Capsaicin-sensitive afferentation represents an indifferent defensive pathway from eradication in patients with H.pylori gastritis

AIM:To study the role of capsaicin-sensitive afferent nerves in Helicobacter pylori (H.pylori) positive chronic gastritis before and after eradication.METHODS:Gastric biopsy samples were obtained from corpus and antrum mucosa of 20 healthy human subjects and 18 patients with H.pylori positive chronic gastritis (n=18) before and after eradication.Tradi-tional gastric mucosal histology (and Warthin-Starry silver impregnation) and special histochemical examina-tions were carried out.Immunohistochemistry for cap-saicin receptor (TRVP1),calcitonin gene-related peptide (CGRP) and substance P (SP) were carried out by the labeled polymer immunohistological method (Lab VisionCo.,USA) using polyclonal rabbit and rat monoclonal antibodies (Abcam Ltd.,UK).RESULTS:Eradication treatment was successful in 16 patients (89%).Seven patients (7/18,39%) re-mained with moderate complaints,meanwhile 11 pa-tients (11/28,61%) had no complaints.At histological evaluation,normal gastric mucosa was detected in 4 patients after eradication treatment (4/18,22%),and moderate chronic gastritis could be seen in 14 (14/18,78%) patients.Positive immuno-staining for capsaicin receptor was seen in 35% (7/20) of controls,89% (16/18,P < 0.001) in patients before and 72% (13/18,P < 0.03) after eradication.CGRP was positive in 40% (8/20) of controls,and in 100% (18/18,P < 0.001) of patients before and in 100% (18/18,P < 0.001) after eradication.The immune-staining of gastric mucosa for substance-P was positive in 25% (5/20) of healthy con-trols,and in 5.5% (3/18,P > 0.05) of patients before and in 0% of patients (0/18,P > 0.05) after H.pylori eradication.CONCLUSION:Distibution of TRVP1 and CGRP is altered during the development of H.pylori positive chronic gastritis.The immune-staining for TRVP1,CGRP and SP rwemained unchanged before and after H.py-lori eradication treatment.The capsaicin-sensitive affer-entation is an independent from the eradication treat-ment.The 6 wk time period might not be enough time for the restituion of chronic H.pylori positive chronic gastritis.The H.pylori infection might not represent the main pathological factor in the development of chronic

Axonotmesis-evoked plantar vasodilatation as a novel assessment of C-fiber afferent function after sciatic nerve injury in rats

Quantitative assessment of the recovery of nerve function, especially sensory and autonomic nerve function, remains a challenge in the field of nerve regeneration research. We previously found that neural control of vasomotor activity could be potentially harnessed to evaluate nerve function. In the present study, five different models of left sciatic nerve injury in rats were established: nerve crush injury, nerve transection/ suturing, nerve defect/autografting, nerve defect/conduit repair, and nerve defect/non-regeneration. Laser Doppler perfusion imaging was used to analyze blood perfusion of the hind feet. The toe pinch test and walking track analysis were used to assess sensory and motor functions of the rat hind limb, respectively. Transmission electron microscopy was used to observe the density of unmyelinated axons in the injured sciatic nerve. Our results showed that axonotmesis-evoked vasodilatation in the foot 6 months after nerve injury/repair recovered to normal levels in the nerve crush injury group and partially in the other three repair groups;whereas the nerve defect/non-regeneration group exhibited no recovery in vasodilatation. Furthermore, the recovery index of axonotmesis-evoked vasodilatation was positively correlated with toe pinch reflex scores and the density of unmyelinated nerve fibers in the regenerated nerve. As C-fiber afferents are predominantly responsible for dilatation of the superficial vasculature in the glabrous skin in rats, the present findings indicate that axonotmesis-evoked vasodilatation can be used as a novel way to assess C-afferent function recovery after peripheral nerve injury. This study was approved by the Ethics Committee for Laboratory Animals of Nantong University of China (approval No. 20130410-006) on April 10, 2013.

THE CORRELATION OF CENTRAL AFFERENT OF ACUPOINTS QUANLIAO(SI 18) AND SHIBAI(ST 2): TRANSGANGLIONIC DEGENERATIVE METHOD STUDY

Using transganglionic degenerative methed, we investigated the primary afferentinformation of acupoints of Quanliao (SI 18) and Shibai(ST 2). The results showed that the largeamounts of degenerative fibers could be found bilaterally in trigeminal spinal nucleus and its tract,trigeminal principal sensory nucleus, trigeminal mesencephalic nucleus and some degenerations couldbe observed ipsilaterally in solitary tract and its nucleus, medullary lateral reticular nucleus, giganto-cellular reticular nucleus and nucleus raphe magnus following transection of infraorbital nerve. A verymoderate degenerative fibers could be identifed in ipsilateral accessory spinal nucleus and spinal reticu-lar nucleus. A few degenerative fibers were seen on the contralateral magnocellular part of caudatepart of trigeminal spinal nucleus by passing through the posterior commissure of canalis centralis.These results suggest that the primary information of acupoints of Quanliao and Shibai could be trans-mittered into the above

Responses of Primary Afferent Fibers to Acupuncture-Like Peripheral Stimulation at Different Frequencies:Characterization by Single-Unit Recording in Rats

The pain-relieving effect of acupuncture is known to involve primary afferent nerves(PANs) via their roles in signal transmission to the CNS.Using single-unit recording in rats,we characterized the generation and transmission of electrical signals in Aβ and Aδ fibers induced by acupuncture-like stimuli.Acupuncture-like signals were elicited in PANs using three techniques:manual acupuncture(MAc),emulated acupuncture(EAc),and electro-acupuncture(EA)-like peripheral electrical stimulation(PES).The discharges evoked by MAc and EAc were mostly in a burst pattern with average intra-burst and inter-burst firing rates of 90 Hz and 2 Hz,respectively.The frequency of discharges in PANs was correlated with the frequency of PES.The highest discharge frequency was 246 Hz in Aβ fibers and 180 Hz in Aδ fibers.Therefore,EA in a dense-disperse mode(at alternating frequency between 2 Hz and 15 Hz or between 2 Hz and 100 Hz) best mimics MAc.Frequencies of EA output>250 Hz appear to be obsolete for pain relief.

EXPERIMENTAL WORK AND RESEARCHStudy on Massage-Evoked Potentiogram from Somatic Nervesand Analysis on the Involvement of Its Afferent Fibers

The experiments were applied on 66 rabbits. While massage was applied on six acu-points, such as Heyang (UB 55) , Xiguan (Liv 7) , and others on legs with the manipulations of finger pres-sure and finger kneading, the massage-evoked potentiogram on the nerve cutaneus posterior femoralisand the nerve gastrocnemius medialis were obtained. The massageevoked potentiogram synchronizedwith massage evoked impulses and varied with different manipulations. All four groups of afferent fiberscontained in nerve cutaneous posterior femoralis and nerve gastrocnemius medialis were involved in con-ducting the impulses caused by the two manipulations. These results indicate that the massage manipula-tions have regular influence on the activities of both the sensitive tissues in acupoints and the somatic af-ferent nerves. Therefore, the results will scientifically enrich the application of massotherapy as far asmanipulations are concerned. Furthermore, the results suggest vast vistas for the systematical study otthe mechanism of massage trom the neurobiological view-point.

催产素与孤独症谱系障碍的发病关系及推拿干预效应的理论探索

孤独症谱系障碍(autism spectrum disorder,ASD),是以刻板重复行为、社交障碍、兴趣局限等为主要临床表现的神经发育障碍性疾病。推拿作为儿科常用的治疗手段之一,可显著改善ASD患儿的异常行为,改善其社会能力。在基础研究方面,催产素(oxytocin,OXT)含量下降是ASD发病的重要假说之一,而推拿的疗效可能与机械刺激激活C类触觉传入纤维从而上调OXT含量有关。从OXT角度讨论ASD发病的神经生物学机制,阐明OXT与ASD发病的潜在联系,即OXT异常导致ASD患者边缘奖赏系统功能受损进而诱发社交障碍,并论述推拿可能通过调节OXT含量产生治疗效应的具体机制。

传入 C 纤维的兴奋在电针“足三里” 激活中缝大核中的作用

为了分析电针“足三里”是否以其伤害性刺激性质引起镇痛作用,我们试探了电刺激“足三里”穴区所引起的传人(顺行)冲动是否可以减小刺激腓总神经引起的逆行 C 波。碰撞实验表明刺激“足三里”确可兴奋了腓神经的一些 C 纤维。此外我们还观察到刺激“足三里”的强度达到或超过 C 纤维阈值时。可明显激活中缝大核神经元,当刺激强度达到可引起最大 c 波时,激活 NRM 的效应也达到最大。上述结果提示电针“足三里”除可以其非伤害性刺激性质引起镇痛作用外,可能主要是以其伤害性刺激,经 C 类纤维激活 NRM,再经过痛负反馈调制引起镇痛。

不同频率捻转手法对大鼠胃运动和胃迷走神经传入纤维放电影响的分析研究

目的:通过分析不同频率捻转手法针刺足三里穴对大鼠胃运动和胃迷走神经传入纤维放电频率的影响,探讨针刺手法作用规律。方法:选用健康成年雄性SD大鼠,分为60次/min针刺组和120次/min针刺组。20%乌拉坦麻醉后暴露胃体部,经十二指肠放置球囊于胃窦部,分别用60、120次/min两种频率的均匀捻转手法针刺足三里,使用Biopac生理记录仪记录针前、行针时、留针时和出针后的胃运动和胃迷走神经传入纤维放电信号,对胃运动频率、波幅和迷走神经放电频率进行提取分析。结果:针刺正常大鼠足三里后,60次/min捻转手法对正常大鼠胃运动波幅、频率呈现抑制作用,120次/min捻转手法对大鼠胃运动波幅呈现促进趋势;行针及出针5 min、15 min时,捻转60次/min组胃迷走神经放电频率减少,针刺组间比较具有统计学差异(P<0.05)。结论:不同频率捻转针刺手法对健康大鼠胃运动和胃迷走神经传入纤维放电频率作用效果有差异。

迷走神经及其传入纤维对大鼠胃肌电活动和胃排空的影响

目的 观察迷走神经及其传入纤维对清醒大鼠胃肌电活动和胃排空的影响 ,研究胃肌电活动和胃排空的神经调节机制。方法 雄性Wistar大鼠 30只 ,随机分为对照组、迷走神经干切断组 (vagotomy组 )和迷走神经传入纤维阻断组 (capsaicin组 ) ,3组大鼠均于胃浆膜面植入电极 1对 ,于清醒状态下记录胃肌电活动 ,随后采用酚红排空方法检测大鼠胃排空。结果 ①vagotomy组大鼠胃肌电活动的主频显著低于对照组和capsaicin组 ;②capsaicin组大鼠胃肌电活动的主频与对照组间差异无显著性意义 ;③vagotomy组大鼠胃排空显著低于对照组 ,而capsaicin组大鼠胃排空显著高于对照组。结论 迷走神经对胃肌电活动的影响是由其传出纤维介导的。迷走神经切断术和辣椒素对大鼠胃排空的不同影响显示迷走神经对胃运动的调控存在多重作用 ,可能由不同神经通路介导。

耳蜗毛细胞和螺旋神经节及其神经纤维的联合定量观察

目的：将耳蜗基底膜取材技术与耳蜗切片技术相结合以观察同一耳蜗中几个主要结构在病理发展中的相互关系。方法：耳蜗铺片用于定量观察毛细胞，骨性螺旋板切片用于定量观察缰孔内的神经纤维，中轴切片用于定量观察螺旋神经节。结果：将卡铂耳中毒灰鼠的耳蜗标本与正常灰鼠进行比较，显示内毛细胞，神经纤维和螺旋神经节的损失百分比基本一致。结论：卡铂对内毛细胞，神经纤维和螺旋神经节都有破坏作用，至于在卡铂病变早期，究竟哪一部分对损伤更敏感，有待进一步研究。

辣椒素处理对电针“四白”穴诱导三叉神经脊束核尾侧亚核c-fos表达的影响

目的:探讨四白穴针刺效应的传入途径。方法:SD大鼠36只,随机分成空白组、假手术组、辣椒素组、溶媒组、电针"四白"穴+辣椒素组、电针"四白"穴+溶媒组。游离一侧眶下神经,辣椒素组将浸泡有1.5%辣椒素的棉条包绕在眶下神经上。溶媒组用浸泡在溶媒中的棉条包绕在眶下神经上。采用免疫组化技术观察三叉神经脊束核尾侧亚核(cSTN)的c-fos表达。结果:空白组在cSTN各层可见到零星、散在的阳性细胞。溶媒组、辣椒素组以及假手术组cSTN的c-fos阳性细胞数与空白组相似。电针"四白"穴+溶媒组c-fos在cSTN内主要集中在浅层(Ⅰ-Ⅱ层),其阳性细胞的数量明显多于其余各组(P<0.01);cSTN的深层(Ⅲ-Ⅳ层)c-fos数量多于空白组、溶媒组和假手术组(P<0.01,P<0.05,P<0.05)。电针"四白"穴+辣椒素组cSTN的Ⅰ-Ⅱ板层的c-fos阳性细胞数与电针"四白"穴+溶媒组相比明显减少(P<0.01),但cSTN的Ⅲ-Ⅳ板层的c-fos阳性细胞数与电针"四白"穴+溶媒组相比无明显变化(P>0.05)。结论:C纤维可能是四白穴针刺效应(调节胃功能活动)的主要传入途径。

舌快反应传入神经终末与三叉神经脊束核的突触联系

目的 :研究舌感觉快反应传入纤维终止于三叉神经脊束核形成的突触类型及各类型突触出现的频率 ,为神经生理学和神经解剖学研究提供参考资料。方法 :采用细胞内注射 ,电镜连续切片技术。结果 :猫舌感觉传入快反应纤维投射到三叉神经脊束核形成的突触类型分单纯型、中间型和复杂型 3种 ,以中间型突触出现率最多。形成突触的各组成部分以轴 -树及轴 -轴突触形式为主 ,未见轴 -体突触。结论 :中间型突触和轴 -树及轴 -轴突触是三叉神经脊束核处理感觉信息的形态学基础。

逆向电刺激大鼠背侧皮神经对相邻脊髓节段Aδ类传入纤维机械感受特性的影响

目的 :分析循经分布的Aδ类初级传入纤维在跨脊髓节段信息传递中机械感受特性的改变和意义 ,从而探讨外周神经参与经络活动的机制。方法 :自近中端断离循足太阳膀胱经分布的胸 9至胸 1 2脊神经背侧皮支 ,在其中一支上施加逆向电刺激 ,以相邻脊髓节段的一支上Aδ类单位放电为指征 ,测定单位的适应特性、机械感受阈值、感受野形状和面积、感受野的敏感点数及其位置、传导速度等机械感受特性。在相邻脊髓节段皮神经上施加逆向电刺激后 ,观察上述各种机械感受特性的改变。结果 :在 6 7只大鼠上观察了 90个Aδ类初级传入纤维的机械感受特性和跨节段刺激对这些特性的影响。快适应和慢适应单位分别占 3 1 1 %和 6 8 9% ,适应时程为 2 0 82±2 51sec (n =90 )。机械感受阈值 0 2 1± 0 0 3mN。平均感受野面积 1 7 76± 3 40mm2 ,感受野内敏感点数为 1 79± 0 1 1个 ,77 3 % (n =88)的敏感点循足太阳膀胱经背部二直行脉之间分布。逆向电刺激相邻脊髓节段皮神经背侧皮支后 ( 0 45mA ,0 1ms,2 0Hz,持续 1 0sec) ,71 9% (n =6 4)的单位感受野增大 ,全部单位感受野均值增大为 71 98± 1 3 0 7mm2 ,显著大于对照 ,P <0 0 0 1。每个感受野内平均敏感点数显著增至 3 0 8± 0 2 5个 ,P <0 0 1。 71 7%

吗啡依赖大鼠脊髓Ⅱ板层初级传入纤维突触性终末数量变化的电镜定量研究

目的 应用电镜定量方法探讨在吗啡依赖条件下 ,大鼠背根节感觉神经元发出的初级传入纤维能否在脊髓 板层侧支出芽和建立新的突触。 方法 用光镜体视学方法测知吗啡依赖组和对照组大鼠 L4脊髓 板层横切面积没有差异的情况下 ,直接比较了两组动物脊髓 板层神经毡单位面积内突触性终末的数量变化。 结果 吗啡依赖组脊髓 板层来自初级传入纤维的突触性终末数 ,明显多于对照组的初级传入纤维突触性终末数。其中来自无髓传入纤维的突触性终末数与对照组同类型的突触性终末数比较增加 6 1%。来自有髓传入纤维的突触性终末数比对照组增多 70 %。尤其是来自含有致密核芯小泡的初级传入纤维突触性终末数比对照组多 98%。 结论 吗啡可能促进初级传入纤维在脊髓

电刺激大鼠腓肠神经引起A_δ、C类传入神经纤维的背根反射

在距脊髓约 15mm处切断大鼠L5背根 ,将中枢端分成 4～ 5条细束 ,电刺激腓肠神经在背根细束上记录背根反射 (dorsalrootreflex ,DRR)。共记录到DRR 5 1例 ,根据引起DRR所兴奋的腓肠神经纤维类别和DRR在背根逆向传出的纤维类别将DRR分为 5类 :Aαβ Aαβ·DRR、Aβδ Aδ·DRR、Aβδ C·DRR、Aαβδδ C·DRR和C C·DRR。结果证明 ,电刺激外周神经激活各类纤维不但能引起A类 (包括Aδ)纤维的DRR ,而且也能引起C类纤维的DRR。记录的Aδ·DRR和C·DRR为细纤维传入终末产生突触前抑制提供了客观指标 ,为DRR逆向传出冲动到达外周组织 ,释放神经肽类递质 。

大鼠脊神经节细胞周围突的分支及其向躯体与内脏的分支投射

本文用大鼠做两组实验。一组用CB-HRP注入其膀胱壁内,TMB法成色,追踪其脊神经节内的标记细胞。发现有的节细胞的周围突在节内出现分支,分二支或多支。另一组实验用荧光素双标记法,分别注射核黄和真蓝于大鼠的坐骨神经干和膀胱壁内,追踪其脊神经节内的标记细胞。结果在L_4—L_6节发现双标细胞,以L_5节最多见,双标细胞出现的比例,均占坐骨神经标记细胞总数的3％左右。结果表明少量脊神经节细胞的周围突有向躯体与内脏的分支投射。

电刺激大鼠皮神经外周端对相邻脊髓节段皮神经内Aβ纤维的机械感受特性的影响

为观察Aβ类初级传入纤维是否参与相邻脊髓节段外周末梢之间的信息传递及其相关机制 ,实验自近中端切断一侧T8～T12 脊髓节段背侧皮神经 ,将一支被切断的皮神经的外周端分离成数支细束 ,以单个Aβ纤维放电为指征 ,检测单位的传导速度、适应特性、机械感受阈值、感受野的形状和面积 ;在相邻脊髓节段、也与中枢断离的皮神经干上施加逆向电刺激 ( 0 45mA ,0 1ms,2 0Hz,10s) ,以观察该刺激对Aβ纤维的上述机械感受特性的影响。在 42只大鼠上共记录了 5 0个Aβ类单位。逆向电刺激相邻节段皮神经后 ,60 6% (n =3 3 )的单位感受野增大 ,全部单位的感受野平均面积从 8 94± 6 5 1mm2 显著增加到 2 0 3 4± 16 17mm2 (P <0 0 1)。 81 8% (n =2 0 )的单位感受野形状从点状、圆或与身体长轴垂直的椭圆变成与身体长轴斜行或平行的椭圆。 68 0 % (n =5 0 )的单位机械感受阈值下降 ,全部单位的平均阈值从 2 3 7± 1 2 4mN降至 2 2 9± 1 2 4mN (P <0 0 5 )。上述机械感受特性的改变可持续 5 2 2 3± 9 2 7至 5 6 93± 15 76min。跨节段电刺激后 ,有 5 0 0 % (n =5 0 )的单位同时出现放电的增加 ,但该增加仅持续 1 5 2± 0 46min ,显著短于机械感受特性改变的时程 (P <0 0 1)。

初级传入纤维在发育阶段小鼠腰髓背角内生长特点的研究(英文)

本研究通过将亲脂性羰花青染料 Di I置入胚胎 12 d(E12 )到生后 3 d(P3 )小鼠背根神经节内进行追踪的方法 ,观察了初级传入纤维在小鼠腰段脊髓背角内生长过程和终止的形态特点。结果显示 ,初级传入纤维在 E13时生长至脊髓后索 ,在此停留 2d,到 E15时开始进入脊髓灰质。 E16和 E17时 ,生长至脊髓灰质内的初级传入纤维的数量增加并向腹角延伸。有相当数量的初级传入纤维进入脊髓背角深层。至 E18,背角深层的传入纤维的密度进一步增加 ,分支的状态变得更为复杂。一些分支延伸到背角浅层。在出生后 ,初级传入纤维在脊髓背角的分布特点无明显变化 ,但是在背角前层的纤维数量进一步增多。另外 ,还观察到初级传入纤维发出侧支向对侧脊髓背角的投射。这种向对侧生长的纤维侧支开始于 E16,来源于同侧背角内侧部的传入纤维。至E18和出生时 ,向对侧背角生长的侧支的来源部位增加到两个。本研究的结果表明 ,小鼠脊髓背角内初级传入纤维的板层分布模式形成于胚胎发育的晚期和出生后的早期 ,此后初级传入纤维在背角内进行进一步的精细调整 ,达到与成年时一致的状态。

猫舌快适应传入纤维止于三叉神经脊束核形成轴-树突触分析

目的:研究舌快适应传入纤维终止于三叉神经脊束核核形成的突触类型及与突触后膜-树突的关系,为神经生理学研究和神经解剖学研究提供参考。方法:采用细胞内注射,电镜连续切片技术。结果:猫舌快适应传入纤维投射到三叉神经脊束核形成的突触以中间型为主。与单纯型终扣形成突触的树突较大。与中间型终扣形成突触的树突较小。结论:各种突触类型形成的复杂联系,可能是不同感觉信息传递的形态学基础。

大鼠脊髓后角浅层初级传入C纤维内μ阿片受体的免疫组织化学定位——光镜与电镜研究

目的在光镜及电镜下观察大鼠脊髓后角内μ阿片受体（ＭＯＲ）与Ｃ纤维的关系，为进一步揭示阿片镇痛的突触机制提供形态学证据。方法根据辣椒素对初级传入神经中Ｃ纤维的特异性神经毒性，在大鼠蛛网膜下腔注射辣椒素以毁损Ｃ纤维，用免疫组织化学ＡＢＣ法比较脊髓后角内ＭＯＲ免疫反应性的变化；根据西非单叶豆同工凝集素Ｉ－Ｂ４（ｇｒｉｆｏｎｉａｓｉｍｐｌｉｃｉｆｏｌｉａｉｓｏｌｅｃｔｉｎＩ－Ｂ４，Ｉ－Ｂ４）与初级传入Ｃ纤维选择性结合的特性，应用双标（免疫）组织化学电镜技术（ＡＢＣ－纳金法）分别标记脊髓后角内Ｉ－Ｂ４结合位点和ＭＯＲ。结果大鼠蛛网膜下腔注射辣椒素３～７ｄ后，其脊髓后角Ⅰ～Ⅱ层内的ＭＯＲ免疫反应性较对照动物明显减弱；电镜下在大鼠脊髓后角浅层内观察到，ＭＯＲ不仅定位于树突内，也分布在含Ｉ－Ｂ４结合位点的Ｃ纤维终末内。无论在树突内还是在轴突终末内，ＭＯＲ主要分布在非突触部位，仅少量ＭＯＲ与突触前膜和／或突触后膜有关。结论脊髓后角中部分ＭＯＲ来源于辣椒素敏感的Ｃ纤维，外源性吗啡或内源性吗啡样物质如内吗啡肽等ＭＯＲ激动剂在脊髓后角内既可通过突触后（树突内）的ＭＯＲ发挥作用，也可通过突触前（轴突终末内）的ＭＯＲ对Ｃ纤维产生突触前?