ObjectiveTo investigate the inhibitory effect of lentivirus-mediated polypyrimidine bundle binding protein-associated splicing factor (PSF) on retinal neovascularization (RNV) in mice model of oxygen-induced retinopathy (OIR).MethodsOne hundred and twelve 5-day-old C57BL/6J mice were randomly divided into normal control group, simple OIR model group, OIR model + lentivirus empty vector treatment group (Vec group) and OIR model + PSF lentivirus treatment group (PSF group), with 16, 32, 32 and 32 mice, respectively. When the mice were 7 days old, the mice in the normal control group were fed in a routine environment, and the mice in the OIR model group, Vec group and PSF group were established OIR model. The mice in the Vec group and PSF group were given an intravitreal injection of 1 μl of lentiviral vector and PSF lentivirus (titer 1×1011 TU/ml) at the age of 12 days. No injection was performed in the normal control group and simple OIR group. RNV was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area by immunofluorescent staining of the mouse retina. Real-time quantitative PCR was applied to detect the mRNA expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and hemeoxygenase-1 (HO-1). Western blot analysis was applied to detect the protein expression of Nrf2, HO-1 and PSF. Results Of the normal control group, simple OIR model group, Vec group and PSF group, the number of pre-retinal neovascular cell nuclei were 0.00, 14.36±5.50, 15.67±4.96, 8.13±2.09, the non-perfusion area were 0.00%, (35.71±2.81)%, (36.57±4.53)%, (15.33±4.75)%, respectively. The differences of the number of pre-retinal neovascular cell nuclei and non-perfusion area among 4 groups were significant (F=24.87, 165.70; P＜0.05). Compared with the normal control group, there were more pre-retinal neovascular cell nucleis and larger non-perfusion area in the simple OIR model group and Vec group (P＜0.05). Compared with the simple OIR model group and Vec group, there were lower pre-retinal neovascular cell nucleis and smaller non-perfusion area in the PSF group (P＜0.05). Real-time quantitative PCR and Western blot showed that the mRNA expression of Nrf2, HO-1 (F=53.66, 83.54) and protein expression of Nrf2, HO-1 and PSF (F=58.38, 52.69, 24.79) among 4 groups were significant (P＜0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the simple OIR model group and Vec group decreased significantly than those in the normal control group (P＜0.05). The mRNA expression of Nrf2, HO-1 and protein expression of Nrf2, HO-1 and PSF in the PSF group were increased significantly than those in the simple OIR model group and Vec group (P＜0.05). model group and Vec group (P＜0.05).ConclusionIntravitreal injection of lentivirus-mediated PSF inhibits RNV in mice model of OIR possibly through up-regulating the expression of Nrf2 and HO-1.
目的 探討胃癌組織中人表皮生長因子受體（EGFR）、表皮生長因子受體-2（HER-2）和環氧合酶-2（COX-2）的表達及與臨床病理特征的關系。 方法 應用免疫組織化學Envision二步法，檢測70例胃癌組織中EGFR、HER-2和COX-2的表達情況，并結合其臨床病理特點進行分析。 結果 EGFR、HER-2和COX-2在胃癌組織中的表達陽性率分別為35.7%（25/70）、27.1%（19/70）、67.1%（47/70）。陽性表達與腫瘤分化程度、侵襲深度、有無淋巴結轉移及TNM分期有關（P＜0.05），而與患者的性別及年齡、腫瘤部位和大小無關（Pgt;0.05）。EGFR、HER-2和COX-2三者之間在胃癌組織中的表達均呈正相關（P＜0.05）。 結論 EGFR、HER-2和COX-2的表達參與胃癌的生長、侵襲和轉移過程。它們的聯合檢測有助于胃癌患者靶向藥物的選擇，也為胃癌的預后判斷提供客觀的參考指標。Objective To observe the expressions of epidermal growth factor receptor (EGFR), HER-2 and cyclooxygenase-2 (COX-2) in gastric carcinoma (GC) and to explore the relationship among them. Methods The envision immunohistochemical stain method was used to detect EGFR, HER-2 and COX-2 protein expressions in sample of 70 GC tissues. And their corresponding pathologic features were analyzed. Results The positive expression rates of EGFR, HER-2 and COX-2 protein in GC tissue were 35.7% (25/70), 27.1% (19/70) and 67.1% (47/70), respectively. The positive expression rates were closely relevant to the differentiation of the cancer, invasion depth, lymphatic metastasis and TNM (P＜0.05), but not to the patient’ s sex, age, tumor site and size (P＞0. 05). There was a stable positive correlation among EGFR, HER-2 and COX-2 expressions in GC tissues, respectively. Conclusions EGFR, HER-2 and COX-2 expressions participate in the development, invasion and metastasis process of GC. Combined detection can be regarded as an important symbol for guiding the molecular targeting therapy of GC, and judging the prognosis of GC.
Objective
To study the relationship between the expression ratio of induced nitric oxide synthase (iNOS) over glial fibrillary acidic protein (GFAP) and the time of injury after brain concussion in rat, in order to acquire a new visual angle for determining injury time of cerebral concussion.
Methods
Eighty-five healthy Sprague-Dawley rats were divided into three groups randomly: model group (n=25), experimental group (n=55), and control group (n=5). The rats in the model group were used to confirm the attack hight to make the model of brain concussion; according to the time of execution, rats in the experimental group were then subdivided into 11 groups with 5 rats in each subgroup, and their execution time was respectively hour 0.5, 1, 3, 6, 12, 24, 48, 96, 168, 240, and 336; the rats in the control group were executed after fed for 24 hours. After the model of cerebral concussion was established through freefalling dart method, hematoxylin-eosin staining and immunohistochemistry staining of iNOS and GFAP were conducted for the brain of the rats. All related experimental results were studied by using microscope with image analytical system and homologous statistics.
Results
The ratio of positive expression of iNOS over that of GFAP increased gradually during hour 0.5- 3 after injury in brain (from 5.03 to 10.47). At the same time, the positive expression of iNOS increased significantly (from 14.61% to 37.45%). However, the increase of the positive expression of GFAP was not obvious. Between hour 3 and 12, the ratio began to decline to 4.98, which was still at a high level, and during the same time period, the positive expressions of iNOS and GFAP also experienced the same change pattern. Later, the ratio began to decline between hour 12 and 336 after injury (from 4.98 to 0.95). All ratios at this time were lower than those between hour 0.5 and 12. The positive expression of iNOS and GFAP both increased to a climax before declining.
Conclusions
The ratio of positive expression of iNOS over GFAP and the respective change pattern of iNOS and GFAP can be used as the evidence of estimating the injury time of cerebral concussion. We can use the ratio of two or more markers to provide a new visual angle for concluding the concussion injury time.
ObjectiveTo study a deep learning-based dual-modality fundus camera which was used to study retinal blood oxygen saturation and vascular morphology changes in eyes with branch retinal vein occlusion (BRVO). MethodsA prospective study. From May to October 2020, 31 patients (31 eyes) of BRVO (BRVO group) and 20 healthy volunteers (20 eyes) with matched gender and age (control group) were included in the study. Among 31 patients (31 eyes) in BRVO group, 20 patients (20 eyes) received one intravitreal injection of anti-vascular endothelial growth factor drugs before, and 11 patients (11 eyes) did not receive any treatment. They were divided into treatment group and untreated group accordingly. Retinal images were collected with a dual-modality fundus camera; arterial and vein segments were segmented in the macular region of interest (MROI) using deep learning; the optical density ratio was used to calculate retinal blood oxygen saturation (SO2) on the affected and non-involved sides of the eyes in the control group and patients in the BRVO group, and calculated the diameter, curvature, fractal dimension and density of arteriovenous in MROI. Quantitative data were compared between groups using one-way analysis of variance. ResultsThere was a statistically significant difference in arterial SO2 (SO2-A) in the MROI between the affected eyes, the fellow eyes in the BRVO group and the control group (F=4.925, P<0.001), but there was no difference in the venous SO2 (SO2-V) (F=0.607, P=0.178). Compared with the control group, the SO2-A in the MROI of the affected side and the non-involved side of the untreated group was increased, and the difference was statistically significant (F=4.925, P=0.012); there was no significant difference in SO2-V (F=0.607, P=0.550). There was no significant difference in SO2-A and SO2-V in the MROI between the affected side, the non-involved side in the treatment group and the control group (F=0.159, 1.701; P=0.854, 0.197). There was no significant difference in SO2-A and SO2-V in MROI between the affected side of the treatment group, the untreated group and the control group (F=2.553, 0.265; P=0.088, 0.546). The ophthalmic artery diameter, arterial curvature, arterial fractal dimension, vein fractal dimension, arterial density, and vein density were compared in the untreated group, the treatment group, and the control group, and the differences were statistically significant (F=3.527, 3.322, 7.251, 26.128, 4.782, 5.612; P=0.047, 0.044, 0.002, <0.001, 0.013, 0.006); there was no significant difference in vein diameter and vein curvature (F=2.132, 1.199; P=0.143, 0.321). ConclusionArterial SO2 in BRVO patients is higher than that in healthy eyes, it decreases after anti-anti-vascular endothelial growth factor drugs treatment, SO2-V is unchanged.
Objective To explore effects of edaravone on apoptosis and expressions of apoptotic proteins Smac and XIAP in hippocampal CA1 pyramidal cell of rats under intermittent hypoxia. Methods A total of 96 adult male Wistar rats were randomly divided into control group, 5% intermittent hypoxic group and edaravone group, and each group was divided into 4 time groups at 7 d, 14 d, 21 d and 28 d, respectively, with 8 rats in each subgroup. The content of reactive oxygen species (ROS) in hippocampal tissues of the experimental rats was detected by the reactive oxygen species detection kit. Immunohistochemistry and Western blot were used to detect the expressions of Smac and XIAP protein in hippocampal CA1 region. The Tunel method detected the apoptosis of neurons. Results Compared with the control group, the content of ROS, the expressions of Smac and XIAP proteins and the neuronal apoptosis index in the hippocampus were increased in the 5% intermittent hypoxia group and the edaravone group at each time point (all P<0.05). The content of ROS, the Smac protein expression and the neuronal apoptosis index in the edaravone group were significantly lower than those in the 5% intermittent hypoxia group (all P<0.05). The expression of XIAP protein in the edaravone group was significantly higher than that in the 5% intermittent hypoxia group (P<0.05). Conclusion Edaravone may improve the antioxidant capacity of the body by scavenging oxygen free radicals and regulate Smac and XIAP- mediated apoptosis, thus playing a protective role on neurons.
Objective To investigate the cellular viability and mitochondrial reactive oxygen species (ROS) production of the Müller cells under high glucose condition, and explore the protection role of the 5,6-dihydrocyclopenta-1, 2-dithiole-3-thione (CPDT) on Müller cells. Methods Müller cells from Sprague Dawley rats were divided into 5 groups randomly, including 25 mmol/L normal glucose group (group A) and 65 mmol/L high glucose group (group B). High glucose group with 45, 60, 70 μmol/L CPDT and cultured them 72 hour was set as group C, D and E. Water soluble tetrazolium salt (WST)-8 was used to measure the cellular viability. Flow cytometry was used to measure the active oxygen and apoptosis index. The expression of nuclear factor erythroid 2-related factor 2 (Nrf2), hemeoxygenase-1 (HO-1), Bcl-2 and Bax protein were measured by Western blot. Results Compared with group A, the WST-8 showed that the viability of Müller cells apparently decreased in group B (t=39.59,P＜0.05). Compared with the group B, the viability of Müller cells had changes in group C (t=0.97,P＞0.05), but recovered in group D and E (t=?4.17, ?7.52;P＜0.05). Compared with group A, the FCM showed that the mitochondrial ROS levels was higher in group B (t=?30.99,P＜0.05). Compared with group B, the mitochondrial ROS levels were decreased in group D (t=27.68,P＜0.05). Compared with group A, Bax, Nrf2 and HO-1 increased (t=–11.03, –63.17, –11.44;P＜0.05), while the bcl-2 decreased in group B (t=7.861,P＜0.05). Compared with the group B, Nrf2, HO-1 and Bax decreased (t=15.11, 26.59, 6.27;P＜0.05), while the bcl-2 increased in group D (t=?6.53,P＜0.05). Conclusions Under the high glucose, CPDT may reduce the mitochondrial ROS levels and the expression of Nrf2, HO-1 and Bax protein of Müller cells. It may inhibit apoptosis through activating the Nrf2/HO-1 pathway and balancing of level of Bcl-2 protein and mitochondrial ROS.
