Reducing Aerosol Emissions by Decoupled Parameter Management during the CO2 Capture Process in a Multi-stage Circulation Absorber.

Aerosol emissions from the CO2 capture process have a significant impact in terms of solvent loss and environmental pollution. Here, we propose a novel approach with multi-stage circulation for CO2 capture and synergistic aerosol reduction, which divides the absorption section into three circulation stages and reduces aerosol emissions through decoupled operation of the three absorption sections and the management of solvent CO2 loadings. Experimental results show that with the decoupled management of the liquid-gas ratio and solvent temperature in absorption sections, the aerosol mass concentration at the outlet of the 3rd absorption section can be reduced by 25.6% to a minimum of 349.7 mg/m3 at a liquid-gas ratio of 43.2 L/m3 and a solvent temperature of 303 K. Furthermore, aerosol removal is performed by setting up a water wash section after the absorption section. The aerosol mass concentration at the outlet of the absorber is reduced to 168.6 mg/m3 with the regulation of the wash water temperature and flow rate. In addition, improvements are proposed for the combination of the utilization of recovered solvents and the co-removal of SO2. This study provides innovative insights into the design of the CO2 capture system and the reduction of aerosol emissions, which are of great significance for the mitigation of global warming and the control of environmental pollution.

Meibomian gland alteration in patients with systemic lupus erythematosus.

PURPOSE: To investigate meibomian gland (MG) alteration in patients with systemic lupus erythematosus (SLE). METHODS: This study included 23 SLE patients evaluated with Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) and 21 healthy controls (HCs). All the subjects were evaluated with Ocular Surface Disease Index (OSDI) questionnaire, and the eyes were performed examinations of tear meniscus height (TMH), non-invasive keratographic tear film break-up time (NIKBUT), Schirmer I Test, MG eyelid score, meibography score, and in vivo confocal microscopy (IVCM) on the meibomian gland. RESULTS: There was no significant difference between the SLE patients and the HCs in the TMH, NIKBUT, and Schirmer I Test. However, the SLE patients had higher MG eyelid scores and meibography scores on both upper eyelid and lower eyelid than the HCs. Through meibography observation, 34.8% of the SLE patients presented MG deficiency in Grade 3, whereas that of all the HCs were less than Grade 3. The SLE patients were found to have significant MG atrophy and vascular enrichment around the meibomian glands (MGs). The SLE patients were also found to have excessive inflammatory cell infiltration around the MGs, especially the typical lymph node-like foci of inflammatory cell infiltration. CONCLUSIONS: MG alteration can be found in the SLE patients. Examinations of the MGs can help diagnose or infer ocular diseases at an early stage of SLE.

Effects of ticagrelor monotherapy vs. clopidogrel monotherapy on platelet reactivity: a randomized, crossover clinical study (SINGLE study).

Increasing clinical trials demonstrated that the discontinuation of aspirin while maintaining a P2Y12 inhibitor monotherapy could decrease the risk of bleeding without losing the antithrombotic effect. However, no data are available on the platelet reactivity of patients undergoing ticagrelor monotherapy vs. clopidogrel. Therefore, we performed this study to observe the efficacy of ticagrelor monotherapy vs. clopidogrel in Chinese patients with chronic coronary syndrome. This randomized, single-blinded, crossover trial enrolled 50 patients who were administered with ticagrelor (90 mg twice daily for 2 weeks) or clopidogrel (75 mg once daily for 2 weeks). Followed by a 2-week washout period, the two groups of patients underwent a crossover trial. Light transmission aggregometry (LTA) and thromboelastography (TEG) assays were used to test platelet reactivity. The platelet aggregation rate (PAgR) of ADP and AA was significantly lower with ticagrelor than clopidogrel (PAgR of ADP, 27.30% (7.30%-42.635%) vs. 35.55% (12.03%-69.25%), P = .0254; PAgR of AA, 77.80% (21.60%-86.43%) vs. 83.10% (67.13%-87.20%), P = .0400). There was no significant difference in PAgR of collagen and epinephrine between the two groups. The TEG assay showed that ADP and AA, which induced the inhibition of platelet aggregation, were significantly higher in the ticagrelor group than those in the clopidogrel group [ADP%, 69.00% (59.68%-88.95%) vs. 60.55% (35.98%-78.35%), P = .0020; AA%, 53.65% (22.75%-79.28%) vs. 15.15% (5.75%-70.25%), P = .0127]. High on-treatment platelet reactivity (HTPR) on ADP was 2.17% with ticagrelor and 19.57% with clopidogrel. HTPR on AA was 50.00% with ticagrelor and 69.57% with clopidogrel. Ticagrelor and clopidogrel caused the inhibition of ADP and AA-induced platelet aggregation. Moreover, ticagrelor monotherapy had a stronger inhibitory effect than clopidogrel monotherapy (except on collagen and epinephrine).

High-Fat Diet-Induced Functional and Pathologic Changes in Lacrimal Gland.

The lacrimal gland is critical for maintaining the homeostasis of the ocular surface microenvironment through secreting aqueous tears in mammals. Many systemic diseases such as Sjögren syndrome, rheumatoid arthritis, and diabetes can alter the lacrimal gland function, eventually resulting in aqueous tear-deficient dry eye. Here, a high-fat diet (HFD) experimental mouse model was used to clarify how hyperlipidemia affects lacrimal gland function. Aqueous tear secretion fell about 50% after 1 month on a HFD. Lipid droplets accumulated in the matrix and acinar cells of the lacrimal gland after this period, along with changes in the lipid metabolism, changes in gene expression levels, and disruption of fatty acid oxidative activity. Immune cell infiltration and rises in the gene expression levels of the inflammation-related cytokines Il1ß, Tnfα, Tsg6, Il10, Mmp2, and Mmp9 were found. HFD also induced mitochondrial hypermegasoma, increased apoptosis, and decreased lacrimal gland acinar cell proliferation. Replacement of the HFD with the standard diet partially reversed pathologic changes in the lacrimal gland. Similarly, supplementing the HFD with fenofibrate also partially reversed the inhibited tear secretion and reduced lipid accumulation, inflammation, and oxidative stress levels. The authors conclude that a HFD induces pathophysiological changes and functional decompensation of the lacrimal gland. Therefore, ingestion of a HFD may be a causative factor of dry eye disease.

A miRNA-Encoded Small Peptide, vvi-miPEP171d1, Regulates Adventitious Root Formation.

One of the biggest challenges in clonal propagation of grapevine (Vitis vinifera) is difficulty of rooting. Adventitious root initiation and development are the critical steps in the cutting and layering process of grapevine, but the molecular mechanism of these processes remains unclear. Previous reports have found that microRNA (miRNA)-encoded peptides (miPEPs) can regulate plant root development by increasing the transcription of their corresponding primary miRNA. Here, we report the role of a miPEP in increasing adventitious root formation in grapevine. In this study, we performed a global analysis of miPEPs in grapevine and characterized the function of vvi-miPEP171d1, a functional, small peptide encoded by primary-miR171d. There were three small open reading frames in the 500-bp upstream sequence of pre-miR171d. One of them encoded a small peptide, vvi-miPEP171d1, which could increase the transcription of vvi-MIR171d Exogenous application of vvi-miPEP171d1 to grape tissue culture plantlets promoted adventitious root development by activating the expression of vvi-MIR171d Interestingly, neither exogenous application of the vvi-miPEP171d1 peptide nor overexpression of the vvi-miPEP171d1 coding sequence resulted in phenotypic changes in Arabidopsis (Arabidopsis thaliana). Similarly, application of synthetic ath-miPEP171c, the small peptide encoded by the Arabidopsis ortholog of vvi-MIR171d, inhibited the growth of primary roots and induced the early initiation of lateral and adventitious roots in Arabidopsis, while it had no effect on grape root development. Our findings reveal that miPEP171d1 regulates root development by promoting vvi-MIR171d expression in a species-specific manner, further enriching the theoretical research into miPEPs.

Chemical Information Exchange in Organized Protocells and Natural Cell Assemblies with Controllable Spatial Positions.

An ultrasound-based platform is established to prepare homogenous arrays of giant unilamellar vesicles (GUVs) or red blood cell (RBCs), or hybrid assemblies of GUV/RBCs. Due to different responses to the modulation of the acoustic standing wave pressure field between the GUVs and RBCs, various types of protocell/natural cell hybrid assemblies are prepared with the ability to undergo reversible dynamic reconfigurations from vertical to horizontal alignments, or from 1D to 2D arrangements. A two-step enzymatic cascade reaction between transmitter glucose oxidase-containing GUVs and peroxidase-active receiver RBCs is used to implement chemical signal transduction in the different hybrid micro-arrays. Taken together, the obtained results suggest that the ultrasound-based micro-array technology can be used as an alternative platform to explore chemical communication pathways between protocells and natural cells, providing new opportunities for bottom-up synthetic biology.

Evolution of Condensable Fine Particle Size Distribution in Simulated Flue Gas by External Regulation for Growth Enhancement.

Condensation fine particles (CFPs) from coal-fired flue gas harm humans and the environment after being emitted into the atmosphere. Given their small size (<0.1 µm), difficulty arises in efficiently removing CFPs by wet electrostatic precipitators and mist eliminators. In this work, a laboratory apparatus was used to study the CFP growth under simulated power plant conditions. Four methods were independently investigated to increase the particle size: addition of ammonia, addition of fly ash, decreasing temperature, and applying an electrical discharge. Results demonstrated that the CFP size distribution possessed a unimodal structure with peak at 0.05 µm. At increased ammonia concentration from 10 to 30 ppm, the peak of growth factor shifted rightward and increased from 1.21 to 1.35 and the range of growth factor >1 was significantly broadened due to joint action of multiple mechanisms. Fly ash acted as the core, and CFPs adhered to the ash surface when forming ash-salt droplets. Cooling flue gas could also enhance the CFP growth due to vapor condensation. At decreased temperature from 45 to 30 °C, the median diameter of CFPs increased by 15%. Finally, the growth and agglomeration of CFPs can be further enhanced when an external electrical field was utilized. The size range of growth factor >1 can be broadened, and the peak growth factor significantly increased at 8 kV applied voltage. The research findings provide valuable guidance for effectively improving the CFP removal efficiency by external regulation for growth enhancement.

Comparative Analysis of miRNA Abundance Revealed the Function of Vvi-miR828 in Fruit Coloring in Root Restriction Cultivation Grapevine (Vitis vinifera L.).

Root restriction cultivation leads to early maturation and quality improvement, especially in the anthocyanin content in grapevine. However, the molecular mechanisms that underlie these changes have not been thoroughly elucidated. In this study, four small RNA libraries were constructed, which included the green soft stage (GS) and ripe stage (RS) of 'Muscat' (Vitis vinifera L.) grape berries that were grown under root restriction (RR) and in traditional cultivation (no root restriction, CK). A total of 162 known miRNAs and 14 putative novel miRNAs were detected from the four small RNA libraries by high-throughput sequencing. An analysis of differentially expressed miRNAs (DEMs) revealed that 13 miRNAs exhibited significant differences in expression between RR and CK at the GS and RS stages, respectively. For different developmental stages of fruit, 23 and 34 miRNAs showed expression differences between the GS and RS stages in RR and CK, respectively. The expression patterns of the eight DEMs and their targets were verified by qRT-PCR, and the expression profiles of target genes were confirmed to be complementary to the corresponding miRNAs in RR and CK. The function of Vvi-miR828, which showed the down regulated expression in the RS stage under root restriction, was identified by gene transformation in Arabidopsis. The anthocyanin content significantly decreased in transgenic lines, which indicates the regulatory capacity of Vvi-miR828 in fruit coloration. The miRNA expression pattern comparison between RR and CK might provide a means of unraveling the miRNA-mediated molecular process regulating grape berry development under root restricted cultivation.

Fractional order neural sliding mode control based on the FO-Hammerstein model of piezoelectric actuator.

Piezoelectric actuators (PEA) are extensively utilized in high-precision micro-measurement and operation. However, owing to the rate-dependent nature of its hysteresis, its accuracy in certain applications would suffer significantly, and the system would become unstable. To address this issue, a new method for developing a feedback control system that can reduce the rate-dependent impacts of PEA on the positioning system is provided using PEA as the research object. This strategy is based on the fractional order Hammerstein model (FO-Hammerstein). For the fractional order model, a novel fractional order integral sliding mode surface is proposed here that can accurately delineate the dynamic characteristics of PEA. This sliding mode surface is composed of a fractional polynomial and an integral term, which can better minimize static errors and monitor reference signals, and it is built using a fractional neural sliding mode control (BP-FSMC) method. The control technique can be extensively utilized in various systems, such as FO-Hammerstein and those described by the fractional transfer function. The research introduces a neural network and an artificial bee colony algorithm (DeC-ABC) that are used to alter the system's parameters. The study's findings reveal that a system with high resilience can follow the signals from both composite and single input sources. Compared with the fractional order sliding mode control approach on the basis of extended state observer, the fractional order sliding mode control method based on single parameter adaptive law and the proportional integral sliding mode control method on the basis of feedforward compensation, this method has a quicker response time and lower tracking error.

Prediction of inlet SO2 concentration of wet flue gas desulfurization (WFGD) by operation parameters of coal-fired boiler.

Circulating fluidized bed (CFB) boilers with wet flue gas desulfurization (WFGD) system is a popular technology for SO2 removal in the coal-fired thermal power plant. However, the long response time of continues emission monitoring system (CEMS) and the hardness of continuously monitoring the coal properties leads to the difficulties for controlling WFGD. It is important to build a model that is adaptable to the fluctuation of load and coal properties, which can obtain the SO2 concentration ahead CEMS, without relying on coal properties. In this paper, a prediction model of inlet SO2 concentration of WFGD considering the delay between the features and target based on long-short term memory (LSTM) network with auto regression feature is established. The SO2 concentration can be obtained 90 s earlier than CEMS. The model shows good adaptability to the fluctuation of SO2 concentration and coal properties. The root-mean-squared error (RMSE) and R squared (R2) of the model are 30.11 mg/m3 and 0.986, respectively. Meanwhile, a real-time prediction system is built on the 220 t/h unit. A field test for long-term operation has been conducted. The prediction system is able to continuously and accurately predict the inlet SO2 concentration of the WFGD, which can provide the operators with an accurate reference for the control of WFGD.

Simulation investigation on marine exhaust gas SO2 absorption by seawater scrubbing.

Ships have become an important source of SO2 emission in coastal areas with the rapid development of maritime transport. It is of great significance to develop a marine scrubber for reducing SO2 emission of ships. In this study, numerical simulation of a full-scale marine spray scrubber is conducted to investigate two-phase flow pattern and SO2 absorption process in the scrubber. A desulfurization model based on seawater absorbent is coupled into the simulation, which considers the mass transfer between phases and seawater aqueous phase chemistry simultaneously. A distribution ring is introduced in the scrubber to enhance the desulfurization performance of the scrubber. The result of simulation shows that the distribution ring can optimize effectively the distribution of gas-liquid phases and enhance the SO2 absorption. Under vertical condition, the desulfurization efficiency could be promoted approximate 6% after installing a distribution ring. The inclined condition resulting from the ship swinging could lead to the uneven distribution of droplets and an obvious decrease (8.7%) of desulfurization efficiency, whereas the desulfurization performance of the scrubber could be ensured with a distribution ring installed even under an inclined condition. Finally, a spray scrubber design scheme has been developed and successfully applied in the exhaust gas cleaning system (EGCS) of a container ship. Test result shows the outlet average value of SO2/CO2 can be reduced to 3.55. Meanwhile, the consistency of test data and calculation result indicates the applicability of the numerical model established for the simulation and optimization of the scrubber in industrial applications also.Implications: EGCS is an effective method to reduce SO2 emission of marine industry. However, different from a land desulfurization tower, the application of a spray scrubber in EGCS faces more problems due to the different application scenarios and complex sea conditions (inclined condition resulting from ships swinging and so on) during sailing. In this work, a numerical model capable of investigating physical and chemical phenomena in the scrubber simultaneously is established, which can produce a great amount of data for the operation instruction of EGCS and the design and optimization of the marine spray scrubber. The distribution ring is introduced in the marine spray scrubber to intensify the SO2 absorption and enhance the desulfurization performance of the scrubber under different working conditions.

Dynamic NOx emission prediction based on composite models adapt to different operating conditions of coal-fired utility boilers.

An accurate NOx concentration prediction model plays an important role in low NOx emission control in power stations. Predicting NOx in advance is of great significance in satisfying stringent environmental policies. This study aims to accurately predict the NOx emission concentration at the outlet of boilers on different operating conditions to support the DeNOx procedure. Through mutual information analysis, suitable features are selected to build models. Long short-term memory (LSTM) models are utilized to predict NOx concentration at the boiler's outlet from selected input features and exhibit power in fitting multivariable coupling, nonlinear, and large time-delay systems. Moreover, a composite LSTM model composed of models on different operating conditions, like steady-state and transient-state condition, is prosed. Results of one whole day of typical operating data show that the accuracy of the NOx concentration and fluctuation trend prediction based on this composite model is superior to that using a single LSTM model and other non-time-sequence models. The root mean square error (RMSE) and R2 of the composite LSTM model are 3.53 mg/m3 and 0.89, respectively, which are better than those of a single LSTM (i.e., 5.50 mg/m3 and 0.78, respectively).

Ovalbumin-Induced Allergic Inflammation Diminishes Cross-Linked Collagen Structures in an Experimental Rabbit Model of Corneal Cross-Linking.

Background: Allergic conjunctivitis (AC) is one of the reported potential risk factors of progression in keratoconus patients after corneal cross-linking surgery; however, the causal relationship is still inconclusive. Recent studies have indicated that various inflammatory cytokines play a vital role in the development of primary keratoconus. It is still unclear whether these inflammatory mediators also trigger CXL failures. This study aimed to investigate the impact of AC on the rabbit corneas after trans-epithelial corneal cross-linking (TCXL). Methods: A total of six rabbits were kept untreated as the normal control (NC) group. A total of 18 rabbits were treated by TCXL and divided into three groups (six in each group), namely, no treatment (TCXL group); induction of AC (TCXL + AC group); and induction of AC plus topical prednisolone acetate (TCXL + AC + PA group), according to additional treatment. AC was induced by topical application of ovalbumin after intraperitoneal pre-sensitization with ovalbumin. Rabbits were evaluated by slit lamp, in vivo laser scanning confocal microscopy, anterior segment optical coherence tomography, and measurement of corneal biomechanics. The cornea specimens were collected for the transmission electron microscope, the collagenase I digestion test, and PCR assay for TNF-α, IL-6, IL-1ß, matrix metalloproteinase 9 (MMP-9), lysyl oxidase (LOX), and tissue inhibitor of metalloproteinases 1 (TIMP-1) on the day (D) 28. Results: On D28, the TNF-α, IL-6, IL-1ß, MMP-9, and LOX levels were significantly increased while the TIMP-1 was decreased in the TCXL + AC group when compared with the TCXL and TCXL + AC + PA groups. In vivo confocal microscopy revealed that at a depth of 150-210 µm, a trabecular patterned hyperdense structure surrounded by elongated needle-like processes could be observed in the TCXL and TCXL + AC + PA groups, but hardly seen in the TCXL + AC group. The demarcation lines were indistinct and blurred in the TCXL + AC group. An electron microscope demonstrated less interlacing fibril lamellae and higher interfibrillar spacing in the TCXL + AC group. The stability of corneal biomechanics and resistance to collagenase were decreased in the TCXL + AC group. Conclusion: The corneal microstructures induced by TCXL and biomechanical stability were diminished in rabbits with AC but could be maintained by topical anti-inflammatory treatment. Our results supported the causal relationship between altered cytokine profiles and corneal microstructure after primary corneal cross-linking.

Hyperglycemia Induces Meibomian Gland Dysfunction.

Purpose: Patients diagnosed with diabetes are inclined to have abnormalities on stability of tear film and disorder of meibomian gland (MG). This study aims to explore the pathological change of MG induced by diabetes in a rat model. Methods: Sprague-Dawley (SD) rats were intraperitoneally injected with streptozotocin (STZ) to establish a diabetic animal model. Lipid accumulation in MG was detected by Oil Red O staining and LipidTox staining. Cell proliferation status was determined by Ki67 and P63 immunostaining, whereas cell apoptosis was confirmed by TUNEL assay. Gene expression of inflammatory cytokines and adhesion molecules IL-1α, IL-1ß, ELAM1, ICAM1, and VCAM1 were detected by RT-PCR. Activation of ERK, NF-κB, and AMPK signaling pathways was determined by Western Blot analysis. Oxidative stress-related factors NOX4, 4HNE, Nrf2, HO-1, and SOD2 were detected by immunostaining or Western Blot analysis. Tom20 and Tim23 immunostaining and transmission electron microscopy were performed to evaluate the mitochondria functional and structure change. Results: Four months after STZ injection, there was acini dropout in MG of diabetic rats. Evident infiltration of inflammatory cells, increased expression of inflammatory factors, and adhesion molecules, as well as activated ERK and NF-κB signaling pathways were identified. Oxidative stress of MG was evident in 4-month diabetic rats. Phospho-AMPK was downregulated in MG of 2-month diabetic rats and more prominent in 4-month rats. After metformin treatment, phospho-AMPK was upregulated and the morphology of MG was well maintained. Moreover, inflammation and oxidative stress of MG were alleviated after metformin intervention. Conclusions: Long-term diabetes may lead to Meibomian gland dysfunction (MGD). AMPK may be a therapeutic target of MGD induced by diabetes.

Rate-Dependent Modeling of Piezoelectric Actuators for Nano Manipulation Based on Fractional Hammerstein Model.

Piezoelectric actuators (PEAs), as a smart material with excellent characteristics, are increasingly used in high-precision and high-speed nano-positioning systems. Different from the usual positioning control or fixed frequency tracking control, the more accurate rate-dependent PEA nonlinear model is needed in random signal dynamic tracking control systems such as active vibration control. In response to this problem, this paper proposes a Hammerstein model based on fractional order rate correlation. The improved Bouc-Wen model is used to describe the asymmetric hysteresis characteristics of PEA, and the fractional order model is used to describe the dynamic characteristics of PEA. The nonlinear rate-dependent hysteresis model can be used to accurately describe the dynamic characteristics of PEA. Compared with the integer order model or linear autoregressive model to describe the dynamic characteristics of the PEA Hammerstein model, the modeling accuracy is higher. Moreover, an artificial bee colony algorithm (DE-ABC) based on differential evolution was proposed to identify model parameters. By adding the mutation strategy and chaos search of the genetic algorithm into the previous ABC, the convergence speed of the algorithm is faster and the identification accuracy is higher, and the simultaneous identification of order and coefficient of the fractional model is realized. Finally, by comparing the simulation and experimental data of multiple sets of sinusoidal excitation with different frequencies, the effectiveness of the proposed modeling method and the accuracy and rapidity of the identification algorithm are verified. The results show that, in the wide frequency range of 1-100 Hz, the proposed method can obtain more accurate rate-correlation models than the Bouc-Wen model, the Hammerstein model based on integer order or the linear autoregressive model to describe dynamic characteristics. The maximum error (Max error) is 0.0915 µm, and the maximum mean square error (RMSE) is 0.0244.

A real-time optimization method for economic and effective operation of electrostatic precipitators.

Electrostatic precipitators (ESP) have been considered as the main particulate matter (PM) removal facility in the energy industry. This paper presents a real-time optimization method for a one-chamber industrial ESP in an ultra-low emission power plant with an intelligent optimization system (IOS). The IOS seeks to optimize the energy consumption of ESP subject to the outlet concentration requirement in real-time. A coordination control logic is designed to regulate the optimized operation set points with varying operation conditions. The operation optimized by the IOS is compared with the operations under PID (proportion-integral-derivative) and manual control. The results show that the IOS improves the emission compliance rate from 95% of manual control to 100% and the medium concentration is tuned to be 46.6% closer to the emission target. Furthermore, a good balance between emission and energy consumption is achieved, with 35.50% energy conservation for the same emission upper limit of 30 mg/m3. These results prove that the IOS significantly contributes to the efficient operation and economic PM removal by ESP for the energy industry. IMPLICATIONS: Electrostatic precipitators (ESP) is one of the main PM removal facilities in coal-fired power plants. An intelligent optimization system (IOS) with prediction, optimization, and control modules is designed and constructed for the ESP in an ultra-low emission power plant. A PM removal model is used to predict the outlet concentration of the ESP. The optimal energy consumption of ESP subject to the outlet concentration requirement problem is solved by the particle swarm optimization. A closed-loop and rapping tolerant method is used to eliminate the fluctuation in time-averaged concentration. The system raised is able to ensure the compliance rate while decreasing the energy consumption of the ESP.

Tissue engineered corneal epithelium derived from clinical-grade human embryonic stem cells.

PURPOSE: To construct tissue engineered corneal epithelium from a clinical-grade human embryonic stem cells (hESCs) and investigate the dynamic gene profile and phenotypic transition in the process of differentiation. METHODS: A stepwise protocol was applied to induce differentiation of clinical-grade hESCs Q-CTS-hESC-1 and construct tissue engineered corneal epithelium. Single cell RNA sequencing (scRNA-seq) analysis was performed to monitor gene expression and phenotypic changes at different differentiation stages. Immunostaining, real-time quantitative PCR and Western blot analysis were conducted to detect gene and protein expressions. After subcutaneous transplantation into nude mice to test the biosafety, the epithelial construct was transplanted in a rabbit corneal limbal stem cell deficiency (LSCD) model and followed up for eight weeks. RESULTS: The hESCs were successfully induced into epithelial cells. scRNA-seq analysis revealed upregulation of ocular surface epithelial cell lineage related genes such as TP63, Pax6, KRT14, and activation of Wnt, Notch, Hippo, and Hedgehog signaling pathways during the differentiation process. Tissue engineered epithelial cell sheet derived from hESCs showed stratified structure and normal corneal epithelial phenotype with presence of clonogenic progenitor cells. Eight weeks after grafting the cell sheet onto the ocular surface of LSCD rabbit model, a full-thickness continuous corneal epithelium developed to fully cover the damaged areas with normal limbal and corneal epithelial phenotype. CONCLUSION: The tissue engineered corneal epithelium generated from a clinical-grade hESCs may be feasible in the treatment of limbal stem cell deficiency.

Acute ocular hypertension disrupts barrier integrity and pump function in rat corneal endothelial cells.

Acute ocular hypertension (AOH) frequently compromises corneal endothelial cell (CEC) function in clinical practice. This type of stress induces corneal oedema and a decrease in the corneal endothelial cell density (ECD). The anterior chamber of the right eye of Sprague-Dawley rats was irrigated with Balanced Salt Solution (BSS) for two hours, and the left eye served as a control to determine the time-dependent effects of AOH on endothelial cell morphology and function. The average intraocular pressure (IOP) increased to 82.6 ± 2.3 mmHg (normal range: 10.2 ± 0.4 mmHg) during anterior irrigation. Very soon after initiating irrigation, corneal oedema became evident and the cornea exhibited a significant increase in permeability to FITC-dextran. The peripheral ECD was significantly reduced, and the morphology of CECs became irregular and multiform. The structures of the zonula occludens-1 (ZO-1) and F-actin were severely disrupted. In addtion, Na,K-ATPase exhibited a dispersed expression pattern. Two days after irrigation, obvious CEC proliferation was observed, the ECD recovered to a normal level, and F-actin was dispersed throughout the cytoplasm. Seven days later, the CEC structure and function were nearly normalized. Based on the results obtained using this model, an acute IOP crisis exerts transient deleterious effects on CEC structure and function in rats.