Compositions of microbial communities associated with oil and water in a mesothermic oil field

Samples of produced water and oil obtained from the Enermark field (near Medicine Hat, Alberta, Canada) were separated into oil and aqueous phases first gravitationally and then through centrifugation at 20°C in an atmosphere of 90% N₂ and 10% CO₂. Biomass that remained associated with oil after gravitational separation (1×g) was dislodged by centrifugation at 25,000×g. DNA was isolated from the aqueous and oil-associated biomass fractions and subjected to polymerase chain reaction amplification with primers targeting bacterial and archaeal 16S rRNA genes. DNA pyrosequencing and bioinformatics tools were used to characterize the resulting 16S rRNA gene amplicons. The oil-associated microbial community was less diverse than that of the aqueous phase and had consistently higher representation of hydrogenotrophs (methanogens of the genera Methanolobus and Methanobacterium and acetogens of the genus Acetobacterium), indicating the oil phase to be a primary source of hydrogen. Many known hydrocarbon degraders were also found to be oil-attached, e.g. representatives of the gammaproteobacterial genus Thalassolituus, the actinobacterial genus Rhodococcus and the alphaproteobacterial genera Sphingomonas, Brevundimonas and Stappia. In contrast, all eight representatives of genera of the Deltaproteobacteria identified were found to be associated with the aqueous phase, likely because their preferred growth substrates are mostly water-soluble. Hence, oil attachment was seen for genera acting on substrates found primarily in the oil phase.     

The detection of hTERC amplification using fluorescence in situ hybridization in the diagnosis and prognosis of cervical intraepithelial neoplasia: a case control study

ABSTRACT: BACKGROUND: Currently the routine non-invasive screening methods for cervical intraepithelial neoplasia (CIN) and cervical cancer are Thinprep cytology test (TCT) and human papillomavirus testing. However, both methods are limited by the high false positive and false negative rates and lack of association with patients' prognosis, especially for the early detection of pro-malignant CIN. The aim of the study was to investigate the role of genomic amplification of human telomerase gene (hTERC) in the diagnosis and prognosis of CIN. METHODS: The study group consisted of specimens of exfoliated cervical cells from 151 patients, including 27 with CIN I, 54 with CIN II/III, 17 with carcinoma in situ, and 28 with invasive squamous carcinoma, as well as 25 patients who were at 2-year follow-up after either Loop Electrosurgical Excision treatment (n = 11) or radical surgery (n = 14). hTERC amplification was detected by dual-color interphase fluorescence in situ hybridization (FISH), and the results were compared with TCT and histologic examination. The final diagnosis was determined by the pathological examination. The control group consisted of specimens of exfoliated cervical cells from 40 normal women. RESULTS: The percentage of cervical exfoliated cells with positive hTERC amplification and incidence rates of hTERC amplification were 9.2% [PLUS-MINUS SIGN] 4.6% and 44.4% (12/27) respectively in patients with CIN I; 16.0% [PLUS-MINUS SIGN] 14.4% and 85.1% (46/54) in patients with CIN II/III; 19.7% [PLUS-MINUS SIGN] 13.3% and 88.3% (15 /17) in patients with carcinoma in situ; 47.0% [PLUS-MINUS SIGN] 25.2% and 100% (28/28)in patients with invasive squamous carcinoma. There was statistically significant difference between the control and study group (P <0.01), and between the patients with various diseases within the study group (P <0.05). CONCLUSION: The detection of genomic amplification of hTERC using FISH is a non-invasive and effective approach for CIN.     

Single collateral reconstructions reveal distinct phases of corticospinal remodeling after spinal cord injury

Background

Injuries to the spinal cord often result in severe functional deficits that, in case of incomplete injuries, can be partially compensated by axonal remodeling. The corticospinal tract (CST), for example, responds to a thoracic transection with the formation of an intraspinal detour circuit. The key step for the formation of the detour circuit is the sprouting of new CST collaterals in the cervical spinal cord that contact local interneurons. How individual collaterals are formed and refined over time is incompletely understood.

Methodology/Principal Findings

We traced the hindlimb corticospinal tract at different timepoints after lesion to show that cervical collateral formation is initiated in the first 10 days. These collaterals can then persist for at least 24 weeks. Interestingly, both major and minor CST components contribute to the formation of persistent CST collaterals. We then developed an approach to label single CST collaterals based on viral gene transfer of the Cre recombinase to a small number of cortical projection neurons in Thy1-STP-YFP or Thy1-Brainbow mice. Reconstruction and analysis of single collaterals for up to 12 weeks after lesion revealed that CST remodeling evolves in 3 phases. Collateral growth is initiated in the first 10 days after lesion. Between 10 days and 3–4 weeks after lesion elongated and highly branched collaterals form in the gray matter, the complexity of which depends on the CST component they originate from. Finally, between 3–4 weeks and 12 weeks after lesion the size of CST collaterals remains largely unchanged, while the pattern of their contacts onto interneurons matures.

Conclusions/Significance

This study provides a comprehensive anatomical analysis of CST reorganization after injury and reveals that CST remodeling occurs in distinct phases. Our results and techniques should facilitate future efforts to unravel the mechanisms that govern CST remodeling and to promote functional recovery after spinal cord injury.     

Smo基因在人肝癌Huh-7细胞中的表达及小RNA干扰Smo基因表达对肝癌Huh-7细胞增殖及凋亡的影响

目的:在细胞学层面上研究Smo基因在人肝癌Huh-7细胞中的表达及小RNA干扰Smo基因表达对肝癌Huh-7细胞增殖及凋亡的影响。方法:Huh-7细胞培养,总RNA抽提,紫外分光光度计纯度测定,Western印记法检测Smo蛋白表达,转染后流式细胞检测Huh-7凋亡率。结果:在mRNA和蛋白水平Smo均强表达。siRNA-1干扰序列干扰结果最强,转染后可诱导Huh-7细胞凋亡。结论:siRNA-l能对肝癌Huh7细胞Smo基因表达产生干涉作用,siRNA-1序列能有效地降解肝癌Huh7细胞内的SmomRNA,使Smo mRNA及Smo蛋白表达下调,从而达到沉默肝癌Huh7细胞中Smo mRNA表达的效果。     

疑难全血细胞减少症的免疫学变化及病因分析

目的:研究疑难全血细胞减少症的免疫学指标改变,探讨其病因构成及预后.并研究骨髓Coombs试验在疑难全血细胞减少症病因诊断中的意义.方法:对36例疑难全血细胞减少症患者的免疫指标,T细胞亚群,骨髓Coombs试验结果进行分析,根据免疫指标的变化,给予部分患者试验性用药,出院后随访一年.另以25例再生障碍性贫血患者为再障对照组,以22例健康成人为健康对照组,比较骨髓Coombs试验阳性率的差异,X2检验P＜0.05有统计学意义.结果:36位患者,18例诊断明确:5例为再生障碍性贫血,6例为免疫相关性全血细胞减少症,3例为系统性红斑狼疮,1例骨髓纤维化,1例肝癌,2例与妊娠相关的全血细胞减少;18例诊断不明确:8例患者于试验性升血治疗后血象好转,2例好转不明显,8例失访.骨髓Coombs试验阳性率在研究组与对照组的比较:疑难全血细胞减少症组(16.7％)高于再障对照组(X2=4.621,P＜0.05),也高于正常对照组(x2□4.09,P＜0.05)差异有统计学意义.结论:1对疑难全血细胞减少症患者进行试验性激素治疗及长期随访,有利于疾病的最终诊断.2免疫指标的检查及动态监测对明确疑难全血细胞减少症的病因十分重要.3骨髓Coombs试验对明确疑难全血细胞减少症病因有重要意义.     

万古霉素- 脂质体对兔股骨头置换感染模型预防作用的实验研究

目的:制备万古霉素脂质体抗生素缓释系统,研究该缓释系统在治疗兔人工股骨头置换术后感染中的作用,为临床的进一步应用提供理论基础和实验依据。方法 :建立兔人工股骨头置换术后感染模型,将配置好的万古霉素-脂质体药物释放系统、万骨霉素于术中即分别放入关节腔及髓腔内,同时设对照组。大体观察左侧髋关节及观察右髋关节的活动度。实验动物于人工股骨头置换术后即刻、第4、8周,分别行髋关节X线摄片检查。实验动物建立感染模型6周后,分别取关节腔内肉芽组织或假膜组织,观察细菌生长情况,计数细菌菌落。并用4%的甲醛溶液固定组织,石蜡包埋(骨组织先用EDTA脱钙处理),HE染色,光镜观察组织炎细胞浸润状况。实验动物建立感染模型术前和术后第1、3、6周后,测定血清C-反应蛋白和血沉的变化。结果:成功建立了兔人工股骨头置换术后感染模型,各组兔人工股骨头置换术后股骨头均在位,经万古霉素-脂质体药物释放系统治疗后,万古霉素-脂质体组翻修术后8周X线结果提示关节间隙清晰,无明显骨缺损即透亮线形成,而万古霉素-骨水泥组置换术前后对比片,无明显变化。对照组感染未控制者术后6周X线结果显示,关节间隙模糊,假体及股骨结合部出现轻度骨吸收。细菌培养结果显示,空白对照组中实验动物细菌感染率为100%,而加入万古霉素后,万古霉素-PMMA组的实验动物细菌感染率为33.3%,显著降低;万古霉素-脂质体组实验动物细菌感染率为16.7%,与万古霉素-PMMA组相比,感染率显著降低(P<0.05);联合应用万古霉素-PMMA和万古霉素-脂质体,与其它组相比较,感染率显著降低(P<0.05),提示这种模式具有良好的抑菌效果。病理组织切片,发现实验动物建立感染模型6周后,空白对照组中实验动物感染人工股骨头周围股骨组织明显充血及炎性细胞浸润,万古霉素-PMMA组、万古霉素-脂质体组和万古霉素-脂质体组+万古霉素-PMMA的实验动物,炎性细胞浸润明显减少,提示术后关节周围细菌感染得到了有效的抑制。与术前相比较,各组CRP均显著升高(P<0.01),而经过万古霉素-PMMA组、万古霉素-脂质体组和万古霉素-脂质体组+万古霉素-PMMA治疗的实验动物,CRP均显著低于空白对照组中实验动物(P<0.05)。结论:本实验中首先建立兔人工股骨头置换术后感染模型,并观察了万古霉素-脂质体药物释放系统对兔人工股骨头置换术后感染的效应,通过X线检查、细菌培养、病理组织切片观察以及血清C-反应蛋白和血沉变化的测定,结果表明,通过上述方法制备的万古霉素脂质体对于人工股骨头置换术后感染具有良好的预防和治疗作用,为临床的进一步应用提供了初步的实验基础和理论依据。     

骨髓间充质干细胞和内皮祖细胞移植促进血流重建的比较

目的:比较骨髓间充质细胞(Bone Marrow Mesenchymal Stem Cells,BM/MSC)和骨髓源内皮祖细胞(Bone Marrow Endothelialprogenitor cells,BM/EPC)移植促进血流重建的效果,为进一步优化骨髓干细胞移植治疗肢体缺血提供理论基础。方法:获取Lewis大鼠骨髓单个核细胞,在体外培养分化为MSC和EPC。采用Lewis大鼠建立单侧后肢缺血模型。在模型建立后3天,将0.8mlD-Hanks液注入大鼠缺血侧后肢,为对照组(n=6);将8×106个骨髓MSC植入大鼠缺血侧后肢,为MSC组(n=6);将体外培养的8×106个EPC植入大鼠缺血侧后肢,为EPC组(n=6)。细胞移植后3周行缺血大鼠后肢动脉造影,检测缺血侧后肢侧支血管数;获取缺血侧后肢腓肠肌,分别行CD31和α-SMA免疫组化染色,计算毛细血管密度和小动脉密度。结果:MSC组与EPC组侧支血管数无显著性差异,二者均高于对照组;EPC组毛细血管密度明显高于MSC组,二者均高于对照组;MSC组与EPC组小动脉密度无显著性差异,二者均高于对照组。结论:骨髓间充质干细胞移植和内皮祖细胞移植均能够明显促进血流重建,而且骨髓间充质干细胞在治疗肢体缺血性疾病中的优势应该受到重视。     

MRI 在宫颈癌分期中的应用价值

目的:探讨宫颈癌的MRI表现与分期,评价其诊断价值及临床意义。方法:由2名经验丰富的妇科肿瘤医师前瞻性地对80例宫颈活组织病理检查证实为宫颈癌或可疑早期浸润癌的患者进行妇科盆腔检查,共同决定分期和手术可行性。对可直接手术或经先期治疗后有手术可能的患者,于开始治疗前2周内(平均6.5d)行盆腔和腹膜后MRI扫描。在MRI图像上观察原发肿瘤的位置、信号特征及侵犯范围。将MRI分期与临床分期进行对比,手术病理分期为金标准,采用诊断检验方法,以敏感度、特异度、准确度3项指标分析MRI判断宫颈癌分期、宫旁侵犯和盆腔淋巴结转移的价值。结果:MRI对宫颈癌术前分期的总准确度为88.7%,对浸润性癌(Ⅱa及Ⅱa以上)诊断准确度为86.7%。MRI对宫旁侵犯的判断敏感度为85.2%,特异度为85%,准确性为85.1%,MRI预测淋巴结转移的敏感度为58%,特异度为96%,准确性为90%。结论:MRI能多方位清晰显示宫颈癌瘤灶及侵犯范围与途径,明显优于其他影像学检查方法,MRI对术前宫颈癌分期明显优于临床,可做为宫颈癌术前常规的影像检查方法。     

C2C12 细胞胰岛素增敏的机制

目的:观察口服葡萄糖负荷对小鼠小肠组织网膜素基因表达的影响及网膜素对C2C12肌管细胞胰岛素敏感性的影响,并进一步探讨其机制。方法:半定量逆转录聚合酶链反应(RT-PCR)技术检测小鼠小肠组织网膜素mRNA的表达;葡萄糖转运实验观察网膜素对C2C12肌管细胞胰岛素敏感性的影响;Western blot检测Akt(Ser473)的磷酸化水平。结果:口服葡萄糖负荷30min后小鼠小肠组织网膜素基因表达显著减低(P<0.05),而60min后恢复至负荷前水平;葡萄糖转运实验发现:网膜素作用10min对C2C12肌管细胞基础葡萄糖转运无影响,但显著增加了胰岛素刺激的葡萄糖转运(P<0.05);Western Blot发现:网膜素和AICAR均显著增加了C2C12肌管细胞Ak(tSer473)的磷酸化水平(P<0.05)。结论:网膜素作为一种胃肠激素还受到葡萄糖负荷的调节,在C2C12肌管细胞,网膜素可通过增加Akt的磷酸化发挥胰岛素增敏作用。     

MUC3A promotes non-small cell lung cancer progression via activating the NFκB pathway and attenuates radiosensitivity

Mucin 3A (MUC3A) is highly expressed in non-small cell lung cancer (NSCLC), but its functions and effects on clinical outcomes are not well understood. Tissue microarray of 92 NSCLC samples indicated that high levels of MUC3A were associated with poor prognosis, advanced staging, and low differentiation. MUC3A knockdown significantly suppressed NSCLC cell proliferation and induced G1/S accumulation via downregulating cell cycle checkpoints. MUC3A knockdown also inhibited tumor growth in vivo and had synergistic effects with radiation. MUC3A knockdown increased radiation-induced DNA double strain breaks and γ-H2AX phosphorylation in NSCLC cells. MUC3A downregulation inhibited the BRCA-1/RAD51 pathway and nucleus translocation of P53 and XCRR6, suggesting that MUC3A promoted DNA damage repair and attenuated radiation sensitivity. MUC3A knockdown also resulted in less nucleus translocation of RELA and P53 in vivo. Immunoprecipitation revealed that MUC3A interacted with RELA and activated the NFκB pathway via promoting RELA phosphorylation and interfering the binding of RELA to IκB. Our studies indicated that MUC3A was a potential oncogene and associated with unfavorable clinical outcomes. NSCLC patients with a high MUC3A level, who should be more frequent follow-up and might benefit less from radiotherapy.