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MethodsNeural tubes isolated from the region between midotic placode and third somite of mouse embryo at ED 8.5d were cultured in DMEM/F12 serum-free medium containing basic fibroblast growth factor and epidermal growth factor. The cell cycle of CNCCs was analyzed with flow cytometry, the proliferation capability was determined by MTT method, AP-2α expression was detected by immunocytochemistry as a biological marker of cardiac neural crest cells. The features of migration, multi-directional differentiation were observed as well.

材料与方法成年2月龄C57/BL6小鼠交配,取ED8.5d小鼠胚胎枕中部至第3体节神经管,应用包含碱性纤维生长因子和表皮细胞生长因子的无血清培养基培养,利用MTT法检测细胞增殖能力,利用流式细胞仪检测细胞周期,采用转录激活因子2α(AP-2α,Activator protein-2α)作为其生物学标记物,观察其增殖状况、细胞周期、迁移、分化等生物学特性。

RESULT: 1.Ouabain act on lens sodium-pump,under the LM,lens anterior capsular membrane discontinuous,epithelium cells clustered,occluding zonule seperated,lens fiber layers fractured.Under the EM,cells totally hollowed,mitochondria swelling,myelin figure appeared.RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1、α2 and α3-isoform are all decreased.2.Digoxin act on lens sodium-pump,under the LM,lens cell oedema,linkage distructed,extensive exfoliation.Under the EM,plasma appeared little half-transparant hollow region,mitochondria swelling and ridge disappeared. RT-PCR examine,α1、α2 and α3-isoform are all decreased.3.Amphotericin B act on lens sodium-pump,under the LM,lens epithelium cells linked tightly,arranged in-line,lens fiber layers arranged tightly and regularily.Under the EM,abbundant cellular organes,exuberant cells function indicated. RT-PCR examine the expression condition of αsubunit of sodium pump on mRNA level,α1 and α3-isoform are increased significantly,demonstrated isoform-specific action.4D-thyroxine act on lens sodium-pump,under the LM,lens plasmalemma integrated,cells arranged tightly and regularily.Under the EM,nucleus fission appeared,desmosome half-desmosome and tensile microfilaments linked the cells. RT-PCR examine,α2 and α3-isoform are increased, also demonstrated isoform-specific action.5.Vitamin E act on lens sodium-pump,under the LM,lens anterior capsular membrane continuous and smooth,epithelium cells tightly linked,lens fiber layers appearede hollow region occasionally.Under the EM,lateral membrane high density belt appeared,abundant nucleolus. RT-PCR examine,onlyα1-isoform are increased, demonstrated significantly isoform-specific action.6.DMSO act on lens sodium-pump,under the LM,lens anterior capsular membrane slightly thicker,cells linkage partly distructed.Under the EM,plasmalemma denaturation,mitochondria swelling.RT-PCR examine,α1、α2 and α3-isoform are all altered slightly and haven't significant meanning.

结果:1、哇巴因作用于晶状体钠泵后,光镜下晶状体前囊膜断裂、上皮细胞聚积、闭合连接分离、纤维板层裂隙,电镜下全层细胞空泡化、线粒体肿胀出现髓样结构,RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达均减弱。2、地高辛作用于晶状体钠泵后,光镜下晶状体细胞水肿、细胞连接破坏、广泛剥离,电镜下胞质见少许半透明空化区、线粒体肿胀嵴消失,RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达均减弱。3、两性霉素B作用于晶状体钠泵后,光镜下晶状体上皮细胞紧密连接、线状排列、纤维板层紧密规整,电镜下细胞器丰富、细胞功能旺盛,RT-PCR法检测α1及α3表达显著增强、具有一定的重整异构作用特异性。4、D甲状腺素作用于晶状体钠泵后,光镜下晶状体质膜完整、细胞排列紧密规整,电镜下胞核见分裂像、细胞间有桥粒、半桥粒及张力微丝,RT-PCR法检测α2及α3表达均增强、亦有一定的重整异构作用特异性。5、维生素E作用于晶状体钠泵后,光镜下晶状体前囊膜连续光滑、上皮细胞紧密连接、纤维板层偶见空化,电镜下囊侧膜内有高电子密度带、细胞核仁丰富,RT-PCR法检测仅有α1的表达显著增强、具有极强的重整异构作用特异性。6、二甲基亚砜作用于晶状体钠泵后,光镜下晶状体前囊膜轻度增厚、细胞连接部分破坏,电镜下质膜变性、线粒体肿胀,RT-PCR法检测晶状体钠泵α亚单位α1、α2及α3三种重整异构体在mRNA水平的表达无显著改变。

N perifocal tissue intracerebral hemorrhage there were rarefaction neuron,cell spaces augmentation,cell diminution,distinct demarcation of cell membrane and surrounding,and we discovered a lot of degeneration and necrosis nerve cells,cell body collapsed,pycnosis anachromasis,nucleoli disappeared. In EPO group we discovered that center area of hemorrhage shinked,nerve cells of degeneration and necrosis decreased in perifocal tissue,majority cells morphous were normal and pathological changes were light.

CH对照组在术后皮层出血中心无神经元,仅见少量胶质细胞,细胞间质空泡样改变;出血边缘区神经元稀疏,细胞间隙增大,细胞缩小,胞膜与周围分界清楚,并可见大量变性及坏死的神经细胞,表现为胞体皱缩,核固缩深染,核仁消失。rhEPO治疗组出血中心区变小,边缘区神经细胞变性坏死减少,多数存活细胞形态相对正常,病变较轻。

Results: NKA-I cells was first found in the duodenum mucose on the 14th day of fetus, earlier than in the jejunum ( 16th day ) and ileum ( 17th day ), and its Positive reaction and the number of cells increased with further development, after birth the 30th day, it became quite identical to that of adult rat; NKA-I cells were seen in the colon mucosa on the 17th day of fetus, later on the number kept decreasing till grown up stage; No change was observed in the stomach mucosa except that few cells showed up on the 15th day of fetus; No positive cell was found on the oesphagus wall.

结果: NKA-I细胞首先见于胚胎14 d的十二指肠粘膜,早于空肠(胚胎16 d)和回肠(胚胎17 d),其阳性反应强度及细胞量随发育而增加,30 d时与成年鼠相似;结肠粘膜内于胚胎17 d见细胞,但此后数量较少直至成年;胃粘膜于胚胎15 d见个别细胞外,以后无明显变化;食道壁始终未见阳性细胞。

Results: NKA-I cells was first found in the duodenum mucose on the 14th day of fetus, earlier than in the jejunum ( 16th day ) and ileum ( 17th day ), and its Positive reaction and the number of cells increased with further development, after birth the 30th day, it became quite identical to that of adult rat; NKA-I cells were seen in the colon mucosa on the 17th day of fetus, later on the number kept decreasing till grown up stage; No change was observed in the stomach mucosa except that few cells showed up on the 15th day of fetus; No positive cell was found on the oesphagus wall.

结果:NKA-I细胞首先见于胚胎14 d的十二指肠粘膜,早于空肠(胚胎16 d)和回肠(胚胎17 d),其阳性反应强度及细胞数量随发育而增加,30 d时与成年鼠相似;结肠粘膜内于胚胎17 d见多个细胞,但此后数量较少直至成年;胃粘膜于胚胎15 d见个别细胞外,以后无明显变化;食道壁始终未见阳性细胞。

Results Myoid/myofibroblastic differentiation occurred most commonly in fibrosarcomatous DFSP. It was recognized histologically as peripherally distributed or randomly scattered small eosinophilic nodules or short bundles, which were composed of bland spindle cells, closely resembling smooth muscle cells or myofibroblasts.

结果 肌样/肌纤维母细胞性分化多出现在纤维肉瘤型DFSP中,表现为肿瘤周边部或肿瘤内散在性分布的深嗜伊红色小结节或短条束,由梭形细胞组成,细胞多无异型性,核分裂象也罕见,形态上似平滑肌细胞或肌纤维母细胞。

The expression of subunits α5,β1,β3 was detected in normal morulas and increased in blastocysts. The expression of subunit α1 was not detected in morulas or blastocysts, but detected only in outgrowing trophocytes. 3. Expression of integrin β3 in irradiated group embryos was weaker than that of normal group. There was no expression of integrin α1 both in morulas and blastocysts in irradiated group and no difference in the expression of subunits α5 and β1 between these two groups.

结果:1、超声波照射组胚胎贴附率、滋养细胞外延生长率均显著低于正常胚胎。2、正常胚胎桑椹胚期即有α5、β1、β3整合素表达,而无α1表达;囊胚期α5、β1、β3表达增强,仍无α1表达;α1只出现在外延生长的滋养细胞。3、超声波照射组胚胎β3整合素的表达明显弱于正常组,且未见α1表达;而α5、β1的表达两组间无差异。

The survival rates of CEM cells cultured with cis-diamminedichicloroplatinum added 24 h later were higher than that cultured with hTERT ASODN and DDP added 24 h later. The survival rates of CEM cells cultured with DDP were similar with that cultured with hTERT SOND and DDP. In morphological observation of apoptotic cells using Giemsa staining, cells treated with DDP or DDP combined with hTERT ASODN ro SODN at 48 h, displayed classic apoptotic changes. Apoptosis rates of CEM cells treated with DDP for 48 h after 24 h of exposure to ASODN significantly increased. There was significant difference in the percentage of apoptotic cells of CEM cells between hTERT ASODN plus DDP and SODN plus DDP or DDP alone, respectively.

结果: hTERT ASODN作用于CEM细胞24 h再加入柔红霉素、长春新碱、足叶乙甙,对细胞生长的抑制分别与单用柔红霉素、长春新碱、足叶乙甙及hTERT正义寡核苷酸联合柔红霉素、长春新碱、足叶乙甙组相比,统计学上无显著差异(P>0.05)。hTERT反义核酸作用于CEM细胞24 h加入顺铂,再共同作用48 h,CEM活细胞均数为2.318×108 cells/L,与单用顺铂组(3.250×108 cells/L)及hTERT正义核酸联用顺铂组(3.175×108 cells/L)相比,对细胞抑制明显增强(P<0.05)。hTERT ASODN作用于CEM细胞24 h再加入顺铂作用48 h,细胞出现典型的凋亡形态学改变。hTERT ASODN与2.5 μmol/L顺铂联合作用于CEM细胞48 h的凋亡细胞百分率(19.47%)分别同SODN与顺铂联合作用组(6.97%)、单用顺铂作用组(6.02%)进行比较有显著差异(P<0.01)。

Before the two-hour interaction of male-male individuals , we found there were obviously fewer and lighter stained ERp immunoreactive neurons (ER|3-IRs) in both main olfactory bulb system project regions and vomeronasal system project regions in the mandarin voles than in the reed voles, but no significant difference in numbers of AR immunoreactive neurons between the two species through the examining of immunoreactive neurons, moreover, there were significantly fewer AR-IRs than ERp-IRs in two species of vole before the two-hour interactions.

免疫组织化学检测,没有社会交往时棕色田鼠主嗅球系统投射区和犁鼻系统投射区ERβ免疫阳性细胞明显少于沼泽田鼠,且显色淡,AR免疫阳性细胞在两种鼠间差异不大,且都明显少于各自的ERβ-IRs.2h交往后,棕色田鼠主嗅球投射区和犁鼻系统投射区的ERβ-IRs细胞数明显少于交往前,AR-IRs细胞数明显多于交往前;沼泽田鼠交往前与交往后ERβ-IRs和AR-IRs细胞数均无显著差异,且显著多于交往后棕色田鼠ERβ-IRs细胞数,显著少于交往后棕色田鼠AR-IRs细胞数。

Results Positive staining for VEGF was distributed mostly in osteoblasts, osteoclasts and newly-born osteocytes. Vascular endothelial cells and bone marrow cells also showed VEGF positive reaction. Hypertrophic chondrocytes in calcifying region of chondroephysis were VEGF positive, but VEGF was absent from chondrocytes in other zones of chondroephysis.

结果 VEGF阳性反应主要见于成骨细胞、破骨细胞及新生骨细胞中,血管内皮细胞及骨髓细胞也呈阳性反应;骺软骨钙化区肥大的软骨细胞为VEGF阳性反应,但其它区域的软骨细胞中无VEGF表达。

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