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1.
逆转录多聚酶链反应方法研究乳腺癌的微小转换   总被引:3,自引:1,他引:2  
目的 通过分子生物学与常规免疫组化法比较,以寻求更好地检测乳腺癌微小转移的方法。方法 采用逆转录多聚酶链反应(RT-PCR)方法检测26例乳腺癌患者的外周血与骨髓中细胞角蛋白19(KT19)mRNA的表达,并用链酶亲和素-生物素-过氧化物酶复合物(SABC)免疫组化法检测乳腺癌患者骨髓涂片中上皮膜抗原(EMA)。结果 26例外周血中KT19 mRNA阳性4例(15.4%),骨髓阳性10例(38.4%)。免疫组化结果显示26例骨髓中有7例(26.9%)EMA阳性,其KT19mRNA都阳性,有3例(11.5%)免疫组化结果阴性而T19mRNA阳性。结论 RT-PCR方法检测KT19mRNA是一种比较敏感的方法,免疫组化也是一种比较可靠的检测方法,外周血中KT19mRNA的检测结果还不能完全取代骨髓的微小转移情况。  相似文献   

2.
Normal squamous epithelial cells readily undergo terminal differentiation in culture and are commonly used in differentiation studies. Several intracellular markers of squamous differentiation such as keratin, involucrin, transglutaminase and cholesterol sulfate have been well-studied and described by other workers. We have recently reported a surface membrane antigen in squamous carcinoma of the head and neck antigen in squamous carcinoma of the head and neck which is recognized by a murine monoclonal antibody SQMI. In this paper, we present our studies on the ultrastructural localization of SQMI antigen in cultured squamous epithelial cells using gold-labelled antibody. The cells studied included both normal and cancer cells at different degrees of differentiation. Under both transmission and scanning electron microscopy examination, the SQMI antigen was localized at the membrane surface of cultured cells, particularly at sites of cell-cell interdigitation. No association with desmosomal structure was observed in any of the specimens examined. There was however an association of SQMI antigen with microvilli of cell membrane. No non-specific cytoplasmic localization of SQMI antigen was observed. The intensity of SQM1 antigen revealed by gold-labelling appeared to have a positive correlation with the degree of differentiation of the cells in culture.  相似文献   

3.
目的 深入了解卵巢交界性子宫内膜样肿瘤(EBT)的临床病理特点、鉴别诊断、治疗和预后.方法 对清远市人民医院病理科2012年1月至2019年9月诊断的4例卵巢EBT的临床资料、组织学形态、免疫组化特点及预后进行分析总结,并复习相关文献.结果 4例因不规则阴道出血、下腹胀痛或发现盆腔占位入院,均行子宫及双附件切除术,其中...  相似文献   

4.
The effect of saiboku-to on gastric lesions induced by restraint water-immersion stress and ethanol has been examined in rats. Thirty minutes after oral administration of saiboku-to, the rats were placed in restraint cages and immersed in water at 23 degrees C for 7 h, or orally administered 99.5% ethanol (1 mL) and placed in normal cages for 1 h. The stress for 7 h or the ethanol treatment for 1h induced erosion in the glandular area of the stomach. Histology showed that the surface epithelial cells were desquamated and part of the lamina propria mucosae was injured. The evaluation of lesion index, the cumulative length of the gastric lesion, on the gross appearance of the stomach, revealed that saiboku-to dose-dependently inhibited both the water-immersion stress-induced gastric erosion and ethanol-induced gastric erosion. To determine whether the anti-erosion effect of saiboku-to was because of a mild irritant effect, saiboku-to or 20% ethanol, which is known as a typical mild irritant, were given orally. After 30 min a strong irritant, 99.5% ethanol, was given orally. Histological examination was performed 30 min after administration of saiboku-to or the mild irritant, and 1 h after administration of the strong irritant. The mild irritant induced a reduction in surface epithelial cells 30 min after administration. Furthermore, the mild irritant protected the stomach against mucosal erosion produced by the strong irritant. Saiboku-to protected the strong irritant-induced erosion without producing mild irritation as observed in stomach treated with 20% ethanol. Pretreatment with saiboku-to also inhibited the decrease in the levels of hexosamine, gastric mucus glycoprotein, induced by the strong irritant. In pylorus-ligated rats, saiboku-to dose-dependently inhibited gastric acid secretion, a gastric aggressive factor. These results suggest that the anti-erosion effect of saiboku-to which is not a mild irritant, involves both inhibition of aggressive factors, such as gastric acid secretion, and augmentation of defensive factors, such as gastric mucus cells.  相似文献   

5.
目的 探讨外周血单核细胞作为角膜缘干细胞替代角膜上皮细胞再生的潜能。方法 实验时间为2020年1月至2021年1月,实验动物为来自暨南大学生理实验室5~8月龄健康新西兰大白兔30只;外周血单核细胞体外培养及鉴定:采用密度梯度离心法分离兔外周血单核细胞,在倒置相差显微镜下观察单核细胞形态,细胞分选技术检测单核细胞表面标志物CD14。将收集的单核迁移细胞置于新鲜角膜缘和角膜上皮组织块的微环境中共培养1周,流式细胞仪检测诱导前与诱导后细胞表面特异标志物细胞角蛋白的阳性率,诱导前与诱导后细胞表面角蛋白阳性表达率指数的组间比较采用重复测量设计的方差分析。计量资料采用t检验。结果 流式细胞术检测诱导前单核迁移细胞特异标志物细胞角蛋白阳性率为1.64%,诱导后单核迁移细胞特异标志物细胞角蛋白阳性率为11.30%。诱导前,实验组细胞角蛋白阳性表达率与对照组比较,差异无统计学意义(P>0.05);诱导7 d后,实验组细胞角蛋白阳性表达率为(10.298±0.996)%,与对照组(1.786±0.237)%比较,差异有统计学意义(P<0.05);实验组诱导前细胞角蛋白阳性表达率为(1.308±0.376)%,与诱导7 d后(10.298±0.996)%比较,差异有统计学意义(P<0.05);对照组诱导前细胞角蛋白阳性表达率与诱导7 d后比较,差异无统计学意义(P>0.05)。结论 外周血单核细胞作为角膜缘干细胞的自体替代干细胞来源,被认为是角膜上皮重建的有效治疗策略。  相似文献   

6.
Metastasis is one of hallmarks of cancer and a major cause of cancer death. Combatting metastasis is highly challenging. To overcome these difficulties, researchers have focused on physical properties of metastatic cancer cells. Metastatic cancer cells from patients are softer than benign cancer or normal cells. Changes of viscoelasticity of cancer cells are related to the keratin network. Unexpectedly, keratin network is dynamic and regulation of keratin network is important to the metastasis of cancer. Keratin is composed of heteropolymer of type I and II. Keratin connects from the plasma membrane to nucleus. Several proteins including kinases, and protein phosphatases bind to keratin intermediate filaments. Several endogenous compounds or toxic compounds induce phosphorylation and reorganization of keratin network in cancer cells, leading to increased migration. Continuous phosphorylation of keratin results in loss of keratin, which is one of the features of epithelial mesenchymal transition (EMT). Therefore, several proteins involved in phosphorylation and reorganization of keratin also have a role in EMT. It is likely that compounds controlling phosphorylation and reorganization of keratin are potential candidates for combating EMT and metastasis.  相似文献   

7.
Localization of N-acetyltransferases NAT1 and NAT2 in human tissues.   总被引:5,自引:0,他引:5  
Human acetyl coenzyme A-dependent N-acetyltransferase (EC 2.3.1.5) (NAT) catalyzes the biotransformation of a number of arylamine and hydrazine compounds. NAT isozymes are encoded at 2 loci; one encodes NAT1, formerly known as the monomorphic form of the enzyme, while the other encodes the polymorphic NAT2, which is responsible for individual differences in the ability to acetylate certain compounds. Human epidemiological studies have suggested an association between the "acetylator phenotype" and particular cancers such as those of the bladder and colon. In the present study, NAT1- and NAT2-specific riboprobes were used in hybridization histochemistry studies to localize NAT1 and NAT2 mRNA sequences in formalin-fixed, paraffin-embedded human tissue sections. Expression of both NAT1 and NAT2 mRNA was observed in liver, gastrointestinal tract tissues (esophagus, stomach, small intestine, and colon), ureter, bladder, and lung. In extrahepatic tissues, NAT1 and NAT2 mRNA expression was localized to intestinal epithelial cells, urothelial cells, and the epithelial cells of the respiratory bronchioles. The observed heterogeneity of NAT1 and NAT2 mRNA expression between human tissue types may be of significance in assessing their contribution to known organ-specific toxicities of various arylamine drugs and carcinogens.  相似文献   

8.
The differential diagnosis of submucosal stomach lesions includes gastrointestinal stromal tumour (GIST), leiomyoma, synovial sarcomas, perineurioma, myxoid chondrosarcoma, myoepithelial tumour and other rare mesenchymal tumours. GISTs are well-defined lesions with distinctive morphologic and histogenetic characteristics that show 95% positive staining for CD117. Differential diagnosis of wild-type GISTs can be challenging. Here, we present two stomach tumours that were operated on in our surgical department. Both presented with positive immunoreactivity for CD117. In one tumour, c-Kit mutation analysis demonstrated positivity of exon 11_c.1674_1676delGGT, thus confirming the diagnosis of a GIST. Mutational analysis of the second stomach lesion demonstrated negativity for all known c-KIT and PDGFRA exons. In situ hybridisation ruled out a synovial sarcoma. An additional immunohistochemical staining for epithelial membrane antigen eventually confirmed the diagnosis of an extremely rare reticular perineurioma in the stomach, so far reported for the second time worldwide. Both patients have not shown any signs of recurrence 2 years after surgery. The presented cases emphasise the benefits of performing a mutational analysis in difficult GISTs, including wt-GISTs, and demonstrates the importance and challenges in differentiating GISTs from other mesenchymal tumours.  相似文献   

9.
Ten cases of ameloblastoma including a case of gingival origin and a case of atypical ameloblastoma were studied immunohistochemically, especially with monoclonal and polyclonal anti-keratin antibodies. Stellate reticulum cells reacted strongly with monoclonal and polyclonal anti-keratin antibodies but columnar cells were reacted weakly. The level of keratinization of columnar cells was similar to that of the basal cell layer of the gingival squamous epithelium. Our immunohistochemical studies suggested that the process of keratinization-ameloblastoma follows the sequence of 1) cuboidal cells 2) columnar cells 3) stellate reticulum cells and 4) metaplastic squamous cells. The findings also disclosed the epithelial nature of granular cells. The reactivity of the components of atypical ameloblastoma differed from that of other cases. Other positive "markers" in ameloblastomas included CEA and CA19-9, but no particular histologic differences were observed between positive and negative cases.  相似文献   

10.
目的:探讨热休克蛋白70(heat shock protein70,HSP70)在老年性白内障晶状体上皮细胞中的表达及意义.方法: 采用PV-6000通用型免疫组织化学染色,检测老年性白内障晶状体前囊膜53例,其中皮质性白内障24例,核性白内障17例,后囊下白内障12例,正常人晶状体前囊膜6例中晶状体上皮细胞中HSP70的表达.并进行阳性细胞计数. 结果:老年性白内障与正常晶状体上皮细胞中均有HSP70表达,其中HSP70在老年性白内障中表达明显高于正常对照组,差异有显著性意义(P<0.05).老年性白内障组各亚型间有显著性差异(P<0.05).结论:HSP70可能在老年性白内障的发生、发展中起重要作用.  相似文献   

11.

Background and purpose:

Mucosal microcirculation is compromised during gastric damage induced by non-steroidal anti-inflammatory drugs, such as aspirin. Consequently, oxygen supply to epithelial cells is decreased. The trefoil factor (TFF) peptides are involved in mechanisms of defence and repair in the gastrointestinal tract but their regulation at sites of gastric injury is unknown.

Experimental approach:

Hypoxia and expression of TFF genes and peptides were measured in the damaged stomach of aspirin-treated rats. In a human gastric cell line (AGS cells), the effects of hypoxia and of hypoxia inducible factor (HIF)-1 (through transient transfection of HIF-1α siRNA or over-expression of HIF-1α) on TFF gene expression were evaluated.

Key results:

Hypoxyprobe immunostaining, up-regulation of TFF2 (1.9-fold) and TFF3 (1.8-fold) and a non-significant increase of TFF1 (1.5-fold) mRNA were observed in the damaged stomach of aspirin-treated rats, compared with control animals. Hypoxia (3% O2, 16 h) induced mRNA for TFF1 (5.8-fold), TTF2 (9.1-fold) and TFF3 (9.3-fold) in AGS cells, an effect mediated by HIF-1, as transient transfection of HIF-1α siRNA reduced the effects of hypoxia. Over-expression of HIF-1α by transfection in non-hypoxic epithelial cells produced a similar pattern of TFF induction to that observed with hypoxia and transactivated a TFF1 reporter construct.

Conclusions and implications:

Hypoxia inducible factor-1 mediated the induction of TFF gene expression by hypoxia in gastric epithelial cells. Low oxygen levels and up-regulation of TFF gene expression in the damaged stomach of aspirin-treated rats suggest that hypoxia induced expression of TFF genes at sites of gastric injury.  相似文献   

12.
To investigated spontaneous benign prostatic hyperplasia (BPH) in dog the effect of a synthetic steroidal antiandrogen, chlormadinone acetate (CMA) was studied. Old male beagle dogs (5-8 years old) were divided into following experimental groups: group 1 consisted of BPH controls; group 2 received CMA 0.3mg/kg/day p.o., for 6 months. In group 1 animals, glandular hyperplasia of the prostate was clearly detected. The glandular epithelial cells showed uniformly intense immunostaining for nuclear androgen receptors (AR). AR was also localized in the nuclei of the fibro-muscular cells. Immunoreactivity of 5alpha-reductase type I was positive in most glandular epithelial cells. The staining was positive in the cytoplasm but not in the nuclei. No fibro-muscular cells were stained. In contrast, CMA produced marked atrophy of the glandular epithelium. The interacinar fibro-muscular stroma was prominent. Furthermore, immunostaining of nuclear AR of both epithelial and stroma cells was remarkably decreased. The intensity of staining for 5alpha-reductase type I in most glandular epithelial cells also decreased. Interestingly, some basal cells exhibited positive staining for 5alpha-reductase type I. These results indicate that the uptake of testosterone and/or its androgenic effect on the prostate may be suppressed by CMA. We further speculate that the basal cells produce sufficient dihydrotestosterone to maintain themselves even in the presence of low testosterone levels.  相似文献   

13.
Congenital defects of predominantly cell-mediated immunity without other major defects or recognizable enzyme deficiency are classified "severe combined immunodeficiency" (SCID). Affected thymuses are severely reduced in size, microscopically they show only few lymphocytic cells and a cortico-medullary boundary is missing as well as Hassal's corpuscules. The primary defect is not yet clear. In this study 4 such cases were examined. The intrathymic epithelial cells showed expression of different cytokeratins and the typical strong positivity for the major histocompatibility complex. Lymphoid cells were phenotyped immunohistochemically and appeared as a very small population of cortical thymocytes, which was especially clear in a prenatally diagnosed and aborted fetus. His thymus, not altered by stress of infection or bone marrow transplantation, showed those few lymphocytes clustered around epithelial cells. The finding suggests an arrest in maturation and cell amplification rather than the absence of stem cells in the pathogenesis of SCID. Alterations in the thymic microenvironment could not be revealed by our methods.  相似文献   

14.
脂肪干细胞复合多孔丝素材料修复兔尿道缺损   总被引:3,自引:1,他引:2  
目的 探索脂肪间充质干细胞(ADMSCs)复合多孔丝素膜(PSFS)对尿道缺损的修复效果.方法 建立雄性新西兰大白兔尿道缺损模型后分为3组,每组13只.A组用PSFS修补,B组用BrdU标记ADMSCs复合PSFS修补,C组为尿道缺损对照.手术前后行尿道造影.术后2、4、6周取修复段尿道行组织学检查,免疫组织化学染色检测BrdU标记ADMSCs、广谱角蛋白阳性细胞及巨噬细胞情况.结果 术后C组尿道狭窄和尿屡的并发症总发生率为76.92% ,高于A组的23.07% 和B组的15.38% (P<0.05).组织学观察C组的血管、上皮细胞层、平滑肌重建少;A、B组重建最多、B组多于A组.BrdU标记ADMSCs在3时间点缺损区丝素膜中散在可见;广谱角蛋白染色C组缺乏尿道上皮的复层柱状上皮形态,无乳突状黏膜存在;A、B组与正常尿道黏膜相似,有较多乳突状结构;术后4周巨噬细胞表达B组比A组少[(11.66±1.58)/HP vs.(13.88±2.08)/HP](P<0.05).结论 ADMSCs复合PSFS具有促进尿道缺损修复的作用.  相似文献   

15.
发生于胃的胃肠间质瘤外科治疗   总被引:1,自引:0,他引:1  
目的:探讨发生于胃的胃肠间质瘤(GIST)外科治疗的方法。方法:对1995年8月-2005年8月收治的经手术病理证实的19例胃GIST进行回顾性研究。结果:19例患者中,肿瘤位于贲门胃底区7例,胃体9例,胃窦3例。肿瘤最小2cm,最大25cm,中位大小6cm。胃周淋巴结转移1例。手术行胃楔形切除11例,近端胃大部切除5例,远端胃大部切除3例,其中联合脏器切除3例。查免疫组织化学CD117阳性16例,CD34阳性17例。术后随诊5例复发转移。结论:胃GIST治疗以手术为主,术中应根据肿瘤大小、部位、有无外侵及淋巴结转移决定手术范围,切除在于完全而不在于大范围清扫。  相似文献   

16.
Hyaluronic acid (hyaluronan, or HA) is secreted by submucosal glands, but its function in airway secretions other than influencing the rheology of mucus is not fully understood. HA is known to modulate cell behavior and to enhance sperm motility. Because sperm tails and cilia have the same microtubular structure, we studied the effect of HA on ciliary beat frequency (CBF) in vitro. CBF of cultured ovine airway epithelial cells was measured continuously by digital video microscopy. After removal of endogenous HA by hyaluronidase, cells were exposed to 50 to 100 microg/mL of HA at different times in culture. No change in CBF in response to HA was seen in cells cultured less than 7 days. After 7 days, however, 6 of 10 measured cells (from three different sheep) showed a transient CBF increase from a baseline of 6.4 +/- 0.3 Hz (mean +/- SE) to 7.4 +/- 0.4 Hz or 16% above baseline (p < 0.05). At these time points (but not before), cytochemical staining was positive for endogenous HA using a biotinylated HA-binding protein. These data suggest that HA can increase CBF of tracheal epithelial cells only late in culture when HA is able to bind to an unspecified cell surface structure. Because this binding has a physiological effect, we hypothesize that it is an HA-binding receptor, that is either transiently expressed late in culture or initially destroyed by the protease treatment for cell dispersion.  相似文献   

17.
【摘要】 目的 了解异位子宫内膜干细胞在病灶中的数量、分布及定位情况,并筛选其可能的标志物表达。方法 在子宫内膜异位症裸鼠模型建造前24h开始进行5-溴脱氧尿苷(BrdU)脉冲标记,共5d。在标记结束后4h、1周、3周、5周、6周、7周、8周及9周,通过免疫组化检测异位病灶中BrdU的表达情况,确定标记滞留细胞(The label-retaining cells,LRCs)。通过免疫组化双染检测LRCs表面CD34和CD146的表达情况。结果 腺上皮和间质的BrdU初始最大标记率分别为(69.89±0.78)%和(73.27±1.06)%,于第6周和8周时分别有约(2.07±0.19)%和(4.63±0.72)%的腺上皮和间质细胞仍保留标记,即为上皮和间质标记滞留细胞,多位于异位内膜与所种植的裸鼠组织的交界处。LRCs并不表达CD34,少数间质LRCs表达CD146。结论 异位子宫内膜中有上皮和间质两种干细胞,主要定位于异位内膜与所种植的组织的交界处,CD34不是异位内膜干细胞的标志物,CD146表达于异位内膜干细胞,但并非其特异性标志物。  相似文献   

18.
张著学  李志义  龚灵  张薇珊  成元华 《贵州医药》2008,32(2):109-112,F0002
目的探讨消化道和腹腔软组织平滑肌肿瘤临床病理和免疫组化特点。方法对17例消化道及腹腔软组织平滑肌肿瘤进行临床病理、免疫组化观察,并对7例患者进行随访。结果男性7例,女性10例;年龄23~63岁,平均49岁。肿瘤位于胃肠道8例(食道2例,胃1例,结肠4例,直肠1例),位于腹腔9例(肠系膜2例,网膜2例,腹膜后5例)。临床表现无特异性,光镜下具有一定特征。随访7例,3例平滑肌瘤均显示预后良好,4例平滑肌肉瘤中3例复发,1例死亡。SMA阳性率100%(17/17),Desmin阳性率94%(16/17),CD117、CD34、S-100蛋白、PDGFRA及PKCtheta均为阴性表达。结论消化道及腹腔软组织平滑肌肿瘤少见,发生于消化道者多为良性,发生于腹腔者多为恶性。该肿瘤强表达SMA及Desmin,PDGFRA或PKC theta可作为与胃肠道间质瘤鉴别诊断的标记物。  相似文献   

19.
Purpose. To identify the growth conditions that would favor thedevelopment of a functional primary culture of pigmented rabbit cornealepithelial cells on a permeable support comparable to the intact tissuein bioelectric properties. Methods. Rabbit corneal epithelial cells were isolated and cultured onprecoated fibronectin/collagen/laminin permeable filters. Cells weregrown at an air-interface with supplemented DMEM/F12 medium.Immunofluorescence and electron microscopy techniques, respectively,were used to confirm cornea-specific marker and morphologicalfeatures. Permeability of the cell layers to model polar compounds wasevaluated using 14C-mannitol, fluorescein isothiocyanate (FITC) andfluorescein isothiocyanate-dextran of 4,000 molecular weight (FD4). Results. We found that culturing the epithelial cells at anair-interface (AIC) was a critical factor in the formation of tight cell layer and thatomitting fetal bovine serum and keeping the concentration of epidermalgrowth factor at 1 ng/ml were equally important. Phenotypically, theAIC cell layers were found to express cornea-specific 64 kD keratin.Compared with cells cultured under the liquid-covered (LCC)condition, those cultured under AIC exhibited a significantly higher peaktransepithelial electrical resistance (TEER) of up to 5 kV.cm2, a higherpotential difference (PD) of up to 26 mV, and an estimated short-circuitcurrent (Ieq) of 5 A/cm2 after 7=n8 days of culture. These values werecomparable to those in the excised cornea. Consistent with the TEER,the AIC cell layers were 4–40 times less permeable to paracellularmarkers than their LCC counterpart. Conclusions. The AIC model merits further characterization of drugtransport mechanisms as well as drug, formulation, physiological, andpathological factors influencing corneal epithelial drug transport.  相似文献   

20.
The term lymphoepithelioma-like carcinoma identifies a group of nasopharingeal epithelial tumors characterized by aggregates of malignant undifferentiated cells surrounded by a dense reactive lymphoplasmacellular infiltrate. Primary cutaneous localization is rare, with approximately 30 cases reported in literature. We describe a case of primary lymphoepithelioma-like carcinoma of the skin in a 92-year-old woman. Immunohistochemical examination was positive for cytokeratine (KL1 and EMA) as regards epithelial cells, while the lymphocitic infiltrate was positive for LCA and CD3. In situ hybridization for Epstein Barr virus in tumor cells was negative. Electron microscopy showed rounded and occasionally spindle-shaped poorly-differentiated squamous epithelial cells, and a lymphoid stroma consisting mostly of normal-appearing small lymphocytes. Examination of the nasopharynx did not show any tumoral mass and after a 7 years follow-up the patient is free of local and distant recurrences. This tumor affects people aged over 50 years and is localized to the face, but scalp, shoulder and forearm may be involved. Research of Epstein-Barr virus is always negative in this tumor, unlike nasopharingeal epithelial carcinoma. The differential diagnosis of lymphoepithelioma-like carcinoma of the skin may present some difficulties and includes squamous cell carcinoma. Lymphoepithelioma-like carcinoma of the skin is a malignant neoplasm which tends to relapse locally and has a moderate tendency to metastatize.  相似文献   

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