网状纤维和肥大细胞双重染色

同一张组织切片上有必要探讨两种或两种以上的组织成分或细胞的相互关系时,可以采用双重染色方法.网状纤维是构成组织的网状支架,广泛分布于实质性脏器中,其中最丰富的区域是肝、脾、淋巴组织、骨髓、基底膜和血管等处.网状纤维染色方法是一种组化方法,利用此方法可以显示和判定病变组织支架的破坏情况及纤维组织的增生情况[1,2].肥大细胞来源于未分化的间充质细胞,在正常的情况下多见于血管周围、黏膜下、疏松结缔组织及肠系膜的小血管周围.其形态特点是细胞较大,直径20～30 μm,呈圆或椭圆形,胞核较小为圆形,胞质内有许多圆形嗜碱性颗粒.这些颗粒中含有的高分子多硫酸酯和硫酸粘多糖类对甲苯胺蓝染料有异染性.肝坏死后产生的纤维组织中发现较多肥大细胞.网状纤维染色和肥大细胞双重染色方法可以阐明网状纤维和肥大细胞之间的相互位置关系.     

子宫Müllerian腺肉瘤伴间质横纹肌样瘤分化1例并文献复习

目的探讨子宫Muellerian腺肉瘤伴间质横纹肌样瘤分化的临床病理特征。方法用光镜、组织化学及免疫组化方法观察其病理组织学表现。结果肿瘤由良性上皮成分和肉瘤性间质成分组成，肉瘤成分过度生长。肿瘤细胞弥漫浸润性分布，细胞大，胞质丰富嗜酸性并可见嗜伊红包涵体。免疫表型：vimentin、CK、NF、CD57、CD99、CgA、Syn阳性，SMA散在阳性，而desmin、EMA、CD10、GFAP、MyoD1、Inhibin—α、HMB45和S-100蛋白阴性。组织化学染色PAS阴性，网状纤维染色显示网状纤维包绕单个或小巢肉瘤性间质细胞。结论子宫腺肉瘤伴间质横纹肌样瘤分化是一种罕见的混合性Muellerian肿瘤，应与子宫内膜间质肉瘤、子宫横纹肌肉瘤和低分化癌等鉴别。     

免疫组织化学阳性标记结果的观察和判断

免疫组织化学标记结果的判断是以观察组织片阳性或阴性染色为前提，就阳性染色而言，应着重观察阳性染色颗粒在细胞中存在的位置和分布形式，因为它直接反映抗原在细胞中定位和分布情况，而抗原的存在与否及其在细胞中的定位正是免疫组化诊断的重要依据。在免疫组化标记中，观察阳性染色颗粒定位有助于区别假阳性反应；决定抗原定位     

介绍一种网状纤维染色改良法

网状纤维染色常被应用于鉴别癌与肉瘤、识别坏死组织的结构和类型、判断肝脏网状支架的塌陷破坏或增生情况等.目前传统网状纤维染色法,染色对比欠清晰,染液不易长期保存~([1]).为此笔者在实践中对氨银液配制及具体染色方法进行了一定改进,不仅可以清晰地显示网状纤维的结构,而且稳定、可靠,此液保存时间长,现介绍如下.     

Ⅰ抗孵育时间对免疫组织化学实验结果的影响

众所周知，抗原与抗体之间的结合具有高度的特异性，免疫组织化学技术应用这一原理，对组织切片或细胞标本中的某些化学成分进行原位的定性、定位或定量研究。在神经科学领域，此项技术是不可缺少的。免疫组织化学染色方法有多种类型，但在染色过程中都需要采取一些基本的技术方法，以增强抗原、抗体的特异性反应，降低或消除非特异性反应，     

在Atelocollagen胶原支架上体外三维培养乳大鼠心肌

目的 在Atelocollagen胶原支架上三维培养具有收缩功能的乳鼠心肌细胞块,为进一步开展心肌组织工程奠定基础。 方法 乳鼠心肌消化培养、纯化后,种植到Atelocollagen胶原支架上进行三维培养。应用倒置显微镜动态观察心肌细胞在Atelocollagen胶原支架上的生长情况。对三维培养不同时间的心肌细胞块进行石蜡包埋、切片,分别进行HE染色、肌球蛋白免疫组织化学染色、肌球蛋白免疫荧光染色、透射电镜观察,对三维培养的细胞进行全面鉴定。 结果 心肌细胞种植到Atelocollagen胶原支架上6h开始贴支架生长,培养2d心肌细胞相互融合,并与Atelocollagen胶原支架形成复合体,连同支架一起搏动。培养6d细胞与细胞间结合较前更紧密。培养10d细胞在Atelocollagen胶原支架孔中交织成网状,18～20d细胞充满大部分支架网孔,进一步形成紧密的心肌细胞块。在整个培养过程中,心肌细胞始终保持良好的自律性收缩。形态学鉴定证明,支架网孔中生长的主要是心肌细胞,极少数为纤维样细胞。 结论 利用Atelocollagen胶原支架能够培养出较为理想的、具有收缩功能的三维心肌细胞块。     

中枢神经系统非典型畸胎样/横纹肌样瘤临床病理及免疫表型特征

目的 探讨中枢神经系统非典型畸胎样／横纹肌样瘤的临床病理特征、诊断及鉴别诊断。方法 对2例非典型畸胎样／横纹肌样瘤应用光镜行HE、网状纤维染色及免疫组织化学染色观察,并结合文献复习。结果 非典型畸胎样／横纹肌样瘤具有特征性的横纹肌样细胞,伴有不同程度的原始神经外胚叶、上皮和间质分化。肿瘤组织富于网状纤维,免疫组织化学标记示波形蛋白、CD99、上皮细胞膜抗原、细胞角蛋白、胶质纤维酸性蛋白、S-100蛋白、神经微丝蛋白、结蛋白、平滑肌肌动蛋白阳性,突触素、肌调节蛋白、胎盘碱性磷酸酶和HMB45阴性。结论 非典型畸胎样／横纹肌样瘤是中枢神经系统一种罕见的高度恶性肿瘤,好发于儿童,偶见于成人,呈异源性组织学和免疫组织化学表型。其诊断需与脑内其他多形性肿瘤鉴别。     

肾上腺皮质嗜酸细胞性肿瘤9例临床病理学特征

目的探讨肾上腺皮质嗜酸细胞性肿瘤的临床病理特征及免疫表型。方法收集9例肾上腺皮质嗜酸细胞性肿瘤,肿瘤组织均进行HE、免疫组化及网状纤维染色,分析其临床病理特征及免疫表型,并复习相关文献。结果 9例患者中男性5例,女性4例,年龄26～64岁,平均46岁,发病部位左侧8例,右侧1例。9例均为含有 90%嗜酸性细胞的单纯性肾上腺皮质嗜酸细胞性肿瘤,参照Lin-Weiss-Bisceglia(LWB)评分系统,2例为恶性,2例为恶性潜能未定,5例为良性。9例肿瘤细胞均表达SF-1,Ki-67增殖指数在良性肿瘤中均≤5%,而在恶性肿瘤中均 10%。网状纤维染色显示良性肿瘤细胞巢网状纤维支架存在,恶性潜能未定及恶性肿瘤网状纤维支架不同程度被破坏。9例中1例失访,余8例随访4～104个月,均未发生转移或复发。结论嗜酸细胞性肿瘤诊断主要依靠组织病理学及免疫组化检测,治疗以手术切除为主。     

免疫组织化学染色常见问题及解决方法

免疫组织化学将传统形态学与传统免疫学结合起来,既可以在光镜下或电镜下观察组织或细胞的形态,实现组织或细胞定位;同时又发挥了免疫学中对特异抗原抗体定性定量的作用.     

婴幼儿促纤维增生性星形细胞瘤的临床病理特点

目的；分析婴幼儿促纤维增生性星形细胞瘤（desmoplastic infantile astrocytoma,DIA)的病理形态特征及其与其他肿瘤的鉴别诊断。方法：对2例DIA进行组织形态学、组织化学和免疫组织化学分析，结合文献对其临床表现、病理形态特点及鉴别诊断进行探讨。结果：本组2例DIA的CT检查表现均为左颞叶巨大的囊性病变；组织学表现在丰富的纤维性间质中，含有灶状或巢状的向星形细胞分化的神经上皮成分；免疫表型显示瘤细胞呈GFAP阳性，Vim阳性，NSE阴性；网织纤维染色示在致密的嗜银纤维区之间可见岛屿状的空染区。结论：DIA是一种罕见的发生在婴幼儿的中枢神经肿瘤。根据其组织学特点，结合组织化学和免疫组化染色结果，可以做出明确的病理诊断。     

Pain and Effusion and Quadriceps Activation and Strength

Context:

Quadriceps dysfunction is a common consequence of knee joint injury and disease, yet its causes remain elusive.

Objective:

To determine the effects of pain on quadriceps strength and activation and to learn if simultaneous pain and knee joint effusion affect the magnitude of quadriceps dysfunction.

Design:

Crossover study.

Setting:

University research laboratory.

Patients or Other Participants:

Fourteen (8 men, 6 women; age = 23.6 ± 4.8 years, height = 170.3 ± 9.16 cm, mass = 72.9 ± 11.84 kg) healthy volunteers.

Intervention(s):

All participants were tested under 4 randomized conditions: normal knee, effused knee, painful knee, and effused and painful knee.

Main Outcome Measure(s):

Quadriceps strength (Nm/kg) and activation (central activation ratio) were assessed after each condition was induced.

Results:

Quadriceps strength and activation were highest under the normal knee condition and differed from the 3 experimental knee conditions (P < .05). No differences were noted among the 3 experimental knee conditions for either variable (P > .05).

Conclusions:

Both pain and effusion led to quadriceps dysfunction, but the interaction of the 2 stimuli did not increase the magnitude of the strength or activation deficits. Therefore, pain and effusion can be considered equally potent in eliciting quadriceps inhibition. Given that pain and effusion accompany numerous knee conditions, the prevalence of quadriceps dysfunction is likely high.Key Words: arthrogenic muscle inhibition, central activation failure, voluntary activation, muscles

Key Points

• Knee pain and effusion resulted in arthrogenic muscle inhibition and weakness of the quadriceps.
• The simultaneous presence of pain and effusion did not increase the magnitude of quadriceps dysfunction.
• To reduce arthrogenic muscle inhibition and improve muscle strength, clinicians should employ interventions that target removing both pain and effusion.

Quadriceps weakness is a common consequence of traumatic knee joint injury^1,2 and chronic degenerative knee joint conditions.^3,4 Arthrogenic muscle inhibition (AMI), a neurologic decline in muscle activation, results in quadriceps weakness and hinders rehabilitation by preventing gains in strength.⁵ The inability to reverse AMI and restore muscle function can lead to decreased physical abilities,⁶ biomechanical deficits,⁷ and possibly reinjury.⁵ Furthermore, researchers^8,9 have suggested that quadriceps weakness resulting from AMI may place patients at risk for developing osteoarthritis in the knee. In light of the substantial influence of quadriceps AMI on these clinically relevant outcomes, we need to improve our understanding of the factors that contribute to this neurologic decline in muscle activity so efforts to target and reverse it can be implemented and gains in strength can be achieved more easily.Joint injury and disease are accompanied by numerous sequelae (ie, pain, swelling, tissue damage, inflammation), so ascertaining which one ultimately leads to neurologic muscle dysfunction is difficult. Whereas a joint effusion can result in AMI,^10–12 the effects of pain are less understood despite many clinicians attributing AMI to pain. Using techniques that introduce knee pain without accompanying injury may provide insights into the role of pain in eliciting AMI.The degree of knee joint damage may play a role in the quantity of AMI that manifests. Hurley et al^13,14 demonstrated that quadriceps AMI, measured using an interpolated-twitch technique, was greater in patients with extensive traumatic knee injury (eg, fractured tibial plateau, ruptured medial collateral ligament, and medial meniscectomy) than patients with isolated joint trauma (ie, isolated anterior cruciate ligament [ACL] rupture). Similarly, patients with more knee joint symptoms (ie, greater number of symptoms and increased severity of symptoms) may present with greater magnitudes of quadriceps inhibition. Recently, investigators¹⁵ have suggested that patients with more pain display less quadriceps strength, supporting this tenet. Given that effusion and pain often present simultaneously with joint injuries and diseases, such as ACL injury and osteoarthritis, examining both the isolated and cumulative effects of these sequelae appears warranted to determine if they influence the magnitude of muscle inhibition.Experimental joint-effusion and pain models are safe and effective experimental methods that allow for the isolated examination of their effects on muscle function. The effusion model, whereby sterile saline is injected directly into the knee joint capsule,⁷ produces a clinically relevant magnitude of the joint effusion that may be present with traumatic injury. Effusion is thought to activate group II afferents responding to stretch or pressure,^16–18 which in turn may facilitate group Ib interneurons and result in quadriceps AMI.⁵ The pain model involves injecting hypertonic saline into the infrapatellar fat pad to produce anteromedial knee pain similar to that described in patients with patellofemoral pain syndrome.¹⁹ Pain is considered to initiate AMI through activation of group III and IV afferents that act as nocioceptors to signal damage or potential damage to joint structures.^16–18 The firing of these afferents then may lead to facilitation of group Ib interneurons, the flexion reflex, or the gamma loop, ultimately resulting in quadriceps inhibition.²⁰ Thus, these models allow us to create symptoms that are associated with knee injury and have the added benefit of providing a way to examine their effects in isolation.Therefore, the purpose of our study was to determine the effects of pain on quadriceps strength and activation and to learn if simultaneous pain and knee joint effusion would affect the magnitude of quadriceps dysfunction. We hypothesized that pain alone would result in quadriceps inhibition and that the magnitude of inhibition would be greater when effusion and pain were present simultaneously.     

即早基因c-fos与脑血管病及学习记忆

即早基因c-fos是广泛存在于原核细胞和真核细胞的高度保守基因.在正常情况下,c-fos基因参与细胞生长、分化、信息传递、学习和记忆等生理过程,而在病理情况下c-fos基因表达及调控变化与多种疾病的发生和发展有关.C-fos在中枢神经系统的某些部位可有基础水平的表达,但表达很低,当受到如脑缺血、脑出血、痫性发作、应激等刺激后,其在数十分钟内做出反应,在对外界刺激-转录耦联的信忠传递过程中起着核内第三信使的重要作用.     

Opportunities and challenges in the prevention and control of cancer and other chronic diseases: children's diet and nutrition and weight and physical activity

OBJECTIVE: The purpose of this article is to review the role of behavioral research in disease prevention and control, with a particular emphasis on lifestyle- and behavior-related cancer and chronic disease risk factors--specifically, relationships among diet and nutrition and weight and physical activity with adult cancer, and tracking developmental origins of these health-promoting and health-compromising behaviors from childhood into adulthood. METHOD: After reviewing the background of the field of cancer prevention and control and establishing plausibility for the role of child health behavior in adult cancer risk, studies selected from the pediatric published literature are reviewed. Articles were retrieved, selected, and summarized to illustrate that results from separate but related fields of study are combinable to yield insights into the prevention and control of cancer and other chronic diseases in adulthood through the conduct of nonintervention and intervention research with children in clinical, public health, and other contexts. RESULTS: As illustrated by the evidence presented in this review, there are numerous reasons (biological, psychological, and social), opportunities (school and community, health care, and family settings), and approaches (nonintervention and intervention) to understand and impact behavior change in children's diet and nutrition and weight and physical activity. CONCLUSIONS: Further development and evaluation of behavioral science intervention protocols conducted with children are necessary to understand the efficacy of these approaches and their public health impact on proximal and distal cancer, cancer-related, and chronic disease outcomes before diffusion. It is clear that more attention should be paid to early life and early developmental phases in cancer prevention.