胶原纤维、TRPV1在直肠黏膜内脱垂组织中免疫化学定位和表达研究

目的:探讨胶原纤维、瞬时感受器电位香草酸受体1(TRPV1)在直肠黏膜内脱垂组织中免疫化学定位和表达的意义。方法:将行手术治疗的直肠黏膜内脱垂患者40例纳入观察组,再抽取同期健康体检者40例纳入对照组。均经结肠镜检查后,评估直肠胶原纤维和TRPV1免疫化学定位及表达情况。结果:1)胶原纤维表达中,Masson染色和EVG染色均出现结构纤细状况,前者伴有数目稀疏,后者伴有分布广和破碎状、结构紊乱性弹性纤维,且胶原纤维面积比明显<健康体检者,数据比较差异有统计学意义(P<0.05)。2)TRPV1表达中,阳性区呈棕褐色,多见黏膜腺上皮细胞,和健康体检者相比免疫反应阳性增强、图像分析增强5.28倍,数据比较差异有统计学意义(P<0.05)。结论:对直肠黏膜内脱垂患者,黏膜层胶原纤维、TRPV1反应性增强是引起直肠高敏感性的前兆,可能是导致机体排便功能障碍的首要机制。

嫁接早期IAA在西葫芦/南瓜嫁接面处薄壁细胞和维管分子中的免疫细胞学定位

采用免疫胶体金法对IAA在西葫芦/南瓜离体茎段嫁接早期发育时期在嫁接面处的分布进行了超微结构水平的定位。电镜观察表明:在嫁接面处的薄壁细胞中IAA主要定位于细胞核、质体、内质网等细胞器上。在高尔基体、线粒体、细胞壁和液泡中,未发现胶体金颗粒的标记。在分化中的管状分子中,胶体金颗粒位于次生壁上和细胞质中。在筛分子分化过程中,IAA主要定位于筛板、筛孔和细胞质中。在伴胞中有较高的金颗粒密度。对于IAA在嫁接体维管分子分化过程中的作用进行了讨论。

Immunohistochemical Analysis of IAA in Anthers of the Photoperiod-sensitive Genic Male-sterile Rice

The immunohistochemical localization of IAA and the comparison of their relative levels were carried out for the first time in the anthers of Nongken 58S and its wild type Nongken 58 (Oryza sativa subsp. japonica) after long_day and short_day treatments. The distribution of free_IAA in anthers and its dynamic variation could be reflected by this method. The results showed that the IAA level in the anthers of Nongken 58S after long_day treatment was much lower than that in short_day_treated Nongken 58S and those in wild type Nongken 58 in five stages from pistil and stamen primordia formation to late uninucleate stage. The possible reasons for IAA deficiency in Nongken 58S_LD anthers and its relationship with fertility alteration were also discussed.

Immunohistochemical Location of Six Kinds of Endocrine Cell from Digestive Tract of Misgurnus anguillicaudatus

[Objective] The study was to immunohistochemical localize the endocrine cells from digestive tract of Misgurnus [ Method ] Using six gastrointestinal hormone antisera, the endocrine cells from digestive tract of M. anguillicaudatus was localized. [ Result ]The 5-hydroxytryptamine immunoreactive（5-HT-IR） cells distribute in oesophagus, foregut and midgut; the distribution density was determined to be forepart of foregut 〉 oesophagus and hindpart of foregut 〉 gut, and the differences in the three density gradients reached significant level. Like PP-IR, SS-IR cells were observed mostly in oesophagus, followed by hindpart of foregut, least in forepart of foregut, but never found in gut and hindgut. The three kinds of immunocompetent cells Gas-IR, Glu-IR and SP-IR were not detected in each part of digestive tract. [ Conctusion] This study may provide basic data for studying the nutritional and digestive physiology, as well as the preparation of meridic diets for M. anguillicaudatus.

FoxO4 is the main forkhead transcriptional factor localized in the gastrointestinal tracts of pigs

Forkhead box （Fox） proteins play critical roles in the regulation of differentiation, proliferation, immunity and aging of cells. Most studies on Fox proteins are limited to cultured cells and rodent. The aim of the current study is to detect by immunohistrochemistry whether FoxO1, FoxO3a and FoxO4 proteins are localized in the stomach and intestine of the pig. The results showed that FoxO4 exists in the mucosa in all parts of the stomach and intestine; FoxO3a exists mainly in the lamina propria and muscularis of some parts. However, FoxOl is not detectable in all parts of the stomach and intestine. Collectively, the results of the present study indicate that there exists a distinct expression pattern of Fox proteins, and that FoxO4 is a primary forkhead transcriptional factor localized in the gastrointestinal tracts of the pig.

Gene expression profiling and endothelin in acute experimental pancreatitis

AIM:To analyze gene expression profiles in an experimental pancreatitis and provide functional reversal of hypersensitivity with candidate gene endothelin-1 antagonists.METHODS:Dibutyltin dichloride(DBTC) is a chemical used as a polyvinyl carbonate stabilizer/catalyzer,biocide in agriculture,antifouling agent in paint and fabric.DBTC induces an acute pancreatitis flare through generation of reactive oxygen species.Lewis-inbred rats received a single i.v.injection with either DBTC or vehicle.Spinal cord and dorsal root ganglia(DRG) were taken at the peak of inflammation and processed for transcriptional profiling with a cDNA microarray biased for rat brain-specific genes.In a second study,groups of animals with DBTC-induced pancreatitis were treated with endothelin(ET) receptor antagonists [ET-A(BQ123) and ET-B BQ788)].Spontaneous pain related mechanical and thermal hypersensitivity were measured.Immunohistochemical analysis was performed using anti-ET-A and ET-B antibodies on sections from pancreatic tissues and DRG of the T10-12 spinal segments.RESULTS:Animals developed acute pancreatic inflammation persisting 7-10 d as confirmed by pathological studies(edema in parenchyma,loss of pancreatic architecture and islets,infiltration of inflammatory cells,neutrophil and mononuclear cells,degeneration,vacuolization and necrosis of acinar cells) and the painrelated behaviors(cutaneous secondary mechanical and thermal hypersensitivity).Gene expression profile was different in the spinal cord from animals with pancreatitis compared to the vehicle control group.Over 260 up-regulated and 60 down-regulated unique genes could be classified into 8 functional gene families:circulatory/acute phase/immunomodulatory;extracellular matrix;structural;channel/receptor/transporter;signaling transduction;transcription/translation-related;antioxidants/chaperones/heat shock;pancreatic and other enzymes.ET-1 was among the 52 candidate genes upregulated greater than 2-fold in animals with pancreatic inflammation and visceral pain-related behavior.Treatments with the ET-A(BQ123) and ET-B(BQ-788) antagonists revealed significant protection against inflammatory pain related mechanical and thermal hypersensitivity behaviors in animals with pancreatitis(P < 0.05).Open field spontaneous behavioral activity(at baseline,day 6 and 30 min after drug treatments(BQ123,BQ788) showed overall stable activity levels indicating that the drugs produced no undesirable effects on normal exploratory behaviors,except for a trend toward reduction of the active time and increase in resting time at the highest dose(300 μmol/L).Immunocytochemical localization revealed that expression of ET-A and ET-B receptors increased in DRG from animals with pancreatitis.Endothelin receptor localization was combined in dual staining with neuronal marker NeuN,and glia marker,glial fibrillary acidic protein.ET-A was expressed in the cell bodies and occasional nuclei of DRG neurons in na ve animals.However,phenotypic expression of ET-A receptor was greatly increased in neurons of all sizes in animals with pancreatitis.Similarly,ET-B receptor was localized in neurons and in the satellite glia,as well as in the Schwann cell glial myelin sheaths surrounding the axons passing through the DRG.CONCLUSION:Endothelin-receptor antagonists protect against inflammatory pain responses without interfering with normal exploratory behaviors.Candidate genes can serve as future biomarkers for diagnosis and/or targeted gene therapy.

The Significance of Minimally Invasive Core Needle Biopsy and Immunohistochemistry Analysis in 235 Cases with Breast Lesions

OBJECTIVE To evaluate core needle biopsy (CNB) as a mini-mally invasive method to examine breast lesions and discuss theclinical significance of subsequent immunohistochemistry (IHC)analysis.METHODS The clinical data and pathological results of 235 pa-tients with breast lesions, who received CNB before surgery, wereanalyzed and compared. Based on the results of CNB done beforesurgery, 87 out of 204 patients diagnosed as invasive carcinomawere subjected to immunodetection for p53, c-erbB-2, ER and PR.The morphological change of cancer tissues in response to chemo-therapy was also evaluated.RESULTS In total of 235 cases receiving CNB examination, 204were diagnosed as invasive carcinoma, reaching a 100% consistentrate with the surgical diagnosis. Sixty percent of the cases diag-nosed as non-invasive carcinoma by CNB was identified to havethe presence of invading elements in surgical specimens, and simi-larly, 50% of the cases diagnosed as atypical ductal hyperplasia byCNB was confirmed to be carcinoma by the subsequent result ofexcision biopsy. There was no significant difference between theCNB biopsy and regular surgical samples in positive rate of im-munohistochemistry analysis (p53, c-erbB-2, ER and PR; P >0.05).However, there was significant difference in the expression rate ofp53 and c-erbB-2 between the cases with and without morphologi-cal change in response to chemotherapy (P < 0.05). In most caseswith p53 and c-erbB-2 positive, there was no obvious morphologi-cal change after chemotherapy.CONCLUSION CNB is a cost-effective diagnostic method withminimal invasion for breast lesions, although it still has some limi-tations. Immunodetection on CNB tissue is expected to have greatsignificance in clinical applications.

Studies on Physiological and Biochemical Properties of Female Specific Serum Protein and Its Immunocytochemical Localization in Carassius auratus cuvieri(Temminck & Schlegel)

Female-specific serum protein(FSSP)is normally present in the sera of female fish,but it is notfound in males.However,estradiol benzoate(EB)was found to induce the appearance of FSSP inmale fish and immature female fish.Massive doses of FSSP caused a rapid increase in FSSP.Ultrastructural examination of the liver indicated an extensive proliferation of the rough endoplasmicreticulum and a decrease in glycogen and lipid droplets after EB injection.A preparative PAGE for isolating highly purified FSSP from the serum of EB-treated fish,Carassiusauratus cuvieri(Temminck & Schlegel),is described.Purified FSSP obtained from EB-induced O-crucian fish has a molecular weight of 466,000±4,000(n=10),while that in mature female serumis 480,000±40,000(n=2).FSSP appears to be a dimer,with the size of the possible monomerbeing 240,000±8,000(n=6).SDS-PAGE on gradient gels indicated that sera from males given multiple(12)injections of EBcontain a main band with a molecular weight of 147,000±6,000(n=6).However,the same serumsamples provided three bands of protein on the PAGE gels.Antiserum was raised against the electrophoretically purified FSSP.The resulting antibody formeda single,continuous precipitation line with sera from EB-treated males and vitellogenic females,butnot with that from normal males.lmmunocytochemistry(PAP method)was used to locate FSSP in the liver and ovary of maturefemales and the liver of EB-treated males.Strongly positive particles were found clustered in groupsaround liver cell nuclei under light microscopy,and the yolk granules in the oocytes were also filledwith positive particles.