Universal two-dimensional labelled probe-mediated melting curve analysis based on multiplex PCR for rapid typing of Plasmodium in a single closed tube

Currently, malaria is still one of the major public health problems commonly caused by the four Plasmodium species. The similar symptoms of malaria and the COVID-19 epidemic of fever or fatigue lead to frequent misdiagnosis. The disadvantages of existing detection methods, such as time-consuming, costly, complicated operation, need for experienced technicians, and indistinguishable typing, lead to difficulties in meeting the clinical requirements of rapid, easy, and accurate typing of common Plasmodium species. In this study, we developed and optimized a universal two-dimensional labelled probe-mediated melting curve analysis (UP-MCA) assay based on multiplex and asymmetric PCR for rapid and accurate typing of five Plasmodium species, including novel human Plasmodium, Plasmodium knowlesi (Pk), in a single closed tube following genome extraction. The assay showed a limit of detection (LOD) of 10 copies per reaction and could accurately distinguish Plasmodium species from intra-plasmodium and other pathogens. Additionally, we proposed and validated different methods of fluorescence quenching and tag design for probes that are suitable for UP-MCA assays. Moreover, the clinical performance of the Plasmodium UP-MCA assay using a base-quenched universal probe was evaluated using 226 samples and showed a sensitivity of 100% (164/164) and specificity of 100% (62/62) at a 99% confidence interval, with the microscopy method as the gold standard. In summary, the UP-MCA assay showed excellent sensitivity, specificity, and accuracy for genotyping Plasmodium species spp. Additionally, it facilitates convenient and rapid Plasmodium detection in routine clinical practice and has great potential for clinical translation.     

平菇丰收一号高产特性的研究

通过野生种分离选用的平菇丰收一号,染色体数为９（ｎ＝９）,菌丝生长速度比一般品种快２—３４ｃｍ／ｈ,平均投料０．５ｋｇ（块）产鲜菇０．６～２．５ｋｇ,个别单株重１０～２０ｋｇ左右,生物效率１００～１５０％,高者达２００％以上,是一种高产优质、适应性强的新菌种。     

朱砂叶螨羧酸脂酶最优测试条件的选择

应用二次回归通用旋转组合设计,对朱砂叶螨离体羧酸酯酶测试过程中所需缓冲液pH值、恒温时间、反应温度及底物浓度,设立4因子5水平试验,在考虑4个因子主效应和互作效应的情况下,筛选测试羧酸酯酶的最优条件组合。     

Evidence for FUS6 as a component of the nuclear-localized COP9 complex in Arabidopsis.

The pleiotropic CONSTITUTIVE PHOTOMORPHOGENIC (COP), DEETIOLATED (DET), and FUSCA (FUS) loci are essential regulatory genes involved in the light control of seedling developmental patterns in Arabidopsis. Although COP1, DET1, COP9, and FUS6 (also called COP11) have been cloned, their biochemical activities and interactions remain elusive. We have recently suggested that multiple pleiotropic COP, DET, and FUS genes may encode subunits of a large regulatory complex. In this study, we generated specific antibodies against Arabidopsis FUS6 and show that accumulation of both COP9 and FUS6 is coordinated in the pleiotropic cop, det, and fus mutant backgrounds and in wild-type plants throughout development. Both COP9 and FUS6 cofractionated into identical high molecular mass fractions in an analytical gel filtration assay, and neither was found in its monomeric form. Moreover, antibodies raised against either COP9 or FUS6 selectively coimmunoprecipitated both proteins. We have also developed an Arabidopsis protoplast immunolocalization assay and demonstrated that the COP9 complex is localized in the nucleus and that its nuclear localization is not affected by light conditions or tissue types. The integrated genetic and biochemical results strongly support the conclusion that both COP9 and FUS6 are components of the nuclear-localized COP9 complex. Therefore, we have provided the strongest evidence for the conclusion that at least some of the pleiotropic COP, DET, and FUS loci act in the same signaling pathway.     

Purification, properties, and immunological characterization of GalT-3 (UDP-galactose: GM2 ganglioside, β1-3 galactosyltransferase) from embryonic chicken brain

A 1-3 galactosyltransferase (GalT-3; UDP-Gal; GM2 1-3galactosyltransferase) was purified over 5100-fold from 19-day-old embryonic chicken brain homogenate employing detergent solubilization, -lactalbumin Sepharose, Q-Sepharose, UDP-hexanolamine Sepharose, and GalNAc1-4Gal-Synsorb column chromatography. The purified enzyme was resolved into two bands on reducing gels with apparent molecular weights of 62 kDa and 65 kDa, respectively. GalT-3 activity was also localized in the same regions by activity gel analysis and sucrose-density gradient centrifugation of a detergent-solubilized extract of 19-day-old embryonic chicken brain. Purified GalT-3 exhibited apparentK _mS of 33 µm, 22 µm and 14.4mM with respect to the substrates GM2, UDP-galactose, and MnCl₂, respectively. Substrate specificity studies with the purified enzyme and a variety of glycosphingolipids, glycoproteins, and synthetic substrates revealed that the enzyme was highly specific only for the glycosphingolipid acceptors, GM2 and GgOse3Cer (asialo-GM2). Ovine-asialo-agalacto submaxillary mucin inhibited the transfer of galactose to GM2 but did not act as an acceptor in the range of concentrations tested. Polyclonal antibodies raised against purified GalT-3 inhibited GalT-3 activityin vitro and Western-immunoblot analysis of purified GalT-3 showed immunopositive bands at 62 and 65 kDa.Abbreviations CNS central nervous system - GM1 monosialotetraosylganglioside, Gal1-3GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer - GM2 monosialotriaosylganglioside, GalNAc1-4(NeuAc2-3)Gal1-4Glc1-1Cer - DSS detergent solubilized supernatant - ECB embryonic chicken brain - TBS Tris-buffered saline     

粘虫颗粒体病毒的增效因子提高杆状病毒的感染

一株美洲粘虫GV(增效品系)在东方粘虫幼虫上进行增殖,所获粘虫GV(PuGv-Ps>对东方粘虫NPV(PsNPV)、棉铃虫NPV(HaNPV)和黄地老虎NPV(AsNPV)进行增效试验。 实验结果证明PuGV-Ps对三种NPV都有明显的增效作用, 其中以对PsNPV为最强,增效率达55%一85%;对HaNPV为30%-80%;而对AsNPV只有15%-35%。用凝胶过滤技术从PuGV-Ps中分离增效因子PusF-Ps,其对PsNPV和A,NPV的增效作用亦同样明显,250μg的PuSF-Ps可以提高PsNPV的感染能力达80%。经ScPhadcf-150柱(1.6X 90)和使用四种标准蛋白(细胞色素c、胃蛋白酶、牛血清清蛋白和乳酸脱氢酶)测得PuSF-Ps的分子量为160 000左右。     

西藏林芝真蚋亚属三新种（双翅目：蚋科）

本文记述西藏林芝真蚋亚属Ｅｕｓｉｍｕｌｉｕｍ三种：凸端真蚋Ｓｉｍｕｌｉｕｍ（Ｅｕｓｉｍｕｌｉｕｍ）ｃｏｎｃａｖｕｓｔｙｌｕｍｓｐ．ｎｏｖ．、林芝真蚋Ｓｉｍｕｌｉｕｍ（Ｅｕｓｉｍｕｌｉｕｍ）ｌｉｎｇｚｉｅｎｓｅｓｐ．ｎｏｖ．、裂缘真蚋Ｓｉｍｕｌｉｕｍ（Ｅｕｓｉｍｕｌｉｕｍ）ｓｃｈｉｚｏｌｏｍｕｎｓｐ．ｎｏｖ 。     

买麻藤茎的结构及其异常次生生长

买麻藤(Gnetum montanum Mgr.)茎的各类组织的排列与被子植物的茎非常相似,其次生生长也属同一类型。其中最明显的相似之处,是木质部中的导管和韧皮部中的筛分子与“伴胞”。藤本植物买麻藤,正常的次生生长开始后,由每一维管束外侧的薄壁组织经脱分化产生新的形成层,以后逐渐形成一圈维管束。这与前人所描述的新形成层来自韧皮薄壁组织是不同的。异常的维管束与正常的一样,由木质部和韧皮部组成,并被髓射线分隔呈楔形。当第一轮维管束停止生长以后,在其外围以同样方式形成新的一轮,以后并可连续形成多轮。     

ph1b基因在Aegilops有益基因直接遗传转移中利用的可能性

第一次用中国春和中国春ph1b突变体对(中国春phlb突变体×Ae.uariabilis)F_1和(中国春ph1b突变体×Ae.turcomenica)F_1回交获得了成功,并通过连续回交,把Ae.turcomenica的抗白粉基因转移到了普通小麦中。证实了利用ph1b基因从山羊草属的一些种“直接遗传转移”有益基因到普通小麦中的可能性。     

急进高海拔时健康人夜间睡眠、呼吸状态和血氧饱和度的变化

在海拔2300m选择健康成年男性5人,急进抵海拔4660m,用多导监测仪分别在两地连续7h监测夜间睡眠、呼吸状态和血氧饱和度变化,进行自身对比。结果发现:(1)急进高海拔后,总睡眠时间、有效睡眠指数、Ⅲ～Ⅳ期深睡眠均较中度高原减少(p<0.01);总觉醒时间、Ⅰ～Ⅱ期浅睡眠高海拔较中度高原增多(p<0.05):(2)急进高海拔后,有3名健康人出现周期性呼吸,其中1名健康者出现周期性呼吸119次,伴有中枢性睡眠呼吸暂停,最低Sao_2为78％;(3)同海拔高度夜间睡眠时与清醒时Sao_2相比较,中度高原下降4.2％,高海拔下降11.2％(p<0.01);高海拔与中度高原夜间清醒时Sao_2相比较下降7.4％,睡眠时下降14.4％(p<0.001)。结果提示:(1)睡眠加重了高原人原有的低氧血症;(2)低氧血症导致睡眠结构的紊乱和睡眠质量的降低;(3)睡眠中出现的周期性呼吸,应视为机体的一种自我保护机制;(4)频发的周期性呼吸或睡眠呼吸暂停将影响大脑机能。