5-羟色胺在大鼠腰骶髓后连合核和中间带外侧核参与盆腔内脏炎性痛调控的形态学研究

本研究运用荧光金(FG)逆行束路追踪与5-HT1A受体免疫荧光组织化学染色技术相结合,观察了大鼠腰骶髓后连合核(DCN)和中间带外侧核(IML)内感受盆腔内脏伤害性信息并向外侧臂旁核(LPB)发出投射的神经元呈5-HT1A受体免疫反应阳性。将FG注入一侧LPB后,可在腰骶节段(L6-S2)观察到大量的FG逆标神经元,这些FG逆标神经元主要集中于DCN和IML内,以同侧为主;5%福尔马林注入大鼠结肠后,Fos蛋白阳性神经元主要分布于腰骶髓DCN和IML,以同侧为主,在同侧脊髓背角I层、II层和深层也有少量的分布。另外,在腰骶髓DCN和IML内,还可观察到大量5-HT1A受体阳性的神经元胞体、纤维和终末,同时有部分Fos蛋白阳性的FG逆标神经元呈5-HT1A受体阳性。上述结果提示,大鼠腰骶髓DCN和IML内的5-HT能终末可能对盆腔内脏伤害性信息的传递发挥调控作用。     

巨噬细胞肿瘤疫苗抗瘤作用的实验研究

目的：探究巨噬细胞肿瘤疫苗的细胞形态与表型特征,并观察其CTL反应的诱导效果。方法：分别采用透射与扫描电子显微镜、免疫荧光染色及流式细胞术等技术对巨噬细胞肿瘤疫苗的超微结构及CD14、CD68、CD80、CD86、MHCⅡ等分子进行测定。采用生长状态良好的H22肿瘤细胞移植于接种不同肿瘤疫苗的实验小鼠,分别采用直接测量法、MTT法及比色法测定瘤重量、瘤体积、肿瘤细胞杀伤率和培养上清液LDH活性。结果：巨噬细胞肿瘤疫苗细胞表面有许多的伪足皱褶、囊泡,胞浆内有大量大小不一、形态不规则的吞噬体；CD14、CD68、CD80、CD86及MHCⅡ的阳性细胞率分别为53.90%、98.60%、26.50%、90.20%和25.40%。小鼠体内实验结果显示,巨噬细胞肿瘤疫苗接种组的肿瘤形成率、瘤体积与瘤重量明显低于对照组和石蜡诱生的巨噬细胞接种组（P均〈0.05）；巨噬细胞肿瘤疫苗接种组的成瘤率与灭活肿瘤细胞接种组无明显差异（P〉0.05）,但瘤体积与瘤重量明显低于灭活肿瘤细胞接种组（P均〈0.05）。另外,巨噬细胞肿瘤疫苗接种组的淋巴细胞自体肿瘤细胞杀伤率和培养上清液LDH活性分别高于对照组、灭活肿瘤细胞接种组和石蜡诱生的巨噬细胞接种组。结论：巨噬细胞肿瘤疫苗具备巨噬细胞的典型特征,该种细胞接种后可诱导机体产生特异性抗肿瘤免疫反应。     

孤独症广泛表型的临床调查

目的:调查孤独症先证者家系中孤独症广泛表型(BAP)及其主要表现,统计其发生率,为进一步的神经心理和遗传研究建立基础.方法:使用家系调查表和Bolton孤独症广泛表型的操作定义手册,对75例孤独症先证者的家系进行筛查,对符合BAP定义的家属进行总结和分类.结果:75例先证者的家系亲属中共有42例符合BAP 的诊断,他们以社交缺陷表现为主,其次为语言发育障碍,以行为怪异为主要表现的最少.结论:BAP在孤独症家系中的存在对遗传学的研究可能提供一条有用的途径,有必要做进一步的研究和探讨.     

Genotype-phenotype correlation in von Hippel-Lindau families with renal lesions

von Hippel-Lindau (VHL) disease arises from mutations in the VHL gene and predisposes patients to develop a variety of tumors in different organs. In the kidney, single or multiple cysts and renal cell carcinomas (RCC) may occur. Both inter- and intrafamilial heterogeneity in clinical expression are well recognized. To identify VHL-dependent genetic factors, we investigated the renal phenotype in 274 individuals from 126 unrelated VHL families in whom 92 different VHL mutations were characterized. The incidence of renal involvement was increased in families with mutations leading to truncated protein (MLTP) or large rearrangement, as compared to families with missense changes (81 vs. 63%, respectively; P=0.03). In the latter group, we identified two mutation cluster regions (MCRs) associated with a high risk of harboring renal lesions: MCR-1 (codons 74-90) and MCR-2 (codons 130-136). In addition, the incidence of RCC was higher in families with MLTP than in families with missense changes (75 vs. 57%; P=0.04). Furthermore, mutations within MCR-1 but not MCR-2 conferred genetic susceptibility to develop RCC. Overall, our data argued for a substantial contribution of the genetic change in the VHL gene to susceptibility to renal phenotype in VHL patients.     

High risk for unbalanced segregation of some reciprocal translocations: A large pedigree containing distal 4q trisomy from t(4;7)(q28;p22)

We report on a familial t(4;7)(q28;p22) with 2:2 adjacent‐1 unbalanced segregation producing duplication of 4q28→qter in multiple offspring. Within the large four‐generation pedigree, a carrier had a reproductive outcome that was approximately equal for 1) the balanced translocation, 2) normal chromosomes, and 3) viable 4q trisomy or pregnancy loss. The three individuals with chromosomal confirmation of trisomy 4q28→qter (comprising approximately 1.8% of the haploid autosomal length) had similar mental and developmental retardation, hypotonia, restricted speech, seizures, and facial anomalies but no cardiac, renal, or skeletal anomalies. It is suggested that these latter severe malformations, associated with the classic 4q2 to 3 group of anomalies, were from an imbalance outside 4q28→qter and were not necessarily related to the relatively large size of the trisomic segment. Multiple different chromosomes are reported to be rearranged with 4q in the production of distal 4q trisomy. The incidence of 4q rearrangement remains unexplained, but once it is present in a family, viability of a large trisomy in 4q seems to explain the number of affected individuals reported. © 2001 Wiley‐Liss, Inc.     

Increased spontaneous secretion of rheumatoid factor by intestinal lamina propria mononuclear cells from Crohn's disease but not ulcerative colitis patients.

Increased levels of rheumatoid factors (RF) have been observed in the serum of Crohn's disease but not ulcerative colitis patients, and have been proposed to relate to an increased state of intestinal lymphocyte activation. We have therefore examined the spontaneous in vitro secretion of RF by intestinal lamina propria mononuclear cells (MNC) isolated from specimens from control and inflammatory bowel disease (Crohn's disease, ulcerative colitis) patients. Normal intestinal lamina propria MNC spontaneously secrete rheumatoid factors of different isotypes during 14 days of in vitro culture (9.7 ng/ml IgA RF, 11.6 ng/ml IgM RF and 64.6 ng/ml IgA anti-Fc (IgG)). In matched studies intestinal MNC isolated from normal large bowel exhibited significantly greater levels of RF synthesis and secretion in vitro than normal small bowel intestinal MNC. A large increase in spontaneous RF secretion was observed from Crohn's disease intestinal MNC (21.4 ng/ml IgA RF, 21.4 ng/ml IgM RF, and 108.15 ng/ml IgA anti-Fc (IgG)), when compared with normal controls. The amount of RF secreted was dependent on the amount of inflammatory activity of the bowel specimens, from which the MNC were isolated (198.3 ng/ml of IgA anti-Fc(IgG) from involved versus 50.0 ng/ml from matched non-involved tissue). Ulcerative colitis MNC released decreased amounts of RF (7.1 ng/ml IgA RF, 6.2 ng/ml IgM RF, and 42.3 ng/ml IgA anti-Fc(IgG)). These observations using isolated intestinal MNC may explain the findings of RF changes in the sera of inflammatory bowel disease patients. Our observations support the hypothesis of a heightened state of activation in normal intestinal lamina propria MNC, which is further increased in active Crohn's disease. The dissimilarities observed between Crohn's disease and ulcerative colitis may indicate fundamental differences in disease pathophysiology and will lead to further studies exploring intestinal immunoregulatory properties of RF.     

Inflammatory malignant fibrous histiocytomas and dedifferentiated liposarcomas: histological review, genomic profile, and MDM2 and CDK4 status favour a single entity

Inflammatory malignant fibrous histiocytoma (inflammatory MFH) is a very rare tumour that occurs most often in the retroperitoneum. So far, it has been considered to be a special subtype of MFH. As it is now widely accepted that most retroperitoneal pleomorphic MFHs are dedifferentiated liposarcomas, the present study compared histological features, genomic profile (CGH analysis), and MDM2 and CDK4 status (immunohistochemistry, FISH, and quantitative PCR) in inflammatory MFHs from 12 patients and dedifferentiated liposarcomas that had an inflammatory MFH component from eight patients. Metaphase cytogenetic and FISH analyses were also performed on one inflammatory MFH. Histological review showed areas of well-differentiated liposarcoma in nine inflammatory MFHs. CGH analysis showed 12q13-15 amplification or gain in six of seven inflammatory MFHs and in seven of seven dedifferentiated liposarcomas. Immunohistochemistry showed positivity of tumour cells for MDM2 in every tumour in both groups and for CDK4 in ten and seven inflammatory MFHs and dedifferentiated liposarcomas, respectively. Metaphase cytogenetic and FISH analysis performed on one inflammatory MFH showed the presence of a supernumerary large marker chromosome and ring chromosome with high-level amplification of both MDM2 and CDK4 genes. FISH analysis on paraffin wax-embedded sections showed amplifications of MDM2 and CDK4 in seven of seven inflammatory MFHs and in seven of seven dedifferentiated liposarcomas. Quantitative PCR showed amplification of MDM2 in six and of CDK4 in seven of nine inflammatory MFHs. In conclusion, this study strongly suggests that most so-called inflammatory MFHs are dedifferentiated liposarcomas.     

CD4+ T regulatory cells from the colonic lamina propria of normal mice inhibit proliferation of enterobacteria-reactive,disease-inducing Th1-cells from scid mice with colitis

Adoptive transfer of CD4+ T cells into scid mice leads to a chronic colitis in the recipients. The transferred CD4+ T cells accumulate in the intestinal lamina propria (LP), express an activated Th1 phenotype and proliferate vigorously when exposed ex vivo to enteric bacterial antigens. As LP CD4+ T cells from normal BALB/c mice do not respond to enteric bacterial antigens, we have investigated whether colonic LP-derived CD4+ T cells from normal mice suppress the antibacterial response of CD4+ T cells from scid mice with colitis. LP-derived CD4+ T cells cocultured with bone marrow-derived dendritic cells effectively suppress the antibacterial proliferative response of CD4+ T cells from scid mice with colitis. The majority of these LP T-reg cells display a nonactivated phenotype and suppression is independent of antigen exposure, is partly mediated by soluble factor(s) different from IL-10 and TGF-beta, and is not prevented by the addition of high doses of IL-2 to the assay culture. Functionally and phenotypically the T-reg cells of the present study differ from previously described subsets of T-reg cells. The presence of T cells with a regulatory potential in the normal colonic mucosa suggests a role for these cells in the maintenance of local immune homeostasis of the gut.