The compound Perillaldehyde (PA) is a bioactive constituent found in natural Perilla oil, exhibiting remarkable antibacterial and antioxidant activity. However, the instability of this compound caused by the easy oxidation of its aldehyde and olefin groups imposes limitations on its extensive applications. Therefore, it is crucial to develop effective strategies for enhancing the stability of PA. This study focuses on utilizing a series of natural cyclodextrins (CDs) to synthesize inclusion complexes (ICs) with PA, aiming to improve water solubility and achieve exceptional stability at room temperature, which is essential for their potential applications in biomedicine or food industries. The hydrophobic cavity of CDs can accommodate the hexatomic ring and hydrophobic chains of PA based on the well-known size-matching effect and hydrophobic interaction, thereby forming stable ICs. Additionally, the hydrophilic outer wall of CDs imparts excellent water solubility to ICs. Phase solubility investigations demonstrate successful construction of α-CD-PA IC and β-CD-PA IC with an inclusion ratio of 1:1. Their respective stability constant (KC) are determined as 342 L/mol and 180 L/mol, indicating superior stability for α-CD-PA IC compared to β-CD-PA IC. Conversely, γ-CD with a larger cavity fails to form a stable inclusion complex with PA due to inadequate size matching. Nuclear Magnetic Resonance Hydrogen Spectroscopy (1H and 2D NMR) studies reveal that PA enters the CDs cavity (α-CD or β-CD) from its wide rim with almost the entire molecule being obliquely embedded within it. Thermogravimetry Analysis (TGA) confirm that after inclusion with CDs, PA exhibits expected stability at 25 ℃. Furthermore, CDs-PA ICs demonstrate significantly improved water solubility compared to pure PA along with enhanced antioxidant activity and slow-release performance, rendering them highly favorable for various applications.

本研究目的在于使用哺乳动物真核表达系统获得具有高免疫原性重组人Jagged2胞外区C2-EGF2区段(aa27～309),为后续抗Jagged2抗体相关研究奠定基础.首先,合成JAG2胞外区基因并插入pcDNA3.1(+)质粒构建哺乳细胞真核表达载体.使用阳离子脂质体转染试剂瞬时转染HEK293F细胞,收集培养上清,使用镍离子亲和层析柱梯度洗脱纯化,纯化后蛋白样品经过12％胶浓度SDS-PAGE电泳,免疫Balb/c小鼠并利用间接ELISA法测试小鼠抗血清效价.结果显示,150 mmol/L咪唑浓度下洗脱出的Jagged2蛋白在相对分子质量35 000左右出现单一电泳条带,ImageJ分析纯度达90％以上.3次免疫后小鼠产生的抗血清效价最高可达106以上.此结果表明真核表达得到的Jagged2胞外区蛋白具有较高的纯度和免疫原性,可以作为免疫原蛋白支持后续的抗体筛选.

A novel fiber sensor which can detect the concentration of copper ions indirectly was proposed and demonstrated by coating the knotted-shaped fiber with chitosan. The diameter of the knotted-shaped fiber was determined to be 10 mm. The Mach-Zehnder Interference generated due to bending deformation that excited some core-modes into higher order cladding-modes and two modes interfered. Chitosan was selected as the identification material for coating the fiber. In order to enhance the chelation between chitosan and copper ions, and improve the sensitivity of copper ions, glutaraldehyde was used to chemically modify chitosan. The original fiber sensor was annealed to obtain a stable structure, and then it was immersed in the modified chitosan solution and deionized water to form an available chitosan coating on the surface of the sensor. In the detection experiment with copper ions concentration of 0-330 ppm, the results show that interference spectra uniformly red-shift with the increase of the copper ions concentration and the sensitivity of the copper ion was 24.6 pm/10ppm. At the same time, ideal spectral fluctuations were demonstrated in the stability inquiry experiments.

目的:利用傅里叶红外变换光谱对干燥综合症伴龋齿患者口腔内特征微生物进行快速鉴别.方法:选取白色念珠菌、发酵乳杆菌、变异链球菌、干燥奈瑟菌和短双歧杆菌作为标准菌株与20例干燥综合症同时伴有龋病的患者口中分离的疑似菌株进行比对,采用傅里叶红外变换光谱法与标准菌株进行红外光谱的采集,光谱经过基线校正、光谱平滑和归一化处理,并采用一阶导数聚类分析.结果:分离的5种疑似菌与5种标准菌在特征谱区能准确的各自聚为一类.结论:利用傅里叶红外变换光谱能够快速、简便的分析干燥综合症患者口腔中的目标菌.

Methamphetamine (METH) is a highly neurotoxic psychoactive substance that can directly damage the central nervous system through prolonged use. Oxytocin (OT) has attracted much attention because of its neuroprotective effect. The purpose of this study was to investigate whether OT is neuroprotective against METH-induced damage in rat hippocampal neurons. Our results revealed that pre-incubation with OT significantly prevented the damage of METH to hippocampal neurons, including the decrease of mitochondrial membrane potential and the increase of ROS (reactive oxygen species). OT pre-incubation attenuated the up-regulation of Cleaved-Caspase-3 expression and the down-regulation of Bcl-2/Bax expression induced by METH. Pre-incubation with OT prevented the decrease in oxytocin receptor density and P-CREB (phosphorylation of cAMP-response element binding) expression induced by METH in rat hippocampal neurons. Moreover, Pre-incubation of atosiban (ATO) significantly prevented these changes. In conclusion, our study proved that pre-administration of OT could significantly attenuate hippocampal neuron apoptosis induced by METH. Oxytocin receptor activation is involved in the preventive effect of OT on METH-induced apoptosis in rat hippocampal neurons.

In this study, a new molecularly imprinted material, MIP@UiO-66-NH2, was synthesized with glutathione (GSH) as template and mesoporous metal organic framework (UiO-66-NH2) as matrix. The molecularly imprinted polymer was modified on the surface and into the pores of the UiO-66-NH2 by surface molecular imprinting method with thin polymer layer. Based on high specific surface area (1091.93 m(2) g(-1)) and appropriate pore size (35 nm) of the ordered mesoporous UiO-66-NH2, the adsorption capacity for GSH reached 94.43 mg g(-1), and the adsorption equilibrium could be achieved within 30 min. The adsorption isotherm data of MIP@UiO-66-NH2 could be described well by Freundlich model and the kinetic data complied well with pseudo-second-order model. In addition, the MIP@UiO-66-NH2 showed low adsorption capacity to GSH structural analogs (Q(L-cys) = 6.51 mg g(-1)), suggesting great selectivity for GSH recognition. Finally, the MIP@UiO-66-NH2 was successfully applied for selective separation of GSH from BSA, skim milk and egg white tryptic digest.

探讨单链抗体JZC00和糖酵解抑制剂2-脱氧葡萄糖(2-deoxyglucose,2-DG)的联合用药对小鼠非小细胞肺癌细胞LLC、小鼠乳腺癌细胞4T1的抗肿瘤作用.利用大肠埃希菌表达并纯化单链抗体,用SDS-PAGE和Western blot法鉴定JZC00;MTT法分析JZC00/2-DG联用组对肿瘤细胞体外增殖的抑制作用;葡萄糖和乳酸测定试剂盒测定培养基中葡萄糖和乳酸浓度,并计算肿瘤细胞葡萄糖摄取抑制率及乳酸释放抑制率;给药周期为15 d,给药同周期内测量肿瘤质量及体积.结果显示:经原核表达的JZC00相对分子质量正确,在体外能够抑制肿瘤细胞的增殖;JZC00及2-DG单独给药均能够抑制肿瘤细胞糖酵解,且JZC00/2-DG联合使用能够协同抑制糖酵解,当在体外模拟缺氧环境时JZC00抑制作用下降,加入2-DG后能够逆转其对糖酵解的抑制作用;两组体内模型显示与JZC00单药组相比,联合用药组抑瘤作用显著提高,2-DG能够提高抗血管生成抗体抑瘤效果,提示其联合在治疗实体瘤方面具有潜在价值.

细胞因子在免疫系统中发挥重要的调节作用.近年来应用细胞因子治疗肿瘤的临床试验大量展开,但细胞因子体内半衰期短、会产生严重的剂量限制性毒性,这大大限制了其在临床上的应用.随着抗体技术的发展,抗体-细胞因子融合蛋白(也称免疫细胞因子)通过抗体向病变部位靶向递送细胞因子,是一个解决细胞因子临床难题的可选方法.本文以白介素-2(Interleukin-2,IL-2)-免疫细胞因子为例,追踪近期治疗复发/难治/转移性肿瘤的临床试验,综述免疫细胞因子的结构特性,与细胞因子相比的优越性及与其他治疗方式的联用效果,旨在为免疫细胞因子进一步发展提供参考.

肿瘤转移是导致癌症患者死亡的主要原因.研究发现肿瘤发生转移前会在靶器官建立转移前生态位,为转移的肿瘤细胞提供适合和支持其定植的微环境.转移前生态位的建立涉及信号的传递和响应.原发肿瘤释放信号分子,直接或间接地改变靶器官部位细胞的行为,进而细胞分泌黏附因子、炎性因子、基质金属蛋白酶等,最终形成利于转移细胞定植的生态位.从细胞响应信号发生转变的角度,概述转移前生态位的建立过程,重点概括髓源性抑制细胞、中性粒细胞、巨噬细胞、驻留细胞及基质细胞响应信号分子发生转变的过程及相关机制,并探讨这一细胞转变过程在抗转移药物开发方面的应用价值.

OBJECTIVES:Alcohol consumption is a risk factor for stroke. However, there are no available data on the effect of alcohol consumption on the long-term outcome of ischemic stroke in China. Therefore, this study aimed to explore the association of alcohol consumption with the prognosis of ischemic stroke by subtype in different follow-up periods after stroke. METHODS:This 12-month follow-up study recruited 3830 acute ischemic stroke patients from Tianjin, China, between 2016 and 2018. Patients were categorized into two groups according to their consumption of alcohol. Differences in mortality, recurrence, and dependency rates at 3 and 12 months after stroke were compared between both groups. RESULTS:The mortality, recurrence, and dependency rates at 12 months after stroke were significantly higher in patients who previously consumed alcohol than in those without previous alcohol consumption (all P < 0.005). A similar trend was observed for mortality rate at 3 months after stroke (P < 0.001). The risk of death at 3 months after an atherothrombotic stroke decreased by 63.4% (relative risk [RR], 0.366; 95% confidence interval [CI], 0.144-0.935) among patients who previously consumed alcohol compared with those who never consumed alcohol. Moreover, for patients with small artery disease classified according to the Trial of ORG 10,172 in Acute Stroke Treatment (TOAST), the recurrence and dependency rates at 12 months after stroke decreased by 49.2% (RR, 0.508; 95% CI, 0.259-0.996) and 49.5% (RR, 0.505; 95% CI, 0.258-0.990), respectively, among patients who consumed alcohol. CONCLUSIONS:Previous alcohol consumption decreased the risk of death at 3 months after stroke among patients with atherothrombotic stroke according to the TOAST classification. Furthermore, for patients with small artery disease (according to TOAST classification), alcohol consumption significantly decreased the risk of recurrence and dependency at 12 months after stroke. This study highlights an urgent need to quantify the association of alcohol consumption with outcomes after stroke in China to improve stroke prognosis.

恶性肿瘤是全球范围内严重危害人类健康的疾病,并且发病率逐年上升.血管生成是肿瘤发生发展最重要的过程之一,90％的实体瘤都依赖功能性的血管网络提供氧气和营养.血管生成主要由血管内皮生长因子(VEGFs)和血管内皮生长因子受体(VEGFRs)调节,而通过单克隆抗体阻断VEGF/VEGFR2相互作用能够显著抑制肿瘤生长,是一种非常有前景的抗肿瘤策略.雷莫芦作为目前惟一一款上市的靶向VEGFR2的抗体药物,在多种癌症中取得了良好的临床效果.然而单靶点的治疗方法易产生耐药性及不良反应,多功能、多靶点的治疗策略如联合用药等成为目前肿瘤治疗的主流.同时,VEGFR2由于其在肿瘤细胞及肿瘤血管高表达的特性也成为了双特异性抗体良好的靶标.因此文章就靶向VEGFR2抗体的研究现状、挑战及以其为基础的多靶点治疗策略进行综述,并对肿瘤治疗的未来发展趋势进行展望.

心脏毒性是导致药物研发中断或撤市的主要原因之一.目前,临床前药物心脏安全性评价主要检测药物对hERG(Human ether-a-go-go)单一钾离子通道的阻断作用和药物对动物心电图QT间期的影响.这些检测假阳性率高,已经影响了新药研发的进程.人诱导多功能干细胞分化的心肌细胞(Human induced pluripotentstem cell-derived cardiomyocytes,hiPSC-CMs)具有人心肌细胞相似的结构与性质,为临床前药物心脏安全性评价提供了新的细胞模型,已经成为了未来心律失常检测方法——体外综合性心律失常检测(Comprehensive in vitro proarrhythmia assay,CiPA)的组成内容之一.此外这种技术也促进了非心律失常心脏毒性(包括药物导致的结构性心脏毒性与收缩性心脏毒性)检测方法的发展.文章介绍hiPSC-CMs的基本特性、在临床前药物心脏安全性评价中的研究进展以及局限性.

Reg蛋白家族是一种具有促进多种组织和细胞增殖、抑制凋亡等多功能的蛋白超家族.最新研究表明,Reg蛋白与多种疾病的发生、发展和治疗密切相关.重点阐述Reg蛋白在糖尿病、自身免疫疾病、炎症、癌症等疾病中发挥的作用及其作为新型药物应用于相关疾病治疗的研究进展,为Reg蛋白的药物研发提供了思路.

B细胞表面成熟抗原(B-cell maturation antigen,BCMA)是一种浆细胞选择性的蛋白,最早发现于成熟的B淋巴细胞表面,在其他组织细胞中几乎不表达.其在恶性增殖的B淋巴细胞(例如骨髓瘤细胞、白血病细胞)中高度表达,同时其介导的下游信号通路,对细胞的存活、增殖、转移和耐药中起着关键性的作用,这些特性使得它成为免疫治疗多发性骨髓瘤的一个靶点.近年来,针对BCMA的新型肿瘤免疫治疗方法日趋成熟,主要包括CAR-T(Chimeric Antigen Receptor T-Cell Immunotherapy)疗法、双特异性抗体(Bispecific antibody,BsAb)和抗体药物偶联物(Antibody-drug coupling,ADC)3大阵营,且目前已有4类药物进入临床Ⅰ期.从已经公开的临床前及临床Ⅰ期试验的结果来看,针对BCMA的靶向药物能够有效的靶向肿瘤细胞,并通过免疫细胞或小分子的细胞毒作用杀伤肿瘤细胞,药效显著,不良反应可控,是一个极具发展潜力的肿瘤靶向治疗新靶点.该文就BCMA的生物学功能、信号通路以及以其为靶点的肿瘤免疫治疗药物进行综述.

In this study, the exopolysaccharide from Bacillus amyloliquefaciens PB6 was purified and identified as levan. To explore the application of this levan in nanotechnology, its silver nanoparticles (AgNPs) were prepared in a simple and green method. Transmission electron microscopy analysis revealed that the size of the synthesised AgNPs was in the range of 19.47 ± 2.45 nm. X-ray powder diffraction analysis confirmed the face-centred cubic crystalline structure of metallic silver. Moreover, the AgNPs at the concentration of 0–12 μg/mL exhibited a significant inhibitive effect on Staphylococcus aureus and Escherichia coli. Tested by tetrazolium-based colourimetric assay, the viability of RAW 264.7 cells was higher than 75% in the presence of 20 μg/mL AgNPs. The results not only proved the feasibility of producing levan from B. amyloliquefaciens with high safety and a high yield, but also demonstrated the simple and cost-efficient approach of AgNPs synthesis as potential antibacterial agents from levan.

Colorectal carcinoma (CRC) is one of the most common malignant cancers worldwide. The poor response of CRC to chemotherapy has whipped up the interest in targeted therapy with monoclonal antibodies for its potential efficiency. However, cetuximab, as one of the first-line targeted drugs in the treatment of CRC, has drug resistance and poor prognosis in clinic. To address this, a novel bispecific protein with CRC targeting and natural killer (NK) cell triggering was used for treatment. NK cell-mediated immunosurveillance is normally activated by the activating receptor natural killer cell receptor NK group 2, member D (NKG2D), which binds its key ligand major histocompatibility complex (MHC) class I-related chain A (MICA) expressed on the tumor cells. To trigger NK cell-mediated cytotoxicity, we fused MICA portion to a single-chain antibody fragment rG7S targeting the tumor-associated antigen CD24. In vitro, flow cytometry, cytotoxicity assay, degranulation, and cytokines release assay revealed that the fusion protein rG7S-MICA could both binds to CD24 and NKG2D which enhances NK cell sensitivity and NKG2D-mediated immunosurveillance against CD24+ CRC cells. Furthermore, in a CD24+ CRC-bearing nude mice model, rG7S-MICA effectively recruits NK cell to the tumor site and increase the release of cytokines such as interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α), and shows potential antitumor effects. In conclusion, rG7S-MICA provides a novel immunotherapeutic strategy for CRC, which could be further developed against other CD24+ malignancies.

Cellular interactions between endothelial cell (EC) and vascular smooth muscle cell (VSMC)/macrophages seem to be greatly changed under inflammatory conditions. Although simvastatin could regulate inflammatory transcription factors in EC and VSMC and also could inhibit leukocyte-endothelium interaction, whether it could modulate VSMC/macrophage functions that are induced by tumor necrosis factor-alpha (TNF-alpha)-activated EC remained unclear. The purpose of this study was to investigate the effects of simvastatin on VSMC/macrophage functions, which are induced by TNF-alpha-activated EC in coculture system in vitro. The results showed that under noncontacting conditions, simvastatin could reduce the proliferation, apoptosis, and TNF-alpha, IL-6, and vascular endothelial growth factor secretion both in VSMC and macrophage, which is induced by TNF-alpha-activated EC. And a hypothesis that simvastatin regulates the interactions and the soluble factors between EC and VSMC/macrophages could be drawn. And that might be a potential anti-atherosclerosis mechanism of simvastatin.

OBJECTIVE To isolate and purify an active polysaccharide of Momordica charantia L.and investigate its struc-tural characterization and immunomodulatory and antitumor activities.METHODS A polysaccharides was isolated from Mo-mordica charantia L.in hot water extraction followed by dialysis and purification through anion exchange cellulose chromatog-raphy and gel filtration chromatography,and its molecular weight was measured by high performance gel permeation chroma-tography.Additionally,Fourier transform infrared spectroscopy(FT-IR)and nuclear magnetic resonance(NMR)were used to determine its partial structural characterization,and its immunomodulatory and antitumor activities were measured by cell and molecular biological techniques and pharmacological methods.RESULTS A heteropolysaccharide named MCP-2 with molecu-lar weight of 745 kDa was isolated and purified,which was shown to be consisted of rhamnose,galacturonic acid,galactose, xylose,and arabinose with corresponding molar ratio being 1.1 9 ∶ 1 5.97 ∶ 3.40 ∶ 0.73 ∶ 0.94.Furthermore,MCP-2 was found to promote the lymphocyte proliferation of normal mice and to raise the levels of NO,IL-1β and TNF-α produced by macrophage RAW264.7.Intriguingly,MCP-2 presented the antitumor activity against S180 cells in vivo as well.CONCLU-SION All experimental data showed that MCP-2 had apparent immunologic enhancement and antitumor activities.

The gene fragment encoding acyl-CoA ligase DptE and its mutant DptE-296 were amplified using polymerase chain reaction(PCR) and were inserted into the prokaryotic expression vector pet22b through the restriction enzymes bamH Ⅰ and Xho L The recombinant plasmid was identified by screening positive clones using ampicillin,bacteria PCR and DNA sequencing.The correct plasmid was transformed into E.coliBL21 (DE3),induced by IPTG,extracting periplasmic proteins by osmotic shock method,purified by nickel affinity chromatography,identified by SDS-PAGE and Western blot.The purified protein was connected to biotin to assay the affinity with decanoic acid by fortebio.The recombinant vector DptE-pet22b and DptE-296-pet22b was successfully constructed.SDS-PAGE and Western blot showed that the expressed protein was consistent with the expected.Fortebio experiments showed that both DptE and DptE-296 can bind decanoic acid,but the combination is not strong.

Chinese hamster ovary (CHO) cells have become the mainstream in expression systems for producing therapeutic antibodies.The cultivation of CHO cells and expression of antibody are extremely related to culturing environment.We previously generated a fusion antibody named JZB01 targeting vascular endothelial growth factor receptor 2 (VEGFR2) and natural killer cells with significant anti-tumor activity but limited production.In this study,we aim to develop a robust method for fed-batch fermentation using design of experiments (DOE) with Minitabl 6.0,and evaluate the influence of commercial media,supplements and culture conditions on cell viability and antibody production.To improve the yield of fusion protein and establish a strategy for further study,the optimization of the production was undertook with orthogonal experimental analysis for four factors and the level was assessed by the response.Furthermore,in order to sustain the stability and uniformity of fusion protein for further research and diagnosis,the bioprocess was scaled up to a 3-L bio-reactor under the conditions of commercial medium CDM4PerMAb,supplemented with Boost2 + Boost5,12％ (V/V) volume,1 × 106 cells/mL of initial density and 2 days of feeding.The final yield was 54.45 mg/L and Western blot was adopted to identificated the affinity of fusion protein preliminarily,which was characterized as a homodimer with binding ability to each receptor.By the investigation for anti-angiogenesis function of JZB01 on HUVEC cells and anti-tumor function on MDA-MB-231 cells and K562 cells,the MrTT assay confirmed that the fusion protein had anti-angiogenic effect and enhanced anti-tumor activities.Thus,by establishing a platform to obtain fusion protein with high molecular weight in CHO cells system along with moderate costs,this study provides a robust methodology for producing therapeutic antibodies at laboratory level.And more trials will be taken to enlarge the application to industrial scale which could be applied in vitro and vivo preclinical tests.