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Uniqueness of the Gossypium mustelinum Genome Revealed by GISH and 45S rDNA FISH

Journal of Integrative Plant Biology 2013年第7期55卷 654-662页

作者： Qiong Wu Fang Liu Shaohui Li Guoli Song Chunying Wang Xiangdi Zhang Yuhong Wang David Stelly Kunbo Wang Cotton Research Institute the Chinese Academy of Agricultural Sciences/National Key Laboratory of Cotton Biology Haikou Experimental Station Chinese Academy of Tropical Agricultural Sciences Department of Soil and Crop Sciences Texas A&M UniversityCollege Station

Gossypium mustelinum （（AD）4） is one of five disomic species in Gossypium. Three 45S ribosomal DNA （rDNA） loci were detected in （AD）4 with 45S rDNA as probe, and three pairs of brighter signals were detected with genomic DNA （gDNA） of Gossypium D genome species as probes. The size and the location of these brighter signals were the same as those detected with 45S rDNA as probe, and were named GISH-NOR. One of them was super-major, which accounted for the fact that about one-half of its chromosome at metaphase was located at chromosome 3, and other two were minor and located at chromosomes 5 and 9, respectively. All GISH-NORs were located in A sub-genome chromosomes, separate from the other four allopolyploid cotton species. GISH-NOR were detected with D genome species as probe, but not A. The greatly abnormal sizes and sites of （AD）4 NORs or GISH-NORs indicate a possible mechanism for 45S rDNA diversification following （AD）4 speciation. Comparisons of GISH intensities and GISH-NOR production with gDNA probes between A and D genomes show that the better relationship of （AD）4 is with A genome. The shortest two chromosomes of A sub-genome of G. mustelinum were shorter than the longest chromosome of D sub-genome chromosomes. Therefore, the longest 13 chromosomes of tetraploid cotton being classified as A sub-genome, while the shorter 13 chromosomes being classified as D sub-genome in traditional cytogenetic and karyotype analyses may not be entirely correct.

关键词： Chromosomes cotton Gossypium mustelinum in situ hybridization nucleolar organizer region genome.

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Effect of agonist on gamma-aminobutyric acid B receptor subunit expression in the hippocampus of epileptic rats

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Neural Regeneration Research 2009年第8期4卷 577-582页

作者： Jihong Tang Shiyao Bao Siming Hu Zhilin Zhang Department of Neurology Children's Hospital Affiliated to Soochow University Suzhou 215003 Jiangsu Province China

BACKGROUND： An amino acid imbalance has been considered to be responsible for epilepsy pathogenesis. Gamma-aminobutyric acid-B receptor （GABABR） inhibits voltage-sensitive calcium ion channels and GABA or glutamic acid （Glu） neurotransmitter release, which promotes or inhibits onset and development of epilepsy. OBJECTIVE： To explore the effect of baclofen on GBRla and GBR2 mRNA expression in the hippocampus of epileptic rats following kainic acid （KA） induction, and to study the adaptability of GABABR subunits. DESIGN, TIME AND SETTING： A randomized, controlled, animal experiment based on molecular biology was performed at the Laboratory Research Center of Second Hospital Affiliated to Soochow University from November 2005 to March 2006. MATERIALS： KA was provided by Sigma, USA. in situ hybridization detection kit of GBRla and GBR2 was provided by Wuhan Boster Biological Technology, China. GABABR agonist （baclofen） was provided by Sigma, USA. METHODS： Forty-four epileptic rats were randomly allocated to epileptic （n = 28） and drug intervention （n = 16） groups. The epileptic group was further divided into post-epileptic subgroups at different time points： 6, 12 hours, 1, 3, 7, 15, and 30 days （n = 4）. The drug intervention group was further divided into intervention controls subgroups at various time points： 6 hours, 1 day, and 3 days （n = 4）. Four additional rats were considered the normal control group and not modeled, but were injected with saline in the hippocampal CA3 region. MAIN OUTCOME MEASURES： GBRla and GBR mRNA expression was detected in the right hippocampal CA1, CA3, and dentate gyrus （DG） areas of the control, epileptic, and interference groups at various time intervals according to in situ hybridization results. RESULTS：（1） During the early stage of epilepsy （6 and 12 hours）, GBRla and GBR2 mRNA expression was decreased, and expression was less than the control group at one day after KA induction （P 〈 0.05）. mRNA expression was increased in the DG, bu

关键词： temporal lobe epilepsy GABAB receptor hippocampus baclofen in situ hybridization

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Identification and Expression of Floral Organ Homeotic Genes from Alpinia oblongifolia (Zingiberaceae)

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Journal of Integrative Plant Biology 2009年第2期51卷 155-166页

作者： Yong-Mei Xia Xue-Mei Gao Qing-Jun Li Xishuangbanna Tropical Botanical Garden the Chinese Academy of Sciences Menglun Mengla Yunnan 666303 China

Current understanding of the classical ABC model of floral development has provided a new set of characters to evaluate floral evolution. However, what is still lacking is a clear assessment of this genetic program across monocots. Here, to investigate the evolution of members of class A and B genes in monocots, we report the sequence characteristic and transcript expression of three new MADS-box genes in Alpinia oblongifolia Hayata. Sequence and phylogenetic analysis reveals that these genes are FUL-like and AP3-1ike. Therefore, they were termed AoFL1, AoFL2 and AoAP3. AoFL1 contains the FUL motif, but AoFL2 lacks this motif. Their expression revealed by in situ hybridization may reflect the ancestral function of FUL-like genes in the specification of inflorescence and floral meristems. The AoAP3 gene contains two conserved motifs, the PI-derived and paleoAP3 motifs. The AoAP3 transcripts located to the corolla and stamen, and hybridization signals were detected in the central whorl. These expression patterns suggest that the functions of homologous organ identity genes are diversified in A. oblongifolia. The implications of these findings on the conservation of homologous gene function are discussed.

关键词： ABC model floral organ identity in situ hybridization MADS-box genes phylogenetic analysis.

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Identification and localization of two sensory neuron membrane proteins from Spodoptera fitura （Lepidoptera： Noctuidae）

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Insect Science 2015年第3期22卷 399-408页

作者： Jin Zhang Yang Liu William B. Walker Shuang-Lin Dong Gui-Rong Wang State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing Education Ministry Key Laboratory of Integrated Management of Crop Disease and Pests College of Plant Protection Nanjing Agricultural University Nanjing China Swedish University of Agricultural Sciences Department of Plant Protection Biology Chemical Ecology Research Group Alnarp Sweden

Sensory neuron membrane proteins （SNMPs）, which are located on the dendritic membrane of olfactory sensory neurons （OSNs）, are proposed to be associated with odor reception in insects. Recent studies have demonstrated that SNMP1 is essential for electrophysiological responses of OSNs to the sex pheromone, cis-vaccenyl acetate （cVA） in Drosophila melanogaster. To investigate the function ofLepidoptera SNMPs, we cloned two SNMP genes, SlituSNMP1 and SltiuSNMP2, from Spodoptera litura （Lepidoptera： Noctuidae）. Sequence alignment and phylogenetic analysis showed that both genes bear the general characteristics of SNMPs, including six conserved cysteine residues and two transmembrane domains. Further expression profile experiments showed that SIituSNMP1 is mainly expressed in the antenna, while SlituSNMP2 is broadly expressed in various tissues. By in situ hybridization experiments, it was found that SlituSNMP1 expressing cells are surrounded by the SlituSNMP2 expressing cells in the pheromone sensitive sensilla, suggesting different fimctions ＆the two SNMPs in insect olfaction.

关键词： in situ hybridization Spodoptera litura sensory neuron membrane proteins (SNMPs)

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Clinicopathological features and prognosis assessment of extranodal follicular dendritic cell sarcoma

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World Journal of Gastroenterology 2010年第20期16卷 2504-2519页

作者： Li, Lan Shi, Yong-Hong Guo, Zhi-Juan Qiu, Tian Guo, Lei Yang, Hong-Ying Zhang, Xun Zhao, Xin-Ming Su, Qin Chinese Acad Med Sci Inst Canc Dept Pathol Beijing 100021 Peoples R China Chinese Acad Med Sci Canc Hosp Dept Pathol Beijing 100021 Peoples R China Peking Union Med Coll Beijing 100021 Peoples R China Inner Mongolia Med Coll Affiliated Hosp 1 Dept Pathol Hohhot 010059 Inner Mongolia Peoples R China Inner Mongolia Med Coll Renmin Hosp Dept Pathol Hohhot 010059 Inner Mongolia Peoples R China Chinese Acad Med Sci Inst Canc Dept Radiol Beijing 100021 Peoples R China Chinese Acad Med Sci Canc Hosp Dept Radiol Beijing 100021 Peoples R China

AIM: To establish a model for prognosis assessment of extranodal follicular dendritic cell (FDC) ***: Nine lesions were examined by routine and molecular *** factors from the new cases and 97 reported cases were analyzed for their prognostic ***: The current lesions were found in f ive male and four female patients,located mainly in the head and neck area and averaging 7.2 cm in *** patients had recurrence or metastasis and three remained free of *** 106 patients (male/female ratio,1.1:1) were aged from 9 to 82 years (median,44 years).The tumor sizes ranged from 1.5 to 21 cm (mean,7.4 cm).Abdominal/pelvic region was affected most frequently (43%).Surgical resection was performed in 100 patients,followed by radiation and/or chemotherapy in 35 of ***-up data were available in 91 cases,covering a period of 3-324 mo (mean,27 mo;median,19 mo).Of the informative cases,38 (42%) had recurrence or metastasis,and 12 (13%) died of the *** tumors were classif ied histologically into lowand high-grade lesions.A size ≥ 5 cm (P = 0.003),highgrade histology (P = 0.046) and a mitotic count ≥ 5/10 HPF (P = 0.013) were associated with tumor *** lesions were def ined as low-,intermediateand high-risk tumors,and their recurrence rates were 16%,46% and 73%,and their mortality rates 0%,4% and 45%,***: Extranodal FDC tumors behave like soft tissue *** clinical outcomes are variable and can be evaluated according to their sizes and grades.

关键词： Extranodal follicular dendritic cell sarcoma Prognosis assessment Histologic grade Immunohistochemistry in situ hybridization Mutation detection

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LINE-1 family member GCRG123 gene is up-regulated in human gastric signet-ring cell carcinoma

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World Journal of Gastroenterology 2008年第5期14卷 758-763页

作者： Gang-Shi Wang Meng-Wei Wang Ben-Yan Wu Xin-Yan Yang Wei-Hua Wang Wei-Di You Department of Gerontal Gastroenterology General Hospital of Chinese PLA Beijing 100853 China

AIM： To analyze the expression profiles of a human gastriccancer-related gene, GCRG123, in human gastric signetring cell carcinoma tissues, and to perform bioinformatics analysis on GCRG123.METHODS： in situ hybridization was used to explore the GCRG123 expression pattern in paraffin-embedded gastric tissues, including 15 cases of signet-ring cell carcinoma, 15 of intestinal-type adenocarcinoma, and 15 of normal gastric mucosa. Northern blotting was used to analyze the differences in GCRG123 expression between stomach signet-ring cell carcinoma and intestinal-type adenocarcinoma tissues. Online software, including BLAST, Multalin and BLAT, were applied for bioinformatics analysis. National Center for Biotechnology Information （NCBI） and the University of California Santa Cruz （UCSC） databases were used for the analyses.RESULTS： The in situ hybridization signal appeared as blue precipitates restricted to the cytoplasm. Ten out of 15 cases of gastric signet ring cell carcinoma, normal gastric mucosal epithelium and pyloric glands showed high GCRG123 expression. Low GCRG123 expression was observed in gastric intestinal-type adenocarcinoma and normal gastric glands. Northern blotting revealed that GCRG123 was up-regulated in signet-ring cell carcinoma tissue but down-regulated in intestinal-type adenocarcinoma tissue. BLAST and Multalin analyses revealed that the GCRG123 sequence had 92% similarity with the ORF2 sequence of human long interspersed nuclear element retrotransposons （LINE-1, L1）. BLAT analysis indicated that GCRG123 mapped to all chromosomes. GCRG123 was found to integrate in the intron-17 and -23 of Rb, 5＇ flanking region of IL-2 and clotting factor Ⅸ genes.CONCLUSION： GCRG123, an active member of the Lt family, was up-regulated in human gastric signet-ring cell carcinoma.

关键词： Stomach Signet-ring cell carcinoma GCRG123 LINE-1 in situ hybridization Bioinformatics

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Lonicera japonica-induced inhibition of interleukin-1 beta thermogenesis and E-type prostaglandin receptor-3 expression in the preoptic area of rabbits

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Neural Regeneration Research 2008年第2期3卷 204-207页

作者： Jun Dong Rongbo Tu Rui Pan Xinhua Xie Department of Pathophysiology Medical College of Jinan University Guangzhou 510632 Guangdong Province China Department of Orthopaedics The First Affiliated Hospital of Jinan University Guangzhou 510632 Guangdong Province China

BACKGROUND： It has been shown that interleukin-1β（IL-1β） can induce fever by activating vascular endothelial cells and macrophages of the supraoptic crest to generate prostaglandin E2, which binds with receptors of the thermo-sensitive hypothalamic neurons. Lonicera japonica is one of the medicinal plants used widely in Asia for its antipyretic properties. However, these mechanisms have not yet been intensively studied. OBJECTIVE： To investigate the antipyretic effect and mechanisms of Lonicera japonica on IL-1β- induced febrile New Zealand rabbits by observing expression changes of E-type prostaglandin receptor-3 （EP3） mRNA in the preoptic anterior hypothalamus （POAH）. DESIGN： A randomized controlled study. SETTING： Electrophysiological Laboratory at the Department of Pathophysiology, Medical College of Jinan University; Department of Orthopaedics, First Hospital Affiliated to Medical College of Jinan University. MATERIALS： The experiment was performed from April to December 2005, using a total of 32 New Zealand white rabbits of both sexes, weighing 1.5 2.0 kg. All the animal experiments were performed according to the internationally accepted ethical guidelines. Lonicera japonica injection was purchased from Huanghe pharmaceutical factory of Xi＇an, China. IL-1βwas purchased from Sigma, USA. METHODS： A total of 32 rabbits were divided randomly into four groups： ① Normal saline （NS） control group;② Lonicerajaponica treatment group; ③ IL-1βtreatment group; and ④Lonicerajaponica plus IL-1βtreatment group. In the first 3 groups, the rabbits were given separate intravenous （i.v.） injections of l mL NS, l mL Lonicera japonica, and 100 ng IL-l β （dissolved in 0.9% NS without pyrogen）. In the Lonicerajaponica plus IL-1βgroup each rabbit was given i.v. injections of l mL NS and, 30 minutes later, 100 ng IL-1 β. MAIN OUTCOME MEASURES： Colonic temperature of each rabbit was measured at 0, 10, 20, 30, 40, 50, 60, and 70 minutes after injection and the maximum temperat

关键词： Lonicera japonica interleukin-1β antipyretic preoptic anterior hypothalamus E-type prostaglandin receptor in situ hybridization

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Expression of somatostatin mRNA in various differentiated types of gastric carcinoma

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World Journal of Gastroenterology 1998年第1期4卷 53-56页

作者： ZHANG Qin-Xian DOU Ying-Li SHI Xue-Yi DING Yi Dept. of Histology and Embryology Henan Medical University Zhengzhou 450052 Henan Province China Henan Medical University China

IM METHODS By using in situ hybridization and immunohistochemical techniques, the expression of somatostatin mRNA and somatostatin immunoreactivity in the normal gastric mucosa, the poorly, moderately and welldifferentiated gastric carcinomas, and various clinical stages of carcinoma were observed.RESULTS In comparison with the normal gastric mucosa, the significantly increased expression of somatostatin mRNA positive cells was displayed in gastric carcinoma (t=2681, Phybridization signals in stages Ⅲ and Ⅳ of gastric carcinoma were significantly higher than those in stages Ⅰ and Ⅱ. The results of somatostatin immunoreactivity were consistent with those of in situ hybridization.CONCLUSION The alteration of the expression of somatostatin mRNA was associated with the development of gastric carcinoma and may play an important role in the process of tumor differentiation..

关键词： stomach neoplasms somatostatin RNA,messenger in situ hybridization immunohistochemistry

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Oligodendrocyte transcription factor 1 mRNA and protein expression in organotypic rat brain slices

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Neural Regeneration Research 2010年第21期5卷 1639-1643页

作者： Hong Cui Lijun Yang Dezhuang Huang Wandong Zhang Weijuan Han Yanqing Yao Wenxing Jiang Pediatrics Department Beijing Friendship Hospital Capital Medical University Beijing 100050 China Hepatitis Institute Beijing YouAn Hospital Capital Medical University Beijing 100054 China Institute for Biological Sciences National Research Council of Canada Ottawa K1A OR6 Ontario Canada

Numerous studies have confirmed that oligodendrocyte transcription factor 1 （Olig-1） is vital for myelin repair. However, the effects of hypoxia and ischemia on Olig-1 expression remain unknown. In this study, Olig-1 mRNA and protein expressions were analyzed by in situ hybridization and immunohistochemistry, to determine the expression profile of Olig-1 in rat brain slices exposed to hypoxia and ischemia. Brains were obtained from 2-day-old Sprague-Dawley rats, and sections were randomly assigned to control and hypoxia/ischemia groups. Hematoxylin-eosin staining revealed karyorrhexis and karyopyknosis in cells from the hypoxia/ischemia group. Under electron microscopy, mitochondria swelling and neuropil edema were observed in the hypoxiaJischemia group. Olig-1 mRNA and protein expressions were increased at 1 day after hypoxia and ischemia treatment. These results suggest that in situ hybridization and immunohistochemistry could be used simultaneously to detect mRNA and protein expression in brain slices.

关键词： oligodendrocyte transcription factor 1 double staining brain slices immunohistochemistry in situ hybridization hypoxia and ischemia

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Evidence of human papilloma virus infection and its epidemiology in esophageal squamous cell carcinoma

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World Journal of Gastroenterology 2006年第9期12卷 1352-1355页

作者： Pin-Fang Yao Guang-Can Li Jin Li He-Shun Xia Xiao-Ling Yang Huan-Yuan Huang You-Gao Fu Rui-Qin Wang Xi-Yin Wang Ju-Wei Sha Hubei Cancer Institute Wuhan 430079 Hubei ProvinceChina College of Life Science South-center University for Nationlities Wuhan 430074 Hubei Province China Department of Pathology Hubei Cancer Hospital Wuhan 430079 Hubei Provence China Zhongxiang Chaihu Hospital Zhongxiang 431900 Hubci Province China

AIM： To look for the evidence of human papilloma virus （HPV） infection in esophageal squamous cell carcinomas （ESCC） and to investigate the potential role and epidemiology of HPV infection in the pathogenesis of esophageal carcinomas in Henan emigrants. METHODS： Papilloma virus （PV） and HPV were determined by UltrasensiveTM S-P immunohistochemistry （IHC） and in situ hybridization （ISH） in esophageal carcinoma tissues （82 cases） and the normal mucosa （40 cases）. RESULTS： IHC revealed that the positive rate of PV was 75.0%, 68.18% and 72.5% respectively while the HPV （16/18-E6） positive rate was 45.0%, 36.36%, 37.5%, respectively in esophageal carcinoma tissue specimens from Henan emigrants,the local citizens and patients in Hubei Cancer Hospital. The PV and HPV （16/18-E6） were negative in all normal esophageal mucosa specimens. No correlation was found between HPV in esophageal squamous cell carcinoma tissues and in grade 1-3 esophageal squamous cell carcinoma cells. in situ hybridization showed that the HPV （16/18） DNA positive rate was 30.0%, 31.8%, 25.0%, respectively in the 3 groups of samples. No positive hybridization signal was found in 40 normal esophageal mucosa specimens. The positive rate of HPV （16/18） DNA in the esophageal carcinoma specimens was significantly higher than that in normal mucosa specimens （P〈0.05）. The positive rate was not different among the 3 groups of esophageal carcinoma tissue specimens （P〉0.05）.CONCLUSION： HPV infection is high in esophageal carcinoma of Henan emigrants, local residents and patients in Hubei Cancer Hospital. HPV is closely related with esophageal squarnous cell carcinoma. HPV infection may play an important role in esophageal squarnous cell carcinoma.

关键词： Human papillomavirus Esophageal squamous cell carcinoma Immunohistochemistry in situ hybridization

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