黑素瘤抑制蛋白在转基因小鼠胚胎乳腺组织中的表达

目的：检测黑素瘤抑制蛋白（ＭＩＡ／ＣＤ－ＲＡＰ）在小鼠胚胎乳腺组织中的表达。方法：剖腹分离出１０．５～１６．５ ｄ 的ＭＩＡ／ＣＤ－ＲＡＰ启动子－半乳糖苷酶（２．２ｌａｃＺ）转基因小鼠胚胎，利用Ｘ－ｇａｌ全胚胎染色观察转基因的组织和细胞染色，并以免疫组织化学染色检测内源性ＭＩＡ／ＣＤ－ＲＡＰ蛋白在胚胎乳腺组织中的表达。结果：ＭＩＡ／ＣＤ－ＲＡＰ在孕期１１．５ ｄ 的小鼠胚胎乳腺上皮细胞中开始表达，至１３．５ ｄ 表达明显增强，而后逐渐减弱，至１６．５ ｄ 消失。结论：ＭＩＡ／ＣＤ－ＲＡＰ可能在乳腺的早期发育和乳腺癌发生中起重要作用。

黑素瘤抑制蛋白启动子在小鼠体内的表达活性

目的：观察黑素瘤抑制蛋白（ＭＩＡ／ＣＤ－ＲＡＰ）启动子在小鼠体内的表达活性。方法：利用ＰＣＲ技术扩增２．２ ｋｂ 的ＭＩＡ／ＣＤ－ＲＡＰ启动子，分子克隆技术构建ＭＩＡ 启动子－半乳糖苷酶（２．２ｌａｃＺ）载体，显微注射法将纯化的２．２ｌａｃＺＤＮＡ 注入来自Ｂ６ＳＪＬ杂交小鼠受精卵的原核以制备转基因小鼠。提取幼鼠尾ＤＮＡ，用ｌａｃＺ特异性引物筛选阳性转基因小鼠。结果：８只首建小鼠染色体基因组整合有２．２ｌａｃＺ融合基因。Ｘ－ｇａｌ全胚胎染色发现其中３ 只转基因小鼠在软骨和乳腺组织中均可见转基因的表达。结论：２．２ ｋｂ

黑素瘤抑制性活性因子在黑素瘤不同阶段及基底细胞癌中的表达

目的:探讨黑素瘤抑制性活性因子(MIA)在黑素瘤及基底细胞癌中的表达及其作用。方法:应用SP免疫组化技术检测38份黑素瘤石蜡标本、35份基底细胞癌石蜡标本以及32份色素痣石蜡标本中MIA的表达水平。结果:MIA在所有色素痣以及基底细胞癌中均呈阴性表达,而在原位黑素瘤、侵袭性黑素瘤、有淋巴结转移的黑素瘤、无淋巴结转移的黑素瘤阳性表达率分别为21.4%、91.6%、94.1%和42.8%。结论:MIA在黑素瘤的发生发展中起重要作用,MIA有可能成为临床诊断、治疗黑素瘤的靶点。

黑素瘤抑制性活性蛋白及基质金属蛋白酶-9在恶性黑素瘤的表达

目的:探讨黑素瘤抑制性活性蛋白(MIA)及基质金属蛋白酶-9(MMP-9)在恶性黑素瘤中的表达以及作用。方法:应用SP免疫组化技术对38例恶性黑素瘤石蜡标本以及32例色素痣石蜡标本检测MIA及MMP-9表达水平。结果:MIA在所有色素痣中表达阴性,而在原位黑素瘤、侵袭性黑素瘤、有淋巴结转移恶性黑素瘤、无淋巴结转移恶性黑素瘤阳性表达率分别为21.4%,91.6%、94.1%、42.8%,MMP-9在恶性黑素瘤阳性表达率为71.1%,MIA的表达与MMP-9的表达呈正相关。结论:MIA在恶性黑素瘤的发生发展中起着重要作用,这种作用可能是通过上调MMP-9的表达来实现的。MIA有可能成为临床诊断、治疗恶性黑素瘤的有力靶点。

黑素瘤抑制蛋白2和VEGF在肝癌中的表达及其临床意义

目的研究黑素瘤抑制蛋白2(MIA2)和血管内皮生长因子(VEGF)在肝癌中的表达及其与肝癌临床病理特征之间的关系。方法采用免疫组化SP法检测36例肝癌手术切除标本的MIA2及VEGF的表达情况。结果 36例HCC中MIA2、VEGF阳性表达分别为41.7%和77.8%;MIA2和VEGF表达有相关性(P<0.05)。29例癌旁组织中MIA2和VEGF阳性表达分别为93.1%和34.5%。肝癌组织和癌旁组织MIA2和VEGF表达差异有统计学意义(P<0.01)。结论 MIA2在肝癌组织中呈低表达,且与肝癌的TNM分期、病理分级及肿瘤转移有关。MIA2和VEGF的表达呈负相关。

黑素瘤抑制活性蛋白作为血清标志物对恶性黑素瘤的诊断价值

恶性黑素瘤(malignant melanoma,MM)是起源于神经嵴黑素细胞的高度恶性肿瘤。由于MM具有高度侵袭性且转移后预后极差,寻找一个可以提示肿瘤进展并评估预后的诊断指标显得极为重要。黑素瘤抑制活性蛋白(melanoma inhibitory activity protein,MIA)是一种在MM患者体内特异性高表达的小分子蛋白,在转移性MM患者血清水平增高尤为明显。研究证实,MIA参与了MM的转移和免疫抑制,故检测MIA水平对监测原发灶切除后肿瘤的复发和转移十分必要,同时解决了传统复查方式的辐射暴露和高昂检测费用等问题。该文综述了近年来MIA的分子机制研究、MIA的检测方法及是否可作为预后指标,还综述了与其他血清标志物如S-100β蛋白、乳酸脱氢酶(lactic dehydrogenase,LDH)结合,评估MM结局,认为MIA可成为评估MM进展及预后的指标。

人黑素瘤细胞A375 MIA/CD-RAP基因的克隆及其鉴定

目的为了研究人黑素瘤M IA/CD-RAP基因的生物学功能,克隆M IA/CD-RAP的cDNA,构建表达载体后,对其进行酶切和DNA测序鉴定。方法用RT-PCR从培养的人黑素瘤细胞A375中扩增M IA/CD-RAP cDNA,将其克隆到载体pcDNA3.1-MycH is上,经酶切及PCR扩增鉴定和测序。结果酶切电泳和DNA测序结果表明,成功地克隆了人黑素瘤M IA/CD-RAP cDNA。结论人黑素瘤M IA/CD-RAP cDNA的克隆,可为进一步研究其生物学功能奠定基础。

血清MIA检测在恶性黑素瘤诊断中的临床价值

目的 :探讨血清黑素瘤抑制蛋白 (melonom a- inhibiting activity protein,MIA)在恶性黑素瘤诊断中的价值。方法 :采用 EL ISA方法 ,检测不同临床分期的黑素瘤及其他恶性肿瘤患者 ,以及正常人血清 MIA浓度。 结果 :正常成人 (n=2 0 )和肺癌 (n=10 )、肝癌 (n=12 )、结肠癌 (n=12 )患者血清 MIA浓度全部低于正常 (6 .5 ng/m l)水平 ,黑素瘤患者血清 MIA浓度明显升高 ,阳性率达 77.3% (17/2 2 ) ,其中 , 期患者阳性率达 10 0 % (15 /15 )。乳腺癌患者阳性率 2 6 .7% (4/15 )。 结论 :血清MIA水平是临床诊断黑素瘤的一个较好指标。

人黑素瘤细胞A375 MIA/CD-RAP基因启动子的克隆及其鉴定

目的:克隆MIA/CD-RAP的启动子序列并对其进行酶切和DNA测序鉴定。方法:从培养的人黑素瘤细胞A375基因组DNA中扩增出MIA/CD-RAP启动子序列,并将其克隆到载体pGL2-Ba-sic上,再经酶切及PCR扩增鉴定和测序。结果:酶切电泳和DNA测序结果表明,已成功克隆了人黑素瘤MIA/CD-RAP启动子序列。结论:人黑素瘤MIA/CD-RAP启动子序列的克隆,可为进一步研究MIA/CD-RAP基因的表达调控奠定基础。

黑素瘤MIA启动子介导的CD/TK双自杀基因重组腺病毒系统选择性杀伤黑素瘤细胞

目的:探讨黑素瘤抑制蛋白(melanoma inhibitory activity,MIA)启动子介导的CD/TK双自杀基因重组腺病毒系统,对黑素瘤细胞A375的选择性杀伤作用。方法:构建腺病毒穿梭质粒pAdrMIA-CD/TK,在293细胞内包装、扩增、纯化后,体外转染人表皮黑素细胞HeMa-LP、黑素瘤细胞A375和人宫颈癌细胞HeLa,RT-PCR检测基因CD和TK片段,加入前体药物5-氟胞嘧啶(5-FC)和/或丙氧鸟苷(GCV),用MTT法测定该体系对细胞株的杀伤效应。结果:制备的病毒滴度为1×1011pfu/L,用100m.o.i.重组腺病毒感染细胞后,发现目的基因CD和TK片段可在黑素瘤细胞中表达,转染目的基因的黑素瘤细胞A375对5-FC和GCV具有一定的敏感性,细胞存活率较对照组显著下降(P<0.05),联合应用两种药物对A375细胞的杀伤作用较单用一种显著增强(P<0.05)。对人表皮黑素细胞HEMa-LP和人宫颈癌细胞HeLa无显著杀伤作用。结论:构建的黑素瘤MIA启动子介导的CD/TK双自杀基因重组腺病毒系统,在体外可选择性杀伤黑素瘤细胞A375。

疏肝祛瘀针刺法治疗黄褐斑45例疗效观察

黄褐斑为临床上常见而又难以治愈的皮肤病之一。多发于青年女性。目前西医治疗主要包括抑制黑素细胞活性、抑制黑素合成、去除已经生成的黑素、破坏黑素颗粒以及运用具有防光效应的遮光剂等，但常因服用药物麻烦、疗效不确切及药物副作用等缺点，使病人难以接受。近年来笔者采用疏肝祛瘀针刺法治疗黄褐斑取得了较好疗效，现报道如下。

Inhibitory effects of N-(4-hydrophenyl) retinamide on liver cancer and malignant melanoma cells

AIM: To investigate the effect of N-(4-hydrophenyl) retinamide (4-HPR), the derivative of retinoic acid, on inhibition of migration, invasion, cell growth, and induction of apoptosis in hepatocellular carcinoma cells (HCCs) and malignant melanoma cells.METHODS: 4-HPR was chemically synthesized. Cellular migration and invasion were assayed by Borden chamber experiment. Cell growth was assayed by MTT chromometry.Apoptosis effect was measured using Hoechst 32258 staining and flow cytometry. Gene transfection was performed with lipofectamine.RESULTS: We observed that the migration of HCC and melanoma cells was significantly suppressed by 4-HPR and the migration cells were reduced to 58±5.03 (control 201±27.2, P＜0.05, n = 4) in SMMC 7721-k3 HCC, and to 254±25.04 (control 302±30.1, P＜0.05, n = 4) in melanoma cells after 6-h incubation with 4-HPR. The invasion through reconstituted basement membrane was also significantly reduced by 4-HPR treatment to 11.2±3.3 in SMMC 7721-k3 HCC (control 27±13.1), and to 24.3±3.2 in melanoma cells (control 67.5±10.1, P＜0.05, n = 3). Cell growth, especially in melanoma cells, was also significantly inhibited.Furthermore, 3 μmol/L of 4-HPR induced apoptosis in B16 melanoma cells (37.11±0.94%) more significantly than all-trans retinoic acid (P＜0.05), but it failed to induce apoptosis in SMMC 7721-k3 HCC. The mechanism for 4-HPR-induced apoptosis was not clear, but we observed that 4-HPR could regulate p27kip1, and overexpression of cerebroside sulfotransferase (CST) diminished the apoptosis induced by 4-HPR in melanoma cells.CONCLUSION: 4-HPR is a potent inhibitor of HCC migration and inducer of melanoma cell apoptosis. CST and p27kip1 expression might be associated with 4-HPR-induced apoptosis.

Melanoma differentiation-associated gene-7, MDA-7/IL-24, selectively induces growth suppression, apoptosis in human hepatocellular carcinoma cell line HepG2 by replication-incompetent adenovirus vector

AIM： To investigate the effect of replication-incompetent adenovirus vector expressing MDA-7/IL-24 on tumor growth and apoptosis in human hepatocellular carcinoma （HCC） cell line HepG2 and normal liver cell line L02. METHODS： We constructed the recombinant replication-incornpetent Ad.rnda-7 virus vector and infected it into the human HCC cell line HepG2 and normal liver cell line L02. RT-PCR was performed to detect the rnRNA expressing in cells, by ELISA was used to detect MDA-7/IL-24 protein expression in the culture supernatant. The effect of apoptosis induced by Ad.rnda-7 was confirmed by Hoechst staining and flow cytometry assay with Annexin-V and PI staining. MTT assay was used to determine growth inhibition of HepG2 cells, and cell-cycle and hypodiploidy analyses were performed by flow cytometry. RESULTS： Recombinant replication-defective virus expressing MDA-7/IL-24 was constructed successfully. RTPCR showed that the Ad.rnda-7 could mediate the expression of the exogenous gene MDA-7/IL-24 into HepG2 and L02. The concentration of MDA-7/IL-24 protein in supernatant was 130 pg/mL and 110 pg/mL in Ad.rnda-7-infected L02 and HepG2 ceils, respectively. Ad.mda-7 infection obviously induced apoptosis （from 2.604±0.72% to 33.64±13.2%, P=0.00012） and growth suppression in HepG2 （inhibition ratio IR=68%） and an increase in the percentage of specific cancer cell types at the G2/M phase of the cell cycle （from 6.44% to 32.29%, P〈 0.01）, but not in L02 cells.CONCLUSION： These results confirm selectively induction of apoptosis and growth suppression by the mda-7/ IL-24 gene with replication-incompetent adenovirus vector in human hepatocellular carcinoma cell line HepG2.

Evaluation of graphite depressants in a poly-metallic sulfide flotation circuit

Synthetic dyes are commonly used for graphite depression in poly-metallic flotation circuits; however,these dyes can be very expensive. The aim of this study is to evaluate performance of certain low-cost alternative depressants for a complex lead-zinc(Pb-Zn) ore rich in graphite(Gr-C) on a conventional mini pilot-scale flotation circuit. The reagents used were commercial and industrial grade starch; agro-based waste-sugarcane bagasse and charred(burnt) bagasse powder. The primary evaluation criteria were quality(grades) of lead and zinc concentrates, their recoveries(%), and graphite rejection(%) in the tails.Benchmark tests using nigrosine as graphite depressant showed 94.3% rejection of Gr-C. The results with commercial starch were found as effective with 93.8% graphite rejection. Furthermore, bagasse powder showed potential in improving product quality(36.4% and 65.6% Pb grade and recovery) with an intermediate effectiveness in graphite rejection(85.6%). The order of effectiveness in Gr-C rejection follows nigrosine % commercial starch > bagasse > industrial starch > charred bagasse. In addition, the effect these depressants on silver(byproduct) grade and recovery was also investigated.

Migration-associated secretion of melanoma inhibitory activity at the cell rear is supported by KCa3.1 potassium channels

Malignant melanoma, characterized by invasive local growth and early formation of metastases, is the most aggressive type of skin cancer. Melanoma inhibitory activity （MIA）, secreted by malignant melanoma cells, interacts with the cell adhesion receptors, integrins a4131 and 05131, facilitating cell detachment and promoting formation of me- tastases. In the present study, we demonstrate that MIA secretion is confined to the rear end of migrating cells, while in non-migrating cells MIA accumulates in the actin cortex. MIA protein takes a conventional secretory pathway including coat protein complex I （COPI）- and coat protein complex II （COPII）-dependent protein transport to the cell periphery, where its final release depends on intracellular Ca2＋ ions. Interestingly, the Ca2＋-activated K＋-channel, subfamily N, member 4 （KCa3.1）, known to be active at the rear end of migrating cells, was found to support MIA secretion. Secretion was diminished by the specific KCa3.1 channel inhibitor TRAM-34 and by expression of dominant- negative mutants of the channel. In summary, we have elucidated the migration-associated transport of MIA protein to the cell rear and also disclosed a new mechanism by which KCa3.1 potassium channels promote cell migration.

北向单侧采光办公房间的非视觉效应研究

近年来,学者们逐渐开始关注建筑采光环境中非视觉效应所引起的健康风险。然而,目前采光设计时仅考虑视觉作业,对既有建筑采光环境的非视觉效应缺乏了解。为解决该问题,本文选择满足GB 50033-2013《建筑采光设计标准》Ⅲ设计等级的北向单侧天然采光办公房间进行实测,利用平视采光外窗的人眼处垂直照度与褪黑素抑制百分比的剂量效应关系评价该办公房间的非视觉效应,结果表明在阴天条件下,该房间褪黑素抑制水平处于临界状态,即距窗较远的进深区域部分时段褪黑素抑制百分比难以达到100%。此外,还对采光设计标准中单侧采光房间Ⅰ~Ⅴ设计等级的非视觉效应进行了评价,各等级的褪黑素抑制百分比分别为100%、100%、71%~100%、38%~60%、5%~16%,这表明Ⅰ与Ⅱ设计等级可充分满足的褪黑素抑制需求、Ⅲ设计等级则处于临界状态、而Ⅳ与Ⅴ设计等级的褪黑素抑制水平较低。

常见美容方法的是与非

爱美之心人皆有之,我们也经常可以从各种美容机构的广告里听到各种排毒、美白、美容养颜,由此也有不少女性在美容院中一掷千金只为换回年轻的肌肤,可是排毒、美白、美容养颜的产品和技术是否真的可靠呢？快速美白亚洲人皮肤天生偏黄,对美白有着执着的追求,美白产品更是数不胜数。据悉,目前真正国内及FDA认证的具有抑制黑素生长及分泌的有：氢醌、氨甲环酸、维生素C。