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AIM: To observe the growth status and morphological changes of primary cultured bulbar conjunctival fibroblasts in different stages of conjunctivochalasis(CCH), and to determine the best passage time, so as to obtain stable and consistent CCH bulbar conjunctival fibroblasts.METHODS: CCH primary bulbar conjunctival fibroblasts were obtained by tissue block adhesion method. The fibroblasts were purified by trypsin differential digestion method. The growth status and morphological changes of fibroblasts in different periods were observed and recorded under inverted microscope. The fibroblasts were identified by immunofluorescence cytochemical staining.RESULTS: After 24h of CCH conjunctival tissue adherent to the wall, a small number of cells would be seen crawling out around the tissue blocks. The logarithmic phase of cell growth was from the 2-7d. The cells grew fast, with vigorously proliferation, clear outline, uniform distribution, increas in numbers and clear nuclei. From the 9-15d, the cell growth entered the plateau stage, the tissue blocks gradually aged and lost activity. The cells grew slowly, arranged loosely, the volume became larger, the shape became flat, and a large number of granular substances and vesicles were produced in the cytoplasm. Some cells fell off from the bottom of the culture bottle, and large gaps appeared between the cells. After subculture and purification, the size and morphology of fibroblasts were basically the same. Through cell identification, fibroblasts were long spindle, flat star or multi-process spindle, wide in the middle, oval nucleus, relatively small at both ends, with 2-3 slender processes of different lengths extending outward.CONCLUSION: Primary CCH bulbar conjunctival fibroblasts can be successfully obtained by tissue block adhesion method. When the cells grow to the 8d, they can be digested and passaged to obtain stable and consistent CCH conjunctival fibroblasts.

AIM: To observe the effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis. METHODS: Forty cases(80 eyes)of grade II-Ⅳ CCH with liver-kidney Yin deficiency were randomly divided into two groups: combined treatment group and artificial tears group, which were treated with Qi Jing Mingmu decoction combined with artificial tears and simple artificial tears respectively. The international ocular surface disease index(OSDI), tear break-up time(BUT)and Schirmer Ⅰ test(SⅠt)were observed and the clinical effects were compared after 3mo treatment. For CCH patients with grade III or above, followed up for 3mo or more and willing to operate, the loose conjunctival tissue was removed and the cell aging related β-gal staining was performed on frozen sections. The results were statistically analyzed. RESULTS: The OSDI score(14.53±2.68), BUT 9.25±3.02s and SⅠt(8.95±3.57mm/5min)of combined treatment group were significantly better than those of artificial tears group after 3mo treatment(all Pvs 39.00±14.09, P=0.013).CONCLUSION: Qi Jing Mingmu decoction combined with artificial tears to treat CCH is more effective than simple artificial tears in relieving ocular symptoms, improving tear film and promoting tear secretion. Combined treatment can also reduce the cell aging in CCH, which can be used as a safe and effective treatment method in addition to surgical operation.

This study investigated the potential role of MAPK signaling pathways in conjunctivochalasis (CCH). Twenty loose conjunctival biopsy samples from 20 CCH and 15 conjunctival biopsy samples from 15 normal controls (CON) were collected. The conjunctival fibroblasts were cultured in vitro. Immunofluorescence, ELISA, Western blot and reverse transcription-polymerase chain reaction (RT-PCR) were used. Our results showed that the expression of p-ERK, p-JNK, and p-p38 in CCH conjunctiva was significantly higher than that in CON group. The expression of p38 MAPK, JNK, and ERK proteins in CCH fibroblasts was significantly higher than that in CON group. The total expression of MAPK mRNA in CCH fibroblasts was significantly higher than that in CON group. The activated forms of p38 MAPK, JNK, and ERK proteins and mRNAs might up-regulate the expression of MMPs in CCH loose conjunctival tissue and fibroblasts, causing the degradation of collagen fibers and elastic fibers and promoting the occurrence of CCH. Our results deepen the understanding of CCH pathological mechanism.

AIM: To measure the tear film thickness when with a good fitted night wear orthokeratology lens in youth, and observe the change of corneal parameters. METHODS: From July 2015 to August 2016, 51 eyes of 26 teenagers with myopia who would wear orthokeratology and accept regular follow-up were enrolled. The mean spherical equivalent refraction of teenagers was -2.78D±1.53D(-1.00D to-6.00D)before wearing orthokeratology, and the mean astigmatism was 1.29D±0.61D(0 to -2.68D). The tear film thickness were measured by optical coherence tomography(OCT)when the eyes wearing orthokeratology for 1mo. The surface regularity index(SRI)value was measured by corneal topography to compare its relationship with the tear film thickness and the changes of corneal anterior surface parameters during the period of wearing lens were observed. RESULTS: At 1mo after wearing orthokeratology, the mean tear film thickness of right eyes was 230.09±10.40μm, and that was 224.38±10.57μm of left eyes. The mean tear film thickness was 228.75±9.66μm in male eyes and that was 224.80±6.74μm in female eyes. The mean tear film thickness of all eyes was 227.05±10.29μm, and the mean SRI value was 0.16±0.14. There was no correlation between tear film thickness and SRI value(P>0.05). The FK value, SK value and average K value from before wearing to after wearing for 2d and 1wk gradually decreased, the difference was statistically significant(PP>0.05). CONCLUSION: OCT can be used as a safe and reliable method for measuring the thickness of the tear film space during the wearing of orthokeratology, which may be of certain value for the safety assessment of the long term wearing of orthokeratology. Orthokeratology works very quickly, corneal curvature decreases rapidly 2d after wearing and stabilizes during the period.

AIM:To compare the similarities and differences of central corneal thickness(CCT)measured by different devices, so as to provide a safe, effective, simple and accurate method for measuring corneal thickness. METHODS: Totally 95 eyes of 95 cataract patients enrolled continuously for preoperative examination in ophthalmology department of Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from September 2016 to December 2016. Handheld automatic PachPen ultrasonic pachymetry(USP), anterior segment optical coherence tomography(AS-OCT)and specular microscopy were respectively used to measure the central corneal thickness, and intraocular pressure was measured at the same time. The similarities and differences of these three different devices to measure the central corneal thickness and their correlations with intraocular pressure were observed. Then the data were analyzed statistically. RESULTS: The CCT value measured by PachPen ultrasonic pachymetry was 544.43±36.61μm, the CCT value measured by AS-OCT was 527.09±35.54μm, and the CCT value measured by specular microscopy was 533.20±30.17μm. There was significant difference between these three groups(F=6.272, P=0.002), and the CCT value measured by PachPen ultrasonic pachymetry was significantly higher than the other two groups. The correlation coefficients between the CCT values of these three groups and intraocular pressure were 0.290, 0.277 and 0.204(PPCONCLUSION: The CCT value measured by PachPen ultrasonic pachymetry is the highest. The second CCT value is measured by specular microscopy, and the smallest CCT value is measured by AS-OCT. Obvious correlation and good consistency were found in the CCT values measured by three types of devices, and the CCT values were all correlated with intraocular pressure. Therefore, we should take more attention in clinic.

Dry eye is a common ocular surface disease, which seriously affects the work and quality of life of the patients. In recent years, physical therapies for dry eye have developed rapidly. This paper summarizes the traditional physical therapies such as fumigation, atomization, eye massage, hot compress, moisture chamber glasses and so on, and bandage contact lenses and scleral contact lenses, intense pulsed light treatment, Blephasteam wet chamber warming device, LipiFlow heat pulse therapy and other advanced physical treatment methods widely used in recent years. We review the recent research progress about physical therapies for dry eye to provide reference for clinical treatment.

AIM: To investigate the expression of matrix metalloproteinases 1 and 3 (MMP-1 and MMP-3) and their tissue inhibitors of metalloproteinases 1 and 3 (TIMP-1 and TIMP-3) in the conjunctiva of eyes with conjunctivochalasis (CCh). METHODS: The conjunctival tissue was obtained from the CCh patients and controls, the MMPs/TIMPs expression concentration was determined by enzyme-linked immuno-sorbent assay (ELISA) and immunofluorescence staining. The expression levels of MMPs/TIMPs in the CCh fibro-blasts were determined by analyzing its concentration in the cellular supernatant that was abstracted from the in vitro cultured CCh fibroblasts. RESULTS: MMP-1 and MMP-3 levels determined by ELISA were both significantly higher in the CCh group than that in the control group (P=0.042, 0.022, respectively), so was the levels of TIMP-1 (P=0.010). No significant difference in the expression of TIMP-3 in conjunctiva was found between the two groups (P=0.298). The expression of MMP-1 and MMP-3 were both up-regulated significantly in the CCh group (P=0.040, 0.001, respectively) on immuno-fluorescence staining. MMP-1 and MMP-3 expression in the fibroblasts were both significantly higher in the CCh group than that in the control group (P=0.027, 0.001, respectively), while neither the TIMP-1 nor TIMP-3 expression was significantly different between the two groups (P=0.421, 0.237, respectively). CONCLUSION: The overexpression of MMP-1 and MMP-3 in conjunctival tissue and fibroblasts may play an important role in the pathogenesis and development of CCh.

Collagen protein has special biological characteristics and physical properties, including quaternary structure, low immunogenicity, biocompatibility, biodegradability and hemostatic properties and so on, which are the foundation of collagen protein being widely used and it has important relationship with the occurrence and development of some eye diseases, such as keratoconus, conjunctivochalasis, high myopia, etc. This paper reviews the recent progress of collagen protein in eye diseases.

AIM: To observe thickness and morphological changes of bulbar conjunctiva pre- and post epidemic hemorrhagic conjunctivitis(EHC)therapy by optical coherence tomography(OCT).METHODS: Observed morphological changes and measured the bulbar conjunctiva thicknesses of 29 cases(36 eyes)of incipient(1-2d)EHC patients, who were received and treated by department of ophthalmology, the Putuo Affiliated Hospital of Shanghai University of Traditional Chinese Medicine from May 2013 to December 2013, by OCT. Then measured the thickness again on 7, 14d after the therapy. RESULTS: Among 29 patients(36 eyes), 7d after the EHC therapy, in 27 cured eyes, the full-thickness(before 344.00±59.91μm, after 230.19±22.16μm, t=11.75, Pt=4.61, Pt=10.69, Pt=14.45, Pt=10.06, Pt=13.11, PCONCLUSION: The OCT is able to measure the thickness of bulbar conjunctiva in EHC patients. An significant increase was found in full, epithelial and stromal thickness of EHC patients' bulbar conjunctiva. With recovery from the disease, subepithelial fluid, interlaminar fluid and edema of the bulbar conjunctival stroma faded away firstly, which provide references for clinical therapies of the EHC.