In order to investigate the ipsilateral lymphadenectomy for inhibiting rejection in rat corneal transplantation, corneal allogenic transplantation models were established in rats. Eighteen female Wister rats were used as donors, and 36 Sprague Dawley rats as recipients. After penetrating corneal transplantation, recipients were randomly divided into 3 groups: group A (control group); group B, the ipsilateral lymphadenectomy group; group C, the bilateral lymphadenectomy group. Among 12 rats in each group, the corneas of 2 rats in each group were used for pathological study at day 14 after the transplantation, and the remaining 10 rats were used for studying corneal rejection by a slit lamp. The time points when allograft rejection occurred were recorded and mean survival time (MST) was compared. The results showed that MST in groups B and C was 46.30 +/- 9. 464 days and 44.43 +/- 7. 604 days, respectively, which was significantly prolonged as compared with that in group A (10.71 +/- 1.567 days, P < 0.01). There was no significant difference in MST between groups B and C (P > 0.05). It was concluded that both bilateral and ipsilateral lymphadenectomy therapies could effectively inhibit the corneal allograft rejection. Ipsilateral lymphadenectomy is a less complex surgical procedure and is just as effective in preventing rejection.
Corneal Transplantation ; Graft Rejection/*prevention & control ; Graft Survival ; Lymph Node Excision/*methods ; Random Allocation ; Rats, Sprague-Dawley ; Rats, Wistar

In order to investigate the effects of mouse CTLA4Ig gene-modified dendritic cells (DCs) on the survival of the corneal allografts in rats, the plasmid PG\CTLA4Ig was transfected into DCs of F344 rats mediated by Lipofectamine 2000. The expression of CTLA4Ig was detected by immunofluorescent microscopy. The effects of donor DCs on the proliferation of T cells in Lewis rats (recipients) were tested by by CCK8. Corneal transplantation was performed from F344 rats to Lewis rats. The DCs modified with CTLA4Ig gene were injected into the Lewis rats on the day 0 and 3 after transplantation. The movement of the DCs after modification in vivo was observed by immunofluorescent microscopy, and the survival of corneal allografts was evaluated by Holland criterion. The results showed that the CTLA4Ig-modified DCs could restrain the proliferation of allogenetic T cells. The CTLA4Ig-modified DCs prolonged survival of corneal allografts. (P < 0.01). It was suggested that the injection of CTLA4Ig gene-modified DCs could obviously inhabit the allograft rejection and prolong the survival of corneal allografts.

Objective To investigate the efficacy of ganciclovir in viral keratitis and its influence on serum related indicators.Methods Patients with viral keratitis from May 2012 to December 2013 in our hospital were randomly divided into control and observation groups, each group had 62 cases.Patients in control group were treated with acyclovir eye drops, and those in observation group were treated with ganci-clovir ophthalmic gel.The difference in efficacy of two groups was compared.The levels of serum inflamma-tory cytokines of interleukin-2 (IL-2), IL-4, IL-6, IL-8, tumor necrosis factor-α(TNF-α), and interferon-γ( IFN-γ) of two groups before and after treatment were compared, the serum levels of nitric oxide ( NO) , super oxide dismutase (SOD), malondialdehyde (MDA), C3, and C-reactive protein (CRP) were been compared.Results The pain relief time and corneal healing time of observation group [(3.41 ±0.89)d and (4.02 ±1.11)d] were all shorter than control group [(4.52 ±1.21)d and (6.30 ±1.45)d].The cure rate and total effective rate of observation group were 58.06%and 90.32%, respectively.It was sig-nificantly higher than control group ( P <0.05).After 3 and 7 days of treatment, the serum levels of IL-4, IL-6, and IL-8 were lower than control group, and IL-2, and IFN-γwere higher than control group (33.87%and 82.26%) ( P <0.05).After 3 and 7 days of treatment, the serum levels of NO and MDA were lower than control group, and SOD, C3, and CRP were higher than control group ( P <0.05).Con-clusions Ganciclovir has a better efficacy than acyclovir for viral keratitis, and can effectively adjust the serum levels of related indicators.

AIM: To explore the effect of protein kinase C-?(PKC-?) in U937 cell line inhibited by short hairpin RNA(shRNA) on the transcription of vascular endothelial growth factor(VEGF) mRNA induced by stromal cell-derived factor-1(SDF-1).METHODS: 64 bp reverse repeated motifs of PKC-? target sequence were synthesized and inserted into the plasmid to construct the plasmid expressing shRNA-PKC-?(pSIRENp) and the pSIRENp plasmid was transfected into U937 cell line.The expression of PKC-? and VEGF mRNA was detected by RT-PCR.RESULTS: The recombinant plasmid pSIRENp was successfully constructed and it nearly completely suppressed the PKC-? expression in U937 cell line.After transfection,both basical and VEGF mRNA induced by SDF-1 significantly reduced compared to control.CONCLUSION: The results shows that the short hairpin RNA of PKC? efficiently reduces its expression in U937 cells and PKC-? may be involved in the regulation of VEGF mRNA expression.

Based on Career-Life-Cycle theory and practical experiences,this paper analyzed characters and varied requirements in scientific research during doctors' career development,and discussed the way and effects to construct talent cultivation mode of Research-Life-Cycle for doctors in hospitals.

In order to construct a recombinant plasmid containing short hairpin RNA (shRNA) targeting survivin and to investigate its effect on survivin expression and cell apoptosis of human retinoblastoma cell line Hxo-rb44 in vitro, RNA interference plasmid pSIRENS that can express shRNA of survivin was designed, constructed, and transfected into human retinoblastoma cell line Hxo-rb44. Survivin and c-Myc expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Apoptosis of Hxo-rb44 cells was assayed by Honchest33258 staining and cell growth curve was drawn. The results showed that the oligonucleotide targeting survivin was identified in pSIRENS plasmid. After pSIRENS plasmid transfected, survivin and c-Myc expression in Hxo-rb44 cells was decreased significantly. Apoptotic rate of cells was up-regulated from (3.5+/-1.29) % to (36.1+/-19.66) %. The proliferation ability of Hxo-rb44 cells was inhibited. No significant effects on survivin expression and apoptosis of the cells were found when negative control plasmid was transfected. In conclusion, the plasmid containing shRNA targeting survivin was constructed successfully. It could inhibit efficiently the expression of survivin and c-Myc in human retinoblastoma cell Hxo-rb44 in vitro. The inhibition of the expression of c-Myc might be involved in the apoptosis of Hxo-rb44 cells.

To evaluate the efficacy and safety of resection and cryotherapy combined with amniotic membrane transplantation (AMT) for the treatment of vernal keratoconjunctivitis (VKC) with giant papillae (GP). Eight patients (16 eyes involved) with VKC, characterized by GP on the upper tarsal conjunctiva, underwent resection and cryotherapy in combination with AMT. The follow-up lasted for 3-22 months. The results showed that corneal shield ulcers and superficial punctuate keratitis healed during the first week after surgery and did not recur. Fourteen eyes (87.5 %) were symptom-free 1 month after surgery, and no GP, ectropion, trichiasis and other complications were noted, but the blood vessels of upper tarsal conjunctiva could not be clearly seen and a little conjunctival scar was observed. Recurrence of GP was observed in 2 eyes (12.5 %), with the area being less and irritation milder as compared with those before the operation. Among the two eyes, one eye was treated by cyclosporine eyedrops with improvement, but the other eye showed no improvement after the treatment, and underwent a second surgery with a cotton patch soaked in fluorouracil applied onto the supratarsal area after resection and cryotherapy. Four months after the treatment the patient presented no symptoms and GP did not recur. It is concluded that the resection and cryotherapy combined with AMT is an effective and safe treatment for VKC with GP.

To study corneal lymphangiogenesis after corneal transplantation, corneal allogenic transplantation models were established in rats. 8 female Wister rats were used as donors, and 16 Sprague Dawley (SD) rats were used as recipients and 2 SD served as controls. Corneal lymphangiogenesis and hemangiogenesis was examined by electron microscopy 1 and 2 weeks after corneal penetrating transplantation, and the expression of lymphatic vessel endothelial receptor (LYVE-1) was examined 1, 3, 7, 14 days after the transplantation respectively. In addition, 19 allograft failed human corneas were examined by 5'-nase-alkaline phosphatase (5'-NA-ALP) double-enzyme-histochemistry staining to detect corneal lymphangiogenesis and hemangiogenesis. By immunohistochemistry for LYVE-1, it was found that blown lymphatics were localized in the stroma 3 days after the corneal transplantation. With electron microscopy, new lymphatic vessels and blood vessels were found 1 and 2 weeks after the corneal transplantation. By 5'-NA-ALP enzyme-histochemistry, corneal hemangiogenesis was found in all allograft failed human corneas and 5 of 19 (26.3 %) cases had developed corneal lymphangiogenesis. It is concluded that corneal lymphangiogenesis is present after corneal transplantation, which may play an important role in allograft rejection.
Cornea/*blood supply ; Cornea/chemistry ; Cornea/ultrastructure ; Corneal Neovascularization/etiology ; Corneal Neovascularization/metabolism ; Corneal Transplantation/adverse effects ; Corneal Transplantation/*methods ; Immunohistochemistry ; Lymphangiogenesis ; Microscopy, Electron ; Rats, Sprague-Dawley ; Rats, Wistar ; Vesicular Transport Proteins/biosynthesis

Smooth muscle cell (SMC) proliferation, platelet free calcium level and aggregation of experimental atherosclerotic rabbits were investigated in this study. Aortic SMC ofhyperlipidemic rabbits in vitro showed higher growth activity than did normal rabbit SMC. And also hyperlipidemic serum stimulated SMC to proliferate at a significantly greater rate than control serum. Moreover, the level of platelet free calcium and the platelet aggregation was also higher in hyperlipidemic rabbits, indicating that activitated platelets possibly release more PDGF to act as a stimulator to SMC proliferation and calcium is an important factor to activate platelets. Furthermore, SMC from 8501-treated rabbits appeared lower proliferative rate than thecells from hyperlipidemic rabbits. And serum from those rabbits inhibited SMC proliferation compared with hyperlipidemic serum, the inhibitory effect was even stronger than that of normal serum. It may be relevant to the favorable effects of 8501 to TXA2/PGI2 balance.

It has proved to be difficult for administrators of scientific research to improve process management,especially to manage cooperative research projects accomplished by various institutions.In this paper,we took the process management of Scientific Research in Public Interest (SRI-PI) based on a Grade 3A hospital and other 19 institutions as an example,to analyze the application of PDCA cycle in projects management.It is concluded that the comprehensive quality management of research projects should follow the scientific procedure of PDCA cycle.