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.     

Apatinib in gastric carcinoma: A case report of partial response for first‐line treatment in advanced disease

Gastric cancer (GC) is the most common gastrointestinal malignant tumor, with a gradual increasing incidence throughout the world. Mostly GC is diagnosed in its late stage. To date, there is no usable standardized treatment regimen for patients with advanced GC. Apatinib mesylate, small‐molecule vascular endothelial growth factor receptor‐2 (VEGFR‐2) tyrosine kinase inhibitor (TKI), has been approved as third‐line treatment for patients with advanced gastric adenocarcinoma in China, October, 2014. Till now, there is no case report about apatinib as first‐line treatment for patients with advanced GC in literature. We present an 83‐year‐old Chinese man with advanced gastric adenocarcinoma, who received apatinib as first‐line option and obtained clinical benefit within 2 weeks. The lung metastases disappeared completely and the liver metastases shrank significantly. The patient's progression‐free survival was 163 days and overall survival was 201 days. This paper reviews and discusses apatinib as a new targeted drug for patients with advanced GC by comparison with other effective molecular‐targeted therapy.     

Initial Progression-Free Survival after Non-First Line TKIs Therapy Potentially Guides Immediate Treatment after Its Failure in Advanced Non-Small Cell Lung Cancer

Objective The standard therapy after failure of the initial non-first line epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) treatment in advanced non-small cell lung cancer(NSCLC) has not yet been established.The aim of the current study was to identify whether the 2 TKI treatment or chemotherapy(paclitaxel-containing or non-paclitaxel regimen) is the appropriate treatment for patients with NSCLC based on the efficacy of the initial TKIs. Methods Seventy-two advanced NSCLC patients who had accepted 2 TKIs or chemotherapy immediately after failure of the initial TKIs in non-first line setting from May 1,2004 to January 31,2010 at the Sun Yat-sen University Cancer Center were enrolled.The primary endpoint[2 progression-free survival(PFS)]and the second endpoint[overall survival(OS)]were compared among the 2 TKI and chemotherapy groups as well as their subgroups. Results(1) Twenty-one patients were treated with 2 TKIs,and 51 patients were administered chemotherapy after failure of the initial non-first line TKI treatment.There was nonsignificant difference in the responses(P=0.900)[2 PFS(P=0.833) and OS(P=0.369)] between the 2 TKI and chemotherapy groups.(2) In the 2 TKI group,9 patients exhibited PFS>7 months.The initial TKI treatment group exhibited a longer 2 PFS than the other 12 patients with an initial PFS<7 months(7 months vs.2 months,P=0.019).However, these groups had nonsignificantly different OS(P=0.369).(3) In the chemotherapy group,patients with PFS<5 months exhibited longer 2 PFS than those with PFS > 5 months in the initial TKI treatment(3 months vs.2 months,P=0.039).(4) In the chemotherapy group, patients treated with paclitaxel-containing regimen showed longer 2 PFS than those treated with non-paclitaxel regimen(5 months vs.2.3 months,P=0.043). Conclusions Patients with PFS>7 months or <5 months under the initial TKI treatment potentially benefit from the 2 TKI treatment or chemotherapy immediately after failure of the non-first line TKIs.The paclitaxel-containing regimen may improve the 2 PFS. However,more patient samples are urgently needed to validate these findings.     

雷替曲塞联合放疗治疗老年食管癌的Ⅱ期前瞻性临床研究

目的 探讨雷替曲塞联合放疗治疗老年食管癌的临床疗效及不良反应.方法 采用信封法随机将60例老年食管癌患者分成雷替曲塞同步放化疗组(试验组30例)与单纯放疗组(对照组30例),采用常规分割方式,放疗剂量56~60 Gy/28~30 F,试验组放疗第1、22天给予雷替曲塞2.6 mg/m2单药增敏化疗,放疗期间连用2周期,观察两组患者的近期疗效、生存时间及不良反应发生情况.结果 试验组总有效率为93.3%,而对照组为73.3%,差异有统计学意义(χ2=4.320,P=0.038).试验组及对照组中位生存时间分别为24.0个月、12.0个月,经Log-rank检验差异有统计学意义(χ2=6.048,P=0.014).试验组和对照组患者常见的重度不良反应(3~4级)如放射性食管炎(10.0%∶3.3%;χ2=0.268,P=0.605)、白细胞减少(13.3%∶10.0%;χ2=0.000,P=1.000)、血小板减少(3.3%∶0;P=1.000)、恶心呕吐(6.7%∶0;χ2=0.517,P=0.472)发生率的差异均无统计学意义.结论 雷替曲塞联合放疗治疗老年食管癌可显著提高其近期疗效,延长生存期,且不良反应轻微,值得进一步临床研究.     

以地西他滨为主方案治疗初发老年人急性髓系白血病近期疗效分析

目的 探讨以地西他滨为主方案治疗初发老年人急性髓系白血病(AML)的临床安全性和有效性.方法 选择2014年6月至2015年12月首都医科大学附属北京潞河医院收治的12例初发老年AML患者,应用地西他滨单药或联合低剂量化疗方案治疗,对12例患者临床资料进行回顾性分析.结果 12例患者中1个疗程达完全缓解(CR)者6例,部分缓解(PR)者5例,未缓解(NR)者1例.6例CR患者中1例在第6个疗程后复发,其余在随访中仍CR;5例PR患者中2例经2个疗程化疗后达CR,1例在第2个疗程化疗后复发,2例在第3个疗程化疗后复发.1例患者经2个疗程化疗后NR,因肺部感染死亡.4例复杂核型患者中3例疗效差,1例达CR,但最终仍复发;其余8例+8、-X或正常核型患者中7例短期达CR.主要不良反应为骨髓抑制和感染,所有患者均能耐受.结论 地西他滨单药或联合低剂量化疗方案治疗初发老年人AML近期疗效好,耐受性好,安全性高,可作为一线治疗方案.     

肝癌Huh7干细胞样细胞的富集培养及鉴定

目的：建立一种体外有效富集、培养和鉴定具有肝癌干细胞特征的细胞亚群的方法。方法：采用成球培养法利用肿瘤干细胞样细胞（cancer stem cell, CSC）分化培养基对肝癌Huh7细胞进行富集培养,获得的干细胞样细胞于体外进一步扩增获得肝癌干细胞球。流式细胞术检测Huh7干细胞样细胞表面肿瘤干细胞标志物EpCAM、CD90和CD133的表达,平板克隆集落形成实验和裸鼠成瘤实验分别检测Huh7细胞和Huh7干细胞样细胞的克隆集落形成能力、体内成瘤能力。结果：Huh7细胞成球培养3~7 d后即可形成肝癌干细胞样细胞球,获得的干细胞样细胞具有自我更新和增殖能力,其EpCAM阳性细胞比例较Huh7细胞明显增加\[（99.6%±0.31）% vs（0.12%±0.05）%,P<0.01\],但两种细胞CD90\[(0.11%±0.06)% vs (0.09%±0.07)%, P>0.05\]和CD133\[(0.17%±0.08)% vs (0.15%±0.05)%, P>0.05]表达差异无统计学意义。Huh7干细胞样细胞克隆集落形成数量明显多于Huh7细胞\[（188.67±12.5）vs （79±16.7）个,P<0.01\];当接种量为5×104个细胞时,与接种Huh7细胞的对照裸鼠相比,接种Huh7干细胞样细胞的裸鼠成瘤时间更短（11 vs 30 d）,成瘤率更高（100% vs 16.67%）;接种5×105数量级的细胞时,实验组成瘤体积\[（171.90±10.94）vs（86.39±11.21）mm3, P<0.01\]和瘤体质量\[（2.98±0.82）vs（0.32±0.17）g, P<0.01\]均明显大于对照组。结论：利用成球培养法能够从Huh7肝癌细胞系中富集培养获得Huh7肝癌干细胞,其具有比Huh7细胞更强的成瘤能力。     

Inhibition of sorcin reverses multidrug resistance of K562/A02 cells and MCF-7/A02 cells via regulating apoptosis-related proteins

Purpose

Sorcin, a 22-kDa calcium-binding protein, renders cancer cells resistant to chemotherapeutic agents, thus playing an important role in multidrug resistance (MDR). But the mechanisms mediated by sorcin still remain quite elusive. This study aim to explore whether sorcin silencing could restore chemosensitivity in MDR cancer cells and seek to identify the functional mechanisms mediated by sorcin.

Methods

To investigate the mechanisms of sorcin-silencing-induced chemosensitivity, transient expression of sorcin-siRNAs was performed in doxorubicin-induced MDR cell lines, K562/A02 and MCF-7/A02. Sensitivity to five chemotherapeutic agents was evaluated by analysis of cell survival and cell apoptosis.

Results

In this report, we show that down-regulation of sorcin did not alter expression or function of P-gp, but actually induced cell apoptosis and chemosensitivity in K562/A02 and MCF-7/A02. We also observe that silencing of sorcin-enhanced chemotherapeutic agent effects partly through regulating apoptosis-related protein, including Bcl-2, Bax, c-jun and c-fos.

Conclusion

This offers the rationale for the development of therapeutic strategies down-regulating sorcin expression for the treatment of cancer, especially for the reversal of MDR.     

tBid基因的表达及其对骨肉瘤SOSP-9607细胞的促凋亡作用

目的：观察tBid基因在骨肉瘤细胞中的表达及其对转染的SOSP-9607细胞的凋亡作用。方法：采用脂质体转染的方法,将tBid基因转染骨肉瘤细胞,间接免疫荧光法检测目的蛋白的表达和细胞形态学变化,进一步电镜观察其超微结构改变。MTT法检测目的基因转染后细胞的增殖情况,通过Annexin V、染色流式细胞仪检测来观察其促凋亡作用。结果：转染SOSP-9607细胞后,间接免疫荧光染色检测tBid的表达,细胞出现凋亡。电镜观察呈现出典型的凋亡特征。MTT实验发现细胞的增殖被明显抑制。Annexin V染色流式细胞仪检测可见明显的凋亡细胞,与对照组细胞（凋亡率为6%）相比,其凋亡率为35.8%。结论：tBid基因可以在转染的SOSP-9607细胞中表达,并可抑制转染细胞的生长,诱导细胞发生凋亡。     

Association between RAD51 gene polymorphism (-135G/C) and susceptibility of myelodysplastic syndrome and acute leukemia: evidence based on a meta-analysis

Study results on the association between RAD51 gene -135G/C polymorphism and risk of myelodysplastic syndrome (MDS) or acute leukemia are inconsistent. A meta-analysis was conducted to identify the association. A systematic search was performed in PubMed, Embase, CNKI, VIP, Wanfang databases to collect all relevant studies until January 2013. Meta-analysis was carried out using fixed/random model by Review Manager 5.1 and STATA10.0. A total of 10 eligible studies with 2,656 patients and 3,725 controls were included in meta-analysis. Significant association was detected between -135G/C polymorphism and increased MDS risk (CC?+?GC vs. GG: OR?=?1.46, 95 % CI?=?1.11–1.92; CC vs. GC?+?GG: OR?=?2.45, 95 % CI?=?1.23–4.89), while no association was observed for acute leukemia. Subgroup analysis by subtypes of acute leukemia and ethnicity showed no significant results either. Our meta-analysis indicated that the -135G/C polymorphism might be associated with increased susceptibility of MDS. However, lack of evidence supported association of this polymorphism with acute leukemia. Additional well-designed studies with larger samples are required to verify our results.     

In vitro andin vivo chemotactic effect of mip-lα gene transfected tumor vaccine on immune effector cells

Vaccination with chemokine gene-transfected tumor cells may be a new apporach to cancer treatment. Macrohage inflammatory protein-la (MIP-lα ) is a new type of chemokines which has chemotactic activity on effector cells. In the present study, the B16 melanoma cells were transfected with recombinant adenovirus harboring murine MIP-1α gene. The biological characteristics of the MIP-lα gene transfected B16 melanoma cells were investigated. The level of MlP-lα in the supernatant of gene-transfected melanoma cells was 368±24 ng/ml/10⁶/24hr. This supernatant showed strong chematactic activity for NK cells, CD4⁺ T cells, CD8⁺ T cells or the freshly isolated peritoneal macrophages. Though thein vitro growth were not altered, the tumorigenicity of the gene-transfected B16 melanoma cells decreased signicantly. The infiltration of inflammatory cells into the tumor mass formed by MIP-lα gene-transfected B16 cells were much more obvious than that by wild-type B16 cells or B16 cells transfected with the control gene. However, the survival time of the mice bearing B16 melanoma cells transfected with MIP-1 a gene was not prolonged and the NK, CTL activity remianed unchanged. We suggested that thein vivo phenomenon may be related to the high toxicity of the MIP-1 α as a strong non-specific inflam-matory mediator.