大鼠运动皮层投射神经元与Parvalbumin免疫阳性神经纤维终末的关系──免疫荧光、Confocal显微镜观察

Parvalburnin是细胞内一种钙结合蛋白。同时又可作为中枢神经系统内与GABA共存的神经元亚群的特异标记物，主要标记篮状及苔烛细胞。用PAP方谈染色可见大鼠Parvalbumin免疫阳性神经终末在运动皮层锥体神经元胞体周围形成包篮现象，但因该方法的局限性．较难明确二者的关系。为进一步了解Parvalbumin阳性终未在锥体神经元脑体、树突与轴突整体上的分布状况以及运幼皮层内不同传出神经元是否均接受同样的支配，本实验利用FastBlue送行标记、固定脑片细胞内注入LueiferYellow结合免疫荧光、Confocal显微镜观察，研究运动皮层内皮质丘脑（束旁核）、皮质效状体及皮质脊髓三种投射神经元与Parvalbumin阳性终末的关系。通过1．μ连续扫描图像的分析及立体对观察，Parvalbumin阳性终末清晰可见，与LuciferYellow标记的锥体细胞的关系也容易辨别．在三种投射神经元胞体上均可见Parvalbumin阳性终末包绕，形成明显包篮现象，但三种神经元上的终末数未见明显区别·阳性终未还分布于近端树突上，距胞体越远越稀疏：但在距脑体50μm以上的顶树突、30μm以上的基树突及其二、三级分枝的远端树突上仍偶有终末分布．此外，三种神经元轴突起始段上也有少量终末接触，但未形成明显的cartridge现象．这一结果揭示，Parvalb     

Oxygen Tension Imaging in the Mouse Retina

A newly developed microscope-based imaging system was used to measure the oxygen tension (PO₂) inside the retinal and choroidal vessels of mice and to generate in vivo maps of retinal PO₂. These maps were generated from the phosphorescence lifetimes of an injected palladium–porphyrin compound using a frequency-domain measurement. The system was fully calibrated and used to produce retinal PO₂ maps at different inspiratory oxygen fractions. PO₂ rose accordingly and predictably as inspiratory O₂ was stepped from hypoxic to hyperoxic conditions. Important experimental and acquisition parameters necessary for applying phosphorescence lifetime imaging to the mouse eye were investigated, including camera exposure and intensifier gain settings. Because of a need to limit light exposure to the retina, PO₂ map quality as measured by the coefficient of determination was investigated as a function of signal-to-noise and accumulated excitation energy deposition. With the development of this technology for use in mice, the potential for investigating the oxygen dynamics in genetically engineered mouse models of retinal disease, including diabetic retinopathy, glaucoma, and age-related macular degeneration, is advanced. © 2003 Biomedical Engineering Society. PAC2003: 4266Ew, 8763Lk, 8719Dd     

A 47,XXY fetus conceived after ICSI of spermatozoa from a patient with non-mosaic Klinefelter's syndrome: case report

The birth of 12 healthy infants to fathers with non-mosaic Klinefelter's syndrome has been reported so far. The spermatozoa for these pregnancies was obtained from frozen-thawed ejaculate in one pregnancy (twins) and from the testis in the remaining 10 infants. All of them had a normal karyotype. We describe a patient with non-mosaic Klinefelter's syndrome from whom a testicular biopsy was obtained and motile spermatozoa were collected. Of 16 oocytes that were injected, 14 fertilized and cleaved. Three embryos were transferred, resulting in a triplet pregnancy. Karyotype analysis from chorionic villous sampling revealed 46,XX, 46,XY and 46,XXY from the three fetuses. The affected 46,XXY fetus was reduced on the 14th gestational week. The pregnancy culminated with the birth of a healthy male and female, on the 36th gestational week, weighing 3600 and 2660 g respectively. This case report proves the presence of hyperploid spermatozoa in the seminiferous lumen, and strengthens the necessity of genetic diagnosis of the embryos or fetuses in such pregnancies to fathers with non-mosaic Klinefelter's syndrome.     

Fertility of ejaculated and testicular megalohead spermatozoa with intracytoplasmic sperm injection

In this study the fertility and outcome of intracytoplasmic sperm injection (ICSI) using megalohead spermatozoa from the ejaculates and testicles was evaluated. Seventeen males with megalohead and pinhead sperm forms in their ejaculate were studied in 22 cycles. A high number of sperm heads without tails and abundant round spermatid forms were commonly observed. Round-headed spermatozoa were seldom accompanied by these severely abnormal spermatozoa. The majority of megalohead spermatozoa were observed to have multiple tails, were predominant in the sample, and were used for ICSI. Ejaculated megalohead spermatozoa were used for ICSI in 15 cycles, while testicular spermatozoa were used in seven cycles where there were no vital spermatozoa or spermatozoa of low vitality in the ejaculate. The same abnormal morphology was observed in the testicles as in the ejaculated spermatozoa in the same males. Mean (+/- SD) low motility 4.7 +/- 5.6% and sperm count (3.8 +/- 4.19 x 10(6)) were common findings in these severely teratozoospermic patients. A low fertilization rate (43.2%) was achieved by using megalohead sperm forms (group I, n = 17) in comparison with the control group (60.2%) which had zero normal sperm morphology according to strict criteria (group II, n = 30) (P <0.01). Furthermore, a low pregnancy rate (9.1%) was obtained in the megalohead sperm group in comparison with the control group (40%) (P <0.05). Low fertilization and pregnancy rates may be due to a high incidence of chromosomal abnormalities from severely defective spermatozoa in the ejaculate. Couples should be counselled and warned about possible low fertilization and pregnancy rates with ICSI when only pinhead and megalohead forms with a high number of sperm heads without tails are present in the ejaculate.     

Effects of local nerve cooling on conduction in vagal fibres shed light upon respiratory reflexes in the rabbit

In ten vagus nerves the effect of local cooling on the compound action potential was studied in the temperature range of 34 to 0 °C in spontaneously breathing, anaesthetized rabbits. The mean temperature at which the myelinated (A) fibres were completely blocked, was 10.2±2.4 °C (mean ± S.D.). In nine nerves, local vagus cooling to 0 °C failed to block all non-myelinated (C) fibres. In one nerve, total blocking occurred at 2.0 °C. We conclude that in the rabbit, the earlier found increase in tonic activity of the diaphragm following lung inflation or deflation during bilateral local vagus cooling to a temperature between 8 and 0 °C is due to afferent impulses in vagal C fibres.     

腓浅神经痛诊治的解剖学基础(系列介绍之十五)

目的 介绍腓浅神经痛的诊断与局部注射的治疗方法。方法 选择门诊确诊为腓浅神经痛的患者18例，采用强的松龙混悬液加麻药局部注射神经穿出小腿深筋膜的部分的治疗方法。结果 所有病例皆在注射后短时完全止痛，多数病例一周后痊愈，少数病例经1～3次注射后痊愈。结论 对诊断明确的病例，采取强的松龙局部注射法，可以获得满意的疗效。     

Round spermatids from infertile men exhibit decreased protamine-1 and -2 mRNA

During spermiogenesis, histone-to-protamine exchange causes chromatin condensation. Spermatozoa from infertile men are known to exhibit an increased protamine-1 (PRM1) to protamine-2 (PRM2) protein ratio. Since patients undergoing testicular sperm extraction (TESE) followed by intracytoplasmic sperm injection (ICSI) reveal low fertilization rates, whether the outcome of ICSI could be related to the percentage of round spermatids expressing PRM1-mRNA and PRM2-mRNA was investigated. Applying in-situ hybridization, 55 testicular biopsies from men undergoing TESE/ICSI were investigated. The percentage of PRM1-mRNA and PRM2-mRNA positive spermatids was significantly (P < 0.0001) decreased in men with at least qualitatively normal spermatogenesis (PRM1-mRNA: 58.4 +/- 13.8%; PRM2-mRNA: 56.4 +/- 11.3%) and impaired spermatogenesis (PRM1-mRNA: 32.6 +/- 10.8%; PRM2-mRNA: 31.7 +/- 11.1%) compared with men with obstructive azoospermia and quantitatively normal spermatogenesis (PRM1-mRNA: 79.9 +/- 4.6%; PRM2-mRNA: 78.1 +/- 5.7%). A positive correlation (r(PRM1) = 0.733; r(PRM2) = 0.784; P < 0.001) was demonstrated between the score and the percentage of PRM1-mRNA and PRM2-mRNA positive spermatids. While successful fertilization was neither related to the score, nor to the percentage of PRM1-mRNA and PRM2-mRNA positive spermatids, a significant (P < 0.05) relationship was demonstrated between successful fertilization and the PRM1-mRNA to PRM2-mRNA ratio. Therefore, the PRM1-mRNA to PRM2-mRNA ratio in round spermatids may serve as a possible predictive factor for the outcome of ICSI.     

干扰素诱导淋巴瘤细胞凋亡实验及临床研究

目的：分析梯度浓度的干扰素(IFN-α)对Burkitt淋巴瘤细胞株Daudi和T细胞淋巴瘤细胞株Jurkut及15例难治性淋巴瘤患者的直接作用。方法：以MTT法测定梯度浓度的IFN-α对两种淋巴瘤细胞株Daudi、Jurkat增殖作用的影响，以DNA末端标记法，流式细胞术，电镜观察测定IFN-α对淋巴瘤细胞的凋亡诱导作用，并采用瘤内注射IFN-α联合化疗治疗15例耐药的难治性淋巴瘤。结果：低浓度亚IFN-α对DaudiJurkat细胞增殖无明显抑制作用，高浓度IFN(10000U／ml)可显著抑制两种细胞增殖，且有时间相关性。高浓度的IFN-α可诱导淋巴瘤细胞凋亡。15例患者CR5例，PR7例，有效率80％，无明显毒副作用。结论：IFN-α可抑制淋巴瘤细胞增殖，诱导凋亡，有显著时间，剂量依赖性。局部应用IFN-α联合化疗是治疗难治性淋巴瘤的有效方法之一。     

前臂后皮神经营养血管远端蒂复合瓣的应用解剖

目的:为前臂后皮神经营养血管远端蒂复合瓣设计提供解剖学基础。方法:30侧动脉灌注红色乳胶成人上肢标本,解剖观测前臂后皮神经营养血管的来源、分支、吻合及其与尺、桡骨膜血管的关系。结果:前臂后皮神经营养血管来自:桡侧副动脉皮支2～6支,外径(0.6±0.3) mm;骨间后动脉皮支6～9支,外径(0.7±0.3) mm;骨间前动脉腕背支皮支3~5支,外径(0.8±0.2) mm;尺、桡动脉腕背支皮支2~6支,外径(0.6±0.1) mm。尺骨中上段骨膜血管来自骨间后动脉的肌骨膜支6～8支,外径0.3～1.0 mm;骨间后动脉桡侧骨皮支与桡骨中段裸区骨膜血管吻合。上述支发出皮支、筋膜支、骨膜支和神经营养血管,形成皮神经干血管链以及深、浅筋膜和骨膜血管网。结论:前臂后皮神经营养血管与肌、骨、皮营养血管同源,其远端蒂复合瓣,旋转轴点在腕关节平面,适宜手背远处的组织缺损修复。     

Injection of substance P (SP) N-terminal fragment SP(1-7) into the ventral tegmental area modulates the levels of nucleus accumbens dopamine and dihydroxyphenylacetic acid in male rats during morphine withdrawal

The biologically active substance P (SP) N-terminal metabolite SP_1–7 has been reported to modulate several neural processes such as learning, locomotor activity and reaction to opioid withdrawal. Although all these processes are believed to be associated with dopaminergic transmission no evidence of an interaction between SP_1–7 and dopamine in the case of morphine withdrawal has so far been reported. Therefore, in this work we applied in vivo microdialysis to investigate the effect of SP_1–7 injection into the ventral tegmental area on dopamine release in nucleus accumbens of male rats during naloxone precipitated morphine withdrawal. The result showed that the heptapeptide enhances dopamine release and also elevates the level of the dopamine metabolite dihydroxyphenylacetic acid in this brain area. It was suggested that the observed action of the SP fragment on the dopamine system represents the underlying mechanism for a previously observed ability of SP_1–7 to counteract the aversion response to morphine withdrawal.