急性杂合细胞性白血病的骨髓细胞形态学读片及特点总结

目的：探讨急性杂合细胞性白血病（hybrid acute leukemia, HAL）患者骨髓细胞的形态学特点及诊断特征。方法2008年—2010年住院的急性杂合细胞性白血病患者共10例，选择其中5例典型病例及特殊病例，取其骨髓片经瑞氏染色置于OLYMPUS CX31显微镜下观察。结果杂合细胞白血病骨髓特点主要表现为细胞增生明显活跃至极度活跃，骨髓原始细胞的比例较高，细胞分化程度均较差；细胞胞体可以出现大小不等的表现，胞核多为圆形或类圆形，核染色质细致，但是细致程度粗细不等，核仁隐显不一；胞浆量多偏少，除个别特殊病例外，多不见颗粒或颗粒较少。结论双表型杂合细胞白血病患者由于骨髓细胞形态学既具有淋系特点，又具有髓系特征，因此诊断时需紧密结合骨髓细胞的免疫表型分析。

Ph^(1+)急性杂合细胞白血病附二例分析

目的 观察Ph1+ 急性杂合型白血病 (HAL)临床特征。方法 形态学检查 :骨髓涂片经瑞特染色后分类计数 ,常规进行髓过氧化酶等细胞化学染色。免疫分型 :骨髓单个核细胞悬液采用活细胞单色间接免疫荧光标记 ,流式细胞仪检测。细胞遗传学分析 :骨髓标本采用直接法及 2 4小时短期培养法处理标本 ,显带分析应用改良的热处理姬姆萨R显带 (RHG)法 ,核型分析根据ISCN(1995 )描述。治疗采用标准方案并作定期随访。结果 病例 1用标准治疗方案治疗 ,预后差。病例 2用标准治疗方案诱导缓解时间短 ,持续缓解时间长 ,预后较好。结论 根据法、英、美急性白血病诊断标准 (FAB)诊断为急性淋巴细胞白血病 ,形态学、免疫学、遗传学分型法 (MIC)诊断为Ph1+ HAL治疗效果差 ,预后恶劣。FAB诊断为急性粒细胞白血病M 2a型 ,MIC诊断为Ph1+HAL治疗效果好 。

与衰老相关的线粒体功能严重缺陷的胞质杂合细胞的分子特征

该研究组先前建立了含有不同年龄组小鼠神经细胞线粒体的胞质杂合细胞株(其细胞核基因组背景相同),发现老年组较年轻组的胞质杂合细胞线粒体总体功能显著降低。为了研究与衰老相关的线粒体功能严重缺陷的胞质杂合细胞的分子特征,该研究采用3个线粒体总体功能极其低下的老年组胞质杂合细胞株(1个正常功能年轻组胞质杂合细胞株作为对照)作为研究对象。首先,证实氧耗水平和ATP合成显著降低(P<0.05或P<0.01)。然后,对线粒体DNA(mitochondrial DNA,mt DNA)进行了高通量测序,特别是检出突变型比例低的mt DNA异质性突变,并进一步对呼吸链复合体依赖性氧耗进行了检测。测序结果显示,在老年组胞质杂合细胞中mt DNA点突变明显积累。这些突变包括非编码区的3个变异,据DNA保守性分析结果,其中2个(异质性m.15849G>T、m.16289A>T)可能为有害的;编码区的4个变异,据DNA和氨基酸保守性分析及蛋白质功能预测结果,其中2个(ND5基因的同质性m.12496T>C、Cyt b基因的异质性m.15199A>T)可能为有害的。进一步研究结果显示,同时具有复合体I亚基ND5(或复合体III亚基Cyt b)突变和2个调控区突变的胞质杂合细胞,其复合体I(或复合体III)依赖性呼吸显著降低(P<0.05或P<0.01)。综上,发生于老年组胞质杂合细胞的线粒体总体功能异常,其原因可能为,mt DNA调控区和编码区的异质性和同质性突变,以及多重mt DNA突变的累加引起线粒体呼吸链复合体功能的缺陷,进而导致线粒体总体功能异常,从而促进衰老。

在细胞含质体数目不均、分裂不同步条件下突变质体随细胞分裂而传递的模式

本文在细胞质体数目不均等、分裂不同步条件下建立了突变质体随细胞分裂而传递的数学表达式，并在具体给定条件下给出了具体计算原始细胞分裂n次后其子细胞含有x个质体，且其中已有j个质体发生突变的可能性（即概率）的公式。

在细胞周期中质体可多次分裂条件下质体随细胞分裂而传递的数学模式

本文对在细胞周期中质体分裂次数不等，细胞含有质体数目不均的复杂条件下建立了质体随细胞分裂而传递的一般性数学模式，并在每个质体都可能分裂和等可能重新随机分配条件下据此公式推出了下述结论：任何杂合细胞随着分裂次数的增加，必然出现两极分化，即纯合细胞占总体比例越来越大且趋于１００％；杂合细胞占总体的比例越来越小，且趋于０。

含两类质体的细胞随分裂而质体纯化的特征

本文先通过转移概率给出了刻划杂合状态起始细胞可纯化，单向纯化，双向纯化的特征，然后引入了临界平均后代数概念，并用此概念刻划了单向纯化、双向纯化的实质．

细胞分裂中质体的分配方式及传递规律

本文综述了质体的几种分配方式，建立了质体传递的基本数学模式，探讨了演变趋势以及杂和细胞（既含突变质体又含非突变质体的细胞）通过选择培育性状稳定的新类型的实质条件。

Regeneration of Diploid Intergeneric Somatic Hybrid Plants Between Microcitrus and Citrus via Electrofusion

Leaf-derived protoplasts of Rough lemon (Citrus jambhiri Lush, 2n = 2x = 18) were electrofused with embryogenic suspension protoplasts of its relative, Microcitrus papuana Swingle (2n = 2x = 18), with an intention of creating novel germplasm. Six plants were regenerated following protoplasts fusion. Cytological examination demonstrated that they were diploids with 18 chromosomes (2n = 2x = 18). RAPD (random amplified polymorphic DNA) analyses with six arbitrary 10-mer primers showed that the regenerated plants had identical band patterns to those of Rough lemon for primers OPA-07, OPAN-07, OPE-05 and OPA-08, Whereas for the other two primers, OPA-04 and OPS-13, bands specific to M. papuana could be detected in the regenerated plants. Cytological and RAPD analysis revealed that the regenerated plants were diploid somatic hybrids between M. papuana and Rough lemon. The putative hybrids were morphologically similar to Rough lemon. This is the first report on production of diploid somatic hybrid plants between citrus with its related genus via symmetric fusion.

Intergeneric Somatic Hybrid Plants Between Citrus and Poncirus trifoliata and Evaluation of Their Root Rot Resistance

Protoplasts of Page tangelo (Citrus reticulata Blanco×C. paradisi Macf.) cell suspension culture were electrically fused with mesophyll protoplasts isolated from trifoliate orange (Poncirus trifoliata (L.) Raf.). More than 150 plantlets regenerated after 4-5 months of culture. The regenerated plants were trifoliate with well developed root systems. Root_tip chromosome counting of more than 20 randomly selected plants revealed that they were all tetraploids (2n=4x=36). RAPD analysis of 7 randomly selected plants verified their hybridity. Inoculation of citrus Phytophthora parasitica Dastar toxin on leaves of somatic hybrids and both parental genotypes showed that Page tangelo was moderately susceptible, and trifoliate orange was highly resistant while the somatic hybrids were resistant. The potential of this somatic hybrid as rootstock is also discussed.

HLA-DQB1-PCR-SSP方法的建立及其在骨髓移植中的应用研究

为了改进骨髓移植中供、受体配型技术,在已有技术的基础上改进、建立了新的HLA-DQB1-PCR-SSP方法。利用澳大利亚皇家医院赠送的HLA标准分型细胞对自行设计引物的特异性进行鉴定,发现该引物设计合理、扩增特异。通过内部阳性对照引物的设立,可较容易地将纯合子及杂合子分型细胞分开,并对个体DNA进行了尝试。结论表明,HLA-DQB1-PCR-SSP快速、简便、灵敏、重复性好,结果易于判断,适于小样本及急症病人的器官及骨髓移植配型,是比较理想的HLA-Ⅱ类基因分型方法。

Microsatellite Alterations on Chromosome 8 of Hepatocellular Carcinoma

Objective: To study the features of microsatellite alterations on chromosome 8 and their asso- ciation with clinicopathological characteristics of hepatocellular carcinoma (HCC). Methods: Ten highly- polymorphic microsatellite markers on chromosome 8 were selected to be detected for loss of heterozygosity (LOH), microsatellite instability (MSI) and allelic imbalance (AI) in 56 HCC using MegaBACE 500 auto- matic DNA analysis system. Results: LOH was found in 37 of 56 HCC (66.1%) on at least 1 locus, and the top three loci were D8S261(53.5%), D8S1721(52.5%) and D8S1771(52.5%). LOH frequency on D8S277 was signi?cantly higher in cases positive for serum HBsAg than in those negative for HBsAg (P <0.01). Similarly, LOH on D8S261, D8S298 and D8S1733 occurred more frequently in patients with negative HB- sAg than those with positive HBsAg (P <0.01). LOH on D8S298 and D8S1771 was more frequent in those tumors larger than 3 cm in size (P <0.05 or P <0.01). LOH frequencies of D8S1721 were signi?cantly higher in the patients with absent or not intact tumor capsule than in those with intact tumor capsule (P <0.05). LOH on D8S298 and D8S1771 was more frequently detected in tumors with intrahepatic metastasis than in those without intrahepatic metastasis (P <0.01). MSI was found in 12.5% (7/56) cases, and AI was found in 19.6% (11/56), no correlation was found between MSI and AI and clinicopathological character- istics of HCC. Conclusion: Frequent microsatellite alterations on chromosome 8 existed in HCC. LOH, which represents tumor suppressor gene pathway, plays a more important role in hepatocarcinogenesis; MSI representing mismatch repair gene pathway ranks next. LOH at a particula locus is associated with the clinicopathological parameters of human HCC.

Antitumor immunopreventive and immunotherapeutic effect in mice induced by hybrid vaccine of dendritic cells and hepatocarcinoma in vivo

AIM: To develop atumor vaccine by fusion of H22 hepatocarcinoma cells and DC, and to study its protective and therapeutical effect against H22 cell.METHODS: H22-DC vaccine was produced by PEG fusion of H22 and DC induced by cytokine released from splenic mononuclear cells, sorted by CD11c magnetic microbead marker. It was injected through the tail vein of the mice and the H22-DC oncogenesis was detected in the liver, spleen and lung. In order to study the therapeutical and protective effect of H22-DC against tumor H22, two groups were divided:immune group and therapeutic group. Immune group was further divided into P, D, HD and H subgroups, immunized by PBS, DC, H22-DC and inactivated H22, respectively, and attacked by H22 cell. The tumor size, tumor weight, mice survival time and tumor latent period were recorded and statistically analyzed; Therapeutical group was divided into three subgroups of P, D and HD, and attacked by H22, then treated with PBS, DC, and H22-DC, respectively. Pathology and flow cytometry were also applied to study the mechanism how the H22-DC vaccine attacked on the H22 cell.RESULTS: 1. No oncogenesis was found in spleen, lung and liver after H22-DC injection. 2. Hybrid vaccine immunized mice had strongest CTL activity. 3. In the immune group,latent period was longer in HD subgroup than that in P, H and D subgroup; and tumor size and weight were smaller in HD subgroup than that in P, H and D subgroup. 4. In therapeutic group, tumor size was smaller in HD subgroup than that in P, D subgroup.CONCLUSION: 1. H22-DC tumor vaccine is safe without oncogenesis in vivo. 2. Hybrid vaccine can stimulate potent specific CTL activity against H22.3. H22-DC vaccine has distinctive prophylatic effect on tumor H22 and can inhibit the tumor growth.

Analysis of Heterosis, Combining Ability and Heritability of Cadmium Content in Brown Rice of Three-line Indica Hybrid Rice

Five cytoplasmic male sterile （CMS） lines were used as parents in an incomplete diallet cross and six restorer lines of rice design （Nc II design）. Thirty hybrid combinations with the same growth period were selected as experimental ma- terials to investigate the heterosis, combined ability and heredity of Cd content in brown rice of indica hybrid rice. According to the results, Cd content in brown rice showed a significantly negative heterosis; the general combining ability and specific combination ability of Cd content in CMS and restorer lines both reached extremely significant level （P〈0.01）, indicating that both genetic improvement of parents and e- valuation of combinations are important to the breeding of hybrid combinations with low accumulation of Cd; the broad-sense heritabitity and narrow-sense heritability of Cd content were both relatively high with slight differences, which respectively reached 97.73% and 80.10%, indicating that Cd content in brown rice mainly de- pends on the additive action of genes; in addition, parent improvement showed bet- ter effect on the selection of early generation.

Clinicopathological significance of loss of heterozygosity and microsatellite instability in hepatocellular carcinoma in China

AIM: To determine the features of microsatellite alterations and their association with clinicopathological characteristics of hepatocellular carcinoma (HCC). METHODS: Loss of heterozygosity (LOH) and microsatellite instability (MSI) of 55 microsatellite loci were detected with PCR-based microsatellite polymorphism analyses in tumors and corresponding noncancerous liver tissues of 56 surgically resected HCCs using the MegaBACE 500 automatic DNA analysis system.RESULTS: LOH was found in 44 of 56 HCCs (78.6%) at one or several loci. Frequencies of LOH on 1p, 4q, 8p,16q, and 17p were 69.6% (39/56), 71.4% (40/56), 66.1% (37/56), 66.1% (37/56), and 64.3% (36/56), respectively. MSI was found in 18 of 56 HCCs (32.1%) at one or several loci. Ten of fifty-six (17.9%) HCCs had MSI-H. Serum HBV infection, alpha-fetoprotein concentration, tumor size, cirrhosis, histological grade, tumor capsule, as well as tumor intrahepatic metastasis, might be correlated with LOH on certain chromosome regions. CONCLUSION: Frequent microsatellite alterations exist in HCC. LOH, which represents a tumor suppressor gene pathway, plays a more important role in hepatocarcinogenesis. MSI, which represents a mismatch repair genepathway, is a rare event during liver carcinogenesis. Furthermore, LOH on certain chromosome regions may be correlated with clinicopathological characteristics in HCC.

Correlation between expression and differentiation of endocan in colorectal cancer

AIM: To investigate the expression frequency of endocan in colorectal cancer and analyze the relationship between endocan expression and clinical parameters and to study the role of endocan in colorectal carcinogenesis. METHODS: Expression of endocan in 72 tumor tissue samples of colorectal cancer as well as in 27 normal mucous membrane tissue samples was analyzed using in situ hybridization, immunohistochemistry on tissue microarray, Western blot and reverse-transcript polymerase chain reaction (RT-PCR). RESULTS: The expression of endocan was higher in normal colon and rectum tissue samples than in cancerous tissue samples (mRNA = 92.6%, protein = 36%), and was lower in colorectal cancer tissue samples (mRNA = 70.4%, protein = 36.1%). No correlation was found between staining intensity and clinical parameters such as sex, age, tumor size andTNM stage. However, the expression of endocan was positively correlated with the tissue differentiation in colorectal cancer. CONCLUSION: The expression of endocan is down- regulated in colorectal cancer and is positively correlated with the tissue differentiation in colorectal cancer, suggesting that the expression of endocan is associated with development and differentiation of colorectal cancer.

Cloning and characterization of a novel gene (C17orf25) from the deletion region on chromosome 17p13.3 in hepatocelular carcinoma

Using a combination of hybridization of PAC to a cDNA library and RACE technique, we isolated a novel cDNA, designated as C17orf25 (Chromosome 17 open rea(ling frame 25, previously named it HC71A), from the deletion region on chromosome 17p13.3. The cDNA encodes a protein of 313 amino acids with a calculated molecular mass of 34.8 kDa. C17orf25 is divided into 10 exons and 9 introns, spanning 23 kb of genomic DNA. Northern blot analysis showed that the mRNA expression of C17orf25 was decreased in hepatocellular carcinoma samples as compared to adjacent noncancerous liver tissues from the same patients. The transfection of C17or25 into the hepatocellular carcinoma cell SMMC7721 and overexpression could inhibit the cell growth. The above results indicate that C17orf25 is a novel human gene, and the cloning and preliminary characterization of C17orf25 is a prerequisite for further functional analysis of this novel gene in human hepatocellular carcinoma.

THE RELATIONSHIPS BETWEEN ERYTHROBLAST DENUCLEATION AND THE NUCLEAR MATRIX－INTERMEDIATE FILAMENTS

We present a novel approach for making cybrids. By introducing neo gene expression plasmids into rabbit reticulocytes, fusing the gene transferred reticulocytes with K562 cells and selecting in G418 selection medium, a cybrid strain K-RRneo was established. Whole mount TEM study demonstrated that after cybridization, there was a reorganization of the intermediate filaments which showed a tendency to differentiate towards reticulocytes. SDS-PAGE and western blot analysis verified the above observation, in which the vimentin blot pattern of the cybrids was similar to that of reticulocytes, but totally different from that of K562 cells. Using this model, we reaffirmed the hypothesis that the erythroid differentiation factor (EDF) might be responsible for erythroid differentiation as well as the initiation of denucleation.

Progress and future development of hybrid rapeseed in China

The hybrid growth of rapeseed now has reached to 4.7×106hm2,accounting for 70 % of total rapeseed growth area in China. Over the last 40 years,the researches and utilizations of hybrid rapeseed in China have significantly contributed to the development of rapeseed industry in China. The production of hybrid rapeseed mainly utilizes cytoplasmic male sterility(CMS),combined at the same time with nuclear male sterility and other pollinating-control systems in China. The hybrid rapeseed studies in China are also characterized by the improvement of quality and oil content in hybrid breeding. Future studies to enhance the heterosis of rapeseed will be focused on several important issues,including the combination of heterosis and ideotype breeding,further increase of oil content in hybrids,utilization of sub-genomic heterosis and resistance improvement. The paper will discuss the following perspectives in hybrid rapeseed studies:relationships among heterosis,quality and disease traits,solutions for excessive source and pool in hybrids compared with open-pollinated cultivars,and the importance of increasing harvest index of hybrids to achieve a better yield in hybrids.