小鼠精子微注射受精卵的体外发育与移植的研究

用显微注射方法,将精子注入小鼠卵母细胞卵周隙中,使精卵体外受精获得受精卵。并对精子注射后卵母细胞的成活率、受精率和受精卵的发育率以及移植后受胎率等方面进行了系统的研究,结果表明:卵子存活率为75.36％(1309/1737),其中264枚卵裂,受精率为20·17％(264/1309),卵裂卵经体外培养后有192枚发育到桑椹胚及胚泡期,发育率为72.73％(192/264)。将其中73枚(桑椹胚10枚,肛泡63枚)胚胎移植到10只假孕受体子宫中,一只受体妊娠产仔9只,仔鼠发育正常。     

刺激家兔小脑顶核对动脉血压和呼吸的影响

实验用家兔３７只，戊巴比妥钠麻醉，自然呼吸，其中２７只采用电刺激小脑项核，１０只为顶核内微量注射神经元胞体兴奋剂（Ｌ－谷氨酸钠）．观察到电刺激小脑顶核能引起动脉血压显著升高，但心电图记录无明显变化；呼吸运动明显加深加快，肺通气量显著增加．而项核内微量注射Ｌ－谷氨酸钠．则出现动脉血压降低，心率无明显变化，呼吸也无显著性变化．提示顶核神经元兴奋产全降压作用，而电刺激所致的升压和呼吸反应可能与顶核周围的神经纤维兴奋有关．     

Role of dicumyl peroxide on the morphology and mechanical performance of polypropylene random copolymer in microinjection molding

The purpose of this work is to systematically investigate the effects of dicumyl peroxide (DCP) on the microstructural evolution and mechanical properties of polypropylene random copolymers (PPRs) during the microinjection process. Polarized light microscopy, differential scanning calorimetry, X‐ray diffraction, and scanning electronic microscopy measurements were employed to characterize the morphology evolution of the PPR microparts with DCP. A hierarchical structure was found in the PPR microparts with DCP. Specifically, with the individual addition of organic peroxide, the orientation parameter of the PPR microparts decreased pronouncedly and the formation of skin layer was suppressed, while the formation of core layer was promoted. This was ascribed to the distribution of shear rate in the microchannel, which was determined by the wall ship effect in the filling stage and the relaxation behavior in the cooling stage. A mechanism was proposed to explain the distinctive filling behavior and molding characteristics of PPR with DCP in microinjection molding.     

青鳉鱼早期胚胎中外源DNA的复制行为的研究

利用显微注射技术将外源ＤＮＡ与标有生物素的ｄＡＴＰ一起导入处于１细胞期的青鱼（Ｍｅｄａｋａ）受精卵细胞质中，鱼卵在２５℃孵化一天至胚孔封闭期，从胚胎细胞质中提取ＤＮＡ，经分子杂交发现外源ＤＮＡ在胚胎中可进行复制。ＤＮＡ样品再经核酸电镜分析，发现了几种类型的复制分子，并发现多复制叉及成熟前复制现象。由此认为外源ＤＮＡ可以多种方式在青鱼早期胚胎细胞质中进行复制。     

On the origin of the “core‐free” morphology in microinjection‐molded HDPE

This study investigates the morphology of a high‐density polyethylene processed with microinjection molding. Previous work pointed out that a “core‐free” morphology exists for a micropart (150‐μm thick), contrasting with the well‐known “skin‐core” morphology of a conventional part (1.5‐mm thick). Local analyses are now conducted in every structural layer of these samples. Transmission electron microscopy observations reveal highly oriented crystalline lamellae perpendicular to the flow direction in the micropart. Image analysis also shows that lamellae are thinner. Wide‐angle X‐ray diffraction measurements using a microfocused beam highlight that highly oriented shish–kebab morphologies are found through the micropart thickness, with corresponding orientation function close to 0.8. For the macropart, quiescent crystallized morphologies are found with few oriented structures. Finally, the morphology within the micropart is more homogeneous, but the crystalline structures created are disturbed due to the combined effects of flow‐induced crystallization and thermal crystallization during processing. © 2011 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 49: 1470–1478, 2011     

Induction of Liver Size Reduction in Zebrafish Larvae by the Emerging Synthetic Cannabinoid 4F-MDMB-BINACA and Its Impact on Drug Metabolism

Zebrafish (ZF; Danio rerio) larvae have become a popular in vivo model in drug metabolism studies. Here, we investigated the metabolism of methyl 2-[1-(4-fluorobutyl)-1H-indazole-3-carboxamido]-3,3-dimethylbutanoate (4F-MDMB-BINACA) in ZF larvae after direct administration of the cannabinoid via microinjection, and we visualized the spatial distributions of the parent compound and its metabolites by mass spectrometry imaging (MSI). Furthermore, using genetically modified ZF larvae, the role of cannabinoid receptor type 1 (CB1) and type 2 (CB2) on drug metabolism was studied. Receptor-deficient ZF mutant larvae were created using morpholino oligonucleotides (MOs), and CB2-deficiency had a critical impact on liver development of ZF larva, leading to a significant reduction of liver size. A similar phenotype was observed when treating wild-type ZF larvae with 4F-MDMB-BINACA. Thus, we reasoned that the cannabinoid-induced impaired liver development might also influence its metabolic function. Studying the metabolism of two synthetic cannabinoids, 4F-MDMB-BINACA and methyl 2-(1-(5-fluoropentyl)-1H-pyrrolo[2,3-b]pyridine-3-carboxamido)-3,3-dimethylbutanoate (7′N-5F-ADB), revealed important insights into the in vivo metabolism of these compounds and the role of cannabinoid receptor binding.     

Germ cell transplantation in infertility mouse

This work investigated the spermatogenesis in an infertility BALB/c-nu mouse model by reinfusing germline stem cells into seminiferous tubules. Donor germ cells were isolated from male FVB/NJ-GFP trensgenic mice. Seminiferous tubule microinjection was applied to achieve intratubular germ cell transfer. The germ cells were injected into exposed testes of the infertility mice. We used green fluorescence and DNA analysis of donor cells from GFP transgenic mice as genetic marker. The natural mating and Southern blot methods were applied to analyze the effect of sperm cell transplantation and the sperm function after seminiferous tubule microinjection. The spermatogenesis was morphologically observed from the seminiferous tubules in 41/60 （68.33%） of the injected recipient mice using allogeneic donor cells. In the colonized testes, matured spermatozoa were seen in the lumen of the seminiferous tubules. In this research, BALB/c-nu infertility mouse model, the recipient animal, was used to avoid immunological rejection of donor cells, and germ cell transplantation was applied to overcome infertility caused by busulfan treatment. These results demonstrate that this technique of germ cell transplantation is of great use. Germ cell transplantation could be potentially valuable to oncological patients.     

利用TALEN技术制备HE4（WFDC2）敲除小鼠模型

目的 利用显微注射技术和 TALEN技术构建HE4（WFDC2）敲除小鼠并对其进行表型分析。方法 设计Wfdc2敲除位点,构建TALEN载体,体外转录TALENs获得mRNA并通过显微注射技术注射到C57BL/6 J小鼠受精卵,对F0代小鼠进行DNA鉴定获得HE4（WFDC2）敲除小鼠,观察并统计 HE4（WFDC2）敲除小鼠出生及存活情况。结果 在Wfdc2第一个外显子上设计了TALEN识别剪切位点,向受精卵注射TALENs mRNA获得了24只F0代小鼠,其中1只发生移码突变,成功制备了HE4（WFDC2）敲除小鼠,并发现纯合 HE4敲除小鼠出生后在短时间内致死。结论 通过TALEN技术成功制备HE4（WFDC2）敲除小鼠,纯合HE4（WFDC2）敲除小鼠出生致死。