Effects of ethyl pyruvate on leukocyte-endothelial interactions in the mesenteric microcirculation during early sepsis treatment

OBJECTIVES:Experimental studies on sepsis have demonstrated that ethyl pyruvate is endowed with antioxidant and anti-inflammatory properties. This study aimed to investigate the effects of ethyl pyruvate on leukocyte-endothelial interactions in the mesenteric microcirculation in a live Escherichia coli-induced sepsis model in rats.METHODS:Male Wistar rats were administered an intravenous suspension of E. coli bacteria or were subjected to a sham procedure. Three hours after bacterial infusion, the rats were randomized into the following groups: a control group without treatment, a group treated with lactated Ringer''s solution (4 mL/kg, i.v.), and a group treated with lactated Ringer''s solution (4 mL/kg, i.v.) plus ethyl pyruvate (50 mg/kg). At 24 h after bacterial infusion, leukocyte-endothelial interactions were investigated using intravital microscopy, and the expression of P-selectin and intercellular adhesion molecule-1 was evaluated via immunohistochemistry. White blood cell and platelet counts were also determined at baseline and 3 h and 24 h after E. coli inoculation.RESULTS:The non-treated and lactated Ringer''s solution-treated groups exhibited increases in the numbers of rolling leukocytes (∼2.5-fold increase), adherent cells (∼3.0-fold), and migrated cells (∼3.5-fold) compared with the sham group. In contrast, treatment with Ringer''s ethyl pyruvate solution reduced the numbers of rolling, adherent and migrated leukocytes to the levels observed in the sham group. Additionally, the expression of P-selectin and intercellular adhesion molecule-1 was significantly increased on mesenteric microvessels in the non-treated group compared with the sham group (p<0.001). The expression of both adhesion molecules was reduced in the other groups, with ethyl pyruvate being more effective than lactated Ringer''s solution. Infusion of bacteria caused significant leukopenia (3 h), followed by leukocytosis with granulocytosis (24 h). There was also an intense and progressive reduction in the number of platelets. However, no differences were observed after treatment with the different solutions.CONCLUSIONS:The presented data suggest that ethyl pyruvate efficiently reduces the inflammatory response in the mesenteric microcirculation in an experimental model of sepsis induced by live E. coli and is associated, at least in part, with down-regulation of P-selectin and intercellular adhesion molecule-1.     

E6/E7 oncoproteins of high risk HPV-16 upregulate MT1-MMP,MMP-2 and MMP-9 and promote the migration of cervical cancer cells

Background: E6 and E7 of high risk human papillomavirus 16 (HPV16) were reported to correlate with the cervical cancer (CC). And the presence of matrix metalloproteinases (MMPs) has also been indicated to be associated with CC. Methods: The present study investigated the expression of MMPs (MT1-MMP, MMP-2 and MMP-9) in CC cells with HPV16-E6/E7 oncoprotein(s) negative or positive, and then determined the regulation of HPV16-E6/E7 oncoproteins on the expression of MMPs (MT1-MMP, MMP-2 and MMP-9) and the migration of cervical cancer Caski and SiHa cells with RNAi technology. Results: It was demonstrated that the overexpression or the knockdown of HPV16-E6/E7 promoted or reduced MT1-MMP, MMP-2 and MMP-9 in CC cells. And the HPV16-E6, -E7 or -E6E7 influenced the migration of CC cells. The overexpression or the knockdown of them promoted or inhibited the migration of C33A or Caski/SiHa cells. Moreover, the chemical inhibition of MMP-2 or MMP-9 significantly reduced the migration of CC Caski or SiHa cells. Conclusions: Our results demonstrated that the E6-HPV16 or E7-HPV16 promoted the activity of MMP-2/9, and contributed to the migration of cervical cells.     

HPV-16 E6 promotes cell growth of esophageal cancer via downregulation of miR-125b and activation of Wnt/β-catenin signaling pathway

High-risk human papillomavirus (HPV) is a possible cause of esophageal cancer. However, the molecular pathogenesis of HPV-infected esophageal cancer remains unclear. The expression levels of some microRNAs including miR-125b have been negatively correlated with HPV infection, and miR-125b downregulation is associated with tumorigenesis. In addition, Wnt/β-catenin signaling pathway has been suggested to play an important role in esophageal cancer (EC). We examined miR-125b and Wnt/β-catenin signaling pathway in HPV-16 E6 promoted tumor progression in EC. HPV-16 E6 transfection decreased markedly the expression levels of miR-125b and promoted the colony formation in the Eca 109 and Kyse 150 cell lines, and restoration of miR-125b expression level antagonized the increased colony formation in HPV-16 E6 transfected cell lines. We also demonstrated that overexpression of E6 upregulated the Wnt/β-catenin signaling activity via modulating the multiple regulators including TLE1, GSK3β, and sFRP4. Overexpression of miR-125b restored the expression levels of these proteins. Expression of miR-125b was lower in HPV-16 E6 positive esophageal cancer tissues, and was negatively correlated with E6 mRNA levels. Our results indicate that HPV-16 E6 promotes tumorigenesis in EC via down-regulation of miR-125b, and this underlying mechanism may be involved in the activation of the Wnt/β-catenin signaling pathway.     

大黄对雌性幼年大鼠血清GnRH、LH、FSH、P、E2水平的影响

目的：：观察大黄水提物对雌性幼年大鼠血清促性腺激素释放激素(GnRH)、黄体生成素(LH)、卵泡刺激素(FSH)、孕酮(P)、雌二醇(E2)水平的影响。方法：4周龄雌性SD大鼠20只,随机分为正常对照组和大黄水提物2.0g/kg组,每组10只,连续灌胃给药30天。采用ELISA法检测各组大鼠血清GnRH、LH、FSH、P、E2的水平。结果：与正常对照组大鼠比较,大黄水提物2.0g/kg组大鼠GnRH、P、E2激素水平明显降低(P＜0.05),LH、FSH激素水平明显升高(P＜0.05)。结论：大黄水提物可明显影响雌性幼年大鼠下丘脑-垂体-卵巢轴的激素水平。     

不同戊型肝炎病毒IgG抗体检测试剂盒临床对比分析

目的比较磁微粒化学发光试剂与酶联免疫吸咐测定试剂检测戊型肝炎病毒Ig G抗体的结果差异。方法选取企业样本库2010年1月至2012年5月住院和门诊患者935例,分别用A厂家酶联免疫吸咐测定试剂和郑州安图生物磁微粒化学发光试剂测定戊型肝炎病毒Ig G抗体平行检测,不符样本用C厂家酶联免疫吸咐测定试剂复测。结果郑州安图生物磁微粒化学发光试剂和戊型肝炎病毒Ig G抗体A厂家酶联免疫吸咐测定试剂阳性一致百分比为95.5%,阴性一致百分比为97.7%,总符合率为97.2%,差异无统计学意义(P>0.05)。结论磁微粒化学发光戊型肝炎病毒Ig G抗体试剂与酶联免疫吸咐测定试剂临床性能相似,适于临床推广应用。     

The effect of vitamin D supplementation on cardiorespiratory fitness and muscle strength in male adults undergoing basic military training

BackgroundVitamin D deficiency (VDD) is ubiquitous in the Indian subcontinent. VDD has been shown to impair muscle functions. However, the association of VDD with cardiorespiratory endurance is uncertain. Hence, we enrolled and supplemented vitamin D in military recruits with VDD with an aim to evaluate effect of supplementation on cardiorespiratory endurance and muscle strength.MethodWe enrolled 90 military recruits with VDD and randomly allotted them to two groups equally. The group I received cholecalciferol granules 60,000 IU every fortnight for twelve weeks (cases), and the group II was observed as control. Muscle strength and cardiorespiratory endurance was assessed with a battery of tests (standing broad jump, bent arm hang test, 20 m shuttle run) at baseline and repeated at the end of training (nineteen weeks). Blood samples were collected for measurement of serum 25(OH)D and parathyroid hormone.ResultsIn Group I and Group II, there was significant increase in 25(OH) D levels (25.8 ± 7.1 and 17.3 ± 3.5 ng/ml, respectively), and in VO2 max (9.8 ± 8.8 and 12.7 ± 8.6 ml/kg/min, respectively) compared with the baseline values. However, no significant change was observed in muscle strength after supplementation. There was no difference between the groups in VO2 max and muscle strength at baseline and at the end of training.ConclusionVitamin D supplementation did not improve muscle strength and cardiorespiratory endurance in military recruits with VDD. A significant rise in 25(OH) D was observed even in those not supplemented with vitamin D.     

Impact of flexible insulin therapy on blood glucose variability,oxidative stress and inflammation in type 1 diabetic patients: The VARIAFIT study

AimsHbA_1c only partially predicts vascular risk in patients with type 1 diabetes (T1D), and a role for blood glucose variability (BGV) is a matter of debate. For this reason, this study investigated the impact of an educational programme of flexible insulin therapy (FIT) on BGV and oxidative stress.MethodsTests were conducted on 30 adult T1D patients in a prospective, single-centre trial at baseline (M0), and at 3 and 6 months (M3 and M6, respectively) of the FIT programme to determine BGV, as reflected by mean amplitude of glycaemic excursions (MAGE), low blood glucose index (LBGI), lability index (LI), average daily risk range (ADRR), glycaemic lability (scored by two diabetologists), urinary leukotriene E₄ (LTE4), 11-dehydro-thromboxane B₂ (TXB2) and 8-iso-prostaglandin F2α (PGF2).ResultsHbA_1c (7.7 ± 0.9%), ADRR, MAGE, LBGI and LI did not change from M0 to M3 and M6, although ADRR and LBGI significantly improved at M3 and M6 in patients with the highest baseline indices (≥ 40 and ≥ 5, respectively). TXB2 declined at M6 (832 ± 625 vs. 633 ± 972 pg/mg; P = 0.048), whereas LTE4 and PGF2 remained stable. ADRR showed the strongest correlation with glycaemic lability scores at all visits (r ≥ 0.84, P < 0.0001).ConclusionA FIT educational programme improved BGV only in patients with the highest baseline variability, and led to no changes in HbA_1c, while ADRR closely correlated with glycaemic lability score. Our data do not support a relationship between BGV and oxidative stress in T1D patients, although the impact of variability on TXB2 deserves further investigation (ClinicalTrials.gov NCT00973492).     

白藜芦醇对大肠杆菌诱导的大鼠膀胱粘膜损伤保护作用机制的研究

目的 探讨白藜芦醇对大肠杆菌诱导的大鼠膀胱粘膜损伤的保护作用机制。 方法 将 45 只(170±10) g 雌性 SD 大鼠随机分为空白对照组、模型对照组和白藜芦醇实验组。 通过尿道向大鼠膀胱内植入导管并进行大肠杆 菌液灌注建立感染性膀胱粘膜损伤模型(模型对照组),白藜芦醇实验组按照 40 mg / kg 剂量灌胃给药,实验结束后 镜检观察大鼠膀胱内壁病理变化。 采用比色法检测超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧 化氢酶(CAT)、一氧化氮(NO)和丙二醛(MDA)含量;采用 ELISA 法检测膀胱组织内细胞因子白介素-1β( IL-1β)、 IL-6、IL-8、IL-10 和肿瘤坏死因子(TNF-α)的含量;采用 Western Blot 法检测转化生长因子-β1(TGF-β1)的表达。 结果 白藜芦醇给药显著减少膀胱黏膜中炎性细胞数量,显著降低大鼠膀胱组织中 MDA、NO 含量,提高了 SOD、GSH -Px、CAT 活性;明显降低 IL -1β、IL-8、TNF-α,提高 IL-6、IL-10 含量;显著减少 TGF-β1 的蛋白表达。 结论 白藜 芦醇通过抗氧化、抗炎和降低纤维化作用改善大肠杆菌诱导的大鼠膀胱粘膜损伤,避免对膀胱组织伤害。     

An ORF1‐rearranged hepatitis E virus derived from a chronically infected patient efficiently replicates in cell culture

Hepatitis E is an increasingly reported disease in industrialized countries. Studies on the replication cycle of hepatitis E virus (HEV) are hampered due to the lack of efficient and robust cell culture systems for this virus. We describe the successful isolation of HEV derived from a chronically infected kidney transplant patient held under immunosuppressive therapy. Inoculation of serum sample 47832 onto the human lung carcinoma cell line A549 resulted in the replication of the virus as shown by RT‐qPCR. This novel human‐derived HEV strain is closely related to a wild boar‐derived genotype 3 strain, which did not replicate in A549 cells. It carries a 186 nucleotide insertion in the hypervariable ORF1‐region, derived from two parts of its ORF1. By passaging of the infected cells, a cell line continuously producing HEV particles was generated as demonstrated by RT‐qPCR, immuno‐electron microscopy, density gradient centrifugation and immunohistochemistry. Replication of the produced virus was demonstrated after its inoculation onto fresh A549 cells and two consecutive passages, whereas heating at 65 °C for 2 min abolished its infectivity. Several point mutations scattered along the whole genome were present in the HEV strain from the second passage; however, the ORF1 insertion was still present. Previously, cell culture isolation of two other HEV strains carrying insertions in their hypervariable regions, but originating from human ribosomal protein genes, has been described. The findings may indicate that cell culture adaptation of is mostly dependent on the length and position of the insertion, rather than from the sequence itself.