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Objective:To observe and preliminarily explore the effect of mogroside on oxidative stress of retinal pigment epitheliaum (RPE) cells induced by hydrogen peroxide (H 2O 2) and its possible mechanism. Methods:A experimental study. The RPE cells were divided into control group, H 2O 2 group, silent information regulator of transcription 1 (SIRT1) inhibitor EX527 group (EX527 group), mogroside group, mogroside+EX527 group. Methyl thiazolete trazolium method was used to detect cell survival rate. Flow cytometry was used to detect cell apoptosis rate. 2' ,7'-dichlorodihydrofluorescein diacetate fluorescent probe method, xanthine method and enzyme-linked immunosorbent assay method were used to detect the level of reactive oxygen species (ROS), superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cells respectively. Real-time quantitative polymerase chain reaction and Western blot were used to detect relative expressions of SIRT1, nuclear factor erythroid-2-related actor 2 (Nrf2), heme oxygenase-1 (HO-1) mRNA and protein in cells. One-way ANOVA was used for comparison among groups. The pairwise comparison between groups was tested by the least significant difference t test. Results:Compared with the control group, the H 2O 2 group cell survival rate decreased, the apoptosis rate increased, the ROS level in the cells increased, the SOD activity decreased, the MDA content increased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein decreased ( P<0.05). Compared with H 2O 2 group, the cell survival rate decreased, apoptosis rate increased, the cell ROS level increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein expression decreased in EX527 group ( P<0.05); the cell survival rate increased, apoptosis rate decreased, ROS level decreased, SOD activity increased, MDA content decreased, and the relative expression of SIRT1, Nrf2, HO-1 mRNA and protein increased in mogroside group ( P<0.05). Compared with the mogrosides group, the cell survival rate decreased, the apoptosis rate increased, the level of ROS increased, SOD activity decreased, MDA content increased, SIRT1, Nrf2, HO-1 mRNA and protein decreased in mogrosides+EX527 group ( P<0.05). Conclusions:Mogrosides can alleviate the oxidative stress response of visual RPE cells induced by H 2O 2, promote cell proliferation, and reduce cell apoptosis. Mogrosides may exert antioxidant effects by activating the SIRT1/Nrf2 signaling pathway.
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Objective To investigate the role of methyl-CpG-binding protein 2 (MeCP2) in the regulation and epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs) and its possible mechanism.Methods Human LEC lines (SRA01/04) were divided into MeCP2-mimic group,MeCP2-NC group and small interferening RNA-MeCP2 (si-MeCP2) group,and MeCP2 analog plasmid,blank plasmid and MeCP2 si-RNA plasmid was used respectively to transfect the cells.The expression of MeCP2 mRNA in the cells was detected by using real-time PCR 24 hours after transfection.At 48 hours after transfection,the migration rate of the cells was evaluated by scratching test,and the expression of Wnt3a protein in the cells was detected by immunofluorescence stainning.The relative expressions of β-catenin,E-cadherin,Vimentin,matrix metallo proteinase (MMP)-9,MMP-7 and secreted frizzled-related protein 5 (SFRP5) proteins in the cells were detected by Western blot.Results After 24 hours of transfection,the relative expression of MeCP2 mRNA in the cells was significantly different among the MeCP2-mimic group,MeCP2-NC group and si-MeCP2 group (F =4 773.00,P＜0.00 1).The migrating rate of the cells in the MeCP2-mimic group,MeCP2-NC group and si-MeCP2 group was (57.45±5.20)％,(32.71± 10.02)％ and (17.77±9.22)％,respectively,showing a significant difference among the three groups (F=124.00,P＜0.001),and the migrating rate of the cells in the si-MeCP2 group was significantly lower than that of the MeCP2-mimic group or MeCP2-NC group (both at P＜0.001).The relative expressing intensity (absorbance) of Wnt3a in the cells of the MeCP2-mimic group,MeCP2-NC group and si-MeCP2 group was 75.92 ± 6.10,52.03 ± 5.22 and 28.75 ± 3.39,respectively,with a significant difference among three the groups (F=221.30,P＜0.001),and the relative expressing intensity (absorbance) of Wnt3a in the cells was significantly lower in the si-MeCP2-mimic group than that of the MeCP2-NC group and MeCP2-mimic group (both at P＜0.001).The relative expressing level of E-cadherin protein was significantly elevated and the expressions of β-catenin,Vimentin,MMP-9 and MMP-7 were significantly reduced in the si-MeCP2 group compared with the MeCP2-mimnic group and MeCP2-NC group (all at P＜O.01).The relative expressing level of SFRP5 protein in the MeCP2-mimic group,MeCP2-NC group and si-MeCP2 group was 27.19± 0.03,47.54±0.05 and 74.93±0.05,respectively,showing a statistical difference among the three groups (F =183.49,P＜0.001),and the relative expressing level of SFRP5 in the si-MeCP2 group was significantly higher than that in the MeCP2-mimic group and MeCP2-NC group (both at P＜0.001).Conclusions MeCP2C can promote EMT of human LECs by down-regulating the expression of SFRP5 and therefore activating the Wnt3a/β-catenin signal pathway.
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Kawasaki disease is an acute systemic vascular inflammatory syndrome, which is the main cause of acquired heart disease in children.It is most likely to cause serious complications such as coronary artery dilation, coronary artery aneurysm, and acute myocardial infarction.At present, there is no accurate conclusion about the etiology and pathogenic mechanism of Kawasaki disease.We performed this review to realize the etiology of Kawasaki disease regarding infectious pathogens, environmental factors, immune disorders and genetic tendencies.Meanwhile, this study will focus on the abnormal activation of the immune system, the up-regulated expression of inflammatory cytokines, the increased activity of metal matrix proteinases (MMPs), and vascular endothelial injury/vascular endothelial dysfunction to review the pathogenic mechanism of Kawasaki disease.
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Objective@#To investigate the role of methyl-CpG-binding protein 2 (MeCP2) in the regulation and epithelial-mesenchymal transition (EMT) of human lens epithelial cells (LECs) and its possible mechanism.@*Methods@#Human LEC lines (SRA01/04) were divided into MeCP2-mimic group, MeCP2-NC group and small interferening RNA-MeCP2 (si-MeCP2) group, and MeCP2 analog plasmid, blank plasmid and MeCP2 si-RNA plasmid was used respectively to transfect the cells.The expression of MeCP2 mRNA in the cells was detected by using real-time PCR 24 hours after transfection.At 48 hours after transfection, the migration rate of the cells was evaluated by scratching test, and the expression of Wnt3a protein in the cells was detected by immunofluorescence stainning.The relative expressions of β-catenin, E-cadherin, Vimentin, matrix metallo proteinase (MMP)-9, MMP-7 and secreted frizzled-related protein 5 (SFRP5) proteins in the cells were detected by Western blot.@*Results@#After 24 hours of transfection, the relative expression of MeCP2 mRNA in the cells was significantly different among the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group (F=4 773.00, P<0.00 1). The migrating rate of the cells in the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was (57.45±5.20)%, (32.71±10.02)% and (17.77±9.22)%, respectively, showing a significant difference among the three groups (F=124.00, P<0.001), and the migrating rate of the cells in the si-MeCP2 group was significantly lower than that of the MeCP2-mimic group or MeCP2-NC group (both at P<0.001). The relative expressing intensity (absorbance) of Wnt3a in the cells of the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was 75.92±6.10, 52.03±5.22 and 28.75±3.39, respectively, with a significant difference among three the groups (F=221.30, P<0.001), and the relative expressing intensity (absorbance) of Wnt3a in the cells was significantly lower in the si-MeCP2-mimic group than that of the MeCP2-NC group and MeCP2-mimic group (both at P<0.001). The relative expressing level of E-cadherin protein was significantly elevated and the expressions of β-catenin, Vimentin, MMP-9 and MMP-7 were significantly reduced in the si-MeCP2 group compared with the MeCP2-mimnic group and MeCP2-NC group (all at P<0.01). The relative expressing level of SFRP5 protein in the MeCP2-mimic group, MeCP2-NC group and si-MeCP2 group was 27.19±0.03, 47.54±0.05 and 74.93±0.05, respectively, showing a statistical difference among the three groups (F=183.49, P<0.001), and the relative expressing level of SFRP5 in the si-MeCP2 group was significantly higher than that in the MeCP2-mimic group and MeCP2-NC group (both at P<0.001).@*Conclusions@#MeCP2C can promote EMT of human LECs by down-regulating the expression of SFRP5 and therefore activating the Wnt3a/β-catenin signal pathway.
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OBJECTIVE@#To study the clinical features of children with bronchial asthma complicated by pulmonary fungal infection and the risk factors for pulmonary fungal infection.@*METHODS@#A retrospective analysis was performed for the clinical data of 150 children with bronchial asthma who were admitted from January 2015 to June 2018. Among these children, 75 had pulmonary fungal infection (fungal infection group) and 75 did not have such infection (control group). The distribution of pathogenic fungi, clinical symptoms/signs and treatment outcome were recorded for the fungal infection group. The multivariate logistic regression analysis was used to investigate the risk factors for pulmonary fungal infection.@*RESULTS@#A total of 69 pathogenic fungi were detected in 75 children in the fungal infection group, among which Candida albicans had the highest detection rate of 61%. Major clinical symptoms were cough (93%), persistent high fever (56%), wheezing (49%) and dyspnea (48%). Major signs were dry and moist rales (43%) and moist rales (29%). Parts of children had hepatosplenomegaly. Among the 75 children in the fungal infection group, 39 were markedly improved, 26 were improved, 7 had no response, and 3 experienced aggravation and then died. Age 3 times during hospitalization, intravenous administration of glucocorticoids, non-rational use of antibiotics, mechanical ventilation and prolonged hospital stay were independent risk factors for pulmonary fungal infection in children with asthma (OR=4.865, 3.241, 2.255, 3.725, 3.568, 1.549, 3.808; P3 times during hospitalization, intravenous administration of glucocorticoids, non-rational use of antibiotics, mechanical ventilation or prolonged hospital stay have a higher risk for secondary pulmonary fungal infection.
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Child , Humans , Asthma , Lung Diseases, Fungal , Retrospective Studies , Rhinitis, Allergic , Risk FactorsABSTRACT
Objective To explore the clinical characteristics of systemic disseminated infection caused by Mycobacterium fortuitum (M.fortuitum), and improve the diagnostic rate and understanding of the disease.Methods One case of systemic disseminated M.fortuituminfection was reported, and analyzed in combination with relevant literatures.Results Patient was with multiple systemic involvement (including lung, lymph node, skin, joint), lymph node tissue culture was positive for M.fortuitum, patient was given clarithromycin+levofloxacin+linezolid for treatment, disease was remitted.Conclusion Systemic disseminated M.fortuituminfection is rare, and patient with GATA2 deletion and IFN-γautoantibody may be a potential mechanism, diagnosis is mainly based on pathological morphology and microbiological detection, but positive rate is low, diagnosis is difficult.
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Background Congenital stationary night blindness (CSNB) is a common genetic eye disease.Retinal angiogenesis is rarely obtained in retinal degeneration animal.The effects of CSNB on retinal angiogenesis require further study.Objective This study was to improve Chinese ink perfusion technology and to explore the effect of CSNB on oxygen induced neovascularization.Methods Eighteen clean 7-day old SD rats and 18 clean 7-day old CSNB rats were included,twelve SD rats and twelve CSNB rats were chosen randomly for oxygen induced retinopathy (OIR) modeling,and served as OIR group,six SD rats and six CSNB rats were chosen as normal control.Nine rats were chosen randomly from both SD rats and CSNB rats in OIR group,respectively.The rats were separated into 1 ∶ 1 ratio ink group,2 ∶ 1 ratio ink group and conventional ink group,which were perfused with 1 ∶ 1 ratio ink perfusate,2 ∶ 1 ratio ink perfusate and conventional ink,respectively.The unilateral eyes of the rats were prepared for whole-mount retina,the other eyes were performed for paraffin imbedding.The quality of retinal vascular imaging were compared among different ink perfusate groups.The normal control rats,three SD rats in OIR group and three CSNB rats in OIR group were perfused with 2 ∶ 1 ratio ink perfusate.Histopathology examination was performed on the paraffin section,and the number of nuclei breakthrough the inner limiting membrane were counted.Immunocytochemistry were performed on the paraffin section for detecting the expression of von Wilebrand factor (vWF).Results Compared with 1 ∶ 1 ratio ink perfusion and conventional ink perfusion,2 ∶ 1 ratio ink perfusion showed the full vascular net clearly.Histopathology showed that the structure of retina was normal in the normal control group,and there were no endothelial nuclei breakthrough the inner limiting membrane.A large number of endothelial nuclei breakthrough the inner limiting membrane in the OIR group,the number of endothelial nuclei breaking through the inner limiting membrane were (23.08±2.99)/slide and (41.12±9.36)/slide for SD rats and CSNB rats,respectively.The number of endothelial nuclei breakthrough inner limiting membrane was higher in the CSNB rats than that in the SD rats,with no difference between the two group (q =1.70,P =0.50).Immunocytochemistry results showed that v-WF was positive expressed in the cells breakthrough inner limiting membrane.Conclusions Improved ink perfusion method was an easy-to-use whole-mount retina method with good repeatability.Hyperoxia can induce retinal neovascularization of CSNB rats.
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<p><b>OBJECTIVE</b>To study the mechanism of learning and memory dysfuction in the transgenic mouse expressing human tau 40 isoform with P301L mutation (F10).</p><p><b>METHODS</b>The human tau protein expression and phosphor-tau protein levels were detected with Western blot method. The neurofibrillary tangles were observed with Bielshowsky silver stain. The behavior changes of learning and memory were observed by open field test and passive avoidance test. Acetyleholine level, activities of acetycholinesterase and choline acetyltransferase of whole brain was detected by colorimetry method. The nitric oxide level of whole brain was detected by nitrate enzyme reduction method.</p><p><b>RESULTS</b>Exogenous human tau gene was expressed and an elevation of phosphor-tau protein level in 7 and 3-month transgenic mice's hippocampus andcerebrocortex was observed. The neurofibrillary tangles were observed in cerebrocortex of 7-month transgenic mice; the 7-month transgenic mice also presented an evident reduction of learning and memory ability and nitric oxide level of the whole brain, but not changes in acetylcholine level, acetycholinesterase activity, choline acetyltransferase activity and expression in whole brain.</p><p><b>CONCLUSION</b>Tau transgenic mice (F10) can still inherit their parents' biologiccal characters, and develop learning and memory dysfunction awnodh san obvious decrease in nitric oxide level of whole brain in the 7-month old mice, suggesting a decrease of nitric oxide level of whole brain would be involved in the mechanism of learning and memory dysfunction in these transgenic mice.</p>
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Animals , Humans , Mice , Acetylcholine , Metabolism , Acetylcholinesterase , Metabolism , Brain , Choline O-Acetyltransferase , Metabolism , Membrane Proteins , Genetics , Memory Disorders , Genetics , Mice, Transgenic , Mutation , Nitric Oxide , MetabolismABSTRACT
Background It is thought in recently that photodynamic therapy (PDT) is an effective treatment method for chronic central serous chorioretinopathy (CSC), but the dosage of verteporfin and its long-term efficacy and complications is rarely elucidated ever before.Objective This study was to observe the long-term efficacy and safety of 60％ dose verteporfin PDT for chronic CSC.Methods This is a retrospective study and a self-controlled design was used.The clinical data of 25 eyes of 21 chronic CSC patients who received 60％-dose verteporfin PDT in Henan Eye Institute from January 2009 to May 2010 were reviewed, with the male 18 (85.71％) and female 3 (14.29％) as well as monocular CSC 17 patients and binocular CSC 4 patients.The average ages of the patients were (43±5) years.Fundus fluorescein angiography (FFA) , indocyanine green angiography (ICGA), optical coherence tomography(OCT) and best corrected visual acuity (BCVA) were examined in all the patients before and after treatment.PDT with the 60％-dose verteporfin (3.6 mg/m2) was carried out on the CSC eyes.The treated eyes were examined 2 weeks, 1 month and 3 months after PDT.The BCVA,subfoveal choroid thickness,FFA and ICGA findings before and after PDT were compared.The following-up duration was 5 years or more.Results The BCVA before and 3 months after PDT were 0.5 ±0.1 and 0.9±0.2, respectively, with a statistically significant difference between them (t =19.17,P =0.00).The subfoveal choroidal thickness value 3 months after PDT was (326.56±39.47) μm,which was significantly reduced in comparison with (486.24 ±47.53) μm before PDT (t =25.17, P =0.00).FFA and ICGA showed that the leakage of fluorescein (hyperfluorescence) was disappeared in all the treated eyes.No systemic or local adverse effects and recurrence were observed during the follow-up period.Conclusions On the basis of the results of this study and available information,60％-dose verteporfin PDT seems to have a better long-term efficacy and safety than full-dose verteporfin in treating chronic CSC.
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<p><b>OBJECTIVE</b>To investigate the effect of hypobaric hypoxia (HH)on the cognitive function of mice and the phosphorylation of tau protein in mice brain.</p><p><b>METHODS</b>Forty male mice were randomly divided into 4 groups (n = 10): static control (control) group, 8 hours (8 h) group, 7 days(7 d) group and 28 days(28 d) group, which were exposed to simulated HH equivalent to 5 500 m in an animal decompression chamber for 0 hour, 8 hours, 7 days and 28 days, respectively. Cognitive performances were examined by open field and passive avoidance test, tan phosphorylation was assayed by Western blot.</p><p><b>RESULTS</b>In open field test,the group exposed in hypobaric hypoxia for 28 d showed lower mean velocity (P < 0.05), time in central zone (P < 0.05) was longer than control group. In passive avoidance test 28 d group presented worse performance in both latency time and number of mistakes (P < 0.05) compared with control group. Western blot showed that phosphorylated tau was increased significantly following exposure to HH for 7 d in cortex and 28 d in hippocampus (P < 0.05).</p><p><b>CONCLUSION</b>Tau hyperphosphorylation in brain of mice may play a role in chronic HH-induced cognitive function impairment.</p>
Subject(s)
Animals , Male , Mice , Cerebral Cortex , Metabolism , Disease Models, Animal , Hippocampus , Metabolism , Hypoxia , Metabolism , Maze Learning , Physiology , Memory , Physiology , Phosphorylation , tau Proteins , MetabolismABSTRACT
Objective To investigate the hepatitis B virus (HBV) infection status quo among childbearing age women to provide the theoretical basis for adopting the effective intervention ,treatment and blocking measures in pre-pregnant women .Methods The enzyme-linked immunosorbent assay (ELISA) and the time-resolved fluorometric immunoassay were adopted to detect HbsAg in 68 682 children bearing age women ,including outpatient and hospitalized patients ,in this hospital from February 2012 to September 2013 .The detect results were analyzed according to the age ,place of residence and medical departments .Results Among 68 682 children bearing age women ,3 260 cases were found positive for HbsAg with the HBsAg positive rate of 4 .70% .The statistically significant differences in the HBV infection positive rate existed among different age groups ,the positive rates in the age groups of 16- <20 years ,20 - < 25 years ,25 - < 30 years ,30 - < 35 years ,35 - < 40 years ,40 - < 45 years and 45 - < 50 years were 3 .83% ,4 .89% ,4 .96% ,4 .91% ,4 .18% ,4 .40% and 4 .18% respectively ,the differences among them had statistical significance (χ2 =15 .76 ,P=0 .015) ,in which the age 25- <30 years group had the highest HBV infection rate .At the same time the signifi-cant differences in the geographical distribution and medical departments existed .The HBV infection positive rate in rural area was significantly higher than that in city with statistical difference (χ2 =27 .47 ,P<0 .05) ,and the HBV infection positive rate in the outpatient departments was higher than that in the inpatient departments with statistical difference (χ2 =46 .88 ,P<0 .05) .Conclu-sion Children bearing women at 20-34 years old show the highest HBV infection rate .The HBV screening and the works of pre-vention and treatment during this age period should be strengthened ,which conduces to discover the HBV infection positive patients in time and prevent the mother-to-fetus transmission .
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Objective To investigate the morbidity of soldiers stationed on an island ,analyze the relationship betweendiseases and the environment,and to provide an effective method for disease prevention and treatment .Method Data ofoutpatients from troops stationed on an island between September 2012 and August 2013 were statistically investigatedaccording to the classification of diseases.The cause of disease was analyzed .In addition,90 soldiers stationed on an islandwere randomly selected to investigate their response to heat and humidity and parts susceptible to skin diseases viaquestionnaires and talks.Results A total of 789 cases of disease were identified,including 226 cases of upper respiratorytract infection,118 cases of orthopedic-related diseases,90 cases of traumatic diseases,88 cases of oral diseases,74 casesof digestive system diseases,62 cases of skin diseases,53 cases of ENT diseases,41 cases of urinary tract infection,and37 cases of anorectal diseases.The top five responses to heat and humidity were sweating,thirst,body fatigue,dizzinessand profuse sweating.The most susceptible part to skin diseases was the feet ,followed by the cheek,crotch,back andneck.Conclusion The epidemiological characteristics of diseases on this island are significant,and medical supportshould be focused on improving the overall level of hospital treatment.
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Objective To investigate the physical fitness status of stationed on an island and the influence factors of military operational ability in order to take effective intervention measures to cope with the problems encountered by the garrison force.Methods The outdoor environment heat intensity of the island was monitored and evaluated by thermal environment monitors.The physical fitness of thirty-eight soldiers randomly selected was evaluated by measuring VO 2max. Ninety soldiers stationed on this island were selected by random sampling , for whom questionnaires and interviews were designed, involving the influence factors of military operational ability .Results This island was a typical humid-hot environment, with high temperature , high humidity and high radiation .The physical fitness of soldiers declined obviously because of the typical humid-hot environment.The top five working environmental factors were high temperature , humidity, solar radiation, wind and seasonal drying .The top five personal factors were physical fatigue , overtraining, injury, mental fatigue and lack of sleep .Conclusion Considering the obviously decreased physical fitness of soldiers caused by the typical humid-hot environment , how to assess military combat effectiveness and how to determine whether the level of combat effectiveness can meet the actual needs of tropical combat , requires scientific indicators and criteria of evaluation .
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<p><b>OBJECTIVE</b>To observe the correlation between the decline of cognitive function and the level of plasma homocysteine in patients with Alzheimer's disease (AD).</p><p><b>METHODS</b>Thirty six AD patients were selected from hospitals in Tianjin. The enrolled patients were in accord with the diagnosis criteria. Thirty two control subjects were corresponding patients without AD in the period. Blood samples were extracted from each subject to determine the levels of homocysteine (Hcy) and folate. Cognitive status was evaluated by the mini- mental state examination (MMSE) and clinical dementia rating scale (CDR).</p><p><b>RESULTS</b>The mean value of serum Hcy concentration [(17.51 +/- 5.62) micromol/L] of AD group was higher than that of control group [(12.38 +/- 4.25)micromol/L]. The serum [(5.17 +/- 1.76) microg/L] and diet folate [(206.94 +/- 44.51) microg/d] concentration of AD group were lower than those of control group [(7.92 +/- 2.22) microg/L, (259.74 +/- 41.92) microg/ d]. The incidence of hyperhomocysteinemia in AD group (64%) was higher than that in control group (22%). A significant relation between Hcy concentrations and the CDR was observed. With the increase of Hcy concentrations the CDR raised, and with the increase of Hcy concentrations the MMSE decreased.</p><p><b>CONCLUSION</b>Hyperhomocysteinemia is one of the risk factors inducing the onset of AD. There is a significant negative correlation between Hcy levels and cognitive levels in AD group. Folate deficiency is an important reason to cause elevated Hcy levels in AD.</p>
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Humans , Alzheimer Disease , Blood , Case-Control Studies , Folic Acid , Blood , Homocysteine , Blood , Hyperhomocysteinemia , BloodABSTRACT
AIM@#To study the chemical constituents and bioactivity of the seeds of Crataegus pinnatifida.@*METHODS@#The chemical constituents were isolated and purified by macroporous adsorptive resin D101, silica gel, and ODS column chromatography, and preparative HPLC. Their structures were elucidated on the basis of spectroscopic methods. In addition, the cytotoxic activities of compounds 1-4 were investigated on OPM2 and RPMI-8226 cells.@*RESULTS@#Four compounds were obtained and their structures were identified as (7S, 8S)-4-[2-hydroxy-2-(4-hydroxy-3-methoxyphenyl)-1-(hydroxymethyl)ethoxy]-3, 5-dimethoxybenzaldehyde (1), (+)-balanophonin (2), erythro-guaiacylglycerol-β-coniferyl aldehyde ether (3), buddlenol A (4).@*CONCLUSION@#Compound 1 is a novel norlignan, while compounds 1-4 exhibited marginal inhibition on the proliferation of OPM2 and RPMI-8226 cells.
Subject(s)
Humans , Cell Line , Cell Proliferation , Crataegus , Chemistry , Molecular Structure , Nerve Tissue Proteins , Chemistry , Toxicity , Plant Extracts , Chemistry , Toxicity , Seeds , ChemistryABSTRACT
Background Researches have demonstrated that ocular hypertension induces the ischemia-reperfusion of retina and further leads to the degeneration of retinal ganglion cells,but its mechanism is beyond understanding.Objective The present study aims to observe the effects of static pressure on the morphology,proliferation activity and viability of cultured retinal microvascular endothelial cells (RMECs) and evaluate the expression of ET-1 and NO in these cells under variant static pressure.Methods RMECs were isolated from 30 healthy Wistar rats and cultured using explant culture method by Ⅷ factor antibody and PECAM-1 antibody.The static pressure of 1.33kPa,2.67kPa,5.33kPa and 10.67kPa was used in culture bottle respectively.The RMECs without static pressure were used as normal control group.The morphology of RMECs under the different static pressure was observed by inverted phase contrast microscopy,and the number of RMECs was counted using the counting plate.Cellular viability was studied by trypan blue staining.The changes of ET-1 and NO_2~-/NO_3~-,two metabolic products of NO,in the medium were detected by radioimmunoassay and Griess's nitrate reductase method.The expression of ET-1,eNOS and iNOS mRNA in RMECs was analyzed by semi-quantitative RT-PCR 24 hours after treatment of variant static pressure.Results Cultured RMECs sticked well at 24 hours and reached to confluence at 48 hours and showed the red fluorescence for Ⅷ factor antibody and PECAM-1 antibody.Enlargement of nuclei,extenders of cell bodies and suspension of RMECs in medium were observed.The number of RMECs was gradually increased.The cell viability was reduced with the raise of static pressure among these four groups(F=12.205,P＜0.01;F=11.180,P＜0.01).The static pressure increased the content of ET-1 released by RMECs in 2.67kPa,5.33kPa and 10.67kPa of static pressure groups,and concentrations of NO_2~-/NO_3~- in the medium showed a significant increase in 5.33kPa and 10.67kPa of static pressure groups compared with normal and 1.33kPa of static pressure groups(P＜0.01).The expressions of ET-1 mRNA,eNOS mRNA and iNOS mRNA were considerably enhanced in 5.33kPa and 10.67kPa of static pressure groups compared with normal control group(P＜0.01).Conclusion Raised static pressure causes the alteration of RMGCs structure and morphology.Static pressure could upregulate the expressions of ET-1,eNOS and iNOS mRNA in RMECs and increase the release of ET-1 and NO.This pathway might be one of pathologic mechanisms of retinal injury induced by high intraocular pressure.
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<p><b>OBJECTIVE</b>To investigate the material foundation of the fusion of bcr and abl genes, and to explore the pathogenesis of chronic myeloid leukemia.</p><p><b>METHODS</b>By FISH combined with laser confocal scanning microscopy, the three-dimension (3D) distribution of bcr and abl genes in the interphase nuclei of normal and irradiated IM-9 cells was studied in each cell cycle phases.</p><p><b>RESULTS</b>abl and bcr genes distributed non-randomly in the interphase nuclei of IM-9 cells. abl gene preferably located at the outer layer and bcr near the core of the nucleus. The two genes were drawn near each other most in G(0) phase. The relative distance between the homologous genes was greater at proliferation phase than at quiescence phase. After irradiation, the relative distances from the two genes to the core and between the two genes were shortened, with the shortest distance between the two genes in S phase.</p><p><b>CONCLUSION</b>Irradiation could change the 3D-distribution of abl and bcr genes in the interphase nuclei of IM-9 cell and accelerate them to draw near each other.</p>