白血病

In this topic we detected the expression level of SDF-1,CXCR4,VEGF of the above two malignant hematological tumors as well as their relations to the tumor cell"s characters of drug resistance and infiltration etc, and then elucidated the change of the microenviroment" on the occurrence and development of hematologic tumor. This topic may provide some theoretical and clinical proofs for finding out some new therapeusis to the above two malignant hematological tumors.

本课题旨在通过研究急性白血病和恶性淋巴瘤患者SDF-1、CXCR4、VEGF的表达水平及其与肿瘤细胞迁移、耐药、浸润的关系，阐明骨髓造血微环境和恶性淋巴瘤局部病变微环境的改变在上述血液肿瘤发生发展中的作用及其与血管新生的联系，为从新的角度探索白血病、淋巴瘤的治疗方法提供理论及临床依据。

Results1. Content of arsenic in serum raises with dose adding, time of pouring medicine and time of blood sampling prolonging. Thin layer chromatography indicates that there is toadfish in medicine serum of Liu-Shen-Wan.2. The suppressive rate of medicine serum of Liu-Shen-Wan on HL-60 cell raises with adding of dose and time of pouring medicine and prolonging of blood sampling.3. Gimsa dying assay, in situ end-labeling method and flow cell machine all exhibits: HI-60 cells dealed with Liu-Shen-Wan medicine-serum present typical apoptosis morphology.

结果：1 血清砷含量随着给药剂量、给药天数增加和采血时间的延长而升高；薄层色谱表明六神丸含药血清中有蟾酥成分。2 六神丸含药血清对白血病细胞的抑制率，随给药剂量、天数增加和采血时间的延长而升高3 吉姆萨染色、流式细胞仪检测、原位末端标记检测均证明六神丸含药血清能诱导HL-60白血病细胞凋亡。

Cases indicated middle cell unimodal distribution and expanded from middle to small as well as large cell areas, and 27.0% cases indicated abnormally large cell unimodal distribution） while remaining 18.9% cases showed bimodal distribution.

急性淋巴细胞白血病，白细胞直方图均表现为小细胞峰极度增高，呈单峰分布并向中间细胞区扩展变宽；急性髓系细胞白血病，81.1％白细胞直方图呈单峰分布（其中54.1％表现为中间细胞单峰增高并向小细胞区和大细胞区扩展，27.0％以大细胞单峰极度增高），18.9％呈双峰分布。

Immunohistochemical method was used to detect avian leukosis virus subgroup J from egg-type chicken, which was preliminarily diagnosed to appear subgroup J avian leukosis. The organs from these chickens were examined by ALV-J gp85 monoclonal antibody, these organs included marrow, liver, spleen, kidney, lung, heart, pancreas, oviduct, ovary, ventriculus glanduaris, skeletal muscle, cerebrum, ischiacticus nerve.

采用免疫组化法，对病理学初步诊断为蛋用型鸡 J亚群白血病的自然发病鸡的肿瘤、骨髓、肝脏、脾脏、肾脏、肺脏、心脏、胰脏、输卵管、卵巢、腺胃、骨骼肌、大脑、坐骨神经，用特异性抗 J亚群禽白血病病毒囊膜糖蛋白gp85的单克隆抗体进行检测，待检的组织切片中均检出阳性抗原，免疫组化的研究结果与病理学诊断结果相一致。

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体；2）利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达；3）构建ZNRD1的小干扰RNA载体，并测序鉴定；4）利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞（AGS、SGC7901、MKN28）和小鼠成纤维细胞（NIH3T3），G418筛选后进行鉴定；5）利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞（SGC7901）、正常胃组织上皮细胞（GES-1）、对长春新碱耐药的胃癌细胞（SGC7901/VCR）、药敏白血病细胞（HL-60）、对长春新碱耐药的白血病细胞（HL-60/VCR），G418筛选后进行鉴定；6）利用MTT实验检测ZNRD1高/低表达对细胞（AGS、SGC7901、MKN28、NIH3T3、GES-1）生长的影响；7）通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响；8）通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响；9）通过流式细胞仪分析ZNRD1高/低表达对细胞（AGS、MKN28、NIH3T3、GES-1）的细胞周期的影响；10）通过流式细胞仪分析上调ZNRD1对细胞（AGS、MKN28、NIH3T3）的凋亡的影响；11）通过MTT实验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）体外药物敏感性的影响；12）通过肾包膜下移植法检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR）体内药物敏感性的影响；13）通过流式细胞仪分析ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）内阿霉素蓄积和泵出的影响；14）通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响；15）通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60）凋亡敏感性的影响；16）通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响；17）利用RT-PCR、Western blot对基因芯片的结果进行鉴定；18）利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用；19）利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用；20）利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响；21）利用反义核酸技术抑制p21的表达；通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响；22）利用反义核酸技术抑制p27的表达；通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响；23）利用脂质体转染法上调Skp2的表达；通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响；24）利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响；25）利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响；26）利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响；27）利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞（SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR）多药耐药表型的影响；利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

Clinically, most MDS patients transform to acute myeloid leukemia, but a few MDS patients transform to aute lymphocytic leukemia, which indicates multipotential stem cells were affected in a few MDS.

临床也发现，多数MDS发展为急性髓系白血病，但也有少数演变为急性淋巴细胞白血病，提示少数MDS病变发生于多能干细胞。

"Leukemic cells that resist asparaginase and survive in this protective niche of the bone marrow might be the reason that leukemia recurs in some children who have been treated with this drug," said Dario Campana, MD, PhD, Oncology and Pathology.

Dario Campana，肿瘤学和病理学的医学博士和理学博士，说：&白血病细胞抵抗天冬酰胺酶的效果，从而在这种保护性的小生境中存活下来可能是一些接受了天冬酰胺酶治疗的儿童白血病复发的原因。&

Rece ntly new chemotherapeutic drugs always appear endlessly, and plans of chemothera py update endlessly. But the total catabatic rate and viability are still low. T he main reason is the multidrug resistance of leukemic cells to chemothera peutic drugs.

近年来新的化疗药物不断涌现，化疗方案不断更新，然而白血病总的完全缓解率和长期生存率仍然低下，究其原因，主要是白血病细胞对化疗药物的多药耐药。

Furthermore, we investigate several main cytogenetical subtypes of subacute myeloid leukemia. We aim to provide clinical and cytogenetical evidence for diagnosis and classification of subacute myeloid leukemia.

此外，还进一步探讨了亚急性髓性白血病的几类主要的细胞遗传学亚型的特征，为亚急性髓性白血病提供更科学的诊断标准和分型依据。

Cytogenetical evolution was always associated with disease progression in subacute myeloid leukemia patients.

亚急性髓性白血病在病程中出现核型演进时，均伴有病情进展，多为进展到显著的白血病阶段。

But we don't care about Battlegrounds.

但我们并不在乎沙场中的显露。

Ah! don't mention it, the butcher's shop is a horror.

啊！不用提了。提到肉，真是糟透了。