AY245441基因在小鼠腭突发育和腭裂形成中的动态表达

背景与目的： 观察AY245441基因在正常腭组织和全反式视黄酸诱导的小鼠腭裂形成过程中的表达变化,探讨AY245441基因的表达在腭突正常发育和腭裂发生中的作用。 材料与方法： 利用Real-time PCR和原位杂交方法动态检测正常腭组织和全反式视黄酸诱导的腭裂组织中AY245441基因的表达情况。 结果： Real-time PCR结果表明,AY245441基因在GD11～16正常腭突和腭裂组织中均有表达,在正常腭中GD11,GD13和GD14时AY245441表达丰度最高,与其它胚龄相比较差异具有统计学意义(P<0.05);腭裂组中AY245441的表达丰度明显低于正常腭 (P<0.05),而在腭裂组织中各胚龄AY245441的表达丰度差异无统计学意义(P>0.05)。原位杂交结果显示,AY245441基因在GD13正常腭板前部的上皮细胞,GD14腭板前部上皮细胞和间充质细胞中均有表达 ;在GD14正常腭板后部上皮细胞中有少许表达,但间充质细胞中无表达。在GD13腭裂前部上皮细胞中表达量明显下降,在GD14腭裂前部上皮细胞和间充质细胞中表达受到明显抑制,几乎未检出阳性信号;在GD14腭裂后部上皮细胞和间充质细胞中均无表达。 结论： AY245441基因在小鼠腭突组织的生长发育中行使重要功能,与腭裂的发生有密切的关系。     

HSP47在小鼠腭裂和短肢发生过程中的表达

背景与目的:研究热休克蛋白47基因(Heat shock protein 47,HSP47)在小鼠胚胎腭、前肢正常和异常的发育过程中的表达情况。材料与方法:在GD10经口一次分别给予各实验组孕鼠80mg/kg的全反式视黄酸,对照组孕鼠给予等体积的植物油,并分别于GD11~GD18取两组胎鼠的前肢,于GD15~GD17取两组胎鼠的腭,利用RT_PCR方法半定量检测HSP47的表达丰度。结果:HSP47在所有样品中均有表达,其在GD11~GD18正常、异常发育肢的表达均呈随胚龄增大而增加的趋势,对照肢和实验肢分别于GD16、GD17达到高峰,以后基本恒定;其在GD15~GD17正常腭中恒定表达、异常腭中以GD16表达最高;异常肢的表达丰度在GD11~GD18均高于正常肢,正常腭和异常腭的表达丰度在GD15无差异,但在GD16~GD17异常腭的表达丰度则高于正常腭。结论:全反式视黄酸所致的短肢和腭裂中,HSP47的表达呈应激性升高。     

Hsp60基因在小鼠腭和肢正常与异常发生过程中的表达

背景与目的:研究热休克蛋白60基因(heat shock protein 60 gene,Hsp60)在小鼠胚胎腭、前肢正常和异常发育过程中的表达情况.材料与方法:将受孕后ICR小鼠随机分为实验组和对照组2组,每组各64只,于孕10 d(gestational day 10,GD10),分别经口一次给予实验组孕鼠80 mg/kg的全反式视黄酸,对照组孕鼠给予等体积的大豆油,并分别于GD11～GD18取2组胎鼠的前肢,于GD15～GD17取2组胎鼠的腭,利用实时荧光定量PCR检测Hsp60的表达丰度.结果:Hsp60在正常、异常肢和腭中均有表达.对照肢的表达丰度在GD14和出生前呈高表达,而实验肢的表达在各胚龄无明显差异(P＞0.05);实验肢Hsp60在GD11～GD18的表达水平高于同一胚龄的对照肢(P＜0.05).在正常腭中,Hsp60恒定表达,在异常腭中,Hsp60的表达随胚龄增大而降低;在GD15～GD17的表达丰度为实验腭低于同胚龄的对照腭(P＜0.05).结论:全反式视黄酸所致的短肢中,脚60的表达呈应激性升高;而在异常腭中,Hsp60的表达受到抑制.     

未折叠蛋白反应关键基因在小鼠前肢正常发育和异常发生过程中的表达

背景与目的： 研究未折叠蛋白反应(unfolded protein response,UPR)关键基因在小鼠胚胎前肢正常发育和异常发生过程中的表达情况。 材料与方法： ICR小鼠受孕后,将其随机分为实验组和对照组,每组各64只。于孕第10 d(gestational day 10,GD10),经口灌胃1次给予实验组孕鼠80 mg/kg的全反式视黄酸、对照组孕鼠给予等体积的大豆油,并分别于GD11～GD18取两组胎鼠的前肢,利用实时荧光定量PCR检测UPR关键基因Atf6、Ire1、Perk、Grp78的表达丰度。 结果： 除Ire1在GD18未被检出外,UPR中的上述4个主要基因在GD11～GD18正常肢发育过程中均有表达,且在GD13、GD17时间点前后呈现两个表达峰;其中Grp78的表达丰度最高,Ire1的表达丰度最低。而在实验组异常前肢发育过程中GD15之前上述4个基因的表达丰度均高于对照组正常前肢发育过程中的表达丰度(P<0.05);而GD15后均低于对照组正常前肢的表达丰度(P<0.05),呈相对稳定的低水平状态;且在GD12～GD14上述4个基因有一非常明显的表达高峰,其表达时程比对照组正常肢长,且表达丰度远高于对照组正常肢(P<0.05)。 结论： 全反式视黄酸所致的短肢模型中, UPR关键基因的表达在GD11～GD14显著升高,而在GD16～GD18则受到抑制,推测UPR可能与致畸有关。     

萘丁美酮的生殖毒性实验研究

本文选用小鼠睾丸生殖细胞染色体畸变分析试验，和大国发育毒性试验，对萘丁美酮的生殖毒性进行了研究，小鼠睾丸生殖细胞染色体分析试验设三个剂量组：１６７９、８４８．５、４２１，２５１ｍｇ／ｋｇ；和溶剂对照组。连续灌胃染毒５天，第六天取样分析。精原细胞染色体畸变细胞率、间隙率，初级精母细胞第一次减数分裂中期相的畸变细胞率和染色体早＊分离军，以及多倍体细胞率，与熔剂对照组相比较均无显著性差异，ＳＤ大凤的发育毒性试验设三个剂量组：２５０、６２．５和１５．６２５ｍｇ／ｋｇ，和一个敌枯双（１．６ｍｇ／ｋｇ）阳性对照组。分别于妊娠第７—１３天每日空腹灌胃染毒一次，并于妊娠第３、６、９、１２、１５．１８、２０和２１天称体重调整灌胃量，第２１天剖杀孕鼠取胎，观察并记录：孕鼠增重，死胎数，活胎数，胎重，胎鼠外观、内脏及骨骼畸形，以及胎鼠骨骼发育。结果表明：萘丁美酮对孕鼠增重，胎仔发育，胚胎及胎仔存活，胎仔骨骼发育均无影响；也未见引起胎鼠外观、内脏和骨骼畸形。本研究表明，萘丁美酮对雌性小鼠生殖细胞无遗传毒性，对大鼠胚胎及胎仔无发育毒性。     

五味子水浸提液对SD大鼠胚胎和胎仔的发育毒性研究

目的：观察五味子水浸提液对SD大鼠胚胎和胎仔的发育毒性。方法：采用SD孕鼠40只,分为溶剂对照组、五味子组,每组各20只。五味子组大鼠于妊娠第6天至妊娠第17天,经口灌胃给予五味子水浸提液（按每千克大鼠体质量15g生药）,每天1次;溶剂对照组孕鼠则给予等体积饮用水灌胃。实验过程中观察动物的体征变化,于妊娠第20天处死孕鼠并摘取卵巢和子宫,观察黄体数、着床数、胚胎生存及死亡情况、生存胎仔的外观、性别、体重、骨骼及脏器的发育等指标。结果：孕鼠及胚胎、胎仔发育的上述各观察指标未见明显异常,与对照组比较差异无统计学意义。结论：在本实验条件下,五味子水浸提液未见明显的母体毒性与胚胎和胎儿发育毒性。     

全反式视黄酸致ICR小鼠胎鼠短肢模型的建立

背景与目的：探索建立发生率高、畸形类型明确且易于获得的化学物致小鼠胎鼠短肢畸形的模型的方法。材料与方法：ICR小鼠合笼后,查到阴栓当天为孕期第0天（GDO）,孕鼠共50只,随机分为GD9、GD10、GD11、GD12给全反式视黄酸（all-trans-retinoicacid,atRA）实验组和对照组,共5组,每组10只,实验组经口灌胃1次给予孕鼠80mg／kgatRA,对照组则给予等体积的大豆油,于GD18将孕鼠处死并取出胎鼠,观察不同时间给予孕鼠atRA的胎鼠的短肢畸形情况。结果：于GD10给予孕鼠atRA,可致胎鼠双前肢短小,肱骨、桡骨、尺骨均较正常组短（P均〈0．05）;于GD11给予孕鼠atRA,可致胎鼠双前肢和双后肢短小,肱骨、桡骨、尺骨、股骨、胫骨均较正常组短（P均〈0．05）,腓骨常缺失;于GD12给予孕鼠atRA,可致胎鼠尺骨较正常组短（P〈0．05）。结论：成功建立了全反式视黄酸致小鼠短肢畸形的模型,为进一步研究肢体畸形的分子机制奠定了基础。     

小球藻片对大鼠的致畸试验

目的：评价小球藻片是否对SD大鼠具有致畸毒性,对其安全性进行毒理学评价。方法：孕鼠按体质量随机分为小球藻片低、中、高剂量组（分别为1 875、3 750、7 500 mg/kg,相当于人服用量的12.5、25、50倍）,阳性对照组（300 mg/kg阿司匹林）和阴性对照组（等体积纯净水）。于孕期7~16 d灌胃给予受试物,试验期间称量孕鼠体质量,第20天处死。剖腹检查子宫连胚胎质量、着床数、活胎数、死胎数、吸收胎数、胎质量和体长、胎仔雌雄比例、胎仔外观、骨骼和内脏发育情况。结果：阳性对照组大鼠具有明显的母体毒性、胚胎毒性和致畸毒性。低、中、高剂量组小球藻片无母体毒性、胚胎毒性和致畸毒性。高剂量组出现少数胎仔外观畸形和骨骼畸形,畸形率与阴性对照组比较差异无统计学意义（P > 0.05）。结论：在本实验条件下,小球藻片对SD大鼠无母体毒性、胚胎毒性和致畸毒性。     

乳酸钠对大鼠的致畸毒性

目的：检测乳酸钠对大鼠是否存在胚胎毒性和致畸毒性。方法：采用sD孕鼠,每组孕鼠〉12只。试验分为乳酸钠3个剂量组（750、1500和3000mg／kg）,阳性对照组（10mg／kg环磷酰胺）和溶剂对照组（双蒸水）,在胚胎器官形成期连续经口灌胃给予乳酸钠10d,妊娠第20d处死动物。剖腹取胎,检测乳酸钠对大鼠的母体毒性和胚胎毒性,并观察胎仔骨骼发育和内脏器官发育情况。结果：与溶剂对照组比较,阳性对照组出现活胎数减少、死胎及吸收胎数增多,胎仔身长、尾长、体质量减少,骨骼和内脏器官发育不全和畸形率升高等异常（P〈0．05）。除乳酸钠1500和3000mg／kg组在妊娠后期孕鼠体质量增长减慢,出现母体毒性外,其他各项指标,即黄体、着床、活胎、死胎和吸收胎的计数,子宫及胎仔的质量,身长和尾长的测量,内脏器官发育不全和畸形率,骨骼的骨化不全和畸形率等与溶剂对照组基本一致,均未见明显异常（P〉0．05）。结论：乳酸钠在〉1500mg／kg剂量时,对大鼠具有母体毒性,未见胚胎毒性和胎仔骨骼、内脏器官的致畸毒性。     

小鼠后肾集合管水通道蛋白表达及其上皮细胞超微结构研究

目的观察小鼠后肾集合管水通道蛋白(AQP)-4的表达及其上皮细胞超微结构变化。方法应用透射电镜、免疫组织化学及图像分析技术观察并检测小鼠后肾不同发育阶段集合管的超微结构及AQP-4表达。结果小鼠胚龄18 d见发育早期集合管主细胞,生后7 d～21 d,其形态结构发育基本完善。A型闰细胞于胚胎18 d出现,B型闰细胞生后21天出现。AQP-4于胚龄14 d始见表达,分布于集合管管壁上皮细胞侧基底细胞膜,随胚龄增加表达逐渐增强,生后1 d达高峰。结论集合管管壁上皮细胞于胚胎时出现,但其形态结构在生后才逐渐完善。AQP-4对小鼠胚胎时期肾水平衡调节可能起重要的作用。     

Factors inducing a loss of net negative surface charge on spleen cells of mice grafted with a slow-growing tumour.

The mean electrophoretic mobility (EPM) of splenic cells was determined in 4 different host-tumour systems. In splenic cells harvested from mice bearing slow-growing tumours, a significant EPM decrease was observed (12%) and in increase in the proportion of cells with slow mobility. Moreover, after 1 h incubation in RPMI medium at 37 degrees C and 2 washings, spleen cells showed a marked increase in their EPM (average 30%). Finally, the supernatant from incubation medium after contact with normal spleen cells (1 h at 37 degrees C) produced a significant decrease in their EPM (approximately 12%). On the other hand, no significant EPM variations were found between control spleen cells and cells from fast-growing tumours, before or after 1h incubation. The existence of factors which induce a loss of negative surface charge on spleen cells of some tumour-grafted mice is discussed.     

Wildervanck syndrome

We report a case of the Wildervanck (cervico-oculo-acoustic) syndrome exhibiting Klippel–Feil anomaly, congenital sensorineural deafness and bilateral sixth nerve palsy. Associated anomalies included short stature, microcephaly, mental retardation, and cleft palate.     

Partial functional overlap of the three ras genes in mouse embryonic development

In mammals, three ras genes, H-ras, N-ras and K-ras, encode homologous but distinct 21-kDa Ras proteins. We examined the in vivo functional relationship of the three ras genes in mouse embryonic development by investigating the phenotypes of mice deficient in one or multiple ras genes. H-ras-/- mice and N-ras-/- mice as well as a substantial proportion of H-ras-/-/N-ras-/- mice expressing only the K-ras gene were viable, while K-ras-/- mice were embryonically lethal, as have been reported previously. N-ras-/-/K-ras+/- mice died neonatally, while H-ras-/-/K-ras-/- embryos died much earlier than K-ras homozygous mutant fetuses. To further investigate the functional relationship of the ras genes in embryonic development, we introduced a human H-ras transgene into single or multiple ras mutant mice and found that the transgene rescued mice, including triple ras mutants, from embryonic lethality in association with correction of thin ventricular walls of the heart in null K-ras mutant mice. In situ hybridization revealed that the expression of the H-ras transgene on embryonic day E13.5 and E15.5 was more intense in major organs, including the heart, than those of endogenous ras genes. We therefore conclude that the functions of the ras genes are partially overlapping in mouse embryonic development.     

An Otorhinological study of patients with cleft lip and palate

32 patients of cleft palate with or without cleft lip were subjected to otorhinological study from January 1998 to September 2000, with special attention to the rhinological anomalies, ear pathology, any deafness, discharge or any other findings relevant to the middle ear. Hearing loss and ear changes were not seen in any patient with cleft lip alone, hence these patients were not included in this study. These changes were confined to the patients with cleft palate only, with or without cleft lip. High incidence of otological anomalies (75%) and rhinological anomalies (deviated nasal septum in 40.6%) were seen in patients with cleft palate with or without cleft lip. Hearing loss has been seen (37.5%) in patients with cleft palate and was purely conductive in nature.     

A Digital Cephalometric Study on The Morphometric Evaluation of Soft Palate in Oral Submucous Fibrosis

Objective: Oral submucous fibrosis (OSMF) is a chronic precancerous condition affecting the oral cavity, which is progressive and characterised by burning sensation and fibrotic change leading to restriction of mouth opening. This study evaluated the morphology of soft palate in different stages of OSMF patients using digital lateral cephalogram and compare it with healthy individuals. Methods: The study included 60 subjects, who were grouped as 30 OSMF and 30 healthy subjects from the same geographic population. Digital lateral cephalograms were taken with Planmeca Proline XC (Oy, Helsinki, Finland). Soft palate morphology was evaluated using Lateral Cephalogram, and the results were analysed statistically. Results: Leaf-shaped (Type 1) soft palate was commonly seen in the control group and stage I and II OSMF. Stage III OSMF patients presented with a butt-shaped (Type 3) soft palate. As the disease progressed, there was a conversion of Type 1 variety of soft palate to Type 3 variety. There was a gradual reduction in the length of the soft palate in the anteroposterior direction in OSMF patients compared to the control group. Conclusion: Early cephalometric diagnosis of soft palate changes may play a pivotal role in the overall management of OSMF.     

Immunotherapy of murine sarcoma by adoptive transfer of resident peritoneal macrophages proliferating in culture

Normal resident peritoneal macrophages from BALB/c mice were continuously grown and expanded in vitro as non tumorigenic cells on a confluent layer of mesothelial cells. These peritoneal macrophages expanded in vitro (EPM) were very cytotoxic against EMT6 sarcoma, Abelson myeloma, EL4, and L929S cells in culture. This tumoricidal effect was fully expressed without further activation with bacterial lipopolysaccharides (LPS). In vivo, adoptive transfer of one million EPM to BALB/c mice bearing subcutaneous EMT6 sarcoma caused regression of the solid tumor. In contrast, macrophages produced by 10 days' culture of bone marrow stem cells, or freshly isolated from the peritoneal cavity of BALB/c mice, were not cytotoxic in vitro or in vivo. Local injection in the vicinity of the tumor as well as intravenous transplantation of EPM effectively inhibited tumor growth. This antitumoral effect was further enhanced by intraperitoneal injection of 2 micrograms LPS to the tumor bearing mice.     

Cytotoxicity of subpopulations of splenic cells from normal and fibrosarcoma - bearing mice towards syngeneic tumour cells

The nonadherent splenic cells from normal and tumour-bearing (mouse fibrosarcoma-MFS) Swiss mice were divided into 6 subpopulations on Percoll step density gradient and characterised. For the determination of their cytotoxicity towards syngeneic MFS cells and their electrophoretic mobility (EPM), the splenic cell populations were pooled to form 2 broad groups: a lower-density group (density of saline to just less than 1.069 g/ml) and a higher-density group (1.069 to just less than 1.087 gm/ml). In general, the splenic cells from mice bearing 10- to 11-day-old MFS tumours differed in certain characteristics from those of normal mice in that they showed an increase in the following: proliferation, heterogeneity, with appearance of large cells (greater than 70 mu2); cells with a lower density (less than 1.069 g/ml); cells with a lower (less than 0.85 micron/sec/Volt/cm) anodi cEPM. The cytotoxicity studies revealed that: a) the lower-density splenic cells of both normal and tumour-bearing mice were more cytotoxic than the higher-density splenic cells; b) the lower- and higher-density splenic cells of tumour-bearing mice were more cytotoxic than the corresponding cells of normal mice. These findings indicate that the splenic cells of mice with a lower EPM and a lower density are the main contributors of cell-mediated cytolysis of a subpopulation of MFS cells.