背根神经节内P物质、降钙素基因相关肽免疫阳性神经元与阴茎包皮系带感觉信息的传递

目的:探讨背根神经节(DRG)内P物质(SP)、降钙素基因相关肽(CGRP)免疫阳性神经元与阴茎包皮系带感觉信息传递之间的关系。方法:通过荧光金(FG)逆行标记对大鼠阴茎包皮系带内神经末梢的来源作追踪定位,并结合SP、CGRP免疫荧光标记法,研究大鼠DRG内FG标记阳性神经元中SP、CGRP免疫阳性神经元的形态和分布。结果:FG逆行标记结果发现,大鼠阴茎包皮系带内的神经末梢起源于第6腰髓对应的背根神经节(L6-DRG)和第1骶髓对应的背根神经节(S1-DRG)的神经元。对这些神经元分别作SP、CGRP免疫荧光标记后发现,标记细胞大小不等,分别呈深红色和深绿色,沿神经束成行排列或散在分布。FG/SP、FG/CGRP双标记阳性细胞均为中小型,其数量分别占FG逆行标记阳性细胞总数的1/3和1/2,FG/SP/CGRP三标记阳性细胞占FG逆行标记阳性细胞总数的1/5。结论:大鼠L6-DRG和S1-DRG内的SP、CGRP免疫阳性神经元可能参与阴茎包皮系带感觉信息的传递。     

阴茎包皮及包皮系带内SP免疫阳性神经末梢的分布和来源

目的观察成人阴茎包皮和包皮系带内P物质(substanceP,SP)免疫阳性神经末梢的分布和来源。方法免疫组织化学法观察SP免疫阳性神经末梢的分布,荧光金(fluoro-gold,FG)逆行追踪和SP免疫荧光标记相结合法研究大鼠包皮系带内SP免疫阳性神经末梢的来源。结果成人阴茎包皮及包皮系带内均有密集的SP免疫阳性神经末梢存在,这些神经末梢主要位于表皮基底层,呈树枝状或念珠状分布,大多成束走行。阴茎系带处SP免疫阳性神经末梢的分布密度明显大于阴茎包皮处。FG逆标阳性细胞位于大鼠第六腰髓对应的背根神经节(L6-DRG)和第一骶髓对应的背根神经节(S1-DRG)。阳性细胞大中小不等,大多沿神经束成行排列或散在分布。SP免疫荧光标记细胞大多为中小型,呈深红色。FG/SP双标阳性细胞均为中小型,其数量占FG逆标阳性细胞总数的三分之一。结论SP参与了阴茎包皮及包皮系带感觉信息的传递。大鼠阴茎包皮系带内SP免疫阳性神经末梢源自于L6-DRG和S1-DRG。     

大鼠阴茎包皮和包皮系带内nNOS免疫阳性神经末梢的分布与起源

目的观察成年SD大鼠阴茎包皮和包皮系带内神经型一氧化氮合酶(nNOS)免疫阳性神经末梢的分布和起源。方法免疫组织化学法观察nNOS免疫阳性神经末梢的分布,荧光金(Fluoro-gold,FG)逆行追踪和nNOS免疫荧光标记相结合法研究大鼠包皮系带内nNOS免疫阳性神经末梢的起源。结果成年SD大鼠阴茎包皮和包皮系带内均有nNOS免疫阳性神经末梢存在,这些神经末梢主要位于表皮基底层和真皮乳头层,呈树枝状或念珠状分布。阴茎系带处nNOS免疫阳性神经末梢的图像光密度值(3.4±0.38)明显大于阴茎包皮处(2.2±0.45)。FG逆标阳性细胞位于大鼠第六腰髓对应的背根神经节(L6-DRG)(9.6±1.2)个/mm2和第一骶髓对应的背根神经节(S1-DRG)(1.2±0.2个/mm2)内。阳性细胞大中小不等,大多沿神经束成行排列或散在分布。nNOS免疫荧光标记细胞在L6-DRG和S1-DRG内分别为(24.2±2.6)个/mm2和(24.1±2.1)个/mm2,细胞大多呈中小型。FG/nNOS双标阳性细胞均为中小型,其数量接近FG逆标阳性细胞总数的一半。结论大鼠阴茎包皮系带内含有浓密的nNOS免疫阳性神经末梢,这些神经末梢源自于L6-DRG和S1-DRG。     

大鼠盆神经节NOS和VIP阳性神经元对阴茎海绵体的支配

目的：研究大鼠盆神经节(MPG)中一氧化氮合酶(NOS)、血管活性肠肽(VIP)阳性神经元对阴茎海绵体的支配。方法：应用辣根过氧化物酶(HRP)逆行示踪技术结合免疫组织化学和酶组织化学双标技术,观察大鼠MPD中还原型辅酶Ⅱ(NADPFI,NOS标志物)阳性和VIP阳性神经元对阴茎海绵体的支配。结果：HRP阳性标记神经元主要分布于MPG内靠近阴茎神经的区域。HRP阳性标记神经元中72％(210／292)为VIP免疫阳性,83％(268／323)为NADPH组化反应阳性。结论：大鼠盆神经节中NOS、VIP阳性神经元对阴茎海绵体有直接支配,NO和VIP作为神经递质或调质参与了阴茎勃起。     

人工体神经-内脏神经反射弧传出神经元递质研究

目的研究大鼠人工体神经-内脏神经反射弧传出神经递质性质。方法将建立了人工体神经-内脏神经反射弧的8只模型大鼠随机分为2组(n=4)，1组大鼠的左侧盆神经节(MPG)内注射荧光金(FG)，2组大鼠的尿道外括约肌(EUS)注射辣根过氧化物酶(HRP)，通过免疫组织化学方法显示FG、HRP阳性标记细胞中的中的胆碱已酰转移酶((3hAT)。结果FG、HRP逆行追踪结果显示，FG、HRP阳性标记细胞主要分布于脊髓L3尾部至L5头侧左侧前角。FG标记和ChAT阳性双标细胞约占FG标记阳性细胞的88％(133／151)；HRP标记和ChAT阳性双标细胞约占HRP标记阳性细胞的91％(93／102)。结论人工体神经，内脏神经反射弧传出神经元的神经递质主要是乙酰胆碱，体神经和内脏神经(副交感节前神经)吻合后再生神经主要递质没有改变。     

端侧神经吻合后再生状况的实验研究

目的研究端侧神经吻合后,神经再生的可能性和再生纤维的类型.方法将失神经后的兔耳大神经制成端侧吻合模型,12只动物被分成2、4、6、8和12周5个实验组和1个正常对照组,在受神经侧注入HRP逆行示踪,实验到期后对耳大神经纤维和相应的神经节进行HRP染色和CGRP的免疫组化染色,光镜下观察结果.结果逆行示踪显示,神经端侧吻合2～4周时,供神经侧C2、C3背根神经节内无HRP标记阳性的细胞,至第6周,背根神经节内开始出现HRP阳性细胞,8～12周阳性细胞的数量更多,尤其是胞体直径在50μm以上的阳性细胞数显著增加;2～4周时CGRP免疫组化的染色显示,背根神经节内的CGRP阳性标记细胞逐步增强,6周以后阳性标记的细胞数量增加,阳性细胞的胞体直径在12～40 μm范围内,属于中小直径的细胞胞体,同时观察到供神经纤维内的CGRP阳性纤维逐步增多,并跨越神经吻合口长入受神经体内,8～12周背根神经节内的CGRP阳性标记细胞的增加不显著,吻合口及受神经体内的神经纤维也增加不明显.结论端侧神经吻合后再生纤维可长入移植体,纤维再生的顺序是细小类的纤维首先再生.     

利多卡因注射大鼠背根神经节致神经炎症的研究

背景选择性脊神经阻滞导致的脊神经或背根神经节（DRG）损伤是至今尚未充分研究的一种潜在的严重并发症。本实验假设局部麻醉药注入脊神经、DRG内可能会导致炎性反应与痛觉过敏。方法大鼠部分椎板切除后,于L5脊神经或背根神经节内注射4μl利多卡因或盐水,评价其炎性反应及行为反应。行为学测试在手术前、手术后分别进行,通过观察并记录大鼠对足部伤害性机械刺激的反应,并对痛觉过敏进行评估。摘取背根神经节并染色,对神经元周围起免疫反应的神经胶质细胞环进行计数。结果与脊神经利多卡因注射组相比,DRG利多卡因注射组注射利多卡因4天后出现了同侧足部的痛觉过敏。神经胶质原纤维酸性蛋白免疫阳性的神经胶质细胞环,即激活的卫星细胞的数量,在DRGs内的表达,DRG和脊神经利多卡因注射组均明显增加。同盐水注射组相比,利多卡因注射组神经胶质原纤维酸性蛋白阳性细胞明显增多,并在DRGs内发现了散在的、能代表激活的小神经胶质细胞的OX-42免疫阳性细胞。通过检测能标记激活的T淋巴细胞的Pan-T,并未发现T淋巴细胞。结论DRG内注射利多卡因可导致痛觉过敏,可能是通过激活固有的卫星神经胶质细胞。临床上,选择性脊神经阻滞时,应避免DRG内直接注射局部麻醉药。     

人工体神经-内脏神经反射弧恢复截瘫后直肠功能的神经追踪研究

目的 研究正常和建立人工体神经-内脏神经反射弧脊髓损伤大鼠肛门外括约肌-脊髓、直肠-盆神经节-脊髓之间的神经通路。方法 正常及建立人工体神经-内脏神经反射弧后截瘫的(以下简称模型组)雄性SD大鼠，用荧光金(FG)注射至肛门外括约肌(EAS)、直肠壁内、盆神经节(MPG)，行逆行神经追踪；模型组大鼠，采用麦芽凝集素结合的辣根过氧化物酶(WGA-HRP)作为顺行神经示踪剂，在大鼠脊髓左侧L4前角注入WGA-HRP。结果 正常组和模型组大鼠直肠壁注入FG后，MPG主体部可见大量标记神经元。正常组将FG注射入MPG后。FG标记神经元位于脊髓L6～S1骶髓副交感核(SPN)、后联合灰质核。正常鼠FG注射入肛门外括约肌后．标记神经元主要位于L5～S1脊髓的背内侧核(DM)区．背外侧核(DL)区可见少量标记神经元。模型组大鼠将FG注入盆神经节或肛门外括约肌后，左L4前角均可见FG标记神经元；左L4前角注入WGA-HRP后，肛门外括约肌和左侧MPG中均可见HRP阳性神经末梢。结论 正常大鼠盆神经节支配直肠的神经元主要分布于MPG主体部；脊髓内支配盆神经节的神经元主要位于脊髓L6～S1节段SPN；支配肛门外括约肌的运动神经元主要位于L5～S1脊髓DM区。建立人工反射弧后。大鼠L5～S1脊髓L4前根与L6前根吻合后所形成的“异类神经纤维”在MPG换元后，支配直肠壁和EAS。     

大鼠下尿路、腰骶髓脊髓排尿中枢逆行神经追踪研究

目的应用荧光金（FG）、快蓝（FB）、辣根过氧化物酶（HRP）逆行神经追踪技术研究盆神经节（MPG）、尿道外括约肌（EUS）与骶髓排尿中枢之间的神经通路。方法选用正常雄性SD大鼠．盆神经节、尿道外括约肌、尿道黏膜下、膀胱逼尿肌内注射神经追踪剂组织化学反应及荧光显微镜。结果标记神经元在脊髓L6部至S1侧角（以L6多见）可见FB阳性神经元，L6脊髓前角的腹外侧部，即Onuf＇s核，同时中间外侧柱（IML）、中央管周围出现FG阳性神经元。盆神经节在荧光显微镜下可发现HRP暗黑色阳性神经细胞和呈蓝色的荧光阳性神经细胞；脊髓L6部至S1灰质后联合核、中央管周围、IML区同时出现FG、FB细胞，但未发现FG、FB的双标细胞。结论大鼠脊髓内排尿中枢主要位于脊髓L6-S1节段，主要包括支配盆神经节的PGN和支配EUS的前角神经元。灰质后联合核、中央管周围、IML区内的亦可见少许神经元。盆神经节支配膀胱逼尿肌，是排尿反射的最后一级神经元。     

硬膜外腔植入髓核对大鼠脊髓背根神经节SP和CGRP表达的影响

目的 观察硬膜外腔植入异体髓核对大鼠L6-S1,脊髓背根神经节细胞SP和CGRP表达的影响,以期为椎间盘源性疼痛发病机制提供细胞生物学基础.方法 18只雄性SD大鼠(体重260-280g)随机分为三组:脂肪组、髓核组、假手术组,每组n=6.另外3只雄性SD大鼠用来提供异体脂肪和髓核.术后第30d取L4-L6脊髓背根神经节和脊髓背角,采用免疫组织化学染色方法 观察髓核对脊髓背角和背根神经节细胞肽类神经递质SP和CGRP表达的影响.结果 术后30d脂肪组大鼠与假手术组相比,脊髓背角和背根神经节细胞SP和CGRP表达没有显著性差异(P>0.05);髓核组与脂肪组和假手术组相比脊髓背角和背根神经节细胞SP和CGRP表达显著增加(P<0.05).结论 硬膜外腔植入异体髓核可引起脊髓背根神经节细胞SP和CGRP表达增加.     

2015年乳腺癌辅助化疗进展

【摘要】〓乳腺癌是危害我国女性健康的头号杀手,尽管近年来辅助化疗的研究进展突飞猛进,但临床中仍有不少问题未能明确,如辅助化疗的合适人群、化疗的开始时间、蒽环及紫杉类的地位和用法、强化维持治疗的作用、疗效及预后的生物标志物等。本文结合乳腺癌辅助化疗在临床上的常见问题和2015年各大乳腺癌会议阐述乳腺癌辅助化疗的最新进展。     

Alterations in T-Cell Subset Frequency in Peripheral Blood in Obesity

Background: Obesity affects the regulation of immune and inflammatory responses. This study characterizes differences in peripheral blood lymphocyte phenotype in obese humans. Methods: Frequencies of lymphocyte subsets among peripheral blood mononuclear cells were compared between 10 obese (BMI ≥35) and 10 lean subjects, as determined by antibodies directed against cluster differentiation (CD) markers. Results: Obese patients demonstrated an increased frequency of CD3+CD4+ T-cells (mean difference 12%, P=0.004), a decreased frequency of CD3+CD8+ T-cells (mean difference 9.4%, P=0.016) and an increased frequency of CD3+CD8+CD95+ T-cells (mean difference 13.3%, P=0.032). No other differences among T-cell or monocyte subsets were noted. Conclusions: Obesity is associated with alterations in frequencies of peripheral CD4+ and CD8+ T-cells and aberrations in the expression of CD95 among CD8+ T-cells. These data suggest both CD4+ and CD8+ T-cell compartments, as well as the regulation of CD95 expression on CD8+ T-cells, as targets for further study into obesity's effects on the immune system.     

β-半乳糖苷酶在体内外成骨细胞中的表达

目的 研究β—半乳糖苷酶(β—gal)在成骨细胞中的表达状况，为阐明MorquioB综合征的发病机制提供依据。方法 裸鼠各器官和骨组织标本行X-gal染色检测。抽取羊和人骨髓行骨髓基质细胞(BMSCs)培养，分为4组：I：Adv-hBMP-2转染组；Ⅱ：Adv—β—gal转染组；Ⅲ：未转染组；Ⅳ：地塞米松诱导组。分别行X-gal染色和RT-PCR检测β—gal的表达。结果 裸鼠骺板两侧、骨膜内面及松质骨的成骨细胞和破骨细胞可见多量β—gal的表达。未转染BMSCs组有少量β—gal的表达，其他3组细胞的β—gal表达增高。结论成骨细胞和破骨细胞可表达多量β—gal，该两种细胞的β—gal缺乏可能是MorquioB综合征骨骼异常的直接原因。     

西宁地区烧伤病人动脉血气分析观察

对高海拔地区的27例烧伤病人动脉血气变化进行了分析和观察。结果证明:无论是存活病人还是死亡病人伤后均存在有低氧血症问题。并且在死亡病人和烧伤合并吸入性损伤病人其低氧血症的发生早于单纯烧伤病人。提示:吸入性损伤病人应立即行气管切开术以保障氧气供给,单纯烧伤病人可常规吸氧以维持正常血 PaO_2,ARDS 均发生在合并吸入性损伤的病人,高频喷射通气技术对纠正低氧血症有一定效果。     

Controversies in Fistula in Ano

Managing a complex fistula in ano can be a daunting task for most surgeons; largely due to the two major dreaded complications—recurrence & fecal incontinence. It is important to understand the anatomy of the anal sphincters & the aetiopathological process of the disease to provide better patient care. There are quite a few controversies associated with fistula in ano & its management, which compound the difficulty in treating fistula in ano. This article attempts to clear some of those major controversies.     

Smoking-Attributable mortality in Spain in 2016

IntroductionSmoking-attributable mortality (SAM) is a valuable indicator that can be used to characterize the course and health burden of the smoking epidemic. The aim of this paper was to estimate SAM in Spain in 2016 in the population aged 35 and over, using the best available evidence.MethodsA smoking prevalence-dependent analysis based on the estimation of population-attributable fractions was performed. Smoking prevalence (never, former, and current smokers) was calculated from a combination of the Spanish Health Survey (2016) and the European Health Survey (2014); the relative risk of death among current and former smokers was taken from the follow-up of various cohorts; and mortality rates were obtained from National Center for Statistics data. SAM estimates are presented globally, and by sex, age groups, and major disease categories: cancer, cardiometabolic diseases and respiratory diseases.ResultsIn 2016, 56,124 deaths were attributed to tobacco consumption, 84% in men (47,000), and 50% in the population aged over 74 (27,795). Overall, 50% of SAM was due to cancer (28,281), 65% of which was lung cancer. One in 4 attributable deaths (13,849) occurred before the age of 65.ConclusionsOne in 7 deaths in Spain in 2016 were attributable to smoking. This estimation of SAM clearly highlights the great impact of smoking on mortality in Spain, mainly due to lung cancer and chronic obstructive pulmonary disease.     

Single center experience in single-incision laparoscopic surgery in children in Turkey

Purpose

Minimally invasive surgery has evolved into single-incision laparoscopic surgery (SILS) in the recent years. Few reports have addressed the practicality of SILS in children. Our current experience with regard to feasibility and effectiveness of SILS in children is presented.

Methods

A retrospective review of the operative database for patients operated on using SILS in our department from March 2009 to July 2010 was performed. Data regarding the type of the procedure, age, sex, operative performance, hospital stay, and complications were collected.

Main Results

Among 43 patients, cholecystectomy was performed in 11; appendectomy, in 10; unroofing for ovarian cysts, in 5; unroofing for splenic cysts, in 4; oophorectomy, in 6 (ovarian torsion, 2; teratoma, 4); ovary-preserving teratoma excision, in 1; splenectomy, in 1; gonadectomy, in 3; and varicocelectomy, in 2. There were no conversions to standard laparoscopic or open techniques. The only postoperative complication was a wound infection that occurred after an appendectomy.

Conclusion

Although currently more expensive, SILS can be performed in children in almost every pediatric surgical procedure that can be accomplished with conventional laparoscopic techniques. The most significant contribution of SILS procedure is cosmesis. Postoperative pain and length of hospital stay were not improved.     

MicroRNAs in hepatic pathophysiology in diabetes

MicroRNAs(miRNAs or miRs) are small approximately 22 nucleotide RNA species that are believed to regulate diverse metabolic and physiological processes.In the recent past,several reports have surfaced that demonstrate the role of miRNAs in various biological processes and numerous disease states.For a disease as complex as diabetes,the emergence of miRNAs as key regulators leading to the disease phenotype has added a novel dimension to the area of diabetes research.On the other hand,the liver,a metabolic hub,contributes in a major way towards maintaining normal glucose levels in the body as it can both stimulate and inhibit hepatic glucose output.This equilibrium is frequently disturbed in diabetes and hence,the liver assumes special significance considering the correlation between altered hepatic physiology and diabetes.While the understanding of the mechanisms behind this altered hepatic behavior is not yet completely understood,recent reports on the status and role of miRNAs in the diabetic liver have further added to the complexities of the knowledge of hepatic pathophysiology in diabetes.Here,we bring together the various miRNAs that play a role in the altered hepatic behavior during diabetes.