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Dose dependent effects of TPCA-1 on STAT3 activation.

TPCA-1 inhibits endogenous STAT3 activity with or without exogenous IL6 in NSCLC with EGFR mutation.

Abundance of IL6 and COX2 in NSCLC cells and TPCA-1 inhibits their transcription.

TPCA-1 blocks cell cycle in G2/M in HCC827 cells.

Epidermal growth factor receptor (EGFR) is a clinical therapeutic target to treat a subset of non–small cell lung cancer (NSCLC) harboring EGFR mutants. However, some patients with a similar kind of EGFR mutation show intrinsic resistance to tyrosine kinase inhibitors (TKI). It indicates that other key molecules are involved in the survival of these cancer cells. We showed here that 2-[(aminocarbonyl)amino]-5 -(4-fluorophenyl)-3- thiophenecarboxamide (TPCA-1), a previously reported inhibitor of IκB kinases (IKK), blocked STAT3 recruitment to upstream kinases by docking into SH2 domain of STAT3 and attenuated STAT3 activity induced by cytokines and cytoplasmic tyrosine kinases. TPCA-1 is an effective inhibitor of STAT3 phosphorylation, DNA binding, and transactivation in vivo. It selectively repressed proliferation of NSCLC cells with constitutive STAT3 activation. In addition, using pharmacologic and genetic approaches, we found that both NF-κB and STAT3 could regulate the transcripts of interleukin (IL)-6 and COX-2 in NSCLC harboring EGFR mutations. Moreover, gefitinib treatment only did not efficiently suppress NF-κB and STAT3 activity. In contrast, we found that treatment with TKIs increased phosho-STAT3 level in target cells. Inhibiting EGFR, STAT3, and NF-κB by combination of TKIs with TPCA-1 showed increased sensitivity and enhanced apoptosis induced by gefitinib. Collectively, in this work, we identified TPCA-1 as a direct dual inhibitor for both IKKs and STAT3, whereas treatment targeting EGFR only could not sufficiently repress NF-κB and STAT3 pathways for lung cancers harboring mutant EGFR. Therefore, synergistic treatment of TPCA-1 with TKIs has potential to be a more effective strategy for cancers. Mol Cancer Ther; 13(3); 617–29. ©2014 AACR.

EGFR silence inhibits STAT3 activation in MDA-MB-231 cells.

Supplementary method and figure legends

TPCA-1 inhibits STAT3 phosphorylation.

Combination of TPCA-1 increases sensitivity to gefitinib in TKI insensitive NSCLC.

Significance Tamoxifen is beneficial in treating estrogen receptor–positive breast cancer, but resistance to this treatment eventually ensues. A method to identify mechanisms of tamoxifen resistance identified the histone deacetylase ZIP, leading to the finding that increased expression of the tyrosine kinase JAK2 is one important factor. As a result of this discovery, it may be possible to use an inhibitor of JAK2 to block the aberrant activation of STAT3 caused by ZIP deficiency to help overcome or prevent tamoxifen resistance., Tamoxifen, a widely used modulator of the estrogen receptor (ER), targets ER-positive breast cancer preferentially. We used a powerful validation-based insertion mutagenesis method to find that expression of a dominant-negative, truncated form of the histone deacetylase ZIP led to resistance to tamoxifen. Consistently, increased expression of full-length ZIP gives the opposite phenotype, inhibiting the expression of genes whose products mediate resistance. An important example is JAK2. By binding to two specific sequences in the promoter, ZIP suppresses JAK2 expression. Increased expression and activation of JAK2 when ZIP is inhibited lead to increased STAT3 phosphorylation and increased resistance to tamoxifen, both in cell culture experiments and in a mouse xenograft model. Furthermore, data from human tumors are consistent with the conclusion that decreased expression of ZIP leads to resistance to tamoxifen in ER-positive breast cancer.

N6‐Methyladenosine (m6A) is considered the most common and endogenous modification of eukaryotic RNAs. Highly conserved in many species, m6A regulates RNA metabolism, cell differentiation, cell circadian rhythm, and cell cycle; it also responds to endogenous and exogenous stimuli and is associated with the development of tumors. The m6A methyltransferase complex (MTC) regulates the m6A modification of transcripts and involves two components, methyltransferase‐like enzyme 3 (METTL3) and methyltransferase‐like enzyme 14 (METTL14), and other auxiliary regulatory distinct components. Though with no catalytic effect, METTL14 serves as an RNA‐binding scaffold in MTC, promotes RNA substrate recognition, activates, and escalates the catalytic capability of METTL3, thus accounting for a pivotal member of the complex. It was reported that METTL14 regulates tumor proliferation, metastasis, and self‐renewal, and plays a part in tumorigenesis, tumor progression, and other processes. The present work is a review of the role of METTL14 both as a tumor suppressor and a tumor promoter in the oncogenesis and progression of various tumors, as well as the potential molecular mechanisms., Roles of METTL14 in oncogenesis and development. The abnormal expression of METTL14, an m6A modification editor, affects tumor cell proliferation, metastasis, tumor stem cell self‐renewal, and chemotherapy resistance.

Background Ovarian cancer is the leading cause of death in patients with gynecologic cancer, and circular RNAs (circRNAs) are involved in cancer progression. However, there are limited studies on the roles of circRNAs in ovarian cancer. Methods We designed divergent and convergent primers, used sanger sequencing and RNase R digestion to verify the source of circCRIM1. We detected the expression of circCRIM1 and its parental gene cysteine rich transmembrane BMP regulator 1 (CRIM1) in ovarian cancer and normal ovarian samples via qRT-PCR. MTT viability assay, apoptosis assay, wound healing assay and invasion assay were used to investigate the function of circCRIM1 and CRIM1 in ovarian cancer cell lines OVCAR3 and CAOV3. Mice xenografts experiment was performed. Bioinformatics predicted the microRNAs that bond with circCRIM1 and CRIM1, and dual luciferase reporter system confirmed it. Rescue experiments of microRNAs mimics transfection on the basis of circCRIM1 over-expression were carried out to uncover the mechanism by which circCRIM1 played cancer-promoting roles in ovarian cancer. Results CircCRIM1 was derived from CRIM1 by back-splicing. CircCRIM1 and CRIM1 had higher expression in ovarian cancer than in normal ovarian tissues, and both of them promoted ovarian cancer progression in vitro. In vivo circCRIM1 promoted the growth of tumors. CircCRIM1 and CRIM1 had a positive correlation relationship in the same cohort of ovarian cancer tissues. Bioinformatics predicted and dual luciferase assay confirmed circCRIM1 and CRIM1 bond with miR-145-5p, and circCRIM1 bond with miR-383-5p additionally. CircCRIM1 positively affected the expression of CRIM1. After circCRIM1 was over-expressed, miR-145-5p mimics transfection reversed the expression of CRIM1. Western blot discovered circCRIM1 positively affected the expression of zinc finger E-box binding homeobox 2 (ZEB2). Rescue experiments found miR-383-5p mimics reversed ZEB2 expression and the cancer-promoting effects of circCRIM1. Conclusions CircCRIM1 bond with miR-145-5p to work as competing endogenous RNA (ceRNA) of CRIM1, and circCRIM1 bond with miR-383-5p to improve the expression of ZEB2 in ovarian cancer. CircCRIM1 and CRIM1 promoted the ovarian cancer progression and supplied a novel insight into the researches of ovarian cancer.

The Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway plays a vital role in immunity, cell division, cell death and tumor formation. Disrupted JAK-STAT signaling may lead to various diseases, especially cancer and immune disorders. Because of its importance, this signaling pathway has received significant attention from the pharmaceutical and biotechnology industries as a therapeutic target for drug design. However, few JAK or STATs inhibitors have been developed for cancer treatment. We used an in vitro STAT3 luciferase reporter assay to find novel inhibitors that could effectively block the JAK-STAT pathway. In our study, we screened 16,081 drug-like chemicals and found that atopaxar hydrobromide (AHB) is a specific inhibitor of JAK-STAT3 signaling. Our results suggest that AHB not only blocks constitutively activated and cytokine-induced STAT3 phosphorylation but also inhibits JAK1 and JAK2 phosphorylation. Moreover, AHB induces G1 phase cell cycle arrest, which stops cancer cell growth and induces apoptosis. AHB also inhibited tumor cell growth in vivo. In conclusion, AHB is a potential inhibitor that could be developed as a JAK-STAT pathway drug.

Transforming growth factor beta is a key cytokine involved in the pathogenesis of fibrosis in many organs, whereas interleukin-6 plays an important role in the regulation of inflammation. They are both potent angiogenesis inducers with opposite effects on cell survival and apoptosis. TGF-β2 induces apoptosis; in contrast, IL-6 protects cells from apoptosis. The possible interaction between these two cytokines is indicated in various disease states. In this study, we have assessed the effect of TGF-β2 on IL-6 signaling and found that TGF-β2 could strongly inhibit IL-6-induced STAT3 activation and synergy with IL-6 resulting in enhanced SOCS3 expression. Interestingly, IL-6 also slows down the decay of TGF-β2 mRNA. Consistent with this mechanism, we found that TGF-β2 could antagonize IL-6 effect on cell survival in both γ-irradiation and UV light-induced apoptosis. Taken together, the finding shows that TGF-β2 serves as a negative regulator of IL-6 signaling and antagonizes the anti-apoptosis effect of IL-6.

Activation of transcription factor STAT3 is involved in cell proliferation, differentiation, and cell survival. Constitutive activation of STAT3 pathway has been associated with the oncogenesis of various types of cancers. It has been reported that STAT3 plays a key role in the G1 to S phase cell cycle transition induced by the cytokine receptor subunit gp130, through the upregulation of cyclins D1, D2, D3, A, and Cdc25A and the concomitant downregulation of p21 and p27. However, its role in mediating G2-M phase transition has not been studied. The cyclin B1/Cdc2 complex is widely accepted as the trigger of mitosis in all organisms and is believed to be necessary for progression through S phase and keep active during the G2-M transition and progression. In the present study, we found that activation of STAT3 stimulates cyclin B1 and Cdc2 expressions. Deletion and site-directed mutations on cyclin B1 and Cdc2 promoters indicated that E2F element mediates the upregulation of these two promoters in a STAT3-dependent manner. The findings reported here demonstrated that STAT3 participates in modulating G2-M phase checkpoint by regulating gene expressions of cyclin B1 and Cdc2 via E2F.

Yuping Du,1 Ping Gu,2 Yu Cui,2 Yi Wang,2 Juanjuan Ran2 1Department of Neurology, the 904th Hospital of Joint Logistic Support Force, PLA, Wuxi, 214044, People’s Republic of China; 2Department of Neurology, Wuxi No.5 People’s Hospital, Wuxi, 214000, People’s Republic of ChinaCorrespondence: Juanjuan RanDepartment of Neurology, Wuxi No.5 People’s Hospital, Wuxi, Jiangsu Province, People’s Republic of ChinaTel +86-13255109976Fax +86-510-68585555Email 7532164@qq.comPurpose: To develop a nomogram to predict the risk of subsequent vascular events (SVE) at 6-month in Chinese patients with minor ischemic stroke (MIS).Patients and Methods: We performed a retrospective analysis of 260 MIS patients, which were randomly divided into a derivation set (193 cases) and a verification set (67 cases) at a ratio of 3:1. Multi-factor logistic regression was used to construct a predictive model of SVE from the derivation set and verify it in the verification set.Results: Finally, there were 51 cases (19.6%) of SVE in 260 MIS cases. Age, fasting blood glucose, metabolic syndrome, number of lesions found on MRI, and the infarct size were used to construct the prediction model and nomogram. The AUC in the derivation set was 0.901, with a sensitivity of 0.795, a specificity of 0.877, a positive likelihood ratio of 6.443, and a negative likelihood ratio of 0.234. The AUC in the verification set was 0.897, which was not significantly different from the derivation set (P = 0.937). The predictive model based on clinical parameters has good diagnostic efficiency and robustness.Conclusion: The nomogram can provide personalized predictions for the 6-month SVE risk in Chinese MIS patients.Keywords: logistic models, nomograms, brain ischemia, stroke, infarction

Background: Circular RNAs (circRNAs) play important roles in cancer tumorigenesis and progression, and could represent prognostic biomarkers and therapeutic targets. Herein, we demonstrated the role of circNOLC1 in epithelial ovarian cancer (EOC). Methods: CircNOLC1 expression was quantified in ovarian cancer tissues and normal ovarian tissues using quantitative real-time reverse transcriptase. After circNOLC1 overexpression or downregulation using a circNOLC1 overexpression plasmid or circNOLC1 short hairpin RNA (shRNA) transfection, respectively, ovarian cancer cell phenotypes and molecular mechanisms were examined in vivo and in vitro. Results: Circ-NOLC1 expression was higher in EOC tissues than in normal tissues, and was positively associated with FIGO stage, differentiation. Among ovarian cancer cell lines, circ-NOLC1 expression was highest in A2780, and lowest in CAOV3. Overexpression of circ-NOLC1 in CAOV3 cells increased cell proliferation, migration, and invasion, while silencing of circ-NOLC1 in A2780 cells had the opposite effect: however, neither circ-NOLC1 downregulation nor overexpression influenced nucleolar and coiled-body phosphoprotein 1 ( NOLC1 ) mRNA expression. In nude mice with subcutaneous tumors, circ-NOLC1 downregulation decreased tumor growth . Bioinformatic analysis and RNA binding protein immunoprecipitation showed that circNOLC1 could bind to epithelial splicing regulatory protein 1 (ESRP1). In addition, circ-NOLC1 overexpression significantly increased ESRP1 protein levels, and those of ras homolog family member A (RhoA) and cyclin dependent kinase 1 (CDK1); circNOLC1 downregulation had the opposite effects. The tumor-promoting effect of circNOLC1 was inhibited by knockdown of ESRP1 expression in circ-NOLC1 overexpressing cells, which might act by modulating RhoA and CDK1 expression. Conclusion: CircNOLC1 might promote EOC tumorigenesis and development by binding ESRP1 and modulating CDK1 and RhoA expression.

Background. As a newly discovered regulatory RNA, circular RNA (circRNA) has become a hot spot in many tumor pieces of research. In recent years, it has been discovered that circRNAs have multiple biological effects in different stages of cancer. However, the expression pattern and mechanism of circFAT1(e2) in non-small-cell lung cancer (NSCLC) are still unclear. Methods. The expressions of circFAT1(e2) in NSCLC tissues and cell lines were studied. Functionally, CCK-8 and transwell experiments were performed in A549 and H1299. In addition, we also performed a dual-luciferase report analysis to clarify the mechanism of action of circFAT1(e2). Results. circFAT1(e2) was significantly upregulated in NSCLC tissues and cell lines. circFAT1(e2) gene knockdown could significantly inhibit the proliferation, migration, and invasion of NSCLC cells. Loss of function testing found that circFAT1(e2) functioned as an oncogene in NSCLC cells. In addition, circFAT1(e2) acted as a ceRNA to spongy miR-30e-5p, which led to the increase in USP22 and promoted cell growth. Conclusions. The circFAT1(e2)-miR-30e-5p-USP22 axis is a crucial part of the progression of NSCLC. This study suggests that circFAT1(e2) may be an important potential of prognostic prediction and treatment targets for NSCLC patients.

OBJECTIVE: To probe the clinical characteristics of diagnosis and therapy of vascular pulsatile tinnitus(PT) associated with sigmoid sinus-mastoid. METHODS: Retrospectively analyzed the clinical data of the hospitalized 45 PT patients of an ear surgeon in one hospital between January 2013 to January 2020, and observed the effectiveness with surgery and non-surgery therapy. Surgical procedures include reconstruction the bone wall of sigmoid sinus by transmastoid approach and ligation of mastoid emissary vein. Non-surgery therapy includes anti-anemia therapy and observation. All patients have been followed-up in ENT outpatient. RESULTS: Of 45 cases, female : male was 43: 2, the mean age was 42.7 years old. The other PT patients were the subjective tinnitus except two females were the objective tinnitus. Of 40 cases, 38 patients underwent transmastoid approach to reconstructed sigmoid sinus bone wall, including 6 patients with the ligated mastoid emissary vein at the same period.The other 2 cases with the ligated mastoid emissary vein only.Five cases were treated by non-surgery therapies, including 2 cases anti-anemia therapy and 3 cases observation. The longest follow-up period was seven and a half years, the shortest was six months. One case was lost to follow up. The total cure rate was 80.0%(36/45), the surgery cure rate was 82.5%(33/40), the non-surgery cure rate was 60.0% (3/5). CONCLUSION: The pathophysiologic mechanism of the PT is still complex and unclear until now. However, the following conditions probably play an important role in the etiology: female, common features of anatomy anomalies, hemodynamic variations. It is a key point to confirm the responsible site or the main cause of the PT. Although the surgery is relatively simple, the effect is remarkable and no major postoperative complications, surgery could not be a only choice.

Background: Cancer poses a serious threat to human health and survival, and studies had been reported that imidazole or pyridine analogs play as an anti-cancer agent in cancer treatment. Meanwhile, Autophagy plays a dual and substantial role in maintaining cellular homeostasis in cancers, for it is either initiated to rescue cancer cells under stress or executed to promote autophagy cell death under certain circumstances. Objective: TIP-6 was designed and synthesized (7-(4-methoxyphenyl)-5,8α-diphenyl-1,2,3,7,8, 8α-hexahyd-roimidazo[1,2-α]pyridine-6) for evaluation of its biological effects on HepG2 cells and exploring the potential anti-cancer effect. Methods and Results: Chemical synthesis results indicated that the expected compound was obtained. The results of the MTT assay showed that TIP-6 arrested the growth of HepG2 cells in G2/M phase in the cell cycle, showing significant anti-proliferation effect. And analysis of morphological changes and formation of acidic vesicular organelles showed that the autophagy was induced but not apoptosis. The results were further validated by the enhanced expression of LC3I/II, Beclin1and down-regulated expression of Bcl-2in western blot analysis. In addition, the molecular docking predicted that TIP-6 preferentially binds to Bcl-2 and Bcl-xL in the active sites. Conclusion: Overall, this study demonstrated that autophagy cell death was executed in HepG2 cells which were induced by TIP-6.

lncRNA DLEU1 as a non-coding gene, involves in the occurrence and development of multiple tumors. However, there is no related report in endometrial carcinoma. In order to focus on the role and mechanism of lncRNA DLEU1 in endometrial carcinoma, we used qRT-PCR to detect the expression of lncRNA DLEU1 and found that lncRNA DLEU1 was highly expressed in endometrial carcinoma compared to normal endometrium. Moreover, compared to Ishikawa and KLE, lncRNA DLEU1 was higher in HEC-1B. In addition, up-regulation of lncRNA DLEU1 promoted cell viability, migration, invasion, and reduced the proportion of apoptosis. Otherwise, down-regulation of lncRNA DLEU1 produced opposite results. Xenograft nude mice model assay showed that lncRNA DLEU1 can promote tumorigenesis in vivo. RiP confirmed that lncRNA DLEU1 could bind to mTOR. The rescue experiments revealed that silence of mTOR after up-regulation of lncRNA DLEU1 resulted in decrease of cell viability, migration, and invasion and increase of apoptosis. The expression changes of PI3K, AKT1, p70S6K, rpS6, GSK3β, STAT3, and Bcl-xl were consistent with lncRNA DLEU1 and mTOR in Western blot. Thus, we suggest that lncRNA DLEU1 combines with mTOR and then increases the expression of PI3K/AKT/mTOR pathway to promote endometrial carcinoma tumorigenesis and progression. The present discovery has probability to provide a biomarker and lay the foundation for targeted therapy of endometrial carcinoma.

To evaluate the ability for the detection of 5 metal elements in serum in the laboratories of disease prevention and control system.The samples for calcium, magnesium, iron, copper and zinc detection were distributed to 48 laboratories of disease prevention and control system. Inductively coupled plasma mass spectrometry( ICP-MS) analysis or self-selected determination method were allowed to use during detection for each laboratory. The results were analyzed by robust statistical analysis and Z value was used to evaluate the detection ability.Of the laboratories involved in the study, 40 reported results of metal elements detection. Among them, 29 laboratories had satisfactory results, and 11 laboratories had unsatisfactory or suspicious results. The laboratory pass rate of this inter-laboratory comparison was60. 4%.The detection level of calcium, magnesium, iron, copper and zinc in serum in disease prevention and control system is generally satisfactory, but the detection ability of some laboratories needs to be further improved.

Fenoxaprop-p-ethyl (FPE) was studied for possible ecotoxicity on two representative toxigenic cyanobacteria including Microcystis aeruginosa and Microcystis viridis. Growth curves, chlorophyll a content, protein content, microcystin levels, oxidative stress, and apoptosis rates were measured for the two cyanobacteria after exposure to different concentrations of FPE. Results showed that the changes in chlorophyll a content and protein content were consistent with cell density, and M. viridis was more sensitive than M. aeruginosa to FPE. The results of oxidative stress indicated that FPE induced the generation of malondialdehyde (MDA) and enhanced the activities of antioxidant enzymes including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in these two cyanobacteria. To further explore the toxicity of FPE, apoptosis rates and toxin levels were measured for the two cyanobacteria. Different degrees of apoptosis rates were observed in the two cyanobacteria, and the apoptosis rates increased with the increase concentration of FPE. The intracellular and extracellular MC-LR were both affect by FPE. The presence of FPE in aquatic ecosystem may stimulate the synthesis and release of MC-LR, which may cause serious water pollution and pose threats to human health. These results may be useful for the ecotoxicity assessment of FPE and guiding the rational use of pesticides in agriculture.

Glyphosate has been used extensively for weed control in agriculture in many countries. However, glyphosate can be transported into the aquatic environment and might cause adverse effects on aquatic life. This study investigated the physiological characteristics of cyanobacteria Microcystis aeruginosa (M. aeruginosa) after exposure to glyphosate, and the results showed that changes in cell density production, chlorophyll a and protein content are consistent. In M. aeruginosa, oxidative stress caused by glyphosate indicated that 48h of exposure increased the concentration of malondialdehyde (MDA) and enhanced the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD). To further investigate the toxicity of glyphosate on M. aeruginosa, the viability of treated cells was monitored and the toxin release was determined. The results indicated that glyphosate induced apoptosis of and triggered toxin release in M. aeruginosa. These results are helpful for understanding the toxic effects of glyphosate on cyanobacteria, which is important for environmental assessment and protection. These results are also useful for guidance on the application of this type of herbicide in agricultural settings.

Purpose– The purpose of this paper is to investigates how organizational learning, absorptive capacity, cultural integration, specialization of the acquired firm and characteristics of transferred knowledge impact innovation performance subsequent to overseas acquisitions.Design/methodology/approach– Survey responses from 222 Chinese multinational enterprises engaged in overseas acquisitions.Findings– Differences between acquiring and acquired firms’ capabilities, while having a positive direct influence, suppress the positive impact of organizational learning and absorptive capacity, suggesting that multinationals require some basic level of capabilities to appropriate value from overseas acquisitions.Research limitations/implications– This paper investigates the impact of knowledge-seeking overseas acquisition of Chinese multinationals on innovation performance, as this appears to be the primary motive for making such acquisitions.Practical implications– Knowledge-seeking overseas acquisition should be based upon the absorptive capacity of the acquiring firm and complementarity between both firms. In knowledge-seeking overseas acquisitions, establishing an effective organizational learning mechanism is necessary for improving innovation performance.Originality/value– This paper reports on the behaviour and innovation performance of Chinese multinationals through analysis of primary data.

Natural compounds derived from plants have been an important source of numerous clinically useful anticancer agents. Nevertheless, limited studies indicate that xanthohumol (XN), a major prenylated flavonoid in hop plants (Humulus lupulus), may possess anticarcinogenic properties. The purpose of the present study was to clarify the antitumorigenic effects and the underlying mechanism of XN on breast cancer in vivo and in vitro. A 4T1 breast tumor mouse model was used in the present study to investigate XN suppression of tumor growth as detected by tumorigenicity assays in vivo. In addition, in vitro studies revealed that XN significantly decreased cell viability, induced G0/G1 cell cycle arrest and apoptosis in MCF-7 and MDA-MB-231 cells, as confirmed by an MTT assay, flow cytometry and western blot analysis, indicating anticarcinogenic activity of XN against breast cancer. Furthermore, immunohistochemistry was performed to confirm the inactivation of the Notch signaling pathway, Notch 1 and Ki-67, in vivo; consistently, XN caused decreased activation of the Notch signaling pathway and apoptotic regulators B-cell lymphoma-2 (Bcl-2), Bcl-extra large and caspase 3, as determined by western blot analysis in vitro. This study suggests that XN may potentially be useful as a chemopreventive agent during breast hyperplasia and carcinogenesis, acting via the regulation of Notch associated apoptotic regulators in vivo and in vitro.

Astrocytes perform several functions in the brain and spinal cord. Penicillin is commonly used for establishment of experimental epilepsy models. Previous studies have demonstrated that astragaloside IV (3-o-β-d-xylopyranosyl-6-o-β-d-glucopyranosyl-cycloastragenol; AS‑IV) has comprehensive pharmacological functions on the attenuation of inflammation. In the present study, primary astrocyte cell cultures were divided into three groups: Control group, penicillin (2,500 µM) treatment group (epilepsy model), and penicillin+AS‑IV (20, 40, 80 and 160 µmol/l) treatment group. The expression levels of inflammatory factors, including interleukin‑1β and tumor necrosis factor‑α, were determined in the groups using western blot and reverse transcription‑quantitative polymerase chain reaction analyses. The levels of members of the phosphorylated‑mitogen‑activated protein kinase (p‑MAPK) family, including p‑c‑Jun N‑terminal kinase 1/2, p‑extracellular signal‑regulated protein kinase 1/2 and p‑p38, were determined using western blot analysis. Cell viability of the astrocytes was detected using a 3‑(4,5‑dimethyl‑2‑thiazolyl)‑2,5‑diphenyl‑2‑H‑tetrazolium bromide assay and cell proliferation was evaluated using a Cell Counting Kit‑8 assay. The results revealed that AS‑IV significantly suppressed the expression of penicillin‑induced inflammatory factors in the astrocytes at the transcriptional and translational levels, and occurred in a dose‑dependent manner. The penicillin‑induced increase in the protein levels of the the p‑MAPK family were notably decreased by AS‑IV. In addition, the penicillin‑induced downregulation of primary astrocyte viability/cell proliferation was significantly reversed by the administration of AS‑IV. From these results, it was concluded that AS‑IV suppressed the penicillin‑induced upregulation of inflammatory factors and p‑MAPK in astrocytes, ultimately attenuating epilepsy.

The JAK2/STAT3 signaling pathway plays a critical role in oncogenesis and malignancy, which makes it a promising anticancer target. We report four N(6)-substituted adenosine analogues (AAs) as potential JAK2/STAT3 inhibitors identified through a STAT3-based high-throughput drug screening system. These AAs exhibited selective anti-cancer activity on human cancer cells and xenograft tumors with constitutively activated STAT3. They rapidly and potently suppressed constitutive and IL-6/IFN-γ-induced JAK2/STAT3 signal activation. In addition, we finally proved that the STAT3 signal blockage by three of these AAs was dependent on specific JAK2 inhibition. These AAs may represent new targeted therapeutic agents for JAK2/STAT3 hyper-activated human cancers.

The global usage of veterinary antibiotics is significant. These antibiotics can be released into the aquatic environment and exert toxic effects on non-target organisms. To explore the physiological effects of tetracycline antibiotics on aquatic life, the growth characteristics of and toxin release from the cyanobacterium Microcystis aeruginosa (M. aeruginosa) were studied. Results showed that the toxicity order of the three target antibiotics was TC (tetracycline) > CTC (chlortetracycline hydrochloride) > OTC (oxytetracycline) in terms of inhibition occurrence time and the EC10 and EC25 values. Further, the target antibiotics regulated the production of MC-LR (microcystin-LR) to different degrees. CTC destroyed the M. aeruginosa cells and resulted in a decreased MC-LR release but stimulated the ability to synthesise MC-LR. OTC had a relatively weaker toxicity compared with CTC, while TC was the most toxic among the three antibiotics. Therefore, TC is friendly to the aquatic environment because it simultaneously reduced the intracellular and extracellular MC-LR content. These results aid our understanding of the effects of tetracycline antibiotics on Microcystis aeruginosa, which is important for environmental evaluation and protection. These results are also helpful for guiding the application of veterinary antibiotics in agricultural settings.

As a component of NURD histone deacetylase complex, ZIP serves as a tumor suppressor gene in the development of breast tumors. However, whether it takes part in chemotherapy resistance remains poorly defined. In the present study, we reported that ZIP enhanced the response to SERM chemotherapy in ER-negative cells. Overexpression of ZIP suppressed EGFR expression level and restored ERalpha protein level in cells resistant to Tamoxifen. In vivo data confirmed those in vitro findings.

Ferrocene compounds are a class of biologically active compounds that has antitumour and antifungal properties. This study investigated the induction of apoptosis in human fibrosarcoma cells (HT1080) after treatment with a series of 6-ferrocenyl-3-subsituted7H-1,2,4-triazolo[3,4-b]- 1,3,4-thiadiazine (FTFs). We found that FTFs could suppress the viability of HT1080 cells. Cell cycle analysis showed that proliferative inhibition of HT1080 cells occurred through apoptosis, as the cells were blocked in G1 phase. Moreover, mitochondrial membrane staining assay demonstrated that FTFs exposure significantly decreased mitochondrial membrane potential. Finally, under the stress of FTFs, Bax/Bcl-2 ratio in HT1080 cells was significantly increased. These results suggested that FTFs-induced apoptosis in HT1080 cells may work dependent on a Bax/Bcl-2 pathway.

At harvest traits such as seed weight are the sum of development and responses to stresses over the growing season and particularly during the reproductive phase of growth. The aim here was to measure quantitative trait loci (QTL) underlying the seed weight from early development to drying post harvest. One hundred forty-three F(5) derived recombinant inbred lines (RILs) developed from the cross of soybean cultivars 'Charleston' and 'Dongnong 594' were used for the analysis of QTL underlying mean 100-seed weight at six different developmental stages. QTL x Environment interactions (QE) were analyzed by a mixed genetic mode based on 3 years' data. At an experiment-wise threshold of a=0.05 and by single-point analysis 94 QTL unaffected by QE underlay the mean seed weight at different developmental stages. Sixty-eight QTL affected by QE that also underlay mean seed weight were identified. From the 162 QTL 42 could be located on 12 linkage groups by composite interval mapping (LOD>2.0). The numbers, locations and types of the QTL and the genetic effects were different at each developmental stage. On linkage group C2 the distantly linked QTL swC2-1, swC2-2 and swC2-3 each affected mean seed weight throughout the different developmental stages. The DNA markers linked to the QTL possessed potential for use in marker-assisted selection for soybean seed size. The identification of QTL with genetic main effects and QE interaction effects suggested that such interactions might significantly alter seed weight during seed development.