肝细胞癌患者树突状细胞免疫功能的研究

 目的：研究肝细胞癌患者树突状细胞(DCs)的免疫功能变化。方法：从人外周血中分离单个核细胞（PBMC），用GM-CSF和IL-4体外诱导、培养和扩增DCs，流式细胞仪测定DCs表面分子的表达，分别用ELISA、ELISPOT、MTT法测定HCC患者和健康人的DCs分泌IL-12；T淋巴细胞分泌IFN-γ和T淋巴细胞的增殖反应。结果：GM-CSF和IL-4能成功地体外诱导、培养和扩增DCs。HCC患者DCs的CD86表达明显低于健康对照者（91.7％ vs83.5%, P<0.05）。HCC患者DCs分泌IL-12与健康者差异不显著［（324.6±171.0）ng/L vs（436.5±142.7）ng/L, P>0.05］，接受内毒素LPS刺激后，差异更明显［（478.6±142.7）ng/L vs（630.0±151.9）ng/L, P<0.05］。DCs刺激T淋巴细胞分泌IFN-γ 的量在HCC患者明显低于健康者［（133.4±51.2）103U/L vs (183.0± 60.2)103U/L, P<0.05］，DCs刺激T淋巴细胞的增殖能力在HCC患者明显低于健康者（2.3±0.7 vs3.5±0.8, P<0.01）。CTL杀伤实验表明，当效靶比为 10∶〖KG-*2〗1 和 20∶〖KG-*2〗1 时，HCC组的CTL对肝癌细胞Hep G2的杀伤活性明显低于健康组。结论： HCC患者DCs功能和T淋巴细胞功能低于健康者，HCC患者DCs诱导的CTL杀伤活性亦明显低于健康者。     

IL-10抑制AVP诱导大鼠心脏成纤维细胞CaN活性

目的：研究白细胞介素10（IL-10）对精氨酸血管加压素(AVP)诱导大鼠心脏成纤维细胞(CFs)钙调神经磷酸酶（CaN）活性的影响。方法:用培养的新生SD大鼠CFs，四氮唑盐(MTT)比色法检测CFs增殖，流式细胞仪检测细胞周期，分光光度法测定CaN活性。结果:（1）10-7 mol/L AVP组CFs的吸光度（A490）明显高于对照组（P<0.01）。IL-10呈浓度依赖性下调 AVP诱导的CFs的A490值（P<0.05或P<0.01）。IL-10本身对CFs的增殖无抑制作用。（2）10-7 mol/L AVP组CFs的S期百分率及增殖指数均明显高于对照组（均P<0.01）；10-6 g/L IL-10+10-7 mol/LAVP组S期百分率及增殖指数均明显低于AVP组（P<0.01）。（3）10-7 mol/L AVP组CaN活性明显高于对照组（P<0.01）；10-8、10-7、10-6和10-5g/L +10-7 mol/L AVP组的CaN活性均明显低于AVP组（P<0.01），但仍高于对照组（P<0.01或P<0.05）。IL-10呈浓度依赖性下调 AVP诱导CFs的CaN活性。结论:IL-10具有抑制AVP诱导CFs增殖和下调CFs的CaN活性的作用，这可能对预防和逆转心脏重构有一定的价值。     

正丁酸钠对人肝癌细胞的生物学效应

目的:探讨正丁酸钠(SB)对人肝癌细胞的生物学效应.方法:以1mmol/L的SB作用于培养的人肝癌细胞,观察肝癌细胞某些生物学行为的变化.结果:在SB作用下,人肝癌细胞生长抑制.细胞形态改变,ConA凝集现象消失,琼脂集落形成率下降(p<0.001),染色体主流范围变窄,二倍体及亚二倍体比例升高,~3H—TdR掺人率、细胞DNA含量下降与对照比较有显著差异(p<0.01).结论:人肝癌细胞在丁酸钠作用下,某些恶性生物学行为可发生部分逆转.     

食管癌相关基因4在肝细胞肝癌中的表达和功能

目的了解人食管癌相关基因4（ ECRG4）在人肝细胞肝癌组织和肝癌细胞系中的表达情况,探讨ECRG4对肝癌细胞增殖、凋亡以及迁移能力的影响。方法提取24例配对肝细胞肝癌组织和癌旁肝组织的总RNA以及正常肝细胞系QSG7701和肝癌细胞系HepG2的总RNA和总蛋白,分别利用即时荧光定量PCR（ RT-qPCR）和Western blot法检测ECRG4的表达;构建ECRG4的真核表达质粒ECRG4-pcDNA3.1,转染肝癌细胞系HepG2,进行体外的细胞增殖、凋亡以及迁移实验。结果与癌旁肝组织相比,在肝细胞肝癌组织中ECRG4 mRNA表达下调或缺失（98.5％,23／24,P＜0.01）;和正常肝细胞系QSG7701相比,ECRG4 mRNA在肝癌细胞系HepG2中的表达明显下调（ P＜0.05）并且其蛋白表达水平也明显降低。转染ECRG4表达质粒后HepG2细胞的增殖和迁移能力明显低于对照组（P＜0.05）,细胞凋亡率明显高于对照组（P＜0.05）。结论 ECRG4在肝细胞肝癌中表达下调,在体外肝癌细胞系中过表达ECRG4可抑制细胞的增殖和迁移,促进细胞凋亡,ECRG4可能是肝细胞肝癌潜在的抑癌基因,有望为肝细胞肝癌分子靶向治疗提供新策略。     

miRNA-122促进小鼠胚胎干细胞源性肝前体细胞的分化与成熟

 目的： 通过miRNA-122 (miR-122)转染胚胎干细胞（ESCs）源性肝前体细胞,观察及评估其是否能推进ESCs向肝细胞的分化过程。方法：采用丁酸钠、成纤维细胞生长因子4(FGF-4)及地塞米松(Dex)联合序贯诱导小鼠ESCs使之初步分化为肝前体细胞,然后利用Gateway技术构建出pAV.Ex1d-CMV>miR122/IRES/eGFP重组腺病毒表达载体,使其转染贴壁诱导第9天的小鼠ESCs源性肝前体细胞,获得稳定高表达miR-122的细胞后继续培养。采用荧光显微镜观察转染后细胞形态的变化;使用real-time RT-PCR检测肝特异性基因的表达;免疫荧光检测肝特异性蛋白表达水平;吲哚氰绿摄取实验、糖原染色及尿素合成功能实验检测肝细胞功能。结果：丁酸钠、FGF-4及Dex联合序贯诱导可将小鼠ESCs成功地诱导为肝前体细胞。转染miR-122后,细胞在形态学上更接近成熟肝细胞;肝特异性基因白蛋白(ALB)、甲状腺素运载蛋白、α1抗胰蛋白酶、葡萄糖-6-磷酸酶、细胞角蛋白8、胆固醇7α-羟化酶及细胞色素P450 3A4的mRNA表达均增高;肝特异性蛋白ALB及细胞角蛋白18均表达增高,而甲胎蛋白表达降低;吲哚氰绿摄取实验、糖原染色及尿素合成功能实验结果均显示转染miR-122后肝细胞功能较对照组明显增强。结论：丁酸钠、FGF-4及Dex联合序贯诱导可将小鼠ESCs成功地诱导为肝前体细胞;miR-122能够有效地促进小鼠肝前体细胞的分化与成熟。     

促酰化蛋白诱导3T3-F442A前脂肪细胞分化的研究

目的：探讨新型脂源性激素促酰化蛋白（ASP）是否具有诱导前脂肪细胞分化的作用。方法： 以3T3-F442A前脂肪细胞为研究对象，通过形态学观察、油红染色测定脂肪细胞分化率，测定脂肪细胞甘油三酯合成率和甘油三酯总量，采用［3H］-胸腺嘧啶掺入法，反映ASP诱导3T3-F442A前脂肪细胞分化过程中克隆增殖的情况，并与经典的诱导分化剂胰岛素比较。结果： (1)单独的ASP刺激，即可诱导3T3-F442A 前脂肪细胞向成熟脂肪细胞的形态转变，且分化率较高，与胰岛素刺激3T3-F442A 前脂肪细胞分化相比，无显著差异（P>0.05）。(2)ASP促进3T3-F442A前脂肪细胞的甘油三酯合成，并增加细胞甘油三酯的总量，均明显高于对照组(P<0.05)，而与胰岛素组相比无显著差异（P>0.05）。(3)分化诱导24 h，ASP组［3H］-胸腺嘧啶掺入率是对照组的223%（P<0.01），胰岛素组［3H］-胸腺嘧啶掺入是对照组的589%（P<0.01）。结论： 新型的脂源性激素ASP具有诱导前脂肪细胞分化为成熟脂肪细胞的生物学作用。     

肝匀浆上清液诱导人脐带间充质干细胞分化为肝样细胞

背景：前期实验证明大鼠肝组织匀浆上清液能诱导人脐带间充质干细胞发生形态学变化,但诱导后细胞是否具有肝细胞的表型和功能特征尚不清楚。 目的：以人脐带间充质干细胞为研究对象,对肝组织匀浆上清液定向诱导分化后细胞是否具有肝细胞的表型和功能特性进行初步验证。 方法：选取第3代人脐带间充质干细胞,采用肝组织匀浆上清液作为诱导培养基,进行如下分组：单纯肝组织匀浆上清液作为阴性对照组,QSG-7701人肝细胞株作为阳性对照组,基础培养的人脐带间充质干细胞作为标准对照组;肝组织匀浆上清液处理3,5,7 d诱导组;在蛋白水平和转录水平对肝细胞特异性标志物甲胎蛋白、角蛋白18和色氨酸2,3-加双氧酶进行检测。 结果与结论：经肝组织匀浆上清液诱导后,细胞形态发生了显著变化,随诱导时间延长梭形干细胞逐渐减少,细胞呈三角形,多角形或不规则形。肝组织匀浆上清液诱导干细胞3,5,7 d后肝细胞标志物甲胎蛋白、人细胞角蛋白18和色氨酸2,3-加双氧酶在蛋白和mRNA水平均显著高于标准对照组(P < 0.01)。实验结果提示在体外经肝组织匀浆上清液诱导后,人脐带间充质干细胞能够分化为具有肝细胞特异性标记的肝样细胞,并具有部分肝细胞功能。     

人肝癌与癌旁肝AFP阳性细胞形态的图象分析及增殖活性的研究

采用免疫组化、双标免疫组化及图象分析技术，研究了２７例人体肝细胞癌与癌旁肝甲胎蛋白（ＡＦＰ）阳性细胞的量化形态特征及以其增殖细胞核抗原（ＰＣＮＡ）标记率显示的增殖活性。主要结果显示：（１）ＡＦＰ阳性与阴性癌及癌旁近癌肝细胞间难以用量化形态标准区分（Ｐ＞０．０５）；核面积与核浆比提示癌旁ＡＦＰ阳性肝细胞可能是尚无表型转化的癌前细胞。（２）ＡＦＰ阳性癌细胞ＰＣＮＡ的平均标记率（１５．７０％±６．２５％）显著低于肝癌的平均标记率（Ｐ＜０．０１），表明大多数ＡＦＰ阳性癌细胞处于非增殖状态。（３）癌旁近癌肝可见ＡＦＰ与ＰＣＮＡ双标阳性肝细胞。     

eIF4H低表达对MEL细胞增殖和红系分化的影响简

 目的 探究eIF4H低表达对MEL细胞增殖和红系分化的影响。方法 重组质粒载体eIF4H-shRNA-1或eIF4H-shRNA-2与pCMV-VSVG和pCMV-dR8.2共同转染HEK293T细胞,获得可使eIF4H低表达的慢病毒。慢病毒侵染MEL细胞,获得eIF4H低表达的MEL细胞稳定株,Western blot检测干扰效果。MTT检测细胞增殖;流式细胞术检测细胞周期;联苯胺染色观察红系分化情况。结果 与对照组相比较,eIF4H-shRNA-2组细胞eIF4H表达明显下调,细胞增殖能力明显减弱（P<0.01）,G1期细胞比例明显增加;经丁酸钠处理后eIF4H-shRNA-2组联苯胺阳性细胞明显增加（P<0.01）。结论 成功构建eIF4H低表达MEL稳定株;eIF4H低表达抑制MEL细胞增殖能力;eIF4H低表达不能诱导MEL细胞红系分化,但可促进丁酸钠诱导MEL细胞红系分化。     

沉默HIF-1α基因表达对大鼠肝癌细胞增殖的影响

 目的： 探讨沉默缺氧诱导因子　1α（HIF-1α）基因表达对缺氧状态下肝癌细胞增殖的影响。方法：选用大鼠CBRH-7919肝癌细胞株作为研究对象,利用氯化钴（CoCl2）建立缺氧模型。制备3组特异性较强的HIF-1α siRNA-脂质体复合物,转染于肝癌细胞。利用real-time RT-PCR、Western blotting等方法分别检测肝癌细胞HIF-1α、血管内皮生长因子(VEGF)、p21和cyclin D1在mRNA和（或）蛋白水平的表达。MTT及BrdU 掺入实验检测细胞增殖的变化。结果：缺氧条件下,肝癌细胞的HIF-1α和VEGF mRNA及蛋白表达显著增多（P<0.05）。HIF-1α基因表达沉默后,HIF-1α、VEGF及cyclin D1 mRNA和（或）蛋白表达明显减少（P<0.05）,p21蛋白表达明显增加（P<0.05）。HIF-1α siRNA转染组的BrdU阳性细胞比例明显少于对照组(P<0.05)。结论：沉默HIF-1α基因表达对缺氧状态下肝癌细胞的增殖有显著抑制作用。     

Schizophrenia in a North Swedish geographical isolate, 1900–1977. Epidemiology, genetics and biochemistry

Over 200 schizophrenic patients belonging to three major and interrelated pedigree complexes have been investigated over the past 30 years in a North Swedish geographically isolated population, presently numbering about 6,000. An intensive investigation of a number of biochemical correlates and genetic markers in a few selected families belonging to one of the major pedigrees has indicated new strategies for the current research program.
Schizophrenia, as defined operationally, is significantly associated with decreased activities of two enzymes (1) blood platelet monoamine oxidase, (2) plasma dopamine-β-hydroxylase, and (3) with the genetic marker Gc² (group specific antigen). Both enzymes are subject to genetic variation. A positive score for linkage between schizophrenia and low plasma DBH activity has been calculated, but, so far, available data are insufficient for discrimination between linkage and partial contribution of genetically controlled low plasma DBH to the pathogenesis of the disease. Alternatively, both mechanisms could be involved.
As a model for continued research, schizophrenia is explained as based on a double dominant-recessive genotype (Aabb), representing a vulnerability which in about 50 % of cases develops into clinical schizophrenia. It is suggested that the dominant mutation (A) operates on or affects MAO activity, and that the recessive genotype (bb) is instrumental in low variates of DBH activity and very likely such variates within the normal range of physiological variation. Moreover, it is suggested that the combined effects of MAO- and DBH-reduced efficiency on the metabolism of e.g. dopamine could be an essential pathogenic mechanism for the schizophrenic illness which is segregating in this population.     

Renal dysplasia and asplenia in two sibs

A family is reported in which two sibs, one male and the other female, both died within 24 hours of birth with enlarged polycystic kidneys. Postmortem histology in the second child showed gross renal dysplasia. In both children the pancreas was enlarged, nodular and cystic but the liver appeared macroscopically normal. In the second child, histological examination confirmed pancreatic fibrosis with cystic dilation of ducts, but showed portal fibrosis with bile duct proliferation in the liver.
This combination of findings is very reminiscent of those in a girl and her brother reported by Ivemark et al. (1959). The children reported here also showed absence or hypoplasia of the spleen, cardiac anomalies and other features of the Ivemark syndrome (Ivemark 1955), a quite different, usually sporadic, congenital disorder. It is suggested that the children described here have a distinct lethal congenital disorder, probably inherited in an autosomal recessive manner.     

HLA in North Colombia Chimila Amerindians

HLA-A,-B,-C,-DRB1 and -DQB1 alleles have been studied in Chimila Amerindians from Sabana de San Angel (North Colombian Coast) by using high resolution molecular typing. A frequent extended haplotype was found:HLA-A*24:02-B*51:10-C*15:02-BRB1*04:07-DQB1*03:02 (28.7%) which has also been described in Amerinndian Mayos Mexican population (Mexico, California Gulf, Pacific Ocean). Other haplotypes had already been found in Amerindians from Mexico (Pacific and Atlantic Coast), Peru (highlands and Amazon Basin), Bolivia and North USA. A geographic pattern according to HLA allele or haplotype frequencies is lacking in Amerindians, as already known. Also, five new extended haplotypes were found in Chimila Amerindians. Their HLA-A*24:02 high frequencies characteristic is shared with aboriginal populations of Taiwan; also, HLA-C*01:02 high frequencies are found in New Zealand Maoris, New Caledonians and Kimberly Aborigines from Australia. Finally, this study may show a model of evolutionary factors acting and rising one HLA allele frequency (-A*24:02), but not in others that belong to the same or different HLA loci.     

The universal muscular chamber. A basic unit of the genitourinary, cardiovascular, gastro-intestinal, skeletal, cutaneous, respiratory and other systems, endocameral hypertension; and spasm, the key to their idiopathic diseases and arthropathies

Starting with the integument, we see many organs are contractile sacs or multiples thereof, which tubes or bags constitute the major part of the entire body. Recognition of this basic unit and its characteristics sheds new light, individually and collectively, on many disorders previously considered unrelated. Muscular tears and perforations develop in the walls of these chambers, being no way peculiar to those organs, wherein, hydrochloric acid occurs. So, it is not necessary to explain the absence of excessive acid from patients who exhibit holes in the gastric, uterine, aortic, duodenal, rectal, pulmonary, retina, and other walls. Muscle, not acid is the great common factor relating idiopathic disorders in the gastrointestinal tract to each other and to similar diseases in other systems. When the units are linked together, the lesions tend to appear as arthropathies, i.e. at the joints. Rephrasing common-place observations, frees us from conventional, conceptual cul-de-sacs. An observation is only as good as its interpretation, so all possibilities must be considered, otherwise, we will remain blinded by our misconceptions.     

Der Einfluß von Nahrungsmitteln und Rauchen auf die klinisch-chemische Diagnostik von Phäochromozytom,Neuroblastom und Karzinoid-Syndrom

Zusammenfassung Der Einfluß von verschiedenen Nahrungsmitteln auf Methoden zur Bestimmung von Adrenalin (AD), Noradrenalin (NA), Vanillinmandelsäure (VMS), Metanephrinen (MN), Homovanillinsäure (HVS) und 5-Hydroxyindolessigsäure (5-HIE) im 24 h-Harn zur Diagnose des Phäochromozytoms bzw. Karzinoid-Syndroms wurde untersucht. Die in die Untersuchung einbezogenen Nahrungsmittel waren: Tee, Kaffee, Mandeln, Ananas, Käse, Walnüsse, Vanillepudding, Bananen, Tomaten und Milchschokolade. Außerdem wurde der Einfluß des Zigarettenrauchens auf die Bestimmung von AD, NA, VMS und MN untersucht.Walnüsse führten zu einer starken Erhöhung der 5-HIE-Ausscheidung. Bananen erhöhten die Ausscheidung von AD, NA, VMS, MN und 5-HIE. Kaffee und Ananas bewirkten eine geringe Zunahme der MN-Werte. Rauchen von 20–30 Zigaretten/Tag beeinflußte keine der vier Variablen.Wenn die beschriebenen Methoden benutzt werden, sollte lediglich auf den Verzehr von Bananen und Walnüssen vor und während der Harnsammelperioden verzichtet werden, da die oberen Normgrenzen im Harn überschritten werden könnten. Ein Verzicht auf Kaffee und Ananas in normalen Mengen ist nicht erforderlich. Es besteht kein Anlaß, weiterhin die bisherigen umfangreichen Restriktionen der übrigen Nahrungsmittel beizubehalten.     

A comparative study of the effects of dimebon,obsidan, finoptin,and cordaron on the functional state of ischemic focus and size of necrotic zone in experimental myocardial infarction

Dimebon, an antihistamine agent, exerts a moderate antianginal effect, improving the function of ischemic focus in the myocardium and decreasing the necrotic zone in experimental myocardial infarction. Dimebon is less active than obsidan, finoptin (except for the size of the necrotic zone), and cordaron. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 122, No. 12, pp. 642–644, December, 1996     

Effects of sex steroid hormones on lipid peroxidation and glutathione antioxidant system in rat skin

Effects of estradiol and testosterone on the intensity of lipid peroxidation and contents of glutathione redox system components in the dermis and epidermis of rat skin were studied. Only estradiol induced considerable dose-dependent and tissue-specific biphasic antioxidant effects on the skin. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 128, No. 12, pp. 663–666, December, 1999     

Adhesion and growth of CaCo2 cells on surface-modified PEEK substrata

A series of surface-functionalized poly(ether ether ketone) (PEEK) films has been prepared by selective wet-chemistry; they are hydroxylated polymer (PEEK-OH) obtained by reduction, aminated polymer (PEEK-[]-NH2) prepared by coupling a diisocyanate reagent to PEEKOH (PEEK-[]-NCO) followed by hydrolysis, and carboxylated and aminocarboxylated polymers (PEEK-[]-GABA and PEEK-Lysine) resulting from the coupling of aminoacids to PEEK-[]-NCO. The aminated and carboxylated substrata promoted the adhesion and growth of CaCo2 cells in the presence of serum. Fibronectin (FN), an extra-cellular matrix protein, has been covalently fixed and/or adsorbed on various PEEK substrata, in the presence or not of a polymeric surfactant (Pluronic F68). The performances of the FN-grafted substrata (PEEK-[]-FN(1) and PEEK-[]-FN(2)) were significantly higher than those of reference substrata simply coated with FN (PEEK-OH(+FN)(1) and (2), PEEK-[]-NH2(+FN)(1) and (2)), considering the adhesion and spreading of CaCo2 cells in the absence of serum. Moreover, the stability of the adherent cells on the FN-adsorbed substrata dramatically depended on the experimental conditions applied during the PEEK coating with FN.