miR-98在涎腺腺样囊性癌组织中的表达及其对ACC-M细胞增殖、迁移的影响

目的探讨miR-98在涎腺腺样囊性癌组织中的表达及其对腺样囊性癌细胞系ACC-M增殖以及迁移能力的影响。方法通过实时PCR检测5例涎腺腺样囊性涎组织及癌旁正常组织miR-98的表达水平;将miR-98模拟物瞬时转入高转移腺样囊性癌细胞系(ACC-M)使miR-98过表达,并通过实时PCR方法验证;流式细胞仪分析转染后ACC-M细胞周期的改变,甲基噻唑基四唑比色法检测转染后ACC-M细胞的增殖,划痕实验检测转染后ACC-M细胞的迁移能力,免疫印迹法检测转染后ACC-M细胞细胞周期蛋白CyclinB与CDC2的表达变化。结果 miR-98在涎腺腺样囊性癌组织中的表达水平显著低于癌旁正常组织,P0.01。转染miR-98模拟物能够显著上调ACC-M细胞miR-98的表达水平,ACC-M细胞的S期比例明显下降,而G 0/G 1期的细胞明显增多,增殖受到明显抑制,迁移能力下降,显著降低ACC-M细胞CyclinB与CDC2的蛋白表达水平。结论过表达miR-98能够抑制涎腺腺样囊性癌细胞的增殖与迁移能力。     

趋化因子SDF-1α及其受体介导涎腺腺样囊性癌细胞的趋化与侵袭

探讨趋化因子SDF-1α及其受体CXCR4对涎腺腺样囊性癌细胞趋化与侵袭活性的促进作用。采用RT-PCR法检测涎腺腺样囊性癌肺高、低转移细胞株ACC-M和ACC-2中CXCR4 mRNA的表达;Boyden趋化小室法检测在SDF-1α作用下ACC-M细胞和ACC-2细胞的趋化与侵袭活性;检测CXCR4 mAb对ACC-M细胞和ACC-2细胞趋化活性和侵袭活性的抑制作用。结果显示:2株涎腺腺样囊性癌细胞中均存在不同程度CXCR4 mRNA的表达,其在肺高转移细胞株ACC-M中的表达显著高于肺低转移细胞株ACC-2;SDF-1α对2种细胞均具有趋化活性和侵袭活性,且对ACC-M细胞的趋化活性和侵袭活性显著强于ACC-2细胞;CXCR4 mAb能够抑制涎腺腺样囊性癌细胞的这种趋化活性和侵袭活性。趋化因子SDF-1α及其受体CXCR4能够介导涎腺腺样囊性癌细胞的趋化与侵袭。     

miRNA-542-3p对胃腺癌细胞侵袭转移能力的影响

目的研究microRNA-542-3p(miR-542-3p)在人胃腺癌细胞系及组织中的表达水平及对胃腺癌细胞侵袭转移的影响。方法通过RT-PCR检测miR-542-3p在人正常胃粘膜上皮细胞GES-1及人胃癌细胞系SGC-7901和BGC-823中的表达水平;同时应用RT-PCR检测miR-542-3p在50组患者胃腺癌和癌旁组织中的表达差异。体外实验通过外源转染miR-542-3p的模拟物和抑制剂,分别于24,36,48h检测SGC-7901的转染效率,选取最佳转染时间。应用划痕实验和Transwell评估细胞侵袭转移能力的变化。结果胃腺癌细胞系SGC-7901和BGC-823中miR-542-3p的表达水平低于其在GSE-1中的表达水平(P0.05);miR-542-3p在癌组织中表达低于癌旁组织(P0.05)。24 h后,miR-542-3p模拟物组侵袭转移能力显著低于对照组(P0.05),而miR-542-3p抑制剂组侵袭转移能力显著高于对照组(P0.05)。结论miR-542-3p在多种人胃癌细胞和胃腺癌组织中呈低表达,miR-542-3p抑制胃腺癌SGC7901细胞的侵袭转移能力。     

卵巢癌组织中miR-574-5p的表达及其下调后对SKOV-3细胞增殖与侵袭的影响

目的观察miR-574-5p在卵巢组织中的表达及下调miR-574-5p表达对卵巢癌细胞株SKOV-3增殖与侵袭的影响。方法实时PCR检测41例卵巢癌组织及对应癌旁正常卵巢组织miR-574-5p的表达;随机将SKOV-3细胞分为空白对照组、阴性对照组和miR-574-5p抑制物组,分别用MTT方法与Transwell细胞侵袭实验检测各组SKOV3细胞的增殖与侵袭能力变化。结果卵巢癌组织中miR-574-5p相对表达量为1.29±0.18,显著高于癌旁正常卵巢组织的0.51±0.07(P0.01);转染miR-574-5p抑制物后,SKOV3细胞的增殖能力明显下降,穿膜能力亦明显减弱(P0.01)。结论卵巢癌组织中miR-574-5p的表达水平异常增高,抑制miR-574-5p的表达能够降低SKOV3细胞的增殖与侵袭。     

miR-204-5p抑制乳腺癌细胞增殖、迁移、侵袭及对上皮-间质转化的影响

目的 探索miRNA-204-5p在乳腺癌组织和细胞中的表达,探讨miR-204-5p对乳腺癌细胞增殖、迁移、侵袭和上皮-间质转化(EMT)的影响.方法 qRT-PCR检测乳腺癌患者癌和癌旁组织,乳腺癌细胞和正常乳腺上皮细胞中miR-204-5p的表达;将miR-204-5p模拟物和阴性对照分别转染MCF-7细胞,并检测转染效率;CCK-8法检测转染对细胞增殖的影响;Transwell迁移实验和侵袭实验分别检测转染miR-204-5p后乳腺癌细胞迁移和侵袭能力的变化;Western blot检测上皮-间质转化(EMT)相关蛋白的表达.结果 miR-204-5p在乳腺癌组织表达水平显著下调,miR-204-5p在多种乳腺癌细胞株如MCF-7细胞表达量下调.miR-204-5p模拟物转染抑制MCF-7细胞增殖、迁移、细胞侵袭,E-cadhesin表达水平升高,Vimentin降低.结论 过表达MiR-204-5p对乳腺癌有潜在的治疗作用.     

miR-133a-3p 通过抑制氧化还原因子1 抑制肝癌细胞恶性生物学行为

目的:通过多种实验途径探讨miR-133a-3p通过调节氧化还原因子1(Ref-1)对肝癌细胞恶性生物学行为的影响。方法:收集2017年2月～2018年12月来本院就诊的肝癌手术患者的癌组织和癌旁相邻正常组织标本,用qRT-PCR检测miR-133a-3p在癌组织和癌旁相邻正常组织中以及在肝癌细胞系HepG2、Huh-7、SK-Hep1、SMMC-7721和肝正常细胞Hep-3B中的表达;构建miR-133a-3p-mimics(miR-mimics组、miR-NC转染质粒组和正常对照组(Control),qRT-PCR检测转染前后miR-133a-3p在HepG2中的表达;CCK-8和克隆形成实验检测HepG2细胞转染前后增殖能力变化;细胞划痕和Transwell实验分别检测转染前后HepG2细胞的迁移和侵袭能力变化;RNA转录组测序检测HepG2细胞转染前后转录组RNA表达差异并作通路富集分析;qRT-PCR检测筛选RNA-seq中RNA表达差异较大的基因;qRT-PCR和Western blot检测Ref-1在HepG2细胞转染前后中的表达。免疫组化和qRT-PCR分别检测Ref-1在肝癌组织和肝癌细胞系中表达;最后HepG2细胞同时转染了miR-mimics和Ref-1 miR-inhibitors,进行联合干预,随后CCK-8、细胞划痕和Transwell分别检测了联合干预后细胞增殖、迁移和侵袭能力变化。结果:miR-133a-3p在肝癌患者癌组织中的表达显著低于癌旁组织,同时其在肝癌细胞HepG2中的表达最低;qRT-PCR结果表明miR-mimics可有效促进miR-133a-3p在HepG2中的表达;CCK-8和克隆形成实验结果显示miR-133a-3p过表达后可以显著抑制HepG2细胞的增殖、迁移和侵袭等恶性生物学行为;RNA-seq结果显示HepG2转染前后Ref-1的表达差异最大;qRT-PCR和Western blot结果显示HepG2转染成功后,Ref-1表达随miR-133a-3p的过表达而降低。免疫组化和qRT-PCR结果显示Ref-1在肝癌组织和肝癌细胞系中的表达均显著高于阴性对照组;联合干预后HepG2细胞的增殖、迁移和侵袭等恶性生物学抑制作用更加强烈。结论:miR-133a-3p可能通过抑制Ref-1表达从而抑制肝癌细胞的增殖、迁移和侵袭等恶性生物学行为。     

涎腺腺样囊性癌中Cyclin D1、MMP-7和PRB2/p130表达及意义

目的探讨Cyclin D1、MMP-7和PRB2/p130蛋白在涎腺腺样囊性癌组织中的表达及与涎腺腺样囊性癌生物学行为的关系。方法应用免疫组化SP法检测46例涎腺腺样囊性癌和30例正常涎腺组织中Cyclin D1、MMP-7和PRB2/p130蛋白的表达。结果 Cyclin D1、MMP-7和PRB2/p130在涎腺腺样囊性癌中的阳性率分别为71.74%、67.39%、63.04%;Cyclin D1和PRB2/p130蛋白在涎腺腺样囊性癌中的表达与病理类型有关(P<0.05),MMP-7在涎腺腺样囊性癌中的表达与病理类型无关(P>0.05);Cyclin D1、MMP-7和PRB2/p130蛋白在涎腺腺样囊性癌复发组中的阳性率与无复发组相比,差异有显著性(P<0.05);Cyclin D1与PRB2/p130在涎腺腺样囊性癌中的表达呈负相关,与MMP-7表达呈正相关。结论 Cyclin D1、MMP-7和PRB2/p130蛋白的异常表达在涎腺腺样囊性癌发生、发展和复发中起重要作用,联合检测多个指标可能对患者预后的判断有指导意义。     

TRIM11靶向调控miR-24-3p促进乳腺癌细胞系MCF7增殖和侵袭

目的探讨TRIM11靶向调控miR-24-3p对乳腺癌细胞增殖和侵袭的影响,明确TRIM11高表达与乳腺癌预后的相关性。方法免疫组化法检测TRIM11在31例乳腺癌和31例癌旁正常组织中的表达。用siRNA TRIM11和miR-24-3p慢病毒转染人乳腺癌MCF7细胞,实时荧光定量PCR(RT-q PCR)和Western bolt检测TRIM11、miR-24-3p mRNA和蛋白表达变化及相关性分析;荧光素酶报告实验验证miR-24-3p为TRIM11的潜在靶点;MTT法和Transwell检测细胞增殖数量、增殖活力和侵袭。结果 TRIM11在乳腺癌组织及MCF7细胞中表达显著增高(P0.05)。TRIM11高表达患者生存率显著降低(P0.05)。siRNA TRIM11转染显著抑制MCF7细胞中TRIM11表达,而且抑制MCF7细胞增殖数量、增殖活力和侵袭能力(P0.05)。miR-24-3p显著降低野生型3'-UTR TRIM11荧光素酶活性(P0.05),但对突变型没有作用。miR-24-3p mRNA在MCF7细胞下调,miR-24-3p抑制MCF7细胞TRIM11蛋白和mRNA表达,miR-24-3p mRNA与TRIM11 mRNA在MCF7细胞中表达显著负相关(P0.05),miR-24-3p抑制MCF7细胞增殖数量、增殖活力和侵袭能力(P0.05)。结论 TRIM11在乳腺癌组织及MCF7细胞中表达上调,并可能通过靶向调控miR-24-3p促进乳腺癌细胞增殖和侵袭,进而影响乳腺癌预后,TRIM11/miR-24-3p轴有望成为治疗乳腺癌的新靶点。     

力学刺激对腺样囊性癌细胞株侵袭力的影响

我们旨在观察周期性双轴力学刺激对腺样囊性癌高转移细胞株ACC-M,腺样囊性癌低转移细胞株ACC-2体外侵袭力以及MMP-9表达的影响,以探讨腺样囊性癌的侵袭机制。分别选取4000微应变和1000微应变加载,将ACC-M,ACC-2每天加载1次,加载时间分别为1、3和6h,共加载2d。以在相同培养小室中静态培养组作为对照。细胞在力学刺激后行MMP-9免疫荧光染色,用激光共聚焦显微镜和图像分析软件进行定量分析。用Transwell小室行ACC-M,ACC-2细胞体外侵袭力测定。结果表明:MMP-9在ACC-M细胞表达明显高于ACC-2细胞,提示MMP-9与ACC的侵袭有密切关系。在力学刺激下,ACC-M和ACC-2细胞的MMP-9的表达及侵袭能力随力学刺激的大小和作用时间的变化而变化。ACC-M,ACC-2细胞在力学刺激后出现侵袭能力的改变,同时还可改变其基质金属蛋白酶-9的表达,显示力学刺激可以调节肿瘤转移,我们认为有基质金属蛋白酶-9之外的蛋白在其中起作用。     

miR-210调控Mex3B蛋白对肺癌细胞侵袭和增殖的影响

目的:探讨miR-210和Mex3B蛋白对肺癌细胞侵袭和增殖能力的影响。方法:运用q PCR检测miR-210在正常肺组织和癌旁组织中的表达情况;运用q PCR检测Mex3B在肺癌组织、癌旁组织中的表达;q PCR检测miR-210在不同肺癌细胞株中(A549、H1299、H1650和H358)的表达水平;双荧光素酶报告基因系统检测miR-210对Mex3B转录的影响;细胞活性实验检测miR-210的表达对肺癌细胞活性的影响;平板克隆实验检测miR-210的表达对肺癌细胞A549的增殖能力的影响;Transwell侵袭实验检测miR-210的表达对肺癌细胞株A549的侵袭能力的影响。结果:和癌旁组织比较,miR-210在肺癌组织中表达明显增高,和癌旁组织比较,Mex3B在肺癌中表达较低,双荧光素酶报告基因系统检测结果显示,miR-210可以直接调控Mex3B的转录活性,抑制miR-210的表达活性后,A549肺癌细胞的细胞活性受到一定的抑制;抑制miR-210后,肺癌细胞株A549的侵袭和增殖能力明显降低。结论:miR-210可以靶向调控肺癌细胞的侵袭和增殖能力。     

Modes of reinforcement and gender and culture of experimenter and subject: effects of "alien" and external reinforcement for Japanese and American men and women

In this study, we examined external and "alien" reinforcement (ER and AR. respectively) as a factor in social learning, and studied the combined effects of culture and reinforcement mode. A female (Experiment 1) and a male (Experiment 2) experimenters conducted experimental sessions. Both men and women, who grew up in the same culture as the experimenter, participated and performed the experimental task. A three-way interaction effect of experimenter gender, culture, and reinforcement mode was found on task performance. And the effect was more pronounced for a Japanese experimenter. A female and a male experimenters conducted Experiments 3 and 4, respectively; however participants this time were men and women who grew up in different cultures than the experimenter. Results indicated that the pattern of the subject gender and reinforcement mode interaction effect, when the experimenter was Japanese with American subjects, was exactly opposite to that when the experimenter was American. These experiments showed that AR was as effective for social learning as ER, and that the cultural backgrounds of experimenter and subject influenced AR and ER effectiveness.     

Timing and intensity of early fevers and the development of allergies and asthma

BACKGROUND: Early childhood fevers appear to protect against later allergies and asthma. What is not known is the time in which fevers exert this effect and whether the degree of temperature increase is important. OBJECTIVE: We sought to examine the relationship between the time and degree of early fevers and later allergies and asthma. METHODS: Eight hundred thirty-five children from southeast Michigan were enrolled at birth. Clinic records from their first 2 years were abstracted for episodes of fever. At age 6 to 7 years, children underwent allergy testing. We examined fevers occurring within 6-month intervals in the first 2 years of life and outcomes at age 6 to 7 years. The primary outcome measures were allergic sensitization, asthma, asthma with allergic sensitization, and asthma without allergic sensitization. RESULTS: In the unadjusted analysis each episode of fever between 7 and 12 months of age was associated with a lower odds of allergic sensitization (odds ratio [OR], 0.71; 95% CI, 0.54-0.93) and asthma with allergic sensitization (OR, 0.43; 95% CI, 0.21-0.90) at age 6 to 7 years. Likewise, every 1 degrees C increase in the maximum temperature between 7 and 12 months was associated with a lower odds of allergic sensitization (OR, 0.77; 95% CI, 0.61-0.96) and asthma with allergic sensitization (OR, 0.62; 95% CI, 0.40-0.94). After adjusting for potential confounders, each episode of fever between 7 and 12 months was associated with a lower likelihood of asthma with allergic sensitization (adjusted OR, 0.33; 95% CI, 0.11-0.94) at age 6 to 7 years. CONCLUSIONS: Both the timing and intensity of childhood fevers appear to be important factors in the development of allergies and asthma.     

Rates of oxygen consumption and of anaerobic glycolysis in renal cortex and medulla of adult and new-born rats and guinea-pigs

1. Rates of oxygen uptake and of anaerobic glycolysis were estimated in slices from the renal cortex and medulla (a) of adult rats and guinea-pigs, (b) of new-born (1-, 5- and 21-day-old) rats and of guinea-pigs of 1, 12, 21, 24 and 120 hr age.2. In the adult rat, Q(O2) values for the cortex were 12.55 +/- 0.20 (22) and for the medulla: 8.56 +/- 0.17 (22) mul./hr.mg dry weight, while in the new-born rat (24 hr old) they were 10.99 +/- 0.46 (12) and 9.33 +/- 0.18 (9) mul./hr.mg dry weight respectively.3. Values for Q(CO2) (N2) (anaerobic glycolysis) in the 14 hr old new-born rat were in the renal cortex 9.65 +/- 0.35 (5) and in the medulla 7.39 +/- 0.43 (5) mul./hr.mg dry weight; while in the adult they were 2.25 +/- 0.08 (16) and 5.76 +/- 0.14 (16) mul./hr.mg dry weight, respectively.4. In the adult guinea-pig values for Q(CO2) (N2) were of the same order as in the adult rat, though the rate of O(2) uptake was for the cortex 8.12 +/- 0.22 (12) and for the medulla 5.02 +/- 0.23 (11) mul./hr.mg dry weight.5. Though the Q(O2) values in the renal cortex and medulla were smaller in the 1 hr old new-born guinea-pig, they were already increasing in the 12 hr old neonate.6. The results are discussed in the light of enzyme changes occurring during the process of maturation of the nephron as indicated by histochemical observations.     

Association between functional alterations of senescence and senility and disorders of gait and balance

OBJECTIVES:

Declines in cognition and mobility are frequently observed in the elderly, and it has been suggested that the appearance of gait disorders in older individuals may constitute a marker of cognitive decline that precedes significant findings in functional performance screening tests. This study sought to evaluate the relationship between functional capacities and gait and balance in an elderly community monitored by the Preventive and Integrated Care Unit of the Hospital Adventista Silvestre in Rio de Janeiro, RJ, Brazil.

METHODS:

Elderly individuals (193 females and 90 males) were submitted to a broad geriatric evaluation, which included the following tests: 1) a performance-oriented mobility assessment (POMA) to evaluate gait; 2) a mini-mental state examination (MMSE); 3) the use of Katz and Lawton scales to assess functional capacity; 4) the application of the geriatric depression scale (GDS); and 5) a mini-nutritional assessment (MNA) scale.

RESULTS:

Reductions in MMSE, Katz and Lawton scores were associated with reductions in POMA scores, and we also observed that significant reductions in POMA scores were present in persons for whom the MMSE and Katz scores did not clearly indicate cognitive dysfunction. We also demonstrated that a decline in the scores obtained with the GDS and MNA scales was associated with a decline in the POMA scores.

CONCLUSIONS:

Considering that significant alterations in the POMA scores were observed prior to the identification of significant alterations in cognitive capacity using either the MMSE or the Katz systems, a prospective study seems warranted to assess the predictive capacity of POMA scores regarding the associated decline in functional capacity.     

男性乳腺发育症临床病理分析及雌,孕激素受体检测

对１１３例男性乳腺发育症进行临床病理分析。同时检测其中３０例乳腺组织中雌激素受体和孕激素受体分布情况，结果发现两者阳性率分别为８０．０％和８３．３３％。结合文献讨论了男性乳腺发育症的发生与高血清激素浓度及乳腺组织高受体水平的关系。