FAS/FASL Expression Profile as a Prognostic Marker in Squamous Cell Carcinoma of the Oral Cavity

FAS/FASL altered expression may cause tumor protecting immunomodulation, with a direct impact on patient prognosis. FAS expression was studied in 60 squamous cell carcinomas of the oral cavity. FAS expression did not show a significant association with tumor histopathological characteristics, but was significantly associated with lymph node positivity. FAS expression was significantly associated with disease specific death and negative FAS expression was an independent risk factor, increasing risk 4 times when compared to positive expression. When FAS and FASL expression results were combined, we were able to define high, intermediate and low risk profiles. Disease-free and disease-specific survival were significantly correlated with FAS/FASL expression profiles. The high risk category was an independent marker for earlier disease relapse and disease-specific death, with approximately 4- and 6-fold increased risk, respectively, when compared to the low risk profile. Risk profiles based on FAS/FASL expression showed that high risk was significantly associated with increased disease relapse and death, as well as shorter disease-free or disease-specific survival. This categorization, added to patient clinical data, may facilitate the choice of therapy, minimizing treatment failure and increasing disease control.     

The effect of rotative stress on CAII,FAS, FASL,OSCAR, and TRAP gene expression in osteoclasts

This study was designed to explore the effects of rotative stress on carbonic anhydrase II (CAII), TNF receptor superfamily member 6 (FAS), FAS ligand (FASL), osteoclast‐associated receptor (OSCAR), and tartrate‐resistant acid phosphatase (TRAP) gene expression in osteoclasts. Osteoclasts were induced from RAW264.7 cells cultured in medium containing recombinant murine soluble receptor activator of NF‐Kβ ligand (sRANKL). The mRNA and protein expression of CAII, FAS, FASL, OSCAR, and TRAP genes in osteoclasts was detected by RT‐PCR and Western blot, respectively, after osteoclasts were loaded at various rotative stress strengths and times. No significant differences in mRNA and protein expression were observed between any of the control groups (P > 0.05). Importantly, rotative stress had a significant effect on the mRNA and protein expression of these genes (P < 0.05). We found a negative relationship between rotative stress strength and prolonged loading time and the expression of FAS/FASL genes in osteoclasts. In addition, there was a positive relationship between rotative stress strength and prolonged loading time and the expression of CAII, OSCAR, or TRAP genes in osteoclasts. Based on these results, rotative stress has a significant effect on CAII, FAS, FASL, OSCAR, and TRAP gene expression in osteoclasts. J. Cell. Biochem. 114: 388–397, 2013. © 2012 Wiley Periodicals, Inc.     

乳酸在中枢神经系统中的作用及其与相关疾病的关系

一直以来,乳酸在脑中被视作代谢废物,对其功能认识严重滞后。近年来,越来越多的证据表明,乳酸在多种生理与病理过程中扮演重要角色。在神经细胞中,星形胶质细胞是产生和释放乳酸的主要细胞源,该细胞通过有氧糖酵解过程生成乳酸,随后经跨膜通道释放至胞外进入神经元为其供能。在中枢神经系统中,乳酸对稳态调节发挥着十分重要的作用。乳酸主要通过两种途径,即代谢途径(作为能量底物)与信号途径(作为信号分子)调控神经元的功能活动,广泛参与神经元能量代谢、兴奋性、可塑性、学习记忆及神经系统发育等生理过程调节,亦参与抑郁行为、阿尔兹海默病(AD)和脑损伤等病理过程的调节。在脑组织中,存在着乳酸特异性受体(GPR81),乳酸与其结合后调控胞内的第二信使。此外,还发现乳酸可通过未知受体调节神经元的兴奋性以及作为信号分子的其他作用。本文就乳酸作为能量底物和信号分子及其参与相关神经疾病的研究进展进行阐述,旨在为相关中枢神经系统疾病防治提供新思路。     

Lactoferrin inhibits dexamethasone-induced chondrocyte impairment from osteoarthritic cartilage through up-regulation of extracellular signal-regulated kinase 1/2 and suppression of FASL,FAS, and Caspase 3

Dexamethasone (Dex) is commonly used for osteoarthritis (OA) with excellent anti-inflammatory and analgesic effect. However, Dex also has many side effects following repeated use over prolonged periods mainly through increasing apoptosis and inhibiting proliferation. Lactoferrin (LF) exerts significantly anabolic effect on many cells and little is known about its effect on OA chondrocytes. Therefore, the aim of this study is to investigate whether LF can inhibit Dex-induced OA chondrocytes apoptosis and explore its possible molecular mechanism involved in. MTT assay was used to determine the optimal concentration of Dex and recombinant human LF (rhLF) on chondrocytes at different time and dose points. Chondrocytes were then stimulated with Dex in the absence or presence of optimal concentration of rhLF. Cell proliferation and viability were evaluated using MTT and LIVE/DEAD assay, respectively. Cell apoptosis was evaluated by multi-parameter apoptosis assay kit using both confocal microscopy and flow cytometry, respectively. The expression of extracellular signal-regulated kinase (ERK), FAS, FASL, and Caspase-3 (CASP3) at the mRNA and protein levels were examined by real-time polymerase chain reaction (PCR) and immunocytochemistry, respectively. The optimal concentration of Dex (25 μg/ml) and rhLF (200 μg/ml) were chosen for the following experiments. rhLF significantly reversed the detrimental effect of Dex on chondrocytes proliferation, viability, and apoptosis. In addition, rhLF significantly prevented Dex-induced down-regulation of ERK and up-regulation of FAS, FASL, and CASP3. These findings demonstrated that rhLF acts as an anabolic effect on chondrocytes through significantly reversing Dex-induced chondrocytes apoptosis. This study may contribute to further investigating the clinical application of LF on OA.     

蚓激酶对人瘢痕疙瘩成纤维细胞的生物学功能和MMP2、MMP9表达的影响

目的:考察蚓激酶对人瘢痕疙瘩成纤维细胞(KF)生物学功能影响,初步阐明其防治瘢痕疙瘩作用机制。方法:选取人KF为体外实验模型,随机分为对照组和蚓激酶低、中、高剂量组;应用药物干预48 h后,采用CCK8法测定人KF增殖率,流式细胞术检测凋亡率,划痕法测迁移能力,Western blot检测MMP2、MMP9蛋白表达。结果:与对照组相比,应用5 U·mL~(-1)、10 U·mL~(-1)和20U·mL~(-1)蚓激酶溶液干预后,人KF增殖率明显降低,凋亡率显著升高,在一定范围内与剂量成正相关性;与对照组相比,蚓激酶组的划痕距离均有不同程度地增加,MMP2、MMP9表达也显著下降,均有显著性差异。结论:蚓激酶可调节人KF的生物学功能,通过抑制细胞增殖和迁移、诱导凋亡、下调迁移相关蛋白MMP2、MMP9表达,防止瘢痕疙瘩的生长和侵润,为蚓激酶的进一步开发和利用提供有力支持。     

Matrilysin/matrix metalloproteinase-7(MMP7) cleavage of perlecan/HSPG2 creates a molecular switch to alter prostate cancer cell behavior

Perlecan/HSPG2, a large heparan sulfate (HS) proteoglycan, normally is expressed in the basement membrane (BM) underlying epithelial and endothelial cells. During prostate cancer (PCa) cell invasion, a variety of proteolytic enzymes are expressed that digest BM components including perlecan. An enzyme upregulated in invasive PCa cells, matrilysin/matrix metalloproteinase-7 (MMP-7), was examined as a candidate for perlecan proteolysis both in silico and in vitro. Purified perlecan showed high sensitivity to MMP-7 digestion even when fully decorated with HS or when presented in native context connected with other BM proteins. In both conditions, MMP-7 produced discrete perlecan fragments corresponding to an origin in immunoglobulin (Ig) repeat region domain IV. While not predicted by in silico analysis, MMP-7 cleaved every subpart of recombinantly generated perlecan domain IV. Other enzymes relevant to PCa that were tested had limited ability to cleave perlecan including prostate specific antigen, hepsin, or fibroblast activation protein α. A long C-terminal portion of perlecan domain IV, Dm IV-3, induced a strong clustering phenotype in the metastatic PCa cell lines, PC-3 and C4-2. MMP-7 digestion of Dm IV-3 reverses the clustering effect into one favoring cell dispersion. In a C4-2 Transwell® invasion assay, perlecan-rich human BM extract that was pre-digested with MMP-7 showed loss of barrier function and permitted a greater level of cell penetration than untreated BM extract. We conclude that enzymatic processing of perlecan in the BM or territorial matrix by MMP-7 as occurs in the invasive tumor microenvironment acts as a molecular switch to alter PCa cell behavior and favor cell dispersion and invasiveness.     

Smad7对肝癌细胞增殖和迁移的影响及其临床意义

目的:探究Smad7对肝癌细胞增殖和迁移的影响及其临床意义。方法:通过转染pcDNA3.1(+)-Smad7质粒或Smad7的小干扰RNA使得Smad7在肝癌细胞系HepG2和Huh7中过表达或敲减,应用MTT法检测Smad7对肝癌细胞增殖的影响,采用细胞划痕实验以及Transwell实验探讨Smad7对肝癌细胞迁移的影响。采用qRT-PCR检测9例肝癌癌患者手术切除的组织样本中Smad7的表达。结果:过表达Smad7的肝癌细胞增殖能力与对照组相比有明显的下降,而敲减smad7能够促进肝癌细胞的增殖。过表达smad7的肝癌细胞穿过Transwell小室底膜的能力显著下降,而敲减Smad7能够促进这种能力。Smad7在肝癌癌旁组织中的表达显著高于癌组织。结论:Smad7能够在肝细胞肝癌的进展中发挥负向调控作用。     

Vpu Exploits the Cross-Talk between BST2 and the ILT7 Receptor to Suppress Anti-HIV-1 Responses by Plasmacytoid Dendritic Cells

Plasmacytoid dendritic cells (pDCs) constitute a major source of type-I interferon (IFN-I) production during acute HIV infection. Their activation results primarily from TLR7-mediated sensing of HIV-infected cells. However, the interactions between HIV-infected T cells and pDCs that modulate this sensing process remain poorly understood. BST2/Tetherin is a restriction factor that inhibits HIV release by cross-linking virions onto infected cell surface. BST2 was also shown to engage the ILT7 pDC-specific inhibitory receptor and repress TLR7/9-mediated IFN-I production by activated pDCs. Here, we show that Vpu, the HIV-1 antagonist of BST2, suppresses TLR7-mediated IFN-I production by pDC through a mechanism that relies on the interaction of BST2 on HIV-producing cells with ILT7. Even though Vpu downregulates surface BST2 as a mean to counteract the restriction on HIV-1 release, we also find that the viral protein re-locates remaining BST2 molecules outside viral assembly sites where they are free to bind and activate ILT7 upon cell-to-cell contact. This study shows that through a targeted regulation of surface BST2, Vpu promotes HIV-1 release and limits pDC antiviral responses upon sensing of infected cells. This mechanism of innate immune evasion is likely to be important for an efficient early viral dissemination during acute infection.     

Role of Pax3/7 in the tectum regionalization

Pax3/7 is expressed in the alar plate of the mesencephalon. The optic tectum differentiates from the alar plate of the mesencephalon, and expression of Pax3/7 is well correlated to the tectum development. To explore the function of Pax3 and Pax7 in the tectum development, we misexpressed Pax3 and Pax7 in the diencephalon and ventral mesencephalon. Morphological and molecular marker gene analysis indicated that Pax3 and Pax7 misexpression caused fate change of the alar plate of the presumptive diencephalon to that of the mesencephalon, that is, a tectum and a torus semicircularis were formed ectopically. Ectopic tectum in the diencephalon appeared to be generated through sequential induction of Fgf8, En2 and Pax3/7. In ventral mesencephalon, which expresses En but does not differentiate to the tectum in normal development, Pax3 and Pax7 misexpression induced ectopic tectum. In normal development, Pax3 and Pax7 expression in the mesencephalon commences after Otx2, En and Pax2/5 expression. In addition, expression domain of Pax3 and Pax7 is well consistent with presumptive tectum region in a dorsoventral axis. Taken together with normal expression pattern of Pax3 and Pax7, results of misexpression experiments suggest that Pax3 and Pax7 define the tectum region subsequent to the function of Otx2 and En.     

卡培他滨联合奥沙利铂治疗晚期结直肠癌的疗效及对生活质量的影响

目的:探讨卡培他滨联合奥沙利铂治疗晚期结直肠癌患者的临床疗效及对患者生活质量的影响。方法:选取我院2013年3月-2015年12月收治的40例晚期结直肠癌患者,按乱数表法分为观察组和对照组各20例。对照组给予卡培他滨治疗,观察组给予卡培他滨联合奥沙利铂治疗,两组均治疗3周期。对比两组患者治疗后4周的客观缓解率和临床受益率,对比两组患者治疗前、治疗后4周的功能状态评分(KPS)和体力状况评分(ZPS),对比两组患者6个月、1年生存率以及并发症发生率。结果:治疗后4周观察组临床受益率和客观缓解率显著高于对照组,差异有统计学意义(P0.05);与治疗前相比,治疗后4周两组KPS评分显著升高,ZPS评分显著降低,差异均有统计学意义(P0.05);与对照组相比,治疗后4周观察组KPS评分显著升高,ZPS评分显著降低,差异均有统计学意义(P0.05);两组患者在治疗过程中恶心呕吐、口腔黏膜炎、贫血、血小板减少、白细胞减少、腹泻等并发症发生率比较差异均无统计学意义(P0.05);观察组1年生存率显著高于对照组,差异有统计学意义(P0.05)。结论:卡培他滨联合奥沙利铂治疗晚期结直肠癌患者疗效较好,能提高患者的客观缓解率、临床受益率、生活质量和1年生存率,较单用卡培他滨治疗优势明显,值得临床推广。     

The Mechanistic Basis of Myxococcus xanthus Rippling Behavior and Its Physiological Role during Predation

Myxococcus xanthus cells self-organize into periodic bands of traveling waves, termed ripples, during multicellular fruiting body development and predation on other bacteria. To investigate the mechanistic basis of rippling behavior and its physiological role during predation by this Gram-negative soil bacterium, we have used an approach that combines mathematical modeling with experimental observations. Specifically, we developed an agent-based model (ABM) to simulate rippling behavior that employs a new signaling mechanism to trigger cellular reversals. The ABM has demonstrated that three ingredients are sufficient to generate rippling behavior: (i) side-to-side signaling between two cells that causes one of the cells to reverse, (ii) a minimal refractory time period after each reversal during which cells cannot reverse again, and (iii) physical interactions that cause the cells to locally align. To explain why rippling behavior appears as a consequence of the presence of prey, we postulate that prey-associated macromolecules indirectly induce ripples by stimulating side-to-side contact-mediated signaling. In parallel to the simulations, M. xanthus predatory rippling behavior was experimentally observed and analyzed using time-lapse microscopy. A formalized relationship between the wavelength, reversal time, and cell velocity has been predicted by the simulations and confirmed by the experimental data. Furthermore, the results suggest that the physiological role of rippling behavior during M. xanthus predation is to increase the rate of spreading over prey cells due to increased side-to-side contact-mediated signaling and to allow predatory cells to remain on the prey longer as a result of more periodic cell motility.     

MMP14在非小细胞肺癌中的表达及其与预后的关系

肺癌5年生存率低,侵润和转移是导致其死亡的主要原因.探讨MMP14(matrix mettallo proteinase 14)在非小细胞肺癌(non-small cell lung cancer,NSCLC)组织中的表达,及其与浸润转移和预后的关系.通过采用免疫组化检测92例NSCLC组织及25例癌旁正常组织中MMP14蛋白的表达,并分析其与临床病理学特征及预后的关系.免疫组化结果显示,MMP14蛋白在NSCLC组织中表达的阳性率为64.1%(59/92),显著高于癌旁正常组8%(2/25)(P<0.001).MMP14阳性率与NSCLC的分化程度、T分期、淋巴结转移和TNM分期相关(P<0.05),而与性别、年龄、吸烟及组织学类型无关(P>0.05).Kaplan-meier生存曲线显示,MMP14阳性表达组的患者5年生存期显著低于阴性表达组(P=0.004).Cox多因素回归分析显示,MMP14不是NSCLC的独立预后因素(P>0.05).MMP14在肺癌的分化、浸润和转移中扮演着重要的作用,并对生存期有一定的影响.     

MMP11在肺鳞癌和腺癌组织中的表达及与预后的关系

MMP11属于MMPs家族,它与肿瘤的浸润转移密切相关.探讨MMP11在肺鳞癌和腺癌组织中的表达,及其与浸润转移和预后的关系.应用免疫组化检测50例肺鳞癌和38例肺腺癌组织中MMP11蛋白的表达.结果显示,MMP11在肺鳞癌和腺癌组织中的阳性率分别为68.0%(34/50)和73.7%(28/38)(P=0.563).MMP11的阳性率随着肺癌T分期的增加、淋巴结转移和TNM分期的增加而增加(P<0.05).Kaplan-meier单因素生存分析显示,MMP11表达(P=0.001)、T分期(P=0.009)、淋巴结转移(P<0.001)和TNM分期(P<0.001)对生存期的影响有统计学意义.Cox多因素分析显示,只有TNM分期(P=0.028)是肺癌的独立预后危险因素,MMP11(P=0.105)不是肺癌的独立预后危险因素.MMP11的表达与肺癌的浸润、转移及生存期密切相关.     

LncRNA SNHG4在结直肠癌中的表达及对细胞奥沙利铂耐药性的影响

目的:探讨长链非编码RNA(LncRNA)SNHG4在结直肠癌(CRC)中的表达以及对细胞奥沙利铂耐药性的影响。方法:采用实时定量PCR(qRT-PCR)法检测24例CRC组织及其邻近癌旁组织中LncRNA SNHG4的表达水平,并用费希尔精确检验(Fisher's exact test)分析LncRNA SNHG4表达水平与CRC患者临床病理特征相关性。体外培养HCT116细胞,将HCT116细胞分为sh-SNHG4组(敲减组)、sh-NC组(阴性对照组),qRT-PCR法检测各组HCT116细胞中LncRNA SNHG4表达水平,CCK8法检测两组细胞经梯度浓度(1.0、2.5、5.0、10.0、20.0μmol/L)奥沙利铂(L-OHP)处理24小时后细胞增殖能力变化情况。EdU和免疫荧光(Immunofluorescence,IF)实验检测两组细胞经10μmol/L的L-OHP处理24小时后,细胞增殖能力以及核DNA损伤情况。结果:CRC癌组织中LncRNA SNHG4相对表达水平与癌旁组织相比明显升高(P<0.05),其表达水平与CRC患者淋巴结转移、TNM分期、脉管侵犯显著相关(P<0.05)。与sh-NC组相比,sh-SNHG4组在梯度浓度的L-OHP处理下,相对细胞活性下降更加明显(P<0.05)。在10μmol/L的L-OHP处理条件下,sh-SNHG4组相比sh-NC组,细胞增殖能力减弱(P<0.05),核DNA损伤情况更严重(P<0.05)。结论:LncRNA SNHG4在CRC组织中高表达,敲减LncRNA SNHG4可以减弱HCT116细胞对L-OHP耐药性。     

The Approach to Sample Acquisition and Its Impact on the Derived Human Fecal Microbiome and VOC Metabolome

Recent studies have illustrated the importance of the microbiota in maintaining a healthy state, as well as promoting disease states. The intestinal microbiota exerts its effects primarily through its metabolites, and metabolomics investigations have begun to evaluate the diagnostic and health implications of volatile organic compounds (VOCs) isolated from human feces, enabled by specialized sampling methods such as headspace solid-phase microextraction (hSPME). The approach to stool sample collection is an important consideration that could potentially introduce bias and affect the outcome of a fecal metagenomic and metabolomic investigation. To address this concern, a comparison of endoscopically collected (in vivo) and home collected (ex vivo) fecal samples was performed, revealing slight variability in the derived microbiomes. In contrast, the VOC metabolomes differ widely between the home collected and endoscopy collected samples. Additionally, as the VOC extraction profile is hyperbolic, with short extraction durations more vulnerable to variation than extractions continued to equilibrium, a second goal of our investigation was to ascertain if hSPME-based fecal metabolomics studies might be biased by the extraction duration employed. As anticipated, prolonged extraction (18 hours) results in the identification of considerably more metabolites than short (20 minute) extractions. A comparison of the metabolomes reveals several analytes deemed unique to a cohort with the 20 minute extraction, but found common to both cohorts when the VOC extraction was performed for 18 hours. Moreover, numerous analytes perceived to have significant fold change with a 20 minute extraction were found insignificant in fold change with the prolonged extraction, underscoring the potential for bias associated with a 20 minute hSPME.     

Integrins regulate DLG/FAS2 via a CaM kinase II-dependent pathway to mediate synapse elaboration and stabilization during postembryonic development

Calcium/calmodulin dependent kinase II (CaMKII), PDZ-domain scaffolding protein Discs-large (DLG), immunoglobin superfamily cell adhesion molecule Fasciclin 2 (FAS2) and the position specific (PS) integrin receptors, including betaPS and its alpha partners (alphaPS1, alphaPS2, alphaPS3/alphaVolado), are all known to regulate the postembryonic development of synaptic terminal arborization at the Drosophila neuromuscular junction (NMJ). Recent work has shown that DLG and FAS2 function together to modulate activity-dependent synaptic development and that this role is regulated by activation of CaMKII. We show that PS integrins function upstream of CaMKII in the development of synaptic architecture at the NMJ. betaPS integrin physically associates with the synaptic complex anchored by the DLG scaffolding protein, which contains CaMKII and FAS2. We demonstrate an alteration of the FAS2 molecular cascade in integrin regulatory mutants, as a result of CaMKII/integrin interactions. Regulatory betaPS integrin mutations increase the expression and synaptic localization of FAS2. Synaptic structural defects in betaPS integrin mutants are rescued by transgenic overexpression of CaMKII (proximal in pathway) or genetic reduction of FAS2 (distal in pathway). These studies demonstrate that betaPS integrins act through CaMKII activation to control the localization of synaptic proteins involved in the development of NMJ synaptic morphology.