The head of sperm was imaged with domestic digital holographic microscopy (DHM), and then the quantitative three-dimensional size information of normal sperm and teratozoospermic sperm was compared and analyzed. DHM sperm imaging and repeated quantitative evaluation were used to determine the morphology of the sperm head in two patients with teratozoospermia and four volunteers with normal semen parameters. Sixty and 139 sperm of teratozoospermia patients and normal people were photographed by digital hologram, respectively. The differences in head height and width were compared and statistically analyzed. The sperm head height of the teratozoospermia group was 3.06 ± 1.66 μm, which was significantly lower than that of the normal sperm group (4.54 ± 1.60 μm, p < 0.01), but there was no significant difference in the head width between the two groups. Compared with the traditional two-dimensional optical microscope observation method, the DHM system can provide three-dimensional quantitative information for the sperm head and thus may help in the comprehensive clinical evaluation of the sperm head structure.

目的 探讨输卵管因素不育夫妇年龄和值或差值对体外受精-胚胎移植(IVF-ET)治疗结局的影响.方法 回顾性分析本中心2011年至2016年因输卵管因素接受IVF-ET治疗的1937对夫精夫妇和466对供精受精者的资料并随访至2019年9月.按男女年龄和值以及差值分组比较正常受精率、有效及优质胚胎数、临床妊娠率、流产率、累积活产率、婴儿出生体质量等.结果 夫妇一般会经过4年尝试自然受孕或常规治疗后选择IVF治疗;夫妇年龄和值分组与正常受精率无关,而与胚胎发育和移植后妊娠结局密切相关.当年龄和值≥70则有效胚胎数、优质胚胎数、临床妊娠率及活产率明显降低,和值≥80则产生明显的叠加累积效应(P＜0.01).夫妇年龄差值仅女方≥男方5岁会对妊娠率有统计学意义(P＜0.05).夫妇年龄和、差值与生化妊娠率、流产率、新生儿出生缺陷、低体质量儿之间差异无统计学意义(P＞0.05).结论 评估双方年龄和值比差值更有临床意义,双方会彼此弥补年龄对生育力的影响,双方都应尽量在40岁之前生育,减少年龄对生育和妊娠结果的影响.

OBJECTIVE:This was a prospective study to investigate whether progesterone affects sperm activity by regulating the cyclic AMP-protein kinase A (cAMP-PKA) signalling pathway via α/β hydrolase domain-containing protein 2 (ABHD2).METHODS:Spermatozoa were collected from healthy and infertile men (with oligoasthenospermia or abnormal acrosome; n = 30/group). The expression of and mutations in ABHD2 were detected by quantitative PCR, western blot, and gene sequencing. The expression of ABHD2 in the presence of progesterone was detected in all groups, and cAMP and PKA levels were detected by ELISA in fertile men after treatment with ABHD2 antibody and PKA inhibitor H-89, respectively.RESULTS:Expression of ABHD2 mRNA and protein were reduced in spermatozoa from infertile compared with fertile men. Four gene mutation sites were detected in spermatozoa from the infertile groups. Progesterone increased mRNA and protein levels of ABHD2 in healthy spermatozoa but not in spermatozoa from infertile men. The levels of cAMP and PKA were increased by progesterone in healthy spermatozoa, and the progesterone-increased cAMP and PKA were decreased by ABHD2 antibody and H-89, respectively.CONCLUSION:Progesterone regulates the ABHD2-mediated cAMP-PKA signalling pathway in healthy spermatozoa, which provides a new target for clinical diagnosis and treatment of infertility.

目的 研究ABHD2在不育症患者精子膜表面的表达差异,探究孕酮是否通过ABHD2调控cAMP-PKA进而影响精子的功能活性.方法 收集男性正常生育组精子、少弱精子组和顶体异常组精子样本(各30例),通过RT-PCR、蛋白Western-Blot印迹等方法分析转录水平及蛋白表达的差异,且采用体外添加孕酮、ABHD2抗体、PKA拟制剂H-89,选择观测正常生育组(对照组)与少弱精子组(实验组)男性精子ABHD2的蛋白表达水平,以及cAMP、PKA浓度等变化.结果 少弱精子组和顶体异常组中ABHD2 mRNA转录水平以及ABHD2蛋白表达水平显著减少,具有统计学意义(F:90.84,P＜0.01);在正常生育组中加入孕酮处理后,ABHD2蛋白水平表达明显增加;而少弱精子组中加入孕酮处理后,与未加孕酮相比,ABHD2蛋白水平表达增加不明显.进一步研究,在正常生育组中加入孕酮(P4)处理后,cAMP、PKA浓度极显著增加(P＜0.01),继续加入ABHD2抗体或PKA拟制剂H89后,cAMP、PKA浓度均显著减少(P＜0.01).结论 临床不育与正常生育男性精子之间ABHD2的转录和表达存在差异,孕酮通过ABHD2受体介导的cAMP-PKA信号通路调控精子受精功能.

睾酮制剂作为男性雄激素的替代治疗药物,在男性发育的不同年龄阶段具有不同的生理作用.因此,在临床发病不同年龄的男性中具有广泛应用,但效果不一.本文就睾酮替代治疗的临床应用(包括适应证、禁忌证、治疗益处与风险、临床制剂选择和随访)进行了综述,并结合临床最新的研究报道,为临床睾酮替代治疗研究及用药选择提供思路.

Background: The giant panda has an interesting bamboo diet unlike the other species in the order of Carnivora. The umami taste receptor gene T1R1 has been identified as a pseudogene during its genome sequencing project and confirmed using a different giant panda sample. The estimated mutation time for this gene is about 4.2 Myr. Such mutation coincided with the giant panda's dietary change and also reinforced its herbivorous life style. However, as this gene is preserved in herbivores such as cow and horse, we need to look for other reasons behind the giant panda's diet switch.Methodology/Principal Findings: Since taste is part of the reward properties of food related to its energy and nutrition contents, we did a systematic analysis on those genes involved in the appetite-reward system for the giant panda. We extracted the giant panda sequence information for those genes and compared with the human sequence first and then with seven other species including chimpanzee, mouse, rat, dog, cat, horse, and cow. Orthologs in panda were further analyzed based on the coding region, Kozak consensus sequence, and potential microRNA binding of those genes.Conclusions/Significance: Our results revealed an interesting dopamine metabolic involvement in the panda's food choice. This finding suggests a new direction for molecular evolution studies behind the panda's dietary switch.

The adaptive evolution of the rbcL gene in 14 species of Ephedra and 9 other gymnosperms was tested by the PAML program. Three large-scale selections were found in the catalytic region of the RbcL protein by branch model. Species in Ephedra with positive selection at the 365th codon site in rbcL gene were detected by branch-site model. 3D structure modeling showed that the 365th codon could change the spatial structure of the β-sheet (βG, βH, or βE) of the RbcL protein. These changes may be associated with the adaptation of the protein in Ephedra to the environment.

Incarvillea younghusbandii Sprague (Bignoniaceae) is a perennial herbaceous plant endemic to Qinghai-Tibetan Plateau. As a species of medical and horticulture importance, L younghusbandii is threatened by over exploitation and habitat fragmentation. In this study, we analyze the genetic diversity and population structure of I. younghusbandii using amplified fragment length polymorphism (AFLP) markers. Our data reveal very low levels of genetic diversity in seven natural populations across Tibet. Specifically, at population level, the average Nei's genetic diversity index (H-E) and Shannon's diversity index (1) were 0.063 and 0.096, respectively. In contrast, high genetic differentiation among populations (Gst = 0.6238, Phi(ST) = 0.614) is detected. The results of Neighbor-joining cluster, PCO, and STRUCTURE assignment reveal consistent pattern, suggesting seven well-defined genetic groups that are concordant with their geographical origins. The possible mechanisms and implications of these findings for conservation are discussed. (C) 2009 Elsevier Ltd. All rights reserved.

Araucariaceae is one of the most primitive families of the living conifers, and its phylogenetic relationships and divergence times are critically important issues. The DNA sequences of 8 genes, i.e., nuclear ribosomal 18S and 26S rRNA, chloroplast 16S rRNA, rbcL, matK and rps4, and mitochondrial coxI and atp1, obtained from this study and GenBank were used for constructing the molecular phylogenetic trees of Araucariaceae, indicating that the phylogenetic relationships among the three genera of this family should be ((Wollemia, Agathis), Araucaria). On the basis of the fossil calibrations of Wollemia and the two tribes Araucaria and Eutacta of the genus Araucaria, the divergence time of Araucariaceae was estimated to be (308 ± 53) million years ago, that is, the origin of the family was in the Late Carboniferous rather than Triassic as a traditional view. With the same gene combination, the divergence times of the genera Araucaria and Agathis were (246 ± 47) and (61 ± 15) Ma, respectively. Statistical analyses on the phylogenetic trees generated by using different genes and comparisons of the divergence times estimated by using those genes suggested that the chloroplast matK and rps4 genes are most suitable for investigating the phylogenetic relationships and divergence times of the family Araucariaceae.

南洋杉科是最原始的松杉纲类群之一,其系统发育关系与起源(分支)时间研究具有极为重要的意义.利用自行测定和GenBank中获得的核核糖体18S和26S,叶绿体16S,rbcL,matK和rps4以及线粒体cox 1和atp1等8个基因的DNA序列,构建了南洋杉科的分子系统树,确定该科属间系统发育关系应为((瓦勒迈杉属,贝壳杉属),南洋杉属).以瓦勒迈杉属和南洋杉属Araucaria族和Eutacta族的化石记录作为时间标定,估计南洋杉科的起源在(308±53)百万年前,即该科在石炭纪晚期起源,而非以往认为的三叠纪.用相同基因组合还估计出南洋杉属起源于(246±47)百万年前,贝壳杉属起源于(61±15)百万年前.对不同基因所构建的系统树进行统计分析并对它们所估计的起源时间进行比较表明,叶绿体matK和rps4基因较为适合南洋杉科的系统发育与起源时间估计研究.

Homogentisate Phytyltransferase(HPT)is a key enzyme catalyzing condensation of HGA and PDP in the first step of tocopherol biosynthesis.The HPT gene has been cloned from several photosynthetic organisms.Overexpression of HPT in transgenic plants increases tocopherols significantly in leaves and seeds.A mutation of the Arabidopsis HPT gene results in tocopherol deficiency in the plants.In order to understand the expression pattern of HPT,a 946 bp DNA upstream fragment of the Arabidopsis HPT gene was isolated and transcriptionally fused with the β-glucuronidase(GUS)reporter gene,and the promoter-GUS cassette was introduced into Arabidopsis thaliana by Agrobacterium-mediated transformation.Analysis of transgenic plants by histochemical staining showed that the GUS gene was expressed predominantly in the roots,and also to a lesser degree,in adult leaves,filament,and pistil,but not in stems,root tips,anthers,seeds,and siliqua.The results suggest that the HPT gene is predominantly expressed in Arabidopsis roots.

An Arabidopsis mutant,alr1, was isolated and characterized. The alr1 mutant exhibited downward curling and asymmetric rosette leaves. Moreover,it showed an enhanced apical dominance in shoots, agravitropism in roots, shortened hypocotyl and extended life span. It also exhibited a significant higher SOD activity in the leaves of mutant plants. Genetic analysis revealed that the mutant was due to a single recessive mutation,which was mapped to a region about 7.89 cM from nag111 on Chromosome 1 using SSLPs markers.

In an attempt to screen stay-green mutants in Arabidopsis in the absence of light, a mutant displaying the characteristics of brassinosteroids mutants was isolated. This mutant, dst6, carries a single recessive mutation. Using molecular genetic markers, the mutation was mapped to the bottom of chromosome II , which is around the region of DET2 ,one of the genes known to cause brassinosteroids mutation. The analysis of cDNA and genomic sequence of DET2 in dst6 revealed a frame-shifting insertion. These results suggest that dst6 is a putative new allele of det2.

Genetic analyses of lifespan in model animals have revealed that extended lifespans are closely associated to increased resistance to oxidative stress. In the model plant Arabidopsis, late-flowering mutants are also found to be more tolerant to oxidative stress. However, Arabidopsis mutants with extended lifespans are poorly studied so far. In this study, a screening system for mutants with extended lifespans in Arabidopsis was established using paraquat, a potent herbicide that exacerbates O2-. radical production. The relationship between lifespan and resistance to oxidative stress was examined with one of the mutants, SFNA-9-4. Compared to that of wild type, the lifespan of SFNA-9-4 is significantly extended, and its resistance to oxidative stress is also significantly elevated. These results suggest that, as in Drosophila, paraquat can also be used to screen for mutants with extended lifespans in Arabidopsis.