雌激素及其受体在子宫内膜息肉发生、发展中的作用机制研究

目的 探讨雌激素及其受体(ERβ)在子宫内膜息肉发生、发展中的作用机制.方法 选择子宫内膜息肉患者45例作为子宫内膜息肉组;另外选择同期正常子宫内膜组织20例作为正常子宫内膜组.检测雌二醇的浓度、雌性激素受体α、β(ERα、ERβ)的含量并比较.结果 子宫内膜息肉组患者子宫内膜组织中雌二醇浓度为(2.8±0.9) pmol/L,外周血中雌二醇浓度为(68.2±10.7) poml/L,两者呈正相关关系(r=0.786,P＜0.01);正常子宫内膜组患者子宫内膜组织中雌二醇浓度为(2.9 ±0.1) pmol/L,外周血中雌二醇浓度为(66.9±14.5) pmol/L,两者亦呈正相关关系(r=0.790,P＜0.01);子宫内膜息肉组和正常子宫内膜组中子宫内膜间质细胞中的ERα、ERβ含量差异均有统计学意义(均P＜0.05).结论 雌激素及其受体在子宫内膜间质细胞中的高表达,可能是子宫内膜息肉的发生重要因素.     

子宫内膜异位症痛经患者内膜组织中雌激素受体表达的相关研究

目的 检测内异症痛经患者内膜组织中雌激素受体的表达,分析其与疼痛及其程度的关系.方法 内异症组50例,其中在位内膜组40例,异位内膜组10例,对照组30例,在位内膜组患者术前均对痛经程度进行评分,采用免疫组化法测定雌激素受体的表达.结果 EMs组在位内膜ER蛋白的表达高于对照组,差异有显著性(P＜0.05);EMs疼痛组与无痛组的表达差异有显著性(P＜0.05);ER蛋白在EMs重度疼痛组组织中阳性灰度值最高,而在轻度疼痛组灰度值最低,三组间比较差异均有显著性(P＜0.05).结论 内异症病灶局部雌激素受体表达增加,可能与子宫内膜异位症疼痛的发生、发展密切相关.     

人雌激素受体β绿色荧光蛋白真核表达载体的构建与表达

目的构建人雌激素受体β(ERβ)绿色荧光蛋白(GFP)真核表达载体,并稳定转染乳腺癌MCF-7细胞。方法通过双酶切将pCMV5-hERβ中的人ERβcDNA克隆到pEGFP-C3中,构建并鉴定重组质粒pEGFP-hERβ,然后转染乳腺癌MCF-7细胞,采用G418(500μg/mL)筛选出稳定转染的细胞系,在荧光显微镜下观察融合蛋白质在真核细胞内的表达分布,并采用逆转录-聚合酶链反应(RT-PCR)检测ERβ的mRNA转录水平。结果成功构建重组表达载体pEGFP-hERβ,有效转染并筛选出稳定转染ERβ基因的MCF-7细胞系,外源性ERβ基因在MCF-7细胞中获得了有效转录,并在细胞质和细胞核中均广泛分布。结论建立了稳定表达ERβ的MCF-7细胞系,为筛选ERβ调节剂和进一步研究ERβ在乳腺癌中的作用及其机制奠定基础。     

脂肪细胞雌激素受体的测定及表达水平的研究

目的　研究不同部位的脂肪细胞雌激素受体 (estrogenreceptor ,ER)的表达水平 ,探讨脂肪细胞与雌激素之间的关系。方法　对 64例成人腹部皮下脂肪组织及内脏大网膜脂肪组织 ,应用免疫组织化学法染色并进行图象分析。结果　成人脂肪细胞核内雌激素受体呈阳性反应 ;男性腹部皮下脂肪细胞的ER阳性细胞的光密度高于大网膜脂肪ER阳性细胞的光密度 (P <0 .0 5 ) ;女性的腹部皮下脂肪细胞的ER阳性细胞的光密度与大网膜脂肪组织ER阳性细胞的光密度相比差异无显著性 (P >0 .0 5 )。结论　 ( 1)脂肪细胞的ER免疫阳性反应表达于脂肪细胞的细胞核。 ( 2 )男性的不同部位脂肪细胞的ER表达不同 ,而女性不同部位脂肪细胞ER的表达一致     

KGF对无排卵型功血子宫内膜上皮细胞ER和PR的调节研究

目的:观察角质细胞生长因子(KGF)对离体培养的无排卵型功血子宫内膜上皮细胞(EEC)中雌激素受体(ER)和孕激素受体(PR)表达的影响.方法:体外培养无排卵型功血子宫内膜上皮细胞,加入不同浓度的角质细胞生长因子作用后,RT-PCR检测ER和PR的mRNA表达改变,并与在体无排卵型功血子宫内膜组织及培养的正常子宫内膜上皮细胞进行比较.结果:RT-PCR方法检测两组ER和PR的mRNA的表达情况,结果显示功血组较对照组均显著增高(P<0.05).结论:KGF对体外培养的EEC有增加ER和PR的作用,但程度不如正常的子宫内膜上皮细胞,提示无排卵功血可能存在与正常的子宫内膜上皮细胞不同的生长机制,但二者的作用均受KGF的影响.     

子宫内膜增生组织中Cath-D、PCNA、ER和PR的表达

目的 探讨组织蛋白酶D(Cath-D)、增殖细胞核抗原(PCNA)、雌激素受体(ER)和孕激素受体(PR)在不同类型子宫内膜增生组织中的表达.方法 60例子宫内膜增生患者分为不典型增生(16例,AH组)、复杂性增生(18例,CH组)和单纯性增生(26例,SH组);另选取20例正常的增殖期子宫内膜作为对照组.用免疫组化法检测增生子宫内膜组织Cath-D、PCNA、ER和PR的阳性表达率.结果 四组ER的阳性表达率相仿(P＞0.05).AH组、CH组增生子宫内膜组织中的PR表达均明显低于对照组(P＜0.05);AH组和CH组的Cath-D和PCNA的阳性表达率均明显高于对照组(P＜0.01);SH组、CH组、AH组细胞核中Cath-D、PCNA的阳性表达率逐级递增,而PR的表达水平逐渐减弱.结论 检测子宫内膜增生组织中Cath-D、PCNA、PR的表达水平有助于评估子宫内膜增生的预后,指导临床治疗.     

异常子宫出血患者单纯增生型与正常增殖期子宫内膜ER、PR表达研究

目的 观察异常子宫出血患者单纯增型与正常增生期子宫内膜雌激素受体（Estrogen receptor，ER）、孕激素受体（Progestron receptor，PR）表达有无差异。方法 门诊宫腔镜下观察需行诊刮的异常子宫出血患者的子宫内膜组织，石蜡包埋制成病理切片，HE染色按内膜的病理类型分为单纯增生型和正常增生期子宫内膜两组，用免疫组织化学方法测定ER、PR含量，采用LeicaQ500IW图像分析仪进行测定，应用光密度值进行比较。结果 异常子宫出血患者单纯增生型和正常增生期子宫内膜的ER、PR分布不同，单纯增生病变部位ER、PR含量高于正子宫内膜组织部位。结论 异常子宫出血患者单纯增生型和正常增殖期子宫内膜的ER、PR含量不同；ER、PR在子宫内膜增殖中起了非常重要的作用。     

异常子宫出血患者子宫内膜中ER、PR分布情况研究

目的 观察异常子宫出血患者子宫腔内各部位内膜组织雌激素受体(Estrogen receptor,ER)、孕激素受体(Progestron receptor,PR)表达有无差异,为宫腔镜下子宫内膜电切术切除范围提供理论依据.方法 门诊宫腔镜下观察异常子宫出血须行诊刮及部分行子宫全切除术的患者宫腔内不同部位的子宫内膜组织,用免疫组织化学方法测定ER、PR含量,采用LeicaQ500IW图像分析仪进行测定,应用光密度值进行比较.结果 异常子宫出血患者子宫腔内各部位内膜增生情况不同;ER、PR含量从高到低的顺序为:宫底＞子宫体部＞子宫角部＞宫颈峡部;ER、PR在子宫腔内各部位分布情况的不同具有相关性.结论 对于宫底部高ER、PR状态,应引起高度重视;经宫颈子宫内膜电切术(transcervical resection of endometrium,TCRE)或子宫内膜热球剥脱术(uterine balloon thermablate,UBT).术中谨慎并彻底地处理宫底与双侧宫角部的子宫内膜组织,破坏其内膜的结构,避免复发和恶变.     

E0703对雌激素受体(ER)不同亚型的选择性激活作用

目的探讨E0703(雌二醇衍生物)对雌激素受体不同亚型(α和β)的选择性激活作用。方法将报告基因质粒pTAL-SEAP-ERE和校正质粒pSV-β-gal共转染已稳定转染了雌激素受体α/β的人胚肾细胞(HEK293),加入E0703进行报告基因的诱导表达。采用EMSA试验观察E0703对ERα与雌激素反应元件(ERE)的结合增强作用。结果E0703对ERα具有较强的激活作用(最低激活浓度为10-8mol·L-1),激活强度略低于E2;E0703在10-10、10-9、10-8、10-7mol·L-1浓度下对ERβ没有激活作用;EMSA试验证明,E0703能够增强ERα与雌激素反应元件的结合增强作用。结论E0703对雌激素受体α具有选择性激活作用。     

分析雌激素受体(ER)与IP3R在子宫肌瘤细胞中的表达价值

目的分析雌激素受体(ER)与IP3R在子宫肌瘤细胞中的表达价值。方法选择2012年12月至2013年5月于我院治疗的40例子宫肌瘤患者为实验组,并以同期于我院治疗的40例非子宫肌瘤患者作对照组进行观察。比较两组患者治疗前后的雌激素受体(ER)和IP3R水平。结果治疗前对照组患者体内雌激素受体(ER)、IP3R水平明显优于实验组组(P<0.05);手术治疗后实验组患者雌激素受体(ER)、IP3R水平有所好转,但仍比对照组差(P<0.05)。结论子宫肌瘤细胞中雌激素受体(ER)与IP3R在子宫肌瘤的临床中具有特异型,值得临床进一步研究。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.