Angiopoietin decoy secreted at tumor site impairs tumor growth and metastases by inducing local inflammation and altering neoangiogenesis

The extracellular domain of the receptor tyrosine kinase Tie2/TEK (exTEK) has been used as an angiopoietin decoy to study the role of angiopoietins in the tumor–host interactions, using a syngeneic model of experimental metastases and subcutaneous tumor. Soluble exTEK secreted by transfected tumor cells inhibited HUVECs from forming tubes in Matrigel. ExTEK-transfected C26 colon carcinoma and TS/A mammary tumor cells displayed reduced growth rate when injected subcutaneously, and reduced ability to form experimental metastases when injected intravenously. Immunohistochemical analysis of tumors and metastases showed increased leukocytes infiltration and signs of inflammation in exTEK-secreting compared to parental tumor, as well as impairment in neo-vessel growth and organization. However, while neoangiogenesis eventually rescued in the subcutis, it failed to organize in the experimental metastases of exTEK-secreting tumor, contributing to the hampering of metastatic growth and to increased mice survival. The reactive infiltrate of C26TEK contained a different percentage of leukocytes and was responsible for the tumor inhibition. In fact, leukopenia induced by -irradiation of recipient mice or injection into interferon gamma (IFN-) gene knockout (GKO) mice resulted in reduced mouse survival and an increased number of lung metastases. On the other hand, interleukin (IL)-12 treatment prolonged the survival of mice bearing subcutaneous C26TEK but not of those bearing lung metastases, suggesting that IL-12 could exert further antiangiogenic effects at the site where the tumor can restore neoangiogenesis. These results show in vivo that reduced angiopoietin availability at the tumor site induces a local inflammatory response and impairment of neoangiogenesis which act synergistically to limit tumor growth and metastasis.Abbreviations AEC amino-ethylcarbazole - ELISA enzyme-linked immunosorbent assay - HRP horseradish peroxidase - HUVEC human umbilical vascular endothelial cell - i.v. intravenous - s.c. subcutaneous - TBS Tris-HCl buffered solution     

Infiltration with <Emphasis Type="Italic">Agrobacterium</Emphasis> — the method for stable transformation avoiding tissue culture

A number of in planta transformation protocols that avoid long culture under sterile conditions were developed for Arabidopsis thaliana. The most widely used methods are based on vacuum infiltration and floral dip. These methods were adapted for transformation of other species as well. Successful in planta transformations of alfalfa, radish, pakchoi and petunia were reported recently. In this short review we present several modified procedures originally developed for Arabidopsis thaliana and in some cases adapted to other species. We emphasize the crucial parameters involved in in planta transformation. We also describe here the studies attempting to shed light on the mechanisms and estimating the cellular target of transformation, which may help in transforming new plant species.     

Enhancing effect of dietary cholesterol and inhibitory effect of pravastatin on allergic pulmonary inflammation

To elucidate the link between the intake of animal fat and asthma, a murine model was developed to examine the effect of dietary cholesterol on pulmonary allergic inflammation. Male C57BL6 mice were fed either a control diet or a diet supplemented with 2% cholesterol. Following sensitization and inhalation exposure to ovalbumin, the bronchoalveolar lavage fluid of mice in the cholesterol group contained higher numbers of eosinophils and elevated levels of IL-5, PGE₂, and MCP-1. In addition, dietary cholesterol also resulted in elevated production of IL-4 and IFN- by lymphocytes isolated from the lungs. These inflammatory indicators were all significantly correlated with serum cholesterol levels. In contrast to the effect of dietary cholesterol, adding pravastatin to the drinking water significantly reduced eosinophil infiltration and the levels of IL-5, PGE₂ and MCP-1 in lavage fluid. Although dietary cholesterol did not alter baseline IL-12 in the lungs, in mice challenged with ovalbumin the IL-12 levels were reduced in the cholesterol group and elevated significantly in the pravastatin group. The results suggest that dietary cholesterol might enhance pulmonary allergic inflammation, possibly involving both nonspecific inflammatory processes and lymphocyte activities.     

Teprenone, but not H2-receptor blocker or sucralfate, suppresses corpus Helicobacter pylori colonization and gastritis in humans: teprenone inhibition of H. pylori-induced interleukin-8 in MKN28 gastric epithelial cell lines

Background. The role of teprenone in Helicobacter pylori‐associated gastritis has yet to be determined. To investigate the effect of teprenone on inflammatory cell infiltration, and on H. pylori colonization of the gastric mucosa in H. pylori‐infected patients, we first compared the effect of teprenone with that of both histamine H2 receptor antagonists (H2‐RA) and sucralfate on the histological scores of H. pylori gastritis. We then examined its in vitro effect on H. pylori‐induced interleukin (IL)‐8 production in MKN28 gastric epithelial cells. Materials and Methods. A total of 68 patients were divided into three groups, each group undergoing a 3‐month treatment with either teprenone (150 mg/day), H2‐RA (nizatidine, 300 mg/day), or sucralfate (3 g/day). All subjects underwent endoscopic examination of the stomach before and after treatment. IL‐8 production in MKN28 gastric epithelial cells was measured by enzyme‐linked immunosorbent assay (ELISA). Results. Following treatment, the teprenone group showed a significant decrease in both neutrophil infiltration and H. pylori density of the corpus (before vs. after: 2.49 ± 0.22 vs. 2.15 ± 0.23, p = .009; 2.36 ± 0.25 vs. 2.00 ± 0.24, p = .035, respectively), with no significant differences seen in either the sucralfate or H2‐RA groups. Teprenone inhibited H. pylori‐enhanced IL‐8 production in MKN28 gastric epithelial cells in vitro, in a dose‐dependent manner. Conclusions. Teprenone may modify corpus H. pylori‐associated gastritis through its effect on neutrophil infiltration and H. pylori density, in part by its inhibition of IL‐8 production in the gastric mucosa.     

森林流域土壤饱和渗透系数与有效孔隙度模型的研究

以长白山自然保护区内未受扰动的两种典型原始森林土壤为对象,通过野外选取原状土样,在实验室内测定其饱和渗透系数和有效孔隙度,利用回归分析法分别建立了森林土壤内饱和渗透系数和有效孔隙度随深度变化的数学模型,即Ｋｓ（ｚ）＝Ｋ０－ｆ１ｌｎａ１ｚ和ω（ｚ）＝ω０－ｆ２ｌｎａ２ｚ．将所得出的对数模型与Ｂｅｖｅｎ所提出的指数模型进行比较表明,在森林流域内对数模型不仅适用范围比指数模型广,而且精度明显高于指数模型,它更接近于野外实际情况．     

The effects of either resveratrol or exercise on macrophage infiltration and switching from M1 to M2 in high fat diet mice

[Purpose]

The aim of this study was to compare the effectiveness of either resveratrol supplementation or exercise training on macrophage infiltration and switching from M1 to M2 kupffer cells in high fat diet mice.

[Methods]

C57BL/6 mice were separated into 5 groups: normal diet (ND; n = 6), high-fat diet (HD; n = 6), high-fat diet with resveratrol (HR; n = 6), high-fat diet with exercise (HE; n = 6) or high-fat diet with resveratrol and exercise (HRE; n = 6). Resveratrol supplementation mice were orally gavaged with resveratrol (25mg/kg of body weight) dissolved in 50% propylene glycol. Exercise mice ran on a treadmill at 12-20 m/min for 30-60 min/day, 5 times/week for 12 weeks.

[Results]

After 12 weeks of intervention, the liver was analyzed. F4/80 expression was evaluated by western blot while CD11c and CD163 mRNA expressions were evaluated by RT-PCR. The weights of the body and liver were significantly increased in the HD and HR group compared to the ND group (p < 0.01). However, the weights were most effectively reduced in the HE and HRE groups compared to the HD group (p < 0.05). The macrophage marker, F4/80 expression was significantly lower in the HE and HRE groups compared to the HD group (p < 0.05). mRNA expression of the M1 macrophage marker, CD11c, in the HD group was significantly increased compared to the ND group (p < 0.01). mRNA expression of the M2 macrophage specific marker, CD163, in the HE and HRE groups were significantly increased compared to the HD group (p < 0.05). The mRNA expressions of TLR4, ICAM-1 and VCAM-1, which induce pro-inflammatory cytokine production, were strongly decreased in the HR, HE, and HRE groups compared to the HD group.

[Conclusion]

These results suggest that moderate exercise training inhibits macrophage infiltration and up regulation of CD163 expression. However, resveratrol supplementation is not enough to ameliorate obesity-induced macrophage infiltration and switching.     

Column-switching HPLC–MS/MS analysis of ropivacaine in serum,ultrafiltrate and drainage blood for validating the safety of blood reinfusion

A high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS–MS) method, using back-flush column-switching was developed for total drug concentrations of ropivacaine in serum and drainage blood in the measuring range 0.1–10 μg/mL. Samples were diluted with internal standard (²H₇-ropivacaine) and extraction buffer, centrifuged and injected directly onto a BioTrap 500 MS extraction column. Using a time programmed six-port valve switch, ropivacaine was back-flushed onto a Zorbax SB-Aq analytical column, gradient eluted and finally detected after electro spray ionisation and multiple reaction monitoring (MRM) of the transitions m/z 275 → m/z 126 and m/z 282 → m/z 133 for ropivacaine and ²H₇-ropivacaine, respectively. Accuracy (bias-%) was −1.5 to 5.8% and intermediate precision (C.V.) was 1.4–3.1%. The low sample amount required (10 μL), high specificity and short run time (6 min) makes it very suitable for determination of ropivacaine. Using the same methodology as described above and 200 μL ultrafiltrate, the free drug concentrations of ropivacaine in serum could be precisely determined with a C.V. below 3%. The method was used to investigate the safety of reinfusion of drainage blood after knee and hip arthroplasty when ropivacaine (Naropin^®) was used for local analgesia. Data for 30 patients are summarised.     

能谱CT成像对甲状腺癌局部浸润深度的诊断价值及其定量参数与肿瘤组织中Ki67、VEGF、CD34、EGFR的相关性

摘要 目的：探讨能谱CT成像对甲状腺癌局部浸润深度的诊断价值及其定量参数与肿瘤组织中Ki67、VEGF、CD34、EGFR的相关性。方法：回顾性分析2018年6月-2021年6月我院经手术或穿刺活检病理证实为甲状腺肿瘤性病变的患者96例，其中29例为甲状腺癌局部浸润组（A组），34例为甲状腺癌无浸润组（B组），33例为甲状腺腺瘤组（C组）。另取56例甲状腺另一侧叶正常组织作为对照组（D组）。所有患者均完善能谱CT检查，采集图像后在能谱CT Viewer分析软件上测量病变区碘浓度，计算能谱曲线斜率。采用免疫组织化学染色分析Ki-67、VEGF、CD34、EGFR的表达情况。采用Spearman秩相关分析评价碘浓度、能谱曲线斜率与甲状腺癌肿瘤组织中Ki-67、VEGF、CD34、EGFR表达的相关性。结果：在平扫、动脉期、静脉期，A组、B组、C组和D组的碘浓度逐渐增大，两两比较差异均有统计学意义（P＜0.05）。甲状腺癌局部浸润组织能谱曲线呈"低平型"，斜率为较小负值，正常甲状腺组织能谱曲线为下降型，斜率为负值；在平扫、动脉期、静脉期，A组、B组、C组和D组的能谱曲线斜率逐渐变小，差异均有统计学意义（P＜0.05）。A组中Ki-67、VEGF、CD34和EGFR的阳性表达率均高于B组和C组，差异均有统计学意义（P＜0.05）。碘浓度在动脉期、静脉期与Ki-67、VEGF、CD34、EGFR表达呈正相关（P＜0.05），碘浓度在平扫与Ki-67表达呈正相关（P＜0.05），碘浓度在平扫与VEGF、CD34、EGFR表达无相关性（P＞0.05）。能谱曲线斜率在动脉期、静脉期与Ki-67、VEGF、CD34、EGFR表达呈正相关（P＜0.05），能谱曲线斜率在平扫与VEGF表达呈正相关（P＜0.05），能谱曲线斜率在平扫与Ki-67、CD34、EGFR表达无相关性（P＞0.05）。结论：能谱CT成像检查对甲状腺癌局部浸润深度的判断具有重要的价值，其定量参数碘浓度、能谱曲线斜率与Ki67、VEGF、CD34、EGFR具有相关性，可间接反映肿瘤微血管、肿瘤血管生成、甲状腺癌分化程度、浸润程度等情况，对评价甲状腺癌生物学行为可提供有价值的信息。     

自然降雨下黄土丘陵区草灌植物垂直覆盖结构的减流减沙效应

黄土高原是我国水土流失和生态环境问题最为严重的地区之一,植被恢复是防治水土流失的重要措施。植物垂直覆盖结构包括地上冠层、地表枯落物和地下根系,各组分具有不同的水土保持作用,是研究植被与水土流失关系的基本单元。目前,关于植物垂直覆盖结构不同组分对土壤侵蚀影响的研究主要是基于人工模拟降雨,缺少自然降雨条件下不同植物的垂直覆盖结构对产流、产沙和入渗等多过程影响的系统研究。本研究以黄土丘陵区典型的草本(须芒草)、半灌木(铁杆蒿)和灌木(绣线菊)为研究对象,每种植物进行三种处理(自然状态、去除枯落物和仅留根系)以及裸地对照,观测2015—2016年降雨事件的产流产沙量和入渗量,分析植物不同垂直覆盖结构的减流减沙效益及其相对贡献。结果表明:三种植物均具有较好的减流(45.9%—73.2%)、减沙效益(87.5%—94.6%)和增加入渗作用(4.7%—10.8%),灌木的减流效果(73.2%)显著高于草本(45.9%)和半灌木(63.5%),但三种植物间的减沙效益没有显著性差异。冠层的减流作用最大,贡献率接近一半(48%—50%),草本枯落物的减流贡献率与根系基本一致,而半灌木和灌木枯落物的减流贡献...     

超声诊断胰腺脂肪浸润与血脂、脂肪肝的关系研究

目的：探讨超声诊断胰腺脂肪浸润与血脂、脂肪肝的关系。方法：回顾性分析63例胰腺脂肪浸润者与148例同期检查无胰腺脂肪浸润者的病历资料,研究胰腺脂肪浸润的超声特点以及与脂肪肝、血脂浓度、年龄、血压等的关系。结果：胰腺脂肪浸润超声显示为胰腺饱满,边缘模糊,回声增强且明显强于肝脏,与正常胰腺超声影像容易鉴别。胰腺脂肪浸润组血浆甘油三酯浓度2．17±1．22mmol／L,比非浸润组的1．35±1．06mmol／L明显高（P〈0．05）,胰腺脂肪浸润组患者100％（63／63）合并脂肪肝明显高于非浸润组的20．95％（31／148）（P〈0．05）。二组在胆固醇浓度、高血压发病率及年龄方面无显著差异（P〉0．05）。结论：胰腺脂肪浸润与脂肪肝、血浆甘油三酯浓度有着一定的相关性,超声诊断胰腺脂肪浸润对于提示其他相关疾病与异常的存在具有重要临床参考价值。