Eye tissue engineering is a new direction of scientific research.It applies body tissues,cells or scaffolds with specific biological activities in vitro or in vivo to construct ocular tissues or organs in order to promote and improve the ocular physical functions by maintaining,repairing and inducing tissue regeneration.With the increase of aging population and average life expectancy,eye tissue engineering and regenerative medicine will become an important researching area of ophthalmology,and its relevant products and technologies can become a platform for future alternative medical equipment applications in ophthalmology.Greater efforts on the basic research and continuous innovation of eye tissue engineering technologies in China,including translational medicine from basic research to industrial production and medical applications,will significantly build up the foundation of eye tissue engineering industry in China and greatly improve its competitiveness in international medical marketing,and these research achivements will provide new power for the development of ophthalmology in China.

Background Herpes simplex virus (HSV) disciform stromal keratitis is a T cell-mediated delayed-type hypersensitivity of corneal stroma.The treatment of HSV disciform stromal keratitis is the combination of glucocorticoid and antiviral drug before.However,the therapy is limited for intolerantble patients to glucocorticoid.In addition,the adverse reactions following usage of glucocorticoid can not be ignored.FKS06 solution is proved to has the immunosuppressive and antiinflammatory acttions with less side effects than glucocorticoid drug.But the study on the application of FK506 solution in treatment of HSV disciform stromal keratitis is lack.Objective This prospective clinical trail was to evaluate the efficacy of topical tacrolimus for HSV disciform stromal keratitis and its safety.Methods A prospective study was performed on 18 consecutive cases (18 eyes) who were diagnosed as active HSV disciform stromal keratitis from June 2011 to June 2012 in Zhongshan Ophthalmic Center under the approval of the Ethic Committee of Zhongshan Ophthalmic Center and informed consent of the patients,including 3 patients with refractory to prior steroid eyedrops.All patients received a combination topical tacrolimus eyedrops 4 times per day with topical and systemic anti-viral therapy for mean (56.0±7.9) days.Corneal thickness by slit lamp biomicroscope,visual acuity,intraocular pressure,corneal fluorescein staining,Schirmer Ⅰ test (S Ⅰ t) and tear breakup time (BUT) were performed on the patients before and 1 month,2 months after treatment and at the drug withdrawal to assess the treating efficacy of FK506,and the adverse reactions of eyes were recorded.Results Corneal edema was alleviated in (8.3 ± 2.5) days after treatment in all the patients and returned to normality in (25.3±11.5) days.Corneal edema of 2 eyes with refractory to previous steroid eyedrops showed a slower recovery procedure than that of the others.Corneal fluorescence staining scores were 4 (6,2),1 (2,1),1 (1,0) and 0 (1,0) before and 1 month,2 months and at drug withdrawal,respectively,with a significant difference among the four time points (H=39.90,P＜0.001),and the scores were significantly lower after treatment than before (all at P＜0.01).The S Ⅰ t values were (6.78±1.90),(7.39±3.53),(8.06±2.92) and (8.11±3.05) mm/5 min,respectively in the 4 time points,without statistically significant difference among various time points (F =0.94,P =0.43).The BUT values were (0.39 ± 0.50),(1.11 ± 0.90),(2.00 ± 1.08) and (3.39 ± 0.92) seconds,respectively,showing a significant difference among the 4 time points (F=34.54,P＜0.01) and had the increased values after treatment of FK506.Recurrence was seen in 2 eyes in 2 months and 5 months after cessation of tacrolimus.However,the eyes were healed after a second round regimen.No elevation of intraocular pressure and secondary infection were found in the patients during the treatment and follow-up period.Conclusions The combination therapy of tacrolimus with antiviral drug is effective and safe for HSV disciform stromal keratitis by alleviating corneal edema and promoting the rehabilitation of tear film.

Objective To prepare a specific anti-lactoferrin single chain variable fragment(ScFv).Methods Anti-lactoferrin clones were screened from a 'naive' phage antibody library against the immobilized lactoferrin antigen,then the clones were transformed to the E.coli HB2151 to give soluble expression of antibody fragments.The culture supernatant containing ScFv was purified by immobilized metal affinity chromatography,and then determined with SDS-PAGE and ELISA.Results The results demonstrated that ScFvs were specific;they did not react with transferrin,lysozyme and bovine serum albumin in ELISA.The SDS-PAGE showed that the ScFvs had high purity through affinity chromatography and the molecular weight of them was about 32 kD.Conclusions The successful generation of the ScFvs against lactoferrin provides a basis for further study and clinical applications for dry eye and other ocular diseases.

AIM: This study was designed to investigate the apoptotic effect of doxycycline in THP-1 cells.METHODS: After differentiated by PMA, THP-1 cells were treated with doxycycline at different concentrations ranging from zero to 80 mg/L. The morphological changes of THP-1 cells were observed under light microscope. MTT assay were used to examine the effects of doxycycline on proliferation of THP-1 cells. Apoptotic THP-1 cells were measured by Annexin-V flow cytometry analysis and TdT-mediated dUTP nick end labeling assay.RESULTS: Treated with a certain concentration of doxycycline, differentiated THP-1 cells contracted and turn round, a number of cells were dead. MTT assay and positive Annexin V-FITC on cell membrane and TUNEL assay showed that doxycycline induced apoptosis in THP-1 cells in a dose-dependent manner.CONCLUSION: Doxycycline induces apoptosis in THP-1 cells in a dose-dependent manner.

BACKGROUND:Increasing studies have found that, percutaneous vertebroplasty and bone cement are characterized by few traumas, few bleeding, simple operation, rapid recovery, rapid pain relief, and effective efficacy. However, tumor recurrence, symptom improvement, bone cement leakage and other complications make people controversial. OBJECTIVE: To explore and summarize clinical outcomes and safety of percutaneous vertebroplasty and bone cement for treating symptomatic vertebral hemangioma. METHODS:Fifty-two cases patients of symptomatic vertebral hemangioma (including 21 male and 31 female, aged 16-63 years) were treated with percutaneous vertebroplasty and bone cement. The lesions were found at T5-T12 in 36 cases and at L1-L3 RESULTS AND CONCLUSION: Al 52 patients were folowed up for at least 6 months after the operation. The Visual Analogue Scale and Oswestry Disability Index were significantly increased at 1 day, 3 weeks and 6 months post-surgery, when compared to before surgery (P< 0.05); the modified Macnab evaluation was excelent in 40 cases, good in 7 cases, fair in 3 cases, and poor in 2 cases, and the excelent and good rate was 90%. Only one patient had recurrence at 6 months after surgery and one patient had the leakage of bone cement. No other complications were found. Percutaneous vertebroplasty and bone cement could effectively cure vertebral hemangioma and relieve the pain, which is a safe treatment. in 16 cases. Clinical outcomes such as Visual Analogue Scale, Oswestry Disability Index and modified Macnab criteria, as wel as complication before and after surgery were evaluated.

Objective To compare the clinical effect and safety of percutaneous vertebroplasty between unipedicular and bipedicular approach for patients with osteoporosis vertebral compressive fractures. Methods A retrospective study was carried out on 147 patients with single osteoporotic vertebral compressive fractures from 2008 to 2014. Ninety-five cases were treated by PVP via a unipedicular approach (unipedicular group) and 52 cases were treated by the bipedicular approach (bipedicular group). The operation time, Cobb, frequency of X-ray exposure,complications like bone cement and postoperative VAS score were compared. Results No significant differences on the VAS score, cobb and complications were found (P > 0.05). However, there were significant differences on the operation time and X-ray exposure frequency (P<0.05). Conclusions No significant differences on clinical effects were found between the unipedicular and the bipedicular approach to percutaneous vertebroplasty for osteoporosis vertebral compressive fractures. However, the unipedicular approach can shorten the operation time and reduce X-ray exposure.

To realize the remote operation of the slit lamp microscope for department of ophthalmology consultation, and visual display the real-time status of remote slit lamp microscope, a remote slit lamp microscope consultation system based on B/S structure is designed and implemented. Through framing the slit lamp microscope on the website system, the realtime acquisition and transmission of remote control and image data is realized. The three dimensional model of the slit lamp microscope is established and rendered on the web by using WebGL technology. The practical application results can well show the real-time interactive of the remote consultation system.
Internet ; Ophthalmology ; methods ; Remote Consultation ; Slit Lamp ; Technology

AIM: To determine the optimal protocol and condition in which macaca irus mesenchymal stem cells (MSCs) are induced to differentiate into neuron-like cells by cryptotanshinone in vitro. METHODS: MSCs from macaca irus bone marrow were generated in vitro and induced with cryptotanshinone. The morphological changes of MSCs were evaluated by microscope. The positive percentages of neurofilament (NF), neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression were measured by immunocytochemistry with ABC staining. RESULTS: The result showed that MSCs were positive for CD29, CD44, CD105, CD166, and negative for CD34, CD71, CD80 and CD86. After induced with cryptotanshinone, MSCs began to display neuronal morphologies, such as contracted multipolar cell body and formed extensive networks. The percentages of positive NSE, NF expression were 68.3%?3.5%, 70.3%?1.5%, respectively. CONCLUSION: Macaca irus MSCs could be induced to differentiate into neuron-like cells in vitro by cryptotanshinone and might be applied in cell transplantation and gene therapy in nervous system disorders.

AIM: To investigate the survival, migration and differentiation of human bone marrow mesenchymal stem cells (hMSC) in the middle cerebral artery occlusion (MACO) model and to observe the influence on motor function in the animal model. METHODS: hMSC with Hoeschst 33342 were injected into the animal model of MACO after Shenqiye induced for half an hour and their survival, migration, differentiation and the behavior changes in MACO rats were examined. RESULTS: hMSC had good homogeneousness and immunological reaction after implantation. The results showed that hMSC survived in rat brain for a long time over six weeks. As time going, hMSC were found in much more areas in the rat brain. Immunofluorescence staining suggested that implanted hMSC expressed human neuron specific enolase, neurofilament, and glial fibrillary acid protein. At the same time, improvements in abnormal behavior of MACO rats were observed. CONCLUSION: hMSC differentiate into neurons in the brain of rats, which means that hMSC is an ideal seed cells for the therapy of cerebral infarction.

OBJECTIVETo evaluate the expression of the type I growth factor receptor family [epidermal growth factor receptor (EGFR), ErbB2 and ErbB3] by the epithelial cells in pterygium.

METHODSImmunoflourescent staining and Western blotting were performed to detect the expression pattern and quantity of EGFR, ErbB2 and ErbB3 proteins in the epithelia of 15 patients with primary pterygium and 12 subjects with normal conjunctiva.

RESULTSIn immunofluorescent staining, the EGFR protein was present in the basal cells while the ErbB2 and ErbB3 were expressed by the superficial cells in normal conjunctival epithelium. Of the pterygium cases 15, 11 were stained by EGFR, ErbB2 and ErbB3 in the full thickness of the epithelium and showed stronger staining compared with the control group. Four of them showed a similar staining pattern to the normal conjunctiva group. The density of protein bands detected by Western blotting for all three growth factor receptors was consistent with the immunofluorescent staining. Compared with normal conjunctiva, stronger protein bands of these three receptors were found in all of the pterygium specimens, in which EGFR, ErbB2 and ErbB3 were expressed in the full thickness, as shown by immunofluorescent staining.

CONCLUSIONSThe increased expression of EGFR, ErbB2 and ErbB3 proteins was present in pterygium, which indicated that pterygium is a disorder with abnormal proliferation. The abnormal expression of EGFR, ErbB2 and ErbB3 by the epithelium and the communication with cytokines in the stroma in pterygium may be a key pathogenic factor in this disorder.

Adult ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Pterygium ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Receptor, ErbB-2 ; metabolism ; Receptor, ErbB-3 ; metabolism