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Setting: University hospital research laboratories Subjects and Interventions: NB4 and A549 cells were separately cultured with ATRA and/or dexsamethasone for 1-3 days. NB4 or ATRA -NB4 cells were then placed in an upper insert and co-incubated with A549 cells or their conditioned medium located in a lower plate.

材料与方法:将NB4 细胞与 A549 细胞分别经有/无全反式维甲酸及 dexamethasone 处理 1-3 天,再将有/无全反式维甲酸处理过之 NB4 细胞置入嵌入式培养之上层舱,下层舱中则置放有A549 细胞或各种不同处理的条件培养液,再将上层舱嵌入下层舱中,於含 5% CO2 之培养箱中共同培养。

In this paper, we studied M. meretrix ferritin, cathepsin B and caspase genes, which are involved in clam larval shell formation, nutrition, metabolism and apoptosis, respectively. We have cloned the three genes, investigated the temporal and spatial expression profile both at gene and protein level in trochophore (L1), D-veliger (L2), pediveliger (L3) and postlarvae (L4). The potential roles of these proteins were analyzed with specific inhibitors during larval development. Firstly, embryos were found developed into trochophore-like larvae with no shell if cultured at gastrula stage in artificial seawater without iron. Shell-like structures were formed only in the presence of iron. The larvae which had been transferred at L1 stage into ASW developed normal shell. This indicated that iron and iron associated protein are important for larval shell formation. The EST sequence which is homologous with ferritin, which is a principal iron metabolic protein, was selected from the M. meretrix cDNA library. The full-length of ferritin subunit cDNA was cloned by RACE. The results of real-time PCR revealed that the MmeFer mRNA expression changed before and after the larval shell formation.

本论文以文蛤幼虫为研究对象,分别对文蛤幼虫发育过程中贝壳形成相关的铁蛋白、营养及变态相关的组织蛋白酶B及变态过程中细胞凋亡相关的caspase三个基因进行了克隆,分析了基因及编码蛋白在担轮幼虫期(L1)、D形幼虫期(L2)、壳顶幼虫期(L3)和稚贝期(L4)的时空表达特征,解析了其可能的功能,并研究了相应酶类的特异性抑制剂作用对幼虫发育过程的影响,进行了目标蛋白的功能验证,详述如下:研究结果显示,在文蛤胚胎发育到原肠胚时放入不含铁离子的人工海水中培养,发育成无壳的畸形,随着人工海水中铁离子添加浓度的升高,幼虫长出壳状组织接近正常状态;而发育到L1期幼虫放入不含铁离子的人工海水中培养却可以发育出正常的壳,推测铁和铁代谢相关蛋白在幼虫贝壳初始形成有重要的作用。

And there were no evidence changes in histopathologic analysis of brain tissues in other ETU-injected and in control rat embryos. 2. Results of In situ cell apoptosis detection In E20 cerebral cortex, more Tunel cells were found in encephalocoele group than control group,and the results were significant.

二、应用细胞凋亡原位检测法检测不同组E20胎鼠大脑皮质中细胞凋亡情况 E20胎鼠大脑皮质中Tunel阳性细胞数目在正常组、给药无畸形组及脊柱裂组细胞凋亡数无明显改变,而在脑膨出组胎鼠大脑皮质细胞凋亡数明显多于对照组,计算凋亡细胞百分率,然后用X~2检验进行分析,脑膨出组和对照组的细胞凋亡指数差异有统计学意义(P.05)。

The biomechanical tests showed that two kinds of artificial bones had not significant difference on compressive strength and Young\'s modulus(P>0.05),while the flexural strength of nano-nacre artificial bone was less than the control group(P<0.05).3.The results of CCK-8 showed that the difference were not significant in each group,the proliferation of osteoblast reached the peak at the 5th day;7 days after being co-cultured,the total protein content of study group was higher than control group and blank group(P<0.05),while the difference between control group and blank group was not significantP>0.05The difference of alkaline phosphatase activities among three groups was not significant(P>0.05The SEM view showed that osteoblast attached and grew well in two kinds of artificial bone.4.X-ray photography showed that two kinds of powder started to degrade in 2 weeks;this phenomenon became more appear in 4 weeks,nano-nacre powder degraded faster than micron-nacre powder,while the hole shadow was easy to be found;in 8 weeks,all the femoral holes recovered and returned to normal bone mineral density in all groups.Analysis of tetracycline fluorescent double marks in the hard tissue grinding slices indicated that new bone grew fastest around the bone defect area in study group,while most slowly in blank groupP<0.05 SEM(scanning electron microscope observation showed that nano-nacre powder degraded more quickly.The same result can be found through the demineralized sections morphometric analysis,and both of the composite artificial bones made from those two kinds of nacre powder had the good connection with the adjacent tissue in rats body without apparent inflammatory response.5.X-ray photography showed that rabbit\'s bone defects healed faster in study group since NNAB implanted than in control group since MNAB implanted.At 24 weeks after operation,bone density in radial defects had nearly accessed to the normal area,while lower in control group,and turned up nonunion in blank group;The checking of BMD showed that results in study group were higher than those in control group at 8,16 and 24 week(P<0.05), and the difference between the BMD values in study group at 24 week and those in blank group was not significant(P>0.05).The gross specimens showed satisfactory histocompatibility both in study group and in control group,with bone tissue growing from two sides into the center of implanted materials; Normal slices in HE stain and hard tissue grinding slices in Stevenel\'s blue/Van Geison\'s picro-fuchsin stain showed that the bone growth tendency was better in study group than that in control group,and the medullary cavity had been penetrated to the implanted materials in study group at 24 week;Analysis of tetracycline fluorescent double marks in the hard tissue grinding slices indicated that new bone in both groups grew fastest 8 weeks after surgery,while slow down at 16 week.

纳米珍珠层/消旋聚乳酸复合人工骨与微米珍珠层/消旋聚乳酸复合人工骨分别与成骨细胞共培养后,其各时间点CCK-8法检测值与空白对照无显著差异(P>0.05),成骨细胞均在第5天达到增殖高峰期;培养7天后,实验组细胞蛋白含量高于对照组及空白组(P<0.05),后两者之间则无显著差异P>0.05碱性磷酸酶活性在三组间均无显著差异(P>0.05电镜下可见成骨细胞在两种人工骨上都有良好生长贴附能力。4.X-ray显示两种粉体在大鼠股骨骨洞植入第2周时都开始出现了降解,第4周时更为明显,纳米珍珠层粉较之微米珍珠层粉降解更快,而空白对照组骨洞阴影仍可见,至8周时,则所有组骨洞均己闭合修复,X-ray下已不可见原钻孔痕迹,恢复正常骨质密度;硬组织磨片四环素荧光双标记结果显示纳米珍珠层粉植入组较其余两组在骨缺损区周围新骨生长速度更快,空白组速度最慢P<0.05电镜观察及常规脱钙切片亦可见到纳米粉体降解较快;由以上两种原材料制得的纳米珍珠层/消旋聚乳酸复合人工骨与微米珍珠层/消旋聚乳酸复合人工骨在大鼠体内均与周围组织结合良好,无明显炎症反应。5.X-ray显示纳米珍珠层/消旋聚乳酸复合人工骨植入兔桡骨缺损区后其骨愈合速度较对照组微米珍珠层/消旋聚乳酸复合人工骨植入的快,至植入术后24周,实验组骨缺损区接近正常骨密度,对照组骨缺损区密度较低,空白组则呈现骨不连状态;骨密度测量结果显示术后8周、16周、24周实验组的骨密度值高于对照组(P<0.05,24周实验组的骨密度值与术前所测得的正常值无显著性差异P>0.05动物取材大体所见均显示组织相容性良好,骨组织逐渐由植入材料两端向中央生长;常规切片HE染色及硬组织磨片Stevenel\'s blue/Van Geison\'s picro-fuchsin联合染色均可见实验组骨缺损区长势优于对照组,至术后24周,实验组骨髓腔与材料已呈相交通状;硬组织磨片荧光显微镜下观察,两组材料在术后8周处于骨生长最快速时期,16周时速度开始减慢,术后4、8、16周时实验组的新骨生长速度均较对照组的快

Methods Six weeks old male Fischer 344 rats (F344) were used as donors, and six weeks old male F344alb as recipients. All F344alb were divided into 4 groups: group Ⅰ(n=10) was untreated; group Ⅱ(n=10) was treated with CCl4 but no γ-irradiation and transplantation; group Ⅲ(n=10) was transplanted with 1×10^7 BMCs after γ-irradiation, but without CCl4 treatment; group Ⅳ(n=10) was transplanted with 1×10^7 BMCs after γ-irradiation, with CCl4 treatment. The BMCs prepared from the donors were infused via the penial veins of recipients immediately after whole body γ-irradiation (7.5Gy) and CCl4 treatment: 4 weeks after BMCs transplantation, CCl4 were injected into peritoneal cavity twice a week for 4 weeks.

雄性F344大鼠为供体,而雄性F344alb为受体,分为4组:组Ⅰ(n=10)为正常对照组,不接受肝损处理,无全身射线照射和阴茎静脉骨髓细胞移植;组Ⅱ(n=10)为肝损无移植组,只接受肝损处理,无全身射线照射和阴茎静脉骨髓细胞移植;组Ⅲ(n=10)为正常移植组,全身射线照射后阴茎静脉移植骨髓细胞1×10^7;组Ⅳ(n=10)为肝损移植组,接受CCl4注射肝损,全身射线照射后阴茎静脉骨髓细胞移植1×10^7BMCs,其中,γ-照射剂量为7.5Gy,F344大鼠的BMCs经受体的阴茎静脉注入。

There were no statistically differences between the 8th~ 14thday, the 15th - 21th day post-BC and the control group. During the rt~24tkday post-BC , pycnosis degeneration or necrosis neurons of the locus cerebral cortex, dorsal hippocampus, dentate fornix were significantly increased, then decreased gradually, but recovered to normal by the 24thday after BC, and especially in parietal cortex and piriform cortex the necrosis neurons were significantly increased than temporal cortex , and there were no statistically difference berween the left and the right side. Pycnosis degeneration or necrosis neurons in the brainstem reticular formation were markedly increased in the 4thday after BC, and there were no statistically difference among the other groups and the control group.

与对照组相比,BC后在大脑皮质、背侧海马和齿状回部位,固缩变性和不完全坏死细胞数先显著增加(P<0.05),然后逐渐减少,至24d基本恢复正常;顶叶、梨状皮质比颞叶皮质变性坏死细胞多,差异有高度显著性(P<0.01);大脑左侧比右侧变性坏死细胞稍少但差异无显著性;在BC后4d,脑桥核、斜方体核平面的脑干网状结构中固缩变性和不完全坏死细胞数明显增多(P<0.05),其它组间无显著性差异。

There were no significant differences for the firing rates in the site of contralateral TNC neurons among during pre-CSD,CSD,and post-CSD (P>0.05).For flunarizine group,the firing rates in the site of ipsilateral TNC neurons during pre-CSD were higher as compared with during CSD(P<0.05).2.1 There were statistical differences on palasma levels of CGRP and SP among the three groups(P<0.05).The levels of CGRP and SP in CSD group were higher than control group(P<0.05).No significant differences on the levels of CGRP and SP in ipsilateral trigeminal ganglia were found among the three groups(P>0.05).2 The number of neurons with positive CGRP and SP immunoreactivity was statistically different in right-sided trigeminal ganglia among the three groups (P<0.05).The number in fight-sided trigeminal ganglia in CSD group was higher as compared with control group(P<0.05).The number in right-sided trigeminal ganglia was statistically higher than that in left-sided trigeminal ganglion in CSD group(P<0.05).3.1 Altered ReHo in ipsilateral pons and other brain regions response to pain such as basal nuclei,thalamus,cingulated gyms and prefrontal cortex was detected during the acute spontaneous attack as compared with during headache remission(P<0.05,corrected by Monte Carlo simulation). 2 Positive functional connectivity was detected between ipsilateral pons and other brain regions related to pain within pain state and within non-pain state (P<0.05,corrected by false discovery rate,FDR).Increased functional correlation between ipsilateral pons and other pain-related brain regions such as ipsilateral prefrontal cortex and contralateral subcallosal gyrus was detected during the acute spontaneous attack as compared with during headache remission(P<0.05,corrected by Monte Carlo simulation).

结果1。对照组未发现CSD;同侧TNC放电频率,CSD中>CSD后>CSD前P<0.05对侧TNC放电频率,CSD前、中、后无统计学差异(P>0.05氟桂利嗪组同侧TNC放电频率,CSD前>CSD中(P<0.05),CSD前与CSD后及CSD中与CSD后之间无统计学差异(P>0.05)。2.1关于放免测定,各组血浆CGRP、SP水平有统计学差异(P<0.05),CSD组高于对照组(P<0.05),CSD组与氟桂利嗪组、对照组与氟桂利嗪组之间均无统计学差异P>0.05各组之间同侧三叉神经节中CGRP、SP水平未见变化(P>0.05.2关于免疫组化研究,右侧三叉神经节CGRP、SP免疫阳性细胞数三组之间有统计学差异(P<0.05),多重两两比较结果CSD组大于对照组(P<0.05),CSD组与氟桂利嗪组之间、对照组与氟桂利嗪组之间无统计学差异P>0.05左侧三叉神经节CGRP、SP免疫阳性细胞数三组之间无统计学差异(P>0.05CSD组中右侧三叉神经节CGRP、SP免疫反应阳性细胞数大于左侧(P<0.05)。3.1局部一致性分析发现两组患者头痛疼痛状态较非疼痛状态脑活动发生变化的脑区有同侧脑桥以及其他疼痛相关脑区如基底节区、丘脑、扣带回、前额叶皮层等(P<0.05,蒙特卡罗模拟校正)。2功能连接分析发现疼痛状态与非疼痛状态下主要疼痛相关脑区均与同侧脑桥有功能联系P<0.05,false discovery rate,FDR校正疼痛状态与非疼痛状态比较,同侧前额叶皮层、对侧胼胝下回等疼痛相关脑区与同侧脑桥之间功能联系增强(P<0.05,蒙特卡罗模拟校正。

ANP and CX43 began to express at 2nd week after induction and increased gradually,about 60% of the resulting myogenic cells were positive at 4th week after induction ,they were negative for uninduced cells.hMSCs'surface antigen profiles obtained by Flow Cytometry were negative for CD31\CD34\CD45 before and after induction,but CD90 expressed higher after induction while was weak positive before induction(P.05). Apotosis index was correlated with the cultural time after induction,The apoptosis rate of induced hMSCs was remarkably higher than control group(P.01),and the variation between groups was notable(P.05),the cell cycle analysis showed that the percentages of G_0/G_1phases were reduced significantly after induction. The expresstion levels ofβ-MHC and CTNT mRNA were undetectable before induction,began to increase at 1st、4th week after induction,reached the peak at 6th week and decreased after that,the expression of Bcl-2 and Bax mRNA varied regularly after treated with 5-azacytidine. hMSCs'resting membrane potential、range and rate of depolarization were heightened gradually after being induced.

结果:hMSCs诱导前为纺锤形,诱导后第2天部分细胞即开始发生形变,呈球形或短棒状,1周后胞浆中颗粒增多,约20~30%细胞边缘呈毛刷样变化;hMSCs表面抗原CD31、CD34、CD45在诱导前后差异无统计学意义,CD90未诱导时表达呈弱阳性,诱导后明显增高(P.05);ANP和CX43在诱导前无表达,诱导后第2周开始表达且表达随时间逐渐增强,但CX43在诱导后第5周表达量开始降低。hMSCs诱导后凋亡指数随诱导后培养时间增加,低浓度诱导组低于标准浓度诱导组,组间差异有统计学意义(P.05),G_0/G_1期细胞比例诱导后较对照组显著减少(P.05);β-MHC和CTNT基因分别在诱导后第1周和第4周时表达开始增强,在第6周时均达到高峰,第8周时表达开始衰减,Bcl-2、Bax基因表达呈时间依赖性变化,hMSCs经诱导后随心肌样细胞特征的表达膜静息电位、去极化幅值和去极化速率逐渐增高。

The illumination used, so called soft x rays in the wavelength range of twenty to forty angstroms (an angstrom is one ten billionth of a meter), is also sufficiently penetrating to image intact biological cells in ma ny cases.

这就意味着生物样品可以在与它们自然环境相近的条件下被观察研究。其使用的照明度,即所谓的软性 X 射线,其波长在20到40埃之间(1 米的 100亿分之一为 1 埃)。在许多情况下也能够穿透完整无缺的生物细胞并成像。

The makers say that population pharmacokinetic analysis of schizophrenic patients who were enrolled in clinical trials showed that no significant pharmacokinetic interactions occurred with benztropine, lorazepam or propranolol. 4 The makers also point out that since ziprasidone is not metabolised by the cytochrome P450 isoenzyme CYP1A2, smoking should not affect its pharmacokinetics.

制造者们指出,对加入临床试验的精神分裂症患者的总体药代动力学分析表明,无显著的药代动力学相互作用发生在苯扎托品,劳拉西泮或普萘洛尔之间。4制造者还指出,由于齐拉西酮不是由细胞色素P45O同工酶CYP1A2代谢的,所以吸烟应该影响它的药代动力学。

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