Primary cell culture and morphological observation of bulbar conjunctival fibroblasts in conjunctivochalasis / 国际眼科杂志(Guoji Yanke Zazhi)

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.

Effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To observe the effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis. <p>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. <p>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 <i>P</i><0.05). After drug treatment, 7 cases(7 eyes)in artificial tears group and 4 cases(4 eyes)in combined treatment group of CCH patients were treated by operation. The positive rate of aging cells in combined treatment group was significantly lower than that in artificial tears group(16.00±7.84 <i>vs </i>39.00±14.09, <i>P</i>=0.013).<p>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.

Effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To observe the effects of Qi Jing Mingmu decoction combined with artificial tears on the clinical results and cell aging of conjunctivochalasis. <p>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. <p>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 <i>P</i><0.05). After drug treatment, 7 cases(7 eyes)in artificial tears group and 4 cases(4 eyes)in combined treatment group of CCH patients were treated by operation. The positive rate of aging cells in combined treatment group was significantly lower than that in artificial tears group(16.00±7.84 <i>vs </i>39.00±14.09, <i>P</i>=0.013).<p>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.

Evaluation of Subconjunctival Remnant Particles after High-frequency Radio-wave Electrosurgery for Conjunctivochalasis

PURPOSE: To investigate the clinical manifestations and properties of remnant particles in the subconjunctival space after high-frequency radio-wave electrosurgery for conjunctivochalasis. METHODS: We performed a retrospective, observational case series with in vitro experimental imaging in nine eyes from eight patients who presented with small dark-gray lesions during follow-up after high-frequency radio-wave electrosurgery for conjunctivochalasis. General examination including slit-lamp examination and visual acuity testing was performed preoperatively and postoperatively. During follow-up, we evaluated remnant particles and any other complications including granuloma or conjunctival injection with slit-lamp photography and anterior optical coherence tomography. Coagulation tips were investigated with scanning electron microscope and energy dispersive X-ray spectroscopy to analyze the insulating electrode and assess changes to tips after repeated use. RESULTS: None of the patients included in this study experienced any change in visual acuity or major complications postoperatively. Small dark-gray lesions (0.3 to 0.5 mm in size) were observed in the inferior bulbar sub-conjunctival space in the location where high-frequency radio-wave electrosurgery had been performed. Cirrus high-definition optical coherence tomography images revealed focal hyper-reflection with a posterior shadow, suggesting foreign particles. Scanning electron microscopy and energy dispersive X-ray spectroscopy imaging analysis revealed peaks of carbon and fluorine complexes, consistent with the polytetrafluoroethylene coating on the electrode. CONCLUSIONS: There were no instances of inflammatory reaction, particle migration, or major complications due to particles. Physicians should be aware of the possibility of remnant polytetrafluoroethylene particles in subconjunctival tissue when using insulated coagulation tips subjected to repeat sterilization.

Effect of Qijingmingmu decoction granule on MAPK signal pathway in fibroblasts of conjunctivochalasis stimulated by TNF-α / 眼科新进展

Objective To observe the effects of Qijingmingmu decoction granule on the expression of mitogen-activated protein linase (MAPK) signal pathway in fibroblasts of conjunctivochalasis (CCH) under the stimulation of tumor necrosis factor (TNF-α) and to explore the pathogenesis and effective treatment method of CCH.Methods CCH conjunctival fibroblasts were cultured in vitro and divided into CCH group,CCH + TNF-α group and CCH + TNF-α + Qijingmingmu decoction group.CCK-8 assay was used to determine the effective concentration of Qijingmingmu decoction granule,and 10-2 mg · L-1 TNF-α was added in the cultured fibroblasts of the latter two groups,followed by interference with Qijingmingmu decoction granule for 48h.The expression of MAPK signal pathway related protein and mRNA were detected by ELISA,Western Blot and RT-PCR.Then the results were statistically analyzed.Results CCK-8 assay showed the effective concentration of Qijingmingmu decoction granule was 1.61g · L-1.And there were significant differences in the total A values (A450) of extracellular regulated protein kinase (ERK),c-jun N-terminal kinase (JNK),p38 mitogen activated protein kinase (p38 MAPK) and their phosphorylation levels among the three groups (all P ＜ 0.05).Moreover,TNF-α could significantly up-regulate the expression of ERK1/2,JNK1/2,p-JNK1/2,p38 MAPK and p-p38 MAPK in CCH fibroblasts (all P ＜0.05),while Qijingmingmu decoction down-regulated their expressions in CCK fibroblasts after TNF-α stimulation (all P ＜ 0.05).Furthermore,the total differences in p-ERK1/2,p38 MAPK and p-p38 MAPK protein were significant in the three groups (all P＜0.05);and TNF-α could significantly up-regulate the expression of p-ERK1/2,p-JNK1/2,p38 MAPK and p-p38 MAPK protein in CCH fibroblasts (all P ＜0.05),while Qijingmingmu decoction down-regulated p-ERK1/2 and p38 MAPK expressions in CCK fibroblasts after TNF-α stimulation (both P ＜ 0.05).In addition,the total differences in ERK1/2 and p38 MAPK mRNA were significant in the three groups (both P ＜ 0.05);and TNF-α could significantly up-regulate the expression of ERK1/2 and p38 MAPK mRNA in CCH fibroblasts (both P ＜ 0.05),while Qijingmingmu decoction down-regulated p38 MAPK mRNA expressions in CCK fibroblasts after TNF-α stimulation (P ＜ 0.05).Conclusion The inflammatory factor TNF-α can up-regulate the expressions of MAPK signal pathway related protein and mRNA in CCH fibroblasts,resulting in the occurrence and development of CCH,and meanwhile Qijingmingmu decoction granule can downregulate the expression of MAPK signal pathway to play the therapeutic role in CCH to some extent.

Changes of tear film stability, goblet cell and mucin 5AC expression in conjunctivochalasis patients / 眼科新进展

Objective To observe the changes of tear film stability,goblet cell and mucin 5AC expression in conjunctivochalasis patients,and explore the mechanism of conjunctivochalasis.Methods Conjunctivochalasis patients (30 cases) and single age-related cataract patients (15 cases) were collected as conjunctivochalasis group and normal control group.Eye symptom assessment (OSDI score),tear break-up time (BUT),Schirmer Ⅰ test,tear fern crystallization tests were performed for all the selected persons.Conjunctival rescent-shaped resections were made for all the conjunctivochalasis patients.Conjunctival tissue samples were stained by HE staining,AB staining,mucin 5AC immunohistochemical staining from the conjunctivochalasis group and norral control group respectively,and then statistical analysis was made.Results The OSDI score in the conjunctivochalasis group (37.80 ± 8.94) was significantly higher than that in the normal control group (11.40 ±4.08) (P ＜0.01).BUT in the conjunctivochalasis group (6.70 ± 2.76) s was significantly lower than that in the normal control group (13.67 ± 3.48) s (P ＜ 0.01).Schirmer Ⅰ test in the conjunctivochalasis group (6.23 ± 3.13) mum was significantly lower than the normal control group (13.40 ± 3.74)mm (P ＜ 0.01).Tear ferbing crystallization of the conjunctivochalasis group was decreased significantly compared with the normal control group (x2 =14.309,P =0.003).Light microscopic showed that conjunctival thickness was thinned,collagen fibers were less,elastic fiber was reduced,the lamina propria and interstitial were congestion and edema,the number of goblet cells was significantly reduced,and the positive staining of mucin 5AC staining was significantly lower in the conjunctivochalasis group than in normal control group (x2 =9.499,P =0.023).Conclusion For patients with conjunctivochalasis,the tear film function is affected,goblet cells are decreased,tear fern crystallization is decreased,mucin 5AC content is decreased,which finally leads the excessive conjunctival relaxation and abnormal ocular surface and tear.

Inflammatory mechanism and significance of tumor necrosis factor-α and interleukin-1β in pathogenesis of conjunctivochalasis / 中华实验眼科杂志

Background Conjunctivochalasis (CCh) is a common age-related ocular surface diseases.Researches showed that the inflammatory factors are upregulated in conjunctiva and tear of CCh patients,inferring inflammation participates in the pathogenesis of CCh,however,its mechanism is unclear now.Studies also showed that the expression of matrix metalloproteinases (MMPs) in CCh conjunctiva is elevated.However,the association between inflammatory factors and MMPs is unknown.Objective This research was to observe the effects of tumor necrosisfactor-α (TNF-α) and interleukin-1 β (IL-1 β) on the expression of MMP-1,MMP-3,tumor necrosis factor-stimulatedgene-6(TSG-6) and pentraxin-3 (PTX3) in cultured CCh fibroblasts.Methods Conjunctival fibroblasts wereisolated and cultured from CCh-derived and cataract-derived human conjunctiva explants.The cells were treated using20 ng/ml PBS,TNF-α or IL-1β for 4 hours,respectively.The expression levels of MMP-1,MMP-3,TSG-6 and PTX3genes and proteins were detected by quantitative real time PCR and Western blot assay,respectively.Results Thesecond-generation cells showed a long and fusiform shape with ovoid nucleus and radial agreement,and the cellprocessors connected each other.The differences of the relative expression levels of MMP-1,MMP-3,TSG-6 and PTX3mRNA in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-αand IL-1 β (MMP-1 mRNA:Fgroup =2.611,P =0.116;Fi =161.564,P =0.000;MMP-3 mRNA:Fgroup =5.201,P =0.029;Fintervene =211.021,P =0.000;TSG-6 mRNA:Fgroup =47.209,P =0.000;Fintervene =119.340,P =0.000;PTX3 mRNA:Fgroup =40.512,P =0.000;Fintervene =93.935,P =0.000).Compared with the cataract group with PBS treatment,the expression levels of MMP-1,MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with PBS treatment or the cataract group with TNF-α and IL-1 β treatment (all at P＜0.05).Compared with the CCh group with PBS treatment,the expression levels of MMP-1,MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with TNF-α and IL-1β treatment (all at P ＜ 0.05).The differences of the relative expression levels of MMP-1,MMP-3,TSG-6 and PTX3 proteins in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-α and IL-1 β (MMP-1:Fgroup =84.702,P =0.000;Fint =48.900,P =0.000;MMP-3:Fgroup =112.818,P =0.000;Fint =194.980,P =0.000;TSG-6:Fgroup =56.867,P =0.000;Fint =70.356,P =0.000;PTX3:Fgroup =1.488,P =0.231;Fint =89.872,P =0.000),and the expression changes of MMP-1,MMP-3,TSG-6 and PTX3 proteins were coincident with the genes among the groups with various treatments.Conclusions The expressions of MMP-1,MMP-3,TSG-6 and PTX3 in conjunctival fibroblasts are upregulated in CCh eyes.The interaction of TNF-α and IL-1 β with MMPs is probably involved with the pathogenesis and development MMPs probably.

Study on the expression of mucin in conjunctival tissue and tear in eyes with conjunctivochalasis / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the expression of mucin in the conjunctival tissue and tear in eyes with conjunctivochalasis(CCh). <p>METHODS: Thirty-eight cases(38 eyes)of CCh were recruited as the CCh group and 36 cases(36 eyes)without CCh were included as the normal control group and the conjunctival tissue and tear were collected, respectively. Mucins(MUC2, MUC4, MUC5AC, MUC16)expression in conjunctival tissue were detected by immunohistochemical staining; Mucins(MUC2, MUC4, MUC5AC, MUC16)OD value in tear were determined by ELISA, the differences of which were compared between the CCh and normal control group. <p>RESULTS: No difference in the expression of MUC2 and MUC4 in conjunctival epithelia was observed between the two groups(<i>P</i>=0.315, <i>P</i>=0.156). There was significant difference in the expression of MUC5AC and MUC16 in conjunctival epithelia between the two groups(<i>P</i>=0.016, <i>P</i><0.01, respectively). No significant difference in the OD value of MUC2 in tear was found between the two groups(<i>P</i>=0.651). The OD value of MUC4 and MUC5AC in tear were significantly lower in CCh group that in the normal control group(<i>P</i><0.01, <i>P</i><0.01, respectively), so was the OD value of MUC16(<i>P</i>=0.022). <p>CONCLUSION: MUC5AC and MUC16 both decreased in the conjunctiva and tear of eyes with CCh. The further study may reveal the pathogenesis of conjunctivochalasis.

Conjuctivochalasis - a review.

Conjunctivochalasis is an ocular condition that is usually missed, unless specifically looked for. It is defined as a redundant loose non edematous inferior bulbar conjunctiva. Though generally asymptomatic, It may also present as dryness and foreign body sensation. Various grading systems like Mirmura ((based on Lid-parallel conjunctival folds), Mellers (based on lid-parallel conjunctival folds, punctual occlusion and conjunctival fold changes during down gaze and digital pressure) and Zhang’s based on conjunctival folds, dryness, foreign body sensations, epiphora symptoms, punctual occlusion, tear meniscus height, tear film break-up time (BUT), and conjunctival fold changes during down gaze) system has been outlined. Various etiological aspects like dissolution of the Tenon’s Capsule and role of expression of matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-3 (MMP-3) has been dealt in detail. Different surgical approaches like pastepinch- cut conjunctivoplasty and their steps are specified. Conjunctivochalasis has to be understood in a wider depth and more research work is required to understand CCh in better detail so that preventive steps can be implemented with regard to secondary causes.

Expressions of pentraxin-3 and tumor necrosis factor-α stimulated gene-6 proteins in fibroblasts of conjunctivochalasis patients / 中华实验眼科杂志

Background Conjunctivochalasis is an age-related inflammatory ocular surface disease manifesting redundant,loose conjunctiva folds.Studies showed that conjunctivochalasis often accompanies with inflammatory response,indicating that a variety of inflammatory cytokines are involved in pathogenesis of conjunctivochalasis.Objective This study attempted to investigate the expressions of pentraxin-3 (PTX-3) and tumor necrosis factor-α stimulated gene-6 (TSG-6) proteins in fibroblasts of conjunctivochalasis.Methods The protocol was approved by Ethic Committee of Putuo Hospital affiliated Shanghai University of Traditional Chinese Medicine.Thirteen eyes of 13 patients with conjunctivochalasis,16 eyes of 16 patients with age-related cataract and 16 eyes of 15 patients with pterygium were included in Putuo Hospital Affiliated to Shanghai University of Traditional Medicine from January,2011 to June,2012.The samples of conjunctival tissue were collected in the patients during surgery under the informed consent,and fibroblasts were cultured and passaged by explant culture method.Content of PTX-3 and TSG-6 proteins in cell supernatant was detected by ELISA.Results Translucent poor,dark black area was around the cell overflow.Cultured cells of third generation showed long shuttle type,uniform size,radiated out in all directions,and oval nuclei was sited in the cytoplasm,surrounded by varying the length of cross-linking of cell processes.The concentrations of PTX-3 protein in cell supernatant were (2 182.33 ± 738.68) pg/ml in the conjunctivochalasis group,which were higher than (738.32±335.00) pg/ml in the age-related cataract group and (981.37±416.04)pg/ml in the pterygium group,showing a significant difference among the three groups (F =6.474,P=0.032).The contents of TSG-6 proteins in cell supernatant were (59.10±6.52) pg/ml,(53.99± 11.16) pg/ml and (35.01±5.33)pg/ml in the conjunctivochalasis group,pterygium group and age-related cataract group,with a significant difference among the three groups (F =7.421,P =0.024),and contents of TSG-6 proteins in the agerelated cataract group was lowest,however,no significant difference was found in the TSG-6 contents between the conjunctivochalasis group and the pterygium group (P＞0.05).Conclusions Inflammatory reaction participates in the pathogenesis and development of conjunctivochalasis,up-regulation of PTX-3 and TSG-6 expression in fibroblasts might partakes in the pathogenesis of conjunctivochalasis.

Rapeutic effects analysis of Pranoprofen on tear dynamic abnormity induced by conjunctivochalasis / 国际眼科杂志(Guoji Yanke Zazhi)

? AlM: To investigate the therapeutic effects of Pranoprofen on tear dynamic abnormity induced by conjunctivochalasis ( CCh) . ?METHODS: Seventy cases ( 70 eyes ) whose the main symptoms was epiphora induced by CCh were divided into II, Ⅲ and Ⅳ in 3 groups according to Zhang’s grading standards, meanwhile, all the patients were treated with the eye drops of Pranoprofen for 2wk. After that, the curative effect of each group was analyzed before and after treatments according to the symptoms, dye disappearance test, bitter taste test and tear break-up time. ?RESULTS: After 2wk treatment of Pranoprofen, the symptom of patients at grade I and II improved obviously grade Ⅲ improved more, grade Ⅳ did little. The positive rate of dye disappearance was the highest in grade II, the second in grade Ⅲ, and the third in gradeⅣ. There was significant difference among the 3 groups (P0. 05). ?CONCLUSlON: Pranoprofen has good effects on tear dynamic abnormity induced by grade II of cch.

Changes in Area of Conjunctiva and Tear Meniscus Measured Using Anterior Segment Optical Coherence Tomography after Conjunctivochalasis Surgery

PURPOSE: To evaluate cross-sectional areas of conjunctiva and tear meniscus of conjunctivochalasis using Fourier-Domain RTVue-100 optical coherence tomography (OCT) before and after conjunctivochalasis surgery. METHODS: Thirty-one patients (33 eyes) with symptomatic conjunctivochalasis were recruited for this study between June 2013 and April 2014. All patients underwent crescent-shaped conjunctiva resection and amniotic membrane transplantation. Anterior segment OCT (AS-OCT) imaging was performed and tear break-up time was evaluated prior to and 3 months after the conjunctivochalasis surgery. Cross-sectional areas of conjunctiva and tear meniscus of conjunctivochalasis at 7 locations (1 center, 3 nasal and 3 temporal areas) were measured in all patients. RESULTS: The mean age of patients was 66.3 +/- 10.8 years. Cross-sectional areas of conjunctivochalasis at all locations significantly decreased from 0.487 +/- 0.42 mm2 to 0.007 +/- 0.011 mm2 (p < 0.001), whereas no significant changes in cross-sectional areas of tear meniscus at all 7 locations were observed after the surgery. Mean tear break-up time significantly increased from 2.26 +/- 0.69 sec to 3.81 +/- 1.22 sec following the surgery. CONCLUSIONS: Using AS-OCT, in this study we showed that areas of conjunctiva decreased and areas of tear meniscus were unchanged after conjunctivochalasis surgery.

Observation of limbal - stem - cell transplantation to treat pterygium combined with conjunctivochalasis / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To observe the efficacy, complications and ocular surface tear dynamics of the treatment for pterygium combined with conjunctivochalasis by inferior limbal-stem-cell transplantation. METHODS: All 30 patients ( 46 eyes ) diagnosed as pterygium combined with conjunctivochalasis in our hospital from December, 2010 to December, 2012 with a mean age of 61. 2y (SD 7. 2, range 49-75) were subjected to observation. After excision of nasal pterygium, an equivalent size of conjunctival flap linked with corneal limbal stem cell from inferior limbus was transplanted to the exposed sclera. Afterwards, the inferior bulbar conjunctiva was trimmed and fixed onto surface of sclera and inferior corneal limbus. After one-year follow-up, the comparisons regarding vision acuity between pre-operation and post-operation ( 1mo later ) as well as ocular-surface tear dynamics changes including several indexes like altitude and integrity of tear meniscus, break-up time ( BUT ) , corneal fluorescein staining and chloramphenicol gestation test before and 3mo after surgery were made. The operative complications and the recurrence rates 1y after operations were recorded. RESULTS: Median visual acuity was 0. 2 (0. 04-0. 6) before the operation and median visual acuity was 0. 4 (0-04-1. 0) 1mo after the operation. Of total 46 eyes in 21 eyes ( 45. 7%), visual acuity improved. No vision changes were found in other 25 eyes ( 54. 3%). Significant difference ( PCONCLUSION: Inferior - limbal - stem - cell transplantation to treat pterygium combined with conjunctivochalasis obtains better results. It proves to eradicate or relieve symptoms of patients effectively and ameliorate ocular-surface tear dynamics.

Effects of qijingmingmu soup on the expression of matrix metalloproteinases in the conjuntival fibroblasts of conjunctivochalasis / 中华实验眼科杂志

Background Our previous study determined that expressions of matrix metalloproteinases (MMPs) and tissue matrix metalloproteinase inhibitors (TIMPs)change in the conjuntival fibroblasts of conjunctivochalasis in vitro.To seek a suitable drug is very important in the prevention and treatment of conjunctivochalasis.Objective This study was to explore the effect of qijingmingmu soup on the expressions of MMPs and TIMPs in human conjunctival fibroblasts of conjunctivochalasis.Methods Twenty-four SD rats were randomized into two groups.Qijingmingmu soup was administration gastrically for consecutive 3 days,and normal saline solution was given in the same way in the control group.The blood was collected from aortaventralis and drug serum was prepared.Human conjunctival samples were obtained during the surgery of conjunctivochalasis relaxation and cultured in the DMEM containing 10％ fetal bovine serum,20％,15％,10％,5％ of drug serum and 8 ml/L epidermal growth factor(EGF) was added into the medium respectively.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expressions of MMP1,MMP3,TIMP1 and TIMP3in conjunctival fibroblasts.Results Cultured cells grew well with the fusiform shape and showed the positive response for vimentin.The expression value of MMP1 (A value)in the cells was declined after administration of qijingmingmu soup.A significant difference was found in the expression of MMP1 among the control group,20％,15％,10％,5％ drug serum groups and EGF group(F=466.664,P＜0.05),and that in the 10％ ([9.92±0.14] mg/L) and 20％ ([11.87 ±0.11] mg/L) drug serum groups was significantly lowed in comparison with the control group([16.31±0.10] mg/L)(t=99.974,87.394,P＜0.05).The expression value of the MMP3in the cells in the various drug serum groups,EGF group and the control group was significantly different(F=158.168,P＜0.05),with a lower value in the 20％ drug serum group compared with the control group ([3.50±0.03] mg/L vs.[4.44 ± 0.11] mg/L) (t =21.991,P ＜ 0.05).Also,the significantly different expressing value of TIMP1 was seen among all the groups (F=183.508,P＜0.05),and expressing value of TIMP1 in the 15％ drug serum group was(1.88±0.06)mg/L,which was lower than(3.20±0.32) mg/L of the control group(t=10.353,P＜0.05).Furthermore,the expressing value of the TIMP3 in the cells was significantly different among the various groups(F=54.503,P＜0.05),and that of the 20％ drug serum group was (1.743±0.065)mg/L and it was significantly higher than (1.54 ± 0.05) mg/L of the control group (t =5.046,P =0.004).However,the expressing value of TIMP3of the 15％,10％ and 5％ drug serum groups was lower than that of the control group,respectively all at(P＜0.05).Conclusions Qijingmingmu soup drug serum at the concentration of 20％ can down-regulate the expressions of MMP1,MMP3,TIMP1 and up-regulate the expression of TIMP3 in human conjunctivochalasis bulbar conjunctival fibroblastsin vitro,which probably plays preventive and therapeutic effects on conjunctivochalasis.

The Effects of Conjunctival Shield on Pain Alleviation During Cataract Surgery in Conjunctivochalasis Patients

PURPOSE: To evaluate the effect of conjunctival shield on pain alleviation during cataract surgery in conjunctivochalasis patients for conjunctiva drawn into speculum suction holes during cataract surgery. METHODS: Forty patients with conjunctivochalasis undergoing cataract surgery using un aspirating speculum were evaluated in the present study. A thin piece of surgical wipe (conjunctival shield) was inserted between the lid and bulbar conjunctiva to shield the conjunctiva from the suction hole and thereby prevent it from being sucked into the hole. Additionally, patients were asked to rank pain following speculum placement both before and after conjunctiva shield insertion by visual analogue scale. RESULTS: The pain scores were 2.50 +/- 0.78 points before conjunctival shield insertion, and 1.25 +/- 0.66 points after conjunctival shield insertion, which was a significant decrease (p = 0.01). In addition, the results showed a 70% reduction in pain following shield insertion. CONCLUSIONS: Conjunctivochalasis causes pain during cataract surgery when using an aspirating speculum. The conjunctival shield insertion is a safe and simple method for pain-free cataract surgery in conjunctivochalasis patients.

Histopathologic Characteristics of Conjunctivochalasis

PURPOSE: In this study we investigated the histopathologic characteristics of conjunctivochalasis and its association with lymphangiectasis using impression cytology and conjunctival excisional biopsy in patients with conjunctivochalasis. METHODS: This prospective study included 14 outpatients diagnosed with conjunctivochalasis, and who had excisional biopsies performed between March 1, 2012 and November 30, 2012. Preoperative slit-lamp examination, dry eye diagnostic test and impression cytology were also performed. For histological analysis, conjunctival sections were stained with hematoxylin and eosin (H&E), Verhoeff-van Gieson (VVG) elastic staining, and monoclonal antibody D2-40, a lymphatic endothelial marker. RESULTS: Tear break-up time and Schirmer's value were decreased in patients with conjunctivochalasis. Impression cytology showed a decrease in goblet cell density and an increase in nucleoplasmic/cytoplasmic ratio. The patients' conjunctival tissues exhibited an infiltration increase of chronic inflammatory cells on H&E stain, and a decrease in collagen density with degeneration of elastic fibers on VVG stain compared to tissues from the control subjects. D2-40 immunohistochemistry revealed the dilated subconjunctival lymphatics in patients with conjunctivochalasis. CONCLUSIONS: This study suggests that the characteristic histopathological features of conjunctivochalasis include dilated lymphatic vessels as well as decreased goblet cell and collagen densities with degeneration of elastic fibers.

Effect of Electrocauterization on the Inflammation of the Conjunctiva in Experimental Animal Model

PURPOSE: Recently, conjunctivochalasis repair surgery using electrocauterization has been gaining popularity. However, patients with electrocauterized conjunctivoplasty tend to complain of more postoperative pain than patients undergoing simple excision with suturing. Therefore, we investigated the effects of electrocauterization on inflammation of the conjunctiva using an experimental animal model and compared these with the effects of simple excision with suturing. METHODS: Ten New Zealand white rabbits underwent cauterization in the right eyes and excision and suturing in the left eyes. For each eye, we excised or electrocauterized the inferior bulbar conjunctiva, 1 mm in width and 6 mm in length, 2 mm from the limbus. A fine-needle electrode was inserted subconjunctivally, and electrocauterization was performed. In the contralateral eye, the corresponding area was excised and re-approximated with 10-0 nylon sutures. Sutures were removed after 14 days. Tissue samples were obtained at 21 days post-procedure, and inflammatory cells were counted in five randomly selected fields (x200) on hematoxylin-eosin stained slides. Tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta concentrations in tears were measured using enzyme linked immunosorbent assays. RESULTS: All cauterized eyes demonstrated smooth surface healing without scarring after 5 days, whereas sutured eyes presented with mild edema with some scarring until the suture was removed. The number of inflammatory cells was significantly greater in sutured eyes compared with cauterized eyes (p = 0.035, Mann-Whitney U-test) at 21 days post-procedure. Tear TNF-alpha and IL-1beta concentrations at 21 days were similar in both groups. CONCLUSIONS: Electrocauterization for conjunctivoplasty seems to be advantageous in terms of inflammation compared with simple suturing and excision.

Ultrastructure change of conjunctiva and fascia tissue of conjunctivochalasis / 中华实验眼科杂志

Background Bulbar conjunctiva tissue appears to be thinning,elasticity declined,tension reduced and fascia atrophied in conjunctivochalasis.Histopathological examination of conjunctivochalasis shows decrease of elastic fibers and melt of collagen fibers.But there are fewer studies on the ultrastructure of conjunctiva of conjunctivochalasis up to now.Objective This study was to investigate the ultrastructure change of conjunctiva tissue in conjunctivochalasis.Methods Five loose conjunctiva samples of conjunctivochalasis and 5 normal conjunctival tissue samples were collected and ultra-microstructure changes of these samples were observed under the transmission electron microscope.Results The number of fibroblasts in conjunctivochalasis lamina was progressively decreased.The shape of fibroblasts was long and fusiform.Somatic synapse was slim.Nucleus-cytoplasm ratio was increased.Disorder,scattered and broken of the collagen fibril were seen,and some areas were dissolved or lacunae.Normal conjunctival fibroblasts were oval or polygonal,with wieners and long somatic synapse,and intercellular matrix was full of collagen fibril and dense arranged fibers.Fibroblasts in fascia of eonjunctivochalasis were linear,and collagen fibril was seriously defected.Fascia fibroblasts of normal bulbar conjunctiva were spindleshaped and bigger than conjunctivochalasis fibroblasts.There were full of collagen fibrils in intercellular matrix.Conclusions The collagen fibril is decreased and fibroblast cells are degenerated in lamina and fascia of conjunctivochalasis.

The Effect of Conjunctivochalasis Surgery Using a High-Frequency Radio-Wave Electrosurgical Unit

PURPOSE: To evaluate the effects of conjunctivochalasis surgery using a high-frequency radio-wave electrosurgical unit. METHODS: Twenty-seven eyes of 14 patients with conjuctivochalasis who received surgeries with shrinkage of the inferior bulbar conjunctiva using a high-frequency radio-wave electrosurgical unit (Ellman surgitron(R)) were evaluated. Conjuctivochalasis grade, the ocular symptoms, Ocular Surface Disease Index (OSDI), tear film break-up time (BUT), Schirmer test, and corneal staining with fluorescein were measured preoperatively, at 3 months postoperatively, and analyzed prospectively. RESULTS: Three months after the surgery, all patients' conjunctivochalasis became grade 0, and the ocular symptoms, OSDI score, BUT, and corneal staining with fluorerscein were also improved. Additionally, there were no specific complications during the follow-up period. CONCLUSIONS: Conjunctivochalasis surgery using a high-frequency radio-wave electrosurgical unit produced an effective improvement in conjuctivochalasis grade and symptoms and appeared to be a simple and safe procedure.

Treatment of Epiphora in Patients with Conjunctivochalasis Using Conjunctival Fixation to the Sclera

PURPOSE: To report the effects and complications of conjunctival fixation to the sclera in conjunctivochalasis patients with inferior punctal occlusion. METHODS: The authors of the present study evaluated the degree of conjunctivochalasis and performed Fluorescein Dye Disappearance Test (FDT) in 15 eyes of 8 patients diagnosed with conjunctivochalasis with inferior punctal occlusion. Under topical anesthesia, the inferior bulbar conjunctiva was attached to the sclera with 3 8-0 vicryl stitches 8 mm posterior from the limbus. After surgery, the relief of symptoms, postoperative complications and improvement of conjunctivochalasis were observed. RESULTS: One week after the surgery, all 15 eyes achieved a subjective improvement of symptoms and the degree of conjunctivochalasis and FDT showed statistical difference after surgery (p = 0.000, 0.000, respectively). A complication occurred in 1 eye which was a retinal hemorrhage due to scleral puncture. CONCLUSIONS: Conjunctival fixation to the sclera could improve epiphora in conjunctivochalasis patients with inferior punctal occlusion. However, this procedure should be performed with caution.