Objective To explore the change of gene expression of stress activated protein kinase (SAPK) and its upstream signalregulated molecule ——mitogen activated protein kinases(MAPKs) (MKK4 and MKK7) in hypertrophic scar and autocontrol normal skin. Methods The total RNA was isolated from 8 hypertrophic scars and 8 auto-control skin, and then mRNA was purified. The gene expressions of MKK4, MKK7 and SAPK were examined with reverse transcriptionpolymerase chain reaction(RT-PCR) method. Results In hypertrophic scar, both MKK7 and SAPK genes weakly expressed. In auto-control skin, the expression of these 2 genes was significantly elevated in comparison with hypertrophic scar (Plt;0.01). The expression levelsof these 2 genes were 1.5 times and 2.6 times as long as those of hypertrophic scar, respectively. Gene expression of MKK4 had no significant difference between autocontrol skin and hypertrophic scar (Pgt;0.05). Conclusion Decreased gene expression of MKK7 and SAPK which results in reducing cell apoptosis might be one of the mechanisms for controlling the formation of hypertrophic scar.
Objective To investigate the effects of genistein with different concentration on proliferation and apoptosis of cultured human retinal pigment epithelial (RPE) cells in vitro. Methods The effect of genistein with the concentration of 5，10，25，50，75，and 100 mg·L-1on the proliferation of cultured RPE was examined by tetrazolium salt (MTT) assay and AgNORs staining. The cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling (TUNEL) methods, in the mean time, the morphologic changes of cell apoptosis were observed by light microscopy and transmission electronic microscopy, the results of which were compared with the normal RPE cells. Results Genistein with the concentration of 25, 50, 75, and 100 mg·L-1had a dose-dependent and time-dependent antiproliferative effects on RPE cells with the inhibitory rate of 12.0%-64.6% (P＜0.05). The results of AgNORs staining showed that the number of AgNORs in the nucleolus decreased when treated by genistein. In TUNEL staining, the median of percent of apoptotic RPE cells was 7.6%, 9.8%, 13.7% when treated with 50 mg·L-1genistein, 10.3%, 16.4%, 23.4% when treated with 75 mg·L-1genistein, and 15.4%, 21.2%, 35.8% when treated with 100 m g·L-1genistein respectively for 24, 48, and 72 hours. After the treatme nt with 50 mg·L-1genistein for 48 hours, the apoptosis in the nucleolus of RPE cells was found. Conclusions Genistein with different concentrations has a dose-dependent and time-dependent antiproliferative effect on RPE cells. Genistein can induce the apoptosis of RPE cells when it reaches a certain extent of concentration. (Chin J Ocul Fundus Dis，2004，20：241-244)
Objective To observe the effect of gefitinib on expression of epidermal growth factor receptor (EGFR) in bile duct epithelial cells, and the feasibility of inhibiting hyperplasia of bile duct epithelial cells with gefitinib. Methods Sixty-one patients with hepatolithiasis having to be in hospital for surgery from the First People’s Hospital of Shuangliu county were selected, with 25-65 years old, average 46.92 years. The patients were randomly divided into therapy group and control group. There were 30 cases in therapy group, in which fine duct was placed on lesion bile duct during operation, and through whom gefitinib solution was perfused after operation. There were 31 cases in control group with only T tube drainage after operation. The bile duct sample was obtained respectively during the operation and 6 weeks and 12 weeks after operation. The histology and expression change of EGFR were observed by HE staining, immunohistochemistry and RT-PCR method respectively. Results There were no significant differences in pathohistology changes of bile duct and the EGFR protein and mRNA expression between therapy group and control group during operation. The hyperplasia of epithelium mucosae and submucosal gland in the therapy group were obviously decreased as compared with those in control group, the EGFR mRNA and protein expression in therapy group were weaker than those of control group （Plt;0.05） 6 weeks and 12 weeks after gefitinib treatment. Conclusion EGFR is overexpressed in the chronic proliferative cholangitis, and continuously local application of gefitinib after operation can specifically interrupt the activation and expression of EFGR and then effectively inhibit the hyperplasia of bile duct epithelial cells.
To determine the state of fibroblast during the process of development of hypertrophic scar (HS), 40 specimens of HS in different periods were collected. The expressions of prolifrating cell nuclear antigen (PCNA) and Ag-protein in nucleolar organizer regions (Ag NORs) as well as the content of total amino acids in the tissues were examined. The hypertrophic scar of 1st and 3rd month old, the expression of PCND and Ag NORs were the highest. In the 9th and 12th month old, althrough PCNA was nearly negative, but the expression of Ag NORs was low. The content of total amino acid was increased gradually as HS developed but the increase of amount of hydroxyproline was markedly slowed down in 9 month old HS. It was suggested that: (1) in the developing process of HS the proecess of overproliferation of fibroblasts was short and limitted in 1-3 months period in the process of wound lealing; (2) the synthesis of collagen was nearly stopped at 6 months, but that of other extracellular matrix such as fibronectin and proteoglycan might be continued to aggregate after 12 months.
Objective To evaluate the effect of vascular endothelial cell growth factor (VEGF) antisense oligodeoxynucleotides (ASODNs) on the expression of VEGF in rats with oxygen-induced retinopathy. Methods Thirty newborn Sprague-Dawley (SD) rats were randomly divided into 3 groups:normal control group, disposal group and non-disposed group, The animal models with oxygen-induced proliferative retinopathy were established by raising the rats in hyperoxic environment. Retrobulbar injection was performed with VEGF ASODNs or normal saline on the rats in 3 groups respectively. The intraocular tissues (all the tissues except the cornea, sclera, and lens) and serum were collected, and the expressions of VEGF were determined by using competitive enzyme immunoassay.Results The expressions of VEGF in intraocular tissues of rats in disposal group were significantly lower than those in non-disposed group (P＜0.05), and there was no statistical difference between the disposal and normal control group (P＞0.05). There was no significant difference of the expressions of VEGF in serum of rats between the disposal and non-disposed group (P＞0.05), which were both lower than those in the normal control group (P＜0.05). Conclusion VEGF ASODNs could significantly inhibit the expression of VEGF in intraocular tissues. (Chin J Ocul Fundus Dis，2003，19：172-174)
Objective To investigate the effect of hypericin on the activity of protein kinase C (PKC) in cultured human retinal pigment epithelium (RPE) cells in vitro.Methods RPE cells were cultured in standard medium with 10% serum concentrations containing 0.5 to 5.0 μmol/L hypericin with or without preincubation of phorbol 12-myristate 13-acetate (PMA). The activities of cytosolic PKC (c-PKC) and membranous PKC (m-PKC) were assayed by PKC kit. Results The original activities of c-PKC and m-PKC of RPE cells were (35.34±4.10) pmol·min-1·mg-1and (62.52±8.80) pmol·min-1·mg-1.The activity of c-PKC in RPE cells with PMA preincubation decreased rapidly in 5 minutes, with a subsequent slow decrease after 20 minutes and a decrease to 18% of the activity of c-PKC in RPE cells without PMA preinubation after 60 minutes. While the activity of m-PKC in RPE cells with PMA preincubation increased gradually after 5 minutes and reduced after reached the peak at 40 minutes, and then returned to baseline after 60 minutes, eventually decreased below 30% of the control group. When RPE cells were cultured with PMA for 48 hours, the activities of c-PKC and m-PKC were hardly detectable, while RPE cells were cultured with both PMA and hypericin, hypericin could counteract most of down-regulation by PMA. Conclusion Hypericin may inhibit the translocation of PKC in RPE cells,change the activity of PKC, promote the apoptosis of RPE cells likely,and then prevent proliferative vitreoretinopathy. (Chin J Ocul Fundus Dis，2003，19：55-58)
Proliferative vitreoretinopathy (PVR) is a common complication and major cause of blindness of ocular trauma. Many cytokines, including vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF), participate in the process of the pathogenesis of traumatic PVR. VEGF competitively inhibits binding of PDGF to its receptor (PDGFRα), enables indirect activation of PDGFRα by non-PDGF ligands, resulting in reduced p53 expression, cell proliferation and migration, which is a key point in the pathogenesis of traumatic PVR.
Objective To investigate the inhibitive effect of E2F decoy oligodeoxynucleotides (E2F decoy ODNs) on cultured human retinal pigment epithelial (HRPE) cells.Methods E2F decoy ODNs or scramble decoy ODNs at varied concentrations were put into the HRPE cells mediated by lipofectamineTM2000. The proliferative activity of HRPE was detected by methythiazolyl-terazollium assay, and the competitive combinative activity of E2F decoy ODNs and transcription factor E2F was detected by electrophoresis mobility-shift assay. Results The proliferation of HRPE was inhibited markedly by E2F decoy ODNs at the concentration of 0.2 μmol/L (P=0.002) in a dose-dependent manner but not by scrambled decoy. The results of electrophoresis mobility-shift assay showed that the combinative activity of transcription factor E2F was abolished completely by E2F decoy ODNs. Conclusions E2F decoy ODNs may sequence-specifically inhibit the combinative activity of transcripti on factor E2F,and inhibit the proliferation of HRPE cells.(Chin J Ocul Fundus Dis，2004，20：182-185)
Objective To observe the therapeutic effect of vitrectomy on Eales disease and the correlated factors affecting the visual prognosis and disease outcomes. Methods The clinical and follow-up data from 128 patients (142 eyes) with Eales disease undergone vitrectomy were retrospec tively analyzed. The statistical methods including t test,chi;2test, one-way Anova method of square-deviation(SD), and logistic regression were used to analyze the relationship between the general data of the patients (including age, sex, laterality of the eye, visual acuity before the treatment, stages of disease, duration from vitreous hemorrhage to vitrectomy, neovascularization and proliferative vitreoretinopathy, and whether or not combined with retinal detachment and other complications) and the prognosis of the visual acuity after surgery (including the surgical method, techniques, and times and complications after the surgery). The patients were 18-45 years old (mean 28.5 years) with single vitreous hemorrhage in 28 and proliferative changes in 114 in whom 59 had retinal detachment. The follow-up period after the surgery was more than 3 months (mean 35.8 months). The success criteria of the surgery were complete or part retinal reattachment, and failure of retinal reattachment, eye-ball atrophy or excis ion of the affected eye were the failure criteria. Results Successful vitrectomies had been performed on 129 eyes (90.8%) and unsuccessful ones on 13 eyes (9.2%). The difference between the stages of the disease and prognosis of visual acuity after the surgery was significant (chi;2=64.0, Plt;0.01); the duration of vitreous hemorrhage obviously affected the prognosis of visual acuity (OR=11.6，Plt;0.01); the degree, quality, curable possibility, and recurrent probability of combined retinal detachment were the key factors affecting the visual acuity after vitrectomy; the visual acuity before and after the surgery was interrelated; the method and techniques of the surgery and the different filling matters affected the visual acuity after the surgery; the difference between multiple times and once of surgery was significant (chi;2=66.84，Plt;0.01); the degree of complexity of the operative procedure, especially repetitious vitrectomies obviously affected the surgical prognosis and the improvement of visual acuity; the possibility of fa ilure of the surgery differs 7 times in patients with or without post-operative complications ( chi;2=67.23，Plt;0.01); whether the post-operative complications occurred or not significantly affected the prognosis of the visual acuity a-fter the surgery. Conclusions Vitrectomy is effective for Eales disease. The important factors affecting the prognosis of visual acuity after the operation include stages of disease, degree and extent of proliferative vitreoretinopathy, whether or not combined with retinal detachment and other complic ations, duration from vitreous hemorrhage to vitrectomy, the degree of complexity of the operation, and the complications during or after the operation. (Chin J Ocul Fundus Dis, 2006, 22:291-294)
Objective To evaluate the anatomical and functional results of retinotomy in treatment of complicated retinal detachment. Methods Twenty-three eyes were treated with retinectomy during vitrectomy for complicated retinal detachment with proliferative vitreoretinopathy when complete reattachment of retina was not obtained despite careful mambrane peeling. After released the peripheral vitreoretinal contraction or pulled subretinal membranes, perfluorocarbon liquid injection, laser retinopexy and silicone oil tamponade were performed. Retinotomy size ranged from 30-degree to 360-degree (average 132-degree). Results Retinal attachment were obtained in all of the operated eyes at the end of the operation. Silicone oil was removed from 15 eyes (65．0%) within 3～11 months postoperatively. After a minimum follow-up of 6 months, the retinae were completely attached in 17 eyes ( silicone oil was not removed in 4 eyes ) and visual acuity of 0.02 or better was obtained in 11 eye (48．0%). Recurrent retinal detachment occurred in 2 eyes in which the silicone oil had been removed. The major complications were recurrence of the proliferation in 6 eyes (26．0%) and hypotony in 3 eyes (13．0%). Conclusion Retinotomy is beneficial to reattach the retina for eyes with advanced proliferative vitreoretinopathy in seemin gly inoperable cases. (Chin J Ocul Fundus Dis,2001,17:87-89)