影响黄牛和杂交受体牛胚移妊娠率的因素

1987年8月21日至9月11日于甘肃省临泽牛场进行了102例牛胚胎移植试验,报告如下: 材料与方法1.1 牛只供体为黑白花牛25头,西门达尔牛1头。均按饲养标准饲养。受体为本地黄牛,黑黄一代,西黄一代,还有1头犏牛和2头尕力巴牛(公黄牛×母犏牛的女儿)。1.2 超排采胚在发情周期9～13天,用FSH超排,连续4日,

Expression Profiles of TRAIL Receptors and Their Clinical Significance in Human Hepatocellular Carcinoma

Objective To investigate the expression profiles and their clinical significance of TRAIL receptors (TRAILR) in human hepatocellular carcinoma (HCC). Methods The expression profiles of TRAILR were determined in 60 samples from hepatocellular carcinoma, 20 from normal liver tissue and two HCC cell lines HepG2, SMMC-7721 by in situ hybridization. Results Both DR4 and DR5 were present in all HCC tissues as well as normal hepatic tissues. In contrast, 54 HCC tissues did not express DcR1 and 25 did not express DcR2. But both DcR were detectable in all of the normal liver tissues. The expression patterns of DR and DcR in HCC samples (higher DR expression level and lower DcR expression level) were quite different from those in normal tissue. DR5, DR4, and DcR2 expressed in both cell lines, while no DcR1 expression was detected. The expression level of DR was correlated with HCC differentiation and stage. The weaker expression was more commonly found in HCC with poor differentiation and late stage, while the stronger expression was more common in HCC with middle to high-differentiation and early stage. No relationship was found between DR and gender, age, negative or positive HBsAg, tumor size, grade or metastasis. Multidrug resistance cell lines expressed lower level DR. Conclusion TRAILR expression was prevalent and discrepancy of receptor types was exited in HCC. Loss of DcR1 may contribute for TRAIL therapy for HCC. Key words TRAILR - apoptosis - hepatocellular carcinoma Supported by the Major Fundation of Ministry of Health, NO. 2001–2003

Analysis on Polymorphisms of GHR Gene in Improved Hybrid Yellow Cattle Groups from Liupan Mountain Area in Ningxia

[Objective] The paper aimed to analyze the polymorphism of the growth hormone receptor （GHR） in improved hybrid yellow cattle group from Liupan Mountain area in Ningxia Autonomous Region,so as to provide technological basis for hybrid improvement. [Method] Polymerase chain reaction-restriction fragment length polymorphisms （PCR-RFLP） technology was carried out to examine polymorphisms of GHR gene of 70 individuals. [Result] The target fragment of 338 bp was amplified. The PCR product digested by restriction enzyme Alu I showed polymorphisms. The frequencies of the two genotypes （AA,BB） were 75.71% （53 individuals） and 24.29% （17 individuals）,respectively. [Conclusion] Two genotypes of GHR gene were detected in improved hybrid yellow cattle groups from Liupan Mountain area in Ningxia.

Cloning and Hybrid Identification of Banana Receptor-like Protein Kinase Gene

[Objective] This study aimed to clone and identify the banana fruit receptor-like protein kinase gene.[Method] The cDNA phage libraries of banana fruit were adopted as the experimental materials to screen positive phage libraries of banana receptor-like protein kinase gene;cloning and sequence analysis of the gene were conducted,and the banana receptor-like protein kinase gene was identified by using in situ hybridization method.[Result] In this study,a 1 698 bp long banana receptor-like protein kinase gene was cloned from banana fruit,encoding 563 amino acids.Southern hybridization result confirmed that the banana receptor-like protein kinase gene was a multiple-copy gene from banana genome.[Conclusion] The study laid the foundation for further investigating the functions of banana receptor-like protein kinase gene in fruit.

EGFR and HER2 expression in advanced biliary tract cancer

AIM:To analyze the pathogenetic role and potential clinical usefulness of the epidermal growth factor receptor(EGFR)and the human epidermal growth factor receptor 2(HER2)in patients with advanced biliary tract cancer(BTC). METHODS:EGFR and HER2 expression was studied in biopsy samples from 124 patients(51%women; median age 64.8 years),with advanced BTC diagnosed between 1997 and 2004.Five micrometers sections of paraffin embedded tissue were examined by standard, FDA approved immunohistochemistry.Tumors with scores of 2+or 3+for HER2 expression on immunochemistry were additionally tested for HER2 gene amplification by fluorescence in situ hybridisation(FISH).RESULTS:34/124 patients(27.4%)had gallbladder cancer,47(37.9%)had intrahepatic BTC and 43(34.7%)had extrahepatic or perihilar BTC.EGFR expression was examined in a subset of 56 samples. EGFR expression was absent in 22/56 tumors(39.3%). Of the remaining samples expression was scored as 1+in 12(21.5%),2+in 13(23.2%)and 3+in 9(16%), respectively.HER2 expression was as follows:score 0 73/124(58.8%),score 1+27/124(21.8%),score 2+ 21/124(17%)and score 3+4/124(3.2%).HER2 gene amplification was present in 6/124,resulting in an overall amplification rate of 5%. CONCLUSION:Our data suggest that routine testing and therapeutic targeting of HER2 does not seem to be useful in patients with BTC,while targeting EGFR may be promising.

Differential expression of autocrine motility factor receptor (AMFR) mRNA in normal and cancer cells detected by in situ hybridization

The receptor for autocrine motility factor (AMFR) is known to be involved in the process of AMF-mediated cell migration and metastasis. This paper describes the procedures of non-radioactive in situ hybridization (ISH) detection of AMFR mRNA in both paraffin-embedded surgical sections and cultured cells using either biotinylated oligonucleotide probes or digoxigenin-labeled RNA probes. The results showed that the AMFR mRNA was expressed at an enhanced level in hyperplaJstic and malignant tissues of breast and prostate cancer patient surgical specimens, indicating that the elevated AMFR expression was associated with the tissue malignancy Moreover, AMFR mRNA was detected in both normal and earcinoma cells when cultured at a subconfluent density. However, AMFR expression was inhibited in confluent normal (3T3-A31 murine fibroblast and FHs738BL human bladder) cells while it continued to express in carcinoma (J82 human bladder)and metastatic (3T3-M murine fibroblast) cells irrespective of cell density This suggested a cell-cell contact downregulation of AMFR mRNA expression in normal but not in cancer cells. The ISH data obtained in this study are closely consistent with the AMFR protein expression pattern previously reported, implying that the differential expression of AMFR gene may be regulated and controlled at the transcriptional level.

Study on the protein expression and amplification of HER2 gene in gastric cancer

Objective: The aim of the study was to investigate the human epidermal growth factor receptor 2(HER2) gene amplification and protein expression and interpretation points in the stomach mixed carcinomas. Methods: Immunohistochemistry(IHC) and fluorescence in situ hybridization(FISH) technique were used to detect HER2 gene amplification and expression of HER2 protein in 442 cases of gastric mixed carcinoma. Results: The expression rate of HER2 protein was 41.2%(182/442): the HER2 protein expression IHC 3+ extensive type in 18 cases, partial type in 21 cases, focal type in 8 cases, accounting for 10.6%(47/442); the HER2 protein expression IHC 2+ extensive type in 23 cases, partial type in 28 cases, focal type in 11 cases, accounting for 14.0%(62/442); the HER2 protein expression IHC 1+ extensive type in 27 cases, partial type in 31 cases, focal type in 15 cases, accounting for 16.5%(73/442). HER2 gene amplification rate of 442 cases was 16.1%(71/442). In 182 cases of HER2 protein positive expression, the HER2 gene cluster amplification rate was 14.8%(27/182), large granular amplification rate 11.0%(20/182), punctate amplification rate 6.0%(11/182) and high polysomy 7.1%(13/182). In 71 cases of HER2 gene amplification, there was 42 cases of HER2 protein expression IHC 3+, 22 cases of HER2 protein expression IHC 2+, and 7 cases of IHC 1+. Conclusion: HER2 detection of gastric mixed carcinoma has great heterogeneity, HER2 protein positive expression is divided into extensive type, partial type and focal type, and HER2 gene positive amplification is divided into cluster amplification, large granular amplification, punctate amplification and high polysomy. These typing of HER2 protein expression and HER2 gene amplification provide reference index to quantify for targeted therapeutic effect of anticancer drugs.

Detection of EP_1 and FP receptor mRNAs in the iris-ciliary body using in situ hybridization

OBJECTIVE: To determine the expression of E-prostanoid1 (EP(1)) and F-prostanoid (FP) receptor mRNAs in iris-ciliary bodies of the human eye using in situ hybridization. METHODS: EP(1) and FP receptor mRNAs were detected by riboprobes labeled with digoxigenin on paraffin sections of the iris-ciliary body tissue of the human eye using in situ hybridization. RESULTS: EP(1) and FP receptor mRNAs were highly expressed in blood vessels, muscles and the endothelia of the iris. EP(1) receptor hybridization signals were present in all muscle fibers of the ciliary body. Hybridization signal corresponding to FP receptor mRNA transcript was predominantly expressed in the circular muscle and in the collagenous connective tissues of the ciliary body. FP receptor mRNA was not detected in radial and longitudinal muscles. CONCLUSIONS: EP(1) and FP receptor mRNAs in human ocular tissues appear to be widely localized in the functional sites of the respective receptor agonists. Selective localization of EP(1) and FP receptor mRNAs in the circular muscles and collagenous connective tissues of the ciliary body suggests that EP(1) and FP receptors play an important role in enhancing uveoscleral outflow of aqueous humor.

Detection of EP1 and FP receptor mRNAs in the iris-ciliary body using in situ hybridization

Objective To determine the expression of E-prostanoid1 (EP 1) and F-prostanoid (FP) receptor mRNAs in iris-ciliary bodies of the human eye using in situ hybridization.Methods EP 1 and FP receptor mRNAs were detected by riboprobes labeled with digoxigenin on paraffin sections of the iris-ciliary body tissue of the human eye using in situ hybridization. Results EP 1 and FP receptor mRNAs were highly expressed in blood vessels, muscles and the endothelia of the iris. EP 1 receptor hybridization signals were present in all muscle fibers of the ciliary body. Hybridization signal corresponding to FP receptor mRNA transcript was predominantly expressed in the circular muscle and in the collagenous connective tissues of the ciliary body. FP receptor mRNA was not detected in radial and longitudinal muscles. Conclusions EP 1 and FP receptor mRNAs in human ocular tissues appear to be widely localized in the functional sites of the respective receptor agonists. Selective localization of EP 1 and FP receptor mRNAs in the circular muscles and collagenous connective tissues of the ciliary body suggests that EP 1 and FP receptors play an important role in enhancing uveoscleral outflow of aqueous humor.

p75NTR signal transduction suppressed by BFAR and p75NTR interactions

p75NTR is a low-affinity nerve growth factor receptor, which promotes cell proliferation as a positive modulator of high-affinity receptor TrkA, as well as binds with cell ligands to induce apoptosis and mediate death signals. To analyze the regulatory mechanisms of p75NTR, the present study utilized a new membrane yeast two-hybrid system to screen a human fe- tal brain cDNA library. Results identified BFAR, a novel protein that interacts with p75NTR. Interaction specificity was veri- fied by membrane yeast two-hybrid co-transformation assays, in vitro GST pull-down assays, and in vitro co-irnmunopreci- pitation assays. The fluorescent subcellular localization assay revealed that the two proteins co-localized within the cytoplasm. BFAR overexpression in PC-12 and HEK293T cells inhibited the NFnB and JNK signaling pathway, as determined with the luciferase test. Co-transfected p75NTR and BFAR in HEK293T or PC-12 cells, respectively, increased the percentage of cells in the G2/M phase, decreased the number of S-phase cells, and did not change the number of G0/Gl-phase cells.