High-Coupled Magnetic-Levitation Hybrid Nanogenerator with Frequency Multiplication Effect for Wireless Water Level Alarm.

Hybrid nanogenerators (HNGs) represent a promising avenue for water energy harvesting, yet their commercial viability faces hurdles such as limited power output, poor coupling, and constrained operational lifespans. Here, a highly coupled triboelectric-electromagnetic magnetic-levitation hybrid nanogenerator (ML-HNG) is introduced that shows great potential for water energy harvesting. The ML-HNG fulfills the challenges of high power output, strong coupling, and long operational lifespans. During the contact-separation process of the triboelectric nanogenerator (TENG), the changing magnetic flux in the electromagnetic generator's coils generates a potential difference between the coils and Cu electrodes. The unique design of the ML-HNG employs a shared coil electrode configuration, which enhances the coupling without adding extra volume. This integration allows the ML-HNG to achieve multi-frequency vibrations and multiple output cycles per external longitudinal movement, a phenomenon known as the frequency multiplication effect. With an average power density of 1.69 W m-3 in water, the ML-HNG provides continuous power for a thermo-hygrometer and can quickly drive a wireless water level alarm system within a minute. This groundbreaking hybrid nanogenerator design holds significant promise for the efficient and consistent harvesting of low-frequency ocean wave energy, marking a substantial advancement in blue energy technology.

Untargeted metabolomics reveals potential plasma biomarkers for diagnosis of primary aldosteronism using liquid chromatography-mass spectrometry.

Metabolite profiling has the potential to comprehensively bridge phenotypes and complex heterogeneous physiological and pathological states. We performed a metabolomics study using parallel liquid chromatography-mass spectrometry (LC-MS) combined with multivariate data analysis to screen for biomarkers of primary aldosteronism (PA) from a cohort of 111 PA patients and 218 primary hypertension (PH) patients. Hydrophilic interaction chromatography and reversed-phase liquid chromatography separations were employed to obtain a global plasma metabolome of endogenous metabolites. The satisfactory classification between PA and PH patients was obtained using the MVDA model. A total of 35 differential metabolites were screened out and identified. A diagnostic biomarker panel was established using the least absolute shrinkage and selection operator (LASSO) binary logistic regression model and receiver operating characteristic analysis. Joint analysis with clinical indicators, including plasma supine aldosterone level, plasma orthostatic aldosterone level, body mass index, and blood potassium, revealed that the combination of metabolite biomarker panel and plasma supine aldosterone has the best clinical diagnostic efficacy.

Hsa_circ_0049396 inhibited oral squamous cell carcinoma progression by regulating the miR-663b/ENDOU axis.

BACKGROUND: Circular RNA (circRNAs) play an important role in oral squamous cell carcinoma (OSCC) progression and has been widely reported. In this study, we aimed to investigate the role of a novel circRNA, circ_0049396, and its underlying mechanism in OSCC. METHODS: The expression levels of circ_0049396, miR-663b, and theuridylate-specific endoribonuclease (ENDOU) were assessed by quantitative reverse transcription PCR (qRT-PCR). Cell proliferation and migration were evaluated using CCK-8 and Transwell assays, respectively. Western blotting was performed to measure the levels of the apoptosis-associated proteins (Bcl-2 and Bax). The functional role of circ _0049396 was further validated in a xenograft experiment in vivo. The interactions of miR-663b with circ_0049396/ENDOU were verified using the dual luciferase reporter, RNA pull-down, and RNA immunoprecipitation (RIP) assays. RESULTS: The expression of circ_0049396 and ENDOU was downregulated in OSCC tissues and cells, whereas miR-663b was upregulated. Circ_0049396 overexpression weakened OSCC cell proliferation and migration but enhanced their apoptosis. Circ_0049396 overexpression suppresses tumorigenesis in vivo. The circ_0049396/miR-663b/ENDOU regulatory network predicted through bioinformatic analysis was validated using RNA pull-down, luciferase reporter, and RIP experiments. MiR-663b mimic enhanced the migratory and proliferative abilities of OSCC cells, but suppressed apoptosis. Furthermore, circ_0049396 or ENDOU overexpression partially reversed the malignant behavior of miR-663b-overexpressing OSCC cells. CONCLUSIONS: Our study illustrated that circ_0049396 overexpression inhibited the malignant behavior of OSCC cells by regulating the miR-663b/ENDOU axis. Based on our findings, circ_0049396 can be used as a potential therapeutic agent for OSCC.

Elastic Self-Recovering Hybrid Nanogenerator for Water Wave Energy Harvesting and Marine Environmental Monitoring.

To achieve large-scale development of triboelectric nanogenerators (TENGs) for water wave energy harvesting and powering the colossal sensors widely distributed in the ocean, facile and scalable TENGs with high output are urgently required. Here, an elastic self-recovering hybrid nanogenerator (ES-HNG) is proposed for water wave energy harvesting and marine environmental monitoring. The elastic skeletal support of the ES-HNG is manufactured using three-dimensional (3D) printing technology, which is more conducive to the large-scale integration of the ES-HNG. Moreover, the combination of a TENG and an electromagnetic generator (EMG) optimizes the utilization of device space, leading to enhanced energy harvesting efficiency. Experimental results demonstrate that the TENG achieves a peak power output of 42.68 mW, and the EMG reaches a peak power output of 4.40 mW. Furthermore, various marine environment monitoring sensors, such as a self-powered wireless meteorological monitoring system, a wireless alarm system, and a water quality monitoring pen, have been successfully powered by the sophisticated ES-HNG. This work introduces an ES-HNG for water wave energy harvesting, which demonstrates potential in marine environment monitoring and offers a new solution for the sustainable development of the marine internet of things.

Cane Molasses Derived N-Doped Graphene Quantum Dots: Dynamic Quenching Synergistically Photoinduced Electron Transfer for the Instant Detection of Nitrofuran Antibiotics.

The development of a highly selective, simple, and rapid detection method for nitrofuran antibiotics (NFs) is of great significance for food safety, environmental protection, and human health. To meet these needs, in this work, cyan-color highly fluorescent N-doped graphene quantum dots (N-GQDs) were synthesized using cane molasses as the carbon source and ethylenediamine as the nitrogen source. The synthesized N-GQDs have an average particle size of 6 nm, a high fluorescence intensity with 9 times that of undoped GQDs, and a high quantum yield (24.4%) which is more than 6 times that of GQDs (3.9%). A fluorescence sensor based on N-GQDs for the detection of NFs was established. The sensor shows advantages of fast detection, high selectivity, and sensitivity. The limit of detection for furazolidone (FRZ) was 0.29 µM, the limit of quantification (LOQ) was 0.97 µM, and the detection range was 5-130 µM. The fluorescence quenching mechanism of the sensor was explored by fluorescence spectroscopy, UV-vis absorption spectroscopy, Stern-Volmer quenching constant, Zeta potential, UV-vis diffuse reflectance spectroscopy, and cyclic voltammetry. A fluorescence quenching mechanism of dynamic quenching synergized with photoinduced electron transfer was revealed. The developed sensor was also successfully applied for detecting FRZ in various real samples, and the results were satisfactory.

Dietary cholesterol consumption and incidence of type 2 diabetes mellitus: A dose-response meta-analysis of prospective cohort studies.

BACKGROUND AND AIM: The purpose of this meta-analysis was to evaluate the dose-response relationship between dietary cholesterol (DC) consumption and the incidence of type 2 diabetes mellitus (T2DM). METHODS AND RESULTS: Prospective studies with the endpoint of T2DM were included. The Random-effect model weighted by inverse variance was used. Meta-regression and subgroup analyses were conducted to explore the potential sources of heterogeneity by specified study characteristics. Restricted cubic splines regression models were used to estimate the dose-response relationship. 11 prospective studies comprising of 355 230 subjects were included. Compared to lowest DC consumption, highest DC consumption was associated with an increased risk of T2DM (RR 1.15, 95% CI 1.03 to 1.28, P = 0.012; chi-squared = 31.41, I-squared 58.6%, P heterogeneity = 0.003). Subgroup analyses have shown that this positive association was more evident in western countries than in eastern countries (RR 1.19, 95% CI 1.06 to 1.36 VS 1.34, 95% CI 0.84 to 1.29; P subgroup difference = 0.02). For 100 mg/d increment in DC intake, the pooled RR was 1.05, (95% CI 1.04 to 1.07, Plinearity = 0.000, Pnonlinearity = 0.02), 1.06 (95% CI 1.04 to 1.07, Plinearity=0.000), and 1.01 (95% CI 0.98 to 1.05, Plinearity = 0.525) for the incidence of T2DM, in western and eastern countries, respectively. CONCLUSIONS: Our study suggests that there is a positive dose-response association between DC consumption and the incidence of T2DM, especially in western countries. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42020216318.

Development and validation of a nomogram for individualized prediction of spontaneous extremely preterm birth at < 28 weeks in twin pregnancy: a retrospective cohort study.

PURPOSE: To develop a nomogram to predict spontaneous preterm birth at < 28 weeks in pregnant women with twin pregnancies. METHODS: We retrospectively studied the medical records of twin-pregnancy women with asymptomatic cervical dilation or cervical shortening between December 2015 to February 2022 in two hospitals. Data from one center was used to develop the model and data from the other was used to evaluate the model. RESULTS: A total of 270 twin pregnancies were enrolled in the study. We incorporated 4 items (cervical length, cervical dilation, C-reactive protein and the use of cerclage) to build the 28-week nomogram with satisfactory discrimination and calibration when applied to the validation sets. The C index for the 28-week nomogram in the development and external cohort was 0.88 (95% CI, 0.84-0.93) and 0.89 (95% CI, 0.80-0.98), respectively. The nomogram reached a sensitivity of 70.70%, specificity of 97.10%, positive predicted value of 95.61% and negative predicted value of 78.77%. Moreover, the decision curve analysis indicated that the nomogram showed positive clinical benefit. CONCLUSION: We developed and validated a nomogram with good performance in predicting individual risk of spontaneous preterm birth at < 28 in twin pregnancy.

Hypertensive status is associated with renoprotection by remote ischemic conditioning for acute myocardial infarction-a meta-regression and trial sequential analysis of randomized clinical trials.

The potential modifiable factors for remote ischemic conditioning (RIC) in reducing contrast-associated acute kidney injury (CA-AKI) in patients with acute myocardial infarction (AMI) have not been investigated. The aim of this meta-regression was to address these issues.We searched Pubmed, Embase and the Cochrane Library database for published randomized controlled trials (RCTs) with registration number CRD42020155532. Nine RCTs comprising of 1540 subjects were included in our meta-analysis. Compared with control group, RIC was associated with reduced incidence of CA-AKI [(9 studies, 1540 subjects, relative risk (RR) 0.51, 95% confidence intervals (CI) 0.35 to 0.76, p = 0.000, I2 = 52%, p for heterogeneity 0.04)] and major adverse cardiovascular events (MACE) (5 studies, 1078 subjects, RR 0.52, 95% CI 0.38 to 0.73, p = 0.000, I2 = 9%, p for heterogeneity 0.36) for AMI. In addition, both meta-regression and subgroup analyses have shown that RIC was more effective in the hypertensive patients in reducing CA-AKI for AMI (regression coefficient = -0.05, p = 0.021; for subgroup with more hypertensive patients: RR 0.36, 95% CI 0.25 to 0.52 vs the one with less hypertensive patients: RR 0.72, 95% CI (0.40 to 1.30, p for subgroup difference 0.008). Subsequent trial sequential analysis confirmed the effect of RIC in both CA-AKI and MACE. RIC is an effective strategy in reducing CA-AKI and MACE in patients with AMI, especially for patients with hypertension.

Immunogenicity of LY2963016 insulin glargine and Lantus® insulin glargine in Chinese patients with type 1 or type 2 diabetes mellitus.

AIMS: To evaluate the immunogenicity of LY2963016 insulin glargine (LY IGlar) versus originator insulin glargine (IGlar [Lantus®]) in Chinese patients with type 1 (T1DM) or type 2 diabetes mellitus (T2DM). MATERIALS AND METHODS: ABES and ABET were prospective, randomized, active control, open-label, phase III studies, which enrolled Chinese patients with T1DM (N = 272) and T2DM (N = 536), respectively. Using data from these trials, immunogenicity of LY IGlar and IGlar was evaluated by comparing the proportion of patients with detectable anti-insulin glargine antibodies and the median antibody levels (percent binding) between the treatment groups. The incidence of anti-insulin antibodies and treatment-emergent antibody response (TEAR) were compared using Fisher's exact test or Pearson's chi-squared test. Levels of anti-insulin antibodies were compared using the Wilcoxon rank-sum test. We also evaluated the relationship between antibody formation or TEAR and clinical outcomes using analysis of covariance, negative binomial regression, or partial correlations. RESULTS: There were no significant treatment differences in the incidence of detectable anti-insulin antibodies, median antibody levels or TEAR, overall or at Week 24 with last observation carried forward, and median antibody levels were low (<5%) after 24 weeks of treatment, in patients with T1DM or T2DM. Levels of anti-insulin antibodies and development of TEAR were not associated with efficacy (glycated haemoglobin, insulin dose [U/kg/d] and hypoglycaemia) or safety outcomes. CONCLUSIONS: The immunogenicity profiles of LY IGlar and IGlar are similar, with low levels of anti-insulin antibodies observed for both insulins. No association was observed between antibody levels or TEAR status and clinical outcomes.

Circ_0058063 contributes to oral squamous cell carcinoma development by sponging miR-145-5p and upregulating SERPINE1.

BACKGROUND: We aimed to demonstrate the effects circ_0058063 exerted on oral squamous cell carcinoma (OSCC) and its downstream mechanism associated with miR-145-5p and SERPINE1. METHODS: The relevant contents of miR-145-5p, circ 0058063, and SERPINE1 mRNAs in OSCC were evaluated using quantitative reverse transcription polymerase chain reaction. Functional experiments including CCK-8, Transwell, Western blot, and in vivo experiment were implemented to investigate the biological impacts on OSCC cells. Using dual-luciferase reporter, RIP, and RNA pull-down assays, the direct binding relationship between miR-145-5p, circ 0058063, and SERPINE1, SMAD3, CYR61, and IGF1R mRNAs was verified. RESULTS: In OSCC, Circ 0058063 was significantly overexpressed. Knockdown of circ_0058063 suppressed OSCC cell migration and proliferation, but enhanced cell apoptosis. Functionally and mechanistically, circ_0058063 could specifically bind with miR-145-5p and thus upregulated expression of downstream target SERPINE1, which together contributed to the progression of OSCC. CONCLUSION: Circ_0058063 could promote the malignant behavior of OSCC by upregulating SERPINE1 through sponging miR-145-5p.

Negative regulation of miR-1288-3p/KRT4 axis through a circular RNA in oral cancer.

Circular RNA (circRNA) has been widely reported to be involved in oral squamous cell carcinoma (OSCC), while the way in which hsa_circ_0096042 affects OSCC remains unclear. The hsa_circ_0096042, miR-1288-3p, and KRT4 expression in OSCC tissues and cell lines were detected by quantitative reverse-transcription polymerase chain reaction. Based on the overexpression of hsa_circ_0096042, miR-1288-3p, or KRT4, the viability and proliferation of OSCC cells were analyzed by cell counting kit-8 and colony formation assay, and the protein levels of Bax and Bcl-2 were detected by western blot, and the growth of cancer cells in vivo was analyzed by xenograft experiment. In addition, the database was used to predict the binding of hsa_circ_0096042, miR-1288-3p, and KRT4, and the interaction was confirmed by luciferase, RIP, and RNA pull-down assay. Hsa_circ_0096042 and KRT4 were abnormally downregulated and miR-1288-3p was upregulated in OSCC. Hsa_circ_0096042 overexpression restrained the proliferation and viability of OSCC cells, facilitated apoptosis, and inhibited the growth of cancer cells in vivo. Hsa_circ_0096042 bound to miR-1288-3p, whose upregulation promoted OSCC progression and eliminated the effects of overexpression of hsa_circ_0096042 on OSCC cells. KRT4 was the target gene for miR-1288-3p. Hsa_circ_0096042 plays an antitumor role in OSCC via miR-1288-3p/KRT4 axis.

A dilute-and-shoot liquid chromatography-tandem mass spectrometry method for urinary 18-hydroxycortisol quantification and its application in establishing reference intervals.

BACKGROUND: Eighteen-hydroxycortisol (18-OHF) is a potential biomarker for differential diagnosis of the two major primary aldosteronism subtypes, aldosterone-producing adenoma, and idiopathic hyperaldosteronism. METHODS: Urine samples were processed, and the 18-OHF in urine samples were successfully quantified by in-house established dilute-and-shoot liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. Separation was accomplished on a Sigma Ascentis Express C18 column with a gradient mixture of phase (A) 0.2% formic acid in water and phase (B) 0.2% formic acid in methanol at a flow rate of 0.4 ml/min. Mass spectrometric detection was performed in positive electrospray ionization mode via a mass spectrometer. RESULTS: The linearity of urinary 18-OHF ranged from 4.28 to 8.77 × 103 nmol/L, with a lower limit of quantification at 4.28 nmol/L. The intra- and inter-precision were both below 3%. The range of analytical recovery was 97.8%-109.2%. The validated dilute-and-shoot LC-MS/MS method was compared with the SPE LC-MS/MS method modified from the one reported in 2013. The results by Passing-Bablok regression analysis and Bland-Altman plotting demonstrated a good agreement between the two methods. The presented method was then applied to establish sex-specific reference intervals from 62 males and 62 females, respectively. The calculated 2.5%-97.5% reference intervals for 24-h urinary 18-OHF were 113-703 nmol/day for males and 71.2-450 nmol/day for females. CONCLUSION: The presented dilute-and-shoot LC-MS/MS method for 18-OHF quantification showed a good performance in the clinical application. Furthermore, the sex-specific reference intervals for 24-h urinary 18-OHF were first established and quite important for its application in primary aldosteronism subtyping.

Comparable efficacy and safety between LY2963016 insulin glargine and insulin glargine (Lantus®) in Chinese patients with type 1 diabetes: A phase III, randomized, controlled trial.

AIM: To compare the efficacy and safety of LY2963016 insulin glargine (LY IGlar) with the reference product (Lantus®) insulin glargine (IGlar) in Chinese patients with type 1 diabetes mellitus (T1DM). MATERIALS AND METHODS: This phase III, prospective, multicentre, open-label study enrolled patients with T1DM, age ≥18 years, with haemoglobin A1c (HbA1c) ≤11.0%, who were randomized to LY IGlar (n = 137) or IGlar (n = 135) in combination with premeal insulin lispro for 24 weeks. The treatment targets were to achieve HbA1c <7% and preprandial capillary blood glucose 79-126 mg/dl (4.4-7.0 mmol/L), avoiding hypoglycaemia. The primary efficacy endpoint was testing the non-inferiority of LY IGlar to IGlar by a margin of 0.4% using the mixed model repeated measure approach, as measured by changes in HbA1c levels from baseline to 24 weeks. Continuous laboratory measures were analysed using analysis of covariance. For categorical measures, Fisher's exact test was used. RESULTS: The least squares mean difference between treatments (LY IGlar - IGlar) in change from baseline was -0.12% (95% confidence interval -0.32%, 0.08%), meeting the non-inferiority criteria. There were no clinically meaningful differences (p > .05) in other efficacy outcomes, including proportions of patients achieving HbA1c <7.0% and HbA1c ≤6.5%, self-monitored blood glucose and insulin dose at week 24. Weight change, insulin antibodies and all adverse events including allergic reactions and hypoglycaemia, were also similar between the two treatment groups (all p > .05). CONCLUSIONS: LY IGlar and IGlar had equivalent efficacy in glycaemic control and similar safety profiles in Chinese patients with T1DM, when used in combination with mealtime insulin lispro.

A pragmatic study of mid-mixture insulin and basal insulin treatment in patients with type 2 diabetes uncontrolled with oral antihyperglycaemic medications: A lesson from real-world experience.

BACKGROUND: Chinese guidelines for the treatment of type 2 diabetes (T2D) recommend basal or premixed insulins as insulin starters after failed oral antihyperglycaemic medication (OAM). This pragmatic study compared effectiveness and safety of add-on basal insulin analog (BI) and mid-mixture insulin analog (MMI; 50:50 premixed insulin) as starter insulin regimens in Chinese patients with T2D in a real-world setting. MATERIALS AND METHODS: This was a multicentre, open-label, randomized, parallel, pragmatic trial. Patients receiving OAMs were randomized 1:1 to BI (n = 410) or MMI (n = 404) for 24 weeks. Insulin titration and OAM adjustment were determined by investigators following usual standard-of-care. The primary outcome was change in glycated haemoglobin (HbA1c) from baseline. RESULTS: Least-squares mean changes in HbA1c from baseline to week 24 were -2.00% and -2.15% for BI and MMI groups, respectively (P = .13). The MMI group demonstrated a greater reduction in concomitant OAM therapies used than BI group (53.8% vs. 35.3%, respectively; P < .001). Very limited daily insulin dose increments were observed from baseline to week 24 in both BI and MMI groups (2.5 U/day and 1.8 U/day, respectively). Although both insulin analogs were well-tolerated without severe hypoglycaemia, small weight gains were seen with both treatments. Higher total hypoglycaemia rates were noticed with the MMI group, while nocturnal hypoglycaemia events were comparable. CONCLUSIONS: In real-world settings, BI and MMI provided similar improvement in glucose control without conceding hypoglycaemia. The BI group received a greater number of OAMs in real-world settings. Limited insulin dose titration was observed, while more adjustments occurred with OAM usage.

Pre-procedural elevated cardiac troponin predict risk of long-term all-cause mortality after transcatheter aortic valve replacement: a meta-analysis of prospective studies.

Background: The purpose of this meta-analysis was to evaluate the relationship between elevated cardiac troponin pre-transcatheter aortic valve replacement (TAVR) and long-term all-cause mortality.Methods: Prospective studies with the endpoint of all-cause mortality were included. We primarily used the fixed-effect model weighted by inverse variance. Meta-regression and subgroup analyses were conducted to explore the potential sources of heterogeneity by specified study characteristics.Results: Seven prospective studies comprising of 3049 subjects were included in our meta-analysis. Pre-procedural elevated cardiac troponin was associated with increased risk of long-term mortality post TAVR [hazard ratio (HR) 2.25, 95% CI 1.83-2.78, p = 0.000, I2 = 30.3%, p for heterogeneity 0.197]. In addition, subgroup analyses have shown that the group with an younger age (<82 y) seemed to have a higher risk of all-cause mortality than the group with older age (≥82 y) [HR 4.08 (2.41 to 6.89) VS 2.01 (1.60 to 2.53), p = 0.016 for subgroup difference].Conclusions: Pre-procedural elevated cardiac troponin was associated with increased long-term all-cause mortality in patients undergoing TAVR.

BMC ecology image competition 2018: the winning images.

The sixth BMC Ecology Image Competition received more than 145 photographs from talented ecologists around the world, showcasing the amazing biodiversity, natural beauty and biological interactions found in nature. In this editorial, we showcase the winning images, as selected by our guest judge, Professor Zhigang Jiang from the Institute of Zoology of the Chinese Academy of Sciences, with help from the journal's editorial board. Enjoy!

Efficacy and safety of tadalafil 5 mg once-daily in Asian men with both lower urinary tract symptoms associated with benign prostatic hyperplasia and erectile dysfunction: A phase 3, randomized, double-blind, parallel, placebo- and tamsulosin-controlled study.

OBJECTIVE: To evaluate the efficacy and safety of tadalafil in Asian men with both lower urinary tract symptoms associated with benign prostatic hyperplasia and erectile dysfunction. METHODS: The present phase 3, randomized, double-blind, parallel, placebo- and tamsulosin-controlled study was carried out at 40 study centers in the Asia-Pacific region (mainland China, Taiwan and Korea; NCT01937871). Participants were randomized to receive a placebo (n = 361), tadalafil 5 mg (n = 362) or tamsulosin 0.2 mg (n = 185) in a 2:2:1 ratio for 12 weeks. RESULTS: A total of 909 Asian men were randomized into three groups. After 12 weeks of treatment, a statistically significant improvement in least squares mean change from baseline in total International Prostate Symptom Score was observed in the tadalafil versus the placebo group (-5.49 vs -4.08, respectively; P < 0.001). A statistically significant improvement in the change from baseline for the International Index of Erectile Function-Erectile Function domain score, was observed in tadalafil compared with the placebo at 12 weeks (5.24 vs 1.88, respectively; P < 0.001). A significant improvement was observed in the change from baseline in the percentage of "Yes" responses to Sexual Encounter Profile questions 2 and 3 in the tadalafil versus placebo group at 12 weeks (23.87% vs 10.90%; P < 0.001 and 36.62% vs 15.96%; P < 0.001, respectively). Safety results were consistent with the known tadalafil safety profile. CONCLUSIONS: Tadalafil is efficacious and well tolerated in the treatment of Asian men with both lower urinary tract symptoms associated with benign prostatic hyperplasia and erectile dysfunction.

Secretory expression and scale-up production of recombinant human thyroid peroxidase via baculovirus/insect cell system in a wave-type bioreactor.

The human thyroid peroxidase (hTPO) is an essential enzyme for thyroid hormone biosynthesis and is expressed in thyroid cells. It is an autoantigen against which antibodies are found in the sera of patients with a number of autoimmune thyroid disorders. Overexpression of hTPO has been achieved using the baculovirus expression vector system (BEVS). However, it is produced largely in an aggregated form in the cell lysate fraction, which increases the complexity of protein extraction. In this study, to achieve improved secretory expression of hTPO via BEVS, a truncated recombinant hTPO protein (hTPOt) was engineered by replacing its original signal peptide (SP) in the N-terminal with five heterologous SPs. Our data showed that the SP from the peptidyl-glycine alpha-amidating monooxygenase (PAM), referred to as SPPAM, significantly promoted the secretion of SPPAM-fused hTPOt (p-hTPOt) in High Five cells. Subsequently, we established an optimized scale-up production procedure for p-hTPOt in a 5-L wave-type bioreactor. The secretory p-hTPOt was purified by immobilized metal-chelating affinity chromatography and ion-exchange chromatography, achieving a protein purity of >95%. Finally, the purified p-hTPOt showed high sensitivity and specificity in reactions with positive or negative human serum samples via the double-antigen sandwich method, suggesting potential applications in hTPO-based research and product development.

Intracellular translocation and localization of Edwardsiella tarda type III secretion system effector EseG in host cells.

Edwardsiella tarda, an important fish pathogenic bacterium, could utilize type III secretion system (T3SS) to transfer multiple effector proteins into host cells during infection. EseG was identified to be an E. tarda T3SS effector, which could be injected by T3SS into non-phagocytic cells. Since E. tarda is a facultative intracellular pathogen that resides and replicates in macrophage, it is interesting to expand our knowledge about EseG translocation and localization within phagocytic cells. Here utilizing murine macrophage cell line J774A.1 as the cell model, we demonstrated that EseG could be transported into J774A.1 via T3SS only after E. tarda was internalized into macrophage cells, indicating that extracellular E. tarda could not inject EseG into host cells. Subcellular fractionation analysis gave the evidence that EseG was specifically localized in the membrane fraction of infected host cells. Furthermore, immunofluorescence detection indicated that EseG specifically targeted the E. tarda-containing vacuoles (ECVs) within macrophage cells. Finally the unique features for EseG were also confirmed in non-phagocytic cells. In summarize, this work illuminates internalization-depending translocation and ECV-targeting localization of E. tarda T3SS effector in both non-phagocytic and phagocytic cells, which might be important to interpret the interaction of EseG with host cells upon infection.