ABSTRACT
AIM: To observe and compare the statistical significance of superficial and deep vascular leakage in the pathological changes of the diabetic rats retina after the Evans blue (EB) perfusion, and utilize the modified whole-retina spreading method to make the slides while protecting the periphery of the retina. METHODS: The Sprague-Dawley (SD) rats were randomly divided into 6 groups. Each group named as the normal groups for 4, 8, and 12wk and the diabetic groups for 4, 8, and 12wk. The EB was injected into the cardiovascular system of the rats at the different time points. The retina of each group was obtained for observation. RESULTS: The superficial vascular leakage was found in all 6 groups. The size of leakage area of superficial retinal blood vessels was (0.54±0.23)%, (0.65±0.11)%, and (0.58±0.10)% in normal group. No notable leakage was found in the deep blood vessels [(0.03±0.04)%, (0.03±0.05)%, and (0.03±0.05)%]. The deep retinal vascular leakage was found in the peripheral retina of diabetic rats. The size of leakage area of superficial retinal blood vessels in diabetic group were (0.53±0.22)%, (0.69±0.16)%, and (0.52±0.11)%. The leakage areas of deep blood vessels were (0.54±0.50)%, (1.42±0.16)%, and (1.80±0.07)% at 4, 8, and 12wk, respectively. There was a statistically difference of the leakage area between the 8th week and the 4th week of diabetes group (P=0.003). The statistically significant difference between the diabetes and the control groups was noted at 4wk and 8wk (P<0.001). CONCLUSION: The main retinal pathological changes of early-stage diabetic rats are the vascular leakage of the periphery of deep retina. Diabetic rats modeled after 8wk have semi-quantitative statistical difference compared with the normal rats, thus early intervention treatment research can start at this time point.
ABSTRACT
AIM: To evaluate the long-term effects of pattern scan laser (PASCAL) pan-retinal photocoagulation (PRP) on diabetic retinopathy (DR) in Chinese patients. METHODS: In this retrospective study, we evaluated clinical data of 29 patients (53 eyes) with severe non-proliferative DR (SNPDR) or proliferative DR (PDR) who received PRP and follow-up at our hospital from 2008 to 2013. Sixteen patients (29 eyes) received PASCAL PRP and 13 patients (24 eyes) received 100-ms conventional laser PRP. RESULTS: After long-term follow-up (mean, min-max days: 719.8, 290-1666 for PASCAL PRP vs 743.5, 240-1348 for conventional PRP, P=0.569), patients receiving PASCAL PRP required fewer photocoagulation sessions than the conventional PRP group (2.6±1.0 vs 3.9±0.9, P<0.01). Best corrected visual acuity (BCVA) was reduced slightly in PASCAL PRP group while reduced significantly in conventional PRP group. At last visit, 24 eyes in the PASCAL group (88.9%) and 21 eyes in the conventional group (91.7%) were improved or stable. Two eyes in PASCAL PRP group (7.4%) and 3 eyes in the conventional PRP group (12.5%) developed vitreous hemorrhage or vitreous fibrovascular proliferation. CONCLUSION: PASCAL PRP is as effective and may be more conducive to maintaining visual acuity with less treatment sessions for DR treatment compared to conventional laser PRP.
ABSTRACT
To study insertion efficiency of goldfish Tgf2 transposon in the genome of Megalobrama amblycephala, we built Tgf2-Mlyz2-RFP donor plasmid with goldfish Tgf2 transposon left and right arms, and then co-injected with goldfish Tgf2 transposase mRNA into the 1-2 cell stage fertilized eggs of M. amblycephala. In 30 d- and 180 d-stage larval, RFP fluorescence can be observed in back and side muscle of the fish. The rate of RFP fluorescence expression was 48.1%. In adult fish, PCR results demonstrated that integration efficiency of goldfish Tgf2 transposition system was 31.5% in M. am-blycephala genome. RT-PCR analysis showed that RFP RNAs were highly transcribed among all the 12 tissues in three transgenic fishes, while it could be highly detected only in muscle, skin, and kidney in another two individuals. Our results suggested that RFP expression in tissues vaied among different M. amblycephala. By means of the inverse PCR, the copy numbers of Tgf2 transposon were at least 2 in transgenic M. amblycephala. The average copy number of each fish was about 5. Over 50% of flanking sequences at the insertion site have homologous sequence in other vertebrate species. Our data suggest that goldfish Tgf2 transposon can efficiently mediate gene insertion in M. amblycephala, which could been used in transgene and gene trap in M. amblycephala.
Subject(s)
DNA Transposable Elements , Transposases , Amino Acid Sequence , Animals , Base Sequence , Goldfish/genetics , Humans , Molecular Sequence Data , Mutagenesis, InsertionalABSTRACT
OBJECTIVE: To investigate the changes of vascular endothelial cell tight junction protein (occludin) and glial cell morphology as well as their relationship with blood-retinal barrier (BRB) in the retina of diabetic rats. METHODS: The distribution of occludin and GFAP were explored by immunofluorescence histochemical studies in the retina of streptozotocin (STZ)-diabetic rats and age-matched control rats. Evans blue was used to evaluate the impairment of BRB. RESULTS: GFAP immunoreactivity was limited to ganglion cell layer and nerve fiber layer in the control retina. GFAP immunoreactivity was significantly increased in ganglion cell layer and nerve fiber layer in one month diabetic rats. GFAP positive Müller cells were increased in three months and six months diabetic rats. Occludin immunoreactivity progressively decreased in three months and six months diabetic rats but not in the one month diabetic rats. Evans blue injection showed a progressive impairment of BRB. CONCLUSIONS: Astrocytes activation in the early stage of diabetes plays an important role in the maintaining of the BRB function. But the activation of Müller cells in the later stage destroyed the BRB eventually. These changes are consistent with the concept that BRB changes caused by altered glial-endothelial cell interactions contributes to the occurrence of diabetic retinopathy.
