Dysregulation and activation of immune processes are important in age-related macular degeneration (AMD) pathogenesis.The single nucleotide polymorphism of complement factor H is widely recognized as a risk factor to AMD.Over-activation of nod-like receptor3 and polymorphism of Toll-Like Receptor 3 also associated with AMD.Except for innate immune processes,adaptive immunity also play a critical role in AMD,a growing body of evidence supports that auto-antibodies and T cells are related with AMD.Additionally A2E and lipid oxidation byproducts might also have a role in AMD pathogenesis.

Objective To investigate the effects of exosomes from cultured human retinal pigment epithelium (ARPE-19) cells affected by oxidative stress on the proliferation and expression of vascular endothelial growth factor-A (VEGF-A) and Akt of ARPE-19 cells. Methods Culture ARPE-19 cells. The concentration of 2.5μmol/L rotenone was selected to simulate oxidative stress and isolated ARPE-19-exosome. Exosomes were isolated by ExoQuick exosome precipitation solution. Transmission electron microscopy was used to identify the morphology of exosomes. Western blot was used to detect exosomes’ surface-specific maker protein CD63. ARPE-19 cells affected by oxidative stress were cultured with exosome as experimental group, normal ARPE-19 cells were cultured with exosome as control group. The cell proliferation was examined by methyl thiazolyl tetrazolium assay. Western blot and immunofluorescence assay were used to detect the expression levels of VEGF-A and Akt protein. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the levels of VEGF-A mRNA and Akt mRNA. Results The diameter of normal ARPE-19-exosomes ranged from 50 to 150 nm. The isolated exosomes expressed CD63. AREP-19 cells were cultured with ARPE-19 (affected by rotenone)-exosome, the cell viability in experimental group was significantly reduced than in the control group. Green fluorescence was observed in the cytoplasm under fluorescence microscope. Compared with the control group, VEGF-A was up-regulated expressed and Akt was down-regulated expressed. Western blot results showed that, VEGF-A protein expression in the experimental group were higher than the control group. Akt protein expression in the experimental group were less than the control group. The difference was statically significant (t=3.822, 6.527;P<0.05). RT-PCR results showed that VEGF-A mRNA expression levels was higher in the experimental group than the control group. Akt mRNA expression levels was lower in the experimental group than the control group. The difference was statically significant (t=8.805,?7.823;P<0.05). Conclusions Exosomes from ARPE-19 cells affected by oxidative stress inhibit the proliferation of normal ARPE-19 cells, increase the expression of VEGF-A and reduce the expression of Akt.

Objective To investigate the effect of arginase (Arg) inhibitor N-ω-Hydroxy-L norArginine (nor-NOHA) on high glucose cultured rhesus macaque retinal vascular endothelial cell line (RF/6A) in vitro.Methods The RF/6A cells were divided into the following 4 groups:normal control group (5.0 mmol/L of glucose,group A),high glucose group (25.0 mmol/L,group B),high glucose with 125 mg/L nor-NOHA group (group C),and high glucose with 1％ DMSO group (group D).The proliferation,migration ability and angiogenic ability of RF/6A cells were measured by Methyl thiazolyl tetrazolium (MTT),transwell chamber and tube assay respectively.The express of Arg Ⅰ,eNOS,iNOS mRNA of RF/6A cells were measured by real-time polymerase chain reaction (RT-PCR),Enzyme-linked immuno sorbent assay (ELISA) was used to detect the expression of NO and interleukine (IL)-1b of RF/6A cells.Results The proliferation,migration,and tube formation ability of group A (t=2.367,5.633,7.045;P＜0.05) and group C (t=5.260,6.952,8.875;P＜0.05)were significantly higher than group B.RT-PCR results showed the Arg Ⅰ and iNOS expression in group B was higher than that in group A (t=6.836,3.342;P＜0.05) and group C (t=4.904,7.192;P＜0.05).The eNOS expression in group B was lower than that in group A and group C (t=4.165,6.594;P＜0.05).ELISA results showed NO expression in group B was lower than that in group A and group C (t=4.925,5.368;P＜0.05).IL-1b expression in group B was higher than that in group A and group C (t=5.032,7.792;P＜0.05).Conclusions Nor-NOHA has a protective effect on cultured RF/6A cells in vitro and can enhance its proliferation,migration and tube formation.The mechanism may be inhibiting the oxidative stress by balancing the expression of Arg/NOS.

Humans ; Nasal Cavity ; pathology ; Nose Neoplasms ; diagnosis ; Paranasal Sinuses ; Solitary Fibrous Tumors ; diagnosis

Objective To observe the image characteristics ofmultispectral scanning laser imaging (MSLI) and OCT in patients with pregnancy induced hypertension syndrome (PIHS).Methods A total of 112 patients (224 eyes) of PIHS patients diagnosed in Obstetrics Department of Tianjin First Central Hospital from May 2016 to May 2017 were included in this study.The average age of the patients was 27.00±2.14 years.The average course of the disease was 15.00 ±8.27 days.There were 174 eyes in 87 patients of blurred vision,dazzling and visual fatigue consciously.All patients performed BCVA,direct ophthalmoscope,B ultrasound,confocal scanning laser Ophthalmoscope (cSLO) and spectral-domain OCT (SD-OCT).SD-OCT was performed with Spectralis HRA+OCT from Heidelberg Company in Germany to acquire tomographic images.Using Herdelberg's colorful program (MultiColor) based on cSLO and operating in accordance with standard methods,one scan simultaneously obtained blue light reflection based on 488 nm,green light reflection based on 515 nm,and infrared reflection based on 820 nm,synthesis to MSLI.Fundus abnormalities were classified into arterial spasm (stage Ⅰ),arteriosclerosis (stage Ⅱ),and retinopathy (stage Ⅲ).OCT examination was divided into normal and abnormal cases according to the abnormality of retinal morphology and thickness.Results Of the 224 eyes,68 eyes (30.36％) showed normal fundus examination and 156 eyes (69.64％) showed abnormal fundus performance.Among them,28 eyes were stage Ⅰ (17.95％);40 eyes were stage Ⅱ (25.64％);88 eyes were stage Ⅲ (56.41％).Thirty-six eyes (16.07％) showed normal fundus and 188 eyes (83.93％) showed abnormal performance with OCT.Of the 188 eyes with abnormal fundus performance,86 eyes (45.74％) had retinal neuroepithelial serous detachment;56 eyes (29.79％) had RPE detachment;optic disc edema,bulge,and local reflexes in the retinal nerve fiber layer were enhanced and/or the thickness increased in 46 eyes (24.47％).In MSLI,48 eyes (21.43％) showed normal fundus;176 eyes (78.57％) showed abnormal performance.Retinal edema was showed in green on MSLI,serous retinal neuroepithelial layer detachment,RPE layer detachment,retinal nerve fiber layer thickening,accompanied by changes in local retinal structure.The higher the degree of bulge,the darker the color.Consistent with the range of retinal edema revealed by SD-OCT.Conclusions MSLI and SD-OCT images show highly consistent lesions in PIHS patients.MSLI can more clearly show superficial and deep retinal lesions.

Objective To observe the effect of exosomes secreted by retinal pigment epithelial (RPE) cells which damaged by blue light to Nod-like receptor protein (NLRP3). Methods Cultured ARPE-19 cells were divided into 2 groups; one group of RPE cells were exposed to blue light irradiation for 6 hours, the other group was cultured in routine environment. Total exosomes were extracted from the two groups by differential ultracentrifugation in low-temperature, and examined by transmission electron microscope to identify their forms. The exosomes were then incubated with normal ARPE-19 cells. The expression level of CD63, interleukin (IL)-1β, IL-18 and caspase-1 on the exosome surface were measured by Western blotting. The expressions of NLRP3 mRNA in RPE cells were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR). Results Blue light damaged the cellular morphology. Transmission electron microscopy showed that the exosomes were 50-200nm in diameter and like double-concave disks. Blue light damaged cell-derived exosomes had significantly higher expression of IL-1β (t=18.04),IL-18 (t=12.55) and caspase-1 (t=14.70) than the control group (P<0.001). ARPE-19 cells cultured with blue light damaged cell-derived exosomes also had significantly higher expression of IL-1β (t=18.59), IL-18 (t=23.95) and caspase-1 (t=35.27) than control exosomes (P<0.001). RT-PCR showed that the relative expression of NLRP3 mRNA of PRE cells in experimental group and control group were 1.000±0.069 and 0.2±0.01, respectively, the difference was significant (t=12.20, P<0.001). Conclusion The expression IL-1β, IL-18 and caspase-1 and NLRP3 mRNA were upregulated by exosomes secreted by blue light damaged-RPE cells.

Objective: To compare the clinical effects of two different skin preparation methods for infant craniocerebral surgery. Methods: Totally 120 infants who were going to receive craniocerebral surgery were divided into two groups by random number table, 60 cases in the observation group and 60 cases in the control group. The scalp of both groups was cleaned with moisturizing oil every day from 3 days before operation. On 1 day before operation, the observation group used electric shaver to shave off all hair on the head, and then rinsed with warm water. The control group was treated with skin preparation knife to shave all the hair under soap water lubrication and rinse with warm water. The skin injury rate, incision infection rate and pain score of the two groups were evaluated. Results: The incidence of skin injury and incision infection were 0 and 1.7% (1/60) in the observation group, 18.3% (11/60) and 13.3% (8/60) in the control group, respectively. There were significant differences between the two groups (χ²= 12.110, 5.886, all P < 0.01 or 0.05). The median score of pain in the observation group was 0 (Q₁:0, Q₃:0), while 1.5 (Q₁:1, Q₃:2) in the control group, and there were significant differences between the two groups (Z= 3.286, P < 0.01). Conclusion Electric shaver is superior to skin preparation knife in shaving hair of infants. It not only reduces the incidence of head skin injury, incision infection and pain in the process of skin preparation, but also reduces the incidence of incision infection after craniocerebral surgery in infants. It is worth popularizing.

Objective To compare the test performance of different immunoassays for the detection on autoantibodies specific to primary biliary cholangitis,including anti-mitochondrial type 2 antibody(AMA-M2),anti-glycoprotein 210(anti-gp210)and anti-nuclear body protein sp100(anti-sp100).Methods Serum samples from Primary Biliary Cholangitis(PBC, n=91), liver disease control(including viral hepatitis,autoimmune hepatitis and liver cirrhosis,n=67)and healthy individual(n=40)were collected from Beijing Youan Hospital during the period between April 2014 and April 2017.All samples were tested with chemiluminescent immunoassay(CLIA)and enzyme linked immunosorbent assay(ELISA)for AMA-M2, meanwhile the detection on anti-gp210 and anti-sp100 were compared between CLIA and Line Immunoassay(LIA).The Kappa coefficient were used to measure the level of qualitative agreement between different assays.The diagnostic accuracy of AMA-M2 detected with CLIA and ELISA were compared by receiver operating characteristic curve(ROC).Results The overall qualitative agreement between CLIA and ELISA for the detection to AMA-M2 is 88.4%(Kappa =0.765, P<0.01).Excellent qualitative agreement between CLIA and LIA for the detection to anti-gp210 and anti-sp100 was also found with overall agreement as 96.5%(Kappa=0.852,P<0.01)and 98%(Kappa=0.884,P<0.01), respectively.The ROC analysis also showed similar area under the curve(AUC)for CLIA(0.965, P<0.01)and ELISA (0.928,P<0.01)on detection to AMA-M2.Conclusions CLIA and ELISA showed excellent agreement for the detection to AMA-M2.High qualitative agreement between CLIA and LIA was also found when testing anti-gp210 and anti-sp100.

Coactosin-like protein 1 (Cotl1), a member of the actin-depolymerizing factor (ADF)/cofilin family, was first purified from a soluble fraction of Dictyostelium discoideum cells. Neuronal migration requires cytoskeletal remodeling and actin regulation. Although Cotl1 strongly binds to F-actin, the role of Cotl1 in neuronal migration remains undescribed. In this study, we revealed that Cotl1 overexpression impaired migration of both early- and late-born neurons during mouse corticogenesis. Moreover, Cotl1 overexpression delayed, rather than blocked, neuronal migration in late-born neurons. Cotl1 expression disturbed the morphology of migrating neurons, lengthening the leading processes. This study is the first to investigate the function of Cotl1, and the results indicate that Cotl1 is involved in the regulation of neuronal migration and morphogenesis.
Actins ; Animals ; Dictyostelium ; Humans ; Mice ; Morphogenesis ; Neurons

Objective:To investigate the prevalence of small intestinal bacterial overgrowth (SIBO) in patients with rosacea, and to analyze the relationship between breath test results and the occurrence of rosacea.Methods:Patients with rosacea were enrolled from the outpatient department of Xiangya Hospital from March 2022 to June 2023. The methane-hydrogen breath test was used to detect intestinal levels of methane and hydrogen in all patients to investigate the prevalence of SIBO. The basic information, clinical symptoms and severity, quality of life scores, gastrointestinal symptoms, and past medical history of the patients were collected. Statistical analysis was carried out by using the chi-square test, nonparametric test and multivariate logistic regression models to investigate the relationship between SIBO and the occurrence of rosacea.Results:A total of 116 patients with rosacea completed the methane-hydrogen breath test. They were aged 18 to 56 years (median [ Q1, Q3]: 25 [22, 33] years), and included 7 males (6.0%) and 109 females (94.0%) ; there were 43 cases (37.1%) of erythematotelangiectatic rosacea, and 73 (62.9%) of papulopustular rosacea. As the breath test showed, 94 patients were diagnosed with SIBO (81.0%, 95% CI: 72.7% - 87.7%) based on the breath tests, 84 showed positive hydrogen breath test results (72.4%, 95% CI: 63.3% - 80.3%), and 47 had positive methane breath test results (40.5%, 95% CI: 31.5% - 50%). Among the 67 patients with moderate to severe erythema, 33 (49.3%) showed positive methane breath test results, and 14 of 49 (28.6%) patients with mild erythema showed positive methane breath test results, with a rate difference of 20.7% ( P = 0.025, 95% CI: 13.9% - 27.5%) ; there were no significant differences in the positive rates of SIBO and hydrogen breath test results between the patients with moderate to severe erythema and those with mild erythema (both P > 0.05). No significant differences were observed in the age, gender, clinical subtypes, severity of papulopustules, flushing and burning sensation, or rosacea quality of life index scores between the SIBO-positive and -negative groups, between hydrogen-positive and -negative groups, and between methane-positive and -negative groups (all P > 0.05). Multivariate logistic regression analysis showed that methane positivity on breath test was associated with the severity of erythema in rosacea ( OR = 2.495, 95% CI: 1.102 - 5.649, P < 0.05) . Conclusions:The prevalence of SIBO was relatively high in the patients with rosacea. However, only the positive rate of methane breath test differed between the rosacea patients and non-rosacea controls, and there was some correlation between methane positivity on breath test and increased severity of rosacea erythema.