登革Ⅱ型病毒M株、NGC株E基因部分序列原核蛋白多克隆抗体的制备

目的制备登革Ⅱ型病毒M株、NGC株E基因部分序列原核蛋白的小鼠多克隆抗体并进行鉴定。方法利用SDS-PAGE电泳的方法将纯化后的登革Ⅱ型病毒M株、NGC株E基因部分序列原核表达蛋白浓缩,切割目的条带并研磨制成抗原,皮下多点分次注射BALB/c小鼠,采用Western-Blot、间接免疫荧光法对小鼠血清多克隆抗体进行鉴定。结果经SDS-PAGE电泳成功地将纯化后的登革Ⅱ型病毒M株、NGC株E基因部分序列原核表达蛋白浓缩,并获取了抗登革Ⅱ型病毒M株、NGC株E基因部分序列原核表达蛋白的多克隆抗体。结论本实验中所用到的登革Ⅱ型病毒M株、NGC株E基因部分序列原核蛋白为可溶性抗原,通过切胶研磨制成颗粒性蛋白抗原能获得良好的免疫效果,该方法制备的抗原浓度及纯度高,由此获得了高效价高特异性的多克隆抗体。     

Anisomycin induces glioma cell death via down-regulation of PP2A catalytic subunit in vitro

Aim:

To examine the effects of anisomycin on glioma cells and the related mechanisms in vitro.

Methods:

The U251 and U87 human glioblastoma cell lines were tested. The growth of the cells was analyzed using a CCK-8 cell viability assay. Apoptosis was detected using a flow cytometry assay. The expression of proteins and phosphorylated kinases was detected using Western blotting.

Results:

Treatment of U251 and U87 cells with anisomycin (0.01–8 μmol/L) inhibited the cell growth in time- and concentration-dependent manners (the IC₅₀ values at 48 h were 0.233±0.021 and 0.192±0.018 μmol/L, respectively). Anisomycin (4 μmol/L) caused 21.5%±2.2% and 25.3%±3.1% of apoptosis proportion, respectively, in U251 and U87 cells. In the two cell lines, anisomycin (4 μmol/L) activated p38 MAPK and JNK, and inactivated ERK1/2. However, neither the p38 MAPK inhibitor SB203580 (10 μmol/L) nor the JNK inhibitor SP600125 (10 μmol/L) prevented anisomycin-induced cell death. On the other hand, anisomycin (4 μmol/L) reduced the level of PP2A/C subunit (catalytic subunit) in a time-dependent manner in the two cell lines. Treatment of the two cell lines with the PP2A inhibitor okadaic acid (100 nmol/L) caused marked cell death.

Conclusion:

Anisomycin induces glioma cell death via down-regulation of PP2A catalytic subunit. The regulation of PP2A/C exression by anisomycin provides a clue to further study on its role in glioma therapy.     

Corosolic acid analogue,a natural triterpenoid saponin,induces apoptosis on human hepatocarcinoma cells through mitochondrial pathway in vitro

Context 2a,-3a,-24-Trihydroxyurs-12-en-28-oic acid (TEO, a corosolic acid analogue) is a triterpenoid saponin isolated from Actinidia valvata Dunn (Actinidiaceae), a well-known traditional Chinese medicine.

Objective This study investigated the anti-proliferation and inducing apoptosis effects of TEO in three human hepatocellular carcinoma (HCC) cell lines.

Materials and methods Cytotoxic activity of TEO was determined by the MTT assay at various concentrations from 2.5 to 40?μg/mL in BEL-7402, BEL-7404 and SMMC-7721 cell lines. Cell morphology was assessed by acridine orange/ethidium bromide and 4′-6-diamidino-2-phenylindole dihydrochloride staining and fluorescence microscopy. Cell-cycle distribution and DNA damage were determined by flow cytometry and comet assay. Mitochondrial dysfunction was assessed by JC-1 staining and transmission electron microscopy. Apoptosis changes were explored by Western blot, TNF-α and caspase-3, -8, -9 assays.

Results TEO exhibited inhibition effects on BEL-7402, BEL-7404 and SMMC-7721 cells treated for 24?h, the IC₅₀ values were 34.6, 30.8 and 30.5?μg/mL, respectively. TEO (40?μg/mL)-treated three cell lines increased by more than 21% in the G1 phase and presented the morphological change and DNA damage. TEO also declined the mitochondrial membrane potential and altered mitochondrial ultra-structure. Furthermore, caspase-3, caspase-8, caspase-9 and TNF-α were also activated. Mechanism investigation showed that TEO could decrease anti-apoptotic Bcl-2 protein expression, increase proapoptotic Bax and Bid proteins expressions and increase Bax/Bcl-2 ratio.

Conclusion Our results demonstrate for the first time that TEO inhibited growth of HCC cell lines and induced G1 phase arrest. Moreover, proapoptotic effects of TEO were mediated through the activation of TNF-α, caspases and mitochondrial pathway.     

复方丹参滴丸对高脂血症患者红细胞变形性的影响及其机制分析

目的 观察复方丹参滴丸对高脂血症患者红细胞变形性的影响,并探讨其可能的作用机制.方法 选取高脂血症患者100例,完全随机分为单药组和联合组,各50例.单药组予以瑞舒伐他汀钙10 mg/次,每晚1次口服;联合组在单药组基础上给予复方丹参滴丸10粒/次,3次/d口服.8周后检测红细胞变形性、红细胞膜Na+-K+-ATP酶活性及丙二醛含量的变化.结果 ①单药组治疗前TG、TC、LDL-C、HDL-C分别为(2.62±0.53)、(6.5±1.0)、(4.2±0.5)、(1.2±0.3)mmol/L,治疗后分别为(1.45±0.29)、(4.2±1.0)、(2.9±0.4)、(1.7±0.3)mmol/L,治疗前后差异均有统计学意义(P＜0.01或P＜0.05);联合组治疗前TG、TC、LDL-C、HDL-C分别为(2.51±0.33)、(6.8±1.0)、(4.0±0.5)、(1.1±0.3)mmol/L,治疗后分别为(1.40±0.31)、(4.2±0.9)、(2.8 ±0.5)、(1.7±0.4)mmol/L,治疗前后差异均有统计学意义(P＜0.01或P＜0.05),治疗后组间差异无统计学意义(P＞0.05).②单药组和联合组治疗前红细胞变形指数分别为(56±4)％和(59±6)％,治疗后红细胞变形指数分别为(62±8)％和(69±8)％,组内治疗前后的差异及治疗后组间差异均有统计学意义(均P ＜0.05).③单药组和联合组治疗前红细胞膜Na+-K+-ATP酶活性分别为(0.57±0.06)和(0.61±0.07) μmol/(mg·min),治疗后分别为(0.81±0.09)和(1.05±0.11) μmol/(mg·min),组内治疗前后的差异及治疗后组间差异均有统计学意义(均P＜0.05).④单药组和联合组治疗前血浆丙二醛水平分别为(6.3±0.5)和(6.2±0.7) μmol/L (P ＜0.05),治疗后分别为(5.6±0.6)和(5.1±0.4)μmol/L,组内治疗前后的差异及治疗后组间差异均有统计学意义(均P＜0.05).结论 复方丹参滴丸能有效改善高脂血症患者红细胞变形性,提高红细胞通过微小血管的能力,其机制可能与降低脂质过氧化损伤以及提高红细胞膜Na+-K+-ATP酶活性有关.     

腺垂体功能减退症及其危象62例临床分析

目的探讨腺垂体功能减退症的临床特征及治疗方法。方法对收治的62例腺垂体功能减退症患者的临床资料进行回顾性分析。62例患者中席汉综合征40例,垂体瘤7例,颅咽管术后放疗5例,空泡蝶鞍3例,恶性肿瘤脑转移外放疗后2例,自身免疫性垂体炎1例,垂体瘤术后并发症1例,感染性休克1例,糖尿病1例,特发性1例。结果 62例患者均给予氢化可的松或泼尼松替代治疗,其中48例患者联合左甲状腺素片治疗,15例病情较轻者补充性激素,予雌、孕激素序贯治疗,2例补充重组生长激素。12例垂体危象全部救治成功。62例患者均接受随访,死亡13例,死亡原因:未治垂体危象6例,心肌梗死4例,并发肺癌2例,低血糖昏迷1例。结论腺垂体功能减退症临床表现复杂,应提高临床医生对该病的诊治水平,避免误、漏诊。     

口腔溃疡豚鼠血清中谷胱甘肽过氧化物酶活性的研究

目的：研究口腔溃疡豚鼠血清中谷胱甘肽过氧化物酶（GSH-PX）的活性。方法：用90%苯酚溶液灼烧豚鼠左侧面颊,造成口腔溃疡模型,观察豚鼠口腔溃疡模型的溃疡面积、溃疡程度及溃疡组织病理变化,测定血清中GSH-PX活性。结果：用苯酚溶液造豚鼠口腔溃疡模型成功,d1～d5,模型组与空白组相比,口腔溃疡程度显著,血清中GSH-PX活性显著降低（P<0.01）。口腔溃疡豚鼠随着时间的变化溃疡面积逐渐缩小,血清中GSH-PX活性逐渐增强。结论：血清中GSH-PX对口腔溃疡的自愈有促进作用。     

移植性肿瘤的自然消退与免疫形成的研究

本文对小鼠移植性肿瘤的自然消退与其免疫形成进行观察,结果表明:自然消退的EAC昆明鼠对肿瘤的再次攻击具有强烈的排斥反应。应用化疗药物RS034(Sodium sald.ofbishemisuccinate of 7β-Hydroxycholesterol)治愈的L_(1210) DBA/2小鼠进行肿瘤再次移植时,同样产生对抗作用。而用化疗药物5-Fu治愈的小鼠没有出现免疫排斥反应。     

Preparation and characterisation of the colistin-entrapped liposome driven by electrostatic interaction for intravenous administration

Potential use of liposome for polycationic colistin is hindered by their phospholipid membrane permeability. In this study, liposomes were modified with sodium cholesteryl sulphate (Chol-SO₄^?) for improving the colistin loading by enhancing the colistin-bilayer electrostatic attraction. We have evaluated two liposomes: colistin-entrapped liposome of Chol-SO₄^? (CCL) and coated Chol-SO₄^?/colistin complex liposome (CCCL). In comparison with CCL which formed large aggregates at Chol-SO₄^?/colistin charge ratio below 2:1, CCCL showed a smaller size less dependent on the charge ratio, probably arising from more colistin entrapped on the inner leaflet of bilayer. Both liposomes exhibited significantly increased entrapment efficiency as compared with the liposome without Chol-SO4^?. But colistin released upon dilution, implying free transfer of colistin through bilayers. Pharmacokinetics results showed the approximately four-fold increase in the plasma AUC_0–8?h for CCCL and CCL as compared with colistin solution, showing potential benefit for infectious target localisation by prolonging the systemic circulation of colistin.     

正常中国汉人红细胞中儿茶酚氧位甲基转移酶的活性测定

目的测定正常中国人群红细胞儿茶酚氧位甲基转移酶(COMT)活性浓度值,为研究COMT活性变化在疾病诊断中的应用提供依据。方法采用高效液相色谱法测定279名正常健康人(其中男性134例,女性145例)红细胞中COMT的活性浓度,用SPSS统计软件对性别间测定结果进行检验比较。结果男性和女性红细胞中COMT活性浓度分别为:(16.5±7.4)、(15.5±5.3)nmol.mL RBC-1.h-1。活性频率分布男性在6.0~37.4nmol.mL RBC-1.h-1,女性为5.4~37.2 nmol.mL RBC-1.h-1,男女之间COMT活性浓度的差异无统计学意义(P>0.05)。结论本法灵敏,简便,准确,重复性好,且测定结果与国外一些报道一致,适用于临床应用测定研究。本结果可作为中国健康人红细胞COMT活性正常值及今后研究的重要参考数据。     

A new meroditerpenoid from Mayodendron igeum

A new meroditerpenoid, igeumone (1), together with 18 known compounds (2-19), were isolated from ethanolic extract of the bark of Mayodendron igeum. Their structures were determined by analysis of spectral data or comparison with authentic samples. X-ray crystallographic analysis was employed to unambiguously determine the structure of 1.