Genetic analysis of Pycr1 and Pycr2 in mice

The final step in proline biosynthesis is catalyzed by three pyrroline-5-carboxylate reductases, PYCR1, PYCR2, and PYCR3, which convert pyrroline-5-carboxylate (P5C) to proline. Mutations in human PYCR1 and ALDH18A1 (P5C Synthetase) cause Cutis Laxa (CL), whereas mutations in PYCR2 cause hypomyelinating leukodystrophy 10 (HLD10). Here, we investigated the genetics of Pycr1 and Pycr2 in mice. A null allele of Pycr1 did not show integument or CL-related phenotypes. We also studied a novel chemically-induced mutation in Pycr2. Mice with recessive loss-of-function mutations in Pycr2 showed phenotypes consistent with neurological and neuromuscular disorders, including weight loss, kyphosis, and hind-limb clasping. The peripheral nervous system was largely unaffected, with only mild axonal atrophy in peripheral nerves. A severe loss of subcutaneous fat in Pycr2 mutant mice is reminiscent of a CL-like phenotype, but primary features such as elastin abnormalities were not observed. Aged Pycr2 mutant mice had reduced white blood cell counts and altered lipid metabolism, suggesting a generalized metabolic disorder. PYCR1 and -2 have similar enzymatic and cellular activities, and consistent with previous studies, both were localized in the mitochondria in fibroblasts. Both PYCR1 and -2 were able to complement the loss of Pro3, the yeast enzyme that converts P5C to proline, confirming their activity as P5C reductases. In mice, Pycr1; Pycr2 double mutants were sub-viable and unhealthy compared to either single mutant, indicating the genes are largely functionally redundant. Proline levels were not reduced, and precursors were not increased in serum from Pycr2 mutant mice or in lysates from skin fibroblast cultures, but placing Pycr2 mutant mice on a proline-free diet worsened the phenotype. Thus, Pycr1 and -2 have redundant functions in proline biosynthesis, and their loss makes proline a semi-essential amino acid. These findings have implications for understanding the genetics of CL and HLD10, and for modeling these disorders in mice.     

Improvement of Opal Multiplex Immunofluorescence Workflow for Human Tissue Sections

The Opal multiplex technique is an established methodology for the detection of multiple biomarkers in one section. The protocol encompasses iterative single stainings and heating-mediated removal of the primary and secondary antibodies after each staining round, leaving untouched the Opal fluorophores which are deposited onto the antigen of interest. According to our experience, repetitive heating of skin sections often results in tissue damage, indicating an urgent need for milder alternatives to strip immunoglobulins. In this study, we demonstrate that considerable heating-related damage was found not only in skin but also in tissues of different origin, mostly characterized by low cell density. Importantly, the morphology remained fully intact when sections were repetitively exposed to β-mercaptoethanol-containing stripping buffer instead of multiple heating cycles. However, target epitopes appeared sensitive at a differential degree to multiple treatments with stripping buffer, as shown by loss in staining intensity, but in all cases, the staining intensity could be restored by increment of the primary antibody concentrations. Application of β-mercaptoethanol-containing stripping buffer instead of heating for antibody removal markedly improved the quality of the Opal multiplex technique, as a substantial higher number of differently colored cells could be visualized within a well-conserved morphological context:     

无创性皮肤屏障功能检测在朗格汉斯细胞组织细胞增生症中的应用

摘要 目的：探讨无创性皮肤屏障功能检测在朗格汉斯细胞组织细胞增生症(Langerhans cell histiocytosis,LCH)中的应用价值。方法：研究时间为2017年1月到2020年12月,选择在本院诊治的朗格汉斯细胞组织细胞增多症患者72例作为LCH组,同期选择健康体检者72例作为对照组。采用无创性皮肤屏障功能检测皮肤水分、经皮水分丢失(Transdermal water loss,TEWL)、油脂水平,同时检测所有入选者的免疫功能、皮肤菌群并进行相关性分析。结果：LCH组的皮肤水分低于对照组(P<0.05),TEWL、油脂水平高于对照组(P<0.05)。LCH组的乳酸杆菌(La)阳性率低于对照组(P<0.05),表皮葡萄球菌(Se)、痤疮丙酸杆菌(Pa)、金黄色葡萄球菌(Sa)阳性率高于对照组(P<0.05)。LCH组的CD163、ki-67表达阳性率分别为77.8 %、52.8 %,高于对照组的19.4 %和6.9 %(P<0.05)。在LCH组中,Pearson相关性分析显示皮肤水分与乳酸杆菌呈现正相关性(P<0.05),TEWL、油脂与表皮葡萄球菌、痤疮丙酸杆菌、金黄色葡萄球菌、CD163、ki-67呈现正相关性(P<0.05)。结论：无创性皮肤屏障功能检测在朗格汉斯细胞组织细胞增生症中的应用可反映患者的皮肤水分与油脂状况,也可间接反映患者的皮肤微生态与免疫功能状况。     

股前外侧穿支皮瓣与胸腹带蒂皮瓣对手外伤组织缺损修复的应用效果及对创面愈合程度的影响

摘要 目的：探讨股前外侧穿支皮瓣与胸腹带蒂皮瓣对手外伤组织缺损修复的应用效果及对创面愈合程度的影响。方法：选取我院2018年12月到2020年12月共收治的119例手外伤组织缺损患者作为研究对象,随机分为2组,分别为对照组（n=59,应用胸腹带蒂皮瓣修复术）和观察组（n=60,应用股前外侧穿支皮瓣修复术）。对比两组患者治疗优良率,对比两组患者治疗前后手部创面面积、创面愈合程度以及组织愈合时间,对比两组患者治疗后的Jamar握力、TAM和DASH评分情况,对比两组患者的皮瓣成活率、皮瓣危象率和血管吻合时间。结果：通过对比两组患者治疗优良率发现,观察组患者优的人数为21例、良为35例,优良率为93.33%,对照组患者优的人数为16例,良为30例,优良率为77.97%,观察组高于对照组（P＜0.05）;治疗后,与对照组相比,观察组患者的手部创面面积、组织愈合时间和DASH评分显著减少,创面愈合程度以及TAM与Jamar握力显著增加（P＜0.05）;通过对比两组患者的皮瓣成活率、术后皮瓣危象率以及血管吻合时间发现,两组患者的术后皮瓣危象率、血管吻合时间对比无明显差异（P＞0.05）,两组患者的术后皮瓣成活率对比差异显著,观察组明显高于对照组（P＜0.05）。结论：对手外伤组织缺损患者应用股前外侧穿支皮瓣与胸腹带蒂皮瓣修复术均具有明显的修复效果,但是应用股前外侧穿支皮瓣能够提升治疗效果,提升患者创面愈合程度减少愈合时间,提升患者手部运动情况,提升术后皮瓣成活率,值得临床应用推广。     

甘草叶片形态结构和光合作用对干旱胁迫的响应

叶片结构在植物防御生物和非生物胁迫方面起着重要的作用,可通过合成、储存和分泌次生代谢产物提高植物抗性。以甘草幼苗为试材,采用盆栽控水自然干旱法,探讨叶片光合作用、气孔微形态和腺体形态对干旱胁迫的响应。结果表明:①随着干旱胁迫程度的加剧,叶片净光合速率(Pn)、气孔导度(Gs)和蒸腾速率(Tr)均呈先升高后降低的趋势;其中胞间CO2浓度(Ci)在重度干旱胁迫(severe stress,SS)时迅速增高。②随着干旱胁迫程度的加剧,叶片总气孔密度和气孔开张比呈先增大后减小的趋势;而气孔开张宽度呈逐渐减小的趋势。③随着干旱胁迫程度的加剧,叶片上表皮和下表皮腺体密度总数整体上呈增大的趋势,腺体颜色随着干旱胁迫程度的加剧逐渐加深,形状出现不规则褶皱和内陷。总之,甘草叶片表面的腺体特征参与抗旱逆境调节,从而避免干旱胁迫对甘草植株的伤害;在SS下,胁迫程度加速了气孔细胞的程序性死亡(PCD),甘草幼苗失去抗旱能力。     

宏基因组学技术在痤疮研究中的应用进展

高通量测序技术的发展提高了人们对微生物组的认识。宏基因组学技术因其全面和深入的分析功能被广泛应用于各种环境微生物组的研究中,尤其在阐明各种疾病与人体微生物组的关系中,宏基因组学技术具有重要作用。痤疮作为一种常见的皮肤疾病,严重影响人们皮肤美观度和心理健康。利用宏基因组学技术挖掘皮肤微生物与痤疮的关系,将有助于痤疮发病机理的研究和临床治疗方法的改进。通过介绍宏基因组学技术的发展背景、概述及其应用研究进展,探讨皮肤微生物与痤疮的关系,综述宏基因组学技术在痤疮研究中的应用现状,并总结目前宏基因组学技术在皮肤疾病研究中存在的问题,旨在为痤疮的宏基因组研究提供参考。     

Adaptive morphological shifts to novel habitats in marine sculpin fishes

Sculpin fishes of the North American Pacific Coast provide an ideal opportunity to examine whether adaptive morphological character shifts have facilitated occupation of novel habitat types because of their well‐described phylogeny and ecology. In this group, the basal‐rooted species primarily occupy the subtidal habitat, whereas the species in the most distal clades are found in the intertidal. We tested multiple evolutionary models to determine whether changes in body size and changes in number of scales are adaptive for habitat use in sculpins. Based on a statistically robust, highly resolved molecular phylogeny of 26 species of sculpins, in combination with morphometric and habitat affinity data, our analyses show that an adaptive model based on habitat use best explains changes in body size and number of scales. The habitat model was statistically supported over models of neutral evolution, stabilizing selection across all habitats, and three clade‐based models. We suggest that loss of scales and reduction of body size in the intertidal may facilitate cutaneous breathing in air when tidepools become hypoxic during low tides. This study demonstrates how the combined use of phylogenetic, ecological and statistical approaches helps to identify traits that are likely adaptive to novel habitats.