辛伐他汀对体外培养鼠骨形成蛋白BMP-2基因的调控

目的 研究辛伐他汀对鼠骨形成蛋白(bone morphogenetic protein-2,BMP-2)及其mRNA表达的影响,验证辛伐他汀是否能够促进鼠成骨细胞的骨形成.方法 将兔干分为假手术组,模型组和辛伐他汀组3个小组,每组6只.在辛伐他汀组用药1月、2月和4月时,收集各组兔子的耳中央动脉血清,将上述血清加入鼠MC3T3-E1细胞培养4d后提取总RNA,逆转录出骨形成蛋白BMP-2的cDNA后进行实时荧光定量PCR,对产物进行检测.结果 用药1个月,辛伐他汀组BMP-2表达量较假手术组高2.79倍(P=0.0004),较模型组高2.66(P=0.001);用药2个月,辛伐他汀组BMP-2表达量较假手术组高1.88倍,较模型组高1.75倍(P=0.001);用药4个月,辛伐他汀组BMP-2表达量较假手术组高0.86倍(P=1.000),较模型组高1.04倍.结论 辛伐他汀对骨形成有正效应.     

Molecular basis of the RHD gene in blood donors with DEL phenotypes in Shanghai

Background and Objectives  The purpose of the work was to analyse the genotype of D-elute (DEL) samples and to elucidate whether there were novel DEL alleles in Chinese population.
Materials and Methods  D-negative samples were identified by an indirect antiglobulin test (IAT), and absorption∖elution tests to screen weak D, partial D and DEL phenotypes. DELs were further analysed by multiplex PCR, PCR-sequence-specific primers (PCR-SSP) and sequencing. Some of the DEL samples were determined to show RHD zygosity by PCR-restriction fragment length polymorphism or real-time quantitative PCR.
Results  Of 400 253 samples from individual donations, 1585 (0·40%) were typed as D negative. Among these D-negative samples, 279 DELs were observed. Two hundred and sixty-eight DELs were confirmed to have the RHD (K409 K) allele. Three DELs seemed to have RHD-CE-D hybrid alleles, including one RHD-CE(4–9)-D , one RHD-CE(2–5)-D and one suspected RHD(1–9)-CE . Five novel RHD alleles were found among the rest of the DEL samples, including four RHD 3 g > a, one RHD (R10W), one RHD (L18P), one RHD (L84P) and one RHD (A137E). Eighty-four DELs were analysed for Rhesus box zygosity, there were 77 RHD +/ RHD– and seven RHD +/ RHD +.
Conclusion  About 4·35% apparent D negative Chinese individuals were weak D or partial D, while 17·60% were DEL. Novel DEL alleles are rare, and all but 11 of the 279 DELs were due to the most common DEL allele, RHD (K409 K). The RHD 3G > A might be the second most frequent DEL allele in Chinese. Exploration of a complete molecular basis for DEL in Chinese ethnic groups is a long-term endeavour.     

Adaptive changes in serotonin neurons of the raphe nuclei in 5-HT(4) receptor knock-out mouse

Decreased serotonin (5-HT) transmission is thought to underlie several mental diseases, including depression and feeding disorders. However, whether deficits in genes encoding G protein-coupled receptors may down-regulate the activity of 5-HT neurons is unknown currently. Based on recent evidence that stress-induced anorexia may involve 5-HT(4)receptors (5-HT(4)R), we measured various aspects of 5-HT function in 5-HT(4)R knock-out (KO) mice. When compared to dorsal raphe nucleus (DRN) 5-HT neurons from wild-type mice, those from 5-HT(4)R KO mice exhibited reduced spontaneous electrical activity. This reduced activity was associated with diminished tissue levels of 5-HT and its main metabolite, 5-hydroxyindole acetic acid (5-HIAA). Cumulative, systemic doses of the 5-HT uptake blocker citalopram, that reduced 5-HT cell firing by 30% in wild-type animals, completely inhibited 5-HT neuron firing in the KO mice. This effect was reversed by administration of the 5-HT(1A) receptor (5-HT(1A)R) antagonist, WAY100635, in mice of both genotypes. Other changes in DRN of the KO mice included increases in the levels of 5-HT plasma membrane transporter sites and mRNA, as well as a decrease in the density of 5-HT(1A)R sites without any change in 5-HT(1A) mRNA content. With the exception of increased 5-HT turnover index in the hypothalamus and nucleus accumbens and a decreased density of 5-HT(1A)R sites in the dorsal hippocampus (CA1) and septum, no major changes were detected in 5-HT territories of projection, suggesting region-specific adaptive changes. The mechanisms whereby 5-HT(4)R mediate a tonic positive influence on the firing activity of DRN 5-HT neurons and 5-HT content remain to be determined.     

One single dose of rituximab added to a standard regimen of CHOP in primary treatment of follicular lymphoma appears to result in a high clearance rate from circulating bcl-2/IgH positive cells: Is the end of molecular monitoring near?

Accurate monitoring of the t(14;18) translocation is regarded as highly desirable in patients with follicular lymphoma (FL) as absence of bcl-2/IgH positive cells has been correlated with a reduced risk of recurrence and, more recently, pre-treatment t(14;18) load with response to rituximab (R; Blood 2004;103:4416-23). With the arrival of R clinical and molecular remission rates for various lymphoma entities improved significantly, creating the need to carefully review and reassess the role of PCR negativity for clinical outcome, specifically when considering the prolonged presence of the drug as compared to chemotherapy. To determine the rate of molecular clearance achieved by the addition of different doses of R 16 newly diagnosed, t(14;18) positive patients with FL (Ann Arbor stages III/IV) were investigated before, during and after primary chemotherapy with six cycles of CHOP combined with varying (1, 3 or 6) cycle numbers of R (varR1-, varR3- or varR6-CHOP, respectively) regarding molecular remission status. For this purpose classic nested PCR as well as a newly established RQ-PCR employing juxtaposed hybridisation probes were employed to assess molecular remission prior, during and post therapy. Interestingly, administering just a single cycle of R (varR1-CHOP) in combination with a standard regimen of CHOP resulted in rapid and effective clearance of t(14;18) carrying cells from the peripheral blood of 4/5 patients in this treatment group. 6/8 (6/8) varR1-/varR3-CHOP patients were fully cleared and 8/8 (7/8) var6-CHOP patients as assessed by RQ- (nested) PCR. This indicates the high clearance capacity of this combination therapy approach even when adding a very low cycle number of R (1 and 3, respectively) to CHOP in primary therapy of FL. In summary, the relationship established between molecular clearance and minimal residual disease/risk of recurrence may have to be redefined in the times of R. Upcoming large prospective trials will have to elucidate to what degree the clearance of peripheral blood from t(14;18) positive cells can still be regarded as informative regarding absence of minimal residual disease, remission status and/or risk of recurrence.     

CIAPIN1基因在白血病患者骨髓单个核细胞中表达的研究

本研究旨在检测CIAPIN1基因在白血病患者中的表达,探讨其在白血病中的作用。收集112例初治白血病患者的新鲜骨髓,用TRIzoL一步法提取细胞总RNA、合成cDNA、以β-actin为内参,应用实时定量PCR方法检测CIAPIN1mRNA的表达;实验中选择10例正常人的骨髓作为对照。结果表明:骨髓单个核细胞(MNC)中CIAPIN1mRNA在急性髓系白血病(AML)、急性淋巴细胞白血病(ALL)和慢性髓系白血病(CML)患者中的表达均高于正常人(p<0.05);在慢性淋巴细胞白血病(CLL)患者中的表达与正常人相比差异无统计学意义(p>0.05)。结论 :CIAPIN1基因在初治白血病患者MNC中表达升高,其表达上调在白血病的发病中可能有一定的作用。     

食管癌淋巴结转移组织中COL21A1,RUNX2,COAS基因定量检测

目的：探讨3种可能的食管癌转移相关基因COIJ21A1、RUNX2和COAS与食管癌淋巴结转移的相关性。方法：选择4例经病理证实为食管鳞状细胞癌伴淋巴结转移病例，应用实时荧光定量PCR(RQ—PCR)技术测量其淋巴结转移灶中COL21A1、RUNX2和COAS基因的扩增倍数。结果：COL21A1扩增倍数分别为2.50，31.34，0.68，6．45：RUNX2扩增倍数分别为4．44，1．29，0．23，3．25；COAS扩增倍数分别为2．11，10．13，0．44，4．99。3种基因在3／4食管癌转移淋巴结组织中表达增强。结论：COL21A1，RUNX2和COAS基因可能与食管癌的淋巴结转移有关。