Nondestructive evaluation of photosynthesis by delayed luminescence in Arabidopsis in Petri dishes

Nondestructive evaluation of photosynthesis is a valuable tool in the field and laboratory. Delayed luminescence (DL) can reflect charge recombination through the backflow of electrons. However, DL detection has not yet been adapted for whole plants in Petri dishes. To compensate for differences in DL decay between sibling Arabidopsis plants grown under the same conditions, we developed a time-sequential double measurement method. Using this method, we examined the influence of photosynthetic electron flow inhibitors, and differences in the DL decay curves were categorized by considering the initial and late phases of the decay curves, as well as their intermediate slopes. The appearance of concavity and convexity in DL curves in Arabidopsis was different from unicellular algae, suggesting complexity in the photosynthetic machinery of higher plants. This detection method should be invaluable for evaluating photosynthetic defects in higher plants under sterile conditions without interrupting plant culture.     

Anti-bacterial and anti-inflammatory effects of ethanol extract from Houttuynia cordata poultice

Houttuynia cordata (HC) has been commonly used as many traditional remedies in local areas of Japan. Although many pharmacological activities of HC have been reported, the mechanism underlying the effect of HC remains unknown. We conducted the interview survey in Japan to verify how HC was actually used. The interview survey revealed that HC poultice (HCP) prepared from smothering fresh leaves of HC was most frequently used for the treatment of purulent skin diseases including furuncle and carbuncle with high effectiveness. Ethanol extract of HCP (eHCP) showed anti-bacterial effects against methicillin-resistant Staphylococcus aureus (MRSA), and showed an anti-biofilm activity against MRSA. eHCP showed dose-dependent inhibition of S. aureus lipoteichoic acid (LTA)-induced interleukin-8 and CCL20 production in human keratinocyte without any cytotoxicity. These results suggest that HCP is effective for skin abscess and its underlying mechanism might be the complicated multiple activities for both bacteria and host cells.     

Effects of 8-halo-7-deaza-2′-deoxyguanosine triphosphate on DNA synthesis by DNA polymerases and cell proliferation

8-OxodG (8-oxo-2′-deoxyguanosine) is representative of nucleoside damage and shows a genotoxicity. To significantly reveal the contributions of 7-NH and C8-oxygen to the mutagenic effect of 8-oxodG by DNA polymerases, we evaluated the effects of the 8-halo-7-deaza-dG (8-halogenated 7-deaza-2′-deoxyguanosine) derivatives by DNA polymerases. 8-Halo-7-deaza-dGTPs were poorly incorporated by both KF(exo⁻) and human DNA polymerase β opposite dC or dA into the template DNA. Furthermore, it was found that KF(exo⁻) was very sensitive to the introduction of the C8-halogen, while polymerase β can accommodate the C8-halogen resulting in an efficient dCTP insertion opposite the 8-halo-7-deaza-dG in the template DNA. These results indicate that strong hydrogen bonding between 7-NH in the 8-oxo-G nucleobase and 1-N in the adenine at the active site of the DNA polymerase is required for the mutagenic effects. Whereas, I-deaza-dGTP shows an antiproliferative effect for the HeLa cells, suggesting that it could become a candidate as a new antitumor agent.     

小花草玉梅正常和自然变异植株的AP3-3基因研究

野生小花草玉梅(Anemone rivularis var.flore-minore)正常植株和花被片自然变异植株的外观形态差异很大,该研究以二者为材料,利用常规PCR和高效热不对称PCR(Hi-Tail PCR)技术从其正常和变异植株的基因组中各分离得到1个B类基因。序列分析证明,二者隶属于B类MADS-box基因AP3家族的旁系同源基因AP3-3分枝,分别命名为NArAP3-3(正常植株)和VArAP3-3(变异植株)。NArAP3-3基因全长3 795bp,VArAP3-3基因全长3 898bp,二者均含有1个666bp的开放阅读框(ORF),可编码221个氨基酸,具有典型的植物MADS-box基因结构,其编码肽链包含了MADS区、K区、Ⅰ区和C区。对比NArAP3-3和VArAP3-3基因的全长序列,发现VArAP3-3基因比NArAP3-3多了1段49bp的插入,且在ORF序列与NArAP3-3基因相比有4个碱基突变。对二者的全长序列、所编码的221个氨基酸及插入序列的生物信息学分析显示,二者在基因启动子、蛋白质基本性质、结构功能域、二级三级预测结构等方面均有差异,推测这些差异可能是花被片变异产生的原因之一。该研究结果为进一步探索其变异机制奠定了基础。     

Plasma concentrations of transforming growth factor beta 1 in non-progressive HIV-1 infection correlates with markers of disease progression

The human immunodeficiency virus (HIV) infection shows variable rate of disease progression. The underlying biological and molecular mechanisms involved in determining progression of HIV infection are not fully understood. The aims of this study were to determine plasma concentrations of active TGF β 1, Th1 and Th2 cytokines in patients with non-progressive and those with progressive HIV-1 infection, as well as to determine if there is an association of these cytokines to disease progression. In a cross-sectional study of 61 HIV-1 infected individuals categorized according to disease progression as having non-progressive HIV-1 infection (n = 14) and progressive infection (n = 47), plasma levels of active TGF β 1, INF-γ, TNF-α, IL-10, IL-1β, IL-12p70 and IL-13 were compared with HIV uninfected healthy controls (n = 12). Plasma concentration of these cytokines was measured using a highly sensitive luminex200 XMAP assay. Pearson correlation test was used to assess the correlation of cytokines with CD4+ and CD8+ T cells, CD4:CD8 ratio and plasma HIV-1 RNA in the different study groups. Plasma concentrations of TGF β 1 and IL-10 were significantly decreased while IL-1β, IL-12p70 and TNF-α were increased in patients with non-progressive HIV-1 infection compared to patients with progressive infection. Plasma levels of TGF β 1 and IL-10 showed an inverse correlation with CD8+ T cell counts and CD4:CD8 ratios in patients with non-progressive HIV-1 infection, while plasma HIV-1 RNA positively correlated with CD4+ T cell counts. Plasma levels of TNF-α, IL-1β, IL-12p70 and IL-13 positively correlated with CD4+ T cell counts and inversely correlated with plasma HIV-1 RNA, CD8+ T cell count and CD4:CD8 ratio in patients with non-progressive infection. The correlation of cytokines to the state of T-lymphocyte and plasma HIV-1 RNA found in this study may provide insight into the role of cytokines in both progressive and non-progressive HIV-1 infection. Additionally, these findings may have implications for systemic cytokine-based therapies in HIV-1 infection.     

鼻中隔成形术中鼻腔填塞法及鼻中隔“8”字缝合法的疗效比较研究

目的:探讨鼻中隔成形术中鼻腔填塞法及鼻中隔"8"字缝合法的疗效。方法:选取2017年7月~2018年7月期间我院收治的拟行鼻中隔成形术的鼻中隔偏曲患者60例。采用乱数表法将患者分为对照组、实验组两组,各30例患者,两组患者均行鼻内镜下鼻中隔成形术,术后对照组给予鼻腔填塞法处理,实验组给予鼻中隔"8"字缝合法处理。比较两组术后临床疗效、鼻粘膜水肿情况,观察两组不同时间点视觉疼痛模拟评分(VAS)情况,记录两组患者术后鼻中隔并发症发生情况。结果:实验组术后7d临床总有效率为96.67%(29/30),显著高于对照组患者的80.00%(24/30),差异有统计学意义(P0.05)。与术前相比,两组患者术后12hVAS评分升高,术后24h、术后48hVAS评分下降(P0.05);与术后12h相比,两组患者术后24h、术后48hVAS评分均下降,且术后48hVAS评分低于术后24h(P0.05);实验组术后12h、术后24h、术后48hVAS评分均低于对照组(P0.05)。实验组鼻粘膜水肿0级患者数量显著高于对照组,鼻粘膜水肿1级患者数量显著低于对照组(P0.05);两组鼻粘膜水肿2级患者数量比较差异无统计学意义(P0.05);两组患者鼻中隔并发症总发生率比较差异无统计学意义(P0.05)。结论:相较于鼻腔填塞法,鼻中隔成形术中采用鼻中隔"8"字缝合法疗效显著,可减轻患者疼痛,改善患者鼻粘膜水肿情况,且不会增加并发症发生率,具有一定的临床应用价值。     

CT灌注成像在兔早期肝硬化诊断中的应用价值研究

目的:探讨CT灌注成像在兔早期肝硬化诊断中的应用价值。方法:将55只新西兰大白兔随机分为2组,其中实验组45只,对照组10只。实验组给予皮下注射葡萄籽油稀释的50%CCL4,1次/4天,前4次剂量为1.0 m L/kg,第5次剂量为1.35 m L/kg,共注射20次。对照组采用同样方法只注射相同剂量的生理盐水。每注射4次后分别对实验组兔7只和正常对照组兔2只做螺旋CT灌注扫描,分析灌注参数,同时做相应的病理学观察,将二者进行比较及统计学分析。结果:注药4次末,兔血清ALT及AST明显高于注药前,注药8次末,兔血清ALT及AST最高,之后兔血清ALT及AST轻度减低,注药前后兔血清ALT及AST的变化有统计学意义(P0.05)。而血清(ALB)水平变化不明显,仅在注药16次末后,ALB水平稍减低,但差异无统计学意义。对照组肝脏灌注参数正常,实验组从注药4次开始,HAP呈上升趋势,但注药4次末及注药8次末,实验组及对照组之间差异无统计学意义(P0.1),注药12次末后二者之间差异有统计学意义(P0.05);而HPP、HBF及HBV呈下降趋势,MTT逐渐延长,与对照组的差异均有统计学意义(P0.05)。随兔血清ALT及AST的升高,HAP逐渐升高,MTT逐渐延长,而HPP、HBF及HBV逐渐减低。实验组肝小叶正常结构破坏,肝实质被纤维组织分割成大小不一、圆形或近圆形结节(假小叶),间隔较窄,炎症轻,结节边界尚整齐;汇管区内门脉小支扩张,壁增厚。对照组肝小叶结构规整,肝板排列有序,汇管区无扩大,其内个别炎细胞浸润,肝小叶内偶见点灶状坏死。结论:全肝CT灌注功能成像可为早期肝硬化的诊断提供影像学依据,将灌注征象与病理学变化结合有利于肝硬化的早期诊断和治疗。     

胃癌根治术后腹腔热灌注化疗对患者血清CEA、CA19-9水平及免疫功能的影响

目的:探讨胃癌根治术后腹腔灌注化疗对患者血清CEA(血清癌胚抗原,carcinoembryonic antigen)、CA19-9(糖链抗原19-9,carbohydrate antigen19-9)水平及免疫功能的影响。方法:回顾性2015年2月至2017年4月我院收治的胃癌患者临床资料,依据接受治疗方案不同分为全身静脉化疗组(对照组)和全身静脉化疗联合腹腔热灌注化疗组(观察组),每组各41例。检测和比较两组患者化疗前(治疗前)与化疗1个月后(治疗后)血清肿瘤标志物CEA、CA19-9与免疫功能指标水平的变化,治疗后毒副作用发生情况及治疗前后生活质量的改善情况。结果:治疗前,两组间血清CEA、CA19-9、CD3~+、CD4~+、CD8~+、CD4+/CD8~+水平比较差异均无统计学意义(P0.05);观察组治疗后血清CEA、CA19-9及CD8~+水平显著低于对照组,CD3~+、CD4~+、CD4~+/CD8~+水平显著高于对照组,差异有统计学意义(P0.05);两组骨髓抑制、恶心呕吐、腹痛腹泻及肠梗发生率比较差异均不显著无统计学意义(P=0.478,0.668,0.315,0.552);观察组生活质量改善总有效率为85.37%,显著高于对照组(70.73%,P=0.017)。结论:与单纯全身化疗相比,胃癌根治术后腹腔灌注化疗可更有效降低患者血清CEA、CA19-9水平,改善患者免疫功能,提高其生活质量,且安全性较高。     

藏山羊KLF8基因克隆及组织表达分析

本试验旨在获得藏山羊KLF8基因序列,并分析其生物学特征,同时阐明该基因在不同组织中的表达情况。利用RT-PCR技术克隆藏山羊KLF8基因序列,利用实时荧光定量PCR (quantitative real-time PCR,qPCR)检测其在藏山羊各个组织中的表达丰度。结果表明,获得藏山羊KLF8基因序列1 069 bp,其中包含CDS区1 008 bp,5'UTR序列28 bp和3'UTR序列33 bp,共编码335个氨基酸,为不稳定亲水碱性蛋白。KLF8基因在藏山羊的肺脏组织中表达水平最高,极显著高于其他组织(p<0.01)。本研究为进一步阐明KLF8基因在藏山羊中的生物学功能提供了依据。     

RNF8/RNF168信号通路在DNA损伤修复中的作用

细胞内DNA会受部分外界因素(如紫外辐射,电离辐射和化学毒素)和内部因素(如复制错误)的影响而发生损伤,包括DNA双链断裂、DNA错配和DNA交链等。DNA损伤发生后,损伤部位会被一些蛋白识别,进而招募一系列蛋白至损伤部位,形成一个修复系统。DNA双链断裂是最严重的一种DNA损伤,错误修复往往导致疾病的发生。DNA双链断裂(double strand break, DSB)后,细胞启动RNF8/RNF168信号通路进行修复。RNF8和RNF168是这条通路的枢纽蛋白;53BP和BRCA1是关键的效应蛋白,决定着DSB修复的方式;组蛋白泛素化、磷酸化和甲基化等翻译后修饰是这条通路顺利进行的基本条件;染色质重塑、泛素化酶/去泛素化酶平衡和蛋白稳定性是这条通路的主要调节方式。本综述对RNF8/RNF168信号通路进行了梳理总结,希望其能对相关研究者起到参考作用。