Genetic diversity and natural selection on the thrombospondin-related adhesive protein (TRAP) gene of Plasmodium falciparum on Bioko Island, Equatorial Guinea and global comparative analysis.

BACKGROUND: Thrombospondin-related adhesive protein (TRAP) is a transmembrane protein that plays a crucial role during the invasion of Plasmodium falciparum into liver cells. As a potential malaria vaccine candidate, the genetic diversity and natural selection of PfTRAP was assessed and the global PfTRAP polymorphism pattern was described. METHODS: 153 blood spot samples from Bioko malaria patients were collected during 2016-2018 and the target TRAP gene was amplified. Together with the sequences from database, nucleotide diversity and natural selection analysis, and the structural prediction were preformed using bioinformatical tools. RESULTS: A total of 119 Bioko PfTRAP sequences were amplified successfully. On Bioko Island, PfTRAP shows its high degree of genetic diversity and heterogeneity, with π value for 0.01046 and Hd for 0.99. The value of dN-dS (6.2231, p < 0.05) hinted at natural selection of PfTRAP on Bioko Island. Globally, the African PfTRAPs showed more diverse than the Asian ones, and significant genetic differentiation was discovered by the fixation index between African and Asian countries (Fst > 0.15, p < 0.05). 667 Asian isolates clustered in 136 haplotypes and 739 African isolates clustered in 528 haplotypes by network analysis. The mutations I116T, L221I, Y128F, G228V and P299S were predicted as probably damaging by PolyPhen online service, while mutations L49V, R285G, R285S, P299S and K421N would lead to a significant increase of free energy difference (ΔΔG > 1) indicated a destabilization of protein structure. CONCLUSIONS: Evidences in the present investigation supported that PfTRAP gene from Bioko Island and other malaria endemic countries is highly polymorphic (especially at T cell epitopes), which provided the genetic information background for developing an PfTRAP-based universal effective vaccine. Moreover, some mutations have been shown to be detrimental to the protein structure or function and deserve further study and continuous monitoring.

Newly excysted juveniles of Fasciola gigantica trigger the release of water buffalo neutrophil extracellular traps in vitro.

Polymorphonuclear neutrophils (PMNs) are the most abundant leukocytes and are among the first line of immune system defense. PMNs can form neutrophil extracellular traps (NETs) in response to some pathogens. The release of NETs plays an important role in trapping and killing invading parasites. However, the effects of NETs on parasitic trematode infections remain unclear. In the present study, water buffalo NET formation, triggered by the newly excysted juveniles (NEJs) of Fasciola gigantica, was visualized by scanning electron microscopy. The major components of the structure of NETs were characterized by immunofluorescence. Viability of flukes incubated with water buffalo PMNs were examined under light microscopy. The results revealed that F. gigantic juveniles triggered PMN-mediated NETs. These NETs were confirmed to comprise the classic characteristics of NETs: DNA, histones, myeloperoxidase and neutrophil elastase. Although NETs were formed in response to viable larvae, the larvae were not killed in vitro. These results suggest that NET formation may serve as a mechanism to hamper the migration of large larvae to facilitate immune cells to kill them. This study demonstrates, for the first time, that parasitic trematode juveniles can trigger NET formation.

Nasogastric tube decompression is unnecessary in patients undergoing laparoscopic nephroureterectomy for localized upper tract urothelial carcinoma.

BACKGROUND/PURPOSE: This study investigates the safety and feasibility to perform laparoscopic nephroureterectomy (LNU) for upper tract urothelial carcinoma (UTUC) without routine nasogastric tube (NGT) decompression. METHODS: The hospital-based samples comprised of 100 consecutive UTUC patients receiving elective LNU performed by two experienced surgeons. The nationwide data was based on LHID2005 composed of one million beneficiaries randomly selected from the Taiwan National Health Insurance Research Database to identify patients with the diagnoses of UTUCs receiving LNUs. We then compared baseline characteristics, peri-operative data, convalescence parameters and complications between two groups stratified by use of NGT tube. RESULTS: The hospital-based samples composed of 50 subjects with NGT and 50 without. There were no significant differences in baseline characteristics between two groups. Peri-operative and convalescence parameters were similar when comparing no NGT versus NGT: blood loss of 206 vs. 165 mL; operative time of 180.5 vs.181.1 min; days to intake was 2.1 vs.1.7 days; and hospital stay of 7.8 vs. 7.5 days (all p > 0.05). The nationwide study samples comprised 140 subjects, of which 72 were with NGT and 68 were with no NGT. The baseline data, complications and length of hospital stay were similar between two groups. CONCLUSION: Surgery-naïve patients with localized UTUC received LNU without peri-operative NGT is safe and feasible.

[Scanning Electron Microscopic Observation on Adult Gnathostoma doloresi Worms and the Phylogenetic Analysis of G. doloresi Based on ITS2 and COX1 Gene Sequences].

OBJECTIVE: To observe the ultrastructure of adult Gnathostoma doloresi worms isolated from wild boar by using scanning electron microscope (SEM), and analyze its phylogenetic relationships based on ITS2 and COXI gene sequences. METHODS: Two adult G. doloresi worms were fixed by glutaraldehyde and osmium peroxide. Ultrastructural characters of those samples were observed under SEM. Amplification and sequencing of the ITS2 and COXI genes were performed following the extraction of total genomic DNA. Sequence analysis was performed based on multiple alignments and phylogenetic analysis was made by Neighbor-Joining method using MEGA 6.0. RESULTS: The bottle-shaped adult worm covered with numerous small spines. The cervical groove connected head bulb and body without spines. There was obvious distinction in body spines which surround cervical papillae and swollen area in the middle part of the body. The fragments of ITS2 (418 bp) and COXI (381 bp) gene were obtained by PCR combined with sequencing, and were registered to the GenBank database with the accession No. of JN408329 and JN408299, respectively. The BLAST results showed that, two sequences had 99% and 98% consistency with G. doloresi ITS2 (GenBank accession No. AB181156) and COX1 (No. AB180100) gene sequences, respectively. The phylogenetic tree indicated that the two G. doloresi worms were at the same clade with a bootstrap value at 100% and 85% based on the ITS2 and COXI sequences, respectively. G. doloresi and G. hispidum were also clustered together. CONCLUSION: The results provide adequate information for the SEM morphological data of adult G. doloresi worms, and its phylogenetic relationship.

[Establishment of Duplex PCR for Identifying Metagonimus yokogawai and Haplorchis taichui].

OBJECTIVE: To develop a duplex PCR method for identifying Metagonimus yokogawai and Haplorchis taichui. METHODS: ITS1 sequences of M. yokogawai and H. taichui, as well as those of their homologous species were obtained from GenBank, and two sets of specific primer pairs for M. yokogawai and H. taichui were designed accordingly using Primer Premier 5.0 software. PCR reaction system and conditions were optimized. The established duplex PCR method was applied in a pool of M. yokogawai, H. taichui, and 17 related species to examine its specificity. Sensitivity was evaluated through serial dilutions of plasmids containing their specific sequences. Finally, the duplex PCR was applied to identify M. yokogawai and H. taichui among trematodes collected from the viscera of 47 cats and 40 dogs to test its practicality. RESULTS: The duplex PCR method amplified target sequences of M. yokogawai and H. taichui, generating 648 bp and 279 bp products, respectively. No cross reaction was found with the following 17 related species: Haplorchis pumilio, Clonorchis sinensis, Pharyngostomum cordatum, the metacercaria of Metorchis sp. and Exorchis sp., Echinochasmus liliputanus, Echinochasmus perfoliatus, Echinostoma friedi, Hypoderaeum conoideum, Holostephanus sp., Diplodiscus sp., Anisakis sp., Metorchis orientais, Paragonimus westermani, Watsonius watsoni, Notocotylus sp. and Hysterothylacium sp, indicating a high specificity of this method. The detection limits for DNAs of M. yokogawai and H. taichui were 1.49 x 10(-1) pg and 1.14 x 10(-1) pg, suggesting a good sensitivity for this method. Further, the duplex PCR successfully identified M. yokogawai and H. taichui from cat and dog viscera, with no cross amplification of other trematodes. CONCLUSION: The duplex PCR is effective in identifying Metagonimus yokogawai and Haplorchis taichui.

Factors related to the time to cryptorchidism surgery--a nationwide, population-based study in Taiwan.

BACKGROUND/PURPOSE: Current guidelines recommend that the optimal timing for cryptorchidism surgery is by the age of 12 months. This study investigated the trend of surgical timing and examined the factors associated with time to surgery for cryptorchidism in Taiwan by using a nationwide, population-based database. METHODS: The present study utilized the Longitudinal Health Insurance Database 2005, a subset of the National Health Insurance Research Database, which contains data on all paid medical benefit claims over the period 1997-2007 for a subset of 1 million beneficiaries randomly drawn from 22.72 million individuals enrolled in the National Health Insurance program in 2005. We analyzed the timing of surgery in boys younger than 18 years with diagnosis of cryptorchidism. RESULTS: We identified 547 boys who underwent surgery under 18 years of age. Approximately 79.2% of study participants received surgery after the age of 12 months. A multivariate analysis showed that several factors were significantly associated with time to surgery: age of the physician making the diagnosis, age of the surgeon performing the surgery, age of the patient at the first diagnosis of cryptorchidism, and number of previous clinic visits with the diagnosis of cryptorchidism and urbanization level of the patient's residence. CONCLUSION: A surprisingly high rate (79.2%) of all study participants underwent surgery beyond the optimal timing. Certain doctor and patient factors were associated with time to cryptorchidism surgery. Improving the alertness and education of parents and specialists may lead to earlier surgeries.

Mutations in the Cyp51A gene and susceptibility to itraconazole in Aspergillus fumigatus isolated from avian farms in France and China.

Azole resistance in the fungal pathogen Aspergillus fumigatus is an emerging problem and may develop during azole therapy in humans and animals or exposure to azole fungicides in the environment. To assess the potential risk of azole-resistance emergence in avian farms where azole compounds are used for the control of avian mycoses, we conducted a drug susceptibility study including A. fumigatus isolates from birds and avian farms in France and Southern China. A total number of 175 isolates were analyzed: 57 isolates were collected in France in avian farms where chemoprophylaxis with parconazole was performed; 51 isolates were collected in southern China in avian farms where no chemoprophylaxis was performed; and 67 additional isolates came from the collection of a mycology laboratory. No resistant isolate was detected, and the distribution of minimum inhibitory concentrations was similar for isolates collected in farms with or without azole chemoprophylaxis. For 61 randomly selected isolates, the full coding sequence of the Cyp51A gene was determined to detect mutations. Nine amino acid alterations were found in the target enzyme, 3 of which were new.

The association between ambient air pollution exposure and connective tissue sarcoma risk: a nested case-control study using a nationwide population-based database.

A nationwide population-based database was utilized in a nested case-control study to explore the association between ambient air pollution exposure and the likelihood of developing connective tissue sarcoma. The study examined 280 cases of connective tissue sarcoma diagnosed between 2000 and 2012. A random sample of 1120 control subjects was selected from a subpopulation of claim records without a connective tissue sarcoma diagnosis in a 1:4 ratio. The control subjects were selected based on similar characteristics as the connective tissue sarcoma patients, including gender, birth year, and the year of diagnosis of the case group with medical records. Risk factors for connective tissue sarcoma were collected for analysis. Our data on exposure to air pollutants was collected from Taiwan's Air Quality Monitoring Network, which has been gathering air quality data from a growing network of sampling stations (now 76) throughout the country since 1997. It was discovered that the risk of connective tissue sarcoma was significantly increased by the Charlson comorbidity index (CCI), elevated levels of specific air pollution indices (e.g., total hydrocarbons (THC), fine particulate matter (PM2.5), and O3_8 (the annual mean of the daily maximum 8-h average concentration of O3), the High Pollutant Standards Index (hPSI) (the percentage of days in a given year in Taiwan where the PSI exceeds 100), and an insurable monthly wage over US$1100. Further investigation is needed to explore the involvement of these air pollutants in the formation of connective tissue sarcoma.

RT-RPA-PfAgo detection platform for one-tube simultaneous typing diagnosis of human respiratory syncytial virus.

Human respiratory syncytial virus (HRSV) is the most prevalent pathogen contributing to acute respiratory tract infections (ARTI) in infants and young children and can lead to significant financial and medical costs. Here, we developed a simultaneous, dual-gene and ultrasensitive detection system for typing HRSV within 60 minutes that needs only minimum laboratory support. Briefly, multiplex integrating reverse transcription-recombinase polymerase amplification (RT-RPA) was performed with viral RNA extracted from nasopharyngeal swabs as a template for the amplification of the specific regions of subtypes A (HRSVA) and B (HRSVB) of HRSV. Next, the Pyrococcus furiosus Argonaute (PfAgo) protein utilizes small 5'-phosphorylated DNA guides to cleave target sequences and produce fluorophore signals (FAM and ROX). Compared with the traditional gold standard (RT-qPCR) and direct immunofluorescence assay (DFA), this method has the additional advantages of easy operation, efficiency and sensitivity, with a limit of detection (LOD) of 1 copy/µL. In terms of clinical sample validation, the diagnostic accuracy of the method for determining the HRSVA and HRSVB infection was greater than 95%. This technique provides a reliable point-of-care (POC) testing for the diagnosis of HRSV-induced ARTI in children and for outbreak management, especially in resource-limited settings.

Epidemiology of upper urinary tract stone disease in a Taiwanese population: a nationwide, population based study.

PURPOSE: We investigated the epidemiology of upper urinary tract stone disease in Taiwan using a nationwide, population based database. MATERIALS AND METHODS: This study was based on the National Health Insurance Research Database of Taiwan, which contains data on all medical beneficiary claims from 22.72 million enrollees, accounting for almost 99% of the Taiwanese population. The Longitudinal Health Insurance Database 2005, a subset of the National Health Insurance Research Database, contains data on all medical benefit claims from 1997 through 2010 for a subset of 1 million beneficiaries randomly sampled from the 2005 enrollment file. For epidemiological analysis we selected subjects whose claims records included the diagnosis of upper urinary tract urolithiasis. RESULTS: The age adjusted rate of medical care visits for upper urinary tract urolithiasis decreased by 6.5% from 1,367/100,000 subjects in 1998 to 1,278/100,000 in 2010. There was a significantly decreasing trend during the 13-year period in visits from female and all subjects (r(2) = 0.86, p = 0.001 and r(2) = 0.52, p = 0.005, respectively). In contrast, an increasing trend was noted for male subjects (r(2) = 0.45, p = 0.012). The age adjusted prevalence in 2010 was 9.01%, 5.79% and 7.38% in male, female and all subjects, respectively. The overall recurrence rate at 1 and 5 years was 6.12% and 34.71%, respectively. Male subjects had a higher recurrence rate than female subjects. CONCLUSIONS: Our study provides important information on the epidemiology of upper urinary tract stone disease in Taiwan, helping to quantify the burden of urolithiasis and establish strategies to decrease the risk of urolithiasis.

The incidence rate and characteristics in patients with testicular torsion: a nationwide, population-based study.

AIM: To determine the incidence rate and characteristics in patients with testicular torsion in Taiwan using a nationwide insurance database. METHODS: This study utilizes a subset of national health insurance research database, which contains the data of all paid medical benefit claims over the period 1997-2010 for in 1 000 000 beneficiaries in 2005. We analysed claims data for all male subjects younger than 25 years with the diagnosis of testicular torsion. RESULTS: A total of 86 subjects younger than 25 years with the diagnosis of testicular torsion were included. Among them, 22 (25.6%) underwent orchiectomies and 64 (74.4%) underwent orchiopexies. The estimated incidence of testicular torsion was 3.5 per 100 000 person-years. There are two peaks in the age-specific incidence rates: the first in boys aged 10-14 years (at 7.7 per 100 000 person-years) and the second in male infants aged <1 year (at 7.6 per 100 000 person-years). There was significant difference between orchiectomy and orchiopexy groups in the Insurance fee (p = 0.032). CONCLUSION: The incidence of testicular torsion in Taiwan was similar with previous report in the United States. It is important to improve the medical access to achieve better outcomes of testicular torsion.

Factors associated with delayed pediatric hypospadias surgery in Taiwan: a population-based, nationwide analysis.

BACKGROUND/PURPOSE: Current guidelines recommend that hypospadias repair should be performed before age 18 months. This study aims to investigate the trends of surgical timing and to determine what factors are associated with age at surgery. METHODS: The present study utilized a subset of the National Health Insurance Research Database, known as Longitudinal Health Insurance Database 2005, which contains the data of all paid medical benefit claims over the period from 1997 to 2007 for a subset of one million beneficiaries randomly drawn from the population of 22.72 million individuals in NHI program during any part of calendar year 2005. We analyzed claims data for all subjects with the diagnoses of hypospadias. RESULTS: Among 52,705 live male newborns, 218 were diagnosed with hypospadias and thus were included as subjects in our study. Among them, 89 received repair surgery. Approximately 60.6% of the study subjects received repair after the age of 18 months. Multivariate analysis showed that several factors were significantly associated with age at hypospadias surgery: specialty of clinics where first diagnosis was made; specialty of physician making the first diagnosis, age of physician making the first diagnosis; specialty of surgeon performing the surgery; number of years since surgeon's board certification; urbanization level of subject's residence; modality of surgery; concomitant cryptorchidism; concomitant prematurity and low birth weight; age at diagnosis; and number of well-baby clinic visits. CONCLUSION: This study addresses an important issue of delayed hypospadias surgery in Taiwan, which provides a potential opportunity for improvement in quality of care.

[Curcumin attenuates contrast-induced nephropathy by upregulating heme oxygenase-1 expression in rat].

OBJECTIVE: To explore the effect of curcumin (CMN) on contrast-induced nephropathy (CIN) in rats and explore the potential mechanisms focusing on heme oxygenase-1 (HO-1) expression. METHODS: Male SD rats (n = 24) were randomly divided into four groups (n = 6 each): control group (group A), diatrizoate group (DTZ, group B), DTZ + CMN group (group C), DTZ + CMN + zinc protoporphyrin IX group (group D). All rats were fed with normal chow for 1 week, right kidney was excised under anesthesia and rats were fed with normal chow for another 4 weeks. Afterwards, rats in group A was fed with normal chow, and rats in group B to D were fed with low-salt diet. All rats were injected furosemide 2 mg×kg(-1)×d(-1) for 7 days intramuscularly. At the beginning of the 7(th) day, rats in group C were injected intramuscularly with CMN 20 mg/kg, rats in group D were injected with CMN (20 mg/kg) + zinc protoporphyrin IX (7.5 mg/kg) while rats in group A and B were injected with equal volume of physiological saline. At the end of the 7(th) day, indometacin (10 mg/kg) was injected into tail vein of all rats. One hour later, 60% DTZ (8 ml/kg) was injected to rats in the group B, C and D while equal volume saline was injected to rats in group A through common carotid artery. After 48 hours, blood was drawn from the hearts of deeply anesthetized rats and kidney tissue was obtained for histology, HO-1, Bax, Bcl-2 expression and the apoptotic index measurements. RESULTS: The serum creatinine of group B, C and D [(83.67 ± 4.50) µmol/L, (63.67 ± 4.76) µmol/L, (104.17 ± 4.58) µmol/L] was significantly higher than that of group A [(41.50 ± 5.58) µmol/L, all P < 0.05], the serum creatinine was significantly higher in group B than in group C and lower than in group D (all P < 0.05). HO-1 expression of group B, C and D was significantly higher than that of group A (all P < 0.05), significantly higher in group C than in group B and D (all P < 0.05). HO-1 activity of group A, B and C was significantly higher than that of group D(all P < 0.05), HO-1 activity was significantly higher in group B than in group A and significantly lower in group B than in group C (all P < 0.05). Bax, Bcl-2 expression and apoptosis index of group B, C and D were significantly higher than that of group A (all P < 0.05), while Bcl-2/Bax of group B, C and D were significantly lower than that of group A (all P < 0.05). Bcl-2 and Bcl-2/Bax were significantly higher while apoptosis index was significantly lower in group C than in group B (all P < 0.05). Bax and apoptosis index were significantly higher and Bcl-2, Bcl-2/Bax were significantly lower in group D than in group B (all P < 0.05). CONCLUSION: CMN could protect against contrast-induced nephropathy through reducing renal cell apoptosis via upregulating HO-1 expression and activating HO-1 activity in rats.

Proteomic analysis of exosome-like vesicles from Fasciola gigantica adult worm provides support for new vaccine targets against fascioliasis.

BACKGROUND: Extracellular vesicles (EVs) released by helminths play an important role in parasite-host communication. However, little is known about the characteristics and contents of the EVs of Fasciola gigantica, a parasitic flatworm that causes tropical fascioliasis. A better understanding of EVs released by F. gigantica will help elucidate the mechanism of F. gigantica-host interaction and facilitate the search for new vaccine candidates for the control and treatment of fascioliasis. METHODS: Two different populations of EVs (15k EVs and 100k EVs) were purified from adult F. gigantica culture media by ultracentrifugation. The morphology and size of the purified EVs were determined by transmission electron microscopy (TEM) and by the Zetasizer Nano ZSP high performance particle characterization system. With the aim of identifying diagnostic markers or potential vaccine candidates, proteins within the isolated 100k EVs were analyzed using mass spectrometry-based proteomics (LC-MS/MS). Mice were then vaccinated with excretory/secretory products (ESPs; depleted of EVs), 15k EVs, 100k EVs and recombinant F. gigantica heat shock protein 70 (rFg-HSP70) combined with alum adjuvant followed by challenge infection with F. gigantica metacercariae. Fluke recovery and antibody levels were used as measures of vaccine protection. RESULTS: TEM analysis and nanoparticle tracking analysis indicated the successful isolation of two subpopulations of EVs (15k EVs and 100k EVs) from adult F. gigantica culture supernatants using differential centrifugation. A total of 755 proteins were identified in the 100k EVs. Exosome biogenesis or vesicle trafficking proteins, ESCRT (endosomal sorting complex required for transport) pathway proteins and exosome markers, heat shock proteins and 14-3-3 proteins were identified in the 100k EVs. These results indicate that the isolated 100k EVs were exosome-like vesicles. The functions of the identified proteins may be associated with immune regulation, immune evasion and virulence. Mice immunized with F. gigantica ESPs, 15k EVs, 100k EVs and rFg-HSP70 exhibited a reduction in fluke burden of 67.90%, 60.38%, 37.73% and 56.6%, respectively, compared with the adjuvant control group. The vaccination of mice with F. gigantica 100k EVs, 15k EVs, ESP and rFg-HSP70 induced significant production of specific immunoglobulins in sera, namely IgG, IgG1 and IgG2a. CONCLUSION: The results of this study suggest that proteins within the exosome-like vesicles of F. gigantica have immunomodulatory, immune evasion and virulence functions. This knowledge may lead to new strategies for immunotherapy, vaccination and the diagnosis of fascioliasis.

[Ultrastructural observation on nymphal Armillifer sp. by scanning electron microscopy and phylogenetic analysis based on 18S rRNA].

OBJECTIVE: To observe the ultrastructure of nymphal Armillifer sp. isolated from Macaca fascicularis by using scanning electron microscope (SEM), and analyze the phylogenetic relationships based on 18S rRNA gene sequences. METHODS: The parasite samples stored in 70% alcohol were fixed by glutaraldehyde and osmium peroxide. Ultrastructural characters of those samples were observed under SEM. Amplification and sequencing of the 18S rRNA gene were performed following the extraction of total genome DNA. Sequence analysis was performed based on multiple alignment using ClustalX1.83, while phylogenetic analysis was made by Neighbor-Joining method using MEGA4.0. RESULTS: The nymphs were in cylindrical shape, the body slightly claviform tapering to posterior end. Abdominal annuli were gradually widened from anterior to posterior parts, the 12th-13th abdominal annuli of which were similar in width. The annuli ranged closer in the front half body, whereas in the latter part there were certain gaps between them. The circular-shaped mouth located in the middle of head ventrally. Folds were seen in inner margin of the mouth with a pair of curved hooks on both sides above it which practically disposed in a straight line. Two pairs of large sensory papillae were observed symmetrically over the last thoracic annulus of cephalothoraxs lying below the outer hook, and the first abdominal annulus was near the median ventral line. The number of abdominal annuli was 29, not including 2 incomplete terminal annuli. Rounded sensory papillae were fully distributed on the body surface, except the dorsal side of head and the ventral part of the terminal annulus. Agglomerate-like anus opening was observed at the end of ventral abdominal annuli and distinctly sub-terminal. These morphological features demonstrated that the nymphs were highly similar with that of Armillifer moniliformis Diesing, 1835. A fragment of 18SrRNA gene (1 836 bp) sequences was obtained by PCR combined with sequencing, and was registered to the GeneBank database with an accession number HM048870. The phylogenetic tree indicated that A. moniliformis, A.agkistrodon and A.armillatus were at the same clade with a bootstrap value at 95%, and A. moniliformis and A. agkistrodon were solo at a clade with a bootstrap value of 75%. CONCLUSION: The nymphs isolated from Macaca fascicularis are identified as A. moniliformis temporarily.

Prevalence of Lower Urinary Tract Symptoms in Children with Attention-Deficit/Hyperactivity Disorder: Comparison of Hospital and Population-Based Cohorts of 13,000 Patients.

Background: This study investigates the prevalence of lower urinary tract symptoms (LUTS) in school-age children with Attention-Deficit/Hyperactivity Disorder (ADHD) based on hospital-based and population-based cohorts. Methods: The hospital-based sample comprised 42 children with ADHD and 65 without ADHD aged 6−12 years. Voiding dysfunction was assessed by the Dysfunctional Voiding Scoring System (DVSS) questionnaire. We compared the baseline data, DVSS score, and uroflowmetry between the two groups. For the population-based cohort in the national insurance database, we included 6526 children aged 6−12 years, whose claims record included the diagnosis of ADHD, and another 6526 control subjects matched by gender and age. We compared the presence of LUTS diagnosis codes between the two groups. Results: Our results showed that, for the hospital-based cohort, the mean total DVSS score and the proportion of significant LUTS in children in the ADHD group were significantly higher than in subjects in the non-ADHD group. The DVSS subscales showed that the item "I cannot wait when I have to pee" item was significantly higher in the ADHD group (1.62 ± 1.17 vs. 0.90 ± 1.09, p = 0.002). For the population-based cohort, children with ADHD had a significantly higher likelihood of storage symptoms (5.53% vs. 2.91%, p < 0.001) and enuresis (3.28% vs. 1.95%, p < 0.001) compared with those of the no ADHD group. Conclusions: Children with ADHD have a higher prevalence of significant LUTS, especially storage symptoms and enuresis, than children without ADHD. The observed correlations between ADHD and LUTS provided the supporting evidence to evaluate the concomitant voiding dysfunction in children with ADHD.

Epidemiology of hypospadias and treatment trends in Taiwan: a nationwide study.

PURPOSE: We investigated the rate of diagnosis and treatment trends of hypospadias in Taiwan within the first 3 years of life. MATERIALS AND METHODS: We used a subset of the Taiwan National Health Insurance Research Database, which contains data on all inpatient and outpatient medical benefit claims, for the period 1997 through 2008 for a sample of 1 million individuals randomly drawn from the population of 25.68 million who held membership in the National Health Insurance program during any part of the calendar year 2005. We analyzed claims data for all subjects who were diagnosed with hypospadias through age 3 years. RESULTS: Among 52,705 newborns (individuals whose claims included live birth) 178 were diagnosed with hypospadias within 3 years of birth. Thus, mean incidence was 33.8 per 10,000 live male births. The hypospadias repair rate was 14.3 per 10,000 live male births. There was no significant tendency toward increase or decrease in rates of diagnoses or repairs, or proportion of severe hypospadias. There were significant associations between rates of hypospadias diagnoses and urbanization level of the community where the diagnosis was made. Approximately 54% of patients who underwent surgical repair did so after age 18 months. CONCLUSIONS: The nationwide incidence of hypospadias in children younger than 3 years in Taiwan is similar to that reported in previous studies. Delayed diagnosis and treatment of hypospadias are important clinical concerns that warrant further investigation.

[Detection and identification for Gnathostoma sp. in imported Monopterus albus].

OBJECTIVE: To inspect the third stage larvae of Gnathostoma in imported Monopterus albus, and identify its species. METHODS: Ten batches of M. albus imported to Shanghai were detected for nematode Gnathostoma from January 2010 to March 2011. Fifty-two M. albus imported from the Philippines (25), Indonesia (24) and Bangladesh (3) were sampled (3-10/batch), which were dissected, minced, and digested. The suspension was filtered with 10 mesh screen to take the deposit. The complete parasites were picked out under stereoscope followed by morphological identification. The rate and intensity of infection were calculated. Genomic DNA of Gnathostoma was extracted to amplify internal transcribed spacer region 2 (ITS-2) and cytochrome C oxidase subunit 1 (cox1) by PCR, the product of which was analyzed by electrophoresis and sequencing. The sequences were aligned with corresponding sequences in GenBank. RESULTS: The third stage larvae of Gnathostoma were detected in M. albus from Indonesia and Philippines with infection rate of 36.0% (9/25) and 50.0% (12/24) and average infectiosity of 7.8 (70/9) and 2.8 (34/12), respectively. No Gnathostoma was found in M. albus imported from Bangladesh. Under microscope, the larvae showed one cephalic bulb with 4 rings of hooklets on it, cross striations and small spines on the body surface. The front body spines were bigger and denser, while the rear spines were smaller and sparser. It had 1 cervical papilla and 4 cervical capsules. Morphological characteristics were similar to the third stage larvae of G. spinigerum. PCR results showed that the length of the ITS-2 and cox1 PCR products was 647 bp and 441 bp, respectively. Sequence alignment analysis showed that the two PCR products had 99%-100% consistency with G. spinigerum ITS-2 (GenBank Accession No. AB181155 and Z97175) and cox1 (GenBank Accession No. AY501388, AB180099, and AB551552). CONCLUSION: All the larvae detected in M. albus imported from the Philippines and Indonesia have been identified as G. spinigerum.

Estrogen receptor-associated receptor α and peroxisome proliferator-activated receptor γ in metabolism and disease (Review).

Estrogen receptor­associated receptor α (ERRα) is an orphan nuclear receptor that lacks corresponding ligands. ERRα recruits co­regulators to regulate gene transcription and plays an important role in human physiological functions. Peroxisome proliferator­activated receptor Î³ (PPARγ) is also a nuclear receptor that regulates the expression of target genes via a ligand­dependent mechanism, thereby participating in a series of physiological processes. Both ERRα and PPARγ are involved in the process of energy metabolism and tumorigenesis. In the present review, a concise overview of the important roles governed by ERRα and PPARγ in metabolism and their association with various disease are provided.

The role of lncRNA LSAT1 in the invasion and metastasis of non-small cell lung cancer under hypoxia.

BACKGROUND: To investigate the role of lncRNA LSAT1 in the hypoxia-induced invasion and metastasis of non-small cell lung cancer (NSCLC). METHODS: High-throughput microarrays were used to compare the lncRNA expression profile difference between normoxia-induced and hypoxia-induced NSCLC cell lines including A549, NCI-H1650 and NCI-H1299 in order to preliminarily screen key molecules related to hypoxia-induced invasion and metastasis of NSCLC. The different expression of lncRNA LSAT1 was measured in hypoxia-induced NSCLC cells (compared to normoxia-induced NSCLC cells) and 20 pairs of NSCLC tissues (compared to adjacent tissues) with RT-PCR. The expression of lncRNA LSAT1 in NSCLC cells A549 and NCI-H1299 was down-regulated by lentiviral transfection. Transwell migration and invasion assays and tail vein injection metastasis assay were employed for investigating the effect of lncRNA LSAT1 knockdown on the hypoxia-induced invasion and metastasis of NSCLC cells. RESULTS: RT-PCR showed that lncRNA LSAT1 was significantly increased in hypoxia-induced NSCLC cells A549, NCI-H1650 and NCI-H1299 and compared with adjacent tissues. The expression of lncRNA LSAT1 was also upregulated in NSCLC tissues. Transwell migration and invasion assays demonstrated that hypoxia could enhance the abilities of migration and invasion of NSCLC cells A549 and NCI-H1299. However, these two abilities were significantly inhibited after lncRNA LSAT1 knockdown. In addition, nude mice tail vein injection metastasis assay also verified that lncRNA LSAT1 knockdown inhibited liver metastasis and lung metastasis of NSCLC cell A549 in vivo. CONCLUSIONS: We found a series of hypoxia-related lncRNAs in NSCLC by means of high-throughput screening microarrays. Clinical specimen analysis and functional loss test confirmed that lncRNA LSAT1 is a key lncRNA for hypoxia-induced invasion and metastasis of NSCLC. In addition, STAT3 may be one of the target genes for lncRNA LSAT1 to play a regulatory role.