Objective To explore the proper dosage of establishment of stable hepatic oval cells （HOC） prolif-eration model by using 2-acetaminofluorene （2-AAF） combined with two-third partial hepatectomy （2/3 PH） surgery， and to explore isolated and cultured method of HOC in vitro. Methods The 174 Wistar rats were randomly divided into 4 experimental groups （each group enrolled 30 rats）， saline group （n=30）， and untreated group （n=24）. Rats of 4 experi-mental groups were underwent gavage of 5， 10， 15， and 20 mg/（kg ? d） 2-AAF， corresponding to the groups from No.1 to No.4 group. Rats of saline group received saline gavage and rats of untreated group didn’t received any treatment. A standard 2/3 PH surgery was performed on the 5th day after gavage， then the same gavage method was still administrated as preoperation untill rats were sacrificed. The liver tissues of 6 selected rats were adopted and identified by HE staining and immunohistochemical staining on 4， 8， 12， and 16 days after PH for observation of the proliferation of HOC in every group， on 4 days， levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） were tested in addition. HOC were isolated and purified by collagenase perfusion method and percoll gradient centrifugation. Results The surv-ival rates of untreated group，saline group，No.1 group，No.2 group，No.3 group，and No.4 group were 100% （24/24），93% （28/30），93% （28/30），90% （27/30），90% （27/30），and 80% （24/30） respectively. Compared with the saline group and untreated group， the levels of serum ALT and AST increased significantly in No.2， No.3， and No.4 group on the 4th day after PH （P＜0.05）. The results of HE staining showed that No.2， No.3， and No.4 group were observed visibly different level of damage at liver tissue， and the proliferation level of HOC were most obviously in No.3 and No.4 group. The results of immunohistochemical staining revealed that proliferation cells were positively expressed oval cell marker-6 （OV-6）. The number of OV-6 positive cells were increased significantly with the increase of dosage of 2-AAF between 4 days and 12 days after operation， and proliferation levels were related with dosages of 2-AAF （P＜0.05）. In all cultured cells， 80% of cells were OV-6 positive cells after isolation and culture by using collagenase perfusion method and percoll gradient centrifugation. Conclusions The methods of gavage of 2-AAF at 15 mg/（kg ? d） combined with 2/3 PH surgery can establish the HOC proliferation model on the 12th day， as well as the rats have lower mortality and better tolerance， especially. The collagenase perfusion method and percoll gradient centrifugation can be used to isolate HOC effectively.
Objectives To investigate the expression of pax-6 in ret ina of in fant monkeys with myopia induced by optical defocus, and to determine the role of pax-6 would play or not in onset and development of myopia and emmetropization.Methods Nine healthy infant rhesus monkeys, aged from 1 to 3 months, were selected and wore spectacle lenses or underwent photorefractive keratectomy (PRK).Transcription polymerase chain reaction method and quantitative analysis were used to determine the expression of pax-6 in the retina with myopia induced by optical defocus in different time, and the result was compared with that in retina without myopia.Results The myopia caused by hyperopic defocus was found. The expression of pax-6 in the retina with myopia induced by optical defocus was significantly higher than that in the retina without myopia（t=3.480，P=0.004）.Conclusions The expression of pax-6 is enhanced by hyperopic defocus in the infant monkey retina, which suggests that pax-6 may be involved in vision-dependent eye growth and emmetropization. (Chin J Ocul Fundus Dis，2003，19：201-268)
Objective To explore the key technique of allogeneic whole pancreaticoduodenal transplantation (WPDT) in rats. MethodsWPDT model was established between Lewis rats as donors and Wistar rats (with type 1 diabetes mellitus) as recipients. End to side anastomosis was performed in abdominal aorta of donors and recipients. The portal vein of the graft was anastomosed with the recipients left renal vein by cuff technique. And side to side anastomosis was made between the graft duodenum and the host jejunum. ResultsForty-four of 50 rats were successfully performed WPDT. Amongthem, 8 rats died in postoperative 3 days, the survival time of residual 36 rats was 6-16 days, with an average of (10.45±3.30) days. The peak of death appeared on day 7-10 after operation. The typical acute rejection in pathological changes were observed on day 7. ConclusionSkilled microsurgical techniques and emphasis on details are important to establish WPDT model.
Objective To investigate the expression of T cell receptor (TCR) Vβ8.3 gene on CD4+ T lymphocytes in the rats with experimental autoimmune uveoretinitis (EAU). Methods Eighteen Lewis rats were divided into EAU, complete Freund′s adjuvant, and the control group. Inter photoreceptor retinoid-binding protein (IRBP) R16 peptide was synthesized using Fmoc procedure for induction of EAU. Magnetic absorption cell sorting (MACS) me thod was used to isolate the CD4+T lymphocytes from the spleen of the rats. Flow cytometry was used to monitor the efficiency of isolation. The expression of TCR Vβ8.3 gene segment on CD4+T lymphocytes was determined by fluorescent quantitative polymerase chain reaction. Results EAU was successfully induced in the Lewis rats immunized with IRBP R16 peptide. The proportion of CD4+T lymphocytes isolated by means of MACS was statistically higher than that before isolation (P＜0.001). The expression of TCR Vβ8.3 gene segment on CD4+ T lymphocytes in EAU rats was significantly higher than that in the control (P＜0.05). Conclusions There is a predominant usage of antigen-specific TCR Vβ 8.3 gene in EAU rats induced by IR BP R16 peptide, which may serve as a target for immunotherapy of EAU. (Chin J Ocul Fundus Dis，2004，20：165-167)
A acute partial obstructive hepatocholangitis model by selective ligation and injection of E coli into left hepatic bile duct was successfully founded in rat. Using parameters including mortality, mitochondrial glutamic oxalacetic transaminase and ornithine carbamoytransferase activity, pathological observation and blood culture of bacteria, we evaluated the model. The authors emphasize that this models is superior to the wole-bile-duct-challenged cholangitis model, which is characterized by liver injury.
Purpose To determine the effect of exogenous interleukin-1alpha; (IL-1alpha;) on the retina and its vasculature and VEGF expression in SD rats． Methods IL-1alpha;2．0 ng (20 mu;l) were injected into the vitreous of 8 left eyes of 8 SD rats while steriled PBS were injected into 8 right contralateral eyes of the same rats as control． All eyes were assessed by direct ophthalmoscopy every day and enucleated on the 7 thpostoperative day． Histological examination (hemato xylineosin staining) and immunohistochemical staining with antibody against VEGF antigen were performed， and sections were observed and photographed under light microscopy． Results ①All 8 IL-1alpha; inject ed eyes developed epiretinal membranes and extraretinal neovascularization on the 3 rd postoperative days while none of the 8 control eyes exhibited any a bnormal retinal vascular changes and they were confirmed by HE staining;②Immuno staining identified VEGF express mainly in the inner layer of vessel walls， the epiretinal membranes， the neuroganglional layer and the photoreceptor layer of retina， while the control eyes showed only weak positive staining in the photo receptor layer． Conclusions IL-1alpha; is capable of inducing vitreo retinal neovascularization，and increasing the expression of VEGF in the retina and epiretinal membranes． (Chin J Ocul Fundus Dis, 2001,17:135-137)
【Abstract】Objective To investigate the effect of ischemia on the treatment of advanced body and tail carcinoma of pancreas. Methods In operation the proximal spleen artery was ligated, a chemotherapy pump was installed to connect the distal spleenic artery and urea solution (40%) was injected through the device during and after operation. The pathology and pathophysiology change in dogs were observed. Results All the eight dogs studied were alive after operation, no serious complication appeared, pancreatic cells were replaced by fibrosis. Conclusion Infusion of 40% urea solution is a safe and effective ischemic method, it can lead to all sorts of pancreatic cell necrosis and fibrosis. It may be a good madality in the treatment of advanced pancreatic carcinoma.
Objective To investigate the effect of inducible nitric oxide synth ase (iNOS) or cyclooxygenase－2 (COX－2) on retinal neovascularization and its possible mechanism in oxygen－induced retinopathy （OIR） mouse model. Methods Retinal neovascularization was induced by oxygen with different concentration. The expression of iNOS, COX－2, matrix metalloproteinases 2 (MMP－2) and vascular end othelial growth factor (VEGF) in the retinae of experimental animals were analyzed by immunohistochemistry, realtime polymerase chain reaction and western blotting technologies. Results The inhibition of COX－2 or iNOS obviously attenuated retinal neovascularization and decreased the expression of VEGF and MMP－2. The iNOS inhibition decreased COX－2 expression, and vice versa. Conclusions COX－2 and iNOS may play a role in retinal neovascularization in OIR mouse model, which may act by regulating the expression of VEGF and MMP－2.