BACKGROUND & AIMS: :Several cAMP-elevating agents such as cholera toxin (CT), forskolin and 3-isobutyl-1-methylxanthine (IBMX) exhibited weak mitogenic activity on bovine undifferentiated mammary epithelial cells in three-dimensional collagen culture. CT and IBMX strongly synergized with epidermal growth factor (EGF), insulin-like growth factor I (IGF-I) or both, but not with 10% fetal calf serum (FCS). Permeable cAMP analogs also synergized with IGF-I. Other hormones such as ovine prolactin, bovine growth hormone, estrogen or progesterone were not mitogenic and not synergistic with EGF, IGF-I, CT and FCS. Pertussis toxin (PT) reduced the DNA synthesis in cells cultured in the basal medium and attenuated 40-90% of the mitogenic activity stimulated by 10% FCS. PT inhibition of DNA synthesis was accompanied by ADP-ribosylation of 40 kDa and 41 kDa membrane proteins. The 41 kDa protein cross-reacted with antibodies that recognize the Gi-protein of the adenylate cyclase system, indicating the involvement of the latter in the mitogenic process. The nature of the second protein remains unknown. The present results suggest that the mitogenesis of normal mammary epithelial cells which is stimulated by IGF-I, EGF and other factors found in FCS is mediated through both cAMP-dependent and independent pathways. These pathways include PT-sensitive GTP-binding proteins.

背景与目标： : 几种cAMP升高剂，例如霍乱毒素 (CT)，福司可林和3-异丁基-1-甲基黄嘌呤 (IBMX)，在三维胶原蛋白培养中对牛未分化的乳腺上皮细胞表现出弱的促有丝分裂活性。CT和IBMX与表皮生长因子 (EGF)，胰岛素样生长因子I (igf-i) 或两者强烈协同，但与10% 胎牛血清 (FCS) 不协同。可渗透的cAMP类似物也与igf-i协同作用。其他激素 (例如绵羊催乳素，牛生长激素，雌激素或孕激素) 没有促有丝分裂作用，并且与EGF，igf-i，CT和FCS没有协同作用。百日咳毒素 (PT) 减少了在基础培养基中培养的细胞中的DNA合成，并减弱了40-90% 由10% FCS刺激的促有丝分裂活性。PT抑制DNA合成伴随着40 kDa和41 kDa膜蛋白的ADP-核糖基化。41 kDa蛋白与识别腺苷酸环化酶系统的Gi蛋白的抗体发生交叉反应，表明后者参与了促有丝分裂过程。第二种蛋白质的性质仍然未知。目前的结果表明，FCS中发现的igf-i，EGF和其他因素刺激的正常乳腺上皮细胞的有丝分裂是通过cAMP依赖性和独立途径介导的。这些途径包括PT敏感的GTP结合蛋白。

BACKGROUND & AIMS: :Granulocyte colony-stimulating factor (GCSF) administration has been linked to the development of monosomy 7 in severe congenital neutropenia and aplastic anemia. We assessed the effect of pharmacologic doses of GCSF on monosomy 7 cells to determine whether this chromosomal abnormality developed de novo or arose as a result of favored expansion of a preexisting clone. Fluorescence in situ hybridization (FISH) of chromosome 7 was used to identify small populations of aneuploid cells. When bone marrow mononuclear cells from patients with monosomy 7 were cultured with 400 ng/ml GCSF, all samples showed significant increases in the proportion of monosomy 7 cells. In contrast, bone marrow from karyotypically normal aplastic anemia, myelodysplastic syndrome, or healthy individuals did not show an increase in monosomy 7 cells in culture. In bone marrow CD34 cells of patients with myelodysplastic syndrome and monosomy 7, GCSF receptor (GCSFR) protein was increased. Although no mutation was found in genomic GCSFR DNA, CD34 cells showed increased expression of the GCSFR class IV mRNA isoform, which is defective in signaling cellular differentiation. GCSFR signal transduction via the Jak/Stat system was abnormal in monosomy 7 CD34 cells, with increased phosphorylated signal transducer and activation of transcription protein, STAT1-P, and increased STAT5-P relative to STAT3-P. Our results suggest that pharmacologic doses of GCSF increase the proportion of preexisting monosomy 7 cells. The abnormal response of monosomy 7 cells to GCSF would be explained by the expansion of undifferentiated monosomy 7 clones expressing the class IV GCSFR, which is defective in signaling cell maturation.

背景与目标： : 粒细胞集落刺激因子 (GCSF) 的施用与严重先天性中性粒细胞减少症和再生障碍性贫血中7号单体的发展有关。我们评估了GCSF的药理剂量对7号单体细胞的影响，以确定这种染色体异常是从头出现还是由于预先存在的克隆的有利扩增而出现的。7号染色体的荧光原位杂交 (FISH) 用于鉴定非整倍体细胞的小群体。当用400 ng/ml GCSF培养来自7号单体患者的骨髓单个核细胞时，所有样品均显示7号单体细胞的比例显着增加。相反，来自核型正常再生障碍性贫血，骨髓增生异常综合征或健康个体的骨髓在培养物中未显示出7号单体细胞的增加。在骨髓增生异常综合征和7号单体患者的骨髓CD34细胞中，GCSF受体 (GCSFR) 蛋白升高。尽管在基因组GCSFR DNA中未发现突变，但CD34细胞显示GCSFR IV类mRNA同工型的表达增加，这在信号细胞分化方面存在缺陷。通过Jak/Stat系统的GCSFR信号转导在7号单体CD34细胞中异常，磷酸化信号转导增加，转录蛋白激活，STAT1-P，相对于STAT3-P，STAT5-P增加。我们的结果表明，GCSF的药理剂量增加了先前存在的7号单体细胞的比例。7号单体细胞对GCSF的异常反应可以通过表达IV类GCSFR的未分化的7号单体克隆的扩增来解释，后者在信号细胞成熟方面存在缺陷。

BACKGROUND & AIMS: :B7-H3 is an immune regulatory molecule whose aberrant expression in tumors is associated with adverse outcomes. Upregulation of B7-H3 may promote tumor cell proliferation and metastasis in vitro, but the role of B7-H3 in cervical cancer has not yet been investigated. We measured B7-H3 expression in 90 cervical cancer patient and 20 non‑cervical lesion patient tissues using immunohistochemistry and in 30 cervical cancer patient and 30 healthy donor blood samples using ELISA. The association of B7-H3 expression and the prognosis of cervical cancer patients was investigated. B7-H3 knockdown in CaSki and SiHa cell lines was performed using small hairpin (sh)RNA lentiviral transfection and B7-H3 overexpression in CaSki and HeLa cell lines was performed using plasmid-vector lentivirus transduction. Cell proliferation, invasion and migration were then measured using MTT and Transwell assays in vitro. B7-H3 expression was significantly higher in the cervical cancer tissues compared to that noted in the normal cervical tissues (mean 72.22 vs. 15.00%; p<0.001). Using Kaplan‑Meier and Cox analyses, our data revealed that patients with strong intensity staining were significantly more likely to have a worse prognosis. The B7-H3 level in cervical cancer patient blood was significantly higher than that in the normal donors (13.41±6.12 vs. 9.90±3.16 ng/ml; p=0.007). MTT assay revealed that high expression of B7-H3 promoted cervical cancer cell proliferation. Transwell assay data revealed that high expression of B7-H3 enhanced cervical cancer cell migration and invasion (CaSki, p=0.003; HeLa, p=0.03). In conclusion, expression of B7-H3 was significantly higher in cervical cancer tissues compared to normal cervical tissues, and this high expression was associated with worse prognosis for cervical cancer patients. In addition, B7-H3 promoted proliferation, invasion and migration of cervical cancer and may be a potential target for treating cervical cancer.

背景与目标： : B7-H3是一种免疫调节分子，其在肿瘤中的异常表达与不良后果有关。B7-H3的上调可能在体外促进肿瘤细胞的增殖和转移，但B7-H3在宫颈癌中的作用尚未得到研究。我们使用免疫组织化学方法测量了90例宫颈癌患者和20例非宫颈病变患者组织中的B7-H3表达，并使用ELISA测量了30例宫颈癌患者和30例健康供体血样中的表达。研究了B7-H3表达与宫颈癌患者预后的关系。使用小发夹 (sh)RNA慢病毒转染在CaSki和SiHa细胞系中进行B7-H3敲除，并使用质粒载体慢病毒转导在CaSki和HeLa细胞系中进行B7-H3过表达。细胞增殖，然后在体外使用MTT和Transwell测定法测量侵袭和迁移。与正常宫颈组织相比，宫颈癌组织中的B7-H3表达明显更高 (平均72.22对15.00%; p<0.001)。使用kaplan-meier和Cox分析，我们的数据显示，强染色的患者预后更差。宫颈癌患者血液中的B7-H3水平明显高于正常供体 (13.41 ± 6.12 vs. 9.90 ± 3.16 ng/ml); p = 0.007)。MTT分析显示B7-H3的高表达促进了宫颈癌细胞的增殖。tranwell分析数据显示B7-H3的高表达增强了宫颈癌细胞的迁移和侵袭 (CaSki，p = 0.003; HeLa，p = 0.03)。总之，b7-H3在宫颈癌组织中的表达明显高于正常宫颈组织，这种高表达与宫颈癌患者预后差有关，此外，B7-H3促进了宫颈癌的增殖、侵袭和迁移，可能是治疗宫颈癌的潜在靶点。

BACKGROUND & AIMS: :Electrical fields are known to interact with human cells. This principle has been explored to regulate cellular activities for bone tissue regeneration. In this work, Saos-2 cells were cultured on conductive scaffolds made of biodegradable poly(L-lactide) and the heparin-containing, electrically conducting polypyrrole (PPy/HE) to study their reaction to electrical stimulation (ES) mediated through such scaffolds. Both the duration and intensity of ES enhanced cell proliferation, generating a unique electrical intensity and temporal "window" within which osteoblast proliferation was upmodulated in contrast to the downmodulation or ineffectiveness in other ES regions. The favourable ES intensity (200 mV/mm) was further investigated in terms of the gene activation and protein production of two important osteoblast markers characterised by extracellular matrix maturation and mineralisation, that is alkaline phosphatase (ALP) and osteocalcin (OC). Both genes were found activated and the relevant protein production increased significantly following ES. In contrast, ES in the down-modulation region (400 mV/mm) suppressed the production of both ALP and OC. This work demonstrated that important osteoblast markers can be modulated with specific ES parameters mediated through conductive polymer substrates, providing a unique strategy for bone tissue engineering.

背景与目标： 已知电场与人类细胞相互作用。已探索此原理来调节骨组织再生的细胞活性。在这项工作中，将Saos-2细胞培养在由可生物降解的聚 (L-丙交酯) 和含肝素的导电聚吡咯 (PPy/HE) 制成的导电支架上，以研究它们对通过此类支架介导的电刺激 (ES) 的反应。ES的持续时间和强度都增强了细胞增殖，产生了独特的电强度和时间 “窗口”，与其他ES区域的下调或无效相反，在该窗口内对成骨细胞增殖进行了上调。进一步研究了有利的ES强度 (200  mV/mm)，即两种重要的成骨细胞标志物的基因活化和蛋白质生产，其特征是细胞外基质成熟和矿化，即碱性磷酸酶 (ALP) 和骨钙素 (OC)。发现两个基因都被激活，并且在ES之后相关的蛋白质产量显着增加。相反，下调制区域 (400  mV/mm) 中的ES抑制了ALP和OC的产生。这项工作表明，重要的成骨细胞标志物可以通过导电聚合物底物介导的特定ES参数进行调节，为骨组织工程提供了独特的策略。

BACKGROUND & AIMS: :After nerve injury, Schwann cells proliferate and revert to a phenotype that supports nerve regeneration. This phenotype-changing process can be viewed as Schwann cell dedifferentiation. Here, we investigated the role of extracellular ATP in Schwann cell dedifferentiation and proliferation during Wallerian degeneration. Using several markers of Schwann cell dedifferentiation and proliferation in sciatic explants, we found that extracellular ATP inhibits Schwann cell dedifferentiation and proliferation during Wallerian degeneration. Furthermore, the blockage of lysosomal exocytosis in ATP-treated sciatic explants is sufficient to induce Schwann cell dedifferentiation. Together, these findings suggest that ATP-induced lysosomal exocytosis may be involved in Schwann cell dedifferentiation.

背景与目标： : 神经损伤后，雪旺氏细胞增殖并恢复为支持神经再生的表型。这种表型变化过程可以看作是雪旺细胞去分化。在这里，我们研究了细胞外ATP在华勒变性过程中雪旺细胞去分化和增殖中的作用。使用坐骨外植体中雪旺细胞去分化和增殖的几种标记，我们发现细胞外ATP在华勒变性过程中抑制雪旺细胞去分化和增殖。此外，ATP处理的坐骨外植体中溶酶体胞吐作用的阻断足以诱导雪旺氏细胞去分化。总之，这些发现表明ATP诱导的溶酶体胞吐作用可能与雪旺氏细胞去分化有关。

BACKGROUND & AIMS: AIM:To investigate the expression patterns of TTYH2 in the human colon cancer and colon cancer cell lines and to evaluate the inhibitory effect of small interfering RNA (siRNA) on the expression of TTYH2 in colon cancer cell lines. METHODS:We investigated the expression patterns of TTYH2 in colon cancer, adjacent non-tumorous colon mucosa, and cancer cell lines (DLD-1, caco-2, and Lovo) by RT-PCR. Furthermore, a siRNA plasmid expression vector against TTYH2 was constructed and transfected into DLD-1 and Caco-2 with Lipofectamine 2000. The down regulation of TTYH2 expression was detected by RT-PCR and the role of siRNA in inducing cell proliferation and cell aggregation was evaluated by MTT and aggregation assay. RESULTS:TTYH2 gene expression in colon cancer tissue was significantly up-regulated compared with normal colonic mucosa (1.23 +/- 0.404 vs 0.655 +/- 0.373, P = 0.0103). Colon cancer derived cell lines including DLD-1, Caco-2, and Lovo also expressed high levels of TTYH2. In contrast, transfection with siRNA-TTYH2 significantly inhibited both proliferation and scattering of these cancer cell lines. CONCLUSION:The present work demonstrates, for the first time, that the TTYH2 gene expression is significantly up-regulated in colon cancer. The TTYH2 gene may play an important role in regulating both proliferating and metastatic potentials of colorectal cancer.

背景与目标：

BACKGROUND & AIMS: :MicroRNAs (miRs), a class of non-coding RNAs that are 18‑25 nucleotides in length, serve as key regulators in the development and progression of human cancers. Previously, miR‑503 has been implicated in breast cancer. However, the underlying mechanism of miR‑503 in regulating the proliferation and invasion of breast cancer cells remains largely unknown. In the present study, reverse transcription‑quantitative polymerase chain reaction analysis indicated that the expression of miR‑503 was significantly reduced in breast cancer tissues compared with their matched adjacent normal tissues. Furthermore, miR‑503 expression levels were markedly reduced in T2‑T4 stage breast cancer, compared with T1 stage. Insulin‑like growth factor 1 receptor (IGF‑1R) was further identified as a novel target of miR‑503. Overexpression of miR‑503 significantly suppressed the protein expression levels of IGF‑1R. Furthermore, it inhibited the proliferation and invasion of human breast cancer MCF‑7 cells, as assessed by MTT and Transwell assays, respectively. However, restoration of IGF‑1R expression markedly ameliorated the suppressive effects of miR‑503 overexpression on MCF‑7 cell proliferation and invasion, indicating that miR‑503 inhibits breast cancer cell proliferation and invasion at least partially via directly targeting IGF‑1R. Furthermore, the mRNA and protein expression levels of IGF‑1R were demonstrated to be significantly increased in breast cancer tissues compared with their matched adjacent normal tissues. In addition, IGF‑1R mRNA expression levels were reversely correlated with miR‑503 expression levels in breast tumors, suggesting that the upregulation of IGF‑1R may be due to downregulation of miR‑503 in breast cancer. In conclusion, the present study expanded the understanding of the regulatory mechanism of miR‑503 in breast cancer, and implicates the miR‑503/IGF‑1R axis as a potential therapeutic target for breast cancer.

背景与目标： : microrna (miRs) 是一类长度为18-25个核苷酸的非编码rna，是人类癌症发生和发展的关键调节剂。以前，mir-503与乳腺癌有关。然而，mir-503在调节乳腺癌细胞增殖和侵袭中的潜在机制在很大程度上仍然未知。在本研究中，逆转录定量聚合酶链反应分析表明，与匹配的邻近正常组织相比，乳腺癌组织中mir-503的表达显着降低。此外，与T1期相比，T2-T4期乳腺癌的mir-503表达水平显著降低。胰岛素样生长因子1受体 (igf ‑ 1R) 被进一步鉴定为mir-503的新靶标。Mir-503的过表达显著抑制igf ‑ 1R的蛋白表达水平。此外，它抑制人乳腺癌mcf ‑ 7细胞的增殖和侵袭，分别通过MTT和tranwell分析评估。然而，igf ‑ 1R表达的恢复显著改善了mir-503过表达对mcf ‑ 7细胞增殖和侵袭的抑制作用，表明mir-503至少部分通过直接靶向igf ‑ 1R来抑制乳腺癌细胞增殖和侵袭。此外，与匹配的邻近正常组织相比，乳腺癌组织中igf ‑ 1R的mRNA和蛋白表达水平显着增加。此外，igf ‑ 1R mRNA表达水平与乳腺肿瘤中的503表达水平呈反向相关，这表明igf ‑ 1R的上调可能是由于乳腺癌中mir-503的下调所致。总之，本研究扩大了对乳腺癌中mir-503调控机制的理解，并暗示mir-503/igf ‑ 1R轴是乳腺癌的潜在治疗靶标。

BACKGROUND & AIMS: :Although the involvement of Rho proteins in the pathogenesis of vascular diseases is well studied, little is known about the role of their upstream regulators, the Rho guanine nucleotide exchange factors (RhoGEFs). Here, we sought to identify the RhoGEFs involved in monocyte chemotactic protein 1 (MCP1)-induced vascular wall remodeling. We found that, among the RhoGEFs tested, MCP1 induced tyrosine phosphorylation of p115 RhoGEF but not of PDZ RhoGEF or leukemia-associated RhoGEF in human aortic smooth muscle cells (HASMCs). Moreover, p115 RhoGEF inhibition suppressed MCP1-induced HASMC migration and proliferation. Consistent with these observations, balloon injury (BI) induced p115 RhoGEF tyrosine phosphorylation in rat common carotid arteries, and siRNA-mediated down-regulation of its levels substantially attenuated BI-induced smooth muscle cell migration and proliferation, resulting in reduced neointima formation. Furthermore, depletion of p115 RhoGEF levels also abrogated MCP1- or BI-induced Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1 signaling, which, as we reported previously, is involved in vascular wall remodeling. Our findings also show that protein kinase N1 (PKN1) downstream of Rac1-cyclin D1/CDK6 and upstream of CDK4-PAK1 in the p115 RhoGEF-Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1 signaling axis is involved in the modulation of vascular wall remodeling. Of note, we also observed that CCR2-Gi/o-Fyn signaling mediates MCP1-induced p115 RhoGEF and Rac1 GTPase activation. These findings suggest that p115 RhoGEF is critical for MCP1-induced HASMC migration and proliferation in vitro and for injury-induced neointima formation in vivo by modulating Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1 signaling.

背景与目标： : 尽管对Rho蛋白参与血管疾病的发病机理进行了很好的研究，但对其上游调节剂Rho鸟嘌呤核苷酸交换因子 (rhoiefs) 的作用知之甚少。在这里，我们试图鉴定参与单核细胞趋化蛋白1 (MCP1) 诱导的血管壁重塑的RhoGEFs。我们发现，在测试的RhoGEFs中，MCP1诱导人主动脉平滑肌细胞 (HASMCs) 中p115 RhoGEF的酪氨酸磷酸化，但不诱导PDZ RhoGEF或与白血病相关的RhoGEF的酪氨酸磷酸化。此外，p115 rhoge抑制抑制了MCP1-induced HASMC的迁移和增殖。与这些观察结果一致，球囊损伤 (BI) 在大鼠颈总动脉中诱导了p115 RhoGEF酪氨酸磷酸化，而siRNA介导的其水平下调大大减弱了BI诱导的平滑肌细胞迁移和增殖，从而减少了新内膜的形成。此外，p115 RhoGEF水平的耗竭也消除了MCP1或BI诱导的Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1信号传导，正如我们先前报道的那样，这与血管壁重塑有关。我们的发现还表明，Rac1-cyclin D1/CDK6下游和p115 RhoGEF-Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1信号轴CDK4-PAK1上游的蛋白激酶N1 (PKN1) 参与了血管壁重塑的调节。值得注意的是，我们还观察到CCR2-Gi/o-Fyn信号介导MCP1-induced p115 rhoge和rac1gtpase激活。这些发现表明，p115 RhoGEF对于MCP1-induced HASMC在体外迁移和增殖以及通过调节Rac1-NFATc1-cyclin D1-CDK6-PKN1-CDK4-PAK1信号传导在体内损伤诱导的新内膜形成至关重要。

BACKGROUND & AIMS: OBJECTIVE:To investigate the antifibrotic role of rosmarinic acid (RA), a natural polyphenolic compound, on HSCs activation/proliferation and apoptosis in vitro and in vivo. METHODS:The impact of RA on stellate cell line (HSC-T6) proliferation, activation and apoptosis was assessed along with its safety on primary hepatocytes. In vivo, rats were divided into: (i) normal; (ii) thioacetamide (TAA)-intoxicated rats for 12 weeks; (iii) TAA + silymarin or (iv) TAA + RA. At the end of experiment, liver functions, oxidative stress, inflammatory and profibrogenic markers, tissue inhibitor metalloproteinases type-1 (TIMP-1) and hydroxyproline (HP) levels were evaluated. Additionally, liver histopathology and immunohistochemical examinations of alpha-smooth muscle actin (α-SMA), caspase-3 and proliferation cellular nuclear antigen (PCNA) were determined. RESULTS:RA exhibited anti-proliferative effects on cultured HSCs in a time and concentration dependent manner showing an IC50 of 276 μg/mL and 171 μg/mL for 24 h and 48 h, respectively, with morphological reversion of activated stellate cell morphology to quiescent form. It significantly improved ALT, AST, oxidative stress markers and reduced TIMP-1, HP levels, inflammatory markers and fibrosis score (S1 vs S4). Furthermore, reduction in α-SMA plus elevation in caspase-3 expressions of HSCs in vitro and in vivo associated with an inhibition in proliferation of damaged hepatocytes were recorded. CONCLUSIONS:RA impeded the progression of liver fibrosis through inhibition of HSCs activation/proliferation and induction of apoptosis with preservation of hepatic architecture.

背景与目标：

BACKGROUND & AIMS: :The diffusible messenger, nitric oxide plays multiple roles in neuroprotection, neurodegeneration and brain plasticity. Its involvement in neurogenesis has been disputed, on the basis of results on models in vivo and in culture. We report here that pharmacological blockade of nitric oxide production in rat pups resulted, during a restricted time window of the first three postnatal days, in increased cerebellar proliferation rate, as assessed through tritiated thymidine or BrdU incorporation into DNA. This was accompanied by increased expression of Myc, a transcription factor essential for cerebellar development, and of the cell cycle regulating gene, cyclin D1. These effects were mediated downstream by the nitric oxide-dependent second messenger, cGMP. Schedules of pharmacological NO deprivation targeted to later developmental stages (from postnatal day 3 to 7), no longer increased proliferation, probably because of partial escape of the cGMP level from nitric oxide control. Though limited to a brief temporal window, the proliferative effect of neonatal nitric oxide deprivation could be traced into adulthood. Indeed, the number of BrdU-labeled surviving cells, most of which were of neuronal phenotype, was larger in the cerebellum of 60-day-old rats that had been subjected to NO deprivation during the first three postnatal days than in control rats. Experiments on cell cultures from neonatal cerebellum confirmed that nitric oxide deprivation stimulated proliferation of cerebellar precursor cells and that this effect was not additive with the proliferative action of sonic hedgehog peptide. The finding that nitric oxide deprivation during early cerebellar neurogenesis, stimulates a brief increase in cell proliferation may contribute to a better understanding of the controversial role of nitric oxide in brain development.

背景与目标： 一氧化氮是可扩散的信使，在神经保护，神经变性和大脑可塑性中起着多种作用。根据体内和培养模型的结果，它在神经发生中的参与存在争议。我们在这里报告说，通过trisitation胸苷或BrdU掺入DNA来评估，在出生后的前三天的有限时间窗口内，大鼠幼崽中一氧化氮产生的药理学阻断导致小脑增殖率增加。伴随着Myc (小脑发育必不可少的转录因子) 和细胞周期调节基因cyclin d1的表达增加。这些作用是由一氧化氮依赖性第二信使cGMP介导的。针对后期发育阶段 (从出生后第3天到第7天) 的药理学无剥夺时间表，不再增加增殖，这可能是由于cGMP水平从一氧化氮控制中部分逸出所致。尽管仅限于短暂的颞窗，但新生儿一氧化氮剥夺的增殖作用可以追溯到成年。实际上，在60天大的大鼠的小脑中，BrdU标记的存活细胞的数量 (其中大多数是神经元表型) 在出生后的前三天没有遭受剥夺的大鼠比在对照组中更大。对新生儿小脑细胞培养物的实验证实，一氧化氮的剥夺刺激了小脑前体细胞的增殖，并且这种作用与sonic hedgehog肽的增殖作用无关。发现小脑早期神经发生过程中一氧化氮剥夺会刺激细胞增殖的短暂增加，这可能有助于更好地理解一氧化氮在大脑发育中的有争议作用。

BACKGROUND & AIMS: :Although many multiple myeloma (MM) patients initially respond to cytotoxic therapy, most eventually relapse. Novel therapeutic strategies employing a combination of chemotherapy with targeted biologics may significantly enhance the response of tumor cells to treatment. We tested a fully human anti-IGF-IR antibody (A12) against MM, and showed specific inhibition of IGF-I or serum-induced IGF-IR signaling in MM cells in vitro. The A12 as a single agent was demonstrated to exert modest to significant inhibition of tumor growth in vivo in various subcutaneous xenograft MM models. The A12 was also evaluated in a disseminated xenograft MM.1S NOD/SCID model as monotherapy or in combination with other drugs (bortezomib, melphalan) currently in clinical use. The tumor burden, as determined by luciferase bioimaging, was sharply decreased, and overall survival significantly prolonged when the therapies were combined. Immunohistochemical analysis demonstrated that the A12 treated tumors had significantly decreased vascularization compared to control tumors. Furthermore, most MM lines constitutively secreted significant quantities of VEGF, and this was enhanced following IGF-I treatment. Inhibition of IGF-IR by the A12 in vitro suppressed both constitutive and IGF-I-induced secretion of VEGF, indicating that a putative anti-angiogenic mechanism associated with the A12 treatment may contribute to its anti-tumor effect.

背景与目标： : 尽管许多多发性骨髓瘤 (MM) 患者最初对细胞毒性治疗有反应，但大多数最终复发。采用化疗与靶向生物制剂相结合的新型治疗策略可能会显着增强肿瘤细胞对治疗的反应。我们测试了针对MM的全人抗igf-ir抗体 (A12)，并在体外显示了对MM细胞中igf-i或血清诱导的igf-ir信号的特异性抑制。在各种皮下异种移植MM模型中，A12作为单一药物被证明对体内肿瘤生长具有适度至显着的抑制作用。还在播散性异种移植MM.1S NOD/SCID模型中评估了A12，作为单一疗法或与目前临床使用的其他药物 (硼替佐米，美法仑) 联合使用。通过荧光素酶生物成像确定的肿瘤负荷急剧降低，并且当联合治疗时，总生存期显着延长。免疫组织化学分析表明，与对照肿瘤相比，A12治疗的肿瘤血管形成明显减少。此外，大多数MM系组成型分泌了大量的VEGF，并且在igf-i治疗后这种情况得到了增强。体外A12抑制igf-ir抑制了组成型和igf-i诱导的VEGF分泌，表明与A12治疗相关的推定抗血管生成机制可能有助于其抗肿瘤作用。

BACKGROUND & AIMS: AIM:To investigate the effects of inositol hexaphosphate (IP(6)) on proliferation of HT-29 human colon carcinoma cell line. METHODS:Cells were exposed to various concentrations (0, 1.8, 3.3, 5.0, 8.0, 13.0 mmol/L) of IP(6) for a certain period of time. Its effect on growth of HT-29 cells was measured by MTT assay. The expressions of cell cycle regulators treated with IP(6) for 2 d were detected by immunocytochemistry. RESULTS:IP(6) inhibited the HT-29 cell growth in a dose- and time-dependent manner. Analysis of cell cycle regulator expression revealed that IP(6) reduced the abnormal expression of P53 and PCNA and induced the expression of P21. CONCLUSION:IP(6) has potent inhibitory effect on proliferation of HT-29 cells by modulating the expression of special cell cycle regulators.

背景与目标：

BACKGROUND & AIMS: :The individual roles of the two TNFRs on dendritic cells (DC) are poorly understood. Investigating bone marrow-derived DC from TNFR-deficient mice, we found that cultures from TNFR1(-/-) mice continue to form proliferating clusters for 6-9 mo. In contrast, DC derived from wild-type, TNFR2(-/-), or TNFR1/2(-/-) mice survived for only 3-4 wk. DC obtained from these TNFR1(-/-) long term cultures (LTC) mice show an unusual mixed immature/mature phenotype. The continuous proliferation of the LTC is GM-CSF dependent and correlates with decreased protein levels of the cyclin-dependent kinase inhibitors p27(KIP1) and p21(CIP1). Prolonged survival of TNFR1(-/-) DC appears to be independent from NF-kappaB and Bcl-2 pathways and is rather enabled by the down-regulation of CD95, resulting in the resistance to CD95 ligand-induced apoptosis. These data point to proapoptotic signals mediated via TNFR1 and antiapoptotic signals mediated via TNFR2 in DC.

背景与目标： : 两种tnfr对树突状细胞 (DC) 的个体作用知之甚少。研究来自TNFR缺陷型小鼠的骨髓来源的DC，我们发现来自TNFR1(-/-) 小鼠的培养物在6-9个月内继续形成增殖簇。相反，来自野生型，TNFR2(-/-) 或TNFR1/2(-/-) 小鼠的DC仅存活3-4周。从这些TNFR1(-/-) 长期培养 (LTC) 小鼠获得的DC显示出异常的混合未成熟/成熟表型。LTC的持续增殖是gm-csf依赖性的，并且与细胞周期蛋白依赖性激酶抑制剂p27(KIP1) 和p21(CIP1) 的蛋白水平降低相关。TNFR1(-/-) DC的长期存活似乎独立于NF-κ b和Bcl-2途径，并且由于CD95的下调而导致对CD95配体诱导的凋亡的抗性。这些数据指向DC中通过TNFR1介导的促凋亡信号和通过TNFR2介导的抗凋亡信号。

BACKGROUND & AIMS: :Nanomaterials hold great promise in the manipulation and treatments of mesenchymal stem cells, since they allow the modulation of their properties and differentiation. However, systematic studies have to be carried out in order to assess their potential toxicological effects. The present study reports on biocompatibility evaluation of glycol-chitosan coated barium titanate nanoparticles (BTNPs) on rat mesenchymal stem cells (MSCs). BTNPs are a class of ceramic systems which possess interesting features for biological applications thanks to their peculiar dielectric and piezoelectric properties. Viability was evaluated up to 5 days of incubation (concentrations in the range 0-100 μg/ml) both quantitatively and qualitatively with specific assays. Interactions cells/nanoparticles were further investigated with analysis of the cytoskeleton conformation, with SEM and TEM imaging, and with AFM analysis. Finally, differentiation in adipocytes and osteocytes was achieved in the presence of high doses of BTNPs, thus highlighting the safety of these nanostructures towards mesenchymal stem cells.

背景与目标： : 纳米材料在间充质干细胞的操作和治疗中具有广阔的前景，因为它们可以调节其特性和分化。但是，必须进行系统的研究以评估其潜在的毒理学作用。本研究报道了乙二醇-壳聚糖包被的钛酸钡纳米颗粒 (BTNPs) 对大鼠间充质干细胞 (MSCs) 的生物相容性评价。BTNPs是一类陶瓷系统，由于其独特的介电和压电特性，具有用于生物应用的有趣功能。通过特定测定定量和定性地评估孵育5天 (浓度在0-100 μ g/ml范围内) 的活力。通过分析细胞骨架构象，SEM和TEM成像以及AFM分析进一步研究了细胞/纳米颗粒的相互作用。最后，在高剂量BTNPs的存在下实现了脂肪细胞和骨细胞的分化，从而突出了这些纳米结构对间充质干细胞的安全性。

BACKGROUND & AIMS: :Aberrant metabolism is a hallmark of cancer, but whole metabolomic flux measurements remain scarce. To bridge this gap, we developed a novel metabolic phenotypic analysis (MPA) method that infers metabolic phenotypes based on the integration of transcriptomics or proteomics data within a human genome-scale metabolic model. MPA was applied to conduct the first genome-scale study of breast cancer metabolism based on the gene expression of a large cohort of clinical samples. The modeling correctly predicted cell lines' growth rates, tumor lipid levels, and amino acid biomarkers, outperforming extant metabolic modeling methods. Experimental validation was obtained in vitro. The analysis revealed a subtype-independent "go or grow" dichotomy in breast cancer, where proliferation rates decrease as tumors evolve metastatic capability. MPA also identified a stoichiometric tradeoff that links the observed reduction in proliferation rates to the growing need to detoxify reactive oxygen species. Finally, a fundamental stoichiometric tradeoff between serine and glutamine metabolism was found, presenting a novel hallmark of estrogen receptor (ER)(+) versus ER(-) tumor metabolism. Together, our findings greatly extend insights into core metabolic aberrations and their impact in breast cancer.

背景与目标： : 异常代谢是癌症的标志，但整个代谢组通量测量仍然很少。为了弥合这一差距，我们开发了一种新颖的代谢表型分析 (MPA) 方法，该方法基于人类基因组规模代谢模型中转录组学或蛋白质组学数据的整合来推断代谢表型。根据大量临床样本的基因表达，将MPA应用于乳腺癌代谢的首次基因组规模研究。该模型正确预测了细胞系的生长速率，肿瘤脂质水平和氨基酸生物标志物，优于现有的代谢建模方法。在体外获得了实验验证。分析揭示了乳腺癌中独立于亚型的 “去或生长” 二分法，随着肿瘤发展转移能力，增殖率降低。MPA还确定了化学计量的权衡，该权衡将观察到的增殖速率降低与对活性氧的解毒需求的增长联系起来。最后，发现丝氨酸和谷氨酰胺代谢之间的基本化学计量权衡，呈现了雌激素受体 (ER)() 与ER(-) 肿瘤代谢的新标志。总之，我们的发现极大地扩展了对核心代谢异常及其对乳腺癌影响的见解。