Expression pattern and level of ING5 protein in normal and cancer tissues.

Inhibitor of growth family 5 (ING5) functions as a type-II tumor suppressor gene and exerts an important role in DNA repair, apoptotic induction, proliferative inhibition, chromatin remodeling and the invasion process. In the present study, immunohistochemistry was performed to characterize the expression profile of ING5 protein on a tissue microarray containing mouse and human normal tissues, and human cancer tissues, including hepatocellular (n=62), renal clear cell (n=62), pancreatic (n=62), esophageal squamous cell (n=45), cervical squamous cell (n=31), breast (n=144), gastric (n=196), colorectal (n=96), endometrial (n=96) and lung carcinoma (n=192). In the mouse tissues, ING5 expression was detected in the cytoplasm of neurons, the nephric tubule and glomerulus, alveolar epithelium, gastrointestinal glands, squamous epithelium of the skin and skeletal muscles. By contrast, ING5 was localized to the cell nucleus in breast tissues. In human tissues, ING5 protein was primarily localized in the cytoplasm. However, ING5 was detected in the cytoplasm and nucleus in various types of normal tissues, including the tongue, stomach, intestine, lung and breast. In total, ING5 expression was detected in 400/986 cancer tissues (40.6%). In the majority of cases, ING5 expression was observed to be restricted to the cytoplasm. However, ING5 was also detected in the nucleus in a number of cancer tissues, including gastric, colorectal and lung carcinoma. Notably, ING5 was more frequently expressed in breast (79.9%), colorectal (56.3%) and endometrial carcinoma (50.0%). The incidence of ING5 expression in hepatocellular carcinoma (14.5%) and pancreatic carcinoma (22.6%) was low. These findings indicate that ING5 may be involved in cell regeneration and be associated with colorectal carcinogenesis.

[Effects of conservation tillage on soil water condition and winter wheat yield in farmland]. / 保护性耕作对农田土壤水分和冬小麦产量的影响.

Conservation tillage is one of the most important agricultural management measures on soil water conservation and crop yield increments. Based on long-term experiment during 2011 to 2016, effects of different tillage treatments, including conventional tillage, no-tillage and subsoiling treatments, on soil water condition, crop yield and water use efficiency were analyzed. The results showed that the average and relative soil water conservation rate was 7.3% and -0.68% at jointing stage of winter wheat under no tillage and subsoiling treatments, respectively. Compared with conventional tillage, soil water storage significantly increased in 0-60 cm soil layer at jointing stage of winter wheat under no-tillage. Besides, the mean values of soil water content significantly increased in 0-100 cm soil layer at jointing, flowering, filling, and harvesting stages under no tillage treatment while that was not significantly increased at jointing stage under subsoiling treatment. Furthmore, no-tillage treatment significantly increased the yield and water use efficiency of winter wheat, especially in the dry years. Therefore, soil moisture conservation ability and yield increment of no tillage was better than that of the subsoiling treatment in dry years.

ING5-mediated antineuroblastoma effects of suberoylanilide hydroxamic acid.

Neuroblastoma is the most common extracranial solid neuroendocrine cancer and is one of the leading causes of death in children. To improve clinical outcomes and prognosis, discovering new promising drugs and targeted medicine is essential. We found that applying Suberoylanilide hydroxamic acid (SAHA; Vorinostat, a histone deacetylase inhibitor) and MG132 (a proteasome inhibitor) to SH-SY5Y cells synergistically suppressed proliferation, glucose metabolism, migration, and invasion and induced apoptosis and cell cycle arrest. These effects occurred both concentration and time dependently and were associated with the effects observed with inhibitor of growth 5 (ING5) overexpression. SAHA and MG132 treatment increased the expression levels of ING5, PTEN, p53, Caspase-3, Bax, p21, and p27 but decreased the expression levels of 14-3-3, MMP-2, MMP-9, ADFP, Nanog, c-myc, CyclinD1, CyclinB1, and Cdc25c concentration dependently, similar to ING5. SAHA may downregulate miR-543 and miR-196-b expression to enhance the translation of ING5 protein, which promotes acetylation of histones H3 and H4. All three proteins (ING5 and acetylated histones H3 and H4) were recruited to the promoters of c-myc, Nanog, CyclinD1, p21, and p27 for complex formation, thereby regulating the mRNA expression of downstream genes. ING5 overexpression and SAHA and/or MG132 administration inhibited tumor growth in SH-SY5Y cells by suppressing proliferation and inducing apoptosis. The expression of acetylated histones H3 and ING5 may be closely linked to the tumor size of neuroblastomas. In summary, SAHA and/or MG132 can synergistically suppress the malignant phenotypes of neuroblastoma cells through the miRNA-ING5-histone acetylation axis and via proteasomal degradation, respectively. Therefore, the two drugs may serve as potential treatments for neuroblastoma.

The roles of ING5 in gliomas: a good marker for tumorigenesis and a potential target for gene therapy.

To elucidate the anti-tumor effects and molecular mechanisms of ING5 on glioma cells, we overexpressed it in U87 cells, and examined the phenotypes and their relevant molecules. It was found that ING5 overexpression suppressed proliferation, energy metabolism, migration, invasion, and induced G2/M arrest, apoptosis, dedifferentiation, senescence, mesenchymal- epithelial transition and chemoresistance to cisplatin, MG132, paclitaxel and SAHA in U87 cells. There appeared a lower expression of N-cadherin, Twist, Slug, Zeb1, Zeb2, Snail, Ac-H3, Ac-H4, Cdc2, Cdk4 and XIAP, but a higher expression of Claudin 1, Histones 3 and 4, p21, p53, Bax, ß-catenin, PI3K, Akt, and p-Akt in ING5 transfectants. ING5 overexpression suppressed tumor growth of U87 cells in nude mice by inhibiting proliferation and inducing apoptosis. Down-regulated ING5 expression was closely linked to the tumorigenesis and histogenesis of glioma. These data indicated that ING5 expression might be considered as a good marker for the tumorigenesis and histogenesis of gliomas. It might be employed as a potential target for gene therapy of glioma. PI3K/Akt or ß-catenin/TCF-4 activation might be positively linked to chemotherapeutic resistance, mediated by ING5.

Cytokeratin 19 promoter directs the expression of Cre recombinase in various epithelia of transgenic mice.

Cytokeratin 19 (K19) is expressed in various differentiated cells, including gastric, intestinal and bronchial epithelial cells, and liver duct cells. Here, we generated a transgenic mouse line, K19-Cre, in which the expression of Cre recombinase was controlled by the promoter of K19. To test the tissue distribution and excision activity of Cre recombinase, K19-Cre transgenic mice were bred with Rosa26 reporter strain and a mouse strain that carries PTEN conditional alleles (PTENLoxp/Loxp). At mRNA level, Cre was strongly expressed in the stomach, lung and intestine, while in stomach, lung, and liver at protein level. The immunoreactivity to Cre was strongly observed the cytoplasm of gastric, bronchial and intestinal epithelial cells. Cre activity was detectable in gastric, bronchial and intestinal epithelial cells, according to LacZ staining. In K19-Cre/PTEN Loxp/Loxp mice, PTEN was abrogated in stomach, intestine, lung, liver and breast, the former two of which were verified by in situ PCR. There appeared breast cancer with PTEN loss. These data suggest that K19 promoter may be a useful tool to study the pathophysiological functions of cytokeratin 19-positive cells, especially gastrointestinal epithelial cells. Cell specificity of neoplasia is not completely attributable to the cell-specific expression of oncogenes and cell-specific loss of tumor suppressor genes.

The suppressing effects of BTG3 expression on aggressive behaviors and phenotypes of colorectal cancer: An in vitro and vivo study.

Here, we found that down-regulated expression of BTG3 might be positively correlated with colorectal carcinogenesis and its overexpression suppressed proliferation, glycolysis, mitochondrial respiration, cell cycle progression, migration, and invasion, and induced apoptosis, senescence and differentiation in SW480 and SW620 cells. After treated with cisplatin, MG132, paclitaxel and SAHA, BTG3 transfectants exhibited lower viability and higher apoptosis than the control in both time- and dose-dependent manners. BTG3 overexpression up- regulated the protein expression of Cyclin E, p16, p27, NF-κB, p38α/ß, XIAP, Bcl-2, ATG14 and p53, but down-regulated the mRNA expression of MRP1, BCRP, and mTOR in SW480 and SW620 cells. BTG3 overexpression inhibited tumor growth of SW620 cells by suppressing proliferation and inducing apoptosis. It was suggested that down-regulated BTG3 expression might be considered as a marker for colorectal carcinogenesis. BTG3 overexpression might reverse the aggressive phenotypes and be employed as a potential target for gene therapy of colorectal cancer.

The in vitro and vivo effects of nuclear and cytosolic parafibromin expression on the aggressive phenotypes of colorectal cancer cells: a search of potential gene therapy target.

Down-regulated parafibromin is positively linked to the pathogenesis of parathyroid, lung, breast, ovarian, gastric and colorectal cancers. Here, we found that wild-type (WT) parafibromin overexpression suppressed proliferation, tumor growth, induced cell cycle arrest and apoptosis in colorectal cancer cells (p<0.05), but it was the converse for mutant-type (MT, mutation in nucleus localization sequence) parafibromin (p<0.05). Both WT and MT transfectants inhibited migration and invasion, and caused better differentiation (p<0.05) of cancer cells. WT parafibromin transfectants showed the overexpression of Cyclin B1, Cyclin D1, Cyclin E, p38, p53, and AIF in HCT-15 and HCT-116 cells, while MT parafibromin only up-regulated p38 expression. There was lower mRNA expression of bcl-2 in parafibromin transfectants than the control and mock, while higher expression of c-myc, Cyclin D1, mTOR, and Raptor. According to transcriptomic analysis, WT parafibromin suppressed PI3K-Akt and FoxO signaling pathways, while MT one promoted PI3K-Akt pathway, focal adhesion, and regulation of actin cytoskeleton. Parafibromin was less expressed in colorectal cancer than paired mucosa (p<0.05), and inversely correlated with its differentiation at both mRNA and protein levels (p<0.05). These findings indicated that WT parafibromin might reverse the aggressive phenotypes of colorectal cancer cells and be employed as a target for gene therapy. Down-regulated parafibromin expression might be closely linked to colorectal carcinogenesis and cancer differentiation.

The pathological behaviors and prognostic factors of Chinese and Japanese colorectal cancers from general hospitals: a comparative study of the inpatients with surgical operation.

Here, we collected the information of 17304 and 2014 inpatients with colorectal cancer (CRC) from general hospitals of China and Japan respectively, and analyzed microscopic and macroscopic aspects, even stratified by the age and gender. It was found that Chinese CRC patients showed younger prone, more rectal and ascending cancers, less sigmoid and transverse cancers, larger size, less invasion into lymphatic system or metastasis into lymph node, and poorer differentiation than Japanese ones (p < 0.05). TNM staging was employed as an independent factor for the prognosis of the CRC patients regardless of the country (p < 0.05). Female patients showed larger tumor size, easier invasion and metastasis into lymphatic system, and worse differentiation than males (p < 0.05). The younger patients displayed frequent invasion and metastasis into lymphatic system, and poor differentiation in comparison to elder ones (p < 0.05). These findings demonstrated that Japanese patients seemed to have more invasion and metastasis due to standard and precise operation and pathological diagnosis accuracy. Actually, Chinese patients had more aggressive pathological characteristics and a poorer prognosis. Therefore, it is essential to establish a routine screening methodology, a standard treatment system and postoperative diagnosis protocol for the prevention and therapeutics of Chinese CRC patients, especially for female and young patients.

The down-regulated ING5 expression in lung cancer: a potential target of gene therapy.

ING5 can interact with p53, thereby inhibiting cell growth and inducing apoptosis. We found that ING5 overexpression not only inhibited proliferation, migration, and invasion, but also induced G2 arrest, differentiation, autophagy, apoptosis, glycolysis and mitochondrial respiration in lung cancer cells. ING5 transfection up-regulated the expression of Cdc2, ATG13, ATG14, Beclin-1, LC-3B, AIF, cytochrome c, Akt1/2/3, ADFP, PFK-1 and PDPc, while down-regulated the expression of Bcl-2, XIAP, survivin,ß-catenin and HXK1. ING5 transfection desensitized cells to the chemotherapy of MG132, paclitaxel, and SAHA, which paralleled with apoptotic alteration. ING5 overexpression suppressed the xenograft tumor growth by inhibiting proliferation and inducing apoptosis. ING5 expression level was significantly higher in normal tissue than that in lung cancer at both protein and mRNA levels. Nuclear ING5 expression was positively correlated with ki-67 expression and cytoplasmic ING5 expression. Cytoplasmic ING5 expression was positively associated with lymph node metastasis, and negatively with age, lymphatic invasion or CPP32 expression. ING5 expression was different in histological classification: squamous cell carcinoma > adenocarcinoma > large cell carcinoma > small cell carcinoma. Taken together, our data suggested that ING5 downregulation might involved in carcinogenesis, growth, and invasion of lung cancer and could be considered as a promising marker to gauge the aggressiveness of lung cancer. It might be employed as a potential target for gene therapy of lung cancer.

The roles of parafibromin expression in ovarian epithelial carcinomas: a marker for differentiation and prognosis and a target for gene therapy.

Parafibromin is a protein encoded by hyperparathyroidism 2 (HRPT2) and its downregulated expression is involved in the pathogenesis of parathyroid, breast, gastric, colorectal, lung, head and neck cancers. We aimed to investigate the roles of parafibromin expression in tumorigenesis, progression, or prognostic evaluation of ovarian cancers. HRPT2-expressing plasmid was transfected into ovarian cancer cells with the phenotypes and related molecules examined. The messenger RNA (mRNA) and protein expression of parafibromin were also examined in ovarian normal tissue, benign and borderline tumors and cancers by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, or immunohistochemistry respectively. It was found that parafibromin overexpression caused a lower growth, migration and invasion, higher sensitivity to cisplatin and apoptosis than the mock and control (P < 0.05). The transfectants showed the hypoexpression of phosphoinositide 3-kinase (PI3K), Akt, p70 ribosomal protein S6 kinase (p70s6k), Wnt5a, B cell lymphoma-extra large (Bcl-xL), survivin, vascular endothelial growth factor (VEGF) and matrix metallopeptidase 9 (MMP-9) than the mock and control at both mRNA and protein levels (P < 0.05). According to real-time PCR, parafibromin mRNA level was lower in ovarian benign tumors and cancers than normal ovary (P < 0.05), while parafibromin was strongly expressed in metastatic cancers in omentum than primary cancers by Western blot. Immunohistochemically, parafibromin expression was stronger in primary cancers than that in ovarian normal tissue (P < 0.05) but weaker than the metastatic cancers (P < 0.05) with a positive correlation with dedifferentiation, ki-67 expression and the lower cumulative survival rate (P < 0.05). These findings indicate that parafibromin downregulation might promote the pathogenesis, dedifferentiation and metastasis of ovarian cancers possibly by suppressing aggressive phenotypes, such as proliferation, cell cycle, apoptosis, migration and invasion.

Nickel-catalyzed Kumada reaction of tosylalkanes with Grignard reagents to produce alkenes and modified arylketones.

Open a new door: The first example of alkene synthesis from alkyl electrophiles with Grignard reagents using the Kumada cross-coupling reaction strategy is reported. This method opens a new door for the Kumada cross-coupling reaction, allowing alkenes to be prepared from the reaction of tosylalkanes with Grignard reagents.

Copper-catalyzed C-H oxidation/cross-coupling of α-amino carbonyl compounds.

Keeping options open: the new and mild title reaction involving indoles selectively furnishes 1 and 2 with the aid of tert-butyl hydroperoxide (TBHP). The method represents the first example of a copper-catalyzed α arylation of α-amino carbonyl substrates leading to α-aryl α-imino and α-aryl α-oxo carbonyl compounds using a C-H oxidation strategy.

[Responses of winter wheat photosynthetic characteristics and chlorophyll content to water-retaining agent and N fertilizer].

The effects of water-retaining agent (60 kg x hm(-2)) and nitrogen fertilizer (0, 225, and 450 kg x hm(-2)) on the leaf photosynthetic characteristics, chlorophyll content, and water utilization of winter wheat at jointing and grain-filling stages were studied under field conditions. In all treatments, the net photosynthetic rate, stomata conductance, intercellular CO2 concentration, water use efficiency, and chlorophyll content were greater at grain-filling stage than at jointing stage. Under nitrogen fertilization but without water-retaining agent application, the water use efficiency (WUE) of single leaf at jointing stage increased with increasing nitrogen fertilization rate, while the net photosynthetic rate, stomata conductance, intercellular CO2 concentration, and transpiration rate decreased after an initial increase. The chlorophyll content was the highest under 225 kg x hm(-2) nitrogen fertilization. In the treatments of water-retaining agent application, the intercellular CO2 con- centration decreased with increasing nitrogen application rate, but the net photosynthetic rate, transpiration rate, and WUE increased. The application of water-retaining agent or its combination with nitrogen fertilization increased the chlorophyll content, but excessive nitrogen fertilization had lesser effects. At grain-filling stage, applying nitrogen fertilizer alone significantly increased the net photosynthetic rate and WUE, but decreased the stomata conductance, intercellular CO2 concentration, and transpiration rate. The chlorophyll content increased with increasing nitrogen application rate. After applying water-retaining agent and with the increase of nitrogen fertilization rate, the photosynthetic rate and WUE decreased after an initial increase, while the intercellular CO2 concentration and transpiration rate were in adverse but still lower than those without water-retaining agent application. The stomata conductance increased with increasing nitrogen fertilization rate. The chlorophyll content increased significantly under the application of water-retaining agent, but somewhat decreased under the combined application of water-retaining agent and nitrogen fertilizer. The application of both water-retaining agent and nitrogen fertilizer increased the 1000 grain mass, grain yield, and water production efficiency of winter wheat significantly, with the best effect in the treatment of water-retaining agent with 225 kg x hm(-2) nitrogen fertilization.

[Effects of ground cover and water-retaining agent on winter wheat growth and precipitation utilization].

An investigation was made at a hilly upland in western Henan Province to understand the effects of water-retaining agent (0, 45, and 60 kg x hm(-2)), straw mulching (3000 and 6000 kg x hm(-2)), and plastic mulching (thickness < 0.005 mm) on winter wheat growth, soil moisture and nutrition conditions, and precipitation use. All the three measures promoted winter wheat growth, enhanced grain yield and precipitation use efficiency, and improved soil moisture and nutritional regimes. These positive effects were more obvious when the straw- or plastic mulching was combined with the use of water-retaining agent. Comparing with the control, all the measures increased the soil moisture content at different growth stages by 0.1%-6.5%. Plastic film mulching had the best water-retention effect before jointing stage, whereas water-retaining agent showed its best effect after jointing stage. Soil moisture content was the lowest at flowering and grain-filling stages. Land cover increased the grain yield by 2.6%-20.1%. The yield increment was the greatest (14.2%-20.1%) by the combined use of straw mulching and water-retaining agent, followed by plastic mulching combined with water-retaining agent (11.9% on average). Land cover also improved the precipitation use efficiency (0.4-3.2 kg x mm(-1) x hm(-2)) in a similar trend as the grain yield. This study showed that land cover and water-retaining agent improved soil moisture and nutrition conditions and precipitation utilization, which in turn, promoted the tillering of winter wheat, and increased the grain number per ear and the 1000-grain mass.

[Effects of different application rates of water-retaining agent on root physiological characteristics of winter wheat at its different growth stages].

A field experiment was conducted at the Yuzhou Experimental Base of Henan Province to study the effects of different application rates (0, 30, 60, and 90 kg x hm(-2)) of water-retaining agent (WRA) on the root physiological characteristics, biomass, and grain yield of two winter wheat cultivars Zhengmai-9694 and Aikang-58, aimed to probe into the action mechanisms of WRA on the root system of winter wheat at its different growth stages. The application of WRA decreased the root membrane permeability and soluble sugar content, and increased the root vigor. After the application of WRA, the Zhengmai-9694 at its different growth stages had a greater decrement of root membrane permeability, compared with Aikang-58. In all treatments except 90 kg x hm(-2) of WRA, the root vigor of Aikang-58 was obviously higher than that of Zhengmai-9694. At booting and grain-filling stages, the root soluble sugar content of Zhengmai-9694 decreased much more than that of Aikang-58. In the whole growth period of the two cultivars, their root membrane permeability and soluble sugar content were the lowest in treatment 60 kg x hm(-2) of WRA, and no significant differences were observed between treatments 60 and 90 kg x hm(-2) of WRA. The root vigor of Zhengmai-9694 increased remarkably with the increasing rate of WRA application, while that of Aikang-58 was the highest in treatment 60 kg x hm(-2) of WRA. The application of WRA also increased root biomass, and at jointing and booting stages, the root biomass of Aikang-58 was much higher than that of Zhengmai-9694. However, at grain-filling stage, the biomass of Aikang-58 in treatments 60 and 90 kg x hm(-2) of WRA was lower than that of Zhengmai-9694. Treatment 60 kg x hm(-2) of WRA had the highest grain yield of the two cultivars. It was concluded that WRA had more significant effects on Zhengmai-9694 than on Aikang-58, and applying 60 kg x hm(-2) of WRA could obtain the best effect.