灯盏细辛黄酮类成分对压力诱导的视网膜神经节细胞凋亡的影响

目的考察灯盏细辛中黄酮类成分(总黄酮、灯盏花素)对体外高压诱导的视网膜神经节细胞(retinal gan-glion cells,RGCs)凋亡的影响,探讨其视神经保护作用的物质基础。方法采用胰酶消化法将24只出生2~3d的SD(Sprague-Dawley)乳鼠视网膜制成细胞悬液,接种于经多聚鸟氨酸(HA)和层粘连蛋白(LN)包被的盖玻片中。培养72h后,将覆有细胞的血盖片转入加压装置中,加入黄酮类成分,继续培养48h,采用Fas蛋白免疫组化染色法及原位末端脱氧核苷酸转移酶标记(Tunel-POD)法进行检测,每天观察细胞形态,同时对部分细胞行NSE染色检查。结果细胞生长良好,NSE染色表明,85%以上的细胞为RGCs。给药组的Fas蛋白阳性表达指数和凋亡指数均明显低于模型组(P<0.05~0.01)。结论黄酮类成分均能对抗压力诱导的RGCs凋亡,为灯盏细辛视神经保护的有效组分。     

体外细胞培养检验“方证对应”关系的方法学研究

目的通过在体外细胞培养中加入中药复方粗提制剂对具有典型的中医证型的肺癌患者的外周血淋巴细胞NK活性的影响，探索能否在体外实验中反映“方证对应”关系。方法以水煎醇沉法制备两种中药复方（益气养阴和健脾化痰）的粗提制剂，取具有典型的中医证型（气阴两虚或脾虚痰湿证型）的肺癌患者的外周血淋巴细胞，将同一个患者的标本分为四个体外实验组，即空白对照组，中药5mg／ml组及10mg／ml组，自细胞介素2（250^u/ml，阳性对照）组。按“辨证论治”原则加入相应的中药复方制剂。体外培养22h或94h，以MTT法检测各组的NK活性。结果无论培养22h或94h，中药组的NK活性与对照组比较皆无显著性差异（P〉0．05），而白细胞介素2（阳性对照）组的NK括性均明显升高（P〈0．05）。结论两种中药复方制剂都未能在体外反应出“方证对应”的效果，说明中药粗提制剂直接用于体外细胞培养，其结果的可靠性差。     

Detection frequencies and the colony-forming unit recovery of oral treponemes by different cultivation methods

Selected media were compared for primary isolation and detection of oral treponemes from clinical samples. Forty-eight subgingival plaque samples from 45 patients suffering from periodontitis were anaerobically cultivated for 2 weeks at 37°C. Of the 9 media studied, Medium 10 (M10), which was supplemented with 10% rabbit serum and incubated using the plate-in-bottle method, supported the highest colony-forming units of the anaerobes. The treponemal colonies were detected at least on one medium from 83% of the subgingival plaque samples. The new oral spirochete medium in an anaerobic chamber supported the highest detection frequency of the oral treponemes (64% of samples); however, M10 in the plate-in-bottle was found to produce the highest colony-forming unit recovery of the oral treponemes (median 3.6% of the total colony-forming units). This study suggests that M10 in the plate-in-bottle and new oral spirochete medium in the anaerobic chamber are essential in cultivating oral treponemes.     

High Cell-Density Culture System of Hepatocytes Entrapped in a Three-Dimensional Hollow Fiber Module with Collagen Gel

Abstract: A compact three-dimensional (3D) module is needed for hepatocyte culture in order to develop an effective hybrid artificial liver system that can retain hepa-tocellular structure and differentiated functions. We treated the 3D module with collagen gel to entrap rat hepatocytes. This method yielded a high hepatocellular density (2 times 10⁷ cells/ml) over a period of 14 days and maintained the secretion of albumin and ureogenesis at the same level as the control monolayer method. The ammonia removal remained at 43% of the Day 0 value over 8 days of perfusion. Our data show that this approach may be useful for liver support therapy in an ex-tracorporeal circuit.     

Involvement of programmed cell death in preimplantation embryo demise

Fragmentation is frequently observed in animal and human embryosobtained via in-vitro fertilization (IVF), and is known to beassociated with decreased pregnancy rates and poor survivalfollowing cryopreservation. We postulate that embryo fragmentationis a consequence of activated programmed cell death (PCD) andsubsequent apoptosis and discuss evidence of morphological,histological and biochemical features compatible with the occurrenceof PCD in preimplantation embryos. If PCD is an underlying causeof the high incidence of the fragmentation seen in human pre-embryos,it remains to be determined whether this is reflective of thenatural incidence of lethal chromo somes in the human populationor due to the IVF procedure and culture conditions.     

人晶体上皮细胞的体外培养

目的:提供一种简单可行,易于掌握的体外培养人晶体上皮细胞的方法。方法:用无齿显微镊子将晶体囊膜分离出来,剪碎后直接移至细胞培养瓶中培养,当细胞长出融合后,用胰蛋白酶消化传代。结果:接种培养4天后,可见晶体上皮细胞开始长出,并以贴壁方式生长。结论:用此方法体外培养人晶体上皮细胞,具有简单、快捷、成功率高的优点。眼科学报1997;13:170—172。     

Protective Effect of Tetrandrine against Excitatory Amino Acids induced Neuronal Injury in Cortical Culture

兴奋性氨基酸类神经毒剂与粉防己碱（ｔｅｔｒａｎｄｒｉｎｅ，Ｔｅｔ）共同作用于原代培养胎鼠大脑皮层神经元２４小时，发现１０７，１０６ｍｏｌ·Ｌ１Ｔｅｔ明显降低５０μｍｏｌ·Ｌ１谷氨酸（ｇｌｕｔａｍａｔｅ，Ｇｌｕ），３００μｍｏｌ·Ｌ１βＮｍｅｔｈｙｌａｍｉｎｏＬａｌａｎｉｎｅ（ＢＭＡＡ，ＮＭＤＡ受体激动剂）和２０μｍｏｌ·Ｌ１βＮｏｘａｌｙｌａｍｉｎｏＬａｌａｎｉｎｅ（ＢＯＡＡ，ｎｏｎＮＭＤＡ受体激动剂）导致的培养液乳酸脱氢酶（ｌａｃｔａｔｅｄｅｈｙｄｒｏｇｅｎａｓｅ，ＬＤＨ）活性的增高；细胞形态损害减轻，细胞数量增加。对２０μｍｏｌ·Ｌ１ＮＭＤＡ介导的神经元损伤改变无影响。提示Ｔｅｔ对某些Ｇｌｕ类神经毒剂引起的胎鼠大脑皮层神经元损伤有一定保护作用，其机制可能是抑制细胞膜上的Ｎａ＋通道开放，阻止膜去极化而影响电压依赖性Ｃａ２＋通道启动。对ＮＭＤＡ受体可能亦有一定作用。     

The assessment of in vitro modulation of milk fat globule membrane expression by human breast carcinomas

Although knowledge of functional differentiation and tumour-associated changes of breast carcinomas can be gained by the application of antibodies directed against the milk fat globule membrane, more significant information may be obtained by assessment of the potential of breast carcinomas to modulate their antigenic phenotype. In this study, the extend to which primary tumours can undergo modulation in vitro has been investigated, with consideration of the suitability of organ culture in combination with the immunohistochemical detection of two milk fat globule membrane epitopes, HMFG1 and HMFG2, as methods for detecting this. The preservation of three of the 30 carcinomas assessed, all poorly differentiated, was poor after 3 days of culture. The viability of the other 27 was variable, and was greater in the better differentiated tumours and with the addition of insulin. Expression of the milk fat globule membrane epitopes was generally well maintained. Six of the carcinomas showed a significant change in antigen expression, with this being more frequent in tissues incubated with insulin. Hence, a small group of carcinomas have been identified which appear to have a greater capacity to undergo functional differentiation. Organ culture is considered to be a suitable method for maintaining the tissues in vitro for such evaluation, but the problems encountered in quantifying the immunohistochemical staining, because of antigenic heterogeneity, were such that it is suggested that other approaches be employed.     

在无血清培养基中代谢副产物对杂交瘤细胞生长代谢的影响 I.乳酸的作用

研究了无血清批培养中乳酸对杂交瘤细胞生长代谢的影响，当乳酸浓度低于5．3mmol/L时，乳酸对细胞生长和代谢无明显影响，当乳酸浓度在10mmol/L-25mmol/L时，对杂交瘤细胞的生长代谢有弱抑制，当乳酸浓度高于34mmol/L时，对细胞生长代谢有较明显的抑制。     

In search of the central respiratory neurons: II. Electrophysiologic studies of medullary fetal cells inherently sensitive to CO2 and low pH.

Although extensively pursued, the central respiratory neurons have remained elusive. We departed from the more conventional physiologic and morphologic methods of system and tissue examination and cultured dissociated fetal rat cells (Fitzgerald et al., J Neurosci Res 33:579-589, 1992) from the area of the nucleus ambiguus and the nucleus tractus solitarius located within the 2 mm rostral to the obex. Pacemaker-like cells, with a regular single or bursting activity, studied at 3-5 weeks of age, responded to very small pulses of CO2 (50 ms) and low pH with an increase in spike frequency and a decrease in spike amplitude. Other irregularly beating or silent cells did not respond or else required very large pulses (> 200 ms) to do so. The pacemaker cells also responded to hypoxia induced by administration of sodium hydrosulfite with an increase in spike frequency and amplitude; high oxygen (> 600 torr) and adenosine produced a decrease in electrical activity. Most of these cells were multipolar after staining with antibodies to neuron-specific enolase (NSE) and Fragment C of tetanus toxin. They did not stain for choline acetyltransferase (ChAT). The results suggest that these cultured cells, expressing a phenotype inherently responsive to CO2 and low pH, have the characteristics of central respiratory chemoreceptors, and may be involved in the generation of the respiratory rhythm.