一株鹅源禽腺病毒4型的分离及致病性

2023年,昆明一朗德鹅场部分雏鹅出现软脖子症状,伴有零星死亡,病变疑似禽心包积液综合征(hydropericardium syndrome,HPS)。本研究旨在对本次雏鹅病因进行分子病毒学诊断和病原的致病性研究。采集肝病料接种鸡肝癌细胞(leghorn male hepatocellular cells,LMH):1)观察细胞病变效应(cytopathic effect,CPE),取培养液负染、电镜观察病毒粒子形态;2)靶标4型禽腺病毒(serotype 4 fowl adenovirus,FAdV-4)的衣壳蛋白编码基因hexon基因进行PCR诊断,对扩增基因序列进行遗传演化分析;3)以5×10^(5.5)TCID_(50)/羽的剂量皮下注射感染10日龄健康泰州鹅,进行动物回归感染以确证其致病性。结果显示,将病料上清接种LMH细胞48~72 h后CPE明显,细胞圆缩、间隙变大、随后碎裂、逐渐脱落,电镜观察见二十面体无囊膜病毒粒子;PCR检测FAdV-4阳性,分离纯化病毒,命名为YNKM2023株;hexon基因的遗传演化分析显示,分离株与国内已有的鸡、鸭FAdV-4流行毒株相似性高达99.8%~99.9%,与印度分离株相似性为99.5%~99.6%。接种感染后,泰州鹅未出现典型的临床症状,但感染后3 d开始出现HPS的特征性病变,感染后6 d检测到中和抗体。综上,本研究成功分离到鹅源FAdV-4/YNKM2023株,该毒株与国内已有的鸡鸭流行毒株高度同源,尚未发生明显的基因漂移。分离株可以感染鹅,病毒可以在雏鹅体内复制引起组织器官病变并产生免疫应答,对鹅表现出一定的致病性,这有助于了解FAdV-4在鹅群的流行情况及其对雏鹅的致病性。

新型4-苯氧基吡啶类c-Met激酶抑制剂的合成和抗肿瘤活性研究

为了发现新型高活性c-Met激酶抑制剂,以2-氨基-4-氯吡啶为原料,经醚化、酰化、烷基化、还原和酰化等多步反应,制备了6种未见文献报道的4-苯氧基吡啶类化合物,化合物结构经红外光谱、核磁共振氢谱和质谱确证.生物活性测试结果显示,6种化合物对MKN-45、A549、H460和HT-294种肿瘤细胞株具有很好的抗增殖活性,两种化合物对c-Met激酶具有很好的抑制活性.其中化合物7b抑制MKN-45、A549、H460和HT-29细胞株的IC 50值分别为1.89、5.14、0.74、0.60μmol/L.该化合物在1000 nmol/L和200 nmol/L的浓度下对c-Met激酶的抑制率分别为96.2%和82.8%,显示出了优异的抗肿瘤活性.

NONHSAT248596.1内源性竞争miR-146a-5p调控骨关节炎软骨退变的机制

背景:目前已有针对lncRNA\miRNA\mRNA的共表达网络对骨关节炎发生发展调控机制的研究,课题组前期研究已通过数据库筛选出符合条件的NONHSAT248596.1和miR-146a-5p,尚缺乏体内实验来验证上述调控机制。目的:探究NONHSAT248596.1在基质细胞衍生因子1/4型趋化因子受体轴介导体内骨关节炎软骨退变进程中对miR-146a-5p发挥的竞争性内源性RNA调控作用。方法:取36只新西兰兔,通过向右侧后肢膝关节注射基质细胞衍生因子1溶液建立骨关节炎模型,采用随机数字表法分4组,lncRNA组、miRNA组、ceRNA组、对照组分别向造模膝关节内注射NONHSAT248596.1过表达的慢病毒载体、miR-146a-5p过表达的慢病毒载体、miR-146a-5p+NONHSAT248596.1过表达的慢病毒载体及空慢病毒载体。造模第4,8,12周,取膝关节软骨组织和软骨下骨组织进行相关检测。结果与结论:①苏木精-伊红与番红O固绿染色显示,4组软骨组织都有不同程度的退变表现,造模第4周时,lncRNA组软骨组织中的软骨细胞肿胀、细胞极性消失,细胞外基质破坏,出现表层糜烂、裂缝形成和软骨组织局部或全层缺失,并随时间延长软骨损伤程度逐渐加重,4组中miRNA组关节软骨炎症进展最缓慢;②qRT-PCR检测显示,相同时间点下,lncRNA组软骨组织中NONHSAT248596.1、4型趋化因子受体、基质金属蛋白酶3,9及13的mRNA表达量高于其他3组(P<0.05),miR-146a-5p、聚集蛋白聚糖及Ⅱ型胶原的mRNA表达量低于其他3组(P<0.05);造模后第8,12周,miRNA组软骨组织中的NONHSAT248596.1、4型趋化因子受体、基质金属蛋白酶3,9及13的mRNA表达量低于ceRNA组、对照组(P<0.05),miR-146a-5p、聚集蛋白聚糖及Ⅱ型胶原的mRNA表达量高于ceRNA组、对照组(P<0.05);③Western Blot检测显示,相同时间点下,lncRNA组软骨组织中的聚集蛋白聚糖及Ⅱ型胶原蛋白表达量始终低于其他3组(P<0.05);miRNA组造模后第8,12周软骨组织中的聚集蛋白聚糖及Ⅱ型胶原蛋白表达量高于ceRNA组、对照组(P<0.05);④结果表明,miR-146a-5p作为NONHSAT248596.1的作用靶点会受到其竞争性内源性RNA的作用造成活性被抑制,NONHSAT248596.1作用于miR-146a-5p后调控基质细胞衍生因子1/4型趋化因子受体轴,影响骨关节炎软骨组织中基质金属蛋白、Ⅱ型胶原、聚集蛋白聚糖的表达,造成细胞外基质的降解及蛋白多糖的丢失。

CMTM4调节宫颈癌髓源性抑制细胞分化和免疫效应

CMTM4蛋白异常表达与肿瘤发生、发展及预后相关,其在宫颈癌中的作用和机制尚不明确。本研究旨在探讨CMTM4在宫颈癌中的表达水平及其对MDSCs分化和免疫细胞功能的调节作用和分子机制。首先,利用免疫组化染色和Western印迹检测CMTM4在宫颈癌肿瘤和癌旁组织中的表达水平。结果显示,CMTM4在宫颈癌组织中的表达水平显著低于癌旁组织(P<0.01)。通过流式细胞术检测宫颈癌患者肿瘤样本中MDSCs的分化,使用CD45、CD11b和Ly6G标记M-MDSCs,使用CD45、CD11b和Ly6C标记G-MDSCs。发现宫颈癌患者血液中M-MDSCs和G-MDSCs的百分比显著高于健康志愿者组(P<0.01)。此外,流式分析发现,CMTM4高表达提高宫颈癌患者血液中CD3+CD4+T细胞和CD3+CD8+T细胞比例(P<0.01),促进Th1和Th17细胞的活化比例(P<0.05),抑制Th2和Treg细胞的活化比例(P<0.05或P<0.01)。通过流式分析肿瘤组织中CD8+PD-1+T细胞和CD8+PD-L1+T细胞的百分比。结果显示,CMTM4高表达抑制肿瘤组织中CD8+PD-1+T细胞和CD8+PD-L1+T细胞的百分比(P<0.05)。体外研究表明,CMTM4过表达的HeLa细胞显著抑制MDSCs生成(P<0.01),PD-1过表达的HeLa细胞进一步促进了MDSCs生成(P<0.05)。此外,使用ELISA检测Th1、Th2、Th17和Treg细胞因子的水平。发现CMTM4过表达的HeLa细胞可降低T细胞的IL-10分泌水平(P<0.01),而增加IFN-γ和TNF-α的分泌水平(P<0.01),其均被PD-1过表达逆转(P<0.05或P<0.01)。综上,本研究表明,CMTM4在宫颈癌中的低表达导致MDSCs的异常分化和肿瘤的免疫逃逸,CMTM4通过调节PD-1/PD-L1通路,从而影响肿瘤T细胞的活化。因此,CMTM4可能成为治疗宫颈癌的潜在靶点。

5-取代-4-氨基-3-巯基-1,2,4-三唑衍生物的应用进展

5-取代-4-氨基-3-巯基-1,2,4-三唑是一类重要的五元杂环化合物,由于分子中含有巯基和伯胺基两个活性基团,既可以单个基团参与反应生成N-或S-取代产物,又可以同时发生反应生成稠杂环化合物。由其衍生而来的化合物具有广泛的生物活性,在抗菌、抗肿瘤、抗结核、抗炎镇痛、酶抑制剂、荧光探针等方面都表现出优异的性能。综述了近年来5-取代-4-氨基-3-巯基-1,2,4-三唑衍生物在医学、农业及材料领域的应用,为今后进一步研究、开发此类化合物提供参考。

QSAR and Pharmacophore Studies of Thiazolidine-4-carboxylic Acid Derivatives as Novel Influenza Neuraminidase Inhibitors Using HQSAR, Topomer CoMFA and CoMSIA

In order to understand the chemical-biological interactions governing their activities toward neuraminidase （NA）, QSAR models of 28 thiazolidine-4-carboxylic acid derivatives with inhibitory influenza A virus were developed. The obtained HQSAR （hologram quantitative structure activity relationship）, Topomer CoMFA and CoMSIA （comparative molecular similarity indices analysis） models were robust and had good exterior predictive capabilities. Moreover, QSAR modeling results elucidated that hydrogen bonds highly contributed to the inhibitory activity, then electrostatic and hydrophobic factors. Squared multiple correlation coefficients （R2） of HQSAR, Topomer CoMFA and CoMSIA models were 0.994, 0.978 and 0.996, respectively. Squared cross-validated correlation coefficients （Q2） of HQSAR, Topomer CoMFA and CoMSIA models were in turn 0.951, 919 and 0.820. Furthermore, squared multiple correlation coefficients for the test set （R2test） of HQSAR, CoMFA and CoMSIA models were 0.879, 0.912 and 0.953, respectively. Squared cross-validated correlation coefficients for the test set （Q2ext） of HQSAR, Topomer CoMFA and CoMSIA models were 0.867, 0.884 and 0.899, correspondingly.

Synthesis of phenol-class azo derivatives of 4-aminosalicylic acid

To explore the better prodrug of 4-aminosalicylic acid（4-ASA）with higher activity and less side effects against the inflammatory bowel disease.4-ASA,after a succession of synthesis process,was conjugated with various carder molecules to get seven azo derivatives of 4-ASA.All compounds were characterized by FT-IR,^1H NMR,^13C NMR spectras in detail.New derivatives of 4-ASA were definituded.

Synthesis of azo derivatives of 4-aminosalicylic acid

For searching a better 4-aminosalicylic acid derivative with higher activity and less side effects against the inflammatory bowel disease, 4-aminosalicylic acid (4-ASA) was protected by benzyloxycarbonyl and acetyl, respectively. The resultant was hydrogenized to remove protective group of amino group, then the product was reacted with NaNO2 to give diazonium salt, which was conjugated with salicylic acid, hydroxybenzene, N-salicyloyl glycine acid to get azo derivatives of 4-ASA. The azo derivatives were hydrolyzed under the alkaline condition to get the target products. All compounds were characterized by FT-IR, (1)H NMR, (13)C NMR spectra in details. New derivatives of 4-ASA were characterized. The synthetic route was reasonable and feasible.

Non‑Magnetic Bimetallic MOF‑Derived Porous Carbon‑Wrapped TiO2/ ZrTiO4 Composites for Efficient Electromagnetic Wave Absorption

Modern communication technologies put forward higher requirements for electromagnetic wave(EMW)absorption materials.Metal-organic framework(MOF)derivatives have been widely concerned with its diverse advantages.To break the mindset of magneticderivative design,and make up the shortage of monometallic non-magnetic derivatives,we first try non-magnetic bimetallic MOFs derivatives to achieve efficient EMW absorption.The porous carbon-wrapped TiO2/ZrTiO4 composites derived from PCN-415(TiZr-MOFs)are qualified with a minimum reflection loss of−67.8 dB(2.16 mm,13.0 GHz),and a maximum effective absorption bandwidth of 5.9 GHz(2.70 mm).Through in-depth discussions,the synergy of enhanced interfacial polarization and other attenuation mechanisms in the composites is revealed.Therefore,this work confirms the huge potentials of nonmagnetic bimetallic MOFs derivatives in EMW absorption applications.

Synthesis of Calix[4] arene Thia Derivative and Extraction Effect of Substituents on Mercury( Ⅱ) and Lead( Ⅱ)

Calix[4]arene derivatives have been reported to possess high selectivity for metal ions.In order to analyze the extraction effect of substituents on Hg^(2+)and Pb^(2+),calix[4]arene derivatives containing hydroxyl,bromoethoxy and 1,11-dioxa-4,8-dithiahendecene were successfully synthesized,with their extraction effectiveness towards Hg^(2+)and Pb^(2+)were evaluated respectively.The results indicated that the phenolic hydroxyl in calix[4]arene improved the extraction ability on Hg^(2+)by promoting the formation of the negative oxygen ions that could bind with Hg^(2+)by coordination and ionic bonds.The extraction ability of 5,17-dinitro-26,28-(1’,11’-dioxa-4’,8’-dithiahendecene)-calix[4]arene(calix[4]arene thia derivative)was improved slightly due to the better coordination capacity for metal ions after introduction of 1,11-dioxa-4,8-dithiahendecene,which was a chelate-binding centre.Regarding to Pb^(2+),only 1,11-dioxa-4,8-dithiahendecene on molecule contributed to extraction due to the coordination bond.Since the metal ion with higher charge-to-radius ratio could coordinate with one ligand more stably,Pb^(2+)with higher charge-to-radius ratio than Hg^(2+)was coordinated and extracted by the calix[4]arene thia derivative more easily.Furthermore,extraction rates of the calix[4]arene thia derivative on both ions(Hg^(2+)and Pb^(2+))increased with the increase of pH value in acidic aqueous system(pH<7).

Synthesis, Crystal Structure and Biological Properties of One Zinc(Ⅱ) Complex Based on 4-Acyl Pyrazolone Derivative

[Zn（L）（CH3OH）3]（H2L = N-（1-phenyl-3-methyl-4-propenylidene-5-pyrazolone）-salicylidene hydrazide） has been synthesized by the reaction of zinc nitrate and ligand H2 L. The complex crystallizes in orthorhombic system, space group Pbca with a = 5.888（15）, b = 14.564（14）, c = 22.20（2） , V = 5137（8） 3, Z = 8 and F（000） = 2184. The ligand serves as a negative bivalent tridentate chelating agent to coordinate with the central zinc（II） atom. DNA-binding was studied by UV-Vis spectral analysis and ethidium bromide（EB） displacement experiments. The results showed that the DNA-binding constant of the complex is 5.1×104 M–1. Antitumor activity of [Zn（L）（CH3OH）3] and the ligand have been investigated by MTT assay, which indicated that the complex has better cytotoxicity to Eca-109 and He La than free ligand.

Synthesis of 4″-benzyloxyimino-4″-deoxyavermectin Bla derivatives

Six new 4＂-benzyloxyimino-4＂-deoxyavermectin B la derivatives were synthesized from avermectin Bla by the selective protection of C-5-hydroxy group, oxidation of C-4＂-hydroxy group, and deprotection followed by reaction with O-substituted hydroxylamine hydrochlorides. Their structures were confirmed by IR, 1H NMR, 13C NMR and MS. Insecticidal activities of the derivatives against Phopalosiphum pseudobrassicae, Spodoptera exigua and Pluteua xylosteua were evaluated.

Inhibition of Na^+,K^+-ATPase in Housefly (Musca domestica L.) by Terpinen-4-ol and Its Ester Derivatives

To reveal the insecticidal mechanism of terpinen-4-ol, the activity of Na＋,K＋-ATPase in insects tested were determined in vivo and in vitro. The results showed that terpinen-4-ol and its ester derivatives had strong contact activity to housefly and the contact toxicities of its derivatives except Z3 were all superior or equivalent to terpinen-4-ol. All the 7 compounds had strong inhibition towards activity of Na＋,K＋-ATPase. With poisoning symptom exacerbating, the inhibition rates were gradually increased. In vitro, the IC50 of terpinen-4-ol, Z1, Z2, Z4, Z5, and Z6 was 155.89, 197.98, 96.02, 121.36, 124.85, and 153.74 μg mL% respectively. There was well correlation between the LDs0 of terpinen-4-ol derivatives to housefly and the IC50 of terpinen-4-ol derivatives to Na＋,K＋-ATPase in housefly. In conclusion, Na＋,K＋-ATPase was likely the target of terpinen-4-ol against insects.

基于CoMFA方法对氟喹啉-4-酮衍生物的分子建模与设计

基于比较分子力场分析法(CoMFA),建立了16个已知活性的氟喹啉-4-酮衍生物抗肝癌活性(K_(S))的三维定量构效关系(3D-QSAR)模型,并研究该类结构与生物活性之间的关系.CoMFA模型的交叉验证系数(Q^(2))为0.338,拟合验证系数(R^(2))是0.987.此3D-QSAR模型的预测值与实验值基本一致,表明该模型具有显著的统计学可靠性和预测能力.该模型中立体场、静电场贡献率依次为41.9%、58.1%.根据3D-QSAR模型分析结果进行分子设计并完成活性预测,预测结果印证了分析的合理性,为该系列化合物的结构优化提供了合理建议.

Synthesis of thiourea derivatives as CCR4 antagonists

A series of thiourea derivatives have been synthesized. Their structures were confirmed by MS and 1H NMR. Several compounds showed potent activities as antagonists of CCR4 receptor.

Quantum Chemical Studies on the Corrosion Inhibition of Mild Steel by Piperidin-4-One Derivatives in 1 M H₃PO₄

The corrosion inhibition properties of 2,6-diphenylpiperidin-4-one (DPP) (1A) and 2,6-diphenyldihydro-2H-thiopyran-4(3H)-one (DPDT) (1B) for mild steel in 1 M phosphoric acid were studied using weight loss, potentiodynamic polarization and electrochemical impedance spectroscopic techniques. The effect of temperature on the corrosion behavior of mild steel has been examined in the temperature range 303 - 328 K. The inhibition efficiency increases with increasing inhibitor concentration but decreases with increasing temperature. Potentiodynamic polarization studies indicated the mixed nature of inhibitors. The adsorption of the inhibitors on mild steel surface obeyed the Langmuir adsorption isotherms. The density functional theory (DFT) at the B3LYP/6- 31G (d) basis set level was performed on 1A and 1B to investigate the correlation between molecular structure and the corresponding inhibition efficiency (%). The quantum chemical parameters such as EHOMO, ELUMO, the energy gap (E), hardness (η), softness (S), dipole moment (μ), electron affinity (A), ionization potential (I), the absolute electronegativity (χ), the fraction of electron transferred (N), electrophilicity index (ω), the back-donation (EBack-donation) and Mulliken population analysis have been calculated.

Use of Ultrasound and Microwave Irradiation for Clean and Efficient Synthesis of 3,3’-(Arylmethylene)bis (2-hydroxynaphthalene-1,4-dione) Derivatives

Nine 3,3’-(arylmethylene)bis(2-hydroxynaphthalene-1,4-dione) derivatives were synthesized through the reaction between 2-hydroxy-1,4-naphthalen-1,4-dione and different aromatic alde-hydes in water applying ultrasonic irradiation for 5 min at room temperature and microwave irradiation for 15 min at 70°;C. Two of the nine derivatives, compounds 3-e and 3-i, obtained from 3-bromo-hydroxybenzaldehyde and 5-methylfuran-2-carbaldehyde, respectively, are previously unpublished. The structures of all compounds were established on the basis of their spectral data and mass analysis. The attractive features of this synthesis protocol include mild conditions, high atom-economy and excellent yields with the elimination of water as the only by-product.

STUDIES ON THE SYNTHESIS AND BIOLOGICAL ACTIVITY OF 1-ARYLOXYACETYL-4-AROYLTHIOSEMICARBAZIDE DERIVATIVES

A series of new 1-aryloxyacetyl-4-aroylthiosemicarbazides were synthesized by means of solid-liguid phase transfer catalysis. The promoting effects of these new compounds on wheat growth were observed.

Synthesis and Characterization of Thiacalix[4] arene Derivatives with Polymerisable Groups

Introduction Calixarenes are versatile host molecules for molecular recognition and supramolecular assembly because its functional groups can be readily introduced into the phenolic OH or the para position to realize a wide variety of functions calixarenes-based polymers tion as these polymers can During the past decade, received increasing attenbe used to synthesize the materials that are suitable for the preparation of chemical sensor devices such as ion-selective electrodes or transport membranes.

Synthesis and activities of new 4-hydroxybenzoxazolone derivatives

A series of new 4-substituted benzoxazolone derivatives were synthesized according to a convenient method,their anti-inflammatory and analgesic activities in vivo were evaluated.All of them were new compounds,the structures of all the synthesized compounds were confirmed by ~1H NMR,^(13)C NMR,ESI-MS and HR-MS.Most of the compounds exhibited anti-inflammatory activity.