubular atrophy were observed in the kidneys, and high immunocytokine levels were detected. Renal RAS-related enzyme activities were measured, and a significantly increased chymase activity and slightly decreased ACE activity were observed in the AAI-treated mice. The renal Ang II level reflected the altered profile of RAS enzymes and was significantly increased in AAI-treated mice. Treatment of AAI-induced nephropathic mice with an ACE inhibitor (ACEI) or chymase inhibitor (CI; chymostatin) reduced renal Ang II levels. The combination of ACEI and CI (ACEI+CI) treatment significantly reversed the AAI-induced changes of Ang II levels and kidney inflammation and injuries. AAI treatment significantly increased renal p-MEK without increasing p-STAT3 and p-Smad3 levels, and p-MEK/p-ERK1/2 signalling pathway was significantly activated. CI and ACEI+CI treatments reduced this AAI-activated signaling pathway. AAI-induced nephropathy progression was significantly mitigated with CI and ACEI+CI treatment. This study elucidates the role of RAS in the pathogenesis of AAI-induced nephropathy.

Air pollution is a very critical issue worldwide, particularly in developing countries. Particulate matter (PM) is a type of air pollution that comprises a heterogeneous mixture of different particle sizes and chemical compositions. There are various sources of fine PM (PM2.5), and the components may also have different effects on people. The pathogenesis of PM2.5 in several diseases remains to be clarified. There is a long history of epidemiological research on PM2.5 in several diseases. Numerous studies show that PM2.5 can induce a variety of chronic diseases, such as respiratory system damage, cardiovascular dysfunction, and diabetes mellitus. However, the epidemiological evidence associated with potential mechanisms in the progression of diseases need to be proved precisely through in vitro and in vivo investigations. Suggested mechanisms of PM2.5 that lead to adverse effects and chronic diseases include increasing oxidative stress, inflammatory responses, and genotoxicity. The aim of this review is to provide a brief overview of in vitro and in vivo experimental studies of PM2.5 in the progression of various diseases from the last decade. The summarized research results could provide clear information about the mechanisms and progression of PM2.5-induced disease.

More than 1 billion people suffer from chronic respiratory diseases worldwide, accounting for more than 4 million deaths annually. Inhaled corticosteroid is a popular medication for treating chronic respiratory diseases. Its side effects include decreased bone mineral density and osteoporosis. The aims of this study are to investigate the association of inhaled corticosteroids and fracture and to design a clinical support system for fracture prediction. The data of patients aged 20 years and older, who had visited healthcare centers and been prescribed with inhaled corticosteroids within 2002–2010, were retrieved from the National Health Insurance Research Database (NHIRD). After excluding patients diagnosed with hip fracture or vertebrate fractures before using inhaled corticosteroid, a total of 11645 patients receiving inhaled corticosteroid therapy were included for this study. Among them, 1134 (9.7%) were diagnosed with hip fracture or vertebrate fracture. The statistical results showed that demographic information, chronic respiratory diseases and comorbidities, and corticosteroid-related variables (cumulative dose, mean exposed daily dose, follow-up duration, and exposed duration) were significantly different between fracture and nonfracture patients. The clinical decision support systems (CDSSs) were designed with integrated genetic algorithm (GA) and support vector machine (SVM) by training and validating the models with balanced training sets obtained by random and cluster-based undersampling methods and testing with the imbalanced NHIRD dataset. Two different objective functions were adopted for obtaining optimal models with best predictive performance. The predictive performance of the CDSSs exhibits a sensitivity of 69.84–77.00% and an AUC of 0.7495–0.7590. It was concluded that long-term use of inhaled corticosteroids may induce osteoporosis and exhibit higher incidence of hip or vertebrate fractures. The accumulated dose of ICS and OCS therapies should be continuously monitored, especially for patients with older age and women after menopause, to prevent from exceeding the maximum dosage.

Background: Cyclophilin A (CyPA) concentration increases in acute coronary syndrome. In an animal model of acute myocardial infarction, administration of angiotensin-converting-enzyme inhibitor was associated with lower left ventricular (LV) CyPA concentration and improved LV performance. This study investigated the relationships between changes in plasma CyPA concentrations and LV remodeling in patients with ST-elevation myocardial infarction (STEMI). Methods and Results: We enrolled 55 patients who underwent percutaneous coronary intervention for acute STEMI. Plasma CyPA, matrix metalloproteinase (MMP), interleukin-6 and high-sensitivity C-reactive protein concentrations were measured at baseline and at one-month follow-up. Echocardiography was performed at baseline and at one-, three-, and six-month follow-up. Patients with a decrease in baseline CyPA concentration at one-month follow-up (n = 28) had a significant increase in LV ejection fraction (LVEF) (from 60.2 ± 11.5% to 64.6 ± 9.9%, p < 0. 001) and preserved LV synchrony at six months. Patients without a decrease in CyPA concentration at one month (n = 27) did not show improvement in LVEF and had a significantly increased systolic dyssynchrony index (SDI) (from 1.170 ± 0.510% to 1.637 ± 1.299%, p = 0.042) at six months. Multiple linear regression analysis showed a significant association between one-month CyPA concentration and six-month LVEF. The one-month MMP-2 concentration was positively correlated with one-month CyPA concentration and LV SDI. Conclusions: Decreased CyPA concentration at one-month follow-up after STEMI was associated with better LVEF and SDI at six months. Changes in CyPA, therefore, may be a prognosticator of patient outcome.

Background: The renin-angiotensin system (RAS) has significant influences on heart and renal disease progression. Angiotensin converting enzyme (ACE) and angiotensin converting enzyme II (ACE2) are major peptidases of RAS components and play counteracting functions through angiotensin II (Ang II)/ATIR and angiotensin-(1–7) (Ang-(1–7))/Mas axis, respectively. Methods: There were 360 uremic patients on regular hemodialysis (HD) treatment (inclusive of 119 HD patients with cardiovascular diseases (CVD) and 241 HD patients without CVD and 50 healthy subjects were enrolled in this study. Plasma ACE, ACE2, Ang II and Ang-(1–7) levels of the HD patients were determined. Results: We compared pre-HD levels of plasma ACE, ACE2, Ang II and Ang-(1–7) in the HD patients with and without CVD to those of the controls. The HD patients, particularly those with CVD, showed a significant increase in the levels of ACE and Ang II, whereas ACE2 and Ang-(1–7) levels were lower than those in the healthy controls. Therefore, imbalanced ACE/ACE2 was observed in the HD patients with CVD. In the course of a single HD session, the plasma ACE, ACE/ACE2 and Ang II levels in the HD patients with CVD were increased from pre-HD to post-HD. On the contrary, ACE2 levels were decreased after the HD session. These changes were not detected in the HD patients without CVD. Conclusions: Pathogenically imbalanced circulating ACE/ACE2 was detected in the HD patients, particularly those with CVD. HD session could increase ACE/Ang II/AT1R axis and decrease ACE2/Ang-(1–7)/Mas axis activity in the circulation of HD patients with CVD.

Pancreatitis is the inflammation of the pancreas. Chymotrypsin, an indicator of pancreatic function, could serve as a biomarker for the diagnosis of pancreatitis. A gold nanoparticle (AuNP)-based fluorescence assay was fabricated in this study to assay the activity of chymotrypsin. Peptides labeled with fluorophores were conjugated onto AuNPs as a chymotrypsin-activated peptide-functionalized AuNP probe (AuNPs-peptide probe). The detection sensitivity of the AuNPs-peptide probe toward proteolytic activity was significantly increased by using spacer-enhanced peptides with specifically designed lengths and charges. The limit of detection of the designed AuNPs-peptide probe decreased, enabling detection at the pM level within a 15 min detection time. The AuNPs-peptide probe was used to evaluate chymotrypsin activity as an indicator of acute pancreatitis (AP) in a mouse model induced by cerulein challenge. The fecal chymotrypsin activity in cerulein-induced AP mice was significantly lower than that observed in controls. This is the first study to use an AuNPs-peptide probe for the diagnosis of pancreatitis using fecal specimens.

Renal anemia is a common complication in patients with advanced chronic kidney disease.

Single-incision laparoscopic surgery developed rapidly in recent years. We introduce an innovative technique: single-incision laparoscopic common bile duct exploration (SILCBDE) with conventional instruments. A retrospective comparison between SILCBDE and standard laparoscopic common bile duct exploration (LCBDE) was analyzed.Thirty-four patients who underwent LCBDE for choledocholithiasis in a period of 17 months were enrolled. Seventeen standard LCBDEs and 17 SILCBDEs were attempted. Simultaneous cholecystectomies were performed.The stone clearance rate was 94.1% (16 patients) in the standard LCBDE group and 100% in the SILCBDE group. There was no statistical difference in demographic distribution, clinical presentations, and operative results between the two groups, except the SILCBDE group had a higher rate of acute cholecystitis than the standard LCBDE group (76.5 vs. 35.3%; p 0.05). One procedure (5.9%) in the SILCBDE group was converted to a four-incision transcystic LCBDE. The complication rate was 11.8% (two patients) in the standard LCBDE group and 5.9% (one patient) in the SILCBDE group. The average follow-up period was 4.2 months.SILCBDE is as safe and efficacious as standard LCBDE in experienced hands. Choledochoscope manipulation and bile duct repair are the key skills. Long-term follow-up and further prospective randomized trials are anticipated.

Conventional detection methods for Listeria monocytogenes contamination in food are time-consuming and expensive, creating a need for a more inexpensive and efficient detection method. The objective of this study was to develop an inexpensive biosensor to detect L. monocytogenes in food samples. Screen-printed carbon electrode (SPCE) strips previously used by diabetic patients for glucose monitoring were modified with gold nanoparticles (AuNPs) and specific antibodies to L. monocytogenes. To facilitate detection L. monocytogenes was amplified by conjugating secondary enzyme-labeled antibodies with AuNPs. This assay has the ability to detect L. monocytogenes at 2 log CFU/g in wild blueberry samples and exhibits significant specificity over other enteric pathogens such as Escherichia coli O157:H7 and Salmonella Typhimurium. These results indicate that modifying the electrodes with AuNPs is crucial in the development and production of SPCE biosensors. The use of this optimized amperometric immuno-bi...

Image retake of radiological examinations not only increases the risk of radiation exposure of the patients, but also wastes the medical resource and degrades the quality of services of the hospitals. This study aimed at discovering factors affecting image retake of general digital radiography for setting guidelines to reduce the image retaking rate. A total of 98,503 general X-ray images retrieved from the picture archiving and communication system database of a medical center in central Taiwan were analyzed. The results showed that the total retaking rate was 4.89% with the position error (56.05%) was the main factor causing image retakes and chest examination showed the highest frequency (1544 cases). On the other hand, skull/face exhibited the highest retaking rate (9.81%) among various types of examinations. After discovering the factors affecting the image retaking rate, suitable guidelines were proposed and introduced. The image retake rate had been significantly reduced to 4.38% and 3.57% 1 month and 6 months, respectively, after the introduction of guidelines. In conclusion, image retake analysis is a quality indicator and is effective for quality assurance of digital radiology. Regular analysis of image retake can find factors inducing image retake and is useful for designing guidelines to reduce the image retake rate.

Matrix metalloproteinase 9 (MMP-9), a member of MMP family, is involved in many physiological processes, including cardiovascular disease (CVD). Tumor necrosis factor-α (TNF-α) is considered a cytokine with pleiotropic biological capabilities and leads to the process of CVD when TNF-α is abnormally released and stimulates MMP-9 expression and activation. In this study, we investigated the molecular mechanism of TNF-α-regulated MMP-9 expression. The experimental results confirm that TNF-α could upregulate MMP-9 expression in heart myoblast H9c2 cells of rat. To evaluate the MMP-9 regulation at transcriptional level, a DNA fragment of 2.2 kb (-2168/+18) of human mmp-9 promoter region was cloned and constructed in a vector of luciferase reporter gene. The 2.2-kb sequences were identified as having three candidate nuclear factor-κ B (NF-κB) binding sites: NF-κB I (-1418/-1409), NF-κB II (-626/-617), and NF-κB III (-353/-345). A series of reporter vectors with the mutated NF-κB sites of mmp-9 promoter sequences were constructed and transfected into H9c2 cells. The results show that the NF-κB II binding site (-626/-617) within the promoter region of mmp-9 plays a key role in upregulation of mmp-9 expression by TNF-α induction. In addition, we also first identified that the NF-κB I, similar to c-Rel, might be one of the NF-κB families to regulate mmp-9 expression.

A sensitive bacteria enrichment and detection system for viable Escherichia coli O157:H7 was developed using a piezoelectric biosensor-quartz crystal microbalance (QCM) with antibody-functionalized gold nanoparticles (AuNPs) used as detection verifiers and amplifiers. In the circulating-flow QCM system, capture antibodies for E. coli O157:H7 were first immobilized onto the QCM chip. The sample containing E. coli O157:H7 was circulated through the system in the presence of 10 ml of brain heart infusion (BHI) broth for 18 h. The cells of E. coli O157:H7 specifically captured and enriched on the chip surface of the QCM were identified by QCM frequency changes. Listeria monocytogenes and Salmonella Typhimurium were used as negative controls. After bacterial enrichment, detection antibody-functionalized AuNPs were added to enhance the changes in detection signal. The use of BHI enrichment further enhanced the sensitivity of the developed system, achieving a detection limit of 0–1 log CFU/ml or g. The real-time monitoring method for viable E. coli O157:H7 developed in this study can be used to enrich and detect viable cells simultaneously within 24 h. The unique advantages of the system developed offer great potential in the microbial analysis of food samples in routine settings.

Objective: Pleural effusion is common problem, but the rapid and reliable diagnosis for specific pathogenic effusions are lacking. This study aimed to identify the diagnosis based on clinical variables to differentiate pleural tuberculous exudates from other pleural effusions. We also investigated the role of renin-angiotensin system (RAS) and matrix metalloproteinase (MMPs) in the pathogenesis of pleural exudates. Experimental design: The major components in RAS and extracellular matrix metabolism, including angiotensin converting enzyme (ACE), ACE2, MMP-2 and MMP-9 activities, were measured and compared in the patients with transudative (n = 45) and exudative (n = 80) effusions. The exudative effusions were come from the patients with tuberculosis (n = 20), pneumonia (n = 32), and adenocarcinoma (n = 28). Results: Increased ACE and equivalent ACE2 activities, resulting in a significantly increased ACE/ACE2 ratio in exudates, were detected compared to these values in transudates. MMP-9 activity in exudates was significantly higher than that in transudates. The significant correlation between ACE and ACE2 activity that was found in transudates was not found in exudates. Advanced analyses showed significantly increased ACE and MMP-9 activities, and decreased ACE2 activity in tuberculous pleural effusions compared with those in pneumonia and adenocarcinoma effusions. The results indicate that increased ACE and MMP-9 activities found in the exudates were mainly contributed from a higher level of both enzyme activities in the tuberculous pleural effusions. Conclusion: Interplay between ACE and ACE2, essential functions in the RAS, and abnormal regulation of MMP-9 probably play a pivotal role in the development of exudative effusions. Moreover, the ACE/ACE2 ratio combined with MMP-9 activity in pleural fluid may be potential biomarkers for diagnosing tuberculous pleurisy.

Diosgenin, a traditional Yam extraction, has been used in hormone replacement for menopausal women. We aimed to investigate the influences of diosgenin administration upon the MMP-2 and -9 activity and expression and reproductive hormones of ovariectomized (OVX) rats, a model of menopausal status. Seven-week old female Wistar rats with bilateral OVX or sham operation (controls) were divided and administered different dosages of diosgenin (0, 10, 50, or 100 mg/kg/day) for 8 weeks. Serum was then sampled for progesterone (P4) and estradiol (E2) assay and uterine horns harvested. Myometrial MMP-2 and -9 activity and expression were surveyed and myometrial collagen expression was also assayed. The results show higher body weight in OVX rats across the 8 weeks post surgery and no significant differences were noted among OVX or Sham rats with diosgenin supplements. There were lower P4 and E2 concentrations in OVX rats compared to Sham rats, and higher P4 concentration of Sham rats post diosgenin supplement. MMP-2 and -9 mRNA expression and activity was lower in OVX rats, although higher MMP-2 and lower MMP-9 activity/mRNA expression was observed in OVX rats post diosgenin supplementation. Collagen mRNA expression was higher in OVX rats compared to Sham controls, and diosgenin administration decreased collagen mRNA expression in OVX rats. In conclusion, diosgenin is associated with gelatinase expression and collagen metabolism in OVX rats. Diosgenin administration can partially reverse the effects of OVX upon MMP functions and hormone status. Adequate diosgenin supplement might modulate myometrial gelatinase expression and collagen metabolism in menopausal subjects.

Condensation Both Gonal-F and Puregon, especially in their high-dosage administration, might inhibit the endometrial cell proliferation in the initial 48-hour culture. After 72-hour culture, Gonal-F persisted the inhibition of the endometrial growth, whereas Puregon reversed its effect to enhance endometrial growth. Objectives Endometrial proliferation or regeneration during menstrual cycle is regulated by sexual hormones. However, the effect of gonadotropins on the endometrial cell growth remains obscure. Herein, we aimed to investigate the effects of recombinant follicle-stimulating hormones (r-FSHs) (Gonal-F and Puregon) on the proliferation of human endometrial cells in vitro . Materials and Methods Human endometrial cells (RL95-2 cells) were obtained commercially and cultured in the serum-containing media in the presence of r-FSHs (Gonal-F and Puregon at concentrations of 0mIU/mL, 200mIU/mL, 400mIU/mL, and 600mIU/mL) up to 72 hours. According to the gonadotropin concentrations, all cultured endometrial cells were divided into four groups: (1) 0mIU/mL (control); (2) 200mIU/mL; (3) 400mIU/mL; and (4) 600mIU/mL. After 72-hour culture, endometrial cell proliferations were assessed overnight by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The influences of different r-FSH agents and dosages on endometrial cell proliferation in each group were evaluated and compared. Results In the four Gonal-F groups, the cell absorption (control and 200mIU/mL, 400mIU/mL, and 600mIU/mL Gonal-F) after 24/48/72-hour cultures were as follows: (1) 0.57/0.7/0.82; (2) 0.56/0.66/0.78; (3) 0.55/0.64/0.77; and (4) 0.51/0.61/0.78. After 48 hours, higher dosage of Gonal-F appeared to significantly inhibit the endometrial cell proliferation. After 72-hour culture, all three dosages of Gonal-F appeared to inhibit the endometrial cell proliferation similarly. In Puregon groups, the cell absorptions were as follows: (1) 0.62/0.53/0.62; (2) 0.61/0.5/0.66; (3) 0.61/0.49/0.66; and (4) 0.64/0.49/0.66. Puregon administration displayed initial inhibition and subsequent stimulation effects on the endometrial cells. Conclusions Both Gonal-F and Puregon, especially in their high-dosage administration, appeared to inhibit the endometrial cell proliferation in the initial 48-hour culture. After 72-hour culture, Gonal-F persisted the inhibition of the endometrium, whereas Puregon reversed its effect by enhancing the endometrial growth. The differences might be because of the different formulations or molecular structures existing between alpha and beta follitropins.

Summary Objective We aimed to investigate the effects of arsenic (As), benomyl (Ben), and carbendazim (Carb) on endometrial cells. Materials and Methods Human endometrial cells were obtained during diagnostic curettage. All cultured endometrial cells were divided into four groups: (1) 0 M (controls), (2) 10 −6 M, (3) 10 −5 M, (4) 10 −4 M for As, Ben and Carb. After 24 and 48 hours in culture, endometrial cell proliferations were assessed by diphenyltetrazolium bromide assay. The influences of different concentrations of As, Ben and Carb upon the endometrium were compared. Results During the first 24 hours, As, Ben and Carb appeared to have insignificant influences upon endometrial growth. After 48 hours in culture, all three agents significantly inhibited endometrial growth. In As groups, cell absorption after 48 hours culture were 100% (group 1), 82.1% (group 2), 43.6% (group 3) and 35.3% (group 4). In Ben groups, cell absorption was 100% (1), 75.9% (2), 66.4% (3) and 49. 6% (4). In the Carb groups, cell absorption was 100% (1), 70.4% (2), 73.0% (3) and 76.7% (4). Conclusion The agents As, Ben and Carb appear to have inhibitory effects upon endometrial cells after 48 hours in culture.

Numerous studies have suggested that angiotensin peptides modulate the expression of angiotensin converting enzyme II (ACE2) in the cardiovascular system, but the molecular mechanisms remain poorly understood. In the present study, human cardiac fibroblasts (HCF) were used to test the regulatory effects of angiotensin II (Ang II) and angiotensin-(1-7) [Ang-(1-7)] on ACE2 expression. The results show that Ang II upregulates ACE2 expression. This action is modulated through activation of Ang II type 1 receptor (AT1R). Ang II-mediated ACE2 upregulation was blocked by antagonists of AT1R and ERK-MAPK signaling pathways. Additionally, Ang-(1-7) increased ACE2 expression, and this upregulation was inhibited by Ang-(1-7) Mas receptor blockade. Our results further reveal that the activation of p-ERK1/2 proteins plays a critical role in upregulating ACE2 in Ang-(1-7)-stimulated HCF cells. This effect occurs independently of the Ang II-AT1R signaling pathway. In conclusion, we propose that Ang II-upregulated ACE2 may increase Ang-(1-7) formation from Ang II, and that ACE2 expression is further enhanced by Ang-(1-7) in a positive feedback loop.

Rabbit neutrophil peptide-1 (NP-1) is a prototypic rabbit α-defensin with a broad antimicrobial spectrum. The coding sequence of NP-1 was amplified and cloned into pET-31b(+) to construct an expression vector, pET31-NP1, which was transformed into E. coli BLR(DE3)pLysS for expressing the fusion NP-1 protein (fNP-1). The fNP-1 is downstream of a ketosteroid isomerase (KSI) and upstream of a (His)6-Tag, as KSI-NP1-His6. The optimal condition, cultivation in enriched LB medium and induction with 0.5 mM IPTG for 6 h, was determined for fNP-1 production. The fNP-1 was purified by Ni-NTA resin and cleaved by cyanogen bromide to release matured NP-1 peptide. The matured NP-1 peptide showed significant antimicrobial activities against clinical bacteria, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus. The application of this expression approach represents a potential method to produce NP-1 by fusion protein expressed in E. coli without cytotoxicity.

Hirudin, isolated from the leech Hirudo medicinalis, inhibits thrombin directly and several expression systems have been used to produce recombinant Hirudin (rHirudin) for pharmaceutical purposes. A DNA fragment containing the Hirudin coding sequence and goat beta-casein secretion signal was chemically synthesized in this study. The synthetic DNA then was further constructed into a goat beta-casein expression vector for mouse transgenesis. Four lines of transgenic mice were successfully developed and one line showed a meaningful anti-thrombin activity of 40,000 anti-thrombin units (ATU)/mL in their milk. In this animal line, Hirudin mRNA was found in samples of uterus and kidney with insignificant anti-thrombin activity (

A disposable amperometric immunosensing strip was fabricated for rapid detection of Escherichia coli O157:H7. The method uses an indirect sandwich enzyme-linked immunoassay with double antibodies. Screen-printed carbon electrodes (SPCEs) were framed by commercial silver and carbon inks. For electrochemical characterization the carbon electrodes were coupled with the first E. coli O157:H7-specific antibody, E. coli O157:H7 intact cells and the second E. coli O157:H7-specific antibody conjugated with horseradish peroxidase (HRP). Hydrogen peroxide and ferrocenedicarboxylic acid (FeDC) were used as the substrate for HRP and mediator, respectively, at a potential +300 mV vs. counter/reference electrode. The response current (RC) of the immunosensing strips could be amplified significantly by 13-nm diameter Au nanoparticles (AuNPs) attached to the working electrode. The results show that the combined effects of AuNPs and FeDC enhanced RC by 13.1-fold. The SPCE immunosensing strips were used to detect E. coli O157:H7 specifically. Concentrations of E. coli O157:H7 from 102 to 107 CFU/ml could be detected. The detection limit was approximately 6 CFU/strip in PBS buffer and 50 CFU/strip in milk. The SPCE modified with AuNPs and FeDC has the potential for further applications and provides the basis for incorporating the method into an integrated system for rapid pathogen detection.

Angiotensin II (Ang II) is a critical effector in the renin-angiotensin system (RAS), which modulates cardiovascular homeostasis, and the matrix metalloproteinases (MMPs) and tissue inhibitors of matrix metalloproteinases (TIMPs) related metabolism of extracellular matrix (ECM). Angiotensin(1-7) [Ang(1-7)] is another bioactive peptide in the RAS and is considered to have opposite effects to Ang II. However, the modulation of MMPs and TIMPs by Ang(1-7) is largely unclear in cardiocytes, and the antagonistic effects of Ang(1-7) on Ang II-mediated expression of MMPs and TIMPs have yet to be identified. In the present study, we examined the transcript expression of MMPs and TIMPs in human cardiac fibroblasts (HCF) and myocytes (HCM) after Ang II or Ang(1-7) stimulation, and analysed the antagonistic effects of Ang(1-7) to Ang II. The results show that Ang II decreased transcript expression of MMP-1, MMP-2, TIMP-1, TIMP-2 and TIMP-3, but upregulated MMP-9 expression in the HCF cells. Transcript expression of MMP-9 and TIMP-2 was downregulated by Ang(1-7) in the same cells. In the HCM cells, Ang II induced MMP-1 and MMP-9 overexpression but MMP-2 was downregulated. All of the examined MMPs and TIMPs, except MMP-9, were markedly decreased by Ang(1-7). In the studies of antagonistic effects of Ang(1-7) to Ang II, Ang(1-7) counteracted the effects of Ang II-mediated regulation on MMP-9 and TIMP-1 in the HCF cells compared with the control group. The regulations of all examined MMPs by Ang II were reversed to basal expression by Ang(1-7) in the HCM cells. Our results suggest that Ang(1-7) and Ang II have opposite and antagonistic effects on regulation of transcription of MMPs and TIMPs in primary cultures of human cardiocytes. These effects lead to increased ratios of MMPs to TIMPs after Ang II stimulation and decreased ratios of MMPs to TIMPs after Ang(1-7) stimulation; effects which may partly depend of the type of cardiac cells. These results suggest a potential role for Ang(1-7) in attenuatating cardiac damage in Ang II-induced ECM remodelling.

Matrix metalloproteinases (MMPs) and their endogenous tissue inhibitors (TIMPs) play important roles in the pathophysiology of renal diseases. Imbalanced MMPs/TIMPs are implicated in the vascular alterations of uremic patients on hemodialysis (HD). We have investigated the plasma levels of MMP-2, MMP-9, TIMP-1 and TIMP-2 in uremic patients and the effects of a course of HD on the changes in these factors. There were 382 uremic patients on regular HD treatment and 50 healthy controls enrolled in this study. The plasma MMP-2 and MMP-9 levels were detected by gelatin zymography, and TIMP-1 and TIMP-2 concentrations were determined by ELISA assay. Significantly higher plasma MMP-2 and MMP-9 and decreased TIMP-1 in the uremic patients were detected compared with those in the controls. Therefore, there were markedly higher MMP-2/TIMP-2 and MMP-9/TIMP-1 ratios in the uremic patients. In the course of a single HD session, the plasma MMP-2 level was significantly decreased from pre-HD to post-HD. TIMP-1 concentration was significantly increased from pre-HD to post-HD. Although the HD session did not have a significant effect on the levels of plasma MMP-9 and TIMP-2, both plasma MMP-2/TIMP-2 and MMP-9/TIMP-1 ratios were significantly decreased from pre-HD to post-HD levels. HD session could decrease MMP-2 and increase TIMP-1 level in the circulation of uremic patients. The physiological significance of reduced MMPs/TIMPs ratio due to a single HD session is required to further validate.

We have established a spontaneously immortalized porcine mammary epithelial cell line (SI-PMEC) from the mammary gland of a lactating sow and maintained it long-term in culture by continuous subculturing. SI-PMEC cells were maintained for more than 8 months (70 passages) in DMEM/F12 medium supplemented with 10% fetal calf serum, insulin, and hydrocortisone without obvious signs of senescence. When grown at low density on a plastic substratum, SI-PMEC cells formed islands, and when grown to confluency, the cells formed a monolayer and aggregated with the characteristic cobblestone morphology of epithelial cells. The subcultured SI-PMEC cells appeared to proliferate without changes in morphology or growth pattern, with an estimated population doubling time of 20-22 h. With increasing density, SI-PMEC cells organized into lumen-like structures with elongated cells at the margins. SI-PMEC cells from stocks frozen at Passage 30 were subcultured up to 20 times without changes in cell viability, proliferation rate, or morphology. Furthermore, SI-PMEC cells remained immunopositive to an antibody against cytokeratin AE3 and immunonegative to an antibody against a human fibroblast antigen. The SI-PMEC cells could form functional structures resembling ducts, lateral-buds, and alveoli in a Matrigel matrix-dependent manner in vitro. When grown on the Matrigel and stimulated by prolactin, the cells differentiated and formed mammary gland structures and strongly expressed transcripts encoding the milk proteins alpha-lactalbumin, beta-casein, and beta-lactoglobulin. Our results indicate that the SI-PMEC cell line can be subpassaged many times and still form functional differentiated secretory structures. To our knowledge, this is the first report of an immortalized mammary epithelial cell line from pig.

Objectives: Reactive oxygen species (ROS) induced lipid peroxidation is associated with sperm function. Malondialdehyde (MDA) concentration and glutathione peroxidase (GPx) activity represent the lipid peroxidation and spermicidal antioxidant, respectively. We aimed to evaluate the relationship of MDA and GPx levels with sperm parameters. Patients and methods: Specimens were divided into two groups: group 1. normospermia (n=20); group 2. oligoasthenospermia (n=31). Seminal MDA concentration was measured by thiobarbituric acid reaction method. Seminal GPx activity was measured by oxidation of reduced nicotinamide-adenine dinucleotide. Seminal MDA levels and GPx activities in both groups were compared. Results: MDA concentrations in both groups were significantly different (1.52 ± 0.75 vs. 2.25 ± 0.88 nM, p = 0.0021). GPx activities in both groups were non-significantly different (0.48 ± 0.11 vs. 0.47 ± 0.12 U/ml). MDA levels were negatively correlated with the sperm motility (MDA = -0.014 x motility + 2.62, p =0.017) and concentration (MDA = -0.0045 x concentration + 2.23, p = 0.0166). GPx activities were positively but non-significantly correlated with the sperm concentration and sperm motility. Conclusions: Seminal MDA concentrations are negatively correlated with sperm concentration and motility, which might provide a simple and useful tool in predicting sperm parameters. GPx activity is non-significantly correlated with the seminal quality. Roles of seminal MDA upon spermatogenesis merits further surveys.

The objective of this study was to compare gene transcription profiles of LM between two pig breeds, Duroc and Taoyuan, which display dramatically different postnatal muscle growth. We isolated LM from neonatal pigs, and the Duroc muscle length and mass were greater (P < 0.01) than for Taoyuan pigs; however, insignificant differences in the muscle fiber area and the percentage of fiber types were found. A human high-density complementary DNA (cDNA) microarray consisting of 9,182 probes was used to compare gene transcription profiles of LM between the two breeds. The results showed that the transcription level of 73 genes and 44 genes in Duroc LM were upregulated and downregulated by at least 1.75-fold (P < 0.05) compared with Taoyuan, respectively. The strongly upregulated genes in Duroc pigs included those encoding the complex of myofibrillar proteins (e.g., myosin light and heavy chains, and troponin), ribosomal proteins, transcription regulatory proteins (e.g., skeletal muscle LIM protein 1 [SLIM1] and high-mobility group proteins), and energy metabolic enzymes (e.g., electron-transferring flavo-protein dehydrogenase, NADH dehydrogenase, malate dehydrogenase, and ATP synthases). The highly transcribed genes that encode energy metabolic enzymes indicate a more glycolytic metabolism in Duroc LM, thereby favoring carbohydrates rather than lipids for use as energy substrates in this tissue. The over-transcribed genes that encode skeletal muscle-predominant proteins or transcription regulators that control myo-genesis and/or muscle growth suggest a general mechanism for the observed higher rate of postnatal muscle growth in Duroc pigs. The transcription of one such gene, SLIM1, was more highly transcribed (P < 0.01) in Duroc LM at birth and at postnatal d 7 than in Taoyuan. The transcription of SLIM1 increased (P < 0.05) in Duroc LM from neonate through 7 d of age, whereas its transcription remained essentially constant in Taoyuan during this period. These results suggest that SLIM1 may be useful for the development of markers associated with the postnatal muscle growth of pigs.