Structures of Sarcorucinine D and Pachyaximine A,B

In this paper, three new alkaloids, sarcorucinine D (1), and paehyaximine A, B (2, 3) were isolated from Sarcococca ruscifolia Stapf and Pachysandra axillaris Franch, respectively. The structures of sarcorucinine D (1) and pachyaximine A, B (2, 3) were elucidated to be 3β-hydroxyl-20α-dimethylamino-5α-pregnane (1); and 3β-methoxyl-20α-dimethylamino-pregne-5-ne (2); and 3β-methoxyl-16-hydroxyl-20α-dimethylaminopregne-5-ne (16-hydroxyl-pachyaxi- mine A) (3), respectively.     

低氧对巨噬细胞分泌TNF-α和IL-6的影响及其机制

目的:观察低氧对巨噬细胞(Mφ)前炎症因子TNF-α和IL-6分泌的影响及其机制.方法:收集分离小鼠腹腔Mφ,建立Mφ的低氧(1% O2,5%CO2)培养模型,并用非特异性酯酶染色法进行鉴定;ELISA法检测上清液中TNF-α和IL-6的含量;RT-PCR法检测TNF-α和IL-6的转录物水平;用Western blot法检测Mφ核内NF-κB的激活量;通过在培养液中加入氢化可的松(5 mg/L),观察低氧时TNF-α和IL-6分泌量的变化.结果:TNF-α和IL-6分泌量在低氧12 h时明显增加(P＜0.01);低氧6 h时,TNF-α mRNA和IL-6 mRNA表达量明显高于对照组(P＜0.01);M中核内NF-κB的激活量在低氧2 h时明显增高(P＜0.05),低氧5 h内持续存在;而当培养液中加入氢化可的松抑制NF-κB活性后,TNF-α和IL-6的分泌水平无明显变化.结论:低氧可通过核转录因子NF-κB途径促进细胞因子TNF-α和IL-6基因的表达和分泌.     

Differential effects of interleukin-7 and interleukin-15 on NK cell anti-human immunodeficiency virus activity

The ability of interleukin-7 (IL-7) and IL-15 to expand and/or augment effector cell functions may be of therapeutic benefit to human immunodeficiency virus (HIV)-infected patients. The functional effects of these cytokines on innate HIV-specific immunity and their impact on cells harboring HIV are unknown. We demonstrate that both IL-7 and IL-15 augment natural killer (NK) function by using cells (CD3(-) CD16(+) CD56(+)) from both HIV-positive and -negative donors. Whereas IL-7 enhances NK function through upregulation of Fas ligand, the effect of IL-15 is mediated through upregulation of tumor necrosis factor-related apoptosis-inducing ligand. The difference in these effector mechanisms is reflected by the ability of IL-15-treated but not IL-7-treated NK cells to reduce the burden of replication-competent HIV in autologous peripheral blood mononuclear cells (PBMC) (infectious units per million for control NK cells, 6.79; for IL-7-treated NK cells, 236.17; for IL-15-treated cells, 1.01; P = 0.01 versus control). In addition, the treatment of PBMC with IL-15-treated but not IL-7-treated NK cells causes undetectable HIV p24 (five of five cases), HIV RNA (five of five cases), or HIV DNA (three of five cases). These results support the concept of adjuvant immunotherapy of HIV infection with either IL-7 or IL-15 but suggest that the NK-mediated antiviral effect of IL-15 may be superior.     

Association between Obesity and Cardiometabolic Health Risk in Asian-Canadian Sub-Groups

Objectives

To quantify and compare the association between the World Health Organizations’ Asian-specific trigger points for public health action [‘increased risk’: body mass index (BMI) ≥23 kg/m², and; ‘high risk’: BMI ≥27.5 kg/m²] with self-reported cardiovascular-related conditions in Asian-Canadian sub-groups.

Methods

Six cycles of the Canadian Community Health Survey (2001–2009) were pooled to examine BMI and health in Asian sub-groups (South Asians, Chinese, Filipino, Southeast Asians, Arabs, West Asians, Japanese and Korean; N = 18 794 participants, ages 18–64 y). Multivariable logistic regression, adjusting for demographic, lifestyle characteristics and acculturation measures, was used to estimate the odds of cardiovascular-related health (high blood pressure, heart disease, diabetes, ‘at least one cardiometabolic condition’) outcomes across all eight Asian sub-groups.

Results

Compared to South Asians (OR = 1.00), Filipinos had higher odds of having ‘at least one cardiometabolic condition’ (OR = 1.29, 95% CI: 1.04–1.62), whereas Chinese (0.63, 0.474–0.9) and Arab-Canadians had lower odds (0.38, 0.28–0.51). In ethnic-specific analyses (with ‘acceptable’ risk weight as the referent), ‘increased’ and ‘high’ risk weight categories were the most highly associated with ‘at least one cardiometabolic condition’ in Chinese (‘increased’: 3.6, 2.34–5.63; ‘high’: 8.9, 3.6–22.01). Compared to normal weight South Asians, being in the ‘high’ risk weight category in all but the Southeast Asian, Arab, and Japanese ethnic groups was associated with approximately 3-times the likelihood of having ‘at least one cardiometabolic condition’.

Conclusion

Differences in the association between obesity and cardiometabolic health risks were seen among Asian sub-groups in Canada. The use of WHO’s lowered Asian-specific BMI cut-offs identified obesity-related risks in South Asian, Filipino and Chinese sub-groups that would have been masked by traditional BMI categories. These findings have implications for public health messaging, especially for ethnic groups at higher odds of obesity-related health risks.     

Se和V_E与克山病

以克山病病区粮配成基础饲料,另在基础饲料中分别补充Ｓｅ或ＶＥ,或Ｓｅ＋ＶＥ喂养大鼠,在细胞及亚细胞水平上以Ｃａ代谢为主研究并比较了Ｓｅ和ＶＥ在克山病病因中的作用。测量了心肌细胞和心肌线粒体的Ｃａ代谢及有关指标、心肌线粒体能量转换功能及心肌组织自由基含量。结果表明,在低Ｓｅ病区粮中补充Ｓｅ或ＶＥ均能在一定程度上预防病区粮中致病因素对心肌细胞及线粒体的损伤;并且补充Ｓｅ或ＶＥ均能使心肌组织中自由基含量减少。提示Ｓｅ和ＶＥ是通过清除体内过量自由基预防细胞和线粒体的损伤的。但值得注意的是,实验中所用病区粮ＶＥ含量不低于甚至高于非病区对照粮,在低Ｓｅ情况下,所补ＶＥ的量需要相当大（如本实验中补充２００μｇ／ｇ）才能较明显地预防心肌细胞和心肌线粒体的损伤。通过对这些结果的分析,进一步肯定低Ｓｅ是克山病形成的重要因素之一。     

镰刀菌Q7-31T内切葡聚糖酶Egn21的分离纯化及酶学性质

【目的】为进一步研究镰刀菌Q7-31T产生的植物细胞壁降解酶的酶系信息。【方法】以1%(W/V)蛋白胨为氮源,0.5%(W/V)燕麦秸秆为碳源,20°C、120 r/min振荡培养3 d,诱导发酵培养菌株,获得的粗酶液经过Sephacry S-100凝胶柱层析和DEAE琼脂糖弱阴离子交换柱层析,最终得到纯化的内切葡聚糖酶,并对其进行酶学性质分析及串联质谱鉴定。【结果】研究表明:Egn21的分子质量为44.25 kDa,等电点为4.91;酶学特性研究显示:Egn21降解羧甲基纤维素的最适反应温度为40°C,在45°C以下比较稳定。该酶最适pH为6.0,在pH为5.0–8.0条件之间比较稳定。Co~(2+)、Zn~(2+)和Mg~(2+)对其没有明显作用,而Fe~(2+)、Ca~(2+)、K~(+)、Na~(+)和Mn~(2+)对酶活性有抑制作用,Hg~(2+)会使酶失去活性。【结论】从Q7-31T中分离纯化得到的内切葡聚糖酶Egn21,经过酶学特性与串联质谱鉴定结果显示其属于GH5家族。     

Small molecules blocking the entry of severe acute respiratory syndrome coronavirus into host cells

Severe acute respiratory syndrome coronavirus (SARS-CoV) is the pathogen of SARS, which caused a global panic in 2003. We describe here the screening of Chinese herbal medicine-based, novel small molecules that bind avidly with the surface spike protein of SARS-CoV and thus can interfere with the entry of the virus to its host cells. We achieved this by using a two-step screening method consisting of frontal affinity chromatography-mass spectrometry coupled with a viral infection assay based on a human immunodeficiency virus (HIV)-luc/SARS pseudotyped virus. Two small molecules, tetra-O-galloyl-beta-D-glucose (TGG) and luteolin, were identified, whose anti-SARS-CoV activities were confirmed by using a wild-type SARS-CoV infection system. TGG exhibits prominent anti-SARS-CoV activity with a 50% effective concentration of 4.5 microM and a selective index of 240.0. The two-step screening method described here yielded several small molecules that can be used for developing new classes of anti-SARS-CoV drugs and is potentially useful for the high-throughput screening of drugs inhibiting the entry of HIV, hepatitis C virus, and other insidious viruses into their host cells.     

Host S-nitrosylation inhibits clostridial small molecule-activated glucosylating toxins

The global prevalence of severe Clostridium difficile infection highlights the profound clinical significance of clostridial glucosylating toxins. Virulence is dependent on the autoactivation of a toxin cysteine protease, which is promoted by the allosteric cofactor inositol hexakisphosphate (InsP(6)). Host mechanisms that protect against such exotoxins are poorly understood. It is increasingly appreciated that the pleiotropic functions attributed to nitric oxide (NO), including host immunity, are in large part mediated by S-nitrosylation of proteins. Here we show that C. difficile toxins are S-nitrosylated by the infected host and that S-nitrosylation attenuates virulence by inhibiting toxin self-cleavage and cell entry. Notably, InsP(6)- and inositol pyrophosphate (InsP(7))-induced conformational changes in the toxin enabled host S-nitrosothiols to transnitrosylate the toxin catalytic cysteine, which forms part of a structurally conserved nitrosylation motif. Moreover, treatment with exogenous InsP(6) enhanced the therapeutic actions of oral S-nitrosothiols in mouse models of C. difficile infection. Allostery in bacterial proteins has thus been successfully exploited in the evolutionary development of nitrosothiol-based innate immunity and may provide an avenue to new therapeutic approaches.     

大鼠三叉神经节不同直径神经元ATP-激活电流的特征

目的:探讨大鼠三叉神经节不同直径神经元ATP-激活电流的特征。方法:应用全细胞膜片钳技术进行实验。结果:①92.3%(60/65)的细胞对ATP敏感,有反应的细胞可记录到三种型式的ATP-激活电流:快速激活快速失活型(Fast type,F型)、快速激活缓慢失活型(Intermediate type,I型)和缓慢激活缓慢失活型(Slowtype,S型)。三种电流均具有浓度依赖性。②小直径的细胞多表现为F型特征,大直径的细胞多表现为S型特征,而中等大小的细胞多表现为I型特征。③动力学特征:三种类型的ATP激活电流上升相从10%到90%的时间:F型:(33.6±4.5)ms;Ⅰ型:(62.2±9.9)ms;S型:(302.1±62)ms。去敏感相从10%到90%的时间:F型:(399.4±58.2)ms;S型:>500ms。④I-V曲线:三种电流均表现为内向整流的特性,而且翻转电位均为0~5mV。⑤量-效关系:Ⅰ型的量-效曲线居中间,F型的下移,S型的上移,三种类型电流量-效曲线的EC50非常接近。结论:三种型式的ATP-激活电流可能是由不同亚单位组合的P2X受体各亚型所介导,这些亚型分布于不同大小的三叉神经节神经元,从而传导不同的信息。     

Characterization of 9-nitrocamptothecin liposomes: anticancer properties and mechanisms on hepatocellular carcinoma in vitro and in vivo

Background

Hepatocellular carcinoma (HCC) is the third most common cause of cancer related mortality worldwide. 9-Nitrocamptothecin (9NC) is a potent topoisomerase-I inhibitor with strong anticancer effect. To increase the solubility and stability, we synthesized a novel 9NC loaded liposomes (9NC-LP) via incorporating 9NC into liposomes. In the present study, we determined the effects of 9NC and 9NC-LP on in vitro and in vivo, and the underlying mechanisms.

Methodology/Principal Findings

We first analyzed the characteristics of 9NC-LP. Then we compared the effects of 9NC and 9NC-LP on the proliferation and apoptosis of HepG2, Bel-7402, Hep3B and L02 cells in vitro. We also investigated their anticancer properties in nude mice bearing HCC xenograft in vivo. 9NC-LP has a uniform size (around 190 nm) and zeta potential (∼−11 mV), and exhibited a steady sustained-release pattern profile in vitro. Both 9NC and 9NC-LP could cause cell cycle arrest and apoptosis in a dose-dependent and p53-dependent manner. However, this effect was not ubiquitous in all cell lines. Exposure to 9NC-LP led to increased expression of p53, p21, p27, Bax, caspase-3, caspase-8, caspase-9 and apoptosis-inducing factor, mitochondrion-associated 1 and decreased expression of Bcl-2, cyclin E, cyclin A, Cdk2 and cyclin D1. Furthermore, 9NC-LP exhibited a more potent antiproliferative effect and less side effects in vivo. Western blot analysis of the xenograft tumors in nude mice showed similar changes in protein expression in vivo.

Conclusions/Significance

In conclusion, 9NC and 9NC-LP can inhibit HCC growth via cell cycle arrest and induction of apoptosis. 9NC-LP has a more potent anti-tumor effect and fewer side effects in vivo, which means it is a promising reagent for cancer therapy via intravenous administration.