While large angle kappa (chord µ) and angle alpha (chord α) have been reported to adversely affect visual quality with diffractive multifocal intraocular lenses (MIOLs), their impact on the postoperative visual outcomes of nondiffractive extended depth of focus (EDOF) IOLs remains unclear. This study aimed to evaluate the influence of postoperative angle kappa and angle alpha on visual and optical outcomes following implantation of a nondiffractive EDOF IOL. This prospective observational study included 115 cataract patients (115 eyes) who underwent implantation of the AcrySof® IQ Vivity® nondiffractive wavefront-shaping EDOF IOL. At 3 months postoperatively, photopic and mesopic angle kappa and angle alpha were measured using the OPD-Scan III. Visual outcomes included uncorrected visual acuity at different distances, objective optical quality parameters, contrast threshold, and patient-reported subjective visual disturbance scores. We performed partial correlation analyses to examine the relationships of angle kappa and angle alpha with postoperative visual outcomes, while controlling for potential confounding variables. Subgroup analyses based on clinically relevant offset thresholds were additionally conducted. At 3 months postoperatively, most postoperative angle kappa and angle alpha values were within physiological ranges, with only a small number of eyes having values ≥ 0.5 mm (photopic angle kappa: n = 7; mesopic angle kappa: n = 10; angle alpha: n = 29). Partial correlation analyses demonstrated no significant associations between photopic or mesopic angle kappa and postoperative visual acuity, objective optical quality metrics, or subjective visual disturbance scores (all P > 0.05). Angle alpha showed weak but statistically significant positive correlations with uncorrected distance visual acuity and contrast threshold (P < 0.05), indicating slightly better visual performance with larger angle alpha values. No significant differences in visual outcomes were observed among angle kappa and angle alpha subgroups. Within the observed postoperative range, angle kappa and angle alpha were not associated with clinically meaningful deterioration in visual performance following implantation of a nondiffractive EDOF IOL. These findings suggest that wavefront-shaping EDOF designs may be tolerant to moderate angular offsets in routine clinical practice.

CUT-like homeobox 1 (CUX1) is a transcription factor with dual roles in tumorigenesis. Among its splice variants, the Golgi-localized cut alternative spliced product (CASP) lacks DNA-binding domains, and its functional significance has remained largely unexplored. In this study, we identify CASP as a potential oncoprotein that is specifically upregulated in colorectal adenomas and carcinomas. Genetic silencing of CASP suppressed the proliferation and migration of colorectal cancer (CRC) cells, while its overexpression enhanced tumor growth and metastatic potential both in vitro and in vivo. Mechanistically, CASP interacts with the E3 ubiquitin ligase TRIM21 and promotes its ubiquitination and proteasomal degradation, leading to subsequent activation of the mitogen-activated protein kinase (MAPK) signaling pathway. Clinically, CASP expression was correlated with mismatch repair (MMR)/microsatellite instability (MSI) status and TP53 mutational profiles. These findings implicate CASP in CRC progression and support its potential as a prognostic biomarker and therapeutic target by disrupting CASP-driven signaling.

BACKGROUND:Dry age-related macular degeneration (AMD) is driven largely by retinal pigment epithelium (RPE) cell injury. Kinsenoside (KN) exhibits protective activity on RPE cells, but the direct effect on dry AMD and underlying molecular mechanisms remain unclear. OBJECTIVE:This study aimed to investigate the protective effects of KN against RPE cell injury and dysfunction in dry AMD and to elucidate its regulatory mechanisms. METHODS:An A2E- and blue light-induced ARPE-19 cell injury model and a blue light-induced AMD-like retinal injury mouse model were established. RPE cytotoxicity, apoptosis, senescence, inflammation, and melanogenesis-associated marker expression were evaluated. MAPK signaling was analyzed by Western blotting, and mechanistic analyses were performed using a JNK inhibitor, molecular docking analysis, an in vitro JNK enzymatic activity assay, exosome characterization, and exosome functional assays. RESULTS:KN treatment significantly attenuated cytotoxicity, apoptosis, cellular senescence, inflammatory responses, and melanogenesis-associated marker alterations in AMD-like ARPE-19 cells. In vivo, KN reduced drusen-like deposition, improved retinal structural integrity, and partially restored outer nuclear layer thickness while suppressing senescence and inflammation. Mechanistically, KN preferentially attenuated pathological JNK activation under the examined conditions and reduced JNK enzymatic activity in vitro. Pharmacological JNK inhibition phenocopied the protective effects of KN with no additive benefit, indicating pathway convergence. Exosome inhibition, transfer, and depletion experiments supported the contribution of exosome-associated signaling to KN-mediated cytoprotection. CONCLUSION:These findings suggest that KN alleviates RPE cell injury under AMD-like stress, at least partly through exosome-associated modulation of JNK/MAPK signaling.

Biologics have emerged as frontline therapies for uveitis due to their targeted immunomodulatory effects. However, conventional monoclonal antibodies remain confined to single-target engagement. In addition, intravitreal injections carry risks of complications associated with repeated administration, whereas noninvasive delivery is hindered by compromised bioavailability across the complex ocular barriers. Together, these limitations restrict the effectiveness of current therapeutic approaches. Herein, bioinformatic screening identified VEGF and tumor necrosis factor alpha (TNF-α) as core pathogenic mediators in uveitis. Subsequently, we engineered a VEGF/TNF-α bispecific antibody (V5-3) and assembled it with a dithiolane molecule (DM) into nanoparticles (V5-3-NPs), thereby creating a noninvasive topical nanoformulation for uveitis therapy. Compared with V5-3, V5-3-NPs significantly enhanced cellular uptake, inhibited microglial proliferation and migration, and reduced the release of pro-inflammatory cytokines. In both in vitro and in vivo models, the synergistic bispecific targeting of V5-3-NPs enhanced anti-inflammatory and anti-angiogenic efficacy, thereby effectively alleviating clinical manifestations and histopathological damage. The negligible toxicity observed across models confirmed its safety profile and supported clinical feasibility. This work tackles key unmet needs in uveitis therapy by overcoming inefficient drug delivery and single-target constraints. By integrating bispecific targeting with nanocarrier-based delivery, it offers a distinct technological advance and a clinically viable strategy for uveitis treatment.

Intraocular injection of anti-vascular endothelial growth factor (VEGF) antibodies is the first-line treatment for ocular neovascular diseases. However, the invasive nature of this administration method often reduces patient compliance and negatively affects treatment outcomes. Noninvasive formulations of anti-VEGF antibody are urgently needed, but their development remains challenging due to the complex ocular barriers. This study identified an anti-VEGF single-domain antibody (sdVE01) that is three times smaller than the commercially available ranibizumab, yet retains a comparable anti-angiogenic effect to the heavy-chain region of ranibizumab (VHHL). Additionally, four dithiolane molecules (DM) were designed to construct DM-based antibody nanoformulations, which effectively penetrate both the anterior and posterior segments of the eye. Upon eyedrop administration, DM-based antibody nanoformulations significantly inhibited the VEGF pathway and reduced neovascularization in a corneal alkali-burn rat model. Notably, the therapeutic effects of the antibody eyedrops were comparable to those of ranibizumab administered via subconjunctival injection. Overall, the dithiolane-based antibody eyedrops represent a promising noninvasive strategy for treating ocular neovascularization diseases.

Up to 50% of patients with metastatic microsatellite instability-high (MSI-H) colorectal cancer (CRC) are resistant to immunotherapy and experience progression or recurrence after treatment. We integrated the genomic, epigenomic, transcriptomic, and proteomic data for 99 patients in a Chinese MSI-H CRC cohort. Proteomic profiling of primary tumors clearly classified MSI-H tumors into 2 subtypes. We found that the 2 subtypes have different mutational signatures, enriched pathways, gene fusion networks, and clinical outcomes. Notably, NCAM1 could serve as a potential biomarker for checkpoint inhibitor response in MSI-H CRC. Thus, there is an urgent need to stratify the MSI-H group into different subtypes and adopt more targeted therapies to prolong patient survival.

We previously discovered that HOXC6 was the most significantly upregulated gene in right-sided colon cancer compared to left-sided colon cancer according to our previous study; however, the role of HOXC6 in microsatellite instability-high (MSI-H) tumors remains poorly understood. Here, multiple public datasets, and in-house cohorts were used to analyze the differential expression and prognostic role of HOXC6 in colorectal cancer (CRC). Immunohistochemistry and immunofluorescence were performed to evaluate the correlation between HOXC6 expression and M2 macrophage infiltration. CCK8 and Transwell assays were used to evaluate the proliferation and migration of tumor cells in vitro. BALB/c nude mice were utilized to construct a humanized immune system model to evaluate the efficacy of ruxolitinib in vivo. We found that HOXC6 was overexpressed in MSI-H CRC and associated with a poor prognosis. Upregulation of CCL2 by HOXC6 increased M2 macrophage infiltration. IL6 secreted by M2 macrophages induced the epithelial-mesenchymal transition of tumor cells by upregulating HOXC6. M2 macrophages promoted effector T cell exhaustion by downregulating 4-1BB. Thus, inhibition of the IL6/JAK pathway in M2 macrophages restored 4-1BB expression and T-cell cytotoxicity offering a promising therapeutic target for the treatment of HOXC6-overexpressing MSI-H CRC.

OBJECTIVE:To evaluate the clinical efficacy of an modified surgical technique using scleral tunneling and Tenon's capsule for implantation of the Ahmed Glaucoma Drainage Valve (AGV) in refractory glaucoma. METHODS:This research involved the retrospective collection of data from 78 patients diagnosed with refractory glaucoma who underwent implantation of the AGV using a modified technique combining scleral tunneling with autologous Tenon's capsule. The analysis focused on preoperative and 60-month postoperative indices, which included the surgical technique employed, intraocular pressure (IOP) measurements, the utilization of antiglaucoma medications, rates of surgical success, and the incidence of postoperative complications. RESULTS:The findings suggest that the modified surgical technique exhibited an accuracy in the placement of implants. It also achieved remarkable postoperative control of IOP. The average IOP of the enrolled patients decreased from a preoperative maximum of 44.5 mmHg to 18.62 mmHg at 60 months postoperatively. Additionally, the usage of antiglaucoma medications decreased from an average of 2.88 preoperative medications to 1.76 medications at 60 months postoperatively. The rates of surgical success were recorded at 85.90% at 12 months, 82.54% at 24 months, 76.60% at 36 months, 71.88% at 48 months, and 66.67% at 60 months following the surgical procedure. It is noteworthy that no severe complications, including drain valve exposure and endophthalmitis, were reported. CONCLUSIONS:The modified AGV implantation technique effectively regulated IOP, reduced medication dependence, and minimized severe complications. However, limitations such as small sample size and lack of a control group necessitate further large-scale trials to confirm its efficacy.

Background/aim To investigate the independent relationships of visual impairment (VI) and Subjective cognitive complaints (SCC) with physical function impairment (PFI) and the interaction effect between VI and SCC on PFI in American older adults. Methods The data of this cross-sectional study was obtained from the 2005–2008 National Health and Examination Survey (NHANES) conducted in the United States. The VI criterion included both subjective self-reported eyesight conditions and objective visual acuity test results. The self-reported questionnaires were utilized to determine PFI and SCC. According to the survey design of NHANS, original data were weighted to produce nationally representative estimates. Both the unweighted original data and weighted estimates underwent analysis. Crude and adjusted logistic models were employed to assess the pairwise associations among VI, SCC, and PFI. To assess the interactive effect, measures such as the relative excess risk due to interaction (RERI), attributable proportion due to interaction (AP), and synergy index (S) were calculated. Results A total of 2,710 subjects (weighted n = 38,966,687) aged 60 years or older were included. Compared with subjects without subjective visual impairment (SVI), those with SVI had a significant positive association with PFI [weighted OR (95%CI): 3.11 (2.25, 4.31)]. After multi-variable adjusting, the relationship remained significant [weighted OR (95%CI): 1.90 (1.32, 2.72)]. Similarly, those with objective visual impairment (OVI) were positively associated with the risk of PFI in the crude model [weighted OR (95%CI): 2.35 (1.53, 3.61)] and adjusted model [weighted OR (95%CI): 1.84 (1.07, 3.17)]. Moreover, we found the association of SCC with an increased risk of FPI [crude weighted OR (95%CI): 5.02 (3.40, 7.40); adjusted weighted OR (95%CI): 3.29 (2.01, 5.38)]. Ultimately, the additive interaction showed there was a significant positive interaction term between SVI and SCC on PFI, while OVI and SCC did not. Conclusion Both VI and SCC were significantly associated with PFI in elder adults. Besides, there was a significant synergistic interaction between SVI and SCC on PFI, which indicated the improvement of SVI and SCC may be beneficial for the prevention of PFI. For the elderly, especially those with multiple disabilities, comprehensive and targeted approaches are imperative to foster their overall well-being and health.

Conjunctival fi brosis is a common postoperative complication of glaucoma fi ltration surgery, resulting in uncontrolled intraocular pressure and surgery failure. Therefore, there is an urgent need to understand the molecular mechanisms underlying conjunctival fi brosis and to explore novel pharmacologic anti-fibrosis fi brosis therapies for glaucoma fi ltration surgery. Herein, the 4-dimensional data-independent acquisition (4DDIA) quantitative proteomic results, coupled with experimental data, revealed the activation of the Wnt/ (3-catenin pathway in transforming growth factor (TGF)-(31-induced human conjunctival fi broblasts (HConFs). Treatment with ICG-001, a Wnt/(3-catenin inhibitor, effectively inhibited cell proliferation and migration in TGF(31-treated HConFs. ICG-001 treatment alleviated the increased generation of extracellular matrix proteins induced by TGF(31. In addition, ICG-001 reduced the expression level of a smooth muscle actin (a-SMA) and inhibited cell contractility in TGF(31-treated HConFs. Proteomics data further suggested that aB-crystallin (CRYAB) was a downstream target of Wnt/(3-catenin, which was up-regulated by TGF(31 and down-regulated by ICG-001. Immunoblotting assay also indicated that ICG-001 reduced the expressions of ubiquitin and (3-catenin in TGF(31-treated HConFs, implying that CRYAB stabilized (3-catenin by inhibiting its ubiquitination degradation. Exogenous CRYAB promoted cell viability, increased extra- cellular matrix protein levels, and up-regulated a-SMA expression of HConFs under TGF(31 stimulation. CRYAB rescued TGF(31-induced fi brotic responses that were suppressed by ICG-001. In conclusion, this study elucidates the regulatory mechanism of the Wnt/(3-catenin/CRYAB pathway in conjunctival fi brosis, offering promising therapeutic targets for mitigating bleb scarring after glaucoma fi ltration surgery. (Am J Pathol 2024, 194: 1764-1779; https://doi.org/10.1016/j.ajpath.2024.05.002)

Background: Colorectal cancer (CRC) is the third most common malignant tumor worldwide. Angiogenesis is closely related to tumor metastasis, which is the main cause of cancer death. Although several angiogenesis signatures have been proposed in some cancer types, no angiogenic signature has been developed to predict the prognosis and efficacy of antiangiogenic bevacizumab in CRC patients. Methods: We developed a novel CRC angiogenic signature by refining seven publicly available angiogenic gene sets using least absolute shrinkage and selection operator (LASSO). Immune and stromal cells within the tumor microenvironment were compared between the high- and low-risk groups in more than 1000 CRC samples classified by calculating the risk score based on the customized angiogenic signature. The correlation of this new gene set with the efficacy of bevacizumab was also compared. Results: A new prognostic-associated angiogenesis signature gene set was constructed that can divide CRC patients into two high- and low-risk groups. The high-risk angiogenic group was significantly associated with extracellular matrix organization, epithelial-mesenchymal transition (EMT), and myogenesis. In addition, the high-risk group had higher infiltration of stromal and immune cells and was more resistant to bevacizumab than the low-risk group. Conclusion: Briefly, we constructed a novel angiogenic signature that can predict the prognosis of CRC patients and the efficacy of bevacizumab in treating CRC. Our results provide new insights into the relationships among angiogenesis, metastasis, and medication for CRC.

Conjunctival fibrosis is a common postoperative complication of glaucoma filtration surgery, resulting in uncontrolled intraocular pressure and surgery failure. Therefore, there is an urgent need to understand the molecular mechanisms underlying conjunctival fibrosis and to explore novel pharmacologic anti-fibrosis therapies for glaucoma filtration surgery. Herein, the 4-dimensional data-independent acquisition (4D-DIA) quantitative proteomic results, coupled with experimental data, revealed the activation of the Wnt/β-catenin pathway in transforming growth factor (TGF)-β1-induced human conjunctival fibroblasts (HConFs). Treatment with ICG-001, a Wnt/β-catenin inhibitor, effectively inhibited cell proliferation and migration in TGFβ1-treated HConFs. ICG-001 treatment alleviated the increased generation of extracellular matrix proteins induced by TGFβ1. In addition, ICG-001 reduced the expression level of α smooth muscle actin (α-SMA) and inhibited cell contractility in TGFβ1-treated HConFs. Proteomics data further suggested that αB-crystallin (CRYAB) was a downstream target of Wnt/β-catenin, which was up-regulated by TGFβ1 and down-regulated by ICG-001. Immunoblotting assay also indicated that ICG-001 reduced the expressions of ubiquitin and β-catenin in TGFβ1-treated HConFs, implying that CRYAB stabilized β-catenin by inhibiting its ubiquitination degradation. Exogenous CRYAB promoted cell viability, increased extracellular matrix protein levels, and up-regulated α-SMA expression of HConFs under TGFβ1 stimulation. CRYAB rescued TGFβ1-induced fibrotic responses that were suppressed by ICG-001. In conclusion, this study elucidates the regulatory mechanism of the Wnt/β-catenin/CRYAB pathway in conjunctival fibrosis, offering promising therapeutic targets for mitigating bleb scarring after glaucoma filtration surgery.

AIM: To identify genetic defects in a Chinese family with congenital posterior polar cataracts and assess the pathogenicity. METHODS: A four-generation Chinese family affected with autosomal dominant congenital cataract was recruited. Nineteen individuals took part in this study including 5 affected and 14 unaffected individuals. Sanger sequencing targeted hot-spot regions of 27 congenital cataract-causing genes for variant discovery. The pathogenicity of the variant was evaluated by the guidelines of American College of Medical Genetics and InterVar software. Confocal microscopy was applied to detect the subcellular localization of fluorescence-labeled ephrin type-A receptor 2 (EPHA2). Co-immunoprecipitation assay was implemented to estimate the interaction between EphA2 and other lens membrane proteins. The mRNA and protein expression were analyzed by reverse transcription-polymerase chain reaction (qRT-PCR) and Western blotting assay, respectively. The cell migration was analyzed by wound healing assay. Zebrafish model was generated by ectopic expression of human EPHA2/p.R957P mutant to demonstrate whether the mutant could cause lens opacity in vivo. RESULTS: A novel missense and pathogenic variant c.2870G>C was identified in the sterile alpha motif (SAM) domain of EPHA2. Functional studies demonstrated the variant’s impact: reduced EPHA2 protein expression, altered subcellular localization, and disrupted interactions with other lens membrane proteins. This mutant notably enhanced human lens epithelial cell migration, and induced a central cloudy region and roughness in zebrafish lenses with ectopic expression of human EPHA2/p.R957P mutant under differential interference contrast (DIC) optics. CONCLUSION: Novel pathogenic c.2870G>C variant of EPHA2 in a Chinese congenital cataract family contributes to disease pathogenesis.

Purpose: To investigate the potential effects and mechanism of nicotinamide riboside (NR) on the oxidative stress and fibrosis model of human trabecular meshwork (HTM) cell line cells. Methods: HTM cells were pretreated with NR, followed by the induction of oxidative injury and fibrosis by hydrogen peroxide (H 2 O 2 ) and TGF- 0 2, respectively. Cell viability was tested using Hoechst staining and MTT assays, cell proliferation was assessed by EdU assay, and cell apoptosis was detected by flow cytometry and western blotting. DCFH-DA and DHE probes were used to measure the level of reactive oxygen species (ROS), and MitoTracker staining was used to measure the mitochondrial membrane potential (MMP). Fibrotic responses, including cell migration and deposition of extracellular matrix (ECM) proteins, were detected via Transwell assays, qRT-PCR, and immunoblotting. Results: NR pretreatment improved the viability, proliferation, and MMP of H 2 O 2 -treated HTM cells. Compared to cells treated solely with H 2 O 2 , HTM cells treated with both NR and H 2 O 2 , exhibited a reduced rate of apoptosis and generation of ROS. Compared with H 2 O 2 pretreatment, NR pretreatment upregulated expression of the JAK2/Stat3 pathway but inhibited mitogen-activated protein kinase (MAPK) pathway expression. Moreover, 10-ng/mL TGF- 0 2 promoted cell proliferation and migration, which were inhibited by NR pretreatment. Both qRT-PCR and immunoblotting showed that NR inhibited the expression of fibronectin in a TGF- 0 2-induced fibrosis model. Conclusions: NR has a protective effect on oxidative stress and fibrosis in HTM cells, which may be related to the JAK2/Stat3 pathway and MAPK pathway. Translational Relevance: Our research provides the ongoing data for potential therapy of NAD + precursors in glaucoma.

Retinal neurodegenerative diseases are a category of refractory blinding eye conditions closely associated with oxidative stress induced by mitochondrial dysfunction in retinal cells. SARM1, a core driver molecule leading to axonal degeneration, possesses NAD+ enzyme (NADase) activity. However, the role of the SARM1-NAD+ axis in oxidative stress-induced retinal cell death remains unclear. Here, we employed the SARM1 NADase inhibitor DSRM-3716 and established a glucose oxidase (GOx)-induced oxidative stress cell model. We found that compared to the GOx group, the DSRM-3716 pre-treated group reduced the hydrolysis of NAD+, inhibited the elevation of oxidative stress markers induced by GOx, decreased mitochondrial dysfunction, lowered the phosphorylation level of JNK, and attenuated the occurrence of pyroptosis in retinal and nerve cells, thereby providing protection for neurite growth. Further utilization of the JNK activator Anisomycin activated JNK, revealed that the JNK/c-Jun pathway down-regulated NMNAT2 expression. Consequently, it reduced cellular NAD+ synthesis, exacerbated mitochondrial dysfunction and cell pyroptosis, and reversed the protective effect of DSRM-3716 on cells. In summary, the inhibition of SARM1 NADase activity substantially mitigates oxidative damage to retinal cells and mitochondrial damage. Additionally, JNK simultaneously serves as both an upstream and downstream regulator in the SARM1-NAD+ axis, regulating retinal cell pyroptosis and neurite injury. Thus, this study provides new insights into the pathological processes of retinal cell oxidative stress and identifies potential therapeutic targets for retinal neurodegenerative diseases.

PURPOSE:To identify RHO mutations in patients with non-syndromic retinitis pigmentosa (NS-RP).METHODS:A total of 143 probands (46 family history and 97 sporadic cases) with NS-RP were recruited from Southeast China. The coding exons and adjacent intronic regions of RHO were PCR-amplified and sequenced by Sanger sequencing. The candidate variant was evaluated by the guidelines of American College of Medical Genetics and further validated through co-segregation analysis within the family.RESULTS:Five heterozygous mutations in RHO were detected in 5 out of 143 probands, where the frequency of RHO mutations in our cohort was approximately 3.5% (5/143) and 10.8% (5/46) for probands and families with NS-RP, respectively. Three known disease-causing mutations including c.C1030T (p.Q344X), c.C173G (p.T58R), and c.G266A (p.G89D) were identified in three unrelated families. The other two previously unreported mutations c.557C>A (p.S186X) and c.944delA (p.N315TfsX43) were confirmed in Family RP-087 and Family RP-139, respectively. These mutations co-segregated with available affected individuals in each family were not observed in the unaffected family members or in the 112 unrelated controls.CONCLUSIONS:This report expands the mutational spectrum of RHO gene associated with NS-RP and demonstrates the frequency of RP RHO mutations in Southeast Chinese populations.

To observe the safety and effect of phacoemulsification combined with intraocular lens (IOL) implantation in patients with low corneal endothelial cell density (CD) under the low perfusion pattern with low negative pressure.In this retrospective case series study, a total of 16 patients (17 eyes) were studied. They had all been diagnosed with low corneal endothelial (CD) and cataracts in the First Affiliated Hospital of Fujian Medical University from December 2019 to October 2021. They underwent phacoemulsification combined with IOL implantation under the low perfusion pattern with low negative pressure. The variations of corneal endothelial( CD), coefficient of variation (CV) of the cell area, central corneal thickness (CCT), visual acuity, and intraocular pressure before and after the operation were observed and assessed. Then a paired t-test, repeated measures analysis of variance, and Pearson correlation analysis were adopted for data analysis.The mean intraocular pressure of the 17 eyes was 16.88 ± 6.47 mmHg before the operation and 14.41 ± 3.10 mmHg after the operation, showing a statistically significant difference of t = 2.222, and p = 0.041. Before the operation, the mean visual acuity was 0.16 ± 0.09, and after the operation, it was 0.45 ± 0.16, displaying a statistically significant difference of t = -9.917, p < 0.001. Before and after the operation, four of the 17 eyes had no detectable CD. The mean CD of the other 13 eyes at one month after the operation (644.308 ± 106.24 cells/mm2) was lower than that before the operation (709.62 ± 119.19 cells/mm2), and the differences were statistically significant (F = 20.044, p < 0.001). However, no statistically significant differences were found in the mean CV before the operation (31.23 ± 4.21), and at one month after the operation (32.62 ± 3.80; F = 2.130, p = 0.157). Moreover, the mean CCT of 14 eyes at one month after the operation (562.72 ± 27.82 μm) was larger than that before the operation (534.79 ± 24.69 μm).The low perfusion pattern with low negative pressure is safe and effective for corneal endothelial dysfunction patients complicated with cataracts.

The outcome of neoadjuvant chemoradiotherapy (nCRT) remains highly unpredictable for individuals with locally advanced rectal cancer (LARC). We set out to characterize effective biomarkers that promote a patho-logical complete response (pCR). We quantified the abundances of 6483 high-confidence proteins in pre-nCRT biopsies of 58 LARC patients from two hospitals with pressure cycling technology (PCT)-assisted pulse data -independent acquisition (PulseDIA) mass spectrometry. Compared with non-pCR patients, pCR patients ach-ieved long-term disease-free survival (DFS) and had higher tumor immune infiltration, especially CD8+ T cell infiltration, before nCRT. FOSL2 was selected as the candidate biomarker for predicting pCR and was found to be significantly upregulated in pCR patients, which was verified in another 54 pre-nCRT biopsies of LARC patients by immunohistochemistry. FOSL2 expression was able to predict pCR by multiple reaction monitoring (MRM) with high efficiency (Area under curve (AUC) = 0.939, specificity = 1.000, sensitivity = 0.850), and high FOSL2 expression was associated with long-term DFS (p = 0.044). When treated with simulated nCRT, FOSL2 suffi-ciency resulted in more significant inhibition of cell proliferation, and more significant promotion of cell cycle arrest and cell apoptosis. Moreover, CXCL10 secretion with abnormal cytosolic dsDNA accumulation was found in FOSL2-wildtype (FOSL2-WT) tumor cells over nCRT, which might elevate CD8+ T-cell infiltration and CD8+ T -cell-mediated cytotoxicity to promote nCRT-induced antitumor immunity. Our study revealed proteomic profiles in LARC patients before nCRT and highlighted immune activation in the tumors of patients who achieved pCR. We identified FOSL2 as a promising biomarker to predict pCR and promote long-term DFS by contributing to CD8+ T-cell infiltration.

Introduction: Hermansky-Pudlak syndrome (HPS) is a rare autosomal-recessive disease characterized by ocular albinism (OA) or oculocutaneous albinism (OCA), platelet dysfunction, and other symptoms. This study aimed to analyze the molecular defect in two Chinese families with suspected OA, as well as to investigate the profile of HPS6 variants and their genotype-phenotype correlations. Methods: Seven members from two families were recruited and underwent clinical ophthalmologic examinations. The genomic DNA was extracted from peripheral blood leukocytes. Whole-exome sequencing was performed on the proband of family JX. The single coding exon of HPS6 was directly Sanger sequenced based on PCR amplification in all available family members. An additional 46 probands from families or sporadic cases with the pathogenic variants of HPS6 reported in the literature were reviewed. Results: We identified two different compound heterozygous truncating variants of HPS6 in probands with suspected OA from two independent families. The proband of family JX had c.1674dup and c.503-504del variants, and the other proband from family CZ had a nonsense variant of c.1114C>T and a frameshift variant of c.1556del. Among them, c.1674dup and c.1556del variants in HPS6 have not been reported previously. Therefore, our patients were diagnosed as HPS6 disease by molecular diagnostics. In the retrospective cohort of HPS6 patients, we delineated the profile of HPS6 variants and revealed a significant overlap between CpG islands and the variants of HPS6, suggesting a potential link between DNA methylation and HPS6 variants. We also observed a spatial aggregation of the variants in 3D structure of HPS6 protein, implying the possible functional significance of these structural regions. In addition, we did not find any significant genotype-phenotype correlation of HPS6, and neither did we observe a correlation between the truncation length of the HPS6 protein and the phenotype of HPS6 disease. Conclusion: Our research expands the spectrum of HPS6 variants, providing a comprehensive delineation of their profile and systematically investigating genotype-phenotype correlations in HPS6. These findings could offer potentially valuable clues for investigating the molecular mechanism underlying HPS6 pathogenesis, as well as aiding the clinical diagnosis of HPS6 patients and improving disease prognosis.

MYOC is a common pathogenic gene for primary open-angle glaucoma and encodes the protein named myocilin. Multiple MYOC variations have been found, with different clinical significance. However, the pathogenesis of glaucoma induced by MYOC mutations has not been fully clarified. Here, we analyze the molecular and cellular biological differences caused by multiple variant myocilins, including protein secretion characteristics, structural changes, subcellular localization, cellular autophagic activity and oxidative stress. Denaturing and nondenaturing electrophoresis showed myocilin to be a secreted protein with the tendency to self-oligomerize. The full-length myocilin and its C-terminal cleavage fragment are secreted. Secretion analysis of 23 variant myocilins indicated that secretion defects are closely related to the pathogenicity of MYOC variants. Structural analysis showed that the alteration of steric clash is associated with the secretion characteristics and pathogenicity of myocilin variants. Immunocytochemistry results demonstrated that mutated myocilins are retained in the endoplasmic reticulum and disrupt autophagy. MTT assay, MitoTracker staining, and DCFH-DA staining showed increased oxidative injury in cells expressing MYOC mutants. Taken together, MYOC mutations are able to induce cell dysfunction via secretion defects and intracellular accumulation resulting from steric clash alterations.