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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.
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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.
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·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.
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@#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.
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To investigate the distribution of traditional Chinese medicine (TCM) Syndromes of diabetic retinopathy and explore the correlation between various symptoms. Based on the literature in databases of China Journal Full-text Database (CNKI), Wanfang database, VIP network, China biomedical literature database (CBM) and PubMed, SPSS 20.0 and IBM SPSS Modeler 14.1 software were used to analyze the location of disease, symptoms, tongue and pulse, and syndrome type distribution through frequency statistics. In addition, association rule algorithm was used to explore the basic rules for underlying symptoms combinations of diabetic retinopathy. A total of 560 articles were retrieved, and a total of 240 articles met the inclusion and exclusion criteria. In these documents, 62 types of TCM syndromes were found, involving 9 820 cases, 201 symptoms, 29 tongue conditions, and 36 pulses conditions; the first two locations for the disease were liver and kidney. Then the association rule analysis of high frequency symptoms was used to dig out 15 groups of latent syndrome, and 3 underlying symptom combinations among high frequency symptoms, tongue conditions and pulse conditions. The results of the study showed that Qi and yin deficiency was most common for the diabetic retinopathy, and the location of the disease was closely related to liver and kidney. In addition, these high-frequency symptoms and tongue conditions, pulse conditions, and underlying symptom combinations can occur as main symptoms at diagnosis, providing reference for us to study the epidemiology of PRO scale of the disease. They can also increase the weight of these symptoms directly as the main symptoms, which can be also used as an alternative entry pool for TCM syndrome diagnostic scale, laying foundation for the construction and optimization of TCM symptom database of diabetic retinopathy.
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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, 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.
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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.
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<p><b>BACKGROUND</b>Different diagnostic and grading systems of conjunctivochalasis have resulted in apparent disparity between the prevalence rates of recent population-based studies. This study aimed to investigate the disparity between 4-level system cited from Meller and Tseng in 1998 (abbreviated here as Meller's system) and 5-level system modified from Meller's system cited from Zhang and associates (abbreviated here as Zhang's system) regarding the diagnosis and the patients' preferences for the treatment of conjunctivochalasis in the general population.</p><p><b>METHODS</b>A total of 546 senile residents living in the Guiyangyuan community of Shanghai, China, participated in the study. The diagnostic criteria for conjunctivochalasis were based on two diagnostic grading systems: Meller's system and Zhang's system, which was modified from Meller's system. The participants' preference regarding medical treatment for conjunctivochalasis was determined according to the response to a question. One year later, a follow-up interview determines whether the patient had undergone surgery for conjunctivochalasis.</p><p><b>RESULTS</b>With Meller's system, 398 participants were confirmed as having conjunctivochalasis, and the prevalence rate was 72.89%. According to Zhang's system, only 213 participants were diagnosed as having conjunctivochalasis, and the prevalence rate was 39.01%. A total of 109 eyes underwent medical treatment or surgery for conjunctivochalasis in the following year, including eight eyes that were diagnosed as grade II and 101 eyes that were diagnosed as grade III according to Meller's system and five eyes that were diagnosed as grade I, 55 eyes that were diagnosed as grade II, 31 eyes that were diagnosed as grade III, and 18 eyes that were diagnosed as grade IV according to Zhang' system.</p><p><b>CONCLUSION</b>Diagnoses of conjunctivochalasis using Zhang's system are more consistent with patient requests and the medical treatment strategies used than diagnoses made using Meller's system.</p>
Subject(s)
Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Conjunctival Diseases , Diagnosis , Epidemiology , PathologyABSTRACT
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.
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<p><b>BACKGROUND</b>The crescent excision of the inferior bulbar conjunctiva has been advised as a surgical procedure in the management of conjunctivochalasis refractory to medical treatments. However, it is difficult for this procedure to design how much conjunctival tissue should be excised. This study aimed to present a quantitative locator for conjunctiva resection and evaluate its effect on the treatment of conjunctivochalasis (CCh).</p><p><b>METHODS</b>Poly β-hydroxyethyl methacrylate resin/β-hydroxyethyl methacrylate (HEMA, water gel) was used as the material to make the quantitative locator which was designed to suit the specific patient. Forty-six patients with bilateral symptomatic CCh were included in this prospective study. Of the patients, while the right eye underwent the popularly used crescent-shaped conjunctiva resection (group I), the left eye was treated with conjunctiva resection assisted by the quantitative locator (group II). International Ocular Surface Disease Index (OSDI), scores of remnant conjunctiva fold, complications and conjunctival cut healing, height of tear meniscus, tear break-up time (BUT), and time of surgery were evaluated. Tasting chloromycetin test (TCT) was used to evaluate how the lacrimal duct worked.</p><p><b>RESULTS</b>OSDI in group II (8.82 ± 2.36) was significantly lower than that in group I (14.67 ± 2.21) (t = 12.22, P < 0.01). The amount of conjunctiva fold remaining in group II was less than that in group I. Scores of remnant conjunctiva fold in group I were significantly higher than those in group II (t = 31.85, P < 0.01). While evaluation scores of conjunctival cut healing in group I were lower than those in group II, scores of complication in group I were significantly higher than those in group II at 8 weeks after surgery (t = 89.60, P < 0.01). There was no significant difference in eyes with normal BUT (χ(2) = 0.031, P = 0.985) between the two groups, as the case was in eyes with positive TCT (χ(2) = 0.14, P = 0.930) and in eyes with normal height of tear meniscus (χ(2) = 0.48, P = 0.780). Mean surgery time in group II ((17.11 ± 2.08) minutes) was significantly shorter than that in group I ((25.22 ± 4.78) minutes) (t = 13.84, P < 0.01).</p><p><b>CONCLUSION</b>A quantitative locator can be used as an effective, safe, and less time-consuming instrument to facilitate conjunctival excision for symptomatic CCh treatment.</p>
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Aged , Female , Humans , Male , Middle Aged , Conjunctiva , General Surgery , Conjunctival Diseases , General Surgery , Ophthalmologic Surgical Procedures , Methods , Prospective Studies , Treatment OutcomeABSTRACT
Background Researches showed that the incidence rate of cataract is high in the nickel mining area. Nickel sulphate can apparently inhibit the metabolism and proliferation of human lens epithelium cells. But the study on the injury mechanism of nickel on lens is still seldom. Objective Present study was to investigate the effect of nickel sulphate on the lens of SD rats. Methods Forty-five SPF SD rats aged from 7 to 14 days were grouped randomly into subcutaneous injection group, intraperitoneal injection group and blank group. Nickel sulphate of 2 g/L ( 10 mg/kg) was subcutaneously or intraperitonealy injected for 45 days. The opacity of rat lens was examined under the slit lamp at two-week interval and scored based on the criteria of LOCS II and LOCS III. The rats were sacrificed in 45 days after experiment and the lens were obtained for the pathological examination. Result The mean score of the anterior subcapsule opacity of rat lens was obviously higher in subcutaneous injection group compared with blank control group with a significant difference between them (t= 14. 311, P < 0. 05 ) , but no significant difference in the anterior subcapsule opacity between intraperitoneal injection group and blank control group (t = 4. 355 , P>0. 05 ). The score of posterior subcapsule opacity of lens were evidently higher in both subcutaneous injection group and intraperitoneal injection group than the blank control group (t = 9. 316,P = 0. 004;t = 7. 464, P = 0. 009) ,so was the mean score of the anterior +posterior subcapsule opacities(t = 23. 387,P=0. 000;t= 10. 533,P = 0. 002) and the total score of rat lens opacity ( t = 12. 358 , P = 0. 001; t = 10. 188 , P = 0. 003 ) . No significant differences were found in cortex opacity score and nuclear opacity score among three groups ( P > 0.05 ). Histopathology examination revealed that the degeneration of lens collagen protein was more serious in subcutaneous injection group and intraperitoneal injection group than the blank control group,and the injury degree of lens collagen protein was more dominant in subcutaneous injection group. Conclusion System administration of nickel sulphate induced the injury of anterior and posterior subcapsule of lens in SD rat.
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Objective To observe the protective effect of Erigeron Breviscapus(Vant.)Hand-Mazz(EBHM) on retinal ganglion cells(RGCs) and optic nerve in rabbits with elevated intraocular pressure(IOP). Methods Twenty rabbits with chronic elevated IOP were divided into two groups randomly: EBHM treated group(the ocular hypertension with EBHM treated subgroup and the normal IOP with EBHM treated subgroup) and untreated group(the simple ocular hypertension subgroup and the simple normal IOP subgroup). EBHM was irrigated into the stomachs to the treated group after IOP elevated continuously for 7 d.For light and electron microscopy studies,the rabbits' eyes were made into eyeball samples and optic nerve samples after 60 d.The density of RGCs,thickness of retinal nerve fiber layer(RNFL),and optic nerve axons were observed and quantitated by computer image analysis system.The ultra-microstructure changes of RGCs and optic nerve axons were observed by electron microscope.Results(①The RNFL) thickness,RGCs density and number of axons of the two ocular hypertension subgroups were decreased,compared with those of the two normal IOP subgroups(P