Fasudil mediates cell therapy of EAE by immunomodulating encephalomyelitic T cells and macrophages

Although Fasudil has shown therapeutic potential in EAE mice, the mechanism of action are still not fully understood. Here, we examined the immunomodulatory effect of Fasudil on encephalitogenic mononuclear cells (MNCs), and tested the therapeutic potential of Fasudil‐treated MNCs in active EAE. Fasudil inhibited expression of CCL20 on T cells and migration of T cells, decreased CD4⁺IFN‐γ⁺ and CD4⁺IL‐17⁺ T cells, but increased CD4⁺IL‐10⁺ and CD4⁺TGF‐β⁺ T cells. Fasudil reduced expression of CD16/32 and IL‐12, while elevating expression of CD206, CD23, and IL‐10. Fasudil also decreased levels of iNOS/NO, enhanced levels of Arg‐1, and inhibited the TLR‐4/NF‐κB signaling and TNF‐α, shifting M1 macrophage to M2 phenotype. These modulatory effects of Fasudil on T cells and macrophages were not altered by adding autoantigen MOG_35–55 to the culture, i.e., autoantigen‐independent. Further, we observed that, in vitro, Fasudil inhibited the capacity of encephalitogenic MNCs to adoptively transfer EAE and reduced TLR‐4/p‐NF‐κB/p65 and inflammatory cytokines in spinal cords. Importantly, Fasudil‐treated encephalitogenic MNCs exhibited therapeutic potential when injected into actively induced EAE mice. Together, our results not only provide evidence that Fasudil mediates the polarization of macrophages and the regulation of T cells, but also reveal a novel strategy for cell therapy in MS.     

Anti-epidermal growth factor receptor monoclonal antibody-based therapy for metastatic colorectal cancer: a meta-analysis of the effect of PIK3CA mutations in KRAS wild-type patients

Introduction

We conducted a meta-analysis to dissect the association between PIK3CA mutations (exon 9 and exon 20) and resistance to anti-epidermal growth factor receptor (EGFR) monoclonal antibodies (MoAbs) in KRAS wild-type metastatic colorectal cancer (mCRC) patients.

Material and methods

In 11 previously published studies, 864 cancer patients were treated with cetuximab or panitumumab-based therapy. Primary outcomes included objective response (complete response + partial response vs. stable disease + progressive disease), progression-free survival (PFS), and overall survival (OS). We calculated the odds ratio (OR) or hazard ratio (HR) with 95% confidence intervals (CIs) to estimate the risk or hazard. We found consistent and clinically substantial risk or hazard for objective response, PFS, and OS in the cetuximab or panitumumab-treated mCRC patients.

Results

PIK3CA mutations as a whole were associated with reduced response and poor PFS and OS in KRAS wild-type mCRC patients (objective response: OR = 0.42 and 95% CI 0.23–0.75; PFS: HR = 1.54 and 95% CI 1.13–2.09; and OS: HR = 1.4 and 95% CI 1.02–1.91). PIK3CA exon 9 mutations had no effect, whereas exon 20 mutations were associated with a worse outcome compared with wild types, with an OR of 0.21 (95% CI 0.05–0.93).

Conclusions

PIK3CA mutations as a whole might be useful prognostic factors for assessing clinical outcomes of anti-EGFR MoAb-based chemotherapies in KRAS wild-type mCRC patients. In particular, PIK3CA exon 20 mutations were significantly associated with lack of response.     

Effect of strontium ions on the growth of ROS17/2.8 cells on porous calcium polyphosphate scaffolds

Preparation, characterization and cellular biocompatibility study of a series of calcium polyphosphate containing 0-100 mol% of Ca2+ replaced by Sr2+ were reported. The osteoblastic ROS17/2.8 cell line was used and seeded on the strontium-doped calcium polyphosphate (SCPP) scaffolds to estimate its optimal dose and to study its potential to support the growth of osteoblastic cells for bone tissue engineering. The effects of SCPP on cells' proliferation and differentiation were evaluated by MTT and ALP activity assay. The results showed that porous SCPP did not exert cytotoxic effect on the cells. In addition, the proliferation and differentiation of the growth of ROS17/2.8 cells on the SCPP containing a low dose of strontium showed a higher level compared to the control, and the SCPP containing 1% strontium was optimal according to the results of MTT and ALP activity assay. The cells on the porous SCPP formed a continuous layer on the outer and inner surface observed by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The bunchy collagens were excreted from the cells and the calcium granules wrapped by collagens were sedimentated on the surface of cells. The results suggested that the biodegradable SCPP could stimulate the proliferation and differentiation of ROS17/2.8 cells in vitro after addition of proper dose of strontium. The porous SCPP may be a promising material for the bone tissue engineering.     

Abnormal Development at Early Postimplantation Stage in Mouse Embryos After Preimplantation Genetic Diagnosis

Preimplantation genetic diagnosis (PGD) is an established procedure for the genetic analysis of embryos. To assess the effect of the procedure on early embryonic development, we generated a murine experimental system, including mice implanted with biopsied in vitro cultured embryos, control mice implanted with in vitro cultured embryos without biopsy, and mice with naturally conceived embryos. Embryos at the 7.5‐dpc stage were isolated from all three groups and the embryo implantation rate, the survival rate of implanted embryos, and the developmental stage of surviving embryos were carefully assessed and compared among all three groups. We found the implantation rate was similar between biopsied and control group embryos (67.92% vs. 66.67%). However, the survival rate of implanted embryos in the biopsied group (49.31%) was significantly lower than that of the control (60.91%) and normal groups (96.24%) at 7.5 dpc. In addition, the survival rate of control group embryos was significant lower than that of normal group embryos. Classification of the precise developmental stages of randomly selected live implanted embryos at 7.5 dpc revealed no differences among the three groups. Our results indicate that blastomere biopsy does not adversely affect embryo implantation. The PGD procedure, in particular blastomere biopsy, increases the rate of embryo death at 4.5–7.5 dpc, but does not affect the development of surviving 7.5 dpc embryos. Anat Rec, 2012. © 2012 Wiley Periodicals, Inc.     

冰冻切片免疫过氧化物酶染色法在抗细胞性单克隆抗体筛选和鉴定中的应用

本文介绍一种快速准确地筛选和鉴定抗细胞性单克隆抗体(单抗)的冰冻切片免疫过氧化物酶染色技术。它既能筛选融合后的大量单抗(一般在3小时以内能筛选200～400孔杂交瘤细胞上清液);又可根据组织固有结构及细胞分区分布的特性,作为全面鉴定单抗的重要方法。我们运用此法从12次融合中,筛选并经克隆化获得291个单抗,进一步全面鉴定后得到抗T细胞等101个单抗。此外还讨论了消除非特异性染色的体会。     

聚D,L-乳酸/羟基磷灰石复合纤维的力学性能研究

采用熔融纺丝法制备PDLLA／HA复合纤维，探讨PDLLA／HA复合纤维的力学性能及影响因素和性能变化规律。实验结果表明：在分子量为12万的PDLLA中，加入一定量4～20μm的HA颗粒能提高复合纤维的力学性能。在PDLLA基体中添加HA的质量分数以10％为宜，以此配比制备的复合纤维的断裂强度高于其它配比复合纤维的断裂强度。采用分子量为20～30万的PDLLA制备的复合纤维断裂强度高，性能优异。复合纤维断裂强度随纤维直径的增加而下降，在直径为40～60μm时，复合纤维断裂伸长率高，弹性好。     

一金颗粒对动物小肠与急性胰腺炎的药理作用

目的：观察一金颗粒的药效学作用。方法：通过不同动物观察一金颗粒对小肠与急性胰腺炎的药理作用，采用（1）墨汁法观察一金颗粒对小鼠肠蠕动的作用；（2）离体肠肌试验观察一金颗粒对家兔离体肠肌的作用；（3）毛细血管通透性试验观察一金颗粒对小鼠毛细血管通透性的作用；（4）3％牛磺胆酸钠胰腺被膜下注射复制急性胰腺炎模型，观察一金颗粒对急性胰腺炎大鼠的治疗作用。结果：（1）小鼠肠内墨汁的推进率在大、中剂量组明显高于阴性对照组，而小剂量组略有增加；（2）能增加肠肌蠕动的幅度和频率，其中以频率增加为主；（3）各剂量组均可使小鼠毛细血管通透性降低；（4）3、6h各剂量组均可使血淀粉酶较模型有显著降低。结论：一金颗粒具有一定的增加肠蠕动．抗炎及抗胰腺炎的作用。     

固定义齿联合活动义齿修复牙本质发育不全患者1例

牙本质发育不全（DI）是一种遗传性疾病,患者牙本质发生病变,尤其多发于恒牙。患牙临床表现为棕黄色、釉质缺失、过度磨耗等,修复治疗这类牙具有极高的挑战性。本文报道1例用固定义齿联合活动义齿修复牙本质发育不全的患者。     

经腹彩色多普勒超声血流显像诊断宫内残留物的价值

目的　探讨经腹彩色多普勒超声血流显像 (CDFI)诊断宫内残留物的临床价值。方法　应用经腹超声结合 CDFI诊断宫内残留物 183例 ,并与尿 HCG和清宫术后病理结果对照。结果　 183例宫内残留物全部经清宫术后病理证实。尿 HCG阳性 171例 ,占 93.4 % (171/ 183) ,阴性 12例 ,占 6 .6 % (12 /183)。CDFI检测到残留物基底部与局部宫壁组织内有血流信号 15 8例 ,占 86 .3% (15 8/ 183) ,尿 HCG均为阳性 ;无血流信号 2 5例 ,占 13.7% (2 5 / 183) ,尿 HCG阳性 13例 ,阴性 12例。脉冲多普勒 (PW)呈类滋养层血流频谱 ,阻力指数 (RI)平均值 0 .4 4(0 .34～ 0 .5 8)。结论　经腹超声结合 CDFI能对宫内残留物进行诊断和鉴别诊断 ,并能根据血流信号检出部位进行定位 ,具有重要的临床应用价值     

乳腺浸润性导管癌超声征象与雌激素受体表达的相关性初探

目的探讨二维超声预测乳腺浸润性导管癌雌激素受体表达水平的可能性。方法回顾性分析96例经手术病理证实的乳腺浸润性导管癌的二维超声征象,并与术后标本免疫组织化学染色的雌激素受体表达水平进行对比研究。结果本组雌激素受体阳性率为55.2%(53/96),具有毛刺征(蟹足征)、周边高回声晕、后方声衰减征象之一者,肿块雌激素受体阳性率显著高于不具有上述征象的肿块(P<0.01);无微钙化组雌激素受体阳性率比微钙化组略高,但差异无显著性意义(P>0.05)。结论乳腺浸润性导管癌二维超声征象可从一定程度上反映雌激素受体表达水平。