Mechanisms of changes to the liver pigmentary component during the annual cycle (activity and hibernation) of Rana esculenta L.

The present study was performed to elucidate the mechanisms responsible for the changes of melanin content/distribution we had previously discovered in the liver parenchyma of Rana esculenta during natural hibernation. Melanomacrophagic component response was analysed using morphocytochemical methods. The results demonstrated that during the prehibernation period (October–November) the melanomacrophages reach the highest proliferative activity (BrdU, PCNA labelling) which is accompanied by an evident melanosynthesis (dopa‐oxidase activity). In contrast, after hibernation, the decrease of liver pigmentation was the consequence of a partial cell loss by apoptotic mechanisms (TUNEL labelling, pyknosis‐karyorhexis) accompanied by a decrease of melanosome content by autophagy and low melanosynthetic activity. On the basis of these findings, there is evidence that liver melanomacrophages represent a metabolically (melanin synthesis/degradation) and cytokinetically (proliferation/death) active cell population during the annual cycle of the frog. The results are also discussed in relation to the functional synergism between hepatocytes and pigment cells in the adaptation to environmental changes.     

Cell death and cell proliferation in cartilage layers in human anterior cruciate ligament tibial insertions after rupture

The purpose of this study is to investigate cellular responses and histological changes of cartilaginous layers in human anterior cruciate ligament (ACL) tibial insertions after rupture compared with those in normal insertions. Fully 16 tibial insertions of ruptured ACLs were obtained during primary ACL reconstructions. We also obtained 16 normal ACL tibial insertions from cadavers. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) to detect apoptosis, proliferating cell nuclear antigen (PCNA) staining, and histological examination were performed. The percentage of TUNEL-positive chondrocytes in ruptured ACL insertions (30.2 ± 15.6%) was higher than that in normal insertions (9.6 ± 5.8%). The percentage of PCNA-positive chondrocytes was significantly different between ruptured ACL insertions (19.9 ± 15.0%) and normal insertions (12.3 ± 7.3%). The average thickness of the cartilage layer, the glycosaminoglycan-stained area, and the number of chondrocytes per millimeter in ruptured ACL insertions was smaller than those in normal insertions. The decrease in the number of chondrocytes owing to an imbalance between cell death and cell proliferation in the ACL insertions after rupture, as compared with normal insertions, may lead to histological changes of the cartilage layer in the insertions. An in-depth understanding of injured ACL insertion may help elucidate the etiology of histological changes and the function and significance of the existence of the cartilage layer of insertion. This understanding may help in developing optimal treatment protocols for ACL injuries if apoptosis and cell proliferation are controlled.     

小鼠出生后肾脏发育过程中的细胞增殖与凋亡

目的：观察小鼠出生后肾脏发育过程中增殖细胞核抗原(PCNA)的表达及细胞凋亡的特征,探讨出生后小鼠肾脏发育过程中细胞增殖与凋亡的规律及其关系。方法：应用免疫组织化学技术和原位末端标记法(TUNEL法)分别检测小鼠出生后1～70d肾脏中PCNA阳性的细胞和凋亡细胞。结果：小鼠出生后1～70d,皮质中的肾小体、肾小管、髓放线以及髓质中的肾小管和集合管的细胞,早期增殖活跃,随着肾脏发育成熟而表达逐渐减弱。同时,也存在着细胞凋亡现象,且凋亡高峰一般出现在增殖高峰之后。结论：细胞增殖与凋亡在小鼠生后肾脏发育的整个过程中普遍存在,生后1～7d细胞增殖旺盛,增殖高峰之后出现凋亡高峰,生后28～70d两者活动均减弱。     

Cell death and cell proliferation during atrophy of the rat parotid gland induced by duct obstruction

Rat parotid gland atrophy after unilateral duct ligation was studied by light and electron microscopy. Death of secretory acinar cells, which took the form of apoptosis, resulted in their complete disappearance within 5 days. The remnants of the dying cells were mostly phagocytosed and degraded by macrophages within the glandular epithelium; a few were taken up by adjoining epithelial cells. The acinar cell deletion was accompanied by increased mitosis of striated and intercalated duct epithelial cells. However, over many weeks, there was enhanced apoptosis of duct cells, which eventually led to marked shortening of intercalated ducts. Apoptosis of capillary endothelial cells was observed and may account for the reduction in the capillary bed known to accompany gland atrophy. The end-stage lesion comprised small numbers of ducts in a condensed stroma. Compensatory hyperplasia, involving proliferation of duct and acinar cells, was demonstrated in the contralateral glands.     

Apoptosis and proliferation in gastric carcinoma: the association with histological type

We examined apoptosis in 33 gastric carcinomas using the terminal deoxynucleotydil transferase mediated dUTP-digoxigenin nick end labelling technique (TUNEL). Of the tumours, nine were well-differentiated, 13 moderately differentiated and 11 poorly differentiated. In addition, we also analysed MIB-1, a cell proliferation antigen. Morphologically, apoptotic tumour cells were more frequently observed in well-differentiated tumours. In addition, apoptotic signals of the TUNEL method were seen even in the nuclei of tumour cells which did not show apoptosis. The nick end labelling index was 51.0 ± 26.3 in the well-differentiated and moderately differentiated tumours and 28.0 ± 18.8 in poorly differentiated tumours. The mean of apoptotic body index and nick end labelling index were both significantly higher in well-differentiated and moderately differentiated tumours than in the poorly differentiated type ( P  < 0.0001, P  = 0.008). The MIB-1 labelling index and higher in poorly differentiated tumours than in the well-differentiated or moderately differentiated tumours, and labelled cells were more numerous in the superficial region than in the middle and deep regions of tumours. No apparent correlation was found between the nick end labelling index and the MIB-1 labelling index. The high number of apoptotic cells (the high Nick end labelling index) and low proliferation potentiality (the low MIB-1 labelling index) in well-differentiated gastric carcinomas may thus be related to their natural tendency to demonstrate slow growth.     

Cell therapy of brain stroke

Cell suspension consisting of cells from immature nervous and hemopoietic tissues was subarachnoidally transplanted to 10 patients with brain stroke consequences. Clinical effect of different degree was attained in all patients. Six months after cell therapy functional activity significantly increased in contrast to clinically comparable control group. No serious complications of cell therapy were observed. Presumably, cell therapy is a more or less safe method of treatment, which can be effectively used in the treatment of brain stroke consequences.__________This revised version was published online in July 2005 with the addition of the issue title and article categoryTranslated from Kletochnye Tekhnologii v Biologii i Meditsine, Vol. 1, No. 1, pp. 25–28, January, 2005     

盐酸小檗碱对小鼠脾细胞增殖和凋亡及分泌细胞因子的影响

目的 研究盐酸小檗碱(BBR)对小鼠脾细胞增殖与凋亡及细胞因子产生的影响.方法 采用无菌取BALB/c小鼠脾脏,制成脾细胞悬液,脾细胞预先用(1、2、4) μg/mL BBR处理60 min,加入刀豆蛋白(ConA)刺激脾细胞增殖,细胞培养至24、48、72 h时,用MTT方法检测脾细胞增殖情况;流式细胞术检测脾细胞培养不同时间的凋亡情况,用细胞因子ELISA检测试剂盒定量分析细胞上清液中TNF-α,IL-2和IFN-γ的浓度.结果 与对照组相比,在以上浓度范围内,BBR对ConA刺激下实验组小鼠脾细胞的增殖和TNF-α,IL-2,IFN-γ产生有明显的抑制作用(P＜0.05),且呈浓度及时间依赖性,但对小鼠脾细胞凋亡无明显影响(P>0.05).结论 BBR对小鼠脾细胞具有明显的免疫抑制作用,可作为潜在的免疫抑制药物.     

Distribution of calretinin-like immunoreactivity in the brain of Rana esculenta.

The distribution of calretinin-like immunoreactivity has been analyzed in the brain of Rana esculenta. Several neurons of nuclei belonging to sensory pathways, subhabenular area and left habenula were immunopositive. Immunoreactivity was present in fibers of motor and sensory pathways, thalamus, tegmentum and isthmus. The immunolabeling pattern partially overlapped that previously described in the rat. However, in comparison with the rat, fewer cells and fibers were immunoreactive and there were less positive brain nuclei, especially in the pallium, septum and striatum, that were totally negative. Taking into consideration that these regions are rather simple in the frog, the presence of calretinin seems to be consistent with the degree of complexity of brain areas and segregation of different nuclei.     

Effects of an imprinting procedure on cell proliferation in the chick brain

We report here studies on the effects of an imprinting procedure on cell proliferation in neonatal chicks in brain structures known to undergo plastic changes in imprinting. Proliferating cells were detected immunohistochemically on brain sections by incorporation of pre-training doses of 5-bromodeoxyuridine (BrdU) into DNA; numbers of new cells were counted in the intermediate medial mesopallium, the intermediate arcopallium, the medial part of the mesopallium and the nidopallium, the dorsocaudal nidopallium, the hippocampus, and the parahippocampal region 24 h and seven days after training. The intermediate medial mesopallium showed an increase in the number of BrdU-positive cells 24 h after training. However, at seven days post-training, the number of BrdU-containing cells decreased in the medial nidopallium and mesopallium, in the dorsocaudal nidopallium, and the right intermediate medial mesopallium. Thus, the imprinting procedure had differently directed transient and long-term influences on the genesis of new cells in the chick brain, inducing the appearance of a large number of cells in the parenchyma of the brain one day after training and decreases in the numbers of cells at later time points. This double effect may be associated with the fact that the imprinting procedure simultaneously initiates two brain processes involving the control of cell proliferation — one related to maturation of a species-specific functional system for tracking individuals of the same species and one related to remembering the characteristics of the actual parent. __________ Translated from Zhurnal Vysshei Nervnoi Deyatel’nosti imeni I. P. Pavlova, Vol. 57, No. 2, pp. 181–190, March–April, 2007.     

应用流式细胞术分析不同年龄大鼠脑组织细胞周期增殖特性

应用流式细胞术对不同年龄的大鼠的大脑、小脑和脑干组织的细胞周期进行分析。结果显示在相同年龄组不同部位的脑组织细胞增殖程度不均一,大脑细胞增殖明显高于小脑和脑干。同一部位的脑组织各年龄组之间细胞增殖情况亦存在差异。新生和幼年大鼠(出生后1天和21天)其脑细胞增殖较成年大鼠(6个月和1年)旺盛。实验还表明,特殊萤光素染色脑细胞DNA,然后用流式细胞仪测定DNA量,是一种简单、快速分析脑细胞增殖活性的可行技术。     

Distribution of calretinin-like immunoreactivity in the brain of Rana esculenta

The distribution of calretinin-like immunoreactivity has been analyzed in the brain of Rana esculenta. Several neurons of nuclei belonging to sensory pathways, subhabenular area and left habenula were immunopositive. Immunoreactivity was present in fibers of motor and sensory pathways, thalamus, tegmentum and isthmus. The immunolabeling pattern partially overlapped that previously described in the rat. However, in comparison with the rat, fewer cells and fibers were immunoreactive and there were less positive brain nuclei, especially in the pallium, septum and striatum, that were totally negative. Taking into consideration that these regions are rather simple in the frog, the presence of calretinin seems to be consistent with the degree of complexity of brain areas and segregation of different nuclei.     

Cell cycle alterations and their relationship to proliferation in apocrine adenosis of the breast

Aims:  Apocrine adenosis (AA) is generally considered a benign disease of the breast. However, recent studies have suggested a precancerous potential for some of these lesions. The aim was to investigate the status of cell cycle proteins previously reported to be deregulated in breast cancer to identify their possible role in AA.
Methods and results:  The cases were categorized into AA without atypia (NAA) and atypical AA (AAA). Using immunohistochemistry, the expression of cyclin D1, cyclin A, p27, p21, p16, pRb and Ki67 was determined in 29 NAA and 16 AAA cases. Cyclin D1, p21 and cyclin A were overexpressed in 58.6%, 51.7% and 31.8% of the NAA cases, respectively, whereas 81.3%, 62.5% and 41.7% of the AAA cases showed overexpression of cyclin D1, p21 and cyclin A, respectively. All cases were negative for p16, whereas pRb was expressed in all cases. Furthermore, proliferation in AA (4.5%) was significantly higher than that of normal breast epithelium (1%). There was no statistical significance in the degree of proliferation between the NAA (3.7%) and AAA (4.8%) groups.
Conclusions:  The study indicates that NAA and AAA are biologically similar. A subset of AA defined by increased proliferation and significant cell cycle alterations may be susceptible to oncogenesis.     

大气压冷等离子体对肿瘤细胞增殖和凋亡的影响及其机制

目的:探讨大气压冷等离子体（CAP）对人肿瘤细胞增殖和凋亡的影响。方法:将处于对数生长期的人神经胶质瘤细胞（U87）和宫颈癌细胞（Hela）接种于培养板或培养皿中，待细胞贴壁生长后，将其随机分为正常对照（control）组、氦气（He）组和CAP不同时间处理组，细胞在接受不同时间CAP处理后，采用CCK-8方法检测细胞活力，流式细胞仪检测细胞凋亡情况，蛋白印迹法检测凋亡相关蛋白caspase3和氧化应激相关蛋白Nrf2的表达水平。结果:He处理对U87和Hela细胞的增殖能力和自发凋亡无明显影响，CAP处理后两种肿瘤细胞的细胞活力均显著降低，凋亡水平明显上升，caspase3蛋白和Nrf2蛋白的表达显著增加，CAP处理时间越长，效应越明显。结论:本实验条件下的CAP处理可抑制人肿瘤细胞的增殖，并诱导其凋亡，其机制可能与CAP导致的氧化应激有关。     

Role of neurosteroids in regulating cell death and proliferation in the late gestation fetal brain

The neurosteroid allopregnanolone (AP) is a GABAergic agonist that suppresses central nervous system (CNS) activity in the adult brain, and by reducing excitotoxicity is considered to be neuroprotective. A role for neurosteroids in the developing brain, particularly in late gestation, is still debated. The aim of this study was to investigate effects on proliferation and cell death in the brain of late gestation fetal sheep after inhibition of AP synthesis using finasteride, a 5α-reductase type 2 (5α-R2) inhibitor. Catheters were implanted in fetal sheep at ∼125 days of gestation. At 3–4 days postsurgery, fetuses received infusions of either finasteride (20 mg/kg/h; n=5), the AP analogue alfaxalone (5 mg/kg/h; n=5), or finasteride and alfaxalone together (n=5). Brains were obtained at 24 h after infusion to determine cell death (apoptotic or necrotic) and cell proliferation in the hippocampus and cerebellum, areas known to be susceptible to excitotoxic damage. Finasteride treatment significantly increased apoptosis (activated caspase-3 expression) in hippocampal CA3 and CA1, and cerebellar molecular and granular layers, an effect abolished by co-infusion of alfaxalone and finasteride. Double-label immunohistochemistry showed that both neurons and astrocytes were caspase-3 positive. Finasteride treatment also increased the number of dead (pyknotic) cells in the hippocampus and cerebellum (Purkinje cells), but not when finasteride+alfaxalone was infused. Cell proliferation (Ki-67-immunoreactivity) increased after finasteride treatment; double-labeling showed the majority of Ki-67-positive cells were astrocytes. Thus, steroids such as AP appear to influence the constitutive rate of apoptosis and proliferation in the hippocampus and cerebellum of the fetal brain, and suggest an important role for neurosteroids in the development of the brain.     

Monoaminc and metabolite levels in the cerebrospinal fluid of hibernating and euthermic marmots

SUMMARY  Cerebrospinal fluid from yellow-bellied marmots, Marmota flaviventris , was analysed for monoamine and monoamine metabolite content during euthermia and deep hibernation. Dopamine (DA) levels were decreased, while DA metabolite levels, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), were dramatically increased in hibernating marmots. Serotonin (5-HT) and 5-hydroxyindoleacetic acid (5HIAA) levels were also greatly enhanced during hibernation while norepinephrine (NE) levels were only moderately increased. These findings demonstrate that cerebrospinal monoamine levels are dynamically altered during hibernation, such that DA versus 5-HT and NE levels undergo opposite changes. Therefore, these data indicate that DA, 5-HT and NE neuronal systems are differentially altered during hibernation in mammals.     

谷胱甘肽对染锰大鼠生精细胞凋亡与增殖的影响

目的:研究谷胱甘肽(GSH)对染锰大鼠生精细胞凋亡与增殖的作用及机制。方法:雄性SD大鼠48只随机分为6组,各组给锰途径为腹腔注射。采用TUNEL法检测生精细胞凋亡指数(AI);免疫组织化学法检测生精细胞增殖细胞核抗原(PCNA)表达。结果:与空白对照组比较,GSH对照组和15mg/kg GSH组生精细胞AI与生精细胞PCNA增殖指数(PI)均无显著性差异;与各自对应的单纯染锰组比较,15mg/kg GSH组和30mg/kg GSH组生精细胞AI均显著降低而PI均显著升高;30mg/kg组与15mg/kg组比较,生精细胞AI显著升高而PI显著降低;各组大鼠生精细胞AI和PI呈负相关。结论:15mg/kg和30mg/kg氯化锰可诱发大鼠睾丸生精细胞凋亡,两者存在剂量-效应关系;各组大鼠睾丸生精细胞AI和PI呈显著负相关;GSH对锰诱发大鼠生精细胞凋亡具有拮抗作用;GSH可能拮抗锰对大鼠生精细胞增殖的抑制作用。