Direct regulation of IL-2 by curcumin

Interleukin-2 (IL-2) is a crucial growth factor for both regulatory and effector T cells. Thus, IL-2 plays a critical role in the stimulation and suppression of immune responses. Recently, anti-IL-2 antibodies (Abs) have been shown to possess strong IL-2 modulatory activities by affecting the interaction between IL-2 and IL-2 receptors. In this study, we screened an herbal library to identify a compound that regulates IL-2, which resulted in the identification of curcumin as a direct binder and inhibitor of IL-2. Curcumin is a phytochemical with well-known anti-cancer properties. In this study, curcumin mimicked or altered the binding pattern of anti-IL-2 Abs against IL-2 and remarkably inhibited the interaction of recombinant IL-2 with the IL-2 receptor α, CD25. Interestingly, curcumin neutralized the biological activities of IL-2 both in vitro and in vivo. In this report, we elucidated the unsolved mechanism of the anti-cancer effect of curcumin by identifying IL-2 as a direct molecular target. Curcumin, as a small molecule IL-2 modulator, has the potential to be used to treat IL-2 related pathologic conditions.     

甘氨鹅脱氧胆酸钠通过MAPK/ERK1/2介导Bcl-2在Ser70位点的磷酸化引起人肝细胞癌的抗药

B细胞淋巴瘤-2（Bcl-2）是一种重要的抗凋亡蛋白质,在多种人类肿瘤中普遍过表达。甘氨鹅脱氧胆酸钠（GCDA）与消化道肿瘤的发生发展密切相关,并能介导肝癌细胞对化疗药物的抵抗。本文旨在探讨在GCDA介导的人肝细胞癌（HCC）耐药性中Bcl-2的作用及其机制。本研究以肝癌细胞系为研究对象,Western印迹结果显示,Bcl-2在多种肝癌细胞系中均有表达。设计靶向Bcl-2的siRNA沉默HCC细胞系内源性Bcl-2的表达,发现Bcl-2沉默之后促进了化疗药物5-FU介导的HCC细胞凋亡。机制上,GCDA可介导Bcl-2在Ser70位点的磷酸化,而Ser70位点的磷酸化能够被PD98059（MAPK/ERK1/2抑制剂）所抑制。构建huBcl2-WT和huBcl2-S70A真核表达载体,脂质体转染HCC细胞系。用Annexin V-FITC/PI流式细胞术检测凋亡细胞。结果显示,huBcl2-WT过表达能抑制5 FU介导的凋亡,S70位点失活突变成A后,Bcl-2的过表达不能抑制5-FU介导的凋亡。本研究提示,GCDA通过MAPK/ERK1/2通路介导的Bcl-2 Ser70位点的磷酸化,在肝癌细胞的存活和抗药中发挥重要作用。抑制Bcl-2能够促进化疗药物5-FU介导的HCC细胞凋亡,该结果为治疗GCDA介导的耐药性肝癌提供新的思路。     

甘氨鹅脱氧胆酸钠通过MAPK/ERK1/2介导Bcl-2在Ser70位点的磷酸化引起人肝细胞癌的抗药

B细胞淋巴瘤-2（Bcl-2）是一种重要的抗凋亡蛋白质,在多种人类肿瘤中普遍过表达。甘氨鹅脱氧胆酸钠（GCDA）与消化道肿瘤的发生发展密切相关,并能介导肝癌细胞对化疗药物的抵抗。本文旨在探讨在GCDA介导的人肝细胞癌（HCC）耐药性中Bcl-2的作用及其机制。本研究以肝癌细胞系为研究对象,Western印迹结果显示,Bcl-2在多种肝癌细胞系中均有表达。设计靶向Bcl-2的siRNA沉默HCC细胞系内源性Bcl-2的表达,发现Bcl-2沉默之后促进了化疗药物5-FU介导的HCC细胞凋亡。机制上,GCDA可介导Bcl-2在Ser70位点的磷酸化,而Ser70位点的磷酸化能够被PD98059（MAPK/ERK1/2抑制剂）所抑制。构建huBcl2-WT和huBcl2-S70A真核表达载体,脂质体转染HCC细胞系。用Annexin V-FITC/PI流式细胞术检测凋亡细胞。结果显示,huBcl2-WT过表达能抑制5 FU介导的凋亡,S70位点失活突变成A后,Bcl-2的过表达不能抑制5-FU介导的凋亡。本研究提示,GCDA通过MAPK/ERK1/2通路介导的Bcl-2 Ser70位点的磷酸化,在肝癌细胞的存活和抗药中发挥重要作用。抑制Bcl-2能够促进化疗药物5-FU介导的HCC细胞凋亡,该结果为治疗GCDA介导的耐药性肝癌提供新的思路。     

3-(2-Methoxytetrahydrofuran-2-yl)pyrazoles: a novel class of potent, selective cyclooxygenase-2 (COX-2) inhibitors

A series of 3-(2-methoxytetrahydrofuran-2-yl)pyrazoles (4–10) was synthesized. The compounds were evaluated for their ability to inhibit cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) activity in human whole blood (HWB). The compound, 5-(4-methanesulfonylphenyl)-3-(2-methoxytetrahydrofuran-2-yl)-1-p-tolyl-1H-pyrazole 5 showed potent and selective COX-2 inhibition (IC₅₀ for COX-1: >100 μM and COX-2: 1.2 μM).     

人白细胞介素－2的两个部分拮抗剂

通过定点诱变技术得到6个生物活性剧烈下降的人白细胞介素-2(IL-2)突变体,其中两个突变体即15Val-IL-2和126Asp-IL-2可以在一定浓度范围内使IL-2的生物效应降低.在对高亲和力IL-2受体(IL-2R)的竞争抑制实验中,15Val-IL-2和126Asp-IL-2又表现了一定的竞争能力.这些结果表明15Val-IL-2和126Asp-IL-2的部分拮抗天然IL-2的作用.结合IL-2二级结构分析及对IL-2与IL-2R相互作用的已有认识,可认为15Val-IL-2和126Asp-IL-2的部分拮抗作用产生的原因在于替换残基在空间上对IL-2与IL-2R βγ亚基结合微环境的轻微扰动,干扰了IL-2有关残基与IL-2R βγ亚基的结合,但尚不能完全阻止其与IL-2R βγ亚基的结合.     

IL－2部分拮抗剂可用于IL－2突变体对受体亚基结合能力的初步鉴定

用定点突变法分别得到了两个人白细胞介素-2(IL-2)的部分拮抗剂15Val-IL-2和126Asp-IL-2以及一个为IL-2受体α亚基结合缺陷型的突变体62Leu-IL-2,当将15Val-IL-2或126Asp-IL-2与62Leu-IL-2共同保温时,62Leu-IL-2的活性受到明显抑制,对此现象机理的分析表明15Val-IL-2或126Asp-IL-2可用于IL-2受体亚基结合缺陷型突变体的初步鉴定.同时,这一思路在其它受体-配基系统中具有一定的适用性.     

大肠杆菌表达的重组白细胞介素-2的初步纯化

本文对大肠杆菌表达产生的重组白细胞介素-2进行了纯化研究。通过比较两种方法制备的rIL-2包含体的纯度,发现用4mol/L脲溶解可溶性细菌蛋白后可使rIL-2包含体纯度达70％;在高浓度变性剂条件下进行凝胶过滤,解决了rIL-2易聚合的问题;结合透析,利用空气氧化形成高活性氧化型rIL-2;经SephadexG-100凝胶过滤,DEAE离子交换等步骤纯化,得到了均一性rIL-2,纯度达98％,比活达4.3×10~6u/mg蛋白,得率为30.8％。     

大肠癌组织中TS和COX-2的表达及其与患者无病生存期的关系

目的:探讨大肠癌患者癌组织中环氧化酶-2(COX-2)、胸苷酸合成酶(TS)的表达及其与患者无病生存期的关系。方法:筛选我院收治的大肠癌根治术患者,选择无病生存期大于48个月者30例和无病生存期小于48个月者29例。采用免疫组化法检测大肠癌组织中COX-2和TS的表达,并分析其与患者无病生存期的关系。结果:49例结直肠癌患者中,TS的阳性表达率为91.84%,COX-2的阳性表达率为77.55%。不同无病生存期的大肠癌患者TS的表达水平比较无统计学差异(P=0.646)。COX-2在无病生存期48个月的患者癌组织中表达水平明显低于无病生存期48个月的患者,差异有统计学意义(P=0.033)。结论:COX-2与大肠癌患者的无病生存期显著相关,可能成为预测大肠癌预后的参考指标。     

Bioaugmentation of a rotating biological contactor for degradation of 2-fluorophenol

The performance of a laboratory scale rotating biological contactor (RBC) towards shock loadings of 2-fluorophenol (2-FP) was investigated. During a period of ca. 2 months organic shock loadings of 25 mg L?1 of 2-FP were applied to the RBC. As no biodegradation of 2-FP was observed, bioaugmentation of the RBC with a 2-FP degrading strain was carried out and, along ca. 6 months, organic shock loadings within a range of 25-200 mg L?1 of 2-FP were applied. Complete biodegradation of 50 mg L?1 of 2-FP was observed during operation of the reactor. The RBC showed to be robust towards starvation periods, as after ca. 1month of non-supply of the target compound, the reactor resumed 2-FP degradation. The inoculated strain was retained within the biofilm in the disks, as the 2-FP degrading strain was recovered from the biofilm by the end of the experiment, thus bioaugmentation was successfully achieved.     

Therapy with interleukin-2 induces the systemic release of phospholipase-A2

Therapy with interleukin-2 (IL-2) induces remissions in some forms of cancer. This treatment however, is accompanied by side-effects which, in part, may be mediated by the formation of eicosanoids and plateletactivating factor. We investigated the systemic release of phospholipase A₂ (PLA₂), a rate-limiting enzyme in the formation of these lipid mediators, in patients receiving IL-2. In a pilot study of 4 patients we observed an increase in PLA₂ activity in serial plasma samples obtained during the first day after a bolus infusion of IL-2, which increase closely correlated with that of antigen levels of secretory phospholipase A₂ (sPLA₂) as measured by enzyme-linked immunosorbent assay (r=0.92;P<0.001). In 20 patients, receiving 12×10⁶–18×10⁶ IU IL-2/m², we then investigated the course of antigenic levels of sPLA₂ in relation to those of the cytokines tumour necrosis factor (TNF) and interleukin-6 (IL-6) (both cytokines may induce sPLA₂ in vivo). From 4 h on, sPLA₂ levels significantly increased, reaching a peak 24 h after the IL-2 infusion. Subsequent IL-2 infusions even induced a further increase of sPLA₂. This increase of sPLA₂ was presumably not due to a direct effect of IL-2 on, for example, hepatocytes, since this cytokine, in contrast to IL-1, IL-6, TNF and interferon , was not able to induce the synthesis of sPLA₂ by Hep G2 cells in vitro. Consistent with this, plasma levels of TNF and IL-6 in the patients rose, reaching peak levels before a zenith of sPLA₂ occurred, i.e at 2 h and 4 h after the start of the IL-2 infusion respectively. sPLA₂ levels significantly correlated with the development of the side-effects increase in body weight (r=0.49;P<0.0001) and decrease in mean arterial blood pressure (r=0.40;P<0.0001). Moreover, maximum sPLA₂ levels induced by IL-2 were higher in patients who had progressive disease after therapy than in patients who had stable disease or a partial response.     

Differential expression of cyclin G2, cyclin-dependent kinase inhibitor 2C and peripheral myelin protein 22 genes during adipogenesis

Increase of fat cells (FCs) in adipose tissue is attributed to proliferation of preadipocytes or immature adipocytes in the early stage, as well as adipogenic differentiation in the later stage of adipose development. Although both events are involved in the FC increase, they are contrary to each other, because the former requires cell cycle activity, whereas the latter requires cell cycle withdrawal. Therefore, appropriate regulation of cell cycle inhibition is critical to adipogenesis. In order to explore the important cell cycle inhibitors and study their expression in adipogenesis, we adopted a strategy combining the Gene Expression Omnibus (GEO) database available on the NCBI website and the results of quantitative real-time PCR (qPCR) data in porcine adipose tissue. Three cell cycle inhibitors – cyclin G2 (CCNG2), cyclin-dependent kinase inhibitor 2C (CDKN2C) and peripheral myelin protein (PMP22) – were selected for study because they are relatively highly expressed in adipose tissue compared with muscle, heart, lung, liver and kidney in humans and mice based on two GEO DataSets (GDS596 and GDS3142). In the latter analysis, they were found to be more highly expressed in differentiating/ed preadipocytes than in undifferentiated preadipocytes in human and mice as shown respectively by GDS2366 and GDS2743. In addition, GDS2659 also suggested increasing expression of the three cell cycle inhibitors during differentiation of 3T3-L1 cells. Further study with qPCR in Landrace pigs did not confirm the high expression of these genes in adipose tissue compared with other tissues in market-age pigs, but confirmed higher expression of these genes in FCs than in the stromal vascular fraction, as well as increasing expression of these genes during in vitro adipogenic differentiation and in vivo development of adipose tissue. Moreover, the relatively high expression of CCNG2 in adipose tissue of market-age pigs and increasing expression during development of adipose tissue was also confirmed at the protein level by western blot analysis. Based on the analysis of the GEO DataSets and results of qPCR and Western blotting we conclude that all three cell cycle inhibitors may inhibit adipocyte proliferation, but promote adipocyte differentiation and hold a differentiated state by inducing and maintaining cell cycle inhibition. Therefore, their expression in adipose tissue is positively correlated with age and mature FC number. By regulating the expression of these genes, we may be able to control FC number, and, thus, reduce excessive fat tissue in animals and humans.     

PS-2基因的克隆及其在肝癌中的表达

利用荧光差异显示技术比较了正常肝、肝硬化和肝癌组织 m RNA的表达 ,1 4个有差异的条带通过 Northern blot分析表明其中 9个为阳性 .令人感兴趣的是一个～ 5 0 0 bp的 c DNA片段 ,它在正常肝和肝硬化中低表达 ,在肝癌组织中高表达 .通过测序 ,发现该片段与 PS- 2 ( presenilin- 2 )基因有 94 %的同源性 .PS- 2基因的突变与早发性阿尔茨海默氏症有关 ,但在肝癌发生中的作用未明 .也许 PS- 2基因的上调涉及到肝癌发生的分子机理     

前列腺素D2及其受体与哺乳动物生殖

前列腺素D2(PGD2)是前列腺素(PGs)家族成员之一,广泛分布于各种哺乳动物组织中,并发挥多种生理功能。PGD,可以促进睡眠、诱导过敏反应、抑制血小板凝集及松弛平滑肌等,并且在生殖系统中起重要作用。机体中存在生化和免疫功能截然不同的两类前列腺素D合成酶(PGDs)：脑型PGDS(L-PGDS)和生血型PGDS(hPGDS)。生殖系统中,L-PGDS主要存在于雄性生殖道,可能在睾丸发育、精子发生、精子成熟以及血一睾和血一附睾屏障等方面发挥重要作用。hPGDS在妊娠时期的子宫内膜和胚胎滋养层中表达,由其产生的PGD2可能通过DP和CRTH2两种受体来维持妊娠。此外,PGD2还可能与女性不孕以及精子在雌性生殖道内的运输等有关。     

Host variations in SARS-CoV-2 infection

The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the zoonotic pathogen that causes the “Coronavirus Disease of 2019 (COVID-19)”, and COVID-19 itself is yet to be thoroughly understood. Both the disease as well as the mechanisms by which the host interacts with the SARS-CoV-2 have not been fully enlightened. The epidemiological factors –e.g. age, sex, race-, the polymorphisms of the host proteins, the blood types and individual differences have all been in discussions about affecting the progression and the course of COVID-19 both individually and collectively, as their effects are mostly interwoven. We focused mainly on the effect of polymorphic variants of the host proteins that have been shown to take part in and/or affect the pathogenesis of COVID-19. Additionally, how the procedures of diagnosing and treating COVID-19 are affected by these variants and what possible changes can be implemented are the other questions, which are sought to be answered.     

环氧化物水解酶2在肝细胞癌中的表达水平与功能作用

本研究通过公共数据和实验数据,全面分析环氧化物水解酶2(epoxide hydrolase 2, EPHX2)在肝细胞癌中的表达情况、功能作用以及预后意义。利用GEO和MitoCarta数据集,筛选肝细胞癌中呈差异表达的线粒体相关基因;利用TCGA数据库分析EPHX2及其相关基因在肝细胞癌中的表达水平;运行R包绘制Kaplan-Meier生存曲线和功能富集分析;基于STRING和GSEA构建蛋白质互作网络和基因集富集分析;荧光定量PCR和GEO数据集验证EPHX2在肝细胞癌中的表达水平。本研究共筛选得到15个在肝细胞癌中呈差异表达的线粒体相关基因。EPHX2在肝细胞癌组织中的表达水平显著降低(P<0.01)。EPHX2表达水平与肝癌患者性别、分期和级别有关,而与年龄、T分期等因素无关。与EPHX2低表达组肝癌患者相比,EPHX2高表达组肝癌患者预后较好。功能富集结果显示,EPHX2与补体途径、脂肪酸降解等信号通路有关。蛋白质互作网络结果显示,EPHX2与HAO1、AGXT、ACOX1、GSTκ1、SCP-2、CAT、CYP2C8,CYP2C9,CYP2B6,和CYP2J2等密切相关。GSEA结果显示,EPHX2低表达组与肝癌细胞增殖、肝癌复发等基因集正相关。荧光定量PCR和GEO数据集验证结果显示,EPHX2在肝细胞癌组织和肝癌细胞株中呈显著低表达。EPHX2在肝细胞癌中呈显著低表达,提示其可能在肝细胞癌发生发展过程中发挥抑癌基因作用,但具体作用机制还需进一步验证。     

LINC00152 promotes the proliferation of gastric cancer cells by regulating B-cell lymphoma-2

LINC00152 has been considered to be associated with the tumorigenesis and the occurrence of gastric cancer; however, the mechanism of LINC00152 has yet to be fully elucidated. In the present study, the expression levels of LINC00152 in tissues, serum, and peripheral blood mononuclear cells (PBMCs) of patients with gastric cancer were determined using real-time polymerase chain reaction. The functions of LINC00152 with respect to the proliferation, apoptosis, migration, and invasive abilities of the gastric cancer cells were evaluated by cell proliferation analysis, flow cytometry, cell scratch wound assay, and transwell migration experiments. A mouse xenotransplant model of gastric tumors was established to detect the role of LINC00152 in vivo, and the expression levels of B-cell lymphoma-2 (Bcl-2) family proteins were investigated by Western blot analysis. The results revealed that LINC00152 was overexpressed in tissues, serum, and PBMCs of patients with gastric cancer. Moreover, LINC00152 could promote the migration and invasive abilities and suppress the apoptosis, of gastric cancer cells through regulating the Bcl-2 protein family. LINC00152 could bind with Bcl-2 directly to induce the activation of cell cycle signaling, and this may be a potential target for the therapy of gastric cancer in the future.     

超临界CO2技术萃取蛋黄磷脂

采用新型物理分离技术──超临界CO₂萃取法,提取天然蛋黄粉中的磷脂.在40MPa,先去除蛋黄粉中甘油三酯和胆固醇,再萃取磷脂.结果显示,磷脂纯度为95%,N/P比值为1.003,λ_max=214nm,薄层层析显示磷脂着色点清晰,并去除了绝大部分甘油三酯和胆固醇.此法操作简单、产品质量高、安全和不污染环境,还可得到天然纯蛋黄油和蛋白.