蛋白质含量测定方法对乌司他丁比活的影响

目的考察2种不同蛋白质含量测定方法对乌司他丁比活的影响。方法分别采用凯氏定氮法和福林酚法对乌司他丁原料药中的蛋白质含量进行测定,再结合效价计算比活。结果由凯氏定氮法得到的比活分别为4 035、3 933和4 010mg·mg-1;由《日本药局方》福林酚法得到的比活分别为3 472、3 360和3 423 mg·mg-1;由《中国药典》2010年版福林酚法得到的比活分别为3 621、3 503和3 570 mg·mg-1。结论由《日本药局方》和《中国药典》福林酚法测得的蛋白质含量结果差异较小,但与凯氏定氮法测得的蛋白质含量差异较大。     

三峡库区血吸虫病潜在流行区安全饮水及无害化厕所使用现状

选取三峡库区血吸虫病潜在流行区的1 386名居民进行问卷调查,结果表明这些地区无害化厕所使用和安全用水情况不容乐观,存在感染血吸虫的潜在危险。     

An engineered TIMP2-based and enediyne-integrated fusion protein for targeting MMP-14 shows potent antitumor efficacy

Recent studies have shown that MMP-14 is highly expressed in a panel of human solid tumors and poses as a potential molecular target for anticancer drugs. Currently, major strategies for targeted therapeutics have mainly focused on the use of antibody or ligand-based agents. For seeking an alternative approach, it is of interest to employ endogenous proteins as drug delivery carriers. Considering the facts that TIMP2, the tissue inhibitor of metalloproteinase 2, shows specific interaction with MMP-14 and that Lidamycin (LDM), an extremely potent cytotoxic antitumor antibiotic, consists of an apoprotein (LDP) and a highly active enediyne (AE); we designed and prepared a TIMP2-based and enediyne-integrated fusion protein LDP(AE)-TIMP2 by DNA recombination and molecular reconstitution consecutively. Furthermore, the MMP-14 binding attributes of the active fusion protein were determined and its therapeutic efficacy against human esophageal carcinoma KYSE150 xenograft and human fibrosarcoma HT1080 xenograft models in nude mice was investigated. It is suggested that TIMP2, the endogenous and MMP-14 binding protein, might serve as a guided carrier for targeted therapeutics.     

Phosphorylation and inactivation of PTEN at residues Ser380/Thr382/383 induced by Helicobacter pylori promotes gastric epithelial cell survival through PI3K/Akt pathway

Phosphorylation of PTEN at residues Ser380/Thr382/383 leads to loss of phosphatase activity and tumor suppressor function. Here, we found that phosphorylation of PTEN at residues Ser380/Thr382/383 was increased with gastric carcinogenesis, and more importantly, Helicobacter pylori was a trigger of this modification in chronic non-atrophic gastritis. H. pylori could phosphorylate and inactivate PTEN in vivo and in vitro, resulting in survival of gastric epithelial cells. Furthermore, stable expression of dominant-negative mutant PTEN or inhibition of Akt prevented the enhanced survival induced by H. pylori. These results indicate that PTEN phosphorylation at residues Ser380/Thr382/383 is a novel mechanism of PTEN inactivation in gastric carcinogenesis, and H. pylori triggers this modification, resulting in activation of the PI3K/Akt pathway and promotion of cell survival.     

Chemokines CCL2, 3, 14 stimulate macrophage bone marrow homing,proliferation, and polarization in multiple myeloma

We previously showed that macrophages (MΦs) infiltrate the bone marrow (BM) of patients with myeloma and may play a role in drug resistance. This study analyzed chemokines expressed by myeloma BM that are responsible for recruiting monocytes to the tumor bed. We found that chemokines CCL3, CCL14, and CCL2 were highly expressed by myeloma and BM cells, and the levels of CCL14 and CCL3 in myeloma BM positively correlated with the percentage of BM-infiltrating MΦs. In vitro, these chemokines were responsible for chemoattracting human monocytes to tumor sites and in vivo for MΦ infiltration into myeloma-bearing BM in the 5TGM1 mouse model. Surprisingly, we also found that these chemokines stimulated MΦ in vitro proliferation induced by myeloma cells and in vivo in a human myeloma xenograft SCID mouse model. The chemokines also activated normal MΦ polarization and differentiation into myeloma-associated MΦs. Western blot analysis revealed that these chemokines promoted growth and survival signaling in MΦs via activating the PI3K/Akt and ERK MAPK pathways and c-myc expression. Thus, this study provides novel insight into the mechanism of MΦ infiltration of BM and also potential targets for improving the efficacy of chemotherapy in myeloma.     

Enolase-1 is a therapeutic target in endometrial carcinoma

ENO1 plays a paradoxical role in driving the pathogenesis of tumors. However, the clinical significance of ENO1 expression remains unclear and its function and modulatory mechanisms have never been reported in endometrial carcinoma (EC). In this study, ENO1 silencing significantly reduced cell glycolysis, proliferation, migration, and invasion in vitro, as well as tumorigenesis and metastasis in vivo by modulating p85 suppression. This in turn mediated inactivation of PI3K/AKT signaling and its downstream signals including glycolysis, cell cycle progression, and epithelial-mesenchymal transition (EMT)-associated genes. These effects on glycolysis and cell growth were not observed after ENO1 suppression in normal human endometrial epithelial cells (HEEC). Knocking down ENO1 could significantly enhance the sensitivity of EC cells to cisplatin (DDP) and markedly inhibited the growth of EC xenografts in vivo. In clinical samples, EC tissues exhibited higher expression levels of ENO1 mRNA and protein compared with normal endometrium tissues. Patients with higher ENO1 expression had a markedly shorter overall survival than patients with low ENO1 expression. We conclude that ENO1 favors carcinogenesis, representing a potential target for gene-based therapy.     

注意力缺陷障碍伴多动儿童个性和行为特点与其家庭环境

目的:分析注意力缺陷障碍伴多动儿童的行为和个性特点及其与家庭环境的关系。方法:于2004/05按照DSM-Ⅳ的注意力缺陷障碍伴多动诊断标准调查长沙市某学校小学三年级到初中一年级共1320名学生,筛查出由儿童精神病学主治医师组确诊的51名注意力缺陷障碍伴多动儿童作为注,对照组为其同班级、同性别、同年龄段(相差<1岁)的正常儿童51名。采用Achenbach儿童行为量表和艾森克儿童个性问卷调查儿童的行为与个性特点,采用家庭环境量表中文版调查儿童家庭环境特征。Achenbach儿童行为量表分为社会能力和行为问题两部分。社会能力由活动情况、社交情况、学校情况3个分量表组成;行为问题分为退缩、躯体主诉、焦虑抑郁、社交问题、思维问题、注意问题、违纪行为、攻击性行为8个分量表,并分别计算社会能力和行为问题总分。其中退缩、躯体主诉、焦虑抑郁、注意问题4个分量表构成了内化性行为问题;违纪行为、攻击性行为两个分量表构成外化性行为问题。艾森克儿童个性问卷有内外倾向、情绪稳定性、精神质和掩饰倾向4个分量表。采用家庭环境量表中文版,包括亲密度、情感表达、矛盾性、独立性、成功性、知识性、娱乐性、道德宗教观、组织性、控制性等10个维度。Achenbach儿童行为量表和家庭环境量表由同学带回家由父母填写,1周后收回,艾森克儿童个性问卷由儿童仔细填写。结果:102名儿童完成全部测评,进入结果分析。①注意力缺陷障碍伴多动组儿童社交情况、学校情况、焦虑抑郁、社交、思维及注意问题评分和社会能力总分和行为总分明显高于对照组(t=-3.966～8.253,P<0.001)。②注意力缺陷障碍伴多动儿童个性在内外倾向得分较对照组低(34.61±5.66,42.31±6.42,t=-6.429,P<0.001),而情绪稳定性及精神质得分较对照组高(34.31±6.29,30.90±5.11,t=3.008,P<0.05);(27.27±3.19,22.88±4.02,t=6.111,P<0.001)。③注意力缺陷障碍伴多动儿童的家庭环境中其亲密度程度、情感表达、独立性、成功性、组织性得分低于正常组(t=-7.801～-2.139,P<0.05或0.001),而矛盾性、控制性得分高于正常组(t=3.099,3.246,P<0.05)。④注意力缺陷障碍伴多动儿童的行为和个性特征与家庭环境无明显相关,取Achenbach儿童行为量表中的思维问题、社交问题、注意问题、内外向性行为问题与家庭环境各因子分做典型相关分析,并得到第一典型相关系数(λ1=0.679,P>0.05);取艾森克儿童个性问卷内外倾向、情绪稳定性和精神质3个维度与家庭环境各因子分做典型相关分析,求得典型相关系数(λ1=0.610,P>0.05)。结论:注意力缺陷障碍伴多动儿童具有突出的行为问题和不同程度社会功能损害。家庭环境对其未产生明显的影响。与正常儿童相比,注意力缺陷障碍伴多动儿童呈现了注意缺陷、攻击和违纪行为的独特个性特征和异常的家庭结构。     

釉基质蛋白对牙周膜成纤维细胞合成蛋白的影响

目的:检测釉基质蛋白作用下牙周膜成纤维细胞合成蛋白的变化,分析釉基质蛋白促进牙周组织再生的作用。方法:实验于2004-04/08在第四军医大学口腔医院组织工程实验室完成。选取11～14岁儿童因正畸需要拔除的无牙周病及龋病的健康新鲜前磨牙,锐利刀片刮除牙根颈部1/3牙龈及牙周膜,采用全牙胶原酶消化法培养获得牙周膜成纤维细胞,经免疫细胞化学检测证实为外胚间充质细胞。取第3代细胞用于实验,分为2组:对照组为含10mL/L新生牛血清的低糖DMEM培养液,实验组为100mg/L釉基质蛋白,用含10ml/L新生牛血清的低糖DMEM培养液配制。将第3代人牙周膜成纤维细胞制备成密度为1×104/mL的细胞悬液,接种入预先放置有细胞爬片的6孔板中,24h后按实验分组更换诱导液。逐日倒置显微镜下观察细胞形态。第3,7天利用免疫组化方法和图像分析技术检测两种细胞骨涎蛋白、骨桥蛋白、骨粘连蛋白、牙骨质附着蛋白的表达。结果:①细胞形态学观察:加入诱导因子初期细胞形态无明显变化,随时间延长,实验组细胞突起逐渐变短,与对照组相比有明显差异。②免疫细胞化学检测结果:对照组骨涎蛋白、骨桥蛋白呈弱阳性到阳性表达,实验组与对照组相比,骨涎蛋白、骨桥蛋白均呈现不同程度胞浆着色加深,与作用时间成正比。对照组细胞骨粘连蛋白基本为阴性表达,而实验组细胞第3天时即检测到了骨粘连蛋白的表达,且随时间的延长染色加深。未经釉基质蛋白处理的牙骨质附着蛋白抗体为阴性表达,经过釉基质蛋白作用第3,7天均检测到牙骨质附着蛋白抗体的弱阳性表达,胞浆着色呈淡棕黄色。设立的空白对照和阴性对照均未见着色。③釉基质蛋白对人牙周膜成纤维细胞合成骨涎蛋白、骨桥蛋白、骨粘连蛋白的影响:与对照组比较,实验组第3,7天骨涎蛋白和骨桥蛋白均明显上调(184.31±5.67,168.20±6.93;181.42±4.99,163.91±5.86;213.02±5.40,194.81±5.06;211.12±5.59,182.06±6.66;P均<0.01),骨粘连蛋白对照组未表达,实验组第3天时检测到表达;实验组第7天骨涎蛋白、骨桥蛋白、骨粘连蛋白上调幅度均明显高于第3天(163.91±5.86,168.20±6.93,P<0.05;182.06±6.66,194.81±5.06,P<0.01;196.73±5.47,204.67±5.12,P<0.01)。结论:釉基质蛋白对人牙周膜成纤维细胞具有上调骨涎蛋白、骨桥蛋白、骨粘连蛋白的作用,随时间延长这种促进作用愈明显;并能诱导细胞表达牙骨质附着蛋白。釉基质蛋白在一定浓度下可使人牙周膜成纤维细胞向成骨、成牙骨质样细胞分化。