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.     

神经刺激仪定位在老年桡骨骨折手术患者行肌间沟臂丛神经阻滞麻醉中的应用价值

目的 探讨神经刺激仪定位用于老年桡骨骨折患者行肌间沟臂丛神经阻滞麻醉效果。 方法 选择行肌间沟臂丛神经阻滞老年桡骨骨折患者100例,随机分为S组(50例)和P组(50例),分别在神经刺激仪和传统异感法定位下予罗哌卡因行神经阻滞,比较两组患者麻醉效果及满意度。 结果 S组操作,臂丛感觉神经阻滞起效、完善及运动神经阻滞起效时间均短于P组,差异有统计学意义(P<0.05);S组麻醉满意度优于P组,差异有统计学意义(P<0.05)。结论 神经刺激仪定位应用于老年患者行肌间沟臂丛神经阻滞,安全有效。     

血管内超声指导药物洗脱支架置入对合并慢性肾脏病患者长期预后的影响:ULTIMATE CKD亚组分析

目的评估血管内超声(IVUS)指导药物洗脱支架(DES)置入对慢性肾脏病(CKD)患者长期临床结局的影响。方法研究对象来源于ULTIMATE研究,该研究是一项前瞻性、多中心、随机对照研究,从2014年8月到2017年5月在国内8家中心入选1448例置入DES的冠心病患者,按1∶1的比例随机分为两组(接受IVUS或冠状动脉造影指导支架置入)。本研究选取ULTIMATE研究中有基线血肌酐值的患者1443例,分为CKD组和非CKD组。CKD的定义为Cockcroft-Gault(CG)公式得出的估算的肾小球滤过率(eGFR)<60 ml·min^-1·1.73 m^-2至少3个月。主要终点是术后3年靶血管失败(TVF),包含心原性死亡、靶血管心肌梗死和临床症状驱动的靶血管血运重建。采用Kaplan-Meier法进行生存分析,通过log-rank检验比较各组的终点事件发生情况,并采用Cox风险比例模型计算HR及其95%CI,同时检验交互作用。采用多因素Cox回归分析TVF的独立影响因素。结果本研究共入选1443例置入DES的冠心病患者,其中CKD组349例(24.2%),非CKD组1094例。CKD组中采用IVUS指导支架置入者180例,造影指导支架置入者169例;非CKD组中采用IVUS指导支架置入者543例,造影指导支架置入者551例。3年临床随访率为98.3%(1418/1443)。术后3年时,CKD组的TVF发生率为12.0%(42/349),高于非CKD组的7.4%(81/1094)(P=0.01),其差异主要来源于CKD组更高的心原性死亡率[4.6%(16/349)比1.5%(16/1094),P<0.001]。CKD组中IVUS指导下置入支架者的TVF发生率低于造影指导下置入支架者[8.3%(15/180)比16.0%(27/169),P=0.03]。而非CKD组共81例发生TVF,其中IVUS指导下置入支架者的TVF发生率与造影指导下置入支架者差异无统计学意义[5.9%(32/543)比8.9%(49/551),P=0.06],不存在交互作用(P=0.47)。多因素Cox回归分析结果显示,IVUS指导(HR=0.56,95%CI 0.39~0.81,P=0.002)、CKD(HR=1.83,95%CI 1.17~2.87,P=0.010)和支架长度(每增加10 mm)(HR=1.11,95%CI 1.04~1.19,P=0.002)是置入DES术后3年发生TVF的独立影响因素。结论对于置入DES的冠心病患者,合并CKD者术后3年的TVF发生风险高于非CKD患者;相比于冠状动脉造影,IVUS指导DES置入可以降低CKD患者的TVF风险。     

A new thrombolytic agent,monteplase, is independent of the plasminogen activator inhibitor in patients with acute myocardial infarction: initial results of the COmbining Monteplase with Angioplasty (COMA) trial

Background Both thrombolytic therapy and coronary angioplasty have been inconsistent together for primary acute myocardial infarction (AMI) therapy, because conventional thrombolytic agents accelerate plasminogen activator inhibitor-1 (PAI-1) activity. However, combining newly developed mutant tissue-type plasminogen activators with coronary angioplasty should be reconsidered. Methods This study was designed to investigate clinical usefulness of such an agent, monteplase, for treatment of AMI in light of PAI-1 kinetics. One hundred fifty-four consecutive patients with AMI were randomly assigned to receive direct coronary angioplasty (Group I) or coronary angioplasty after pretreatment with intravenous monteplase (Group II). In 90 of these patients, total PAI-1 antigen levels were serially measured. Results Baseline PAI-1 levels at admission were higher in patients with occluded infarct-related arteries than in patients with patent arteries in Group I (39 ± 4 vs 20 ± 2 ng/mL, P < .01) and in Group II (36 ± 3 vs 27 ± 2 ng/mL, P < .05). In the high PAI-1 level subgroup (≥27 ng/mL, n = 53), Group II showed a higher patency rate than Group I (56 vs 18%, P < .01). Multiple logistic regression analysis indicated that patency could be predicted by the PAI-1 level in Group I (Wald χ²= 3.94, P = .02, odds ratio 0.924, 95% CI 0.855-0.999), but not in Group II. Serial change patterns in the PAI-1 level were identical in Group I and Group II. Conclusion Because monteplase can be used independently of PAI-1 kinetics, a combination of monteplase administration at a community hospital with prompt transfer to a tertiary center for coronary intervention may be a powerful strategy for AMI. (Am Heart J 2002;144:e5.)     

Genomic alterations as mediators of miRNA dysregulation in ovarian cancer

Ovarian cancer is the fifth most common cause of cancer death in women worldwide. Serous epithelial ovarian cancer (SEOC) is the most common and aggressive histological subtype. Widespread genomic alterations go hand‐in‐hand with aberrant DNA damage signaling and are a hallmark of high‐grade SEOC. MicroRNAs (miRNAs) are a class of small noncoding RNA molecules that are nonrandomly distributed in the genome. They are frequently located in chromosomal regions susceptible to copy number variation (CNV) associated with malignancy that can influence their expression. Widespread changes in miRNA expression have been reported in multiple cancer types including ovarian cancer. This review examines CNV and single nucleotide polymorphisms, two common types of genomic alterations that occur in ovarian cancer, in the context of their influence on the expression of miRNA and the ability of miRNA to bind to and regulate their target genes. This includes genes encoding proteins involved in DNA repair and the maintenance of genomic stability. Improved understanding of mechanisms of miRNA dysregulation and the role of miRNA in ovarian cancer will provide further insight into the pathogenesis and treatment of this disease. © 2014 Wiley Periodicals, Inc.