染色的

Granulosus is further;2. In differentiation of closely relatedspecies, the changes in heterochromatin and pericentric inversions are key factors, this is wellnoted in the small chromosomes;3. A. mantzorum has well-differentiated XY type sex chromo-'.omes, the Y chromosome is mainly composed of euchromatin, but has strong C-band in themiddle of the long arm. It is suggested that the Y chromosome of A. mantzorum is in the-initial stage of sex chromosome differentiation.

结果表明：(1)四川湍蛙、理县湍蛙和棕点湍蛙之间的亲缘关系较近，而它们与棘皮湍蛙的亲缘关系较远；2）在近缘种的分化中，染色体结构异染色质的变化和臂间倒位是重要的因素之一，这在小型染色体上表现得尤为突出；四川湍蛙具有在形态上分化很明显的性染色体。C带分析表明，此性染色体主要由常染色质构成，但在其Y染色体的长臂上存在明显的中间C带，推测尚处于性染色体分化的初期阶段。

The study showed the recombinant wt/mCREG protein depressed the VSMC proliferation depending on dose and the optimal concentration was 400nM;2biologic function of CREG protein and the membrance receptor mechanism:①effect on VSMC migration: the wound healing experiment showed the OB2 cells migration was slower significantly after added wt/mCREG(400Nm) in supernatant. The HITASY cells migration were very slowly and no remarkable change. The gelatinase digestion and Western blot analysis showed the matrix metalloproteinase was decreased and TIMPs was increased;②effect on differentiation: after added wt/mCREG(400nM), the expression of myocardin, SMα-actin, MHC and caldesmin were increased and that of LM-1 and FN were decreased in OB2 cells. These effects were more significant when adding wtCREG.;③effect on VSMC proliferation: Cell cycle assay and BrDU stain showed: after added the wtCREG and mCREG protein, the ratio of cell in G0/G1 phase increased to 0.5773 and 0.5572 from 0.5308 respectively in OB2 group, which increased to 0.7369 and 0.7034 respectively from 0.6297 in HITASY group;3Role of M6P/IGF2R in CREG biologic function:①ELISA and co-immunoprecipitation showed the wt/mCREG binding to M6P/IGF2R directly.②antibody blocking test: when the anti-IGF2R was added to medium at the same time with wt/mCREG at different concentration(2μg/mL、4μg/mL、8μg/mL),the effects of CREG protein which depressing proliferation, migration, secretion and promoting differentiation were blocked, which had the positive correlation to the concentration of added anti body. The studies showed two combinant CREG promoted VSMC switch to differentiation phaenotype, at the same time, depress VSMC proliferation, migration and secreting extracellular matrix.

上述实验结果证实：两种重组CREG蛋白对VSMC增殖均有剂量依赖性的抑制作用，并且相同浓度的糖基化的CREG蛋白对细胞增殖的抑制效应更为显著，最佳效应浓度为400nM；2两种重组CREG蛋白添加后对HITASY和OB2细胞生物学行为的影响：①CREG蛋白对VSMC迁移的影响：刮伤实验发现，加入最佳效应浓度的wtCREG和mCREG蛋白24h后，OB2组迁移能力下降，HITASY组无明显变化；细胞外基质金属蛋白酶-2，9(Matrix metallo-proteinase 2，9，MMP2 ，9)明胶酶电泳检测和Western blot检测结果证实，两种CREG蛋白均可以使OB2细胞合成细胞外基质MMP2，9减少，而组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases，TIMPs)增加；②CREG蛋白对VSMC分化的影响：加入400nM的wtCREG和mCREG蛋白12h后，OB2细胞myocardin、SMα-actin、MHC、caldesmin表达增加，LM-1、FN表达减少；③流式细胞仪分析细胞周期和BrDU染色分析证实，加入400nM的wtCREG和mCREG蛋白后，OB2组G0/G1期细胞由0.5308分别增加至0.5773和0.5572，HITASY组G0/G1期细胞由0.6297分别增加至0.7369和0.7034；3M6P/IGF2R在重组CREG蛋白的生物学功能中的调控作用：①免疫共沉淀和免疫荧光双染色分析结果显示，CREG蛋白与M6P/IGF2R存在直接结合；②应用抗体阻断实验：将不同浓度的anti- M6P/IGF2R(2、4、8μg /mL)与两种CREG蛋白同时加入培养液中，CREG蛋白抑制VSMC增值、迁移和合成细胞外基质、促进分化的效应减弱，而且与加入anti- M6P/IGF2R浓度正相关。

The growth inhibiting rate of T24 cell lines were detected by MTT methods, apoptosis of cells were detected by flow cytometry, the mechanism of apoptosis was analyzed by detecting the protein expression of Bcl-2, Bax, Caspase-9, Caspase-3 and cytoplastic protein Cytochrome C. 4 We injected live T24 cells into the subcutaneous space of nude mice and successfully built up the animal model of bladder carcinoma. The effect of CS-PAA-EPI polymer magnetic microspheres targeting chemotherapy was investigated by HE staining, TUNEL ,tumor weight and volume inhibition rate. Results: 1 TEM revealed that the CS-PAA polymer magnetic microspheres were regular spherical shape,the average diameter was 80nm in dry condition. By controlling the pH value of the medium,polymers had positive or negative zeta potential. VSM showed the CS-PAA polymer magnetic microspheres had superparamagnetic. The diameter of CS-PAA-EPI polymer magnetic microspheres were 200nm in solution by DLS examining,the embedding ratio was 20%,the EPI loading rate was 15%, which was higer than reported in other articles. 2 Raw eye observation found that the rat"s bladder of treatment group was brown color,which meaned the aggregation of iron particles, compared with the control group, iron stain found iron particles were assembled in rat"s bladder of the treatment group, the amount of iron particles in liver and spleen were less obviously.

研究结果：1合成的CS-PAA磁性聚合物微球呈球形，大小均一，TEM测定其干态下粒径为80nm左右，磁化曲线证实具有超顺磁性，具有一定的PH敏感性，固载表柔比星后，水溶液性状稳定，无沉淀物，DLS测定直径约200nm左右，测定载药率为15％，较文献报道高，包封率为20％。2肉眼观察试验组大鼠膀胱表面呈褐色，可见大量的Fe粒子聚集，普鲁士兰染色法显示，试验组大鼠膀胱壁内有大量的Fe粒子，分布至膀胱壁全层，与对照组大鼠相比，试验组大鼠的肝、脾内的Fe粒子聚集量明显降低；HPLC测定结果与Fe染色相同；高剂量磁性CS-PAA-EPI生理盐水组及单纯EPI生理盐水组均在给药后14天出现血肌酐和尿素氮的升高，其他组大鼠血生化指标没有明显变化。3MTT法发现，高、中、低剂量磁性CS-PAA-EPI生理盐水组在外加磁场的协同作用下杀伤T24细胞效应明显高于单纯的EPI生理盐水组，FCM发现试验药物组可引起明显的肿瘤细胞凋亡，试验药物治疗组细胞胞浆内出现了由线粒体释放出的细胞色素C，试验组细胞Bcl-2蛋白减少，Bax蛋白变化不明显，Caspase-3、Caspase-9蛋白受到了激活活化。4高、中、低剂量磁性CS-PAA-EPI生理盐水组的瘤重抑制率和瘤体积抑制率均明显高于单纯的EPI生理盐水组(P＜0.01)，其中高剂量组的抑制率最高。

Methods: Immunohistochemistry and image analysis techniques are applied to examine the phosphorylated extracellular signal－regulated kinase in epidermis of non－expanded skin and expanded skin. Results:Phosphorylated ERK expressed in basal cell layer of epiderm including non－expanded skin and expanded skin are expressed. But in expanded skin they are expressed significantly and densely compared to non－expanded skin. There are significant difference in relative to location of expanded skin and regions of expanded skin.

结果： 1扩张皮肤和正常皮肤表皮基底层都有磷酸化ERK 的表达和分布，但扩张皮肤中的磷酸化ERK的分布和表达较明显，染色较深且密，部分阳性细胞呈多层排列，有散在的增殖团区；2扩张皮肤的顶部和侧部差别不十分明显，而侧部和顶部与基底部有一定的差别，基底部染色较深，多为细胞核着色；3 经图像分析不同部位的正常皮肤和扩张皮肤以及扩张皮肤的不同部分的相对灰度值和阳性细胞密度，进一步证实了上述观察结果。

The signals look like noncontinuous beads on the DNA fiber,which consist of multi-copy and arrange tandemly.Fiber-FISH results with 45S rDNA probe showed that an average signal length is about 3～11μm in many tandem copy sequence(measure number,n=8),so we estimated the size of each copy to be approximately 11～30kb in cotton genome.There are dual signals on each end of the mitosis metaphase chromosomes and pachytene chromosomes hybridized with telomere DNA probe.The signals also look like non-continuous beads on the DNA fiber,the length is about 1～9μm,so we estimated the size is about 4～27kb.The signals almost covered the whole mitosis metaphase chromosomes and pachytene chromosomes of G.arboretum Shixiya 1 hybridized with gDNA probe,the euchromatin zone and heterochromatin zone were identified clearly on pachytene chromosomes,and the signals also look like non-continuous beads.Two BAC clones 150-D-24 and 182-F-9 in DNA BAC library of Pima90 were selected as probe to hybridize with mitosis metaphase chromosomes,pachytene chromosomes and DNA fiber of G.raimondii.

棉花FISH技术系统的应用研究。45S rDNA在亚洲棉石系亚1号中期和粗线期染色体上有两对大的信号，在DNA纤维上信号为非连续的念珠状，多个拷贝串联排列，每个拷贝长度大约在3～11μm，推测每个拷贝的实际长度为11～30kb；端粒序列在亚洲棉石系亚1号中期和粗线期染色体端部都有双点信号，在DNA纤维上信号也为非连续的念珠状，不同染色体上信号长度大约在1～9μm不等，推测实际长度为4～27kb；gDNA信号几乎布满整个亚洲棉石系亚1号中期和粗线期染色体，在粗线期染色体上能明显区分出常染色质区和异染色质区，DNA纤维上的信号均为非连续的念珠状。

Light microscope and transmission electron microscopy showed that SMMC-7721 cells induced by SAHA had undergone the restorational alteration in morphology and ultrastructure, which were different from those of nontreated cells but were similar to those of normal cells, and the changes were as follows: the cells turned to be flat and spread; the nucleo-cytoplasmic ratio lessened and nuclear shape became rather regular; the number of nucleolus reduced and its volume lessened; euchromatin increased while heterochromatin decreased in nucleus; in the cytoplasm, mitochondria grew in number with relatively consistent structure and well-developed mitochondria cristae; Golgi complex turned to be well-developed and typical; rough endoplasmic reticulum increased. Immunocytochemistry assay showed that the expression of AFP and PCNA were declined significantly. FCM analysis showed SAHA could arrest SMMC-7721 cells in G0/G1 phase, with an accumulation of the cells in G0/G1 phase while a decrease of cells in S phase. Semi-quantitative RT-PCR detection revealed that the expression of p21WAFl mRNA was upregulated remarkably in the cells treated with SAHA.

结果：倒置显微镜和透射电镜观察显示，经SAHA处理的细胞增殖速度显著减慢，细胞体积增大，细胞核较小，形状较为规则，核仁数量减少、体积变小，核内常染色质增多而异染色质减少，核质比例减小，细胞质内线粒体数量增多、线粒体嵴发达，高尔基体较为典型，粗糙型内质网增多，呈现出与正常上皮细胞相似的形态变化；MTT比色法测定结果显示不同浓度(2.5、5.0、7.5、10.0uM)SAHA对SMMC-7721细胞的增殖均有抑制作用，并有明显的剂量依赖和时间依赖关系；免疫细胞化学检测显示SAHA能显著降低PCNA和AFP在SMMC-7721细胞中的表达；流式细胞仪检测结果显示，SMMC-7721细胞经SAHA处理后，G0/G1期细胞明显增加，S期细胞则明显减少，细胞被阻滞于G0/G1期；RT-PCR检测结果表明，SAHA作用12h后SMMC-7721细胞中p21WAF1 mRNA的表达即有增加，24h后更为明显。

Each male ratwas paired with 2 female rats in proestrus when administration terminated. Femalerats were examined the next morning for the presence of sperm in vaginal smears andunderwent a cesarean section on day 13 of gestation. Then the reproductive indiceswere calculated as follows: copulation index, pregnancy index, and fertility index,which can evaluate male fertility directly. The sperm motility and morphology wereevaluated by computer-assisted sperm analysis, moreover, the caudaepididymal sperm ultramicrostructure were analyzed by transmission electronmicroscope, which can evaluate the influence on sperm maturation bydutasteride ; sperm survival rate was assessed by SYBR-14 and propidium iodidefluorescent staining; serumal dihydrotestosterone and testosterone of ratswere detected by enzyme-labeled immunoassay, which can evaluate the effecton androgenic levels; the weights of testes and epididymides, were determined byprecise electronic balance; histological examination of above tissues were evaluatedby HE staining and TEM, which can evaluate treatment on histology of reproductiveorgans by dutasteride.

给药结束后雄鼠与处于动情前期的雌鼠按2：1合笼，计算雌鼠的交配指数、受孕指数和生育指数，直接评价雄鼠生育力；采用计算机辅助精子分析系统分析大鼠附睾精子活力和形态，在此基础上合并使用透射电镜技术对各组附睾尾部精子进行透射电镜分析，评价度他雄胺对大鼠精子成熟的影响；采用SYBR-14和propidium iodide双重荧光染色计算精子存活率；采用Elisa法测定大鼠睾酮(testosterone， T)和双氢睾酮(dihydrotestosterone， DHT)血清浓度，评价度他雄胺对雄鼠雄激素水平的影响；采用精密电子天平对各组睾丸、附睾、前列腺和精囊进行称重，评价度他雄胺对大鼠生殖器官重量的影响；采用HE染色法对各组睾丸和附睾进行组织学分析，同时采用透射电镜技术对附睾上皮细胞超微结构进行分析，评价度他雄胺对大鼠主要生殖器官组织学的影响。

To understand the listed contents:the studies about component of nuclear pore complex; the main functions of gp210;the different configuration models of sequence specific DNA-binding proteins; karyotype and chromosome bandings; configuration of chromatin and dene transcription; nuclear bodies is many subnuclear domains which have different conformation between chromatin.

了解核孔复合体成分的研究；gp210主要的功能；序列特异性DNA结合蛋白的不同结构模式；核型与染色体显带；染色质结构与基因转录；染色质之间的空间发现的许多形态上不同的亚核结构域——核体。

Detecting each marker and cmparison the location of sequential tissuesection could determine the area loceation relationship of the markers, but it is difficulte to determine their correlation intra-cells exactly. No report of detecting virus genome and antigenes on a one section with triple staning techniques exists in the lit erature. in this report. by combining immunohistochemistry and in situ hybridization (with dioxigenin labelled HBV DNA probe), a triple staining technigce was established. With this technic, on one formalin-fixed and paraffin embeded section, HBV DNA, HBsAg and HBcAg were determinated one after the other.

各个细胞内病毒的复制和表达水平以及与肝细胞病变的关系，是乙型肝炎发病机理研究的一个重要方面：目前常用的在系列切片上对各标志物分别检测，比较观察的方法，一般只能大致了解各待检物在某一区域的定位关系，很难准确地了解它们在同一细胞内的关系：国内外尚未见有在同一张切片上三重标记染色检测病毒核酸及其多种抗原的报道：本文结合免疫组织化学ABC法及地高辛配基标记HBV DNA探针原位杂交，建立三重标记染色技术：在一张福尔马林固定石蜡包埋肝组织切片上先后检测，同时显示HBV DNA、HBcAg和HBsAg。

Histologically, the seminiferous tubules contained numerous Sertoli cells and more Sertoli-spermatozoa complexes, accompanied by the depletion of Leydig cells with deeply stained nuclei. Mature spermatozoa were stored up in the epididymis, but only a few in the efferent ducts. In the second place was testicular atrophy（32/120; 26.7%）. The seminiferous tubules showed moderate to severe inactivity of spermiogenesis with evidence of only spermatogonia, spermatocytes and Sertoli cells. The Leydig cells were obviously decreased in numbers associated with decrease of lipid droplets in their cytoplasms. Testicular hypoplasia was the third disorders（22/120; 18.3%）. Only a few spermatogonia and Sertoli cells appeared without any spermiogenesis. The associated changes was decreased in Leydig cells and fibrous hyperplasia in the interstitium.Epididymal stones were sometimes found（12/120; 10%）. Grossly, yellowish-white nodules with various sizes and firm in consistency were observed in the epididymis and the front efferent ducts. Microscopically, the epididymal ducts were dilated with voluminous spermatozoa storage, even showed calcification in severe cases. The deposited calcium salts were stained positively by Von Kossa and Alizarin red methods.Amyloidosis was also detected in 10 roosters（8.3%）. Eosinophilic, homogeneous, amyloid-like substances were deposited mainly in the testicular interstitium and the periphery of blood vessels. These substances showed positive reaction by Congo red staining. Five roosters（4.2%）had Marek's lesions in the testis, epididymis and peripheral nerves with infiltration of pleomorphic lymphocytes. Only one case showed epithelial necrosis of seminiferous tubules accompanied by fibrous proliferation in the interstitium.

结果发现，在总共搜集的120个病例中，其中因年老所导致的产精力不佳为最多，占38例（31.7%），於镜下可见大量精虫黏附於Sertoli cell的表面，并可见Sertoli cell数量明显增多而Leydig cell明显减少，且其细胞核呈现浓染的现象，而在其副睪中仍可见到成熟精虫蓄留於管腔中，但在其输精管内却只有少量精虫存在；其次为睪丸萎缩，占32例（26.7%），镜下可见中度至重度无造精作用，其生精小管中只见到精母细胞、精原细胞及Sertoli cell存在，但Leydig cell数量明显减少且其细胞质内的脂质也明显减少；睪丸发育不全，占22例（18.3%），於生精小管内只见到精母细胞及少量Sertoli cell存在，不见造精细胞分化，於生精小管间质可见Leydig cell减少并伴随结缔组织增生；副睪结石，占12例（10%），肉眼下可在副睪及输精管前段见到黄白色大小不一的结节，触感坚硬，於镜下可见副睪管扩张并有大量成熟精虫蓄积，严重时可见钙化现象，以Von Kossa及茜素红染色均呈阳性反应；类淀粉沉著症，占10例（8.3%），镜下在睪丸间质及血管周围可见粉红均质样的物质沉积，以刚果红染色成阳性反应；马立克病，占5例（4.2%），镜下可在睪丸、副睪实质及周边神经内均可见到嗜碱性大小不一的淋巴样细胞浸润；睪丸坏死，占1例（0.8%），镜下可见生精小管上皮细胞坏死脱落及间质结缔组织增生。

The labia have now been sutured together almost completely.The drains and the Foley catheter come out at the top.

此刻阴唇已经几乎完全的缝在一起了，排除多余淤血体液的管子和Foley导管从顶端冒出来。

To get the business done, I suggest we split the difference in price.

为了做成这笔生意，我建议我们在价格上大家各让一半。