Based on the requirements of Good Preparation Practice in Medical Institutions(GPP) and considering the actual conditions of the hospital,we decided upon a right way in the reconstruction of the hospital's preparation room and amelioration of the equipment and apparatus that constituted 30% of the investment.The results were reduced investment,higher efficiency,with perfect conformity to the GPP requirements.

Objective To investigate whether MEG3 involved in the development of retinoblastoma by down-regulating the expression of P53 protein.Methods The MEG3 expression of retinoblastoma tissues and corresponding non-tumor tissues were detected by quantitative real-time PCR (qRT-PCR).Retinoblastoma cell lines SO-RB50 or HXO-RB44 were transfected with pcDNA-MEG3 or siRNA-MEG3,after which cell apoptosis was tested by flow cytometry and P53 protein expression was tested by Western blot.Results MEG3 expression of retinoblastoma tissues was significantly reduced compared with corresponding non-tumor tissues(P =0.014).MEG3 level was significantly increased in pcDNA-MEG3 transfected SO-RB50 cells (P =0.002) and significantly decreased in siRNA-MEG3 transfected HXO-RB44 cells (P =0.004).Flow cytometry showed that the SO-RB50 cells apoptosis was significantly increased with the MEG3 over-expression(P ＜ 0.05),as well as the HXO-RB44 cells apoptosis was significantly decreased with the MEG3 knockdown(P ＜ 0.05),compared with the control group,respectively.Furthermore,Western blot showed that P53 protein level was significantly increased after SO-RB50 transfected with pcDNA-MEG3 (P ＜ 0.05),while significantly decreased after HXO-RB44 transfected with siRNA-MEG3 (P ＜ 0.05),compared with the control group,respectively.Conclusion MEG3 is down-regulated in retinoblastoma,affect the development of retinoblastoma,and may induce the retinoblastoma cell apoptosis by promoting the expression of P53 protein.

BACKGROUND:Corneal neovascularization not only seriously affects vision, it is also a high-risk factor for the rejection after allogeneic ceratoplasty, thus it is always a hot issue and in ophthalmology to investigate the pathogenesis of corneal neovascularization and the inhibitors for blocking its formation.OBJECTIVE: To induce model of corneal neovascularization in rats using nylon suture, and investigate the mechanism of nuclear factor-кB (NF-кB) in the occurrence and development of corneal neovascularization using dexamethasone as the glucocorticoid inhibitor for corneal neovascularization.DESIGN: A randomized controlled trial.SETTING: Department of Ophthalmology, Zhujiang Hospital of Southern Medical University.MATERIALS: The experiments were carried out in the Zhujiang Hospital of Southern Medical University from January to April 2005. Fifty-five healthy male Wistar rats of clean degree were used. Rabbit-anti-rat NF-кB P65 monoclonal antibody, rabbit-anti-rat vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-1)monoclonal antibodies, and 3,3-diaminobenzidine (DAB) kit were purchased from Wuhan Boster Biological Technology Co., Ltd.METHODS: ① Interventions: The rats were randomly divided in the saline control group (n =25), dexamethasone group(n =25) and normal comea group (n =5). Corneal neovascularization using nylon suture was induced in rats in the saline control group and dexamethasone group, and then saline and dexamethasone was dropped to the right eye of the rats respectively, 2 drops for each time, 3 times a day for 18 days. Not any treatment was given to the rats in the normal cornea group. ② Evaluations: The score of corneal neovascularization was evaluated in the saline control group and dexamethasone group at 1, 3, 7, 12 and 18 days postoperatively. Corneal sections were prepared to observe the histological changes of cornea under light microscope; The expressions of NF-кB, VEGF and ICAM-1 were detected with immunohistochemical staining.MAIN OUTCOME MEASURES: ① Score of corneal neovascularization; ② Histological changes of cornea; ③ Expressions of NF-кB, VEGF and ICAM-1 in cornea.RESULTS: ① Score of corneal neovascularization: The corneal neovascularization was obviously inhibited, and scores of corneal neovascularization at different time points were all significantly lower than those in the saline control group (P ＜0.05-0.01). ② Histological changes of cornea: In the dexamethasone group, corneal neuvascularization and the infiltration of inflammatory cells after suture were obviously alleviated as compared with those in the control group, and the corneal structures in each layer were relatively complete. ③ Expressions of NF-кB, VEGF and ICAM-1 in cornea: In the dexamethasone group, the expressions of NF-кB, VEGF and ICAM-1 at each time points were all lower than those in the saline control group (P ＜ 0.05). The intensity of the expression of NF-кB was positively correlated with the score of corneal neovascularization and the expressions of ICAM-1 and VEGF (r =0.961, 0.922, 0.958, P ＜ 0.01).CONCLUSION: NF-кB is involved in the formation of corneal neovascularization possibly through upregulating the expressions of its downstream genes (VEGF, ICAM-1). Dexamethasone can inhibit the expressions of many factors related to corneal neovascularization regulated by NF-кB, including cytokines and adhesion molecules, through reducing the activity of NF-кB, and then suppresses the occurrence and development of corneal neovascularization.

Objective: To study the effect and mechanism of Rhodobryum giganteum (Schwaegr.) Par. 's anti-atherosclerotic effect. Methods:Vascular endothelial ceils were cultivated and the H2O2 were used to induce the oxidative stress injury of human umbilical vascular endothelial cells (HUVEC). Water extract of Rhodobryum giganteum (Schwaegr.) Par. Was added to cultivated HUVEC, and the activity of the cells was carefully determined by OD value with MTF method. The Griess Reagent was used to detect the NO concentration of different groups. At the same time, the NO fluorescence analysis probe, DAF-FM DA. (3-amino, 4-aminomethyl-2', 7'-difluorescein, diacetate), was used to determine the activity of NO synthase. Results:The most suitable stimulating concentration of H2O2 on HUVEC is 12.5 mmol· L-1. Water extract of Rhodobryum giganteum (Schwaegr.) Par. Was co-cultured with HUVEC damaged byH2O2 .The OD values indicate that 3.33,2.50 mg· mL-1 water extract groups increase activity of the cells significantly compared with the model group (P <0.05) ,and 3.33 mg·mL-1 is much better than 2.50 mg·mL-1 dose group. To determine the content of NO and NOS, the 2.50 mg·mL-1 dose has significant effect (P < 0.05). Conclusions: The H2O2 concentration of 12.5 mmol· L-1 could be used to establish the injured model of endothelial cell successfully. 2.50 and 3.33 mg·mL-1 water extract of Rhodobryum giganteum (Schwaegr.) Par. Have protective effect on ECV304 injured by H2O2. 2.50 mg· mL-1 water extract of Rhodobryurn giganteum (Schw aegr.) Par could increase the activity of NOS and promote the synthesis and secretion of NO.

BACKGROUND: Nuclear factor-kappa B (NF-κB) plays a key role in regulating expressions of cytokines and adhesion factors during transcription and in central adjustment during corneal graft rejection reaction.OBJECTIVE: To study the dynamic expressions of NF-κB, intercellular adhesion molecular 1 (ICAM-1) and vascular endothelial growth factor (VEGF) in corneal graft and to investigate the interventional effect of cyclosporin A (CsA). DESIGN, TIME AND SETTING: A randomized controlled animal study was performed in the Department of Ophthalmology, Zhujiang Hospital, the First Military Medical University of Chinese PLA between January and July 2005.MATERIALS: 40 SD rats and 50 Wistar rats were included and randomly divided into three groups, syngenic transplantation (10 Wistar rats used as donor and another 20 Wistar rats used as acceptor), allogeneic transplantation (10 Wistar rats used as donor, and 20 SD rats used as acceptor), and allogeneic transplantation+CsA treatment (10 Wistar rats used as donor, and 20 SD rats used as acceptor).METHODS: Comeal transplantation models were established. Gentamicin (2 000 U) was subconjunctivally injected into the experimental eyes of all ecceptors every other day for three times in total; chloroptic (2.5 g/L) was then used two droplets every time, twice a day, and 18 successive days in total; additionally, tropicamide (5 g/L) was also used two droplets every time, once a day, and 7 successive days in total. One day after corneal transplantation, CsA eye droplet (10 g/L) was used in the allogeneic transplantation+CsA treatment group two droplets every time, three times a day, and 18 successive days in total.MAIN OUTCOME MEASURES: Scores of corneal graft rejection reaction index were measured at day 3, 7, 12, and 18 after corneal transplantation; pathological changes of corneal graft were observed at the same time points to detect expressions of NF-κB, ICAM-1, and VEGF.RESULTS: Rejection reaction was not observed in the syngenic transplantation group at 18 days; however, indexes of rejection reaction in the allogeneic transplantation group were higher than those in the syngenic transplantation group at four time points (P<0.05), but indexes in the allogeneic transplantation+CsA treatment group were lower than those in the allogeneic transplantation group (P<0.05). Immunohistochemical examination indicated that NF-κB, ICAM-1, and VEGF were located at corneal epithelial lamina and substantia propria layer and in newborn vascular endothelial cells. At each time point, expressions of NF-κB, ICAM-1, and VEGF in the allogeneic transplantation group were higher than those in the syngenic transplantation group (P<0.05) but lower than those in the allogeneic transplantation+CsA treatment group (P<0.05).CONCLUSION: CsA can weaken nuclear translocation and activity of NF-κB to inhibit expressions of cytokines, adhesion molecules, and other relative factors so as to inhibit ccurrence and development of corneal graft rejection reaction.

BACKGROUND:Al ogeneic penetrating keratoplasty is the most effective method for treating corneal blindness. However, the incidence of rejections is high after keratoplasty, so it is urgent to develop an immunosuppressive drug with high efficacy and low toxicity. OBJECTIVE:To establish al ogeneic penetrating keratoplasty models and monitor the expression of tumor necrosis factor-αin blank control group and after transforming growth factor-β1 eyedrop during acute rejection period of corneal grafts. METHODS:Al ogeneic penetrating keratoplasty models were established and were randomly divided into blank control group, ciclosporin A group (1%ciclosporin A), and transforming growth factor-β1 group (1μg/ml transforming growth factor-β1 eyedrop). The medications from each group commenced at 1 day after surgery, one eyedrop once, three eyedrops per day. Al the operated eyes were given 0.3%ofloxacin ophthalmic solutions and 0.5%tropicaide ophthalmic solution, three times per day, for 12 days. The corneal grafts were harvested for hematoxylin-eosin staining and immunihistochemical staining (SABC method), to detect tumor necrosis factor-αexpression in corneal grafts. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that, corneal grafts were significantly thickened, a large number of histoleucocytes and lymphocytes infiltrated in the blank control group;corneal grafts showed normal thickness and no inflammatory cel s infiltrated in the transforming growth factor-β1 group. Immunohistochemical staining showed that, there were less cel s positive for tumor necrosis factor-αin the transforming growth factor-β1 group compared with the blank control group (P<0.05). Transforming growth factor-β1 eyedrops can reduce the expression of tumor necrosis factor-αin the corneal grafts during acute rejection period, and reduce the inflammatory cel s infiltration in the corneal grafts, which is probably the mechanism of transforming growth factor-β1 to prevent and treat corneal al ograft rejection.

BACKGROUND:The preparation of recombinant human epidermal growth factor (rhEGF) and basic fibroblast growth factor (bFGF) has been clinical y used in the repair of ocular surface trauma. However, the concentration of these growth factors that achieve the maximal healing effect and the comparison of two kinds of growth factors on promoting wound healing are stil controversial.
OBJECTIVE:To investigate the effect of rhEGF and bFGF on the cloning of human corneal epithelial cells.
METHODS:The human corneal epithelial cells cultured in vitro were interfered with rhEGF and bFGF under different concentrations. The proliferation of human corneal epithelial cells was detected using MTT assay after 3, 5, 7 days of growth factors treatment. Plate clone formation assay was applied to observe the morphology of cellclone and analyze clone formation rate of human corneal epithelial cells.
RESULTS and CONCLUSION:MTT value shows that 10μg/L rhEGF and 10μg/L bFGF on day 5 were the most powerful concentration. The clone formation rate of human corneal epithelial cells after treated with 10μg/L rhEGF was higher than that with 10μg/L bFGF (P=0.02). The results confirmed that both rhEGF and bFGF could promote the proliferation and increase clone formation ability of human corneal epithelial cells. 10μg/L rhEGF for 5 days achieves the best effect on promoting clone formation of human corneal epithelium cells.

BACKGROUND:Currently,there are many studies concerning the pathogenesis,process,and damage of glaucoma,however,there is not an ideal glaucoma modelOBJECTIVE:To prepare rabbit intraocular hypertension models using three different material injections,and to verify the practical value of intraocular hypetension modelsMETHODS:Thidy New Zealand rabbits were divided randomly into 3 groups,with 10 animals in each group.One eye of each rabbit was served as the experimental eyes and the other eye as control eyes.Autoblood.methyl cellulose,C3F8 was injected into the anterior chamber of the experimental eyes.and the normal saline was injected into the control eyes.The intraocular pressure(IOP)was monitored prior to injection and at hours 0,24,36,48,72,96.120 and 168 after injection.RESULTS AND CONCLUSION:Intraocular hype Rension models could be induced by injecting 3 kinds of materials,and the IOP was obviously increased after injection(P＜0.05),and the ranges and periods of increasing were varied.The periods of increasing of 3 materials were 1,3 and 7 days,respectively,which could maintain for longer time for a second injection.The IOP ranged 1.86-6.65 kPa,and mild anterior segment inflammation could be found.The experiment demonstrated that intraocular hypeansion models using three different material injections are ideal models,which is characterized by simple,reliable and controllable.The suitable model can be selected for acute or chronic glaucoma research.

Objective To analyse the clinical value of bronchial artery embolization in treatment of massive hemoptysis.Methods 87 patients with hemoptysis included bronchiectasis in 46 cases,pulmonary tuberculaosis in 18 cases,bronchial carcinoma in 15 cases,bronchial arteriovenous malformation in 2 cases and unknown hemoptysis in 6 cases underwent embolized treatment of bronchial arteriography or intercostal arteriography.Of them,bronchial artery embolization were performed in 78 cases,intercostal artery embolization were done in 6 cases,both embolization of bronchial artery and intercostal artery were in 3 cases,2 cases underwent superselective embolization with coaxial microcatheter.In the total 87 cases,gelatin sponge particles(GSP) alone was used in 85 cases,GSP and polyvinyl alcohol(PVA) were used in 2 cases.All the patients were followed up for 12~18 months.Results The hemoptysis was immediately stopped in 58 cases, remarkable improvement were 19 cases after operation. Recurrence of hemoptysis was found in five, three and two cases at one, two and four weeks respectively after embolization. All of the ten recurred cases accepted re-embolization and hemoptysis had been well controlled. The effective rate was 89%(77/87). There was not any severe complication in all the patients.Conclusion Bronchial artery embolization is a safe, effective and less invasive method in treatment of massive hemoptysis. It can be recommended to the non-surgical treatment patients.

Objective To establish an HPLC fingerprint of Herba Rhodobryi Rosei.Methods The fingerprint chromatography has been determined by RP-HPLC.The analysis was carried out with Dikma ODS C18 column(250 mm?4.6 mm,5 ?m).The mobile phase were:0.5% HAcCN(A),0.5% HAcH2O(B);Elution method:0-50 min,A was 20%-55%;50-60 min,A was 55%-95%;60-85 min,A was 95%-100%;keeping 5 min.Flow-rate was 1.0 mL/min.Wavelength was 360 nm and temperature was 30 ℃.It was analysized with the Estimating System of Similarity of 2004A Version(the Country's Pharmacopeia Committee)on the Chinese Medicine Fingerprint Chromatography.Results The fingerprint chromatography of Herba Rhodobryi Rosei was estabilished.Conclusion The method can be used in quality control of Herba Rhodobryi Rosei with accuracy and better repeatability.