Effect of stem cell application on Asherman syndrome,an experimental rat model

Purpose

We evaluate the effect of stem cells to induce endometrial proliferation and angiogenesis on Asherman Syndrome (AS).

Methods

The experimental study was performed in stemcell research laboratory. Forty Wistar-Albino rats were divided according to groups. In group1 (n = 10) to establish the model; trichloroacetic acid was injected to right uterine horn. Two weeks later, intrauterine synechia was confirmed. In group2 (n = 10), 2 weeks later, 2 × 106 mesenchymal stem cells (MSC) were injected into right uterine horn followed by three intraperitoneal injections of MSCs. In group3 (n = 10), daily oral estrogen was initiated on the second week. In group4 (n = 10), MSC injections and oral estrogen was given together. The amount of fibrosis, vascularisation, inflammation and immunohistochemical staining with vascular endothelial growth factor (VEGF), proliferating cell nuclear antigen (PCNA) and Ki-67 were evaluated in the uterine tissues.

Results

In all treatment groups; fibrosis decreased but vascularisation and immunhistohemical stainings increased in the experimental side. The amount of fibrosis, vascularisation, Ki-67 and PCNA scores were similar between group2 and 3. In group4, comparing to group2, less fibrosis but more Ki-67, PCNA and VEGF staining was observed.

Conclusion

Stem cells, when added to estrogen, are a highly effective alternative to induce regeneration of endometrium in Asherman Syndrome therapy.     

化疗对舌癌PCNA、bcl-2表达的影响

目的　通过舌癌术前化疗细胞增殖和细胞凋亡相关基因Bcl -2表达的研究 ,探讨化疗作用机制。方法　对舌乳头状瘤、舌癌和舌癌术前化疗后标本共 3 4例 ,采用免疫组织化学染色技术 -酶标链亲和素生物素法(LSAB)染色并进行光镜观察。结果　PCNA在舌癌中的表达最强 ,在舌癌术前化疗后的表达明显减弱 ( p <0 .0 5 )与舌乳头状瘤表达相似 ( p >0 .0 5 ) ,Bcl-2在舌癌化疗后及舌乳头状瘤中低表达 ( p >0 .0 5 ) ,舌癌中表达明显增强( p <0 .0 5 )。结论　化疗药物抑制肿瘤生长的机制可能与抑制肿瘤细胞增殖和诱导肿瘤细胞凋亡有关 ,提示舌癌术前化疗对肿瘤的预后有重要意义     

增殖细胞核抗原在小鼠牙胚后期发育中的表达

目的:探讨增殖细胞核抗原在小鼠牙胚发育后期中的表达及作用。方法:制备出生后BALB/C小鼠牙胚发育各阶段石蜡标本,采用SP免疫组化法观察增殖细胞核抗原在牙冠发育及牙周组织发育过程中的表达。结果:在釉质形成早期,中间层细胞、成釉细胞均呈PCNA阳性表达,且两者在同期表达强弱程度上具有一致性;随着牙冠发育的完成,PCNA的表达减弱;出生后第5天,内外釉上皮融合增殖形成上皮根鞘,牙根开始发育,此时上皮根鞘细胞呈PCNA强阳性表达,牙囊组织亦呈现阳性表达,随着牙周组织的发育和重塑的完成,PCNA在各组织的表达逐渐减弱。结论:细胞增殖是牙齿正常发育所必需的重要调控机制,特别是在釉质形成及牙根形成的过程中,具有重要的作用,提示PCNA可能是组织发生的初始标志物之一。     

涎腺腺样囊性癌中p16蛋白,增殖细胞核抗原(PCNA)染色与病理分型、神经侵犯和复发的关系

目的 :探讨涎腺腺样囊性癌中 p16蛋白与PCNA在染色与病理分型、神经侵犯、复发中的关系。 方法 :利用免疫组化SP法 ,对 5例正常涎腺组织和 3 6例涎腺腺样囊性癌组织p16蛋白及PCNA染色进行检测 ,并对患者进行 10年随访观察。结果 :3 6例腺样囊性癌中 ,不同病理分型癌细胞组织中 p16蛋白阳性染色率间差异有统计学意义 (P <0 .0 5 ) ,PCNA阳性染色率间差异无统计学意义 (P >0 .0 5 ) ,有无神经侵犯癌组织中 p16蛋白阳性染色率间差异无统计学意义 (P >0 .0 5 )。有无复发癌组织中p16蛋白阳性染色率间差异有统计学意义 (P <0 .0 5 )。PCNA阳性染色率间差异有统计学意义 (P <0 .0 5 )。结论 :p16基因突变、缺失与腺样囊性癌病理类型有关 ,其表达水平反映肿瘤组织分化程度。p16蛋白及PCNA表达水平可以作为临床评价肿瘤侵袭性和预测复发的指标     

小鼠釉质发育过程中成釉器中间层细胞增殖、凋亡以及组织非特异性碱性磷酸酶的表达

目的:观察小鼠釉质发育过程中成釉器中间层细胞的增殖、凋亡和其组织非特异性碱性磷酸酶(tissue nonspecific alkaline phosphatase,TNSALP)的表达,探讨中间层在釉质形成过程中的可能作用.方法:取出生后1、3、5、7、9、11、15d的BALB/c小鼠56只,解剖分离含下颌第一磨牙区的下颌骨,脱钙,制片,HE染色进行牙釉质发育情况的组织学观察,分别采用原位末端标记法(TUNEL法)、SP免疫组织化学技术、PV二步法观察中间层细胞的凋亡及检测Bcl-2、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA),并对TNSALP进行组织学定位.通过Image-pro plus 6.0图像分析软件.对图像进行半定量分析.采用SPSS13.0软件包进行t检验、单因素方差分析及SNK-q检验.结果:出生后1d.中间层细胞增殖细胞核抗原由比成釉细胞表达高的强阳性表达逐渐降低;至7d时,其表达为阴性,而且中间层细胞随釉质的发育逐渐凋亡.中间层细胞先于成釉细胞表达TNSALP,出生后5d时即呈现强阳性染色,而后其表达又逐步减弱;成釉细胞7d时开始出现阳性,且呈逐渐增强趋势.结论:中间层细胞的增殖和凋亡及其TNSALP的表达与成釉细胞具有相关性,提示中间层细胞有可能参与成釉细胞的分化及釉质形成.     

增殖细胞核抗原和Bcl-2蛋白在粘液表皮样癌中的表达及其意义

目的：探讨PCNA、Bcl-2蛋白在粘液表皮样癌中的表达及其相互联系。方法：对33例手术切除的粘液表皮样癌组织标本进行免疫组化染色，结果：24例粘液表皮样癌均显示PCNA阳性，低分化组的PCNA表达较高分化组高（P＜0．05）。Bcl-2为弱阳性表达，高分化组的Bcl-2表达较低分化组高，二组之间的Bcl-2阳性表达水平差异显著性（P＜0．05）。XFYW：PCNA与Bcl-2无明显关系。     

Effect of Topical Application of Black Seed Oil on Imiquimod‐Induced Psoriasis‐like Lesions in the Thin Skin of Adult Male Albino Rats

Psoriasis is a chronic inflammatory skin disease that affects about 1%–3% of the world's population. Black seed oil, i.e., the oil extracted from black seeds (Nigella sativa seeds), possesses a broad spectrum of pharmacological actions including anti‐inflammatory, immunostimulatory, and antioxidant properties. This study aimed to investigate the effect of black seed oil on imiquimod (IMQ) induced psoriasis‐like skin lesions. To this end, 30 male albino rats were divided into three groups: group I, control group; group II, psoriasis‐induced group receiving daily topical applications of IMQ cream (5%) on the shaved back skin for 10 consecutive days; and group III, black seed oil group receiving a daily topical dose of black seed oil 5 mg/kg body weight for 10 days after induction of psoriasis. Animals of all groups were sacrificed and specimens obtained from the skin of the central part of the back were processed for histological and immunohistochemical staining with proliferating cell nuclear antigen (PCNA). IMQ application led to epidermal inflammation, hyperplasia and alterations in the normal appearance of keratinocytes with degenerative changes observed at both light and electron microscopic levels. Collagenous fibers were abundant in the dermis and PCNA‐positive cells were detected in all layers of the epidermis. However, topical use of black seed oil strongly inhibited IMQ‐induced psoriasis‐like inflammation and alleviated all epidermal and dermal changes observed after IMQ application, allowing us to conclude that black seed oil can be used as an adjuvant topical therapy for treating psoriasis. Anat Rec, 2017. © 2017 Wiley Periodicals, Inc. Anat Rec, 301:166–174, 2018. © 2017 Wiley Periodicals, Inc.     

Anatomical,histological and immunohistochemical study of testicular development in Columba livia (Aves: Columbiformes)

In this work, testicular ontogeny is analyzed at the anatomical, histological and immunohistochemical levels; the latter through the detection of GnRHR and PCNA in the testicles of embryos, neonates and juveniles of Columba livia. We analyzed 150 embryos, 25 neonates and 5 juveniles by means of observations under a stereoscopic magnifying glass and scanning electron microscope (SEM). The histological analysis was performed using hematoxylin-eosin staining techniques and the PAS reaction. For the immunohistochemical analysis, the expression of GnRHR and PCNA in embryos corresponding to stages 41, 43 and in neonates of 2, 5, 7 and 75 days post-hatch was revealed in testicular histological preparations. That gonadal outline is evident in stage 18. In stage 29, the testes are constituted of a medulla in which the PGCs are surrounded by the Sertoli cells, constituting the seminiferous tubules. From stage 37 a greater organization of the tubules is visualized and at the time of hatching the testicle is constituted of the closed seminiferous tubules, formed of the PGCs and Sertoli cells. The Leydig cells are evident outside the tubules. In the juvenile stages, the differentiation of germline cells and the organization of small vessels that irrigate the developing testicle begin to be visible. In the analyzed stages, the immunodetection of the GnRHR receptor and PCNA revealed specific marking in the plasma membrane and in the perinuclear zone for GnRHR and in the nucleus of the germline cells in juvenile testicles for PCNA. These results can be used as a basis for further study of endocrine regulation events during testicular ontogeny in avian species.