目的:研究缺氧预处理对老年大鼠子宫及双附件切除术后疲劳是否有改善作用,并通过对比观察超氧化物歧化酶及丙二醛水平的变化,初步探讨缺氧预处理的作用机制。方法:将老年大鼠分为空白对照组、对照组、缺氧预处理三组。空白对照组为假手术组,对照组为子宫及双附件切除术组, 缺氧预处理组为缺氧预处理加子宫及双附件切除术组。对比观察缺氧预处理对大鼠体力活动及血清超氧化物歧化酶和丙二醛水平的影响。结果:空白对照组、对照组、缺氧预处理三组大鼠悬尾不动时间分别为:(21±3)s,(83±10)s,(44±5)s,各组间比较Plt;0.05。三组SOD活性分别为:(131.23±5.31)U/L,(36.12±9.68)U/L,(73.01±9.82)U/L,各组间比较Plt;0.05。三组MDA水平分别为:(9.78±1.26)μmol/L,(29.87±3.13)μmol/L,(15.98±2.21)μmol/L,各组间比较Plt;0.05。结论:缺氧预处理可提高老年大鼠的抗氧化能力,对老年大鼠子宫及双附件切除术后疲劳综合征有明显的改善作用。
Objective To study the protective effects of bone marrow mesenchymal stem cells (BMSCs) of rhesus monkeys on porcine islets from hypoxia/reoxygenation (H/R)-induced injury. Methods BMSCs were isolated and cultured from the marrow of 5 adult rhesus monkeys (weighing, 6-10 kg) by adherent monocytes. Islets were isolated and purified from the pancreas of 5 neonatal porcine (3-5 days old) by collagenase V digestion method, and were cultured with or without BMSCs, and exposed to hypoxia (1%O2) for 12 hours and reoxygenation for 24 or 48 hours, respectively. The experiment was divided into 4 groups: normal islet group (group A), normal islet + BMSCs group (Group B), H/R islet group (group C), and H/R islet + BMSCs group (group D). The survival rate of islets was calculated by fluorescein diacetate/propidium iodide (PI) staining. The viability of the islet cells was detected by cell counting kit 8. Apoptotic rate of islet cells was tested using Annexin V-FITC/PI labeling and flow cytometry. The stimulation index (SI) of islet function was analyzed by glucose-stimulated insulin secretion assay. Results The islet cell cluster of group C was more dispersed than that of groups A and B, and group C had more death cells; and the islet cell cluster of group D was more complete and the survival rate was higher than those of group C. The survival rate of islet was 90.2% ± 9.1%, 88.3% ± 5.9%, 52.3% ± 12.1%, and 71.4% ± 11.5% in groups A, B, C, and D respectively, it was significantly lower in groups C and D than in groups A and B (P lt; 0.05), but it was significantly higher in group D than in group C (P lt; 0.05). After coculture of BMSCs and islet at the ratio of 1 ∶ 10 and 1 ∶ 20 in group D, the viability of islet cells was significantly higher than that in group C (P lt; 0.05). The apoptotic rate was 27.1% ± 3.2%, 24.0% ± 1.0%, 64.3% ± 1.8%, and 46.2% ± 1.4% in groups A, B, C, and D respectively, it was significantly higher in groups C and D than that in groups A and B (P lt; 0.05), but it was significantly lower in group D than in group C (P lt; 0.05). There was no significant difference in SI between groups A and B at each time point (P gt; 0.05), but it was significantly lower in group C than in groups A and B (P lt; 0.05); and it was significantly higher in group D than in group C at 24 and 72 hours (P lt; 0.05). Conclusion BMSCs of rhesus monkeys can protect islet vitality and function from H/R-induced injury.
ObjectiveTo investigate the renal impairment and the risk factors of renal impairment in patients with OSA. MethodsData from patients who underwent polysomnography (PSG) in our department from July 2022 to January 2023 were collected, totaling 178 cases. Based on the results of the polysomnography, the patients were divided into an OSA group (145 cases) and a non-OSA group (33 cases). According to the severity of the condition, the OSA group was further divided into mild OSA (21 cases), moderate OSA (28 cases), and severe OSA (96 cases). The Pearson correlation analysis was further conducted to analyze the relationships between serum urea nitrogen (BUN), serum cystatin C (Cys-C) concentrations, and estimated Glomerular Filtration Rate (eGFR) with various risk factors that may influence renal impairment. Moreover, multiple linear regression analysis was used to identify the risk factors affecting BUN, Cys-C, and eGFR. ResultsWhen comparing the two groups, there were statistically significant differences in age, weight, BMI, neck circumference, waist circumference, eGFR、Cys-C、BUN, LSaO2, CT90% (all P<0.05). Univariate analysis of variance was used to compare differences in BUN, Serum creatinine (SCr), Cys-C, and eGFR among patients with mild, moderate, and severe OSA, indicating that differences in eGFR and Cys-C among OSA patients of varying severities were statistically significant. Further analysis with Pearson correlation was conducted to explore the associations between eGFR, BUN, and Cys-C with potential risk factors that may affect renal function. Subsequently, multiple linear regression was utilized, taking these three indices as dependent variables to evaluate risk factors potentially influencing renal dysfunction. The results demonstrated that eGFR was negatively correlated with age, BMI, and CT90% (β=−0.95, P<0.001; β=−1.36, P=0.01; β=−32.64, P<0.001); BUN was positively correlated with CT90% (β=0.22, P=0.01); Cys-C was positively correlated with CT90% (β=0.58, P<0.001. Conclusion Chronic intermittent hypoxia, age, and obesity are risk factors for renal dysfunction in patients with OSA.
Objective To investigate the effect of ginkgolide B (GB) on cysteinyl aspartate specific proteinase-3 (Caspase-3)/chromosome 10 deletion phosphatase-tension protein homologue (PTEN)/protein kinase B (Akt) pathway and cell proliferation and apoptosis in hypoxia/reoxygenation cardiomyocytes. Methods H9C2 cells were cultured in vitro. A control group was cultured in serum-free DMEM high glucose medium at 37°C and 5% CO2 for 28 hours. The remaining groups were prepared with hypoxia/reoxygenation models. A GB low-dose group and a GB high-dose group were treated with GB pretreatment with final concentration of 50 μmol/L and 200 μmol/L respectively at 1 h before hypoxia/reoxygenation. A carvedilol group was treated with carvedilol of a final concentration of 10 μmol/L at 1 h before hypoxia/reoxygenation. The proliferation and apoptosis of H9C2 cells were detected, and the levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), reactive oxygen species (ROS), PTEN, Akt, phosphorylated Akt (p-Akt) and Caspase-3 in H9C2 cells were also detected. Results Compared with the control group, the proliferation rate of H9C2 cell, and the levels of PTEN, Akt and p-Akt in other groups decreased, and the apoptosis rate, and the levels of LDH, MDA, ROS and Caspase-3 increased (P<0.05). Compared with the hypoxia/reoxygenation group, the proliferation rate of H9C2 cell, and the levels of PTEN, Akt and p-Akt in all GB dose groups and the carvedilol group increased; the apoptosis rate, and the levels of LDH, MDA, ROS and Caspase-3 decreased, and the effect of GB was in a dose dependent manner; however, the effect of GB was not as strong as carvedilol (P<0.05). Conclusion GB can inhibit H9C2 cell apoptosis and promote H9C2 cell proliferation by activating Caspase-3/PTEN/Akt pathway.
Objective To observe the influence of the expression of CD18 on the neutrophile and the leukocyte adhesion to retinal vascular endothelium by hypoxia-inducible factor-1 alpha (HIF-1alpha;) in early diabetic retinopathy rats. Methods Male Sprague-Dawley rats received intraperitoneal injection of streptozotocin to induce diabetes model. 18 diabetic rats were divided into 3 groups randomly after 2 months of diabetes induction, including diabetic group (group B), HIF-1alpha; anti-sense oligonucleotides (ASODN) injection group (group C) and HIF-1alpha; sense oligonucleotides (SODN) injection group (group D), the age and weigh matched health rats were chosen as control group (group A), with 6 rats in each group. Then group A and B rats received 5% glucose solution caudalis veins injection, group C and group D rats received HIF-1alpha; ASODN and HIF-1alpha; SODN caudalis veins injection, respectively(025 mg/kg).The level of CD18 on the neutrophil isolated from the peripheral blood was measured by flow cytometry. Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Results The percentage of CD18 positive neutrophil cell was（44.93plusmn;3.60）% in group B，（18.66plusmn;1.52）% in group A，（31.66plusmn;4.72）% in group C，（51.00plusmn;5.66）% in group D. Compared with each other groups，the differences are statistically significant （F=42.46, Plt;0.001）. The number of positive staining cells of retinal leukocyte was （46.16plusmn;10.68）in group A，（133.83plusmn;20.43）in group B,（99.83plusmn;9.28）in group C，（121.33plusmn;10.23） in group C. Compared group B with group C，the number of positive staining cells raised about 2.89 times；compared group B with group C and D，the differences are statistically significant （P=0.12，95% confidence interval -3.69~28.69）. Conclusions In vivo, HIF-1alpha; can decreased the expression of CD18 on neutrophils from diabetic ratsprime; peripheral blood and the collection of retinal leukostasis in the diabetic animals. HIF-1alpha; may serve as a therapeutic target for the treatment and/or prevention of early diabetic retinopathy. （Chin J Ocul Fundus Dis，2008，24：268-271）
ObjectiveTo objectively evaluate the effect and safety of naloxone for the treatment of moderate and severe neonatal hypoxia-ischemic encephalopathy (HIE). MethodsResearch articles published from inception to June 2015 on Cochrane library, PubMed, Web of science, Chinese Science and Technology Journal Full-text Database, Digital Full-text Journal Database and Chinese Journal Full-text Database were searched, which were relevant to naloxone in the treatment of moderate and severe neonatal HIE. And two authors extracted information via standardized data extraction form and assessed the quality of included studies independently. RevMan 5.2 software was used for Meta-analysis. ResultsAt last, 20 randomized controlled trials (involving 1 519 neonates; 783 in the treatment group and 736 in the control group) were included. The results of meta-analysis showed that the effect of naloxone for moderate and severe HIE was significantly superior to the control group[OR=5.07, 95%CI (3.61, 7.12), P < 0.000 01]. The neurobehavioral scores in neonates treated by naloxone after 5, 7, 10, and 14 days were higher than those in the control group[WMD=6.61 points, 95%CI (5.70, 7.51) points, P < 0.000 01; WMD=4.27 points, 95%CI (2.63, 5.91) points, P < 0.000 01; WMD=2.40 points, 95%CI (1.47, 3.34) points, P < 0.000 01; WMD=2.58 points, 95%CI (1.00, 4.16) points, P=0.001], respectively; while the neurobehavioral scores after 3-day treatment between the two groups had no statistically difference[WMD=0.00 points, 95%CI(-0.76, 0.77) points, P=0.99]. What's more, the disappeared time of clinical symptoms and signs (breathing improvement time, recovery time, convulsions disappearance time, and signs disappearance time) in naloxone group was superior to the control groups[WMD=-3.78 hours, 95%CI (-6.93, -0.64) hours, P=0.02; WMD=-9.66 hours, 95%CI (-14.25, -5.06) hours, P < 0.001; WMD=-2.81 hours, 95%CI (-5.28, -0.35) hours, P=0.03; WMD=-1.02 days, 95%CI (-1.84, -0.20) days, P=0.01], respectively. ConclusionsNaloxone has certain therapeutic on moderate and severe HIE. Further high-quality randomized controlled trials should be carried out to provide more reliable evidence.
ObjectiveTo investigate the effect of LOC103693069 on hypoxic apoptosis of bone marrow mesenchymal stem cells (BMSCs). Methods BMSCs from 1-week-old Sprague Dawley rat bone marrow were isolated, cultured, and passaged by the whole bone marrow adherent culture method. After identification of adipogenic, chondrogenic, and osteogenic differentiation, the 3rd generation cells were treated with hypoxia under 5%O2, 1%O2, and anaerobic conditions. After 48 hours, the cell viability, apoptosis, and apoptosis-related proteins [hypoxia inducible factor 1α (HIF-1α), Caspase-3, B cell lymphoma/leukemia 2 (Bcl-2)] expressions were detected, and normal BMSCs were used as controls. Based on the research results, the concentration group with the most obvious apoptosis was selected and used for subsequent experiments. After 48 hours of hypoxia treatment, BMSCs were taken and analyzed by gene chip and real-time fluorescence quantitative PCR (qRT-PCR) to screen the most significantly down-regulated gene and construct their high-expression, low-expression, and negative control lentiviruses; BMSCs were transfected with the different lentiviruses, respectively. After qRT-PCR detection confirmed that the transfection was successful, the BMSCs were treated with hypoxia for 48 hours to observe the cell viability and the expressions of apoptosis-related proteins. ResultsAfter cell viability, apoptosis, and apoptosis-related proteins were detected, cell apoptosis was the most significant under anaerobic conditions after 48 hours. The above indicators were significantly different from other groups (P<0.05), and this group was used for treatment conditions for subsequent experiments. Gene chip analysis showed that after 48 hours of hypoxia treatment, AC125847.1, LOC102547753, AABR07017208.2, and LOC103693069 were significantly down-regulated in BMSCs, and the expressions of LOC103693069 was the most significant down-regulation detected by qRT-PCR (P<0.05). It was selected to construct lentivirus and transfect BMSCs. Afterwards, qRT-PCR detection showed the successful transfection into the cells. After hypoxia treatment, the apoptosis rate and the expressions of apoptosis-related proteins of BMSCs overexpressed by the gene were significantly reduced (P<0.05). Conclusion LOC103693069 can relieve the hypoxic apoptosis of BMSCs.
Objective To study the mechanisms and treatment of ischemia /reperfusion injury, expression of intracellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) were measured, the effect on suppression of ICAM-1 and VCAM-1 by the pyrrolidine dithiocarbamate (PDTC) were investigated. Methods Endothelial cells were divided into 3 groups, hypoxia group: endothelial cells were exposed in hypoxia condition, then returned to reoxygenation condition; the PDTC group: PDTC was added to the endothelial cells in the culture media before exposing to hypoxia condition; control group: endothelial cells underwent treatment. Confocal microscopy was used to detect expression of ICAM-1 and VCAM-1. Results ICAM-1 and VCAM-1 expression were low in endothelial cells of control group, and increased in hypoxia group . ICAM-1 and VCAM-1 expression of endothelial cells in PDTC group werelower than those in hypoxia group , but higher than those in control group. Conclusions It seems that hypoxia/ reoxygenation can activate the endothelial cells and increase the expression of cell adhesion molecules. PDTC can decrease the expression of ICAM-1 and VCAM-1. PDTC may prove benificial in the treatment of ischemia /reperfusion injury.