Human methionine synthase: cDNA cloning and identification of mutations in patients of the cblG complementation group of folate/cobalamin disorders

Methionine synthase catalyzes the remethylation of homocysteine to methionine in a methylcobalamin-dependent reaction. We used specific regions of homology within the methionine synthase sequences of several lower organisms to clone a human methionine synthase cDNA by a combination of RT-PCR and inverse PCR. The enzyme is 1265 amino acids in length and contains the seven residue structure-based sequence fingerprint identified for cobalamin-containing enzymes. The gene was localized to chromosome 1q43 by the FISH technique. We have identified one missense mutation and a 3 bp deletion in patients of the cblG complementation group of inherited homocysteine/folate disorders by SSCP and sequence analysis, as well as an amino acid substitution present in high frequency in the general population. We discuss the possibility that a mild deficiency of methionine synthase activity could be associated with mild hyperhomocysteinemia, a risk factor for cardiovascular disease and possibly neural tube defects.      

Immunolocalization of the apoptosis regulating proteins Bcl-2 and Bax in human endometrium and isolated peritoneal fluid macrophages in endometriosis

Endometriosis, a debilitating disease associated with infertility, is characterized by the prolonged presence of ectopic endometrial tissue and the involvement of activated peritoneal fluid macrophages. Apoptosis, which occurs in both endometrium and peritoneal fluid macrophages, is controlled in part by members of the Bcl-2/Bax family of proteins. Here, through immunohistochemical staining, we investigated the Bcl-2/Bax status in endometrium and peritoneal fluid macrophages in endometriosis. Bcl-2/Bax immunoreactivity was found predominantly in the glandular epithelial cells, mainly during the proliferative phase of the menstrual cycle for Bcl-2 but throughout the entire menstrual cycle for Bax. Ectopic endometrium contained a population of Bcl-2 positive. Bax negative tissue macrophages. Fluorescence-activated cell sorting of isolated peritoneal fluid macrophages showed that women with endometriosis had a significantly higher proportion of Bcl-2 positive macrophages than the non- endometriotic group. The proportion of Bax positive peritoneal fluid macrophages was significantly elevated in women without endometriosis. The increased proportion of Bcl-2 positive macrophages found in women with endometriosis may predispose these cells to resist apoptosis. The continued survival of these active cells could have important consequences for the survival and proliferation of the ectopic endometrial tissue.      

How to Increase Reach and Adherence of Web-Based Interventions: A Design Research Viewpoint

Nowadays, technology is increasingly used to increase people’s well-being. For example, many mobile and Web-based apps have been developed that can support people to become mentally fit or to manage their daily diet. However, analyses of current Web-based interventions show that many systems are only used by a specific group of users (eg, women, highly educated), and that even they often do not persist and drop out as the intervention unfolds. In this paper, we assess the impact of design features of Web-based interventions on reach and adherence and conclude that the power that design can have has not been used to its full potential. We propose looking at design research as a source of inspiration for new (to the field) design approaches. The paper goes on to specify and discuss three of these approaches: personalization, ambient information, and use of metaphors. Central to our viewpoint is the role of positive affect triggered by well-designed persuasive features to boost adherence and well-being. Finally, we discuss the future of persuasive eHealth interventions and suggest avenues for follow-up research.     

含钛镍铬烤瓷合金的金瓷结合强度实验

目的:比较中国科学院金属研究所研制的3种成分及含量不同的含钛镍铬(Ni-Cr)烤瓷合金与Vita VMK88瓷的结合性能。方法:实验于2005-05/12在中国医科大学口腔医学院技工室和中国科学院金属研究所完成。采用铸造法制作实验合金试件,3种含钛镍铬烤瓷合金依成分不同分为高钛组(含钛量最高且不含铍的Ni-Cr组),中钛组(含钛量较高且不含铍的Ni-Cr组),低钛组(含钛最少且含铍的Ni-Cr组),以HI BOND非贵金属烤瓷合金为对照组,每组试件8个。在试件中份熔附厚度为2mmVita VMK88瓷,形成4.4mm×3.8mm×2.0mm的瓷块。在万能试验机上采用剪切力试验测试金瓷分离时的载荷。结果:高钛组、中钛组、低钛组的剪切强度值低于对照组[分别为(28.4364±4.7918),(25.5335±3.9612),(24.6663±4.4316),(31.1975±3.7038)MPa],各组的剪切强度均值间进行两两比较(q检验),差异均无显著性意义(P>0.05)。结论:3种含钛镍铬烤瓷合金均与Vita VMK88瓷有良好的结合力。     

外侧半月板切除术后膝关节周围骨密度的改变

目的:观察外侧半月板切除术后膝关节周围骨密度的改变。方法:选择1984/2002年唐山市第二医院骨科、华北煤炭医学院附属医院以及开滦医院施行外侧半月板切除术的患者128例,其中男64例,女64例;年龄14～68岁。实验分组:根据术后时间分为:0～1年组32例,2～4年组32例,5～10年组30例和11～17年组34例,设健侧为对照组。方法及评估:应用双能X射线骨密度仪分别测量膝关节周围6个感兴趣区骨密度(6区:以胫骨上端松质骨区腓骨小头水平面作为参考平面,在胫骨外、内侧分别作0.8cm×0.8cm的感兴趣区,称为R1,R2。以股骨下端股骨髁间切迹水平面作为参考平面,在股骨内外侧髁分别作0.8cm×0.8cm的感兴趣区,称为R3,R4。在胫骨上端密质骨区胫骨平台外、内侧软骨下分别作1.4cm×0.4cm的感兴趣区,称为R5,R6)并计算出外侧、内侧骨密度的变化值,用均数百分数±标准差表示。结果:128例患者全部进入结果分析。①在术后17年内胫骨上端外内侧(R56)密质骨骨密度变化值:0～17年术侧均明显高于健侧[0～1年:(-10.20±13.62)%,(-22.80±13.54)%;2～4年:(-8.87±15.05)%,(-17.82±11.06)%;5～10年:(5.06±18.64)%,(-8.62±12.97)%;11～17年:(6.17±14.56)%,(-7.86±10.73)%,t=2.41～8.95,P<0.05]。②5～17年胫骨上端外内侧(R12)松质骨骨密度变化值:5～17年术侧均明显高于健侧[5～10年:(12.15±22.48)%,(0.40±14.40)%,11～17年:(8.16±13.37)%,(-4.90±10.44)%,t=2.31,2.40,P<0.05]。③11～17年股骨下端近膝关节区外内侧(R43)骨密度变化值:术侧明显高于健侧[(4.43±13.70)%,(-4.68±17.68)%,t=2.17,P<0.05]。结论:外侧半月板切除术后可引起膝关节周围骨密度的改变,发生适应性的骨重构:胫骨上端密质骨区最早发生,胫骨上端松质骨区其后发生,股骨下端近膝关节区最后发生。     

微核实验评价自制烤瓷合金的生物相容性

目的：通过微核实验检测自制Ni-Cr-Ti烤瓷合金对小鼠的致畸作用,评价其生物相容性. 方法：实验于2006-04/08在中国医科大学中心实验室完成.①实验分组：选择N1H纯系小鼠80只,按随机数字表法分为8组,每组10只：Ni-Cr-Ti合金组（剂量分别为0.02,0.002,0.000 2 mL/kg）、纯钛浸提液组（剂量分别为0.02,0.002,0.000 2 mL/kg）,阴性对照组（生理盐水）,阳性对照组（注射用环磷酰胺,75 mg/kg）.②实验方法：按50 mL/kg口服同剂量的浸提液,各组均于一次给药后24 h,麻醉后处死小鼠,取小鼠骨髓细胞.③实验评估：光镜下计数1 000个嗜多染红细胞的微核细胞数,并计数嗜多染红细胞与正红细胞的比值,即P/N比值.比值≥1,属于正常,比值＜1,则可能受试物对骨髓细胞产生毒性. 结果：Ni-Cr-Ti合金组、纯钛浸提液组每1 000个嗜多染红细胞的微核细胞数与阴性对照组比较,差异无显著性意义（P＞0.05）,即Ni-Cr-Ti合金、纯钛浸提液对骨髓细胞无毒性.阳性对照组每1 000个嗜多染红细胞的微核细胞数高于Ni-Cr-Ti合金组、纯钛浸提液组,差异有显著性意义（P＜0.01）,对骨髓有毒性作用.Ni-Cr-Ti合金组、纯钛浸提液组P/N比值均≥1. 结论：自制Ni-Cr-Ti合金对小鼠无致畸作用,有较良好的生物相容性.     

Thalidomide as salvage therapy for chronic graft-versus-host disease

Thalidomide has been reported to be an effective agent for treatment of chronic graft-versus-host disease (CGVHD). To determine the efficacy of this agent in patients with refractory CGVHD a total of 80 patients who failed to respond to prednisone (PSE) or PSE and cyclosporine (CSA) were treated with thalidomide. Sixteen patients (20%) had a sustained response, 9 with a complete remission and 7 with a partial response. Twenty-nine patients (36%) had thalidomide discontinued because of side effects, which included sedation, constipation, neuritis, skin rash, and neutropenia. Side effects were reversible with drug discontinuation except for mild residual neuritis in one case. Rashes and neutropenia have not previously been reported as thalidomide side effects when used for CGVHD treatment. We conclude thalidomide is immunosuppressive and active in the treatment of CGVHD. A high incidence of reversible side effects limited dose intensity and reduced the number of patients who could benefit from treatment.     

肝型丙酮酸激酶启动子与人胰岛素基因逆转录病毒介导感染pT67、HepG2细胞的蛋白质表达

目的检测新构建肝型丙酮酸激酶启动子（LPKp）与人胰岛素基因（hInsg）逆转录病毒表达载体（pM54,LPKp-hInsg）在pT67、HepG2细胞的表达情况,为基因治疗或基因结合干细胞治疗糖尿病寻找新的优化生物载体。方法（1）限制酶切父、母本质粒p54、pMDN-SIN,取相关片段构建含人Ins基因一逆转录病毒载体pM54（pMDN-SIN＋p54;LPKp-hInsg）;pM54扩增、纯化、酶切鉴定;（2）脂质体FuGENE6转染pM54质粒进入pT67、PhoenixE和3T3细胞系,同时转染pCMVβGal和父本质粒p54做对照;（3）制备转染后细胞培养拟逆转录病毒上清液;（4）用转染后细胞培养上清液旋转感染pT67、HepG2细胞;（5）ELISA法检测转染、感染后细胞培养上清液Ins含量。结果酶切鉴定质粒与构建预期相符;ELISA法检测出转染、感染上清液中均含较高水平Ins。对照结果均为阴性。结论LPKp-hlnsg基因逆转录病毒表达载体pM54构建成功。人胰岛素基因逆转录病毒介导旋转感染pT67等细胞,目的蛋白Ins获得了预期的表达。实验为基因治疗或基因结合于细胞治疗糖尿病的进一步相关研究奠定了基础。