One-year results for myopia control of orthokeratology with different back optic zone diameters: a randomized trial using a novel multispectral-based topographer.

AIM: To present the 1-year results of a prospective cohort study investigating the efficacy, potential mechanism, and safety of orthokeratology (ortho-k) with different back optic zone diameters (BOZD) for myopia control in children. METHODS: This randomized clinical study was performed between Dec. 2020 and Dec. 2021. Participants were randomly assigned to three groups wearing ortho-k: 5 mm BOZD (5-MM group), 5.5 mm BOZD (5.5-MM group), and 6 mm BOZD (6-MM group). The 1-year data were recorded, including axial length, relative peripheral refraction (RPR, measured by multispectral refractive topography, MRT), and visual quality. The contrast sensitivity (CS) was evaluated by CSV-1000 instrument with spatial frequencies of 3, 6, 12, and 18 cycles/degree (c/d); the corneal higher-order aberrations (HOAs) were measured by iTrace aberration analyzer. The one-way ANOVA was performed to assess the differences between the three groups. The correlation between the change in AL and RPR was calculated by Pearson's correlation coefficient. RESULTS: The 1-year results of 20, 21, and 21 subjects in the 5-MM, 5.5-MM, and 6-MM groups, respectively, were presented. There were no statistical differences in baseline age, sex, or ocular parameters between the three groups (all P>0.05). At the 1-year visit, the 5-MM group had lower axial elongation than the 6-MM group (0.07±0.09 vs 0.18±0.11 mm, P=0.001). The 5-MM group had more myopic total RPR (TRPR, P=0.014), with RPR in the 15°-30° (RPR 15-30, P=0.015), 30°-45° (RPR 30-45, P=0.011), temporal (RPR-T, P=0.008), and nasal area (RPR-N, P<0.001) than the 6-MM group. RPR 15-30 in the 5.5-MM group was more myopic than that in the 6-MM group (P=0.002), and RPR-N in the 5-MM group was more myopic than that in the 5.5-MM group (P<0.001). There were positive correlations between the axial elongation and the change in TRPR (r=0.756, P<0.001), RPR 15-30 (r=0.364, P=0.004), RPR 30-45 (r=0.306, P=0.016), and RPR-N (r=0.253, P=0.047). The CS decreased at 3 c/d (P<0.001), and the corneal HOAs increased in the 5-MM group (P=0.030). CONCLUSION: Ortho-k with 5 mm BOZD can control myopia progression more effectively. The mechanism may be associated with greater myopic shifts in RPR.

IP3R2-mediated Ca2+ release promotes LPS-induced cardiomyocyte pyroptosis via the activation of NLRP3/Caspase-1/GSDMD pathway.

Pyroptosis plays a crucial role in sepsis, and the abnormal handling of myocyte calcium (Ca2+) has been associated with cardiomyocyte pyroptosis. Specifically, the inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) is a Ca2+ release channel in the endoplasmic reticulum (ER). However, the specific role of IP3R2 in sepsis-induced cardiomyopathy (SIC) has not yet been determined. Thus, this study aimed to investigate the underlying mechanism by which IP3R2 channel-mediated Ca2+ signaling contributes to lipopolysaccharide (LPS)-induced cardiac pyroptosis. The SIC model was established in rats by intraperitoneal injection of LPS (10 mg/kg). Cardiac dysfunction was assessed using echocardiography, and the protein expression of relevant signaling pathways was analyzed using ELISA, RT-qPCR, and western blot. Small interfering RNAs (siRNA) and an inhibitor were used to explore the role of IP3R2 in neonatal rat cardiomyocytes (NRCMs) stimulated by LPS in vitro. LPS-induced NLRP3 overexpression and GSDMD-mediated pyroptosis in the rats' heart. Treatment with the NLRP3 inhibitor MCC950 alleviated LPS-induced cardiomyocyte pyroptosis. Furthermore, LPS increased ATP-induced intracellular Ca2+ release and IP3R2 expression in NRCMs. Inhibiting IP3R activity with xestospongin C (XeC) or knocking down IP3R2 reversed LPS-induced intracellular Ca2+ release. Additionally, inhibiting IP3R2 reversed LPS-induced pyroptosis by suppressing the NLRP3/Caspase-1/GSDMD pathway. We also found that ER stress and IP3R2-mediated Ca2+ release mutually regulated each other, contributing to cardiomyocyte pyroptosis. IP3R2 promotes NLRP3-mediated pyroptosis by regulating ER Ca2+ release, and the mutual regulation of IP3R2 and ER stress further promotes LPS-induced pyroptosis in cardiomyocytes.

Orthokeratology with increased compression factor in adolescent myopia control: a 2-year prospective randomized clinical trial.

AIM: To explore the long-term efficacy, safety, and optical mechanism of orthokeratology with increased compression factor in adolescent myopia control. METHODS: A prospective, double-masked, and randomized clinical trial was performed from May 2016 to June 2020. Subjects aged between 8 and 16y, with myopia (-5.00 to -1.00 D), low astigmatism (≥-1.50 D) and anisometropia (≤1.00 D), were stratified into low (-2.75 to -1.00 D) and moderate (-5.00 to -3.00 D) myopia groups. Then they were randomly assigned to wear either increased compression factor (ICF; 1.75 D) orthokeratology or conventional compression factor (CCF; 0.75 D) orthokeratology. The data were recorded including axial length (AL), spherical equivalent (SE), best corrected visual acuity (BCVA), near visual acuity (NVA), corneal staining (using Efron grading scales), corneal hysteresis (CH), corneal resistance factor (CRF), higher-order aberrations (HOAs, expressed as root mean square, RMSh), and subfoveal choroidal thickness (SFChT) in the 2-year follow-up period. Pearson's correlation coefficient was conducted to analyze the association between the changes in AL and RMSh, SFChT. RESULTS: At the 2-year visit, there were no statistical differences in all the parameters between the ICF group and the CCF group in low myopia subjects (P>0.05). For the moderate myopia subjects, the ICF group had shorter AL elongation (0.23±0.08 vs 0.30±0.11 mm, P=0.015), higher RMSh (1.94±0.50 vs 1.65±0.51 µm, P=0.041), and higher SFChT (279.04±35.72 vs 254.08±29.60 µm, P=0.008) than those in CCF group. The change in AL was negatively correlated with RMSh (r=-0.687, P<0.001) and SFChT (r=-0.464, P=0.013). CONCLUSION: ICF orthokeratology can control the progression of moderate myopia more effectively, which might be related to greater RMSh and SFChT.

A miniaturized spatial temperature gradient capillary electrophoresis system with radiative heating and automated sample introduction for DNA mutation detection.

A miniaturized spatial temperature gradient CE system with automated sample introduction for DNA mutation detection was established. Continuous electrokinetic sample injection was achieved by combining an automated slotted-vial array sample introduction device to the spatial temperature gradient CE system. The temperature gradient was produced by a radiative heating system with a single graphite block heater, and the stability of the temperature gradient was investigated. The temperature variation of each measure point was 0.12-0.21% RSD (n=7) within 6 h. A 14-cm Teflon AF-coated silica capillary was used both as the separation channel and as the liquid-core waveguide tube of fluorescence signal. Under a temperature gradient from 54.8 to 59.5°C, a low range control mutation standard (209 bp) was separated within 4 min with only 5.6 nL sample consumption. Automated continuous sample introducing and changing were realized with a carryover of 3.3%. Utility of the system was further demonstrated by detecting K-ras gene mutations in paraffin tissue sections from two colorectal cancer patients.

An automated capillary electrophoresis system for high-speed separation of DNA fragments based on a short capillary.

A high-speed DNA fragment separation system was developed based on a short capillary and a slotted-vial array automated sample introduction system. The injection process of DNA sample in a short capillary was investigated systematically with three injection techniques including constant-field-strength, low-field-strength and translational spontaneous injections. Under the optimized conditions, picoliter-scale sample plugs (corresponding to ca. 20-µm plug length) were obtained, which ensure the high-speed and high-efficiency separation for DNA fragments with a short effective separation length. Other separation conditions including the sieving matrix concentration, separation field strength and effective separation length were also optimized. The present system was applied in the separation of ΦX174-Hae III digest DNA marker. With an effective separation length of 2.5 cm, the separation could be achieved in <100 s with plate heights ranging from 0.21 to 0.74 µm (corresponding to plate numbers from 4.86 × 10(6) to 1.36 × 10(6)/m). The repeatabilities for the migration time of the eleven fragments were between 0.4 and 1.1% RSD (n=8). By using the automated continuous injection method, the separation for four different DNA samples could be achieved within 250 s. The present system was further applied in the fast sizing of real DNA samples of PCR products.

DNA mutation detection with chip-based temperature gradient capillary electrophoresis using a slantwise radiative heating system.

A simple and robust chip-based temperature gradient capillary electrophoresis (TGCE) system was developed for DNA mutation/single-nucleotide polymorphism (SNP) analysis using a radiative heating system. Reproducible, stable and uniform temperature gradients were established along a 3 cm length of the electrophoretic separation channel using a single thermostated aluminium heater plate. The heater was slightly slanted relative to the plane of the glass chip at 0.2-1.3 degrees by inserting thin spacers between the plate and chip at one end to produce differences in radiative heating that created the temperature gradient. On-chip TGCE analyses of 4 mutant DNA model samples amplified from plasmid templates, each containing a single base substitution, with a wide range of melting temperatures, showed that mutations were successfully detected under a wide temperature gradient of 10 degrees C and within a short gradient region of about 3 cm (3.3 degrees C cm(-1) gradient). The radiative heating system was able to establish stable spatial temperature gradients along short microfluidic separation channels using simple peripheral equipment and manipulation while ensuring good resolution for detecting a wide range of mutations. Effectiveness of the system was demonstrated by the successful detection of K-ras gene mutations in 6 colon cancer cell lines.

DNA purification on a lab-on-valve system incorporating a renewable microcolumn with in situ monitoring by laser-induced fluorescence.

Bead injection in a lab-on-valve (LOV) system was adopted for DNA purification via micro solid-phase extraction (SPE) with a renewable silica microcolumn packed in a channel of the LOV unit. The complex matrix components in human whole blood, including proteins, were well eliminated by choosing properly the sample loading and elution media. The DNA purification process was monitored on-line by using laser-induced fluorescence in a demountable side part of the LOV unit incorporating optical fibers. The practical applicability of the entire system was demonstrated by separation/purification of lambda-DNA in a simulated matrix and human blood genetic DNA by performing SPE, in situ monitoring of the purified products, and postcolumn PCR amplification. When DNAs in a simulated matrix (10.0 ng microl-1 lambda-DNA, 50 ng microl-1 bovine serum albumin, 1.0% Triton X-100) were processed in the present system and laser-induced fluorescence was monitored at 610 nm, an overall extraction/collection efficiency of 70% was achieved by employing identical sample loading and an elution flow rate of 0.5 microl s-1, along with a precision of 3.8% relative standard deviation. DNA separation and purification from human whole-blood samples were performed under similar conditions.

[Study on the relationship between copper, lysyl oxidase and premature rupture of membranes].

OBJECTIVE: To determine the relationships between maternal serum copper, amniotic copper, lysyl oxidase (LOX) and collagen III in pregnant women with premature rupture of membranes (PROM) and without PROM. METHODS: One hundred women with PROM were enrolled in this study, and divided into 37-42 weeks, 34-36(+6) weeks and 28-33(+6) weeks according to gestational age. One hundred non-PROM pregnancies matching the same gestational ages were recruited as control group. Copper of maternal serum and amnion in two groups were compared by FAAS method. Amniotic LOX was analyzed by fluorometry. Amniotic collagen III was detected by immunohistochemical method and computer image analysis system (absorbance, A). Linear correlation analysis was used to explore the relationships between maternal serum copper, amniotic copper, LOX and collagen III. RESULTS: (1) For 37-42 weeks pregnant women, serum copper was correlated positively with amniotic copper in two groups, r = 0.82 (P < 0.001), but for other parameters, the serum and amniotic levels had no correlations. (2) For 34-36(+6) and 28 - 33(+6) weeks pregnant women, there were positive correlations between serum copper, amniotic copper, LOX and collagen III in two groups (P < 0.01). (3) For 37-42 weeks pregnant women, serum copper and amniotic copper had no difference between two groups (P > 0.05), but amniotic LOX and collagen III decreased significantly compared with controls, being [(0.53 +/- 0.10) microg/g vs (0.75 +/- 0.10) microg/g, P < 0.01], (0.36 +/- 0.01 vs 0.37 +/- 0.01, P < 0.05) respectively. (4) For 34 - 36(+6) weeks pregnant women, serum copper, amniotic copper, LOX and collagen III reduced obviously between two groups (P < 0.01), being respectively [(115.23 +/- 9.56) mg/L vs (139.03 +/- 10.59) mg/L], [(0.21 +/- 0.04) microg/mg vs (0.29 +/- 0.04) microg/mg], [(0.54 +/- 0.10) microg/g vs (0.70 +/- 0.13) microg/g], and (0.36 +/- 0.01 vs 0.37 +/- 0.01). (5) For 28-33(+6) weeks pregnant women, serum copper, amniotic copper, LOX and collagen III also reduced obviously between two groups (P < 0.01), [(120.31 +/- 8.04) microg/L vs (136.40 +/- 8.21) microg/L], [(0.21 +/- 0.04) microg/mg vs (0.26 +/- 0.03) microg/mg], [(0.62 +/- 0.09) microg/g vs (0.72 +/- 0.09) microg/g], and (0.35 +/- 0.01 vs 0.38 +/- 0.01). CONCLUSION: For term pregnant women, the decreases of amniotic LOX and collagen III in PROM group are not affected by serum copper, perhaps by excessive degeneration of amnion; for non-term pregnant women, the decreases of serum copper, amniotic copper, LOX and collagen III are correlated directly with PROM.