WRKY transcription factors play a key role in the tolerance of biotic and abiotic stresses across various crop species, but the function of some WRKY genes, particularly in tomato, remains ...unexplored. Here, we characterize the roles of a previously unstudied WRKY gene, SlWRKY8, in the resistance to pathogen infection and the tolerance to drought and salt stresses. Expression of SlWRKY8 was up‐regulated upon Pseudomonas syringae pv. tomato DC3000 (Pst. DC3000), abiotic stresses such as drought, salt and cold, as well as ABA and SA treatments. The SlWRKY8 protein was localized to the nucleus with no transcription activation in yeast, but it could activate W‐box‐dependent transcription in plants. The overexpression of SlWRKY8 in tomato conferred a greater resistance to the pathogen Pst. DC3000 and resulted in the increased transcription levels of two pathogen‐related genes SlPR1a1 and SlPR7. Moreover, transgenic plants displayed the alleviated wilting or chlorosis phenotype under drought and salt stresses, with higher levels of stress‐induced osmotic substances like proline and higher transcript levels of the stress‐responsive genes SlAREB, SlDREB2A and SlRD29. Stomatal aperature was smaller under drought stress in transgenic plants, maintaining higher water content in leaves compared with wild‐type plants. The oxidative pressure, indicated by the concentration of hydrogen peroxide (H2O2) and malondialdehyde (MDA), was also reduced in transgenic plants, where we also observed higher levels of antioxidant enzyme activities under stress. Overall, our results suggest that SlWRKY8 functions as a positive regulator in plant immunity against pathogen infection as well as in plant responses to drought and salt stresses.
The effects of moderate salinity on the responses of woody plants to UV-B radiation were investigated using two Populus species (Populus alba and Populus russkii). Under UV-B radiation, moderate ...salinity reduced the oxidation pressure in both species, as indicated by lower levels of cellular H2O2 and membrane peroxidation, and weakened the inhibition of photochemical efficiency expressed by O-J-I-P changes. UV-B-induced DNA lesions in chloroplast and nucleus were alleviated by salinity, which could be explained by the higher expression levels of DNA repair system genes under UV-B&salt condition, such as the PHR, DDB2, and MutSα genes. The salt-induced increase in organic osmolytes proline and glycine betaine, afforded more efficient protection against UV-B radiation. Therefore moderate salinity induced cross-tolerance to UV-B stress in poplar plants. It is thus suggested that woody plants growing in moderate salted condition would be less affected by enhanced UV-B radiation than plants growing in the absence of salt. Our results also showed that UV-B signal genes in poplar plants PaCOP1, PaSTO and PaSTH2 were quickly responding to UV-B radiation, but not to salt. The transcripts of PaHY5 and its downstream pathway genes (PaCHS1, PaCHS4, PaFLS1 and PaFLS2) were differently up-regulated by these treatments, but the flavonoid compounds were not involved in the cross-tolerance since their concentration increased to the same extent in both UV-B and combined stresses.
We carried out a genome-wide association study among Chinese women to identify risk variants for breast cancer. After analyzing 607,728 SNPs in 1,505 cases and 1,522 controls, we selected 29 SNPs for ...a fast-track replication in an independent set of 1,554 cases and 1,576 controls. We further investigated four replicated loci in a third set of samples comprising 3,472 cases and 900 controls. SNP rs2046210 at 6q25.1, located upstream of the gene encoding estrogen receptor α (ESR1), showed strong and consistent association with breast cancer across all three stages. Adjusted odds ratio (95% CI) were 1.36 (1.24-1.49) and 1.59 (1.40-1.82), respectively, for genotypes A/G and A/A versus G/G (P for trend 2.0 × 10−15) in the pooled analysis of samples from all three stages. We also found a similar, albeit weaker, association in an independent study comprising 1,591 cases and 1,466 controls of European ancestry (Ptrend = 0.01). These results strongly implicate 6q25.1 as a susceptibility locus for breast cancer.
The grain growth retardation mechanism and the effect of cooling rate on VC-doped WC–Co cemented carbides were investigated in this work.WC–30Co and WC–30Co–VC were prepared by powder ...metallurgy,liquid-phase sintering at 1400 ℃ and followed by water quenching（150 ℃/s） or furnace cooling（＊0.083 ℃/s）.Based on the results of electron probe microanalysis（EPMA）,we found that WC concentration in the Co binder was independent of VC doping during liquid-phase sintering,hence barely contributing to the retardation of WC grain growth.In contrast,the（W,V）Cx phase formed at the WC/Co interfaces played a major role in retarding WC grain growth during liquid-phase sintering.The effect of cooling rate on the morphology of（W,V）Cxwas revealed by high-resolution transmission electron microscopy（HRTEM） and energy-dispersive spectroscopy（EDS）.In the water-quenched WC–30Co–VC,（W,V）Cxprecipitates were found as thin layers at the WC/Co interfaces.In contrast,both thin layers of similar thickness and nanoparticles of（W,V）Cx were observed in the furnace-cooled counterpart.These observations listed above suggested that thin（W,V）Cxlayers were stable structures effectively suppressing the growth of WC grains and their thickness remained independent of the cooling rate.The（W,V）Cxnanoparticles,however,may be inhibited through rapid cooling,ensuring the VC-doped WC–Co cemented carbides desired toughness.
Background
The efficacy of adjuvant targeted therapy for operable lung cancer is still under debate. Comprehensive genetic profiling is needed for detecting co‐mutations in resected epidermal growth ...factor receptor (EGFR)‐mutated lung adenocarcinoma (ADC), which may interfere the efficacy of adjuvant tyrosine kinase inhibitor (TKI) treatment.
Materials and Methods
Mutation profiling of 416 cancer‐relevant genes was conducted for 139 resected stage I–IIIa lung ADCs with EGFR mutations using targeted next‐generation sequencing. Co‐mutation profiles were systematically analyzed.
Results
Rare EGFR alterations other than exon 19 deletion and L858R, such as L861Q (∼3%) and G719A (∼2%), were identified at low frequencies. Approximately 10% of patients had mutations in EGFR exon 20 that could confer resistance to first‐generation TKIs. Ninety‐one percent of patients harbored at least one co‐mutation in addition to the major EGFR mutation. TP53 was the top mutated gene and was found more frequently mutated at later stage. Markedly, NF1 mutations were found only in stage II–III ADCs. Conversely, RB1 mutations were more frequent in stage I ADCs, whereas APC mutations were observed exclusively in this group. Thirty‐four percent of patients with EGFR TKI‐sensitizing mutations had genetic alterations involving EGFR downstream effectors or bypass pathways that could affect the response to EGFR TKIs, such as PIK3CA, BRCA1, and NOTCH1.
Conclusion
Operable lung ADCs with EGFR TKI‐sensitizing mutations are associated with a high proportion of co‐mutations. Mutation profiling of these resected tumors could facilitate in determining the applicability and efficacy of adjuvant EGFR TKI therapeutic strategy.
Implications for Practice
The efficacy of adjuvant epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) therapy for lung cancer harboring EGFR mutation after surgical resection is still under debate. Next‐generation sequencing of 416 cancer‐relevant genes in 139 resected lung cancers revealed the co‐mutational landscape with background EGFR mutation. Notably, the study identified potential EGFR TKI‐resistant mutations in 34.71% of patients with a drug‐sensitizing EGFR mutation and who were naive in terms of targeted therapy. A comprehensive mutation profiling of these resected tumors could facilitate in determining the applicability and efficacy of adjuvant EGFR TKI therapeutic strategy for these patients.
摘要
背景 辅助靶向治疗对可手术肺癌的疗效仍在争论中。在可切除的表皮生长因子受体 (EGFR) 突变肺腺癌 (ADC) 中检测共突变需要全面的遗传分析，这可能会影响辅助酪氨酸激酶抑制剂 (TKI) 治疗的疗效。
材料和方法 使用靶向的下一代测序技术，对 139 个具有 EGFR 突变的可切除的 I‐IIIa 期肺 ADC 进行416 个癌症相关基因突变分析。系统地分析了共突变谱。
结果 除了外显子 19 缺失和 L858R 外的罕见EGFR 改变，如 L861Q(~3%) 和 G719A(~2%)，较低频率被发现。大约 10% 的患者发生 EGFR 外显子 20 突变，这可能导致对第一代 TKI 耐药。除主要EGFR突变外，91% 的患者出现至少一种共突变。TP53是最常见的突变基因，在后期的突变频率较高。显然的是，NF1突变只在 II‐III 期 ADC 中发现。相反，RB1 突变在 I 期 ADC 中更为频发，而仅在该组中观察到 APC 突变。34% EGFR TKI 敏感突变患者的遗传变异涉及 EGFR 下游效应靶点或旁路途径，可能影响对 EGFR TKI 的反应，如PIK3CA、BRCA1，和NOTCH1。
结论 具有 EGFR TKI 敏感突变的可手术肺 ADC 与高比例的共突变相关。这些可切除肿瘤的突变谱有助于确定辅助 EGFR TKI 治疗策略的适用性和有效性。
实践意义:辅助表皮生长因子受体 (EGFR) 酪氨酸激酶抑制剂 (TKI) 治疗肺癌术后EGFR突变的疗效仍存在争议。在 139 例可切除的肺癌中，对 416 个癌症相关基因进行的下一代测序揭示了EGFR突变背景下的共突变图景。值得注意的是，该研究在 34.71% 的药物敏感性EGFR突变患者中发现了潜在的 EGFR TKI 耐药突变，而这些患者缺乏靶向治疗方面的经验。对这些可切除肿瘤进行全面的突变谱分析有助于确定辅助 EGFR TKI 治疗策略对这些患者的适用性和有效性。
The best treatment option for patients with operable lung cancer is debatable. In this study, a comprehensive mutation profiling was performed on resected EGFR‐mutated lung adenocarcinoma using next‐generation sequencing that targeted cancer‐relevant genes to identify potential candidates for adjuvant tyrosine kinase inihibitors treatment post‐operation.
Creatine kinase plays a key role in the energy homeostasis of vertebrate cells. Creatine kinase B (CKB), a cytosolic isoform of creatine kinase, shows upregulated expression in a variety of cancers. ...In this research, we confirmed that some ovarian cancer tissues had elevated CKB expression at the protein level. The functions of CKB in ovarian cancer progression were investigated in the ovarian cancer cell line Skov3, which has a high CKB expression. It was found that CKB knockdown inhibited Skov3 cell proliferation and induced apoptosis under hypoxia or hypoglycemia conditions. CKB depletion also sensitized Skov3 to chemotherapeutic agents. Furthermore, the CKB knockdown reduced glucose consumption and lactate production, and increased ROS production and oxygen consumption. This suggested that CKB knockdown decreased cytosolic glycolysis and resulted in a tumor suppressive metabolic state in Skov3 cells. Consequently, we found that the knockdown of CKB induced G2 arrest in cell cycle by elevating p21 expression and affected the PI3K/Akt and AMPK pathways. These findings provide new insights in the role of CKB in cancer cell survival and tumor progression. Our results also suggest that CKB depletion/inhibition in combination with chemotherapeutic agents might have synergistic effects in ovarian cancer therapy.
Abstract Background Cerebral metastases are the main determining factor in the failure of locally advanced non-small-cell lung cancer (NSCLC) management. Our study assessed the risk factors of brain ...metastases in patients with postoperative, locally advanced NSCLC. Implications for PCI treatment are discussed. Methods Two hundred twenty-three patients treated with surgical resection for stage III-N2 NSCLC were retrospective analyzed to elucidate risk factors for development of brain metastases, and to establish a mathematical model. Results Median survival time for this patient population was 29.5 months. Frequency of brain metastases in the entire patient population was 38.1% (85/223). Frequency of brain metastases in patients with single mediastinal lymph-node region with metastases at 1, 2, and 3 years was 5.6%, 14.0%, and 19.0%, respectively. The frequency of brain metastases in patients with multiple mediastinal lymph-node regions with metastases was 31.8%, 60.3%, 68.0%, respectively ( P < 0.001). The frequency of brain metastases among patients with mediastinal metastasis number less than 4, 4–6, and more than 6 was significantly different ( P < 0.001). There were also significant differences in brain metastases frequency between patients with complete versus incomplete resection ( P = 0.001), and patients with non-squmous versus squamous ( P = 0.029), and patients administered adjuvant chemotherapy versus none ( P = 0.032). Conclusion A mathematical model to predict brain metastases risk was developed. It can aid in selection of patients with locally advanced NSCLC for PCI in clinical trails.
The relationship between the 6-minute walk test (6MWT) and pulmonary function test in stable chronic obstructive pulmonary disease (COPD) remains unclear. We evaluate the correlation of 6MWT and ...spirometric parameters in stable COPD with different severities. 6MWT data assessed included three variables: the 6-minute walk distance (6MWD), 6-minute walk work (6MWORK), and pulse oxygen desaturation rate (SPO(2)%).
6MWT and pulmonary function test were assessed for 150 stable COPD patients with different severities. Means and standard deviations were calculated for the variables of interest. Analysis of variance was performed to compare means. Correlation coefficients were calculated for 6MWT data with the spirometric parameters and dyspnea Borg scale. Multiple stepwise regression analysis was used to screen pulmonary function-related predictors of 6MWT data.
The three variables of 6MWT all varied as the severities of the disease. The 6MWD and 6MWORK both correlated with some spirometric parameters (positive or negative correlation; the absolute value of r ranging from 0.34 to 0.67; P < 0.05) in severe and very severe patients, and the SPO2% correlated with the dyspnea Borg scale in four severities (r = -0.33, -0.34, -0.39, -0.53 respectively; P < 0.05). The 6MWD was correlated with the 6MWORK in four severities (r = 0.56, 0.57, 0.72, 0.81 respectively, P < 0.05), and neither of them correlated with the SPO(2)%. The percent of predicted forced expiratory volume in 1 second (FEV(1)% predicted) and residual volume to total lung capacity ratio (RV/TLC) were predictors of the 6MWD, and the maximum voluntary ventilation (MVV) was the predictor of the 6MWORK.
6MWT correlated with the spirometric parameters in severe and very severe COPD patients. 6MWT may be used to monitor changes of pulmonary function in these patients.
Protein disulfide isomerase (PDI) functions as an isomerase to catalyze thiol:disulfide exchange, as a chaperone to assist protein folding, and as a subunit of prolyl-4-hydroxylase and microsomal ...triglyceride transfer protein. At a lower concentration of 0.2 μm, PDI facilitated the aggregation of unfolded rabbit muscle creatine kinase (CK) and exhibited anti-chaperone activity, which was shown to be mainly due to the hydrophobic interactions between PDI and CK and was independent of the cross-linking of disulfide bonds. At concentrations above 1 μm, PDI acted as a protector against aggregation but an inhibitor of reactivation during CK refolding. The inhibition effect of PDI on CK reactivation was further characterized as due to the formation of PDI-CK complexes through intermolecular disulfide bonds, a process involving Cys-36 and Cys-295 of PDI. Two disulfide-linked complexes containing both PDI and CK were obtained, and the large, soluble aggregates around 400 kDa were composed of 1 molecule of tetrameric PDI and 2 molecules of inactive intermediate dimeric CK, whereas the smaller one, around 200 kDa, was formed by 1 dimeric PDI and 1 dimeric CK. To our knowledge this is the first study revealing that PDI could switch its conformation from dimer to tetramer in its functions as a foldase. According to the observations in this research and our previous study of the folding pathways of CK, a working model was proposed for the molecular mechanism of CK refolding catalyzed by PDI.
To evaluate the potential reconsideration of curative operative treatment for patients with unresectable stage IIIA (N2) non-small-cell lung cancer (NSCLC).
From Jan. 1999 to Dec. 2002, 76 patients ...with unresectable stage IIIA (N2) NSCLC were entered in this study. They had all been proved by chest CT, chest film and fiberobronchoscopy. Twenty-one (27.6%) patients were examined by mediastinoscopy. All the patients received two cycles of chemotherapy with NVB (25 mg/m(2), D1, D5) and carboplatin (300 mg/m(2), D1). All the patients were staged again three weeks after induction chemotherapy. Sixty-four patients who achieved partial response (PR) or complete response (CR) were allowed to undergo surgery. Twelve patients who did not responde to chemotherapy received radiotherapy instead. Of the 64 surgically treated patients, 56 (84.7%) had a complete resection and then received 2 cycles of chemotherapy using the same regime, 8 patients had an incomplete resection and then received radiotherapy for the residual tu
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