World Health Organization’s first-ever release of a fungal priority pathogens list:A reply action proposal for the prevention and treatment of fungal pathogens

Fungi are widely distributed in the environment,and some are beneficial to medicine,industry,agriculture,and food.For example,Penicillium chrysogenum,Acremonium chrysogenum,and Aspergillus terreus can synthesizeβ-lactams,which can be used as drugs[1];Aspergillus niger and Trichoderma reesei are well-known industrial enzyme producers[2].

Efficient genome editing in Claviceps purpurea using a CRISPR/Cas9 ribonucleoprotein method

Claviceps purpurea produces many pharmacologically important ergot alkaloids(EAS),which are widely used to treat migraine and hypertension and to aid childbirth.Although an EAS biosynthetic cluster of C.purpurea has been discovered more than 20 years ago,the complete biosynthetic pathway of EAS has not been fully characterized until now.The main obstacle to elucidating this pathway and strain modification is the lack of efficient genome-editing tools for C.purpurea.The conventional gene manipulation method for C.purpurea relies on homologous recombination(HR),although the efficiency of HR in C.purpurea is very low(~1-5%).Consequently,the disruption of target genes is laborious and time-consuming.Although CRISPR/Cas9 genome-editing methods based on in vivo Cas9 expression and gRNA transcription have been reported recently,their gene-disruption efficiency is still very low.Here,we developed an efficient genome-editing system in C.purpurea based on in vitro assembled CRISPR/Cas9 gRNA ribonucleoprotein complexes.As proof of principle,three target genes were efficiently knocked out using this CRISPR/Cas9 ribonucleoprotein complex-mediated HR system,with editing efficiencies ranging from 50%to 100%.Inactivation of the three genes,which are closely related to uridine biosynthesis(ura5),hypha morphology(rac),and EAS production(easA),resulted in a uridine auxotrophic mutant,a mutant with a drastically different phenotype in axenic culture,and a mutant that did not produce EAS,respectively.Our ribonucleoprotein-based genome-editing system has a great advantage over conventional and in vivo CRISPR/Cas9 methods for genome editing in C.purpurea,which will greatly facilitate elucidation of the EAS biosynthetic pathway and other future basic and applied research on C.purpurea.

妊娠合并子宫腺肌病患者胚胎着床部位与病灶位置的关系对妊娠结局的影响

目的探讨妊娠合并子宫腺肌病患者胚胎着床部位与病灶位置的关系对妊娠结局的影响。方法回顾性分析2018年1月至2020年12月因妊娠合并子宫腺肌病在浙江大学医学院附属妇产科医院住院治疗且可查到超声检查结果并能通过颈部透明层辨别胚胎或胎盘着床部位(≥孕11周)的95例患者的临床资料,复核超声检查结果,根据胚胎或胎盘着床部位与子宫腺肌病病灶位置的关系分为近距离组[指胚胎或胎盘着床部位在病灶上,或着床部位和病灶在子宫的同侧(即着床部位非常接近病灶),共59例]、远距离组(指胚胎或胎盘着床部位远离病灶,或着床部位和病灶在子宫的不同侧,共36例),进一步以妊娠28周为界将95例患者分为妊娠<28周(未能期待妊娠至28周即出现不良妊娠结局)和妊娠≥28周(妊娠至28周及以后),分析两组患者分别在这两个时间段中妊娠结局的差异。结果(1)95例妊娠合并子宫腺肌病患者的年龄为(34.8±3.5)岁。近距离组患者的年龄、孕前子宫大小以及初产妇、辅助生殖技术受孕、合并子宫内膜异位症、雌孕激素治疗史、病灶类型为弥漫性所占比例分别与远距离组比较,差异均无统计学意义(P均>0.05)。(2)95例患者中,妊娠<28周的患者12例(13%,12/95),其中近距离组11例(19%,11/59)、远距离组1例(3%,1/36),两组比较,差异有统计学意义(χ^(2)=5.100,P=0.027)。近距离组11例妊娠<28周出现不良妊娠结局的患者中,1例孕26周双胎妊娠先兆子宫破裂,5例妊娠中期胎死宫内,4例晚期难免流产,1例孕26周单胎活产;远距离组1例妊娠<28周出现不良妊娠结局的患者为晚期难免流产。(3)95例患者中,妊娠≥28周的患者83例(包括近距离组48例、远距离组35例),其最终妊娠结局均为活产。近距离组与远距离组相比,胎盘异常发生率(分别为60%、14%)、胎儿窘迫发生率(分别为27%、6%)、早产率(分别为67%、23%)、产时出血量[中位数分别为350 ml(范围:100~1500 ml)、300 ml(范围:100~800 ml)]均显著升高(P均<0.05),而孕周[中位数分别为37周(范围:30~41周)、38周(范围:28~41周)]、新生儿出生体重[中位数分别为2790 g(范围:1170~4040 g)、3010 g(范围:980~4320 g)]均显著降低(P均<0.05)。结论妊娠合并子宫腺肌病患者若胚胎着床部位位于病灶上或非常接近病灶,易发生不良妊娠结局,应严密监测、及早干预,以改善妊娠结局。