Ischemic injury of the liver in a porcine model of cardiac death assessed by in vivo microdialysis

This study aims to evaluate the ischemic injury of the liver in a porcine model of cardiac death assessed by in vivo microdialysis. A porcine model of cardiac death was established by the suffocation method. Metabolic indicators were monitored using the microdialysis technique during warm ischemia time (WIT) and cold ischemia time (CIT). Pathological changes in ischemic-injured livers were observed by haematoxylin–eosin staining. The predictive values of biochemical parameters regarding the liver donor were evaluated by receiver operating characteristic curve analysis. All statistical analyses were conducted using the SPSS 18.0 software (SPSS Inc, Chicago, Illinois, USA). The degree of warm ischemic injury of the livers increased with prolonged WIT. Serum glucose, glycerol, pyruvate, lactic acid levels and lactate-to-pyruvate (L/P) ratio increased gradually during WIT. Results from Pearson correlation analyses indicated that serum lactate level and L/P ratio were positively associated with the degree of warm ischemic injury of the livers. The degree of cold ischemic injury of the livers gradually increased after 12 h CIT. Serum glucose, lactic acid and L/P ratio achieved a peak after 6–8 h of CIT, but gradually decreased with prolonged CIT. The peak of glycerol occurred after 8 h of CIT, while no changes were found with prolonged CIT. Serum pyruvate level exhibited an increasing trend after 12 h CIT. Our results confirmed that serum glucose and lactate levels were negatively correlated with cold ischemic injury of the liver. However, serum glycerol and pyruvate levels showed positive correlations with cold ischemic injury of the liver. The liver donor was unavailable after 30 min WIT and 24 h CIT. The cut-off value of serum lactate level for warm ischemic injury of the livers was 2.374 with a sensitivity (Sen) of 90 % and specificity (Spe) of 95 %; while the L/P radio was 0.026 (Sen = 80 %, Spe = 83 %). In addition, the cut-off values of serum glucose, lactate, glycerol and pyruvate levels for cold ischemic injury of the livers were 0.339 (Sen = 100 %, Spe = 77 %), 1.172 (Sen = 100 %, Spe = 61 %), 56.359 (Sen = 100 %, Spe = 65 %) and 0.020 (Sen = 100 %, Spe = 67 %), respectively. Our findings provide empirical evidences that serum glucose, lactate levels and L/P ratio may be good indicators for the degree of warm ischemic injury of the livers after cardiac death; while serum glucose, lactate, glycerol and pyruvate levels may be important in predicting cold ischemic injury.     

The Rho kinase inhibitor Y-27632 facilitates the differentiation of bone marrow mesenchymal stem cells

The selective in vitro expansion and differentiation of multipotent stem cells are critical steps in cell-based regenerative therapies, but technical challenges have limited cell yield and thus the success of these potential treatments. The Rho GTPases and downstream Rho kinases (Rho coiled-coil kinases or ROCKs) are central regulators of cytoskeletal dynamics during the cell cycle and thus help determine the balance between stem cells self-renewal, lineage commitment, and apoptosis. Here, we examined if suppression of ROCK signaling enhances the efficacy of bone marrow-derived mesenchymal stem cells (BMSCs) differentiation into neurons and neuroglial cells. BMSCs were cultured in epidermal growth factor (EGF, 10 µg/l) and basic fibroblastic growth factor (bFGF, 10 µg/l) in the presence or absence of the Rho kinase inhibitor Y-27632 (10 µM). The expression levels of neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) were detected by immunofluorescence and Western blotting. The average number of NSE-positive cells increased from 83.20 ± 8.677 (positive ratio 0.2140 ± 0.0119) to 109.20 ± 8.430 (positive ratio 0.3193 ± 0.0161) per visual field in the presence of Y-27632, while GFAP-positive cell number increased from 96.30 ± 8.486 (positive ratio 0.18 ± 0.0152) to 107.50 ± 8.683 (positive ratio 0.27 ± 0.0115) (P < 0.05 for both). Both NSE and GFAP protein expression levels were enhanced significantly by Y-27632 treatment (NSE: 0.74 ± 0.05 vs. 1.03 ± 0.06; GFAP: 0.64 ± 0.08 vs. 0.97 ± 0.05, both P < 0.01) as indicated by Western blots. The Rho kinase inhibitor Y-27632 concomitant with EGF and bFGF stimulation promotes BMSC differentiation into neural cells. Control of Rho kinase activity may enhance the efficiency of stem cell-based treatments for neurodegenerative diseases.     

B超引导下甲状腺肿块细针穿刺细胞学诊断的临床病理分析

目的：比较B超引导下细针吸取穿刺和传统细针吸取穿刺在甲状腺肿块性质确定中意义。方法：使用B超引导下细针吸取穿刺（UG-FNAB）和传统细针吸取穿刺技术（C-FNAB）对225例甲状腺肿块进行穿刺活检,然后进行涂片,染色,镜检,结合临床作出细胞学诊断,并与组织切片诊断对照,比较两种方法的确诊率。结果：在UG—FNAB检测病例中59.3％的结节位于甲状腺右叶,40．7％肿块位于左叶和峡部。87％的患者甲状腺机能正常,结节最长径平均为2．8±1．1cm,部分病例符合甲状腺恶性肿瘤的超声影像学改变。在本研究中诊断为甲状腺乳头状癌的病例细胞学和组织学特点均符合甲状腺癌形态学特点。在225例研究病例中,总确诊率为72．89％（164／225）,其中UG-FNAB组确诊率为90．58％（77／85）,C-FNAB组确诊率为62．14％（87／140）,两组确诊率比较有显著性差异（P〈0．05）。结论：本组研究数据显示比较C—FNAB,UG—FNAB在诊断甲状腺复杂性结节中具有更高的确诊率,特别是在C—FNAB方法不能得出明确诊断时。     

单纯治疗慢性前列腺炎对其合并早泄的影响

目的:探讨单纯治疗慢性前列腺炎对其合并早泄的影响。方法:选择372例前列腺炎继发早泄的患者进行单纯针对前列腺炎的治疗,评价前列腺炎的治疗效果(包括前列腺炎症状评分(NIH-CPSI)、前列腺液常规等)及早泄的治疗效果(包括患者性生活满意度评分、配偶性生活满意度评分及阴道内射精潜伏期等)。结果:慢性前列腺炎经综合治疗后,患者的NIH-CPSI评分及前列腺液白细胞计数均显著降低(P0.05)。前列腺炎治愈或好转后,大多数患者的早泄情况得到明显改善,患者性生活满意度、配偶性生活满意度均较治疗前显著提高,阴道内射精潜伏期亦较治疗前明显延长,差异均具有统计学意义(P0.05)。结论:单纯治疗慢性前列腺炎继可使大部分患者并发的早泄明显改善,而对少数前列腺炎好转后早泄症状改善不明显者,可联合应用SSRIs等药物治疗。     

UDP-glucose dehydrogenase modulates proteoglycan synthesis in articular chondrocytes: its possible involvement and regulation in osteoarthritis

Introduction

The objective of this study was to investigate the possible role of UDP-glucose dehydrogenase (UGDH) in osteoarthritis (OA) and uncover whether, furthermore how interleukin-1beta (IL-1β) affects UGDH gene expression.

Methods

UGDH specific siRNAs were applied to determine the role of UGDH in proteoglycan (PG) synthesis in human articular chondrocytes. Protein levels of UGDH and Sp1 in human and rat OA cartilage were detected. Then, human primary chondrocytes were treated with IL-1β to find out whether and how IL-1β could regulate the gene expression of UGDH and its trans-regulators, that is Sp1, Sp3 and c-Krox. Finally, p38 mitogen-activated protein kinase (MAPK) inhibitor SB203580 and stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) inhibitor SP600125 were used to pick out the pathway that mediated the IL-1β-modulated PGs synthesis and gene expression of UGDH, Sp1, Sp3 and c-Krox.

Results

UGDH specific siRNAs markedly inhibited UGDH mRNA and protein expression, and thus led to an obvious suppression of PGs synthesis in human articular chondrocytes. UGDH protein level in human and rat OA cartilage were much lower than the corresponding controls and negatively correlated to the degree of OA. Decrease in Sp1 protein level was also observed in human and rat OA cartilage respectively. Meanwhile, IL-1β suppressed UGDH gene expression in human articular chondrocytes in the late phase, which also modulated gene expression of Sp1, Sp3 and c-Krox and increased both Sp3/Sp1 and c-Krox/Sp1 ratio. Moreover, the inhibition of SAP/JNK and p38 MAPK pathways both resulted in an obvious attenuation of the IL-1β-induced suppression on the UGDH gene expression.

Conclusions

UGDH is essential in the PGs synthesis of articular chondrocytes, while the suppressed expression of UGDH might probably be involved in advanced OA, partly due to the modulation of p38 MAPK and SAP/JNK pathways and its trans-regulators by IL-1β.

Electronic supplementary material

The online version of this article (doi:10.1186/s13075-014-0484-2) contains supplementary material, which is available to authorized users.     

High-Glucose Inhibits Human Fibroblast Cell Migration in Wound Healing via Repression of bFGF-Regulating JNK Phosphorylation

One of the major symptoms of diabetes mellitus (DM) is delayed wound healing, which affects large populations of patients worldwide. However, the underlying mechanism behind this illness remains elusive. Skin wound healing requires a series of coordinated processes, including fibroblast cell proliferation and migration. Here, we simulate DM by application of high glucose (HG) in human foreskin primary fibroblast cells to analyze the molecular mechanism of DM effects on wound healing. The results indicate that HG, at a concentration of 30 mM, delay cell migration, but not cell proliferation. bFGF is known to promote cell migration that partially rescues HG effects on cell migration. Molecular and cell biology studies demonstrated that HG enhanced ROS production and repressed JNK phosphorylation, but did not affect Rac1 activity. JNK and Rac1 activation were known to be important for bFGF regulated cell migration. To further confirm DM effects on skin repair, a type 1 diabetic rat model was established, and we observed the efficacy of bFGF on both normal and diabetic rat skin repair. Furthermore, proteomic studies identified an increase of Annexin A2 protein nitration in HG-stressed fibroblasts and the nitration was protected by activation of bFGF signaling. Treatment with FGFR1 and JNK inhibitors delayed cell migration and increased Annexin A2 nitration levels, indicating that Annexin A2 nitration is modulated by bFGF signaling via activation of JNK. Together with these results, our data suggests that the HG-mediated delay of cell migration is linked to the inhibition of bFGF signaling, specifically through JNK suppression.     

模拟酸雨对朱砂叶螨寄主植物三月早茄生理生化的影响

以朱砂叶螨的寄主植物三月早茄为对象,研究了不同pH值和不同时间模拟酸雨胁迫下,茄子对酸雨的抵抗、适应能力和营养物质含量变化.结果表明,随酸雨酸度增强,茄叶的过氧化氢酶(CAT)活性、磷和可溶性蛋白含量呈单峰变化趋势;而过氧化物酶(POD)活性、可溶性糖含量呈单谷变化趋势;还原糖含量和超氧化物歧化酶(SOD)活性随酸度增加呈上升趋势;茄叶pH值呈下降趋势,含水量不受酸雨影响.POD对酸雨作用反应迟缓,而SOD、CAT较敏感,其他几项生理指标则对酸雨作用时间反应灵敏.高pH(4.0～5.6)的酸雨刺激茄叶的保护能力增强,促进茄叶的生长发育.由于茄叶内糖含量、磷及可溶性蛋白含量的改变,植物的适口性有所变化,可促进叶螨生长,而在强酸性酸雨(pH＜3.0)胁迫下,植物的生长受到抑制,营养物质含量降低,抑制朱砂叶螨的生长.     

烟草未授粉子房胚状体诱导的研究

对烟草（Ｎｉｃｏｔｉａｎａ ｔａｂａｃｕｍ Ｌ．）未授粉子房胚状体的诱导进行了研究,结果发现,胚状体是单细胞起源,起源于大孢子或卵细胞,接种发育早期的子房,胚状体起源于大孢子;接种发育晚期的子房,胚状体起源于卵细胞;接种发育中期的子种,胚状多起源于大孢子,少数起源于卵细胞,不同的基因型,蔗糖浓度及光照强度等对胚状体诱导率的影响亦有不同。     

满江红鱼腥藻遗传多样性的RAPD分析

共生在水生蕨类植物满江红(又称“红萍”:Azolla)叶腔中的固氮兰细菌曾被认为是一个种满江红鱼腥藻(Anabaena azollae Strasb.).但由于现存的满江红在分类学上有2个亚属,7个种1,对不同满江红叶腔中鱼腥藻的鉴别,多年来引起人们的重视.80年代,鱼腥藻的单克隆抗体和RFLP的研究,发现了它与宿主分类上一定程度的对应关系2,3.90年代以后,Van Coppenolle和Eskew分别以RAPD-PCR和DAF-PCR对从满江红共生体中提取的DNA进行了分析4,5,但后者发现了共生体中藻的DNA对整个PCR产物的干扰作用会影响满江红属本身系统分类.