人NDRG1的GST融合表达、纯化及相互作用蛋白检测

目的: 为了更好的了解NDRG1的功能,通过pull-down技术寻找与NDRG1相互作用的蛋白. 方法: 从已构建好的NDRG1-pPROEXHTb重组载体中酶切下NDRG1 cDNA,插入融合表达载体pGEX-4T-1,序列测定后,转化DH-5α感受态细胞,在含氨苄青霉素的LB中高效表达. 表达的GST-NDRG1经过谷胱甘肽琼脂糖-4B亲和层析纯化,并与高表达细胞系HHCC孵育,通过SDS-PAGE检测和NDRG1相互作用的蛋白. 结果: 纯化蛋白和HHCC孵育后,在Mr 4.0×104处出现新增条带,应是与NDRG1相互作用的未知蛋白. 结论: 在肿瘤细胞HHCC中存在与NDRG1相互作用的蛋白.     

CD25 Is a Novel Marker of Hepatic Bile Canaliculus

Objective. Although many antigens have been investigated, the method for the bile canaliculus staining using optical microscopy needs to be improved. The aim of the present study was to assess the expression pattern of a candidate marker, CD25, in normal and diseased liver tissue. Methods. Immunohistochemistry, immunofluorescence, and immune electron microscopy assays were performed with 41 liver sections and 2 different anti-CD25 monoclonal antibodies. A polyclonal antibody against carcinoembryonic antigen (CEA) was also used to stain bile canaliculus as a control. CD25 expression levels in normal and diseased liver tissue were also determined. Results. CD25 was predominantly localized at the bile canaliculus of adult and infantile liver, evidenced by both immunohistochemistry and immunofluorescence assays. The electron microscopy assay showed that there were obvious amorphous electron-dense deposits at the bile canaliculus. In contrast, the CEA-positive area included bile canaliculus as well as basolateral aspects of hepatocytes. CD25 expression levels did not differ significantly among different disease states. Conclusion. This study provides the first evidence that CD25 is a novel marker of bile canaliculus. Characteristics of CD25 expression may shed light on immunohistochemistry and immunofluorescence analysis of bile canaliculus in both basic and clinical hepatic investigations.     

Voltage- and calcium-dependent gating of TMEM16A/Ano1 chloride channels are physically coupled by the first intracellular loop

Ca(2+)-activated Cl(-) channels (CaCCs) are exceptionally well adapted to subserve diverse physiological roles, from epithelial fluid transport to sensory transduction, because their gating is cooperatively controlled by the interplay between ionotropic and metabotropic signals. A molecular understanding of the dual regulation of CaCCs by voltage and Ca(2+) has recently become possible with the discovery that Ano1 (TMEM16a) is an essential subunit of CaCCs. Ano1 can be gated by Ca(2+) or by voltage in the absence of Ca(2+), but Ca(2+)- and voltage-dependent gating are very closely coupled. Here we identify a region in the first intracellular loop that is crucial for both Ca(2+) and voltage sensing. Deleting (448)EAVK in the first intracellular loop dramatically decreases apparent Ca(2+) affinity. In contrast, mutating the adjacent amino acids (444)EEEE abolishes intrinsic voltage dependence without altering the apparent Ca(2+)affinity. Voltage-dependent gating of Ano1 measured in the presence of intracellular Ca(2+) was facilitated by anions with high permeability or by an increase in [Cl(-)](e). Our data show that the transition between closed and open states is governed by Ca(2+) in a voltage-dependent manner and suggest that anions allosterically modulate Ca(2+)-binding affinity. This mechanism provides a unified explanation of CaCC channel gating by voltage and ligand that has long been enigmatic.     

石河子市社区护理质量管理中存在的问题及思考

有效的护理管理对社区护理质量起到促进作用,要加强有效的人事、质量和医院感染管理,完善各项护理规章制度,提高社区护理质量。     

MicroRNA-27a/b regulates cellular cholesterol efflux,influx and esterification/hydrolysis in THP-1 macrophages

Rationale

Macrophage cholesterol homeostasis maintenance is the result of a balance between influx, endogenous synthesis, esterification/hydrolysis and efflux. Excessive accumulation of cholesterol leads to foam cell formation, which is the major pathology of atherosclerosis. Previous studies have shown that miR-27 (miR-27a and miR-27b) may play a key role in the progression of atherosclerosis.

Objective

We set out to investigate the molecular mechanisms of miR-27a/b in intracellular cholesterol homeostasis.

Methods and results

In the present study, our results have shown that the miR-27 family is highly conserved during evolution, present in mammals and directly targets the 3′ UTR of ABCA1, LPL, and ACAT1. apoA1, ABCG1 and SR-B1 lacking miR-27 bind sites should not be influenced by miR-27 directly. miR-27a and miR-27b directly regulated the expression of endogenous ABCA1 in different cells. Treatment with miR-27a and miR-27b mimics reduced apoA1-mediated cholesterol efflux by 33.08% and 44.61% in THP-1 cells, respectively. miR-27a/b also regulated HDL-mediated cholesterol efflux in THP-1 macrophages and affected the expression of apoA1 in HepG2 cells. However, miR-27a/b had no effect on total cellular cholesterol accumulation, but regulated the levels of cellular free cholesterol and cholesterol ester. We further found that miR-27a/b regulated the expression of LPL and CD36, and then affected the ability of THP-1 macrophages to uptake Dil-oxLDL. Finally, we identified that miR-27a/b regulated cholesterol ester formation by targeting ACAT1 in THP-1 macrophages.

Conclusion

These findings indicate that miR-27a/b affects the efflux, influx, esterification and hydrolysis of cellular cholesterol by regulating the expression of ABCA1, apoA1, LPL, CD36 and ACAT1.     

铜陵县晚期血吸虫病外科治疗效果评价

目的评价铜陵县2005-2007年外科救治晚期血吸虫病人的效果。方法收集铜陵县人民医院外科2005-2007年晚期血吸虫病住院病例资料,进行统计分析。结果共调查72例住院病例,其中2005年15例,2006年19例,2007年38例。男性37例,女性35例。患者年龄最大79岁,最小22岁,平均49.33岁。巨脾型68例,其中腹水合并巨脾型13例;结肠增厚型4例。66例手术治疗患者痊愈出院,2例患者症状明显改善,4例保守治疗患者疗效较差。8例外科治疗晚期血吸虫病患者医药费全部由国家晚期血吸虫病专项救助资金报销,64例承担一部分医疗费用。结论外科治疗晚期血吸虫病效果显著,患者治疗依从性高。     

混合式学习在针灸实训中的设计应用和对照研究

目的　探讨混合式学习在刺法灸法学“毫针刺法”教学中的应用效果。方法　将针灸推拿专业本科二年级学生按照班级分为两组,实验组38人,对照组34人,分别采用混合式学习和传统方式进行“毫针刺法”授课。“混合式学习”为线上微信社区操作练习打卡与线下课堂学习相结合。比较两组学生的课后测试成绩,同时对实验组学生进行问卷调查,以评价混合式学习效果。采用SPSS 19.0进行统计分析,两组数据比较行t检验。结果　实验组“毫针刺法”平均成绩为（4.73±0.15）分,高于对照组的（4.27±0.46）分,差异有统计学意义（t=5.588,P=0.000）。问卷结果显示,88%（22/25）的学生认为,混合式学习对促进针灸操作技能有实效性。结论　混合式学习可以有效提高学生针灸操作技能,并且能够促进学生养成良好学习习惯,使课堂操作练习更有针对性。而更友好的线上学习工具,有待后期研究中进一步挖掘。     

Erythrocytapheresis for chronically transfused children with sickle cell disease: An effective method for maintaining a low hemoglobin S level and reducing iron overload

Cerebrovascular accident (CVA) is a major complication of sickle cell disease during childhood. Long-term transfusion reduces the hemoglobin S level and generally prevents recurrent stroke, but it also results in progressive iron overload that requires regular chelation therapy. Erythrocytapheresis offers an alternative approach aimed at reducing the iron accumulation. We reviewed the results of erythrocytapheresis in eight sickle cell patients (mean age of 12.1 years) at high risk for a first or recurrent stroke. They were maintained at the standard pre-transfusion hemoglobin S (Hb S) level of 30%. Over an average of 9 months of erythrocytapheresis, none of the patients developed complications related to the procedure or to the increased blood use. Ferritin levels decreased by a mean of 26.5% in all patients. When evaluating the ferritin level in five patients, who remained on chelation therapy with deferoxamine (DFO), the level dropped by a mean of 32%. The levels remained stable in the three patients who were not on DFO. The procedure is safe and effective in reducing iron overload and can obviate the need for chelation therapy, even when the target Hb S is maintained at the standard 30% range. J. Clin. Apheresis 14:122–125, 1999. © 1999 Wiley-Liss, Inc.