Clinical,Virological and Immunological Features from Patients Infected with Re-Emergent Avian-Origin Human H7N9 Influenza Disease of Varying Severity in Guangdong Province

BackgroundThe second wave of avian influenza H7N9 virus outbreak in humans spread to the Guangdong province of China by August of 2013 and this virus is now endemic in poultry in this region.MethodsFive patients with H7N9 virus infection admitted to our hospital during August 2013 to February 2014 were intensively investigated. Viral load in the respiratory tract was determined by quantitative polymerase chain reaction (Q-PCR) and cytokine levels were measured by bead-based flow cytometery.ResultsFour patients survived and one died. Viral load in different clinical specimens was correlated with cytokine levels in plasma and broncho-alveolar fluid (BALF), therapeutic modalities used and clinical outcome. Intravenous zanamivir appeared to be better than peramivir as salvage therapy in patients who failed to respond to oseltamivir. Higher and more prolonged viral load was found in the sputum or endotracheal aspirates compared to throat swabs. Upregulation of proinflammatory cytokines IP-10, MCP-1, MIG, MIP-1α/β, IL-1β and IL-8 was found in the plasma and BALF samples. The levels of cytokines in the plasma and viral load were correlated with disease severity. Reactivation of herpes simplex virus type 1(HSV-1) was found in three out of five patients (60%).ConclusionExpectorated sputum or endotracheal aspirate specimens are preferable to throat swabs for detecting and monitoring H7N9 virus. Severity of the disease was correlated to the viral load in the respiratory tract as well as the extents of cytokinemia. Reactivation of HSV-1 may contribute to clinical outcome.     

VK_3诱导悬浮培养的胡萝卜细胞凋亡

VK_3是一种醌类物质,在体内通过氧化还原产生超氧化自由基。我们的实验发现,VK_3对悬浮培养的胡萝卜细胞及原生质体有致死作用。这种作用是浓度依赖性的。100—800μmol/L的VK_3能引起10%—33%的细胞死亡。当VK_3浓度达到1mmol/L时,死亡率达到100%。DNA电泳分析发现,600和800μmol/L的VK_3处理过的细胞和原生质体产生大小为180bp整数倍的梯状条带。而在更高或更低浓度的处理中则没有观察到这种条带。用TUNEL方法检测,在400—800μmol/L浓度的处理中观察到细胞核DNA的断裂。而在更高或更低浓度的处理中则没有观察到这种断裂。由此我们推测,适当浓度的VK_3能诱导胡萝卜细胞及原生质体凋亡。     

遮荫对‘保佳红’桃树叶片快速叶绿素荧光诱导动力学曲线的影响

以3年生桃品种‘保佳红’为试材,于不同生育时期采用人工遮阴的方法,在叶片生长的不同时期分别设置100%(CK)、80%、60%、40%和20% 自然光照的5个光照强度处理,利用快速叶绿素荧光诱导动力学曲线分析技术,研究了不同光强对桃树叶片快速叶绿素荧光诱导动力学曲线及其参数的影响。结果表明：（1）桃树各生长时期的叶片最大荧光值均随着光照强度的减弱而依次升高。（2）在PSⅡ能量分配比率方面,遮阴下的叶片提高了用于电子传递的量子比率(φ_Eo),降低了用于热耗散的的量子比率(φ_Do)。（3）在PSⅡ反应中心活性方面,遮阴使得单位反应中心吸收的光能(ABS/RC)、捕获的光能(TRo/RC)、用于传递电子的能量(ETo/RC)和用于热耗散的能量(DIo/RC)均下降。（4）各时期不同光强处理的最大光化学效率F_v/F_m均在中午时段出现下降,且光照强度越大,降幅越大,说明桃叶在中午会出现强光抑制。研究认为：在遮阴条件下,桃叶天线色素吸收和捕获的光能减少,PSⅡ反应中心活性降低,但其可以通过增加能量在电子传递方面的分配比率来提高对光能的利用。     

miR-29a attenuates cardiac hypertrophy through inhibition of PPARδ expression

Although cardiac hypertrophy is widely recognized as a risk factor that leads to cardiac dysfunction and, ultimately, heart failure, the complex mechanisms underlying cardiac hypertrophy remain incompletely characterized. The nuclear receptor peroxisome proliferator-activated receptor δ (PPARδ) is involved in the regulation of cardiac lipid metabolism. Here, we describe a novel PPARδ-dependent molecular cascade involving microRNA-29a (miR-29a) and atrial natriuretic factor (ANF), which is reactivated in cardiac hypertrophy. In addition, we identify a novel role of miR-29a, in which it has a cardioprotective function in isoproterenol hydrochloride-induced cardiac hypertrophy by targeting PPARδ and downregulating ANF. Finally, we provide evidence that miR-29a reduces the isoproterenol hydrochloride-induced cardiac hypertrophy response, thereby underlining the potential clinical relevance of miR-29a in which it may serve as a potent therapeutic target for heart hypertrophy treatment.     

Cellophane surface-induced gene,VdCSIN1, regulates hyphopodium formation and pathogenesis via cAMP-mediated signalling in Verticillium dahliae

The soil-borne vascular pathogen Verticillium dahliae infects many dicotyledonous plants to cause devastating wilt diseases. During colonization, V. dahliae spores develop hyphae surrounding the roots. Only a few hyphae that adhere tightly to the root surface form hyphopodia at the infection site, which further differentiate into penetration pegs to facilitate infection. The molecular mechanisms controlling hyphopodium formation in V. dahliae remain unclear. Here, we uncovered a cellophane surface-induced gene (VdCSIN1) as a regulator of V. dahliae hyphopodium formation and pathogenesis. Deletion of VdCSIN1 compromises hyphopodium formation, hyphal development and pathogenesis. Exogenous application of cyclic adenosine monophosphate (cAMP) degradation inhibitor or disruption of the cAMP phosphodiesterase gene (VdPDEH) partially restores hyphopodium formation in the VdΔcsin1 mutant. Moreover, deletion of VdPDEH partially restores the pathogenesis of the VdΔcsin1 mutant. These findings indicate that VdCSIN1 regulates hyphopodium formation via cAMP-mediated signalling to promote host colonization by V. dahliae.     

超声引导下前列腺穿刺联合外周血循环肿瘤细胞检测对前列腺癌预后分析

目的探讨超声引导下前列腺穿刺联合外周血循环肿瘤细胞(CTCs)检测对前列腺癌预后的预测效果。方法选取2011年1月至2017年12月期间于郑州大学第二附属医院收治的83例前列腺癌患者为研究对象,全部患者均根据超声引导下经直肠前列腺穿刺活检术确诊为前列腺癌,检测病理标本中CK34BE12、p63、α-甲酰基辅酶A消旋酶(AMACR)等免疫标志物的表达状况,并采用Cell Search细胞搜索系统检测外周血CTCs的数量,据此分为阳性组(≥5个/7.5 ml)和阴性组(<5个/7.5 ml)。分析穿刺组织中免疫标志物表达状况、外周血CTCs计数与患者临床病理特征、生存状况的相关性。各标志物的阳性例数、Gleason评分>7分的比例、TNM分期等定性资料的比较采用x^2检验或Fisher确切概率法,年龄、血常规、凝血功能、肝功能、PSA水平等定量资料的比较采用t检验。采用Kaplan-Meier法进行生存分析,采用多因素Cox比例风险回归模型分析患者预后的预测因素。结果(1)全部患者中外周血CTCs、穿刺组织中CK34BE12、p63、AMACR的阳性率分别为31.33、3.61、3.61、86.75。CTCs阳性组的AMACR阳性率为100.00,高于CTCs阴性组的80.70,差异有统计学意义(x^2=4.227,P<0.05)。(2)AMACR阳性组患者的血红蛋白(HB)低于AMACR阴性组[(123.66±13.33)g/L比(134.89±20.08)g/L,t=2.420,P=0.018],血小板(PLT)、血清谷丙转氨酶、D-二聚体(DD)、前列腺特异抗原(PSA)水平、Gleason评分>7分的比例均高于AMACR阴性组[(197.23±36.98)×10^9/L比(172.83±33.33)×10^9/L,t=2.062,P=0.042;(38.80±10.03)U/L比(31.46±7.83)U/L,t=2.317,P=0.023;(255.00±38.80)μg/L比(220.81±30.99)μg/L,t=2.785,P=0.007;(26.60±12.23)ng/ml比(17.90±8.88)ng/ml,t=2.263,P=0.026;45.83比9.09,x^2=3.916,P=0.048],差异有统计学意义(P<0.05)。CTCs阳性组患者的HB低于CTCs阴性组[(121.69±15.89)g/L比(132.73±18.85)g/L,t=2.767,P=0.007],血清碱性磷酸酶、DD、PSA水平、Gleason评分>7分、T3~T4期、M1期的比例均高于CTCs阴性组[(105.69±30.56)U/L比(88.89±35.58)U/L,t=2.205,P=0.030;(256.63±35.86)μg/L比(236.98±33.30)μg/L,t=2.368,P=0.020;(30.09±11.89)ng/ml比(23.33±10.99)ng/ml,t=2.533,P=0.013;57.69比33.33,x^2=4.381,P=0.036;30.77比8.77,x^2=4.981,P=0.026;50.00比17.54,x^2=9.390,P=0.002],差异有统计学意义(P<0.05)。(3)全部患者的中位生存时间为58.33个月,1、3、5年的生存率分别为88.95、51.81、30.12。AMACR阳性组、CTCs阳性组患者的中位生存时间为40.93、36.93个月,低于AMACR阴性组、CTCs阴性组的66.66、69.56个月,差异有统计学意义(P<0.05)。多因素Cox比例风险回归模型分析结果表明,Gleason评分>7分、M1期、AMACR阳性、CTCs阳性是患者死亡的独立危险因素(HR=1.883、3.666、2.009、2.923,P<0.05)。结论超声引导下前列腺穿刺联合外周血CTCs检测对前列腺癌患者的预后具有重要的预测价值,临床上可根据穿刺组织中AMACR表达水平和外周血CTCs计数进行预后的综合分析。     

A natural product enhances apoptosis via mitochondria/caspase-mediated pathway in HeLa cells

Cervical cancer is the fourth most lethal human malignancy and the leading cause of death among females around the world. Many antitumor agents have microbial origins. 5′-epi-SPA-6952A is a new 24-membered macrolide isolated from the cultured broth of Streptomyces diastatochromogenes. Therefore, we studied the activity and molecular mechanism of 5′-epi-SPA-6952A in human cervical carcinoma HeLa cell. The results showed that 5′-epi-SPA-6952A significantly inhibited cell proliferation and migration. In addition, 5′-epi-SPA-6952A obviously increased the production of intracellular reactive oxygen species and DNA damage in HeLa cells. Moreover, nuclear shrinkage of cells, decrease in mitochondrial membrane potential, and upregulation of Bax/Bcl-2 ratio resulted in the release of cytochrome c, and activation of caspase-9/3 was observed in HeLa cells treated with 5′-epi-SPA-6952A, which means it enhanced the intrinsic mitochondrial apoptosis. Besides, DNA-damage associated proteins poly (ADP-ribose) polymerase (PARP) and p53 were also studied, and the expressions of cleaved-PARP and p53 were drastically increased in HeLa cells treated with 5′-epi-SPA-6952A. Furthermore, we confirmed that 5′-epi-SPA-6952A affected the survival of HeLa cells by blocking cell cycle progression in the G1 phase. Taken together, the results shows that 5′-epi-SPA-6952A significantly inhibited HeLa cells proliferation via intrinsic mitochondrial apoptosis, cell cycle arrest, and blocking cell migration.