BACKGROUND:Surface marker molecules and differential ability of human adipose tissues-derived mesenchymal stem cells can change with the prolonged culture time.OBJECTIVE:To analyze the morphological characteristics and colony forming capacity of early cultured rabbit adipose tissues-derived mesenchymal stem cells in vitro.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Institute of Ophthalmology,and Central Laboratory,Eye and ENT Hospital,Fudan University from January to March 2008.MATERIALS:Nine female New Zealand rabbits aged three months were obtained from the Shanghai Yingenyang Rabbit Room.METHODS:Adipose tissue was harvested from the nape fat pad of the New Zealand rabbits following anesthesia.Primary cultured cells were established using type I collagenase and cell cultures were maintained with DMEM containing 0.1 volume fraction of fetal bovine serum.Cells were passaged when 80% was confluent.The second,third and fourth passages of cells were utilized for the study.MAIN OUTCOME MEASURES:The morphology of adipose tissues-derived mesenchymal stem cells was observed using inverted microscope.Cell phenotype was detected utilizing flow cytometry.The cloning capacity was determined.RESULTS:Adipose tissues-derived mesenchymal stem cells were fibroblast-like and proliferated rapidly in vitro at the second,third and fourth passage.Flow cytometry showed that adipose tissues-derived mesenchymal stem cells expressed CD29 and PCNA.The colony-forming rates were respectively(8.0?0.6)%,(6.7?0.4)% and(4.6?0.5)%(F=12.18,P

BACKGROUND: The application of adipose derived stromal cells to tissue engineering has been more and more popular around the world. Compatibility of scaffold material is the key point for its further research.OBJECTIVE: To determine the growth rules of rabbit adipose-derived stromal cells with polylactic-co-glycolic acid (PLGA).DESIGN, TIME AND SETTING: An in vitro study was performed at the Institute of Ophthalmology, Otolaryngology Hospital,Fudan University and Shanghai Tissue Engineering Center from September 2007 to March 2009.MATERIALS: Six female New Zealand rabbits aged six months were used for extraction of adipose-derived stromal cells. PLGA was provided by Sigma, USA.METHODS: Adipose tissue was harvested from the nape fat pad of the rabbits following anesthesia. Primary cultured cells were established using type I collagenase and cell cultures were maintained with DMEM containing 10% volume fraction of fetal bovine serum. Cells were passaged when 80% was confluent. The fourth passages of cells were utilized for the study. PLGA consisted of polylactic acid and polyglycolic acid as the ratio of 7:3, and the relative molecular mass was 104900.MAIN OUTCOME MEASURES: Initially, the adherent rate of cells to scaffold was detected. After one week co-culture, the scaffold bearing adipose-derived stromal cells labeled with Dio agent were investigated by fluorescence inverse microscope,scanning electron microscopy and laser scanning microscope.RESULTS: The best adherent rate of adipose-derived stromal cells with PLGA reached 99%. After one-week-incubation in vitro,cells of fiber-shaped or ovule-shaped proliferated well and exhibited stratified growth on the surface of PLGA scaffold. In addition,it also secreted visible extracellular matrices, which could be examined by scanning electron microscopic examination.Meanwhile, the adipose-derived stromal cells grew well and distributed equably inside the PLGA in terms of the investigation with laser scanning microscope.CONCLUSION: The compatibility of adipose-derived stromal cells to PLGA in vitro was well.

Standardization of residency training aims to improve the overall quality of residents and bring up high-quality medical students.Research capabilities and innovative spirit of young physicians play important roles in the sustainable development of hospitals.This article describes the experience in strengthening research training in the United States residency program,provides recommendations to resolve the problems of our residency standardized training and new ideas for the domestic medical institutions to strengthen the young physicians' sense of innovation and scientific research ability.

Endothelium keratoplasty (EK),due to its short operation time,fast postoperative healing and better postoperative visual prognosis,has gradually replaced penetrating keratoplasty in recent years as the preferred surgery for treating corneal endothelial decompensation.A recent study has reported the clinical results of the successful treatment of bullous keratopathy with human corneal endothelial cells cultured in vitro and anterior chamber injection,which opened a new era of EK.This method simplifies the operation procedure,avoids the complications related to the dislocation of corneal endothelial graft,and significantly improves the utilization rate of donor corneal materials.Nevertheless,long-term prognosis and outcome of patients undergoing this treatment remain to be clarified before introducing this approach into wider clinical practice.

Microfluidic chip or lab-on-a-chip is a multidisciplinary cross-technology. Among them, organ-on-a-chip technology enables precise regulation of cells and microenvironment at micron level.This tecnology is expected to simulate in vivo human physiology and overcome the shortcomings of traditional animal models and cell culture techniques.In ophthalmology, organ-on-a-chip models are primarily focused on creating biomimetic models of the cornea, retina and posterior chamber to study diseases such as dry eye, glaucoma, age-related macular degeneration and diabetic retinopathy.In addition, continuous monitoring and real-time diagnosis of tear and intraocular fluid biomarkers using microfluidic chips have become a current research hot topic.The microfluidic chips also have a wide range of applications in drug analysis, drug development, and drug screening.This article reviews the recent progress and shortcomings of microfluidic chip in in vitro model construction, point-of-care testing and drug development, and discusses its future development in ophthalmology.

Dry eye is becoming more common worldwide.Its pathophysiology is complicated, and its condition is chronic.Treatment options of dry eye are ineffective.As a multifactorial ocular surface disease, tear film instability, tear hyperosmolarity, ocular surface inflammation, and neurosensory abnormalities resulted from various causes are main natural pathological processes of dry eye.This multifactorial process of the disease leads to poor efficacy of single anti-inflammatory therapy.Oxidative stress is closely related to the occurrence of dry eye.During the decrease of tear film stability, reactive oxygen species produced by oxidative stress system damage the myelin sheath of ocular nerve and the lipid layer of tear film, inducing or aggravating the ocular inflammatory response.Targeting the main causes of dry eye's pathogenesis, stopping the vicious cycle of inflammatory responses in each link, and relieving patients' conditions are the main goals of antioxidant therapy.The development of anti-inflammatory and antioxidant medications is currently the main focus of international research on novel anti-dry eye medications.Some progresses have been made in the area of targeting oxidative stress biomarkers, mitochondrial targeting medications, mucin secretion, antioxidant enzymes like glycoprotein selenium and lactoferritin, as well as multifunctional nanoagents, and the antioxidant eye drops using nanomaterials have more advantages.Antioxidant treatment may be one of the potential future avenues of dry eye clinical research.Ophthalmologists and researchers should be fully aware of, pay close attention to and actively participate in investigations that are relevant to dry eye antioxidant therapy and the development of new medications.

Ocular surface inflammatory disorder (OSID) is a chronic ocular disease caused by systemic disorders or involving the local immune system.OSID induces persistent inflammatory reaction in the ocular adnexal connective tissues which in turn give rise to tear hypertonicity and ocular surface epithelial damage, leading to dry eye formation or progression.Common immune-related ocular surface diseases include vernal keratoconjunctivitis, Sj?gren syndrome, graft versus host disease, dry eye and immune-related corneal disease, all of which can significantly impact the visual function and quality of life of patients.Current treatments including the use of artificial tears and glucocorticoid eye drops are not always effective and have the risk of adverse events.Cyclosporine A (CsA) is a commonly utilized immunosuppressant that has a strong immunomodulatory effect, but its clinical application is somewhat limited due to the low permeability of its current ophthalmic dosage form.The development of CsA ophthalmic agents has changed the treatment strategy for OSID.The development of 0.1% CsA cationic emulsion has significantly improved the efficacy and safety of topical CsA treatment, which is worth the attention.In order to rationally apply 0.1% CsA cationic emulsion to OSID, ophthalmologists should fully understand the immune-related pathogenesis of each OSID and comprehend the curative effect, indication, application methods and adverse events of topical CsA treatment.

Objective:To evaluate the efficacy and safety of a China original liquid pulsation system for the treatment of meibomian gland dysfunction (MGD).Methods:A non-randomized controlled clinical trial was conducted.Twenty-two patients (44 eyes) diagnosed with MGD in Eye and ENT Hospital of Fudan University from February to August 2022 were enrolled.The patients were assigned into two groups according to their willingness.Of the 22 patients (44 eyes), 10 patients (20 eyes) in single liquid pulsation system group were treated with single liquid pulsation system for 12 minutes, and 12 patients (24 eyes) in intense pulsed light (IPL) group were treated with a course (4 times) of IPL, warm compresses and meibomian gland massage at three-week intervals.There was no difference in age and other baseline clinical indexes between the two groups (all at P<0.05). The meibum grading, quality grading of tear film lipid layer, Symptom Assessment Questionnaire in Dry Eye (SANDE) questionnaire score, first and average tear breakup time (BUT), corneal fluorescein sodium staining (CFS) score, tear meniscus height (TMH), and the area of meibomian gland loss were determined at baseline, 1 and 3 months after treatment.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Eye and ENT Hospital of Fudan University (No.2021069). Written informed consent was obtained from each patient before any medical examination. Results:Statistically significant group effects and time effects were found in the quality of tear film lipid layer ( Hgroup=4.39, P=0.036, Htime=6.30, P=0.043) and average BUT ( Fgroup=4.41, P=0.038; Ftime=4.08, P=0.049) in the two groups.The meibum grading, first BUT and TMH 1 and 3 months after treatment were significantly better than before treatment in single liquid pulsation system group (all at P<0.05). Compared with before treatment, there was no significant improvement in the meibum grading, distribution of tear film lipid, first BUT and TMH at 1 and 3 months after treatment in IPL group (all at P>0.05). In both groups, the SANDE and CFS scores 1 and 3 months after treatment were better than those before treatment, showing statistically significance (all at P<0.05). In terms of safety, neither instrument-related adverse events nor extra complaints of discomfort were reported in the single liquid pulsation system group.In both groups, the number of patients with positive CFS staining significantly decreased, and no new cases with positive CFS appeared after treatment. Conclusions:This China original liquid pulsation system is a safe and effective physical therapy in improving tear film dysfunction and ocular surface symptoms of MGD patients within 3 months after treatment.

Objective:To design a replaceable corneal scaler and explore its effect on improving the positive detection rate of corneal bacteria and the cure rate of patients with infectious keratitis.Methods:Totally 100 patients with infectious keratitis treated in the Eye&Ent Hospital of Fudan University in Shanghai from June 2020 to October 2021 were conveniently selected as the application object, 50 patients from June 2020 to January 2021 were selected as the control group, and 50 patients from February 2021 to October 2021 were selected as the experimental group. The control group used the traditional corneal scalpel for corneal scraping, and the experimental group used the replaceable corneal scalpel for corneal scraping. The positive detection rate of bacteria, the cure rate of patients as of the date of data collection, the comfort status of patients and the operation difficulty score of medical staff were compared between the two groups.Results:The 28 cases (56%) were positive in the experimental group, and 30 cases (60%) were cured as of the date of data collection; 16 cases (32%) were positive in the control group and 18 cases (36%) were cured as of the date of data collection. The difference between the two groups was statistically significant ( χ2=-2.47, -2.20, all P<0.05). The score of comfort in the experimental group was (83.92 ± 6.40) and that in the control group was (54.04 ± 10.69). The difference between the two groups was statistically significant ( t=17.14, P<0.01). The operation difficulty score of medical staff in the experimental group was (1.46 ± 0.54) and that of medical staff in the control group was (3.24 ± 0.72). The difference between the two groups was statistically significant ( t=-13.50, P<0.01). Conclusions:The use of replaceable corneal scalpel can not only improve the positive detection rate of patients, improve the treatment effect of patients, improve the comfort and satisfaction of patients, but also improve the work efficiency of medical staff and reduce the operational risk of medical staff, so as to improve the real professional value of medical staff.

We investigated whether FTY-720 might have an effect on bleomycin-induced pulmonary fibrosis through inhibiting TGF-β1 pathway, and up-regulating autophagy. The pulmonary fibrosis was induced by bleomycin. FTY-720 (1 mg/kg) drug was intraperitoneally injected into mice. Histological changes and inflammatory factors were observed, and EMT and autophagy protein markers were studied by immunohistochemistry and immunofluorescence. The effects of bleomycin on MLE-12 cells were detected by MTT assay and flow cytometry, and the related molecular mechanisms were studied by Western Blot. FTY-720 considerably attenuated bleomycin-induced disorganization of alveolar tissue, extracellular collagen deposition, and α-SMA and E-cadherin levels in mice. The levels of IL-1β, TNF-α, and IL-6 cytokines were attenuated in bronchoalveolar lavage fluid, as well as protein content and leukocyte count. COL1A1 and MMP9 protein expressions in lung tissue were significantly reduced. Additionally, FTY-720 treatment effectively inhibited the expressions of key proteins in TGF-β1/TAK1/P38MAPK pathway and regulated autophagy proteins. Similar results were additionally found in cellular assays with mouse alveolar epithelial cells. Our study provides proof for a new mechanism for FTY-720 to suppress pulmonary fibrosis. FTY-720 is also a target for treating pulmonary fibrosis.