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

Objective: Our aim was to investigate whether the immune RNA (iRNA) can inhibit the endotoxin-induced uveitis (EIU). Methods: The EIU was induced by injecting endotoxin of E.Coli into rats hypodermically. iRNA was abstracted from the livers and spleens of the rabbits which had been injected by E.Coli six times. The iRNA was introduced intraperitoneally into the rats both at half an hour and 3 hours after the injection of endotoxin. The effect of iRNA was evaluated by protein determination and cell count in aqueous houmer(AH). Results: The appearance of EIU in the iRNA treated rats was delayed, about 3 hours later than the rats received only endotoxin injection. The AH protein decreased by 58.34% and cellular infiltration decreased by 27.62% in iRNA treated group at the 24 hours. Conclusion: The iRNA may partly prevent the development of EIU and significantly reduce the severity of EIU.

Acetylcholine ; metabolism ; physiology ; Animals ; Cholinergic Agonists ; pharmacology ; Male ; Muscle Contraction ; drug effects ; physiology ; Muscle Relaxation ; drug effects ; physiology ; Muscle, Smooth ; drug effects ; pathology ; physiopathology ; Rabbits ; Random Allocation ; Receptors, Cholinergic ; physiology ; Synaptic Transmission ; drug effects ; Urinary Bladder ; drug effects ; innervation ; physiopathology

Amino Acid Metabolism, Inborn Errors ; blood ; diagnosis ; Humans ; Infant, Newborn ; Male

The aim of this study was to explore the effect of small hairpin loop RNA (shRNA) silencing hypoxia-induced factor 1alpha (HIF-1alpha) gene on the expression of vascular endothelial growth factor (VEGF) and pigment epithelium derived factor (PEDF) in human retinal pigment epithelium (RPE) cells under hypoxic condition. Two target sites of HIF-1alpha mRNA were chosen and two kinds of shRNA were designed and synthesized against the target sites. Then the two kinds of shRNA were transfected into human RPE cells in vitro, respectively. These cells were cultured under hypoxic condition that was simulated by using 150 mumol/L CoCl(2). The mRNA expressions of HIF-1alpha, VEGF and PEDF were tested by semi-quantitative reverse transcription PCR (RT-PCR). The protein levels of HIF-1alpha, VEGF and PEDF were analyzed by Western blotting. After the two kinds of HIF-1alpha-specific shRNA were transfected into RPE cells respectively, the expression of HIF-1alpha mRNA and the levels of HIF-1alpha protein were decreased significantly in RPE cells under hypoxic condition. The expression of VEGF mRNA and the levels of protein significantly were also decreased. However, the levels of PEDF protein was significantly increased, but the expression of PEDF mRNA showed no significant changes. In conclusion, HIF-1alpha-specific shRNA can effectively silence the HIF-1alpha gene, and consequently down-regulate VEGF and up-regulate PEDF expression against hypoxia. These results reveal that HIF-1 is associated with posttranslational mechanism for down-regulating PEDF under hypoxia and provide an explanation for hypoxia-provoked increases in VEGF/PEDF ratios. These results also suggest that HIF-1 is one of the key cytokines to retinal neovascularization.

In order to explore the effect of high glucose concentration and high glucose concentration with hypoxia on the production of hypoxia-inducible factor-1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF), human RPE cells were cultured in 5.56 mmol/L glucose (control group), 5.56 mmol/L glucose with 150 micro mol/L CoCl2 (hypoxic group), 25 mmol/L glucose (high glucose group) and 25 mmol/L glucose with 150 micro mol/L CoCl2 (combination group). RT-PCR was used to detect the expression of HIF-1alpha and VEGF mRNAs. Western blot analysis was used to measure the levels of HIF-1alpha and VEGF proteins. Although the small amount of HIF-1alpha protein was able to be detected in high glucose group but not in control group, there was no significant difference between the expression of HIF-1alpha mRNA of RPE cells in high glucose group and that of RPE cells in control group. As compared with RPE cells in control group, the mRNA expression and the protein synthesis of VEGF in high glucose group were up-regulated. As compared with RPE cells in hypoxic group, the expression of HIF-1alpha mRNA of RPE cells in combination group was not different, but the protein synthesis of HIF-1alpha, the mRNA expression and the protein synthesis of VEGF were more obviously up-regulated. In conclusion, high concentration glucose mainly influence the protein synthesis of HIF-1alpha of RPE cell, and HIF-1alpha protein is able to be accumulated in high concentration glucose. Under hypoxia, the HIF-1alpha protein induced by high concentration glucose is more stable, and the expression of VEGF is obviously increased. It is suggested that high concentration glucose may play a role in retinal neovascularization, especially at ischemia stage of diabetic retinopathy.

Objective To observe the transformation features of children with complete atrioventricular block(CAVB), and to improve the diagnosis and treatment of this disease.Methods Seventeen children with primary diagnosis of CAVB were reviewed by retrospective and follow-up study and the clinical characteristics, treatment schemes and prognosis were evaluated.Results Four cases of congenital CAVB lived normally without obvious symptoms in the tracing period from 6 months to 9 years;1 case had the onset of Adams-Stokes syndrome induced by diarrhea;3 cases of the CAVB caused by virus myocarditis turned into sinus rhythm after comprehensive therapy;2 cases persistently presented with the CAVB Complicated with enlarged heart;1 case gave up treatment after deterioration, and one case died.Three cases of temporary CAVB after the open heart operation turned to sinus rhythm in transitory time, while other two cases presented with permanent CAVB and treated with epicardium pacemaker,among whom one had the pacemaker replaced for one time in the 8-year follow-up,and the follow-up of other cases were intermitted.Conclusions The congenital CAVB in the study group with normal QRS interphase and no obvious symptom might not require treatment but follow-up is needed. While infants with heart malformation and wide QRS wave could not endure the low ventricular rhythm are in high risk. Virus myocarditis induced CAVB children tend to present the Adams-Stokes syndrome, and require effective treatments. Partial cases of the CAVB caused by open heart operation may turn to normal sinus rhythm in 2-4 weeks after surgery. cases has persistent CAVB for over 4 weeks, or Adams-Stokes syndrome onset after the surgery demand epicardium pacemaker treatment.

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) doubleenzyme-histochemistry staining to detect corneal lymphangiogenesis and hemangiogenesis. By immunohistochemistry for LYVE-1, it was found that blown lymphatics were localized in the stroma 3days 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 huma n 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.

China ; Dust ; analysis ; Female ; Humans ; Male ; Manufactured Materials ; Occupational Exposure ; prevention & control ; statistics & numerical data ; Occupational Health

PKS gene was screened by PCR from thirty strains of spone-associated bacteria including twenty-one actinomycetes isolated from Craniella anstrialiensis and nine bacillus isolated from Dysidea avara in the South China Sea.As a result,a 669 bp KS domain gene was successfully amplified from Bacillus C89.BLAST analysis showed that the KS domains were most closely related to the KS sequences of Bacillus subtilis subsp.subtilis str.168 with 96% similarity.Phylogenetic analysis demonstrated the KS domain belong to trans-AT KS domains.This study demonstrated the existence of PKS gene in bacteria associated with sponge Dysidea avara for the first time,and provided proof for the hypothesis that sponge-associated bacteria are perhaps the true producers of many novel bioactive compounds in sponge.Meanwhile,this study lays a basis for the microbial screening for polyketide compounds production.