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Atmospheric nitrogen deposition can alter soil microbial communities and further impact the structure and function of forest ecosystems. However, most studies are focused on positive or negative effects after nitrogen addition, and few studies pay attention to its interruption. In order to investigate whether interruption after different levels of short-term N additions still benefit soil health, we conducted a 2-year interruption after a 4-year short-term nitrogen addition (10 and 20 kg N·hm−2·yr−1) experiment; then, we compared soil microbial diversity and structure and analyzed soil physicochemical properties and their correlations before and after the interruption in Larix olgensis forest soil in northeast China. The results showed that soil ecological stabilization of Larix olgensis forest further improved after the interruption compared to pre-interruption. The TN, C:P, N:P, and C:N:P ratios increased significantly regardless of the previous nitrogen addition concentration, and soil nutrient cycling was further promoted. The relative abundance of the original beneficial microbial taxa Gemmatimonas, Sphingomonas, and Pseudolabrys increased; new beneficial bacteria Ellin6067, Massilia, Solirubrobacter, and Bradyrhizobium appeared, and the species of beneficial soil microorganisms were further improved. The results of this study elucidated the dynamics of the bacterial community before and after the interruption of short-term nitrogen addition and could provide data support and a reference basis for forest ecosystem restoration strategies and management under the background of global nitrogen deposition.

Abstract Aim This study is to investigate the current status of knowledge, attitude and self‐reported practice in hospice care among nurses in Hainan, China, and then to analyse its influencing factors and mediating effects. This provides a basis for formulating scientific and standardized hospice care training programmes for nurses. Methods This cross‐sectional study investigated knowledge, attitude and self‐reported practice in hospice care among 1819 nurses in Hainan, China. Convenience sampling was used to select participants from 45 hospitals and nursing homes in 14 cities and counties from October to December 2021. A scale of knowledge, attitude and self‐reported practice of healthcare providers in hospice care (Chinese version) was administered to collect data during the study period. Statistical analyses, including t‐tests, one‐way ANOVA, post‐hoc analysis and multiple linear regression, assessed the status of knowledge, attitude and self‐reported practice of hospice care in nurses and identified influencing factors. The PROCESS macro program model 4.0 was employed to explore the mediating effect of attitude on knowledge and self‐reported practice in hospice care. Results Nurses in Hainan displayed low knowledge (mean = 7.68, SD = 3.53), moderate attitudes (mean = 88.13, SD = 12.10) and self‐reported practice (mean = 51.81, SD = 9.82) in hospice care. Current employment and willingness to engage in hospice care were significant factors influencing knowledge, attitude and self‐reported practice in hospice care. Attitude partially mediated the relationship between knowledge and self‐reported practice. Patient or Public Contribution This study focuses on nurses' knowledge, attitude and self‐reported practice in hospice care and does not directly involve patients or the public. However, the findings enhance hospice care provided to patients and the broader community by improving nurses' knowledge and skills. This study informs evidence‐based training programmes and interventions, benefiting those in need of hospice care services.

Abstract Background Metastasis, the leading cause of cancer-related death in patients diagnosed with ovarian cancer (OC), is a complex process that involves multiple biological effects. With the continuous development of sequencing technology, single-cell sequence has emerged as a promising strategy to understand the pathogenesis of ovarian cancer. Methods Through integrating 10 × single-cell data from 12 samples, we developed a single-cell map of primary and metastatic OC. By copy-number variations analysis, pseudotime analysis, enrichment analysis, and cell–cell communication analysis, we explored the heterogeneity among OC cells. We performed differential expression analysis and high dimensional weighted gene co-expression network analysis to identify the hub genes of C4. The effects of RAB13 on OC cell lines were validated in vitro. Results We discovered a cell subcluster, referred to as C4, that is closely associated with metastasis and poor prognosis in OC. This subcluster correlated with an epithelial–mesenchymal transition (EMT) and angiogenesis signature and RAB13 was identified as the key marker of it. Downregulation of RAB13 resulted in a reduction of OC cells migration and invasion. Additionally, we predicted several potential drugs that might inhibit RAB13. Conclusions Our study has identified a cell subcluster that is closely linked to metastasis in OC, and we have also identified RAB13 as its hub gene that has great potential to become a new therapeutic target for OC.

Abstract Background Ovarian cancer (OC) is one of the commonest and deadliest diseases that threaten the health of women worldwide. It is essential to find out its pathogenic mechanisms and therapeutic targets for OC patients. Although NUF2 (Ndc80 kinetochore complex component) has been suggested to play an important role in the development of many cancers, but little is known about its function and the roles of proteins that regulate NUF2 in OC. This study aimed to investigate the effect of NUF2 on the tumorigenicity of OC and the activities of proteins that interact with NUF2. Methods Oncomine database and immunohistochemical (IHC) staining were used to evaluate the expression of NUF2 in OC tissues and normal tissues respectively. Normal ovarian epithelial cell lines (HOSEpiC) and OC cell lines (OVCAR3、HEY、SKOV3) were cultured. Western blot was applied to analyze the expression of NUF2 in these cell lines. Small interfering RNA (siRNA) was used to silence the expression of NUF2 in OC cell lines, SKOV3 and HEY. Gene Set Variation Analysis (GSVA), Gene Set Enrichment Analysis (GSEA), the CCK-8 method, colony formation assay and flow cytometry were conducted to analyze the biological functions of NUF2 in vitro. OC subcutaneous xenograft tumor models were used for in vivo tests. Immunoprecipitation and mass spectrometry (IP/MS) were performed to verify the molecular mechanisms of NUF2 in OC. IP, immunofluorescence, IHC staining, and Gene Expression Profiling Interactive Analysis platform (GEPIA) were used to analyze the relationship between HNRNPA2B1 and NUF2 in OC cells. SiRNA was used to silence the expression of HNRNPA2B1 in SKOV3 cells, reverse transcription quantitative polymerase chain reaction (RT-qPCR) assay and western blot were used to detect the effect of HNRNPA2B1 on NUF2. GEPIA, The Cancer Genome Atlas (TCGA) database, GSEA and western blot were used to detect the potential signaling pathways related to the roles of HNRNPA2B1 and NUF2 in OC cells. Results Our results showed high NUF2 expression in OC tissues and OC cell lines, which was associated with shorter overall survival and progression-free survival in patients. NUF2 depletion by siRNA suppressed the proliferation abilities and induced cell apoptosis of OC cells in vitro, and impeded OC growth in vivo. Mechanistically, NUF2 interacted with HNRNPA2B1 and activated the PI3K/AKT/mTOR signaling pathway in OC cells. Conclusion NUF2 could serve as a prognostic biomarker, and regulated the carcinogenesis and progression of OC. Moreover, NUF2 may interact with HNRNPA2B1 by activating the PI3K/AKT/mTOR signaling pathway to promote the development of OC cells. Our present study supported the key role of NUF2 in OC and suggested its potential as a novel therapeutic target.

Summary: Evidence concerning PM1 exposure, maternal blood pressure (BP), and hypertensive disorders of pregnancy (HDP) is sparse. We evaluated the associations using 105,063 participants from a nationwide cohort. PM1 concentrations were evaluated using generalized additive model. BP was measured according to the American Heart Association recommendations. Generalized linear mixed models were used to assess the PM1-BP/HDP associations. Each 10 μg/m3 higher first-trimester PM1 was significantly associated with 1.696 mmHg and 1.056 mmHg higher first-trimester SBP and DBP, and with 11.4% higher odds for HDP, respectively. The above associations were stronger among older participants (> 35 years) or those educated longer than 17 years or those with higher household annual income (> 400,000 CNY). To conclude, first-trimester PM1 were positively associated with BP/HDP, which may be modified by maternal age, education level, and household annual income. Further research is warranted to provide more information for both health management of HDP and environmental policies enactment.

Background Aberrant expression of circular RNAs (circRNAs) contributes to the initiation and progression of pulmonary hypertension (PH). Hypoxia‐inducible factor (HIF) is a well‐known modulator of hypoxia‐induced PH. The role and underlying mechanism of circRNAs in the regulation of HIF expression remains elusive. Methods and Results We profiled pulmonary artery transcriptomes using RNA sequencing and screened circRNAs associated with hypoxia treatment. The expression of a novel circRNA, circ_chr11_67292179–67294612 (circ‐myh8), was increased by hypoxia in a time‐dependent manner. We evaluated the effects of circ‐myh8 overexpression by adeno‐associated virus or inhibition by short hairpin RNA on proliferation and cell cycling in mice and pulmonary artery smooth muscle cells. Overexpression of circ‐myh8 promotes PH under normoxia, and disruption of circ‐myh8 by short hairpin RNA mitigates PH in chronic hypoxic mice. Biologically, circ‐myh8 induces the proliferation and cell‐cycle progression of pulmonary artery smooth muscle cells in vivo and in vitro. Mechanistically, RNA pull‐down and RNA immunoprecipitation assays were used to examine the interaction of circRNAs with the binding protein KAT7 (lysine acetyltransferase 7). The acetylation level of lysine 5 of histone H4 in the transcriptional initiation region of HIF1α was determined by chromatin immunoprecipitation assay followed by reverse transcription‐quantitative polymerase chain reaction. Circ‐myh8 acts as a modular scaffold to recruit histone acetyltransferase KAT7 to the promoters of HIF1α, which elicits acetylation of lysine 5 of histone H4 in their promoters. Conclusions Our findings not only reveal the pivotal roles of circ‐myh8 in governing histone modification in anti‐PH treatment but also advocate triggering the circ‐myh8/KAT7/HIF1α pathway to combat PH.

BackgroundOvarian cancer (OC) is a fatal gynecological tumor with high mortality and poor prognosis. Yet, its molecular mechanism is still not fully explored, and early prognostic markers are still missing. In this study, we assessed carcinogenicity and clinical significance of family with sequence similarity 83 member D (FAM83D) in ovarian cancer by integrating single-cell RNA sequencing (scRNA-seq) and a prognostic model.MethodsA 10x scRNA-seq analysis was performed on cells from normal ovary and high-grade serous ovarian cancer (HGSOC) tissue. The prognostic model was constructed by Lasso-Cox regression analysis. The biological function of FAM83D on cell growth, invasion, migration, and drug sensitivity was examined in vitro in OC cell lines. Luciferase reporter assay was performed for binding analysis between FAM83D and microRNA-138-5p (miR-138-5p).ResultsOur integrative analysis identified a subset of malignant epithelial cells (C1) with epithelial-mesenchymal transition (EMT) and potential hyperproliferation gene signature. A FAM83D+ malignant epithelial subcluster (FAM83D+ MEC) was associated with cell cycle regulation, apoptosis, DNA repair, and EMT activation. FAM83D resulted as a viable prognostic marker in a prognostic model that efficiently predict the overall survival of OC patients. FAM83D downregulation in SKOV3 and A2780 cells increased cisplatin sensitivity, reducing OC cell proliferation, migration, and invasion. MiR-138-5p was identified to regulate FAM83D’s carcinogenic effect in OC cells.ConclusionsOur findings highlight the importance of miR-138 -5p/FAM83D/EMT signaling and may provide new insights into therapeutic strategies for OC.

Atmospheric nitrogen deposition affects the health of forest ecosystems by altering soil microbial activity. However, the effects of nitrogen addition levels, morphology and ecosystem type on whether nitrogen addition is beneficial or detrimental to soil health is controver-sial, and most studies have focused on the negative effects on microbial structure. Based on this, this study conducted a four-year experiment of nitrogen (NaNO3) addition at two levels (10 and 20 kg N hm−2·yr−1) in the understory soil of Larix olgensis in northeastern China to study soil microbial properties, soil enzyme activities, and to analyze soil physi-cochemical properties and the correlation between them. The results showed that nitrogen addition reduced soil pH and increased soil NH4+-N and NO3−-N contents, thus promoting the activities of Urease (Ure), Acid phosphatase (ACP) and N-Acetamidoglucosidase (NAG) and inhibiting the activity of Leucine aminopeptidase (LAP) in soil, further improving the diversity and richness of soil microorganisms and increasing the dominant taxa of beneficial microorganisms. This may be due to soil acidification caused by the addition of nitrogen, which increases the effectiveness of nitrogen in the soil, improving soil properties, moving soil health in a beneficial direction, promoting beneficial microbial activity, and making the soil more suitable for the growth of the acid-loving tree species L. olgensis. In general, N addition favored the development of soil bacterial communities and the maintenance of soil nutrient status, and had a positive effect on the soil nutrient status of L. olgensis. The results of this study may provide an important scientific basis for adaptive management of forest ecosystems in the context of global nitrogen deposition.

BackgroundSpontaneous abortion is one of the prevalent adverse reproductive outcomes, which seriously threatens maternal health around the world.ObjectiveThe current study is aimed to evaluate the association between maternal age and risk for spontaneous abortion among pregnant women in China.MethodsThis was a case-control study based on the China Birth Cohort, we compared 338 cases ending in spontaneous abortion with 1,352 controls resulting in normal live births. The main exposure indicator and outcome indicator were maternal age and spontaneous abortion, respectively. We used both a generalized additive model and a two-piece-wise linear model to determine the association. We further performed stratified analyses to test the robustness of the association between maternal age and spontaneous abortion in different subgroups.ResultsWe observed a J-shaped relationship between maternal age and spontaneous abortion risk, after adjusting for multiple covariates. Further, we found that the optimal threshold age was 29.68 years old. The adjusted odds ratio (95% confidence interval) of spontaneous abortion per 1 year increase in maternal age were 0.97 (0.90–1.06) on the left side of the turning point and 1.25 (1.28–1.31) on the right side. Additionally, none of the covariates studied modified the association between maternal age and spontaneous abortion (P > 0.05).ConclusionsAdvanced maternal age (>30 years old) was significantly associated with increased prevalence of spontaneous abortion, supporting a J-shaped association between maternal age and spontaneous abortion.

ObjectiveCongenital heart disease (CHD) is complex in its etiology. Its genetic causes have been investigated, whereas the non-genetic factor related studies are still limited. We aimed to identify dominant parental predictors and develop a predictive model and nomogram for the risk of offspring CHD.MethodsThis was a retrospective study from November 2017 to December 2021 covering 44,578 participants, of which those from 4 hospitals in eastern China were assigned to the development cohort and those from 5 hospitals in central and western China were used as the external validation cohort. Univariable and multivariable analyses were used to select the dominant predictors of CHD among demographic characteristics, lifestyle behaviors, environmental pollution, maternal disease history, and the current pregnancy information. Multivariable logistic regression analysis was used to construct the model and nomogram using the selected predictors. The predictive model and the nomogram were both validated internally and externally. A web-based nomogram was developed to predict patient-specific probability for CHD.ResultsDominant risk factors for offspring CHD included increased maternal age [odds ratio (OR): 1.14, 95% CI: 1.10–1.19], increased paternal age (1.05, 95% CI: 1.02–1.09), maternal secondhand smoke exposure (2.89, 95% CI: 2.22–3.76), paternal drinking (1.41, 95% CI: 1.08–1.84), maternal pre-pregnancy diabetes (3.39, 95% CI: 1.95–5.87), maternal fever (3.35, 95% CI: 2.49–4.50), assisted reproductive technology (2.89, 95% CI: 2.13–3.94), and environmental pollution (1.61, 95% CI: 1.18–2.20). A higher household annual income (100,000–400,000 CNY: 0.47, 95% CI: 0.34–0.63; > 400,000 CNY: 0.23, 95% CI: 0.15–0.36), higher maternal education level (13–16 years: 0.68, 95% CI: 0.50–0.93; ≥ 17 years: 0.87, 95% CI: 0.55–1.37), maternal folic acid (0.21, 95% CI: 0.16–0.27), and multivitamin supplementation (0.33, 95% CI: 0.26–0.42) were protective factors. The nomogram showed good discrimination in both internal [area under the receiver-operating-characteristic curve (AUC): 0.843] and external validations (development cohort AUC: 0.849, external validation cohort AUC: 0.837). The calibration curves showed good agreement between the nomogram-predicted probability and actual presence of CHD.ConclusionWe revealed dominant parental predictors and presented a web-based nomogram for the risk of offspring CHD, which could be utilized as an effective tool for quantifying the individual risk of CHD and promptly identifying high-risk population.

Abstract Background Epithelial ovarian cancer (EOC), as a lethal malignancy in women, is often diagnosed as advanced stages. In contrast, intermediating between benign and malignant tumors, ovarian low malignant potential (LMP) tumors show a good prognosis. However, the differential diagnosis of the two diseases is not ideal, resulting in delays or unnecessary therapies. Therefore, unveiling the molecular differences between LMP and EOC may contribute to differential diagnosis and novel therapeutic and preventive policies development for EOC. Methods In this study, three microarray data (GSE9899, GSE57477 and GSE27651) were used to explore the differentially expressed genes (DEGs) between LMP and EOC samples. Then, 5 genes were screened by protein–protein interaction (PPI) network, receiver operating characteristic (ROC), survival and Pearson correlation analysis. Meanwhile, chemical-core gene network construction was performed to identify the potential drugs or risk factors for EOC based on 5 core genes. Finally, we also identified the potential function of the 5 genes for EOC through pathway analysis. Results Two hundred thirty-four DEGs were successfully screened, including 81 up-regulated genes and 153 down-regulated genes. Then, 5 core genes (CCNB1, KIF20A, ASPM, AURKA, and KIF23) were identified through PPI network analysis, ROC analysis, survival and Pearson correlation analysis, which show better diagnostic efficiency and higher prognostic value for EOC. Furthermore, NetworkAnalyst was used to identify top 15 chemicals that link with the 5 core genes. Among them, 11 chemicals were potential drugs and 4 chemicals were risk factors for EOC. Finally, we found that all 5 core genes mainly regulate EOC development via the cell cycle pathway by the bioinformatic analysis. Conclusion Based on an integrated bioinformatic analysis, we identified potential biomarkers, risk factors and drugs for EOC, which may help to provide new ideas for EOC diagnosis, condition appraisal, prevention and treatment in future.

BackgroundThe outbreak of coronavirus disease 2019 (COVID-19) has endangered human health and life. This pandemic has changed people's lifestyle and affected the regular delivery of standard cancer treatment. In the present study, we aimed to explore the influencing factors of delayed treatment in patients with breast cancer during COVID-19 pandemic.MethodsThis study was a cross-sectional investigation, and the subjects were patients who were discharged from the department of burn and plastic surgery after February 2020. All participants completed this study's online questionnaire based on the WeChat and Wenjuanxing platforms. Levels of anxiety and depression were measured by the Hospital Anxiety and Depression Scale (HADS). Patients were divided into a delay group and non-delay group according to the occurrence of delayed treatment. Univariate analysis was performed by using the t test or chi-square test. A logistic regression model was employed to determine factors associated with delayed treatment.ResultsThe present study included a total of 397 patients with breast cancer, among whom delayed treatment occurred in 76 patients, accounting for 19.1%. Scores on both the anxiety subscale and depression subscale in delay group were significantly higher than those in non-delay group. Compared with non-delay group, we found that patients in delay group usually had a higher level of education (P = 0.020), worse self-feeling (P = 0.030), poor compliance of medical order (P = 0.042), and a higher prevalence of anxiety (P = 0.004) and depression (P = 0.012). Traffic inconvenience was also an important relevant factor for delayed treatment (P = 0.001). The prevalence of recurrence in delay group was higher than that in non-delay group (P = 0.018). By using logistic multivariate regression analysis, the results revealed that level of education and traffic inconvenience were independent factors influencing delayed treatment in patients with breast cancer during COVID-19 pandemic.ConclusionThe prevalence of delayed treatment in patients with breast cancer during COVID-19 pandemic is relatively high. Our findings reveal several influencing factors closely associated with delayed treatment, which is useful information that will be beneficial for patients to receive standardized therapy by taking targeted measures.

Cyclin Y (CCNY) is a novel cyclin and highly conserved in metazoan species. Previous studies from our and other laboratory indicate that CCNY play a crucial role in tumor progression. There are two CCNY isoform which has different subcellular distributions, with cytoplasmic isoform (CCNYc) and membrane distribution isoform (CCNYm). However, the expression and function of CCNY isoforms is still unclear. We firstly found CCNYc was expressed in natural lung cancer tissue and cells through the subcellular distribution. Co-IP and immunofluorescence showed that both CCNYm and CCNYc could interact with PFTK1. Further studies illustrated that CCNYc but not CCNYm enhanced cell migration and invasion activity both in vivo and vitro. The function of CCNYc could be inhibited by suppression of PFTK1 expression. In addition, our data indicated that tropomyosin 4 (TPM4), a kind of actin-binding proteins, was down-regulated by suppression of CCNY. F-actin assembly could be controlled by CCNYc as well as PFTK1 and TPM4. As a result, CCNY was mainly expressed in lung cancer. CCNYc could promote cell motility and invasion. It indicated that CCNYc/PFTK1 complex could promote cell metastasis by regulating the formation of F-actin via TPM4.

We propose a novel fiber-optic auditory nerve of ground (FANG) in the suburb based on the fiber-optic distributed vibration sensor (DVS). The feasibility and effectiveness of the principle prototype FANG for traffic flow monitoring are proved and investigated by the field experiment. One of the 31.8 km-long redundant optical fiber of the buried optical-fiber cable for data transmission is utilized as the sensing fiber. Then, the phase-sensitive optical time-domain reflectometer (φ-OTDR) based DVS is realized and regarded as the FANG. The vibration events at 9 observation points with different ground conditions along the sensing fiber are detected by a threshold algorithm during 6.5 hours from 8:00 am. Then, the vibration events are analyzed in combination with the ground conditions to recognize the machine working in the factory, rammer working and the vehicles passed through near different areas and roads. The traffic flow is estimated by the vibration-counting with a counting error that is believed to be in an acceptable range. The distribution and the fluctuation trends of the estimated traffic flow are useful and enlightening for the traffic monitoring and pre-warning of special events, such as an accident. The accuracy can be improved by artificial intelligence methods in the future. It seems that our proposed FANG can be a potential and effective tool for the internet of things, smart ground and smart traffic in the suburb where the video and other information collection methods are not available.

Angle is an important concept in geometry. Young children have difficulty separating angle size from other dimensions such as the length of angle sides, perhaps due to whole-object bias in word learning. The present study used the pre-test–training–post-test design to investigate the effectiveness of two ways of separating angle from angle size in 3–6-year-old Chinese preschoolers. A total of 228 children were given a pre-test and 219 of them failed the crucial test. 168 of the 219 children were present at school during the training phase and were randomly assigned to three groups: the “toma” group (n = 57), which received training to call the whole angle figure as “toma” and angle size as angle size; the “angle/angle size” group (n = 56), which received the training of separating “angle” from “angle size”; and the control group (n = 55), which used “angle size” alone to represent both the overall angle figure and angle size. Results showed that the “toma” group improved significantly more than the other two groups, the latter of which did not differ from each other. These results suggest that it is insufficient to have two separate words/phrases (angle and angle size) for children to learn to differentiate angle from angle size, perhaps due to their shared usage of the word angle. Instead, the use of a novel term is necessary and sufficient to improve learning. Implications for preschool education are discussed.

Optical trapping is widely used in different areas, ranging from biomedical applications, to physics and material sciences. In recent years, optical fiber tweezers have attracted significant attention in the field of optical trapping due to their flexible manipulation, compact structure, and easy fabrication. As a versatile tool for optical trapping and manipulation, optical fiber tweezers can be used to trap, manipulate, arrange, and assemble tiny objects. Here, we review the optical fiber tweezers-based trapping and manipulation, including dual fiber tweezers for trapping and manipulation, single fiber tweezers for trapping and single cell analysis, optical fiber tweezers for cell assembly, structured optical fiber for enhanced trapping and manipulation, subwavelength optical fiber wire for evanescent fields-based trapping and delivery, and photothermal trapping, assembly, and manipulation.

This paper presents a microfluidics-based approach capable of continuously characterizing instantaneous Young’s modulus (Einstantaneous) and specific membrane capacitance (Cspecific membrane) of suspended single cells. In this method, cells were aspirated through a constriction channel while the cellular entry process into the constriction channel was recorded using a high speed camera and the impedance profiles at two frequencies (1 kHz and 100 kHz) were simultaneously measured by a lock-in amplifier. Numerical simulations were conducted to model cellular entry process into the constriction channel, focusing on two key parameters: instantaneous aspiration length (Linstantaneous) and transitional aspiration length (Ltransitional), which was further translated to Einstantaneous. An equivalent distribution circuit model for a cell travelling in the constriction channel was used to determine Cspecific membrane. A non-small-cell lung cancer cell line 95C (n = 354) was used to evaluate this technique, producing Einstantaneous of 2.96 ± 0.40 kPa and Cspecific membrane of 1.59 ± 0.28 μF/cm2. As a platform for continuous and simultaneous characterization of cellular Einstantaneous and Cspecific membrane, this approach can facilitate a more comprehensive understanding of cellular biophysical properties.

Disregulation of dickkopf-related protein 1 (DKK1) has been reported in a variety of human cancers. However, how DKK1 functions in Non-small cell lung cancer has not been revealed. In the current study, DKK1 was knocked out by the lentivirus-mediated short hairpin RNA interference approach in H1299 and 95C non-small cell lung cancer cell lines. Subsequently, the migration and invasion ability were assessed by wound-healing and transwell assays. In addition, epithelial-mesenchymal transition markers and β-catenin were examined by Western blot analysis. The signaling pathway downstream of DKK1 was characterized using the Wnt signaling pathway inhibitor, IWP2, and glycogen synthase kinase 3 beta inhibitor, LiCl. Immunofluorescence analysis investigated the subcellular localization of β-catenin. The results suggested that knockdown of DKK1 caused reduced migration and invasion ability of H1299 and 95C cells. DKK1 silencing resulted in the downregulation of epithelial-mesenchymal transition-related proteins, such as Snail and zinc finger E-box binding homeobox 1. Besides, DKK1 silencing inhibited β-catenin and promoted the phosphorylation of β-catenin. Mechanism results indicated that the expression of β-catenin was reduced in H1299 or 95C cells after being treated with Wnt signaling inhibitor, IWP2. In addition, the inhibition of β-catenin phosphorylation by glycogen synthase kinase 3 beta inhibitor, LiCl, significantly enhanced the migration and invasion capacities in DKK1-knockdown cell lines. Furthermore, cell immunofluorescence revealed that nuclear β-catenin was reduced when DKK1 was knocked down. Taken together, these findings suggest that DKK1 induces the occurrence of epithelial-mesenchymal transition and promotes migration and invasion in non-small cell lung cancer cells. Mechanically, β-catenin plays a vital role in DKK1-induced non-small cell lung cancer cell migration and invasion, and DKK1 inhibits the phosphorylation of β-catenin, resulting in the increased nuclear localization of β-catenin.

A novel cyclin, cyclin Y, is one of the most highly conserved members of the cyclin superfamily, which is famous for their important roles in regulating the cell cycle and transcription. Recently, it was found that cyclin Y was up-regulated in many cancers. Cyclin Y may play a crucial role in tumor progression. The function and molecular mechanism of cyclin Y protein will be discussed in this paper.

Triptolide is a diterpenoid compound that inhibits the inflammation of rheumatoid arthritis (RA). However, the use of triptolide is limited due to its strong gastrointestinal toxicity. The purpose of this work was to develop a transdermal delivery system for triptolide to reduce this toxicity. A microemulsion-based hydrogel (MBH) was prepared from the combination of Gemseal 40-oleic acid as oil phase, Tween 80-labrasol as surfactant, anhydrous ethanol as co-surfactant, water as aqueous phase and Poloxamer 407 as hydrogel matrix. Rheological measurements, environmental scanning electron microscopy (ESEM) and transdermal experiments in vitro were used to characterize triptolide-loaded and blank MBH preparations. The effects of Poloxamer 407 and triptolide on the rheological properties and microstructures of the MBH were determined. Transparent and homogeneous MBH could only be formed when the concentration of Poloxamer 407 in the selected O/W microemulsion was in the range of 14.0–16.0% (w/w). When the concentration of Poloxamer 407 increased, the rheological properties such as the yield stresses (σy), storage and loss moduli (G′, G″) of the formulations increased, and the network structures became more compact. The addition of triptolide did not change the interconnected network structures of the MBH preparations. MBH preparations afford a better sustained release profile when compared to microemulsions, a finding confirmed by an in vitro permeation test in mice. MBH appears to be a promising vehicle for transdermal delivery of triptolide.

As increases in hepatocyte growth factor/scatter factor (HGF/SF) induce retinal pigment epithelial (RPE) migration and proliferation into the vitreous cavity and contribute to proliferative vitreoretinopathy (PVR) development, we determined if changes in miR-182 expression affect such behavioral changes. We found that miR-182 expression was less in PVR clinical samples than in primary RPE cells whereas c-Met was upregulated. Ectopic miR-182 inhibited RPE cell proliferation, cell cycle, and migration. Bioinformatic analysis identified c-Met as a miR-182 target, which was confirmed with the luciferase reporter assay. Transfection of miR-182 into RPE cells induced c-Met downregulation, which led to reduced cell proliferation and migration through declines in p-Akt formation. MiR-182 downregulation along with c-Met upregulation in PVR tissues suggest that these two opposing effects play important roles in PVR development. As ectopic miR-182 expression suppressed RPE cell proliferation and migration, strategies to selectively upregulate miR-182 expression in a clinical setting may provide a novel option to treat this disease.

Background and objective It has been proved that mTOR was an important signal transduction molecular and protein kinase regulating cell growth and proliferation, and mTOR could activate the downstream protein effector. PTEN could negatively regulate mTOR signal pathway and inhibit its activity. The aim of this study is to detect the mRNA expression levels of mTOR and PTEN gene, which are the key genes of mTOR signaling pathway in human non-small cell lung cancer (NSCLC) tissue. The relationship between mTOR signaling pathway and NSCLC is also explored. Methods Lung cancer tissue specimens were obtained from 65 patients. Adjacent-tumor non-small cell lung cancer tissues from the 30 patients were served as control. The RT-PCR technique was used to detect the mTOR and PTEN gene expression levels. Results The average mRNA expression levels of mTOR gene were significantly higher (0.23±0.16) in lung cancer than in adjacent-tumor tissue (0.12±0.09)(P < 0.01). The average mRNA expression levels of PTEN gene were (0.19±0.28) in lung cancer, while the mRNA expression levels of PTEN gene were (0.53±0.28) in adjacent-tumor tissue (P < 0.01). The levels of PTEN gene expression in non-small cell lung cancer were significantly lower than that in adjacent-tumor lung tissue. There are not significant relationship between mTOR and PTEN gene expression levels and patients’ age, gender, pathological type, differentiation, lymph node metastasis, except tumor size. Conclusion The expression of mTOR is activated in NSCLC. The expression of PTEN is absent or decreased. The mTOR activated in NSCLC may be correlate with the absent or decreased of PTEN. The absent or decreased expression of PTEN and the actived mTOR may play important roles in carcinogenesis and metastasis of NSCLC.

The transcription factor PAX6 is primarily expressed in embryos. PAX6 is also expressed in several tumors and plays an oncogenic role. However, little is known about the role of PAX6 in lung cancer.The function of PAX6 in lung cancer cells was evaluated by small interfering RNA-mediated depletion of the protein followed by analyses of cell proliferation, anchorage-independent growth, and cell cycle arrest. The changes of cyclin D1, pRB, ERK1/2, p38 expression caused by PAX6 inhibition were detected using western-blotting. The PAX6 mRNA level in 52 pairs of tumors and corresponding matched adjacent normal tissues from non-small cell lung cancer patients and lung cancer cell lines was detected by real-time PCR.Suppression of PAX6 expression inhibited cell growth and colony formation in A549 and H1299 cells. The percentage of cells in G1-phase increased when PAX6 expression was inhibited. The cyclin D1 protein level, as well as the pRB phosphorylation level, decreased as a result of PAX6 down-regulation. The activity of ERK1/2 and p38 was also suppressed in PAX6 knock-down cells. The PAX6 mRNA was highly expressed in lung cancer tissue and lung cancer cell lines. In most patients (about 65%), the relative ratio of PAX6 mRNA in primary NSCLC versus adjacent tissues exceeded 100.Our data implicated that PAX6 accelerates cell cycle progression by activating MAPK signal pathway. PAX6 mRNA levels were significantly elevated in primary lung cancer tissues compared to their matched adjacent tissues.