Human-associated Staphylococcus aureus strains within great ape populations in Central Africa (Gabon)

The risk of serious infections caused by Staphylococcus aureus is well-known. However, most studies regarding the distribution of (clinically relevant) S. aureus among humans and animals took place in the western hemisphere and only limited data are available from (Central) Africa. In this context, recent studies focused on S. aureus strains in humans and primates, but the question of whether humans and monkeys share related S. aureus strains or may interchange strains remained largely unsolved. In this study we aimed to evaluate the distribution and spread of human-like S. aureus strains among great apes living in captivity. Therefore, a primate facility at the International Centre for Medical Research of Franceville (Gabon) was screened. We detected among the primates a common human S. aureus strain, belonging to the spa-type t148. It was isolated from three different individuals of the western lowland gorilla (Gorilla gorilla gorilla), of which one individual showed a large necrotizing wound. This animal died, most probably of a staphylococcal sepsis. Additionally, we discovered the t148 type among chimpanzees (Pan troglodytes) that were settled in the immediate neighbourhood of the infected gorillas. A detailed analysis by pulsed field gel electrophoresis showed that the gorilla and chimpanzee isolates represented two closely related strains. To our knowledge, this is the first report of a human-associated S. aureus strain causing disease in great apes. The simultaneous detection in gorillas and chimpanzees indicated an interspecies transmission of this S. aureus strain. Our results recommend that protection of wild animals must not only be based on habitat conservation, but also on the assessment of the risk of contact with human pathogens.     

Detection and genotyping of human papillomavirus by real-time PCR assay

BackgroundDiagnosis of human papillomavirus (HPV) disease remains a challenge due to several factors related to the cost, the workload of available commercial assays to detect and genotype HPV, and to the low prevalence of infected patients.ObjectiveOur study aimed to develop a real-time PCR, based on SPF10 primers, in order to combine HPV-DNA detection and genotype identification avoiding the negative samples.Study designValidation of SYBR-green based SPF10 real-time PCR on HPV-DNA plasmids followed by the investigation of the viral status in 92 samples from oropharyngeal (94%) cutaneous biopsies (3%) and anal smears (3%) which had previously been HPV-genotyped by LiPA hybridization. In-house HPV viral loads were performed to evaluate the SPF10 real-time PCR sensitivity.ResultsData showed that 100% of HPV plasmids, assessable by LiPA hybridization, were detected and genotyped appropriately after SPF10 real-time PCR assays. These results defined a range of melting temperature peaks for HPV positivity by real-time PCR. The efficient determination of the presence of HPV-DNA by SPF10 real-time PCR was validated for 98% of clinical samples compared to commercial method. Discordant results were due to a low HPV-DNA amount and to a supplementary HPV genotype identified. The SPF10 real-time PCR sensitivity was evaluated between 1 and 10 copies/10³ cells using in-house HPV (6, 11 and 16) viral load assays.ConclusionThe real-time PCR method was efficient in combining screening and genotyping of HPV-DNA. Cost and workload reduction by SPF10 real-time PCR approach may facilitate earlier diagnosis and clinical management of HPV infected patients.     

Down-Regulated CK8 Expression in Human Intervertebral Disc Degeneration

As an intermediate filament protein, cytokeratin 8 (CK8) exerts multiple cellular functions. Moreover, it has been identified as a marker of notochord cells, which play essential roles in human nucleus pulposus (NP). However, the distribution of CK8 positive cells in human NP and their relationship with intervertebral disc degeneration (IDD) have not been clarified until now. Here, we found the percentage of CK8 positive cells in IDD (25.7±4.14%) was significantly lower than that in normal and scoliosis NP (51.9±9.73% and 47.8±5.51%, respectively, p<0.05). Western blotting and qRT-PCR results confirmed the down-regulation of CK8 expression in IDD on both of protein and mRNA levels. Furthermore, approximately 37.4% of cell clusters were CK8 positive in IDD. Taken together, this is the first study to show a down-regulated CK8 expression and the percentage of CK8 positive cell clusters in IDD based upon multiple lines of evidence. Consequently, CK8 positive cells might be considered as a potential option in the development of cellular treatment strategies for NP repair.     

Lentivirus-based RNA Silencing of Nemo-like Kinase (NLK) Inhibits the CAL 27 Human Adenosquamos Carcinoma Cells Proliferation and Blocks G0/G1 Phase to S Phase

Background: The Nemo-like kinase (NLK) is a serine/threonine-protein kinase that involved in a number of signaling pathways regulating cell fate. Variation of NLK has been shown to be associated with the risk of cancer. However, the function of NLK in oral adenosquamous carcinoma cells line CAL-27 is unknown.Methods: In this study, we evaluated the function of NLK in CAL-27 cells by using lentivirus-mediated RNA silence. The targeted gene expression, cell proliferation and cell cycle are investigated by RT-PCR, western-blot, MTT method, colony forming assay and flow cytometry analysis respectively.Results: After NLK silencing, the number of colonies was significantly reduced (54±5 colonies/well compared with 262±18 colonies/well in non-infected or 226±4 colonies/well in negative control group (sequence not related to NLK sequence with mismatched bases). Using crystal violet staining, we also found that the cell number per colony was dramatically reduced. The RNA silencing of NLK blocks the G0/G1 phase to S phase progression during the cell cycle.Conclusions: These results suggest that NLK silencing by lentivirus-mediated RNA interference would be a potential therapeutic method to control oral squamous carcinoma growth.     

Vascular morphology and blood flow signatures for differential artery-vein analysis in optical coherence tomography of the retina

Differential artery-vein (AV) analysis is essential for retinal study, disease detection, and treatment assessment. This study is to characterize vascular reflectance profiles and blood flow patterns of retinal artery and vein systems in optical coherence tomography (OCT) and OCT angiography (OCTA), and establish them as robust signatures for objective AV classification. A custom designed OCT was employed for three-dimensional (3D) imaging of mouse retina, and corresponding OCTA was reconstructed. Radially resliced OCT B-scans revealed two, i.e. top and bottom, hyperreflective wall boundaries in retinal arteries, while these wall boundaries were absent in OCT of retinal veins. Additional OCTA analysis consistently displayed a layered speckle distribution in the vein, which may indicate the venous laminar flow. These OCT and OCTA differences offer unique signatures for objective AV classification in OCT and OCTA.     

Xenogeneic bone filling materials modulate mesenchymal stem cell recruitment: role of the Complement C5a

When bone filling materials are applied onto the periodontal tissues in vivo, they interact with the injured periodontal ligament (PDL) tissue and modulate     

基于UPLC-Q-TOF-MS技术分析乌蕨中的化学成分

目的：快速识别乌蕨中化学成分。方法：利用UPLC-Q-TOF-MS/MS技术对乌蕨药材进行分析,采用Waters C₁₈色谱柱,流动相为乙腈-0.2%甲酸溶液,流速为0.25 ml/min,柱温40℃;采用正、负离子模式进行检测;利用PeakView 1.2软件对所得的成分进行分析,整理其母离子,及二级碎片,通过与对照品、相关文献比对,分析其裂解途径,推断其化合物结构。结果：共推断出乌蕨中32种化合物,包括15个黄酮类化合物,15个有机酸类化合物,2个香豆素类成分,其中原儿茶酸葡萄糖苷、咖啡酰葡萄糖苷首次从乌蕨中发现。结论：从液质结果及相关文献表明,乌蕨中以有机酸、黄酮及其苷类成分为主。本研究可为乌蕨的药效物质基础研究提供参考依据。     

联合治疗方案对晚期非小细胞肺癌患者预后及免疫功能的影响

目的探讨树突状细胞-细胞因子诱导杀伤细胞(DC-CIK)过继免疫治疗及多西他赛联合顺铂(DP)化疗方案对晚期非小细胞肺癌患者预后及免疫功能的影响。方法选取60例晚期非小细胞肺癌患者为研究对象,随机分为DP化疗组(化疗组,n=30)及DC-CIK细胞过继免疫治疗联合DP化疗组(联合组,n=30)。2个疗程后,评估2组患者治疗效果、免疫功能以及不良反应发生情况。采用Kaplan-Meier比较2组患者生存情况。结果治疗后,联合组客观有效率(ORR)及疾病控制率(DCR)均高于化疗组,差异有统计学意义(P 0.05)。治疗后,2组外周血免疫球蛋白G(IgG)、免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、CD3~+、CD8~+、自然杀伤细胞(NK)水平均高于治疗前和化疗组,差异有统计学意义(P 0.05); 2组血清CD4~+水平以及CD4~+/CD8~+值均低于治疗前,且联合组以上指标低于化疗组,差异有统计学意义(P 0.05)。2组不良反应发生率比较,差异无统计学意义(P0.05)。化疗组患者中位无进展生存期、中位总生存期分别为3.33、6.37个月,联合组分别为6.40、8.40个月。Kaplan-Meier生存分析结果显示,联合组患者生存情况优于化疗组。结论 DC-CIK细胞过继免疫治疗联合DP化疗方案有利于提高晚期非小细胞肺癌患者的治疗效果,改善免疫功能,延长生存期,且不增加不良反应发生风险。     

BDNF基因多态性对恐惧记忆获得和消退的影响研究

目的:探索脑源性神经营养因子(BDNF)rs6265位点基因多态性与条件性恐惧记忆获得、消退及复燃的关系。方法:招募符合入组和排除标准的90名18-30岁的汉族健康男性,采集人口学资料、血液样本并进行恐惧记忆的学习、消退及测试。采用MassARRAY基因分型方法确定受试者在BDNF基因位点rs6265的单核苷酸多态性,采用方差分析探索BDNF rs6265位点与恐惧记忆学习和消退的关联。结果:BDNF rs6265位点的三种基因型CC、CT、TT对恐惧记忆获得、消退、自发恢复和复燃的影响无显著差异(P>0.05)。结论:BDNF rs6265多态性位点不影响恐惧记忆的获得、消退和表达。