乙酰胆碱受体α亚单位125～147段多肽致实验性自身免疫性重症肌无力

目的　用多肽免疫动物建立实验性自身免疫性重症肌无力动物模型。方法　合成电鳗乙酰胆碱受体α亚单位Tα1 2 5～ 1 4 7片段 ,并用其免疫接种长耳白兔 ,观察接种动物的临床症状、抗体滴度及神经电传导功能变化。结果　实验兔强化接种后第 2～ 5天渐出现肌无力症状 ,新斯的明试验阳性。接种免疫原组抗多肽Tα1 2 5～ 1 4 7抗体吸光度 (0 745 8± 0 1 681 )远高于对照组 (0 1 2 79± 0 0 1 73 ,P <0 0 5) ,3、5、1 0Hz下重复神经电刺激实验 (RNS)动作电位衰减率和单纤维肌电图检查颤抖值 (MCD)均较对照组明显延长 (P <0 0 5)。结论　Tα1 2 5～ 1 4 7能作为免疫原诱导动物产生重症肌无力模型     

Anorexic action of a new potential neuropeptide Y antagonist [ -Tyr, -Thr]-NPY (27–36) infused into the hypothalamus of the rat

Neuropeptide Y (NPY) produces a vigorous feeding response in several species when it is injected into hypothalamic structures involved in eating behavior. The purpose of this study was to determine whether a unique carboxy terminal fragment of NPY would alter the pattern of eating induced in the rat either by NPY injected into the hypothalamus or by a 24-h period of food deprivation. In this case, two -tyrosine residues and one t.-threonine residue of the NPY_27–36 fragment were transformed to their D-conformation to produce [ -Tyr^27,36, -Thr³²]-NPY (27–36), i.e., D-NPY_27–36. Guide cannulae for microinjection were implanted stereotaxically just dorsal to the paraventricular nucleus (PVN) or ventromedial hypothalamus (VMH) of 24 adult male Sprague-Dawley rats. Following postoperative recovery, a microinjection of artificial CSF or 1.1 jig or 3.3 μg of a peptide was made directly into the PVN or VMH as follows; native NPY; D-NPY_27–36; or [L-Tyr^27,36 L-Thr³²]-NPY (27–36), i.e., L-NPY_27–36. Food intakes were measured at intervals of 0.25, 0.5, 1.1, 2.0, 4.0, and 24 h. When D-NPY_27–36 was microinjected at NPY reactive sites in the PVN or VMH of the rat 15 min before a similar microinjection of NPY, the intense eating response induced by the peptide was reduced significantly. Not only was the effect dose dependent, but D-NPY_27–36 also augmented the latency to feed. A mixture of the two doses of NPY and DNPY_27–36 injected at the same hypothalamic loci did not attenuate the intake of food but tended to enhance the feeding response in the rats. After the rats were deprived of food for 24 h, D-NPY_27–36 microinjected in the same hypothalamic sites similarly caused a dose-dependent suppression of normal feeding behavior. However, the CSF control vehicle and L-NPY_27–36 microinjected in the PVN or VMH were without effect on the pattern of eating. Further, D-NPY_27–38 injected in the same hypothalamic sites affected neither body temperature nor water intakes of the rats significantly. These results demonstrate that the D substitution of this C-fragment of the NPY molecule, i.e., D-NPY_27–36, serves to inhibit feeding evoked in the rat by hypothalamic NPY as well as the natural eating response to food deprivation. Thus, the D-NPY_27–36 molecule may act as an antagonist at one or more subtypes of the NPY receptor in the brain of the rat.     

肺癌组织心钠素、血清降钙素含量测定及T淋巴细胞亚群浸润程度结果分析

为了解４种不同组织学类型肺癌组织心钠素（ＡＮＰ），血清降钙素（ＣＴ）含量与Ｔ淋巴细胞亚群浸润相关程度，用放免分析法及免疫组化ＡＢＣ法测定５８例肺癌组织ＡＮＰ血清ＣＴ含量Ｔ３＋，Ｔ４＋，Ｔ８＋淋巴细胞浸润程度。结果：５８例肺癌组织ＡＮＰ，ＣＴ含量均明显高于对照组（Ｐ＜０．０１）。肺癌不同组织学类型ＡＮＰ，ＣＴ含量均高于对照组（Ｐ＜０．０１）且有不同程度相关。肺癌相同组织学类型Ｔ３＋，Ｔ４＋，Ｔ８＋／浸润程度不同，以Ｔ３＋淋巴细胞最为明显。肺癌组织ＡＮＰ，血清ＣＴ含量与Ｔ３＋，Ｔ４＋，Ｔ８＋淋巴细胞浸润程度之间有不同程度相关。表明肺癌组织ＡＮＰ，ＣＴ及Ｔ淋巴细胞浸润之间有一定内在联系。     

Reversed-phase liquid chromatography of the opioid peptides. 3. Development of a microanalytical system for opioid peptides involving microbore liquid chromatography, post-column derivatization and laser-induced fluorescence detection

A microanalytical system has been developed for the determination of peptides in small samples. Naphthalene-2,3-dicarboxaldehyde-beta-mercaptoethanol (NDA-BME) was used as the labelling reagent system as an alternative to NDA-cyanide (NDA-CN) because of the faster labelling when CN was replaced by a thiol. The fluorescence characteristics of the NDA-thiol adducts, N-substituted 1-alkylthiobenz[f]isoindoles (TBIs), were found to be different from the previously described cyanobenz[f]isoindole (CBIs) adducts formed by the reaction of primary amines with NDA-CN. The excitation maximum of the TBI adducts was at 460 nm, which was closer to the 457.9 nm argon-ion laser line, than the 440-nm maximum of the CBI adduct. The limit of detection for leucine enkephalin was 36 fmol (S/N = 3) and linearity was proven for greater than 2 orders of magnitude, from 45 fmol to 9 pmol for an injection volume of 60 nl. The detectability was limited by the high background noise produced by the post-column derivatization system. The utility of the system was demonstrated for the analysis of methionine enkephalin and its potential oxidation products, using leucine enkephalin as a suitable internal standard.     

131I-层粘连九肽的合成及在荷乳腺癌裸鼠中的显像和分布

目的用１３１Ｉ层粘连九肽进行荷乳腺癌裸鼠的肿瘤受体显像及其生物分布研究。方法以固相法合成层粘连九肽，经ＨＰＬＣ纯化，用氯胺Ｔ法进行１３１Ｉ标记。标记物以ＳｅｐｈａｄｅｘＧ１０纯化。尾静脉注入荷乳腺癌裸鼠体内，分别在１、２、３、４、５、６小时进行肿瘤显像，划出感兴趣区，测其Ｔ／Ｂ值。静脉给药后分别在３、６小时观察生物分布，计算其每克组织摄取注射剂量百分数（％ＩＤ／ｇ）和肿瘤与非肿瘤的放射性比值（Ｔ／ＮＴ值）。结果化学合成的层粘连九肽经ＨＰＬＣ、ＦＡＢＭＳ纯化鉴定，Ｍ＋１＝９６６３。尾静脉注射后４～５小时肿瘤部位有明显浓聚，Ｔ／Ｂ值达４６２。肿瘤／肌肉的Ｔ／ＮＴ值在６小时为３４５。结论１３１Ｉ层粘连九肽有希望用于诊断人体乳腺癌     

Behavioral effects of melanocyte stimulating hormone release-inhibiting factor-1 (MIF-1)

Consideration of the isolation, structure, localization, and behavioral effects of melanocyte stimulating hormone release inhibiting factor (MIF-1) is followed by a review of its opiate antagonistic and clinical effects. Evidence pertaining to various hypotheses offered in explanation of these behavioral effects is examined and evaluated. It is concluded that MIF-1 affects behavior in many instances with possible antagonistic effects as well as clinical possibilities.     

Substance P-immunoreactive processes on 5HT/SP-immunoreactive medullary cells

Two-color immunoperoxidase staining has been used to localize substance P (SP)-immunoreactive processes on neurons in the caudal medulla that exhibited both serotonin (5HT)- and SP-immunoreactivity (5HTI/SPI cells). The punctate SP-immunoreactive processes were present on 5HTI/SPI cells in the raphe nuclei and ventral reticular formation. This close association suggests that activity in bulbospinal 5HT/SP pathways, which can influence sympathetic preganglionic neurons, may be affected by the release of SP in the brainstem.     

Toll-like receptors and antimicrobial peptides expressions of psoriasis: correlation with serum vitamin D level

To evaluate the association of Toll-like receptors (TLRs), antimicrobial peptides (AMPs) and vitamin D receptors (VDRs) in psoriasis, lesional (PP) and perilesional skin (PN) from psoriasis, atopic dermatitis (AD) patients and healthy controls (NN) were studied by immunohistochemistry. Compared with PN, AD and NN skin, dysregulated expression of TLRs, AMPs and VDR was detected in PP skin. Noteworthy, our results showed altered correlation between TLR2 and VDR expression in PP and PN skin. Human beta defensin 2 (HBD2) and cathelicidin (LL-37) expressions in the PP skin were higher in serum vitamin D sufficient (VDS) groups than serum vitamin D deficient (VDD) groups. Negative correlation was found between TLR2 and VDR expression in the PP skin of VDD groups. However, positive correlation was noted in the PP skin of VDS groups. Based on the present results, therapies targeting the activity of TLRs, AMPs and vitamin D, including modulation of the TLR-VDR pathways, might provide new therapeutic approaches to the psoriasis and other inflammatory skin diseases.     

HLA class II influences humoral autoimmunity in patients with type 2 autoimmune hepatitis

BACKGROUND/AIMS: Type 2 autoimmune hepatitis (AIH) is characterized by the presence of anti-liver kidney microsome (anti-LKM-1) and/or anti-liver cytosol type 1 (anti-LC1) autoantibodies. However, the correlation between these autoantibodies and the genetic background has not been studied. METHODS: Frequencies of HLA class II alleles were compared between the 60 Caucasian children with type 2 AIH and 313 control subjects. The anti-LKM1 antibody reactivity directed against antigenic sites of CYP2D6 was analysed by ELISA. RESULTS: HLA-DQB1 *0201 allele was found to be the primary genetic determinant of susceptibility to type 2 AIH by conferring the highest odd-ratio (OR = 6.4). HLA-DRB1 *03 allele was significantly increased (P < 0.0001) among patients with both anti-LKM1 and anti-LC1 autoantibodies as well as in those with only anti-LC1(+) compared to those with anti-LKM1(+) alone. In contrast, HLA-DRB1 *07 allele was significantly associated (P < 0.0001) with anti-LKM1(+) alone compared to groups with both anti-LKM and anti-LC1 or with LC1+ alone. Children with the DRB1 *07 allele develop anti-LKM1 autoantibodies having a more restricted specificity (2 epitopes) than to those having HLA-DRB1 *03 allele (5 epitopes). CONCLUSIONS: The HLA-DR locus is involved in autoantibody expression, while the DQ locus appears to be a critical determinant for the development of type 2 AIH.     

抗核抗体和抗环瓜氨酸肽抗体在风湿性疾病中的应用

目的探讨抗核抗体（ANA）和抗环瓜氨酸肽（CCP）抗体在风湿性疾病中的敏感性及相关性。方法收集浏阳市人民医院2010年9月至2013年2月433例已确诊为风湿性疾病患者和60例健康体检人员的血清．采用间接免疫荧光试验（IIFA）检测ANA,判断荧光核型,免疫印迹试验（UA）抗核抗体谱（ANAs）12项检测特异性抗体,胶体金法测定抗CCP抗体,并对结果进行相关分析。结果（1）在433例风湿性疾病患者血清标本中,ANA核型阳性共258例．其中系统性红斑狼疮阳性率为83．03％（137／165）,类风湿关节炎（RA）阳性率为26．85％（40／149）,原发性干燥综合征阳性率为64．15％（34／53）,混合结缔组织病阳性率为[00．00％（20／20）,多肌炎／皮肌炎阳性率为33．33％（8,24）,进行性系统性硬化症阳性率为86．36％（19／22）;健康对照组阳性率为0．00％。（2）抗双链DNA（ds—DNA）抗体、抗史密斯（Sm）抗体、抗核糖体P蛋白抗体（ARPA）、抗组蛋白抗体和核小体抗体阳性标本中均质型所占比例最高：抗尿嘧啶-1低相对分子质量核糖核蛋白（抗U1．nRNP）抗体、抗干燥综合征抗原A（SS—A）抗体、抗SS．B抗体阳性标本中,斑点型所占比例最高;抗硬皮病-70（Sel-70）抗体主要见于均质核仁混合型;抗组氨酰．tRNA合成酶（Jo．1）抗体阳性标本中,胞浆型所占比例最高;抗着丝点B（CENPB）抗体则全为着丝点阳性;（3）在175例IIFA．ANA阴性（IIFA—ANA-）标本中,UA．ANAs阳性（LIA—ANAs-）1例;（4）433例风湿性疾病患者中,抗CCP抗体阳性82例,其中RA组阳性率为51．68％（77／149）,非RA组阳性率为1．76％（5／284）。结论ANAs中抗ds．DNA抗体、抗Jo．1抗体、抗CENPB抗体与ANA的荧光模式和风湿性疾病类型有一定关联：ANA核型以斑点型所占比例最高;IIFA—ANA-者检出LIA．ANAs-的可能性较小;抗CCP抗体对RA的诊断具有较好的灵敏度和特异性,是诊断RA的特异性血清学标志。