Pain perception, aqueous humour cytokines, and miosis response following first and second eye femtosecond laser-assisted cataract surgery: A randomised, prospective, intraindividual study.

BACKGROUND: To compare pain perception, pupil behaviours, and cytokine levels during first-eye and second-eye femtosecond laser-assisted cataract surgery (FLACS) and determine which is better regarding a short or long interval for bilateral FLACS. METHODS: Notably, 96 eyes of 48 patients with bilateral cataracts underwent the first surgeries in the left or right eye, according to a random sequence. They were further randomised into 2- and 6-week subgroups based on surgery intervals. Pupil size was measured from captured images, and pain perception was assessed using a visual analog scale (VAS). Aqueous humour prostaglandin E2 (PGE2), monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-6, and IL-8 levels were also quantitatively analysed. All patients were followed for 1 week to evaluate changes in endothelial cell density (ECD), central corneal thickness (CCT), and macular central subfield thickness (CST). RESULTS: Ocular pain was significantly higher in patients who underwent second-eye FLACS. First tear break-up time was negatively correlated with VAS score. MCP-1 levels were significantly higher in patients who underwent second-eye FLACS, and VAS scores were positively correlated with MCP-1 levels across all patients. There were no differences between sequential FLACS in miosis, PGE2, IL-6, IL-8 levels and changes in postoperative ECD, CCT, and CST. Patients who underwent second-eye FLACS after 6 weeks showed more CCT, CST, and MCP-1 changes than baseline. CONCLUSION: Second-eye FLACS causes more pain and upregulated MCP-1. There was no difference between sequential FLACS in PGE2 levels, miosis, and postoperative inflammation. Furthermore, first-eye FLACS triggered a sympathetic irritation, particularly after a 6-week interval.

Sustainable reprocessing of lithium iron phosphate batteries: A recovery approach using liquid-phase method at reduced temperature.

Lithium iron phosphate batteries, known for their durability, safety, and cost-efficiency, have become essential in new energy applications. However, their widespread use has highlighted the urgency of battery recycling. Inadequate management could lead to resource waste and environmental harm. Traditional recycling methods, like hydrometallurgy and pyrometallurgy, are complex and energy-intensive, resulting in high costs. To address these challenges, this study introduces a novel low-temperature liquid-phase method for regenerating lithium iron phosphate positive electrode materials. By using N2H4·H2O as a reducing agent, missing Li+ ions are replenished, and anti-site defects are reduced through annealing. This process restores nearly all missing Li+ ions at 80 °C/6h. After high-temperature sintering at 700 °C/2h, the regenerated LiFePO4 matches commercial LiFePO4 in terms of anti-site defects and exhibits excellent performance with a 97 % capacity retention rate after 100 cycles at 1C. Compared to high-temperature techniques, this low-temperature liquid-phase method is simpler, safer, and more energy-efficient, offering a blueprint for reclaiming discarded LiFePO4 and similar materials.

Bismuth-Containing Quadruple Therapy for Helicobacter pylori Eradication: A Randomized Clinical Trial of 10 and 14 Days.

BACKGROUND: Bismuth-containing quadruple therapy is the first-line treatment for eradicating Helicobacter pylori (H. pylori). The optimal duration for H. pylori eradication using bismuth-containing quadruple therapy remains controversial. Therefore, we aimed to compare the clinical effects of the 10- and 14-day bismuth-containing quadruple treatment regimen to eradicate H. pylori. METHODS: Treatment-naïve patients with H. pylori infection (n = 1300) were enrolled in this multicenter randomized controlled study across five hospitals in China. They were randomized into 10- or 14-day treatment groups to receive bismuth-containing quadruple therapy as follows: vonoprazan 20 mg twice daily; bismuth 220 mg twice daily; amoxicillin 1000 mg twice daily; and either clarithromycin 500 mg twice daily or tetracycline 500 mg four times daily. At least 6 weeks after treatment, we performed a 13C-urea breath test to evaluate H. pylori eradication. RESULTS: The per-protocol eradication rates were 93.22% (564/605) and 93.74% (569/607) (p < 0.001) and the intention-to-treat eradication rates were 88.62% (576/650) and 89.38% (581/650) (p = 0.007) for the 10- and 14-day regimens, respectively. Incidence of adverse effects was lower in patients who received 10- vs. 14 days of treatment (22.59% vs. 28.50%, p = 0.016). We observed no significant differences in the compliance to treatment or the discontinuation of therapy because of severe adverse effects between the groups. CONCLUSION: Compared with the 14-day bismuth-containing quadruple regimens, the 10-day regimen demonstrated a non-inferior efficacy and lower incidence of adverse effects. Therefore, the 10-day regimen is safe and tolerated and could be recommended for H. pylori eradication (NCT05049902).

A novel intraocular pressure predicting method based on hyperelastic mechanical model of cornea.

Measuring intraocular pressure (IOP) is crucial and remains challenging in diagnosing glaucoma, as it is associated with cornea deformation during inflation. In this study, a three-dimensional analytical model based on hyperelastic constitutive relationship to predict correlation between cornea vertex displacement and the IOP is proposed. The analytical model is validated by rigorous experiments. Rabbit corneas were selected for this study and their mechanical properties were obtained using uniaxial tensile tests. To mimic the environment in which the cornea exists, an artificial anterior chamber equipped with water-injection pipelines was constructed to study the relationship between the corneal vertex displacement with IOP value in practical situation. The experimental results of rabbits corneas prove that the IOP can be deduced based on the measured corneal vertex displacement by the analytical model. Furthermore, subtle difference occurs when comparing the calculated human IOPs with those measured by medical equipment, demonstrating that the proposed method is suitable for monitoring the IOP of human. This novel IOP predicting method provides new inspiration for the design of eyepieces, as well as the preoperative preparation for laser surgery and evaluation of corneal damage.

Design and Processing of Gas Turbine Blades Based on Additive Manufacturing Technology.

Aiming at the problems of the complex shape, difficult three-dimensional (3D) digital modeling and high manufacturing quality requirements of gas turbine blades (GTB), a method of fitting the blade profile line based on a cubic uniform B-spline interpolation function was proposed. Firstly, surface modeling technology was used to complete the fitting of the blade profile of the GTB, and the 3D model of the GTB was synthesized. Secondly, the processing parameters of the additive manufacturing were set, and the GTB model was printed by fused deposition technology. Then, the rapid investment casting was completed with the printed model as a wax model to obtain the GTB casting. Finally, the blade casting was post-processed and measured, and it was found to meet the requirements of machining accuracy and surface quality.

Facile Synthesis of Self-Targeted Zn2+ -Gallic acid Nanoflowers for Specific Adhesion and Elimination of Gram-Positive Bacteria.

Transition metal ions are served as disinfectant thousand years ago. However, the in vivo antibacterial application of metal ions is strongly restricted due to its high affinity with proteins and lack of appropriate bacterial targeting method. Herein, for the first time, Zn2+ -gallic acid nanoflowers (ZGNFs) are synthesized by a facile one-pot method without additional stabilizing agents. ZGNFs are stable in aqueous solution while can be easily decomposed in acidic environments. Besides, ZGNFs can specifically adhere onto Gram-positive bacteria, which is mediated by the interaction of quinone from ZGNFs and amino groups from teichoic acid of Gram-positive bacteria. ZGNFs exhibit high bactericidal effect toward various Gram-positive bacteria in multiple environments, which can be ascribed to the in situ Zn2+ release on bacterial surface. Transcriptome studies reveal that ZGNFs can disorder basic metabolic processes of Methicillin-resistant Staphylococcus aureus (MRSA). Moreover, in a MRSA-induced keratitis model, ZGNFs exhibit long-term retention in the infected corneal site and prominent MRSA elimination efficacy due to the self-targeting ability. This research not only reports an innovative method to prepare metal-polyphenol nanoparticles, but also provides a novel nanoplatform for targeted delivery of Zn2+ in combating Gram-positive bacterial infections.

Quantum simulation of Hawking radiation and curved spacetime with a superconducting on-chip black hole.

Hawking radiation is one of the quantum features of a black hole that can be understood as a quantum tunneling across the event horizon of the black hole, but it is quite difficult to directly observe the Hawking radiation of an astrophysical black hole. Here, we report a fermionic lattice-model-type realization of an analogue black hole by using a chain of 10 superconducting transmon qubits with interactions mediated by 9 transmon-type tunable couplers. The quantum walks of quasi-particle in the curved spacetime reflect the gravitational effect near the black hole, resulting in the behaviour of stimulated Hawking radiation, which is verified by the state tomography measurement of all 7 qubits outside the horizon. In addition, the dynamics of entanglement in the curved spacetime is directly measured. Our results would stimulate more interests to explore the related features of black holes using the programmable superconducting processor with tunable couplers.

Association between one-year exposure to air pollution and the prevalence of pulmonary nodules in China.

PM2.5is a well-known airborne hazard to cause various diseases. Evidence suggests that air pollution exposure contributes to the occurrence of pulmonary nodules. Pulmonary nodules detected on the computed tomography scans can be malignant or progress to malignant during follow-up. But the evidence of the association between PM2.5exposure and pulmonary nodules was limited. To examine potential associations of exposures to PM2.5and its major chemical constituents with the prevalence of pulmonary nodules. A total of 16 865 participants were investigated from eight physical examination centers in China from 2014 to 2017. The daily concentrations of PM2.5and its five components were estimated by high-resolution and high-quality spatiotemporal datasets of ground-level air pollutants in China. The logistic regression and the quantile-based g-computation models were used to assess the single and mixture impact of air pollutant PM2.5and its components on the risk of pulmonary nodules, respectively. Each 1 mg m-3increase in PM2.5(OR 1.011 (95% CI: 1.007-1.014)) was positively associated with pulmonary nodules. Among five PM2.5components, in single-pollutant effect models, every 1µg m-3increase in organic matter (OM), black carbon (BC), and NO3-elevated the risk of pulmonary nodule prevalence by 1.040 (95% CI: 1.025-1.055), 1.314 (95% CI: 1.209-1.407) and 1.021 (95% CI: 1.007-1.035) fold, respectively. In mixture-pollutant effect models, the joint effect of every quintile increase in PM2.5components was 1.076 (95% CI: 1.023-1.133) fold. Notably, NO3-BC and OM contributed higher risks of pulmonary nodules than other PM2.5components. And the NO3-particles were identified to have the highest contribution. The impacts of PM2.5components on pulmonary nodules were consistent across gender and age.These findings provide important evidence for the positive correlation between exposure to PM2.5and pulmonary nodules in China and identify that NO3-particles have the highest contribution to the risk.

New insights into the substrate inhibition of human 17ß-hydroxysteroid dehydrogenase type 1.

Human type 1 17ß-hydroxysteroid dehydrogenase (17ß-HSD1),a member of the short-chain dehydrogenase/reductase family, catalyzes the last step in the bioactivation of the most potent estrogen estradiol with high specificity and is thus involved in estrogen-dependent diseases. As an oxidoreductase, 17ß-HSD1 can utilize both triphosphate and diphosphate cofactors in reaction at the molecular level, but more specific with triphosphate cofactor. The NADPH is much higher than NADP+ in living cells leading to preliminary reduction action. The enzyme also showed substrate-induced inhibition unprecedented in other members of 17ß-HSDs. Our previous study elucidated the structural mechanism of substrate inhibition is due to the reversely bound estrone (E1) in the substrate-binding pocket of the enzyme resulting in a dead-end complex. However, the effect of the cofactor preference on the substrate inhibition of the enzyme is not yet clear. In the present study, we solved the ternary crystal structures of 17ß-HSD1 in complex with E1 and cofactor analog NAD+ . Combined with molecular dynamics simulation using the enzyme with NADH/NADPH and different oriented E1 (normally oriented, E1N; reversely oriented, E1R), such ternary structure provides a complete picture of enzyme-substrate-cofactor interactions. The results reveal that different cofactors and substrate binding mode affect the allosteric effect between the two subunits of the enzyme. And the results from MD simulations confirmed that His221 plays a key role in the formation of dead-end complex in NADPH complex, and the absence of stable interaction between His221 and E1R in the NADH complex should be the main reason for its lack of substrate inhibition.

Comparative visual outcomes of EDOF intraocular lens with FLACS vs conventional phacoemulsification.

PURPOSE: To investigate the visual quality after femtosecond laser-assisted cataract surgery (FLACS) and conventional phacoemulsification surgery (CPS) with the implantation of an extended depth-of-focus (EDOF) intraocular lens (IOL) TECNIS Symfony. SETTING: Eye Center, the Second Affiliated Hospital of Zhejiang University, Zhejiang, China. DESIGN: Prospective cohort study. METHODS: Patients were given the option to choose FLACS or CPS and were implanted with a TECNIS Symfony. Main outcome measures were postoperative examinations that included defocus curves, contrast sensitivity (CS), optic path difference aberrometry scan, anterior segment photography, and questionnaires. RESULTS: 261 patients (261 eyes) were enrolled. The circularity index of FLACS was more precise than that of CPS ( P = .001). FLACS demonstrated a significantly lower IOL decentration ( P = .011) and IOL tilt ( P = .009). FLACS presented a significantly lower total aberration ( P < .001), higher-order aberrations (HOAs) ( P = .001), coma ( P = .001), and spherical aberration ( P < .001). With IOL decentration of more than 0.40 mm, total internal aberration ( P = .023) and HOAs ( P = .045) were significantly deteriorated. As for defocus curve, FLACS was better at -1.00 diopter ( P < .01). The FLACS group achieved higher CS at 6 to 18 cycles per degree under glare condition ( P < .05). With regard to photic phenomena, the FLACS group received better outcomes ( P < .05). CONCLUSIONS: With the implantation of an EDOF IOL, FLACS could precisely control the shape and size of the capsulotomy and induce a significantly better-centered IOL, leading to higher visual performance compared with CPS.

Fabrication and properties of interweaved poly(ether ether ketone) composite scaffolds.

This paper interweaved scaffolds with poly(ether ether ketone) (PEEK) and poly(lactic acid)/Walnut shell/hydroxypatite (PLA/WS/HA) composites by using fused filament fabrication technology, although there was a huge difference in thermal property term between PLA and PEEK. In order to keep mechanical properties of PEEK scaffold and remedy the stress loss produced by pores, PLA/WS/HA composites were used to fill the pores with gradient form outside-in (0.4-0.8 mm, 0.6-1.0 mm, 0.8-1.2 mm and 1.6-2.0 mm). The thermal stability, tensile and compression properties, tensile fracture surface morphology, cytotoxicity and in vivo experiment were investigated. The results showed: the scaffolds were intact without any flashes and surface destruction, and kept a well thermal stability. Compared with the PEEK porous scaffolds, the tensile fracture stress and strain, compression yield stress and strain of interweaved scaffolds were dramatically enhanced by 24.1%, 438%, 359.1% and 921.2%, respectively, and they climbed to the climax at 8 wt% of WS. In vivo experiment showed that the degradation of PLA/WS/HA composites synchronized with the adhesion, proliferation and ingrowth of bone cells, keeping the stable biomechanical properties of interweaved scaffolds. Those experiments showed that interweaved PEEK-PLA/WS/HA scaffolds had the potential to be used as bone implant in tissue engineering.

Distributions and trends of the global burden of COPD attributable to risk factors by SDI, age, and sex from 1990 to 2019: a systematic analysis of GBD 2019 data.

BACKGROUND: Global distributions and trends of the risk-attributable burdens of chronic obstructive pulmonary disease (COPD) have rarely been systematically explored. To guide the formulation of targeted and accurate strategies for the management of COPD, we analyzed COPD burdens attributable to known risk factors. METHODS: Using detailed COPD data from the Global Burden of Disease study 2019, we analyzed disability-adjusted life years (DALYs), years lived with disability (YLDs), years of life lost (YLLs), and deaths attributable to each risk factor from 1990 to 2019. Additionally, we calculated estimated annual percentage changes (EAPCs) during the study period. The population attributable fraction (PAF) and summary exposure value (SEV) of each risk factor are also presented. RESULTS: From 1990 to 2019, the age-standardized DALY and death rates of COPD attributable to smoking and household air pollution, occupational particles, secondhand smoke, and low temperature presented consistently declining trends in almost all socio-demographic index (SDI) regions. However, the decline in YLD was not as dramatic as that of the death rate. In contrast, the COPD burden attributable to ambient particulate matter, ozone, and high temperature exposure showed undesirable increasing trends in the low- and low-middle-SDI regions. In addition, the age-standardized DALY and death rates attributable to each risk factor except household air pollution and low temperature were the highest in the low-middle-SDI region. In 2019, the COPD burden attributable to smoking ambient particulate matter, ozone, occupational particles, low and high temperature was obviously greater in males than in females. Meanwhile, the most important risk factors for female varied across regions (low- and low-middle-SDI regions: household air pollution; middle-SDI region: ambient particles; high-middle- and high-SDI region: smoking). CONCLUSIONS: Increasing trends of COPD burden attributable to ambient particulate matter, ozone, and high temperature exposure in the low-middle- and low-SDI regions call for an urgent need to implement specific and effective measures. Moreover, considering the gender differences in COPD burdens attributable to some risk factors such as ambient particulate matter and ozone with similar SEV, further research on biological differences between sexes in COPD and relevant policy-making of disease prevention are required.

NUAK2 silencing inhibits the proliferation, migration and epithelial­to­mesenchymal transition of cervical cancer cells via upregulating CYFIP2.

NUAK family kinase 2 (NUAK2) has been reported to be involved in various cancer cell processes, including proliferation, apoptosis and invasion, by targeting multiple genes. However, to the best of our knowledge, its biological function in cervical cancer (CC) has not yet been elucidated. Therefore, the present study aimed to measure the expression of NUAK2 and to evaluate its functions in CC. The expression levels of NUAK2 and cytoplasmic FMRP­interacting protein 2 (CYFIP2) were detected in CC tissues and cell lines. In addition, the effects of NUAK2 and CYFIP2 knockdown on CC cell proliferation, migration, invasion and epithelial­to­mesenchymal transition (EMT) were evaluated in vitro using Cell Counting Kit­8, immunofluorescence, wound healing assay, Transwell assay and western blotting, respectively. Furthermore, co­immunoprecipitation was performed to determine the interaction between NUAK2 and CYFIP2. The results revealed that the expression levels of NUAK2 were upregulated in CC tissues and cells, whereas CYFIP2 expression was reduced. In addition, knockdown of NUAK2 reduced cell proliferation, migration, invasion and EMT. Notably, NUAK2 was found to bind directly to CYFIP2. Furthermore, CYFIP2 inhibition reversed the effects of NUAK2 on CC cells. In summary, NUAK2 may regulate CYFIP2 expression to promote CC cell proliferation, migration, invasion and EMT.

Paternal USP26 mutations raise Klinefelter syndrome risk in the offspring of mice and humans.

Current understanding holds that Klinefelter syndrome (KS) is not inherited, but arises randomly during meiosis. Whether there is any genetic basis for the origin of KS is unknown. Here, guided by our identification of some USP26 variations apparently associated with KS, we found that knockout of Usp26 in male mice resulted in the production of 41, XXY offspring. USP26 protein is localized at the XY body, and the disruption of Usp26 causes incomplete sex chromosome pairing by destabilizing TEX11. The unpaired sex chromosomes then result in XY aneuploid spermatozoa. Consistent with our mouse results, a clinical study shows that some USP26 variations increase the proportion of XY aneuploid spermatozoa in fertile men, and we identified two families with KS offspring wherein the father of the KS patient harbored a USP26-mutated haplotype, further supporting that paternal USP26 mutation can cause KS offspring production. Thus, some KS should originate from XY spermatozoa, and paternal USP26 mutations increase the risk of producing KS offspring.

Human Ccr4 and Caf1 Deadenylases Regulate Proliferation and Tumorigenicity of Human Gastric Cancer Cells via Modulating Cell Cycle Progression.

Cancer cells generally have reprogrammed gene expression profiles to meet the requirements of survival, continuous division, and metastasis. An interesting question is whether the cancer cells will be affected by interfering their global RNA metabolism. In this research, we found that human Ccr4a/b (hCcr4a/b) and Caf1a/b (hCaf1a/b) deadenylases, the catalytic components of the Ccr4-Not complex, were dysregulated in several types of cancers including stomach adenocarcinoma. The impacts of the four deadenylases on cancer cell growth were studied by the establishment of four stable MKN28 cell lines with the knockdown of hCcr4a/b or hCaf1a/b or transient knockdown in several cell lines. Depletion of hCcr4a/b or hCaf1a/b significantly inhibited cell proliferation and tumorigenicity. Mechanistic studies indicated that the cells were arrested at the G2/M phase by knocking down hCaf1a, while arrested at the G0/G1 phase by depleting hCaf1b or hCcr4a/b. The four enzymes did not affect the levels of CDKs and cyclins but modulated the levels of CDK-cyclin inhibitors. We identified that hCcr4a/b, but not hCaf1a/b, targeted the p21 mRNA in the MKN28 cells. Furthermore, depletion of any one of the four deadenylases dramatically impaired processing-body formation in the MKN28 and HEK-293T cells. Our results highlight that perturbating global RNA metabolism may severely affect cancer cell proliferation, which provides a potential novel strategy for cancer treatment.

Visual outcome and optical quality after implantation of zonal refractive multifocal and extended-range-of-vision IOLs: a prospective comparison.

PURPOSE: To compare the visual outcomes and optical quality of 2 presbyopia-correcting intraocular lenses (IOLs) with those of a monofocal IOL. SETTINGS: Eye Center, the Second Affiliated Hospital of Zhejiang University, School of Medicine, Hangzhou, Zhejiang, China. DESIGN: Prospective cohort study. METHODS: The study included patients who had cataract surgery and were implanted with a Tecnis Symfony Extended Range of Vision (EROV) IOL (ZXR00), a zonal refractive multifocal IOL (Lentis Comfort LS-313 MF15), or a monofocal IOL (Lentis L-313). Postoperative examinations took place at 1 week, 1 month, and 3 months and included visual acuity at far, intermediate, and near distances, defocus curves, contrast sensitivity, wavefront aberrations, and modulation transfer function (MTF). Patients completed the Visual Function Index questionnaire (VF-14), the Quality of Vision questionnaire (QoV), and a visual quality self-evaluation. RESULTS: One hundred thirteen patients were enrolled. The EROV and multifocal IOLs achieved a significantly better range of intermediate vergences (P < .05), better distance-corrected intermediate visual acuity (P ≤ .001), higher VF-14 (P < .05) and visual quality self-evaluation scores (P < .05) than the monofocal IOL, but there were no significant differences between the 2 presbyopia-correcting IOLs. The EROV provided lower total wavefront aberrations and better MTF than the multifocal and the monofocal IOLs (P < .05) but demonstrated a worse QoV score (P < .05), especially for severity of halo (P < .01) and starburst (P < .05) symptoms. CONCLUSIONS: Both the Tecnis Symfony ZXR00 and the Lentis Comfort LS-313 MF15 offered excellent visual restoration and stable distance and intermediate visual acuity, good subjective visual function, and good contrast sensitivity. The EROV IOL provided better objective optical quality and more prominent dysphotopsia symptoms than the multifocal IOL.

Comparison of the Clinical Outcomes between Echelette Extended Range of Vision and Diffractive Bifocal Intraocular Lenses.

PURPOSE: To compare the clinical outcomes of echelette extended range of vision (ERV) and diffractive bifocal intraocular lenses (IOLs). METHODS: This is a prospective, consecutive, nonrandomized clinical trial. Seventy-three eligible patients (109 eyes) received the implantation of echelette ERV IOL (Tecnis Symfony ZXR00) or diffractive bifocal IOL (Tecnis ZMB00). 1 week, 1 month, and 3 months after surgery, visual acuities at different distances were examined. At 3 months, defocus curves, contrast sensitivities (CSs) with and without glare, optic path difference (OPD) scans, and questionnaires were evaluated. Regression analyses were applied to discover influence factors on postoperative vision. RESULTS: ZXR00 showed better distance (P < 0.05) and intermediate (P < 0.001) visual acuities, while ZMB00 was better at distance-corrected near visual acuity (P < 0.001). Multivariate analyses indicated that worse intermediate (P < 0.001) and near vision (P=0.013) of ZMB00 might occur in patients with longer axial length. ZXR00 demonstrated smoother defocus curve and higher CSs. Superior modulation transfer function (MTF) and higher Strehl ratio (P < 0.05) were shown in ZXR00. In questionnaire evaluation, ZXR00 received better outcomes in self-reported vision, Visual Function-14 (VF-14) questionnaire, Quality of Vision (QoV) questionnaire, satisfaction, and recommendation grades. Spectacle dependence did not differ between ZXR00 and ZMB00 statistically. CONCLUSION: ZXR00 proved to be remarkable in distance and intermediate vision, defocus curve smoothness, CSs, and visual comfort, while ZMB00 achieved better near vision. ZXR00 may attain better near vision if postoperative SE remains slightly negative. Patients with relatively longer axial length might receive less favorable intermediate and near vision after ZMB00 implantation. This trial is registered with ChiCTR-ONC-17011119.

Assembled Step Emulsification Device for Multiplex Droplet Digital Polymerase Chain Reaction.

Digital PCR is a powerful method for absolute nucleic acid quantification with unprecedented accuracy and precision. To promote the wider use and application of digital PCR, several major challenges still exist, including reduction of cost, integration of the instrumental platform, and simplification of operations. This paper describes a reusable microfluidic device that generates nanoliter droplet arrays based on step emulsification for the on-chip multiplex digital PCR of eight samples simultaneously. The device contains two glass plates that can be quickly assembled with prefilled mineral oil. Droplets are simply generated through the arrays of step emulsification nozzles driven by a single pressure controller and are self-assembled into monolayer droplet arrays in U-shaped chambers. The use of mineral oil eliminates bubble generation; thus, no overpressure is required during thermocycling. Moreover, the device can be reused many times after disassembly and a brief cleaning procedure, which significantly reduces the cost of the device per dPCR assays. The device was able to detect template DNA at concentrations as low as 10 copies/µL with a dynamic range of approximately 4 logs. We applied this device in the quantitative assessment of HER2 copy number variation, which is important for targeted therapy and prognosis of breast cancer. The performance was validated by 16 clinical samples, obtaining similar results to commercial digital PCR. We envision that this low-cost, reusable, and user-friendly device can be broadly used in various applications.

Detection and characterization of ESBL-producing Escherichia coli expressing mcr-1 from dairy cows in China.

Objectives: To investigate the prevalence and molecular characteristics of ESBL-producing Escherichia coli (ESBL-EC) in faecal samples from dairy cows in China. Methods: In total, 651 faecal samples were collected from cows distributed among the 10 provinces of China. Potential ESBL-EC isolates were cultured on selective medium. The clonal relatedness of the ESBL-EC isolates was assessed using MLST. WGS was conducted on 3 mcr-positive isolates and 14 additional randomly selected ESBL-EC isolates. Southern blot, S1-PFGE and conjugation were performed for mcr-1-carrying isolates. The genetic environment of the pMCR-JLF4 plasmid was also analysed. Results: In total, 290 unique ESBL-EC isolates were detected from 284 cows (43.6%). Alleles of CTX-M were observed in 94.1% (273/290) of all isolates. The most prevalent genotypes observed in this study were blaCTX-M-14, blaCTX-M-15, blaCTX-M-17 and blaCTX-M-55. Differentiation of 79 STs with a polyclonal structure was accomplished using MLST. Clonal complex 10 was the most prevalent major complex detected here. Furthermore, the mcr-1 gene was detected in three isolates. The complete sequence of the mcr-1-containing pMCR-JLF4 was determined. The plasmid was 66.7 kb in length, with a genetic structure of nikA-nikB-mcr-1-pap2. Conjugation analysis confirmed that the mcr-1 gene in pMCR-JLF4 was transferable without the assistance of the ISApl1 gene. Conclusions: The data presented here suggest high prevalence of ESBL-EC in Chinese cow farms. Furthermore, it was clearly demonstrated that commensal E. coli strains can be reservoirs of blaCTX-M genes, potentially contributing to the dissemination and transfer of the mcr-1 gene to pathogenic bacteria among cows.

Comparative transcriptomic analysis reveals adriamycin-induced apoptosis via p53 signaling pathway in retinal pigment epithelial cells.

OBJECTIVE: This paper applied a transcriptomic approach to investigate the mechanisms of adriamycin (ADR) in treating proliferative vitreoretinopathy (PVR) using ARPE-19 cells. METHODS: The growth inhibitory effects of ADR on ARPE-19 cells were assessed by sulforhodamine B (SRB) assay and propidium iodide (PI) staining using flow cytometry. The differentially expressed genes between ADR-treated ARPE-19 cells and normal ARPE-19 cells and the signaling pathways involved were investigated by microarray analysis. Mitochondrial function was detected by JC-1 staining using flow cytometry and the Bcl-2/Bax protein family. The phosphorylated histone H2AX (γ-H2AX), phosphorylated checkpoint kinase 1 (p-CHK1), and phosphorylated checkpoint kinase 2 (p-CHK2) were assessed to detect DNA damage and repair. RESULTS: ADR could significantly inhibit ARPE-19 cell proliferation and induce caspase-dependent apoptosis in vitro. In total, 4479 differentially expressed genes were found, and gene ontology items and the p53 signaling pathway were enriched. A protein-protein interaction analysis indicated that the TP53 protein molecules regulated by ADR were related to DNA damage and oxidative stress. ADR reduced mitochondrial membrane potential and the Bcl-2/Bax ratio. p53-knockdown restored the activation of c-caspase-3 activity induced by ADR by regulating Bax expression, and it inhibited ADR-induced ARPE-19 cell apoptosis. Finally, the levels of the γ-H2AX, p-CHK1, and p-CHK2 proteins were up-regulated after ADR exposure. CONCLUSIONS: The mechanism of ARPE-19 cell death induced by ADR may be caspase-dependent apoptosis, and it may be regulated by the p53-dependent mitochondrial dysfunction, activating the p53 signaling pathway through DNA damage.