Efficacy of topical 0.05% cyclosporine A and 0.1% sodium hyaluronate in post-refractive surgery chronic dry eye patients with ocular pain.

BACKGROUND: The management of post-refractive surgery dry eye disease (DED) can be challenging in clinical practice, and patients usually show an incomplete response to traditional artificial tears, especially when it is complicated with ocular pain. Therefore, we aim to investigate the efficacy of combined topical 0.05% cyclosporine A and 0.1% sodium hyaluronate treatment in post-refractive surgery DED patients with ocular pain unresponsive to traditional artificial tears. METHODS: We enrolled 30 patients with post-refractive surgery DED with ocular pain who were unresponsive to traditional artificial tears. Topical 0.05% cyclosporine A and 0.1% sodium hyaluronate were used for 3 months. They were evaluated at baseline and 1 and 3 months for dry eye and ocular pain symptoms and objective parameters, including Numerical Rating Scale (NRS), Neuropathic Pain Symptom Inventory modified for the Eye (NPSI-Eye), tear break-up time (TBUT), Schirmer I test (SIt), corneal fluorescein staining (CFS), corneal sensitivity, and corneal nerve morphology. In addition, tear levels of inflammatory cytokines and neuropeptides were measured using the Luminex assay. RESULTS: After 3 months of treatment, patients showed a statistically significant improvement in the ocular surface disease index (OSDI), TBUT, SIt, CFS, and corneal sensitivity (all P < 0.01) using linear mixed models. As for ocular pain parameters, the NRS and NPSI-Eye scores were significantly reduced (both P < 0.05) and positively correlated with the OSDI and CFS scores. Additionally, tear IL-1ß, IL-6, and TNF-α levels were improved better than pre-treatment (P = 0.01, 0.03, 0.02, respectively). CONCLUSION: In patients with post-refractive surgery DED with ocular pain, combined topical 0.05% cyclosporine A and 0.1% sodium hyaluronate treatment improved tear film stability, dry eye discomfort, and ocular pain, effectively controlling ocular inflammation. TRIAL REGISTRATION: Registration number: NCT06043908.

Evaluation of left ventricular stiffness with echocardiography.

Half of patients with heart failure are presented with preserved ejection fraction (HFpEF). The pathophysiology of these patients is complex, but increased left ventricular (LV) stiffness has been proven to play a key role. However, the application of this parameter is limited due to the requirement for invasive catheterization for its measurement. With advances in ultrasound technology, significant progress has been made in the noninvasive assessment of LV chamber or myocardial stiffness using echocardiography. Therefore, this review aims to summarize the pathophysiological mechanisms, correlations with invasive LV stiffness constants, applications in different populations, as well as the limitations of echocardiography-derived indices for the assessment of both LV chamber and myocardial stiffness. Indices of LV chamber stiffness, such as the ratio of E/e' divided by left ventricular end-diastolic volume (E/e'/LVEDV), the ratio of E/SRe (early diastolic strain rates)/LVEDV, and diastolic pressure-volume quotient (DPVQ), are derived from the relationship between echocardiographic parameters of LV filling pressure (LVFP) and LV size. However, these methods are surrogate and lumped measurements, relying on E/e' or E/SRe for evaluating LVFP. The limitations of E/e' or E/SRe in the assessment of LVFP may contribute to the moderate correlation between E/e'/LVEDV or E/SRe/LVEDV and LV stiffness constants. Even the most validated measurement (DPVQ) is considered unreliable in individual patients. In comparison to E/e'/LVEDV and E/SRe/LVEDV, indices like time-velocity integral (TVI) measurements of pulmonary venous and transmitral flows may demonstrate better performance in assessing LV chamber stiffness, as evidenced by their higher correlation with LV stiffness constants. However, only one study has been conducted on the exploration and application of TVI in the literature, and the accuracy of assessing LV chamber stiffness remains to be confirmed. Regarding echocardiographic indices for LV myocardial stiffness evaluation, parameters such as epicardial movement index (EMI)/ diastolic wall strain (DWS), intrinsic velocity propagation of myocardial stretch (iVP), and shear wave imaging (SWI) have been proposed. While the alteration of DWS and its predictive value for adverse outcomes in various populations have been widely validated, it has been found that DWS may be better considered as an overall marker of cardiac function performance rather than pure myocardial stiffness. Although the effectiveness of iVP and SWI in assessing left ventricular myocardial stiffness has been demonstrated in animal models and clinical studies, both indices have their limitations. Overall, it seems that currently no echocardiography-derived indices can reliably and accurately assess LV stiffness, despite the development of several parameters. Therefore, a comprehensive evaluation of LV stiffness using all available parameters may be more accurate and enable earlier detection of alterations in LV stiffness.

Seeking information about waste-to-energy incineration projects: The role of objective knowledge and benefit perceptions in an extended PRISM.

Although waste-to-energy (WtE) incineration projects have boosted the economic effectiveness of the waste management system, locals frequently view them with suspicion, opposition, or even outright rejection because of potential environmental and health risks. In this study, by incorporating two additional variables, namely, objective knowledge and benefit perceptions, the planned risk information seeking model (PRISM) was extended in the context of the WtE incineration project. A total of 1726 respondents were interviewed in person. Results from the structural equation modeling indicated that the extended PRISM achieved excellent fits, suggesting the generalizability of the model in these contexts. Objective knowledge and subjective knowledge have a positive and direct impact on perceived knowledge insufficiency. The impact of benefit and risk perceptions on worry is the opposite, with risk perceptions having a greater direct effect on worry. However, compared with risk perceptions, the direct effect that benefit perceptions have on perceived knowledge insufficiency is stronger. Additionally, perceived knowledge insufficiency partially or completely mediates the relationships between all antecedents, except perceived seeking control, and information-seeking intention. This study suggests that risk managers carefully consider how to establish powerful communication tactics to shape these antecedents when introducing a "not in my backyard" project to the local community.

Impact of Topical 0.05% Cyclosporine A Eye Drops on Post-Femtosecond-Assisted Laser In Situ Keratomileusis Ocular Surface Recovery: A Randomized Clinical Trial.

OBJECTIVES: To investigate the effect of topical 0.05% cyclosporine A (CsA) eye drops as an adjunct to conventional therapy in maintaining post-femtosecond-assisted laser in situ keratomileusis (FS-LASIK) ocular surface stability. METHODS: Sixty-six patients (eyes) undergoing FS-LASIK were randomized into 2 groups: 33 patients (eyes) in group I (conventional treatment group) and 33 patients (eyes) in group II (CsA group). Conventional treatments include topical levofloxacin, fluorometholone, and artificial tears. Group II received topical 0.05% CsA eye drops twice daily for three months in addition to conventional treatment. Ocular Surface Disease Index (OSDI), numerical rating scale (NRS), tear break-up time (TBUT), Schirmer I test (SIt), corneal fluorescein staining (CFS), conjunctival lissamine green (LG) staining, corneal sensitivity, and corneal nerve morphology were measured. In addition, tear inflammatory cytokine levels were measured using the Luminex assay. Follow-up was performed preoperatively and 1 and 3 months postoperatively. RESULTS: In the CsA group, OSDI, TBUT, LG, corneal sensitivity, and corneal nerve fiber total branch density recovered better than in the conventional treatment group. As for tear inflammatory cytokines, interferon (INF) -γ, interleukin (IL)-10, and IL-6 levels were significantly higher in the conventional treatment group as compared with the CsA group. In addition, no significant differences in NRS, SIt, and CFS scores were observed between the two groups. CONCLUSION: In conclusion, 0.05% CsA eye drops is a useful adjunct to conventional treatment for restoring the ocular surface stability after corneal refractive surgery and is more potent in sustaining anti-inflammatory effects.

Comparison of mucin levels at the ocular surface of visual display terminal users with and without dry eye disease.

BACKGROUND: The long-term use of visual display terminals (VDT) is linked to an increased risk of dry eye disease (DED). Numerous studies have indicated that ocular mucins play a vital role in the pathogenesis of DED. Therefore, we aimed to evaluate (1) whether mRNA levels of membrane-associated mucins (MAMs), including MUC1, MUC4, MUC16, and MUC20, as well as MUC5AC are altered in conjunctival cells of VDT users with and without DED and (2) the relationship between mucin levels and subjective and objective tests of DED in VDT users. METHODS: Seventy-nine VDT users were enrolled and divided into DED (n = 53) and control (n = 26) groups. All participants were evaluated for parameters of DED using the Ocular Surface Disease Index (OSDI) questionnaire, tear breakup time (TBUT), corneal fluorescein staining (CFS), lissamine green (LG) staining, and tear meniscus height (TMH). Based on the conjunctival impression cytology (CIC) method, differences in MUC1, MUC4, MUC16, MUC20, and MUC5AC mRNA expression levels were observed between the DED and control groups, and between symptomatic and asymptomatic participants. RESULTS: The DED group showed significantly decreased MUC1, MUC16, and MUC20 expressions (all P < 0.05) compared to the control group. In addition, these mucin levels were lower in subjects with frequent ocular symptoms (foreign body sensation, blurred vision and painful or sore eyes) than in asymptomatic participants (all P < 0.05). Correlation analysis revealed that MUC1, MUC16, and MUC20 levels in VDT users were positively correlated with TBUT or TMH, or both. However, no significant relationship was found between MUC4 and MUC5AC levels and the DED parameters. CONCLUSION: VDT users with an increased frequency of ocular discomfort or a diagnosis of DED had a decreased MUC1, MUC16 and MUC20 mRNA expression in their conjunctival cells. MAMs deficiency in the conjunctival epithelium may be one of the mechanisms leading to tear film instability and DED in VDT users.

Evaluation of biometric indicators of anterior segment parameters after ICL implantation by swept-source optical coherence tomography.

BACKGROUND: To evaluate anterior segment structural alterations after implantable collamer lens (ICL) implantation in myopic patients using swept-source quantitative optical coherence tomography (SS-OCT). METHODS: This prospective study included 47 eyes in 24 patients with preoperative spherical equivalent ≥ -3.00 D. Patients underwent ICL implantation at Department of Ophthalmology, Peking University Third Hospital, from May 2021 to December 2022. SS-OCT was used to measure anterior chamber width (ACW), angle opening distance (AOD), angle recess area (ARA), trabecular-iris area (TISA), trabecular-iris angle (TIA), iridotrabecular contact (ITC) area, and ITC Index before ICL implantation surgery and at 1 month follow-up. The correlations among the ITC index, vault, and angle parameters were analysed. Receiver operating characteristic (ROC) analysis was used to explore the ability of the vault to identify eyes with suspected angle-closure. RESULTS: At one month following ICL implantation, the ITC area was 0.396 ± 0.37 mm2, and the ITC index is 8.143 ± 5.439%. All angle parameters, except ACW, showed a statistically significant reduction on SS-OCT (P < 0.05). Mean AOD500, AOD750, ARA500, ARA750, TISA500, TISA750, TIA500, and TIA750 values at one month postoperatively decreased by 60.0%, 60.4%, 58.1%, 59.2%, 57.3%, 58.7%, 48.8%, and 50.7%, respectively. The vault was positively correlated with the ITC index and percent change in anterior chamber angle parameters. A vault of > 0.659 mm was found to be optimal for angle-closure suspect with a sensitivity of 85.2% and a specificity of 53.9%. CONCLUSIONS: Anterior chamber angle parameters decreased one month after ICL implantation, and their percentage changes and ITC index correlated with the vault. When the vault is larger than 0.659 mm, it is necessary to be alert to possible closed angle suspicion.

Impaired Cardiomyocyte Maturation Leading to DCM: A Case Report and Literature Review.

Background: The maturation of cardiomyocytes is a rapidly evolving area of research within the field of cardiovascular medicine. Understanding the molecular mechanisms underlying cardiomyocyte maturation is essential to advancing our knowledge of the underlying causes of cardiovascular disease. Impaired maturation can lead to the development of cardiomyopathy, particularly dilated cardiomyopathy (DCM). Recent studies have confirmed the involvement of the ACTN2 and RYR2 genes in the maturation process, facilitating the functional maturation of the sarcomere and calcium handling. Defective sarcomere and electrophysiological maturation have been linked to severe forms of cardiomyopathy. This report presents a rare case of DCM with myocardial non-compaction, probably resulting from allelic collapse of both the ACTN2 and RYR2 genes. Case Presentation: The proband in this case was a four-year-old male child who presented with a recurrent and aggressive reduction in activity tolerance, decreased ingestion volume, and profuse sweating. Electrocardiography revealed significant ST-T segment depression (II, III, aVF V3-V6 ST segment depression >0.05 mV with inverted T-waves). Echocardiography showed an enlarged left ventricle and marked myocardial non-compaction. Cardiac magnetic resonance imaging revealed increased left ventricular trabeculae, an enlarged left ventricle, and a reduced ejection fraction. Whole exome sequencing revealed a restricted genomic depletion in the 1q43 region (chr1:236,686,454-237,833,988/Hg38), encompassing the coding genes ACTN2, MTR, and RYR2. The identified variant resulted in heterozygous variations in these three genes, with the ACTN2 g.236,686,454-236,764,631_del and RYR2 g.237,402,134-237,833,988_del variants being the dominant contributors to the induction of cardiomyopathy. The patient was finally diagnosed with DCM and left ventricular myocardial non-compaction. Conclusions: This study reports a rare case of DCM with myocardial non-compaction caused by the allelic collapse of the ACTN2 and RYR2 genes. This case provides the first human validation of the critical role of cardiomyocyte maturation in maintaining cardiac function and stability and confirms the key findings of previous experimental research conducted by our group. This report emphasizes the connection between genes involved in regulating the maturation of cardiomyocytes and the development of cardiomyopathy.

Assessing the efficacy of four methods established by four parameters in ICL size selection and relevant influencing factors: a prospective cohort study.

PURPOSE: To compare the efficacy and relevant influencing factors of four ICL size selection methods established by four different parameters. METHODS: This prospective study included 60 patients (120 eyes) who underwent bilateral ICL implantation. Patients were equally divided into four groups, and each group used the Parkhurst nomogram based on sulcus-to-sulcus (STS), the manufacturer's Online Calculation & Ordering System (OCOS) nomogram based on white-to-white (WTW), the KS formula based on angle-to-angle (ATA) and the NK formula based on anterior chamber width (ACW) to determine the ICL size. Recorded the vault one month after operation and compared the consistency between STS and WTW, ATA and ACW and their effects on the vault of different groups. RESULTS: The Parkhurst nomogram, OCOS nomogram, KS formula and NK formula determined 86.7%, 70.0%, 83.3% and 66.7% of properly sized ICL, respectively. STS and ATA were correlated (P < 0.05). The mean difference between the STS and WTW, ATA and ACW was -0.37 ± 0.62 mm, -0.42 ± 0.53 mm and -0.44 ± 0.52 mm, respectively. The vault in the OCOS group was negatively correlated with △STS-WTW, and the vault in the NK group was negatively correlated with △STS-WTW, △STS-ATA and △STA-ACW. The vault in the Parkhurst group and KS group was not affected by anterior segment biometry variables. CONCLUSION: ATA can be served as an alternative parameter to STS, and STS-based Parkhurst nomogram and ATA-based KS formula determined the most appropriate ICL size. When using OCOS nomogram and NK formula to select ICL size, postoperative abnormal vault was associated with a larger difference between STS and other anterior segment parameters.

Binocular visual function after unilateral versus bilateral implantation of segmented refractive multifocal intraocular lenses: a pilot study.

PURPOSE: To evaluate binocular visual function after unilateral and bilateral implantation of segmented refractive multifocal intraocular lenses (MIOLs). METHODS: This prospective comparative pilot study included patients who underwent SBL-3 (Lenstec; + 3.00 D) implantation at Peking University Third Hospital. Patients were divided into two groups (monocular or binocular surgery). Thirty-two patients with emmetropic presbyopic contralateral eyes and 49 patients with bilateral SBL-3 implantation within a week between eyes were included in the unilateral SBL-3 and bilateral groups, respectively. At 3-month follow-up, the main outcomes were binocular uncorrected distant, intermediate, and near visual acuity (UDVA, UIVA, and UNVA). Secondary outcomes included binocular best-corrected visual acuity at all distances, defocus curve, contrast sensitivity, photic phenomena, spectacle independence, patient satisfaction, and National Eye Institute Visual Function Questionnaire (NEI-VFQ-25) score. The essential perceptual phenomena constituting binocular vision, simultaneous perception, fusion, and stereopsis were also evaluated. RESULTS: Both groups showed similar binocular UDVA and UIVA, but UNVA was significantly better in the bilateral group (0.07 ± 0.07 versus 0.12 ± 0.07, P = 0.008). Better binocular defocus curve at intermediate to near focal points, higher percentage of spectacle independence at near distance, and NEI-VFQ-25 near activity scores were observed in the bilateral group. No significant differences in contrast sensitivity, photic phenomena, overall satisfaction, other NEI-VFQ-25 subscales, fusional amplitude, and stereoacuity were found between groups. CONCLUSION: Unilateral implantation of segmented refractive MIOL provided desirable distant visual acuity and high patient satisfaction, but inferior intermediate and near visual outcomes compared with bilateral implantation.

Evaluation of Ocular Surface Characteristics in Dry Eye Disease With and Without Soft Contact Lens Wear: A Comparative Study.

OBJECTIVES: To investigate ocular surface alterations and in vivo confocal microscopic characteristics of the cornea in dry eye disease (DED) with contact lens wear (CLW). METHODS: Sixty participants were divided into three groups: DED with CLW (n=20), DED without CLW (n=20), and normal control (n=20). Ocular surface parameters were evaluated. Basal tears and in vivo confocal microscopy images of the cornea were collected. Multiplex bead analysis was used to assess interleukin (IL)-6, IL-1ß, tumor necrosis factor (TNF)-α, nerve growth factor (NGF), and substance P (SP) in tears. Nerve morphology and dendritic cell density in corneal subbasal nerve images were calculated. RESULTS: The DED with CLW group showed significantly higher ocular surface staining scores ( P =0.022) and higher levels of IL-1ß, NGF, and SP in tears ( P =0.014, P =0.004 and P =0.025) than the DED without CLW group. Corneal dendritic cell density in the DED with CLW group was significantly higher than that in the normal controls ( P =0.001) and DED without CLW group ( P =0.043). Tear cytokine levels of IL-1ß, NGF, and SP were correlated with ocular surface parameters in the DED with CLW group. Moreover, the years of CLW were positively correlated with corneal dendritic cell density (r=0.527, P =0.017) and negatively correlated with corneal nerve density (r=-0.511, P =0.021). CONCLUSIONS: Patients with DED with CLW showed greater epithelial damage, elevated inflammatory cytokines and neuromediators in tears, and higher corneal dendritic cell density than patients with DED without CLW. The immune and nervous systems may be involved in contact lens-related DED.

Predictive value of coagulation profiles for both initial and repeated immunoglobulin resistance in Kawasaki disease: A prospective cohort study.

BACKGROUND: Intravenous immunoglobulin (IVIG) resistance prediction remains substantial in Kawasaki disease (KD), with limited data on the predictive value of coagulation profile for IVIG resistance, particularly for repeated IVIG resistance. Therefore, the aim of our study was to testify the predictive validity of coagulation profile for both initial IVIG resistance and repeated IVIG resistance in KD. METHODS: A total of 385 KD patients were prospectively recruited between April 2015 and May 2019. Coagulation and other profiles were evaluated between the IVIG-responsive and IVIG-resistant groups. Multivariate logistic regression analysis was applied to determine the association between coagulation profiles and IVIG resistance. ROC curves analysis was further performed to assess the validity of coagulation profiles in predicting both initial IVIG resistance and repeated IVIG resistance. RESULTS: Prothrombin time (PT), activated partial thromboplastin time (APTT), international normalized ratio (INR), fibrinogen degradation products (FDPs), and D-dimer were significantly increased in the initial IVIG-resistant group with antithrombin III (ATIII) and thrombin time (TT) significantly reduced. Meanwhile, ATIII was declined markedly in repeated IVIG-resistant patients. Multivariate logistic regression analysis showed that PT, APTT, D-dimer, and ATIII were independent risk factors for predicting initial IVIG resistance and ATIII for predicting repeated IVIG-resistant patients with KD. PT, APTT, D-dimer, and ATIII cutoff values of 13.95 s, 41.15 s, 1.48 mg/L, and 89.5% yielded sensitivities of 73%, 32%, 71%, and 81%, and specificities of 55%, 88%, 62%, and 51% for predicting initial IVIG resistance, respectively. The cutoff value of ATIII for predicting repeated IVIG resistance was 68.5%, with sensitivity of 71% and specificity of 55%. CONCLUSIONS: KD patients who have hypercoagulation during the acute phase might be at higher risk of developing IVIG resistance.

A severe coronavirus disease 2019 patient with high-risk predisposing factors died from massive gastrointestinal bleeding: a case report.

BACKGROUND: SARS-CoV-2 is highly infectious and has been a significant public health threat. Despite typical manifestations of illness are dominated by respiratory symptom, some patients have concurrent gastrointestinal manifestations, including nausea, diarrhea, and vomiting. Massive gastrointestinal bleeding, however, has rarely been reported. CASE PRESENTATION: We herein described a case of severe SARS-CoV-2 infected patient with several risk factors for poor prognosis, including male, hypertension, old age, mixed bacterial infection and multilobular infiltration on radiological imaging. After improvement of respiratory status, the onset of gastrointestinal bleeding occurred, probably resulting from direct viral invasion as evidenced by the positive findings for SARS-CoV-2 in the repeat stool specimens. Although aggressive resuscitation was administered, hematochezia was uncontrolled. The patient rapidly deteriorated, suffered from cardiac arrest, and expired. CONCLUSIONS: Digestive symptoms could be severe in SARS-CoV-2 infected patients, especially for the high-risk individuals with predisposing conditions. A more thorough protocol for preventing cross-infection through faecal-oral transmission should be implemented in the process of patient care and infection control.

The effect of maternal exposure to di-(2-ethylhexyl)-phthalate on fetal cardiac development in mice.

Accumulating evidence has suggested a link between maternal di-(2-ethylhexyl)-phthalate (DEHP) exposure and various developmental abnormalities. However, the evidence regarding the effect of maternal DEHP exposure on fetal cardiac development is scarce. The present study aimed to determine the effect of maternal DEHP exposure on fetal cardiac development in mice and explore the possible involved mechanism preliminarily. The C57BL mice were randomly divided into four groups: the vehicle group (corn oil, n = 10), 250 mg kg-1 DEHP group (n = 15), 500 mg kg-1 DEHP group (n = 20) and 1 g kg-1 DEHP group (n = 20). Pregnant dams in different group received respective intervention by gavage once daily from embryonic day (E)6.5 to E14.5. Maternal weights were monitored every day and samples were collected at E15.5. Hematoxylin and eosin staining was used to examine fetal cardiac malformations. Real-time quantitative polymerase chain reaction and western blot were applied to detect peroxisome proliferator-activated receptor (PPAR)α/PPARγ/Nkx2.5/Gata4/Tbx5/Mef2c/Chf1 mRNA and protein expression, respectively. Maternal DEHP exposure significantly decreased maternal body weight, fetal weight and placental weight, and remarkably elevated fetal cardiac malformations rate. The phenotypes of cardiac anomalies mainly include septal defects, ventricular myocardium noncompaction and cardiac hypoplasia. Higher doses DEHP (500 mg kg-1 and 1 g kg-1 ) could significantly decreased fetal cardiac Gata4/Mef2c/Chf1 expression, while PPARγ expression was upregulated. Maternal exposure to higher doses of DEHP could result in fetal cardiac development malformations in mice and it might have resulted from the inhibition of cardiac GATA4/Mef2c/Chf1 expression via PPARγ activation.

Circadian rhythm genes mediate fenvalerate-induced inhibition of testosterone synthesis in mouse Leydig cells.

Fenvalerate (Fen), a widely used pesticide, is known to impair male reproductive functions by mechanisms that remain to be elucidated. Recent studies indicated that circadian clock genes may play an important role in successful male reproduction. The aim of this study was to determine the effects of Fen on circadian clock genes involved in the biosynthesis of testosterone using TM3 cells derived from mouse Leydig cells. Data demonstrated that the circadian rhythm of testosterone synthesis in TM3 cells was disturbed following Fen treatment as evidenced by changes in the circadian rhythmicity of core clock genes (Bmal1, Rev-erbα, Rorα). Further, the observed altered rhythms were accompanied by increased intracellular Ca2+ levels and modified steroidogenic acute regulatory (StAR) mRNA expression. Thus, data suggested that Fen inhibits testosterone synthesis via pathways involving intracellular Ca2+ and clock genes (Bmal1, Rev-Erbα, Rorα) as well as StAR mRNA expression in TM3 cells.

[Effect of histone acetylation/deacetylation imbalances on key gene of planar cell polarity pathway].

OBJECTIVE: To investigate the effect of histone acetylation/deacetylation imbalances on embryonic hearts of mice and its effect on key genes of planar cell polarity (PCP) pathway-Vangl2, Scrib and Rac1 in H9C2 cells. METHODS: Forty pregnant C57/B6 mice were randomly assigned into three groups: blank group (n=10), vehicle group (n=10), and valproic acid (VPA)-treated group (n=20). In the VPA-treated group, VPA, a histone deacetylase (HDAC) inhibitor, was administered to each individual dam intraperitoneally at a single dose of 700 mg/kg on embryonic day 10.5 (E10.5). The vehicle and blank groups received equivalent saline or no interventions, respectively. Dams were sacrificed on E15.5, and death rates of embryos were evaluated. Subsequently, embryonic hearts of survival fetus were removed to observe cardiac abnormalities by hematoxylin-eosin (HE) staining. H9C2 cells were cultured and allotted to the blank, vehicle, and VPA-treated groups: the VPA treated group received VPA exposure at concentrations of 2.0, 4.0 and 8.0 mmol/L; the vehicle and blank groups received equivalent saline or no interventions, respectively. HDAC1-3 as well as Vangl2, Scrib and Rac1 mRNA and protein expression levels were determined by quantitative real-time PCR and Western blot, respectively. The total HDAC activity was analyzed by colorimetric assay. RESULTS: The fetus mortality rate after VPA treatment was 31.7%, with a significantly higher rate of cardiac abnormalities in comparison with the controls (P<0.05). In comparison with the blank and vehicle groups, HDAC1 mRNA was significantly increased at various concentrations of VPA treatment at all time points of exposure (P<0.05), together with a reduction of protein level after 48 and 72 hours of exposure (P<0.05). The inhibition of HDAC2 mRNA after various concentrations of VPA incubation was pronounced at 24 hours of exposure (P<0.05), while the protein levels were reduced at all time points (P<0.05). HDAC3 mRNA was prominently induced by VPA (4.0 and 8.0 mmol/L) at all time points of treatment (P<0.05). In contrast, the protein level was inhibited after VPA treatment (P<0.05). In comparison with the blank and vehicle groups, Vangl2 mRNA as well as Scrib mRNA/protein expression levels were markedly reduced after 48 and 72 hours of VPA treatment (P<0.05), together with a reduction of protein level in Vangl2 at 72 hours (P<0.05). Compared with the blank and vehicle groups, a significant repression in the total HDAC activity was observed in the VPA-treated group at concentrations of 4.0 and 8.0 mmol/L after 24 hours of treatment (P<0.05), and the effect persisted up to 48 and 72 hours, exhibiting pronounced inhibition at all concentrations (P<0.05). CONCLUSIONS: VPA might result in acetylation/deacetylation imbalances by inhibiting HDAC1-3 protein expression and total HDAC activity, leading to the down-regulation of mRNA and protein expression of Vangl2 and Scrib. This could be one of the mechanisms contributing to congenital heart disease.

Melatonin attenuates neurogenic pulmonary edema via the regulation of inflammation and apoptosis after subarachnoid hemorrhage in rats.

Neurogenic pulmonary edema (NPE) is a serious non-neurological complication that can occur after a subarachnoid hemorrhage (SAH) and is associated with decreased survival and a poor neurological outcome. Melatonin is a strong antioxidant that has beneficial effects against SAH in rats, including reduced mortality and reduced neurological deficits. The molecular mechanisms underlying these clinical effects in the SAH model, however, have not been clearly identified. This study was undertaken to determine the influence of melatonin on SAH-induced NPE and the potential mechanism of these effects using the filament perforation model of SAH in male Sprague Dawley rats. Either melatonin (150 mg/kg) or a vehicle was given via an intraperitoneal injection 2 hr after an SAH induction. Lung samples were extracted 24 hr after SAH. The results show that the melatonin treatment attenuated SAH-induced NPE by preventing alveolar-capillary barrier dysfunctions via inhibiting the disruption of tight junction proteins (ZO-1 and occludin). Moreover, the treatment downregulated the levels of mature interleukin (IL) -1ß, myeloperoxidase (MPO), and matrix metallopeptidase (MMP) 9 expression/activation, which were increased in the lung; also, melatonin treatment improved neurological deficits. Furthermore, the melatonin treatment markedly reduced caspase-3 activity and the number of TUNEL-positive cells in the lung. Taken together, these findings show that administration of melatonin attenuates NPE by preventing alveolar-capillary barrier dysfunctions via repressing the inflammatory response and by anti-apoptosis effects after SAH.

A rare homozygous variant of CHKB induced severe cardiomyopathy and a cardiac conduction disorder: a case report.

Background: The CHKB (choline kinase beta) gene plays a crucial role in regulating mitochondrial function and choline metabolism. Mutations in CHKB lead to conditions such as megaconial congenital muscular dystrophy (MCMD), characterized by enlarged mitochondria and impaired mitochondrial function, inducing various clinical features in neurological and cardiac performance. Herein, we report a rare case presenting with dilated cardiomyopathy as the dominant feature with a homozygous nonsense variant of CHKB, and the related therapeutic strategy. Case presentation: The proband, a 13-year-old male, presented with a complex clinical profile characterized by mild intellectual disability and severe cardiac impairment, including reduced activity tolerance, suspected acute heart failure, significant cardiac enlargement, a left anterior fascicular block, and a complete right bundle branch block. Whole exome sequencing (WES) identified a homozygous nonsense variant, c.598delC (p.Q200Rfs*11) of the CHKB gene, that resulted in disease caused by amino acid sequence changes, a truncated protein, and splice site changes, as demonstrated by MutationTaster analysis. The protein structure of CHKB was built and named AF-Q9Y259-F1. The residue around 200 amino acid sites changed in CHKB p.Q200Rfs*11 with unaltered hydrogen bonds which indicated the pathogenicity of the variant mainly originated from a truncated protein induced by the nonsense mutation. The heart blocks in the proband were considered to be associated with choline metabolic impairment, and thus cardiac resynchronization therapy would benefit the patient. Furthermore, the missense homozygous or compound heterozygous variants of CHKB, as well as the combined compound heterozygous missense and nonsense variants of CHKB, usually lead to neurological impairments and muscular weakness. Conclusion: This study expands the spectrum of CHKB mutations and provides essential information for the genotype-phenotype map of a nonsense variant of the gene. It is important to confirm a differential diagnosis among such patients using WES analyses. Regular cardiac and musculoskeletal monitoring is recommended for MCMD patients. Patients with a CHKB deficiency presenting with heart blocks could benefit from the administration of cardiac resynchronization therapy. This therapeutic approach might improve cardiac function and conduction in patients with CHKB-related cardiomyopathies.