ObjectiveTo investigate the expression of miR-195 and the underlying molecular mechanisms of miR-195 regulating HMGB1 in diabetic retinopathy (DR). MethodsExtract 5 ml venous blood from DR patients, diabetes mellitus (DM) patients and normal subjects, then extract and perificate plasma total RNA. MicroRNA array and real time polymerase chain reaction (RT-PCR) was used to screen out miRNAs which were expressed with significant differences in the serum of patients with DR. Bioinformatics was employed to predict the miR-195 related to high mobility group box 1 (HMGB1) regulation. Next, miR-195 was down-regulated or up-regulated in umbilical vein endothelial cells through transfection of miR-195 inhibitor and miR-29b mimics respectively.Then we analyzed expression of HMGB1 mRNA and protein by RT-PCR and Western blot. ResultsMicroRNA array results showed the expression of miR-195 in DR group is decreased by 8.34 times and 11.47 times compared with DM group and the normal group. RT-PCR verification results conforms to the microRNA array results. Compared with the DM group (F=0.034, t=8.057) and the normal group (F=0.370, t=9.522), the expression of miR-195 in DR group were significantly reduced, the differences were statistically significant (P < 0.05). RT-PCR showed that the expression of HMGB1 mRNA was significantly decreased in up-regulation group, compared with blank (F=0.023, t=11.287) and negative control group (F=0.365, t=7.471), the difference was statistically significant (P < 0.05). The expression of HMGB1 mRNA was significantly increased in down-regulation group, compared with blank (F=0.053, t=10.871) and negative control group (F=0.492, t=6.883), the difference was statistically significant (P < 0.05). Western blot showed that the expression of HMGB1 protein was significantly decreased in up-regulation group, compared with blank (F=0.021, t=8.820) and negative control group (F=0.039, t=7.401), the difference was statistically significant (P < 0.05); and significantly increased in down-regulation group, compared with blank (F=0.186, t=10.092) and negative control group (F=0.017, t=12.923), the difference was statistically significant (P < 0.05). ConclusionMiR-195 can inhibit the expression of HMGB1, reduce the inflammation and angiogenesis, thereby delaying or inhibiting the occurrence and development of DR.
Optogenetics is a novel technique which combines optics with genetics. Using genetic means, a selected opsin protein is ectopically expressed in target neurons, which are then stimulated by light to moderate the neuronal circuit, as a consequence to regulate the animal's behaviors. Retinal degeneration like retinitis pigmentosa and aged macular degeneration causes visual impairment and eventual blindness. Optogenetics techniques have opened the door to creating artificial photoreceptors in the remaining retinal circuits of retinal degeneration retinas via gene therapy. However, there are still limitations in optogenetics technique, for example, potential risk in virus infection, the choice of target cells and the low visual resolution of the experiment animal. It has been reported that vision was successfully restored to a certain extent in animal model using optogenetics technique. With higher photosensitivity of opsin protein, longer activation kinetics and higher transfection efficiency of virus vector, optogenetics techniques' application in ophthalmology will be improved.
Objective To investigate the feasibility of differentiation of invitro induced rat bone marrowderived mesenchymal stem cells（rMSCs） into retinal pigment epithelial （RPE） cells.Methods The rMSCs from BrwonNorway (BN) rats were isolated and cultured by adherent screening method. RPE cells lysate made by repeated freezethawing was put into the rMSCs culture system to identify whether the induced cells could express characteristic label cytokeratin（CK）and S-100 simultaneously or not.Results The growth rate of rMSCs induced by RPE cells lysate was slower and protuberant burr surrounded the fusiform cells. The results of immunoblotting and double immunofluorescence showed that partial induced cells expressed CK and S-100 simultaneously. The result of flow cytometry indicated that 14.1% induced cells expressed CK and S-100 simultaneously.Conclusion Induced by RPE cells lysate, rMSCs can differentiate into RPE cells.
Objective To observe the electrophysiological and morphological features of retinal ganglion cells (RGCs) in rats, and investigate its effect on the visual signal conduction. Methods Whole cell recordings were obtained from 112 RGCs of 30 rats at the age of 7-30 days. Resting membrane potential (RMP) was recorded, and input impedance was noted after given 2 mV hyperpolarizing current by voltage clamp. The action potential (AP) was induced by deplorizing current at different densities. The histological staining was actualized by injecting with biotin into the RGCs, and the diameter of the cells was measured. Results Three different discharge patterns of RGCs in response to maintained depolarizing currents were recorded: single spike (25 RGCs), transient firing (40 RGCs), and sustained firing (47 RGCs).The dia meter was 14-16μm in 57.14% transient firing RGCs, and 10-12 μm in 62.50% sustained firing RGCs. The maximum frequency of AP of sustained firing RGCs was significantly higher than that of transient firing RGCs (P＜0.05). Conclusion The single firing of RGCs was an immature electrophysiological feature. The electrophysiological features of transient firing and sustained firing RGCs may be important to make the visual information code in spatial and temporal pathway. The electrophysiological and morphological features of RGCs in rats may be correlated with each other. (Chin J Ocul Fundus Dis，2004，20：160-164)
Ophthalmic imaging examination is the main basis for early screening, evaluation and diagnosis of eye diseases. In recent years, with the improvement of computer data analysis ability, the deepening of new algorithm research and the popularization of big data platform, artificial intelligence (AI) technology has developed rapidly and become a hot topic in the field of medical assistant diagnosis. The advantage of AI is accurate and efficient, which has great application value in processing image-related data. The application of AI not only helps to promote the development of AI research in ophthalmology, but also helps to establish a new medical service model for ophthalmic diagnosis and promote the process of prevention and treatment of blindness. Future research of ophthalmic AI should use multi-modal imaging data comprehensively to diagnose complex eye diseases, integrate standardized and high-quality data resources, and improve the performance of algorithms.
Objective To elucidate the new development, structural features and appl ication of the lumbar interspinous process non-fusion techniques. Methods With the review of the development course and important research works in the field of the lumbar inter-spinous process non-fusion techniques, the regularity summary, science induction, and prospect were carried out. Results The lumbar inter-spinous process non-fusion technique was a part of non-fusion insertof spinal division posterior surface. According to the design, it could be divided into two major categories: dynamic and static systems. The dynamic system included Coflex and device for intervertebral assisted motion; the static system included X-STOP, ExtenSure and Wall is. The lumbar inter-spinous process non-fusion technique was a new technique of spinal division, it could reserve the integrated function of intervertebral disc and zygapophysial joint, maintain or recover the segmental movement to a normal level, and have no adverse effect on the neighboring segments. A lot of basic and cl inical researches indicated that lumbar inter-spinous process insert had extensive appl ication to curatio retrogression lumbar spinal stenosis, discogenic low back pain, articular process syndrome, lumbar intervertebral disc protrusion and lumbar instabil ity and so on. Conclusion With the matures of lumbar inter-spinous process non-fusion techniques and the increased study of various types of internal fixation devices, it will greatly facil itate the development of treatment of lumbar degenerative disease. But long-term follow-up is needed to investigating the long-term efficacy and perfect operation indication.
ObjectiveTo observe the effect of adenovirus-mediated Tum5 (rAd-Tum5) inhibiting retinal neovascularization (RNV) of oxygen-induced retinopathy (OIR) mouse model. MethodsThe OIR model was induced in 96 C57BL/6J mice aged of 7 days according to the literature. These mice were divided randomly into control group, OIR group, OIR rAd-green fluorescent grotein (GFP) group and OIR rAd-Tum5 group, each group had 24 mice. The rAd-GFP and rAd-Tum5 were injected into the vitreous cavity of mice aged of 12 days in OIR rAd-GFP group and OIR rAd-Tum5 group, respectively. Meanwhile, OIR group and the control group received the injection of physiological saline solution of same volume. The relatively non-perfusion area was evaluated by fluorescence angiography, and the number of pre-retinal nucleus breaking through internal limiting membranes was observed by hematoxylin-eosin staining. The expression of vascular endothelial growth factor (VEGF) was estimated by immunofluorescent (IF) and Western blot. ResultsThe retinal avascular areas of all groups were significantly different (F=61.224, P<0.01). The retinal avascular area of the rAd-Tum5 group was decreased significantly comparing with that in the OIR group and rAd-GFP group (P<0.01). However, there are no significant differences between the OIR group and rAd-GFP group (P=0.827). The number of pre-retinal nucleus breaking through ILM of all groups was significantly different (F=635.738, P<0.01), but no significantly difference was observed in OIR group and rAd-GFP group (P=0.261). Significant differences could also been seen between OIR rAd-Tum5 group and OIR group as well as OIR rAd-Tum5 group and OIR rAd-GFP group (P<0.01). The results of IF and Western blot indicated that expression of VEGF in the OIR group and rAd-GFP group was obviously up-regulated, compared with that in the control group. But the expression was declined in the rAd-Tum5 group compared with that in the OIR group and rAd-GFP group. ConclusionTum-5 peptide can efficiently prevent RNV probably by down-regulating expression of VEGF.
Objective：To investigate the role of 17beta; estradiol on th e expressi on of vascular endothelial growth factor (VEGF) and on the releasing rate of lac tate dehydrogenase (LDH) in cultured anoxiainjured human retinal pigment epit h eliual (RPE) cells. Methods:Established the anoxiainjuried m odel of human RPE c ells with Cobalt Chloride (CoCl2) after RPE cells were pretreated with 17beta;E 2 and tamoxife, 17beta;E2 antagonist. The expression of VEGF mRNA was detecte d by re v erse transcriptionpolymerase chain reaction technique (RTPCR). The cultured RP E cells were divided into four groups: normal control group, anoxiainjured gro u p, 17beta;E2 pretreatment group and 17beta;E2 with tamoxifen pretreatment grou p. The releasing rate of LDH was detected by chromatometry. The expression of VEGF pro tein were detected by cellular immunohistochemistry. Results:T he expression of VEGF and LDH releasing rate were higher in anoxiainjured grou p than that in nor m al control group (P＜0.05), and were lower in 17beta;E2 pretreatment group than th at in anoxiainjured group （P＜0.05）. When the effect of 17beta;E2 was o bstructe d by tamoxifen, the expression of VEGF and LDH releasing rate increased but didn prime;t differ much from which in anoxiainjured group (P＞0.05). Conc lusion:The ex pression of VEGF increases in anoxiainjured human RPE cells. 17beta;E2 can do wnr egulate the expression of VEGF and decrease the releasing rate of LDH, which can be blocked by tamoxifen.
Four techniques in Whipple operation improved by the anthor in this article are as follow: ①the jejunum was pulled up to the area above transverse colon through the duodenal canal behide intestinal mesenteric radix. ②As Hofmeister’s method, the duodenojejunostomy or gastrojejunostomy was made through mesentery of transverse colon. ③The internal drainage tube inserted into the pancreatic duct was extended to about 25 to 30 cm. ④A silicon tube for feeding about 3 mm diameter was placed into distal jejudum through anterior wall of gastric antrum, pylorus and duodenojejunal anastomosis. The techniques and their advantages are elaborated in this paper.
Optical coherence tomography (OCT) has developed from time-doma in into Fourier-domain OCT (FD-OCT) which indicates clearer details and higher resolution of images. FD-OCT can indicate the structure and pathological changes of each retinal layer, and reveal the retinal external limiting membranes and changes of inner- and outer-segment of visual cells by 3D solid reconstruction. FD-OCT not only provide detailed information of the images for the clinical diagnosis, but also help us investigting the characteristics and pthological mechanisms of ocular fundus diseases, which lead us to a new era of technology of observation on ocualr fundus diseases. In the application, we should pay attention to the significance of different colors of OCT images, and focus on the cohenrence of the position in the image acquistion during the follow-up period. Dynamic observation on the lesions by FD-OCT and aggregated anaylsis of resutls of several imageological examination would be the development direction of imageological examination of ocular fundus diseases.