Analysis of HLA Profiles in 390 Uveitis Cases: Insights into Clinical Presentations.

Objective: This study aimed to explain the associations between different types of uveitis and human leukocyte antigen (HLA)-B27, HLA-DR4, and HLA-DRw53. Methods: A retrospective analysis of 390 uveitis cases was conducted among inpatients and outpatients diagnosed at Weifang Eye Hospital from 2013 to 2016. All 390 patients underwent HLA-B27 examination, and an additional 40 patients underwent examination for HLA-DR4 and HLA-DRw53. Gender, age, corrected visual acuity (CVA), and recurrence frequency were statistically analyzed based on the onset site and etiology classification. Results: Among the 390 enrolled patients, 206 were male, and 183 were female, with ages ranging from 6 to 87 years (mean: 44.2). The disease onset was classified into anterior uveitis (AU), panuveitis (panU), posterior uveitis (PU), and intermediate uveitis in 180, 112, 88, and 10 cases, respectively. HLA-B27 was positive in 94 cases (53 males and 41 females), yielding a positive rate of 24.1%. In AU patients, 80 (44.4%) tested positive for HLA-B27, while 8 (7.1%) panU patients and 6 PU patients (6.8%) were HLA-B27 positive; none of the intermediate uveitis (IU) patients exhibited HLA-B27 positivity. HLA-B27, HLA-DR4, and HLA-DRw53 examinations were performed on 40 patients with binocular uveitis, resulting in 2 HLA-B27 positive cases, 15 HLA-DR4 positive cases, and 20 HLA-DRw53 positive cases, with positive rates of 5%, 37.5%, and 50%, respectively. Among 25 Vogt Koyanagi-Harada (VKH) cases, 1 tested positive for HLA-B27, 22 were positive for HLA-DR4, and 24 were positive for HLA-DRw53, with positive rates of 4%, 88%, and 96%, respectively. No positive HLA-B27, HLA-DR4, or HLA-DRw53 cases were found among the 10 cases of Behcet's disease (BD). Conclusions: Human leukocyte antigens (HLAs) play a significant role in the mechanism of uveitis. HLA-B27 exhibits high diagnostic value in acute AU, while HLA-DR4 and HLA-DRw53 are crucial for diagnosing binocular uveitis, particularly Vogt Koyanagi-Harada (VKH) syndrome. Further investigation is warranted to explore the relationship between HLA-DR4, HLA-DRw53, and uveitis.

KLF3 Transcription Activates WNT1 and Promotes the Growth and Metastasis of Gastric Cancer via Activation of the WNT/ß-Catenin Signaling Pathway.

The transcription factor Krüppel-like factor (KLF) 3 is one of the members of the KLF family, which plays an important role in tumor progression. Nevertheless, the role of KLF3 in the growth and metastasis of gastric cancer (GC) still needs to be elucidated. Bioinformatics analysis showed that KLF3 was overexpressed in patients with GC, and the high expression of KLF3 was correlated with poor survival. KLF3 was also overexpressed in GC clinical samples and cell lines. In vitro functional role of KLF3 in GC cells was explored by a gain-of-function and loss-of-function assay. Overexpressed KLF3 promoted the cell proliferation, migration, invasion, and epithelial-mesenchymal transition of GC cells, whereas suppressed KLF3 inhibited these biological behaviors. The clinical samples and bioinformatics analysis showed that WNT1 was also highly expressed in GC tumor tissues and positively correlated with KLF3 expression. The luciferase reporter assay and chromatin immunoprecipitation result confirmed that KLF3 could directly bind to the WNT1 promoter to increase the transcriptional activity of WNT1, thus regulating its expression. Overexpressed KLF3 enhanced the protein expression level of p-GSK3ß(Ser9) and ß-catenin, the key elements in the WNT/ß-catenin signaling pathway. Repression of KLF3 decreased the level of p-GSK3ß(Ser9) and ß-catenin. Immunofluorescence images showed that KLF3 promoted nuclear ß-catenin accumulation. Inhibition of WNT1 attenuated the proliferation, migration, and invasiveness of KLF3-overexpressing GC cells. Moreover, the xenograft mouse model confirmed that KLF3 promotes GC tumor growth and metastasis in vivo. Our results demonstrated that KLF3 activates the WNT/ß-catenin signaling pathway via WNT1 to promote GC tumor growth and metastasis, indicating that repression of KLF3 may act as a potential therapeutic target for patients with GC.

The Molecular Mechanism of Long Non-Coding RNA (LncRNA) Regulation of Notch Signaling in Glucose-Induced Apoptosis of Human Retinal Vascular Endothelial Cell.

Diabetes, a global health concern, affects the health of more than 500 million adults. The absence of Notch protein can cause an imbalance in the retinal vascular environment and cause retinal vascular disease. Long noncoding RNA (lncRNA) is known to be involved in the regulation of many signaling pathways. We hope to understand the specific mechanism of apoptosis in retinal vascular endothelial cells (RVECs) by exploring the regulatory effect of lncRNA on the Notch pathway. In this study, we found that RVECs treated with glucose showed increased levels of Notch transcript and protein expression. The lentiviral interference with Notch RNAi reversed this response. When Notch activity decreased, oxidative stress also decreased, accompanied by increased levels of Caspase-9 and Caspase-3 and an increased rate of apoptosis. Therefore, we believe that Notch is involved in the development of diabetic retinopathy and loss of expression promotes apoptosis of human RVECs. By inhibiting the Notch pathway, lncRNA promotes apoptosis of human RVECs in a high-glucose environment.

A meta-analysis of prognostic biomarkers in neonatal retinal hemorrhage.

PURPOSE: Neonatal retinal hemorrhage (RH) is a frequently occurring neonatal fundus condition and a very common ocular abnormality in neonates. Some of the key factors that influence the rate of RH are the mode of delivery, examination techniques, and time of examination after birth. The prognostic markers of severe RH are poorly known, making it difficult for an efficient diagnosis, prognosis, and treatment. Hence, to better understand the mechanism of disease, its study at the molecular level is required. Prognostic biomarkers are an essential tool for understanding the pathogenesis of the disease. In this paper, we present a meta-analysis of biomarkers to understand disease pathogenesis and support better diagnosis, prognosis, and treatment of neonatal RH. METHODS: The meta-analysis was carried out by following the recommendation of PRISMA. The relevant articles were crawled using a systematic keyword using MeSH terms from the MEDLINE, PubMed, and Scopus databases, which were subjected to manual screening for reported biomarkers by two independent reviewers. The obtained biomarkers were further analyzed for gene-disease association and functional enrichment analysis. RESULTS: Our meta-analysis suggests that genes ABCC6, Beta-APP, COL2A1, COL4A1, DNM2, ENPP1, IKBKG, ITGB2, IL-6, SELE, TREX1, and VEGFA are potential prognostic biomarkers associated with the neonatal RH. The gene-disease association and functional enrichment analysis suggest that few genes are associated with disease class "Vision"; however, some genes in the list are associated with the disease class "Pharmacogenomic," "Immune," "Renal." CONCLUSION: The identified prognostic gene biomarkers may help to understand disease pathogenesis and provide a better diagnosis, prognosis, and treatment of neonatal RH.

A review of sample thickness effects on high-resolution transmission electron microscopy imaging.

High-resolution transmission electron microscopy (HRTEM) is an important approach to analyzing material structures. However, in reality, preparing a sufficiently thin sample for use in HRTEM, based on which images could be interpreted by weak phase object approximation theory, is difficult. During the imaging process, the thickness of the sample has two primary effects-a dynamical effect and a non-linear effect. Both are reviewed in this paper. Considering only the dynamical effect, the Bloch wave method and multislice theory have been proposed to understand the relationship between sample thickness and imaging. These methods exhibit high accuracy but high complexity as well. Sacrificing accuracy, pseudo-weak phase object approximation (PWPOA) theory can provide clues to the relationship in reciprocal space with greater simplicity. Meanwhile, in real space, channeling theory describes the dynamical effect with sufficient accuracy, and with the 1s state approximation, i.e., for a certain range of thicknesses, it provides a physical image and simplified expression with which to describe the relationship between the exit wave and sample thickness. As for the non-linear effect, a method of separating linear and non-linear information using a combination of transmission cross-coefficient theory and PWPOA theory was recently proposed. The variation of non-linear and linear imaging with sample thickness has also been discussed. A deep understanding has been acquired regarding the effects of the sample thickness, but a complete understanding of the HRTEM imaging process for thick samples has remained elusive. This understanding is crucial to the retrieval of structure from HRTEM images.

Leber's Hereditary Optic Neuropathy-Specific Heteroplasmic Mutation m.14495A>G Found in a Chinese Family.

PURPOSE: Leber's hereditary optic neuropathy (LHON) is a mitochondrial DNA (mtDNA)-associated, maternally inherited eye disease. Mutation heteroplasmy level is one of the leading causes to trigger LHON manifestation. In this study, we aimed to identify the causative mutation in a large Han Chinese family with LHON and explore the underlying pathogenic mechanism in this LHON family. METHODS: The whole-mtDNA sequence was amplified by long-range PCR. Mutations were subsequently identified by next-generation sequencing (NGS) and validated by Sanger sequencing. The heteroplasmy rates of those family members were determined by digital PCR (dPCR). Mitochondrial haplogroups were assigned based on mtDNA tree build 17. RESULTS: The m.14495A>G mutation was identified as causative due to its higher heteroplasmy level (>50%) in patients than in their unaffected relatives. All mutation carriers belong to M7b1a1 and are assigned to Asian mtDNA lineage. Interestingly, our result revealed that high mtDNA copy number in carrier might prevent LHON manifestation. CONCLUSIONS: This is the first report of m.14495A>G mutation in Asian individuals with LHON. Our study shows that dPCR technology can provide more reliable results in mutation heteroplasmy assay and determination of the cellular mtDNA content, making it a potentially promising tool for clinical precise diagnosis of LHON. Furthermore, our results also add evidence to the opinion that higher mtDNA content may protect mutation carriers from LHON. TRANSLATIONAL RELEVANCE: dPCR can be used for the assessment of LHON disease, and a new genetic-based diagnostic strategy has been proposed for LHON patients with the m.14495A>G mutation.

ABT737 enhances ovarian cancer cells sensitivity to cisplatin through regulation of mitochondrial fission via Sirt3 activation.

AIMS: The poor prognosis of ovarian cancer is mainly caused by chemotherapy resistance. Studies show that the Bcl-2 inhibitor ABT737 can significantly improve the effect of cisplatin and induce mitochondrial pathway apoptosis. However, the mechanism of ABT737 increases sensitivity to cisplatin by regulating mitochondrial function remains unclear in ovarian cancer cells. Sirt3, as a histone deacetylase, is involved in the regulation of mitochondrial function in cancers. In this study, we intend to explore the mechanistic link between Sirt3 and mitochondrial dysfunction induced by ABT737 and cisplatin in ovarian cancer cells. MAIN METHODS: Apoptosis was examined by flow cytometry following Annexin V and PI staining. Sirt3 activity was assessed using Sirt3 deacetylase fluorometric assay. The mitochondrial membrane potential was examined by flow cytometry following JC-1 staining. Overexpression and knock-down of Sirt3 were confirmed by western blot analysis. Mitochondrial fission/fusion dynamics were detected by immunofluorescence staining or western blot analysis. KEY FINDINGS: Cisplatin accompanied with ABT737 promoted apoptosis and decreased mitochondrial membrane potential. ABT737 enhanced the sensitivity of ovarian cancer cells to cisplatin, which was partly achieved by activating Sirt3 to regulate the mitochondrial fission process. SIGNIFICANCE: This study identified the activation of Sirt3 played an important role in increasing sensitivity of ovarian cancer cells to cisplatin induced by ABT737. Furthermore, Sirt3 might represent a potential therapeutic target for ovarian cancer.

Applicability of non-linear imaging in high-resolution transmission electron microscopy.

According to transmission cross-coefficient theory, the information limit of non-linear imaging in high-resolution transmission electron microscopy is, under certain conditions, far beyond that of linear imaging, which suggests the possibility of using high-frequency information for structural determination. In this article, we studied the information beyond the linear information limit by means of multislice method simulation, with AlN as an example, and more structural information was obtained by using part of the high-frequency information.

MiR-199a/b-3p suppresses migration and invasion of breast cancer cells by downregulating PAK4/MEK/ERK signaling pathway.

MicroRNA-199a/b-3p is downregulated in several types of aggressive cancer, and its decrement significantly correlates with poor survival. Here, we aim to investigate the biological function of miR-199a/b-3p and its regulation of target genes in breast cancer cells with highly metastatic potential. In addition, we found that miR-199a/b-3p expression was much lower in MDA-MB-231, CAL120, and HCC1395 breast cancer cells with highly metastatic potential. Functional assays showed that restored miR-199a/b-3p expression inhibited MDA-MB-231 cell growth, cell-cycle progression, migration, and invasion. In addition, we experimentally demonstrated that PAK4 was the direct target of miR-199a/b-3p, hypo-expression of PAK4 suppressed proliferation, migration and invasion of MDA-MB-231 cells, and overexpression of PAK4 significantly rescued the inhibitory effect of miR-199a/b-3p on MDA-MB-231 cell growth, migration, and invasion. Further, we also observed that miR-199a/b-3p could inactivate the PAK4/MEK/ERK signaling pathway. Thus, miR-199a/b-3p functions as a tumor suppressor and has an important role in breast cancer metastasis through PAK4/MEK/ERK signaling pathway.

Identification of a necessary element for Toxoplasma gondii SAG1 gene expression.

SAG1 codes for the stage-specific major surface antigen P30 of Toxoplasma gondii (T. gondii) tachyzoites. Six tandemly repeated, conserved 27 bp cassettes in the region from -231 to -70 bp were previously confirmed to be essential for high-level expression of SAG1 and serve as a positioning element directing the initiation of transcription. We demonstrate here that an element located between +19 and +28 bp is necessary for SAG1 gene expression by using deletion mutagenesis analysis and electrophoresis mobility shift assay (EMSA). This will provide an insight into the regulatory mechanisms of SAG1 gene expression.