Inhibited interleukin 35 expression and interleukin 35–induced regulatory T cells promote type II innate lymphoid cell response in allergic rhinitis

BackgroundInterleukin (IL)-35 and IL-35–producing regulatory T cells (iTr35) have been reported to inhibit T_H2 response in allergic rhinitis (AR). However, its effects on type II innate lymphoid cells (ILC2) are not well characterized.ObjectiveTo investigate the effect of IL-35 on ILC2 in AR.MethodsA total of 25 patients with AR and 20 controls were recruited. The expression and regulation of IL-35 receptor in ILC2 were analyzed by real-time polymerase chain reaction. The effect of IL-35 on ILC2 differentiation and cytokine production was analyzed by real-time polymerase chain reaction and enzyme-linked immunosorbent assay. In addition, iTr35 were cocultured with ILC2 to explore the effect of iTr35 on ILC2. The AR mice models were also established to confirm the role of IL-35 in vivo.ResultsThe patients with AR had decreased IL-35 expression and iTr35 proportion and increased ILC2 and type II cytokines compared with the controls. Notably, IL-35 inhibited ILC2 differentiation and type II cytokine production by regulating IL-12Rβ2 and gp130. IL-35 promoted the inducible costimulatory molecule expression by iTr35 and the inducible costimulatory molecule ligand expression by ILC2. IL-35–treated mice with AR presented decreased frequency and function of nasal ILC2.ConclusionIL-35 inhibited ILC2 responses directly or through mutual contact between iTr35 and ILC2 in AR, suggesting that IL-35 may be used as a potential treatment target in AR.     

Rapid characterization of the chemical constituents of Sanhua decoction by UHPLC coupled with Fourier transform ion cyclotron resonance mass spectrometry

Sanhua decoction, a famous Chinese herbal formula has been widely used for the treatment of stroke. In our study, a rapid, swift and straightforward analytical method with the help of UHPLC-FT-ICR-MS/MS was successfully developed for the first time to separate and identify the chemical constituents of Sanhua decoction. Chromatography was performed on a Universal XB C₁₈ column (150 mm × 2.1 mm, 1.8 μm) using a mobile phase containing 0.1% formic acid–water (A) and acetonitrile (B). A total of 137 compounds in Sanhua decoction were identified or tentatively characterized. The findings revealed the fact that Sanhua decoction mainly contains flavonoids (in Aurantii fructus immaturus and Rheum palmatum L.), anthraquinones (in Rheum palmatum L.), coumarins (in Notopterygii Rhizoma Et Radix), phenylpropanoid glycosides, alkaloids and lignans (in Magnoliae Officmalis Cortex), which made up the key ingredients existing in Sanhua decoction. This study is hoped to be meaningful for the characterization of components in other traditional Chinese medicines, and lay the foundation for research on the pharmacology of Sanhua decoction.

Sanhua decoction, a famous Chinese herbal formula has been widely used for the treatment of stroke.     

双向调控Cox-2表达对人食管癌EC9706细胞裸鼠移植瘤放射敏感性的影响

目的 探讨上调和下调Cox-2基因表达对食管癌EC9706细胞裸鼠移植瘤放射敏感性的影响。方法 构建针对Cox-2基因的siRNA载体及Cox-2基因真核表达载体,通过脂质体转染技术将其转入食管癌EC9706细胞,G418筛选得到稳定转染细胞系。荧光定量RT-PCR和Western blot分别检测细胞中Cox-2 mRNA和Cox-2蛋白表达水平;裸鼠移植瘤实验检测上调和下调Cox-2表达联合X射线照射对食管癌的生长抑制作用。结果 Cox-2下调组食管癌EC9706细胞内Cox-2基因表达明显降低,Cox-2上调组明显升高。Cox-2下调组裸鼠移植瘤平均体积明显小于对照组(F=34.26,P<0.05);Cox-2上调组的瘤体平均体积大于对照组(F=26.38,P<0.05)。20 Gy γ射线照射后Cox-2下调组瘤体平均体积较照射前明显减小(F=16.35,P<0.05);而上调组裸鼠皮下种植瘤平均体积较照射前无明显变化。结论 下调Cox-2表达能抑制人食管癌EC9706细胞裸鼠移植瘤的生长,增强瘤体的放射敏感性,上调Cox-2表达则使肿瘤辐射抗拒。     

锶盐对高浓度地塞米松作用下大鼠成骨细胞的影响

目的 探讨雷奈酸锶(Sr)对高浓度地塞米松(Dex)作用下大鼠成骨细胞（OB)的影响。方法 采用大鼠骨髓基质干细胞(BMSCs)来源的成骨细胞,运用MTT试剂盒和AKP试剂盒分别测定Sr对细胞增殖和分化的影响,定量PCR方法测定Sr对高浓度地塞米松作用下成骨钙素（OC)和转化生长因子-β(TGF-β)表达的影响。结果 Sr浓度在10-5、10-4 mol/L时,能够促进OB增殖和分化(P < 0. 01); Sr浓度为10-4mmol/L能够有效拮抗高浓度Dex( 10-5 mol/L)对成骨细胞分化抑制作用（P <0.01),并能逆转Dex对成骨相关基因OC和TGF-β的抑制作用(P <0. 05,P <0. 01)。结论 Sr能够对大鼠BMSCs来源的成骨细胞的增殖、分化有促进作用,同时10-4 mol/L Sr可以拮抗Dex( 10-5 mol/L)对ALP和成骨相关基因的抑制作用。为Sr防治糖皮质激素性骨质疏松提供了细胞学依据。     

三七止血作用的实验研究

目的评价三七的止血作用。方法采用肝脏局部创伤的方法复制小鼠肝脏局部创伤出血模型,实验分为空白组、羧甲基纤维素钠组、淀粉组、云南白药组和三七粉组,各组局部给药,观察药物对出血量的影响;灌胃给予大鼠三七粉,连续8 d后,静脉注射肝素300 U/kg,15 min后,参照小鼠方法复制大鼠肝脏局部创伤出血模型,测定出血量;灌胃给予大鼠三七粉等药物,连续8 d,舌下静脉注射肝素造成肝素化大鼠模型,测其活化部分凝血酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间(TT)。结果三七粉局部给药能减少小鼠肝脏局部创伤出血模型的出血量(P<0.05),但与羧甲基纤维素钠组、淀粉组、云南白药组比较无明显差异,提示三七粉局部给药能减少出血量的作用,可能与其细粉状态有关,属非特异性止血。三七粉灌胃能减少肝素化大鼠肝脏局部创伤模型的出血量(P<0.05),抑制率为16.1%,表明止血作用较弱。三七粉灌胃能缩短肝素化大鼠APTT、TT(P<0.05),抑制率分别为25.7%、18.8%,对PT无影响。提示,三七有止血作用,可能与干预内源性凝血途径,促纤维蛋白形成有关。结论三七粉体内外均有止血作用,其中体外止血可能是物理性、非特异性的,体内止血可能与其部分对抗肝素的抗凝作用有关。     

补肾解毒方对放射致小鼠胸腺损伤的防护作用研究

背景　胸腺是最重要的中枢免疫器官,在接受射线后易受损造成免疫损伤,目前国内外研制的胸腺防护剂仍存在缺陷,中药因其高效低毒的特性,越来越受到重视。目的　研究补肾解毒方对放射致小鼠胸腺损伤的防护作用。方法　2019年1月,将90只C57BL/10J小鼠采用随机数字表法分为5组：空白对照组、单纯放射组、放射+氨磷汀组、放射+补肾解毒方防治结合组和放射+补肾解毒方预防组,每组18只。适应性饲养3 d后,放射+补肾解毒方防治结合组、放射+补肾解毒方预防组每日予以补肾解毒方汤剂灌胃（0.25 ml/只）,同时单纯放射组和放射+氨磷汀组予以等剂量0.9%氯化钠溶液连续灌胃10 d,第11天除空白对照组外,其余4组小鼠均接受与照射源同等距离的一次性4 Gy 60Coγ-射线全身照射,其中放射+氨磷汀组小鼠在照射前30 min于左侧腹腔内注射氨磷汀200 mg/kg。照射后,放射+补肾解毒方防治结合组继续予以补肾解毒方灌胃,余4组给予等剂量0.9%氯化钠溶液灌胃。观察小鼠一般情况,在照射后第1、7、14天进行取材,计算胸腺指数,在光镜下观察照射后第7天各组胸腺组织结构。结果　五组小鼠在照射后一般情况均较照射前差,具体表现为活跃度下降、精神状态不佳、皮毛光泽度下降,尤其单纯放射组程度更甚,其中5只小鼠出现毛发块状脱落症状。照射后第1、7、14天,空白对照组胸腺指数均高于其余4组（P<0.01）;照射后第7天,放射+氨磷汀组、放射+补肾解毒方防治结合组胸腺指数高于单纯放射组,放射+补肾解毒方预防组低于放射+补肾解毒方防治结合组（P<0.01）;照射后第14天,放射+补肾解毒方防治结合组胸腺指数高于单纯放射组、放射+氨磷汀组和放射+补肾解毒方预防组（P<0.05）。空白对照组胸腺结构完整,皮质内可见髓质,中央可见圆形或椭圆形胸腺小体;单纯放射组小梁结构破坏,界限不清,胸腺小体模糊;其余3组病理表现基本一致,部分小梁小叶结构破坏,胸腺小体隐约可见,皮质髓质界限不清,其中放射+补肾解毒方预防组还表现出更多炎性细胞浸润。结论　补肾解毒方能减轻放射所致小鼠胸腺组织损伤,发挥了良好的放射防护作用。     

Synthesis and photophysical studies of self-assembled multicomponent supramolecular coordination prisms bearing porphyrin faces

Multicomponent self-assembly, wherein two unique donor precursors are combined with a single metal acceptor instead of the more common two-component assembly, can be achieved by selecting Lewis-basic sites and metal nodes that select for heteroligated coordination spheres. Platinum(II) ions show a thermodynamic preference for mixed pyridyl/carboxylate coordination environments and are thus suitable for such designs. The use of three or more unique building blocks increases the structural complexity of supramolecules. Herein, we describe the synthesis and characterization of rectangular prismatic supramolecular coordination complexes (SCCs) with two faces occupied by porphyrin molecules, motivated by the search for new multichromophore complexes with promising light-harvesting properties. These prisms are obtained from the self-assembly of a 90° Pt(II) acceptor with a meso-substituted tetrapyridylporphyrin (TPyP) and dicarboxylate ligands. The generality of this self-assembly reaction is demonstrated using five dicarboxylate ligands, two based on a rigid central phenyl ring and three alkyl-spaced variants, to form a total of five free-base and five Zn-metallated porphyrin prisms. All 10 SCCs are characterized by ³¹P and ¹H multinuclear NMR spectroscopy and electrospray ionization mass spectrometry, confirming the structure of each self-assembly and the stoichiometry of formation. The photophysical properties of the resulting SCCs were investigated revealing that the absorption and emission properties of the free-base and metallated porphyrin prisms preserve the spectral features associated with free TPyP.The use of sunlight is ubiquitous as the input for carbon-neutral, renewable energy schemes (1). Every strategy that relies on solar energy conversion, ranging from direct conversion to electricity in photovoltaics (2) to the generation of fuels via electrocatalysis (3), photoanode (4, 5), or photocathode devices (6), or photocatalysis (7) requires that photons be absorbed by a molecule or material as the first step in providing the driving force for subsequent transformation. Natural systems have evolved light-harvesting complexes, comprising multiprotein ensembles embedded with pigment molecules to enhance photon absorption for photosynthesis (8). These pigment-rich sites are arranged such that excitation of a distal chromophore will ultimately result in energy transfer to a reaction center via a series of migration and transfer processes (9). Strategies to replicate natural light-harvesting complexes necessarily demand the organization of multiple chromophores (10), a requirement that makes self-assembly and supramolecular chemistry particularly well suited for such efforts (11–13). As such, a variety of approaches have been applied toward the development of new materials that exhibit broadband absorption and efficient energy transfer (14–16). The subsequent studies of such materials span investigations of the fundamental science behind energy migration and transfer, to practical applications in devices.Many of these designs incorporate porphyrin-based molecules (17–22), a pigment that is related to the ubiquitous chlorophyll found in natural light-harvesting systems (23). Porphyrins possess characteristic structural and photophysical properties that make them well suited for adaptation into artificial designs, not only for solar energy conversion but also for applications in photodynamic therapy (24–29), enzyme mimics (30–32), catalysis (33–37), and molecular electronic devices (38, 39). In the context of solar energy, many multiporphyrin light-harvesting designs have been developed ranging from organic polymers, metal-organic frameworks, and supramolecular ensembles. Porphyrins are remarkable precursors for incorporation into supramolecular designs due to their established syntheses that facilitate well-defined functionalization, the commercial availability of simple variants, and their ability to accommodate a wide range of metal ions, thus unlocking suites of complexes for a given design (40, 41). The absorption spectra of porphyrin molecules are dominated by so-called Soret bands in the 380- to 500-nm range, accompanied by a set of weaker, but still considerably intense Q bands in the 500- to 700-nm range. Because these bands result in the absorption of visible photons, the attractiveness of porphyrins in solar conversion schemes becomes apparent. Furthermore, the emission wavelengths of a given porphyrin may align with its Q-band absorption, providing the spectral overlap that is required for efficient energy migration.Although extended structures such as polymers and metal-organic frameworks can organize similar numbers of pigments as are found in natural systems (42–47), their study can be complemented by relatively smaller supramolecules that facilitate characterization and study, preserving attractive properties such as solubility and facile tenability (48, 49). As such, coordination-driven self-assembly has been used to explore porphyrin-based supramolecular coordination complexes in traditional one- or two-component schemes (50-53).The recent development of new strategies for multicomponent self-assembly has provided a method to increase the structural complexity of chromophoric supramolecular coordination complexes (SCCs) wherein three or more building blocks can assemble into a single thermodynamically favored product (54, 55). One method to achieve multicomponent assembly is to identify metal nodes and ligands that prefer heteroligated coordination spheres, thus avoiding statistical mixtures of products. For example, when Pt(II) ions are combined with pyridyl and carboxylate-based ligands, mixed Pt-N,O coordination environments are formed exclusively (56). This method has been used recently to construct a variety of 2D and 3D SCCs and also provides the basis for supramolecule-to-supramolecule transformation, wherein a Pt-pyridyl SCC may be mixed in the proper stoichiometry with a Pt-carboxylate SCC to quantitatively furnish a third multicomponent SCC (56). Furthermore, traditional two-component assembly typically requires the use of rigid building blocks so as to maintain the angularity and directionality that determines the structural outcome of a reaction. In contrast, multicomponent approaches also permit the use of structurally ambiguous ligands, provided the remaining building blocks enforce a proper degree of directionality. As such, alkyl-based building blocks may be used in such self-assembly schemes, despite the fact that rotation about the carbon–carbon bonds means that a single orientation of the coordination vectors is not maintained (57).We report here a unification of many recent themes of coordination-driven self-assembly: the formation of multichromophore complexes as motivated by the need for new light-harvesting materials, multicomponent self assembly using Pt(II) heteroligation, and alkyl-based dicarboxylates in the formation of 3D prisms that demonstrate a hitherto-underexplored class of building blocks for coordination-driven self-assembly. The synthesis and characterization of a suite of 10 prisms is described, thus demonstrating the structural modularity and tunability that is possible through self-assembly approaches. Five prisms are formed containing free-based tetrapyridyl porphyrin, with the remaining five being their Zn-containing analogs. The absorption and emission properties of the SCCs are evaluated and compared with free tetrapyridylporphyrin (TPyP) revealing that, upon incorporation into prisms, the photophysical properties remain largely unchanged. In addition, the solubility of the prisms far exceeds that of the parent TPyP, indicating that coordination-driven self-assembly may be a powerful method to optimize the physiochemical characteristics of new light-harvesting materials.     

凋亡调节蛋白Fas和Caspase-3在乳腺癌中的表达及意义

目的观察人乳腺癌组织中凋亡调节蛋白Fas和Caspase-3的表达,探讨Fas和Caspase-3与乳腺癌发生、发展的关系。方法收集手术切除的人乳腺癌组织和癌旁相对正常乳腺组织(距癌组织5 cm以上),用免疫组织化学方法对23例乳腺癌标本进行检测。结果在相对正常乳腺组织和乳腺癌组织中Fas蛋白的阳性表达率分别为86.9%和47.3%;乳腺癌组织中Fas的阳性表达率明显低于相对正常乳腺组织。在相对正常乳腺组织和乳腺癌组织中Caspase-3的阳性表达率分别为82.6%和43.5%;乳腺癌组织中Caspase-3的阳性表达率明显低于相对正常乳腺组织。乳腺癌组织中caspase-3与Fas的表达率均与淋巴结转移无关。相关性分析表明,乳腺癌癌中的表达caspase-3与Fas表达呈正相关。结论乳腺癌组织中Fas和Caspase-3表达下调,使肿瘤抗凋亡机制过度激活,可能在乳腺癌的发生和发展过程中起重要作用。     

有晶状体眼后房型人工晶状体植入术后前房角的改变及相关因素分析

目的：探讨有晶状体眼后房型人工晶状体植入术后不同时间的前房角变化,分析影响术后前房角度数（ACA）的相关因素。方法：前瞻性非随机的连续病例研究。连续收集2018年9月至2019年2月在郑州大学第一附属医院眼科屈光中心接受有孔型有晶状体眼后房型人工晶状体（ICL）植入术的近视患者81例（81眼）,患者术前,术后1 d、1个月和6个月行Visante眼前段光学相干断层扫描（OCT）检 查,测量术前、术后鼻侧和颞侧ACA及术后拱高。分析术后ACA与术前等效球镜度（SE）、眼轴长度、水平角膜直径、前房深度、前房容积、水平角膜曲率、术前ACA、ICL直径和术后拱高的相关性。 采用重复测量方差分析比较患者不同时间点眼前段各项参数的变化,两两比较采用 Bonferroni检验。 相关性分析采用Pearson相关或Spearman相关分析。结果：术后1 d、1个月和6个月的ACA比较差异 均无统计学意义,术后的鼻侧ACA较术前减少45%～50%（F=268.73,P<0.001）,术后的颞侧ACA 较术前减少44%～49%（F=333.40,P<0.001）。ICL植入术后1 d、1个月和6个月的平均拱高分别为 （640±180）μm、（540±150）μm和（530±160）μm,差异具有统计学意义（F=54.14,P<0.001）;术后 1个月内拱高下降趋势明显,术后1个月至术后6个月期间拱高下降趋势明显减缓。 ICL植入术后鼻 侧ACA与术前前房容积、术前前房深度和术前ACA呈正相关（r=0.426,P=0.003;r=0.377,P=0.008; r=0.525,P<0.001）;与ICL直径、拱高、术前SE、眼轴长度、水平角膜曲率和水平角膜直径无相关性。 ICL植入术后颞侧ACA与术前前房容积、术前前房深度和术前ACA呈正相关（r=0.392,P=0.006; r=0.376,P=0.008;r=0.594,P<0.001）,与ICL直径、拱高、术前SE、眼轴长度、水平角膜曲率和水平角膜直径无相关性。结论：ICL植入术后前房角明显减小,随访期内患者前房角开放度数基本稳定, 没有进一步变窄。