Immortalized mouse fetal liver stromal cells support growth and maintenance of human embryonic stem cells

Human embryonic stem cells (hESCs) are usually maintained in an undifferentiated state by co-culture with feeder cells. The feeder cells are important for the growth of hESCs. A novel spontaneously immortalizated mouse fetal liver stromal cell line, named KM3, was isolated from a?13.5?day mouse fetal liver. In this study, we examined whether KM3 cells could be used as feeders to support the growth of hESCs. hESCs cultured on KM3 cells showed a similar proliferation rate and characteristics to mouse embryonic fibroblasts?(MEFs) after prolonged culture, including morphology, unlimited and undifferentiated proliferative ability, maintenance of normal karyotypes, formation of embryoid bodies in?vitro and typically immature teratomas in?vivo. Our results indicate that the immortalized KM3 cell line has the potential to support the growth and maintenance of hESCs. The cell line may be used for the large-scale expansion of hESCs in a low-cost and less labor-intensive manner.     

New alternatively spliced variant of prostate-specific membrane antigen PSM-E suppresses the proliferation, migration and invasiveness of prostate cancer cells

PSM-E is a newly discovered alternatively spliced variant of prostate-specific membrane antigen (PSMA). In the current study, its role on the proliferation, invasiveness and migration in prostate cancer cell lines was analyzed. PSM-E and PSMA (as a comparison) eukaryotic expression vectors pcDNA3.0/PSM-E and pcDNA3.0/PSMA were constructed, validated by RT-PCR and Western blotting, and PSMA/PSM-E overexpression PC-3 cell models were built. Gene interference was used to block PSMA and the expression of its splice variants in LNCap cells. Three shRNA fragments were synthesized against PSMA, cloned into the vector pSilencer?2.1-U6-neo, their interference effect was evaluated by RT-PCR and Western blotting, and pSilencer?2.1-U6-neo?shRNA3 (named p?shRNA3) was chosen in further analyses. Growth curves were drawn to observe the proliferation change, which showed that PSM-E had the potential to suppress proliferation (P<0.05), but no significant change was observed in PSMA/PC-3 cells and in PSMA/PSM-E interfering LNCap cells (P>0.05). Cross-river test showed that the migration speeds of PSM-E/PC-3 and PSMA/PC-3 were both significantly slower than the vector negative control, and faster in p-shRNA3 interfering LNCap cells compared with its vector negative control (P<0.05), and no significant difference existed between PSM-E/PC-3 and PSMA/PC-3 (P>0.05). Transwell assay showed that the invasive cells of both PSMA/PC-3 and PSM-E/PC-3 were fewer compared to the vector negative control (P<0.05), and the invasive suppression effect of PSM-E was weaker than PSMA (P<0.05), and accordingly, invasiveness of interfering LNCaP cells was enhanced compared with the vector negative control (P<0.05). These results showed that PSM-E could suppress proliferation, migration and invasiveness of prostate cancer cells. Its suppression effect on cell proliferation is stronger compared to PSMA and the suppression effect on invasiveness is weaker than that of PSMA.     

Racial/ethnic differences in breast cancer survival by inflammatory status and hormonal receptor status: an analysis of the Surveillance,Epidemiology, and End Results data

Background

Compared to non-inflammatory breast cancer (non-IBC), inflammatory breast cancer (IBC) has less favorable survival and is more likely to be estrogen receptor (ER) and progesterone receptor (PR) negative. ER?/PR? tumors, regardless of histology, have less favorable survival. While black women are more likely to have IBC and ER?/PR? tumors than white women, it is unclear whether the racial disparity in survival is explained by these factors. The objective of this study was to assess racial/ethnic differences in breast cancer survival by inflammatory status and hormone receptor status.

Methods

This study examined breast cancer mortality among non-Hispanic white (NHW), Hispanic white, black, and Asian/Pacific Islander (API) women diagnosed between 1990 and 2004 using the National Cancer Institute’s Surveillance, Epidemiology, and End Results data. Kaplan–Meier survival curves and Cox proportional hazard ratios (HRs) assessed the relationship between race/ethnicity and survival.

Results

Black women had significantly poorer survival than NHW women regardless of inflammatory status and hormone receptor status. Compared to NHWs, the HRs for black women were 1.32 (95 % confidence interval (CI) 1.21–1.44), 1.43 (95 % CI 1.20–1.69), and 1.30 (95 % CI 1.16–1.47) for IBC, IBC with ER+/PR+, and with ER?/PR?, respectively. Similar HRs were found for non-IBC, non-IBC with ER+/PR?, and non-IBC with ER?/PR?. API women had significantly better survival than NHW women regardless of inflammatory status and hormone receptor status.

Conclusion

Compared to NHW women, black women had poorer survival regardless of inflammatory status and hormone receptor status and API women had better survival. These results suggest that factors other than inflammatory status and hormone receptor status may play a role in racial/ethnic disparities in breast cancer survival.     

羟基喜树碱(拓僖)联合高聚生治疗恶性难治性胸腔积液

目的:观察应用羟基喜树碱联合高聚生治疗恶性、难治性胸腔积液的疗效.方法:对临床治疗中效果较差的中、大量恶性胸腔积液及既往治疗无效的恶性胸腔积液患者66例(治疗组36例,对照组30例)进行治疗,全部患者均经PICC管(外周穿刺中心静脉导管)行胸腔闭式引流,1～3天引流至胸水近乎消失后注入药物.治疗组羟基喜树碱(拓僖)30mg,高聚生2000～3 000U胸腔注射,地塞米松5mg肌肉注射;对照组顺铂60～70mg,地塞米松10mg胸腔注射,胃复安20mg肌注.结果:治疗组CR 44.4%(16/36),有效率为86.1%(31/36).对照组CR 26.7%(8/30),有效率为63.3%(19/30).结论:羟基喜树碱联合高聚生治疗恶性难活性胸腔积液疗效满意,不良反应轻微,尤其对大量血性胸腔积液及既往治疗无效者效果良好,安全、可靠,值得临床广泛应用.     

阿尔茨海默病差异表达基因的生物信息学分析

目的：应用生物信息学方法分析阿尔茨海默病(Alzheimer,s disease,AD)不同病情程度的差异表达基因（differentially expressed genes,DEGs）,在分子水平上探讨AD的发病制,为研究AD提供新思路。方法：从基因表达综合数据库(gene expression omnibus,GEO)下载基因芯片数据集GSE28146,使用在线分析工具GEO2R分别筛选AD轻度组、中度组和重度组与正常对照组比较的DEGs。运用DAVID 数据库对筛选出的DEGs进行基因本体（gene ontology,GO）富集分析和京都基因与基因百科全书（Kyoto encyclopedia of genes and genomes,KEGG）通路分析,通过STRING数据库构建蛋白相互作用网络,并采用Cytoscape软件筛选关键基因。结果：AD轻度组、中度组和重度组分别筛选出881、896和1142个DEGs。GO功能富集显示：轻度组与凋亡相关过程的激活、调节免疫反应、蛋白质磷酸化等生物过程密切相关,中度组与炎症反应、凋亡调节、钙离子的释放等生物过程密切相关,重度组与NF-κB的激活、蛋白质磷酸化和去磷酸化、细胞外基质的降解等生物过程密切相关。KEGG分析显示：轻度组主要富集到p53和TGF-β等信号通路,中度组主要富集到癌症途径、自然杀伤细胞介导的细胞毒性、细胞黏附分子等信号通路,重度组主要富集到细胞周期、Hippo信号通路、神经活性配体-受体相互作用等信号通路。蛋白互作网路分析各组富集程度最高的前5个关键基因：轻度组为GART、EHMT2、KRAS、ESR1、CD44;中度组为CBLB、HERC1、UBE2G1、UBE2M、HECW2;重度组为UBE2C、SOCS3、FBXW7、UBE3B、UBA6。结论：采用生物信息学方法分析不同病情程度的AD,所富集的信号通路和筛选出的关键基因可能在AD发生发展过程中起重要作用,为进一步深入研究AD提供了依据和思路。     

Predictive value of MMP-7 expression for response to chemotherapy and survival in patients with non-small cell lung cancer

Fundamental studies have suggested that matrix metalloproteinases-7 (MMP-7) expression is associated with chemoresistance and constitutes a prognostic factor in several solid tumors. The present study assessed the prognostic and predictive value of MMP-7 in tumors of patients with advanced non-small cell lung cancer (NSCLC) treated with platinum-based chemotherapy. In total, 159 patients with stage III and IV NSCLC were retrospectively enrolled. Immunohistochemistry was performed to evaluate the expression of MMP-7, apoptosis-related proteins Bcl-2, Bax, Fas and FasL and the Ki-67 proliferation marker. The TUNEL (terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling) method was performed to investigate tumor apoptosis. Ninety carcinomas (56.6%) were identified as high expression of MMP-7. Overexpression of MMP-7 was more frequent in adenocarcinomas than in squamous cell carcinomas (P = 0.032). The expression of MMP-7 was positively related with Ki-67 index and Bcl-2, but not apoptosis index. MMP-7 status was correlated inversely with response to chemotherapy in overall patients (response rates, 20.0% and 35.8%, for patients with high-MMP-7 and low-MMP-7 tumors, respectively, P = 0.036), especially in adenocarcinoma (P = 0.021), but not in patients with squamous cell carcinomas (P = 0.373). The overall survival was significantly lower in NSCLC patients with high MMP-7 than in those with low MMP-7 (P < 0.001). A Cox regression analyses also demonstrated MMP-7 status to be a significant prognostic factor (hazard ratio, 5.49 P = 0.001). These findings suggest that the expression level of MMP-7 in tumor cells is predictive of response to chemotherapy and outcome in patients with advanced NSCLC receiving platinum-based chemotherapy.     

食管、贲门癌围手术期细胞免疫功能动态变化的临床研究

目的：探讨食管、贲门癌全身、局部细胞免疫功能状态及生物治疗的可行性。方法：采用免疫组化观察分析５０例食管、贲门癌局部肿瘤浸润淋巴细胞（ＴＩＬ）、肿瘤相关性巨噬细胞（ＴＡＭ）的分布特征,采用ＥＬＩＳＡ法和硝酸还原酶法分别地体内可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）表达和一氧化氮（ＮＯ）生成水平进行动态检测。结果：１）癌周物浸润密度大于癌果间质;２）２术前血清ｓＩＬ－２Ｒ水平高于正常对照组,根治术后ｓ     

Acetylcholinesterase expression mediated by c-Jun-NH2-terminal kinase pathway during anticancer drug-induced apoptosis

It has been shown that acetylcholinesterase (AChE) expression was induced during apoptosis and the anti-sense oligonucleotides and siRNA of AChE may prevent apoptosis in various cell types. However, the mechanisms underlying AChE upregulation remain elusive. We demonstrated here that c-Jun NH2-terminal kinase (JNK) could mediate AChE expression. In this study, both etoposide and excisanin A, two anticancer agents, induced apoptosis in colon cancer cell line SW620 as determined by Annexin V staining, the cleavage of caspase-3 and the proteolytic degradation of poly (ADP-ribose) polymerase (PARP). The results showed that both the agents upregulated AChE in SW620 cells. In the meantime, JNK was also activated and the expression and phosphorylation of c-Jun increased in SW620 cells exposed to the two agents. The induced AChE mRNA and protein expression could be blocked by SP600125, a specific inhibitor of SAPK/JNK, and small interfering RNA directed against JNK1/2. Transfection with adenovirus-mediated dominant negative c-Jun also blocked the upregulation of AChE expression. Together, these results suggest that AChE expression may be mediated by the activation of JNK pathway during apoptosis through a c-Jun-dependent mechanism.     

射波刀治疗35例肝脏肿瘤的临床分析

目的：观察射波刀（cyberknife）治疗肝脏恶性肿瘤的疗效和安全性。方法：回顾性分析35例经射波刀治疗的肝脏恶性肿瘤患者,其中原发性肝癌12例,消化系统肿瘤肝转移16例,其他肿瘤肝转移7例。共57个病灶。所有患者行金标植入术,1周后行射波刀照射。平均肿瘤体积98.64ml,处方剂量18-51Gy,分割3-7次,等剂量线56%-85%。治疗后1-6个月复查,观察近期疗效,之后每3个月随访1次。结果：7例患者达到完全缓解,20例部分缓解,6例稳定,2例进展,有效率（CR＋PR）77.1%,局部控制率94.29%,中位TTP为12个月,中位生存期为23个月。所有患者耐受性良好,主要的不良反应为白细胞降低、食欲减退和疲劳。无Ⅳ级和Ⅳ级以上不良反应发生。结论：射波刀治疗肝脏恶性肿瘤有较高的有效率,安全性好,不良反应轻,患者易耐受。