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06). After that, T&E group maintained this tendency, reaching the lowest CFT value at 36 months, whereas PRN group showed an increased in CFT values (PRN -104 [-807.7 to +297] μm and T&E -103 [-575 to +244], μm p=0.63). Patients treated with T&E regimen received a significantly higher number of injections (PRN 16.3±7.6 vs T&E 23.9 ±9.4, p less then 0.01). CONCLUSION Our results demonstrated a trend towards for T&E to achieve higher marks in BCVA, peaking at 12 months, and lower CFT thickness at the end of three years. Despite the higher number of injections performed in the T&E group the mean BCVA reverts to baseline values at 3 years. BACKGROUND Whether a transumbilical or periumbilical incision is beneficial for the initial peritoneal access in laparoscopic abdominal surgery has been debated. Our aim is to determine whether a transumbilical or periumbilical incision is a better route for the initial umbilical trocar. METHODS PubMed, Embase, and Cochrane Library databases were searched for articles published before March 2020. The meta-analysis calculated the pooled effect size by using a random effects model. RESULTS Five trials involving 783 patients were reviewed. The transumbilical group significantly reduced operation time (mean difference -7.73; 95% confidence interval -13.10 to -2.35) when compared to the periumbilical group. The length of hospital stay, mean pain scores on operation day and postoperation day 1 did not differ significantly between the two groups. Moreover, the incidence of surgical site infection, cosmetic satisfaction, and complication rate did not differ significantly between groups. CONCLUSION A transumbilical incision is better than a periumbilical incision for laparoscopic surgery as it saves operation time. Hence, we suggest transumbilical incisions for the initial peritoneal access in laparoscopic abdominal surgery. Solvent-free analysis of organic pollutants from a complex matrix has attracted significant attention. Selleck AG 825 In this work, we designed and fabricated a facile cooling-assisted solid-phase microextraction device (CA-SPME) that could be used for the solvent-free extraction of volatile and semivolatile compounds from a complex matrix using a commercial SPME probe. Determination of polycyclic aromatic hydrocarbons (PAHs) from soil samples was carried out to evaluate the performance of the designed CA-SPME device. The effects of heating temperature, cooling temperature, extraction time, and moisture content in soil on extraction efficiency were investigated. To enhance the extraction efficiency of the targeted analytes, the matrix solid-phase dispersion technique (MSPD) by grinding soil with silica gels was applied. The results showed that grinding with silica gels promoted the release of PAHs from the soil matrix to the headspace by disrupting the matrix structure, which then facilitated the extraction efficiency. Under optimized conditions, for all of the targeted PAHs, the method exhibited good linearity (40-4000 ng g-1) with regression coefficients (R2) ranging from 0.9586 to 0.9964. The limits of detection and limits of quantification ranged from 4.2 to 8.5 ng g-1 and 14.0-28.5 ng g-1, respectively. Relative standard deviations corresponding to the analysis of spiked soil samples (n = 5) were 8.1-13.4%. The solvent-free analysis of the certified soil sample using the proposed method demonstrated satisfactory results. Nucleic acids are important biomarkers for disease detection, monitoring, and treatment. Advances in technologies for nucleic acid analysis have enabled discovery and clinical implementation of nucleic acid biomarkers. However, challenges remain with technologies for nucleic acid analysis, thereby limiting the use of nucleic acid biomarkers in certain contexts. Here, we review single-molecule technologies for nucleic acid analysis that can be used to overcome these challenges. We first discuss the various types of nucleic acid biomarkers important for clinical applications and conventional technologies for nucleic acid analysis. We then discuss technologies for single-molecule in vitro and in situ analysis of nucleic acid biomarkers. Finally, we discuss other ultra-sensitive techniques for nucleic acid biomarker detection. Monitoring cellular GSH dynamics is of great value to understand its complex biological functions and related diseases. It still remains challenging to understand the mechanism of GSH dynamics in complex intracellular environment. Herein, a novel fluorescent probe featured with chlorinated coumarin-TCF was exploited for sensing of GSH with high selectivity and high sensitivity. Large fluorescence enhancement at 471 nm was observed upon addition of GSH with large emission distance (Δλem = 219 nm). This novel probe was successfully applied to monitor endogenous and exogenous GSH dynamics without interference of other thiols via fluorescence imaging. More importantly, using this probe, low dose reactive oxygen species induced GSH increasement through nuclear factor (erythroid-derived 2)-like 2 (Nrf2) signal pathway in BEL-7402 cells was observed. This novel fluorescent probe has the potential for quantitative monitoring of cellular GSH, which will promote further understanding of its physiological and pathological roles in biological systems. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) has become a popular method for analysis of the conformational dynamics and interactions of proteins. Disulfide-bonded proteins, however, present a challenge to HDX-MS as they require efficient disulfide bond reduction prior to enzymatic proteolysis. Electrochemical reduction (ER) provides an attractive solution to tackle disulfide-bonded proteins that are resistant to conventional chemical reduction during HDX-MS. However, ER-enabled HDX-MS has been limited by technical challenges including partial unwanted protein oxidation side-reactions, incompatibility with certain buffer components and most importantly, a lack of overall method robustness. In this study, we have sought to address these challenges. We perform a systematic screening of the compatibility of ER to buffers commonly used in HDX-MS samples by using a reliable and simple system suitability test (SST). Furthermore, we demonstrate the benefits of a new design of the electrochemical cell (EC) for ER-enabled HDX-MS, which include a) high repeatability and robustness over large sample batches without the need for electrode polishing and b) high reduction efficiency of disulfide-bonded proteins without unwanted oxidation side-reactions.