论前列腺为“精胞”

前列腺是男性重要的性腺器官,在中医古代医籍中并无前列腺一词的记载,现医家多将其归属于精室,然精室涵盖范围较广,主要指男性生殖系统。因此为明确前列腺的中医定位,基于中医关于胞的认识以及前列腺的现代研究,将其与女子胞(胞宫)相对比,提出可将前列腺的中医名称命名为精胞。

恙螨体内恙虫病立克次体经精胞传递的实验研究

本文报道地里纤恙螨雄虫人工接种恙虫病立克次体后与雌虫配对培养和传代，应用幼虫叮咬小鼠分离立克次体和PCR结合核酸杂交（核酸杂交检测PCR产物）检测子代体内立克次体的结果。幼虫叮咬小鼠分离立克农作检查第3子代幼虫体内立克次体阳性。PCR结合核酸杂交检测佛山市的病人恙虫病立克次休（未定株）接种雄虫和健康雌虫配对的第1、2、3子代成虫体内立克次体DNA阳性。Karp株接种雄虫的第1子代幼虫体内立克次体的幼虫叮咬小鼠法和PCR结合核酸杂交均呈阳性。

五种恙螨精胞(真螨目:恙螨科)(恙螨研究 ⅩⅨ)

一、绪 言 与氏阿康恙螨Acomatacarus yosanoi Fukuzumi et Obata,1953的精胞发现及其间接交配试验的结果,促使我们进一步考虑这一现象的普遍性问题。 1957年春季中国人民解放军军事医学科学院获得了大批恙螨幼虫,并在实验室中饲养到达成虫,为了帮助我们了解这一生活史特性,故在同年5月和8月之间把饲养所得的成虫供给我们观察了一个时期,这些成虫是地里恙螨Trombicula deliensis Walch,1923,

黄欲晓教授运用填精荣胞法分期治疗复发性流产经验

总结黄欲晓教授运用填精荣胞法分期治疗复发性流产(Recurrent spontaneous abortion,RSA)的临证经验。黄欲晓教授认为肾精亏虚是RSA的基本病机,虚久致瘀,病传肝脾,内、外、杂三因合而为病,终致精气血紊乱,胞宫不荣不通,屡孕屡堕。基于RSA发病及传变特点,黄欲晓教授认为其治疗关键在于将调治端口前移,未病先防;强调立足病机,以填精荣胞为治疗大法,补肾填精贯穿疾病治疗始终;在孕前调经以助胞宫藏泻,荣养胞宫,预培其损;在孕后积极补肾安胎,随症加减,去其所病。

左旋肉碱在经皮附睾穿刺取精-卵细胞胞质内单精子注射治疗中的应用

目的:探讨左旋肉碱在经皮附睾穿刺取精-卵细胞胞质内单精子注射(PESA-ICSI)治疗中的应用。方法:随机将2008年9月至2009年8月间本中心就治的79例梗阻性无精子症患者分成2组:未服药组(A组36例)和服药组(B组43例)。B组ICSI治疗取卵前3个月口服左旋肉碱(1g,2次/d)治疗,A组未作任何治疗作为对照,比较A、B两组间PESA-ICSI的获卵数、受精数、受精率、优质胚胎数和优质胚胎率。结果:A、B两组间受精数和受精率无显著性差异,但B组获得优质胚胎数及优质胚胎率明显高于A组(P<0.05)。结论:ICSI治疗前不育男性口服左旋肉碱3个月能显著提高PESA-ICSI的优质胚胎数及优质胚胎率,为将来获得良好的妊娠结局奠定基础。

补肾活血导法改善PER“胞宫摄精成孕”的网络调控机制研究

目的基于“精满则子宫易于摄精,血足则子宫易于容物”思想,通过实验研究结合网络分析、分子对接等方法,探讨补肾活血方治疗子宫内膜容受性不良(Poor endometrial receptivity,PER)的关键核心靶点及信号通路的作用机制。方法建立肾虚血瘀-PER病证结合大鼠模型,补肾活血导法干预10天后,通过扫描电镜观察大鼠的种植期胞饮突形态;利用TCMSP数据库结合Batman数据库筛选补肾活血方中有效组分,通过Genecards、OMIM数据库检索PER基因;筛选补肾活血方-PER共同作用靶点,导入String数据库及Cytoscape3.8.0软件进行可视化分析,构建PPI网络;应用Metascape数据库分别进行GO富集分析及KEGG富集分析。采用AutoDock对补肾活血方有效组分与核心靶标蛋白进行分子对接验证,通过Western Blotting、q-PCR和ELISA检测相关关键基因蛋白或mRNA的表达水平。结果补肾活血导法可提高PER大鼠着床率,促进胞饮突成熟。补肾活血方与PER共有130个交集靶点;String计算后应用Cytoscape中CytoNCA插件获得24个核心靶点,输入metascape平台获得相关66个生物学过程(BP)、38个分子功能(MF)、28个细胞组分(CC)和相关KEGG信号通路共113条。分子对接提示补肾活血方有效组分与关键蛋白具有良好的亲和力作用。实验研究显示补肾活血导法能够增强TNF、p-38、ESR1、PGR、IL6、1L1β等相关因子在子宫内膜组织上的表达。结论通过网络分析结合药理实验研究发现,补肾活血导法能够有效改善PER相关指标,并通过调控IL6、IL-1β、p-38蛋白和基因表达水平,改善子宫内膜容受性进而实现“胞宫摄精成孕”。

基于养精育胞论构建中医妇科肾虚类疾病治疗新体系——国医大师何成瑶学术思想与临床经验研究

国医大师何成瑶重视“养精”“育胞”在女性生理、病理中的地位,临证详辨肾中阴阳的各种变化,并以此为基础辨治妇科肾虚类疾病,创立养精育胞论。认为“三焦气化失司、心肾水火不济”是此类疾病的基本病机。首次提出以“温枢三焦,宁心坚肾”为治法,实现肾的阴阳平衡。基于养精育胞论的理论内涵,创制妇科调经1、2、3、4号方,构建中医妇科肾虚类疾病治疗新体系。并附验案加以阐明。

国医大师何成瑶养精育胞论治子宫腺肌病痛经经验

子宫腺肌病是一种良性、难治的妇科常见疾病,主要表现为月经异常、痛经、不孕等,而临床患者就诊多以痛经为首诊,检查发现患有子宫腺肌病。何成瑶教授从医60多年,对治疗子宫腺肌病有着丰富的经验及独到的见解,认为子宫腺肌病的病机为肾虚血瘀、水火不济、三焦不畅,根据其特殊的发病机理,首创了养精育胞的理论来治疗子宫腺肌病,在此理论指导下建立了温枢三焦、宁心坚肾的治法取得了很好的疗效。

哺乳动物精浆胞外囊泡及各组分的功能

精浆胞外囊泡是一种存在于精浆的膜性囊泡,按分泌器官分为附睾小体和前列腺小体。囊泡可与精子细胞膜发生融合,通过传递内容物或介导信号通路进而调节精子功能。它含有多种活性物质,其中蛋白质组分可影响精子活力以及顶体反应,并有清除损伤精子和促进细胞粘附的作用;脂质组分具有调节靶细胞质膜稳定性的作用;核酸组分主要参与免疫反应、跨代遗传及男性不育;离子则是多种酶的辅助因子,在调节酶活性和精浆微环境中发挥重要作用。不同组分对精子功能的影响不尽相同,本文将对此方面的研究进展进行详尽的综述,以期为该领域相关研究人员提供一定的参考。

Representative cDNA Library from Isolated Rice Sperm Cells

利用Percoll密度梯度离心和多次滤膜过滤分离到适于分子生物学研究的水稻 (OryzasativaL .cv .Guichao2 )生活精细胞。借助RT_PCR和SMART技术构建了水稻精细胞的cDNA文库。初级文库的大小为 1.5 8× 10 6 pfu ,插入片段平均大小为 980bp。 10 3个随机挑选的克隆与DIG标记的水稻幼苗根、叶及二细胞时期花粉、成熟花粉、精细胞和授粉 5～ 7d的子房cDNA探针杂交表明 ,除少数克隆外 ,它们在上述器官中的表达情况很相似。 10个随机挑选的克隆用来测序及分析 ,有 4个克隆含有完整的开放阅读框。 1个克隆与水稻Polyubiquitin (Rubq1)mRNA高度同源。Northern杂交表明它在二细胞时期花粉、成熟花粉中的表达显著少于在根、叶和授粉子房中的表达。另一个克隆的开放阅读框编码与拟南芥 (Arabidopsisthaliana (L .)Heynh .)AtRAD17同源的蛋白。这是第一次正式报道高等植物精细胞cDNA文库的构建 ,也是第一次从高等植物精细胞中分离到其表达的基因。

Molecular Cloning and Expression of RSSG58 Gene in Rice Sperm Cells

Myosins, a large family of structurally diverse mechanoenzymes, which, upon interaction with actin filaments, convert energy from ATP hydrolysis into mechanical force, play an important role in male reproductive processes. In this study we report the rice ( Oryza sativa L.) RSSG58 gene, which was cloned from the cDNA library of rice sperm cells by using sperm cell mainly expression subtractive clone as probe. This gene encodes a putative 66.7 W polypeptide, which shows similarity to the myosin heavy chain of Arabidopsis thaliana, and consists of 579 amino acids with an isoelectric point (pI) of 4.885. RSSG58, which is a member of a divergent gene family, generates transcripts of 2 278 bp and 2 437 bp that differ only in their polyadenylation sites. Southern hybridization showed that RSSG58 has only one copy in rice genome and RSSG58 transcripts are most abundant in sperm cells, with two distinct signals. The RT-PCR analysis indicated that the transcriptions of the RSSG58 gene were various in the different development stages and tissues. The greatest accumulation of RSSG58 mRNA was detected in sperm cells, while weaker expression was detected in leaves, microspore mother cells, unicellular microspore pollen stage, two-cell stage pollens, mature pollens and pollinated ovaries. These results suggest that RSSG58 is especially abundantly expressed in rice sperm cells.

Study on Key Technique of Artificial Insemination for Silkworm,Bombyx mori

[Objective] The aim of this study was to investigate the efficient technique of artificial insemination for silkworm. [Method] Sperms were extracted from bursa copulatrix of female moths mated for 30 min through extruding and centrifugal method, and then the semen was injected into other virgin moths with trypsinase. [ Result] A high-effective collection technology of spermatids from silkworm was established successfully, 50 μl semen could be collected by only one person in each hour. The survival rate of spermatids was over 80% in vito after collected from bursa copulatrix, while the obtained semen was quite pure and the average fertilization rate of silkworm was 76,5%. [ Conclusion] The establishment of high-effective semen extraction technique of silkworm provides the technical basis for studies on other related techniques for silkworm sperm.

F-Actin Visualization in Generative and Sperm Cells of Living Pollen of Rice Using a GFP-Mouse Talin Fusion Protein

Green fluorescent protein (GFP) fused to the F-actin binding domain of mouse talin labels the actin cytoskeleton in the living generative and sperm cells of a third generation transgenic rice (Oryza sativa L.) plant, A005-G-T-1-2. Observations were made on pollen at four major developmental stages, viz. I. uni-nucleate microspore stage; II. early bi-cellular pollen stage; III. late bi-cellular pollen stage; and IV. tri-cellular pollen stage. At each of these developmental stages vegetative nucleus, generative nucleus/ cell, and sperm cells were seen undergoing continuous and coordinated motion and migration. These movements seemed to be influenced by associated microfilament networks existing in the pollen. Based on these observations we propose that it is the interaction between the microfilament networks (usually one existing in the central cytoplasm and another in the cortex) that controls the dynamic movement of the vegetative nucleus, generative nucleus/cell and sperm cells. Furthermore, we have also observed that there is an array of microfilaments (oriented mostly parallel to the long axis of the cell) existing in the generative and sperm cells. As far as we are aware, this is the first report showing the existence of microfilaments in living generative and sperm cells of rice pollen. The implication and significance of the existence of microfilaments in generative and sperm cells in rendering self-propelled motion of these cells in relation to their passage and movement in the pollen tube and embryo sac for fertilization were discussed.

Preliminary Study on Transgenesis by Injecting Exogenous DNA into Zygote Cytoplasm of Buffalo

[Objective] This study aimed to investigate the feasibility of transgenesis by injecting exogenous DNA into zygote cytoplasm of Buffalo. [Method] Buffalo oocytes were randomly divided into two groups 20-22 h after in vitro maturation. One group of oocytes was introduced with about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection 7-10 h or 18-20 h after in vitro fertilization （IVF）; the other group of oocytes was introduced with mixture of a single buffalo sperm and about 7.5 pl of 50 μg/ml DNA solution containing linear EGFP fragment by cytoplasmic injection （generally called ICSI-Mediated Gene Transfer, ICSI-Tr）. Expression of exogenous DNA was observed and recorded during the process of embryonic development. [Result] Early embryonic gene expression efficiency and blastocyst gene expression efficiency in IVF injection group showed no significant difference compared with that in ICSI-Tr group （P0.05）. In addition, the cleavage rate and early embryonic gene expression efficiency in IVF injection group were significantly higher with injection at 7-10 h post IVF than that at 18-20 h post IVF （P0.05）. [Conclusion] These results indicate that transgenic buffalo embryos can be generated by injecting exogenous DNA into cytoplasm of IVF oocytes, and the optimal injection time is 7-10 h post IVF.

Synthesis and characterization of arginine-modified and europium-doped hydroxyapatite nanoparticle and its cell viability

The arginine-modified and europium-doped hydroxyapatite nanoparticles（HAP-Eu） were synthesized by hydrothermal synthesis.The prepared nanoparticles were characterized by transmission electron microscopy（TEM）,X-ray diffractometry（XRD）,Fourier transform infrared（FTIR） and zeta potential analyzer.The cell viability of HAP-Eu was tested by image flow cytometry.The results indicated that HAP-Eu is short column shapes and its size is approximately 100 nm,its zeta potential is about 30.10 mV at pH of 7.5,and shows no cytotoxicity in human epithelial cells and endothelial cells.

Effects of Cumulus Cells on in vitro Fertilization of Bovine

[Objective] The aim was to explore the effect of cumulus cells on the in vitro fertilization of in vitro matured bovine oocytes. [Method] The in vitro matured oocytes were divided into three groups of cumulus cells removal, partial removal and no removal. [Result] In the co-culture with cumulus cells, the oocytes of the removal group had higher cleavage rate and blastocyst rate （74.4%±4.1, 53.7%±5.1） than those of the no removal group （72.7%±5.1, 52.4%±3.5）, but the difference was not significant （P〉0.05）, while both groups had better performances than the re- moval group （39.6%±4.5, 18.8%±4.6） with the difference reaching the significant level （P〈0.05）. All the three groups showed significant difference with the control. The combination of cumulus cells and melatonin achieved the best effects as the cleavage rate and blastocyst rate of the partial removal group （79.8%±3.7, 56.5%±5.1） were better than those of the no removal group （78.2%±2.6, 55.8%±4.6）, and the difference was not significant, while both group had better performances than the removal group （48.3%±5.5, 22.7%±4.3） and the control group with the differences reaching the significant level （P〈0.05）. [Conclusion] The study provided technical support for the production of dairy cows and beef cattle.

Chronological and Morphological Progression of Nucleus during Mouse Oocyte Maturation and Fertilization in vitro

The chronological and morphological changes of the nucleus during mouse oocyte maturation and fertilization were systematically studied. Although most oocytes went through GVBD 2-4 hrs after culture, 13.6% remained at GV stage 8 hrs after culture.TEM observation revealed that nucleoli of oocytes which failed to go through GVBD were composed of fibrillar-granular component,small vacuoles and fibrillar centers or showed small vacuoles on nuclear surface. During GVBD, the nucleoli became smaller and smaller and finally disappeared with the nuclear-associated chromatin dislocated to the periphery. Nuclear membrane with attached chromatin became folded and electronic dense cores appeared in the center of chromatin clumps at the same time.The last event of GVBD was the disruption of nuclear membrane.At the end of the 5th hr after culture, meiosis progressed to prometaphase I.Chromosomes,distributed in the original GV area free of organelles,were surrounded by large quantity of mitochondria and small SER vesicles. At the end of the 12th hr after culture,48. 1% of the oocytes emitted PB1.Decondensing sperm head and early male pronuclcus(mPN)with condensed nucleoli were found 1-2 hrs after insemination.The formation and enlargement of female PN(fPN) occurred a little earlier than that of mPN. 33.3% finished syngamy at 8-9 hrs after insemination.The process of nucleolus formation was reverse to that in GVBD. The oolemma modification caused by cortical reaction could effectively inhibit polyspermy.in contrast,there were sperm binding to the oolemma where CGs failed to be released. In addition, PB2 was emitted 2-5 hrs after insemination. The difference between PB1 and PB2 as well as the abstriction of polar body were also discussed.

Morphological Characteristics of Smooth Muscle Cells Isolated from the Rat Ductus Deferens

The aim of this study was to establish a method of isolating and culturing smooth muscle cells from the ductus deferens of rats. Smooth muscle cells were prepared from ductus deferens by explanting technique after dissection of adventitia and intimae, and cultured in vitro. The identification of the smooth muscle cells were verified by using anti u-smooth muscle actin （a-SMA） immunohistochemistry studies. The result suggested that the cells are multi-morphous, showing long fusiform or star shapes. The apophysis of cells contacted and coalesced to each other, in some regions the cells overlapped in multilayer, while in the other regions they formed monolayer that fluctuated and showed a ＂peak-valley＂ shape. They presented a positive reaction through immunohistochemistry studies. The purity of the cells was more than 99% through this method. The culturing of smooth muscle cells by explanting technique is simple and stable.

Ultrastructure Features of Sperm Cell in Amaryllis vittata

Ultrastructure of sperm cells in pollen tube of Amaryllis vittata Ait. has been investigated in details by electron microscopy ,with particular emphasis on the organization and distribution of microtubules.The two newly formed sperm cells are arranged in tandem and sometimes in transverse at the right angle to the long axis of the pollen tube.Thevegetative nucleus is ahead and closely associated with the two sperm cells in all examined pollen tubes. The microtubules are distributed in the region between the common cell wall and the proximity of the sperm nucleus,they are singles and dispersed with mainly orientation of longitudinal and oblique,forming a loose bucket-like structure as a whole.In late stage of development,all of the microtubule array longitudinally and enclose the sperm nucleus.This configuration is very similar to the basket-like structure of microtubule in the generative cell. These results show that the organization and distribution of microtubules in the sperm cells are dynamic during cell development.

Cytological Mechanism of Cytoplasmic Inheritance in Pinus tabulaeformis: Ⅱ. Transmission of Male and Female Organelles During Fertilization and Proembryo Development

In an earlier report the ultrastructure and nucleoid organelles of male gamete in Pinus tabulaeformis Carr. have been described. Presently, the ultrastructure of the cytoplasm of the egg cell and pollen tube—immediately before fertilization and during cytoplasmic transmission of male gametophyte—has been described for the same species. The fate of parental plastids and mitochondria in the proembryo has also been followed. The mature egg cell contains a large amount of mitochondria, but seems to lack normal plastids. Most plastids have transformed into large inclusions. Apart from the large inclusions, there are abundant small inclusions and other organelles in the egg cell. During fertilization, pollen tube penetrates into the egg cell at the micropylar end and thereafter the contents are released. Plastid and mitochondrion of male origin are lacking near the fusing sperm_egg nuclei. The second sperm nucleus—not involved in karyogamy—remains at a site near the receptive vacuole. This nucleus is surrounded by large amount of male cytoplasm containing mixed organelles from the sperm cell, tube cell, and egg cell. At the free nuclear proembryo stage, organelles of male and female origin are visible in the perinucleus_cytoplasmic zone. Most of the mitochondria have the same morphological features as those in the egg cell. Some of the mitochondria appear to have originated from the sperm and tube cells. Plastids are most likely of male gametophyte origin because they have similar appearance as those of the sperm and tube cell. Large inclusions in the egg cell become vacuole_like. Paternal plastids have been incorporated into the neocytoplasm of the proembryo. In the cellular proembryo, maternal mitochondria are more abundant. Plastids resembling those of the sperm and tube cell are still present. These cytological results clearly show that in P. tabulaeformis, plastids are inherited paternally and mitochondria bipaternally. The cytological mechanism of plastid and mitochondrion inheritance in gymnosperm is discussed.