IKK�� downregulation is critical for triggering JNKs-dependent cell apoptotic response in the human hepatoma cells under arsenite exposure

Arsenite has a long history in treating leukemia, which might be also effective in the therapy of other cancers. Our previous published data have demonstrated that arsenite exposure induces apoptosis in the HepG2 human hepatoma cells via activating JNKs/AP-1 pathway, but the upstream signaling events responsible for JNKs (c-Jun N-terminal kinase) cascade activation have not been fully discovered. Since cross-talk between IKK/NF-κB and JNKs pathways under stress conditions is a hot topic, in this article, we investigate the potential roles of IKKα and IKKβ, the catalytic subunits of IKK complexes, in the arsenite-induced JNKs pathway activation in the HepG2 cells. We found that arsenite exposure induced JNKs and AP-1 activation accompanying with a significant reduction of both IKKα and IKKβ expressions. Overexpression of IKKβ, but not of IKKα, inhibited arsenite-induced MKK7/JNKs/AP-1 pathway activation as well as the apoptotic response. Therefore, we conclude that the downregulation of IKKβ expression is the prerequisite signaling event for mediating JNKs pathway activation and the cellular apoptotic response in the HepG2 cells under arsenite exposure. Targeting IKKβ might be helpful to enhance the tumor therapeutic effect of arsenite.     

高、低氮浓度对2株真眼点藻的生长和油脂积累的影响

【目的】研究氮浓度对真眼点藻纲(Eustigmatophyceae)的2株高产油微藻大真眼点藻(Eustigmatos magnus,EM)和波氏真眼点藻(Eustigmatos polyphem,EP)的细胞形态、生长、总脂含量、脂质组成和脂肪酸组成与含量的时序变化规律。【方法】利用高氮(18.0 mmol/L NO3?-N)和低氮(3.6 mmol/L NO3?-N)浓度培养微藻。【结果】形态观察结果表明,大真眼点藻(E. magnus)和波氏真眼点藻(E. polyphem)营养细胞具有1个周生的裂叶状叶绿体,细胞质中有液泡,内含能够振动的颗粒物,以及一个较为明显的红色色素体;生殖方式通过形成2个D形或4个四角形的似亲孢子;随着培养周期的延伸和营养盐的消耗,细胞中油体逐步形成,其数量不断增加,体积不断增大。实验结果表明,初始氮浓度对2种微藻的总脂积累及生长均有显著影响(P<0.05),低氮浓度下2种微藻的生物质浓度分别为9.0 g/L和8.5 g/L,均低于高氮浓度下的生物质浓度。而低氮浓度下2种微藻的总脂、中性脂和总脂肪酸的含量以及总脂、中性脂与总脂肪酸的单位体积产率均明显高于高氮浓度组,其最高值分别为：59.10%、51.90%、46.95%和0.28、0.24、0.22 g/(L·d) (EM);64.20%、56.80%、50.01%和0.32、0.28、0.25 g/(L·d) (EP)。脂肪酸分析结果表明,两种微藻的脂肪酸主要成分均为棕榈酸(C16:0)、棕榈油酸(C16:1)、油酸(C18:1)和二十碳五烯酸(C20:5,EPA),四者的总含量(占总脂肪酸)分别达到85.83%和85.48%,其中棕榈油酸的含量最高。【结论】低氮浓度胁迫有利于大真眼点藻和波氏真眼点藻细胞内油脂的积累,两种微藻均为适合于生产生物柴油的油脂生产藻株。     

Protein preparation, crystallization and preliminary X-ray crystallographic analysis of N-acetylglutamate kinase from Streptococcus mutans

The N-acetylglutamate kinase from Streptococcus mutans was expressed in Escherichia coli in soluble form and purified to homogeneity. Crystals suitable for X-ray diffraction were obtained by hanging-drop vapor diffusion method and diffracted to 2.06 A. The crystal belonged to space group P2(1)2(1)2, with unit cell parameters a = 57.19 A, b =94.76 A, c =47.58 A. The gel filtration and initial phasing results showed that the enzyme exists as a monomer, which is different from previously reported N-acetylglutamate kinases.     

Alterations in methylation and expression levels of imprinted genes H19 and Igf2 in the fetuses of diabetic mice

The study aimed to reveal alterations in expression and methylation levels of the growth-related imprinted genes H19 and Igf2 in fetuses of diabetic mice. Diabetes was induced in female mice by intraperitoneal injection of streptozotocin. DNA and total RNA were extracted from fetuses obtained from diabetic and control dams on embryonic day (E) 14. Real-time RT-PCR analysis revealed that the mRNA expression of Igf2 in fetuses from diabetic mice was 0.65-fold of the control counterparts. Bisulfite genomic sequencing demonstrated that the methylation level of the H19-Igf2 imprint control region was 19.1% higher in diabetic fetuses than in those of control dams. In addition, the body weight of pups born to diabetic dams was 26.5% lower than that of the control group. The results indicate that maternal diabetes can affect fetal development by means of altered expression of imprinted genes. The modified genomic DNA methylation status of imprinting genes may account for the change in gene expression.     

Microarray analysis of gene expression profile in resistant and susceptible Bombyx mori strains reveals resistance-related genes to nucleopolyhedrovirus

To investigate the molecular mechanism of silkworm resistance to BmNPV infection, we constructed a near-isogenic line (BC8) with BmNPV resistance using highly resistant (NB) and highly susceptible parental strains (306). We investigated variations in the gene expression in the midguts of BmNPV-infected BC8 and 306 at 12 h pi using the microarray. 92 differentially expressed genes were identified. Real-time qPCR analysis confirmed that 10 genes were significantly up-regulated or down-regulated in the midguts of BC8 and NB compared to 306. To our knowledge, we first defined the role of the amino acid transporter and 26S proteasome in insect antiviral. However, serine protease was not completely consistent with data of reported previously in insect antiviral. The role of the 5 genes (Bm123, Bm122, COP β′, aquaporin, glycoside hydrolases) was also demonstrated in insect antiviral. Our results provided new insights into the molecular mechanism of the Bombyx mori immune response against BmNPV infection.     

康莱特联合阿霉素诱导骨肉瘤细胞凋亡的实验研究

目的研究康莱特对骨肉瘤细胞的诱导凋亡作用及其相关机制,探寻骨肉瘤的中西医结合化疗方案.方法免疫组化及RT-PCR方法检测不同浓度康莱特作用下骨肉瘤细胞的Fas表达,并将康莱特与阿霉素单独及联合应用于骨肉瘤细胞,采用MTT法检测细胞毒性作用,流式细胞仪定量分析凋亡细胞所占比例,倒置相差显微镜、荧光显微镜下观察凋亡细胞形态改变.结果康莱特可上调骨肉瘤细胞的Fas表达并诱导凋亡,康莱特浓度达到10μl/ml时,细胞抑制率不再明显改变,但如与1μlg/ml阿霉素合用将使细胞抑制率进一步提升(P<0.01).细胞形态观察及凋亡率测定也显示,康莱特与阿霉素联用可诱导更多骨肉瘤细胞凋亡.结论康莱特可通过上调Fas表达诱导骨肉瘤细胞凋亡,康莱特与阿霉素合用可提高杀伤骨肉瘤细胞.     

Knowledge of HIV and Willingness to Conduct Oral Rapid HIV Testing among Dentists in Xi’an China

Introduction

China is considered a country of low HIV prevalence (780,000 people living with HIV), however, HIV infections among high-risk populations continue to grow at alarming rates. Voluntary Counseling and Testing services were first implemented in 2003, and oral rapid HIV testing (ORHT) began in 2012. Dentists, as oral health experts, would be well placed to conduct ORHT. We assessed willingness of dentists to undertake ORHT in their clinical practice.

Methods

A cross-sectional, paper-based survey of dentists from the Xi’an region of China was conducted from April to June 2013. Dentists were recruited from Shaanxi Stomatological Association using a stratified sampling methodology. A 40-item survey was used to measure knowledge of HIV, attitudes toward people living with HIV and willingness to conduct ORHT.

Results

477 dentists completed the survey with a mean HIV knowledge test score of 13.2/18 (SD 1.9). If made available in the dental setting, 276 (57.9%) preferred to use blood to diagnose HIV, only 190 (39.8%) preferred saliva or both. Four hundred and thirty-five (91.2%) thought that ORHT was needed in dental clinics. Female dentists felt more accepting of ORHT than males (93.8% vs. 87.8%; χ²=5.145; p<0.05). 42.6% of the participants who responded thought that lack of education on ORHT for dentists was the most urgent problem to solve for ORHT, 144 (31.3%) thought that lack of support for ORHT from patients was the most urgent problem. There was statistically significant difference among dental hospital, dentistry and department of dentistry (χ²=24.176; p<0.05).

Conclusions

The majority of Chinese dentists thought that ORHT was needed in the dental setting. Providing opportunities for dentists and dental students to learn about HIV testing guidelines and practices is needed as well as feasibility and implementation science research.     

Aldolase A Enhances Intrahepatic Cholangiocarcinoma Proliferation and Invasion through Promoting Glycolysis

Energy metabolism reprogramming has been implicated in tumorigenesis and development. Key metabolism enzyme Aldolase A (ALDOA) has been shown to be highly expressed and involved in various kinds of cancers including hepatocellular carcinoma. In this study, we found that ALDOA was highly expressed in clinical intrahepatic cholangiocarcinoma (ICC) tissues, and its high expression was negatively correlated with overall survival (OS) and recurrence-free survival (RFS) in ICC patients. Knockdown of ALDOA expression significantly inhibited the proliferation and migration of ICC both in vitro and in vivo, while highly-expressed ALDOA in ICC cells promoted the proliferation and migration of ICC cells. By applying ALDOA inhibitor and metabolic mass spectrometry tests, we demonstrated that ALDOA modulated the biological characteristics and metabolic level of ICC cells depending on its enzymatic activity. In summary, ALDOA promotes ICC proliferation and migration by enhancing ICC cells glycolysis. Blocking enzymatic activity of ALDOA provides a strategy to inhibit ICC.     

Laccase-catalyzed oxidation of phenolic compounds in organic media

Rhus vernificera laccase-catalyzed oxidation of phenolic compounds, i.e., (+)-catechin, (−)-epicatechin and catechol, was carried out in selected organic solvents to search for the favorable reaction medium. The investigation on reaction parameters showed that optimal laccase activity was obtained in hexane at 30 °C, pH 7.75 for the oxidation of (+)-catechin as well as for (−)-epicatechin, and in toluene at 35 °C, pH 7.25 for the oxidation of catechol. E_a and Q₁₀ values of the biocatalysis in the reaction media of the larger log p solvents like isooctane and hexane were relatively higher than those in the reaction media of lower log p solvents like toluene and dichloromethane. Maximum laccase activity in the organic media was found with 6.5% of buffer as co-solvent. A wider range of 0–28 μg protein/ml in hexane than that of 0–16.7 μg protein/ml in aqueous medium was observed for the linear increasing conversion of (+)-catechin. The kinetic studies revealed that in the presence of isooctane, hexane, toluene and dichloromethane, the K_m values were 0.77, 0.97, 0.53 and 2.9 mmol/L for the substrate of (+)-catechin; 0.43, 0.34, 0.14 and 3.4 mmol/L for (−)-epicatechin; 2.9, 1.8, 0.61 and 1.1 mmol/L for catechol, respectively, while the corresponding V_max values were 2.1 × 10⁻², 2.3 × 10⁻², 0.65 × 10⁻² and 0.71 × 10⁻² δA/μg protein min); 1.8 × 10⁻², 0.88 × 10⁻², 0.19 × 10⁻² and 1.0 × 10⁻² δA/μg protein min); 0.48 × 10⁻², 0.59 × 10⁻², 0.67 × 10⁻² and 0.54 × 10⁻² δA/μg protein min), respectively. FT-IR indicated the formation of probable dimer from (+)-catechin in organic solvent. These results suggest that this laccase has higher catalytic oxidation capacity of phenolic compounds in suitable organic media and favorite oligomers could be obtained.