In Vitro and In Vivo Activities of Tilmicosin and Acetylisovaleryltylosin Tartrate against Toxoplasma gondii

Toxoplasma gondii is a widespread intracellular pathogen that infects humans and a variety of animals. The current therapeutic strategy for human toxoplasmosis is a combination of sulphadiazine and pyrimethamine. However, this combination still has a high failure rate and is ineffective against chronic infections. Therefore, it is important to discover a new anti-T. gondii drug that is safer and more effective in both humans and animals. In this study, we describe the anti-T. gondii activities of the 16-membered macrolide tilmicosin and acetylisovaleryltylosin tartrate (ATLL). Both tilmicosin and ATLL potently inhibited T. gondii with a half-maximal effective concentration (EC₅₀) of 17.96 μM and 10.67 μM, respectively. Interestingly, tilmicosin and ATLL had different effects on the parasites. ATLL exhibited a potent inhibitory effect on intracellular parasite growth, while tilmicosin suppressed parasites extracellularly. By studying the lytic cycle of T. gondii after treatment, we found that ATLL potently inhibited the intracellular proliferation of tachyzoites, while tilmicosin affected the invasion of tachyzoites. Immunofluorescence analysis using ATLL-treated T. gondii showed morphologically abnormal parasites, which may be due to the inhibition of tachyzoite proliferation and division. In addition, tilmicosin and ATLL significantly delayed the death of mice caused by acute toxoplasmosis. Our results suggest that ATLL has potent anti-Toxoplasma activity both in vitro and in vivo and may be an alternative to toxoplasmosis in the future.     

重组弓形虫热休克蛋白70联合弗氏佐剂免疫小鼠的抗弓形虫感染作用

目的评价重组弓形虫热休克蛋白70(rTgHSP70)联合弗氏佐剂皮下免疫小鼠的抗弓形虫感染作用。方法BALB/c小鼠70只,随机分为7组,分别为PBS组、A组(佐剂+PBS)、20PA组(20μg rTgHSP70+佐剂)、40P组(40μgrTgH-SP70)、40PA组(40μg rTgHSP70+佐剂)、60PA组(60μg rTgHSP70+佐剂)及80PA组(80μg rTgHSP70+佐剂),皮下免疫,间隔2周,加强免疫2次,共免疫3次。末次免疫后14d,用2×104个RH株弓形虫速殖子/只灌胃攻击各组小鼠,攻击后30d处死,ELISA检测血清IgG和肠液sIgA;分离脑组织内弓形虫速殖子,并计数。结果在添加佐剂组中,血清IgG水平随rTgHSP70剂量的增加而升高,与PBS组和A组比较,差异有统计学意义;肠液sIgA水平60PA组最高,与20PA和40PA组比较,差异有统计学意义;与PBS组相比,A、20PA、40P、40PA、60PA和80PA组的脑组织内速殖子数分别减少了56.87%、56.46%、61.90%、48.74%、62.87%和70.44%。结论rTgHSP70联合弗氏佐剂皮下免疫小鼠可诱导一定的抗弓形虫感染作用,80μgrTgHSP70为较佳剂量。     

重组弓形虫热休克蛋白70不同途径免疫小鼠诱导的小肠黏膜IgA抗体分泌细胞及sIgA抗体的动态变化

目的分析重组弓形虫热休克蛋白70(rTgHSP70)鼻内或皮下免疫小鼠诱导的小肠黏膜IgA抗体分泌细胞(IgA antibody-secreting cells,IgA-ASCs)与小肠冲洗液sIgA抗体的动态变化及二者的相关性,探讨其上调小肠黏膜免疫应答的作用机制。方法 BALB/c小鼠90只,随机均分3组:鼻内组(20μg rTgHSP70/只,滴鼻,免疫2次,间隔2周)、皮下组(80μg rTgHSP70/只,背部皮下注射,免疫2次,间隔2周)、对照组(不免疫)。分别于末次免疫后第1、2、3、4、5、6周,每组取小鼠5只,脱颈椎处死,免疫组化法检测十二指肠、空肠及回肠黏膜IgA-ASCs数量,ELISA法检测小肠冲洗液sIgA水平。结果 IgA-ASCs分布于小肠黏膜的固有层中,且鼻内组小肠黏膜IgA-ASCs数量明显高于皮下组和对照组(P<0.001),鼻内组小肠冲洗液sIgA水平在免后1～4周均处于上升阶段,第4周达到高峰,显著高于皮下组和对照组,差异有统计学意义(P<0.001);鼻内组和皮下组小肠冲洗液sIgA水平与十二指肠、空肠、回肠黏膜固有层IgA-ASCs的数量呈正相关(P<0.05)。结论 rTgHSP70经鼻内或皮下免疫小鼠均可诱导小肠黏膜固有层IgA-ASCs和小肠冲洗液sIgA水平的持续高表达,且二者呈正相关,鼻内免疫上调小肠黏膜免疫的作用优于皮下免疫。     

新城疫病毒自然宿主细胞膜受体的鉴定

目的鉴定鸡源副黏病毒F48E9株的自然宿主细胞膜受体。方法采用蛋白膜提取试剂盒提取鸡胚成纤维细胞膜蛋白,用改进的间接ELISA方法检测鸡源副黏病毒分离株F48E9与细胞膜的结合活性,利用病毒铺覆蛋白印迹技术(VOPBA)鉴定新城疫病毒(NDV)的自然宿主细胞膜受体。结果NDV与鸡胚成纤维细胞膜有很强的结合力,在转印鸡胚成纤维细胞膜蛋白的PVDF膜上有几条明显的疑似受体条带,相对分子质量介于35000～60000之间。结论初步鉴定了NDV自然宿主细胞膜的受体,其疑似受体蛋白的性质及在新城疫病毒致病中的作用尚有待进一步研究。     

The Transport of Copper(II) through Hollow Fiber Renewal Liquid Membrane and Hollow Fiber Supported Liquid Membrane

Abstract

In this paper, hollow fiber renewal liquid membrane (HFRLM) and hollow fiber supported liquid membrane (HFSLM) were used to simultaneously remove and recover copper(II) from aqueous solutions, and the transport performance of these two techniques were compared under the similar conditions for the system of CuSO₄ +D2EHPA in kerosene +HCl. The results showed that the HFRLM process was more stable than the HFSLM process. The HFRLM process had a higher overall mass transfer coefficient than that of HFSLM process in single-pass experiments. These were because the renewal effect of the liquid membrane layer could reduce the mass transfer resistance of the lumen side and replenish the loss of the membrane liquid in the HFRLM process. The transport results were better in the HFRLM process than that in the HFSLM process with recycling experiments. Therefore, HFRLM technique is a promising method for simultaneous removal and recovery of heavy metal from aqueous solutions.     

Prostaglandin E2 Receptor 4 (EP4) Affects Trophoblast Functions via Activating the cAMP-PKA-pCREB Signaling Pathway at the Maternal-Fetal Interface in Unexplained Recurrent Miscarriage

Implantation consists of a complex process based on coordinated crosstalk between the endometrium and trophoblast. Furthermore, it is known that the microenvironment of this fetal–maternal interface plays an important role in the development of extravillous trophoblast cells. This is mainly due to the fact that tissues mediate embryonic signaling biologicals, among other molecules, prostaglandins. Prostaglandins influence tissue through several cell processes including differentiation, proliferation, and promotion of maternal immune tolerance. The aim of this study is to investigate the potential pathological mechanism of the prostaglandin E2 receptor 4 (EP4) in modulating extravillous trophoblast cells (EVTs) in unexplained recurrent marriage (uRM). Our results indicated that the expression of EP4 in EVTs was decreased in women experiencing uRM. Furthermore, silencing of EP4 showed an inhibition of the proliferation and induced apoptosis in vitro. In addition, our results demonstrated reductions in β- human chorionic gonadotropin (hCG), progesterone, and interleukin (IL)-6, which is likely a result from the activation of the cyclic adenosine monophosphate (cAMP)- cAMP-dependent protein kinase A (PKA)-phosphorylating CREB (pCREB) pathway. Our data might provide insight into the mechanisms of EP4 linked to trophoblast function. These findings help build a more comprehensive understanding of the effects of EP4 on the trophoblast at the fetal–maternal interface in the first trimester of pregnancy.     

聚醚共聚酰胺(PEBA)膜的溶胀和渗透汽化行为研究

用涂布法制备出PEBA均质膜,分别测试了膜在纯醋酸正丁酯、水以及醋酸正丁酯稀水溶液中的溶胀率,并考察了浸泡时间、温度和溶液浓度对溶胀率的影响;结果表明:PEBA膜在水中溶胀很小,对醋酸正丁酯具有优先吸附能力;溶胀率随浸泡时间的延长、浸泡液浓度和温度的升高而逐渐上升。渗透汽化实验表明膜对醋酸正丁酯的良好选择性,膜的渗透通量和分离因子随着浓度的增加而增加,渗透通量随着温度的升高而增加,但分离因子随温度的升高而减小;当原料液浓度为0.6%(wt)时,醋酸正丁酯的渗透通量为143.9g·m-2·h-1,分离因子达到236.9。     

Aryl Hydrocarbon Receptor Defect Attenuates Mitogen-Activated Signaling through Leucine-Rich Repeats and Immunoglobulin-like Domains 1 (LRIG1)-Dependent EGFR Degradation

Aryl hydrocarbon receptor (AHR) genomic pathway has been well-characterized in a number of respiratory diseases. In addition, the cytoplasmic AHR protein may act as an adaptor of E3 ubiquitin ligase. In this study, the physiological functions of AHR that regulate cell proliferation were explored using the CRISPR/Cas9 system. The doubling-time of the AHR-KO clones of A549 and BEAS-2B was observed to be prolonged. The attenuation of proliferation potential was strongly associated with either the induction of p27^Kip1 or the impairment in mitogenic signal transduction driven by the epidermal growth factor (EGF) and EGF receptor (EGFR). We found that the leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1), a repressor of EGFR, was induced in the absence of AHR in vitro and in vivo. The LRIG1 tends to degrade via a proteasome dependent manner by interacting with AHR in wild-type cells. Either LRIG1 or a disintegrin and metalloprotease 17 (ADAM17) were accumulated in AHR-defective cells, consequently accelerating the degradation of EGFR, and attenuating the response to mitogenic stimulation. We also affirmed low AHR but high LRIG1 levels in lung tissues of chronic obstructive pulmonary disease (COPD) patients. This might partially elucidate the sluggish tissue repairment and developing inflammation in COPD patients.     

蚕蛹蛋白分离膜的探索研究

本研究以聚乙烯醇（PVA）为膜材料，以聚乙二醇（PEG）为致孔剂配制铸膜液，凝固浴为去离子水，利用相转化法制备超滤（UF）膜用于分离蚕蛹蛋白。实验中初步研究了不同分子量的PEG对成膜的影响：不同比例的PVA／PEG-1000条件下．不同PVA成型浓度所成膜对蚕蛹蛋白的截留情况。结果显示PEG-1000对PVA—PEG超滤膜的改性有较好的作用。得到无色透明、机械强度较高的膜。当PVA／PEG-1000≤9时。得到的是半透明膜，而在PVA／PEG-1000≥9时，得到的膜透明度高；当PVA浓度不同时成型，即使膜的成分相同，膜的性能也有很大的不同。在PVA的浓度为5％时，膜的截留率在PVA／PEG-1000=12时最好。达83．47％。在PVA的浓度为10％时，膜的截留率在PVA／PEG-1000=3时最好，达83．76％；在PVA的浓度为15％时，其截留率比较高，虽都没有超过80％。但比较稳定。     

Controllable Fabrication of CuO/Ammonium Perchlorate (AP) Nanocomposites through Ceramic Membrane Anti‐Solvent Recrystallization

To improve the dispersion of CuO nanoparticles in ammonium perchlorate (AP), CuO/AP nanocomposites are designed and the novel ceramic membrane anti‐solvent crystallization (CMASC) method is applied to the nanocomposites synthesis. Typical experimental results demonstrate that nanocomposites with a size of less than 20 μm exhibit well‐defined hexahedral‐like structure and that the CuO nanoparticles are physically coated by AP crystals. The well‐dispersion of CuO nanoparticles and preparation of superfine AP crystals can be achieved at one step. The nucleation and growth mechanism of nanocomposites is discussed. To explore the actual formation process of the nanocomposites, relevant experiments are designed and carried out. The results are well consistent with previous assumptions and verify that several parameters, including feeding rate, volume ratio of matched solvent and anti‐solvent, can be employed to manipulate the size and morphology of the nanocomposites. The catalytic activity of CuO nanoparticles in the nanocomposites exhibits superior performance.     

配合物{[Co（HFlu）2Cl2】·2C2H5OH}n的合成、结构及体外抗真菌活性

新合成了1种钴-氟康唑配合物{[Co（HFlu）2Cl2】·2C2H5OH}n，利用X-射线单晶衍射仪等对配合物进行了结构分析，并研究了该配合物的体外抗真菌活性，标题化合物对丝状真菌的抗菌效果明显优于氟康唑，对酵母菌的体外抗真菌活性与氟康唑相当，     

Propylene/1‐Hexene Copolymer as a Tailor‐Made Poly(propylene) for Membrane Preparation via the Thermally Induced Phase Separation (TIPS) Process

Summary: Poly[propylene‐co‐(1‐hexene)], one example of a “tailor‐made poly(propylene)”, was synthesized using an iso‐specific metallocene catalyst in order to study the influence of copolymer composition on the pore size of isotactic poly(propylene) (iPP) membranes prepared by the TIPS process. The structure of the copolymers and their properties in solution were analyzed and discussed in relation to the polymer‐diluent phase diagram, the droplet growth kinetics during the TIPS process, the viscosity of the system and the final pore size of the membranes. The crystallization curve in the phase diagram was found to shift significantly as comonomer content increased and thus the droplet growth period was drastically increased. The resulting increase of the characteristic pore size in the membranes demonstrated that it is possible to use tailor‐made poly(propylene)s to control the pore size in porous membranes prepared via the TIPS process (under otherwise constant conditions).

Porous size is controlled by the polymer and the TIPS process.     

Role of Receptor Protein Tyrosine Phosphatases (RPTPs) in Insulin Signaling and Secretion

Changes in lifestyle in developed countries have triggered the prevalence of obesity and type 2 diabetes mellitus (T2DM) in the latest years. Consequently, these metabolic diseases associated to insulin resistance, and the morbidity associated with them, accounts for enormous costs for the health systems. The best way to face this problem is to identify potential therapeutic targets and/or early biomarkers to help in the treatment and in the early detection. In the insulin receptor signaling cascade, the activities of protein tyrosine kinases and phosphatases are coordinated, thus, protein tyrosine kinases amplify the insulin signaling response, whereas phosphatases are required for the regulation of the rate and duration of that response. The focus of this review is to summarize the impact of transmembrane receptor protein tyrosine phosphatase (RPTPs) in the insulin signaling cascade and secretion, and their implication in metabolic diseases such as obesity and T2DM.     

Squamosamide Derivative FLZ Protects Retinal Pigment Epithelium Cells from Oxidative Stress through Activation of Epidermal Growth Factor Receptor (EGFR)-AKT Signaling

Reactive oxygen species (ROS)-mediated retinal pigment epithelium (RPE) cell apoptosis is attributed to age-related macular degeneration (AMD) pathogenesis. FLZ, a novel synthetic squamosamide derivative from a Chinese herb, Annona glabra, has displayed significant cyto-protective activity. In the current study, we explored the pro-survival effect of FLZ in oxidative stressed-RPE cells and studied the underlying signaling mechanisms. Our results showed that FLZ attenuated hydrogen peroxide (H₂O₂)-induced viability decrease and apoptosis in the RPE cell line (ARPE-19 cells) and in primary mouse RPE cells. Western blotting results showed that FLZ activated AKT signaling in RPE cells. The AKT-specific inhibitor, MK-2206, the phosphoinositide 3-kinase (PI3K)/AKT pan inhibitor, wortmannin, and AKT1-shRNA (short hairpin RNA) depletion almost abolished FLZ-mediated pro-survival/anti-apoptosis activity. We discovered that epidermal growth factor receptor (EGFR) trans-activation mediated FLZ-induced AKT activation and the pro-survival effect in RPE cells, and the anti-apoptosis effect of FLZ against H₂O₂ was inhibited by the EGFR inhibitor, PD153035, or by EGFR shRNA-knockdown. In conclusion, FLZ protects RPE cells from oxidative stress through activation of EGFR-AKT signaling, and our results suggest that FLZ might have therapeutic values for AMD.     

Separation of Silver(I) and Copper(II) from Aqueous Solutions by Transport through Polymer Inclusion Membranes with Cyanex 471X

In this work the selective transport of silver(I) and copper(II) ions from aqueous nitrate(V) solutions by transport through polymer inclusion membrane (PIM) has been studied. The membrane consisted of cellulose triacatate (CTA) as the polymeric support, o-nitrophenyl pentyl ether (ONPPE) as the plasticizer and Cyanex 471X (triisobutylphosphine sulphide) as the ion carrier. Ag(I) ions were effectively removed from the source phase by transport through PIM into 0.01 M Na₂S₂O₃ as the receiving phase. The influence of membrane composition on the transport of silver(I) ions has been evaluated.     

Selective Transport of Cu(II) across a Polymer Inclusion Membrane with 1-Alkylimidazole from Nitrate Solutions

The selective transport of copper(II), zinc(II), cobalt(II), and nickel(II) ions from nitrate solutions across polymer inclusion membranes (PIMs), which consist of cellulose triacetate as polymeric support, o-nitrophenyl pentyl ether as plasticizer, and 1-alkylimidazole (alkyl from hexyl- to decyl) as ion carrier was reported. PIM was characterized by using atomic force microscopy (AFM) technique. The results show that Cu(II) can be separated very effectively from other transition metal cations as Zn(II), Co(II), and Ni(II) (at a concentration of 10^?3 mol/dm³ each). Alkyl substituents at position 1 of the imidazole ring have been found to affect the hydrophobic properties and initial flux of the transported metal ions. The efficiency of separation of metal ions by 1-alkylimidazole followed the sequence: Cu(II) > Zn(II) > Co(II) > Ni(II). The highest selectivity coefficient for Cu(II) was found with 1-hexylimidazole and its 1 mol/dm³ solution in PIM. Separation of the ions was more effective for the nitrates(V) than for chlorides.     

Preparation and Characterization of Fe2O3/Ammonium Perchlorate (AP) Nanocomposites through Ceramic Membrane Anti‐Solvent Crystallization

A novel ceramic membrane anti‐solvent crystallization (CMASC) method was proposed to prepare Fe₂O₃/AP nanocomposites with core‐shell structure. For the preparation of Fe₂O₃/AP nanocomposites, several key advantages of the CMASC method are as follows. Firstly, both well‐dispersed Fe₂O₃ nanoparticles and the superfine AP preparation can be achieved at one step. Secondly, no non‐component of solid propellant was involved in this composite process. Thirdly, the size and morphology of Fe₂O₃/AP nanocomposites can be effectively controlled by using the ceramic membrane with regular pore structure as feeding template. The morphology and structure of Fe₂O₃/AP nanocomposites were characterized by inductively coupled plasma spectrophotometry (ICP), IR spectroscopy, SEM, and HRTEM. The results verified that the size and morphology of Fe₂O₃/AP nanocomposites are controllable, and the dispersion of Fe₂O₃ nanoparticles is greatly improved in Fe₂O₃/AP nanocomposites. Moreover, the thermal decomposition of the as‐prepared Fe₂O₃/AP nanocomposites was measured with TG‐DSC. The results showed that the Fe₂O₃ nanoparticles in Fe₂O₃/AP nanocomposites exhibit better catalytic activity on the thermal decomposition of AP. In addition, the mechanism was also discussed.     

逆向热致相分离法(RTIPS)制备PES/DMAc/DEG体系微孔膜及其性能表征

以PES/DMAc/DEG低临界共溶温度(LCST)体系为铸膜液,利用低临界共溶温度(LCST)的热致相分离(LCST-TIPS,简称RTIPS)法制备PES微孔膜.探究影响PES微孔膜理化性能及其结构的2个主要因素:凝胶浴温度、非溶剂(DEG)/溶剂(DMAc)的质量比.运用扫描电镜(SEM)﹑纯水通量﹑BSA截留率...     

Synthesis,In Vitro,and Computational Studies of PTP1B Phosphatase Inhibitors Based on Oxovanadium(IV) and Dioxovanadium(V) Complexes

One of the main goals of recent bioinorganic chemistry studies has been to design and synthesize novel substances to treat human diseases. The promising compounds are metal-based and metal ion binding components such as vanadium-based compounds. The potential anticancer action of vanadium-based compounds is one of area of investigation in this field. In this study, we present five oxovanadium(IV) and dioxovanadium(V) complexes as potential PTP1B inhibitors with anticancer activity against the MCF-7 breast cancer cell line, the triple negative MDA-MB-231 breast cancer cell line, and the human keratinocyte HaCaT cell line. We observed that all tested compounds were effective inhibitors of PTP1B, which correlates with anticancer activity. [VO(dipic)(dmbipy)]·2 H₂O (Compound 4) and [VOO(dipic)](2-phepyH)·H₂O (Compound 5) possessed the greatest inhibitory effect, with IC₅₀ 185.4 ± 9.8 and 167.2 ± 8.0 nM, respectively. To obtain a better understanding of the relationship between the structure of the examined compounds and their activity, we performed a computer simulation of their binding inside the active site of PTP1B. We observed a stronger binding of complexes containing dipicolinic acid with PTP1B. Based on our simulations, we suggested that the studied complexes exert their activity by stabilizing the WPD-loop in an open position and limiting access to the P-loop.     

5,10,15,20-四(4-羟基苯基)卟啉的合成及光谱性质的研究

以吡咯、对甲氧基苯甲醛和吡啶盐酸盐为原料,采用两步法合成了5,10,15,20-四(4-羟基苯基)卟啉(THPP),反应总收率39.2%,产品纯度大于97%;采用微波加热法提高了该产物的合成效率;用1HNMR、HPLC-MS和HRMS对化合物进行了结构和纯度表征;研究了其在不同溶剂和不同pH值条件下的光谱性质.