Colletotrichum is a relatively uncommon cause of human infection, and previous findings on Colletotrichum keratitis have been scarce. This research describes the clinical characteristics and treatment outcomes of 9 cases of Colletotrichum keratitis discovered in our hospital using next-generation sequencing (NGS). Among 78 patients with NGS-proven fungal keratitis, 9 (11.5%) were infected with Colletotrichum species. NGS revealed that 3 patients were infected with C. truncatum, 3 with C. gloeosporioides, and 3 with C. fructicola. Seven patients had a history of corneal plant trauma, one patient was infected by mosquitoes, and one patient had an unknown origin. Seven patients underwent penetrating keratoplasty, and two underwent lamellar keratoplasty. Eight patients healed after keratoplasty, but one required evisceration due to recurrence of fungal infection. NGS allows for more precise diagnosis and enhances awareness of Colletotrichum keratitis, which is not as rare as previously reported.
Pluripotent stem cells, having long been considered the fountain of youth, have caught the attention of many researchers from diverse backgrounds due to their capacity for unlimited self-renewal and potential to differentiate into all cell types. Over the past 15 years, the advanced development of induced pluripotent stem cells (iPSCs) has displayed an unparalleled potential for regenerative medicine, cell-based therapies, modeling human diseases in culture, and drug discovery. The transcription factor quartet (Oct4, Sox2, Klf4, and c-Myc) reprograms highly differentiated somatic cells back to a pluripotent state recapitulated embryonic stem cells (ESCs) in different aspects, including gene expression profile, epigenetic signature, and functional pluripotency. With the prior fruitful studies in SCNT and cell fusion experiments, iPSC finds its place and implicates that the differentiated somatic epigenome retains plasticity for re-gaining the pluripotency and further stretchability to reach a totipotency-like state. These achievements have revolutionized the concept and created a new avenue in biomedical sciences for clinical applications. With the advent of 15 years' progress-making after iPSC discovery, this review is focused on how the current concept is established by revisiting those essential landmark studies and summarizing its current biomedical applications status to facilitate the new era entry of regenerative therapy.
Abstract In this article, human limbal niche cells (LNC) or bone marrow derived mesenchymal stem cells (BMMSC) were used to prevent limbal stem cell deficiency (LSCD) in an alkali burn rabbit model and their results were compared. The epithelial cell defect area, corneal neovascularization, and the print cell cytometry were quantified to grade the severity of LSCD. Three months after the alkali burn, a partial LSCD was observed in the control group (no treatment) indicated by chronic corneal epithelial defects, positive corneal fluorescein staining, neovascularization and goblet cell migration. In contrast, the severity of LSCD in both the LNC and BMMSC transplantation groups was dramatically reduced as shown by smaller epithelial cell defects, decreased fluorescein sodium staining, decreased neovascularization and decreased goblet cell density. Interestingly, the LNC group was shown to more effectively prevent LSCD than the BMMSC group. Further analysis indicated subconjunctivally transplanted LNCs were more powerful than BMMSCs to prevent LSCD, at least partially, due to increased activation of SCF-c-Kit signal. We conclude that LNCs are a more powerful resource than BMMSCs to prevent LSCD in an alkali burn rabbit model, at least partially due to increased activation of SCF signaling.
The efficacy and safety of 3% diquafosol sodium eye drops in Chinese patients with dry eye in the real-world setting remains unclear.3099 patients with dry eye symptoms were screened according to Asia Dry Eye Society latest recommendation. Among them, 3000 patients were enrolled for a phase IV study. We followed up with multiple clinical characteristics including corneal fluorescein staining, tear break up time, Schirmer's tests, visual acuity, intraocular pressure, and others. The follow ups were performed at baseline, 2 weeks and 4 weeks after treatment.Based on the results of corneal fluorescein staining and tear break up time, all age and gender subgroups exhibited obvious alleviation of the symptoms among the patients with dry eye, and the data in elderly group showed the most significant alleviation. All the adverse drug reactions (ADRs, 6.17%) were recorded, among which 6% local ocular ADRs were included. Meanwhile, mild ADRs (91.8%) accounted for the most. Most of the ADRs (89.75%) got a quick and full recovery, with an average time at 15.6 days. 1.37% of patients dropped out of the study due to ADRs.The use of 3% diquafosol sodium eye drop is effective and safe in the treatment of dry eye, with a low incidence of ADRs showing mild symptoms. This trial was registered at Chinese Clinical Trial Registry ID: ChiCTR1900021999 (Registration Date: 19/03/2019).
Objective To explore an improved method to culture corneal endothelial cells.Methods Through surgical microscope,Descemet membrane and endothelial cell layer were isolated and overlaid on cover glasses,then dissected and seeded in culture containers with endothelial cell layer surface upwards.Results The cells grew well without adding any mitogen in the tissue culture medium.The corneal endothelial cells grew and were confluent within two weeks,the corneal endothelial cells could successfully obtain.Conclusion Enzyme is not required in this method;moreover,the method has many advantages,such as unfold tissue pieces,firm attachment,easy migration,and fast proliferation.With this technique,sufficient quantities of normal live cells become available for research of corneal endothelium.
Background: Fungal keratitis (FK) is a leading cause of corneal blindness in developing countries due to poor clinical recognition and laboratory identification. Here, we aimed to identify the distinct clinical signature of FK and develop a diagnostic model to differentiate FK from other types of infectious keratitis.Methods: We reviewed the electronic health records (EHRs) of all patients with suspected infectious keratitis in Beijing Tongren Hospital from January 2011 to December 2021. Twelve clinical signs were assessed by Lasso regression analysis and collinear variables were excluded. Three models based on binary logistic regression, random forest classification, and decision tree classification were trained for FK diagnosis and employed for internal validation. Independent external validation of the models was performed in a cohort of 420 patients across China to evaluate the accuracy, specificity and sensitivity in real world.Findings: Three diagnostic models of FK based on binary logistic regression, random forest classification, and decision tree classification were established and internal validation were achieved with the mean AUC of 0.916, 0.920, and 0.859, respectively. The models were well-calibrated by external validation using a prospective cohort including 210 FK and 210 non-FK patients from seven eye centers across China. The diagnostic model with the binary logistic regression algorithm classified the external validation dataset with a sensitivity of 0.907 (0.774, 1.000), specificity 0.899 (0.750, 1.000), accuracy 0.905 (0.805, 1.000), and AUC 0.903 (0.808, 0.998).Interpretation: Our model enables rapid identification of FK, which will help ophthalmologists to establish a preliminary diagnosis and to improve the diagnostic accuracy in clinic.Funding Information: This study was supported by The Open Research Fund from the National Key Research and Development Program of China (2021YFC2301000); the Open Research Fund from Beijing Advanced Innovation Center for Big Data-Based Precision Medicine, Beijing Tongren Hospital, Beihang University & Capital Medical University (BHTR-KFJJ-202001).Declaration of Interests: The authors declare that they have no competing interests.Ethics Approval Statement: Ethical approval was obtained from the Medical Ethics Committee of Beijing Tongren Hospital (TRECKY2021-024) and all other participating hospitals, including, Peking Union Medical College Hospital (No. zs-3516), the First Affiliated Hospital of Chongqing Medical University (No. 2014- 76), the Second Xiangya Hospital of Central South University (No. LYF2021028), Eye Hospital of Wenzhou Medical College (No. 2020-191-K-174), and Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology (No. 2020-120), Peking University People's Hospital (No. 2017PHB167-01), Eye & ENT Hospital of Fudan University (No. 2021103). The study protocol fully adhered to the Declaration of Helsinki and the ARVO statement on human subjects. All subjects finished the written informed consent documents. If participants were under 18 years old, written informed consent should be got from their parents.
Objective To study the influence of water soluble azone on proliferation and ultrastructure of human trabecular mashwork cells (HTMC) .Methods Human trabecular mashwork cells were cultured and passaged in vitro. The cells were treated with different concentra tions of azone, MTT assay, transmission electron microscope and acridine orange fluorescent staining were used to observe the influence of azone on the proliferation and ultrastructure of HTMC cells. Results 10-5to 10- 8 g/L of azone treaded for 24 h had no significant influence on the proliferation and vitality of HTMC cells, 10-5 g/L of water soluable azone had no significant influence on the proliferation of HTMC cells. 10 -6 to 10-8 g/L of azone treaded for 12 h had no significant influence on ultrastructure of HTMC cells. Acridine orange fluorescent staining showed that HTMC cells treated with 10-3 g/L or 10-4 g/L of azone for 6 h lost its vitality in some degree. C ondusion 10-6 to 10-8 g/L of water soluable azone had no significant influence on the proliferation, vitality and ultrastructure of HTMC cells. Azone may be used widely in the eye in the future.
Key words:
azone/administration dosage; human trabecular mashwork cells/ultrastructure; proliferation; vitality
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