OTSUCNV: an adaptive segmentation and OTSU-based anomaly classification method for CNV detection using NGS data.

Copy-number variations (CNVs), which refer to deletions and duplications of chromosomal segments, represent a significant source of variation among individuals, contributing to human evolution and being implicated in various diseases ranging from mental illness and developmental disorders to cancer. Despite the development of several methods for detecting copy number variations based on next-generation sequencing (NGS) data, achieving robust detection performance for CNVs with arbitrary coverage and amplitude remains challenging due to the inherent complexity of sequencing samples. In this paper, we propose an alternative method called OTSUCNV for CNV detection on whole genome sequencing (WGS) data. This method utilizes a newly designed adaptive sequence segmentation algorithm and an OTSU-based CNV prediction algorithm, which does not rely on any distribution assumptions or involve complex outlier factor calculations. As a result, the effective detection of CNVs is achieved with lower computational complexity. The experimental results indicate that the proposed method demonstrates outstanding performance, and hence it may be used as an effective tool for CNV detection.

Effects of biochar application on the loss characteristics of Cd from acidic soil under simulated rainfall conditions.

Biochar is widely used for immobilizing heavy metals in soil as a kind of high-effective passivator. This research conducted incubation and simulated rainfall experiments to study the effects of biochar application on the loss characteristics of runoff and sediment, as well as the transportation of the Cd during the water erosion process. Two rainfall intensities (60 and 120 mm h-1) and five biochar application rates (0%, 1%, 3%, 5%, and 7%) were considered in the experiment. The result showed that slaking had a greater effect than mechanical stirring in aggregate breakdown of the soil, and the addition of biochar generally increased the sensitivity of the soil to wet stirring, while had no obvious influence on the resistance to slaking. The H2O and CaCl2 extractable Cd in soil significantly decreased with the increase of biochar application rate. The runoff yields decreased with the increase of biochar application rate at both the two rainfall intensities, while the eroded sediment generally decreased at the 120 mm h-1 rainfall intensity. The addition of biochar tended to increase the loss of the middle-sized (1-0.05 mm) aggregates at the 60 mm h-1 rainfall intensity, whereas reduced their loss at the 120 mm h-1 rainfall intensity. Biochar application could significantly reduce the concentration of Cd in the runoff and decreased the total loss amount of Cd (sediment+runoff) in most of the cases. Excessively high level (7%) of biochar application may aggravate soil erosion and result in more Cd loss.

[Temporal and Spatial Variation Characteristics and Source Analysis of Agricultural Non-point Source Pollution Load in Guangdong During the Past 20 Years].

The agricultural non-point source pollution (ANPSP) load in Guangdong province is very large and has a serious impact on the regional ecological environment. Inventory analysis was used to assess and analyze the temporal and spatial variation characteristics of the ANPSP load of Guangdong province during 1999-2019, and the sources of ANPSP were discussed, as well as the relationship between them. The results showed that, during the past 20 years, the total ANPSP pollution loads of Guangdong province decreased by 6.08%, and the pollution loads of chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP) increased by -11.88%, 4.99%, and 26.17%, respectively. The input intensity of chemical fertilizers and pesticides increased by 112.19% and 60.38%, respectively. The Pearl River Delta had the highest ANPSP loads in Guangdong province, followed by those in northern, western, and eastern Guangdong. Livestock and poultry breeding were the main sources of COD, the total percent fertilizers and livestock and poultry breeding were the main sources of TN, and livestock and poultry breeding and aquaculture were the main sources of TP. In addition, the contribution of pollutants discharged from aquaculture showed an obvious increasing trend. There were certain differences in the pollution sources in different regions. In western Guangdong, northern Guangdong, and eastern Guangdong, livestock and poultry breeding were the main sources of COD and TP, and fertilizer was the main source of TN; by contrast in the Pearl River Delta, aquaculture had become the main source of TN and TP pollution loads. The correlation results showed that the decline in the total ANPSP in Guangdong province was mainly due to the increase in high urbanization rate and the decrease in the proportion of rural population. In general, there were stage changes in the time and differences in spatial characteristics and sources of ANPSP in Guangdong province. A combination of comprehensive treatment and targeted pollution treatment should be adopted, and fertilizer reduction measures and pollution treatment in the aquaculture should be strengthened in an all-around way, focusing on strengthening the treatment of pollution from aquaculture in the Pearl River Delta region and the treatment of rural life pollution in northern Guangdong.

Radiomics signature on CECT as a predictive factor for invasiveness of lung adenocarcinoma manifesting as subcentimeter ground glass nodules.

Controversy and challenges remain regarding the cognition of lung adenocarcinomas presented as subcentimeter ground glass nodules (GGNs). Postoperative lymphatic involvement or intrapulmonary metastasis is found in approximately 15% to 20% of these cases. This study aimed to develop and validate a radiomics signature to identify the invasiveness of lung adenocarcinoma appearing as subcentimeter ground glass nodules. We retrospectively enrolled 318 subcentimeter GGNs with histopathology-confirmed adenocarcinomas in situ (AIS), minimally invasive adenocarcinomas (MIA) and invasive adenocarcinomas (IAC). The radiomics features were extracted from manual segmentation based on contrast-enhanced CT (CECT) and non-contrast enhanced CT (NCECT) images after imaging preprocessing. The Lasso algorithm was applied to construct radiomics signatures. The predictive performance of radiomics models was evaluated by receiver operating characteristic (ROC) analysis. A radiographic-radiomics combined nomogram was developed to evaluate its clinical utility. The radiomics signature on CECT (AUC: 0.896 [95% CI 0.815-0.977]) performed better than the radiomics signature on NCECT data (AUC: 0.851[95% CI 0.712-0.989]) in the validation set. An individualized prediction nomogram was developed using radiomics model on CECT and radiographic model including type, shape and vascular change. The C index of the nomogram was 0.915 in the training set and 0.881 in the validation set, demonstrating good discrimination. Decision curve analysis (DCA) revealed that the proposed model was clinically useful. The radiomics signature built on CECT could provide additional benefit to promote the preoperative prediction of invasiveness in patients with subcentimeter lung adenocarcinomas.

Prevalence trends in non-alcoholic fatty liver disease at the global, regional and national levels, 1990-2017: a population-based observational study.

OBJECTIVE: Non-alcoholic fatty liver disease (NAFLD) is the leading cause of end-stage liver diseases worldwide. Understanding NAFLD prevalence and trends over time at the global, regional and national levels is critical to understanding the NAFLD disease burden and creating more tailored prevention strategies. DESIGN: Population-based observational study. SETTING: The study was global, including 21 regions and 195 countries or territories. MAIN OUTCOMES MEASURE: The estimated annual percentage change for NAFLD prevalence. RESULTS: Worldwide, cases of NAFLD have increased from 391.2 million in 1990 to 882.1 million in 2017, with the prevalence rate increasing from 8.2% to 10.9% during the same period. The increasing trends were consistent across sexes. Case numbers were highest in East Asia, followed by South Asia, then North Africa and the Middle East. The highest prevalence of NAFLD was observed in North Africa and the Middle East, while the greatest increase was detected in Western Europe, followed by Tropical Latin America, then high-income North America. CONCLUSION: Nearly all countries or territories worldwide have experienced a significant increase in NAFLD prevalence. The greatest increase was observed in Oman. Almost all countries showed a significant increasing trend in NAFLD prevalence over the past three decades. This drastic increase is alarming and suggests that NAFLD has emerged as a new public health concern worldwide. As such, more efficient prevention strategies are urgently needed.

Mortality trends of liver cancer among patients with type 2 diabetes at the global and national level.

BACKGROUND: Primary liver cancer (PLC) is a commonly diagnosed malignancy, especially in developing countries. Diabetes is one of the well-determined risk factors for PLC. We aimed to describe the temporal trends of PLC mortality among diabetic patients. METHODS: We retrieved the PLC mortality data among diabetic patients from the Global Burden of Disease (GBD) study 2017 online database. Estimated average percentage change (EAPC) was used to quantify the PLC age-standardized mortality rate (ASMR) trends, by sex and country, between 1990 and 2017. RESULTS: Globally, the number of PLC related deaths increased from 3732.1 in 1990 to 9506.4 in 2017, with the ASMR increased from 0.09/100,000 to 0.12/100,000 (EAPC = 0.98, 95% CI 0.82, 1.14) among diabetic patients. Both the ASMR of PLC and its temporal trend were highly heterogeneous across the world. Between 1990 and 2017, a total of 135, 19, and 41 countries or territories experienced a significant increase, remained stable, and experienced a significant decrease in PLC ASMR, respectively. The greatest increase was mainly detected in developed countries, such as the USA, the UK, and Australia. By contrast, the most pronounced decrease was majorly found in developing regions. CONCLUSIONS: In diabetic patients, the PLC mortality was significantly increased at the global level and in approximately 70% of countries or territories over the last three decades. The increasing trend indicated that diabetes is an increasingly important risk factor for PLC and suggested that more tailored prevention strategies are needed for each country.

Cetuximab enhances the efficiency of irinotecan through simultaneously inhibiting the MAPK signaling and ABCG2 in colorectal cancer cells.

BACKGROUND: The present study sought to investigate the combined effects of cetuximab and irinotecan on colorectal cancer cells as well as the mechanisms underlying their anti-cancer effects. MATERIAL AND METHODS: High performance liquid chromatography, Hoechst staining assay, and western blotting analysis were used to detect intracellular drug concentrations, cell apoptosis, and protein expression in the presence of cetuximab, irinotecan, and the combination of both. RESULTS: Cetuximab was found to increase intracellular concentrations of irinotecan as well as cytotoxicity by inhibiting the epidermal growth factor receptor and, by extension, the downstream RAS-RAF-MEK-ERK signaling pathway. Cetuximab therefore induced apoptosis and improved the effect of irinotecan in colorectal cancer cells. It was also shown that cetuximab inhibited the drug efflux activity of ABCG2. In combination with irinotecan, cetuximab can both significantly induce cell apoptosis by inhibiting the RAS-RAF-MEK-ERK signaling pathway and improve the effects of irinotecan by decreasing drug efflux through the inhibition of ABCG2. CONCLUSION: These features contribute to its anti-cancer potential.

Use of contrast-enhanced computed tomographic imaging to diagnose and evaluate Behçet's disease with vascular complications.

In the present study, multi-slice CT results of patients with Behçet's disease (BD) and vascular complications were retrospectively evaluated. From January 2016 to May 2018, 45 of 361 patients with BD were diagnosed with vascular involvement. The clinical background, laboratory parameters and response to therapy of those patients were assessed. The following characteristics of vascular aneurysms were analyzed: Maximum diameter, length, wall thickness, borders, luminal changes, mural thrombus, cystic change of the vessel walls, asymmetric bulging of the right part of the aortic wall (RP type) and calcific plaques. The 45 BD patients analyzed included 37 males and 8 females with a median age of 40 years (30-49 years). Significant differences were observed among genders regarding age, ocular disorders and digestive-tract ulceration. A total of 42 aneurysms were identified with a mean diameter of 43 mm. Most aneurysmal walls (88%) were homogeneously enhanced on contrast-enhanced CT. Comparison of groups classified by aortic and larger arterial aneurysms indicated that aneurysms occurring in the aorta were more likely to form a mural thrombus, have a thicker wall (P<0.001) and unclear borders (P=0.036), to be of the RP type (P=0.003) and have a longer extension (P=0.001) compared with those in larger arteries. Unclear border of the aneurysmal wall was the only radiologic predictor correlated with an elevated erythrocyte sedimentation rate (P<0.001). In conclusion, characteristic CT imaging features of aneurysms may help to diagnose vascular involvement of BD and assess its severity, particularly in the absence of the classical clinical manifestations.

Regulatory effect of Act1 on the BAFF pathway in B-cell malignancy.

The aim of the present study was to ascertain whether nuclear factor (NF)-κB Activator 1 (Act1) was involved in B cell-activating factor (BAFF) regulation in B-cell malignancy. The human B-cell malignancy cell lines Raji, Daudi and BALL-1 were cultured and the expression of BAFF receptor (BAFF-R) mRNA and protein was analyzed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting, respectively. NF-κB signaling was also assessed using western blotting. Act1 silencing was performed using Act1 small interfering RNA. BAFF-R levels were assessed using flow cytometry. It was demonstrated that BAFF-R was upregulated in all three cell lines and RT-qPCR, and western blotting confirmed these results. Act1 overexpression was demonstrated to induce BAFF-R upregulation, whereas Act1 knockdown resulted in BAFF-R downregulation. Furthermore, the NF-κB pathway was activated by Act1 overexpression and inhibited following Act1 knockdown. The results of the present study demonstrated that Act1 can regulate BAFF via targeting NF-κB signaling, which suggests that Act1 may be a promising therapeutic target for the treatment of B-cell malignancy.

H19 contributes to poor clinical features in NSCLC patients and leads to enhanced invasion in A549 cells through regulating miRNA-203-mediated epithelial-mesenchymal transition.

Recent studies have demonstrated that the overexpression of H19 may contribute towards development of tumorigenesis in various types of cancer. To investigate the role of H19 in the development of non-small cell lung cancer (NSCLC), 76 NSCLC tissues samples and their adjacent normal tissue samples were collected. Expression level of H19, and its association with clinicopathological features and overall survival was analyzed. It was found that compared with normal adjacent tissues, H19 expression was elevated in NSCLC tissues along with a decreased miR-203 expression level. It was also found that patients who were in advanced clinical stages had a higher H19 and a lower miR-203 expression compared to normal tissues. The overall survival time of patients with higher H19 expression was shorter compared with the lower H19 expression group. Upregulation of A549 enhanced cell proliferation and promoted invasion. Overexpression of H19 stimulated the epithelial-mesenchymal transition (EMT) process in lung cancer cells and demonstrated typical morphological characteristics of EMT. The level of mesenchymal marker protein, such as Vimentin and SNAI1 increased; while CDH1 protein level decreased. Also, H19 negatively regulated miR-203. Inhibition of H19 attenuated miR-203 induced EMT process. Upregulation of H19 contributes to poor clinical features in patients with NSCLC, induces occurrence of EMT, promotes proliferation and stimulates cell invasion in NSCLC cell line through regulating miRNA-203 mediated EMT.

Low-dose CT examination for rib fracture evaluation: A pilot study.

The aim of this study was to assess the applicability of low-dose thoracic computed tomography (CT) in the diagnosis of rib fractures.A total of 37 trauma patients were selected for CT scanning using a noise index (NI) model. Each patient was scanned at both NI = 11 and NI = 26, while the other scanning parameters were kept the same. The scanning dose length product (DLP) and effective dose (ED) were recorded after each examination. Two radiologists diagnosed the rib fractures by degree (I, II, III, and IV) using Bone Reading software and axial images. Image quality was scored by 2 experienced radiologists using a 5-point scale. The numbers and degrees of rib fractures for different NIs were recorded and tested using the Chi-squared test. The interobserver differences were determined by kappa statistics.The CTDIvols and EDs for NI = 11 and NI = 26 were 9.82 ±â€Š4.78, 5.75 ±â€Š2.75, and 2.14 ±â€Š1.19 and 1.24 ±â€Š0.73, respectively; the latter was decreased by 78.2% and 78.4% relative to the former. Low-dose thoracic CT was feasible for the auxiliary diagnosis of rib fractures using Bone Reading software (P > .05). There was perfect interobserver concordance in terms of diagnostic acceptability (kappa = 0.931, 0.905).The use of an appropriate low-dose CT scanning technique is satisfactory for the assessment and diagnosis of rib fractures.

Serum microRNA-15a level acts as a potential diagnostic and prognostic biomarker for human esophageal squamous cell carcinoma.

BACKGROUND: Clinical significance of microRNA (miR)-15a in human esophageal squamous cell carcinoma (ESCC) remains unclear. OBJECTIVE: To evaluate the expression level of miR-15a and to determine its potential for diagnosis and prognosis in ESCC. METHODS: Quantitative reverse transcription polymerase chain reaction was performed to examine the expression levels of miR-15a in ESCC tissues and patients' sera. The diagnostic and prognostic implications of serum miR-15a level in human ESCC were further evaluated. RESULTS: Expression levels of miR-15a in ESCC tissues and patients' sera were significantly decreased (both P< 0.001). Additionally, serum miR-15a had an optimal diagnostic cut-off point (2.29) for ESCC with sensitivity of 86.36% and specificity of 100.00%. Moreover, low serum miR-15a level more frequently occurred in ESCC patients with advanced tumor-node-metastasis, T and N stages (all P= 0.01) and poor tumor differentiation (P= 0.03). The Kaplan-Meier curve showed that low miR-15a expression was significantly associated with shorter overall survival (OS) and disease-free survival (DFS) of ESCC patients (both P< 0.001). Further multivariate analysis identified miR-15a as an independent prognostic factor for both OS and DFS (both P= 0.01). CONCLUSION: Decreased expression of miR-15a may play a crucial role in ESCC development and progression. Serum miR-15a level could be used as a potential diagnostic and prognostic marker in clinics.

[Mechanism of Regulating BAFF Signaling Pathway by Act1 in B Cell Lymphoma].

OBJECTIVE: To explore the mechanism of regulating B cell-activating factor (BAFF) signalling pathway by NF-κB activator 1 (Act1) in B cell lymphoma so as to provide a new thinking for treatment of B cell lymphoma. METHODS: The human B cell lymphoma cell lines including Raji, Daudi and BALL-1 were cultured, when the cells were in logarithmic phase, the RNA was extracted, and the Act1 was amplified by RT-PCR; and pTT5-Act1 expression plasmid was constructed and was transfected into cells; the Act1 was silenced by using Act1 mRNA; the NF-κB signaling pathway protein was detected by Western blot. RESULTS: After silence or overexpression of Act1, the proliferation levels of Raji, Daudi and BALL-1 cells were up- or down-regulated, respectively. Overexpression and silence of Act1 could down-or up-regulate BAFF-R expression level, furthermore could inhibit or activate of NF-κB signalling pahway, respectively. CONCLUSION: Among 3 above-mentioned B cell lymphoma cell lines, Act1 plays negative regulating role, indicating that the Act1 may be a promising therapeutic target for treating B cell lymphoma.

The lncRNA MALAT1 is a novel biomarker for gastric cancer metastasis.

The metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) is frequently over-expressed and serves as a prognostic marker in human cancers. However, little is known about the role of MALAT1 in gastric cancer. Here, we reported that the tissue and plasma MALAT1 levels were significantly higher in gastric cancer patients with distant metastasis (P<0.01) than patients without distant metastasis and the healthy controls. In addition, high levels of plasma MALAT1 independently correlated to a poor prognosis for gastric cancer patients (hazard ratio, 0.242; 95% CI, 0.154-0.836; P=0.036; Cox regression analysis). Functional studies revealed that knockdown of MALAT1 could inhibit cell proliferation, cell cycle progression, migration and invasion, and promote apoptosis in gastric cancer cells. Furthermore, the miR-122-IGF-1R signaling correlated with the dysregulated MALAT1 expression in gastric cancer. These data suggest that MALAT1 could function as an oncogene in gastric cancer, and high MALAT1 level could serve as a potential biomarker for the distant metastasis of gastric cancer.

Isoliquiritigenin prevents the progression of psoriasis-like symptoms by inhibiting NF-κB and proinflammatory cytokines.

UNLABELLED: Isoliquiritigenin (ISL) is an important flavonoid component of licorice and has been reported to possess anti-inflammatory and antioxidant properties, but its exact mechanism of action remains poorly understood. Previously, we demonstrated that ISL could suppress IL-6 expression in multiple myeloma. Here, we further characterized the anti-inflammatory effects of ISL in several psoriasis models, including the keratin 14/vascular endothelial growth factor (VEGF) transgenic mouse, the imiquimod (IMQ)-induced psoriasis-like mouse, and the human keratinocytes HaCaT and NHEK in vitro. We found that ISL ameliorated the inflammatory process in psoriasis models but not in their respective controls. Moreover, the anti-inflammatory effects of ISL were attributed to the suppression of nuclear factor-κB (NF-κB) activity, which consequently resulted in the reduction of pro-inflammation cytokines IL-6 and IL-8 expression. In conclusion, ISL exhibited anti-inflammatory effects in psoriasis models, by downregulating IL-6 and IL-8 via suppression of NF-κB activity, suggesting that ISL might serve as a potential candidate for treatment of psoriasis and other autoimmune inflammatory diseases. KEY MESSAGE: ISL could ameliorate the inflammatory process of psoriasis. ISL could suppress NF-κB and subsequent production of a series of pro-inflammatory cytokines. Dual-inhibitory activity against IL-6 and IL-8 of ISL is implemented via inhibiting NF-κB. ISL exerts no inhibitory effects on normal human keratinocytes or wild-type Balb/c mice, implying its low toxicity and safety.

CT features of lung scar cancer.

OBJECTIVE: To explore the CT features of lung scar cancer (LSC). METHODS: CT images of 41 LSCs and 66 non-LSCs were retrospectively compared in terms of location, size, shape, border, speculation, lobulation, pleural indentation, surrounding ground-glass opacification (sGGO), vessel convergence, vacuolation, calcification and satellite opacification. RESULTS: Thirty-eight LSCs were histopathologically identified as adenocarcinoma. The LSCs and non-LSCs were located 8.73±8.65 and 12.55±10.67 mm from the pleura, respectively. The mean lesion sizes (3-D ratios) in the initial LSC, pre-surgical LSC and non-LSC images were 24.28±6.29 (0.33±0.65), 32.23±8.14 (0.60±0.18) and 23.24±3.73 (0.35±0.61) mm, respectively. The initial and pre-surgical LSC images showed significant differences in speculation and sGGO (P<0.05). Significant differences were also noted in vacuolation, vessel convergence and sGGO between the pre-surgical LSC and the non-LSC images (P<0.05) and in vacuolation between the initial LSC and the non-LSC images (P<0.05). CONCLUSIONS: Despite similar CT features of LSCs and non-LSCs, the early detection and diagnosis of LSCs is possible by studying scar-tissue changes such as enlargement and sGGO associated with well-defined lesion borders in follow-up CT images.

MicroRNA expression profiles associated with acquired gefitinib-resistance in human lung adenocarcinoma cells.

The aim of the present study was to establish, characterize and elucidate the potential mechanisms of acquired gefitinb resistance, using the A549 human lung cancer cell line. A gefitinib-resistant A549 sub-clone was established by exposure to escalating gefitinib concentrations over a period of 16-24 months. Half maximal inhibitory concentration (IC50) values were quantified using a real time cytotoxicity assay. The expression profiles of the parent and resistant sub-clone A549 cells were detected using the µParaflo® Microfluidics Biochip microRNA (miRNA) Microarray. The ArrayPro software was used to analyze the differential expression levels of the miRNA, and bioinformatics software was used to predict the potential target genes of the differentially expressed miRNAs. Quantitative polymerase chain reaction (qPCR) was used to confirm the results of the miRNA microarray. A miRNA mimic was transfected into the gefitinib-resistant cells, in order to predict target gene interaction effects, following gefitinib treatment. Protein expression level differences were confirmed by western-blot analysis. Real time cytotoxicity assays revealed a 3-fold increase in the IC50 values of the gefitinib-resistant sub-clones, as compared with the parent cells. There were marked morphological differences between the parent and resistant cells. In the microarray analysis, the gefitinib-resistant sub-clones had 25 upregulated and 18 downregulated miRNAs, as compared with the parent cells. The qPCR revealed that miR-7 was significantly downregulated, which was concordant with the results of the microarray. The results of the present study suggest that miR-7 may significantly improve the sensitivity of cancer cells to gefitinib. The data presented in the present study provides an experimental basis and theory that miRNAs may be involved in acquired gefitinib-resistance of lung adenocarcinoma, and miR-7 may have potential clinical effects in the reversal of drug resistance.

Induced IGF-1R activation contributes to gefitinib resistance following combined treatment with paclitaxel, cisplatin and gefitinib in A549 lung cancer cells.

Gefitinib demonstrates excellent performance in the treatment of lung adenocarcinoma patients; yet, there was no added benefit in combination with chemotherapy as reported in a phase III clinical trial. For exploring the mechanism of the failed combination therapy in lung cancer, in the present study, four therapy assessment groups, including a control group, a chemotherapy group [paclitaxel+cisplatin (TP)], a gefitinib monotherapy group (G) and a combination group[paclitaxel+cisplatin+gefitinib (TP+G)], were established in an A549 cell line and mouse xenotransplanted tumor models.By HPLC, we found that the gefitinib concentration was significantly higher in the combination group when compared to that in the G group in the non-small cell lung cancer cell line, A549 (p<0.05). Following the treatment time extension,an increased cell growth rate was observed in the combination group, while the cellular concentration of gefitinib was not decreased. The expression levels of P-IGF-1R, P-SRC and P-ERK in the fourth combination treatment group were significantly higher than levels in the fourth G treatment and control groups (p<0.05). Following downregulating of IGF-1R in the fourth combination treatment group, drug sensitivity was recovered in vitro. In the mouse model, compared with the gefitinib monotherapy group, the combination group exhibited a smaller tumor volume, lower body weight and reduced survival rate (p<0.05). Gefitinib concentrations in the serum and tumor tissues in the combination therapy group were also decreased when compared with these concentrations in the gefitinib alone group. The present study is the first to demonstrate that the decreased gefitinib concentration in serum and tumor tissues is one of the reasons resulting in the failed combination treatment (chemotherapy+gefitinib) in vivo study. Frequent use of the combination treatment in A549 lung cancer cells induced IGF-1R activation which contributed to gefitinib resistance and gave rise to the failure of the combination therapy.

CD44 acts through RhoA to regulate YAP signaling.

The Hippo pathway plays an important role in both physical and pathogenesis processes. As crucial downstream effectors of Hippo pathway, YAP is inhibited by Lats1/2 through phosphorylation. However, upstream signals that regulate the Hippo pathway have been still poorly understood. Here, we found that knockdown of CD44 reduced YAP expression and nuclear localization, but nearly had no effect on its upstream effectors, Mst1 and Lats1. Downregulated CD44 expression also significantly decreased the expression of YAP downstream effectors CTGF, Cyr61 and EDN1 at mRNA level. Our next study showed that knockdown of CD44 inhibited RhoA expression, which was consistent with RhoA knockdown mediated YAP downregulation. Furthermore, we demonstrated that over expression of the constitutively active RhoA (RhoA-V14) could block the YAP expression decrease mediated by CD44 knockdown. Moreover, downregulation of CD44 significantly promoted cell apoptosis and inhibited cell proliferation, cell cycle progression and migration, which were consistent with the effects of RNAi-mediated YAP knockdown in both A549 and HepG2 cells. Overall, data are presented showing that CD44 could act through RhoA signaling to regulate YAP expression and this study also provide new insights into the regulatory mechanisms of the Hippo-YAP pathway.

[CT diagnosis of different pathological types of ground-glass nodules].

OBJECTIVE: To explore the CT features of ground-glass nodules (GGN) including preinvasive lesions [atypical adenomatous hyperplasia (AAH) and adenocarcinoma in situ (AIS)], minimally invasive adenocarcinoma (MIA) and invasive adenocarcinoma (IAC). METHODS: Ninety-seven GGN lesions confirmed by operation pathology were included in this study. The lesions were divided into three groups: preinvasive lesion group (24 cases), MIA group (39 cases), IAC group (34 cases). The lesion size, 3-dimensional ratio, 2-dimensional ratio in axial images, lesion density, shape, speculation, lobulation, air-containing space and pleural indentation on the preoperative CT images in the three groups were analyzed and compared with pathological results. The data were statistically analyzed using SPSS 17.0. RESULTS: All preinvasive lesions presented as pure GGN on CT image, most showed round-like shape, clear and smooth border. MIA presented as pure GGN or mixed GGN on CT image, most showed round-like shape, with a clear and smooth border. IAC most presented as mixed GGN on CT image, often showed irregular shape. Speculation, lobulation, air-containing space and pleural indentation displayed gradually increasing from preinvasive lesions to MIA and IAC. There were statistically significant differences in lesion size, CT density, shape, air-containing space, speculation, pleural indentation and long diameter of solid component between the MIA and IAC groups (P < 0.05 for all). There were statistically significant differences in CT density values and long diameters of solid component of the lesions between the preinvasive lesion group and MIA group (P < 0.05). The AUC of solid component of the preinvasive lesion group and MIA group was 0.705, and that of the MIA and IAC groups was 0.814. CONCLUSION: Comprehensive analysis of the CT image features of GGNs, especially the solid component in the lesions, may help to the preoperative and differential diagnosis of preinvasive lesions, MIA and IAC.