Loss of GRB2 associated binding protein 1 in arteriosclerosis obliterans promotes host autophagy

Background:Arteriosclerosis obliterans (ASO) is a major cause of adult limb loss worldwide. Autophagy of vascular endothelial cell (VEC) contributes to the ASO progression. However, the molecular mechanism that controls VEC autophagy remains unclear. In this study, we aimed to explore the role of the GRB2 associated binding protein 1 (GAB1) in regulating VEC autophagy.Methods:In vivo and in vitro studies were applied to determine the loss of adapt protein GAB1 in association with ASO progression. Histological GAB1 expression was measured in sclerotic vascular intima and normal vascular intima. Gain- and loss-of-function of GAB1 were applied in VEC to determine the effect and potential downstream signaling of GAB1.Results:The autophagy repressor p62 was significantly downregulated in ASO intima as compared to that in healthy donor (0.80 vs. 0.20, t = 6.43, P < 0.05). The expression level of GAB1 mRNA (1.00 vs. 0.24, t = 7.41, P < 0.05) and protein (0.72 vs. 0.21, t = 5.97, P < 0.05) was significantly decreased in ASO group as compared with the control group. Loss of GAB1 led to a remarkable decrease in LC3II (1.19 vs. 0.68, t = 5.99, P < 0.05), whereas overexpression of GAB1 significantly led to a decrease in LC3II level (0.41 vs. 0.93, t = 7.12, P < 0.05). Phosphorylation levels of JNK and p38 were significantly associated with gain- and loss-of-function of GAB1 protein.Conclusion:Loss of GAB1 promotes VEC autophagy which is associated with ASO. GAB1 and its downstream signaling might be potential therapeutic targets for ASO treatment.     

中国医改之服务体系思考-关于“人”的思考体系之老年护理

医改内容很多，涉及方方面面，本文仅就老年护理这一改革的重点领域论述我国改革的一些成效，总结国、内外的有关经验，分析我国当前的现状并提出一些建议，为下一步全面推进医药卫生体制改革作参考。     

本地与外地高校大学生情绪稳定性

目的研究本地高校与外地高校大学生情绪稳定性的差异。方法以河北联合大学和齐齐哈尔大学共419名本科生为研究对象,采用艾森克人格问卷成人版(EPQ)进行测评,并对被试量表得分进行对比分析。结果①本地与外地男大学生在E(内外向)、P(精神质)方面均无显著性差异,在N(神经质)方面存在显著性差异(χ2=4.746,P<0.05);②本地与外地女大学生在E(内外向)、P(精神质)和N(神经质)方面均无显著性差异,但总体偏外向。结论本地男大学生情绪具有不稳定性,易出现强烈的情绪波动;外地男大学生情绪具有稳定性,善于自我控制。     

民间验方生存发展困境与保护策略

目的：探讨我国民间验方开发利用过程中存在的问题及其生存发展的困境，为合理获取和传承民间验方提出相关的建议和对策。方法：比较现行法律法规在保护民间验方上存在的不足与缺陷，借鉴泰国保护传统医药知识的做法和经验。结果：针对我国民间验方的实际情况，建议完善相关法律法规，建立民间验方保护名录。结论：建立民间验方保护名录，实行保密登记制度，可以对提出申请的民间验方中核心的信息进行保护，促进传统医药知识的传承与发展。     

Effect of different modeling time on intestinal bacteria in letrozole induced PCOS rats

Objective: The differences of ovarian morphology, reproductive hormones, glucose and lipid metabolism and intestinal bacteria in rats with polycystic ovary syndrome (PCOS) induced by triazole were compared. Method: Eighteen 21 SPF female SD rats were randomly divided into group A (3-week group), group B (5-week group) and group D (control group) by random number table.Group A received letrozole + CMC-Na mixture by gavage in the first 3 weeks and CMC-Na solution by gavage in the last 2 weeks, group B received letrozole + CMC-Na mixture by gavage for 5 weeks, and group D received CMC-Na solution by gavage for 5 weeks, and all three groups of rats were fed with normal diet.At the end of gavage, the body weight of rats in each group was observed, the histological changes of ovaries were observed by hematoxylin-eosin (HE) staining, the serum levels of estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), total cholesterol (TC), triglyceride (TG), fasting blood glucose (Glu), fasting insulin (FINS) and lipopolysaccharide (LPS) were measured by enzyme-linked immunosorbent assay (ELISA), and the LH/FSH ratio and insulin resistance index (HOMA IR) were calculated; the intestinal bacteria of rats were detected by 16S rRNA technique. Result: 1. Comparison of ovary histomorphology: Under light microscope, multiple luteum and oocytes were observed in mature follicles in group D, and granulosa cells were orderly arranged and multilayered, without cystic dilated follicles. There were no mature follicles in the ovarian tissues of group A and GROUP B. The follicles were irregular in structure and more cystic dilated follicles were visible. The number of granular cells in some follicles decreased or even disappeared. 2. Comparison of sex hormone levels: compared with group D, T level in group B was significantly increased (P < 0.001), and T level in group A had an upward trend (P > 0.05); The LH/FSH levels in group A and B were significantly increased (P < 0.001; P < 0.001). Compared with group A, E2 in group B was significantly decreased (P < 0.05) and T was significantly increased (P < 0.01). 3. Comparison of glucose and lipid metabolism levels: Compared with group D, TC levels in groups A and B were significantly increased (P < 0.01; P < 0.01). Compared with group A, TG in group B was significantly increased (P < 0.05). There were no significant differences in Glu, FINS and HOMA-IR levels among all groups. 4. Comparison of LPS levels: Compared with group D, the serum LPS levels of rats in groups A and B were significantly increased (P < 0.001; P < 0.01). 5. Intestinal flora analysis and comparison: At the phylum level, compared with group D, the abundance of Firmicutes in group B increased (P < 0.01), Firmicutes in group A showed an upward trend (P > 0.05), and the abundance of Bacteroidetes in groups A and B decreased (P < 0.05). At the genus level, compared with group D, Lactobacillus in group B increased (P < 0.01). The results of LEfSe analysis showed that there were differences in the composition of various intestinal bacteria among the three groups (LDA > 3).Conclusion: The phenotype of PCOS rats was related to the length of modeling, and the phenotypic characteristics of PCOS in rats at 5 weeks of modeling were more typical than those in rats at 3 weeks of modeling; PCOS can cause changes in intestinal flora, and the changes in the structure of intestinal flora between groups are related to different modeling duration.     

Matrigel‐Based Sprouting Endothelial Cell Culture System from Mouse Corpus Cavernosum Is Potentially Useful for the Study of Endothelial and Erectile Dysfunction Related to High‐Glucose Exposure

IntroductionA proper cavernous endothelial cell culture system would be advantageous for the study of the pathophysiologic mechanisms involved in endothelial dysfunction and erectile dysfunction (ED).AimTo establish a nonenzymatic technique, which we termed the “Matrigel‐based sprouting endothelial cell culture system,” for the isolation of mouse cavernous endothelial cells (MCECs) and an in vitro model that mimics in vivo situation for diabetes‐induced ED.MethodsFor primary MCEC culture, mouse cavernous tissue was implanted into Matrigel and sprouting cells from the tissue were subcultivated. To establish an in vitro model for diabetes‐induced ED, the primary cultured MCECs were exposed to a normal‐glucose (5 mmoL) or a high‐glucose (30 mmoL) condition for 48 hours.Main Outcome MeasuresThe purity of isolated cells was determined by fluorescence‐activated cell sorting analysis. MCECs incubated under the normal‐ or the high‐glucose condition were used for Western blot, cyclic guanosine monophosphate (cGMP) quantification, and in vitro angiogenesis assay.ResultsWe could consistently isolate high‐purity MCECs (about 97%) with the Matrigel‐based sprouting endothelial cell culture system. MCECs were subcultured up to the fifth passage and no significant changes were noted in endothelial cell morphology or purity. The phosphorylation of Akt and eNOS and the cGMP concentration were significantly lower in MCECs exposed to high glucose than in those exposed to normal glucose. MCECs exposed to the normal‐glucose condition formed well‐organized capillary‐like structures, whereas derangements in tube formation were noted in MCECs exposed to high glucose. The protein expression of transforming growth factor‐β1 (TGF‐β1) and phospho‐Smad2 was significantly increased by exposure to high glucose.ConclusionThe Matrigel‐based sprouting endothelial cell culture system is a simple, technically feasible, and reproducible technique for isolating pure cavernous endothelial cells in mice. An in vitro model for diabetic ED will be a valuable tool for evaluating the angiogenic potential of novel endogenous or synthetic modulators. Yin GN, Ryu J‐K, Kwon M‐H, Shin SH, Jin HR, Song K‐M, Choi MJ, Kang D‐Y, Kim WJ, and Suh J‐K. Matrigel‐based sprouting endothelial cell culture system from mouse corpus cavernosum is potentially useful for the study of endothelial and erectile dysfunction related to high‐glucose exposure. J Sex Med 2012;9:1777–1789.