促红细胞生成素对糖尿病大鼠视网膜形态的影响

目的:观察促红细胞生成素对糖尿病大鼠视网膜形态学影响.方法:将45只Wistar大鼠随机分成3组,即正常对照组、糖尿病模型组和EPO干预组,链尿佐茵素诱导糖尿病大鼠模型,6wk后行视网膜光镜、电镜观察.结果:正常组大鼠视网膜组织结构完整,视网膜分层清晰,细胞形态完好;糖尿病模型组视网膜组织结构完整,视网膜分层稍紊乱,神经节细胞层、内颗粒层及外颗粒层细胞减少、排列紊乱,神经节细胞层可见明显细胞水肿、嗜酸样变性及少量坏死;rhEPO干预组视网膜组织结构完整,视网膜分层清晰,与正常组接近.正常大鼠视网膜透射电镜下见细胞排列紧密,未出现明显的空泡变性,神经节细胞形态正常,自视杆视锥层至神经纤维层的超微结构未见明显改变;糖尿病组大鼠视网膜出现大量空泡变性,神经节细胞及内颗粒层细胞出现空泡改变,线粒体肿胀,线粒体嵴断裂、消失;rhEPO干预组大鼠视网膜在透射电镜下可见细胞排列紧密,有少量空泡改变,神经节细胞形态接近正常,内颗粒层细胞的超微结构未见明显改变.结论:EPO对早期糖尿病大鼠视网膜病变具有防治作用.     

空肠弯曲菌感染与Guillain—Barre综合征

目的：探讨空肠弯曲菌感染与Ｇｕｉｌｌａｉｎ－Ｂａｒｒｅ综合征（ＧＢＳ）发病的关系以及空肠弯曲菌感染和抗－ＧＭ１抗体与疾病严重程度的关系。方法：用ＥＬＩＳＡ方法检测８５例ＧＢＳ患者血清中的空肠弯曲菌ＩｇＧ、ＩｇＭ、ＩｇＡ抗体及抗－ＧＭ１抗体,同时从患者的大便中培养空肠弯曲菌;根据Ｈｕｇｈｅｓ报道的方法对病情轻重进行评分。结果：ＧＢＳ患者空肠弯曲菌感染率为５１．８％,抗－ＧＭ１抗体检出率为４２．４％;     

先天性巨结肠不同节段肠壁神经、平滑肌的分布及临床意义

目的 观察先天性巨结肠(HD)不同节段肠壁神经和平滑肌的病变范围,探讨先天性巨结肠根治术后肠动力功能紊乱原因及手术切除结肠范围.方法 用免疫组织化学和苏木素-伊红(HE)染色法,检测20例先天性巨结肠肠壁神经节细胞、神经纤维和平滑肌细胞病理组织学改变及分布范围.结果 巨结肠不同节段肠壁神经节细胞、神经纤维数量及突触素(Syn)、神经节细胞黏附分子(NCAM)的阳性表达,在距扩张远端8 cm虽未达到正常,但与对照组差异减小(P＞0.05).环肌层和纵肌层出现不同程度增厚,在距扩张远端8 cm仍未正常(P＜0.01).肌层出现空泡样变,与对照组比较差异无统计学意义(P＞0.05).结论 先天性巨结肠切除段肠壁神经、平滑肌层均存在病变,在距扩张段的远端8 cm处,两者病变总体缓解.在允许情况下,手术切除结肠的范围应达到或超过此范围.     

升结肠翻转术治疗先天性巨结肠症(附28例报告)

<正> 先天性巨结肠是一种常见的消化道畸形,其结肠肌间神经节细胞先天性缺如,病变肠段失去正常蠕动及排便反射的功能,其中长段型约占10％～40％,无神经节细胞段可达到降结肠及横结肠。而部分普通型患儿因就诊时间晚,近端结肠代偿性扩张肥厚严重,且肌间神经节细胞也继发变性。巨结肠根     

便秘二号方对慢传输型便秘大鼠Cajal间质细胞及C-KIT表达的影响

目的:观察便秘二号方对结肠慢传输型便秘大鼠模型的疗效,并检测其对大鼠模型结肠组织Cajal间质细胞及C-KIT表达的影响。方法:32只大鼠随机分为正常组、模型组、实验组、盐水对照组,每组8只。正常组给予正常饮食,其余3组采用大黄递增灌胃法制作慢传输型便秘大鼠模型。造模成功后,实验组予中药便秘二号方(方由当归、白术、黄芪、升麻、杏仁、枳实、肉苁蓉组成)灌胃,盐水对照组予生理盐水灌胃。30d后以活性炭灌胃法检测结肠传输功能,以免疫组化法观察结肠组织Cajal间质细胞的形态和数量,采用RT-PCR法观察C-KITmRNA表达。结果:炭末推进长度模型组明显少于正常组(P0.05),实验组明显高于盐水对照组(P0.05),推进率比较差异均有统计学意义(P0.01);实验组与盐水对照组相比,可见各肌层Cajal间质细胞数量增加,细胞形态恢复;实验组与盐水对照组的Cajal间质细胞个数比较,实验组高于对照组,差异有统计学意义(P0.01);模型组C-KIT表达阳性率明显低于正常组,实验组C-KIT表达阳性率明显高于盐水对照组。结论:中药便秘二号方可以增强慢传输型便秘大鼠的结肠蠕动功能,增加结肠组织中Cajal间质细胞的数量,改善其形态,促进C-KIT表达。     

巨结肠及类缘病诊断和治疗的临床研究

目的：探讨巨结肠类缘病的病理形态学特征，提高其诊断率。方法：对97例行巨结肠根治术后的病变肠段进行常规HE染色，观察肠神经元及神经节细胞形态和数量的变化并与正常组进行比较，并21例采用NSE和S－100蛋白免疫组化染色。结果：巨结肠类缘病（allied HD）标本的神经元与神经节细胞细胞数与正常组及巨结肠症（HD）比较差异有显著性（P＜0．01）。结论：巨结肠类缘病的诊断主要根据病理特征而定，在HWE染色基础上结合NSE及S－100蛋白免疫组织化学染色提高确诊率。     

先天性巨结肠层粘连蛋白基因及RET基因的表达研究

目的:通过对先天性巨结肠(HD)病变段层粘连蛋白(LN)转录表达的研究,探讨肠壁微环境改变对HD的形成作用以及与RET基因学说的相关性。方法:利用RT-PCR技术对HD病儿层粘连蛋白mRNA在无神经节细胞段、有神经节细胞段、正常段的表达进行检测,美国alpha9900凝胶图像分析系统判定表达强度,SPSS软件统计分析,推断LN的作用,同时检测RET基因的表达,用直线相关关系研究两者的相关性。结果:LN基因在无神经节细胞段异常高表达(P〈0.05),无神经节细胞段〉有神经节细胞段〉正常段;而RET基因的表达正好相反,无神节细胞段〈有神经节细胞段〈正常段,在无神经节细胞段明显减少,两者存在负相关关系。结论:HD无神经节细胞段LN的增高是内在的,LN增高引起肠神经细胞过早分化、定居导致无神经节细胞症的发生,可能与RET基因有一定内在联系。     

结肠冗长的特发性便秘的结肠形态病理学分析

目的 研究表现为结肠冗长的特发性便秘的结肠形态病理学改变。方法 选取14例原发性结肠冗长病例与10例不伴便秘与梗阻的结肠癌病例作为对照,通过肉眼及HE染色后显微镜观察和图像分析,比较两组的异同点。并通过定量测量和统计学分析得出显著性结果。结果 原发性结肠冗长病例外观正常,显微镜下突出的改变为5例肌间神经丛中可见巨大的神经节细胞。图像分析结果显示其粘膜下血管面积、神经丛面积和神经节细胞数,肌间神经丛面积和神经节细胞数均显著大于对照组,肌层厚度则无显著性差异。结论 原发性结肠冗长有其特征性的神经病理学改变,进一步支持了特发性便秘是一种神经系统异常性疾病的论断。     

先天性巨结肠突触素免疫组织化学研究

目的:采用突触素免疫组织化学方法,对先天性巨结肠神经肌肉连接进行研究,并探讨其与先天性巨结肠发病的关系。方法:应用免疫组化技术对20例先天性巨结肠病变肠段及10例正常结肠组织标本行突触素神经肌肉连接标记,光镜下观察其免疫反应。结果:对照组的结肠突触素免疫反应呈强阳性表达,先天性巨结肠组扩张肠段突触素免疫反应呈阳性或弱阳性表达,狭窄段肠壁突触素免疫反应呈阴性表达。结论:先天性巨结肠病变肠段同时缺乏内源性神经支配和外源性神经支配,处于完全失神经支配状态,导致原神经节细胞病变肠管功能障碍。     

空肠弯曲菌致病分子机制研究进展

空肠弯曲菌(Campylobacter jejuni)是人类的食源性病原菌,感染后主要引起急性肠炎,与人类格林-巴利综合症也有密切关系.研究表明,空肠弯曲菌致病性是多种毒力因子共同作用的结果.作者从分子生物学研究水平综述空肠弯曲菌黏附、定植、侵入、产细胞毒素、分子模拟等致病机制.     

Immunohistochemical investigations of gut hormones in the colon of patients with Hirschsprung's disease

The distributions of gut hormones in the colon of Hirschsprung's disease were investigated by the peroxidase-antiperoxidase (PAP) immunohistochemical method. Three colonic segments (ganglionic, oligoganglionic, and aganglionic) were stained by the unlabeled antibody enzyme method. The immunoreactivity of vasoactive intestinal polypeptide (VIP) was found to be reduced in the oligoganglionic and aganglionic segments. Antisera to substance P and met-enkephalin demonstrated immunoreactive cells and fibers in the ganglionic segment, whereas these cells and fibers were almost completely absent in the oligoganglionic and aganglionic segments. A similar distribution was seen for the mucosal endocrine cells with somatostatin immunoreactivity. Antisera to neurotensin, motilin, bombesin, and cholecystokinin revealed no immunoreactivity in the normal colon or the three segments. The differences in these peptides between normal and impaired colonal segments may be one of the causes of colon constriction in Hirschsprung's disease.     

双特异性单链抗体介导的CIK细胞对结肠癌的体内杀伤作用

目的 观察双特异性单链抗体介导的细胞因子诱导的杀伤细胞(CIK)细胞在体内杀伤结肠癌细胞的作用.方法 将SW1116细胞种植裸鼠后,建立裸鼠结肠癌模型,分为3组分别经尾静脉注入生理盐水、CIK细胞及CIK细胞+双特异性抗体,每周治疗1次,共4周,取出肿瘤组织称重,并计算出各组的抑瘤率.结果 在体内,CIK组和CIK+双抗组均可抑制肿瘤生长,抑瘤率分别为29.70%和65.02%,与对照组比较差异均有统计学意义(P＜0.05).CIK组和CIK+双抗组的瘤体积、瘤重、抑瘤率比较,差异均有统计学意义(P＜0.05).结论 CIK细胞本身可以抑制肿瘤细胞的生长,在加入抗CD3抗癌胚抗原(CEA)双特异性单链抗体后抑瘤作用明显增强.

Abstract：

Objective To investigate the killing effect of cytokine-induced killer cell (CIK) mediated by bispecific single-chain antibody in vivo on colon cancer cells. Methods The colon cancer model in nude mice was established with SW1116 cells. The mice were divided into 3 groups, which were injected with normal saline, CIK cells and CIK cells + bispecific antibody, respectively, once a week for 4 weeks. The tumors were removed and weighed, and the tumore inhibition rate in each group was calculated. Results The CIK cells and CIK cells with bispecific single-chain antibody could both inhibit tumor growth in vivowith the tumor inhibition rate being 29. 70% and 65.02% respectively ( P ＜ 0. 05 ). There was significant difference in tumor inhibition rate between CIK cells and CIK cells with bispecific single-chain antibody ( P ＜ 0. 05 ). Conclusion CIK cells alone can inhibit tumor cell growth, CIK cells in combination with bispecific single-chain antibody can exert more significant antitumor effect.     

Segmental aganglionosis of the appendix

Ignoring the pathologist's dictum that the appendix is not reliable for the identification of ganglion cells in suspected cases of total colonic aganglionosis, the author (K.D.A.) performed an ileostomy in a child whose appendix contained no ganglion cells. The ascending colon was later found to be ganglionic. The appendices from 36 cases of Hirschsprung's disease were examined and the number of ganglion cells in 10 high power fields was compared with 10 appendices from patients who did not have Hirschsprung's disease. There were no ganglion cells in any of the patients with total aganglionosis of the colon. Twenty-two patients with Hirschsprung's disease involving the sigmoid colon had an average of 3.0 ganglion cells per high power field (range 0.5 to 5.1). Five patients with longer segment aganglionosis had 2.2 cells per high power field (range 0 to 4.1). The index case fell into this group and was serially sectioned without finding any ganglion cells. The control group averaged 3.41 cells per high power field (range 1.4 to 5.9). There was no significant difference between the control group and the groups with less than total colonic aganglionosis. It is speculated that this case represents segmental Hirschsprung's disease with the skip area in the ascending colon.     

Mucosal neuroendocrine cell abnormalities in the colon of patients with Hirschsprung's disease.

We studied the distribution of mucosal neuroendocrine (NE) cells in the colon from 13 patients with Hirschsprung's disease (HD) and from 8 controls. Immunohistochemical studies were carried out using monoclonal and polyclonal antibodies against chromogranin A and synaptophysin (general markers of NE cells), 5-hydroxytryptamine (5-HT) (a marker of amine), peptide YY (PYY), and somatostatin (markers of neuropeptides). Chromogranin A immunoreactive cells were significantly increased in the aganglionic bowel compared with ganglionic bowel and controls (P less than .05). There was an increase in the number of synaptophysin immunoreactive cells in the aganglionic bowel compared with ganglionic bowel and controls but the results were not statistically significant. 5-HT immunoreactive cells were also significantly increased in the aganglionic bowel compared with ganglionic bowel and controls (P less than .05). The immunostaining for PYY demonstrated abundance of this NE cell type in the aganglionic bowel and this was highly significant compared with ganglionic bowel and controls (P less than .001). There was a significant increase in somatostatin immunoreactive cells in the aganglionic bowel compared with ganglionic bowel (P less than .01). The increase in neuroendocrine cells was found over the entire length of the aganglionic segment in rectosigmoid HD as well as in long-segment HD. These results demonstrating the increased levels of NE cells in the mucosa of aganglionic colon suggest that the NE cells may have a role in regulating the sustained contraction of the aganglionic intestine in HD.     

Sympathetic neurotransmitter metabolism in Hirschsprung's disease.

Tyrosine hydroxylase activity was measured in high speed supernatants obtained from full thickness segments of aganglionic and ganglionic colon of three children with Hirschsprung's disease. Tyrosine hydroxylase activity expressed as pmole DOPA/mg protein/min was 0.93 +/- 0.16 in ganglionic and 2.67 +/- 0.21 in aganglionic colon. Tyrosine hydroxylase activity in ganglionic colon rose to 2.29 +/- 0.11 following calcium stimulation (100 muM) but could not be further increased in aganglionic colon. Addition of norepinephrine (2 X 10(-4) M) to tissue homogenates inhibited tyrosine hydroxylase activity in ganglionic colon by 57 +/- 8% but only by 14 +/- 3% in aganglionic colon, suggesting that the enzyme present in aganglionic colon is insensitive to feedback inhibition by endogenous norepinephrine. The elevation of tyrosine hydroxylase activity in aganglionic colon and its insensitivity to calcium stimulation and norepinephrine inhibition is further evidence of sympathetic overactivity in the aganglionic colon and suggests a basic enzymatic abnormality in the pathogenesis of Hirschsprung's disease.     

The role of nitrergic system on the contractility of colonic circular smooth muscle in Hirschsprung's disease.

The contribution of nitrergic tone on the contractility of colonic smooth muscle in Hirschsprung's disease (HD) was investigated. METHODS: Ganglionic and aganglionic bowel specimens were taken from 8 patients with HD during pull-through operations and electrical field stimulation (EFS)-induced isometric contractions of the circular smooth muscle were recorded in vitro. Isolated circular muscle strips prepared from colonic segments of sex- and age-matched patients (n = 3) who underwent surgery for nonmotility-related colonic diseases formed the control group. Statistical analysis was performed by two way analysis of variance and unpaired Student's ttest. RESULTS: The amplitude of spontaneous rhythmic activity was lower in aganglionic segments than in ganglionic ones. The amplitudes of contractile responses were significantly greater in aganglionic segments. In ganglionic preparations, N(omega)-nitro-L-arginine (L-NNA) addition into the medium increased the contractile responses to the level of aganglionic preparations. This increase was completely blocked by L-arginine application. Neither L-NNA nor L-arginine produced any change in aganglionic segments. A relaxation phase was detected in both ganglionic and aganglionic segments. In ganglionic preparations, this relaxation phase was completely inhibited by L-NNA and restored by L-arginine, whereas no effect was detected in aganglionic ones. Responses obtained from the control group were similar to the ganglionic segments of HD patients. CONCLUSIONS: In normal colon and as well as in ganglionic segments of HD, the evoked contractile activity and relaxations are under the tonic influence of the nitrergic system. Aganglionic segments totally lack the nitrergic activity in both evoked contraction and relaxation responses, while still maintaining an inefficient relaxation capacity under unknown mechanisms.     

Does Abnormal Expression of Acetylcholine Receptors in Hirschsprung's Disease Induce Acetylcholine Esterase Positive Nerve Fibres?

OBJECTIVE: Alpha bungarotoxin (alpha-BTX) is a neurotoxin isolated from the venom of Bungarus multicinctus that binds specifically to the beta-subunits of nicotinic acetylcholine receptors (nAChR) on myotube membranes. The purpose of the present study was to investigate the distribution of alpha-BTX-sensitive nAChR in Hirschsprung's disease (HD) to understand the histopathological features of HD, especially the increase in acetylcholine esterase (AChE) positive nerve fibres. METHODS: Confocal microscopy was used to study the expression of FITC (fluorescein isothiocyanate)-alpha-BTX, anti-synaptophysin (A-SY) antibody, and anti-neurofilament (A-NF) antibody to determine the distribution of nAChR and ganglion cell and nerve fibres in colon specimens from five cases of HD and three normal controls. RESULTS: Quantitative assessment of the immunoreactivity of colonic muscle and colonic mucosal epithelium from an aganglionic segment of HD bowel demonstrated markedly increased nAChR compared with colonic muscle and colonic mucosal epithelium from a ganglionic segment of HD bowel and normal bowel (p < 0.0001, respectively), both of which have only a few positive nAChR. In colonic muscle from aganglionic and transitional segments of HD, there were many nAChR around hypertrophic nerve trunks identified by A-NF and A-SY staining. CONCLUSION: We suggest that abnormal expression of nAChR in HD might be implicated in causing gastrointestinal dysmotility because of their localization around hypertrophic nerve trunks.     

Increased expression of the epidermal growth factor receptor on human colon carcinoma cells

Epidermal growth factor (EGF) is a small polypeptide hormone that promotes the growth of cells in culture and elicits the differentiation of epithelial tissues in vivo. The effect of EGF is mediated by a transmembrane receptor that is expressed in increased amounts on some tumor cells. We have used a monoclonal antibody to the EGF receptor to detect increased expression of the receptor on human colon carcinoma cells. All eight of the moderately well-differentiated colon carcinoma cell lines tested and several frozen colon carcinoma tissue sections showed increased expression of the EGF receptor, while five poorly differentiated colon carcinoma cell lines and normal colon tissue sections did not. Increased expression of the EGF receptor on moderately well-differentiated colon carcinoma cells but not on poorly differentiated colon carcinoma cells was also demonstrated by western transfer and iodine 125-labeled EGF binding assays. Increased expression of the EGF receptor on moderately well-differentiated colon carcinoma cells seems to be a useful marker for the differentiation of human colon carcinoma cells. In addition, it might provide a site for adjuvant hormonal therapy or immunotherapy.