黄芩苷下调NF-κB表达抑制高糖环境下肾小球系膜细胞增殖的作用

目的研究黄芩苷对高糖环境下肾小球系膜细胞（GMC）增殖的抑制作用及对NF-κB表达的影响。方法将体外培养的大鼠GMC分为六组：低糖组,高糖组,黄芩苷低、中、高剂量组、氯沙坦组,培养24、48、72 h。实验结束后根据MTT法检测结果,收集细胞并提取核蛋白,用Western blot方法检测各组NF-κB表达情况。结果高糖组在24、48、72 h时间段的吸光值均较低糖组增高,且随培养时间延长而增高明显（P〈0.05）;黄芩苷低、中、高剂量组,氯沙坦组在24 h时间段吸光值均较高糖组高,48、72 h时间段较高糖组低（P〈0.05）。Western blot结果显示,高糖组NF-κB表达量较低糖组增加。而用黄芩苷干预后,GMC中NF-κB表达量较高糖组减少,且随着浓度增加,表达量逐渐减少。结论高糖环境下体外培养的GMC存在异常增殖现象,黄芩苷能抑制高糖诱导的GMC异常增殖,且呈时间、剂量依赖关系,其机制可能与黄芩苷下调NF-κB表达有关。     

Estimation of aneuploidy for chromosomes 3, 7, 16, X and Y in spermatozoa from 10 normospermic men using fluorescence in-situ hybridization

Fluorescence in-situ hybridization (FISH) is a fast and efficient method of estimating aneuploidy in human spermatozoa. In this study, we have estimated baseline disomy frequencies in spermatozoa from a group of 10 normospermic men, using stringent scoring criteria. A triple- probe FISH procedure was used for chromosomes 3, X and Y, while a double-probe FISH method was used for chromosomes 7 and 16. A total of 101273 spermatozoa were scored for chromosomes 3, X and Y, resulting in 97.83% haploidy (3X or 3Y), 0.39% disomy (33X, 33Y, 3XX, 3YY or 3XY) and 0.35% diploidy (33XX, 33YY or 33XY). A total of 100760 spermatozoa were scored for chromosomes 7 and 16, giving 98.9% haploidy (716), 0.11% disomy (7716 or 71616) and 0.27% diploidy (771616). Disomy frequencies for individual chromosomes differed (chromosome 3, 0.20%; chromosome 7, 0.05%, chromosome 16, 0.06%; X + Y, 0.19%). The frequency of disomy 3 was significantly higher than disomy 7 (P = 0.019) and disomy 16 (P = 0.022), while the frequency of sex chromosome disomy was significantly higher than disomy 7 (P = 0.0058) and disomy 16 (P = 0.0067), but not disomy 3 (P = 0.73). The disomy and diploidy (0.27- 0.35%) estimates obtained for this normospermic population were generally low and were similar to other recent reports.      

Treatment of acute lymphoblastic leukemia in adults with intensive induction, consolidation, and maintenance chemotherapy

The Southwest Oncology Group conducted a study of acute lymphoblastic leukemia (ALL) in adults over a 5-year period, testing the utility of the L-10M regimen initially described by the group from Memorial Sloan- Kettering Cancer Center. One hundred sixty-eight eligible patients were treated with this intensive combination chemotherapy regimen. One hundred fifteen (68%) achieved complete remission. With the current median follow-up time of 34.5 months, the median durations of remission, relapse-free survival, and overall survival were 22.9, 20.9, and 17.7 months, respectively. Only 35% of the patients over 50 years of age achieved a complete remission. Age was a significant prognostic factor for complete response, survival, relapse-free survival, and remission duration. In addition, a low initial WBC count was found to have a statistically significant association with longer remission duration. Responders between the ages of 20 and 49 years with WBC counts of less than 15,000 appear to have an exceptionally good prognosis.     

脐血间充质干细胞体外培养方法的改良及其成骨分化能力

目的:采用两种改良方法体外分离培养脐血间充质干细胞,并观察其成骨分化能力。方法:实验于2006-05/09在上海交通大学医学院附属第九人民医院骨与关节中心细胞实验室完成。①所用28份脐血标本由复旦大学医学院附属妇产科医院提供,取自足月健康顺产新生儿,产妇和家属均书面同意,实验经医学伦理委员会批准。②采集28份脐血标本,50～90mL/份,枸橼酸抗凝。采集后12h内密度梯度离心法分离出单个核细胞,接种于100mm×20mm培养皿中,细胞浓度为1×1010L-1,置于含体积分数为0.1胎牛血清的α-MEM培养液中原代培养,5～7d后半量换液,后每隔3～4d全量换液一次。③细胞贴壁后,分两组予以改良培养。改良1组10份,当培养皿底圆形巨核细胞融合、梭形成纤维样细胞脱落时将细胞悬液移入新的培养皿中培养;改良2组18份,待培养皿底圆形巨核细胞渐渐占据优势时,将培养基更换为含体积分数为0.15小牛血清的α-MEM,当圆形巨核细胞大部脱落后换回含体积分数为0.1胎牛血清的α-MEM培养基。成纤维样细胞融合至80%～90%时胰酶消化,按1∶2或1∶3传代培养。④显微镜下观察脐血间充质干细胞的形态。取第5代脐血间充质干细胞,采用流式细胞仪测定细胞免疫表型,以碱性磷酸酶法检测成骨分化能力。结果:①28份脐血间充质干细胞中,共20份原代培养中出现贴壁细胞(改良1组6份,改良2组14份),其中13份培养出能融合且可稳定传代的成纤维样细胞(改良1组4份,改良2组9份),成功率46.4%。②原代培养5～7d后贴壁细胞呈梭形成纤维样细胞和圆形巨核细胞。改良1组与2组于原代培养5周可见成纤维样细胞集落,细胞形态与骨髓间充质干细胞相似,呈较均一的长梭形,传至22代形态无明显变化。③可强烈表达CD29、CD105等间充质干细胞表面标志,而不表达CD34、CD45和CD106等造血干细胞和内皮细胞标志。④成骨诱导1周,脐血间充质干细胞可分化为成骨细胞,碱性磷酸酶染色呈阳性。结论:脐血中存在的单个核细胞经过改良培养后,可提高脐血间充质干细胞的培养成功率。脐血间充质干细胞具有与其他来源的单个核细胞类似的表型及成骨分化潜能,且易于体外扩增、传代稳定。     

Percutaneous insertion of the Kimray-Greenfield filter: technical considerations and problems

Kimray-Greenfield filters were inserted percutaneously into the inferior vena cava (IVC) in 57 patients. Thirty-six were placed from the right femoral vein, 14 from the left femoral vein, and seven from the right internal jugular vein. There were no deaths or major complications and only six minor complications. Inferior vena cavography was done before filter insertion in all cases. Cavography is vital to determine feasibility of filter insertion, route of insertion, and filter location; pertinent findings include caval size, presence or absence of clot in the IVC or iliac veins, and position of the renal veins. The guide wire provided with the standard filter introduction set has a tendency to catch on the filter as the wire is withdrawn. A stiff wire with a straight, tapered, floppy tip was substituted. The femoral approach is preferred when it is feasible. Though there was only one known occurrence of femoral vein thrombosis at the filter insertion site, other cases may have occurred and may not have been detected. If the frequency of this complication proves to be significant, the preferred route for filter insertion may have to be reconsidered.     

Expression of CYP1B1 but not CYP1A1 by primary cultured human mammary stromal fibroblasts constitutively and in response to dioxin exposure: role of the Ah receptor

The expression of CYP1B1 in human mammary fibroblasts (HMFs) was characterized as a potential modulator of their individual function as well as effects on adjacent mammary epithelia. We have used these characteristics to explore the diversity of fibroblast cells isolated from reduction mammoplasty patients and from different breast locations in breast cancer patients (tumors, peripheral to tumor and skin). These parameters have also been used to examine differences between two donors. The results have shown that while none of these HMFs expressed a detectable CYP1A1 protein basally or in response to TCDD, they all expressed CYP1B1 constitutively at similar levels (0.5-0.9 pmol/mg microsomal proteins) and they were induced by TCDD (up to 5-fold) consistent with mediation by the Ah receptor (AhR). DMBA metabolism by HMFs exhibited high proportions of 5,6-, 10,11- and 3,4-dihydrodiols, a profile that is typical of human CYP1B1 regioselectivity. RT-PCR followed by Southern blot analyses demonstrated that CYP1B1 mRNA expression in HMFs parallels levels of respective microsomal proteins. The AhR is expressed in these HMFs as two cytosolic forms (approximately 106 and 104 kDa) and a substantial proportion of the 104 kDa form was localized to the nucleus even prior to TCDD treatment. In all HMFs isolated directly from collagenase digested breast tissues the AhR is expressed at levels 10-fold lower than in breast epithelial cells. However, HMFs that were isolated after serial passaging of mammary epithelial cultures had shown much higher levels of the AhR expression and more dramatic TCDD-induced down-regulation (>80% in 24 h) associated with more efficient nuclear translocation. These differences suggested the presence of two functionally distinct subtypes of HMFs: interstitial stromal fibroblasts that are readily released by collagenase digestion of breast tissues, and lobular stromal fibroblasts which are more tightly associated with the breast epithelia.      

Treatment of new onset refractory status epilepticus/febrile infection-related epilepsy syndrome with tocilizumab in a child and a young adult

New onset refractory status epilepticus (NORSE) is a rare and devastating condition occurring in a previously healthy patient. It is called febrile infection-related epilepsy syndrome (FIRES) when preceded by a febrile infection. It often leads to intensive care treatment, including antiseizure drugs in combination with anesthetic agents, and sometimes ketogenic diet. The mortality rate is high, and severe epileptic and neuropsychiatric sequelae are usually observed. Based on the possible role of neuroinflammation, intravenous immunoglobulin, corticosteroids, and immunomodulatory treatment (anti-IL1, IL6) can be added. We describe here a child and a young adult with FIRES, both treated with tocilizumab. We observed a rapid positive response on the status epilepticus and good tolerance, but different neurological outcomes for our two patients. Further prospective studies may be necessary both to confirm the efficacy and the safety of this promising treatment and to optimize the immunomodulatory strategy in FIRES/NORSE.