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Background/ObjectiveThe third-generation epidermal growth factor receptor (EGFR) -tyrosine kinase inhibitor (TKIs), such as osimertinib, designed for targeting the acquired drug-resistant mutation of EGFR T790M, was approved as the first-line therapy for advanced EGFR-mutated non-small cell lung cancer (NSCLC). Thus, detection of the EGFR T790M mutation for NSCLC is crucial. However, tissue samples are often difficult to obtain, especially in patients at advanced stages. This study assessed the performances of droplet digital polymerase chain reaction (ddPCR) and next-generation sequencing (NGS) in detecting EGFR T790M status and abundance in the plasma ctDNA samples of patients with NSCLC. We also explored the association between T790M status and abundance and the response to third-generation EGFR-TKIs.MethodsA total of 201 plasma samples with matched tissues, 821 plasma samples, and 56 patients who received third-generation EGFR-TKIs with response evaluation were included in this study. ddPCR and NGS were used to detect the mutation status and abundance of T790M in the tissues and/or blood samples.ResultsThe results showed that the sensitivity and the specificity of EGFR T790M mutation status detected by ddPCR in plasma samples were 81.82% and 91.85%, respectively, compared with the tissue samples, with a consistency coefficient of 0.740. Among the 821 plasma samples, the positive rates of EGFR T790M detected by ddPCR and NGS were 34.2% (281/821) and 22.5% (185/821), respectively. With NGS results as the reference, the sensitivity and the specificity of ddPCR were 100% and 84.91%, respectively, and the consistency coefficient of the two methods was 0.717. In addition, we found that a higher EGFR T790M abundance was linked to a higher treatment response rate to the third-generation EGFR-TKIs regardless of the classification of the median value of 0.43% (P = 0.016) or average value of 3.16% (P = 0.010).ConclusionTaking these data together, this study reveals that ddPCR is an alternatively potent method for the detection of EGFR T790M in the plasma samples of NSCLC patients.

BackgroundAt present, immunotherapy is a very promising treatment method for lung cancer patients, while the factors affecting response are still controversial. It is crucial to predict the efficacy of lung squamous carcinoma patients who received immunotherapy.MethodsIn our retrospective study, we enrolled lung squamous carcinoma patients who received immunotherapy at Beijing Chest Hospital from January 2017 to November 2021. All patients were grouped into two cohorts randomly, the training cohort (80% of the total) and the test cohort (20% of the total). The training cohort was used to build neural network models to assess the efficacy and outcome of immunotherapy in lung squamous carcinoma based on clinical information. The main outcome was the disease control rate (DCR), and then the secondary outcomes were objective response rate (ORR), progression-free survival (PFS), and overall survival (OS).ResultsA total of 289 patients were included in this study. The DCR model had area under the receiver operating characteristic curve (AUC) value of 0.9526 (95%CI, 0.9088–0.9879) in internal validation and 0.9491 (95%CI, 0.8704–1.0000) in external validation. The ORR model had AUC of 0.8030 (95%CI, 0.7437–0.8545) in internal validation and 0.7040 (95%CI, 0.5457–0.8379) in external validation. The PFS model had AUC of 0.8531 (95%CI, 0.8024–0.8975) in internal validation and 0.7602 (95%CI, 0.6236–0.8733) in external validation. The OS model had AUC of 0.8006 (95%CI, 0.7995–0.8017) in internal validation and 0.7382 (95%CI, 0.7366–0.7398) in external validation.ConclusionsThe neural network models show benefits in the efficacy evaluation of immunotherapy to lung squamous carcinoma patients, especially the DCR and ORR models. In our retrospective study, we found that neoadjuvant and adjuvant immunotherapy may bring greater efficacy benefits to patients.

Passion fruit wine is a popular fruit wine because of its unique aroma. However, the roles of microorganisms in different fermentation methods, particularly their contributions to aroma formation, are poorly understood. Accordingly, the goal of this study is to reveal the contribution of different fermentation methods to the flavor. Purple passion fruit was used as the experimental focus; high-throughput sequencing technology was used to analyze the microbial community of CF (controlled fermentation) and NF (natural fermentation), and the correlations between the microbial community and physicochemical indices, nonvolatile metabolites and flavor substances were analyzed. In NF, totals of eight fungal phyla, 135 fungal genera, 15 bacterial phyla and 130 bacterial genera were identified. Debaryomyces, Meyerozyma, and Wickerhamomyces were the dominant fungal genera, and Paucibacter and Pantoea were the dominant bacterial genera. In CF, totals of 11 fungal phyla, 389 fungal genera, 15 bacterial phyla and 128 bacterial genera were identified. Meyerozyma, Cladosporium, and Saccharomyces were the dominant fungal genera, and Paucibacter, Achromobacter, and Lactobacillus were the dominant bacterial genera. In NF, Wickerhamomyces, Achromobacter, Bifidobacterium and Lactobacillus were positively correlated with flavor substances such as ethylene glycol acetate formate, 2-pentanol, acetate, phenylethyl alcohol and 1-butanol, 3-methyl-. In CF, Saccharomyces, Achromobacter and Lactobacillus were positively correlated with a variety of esters and alcohols such as decanoic acid, ethyl ester, dodecanoic acid, ethyl ester and phenylethyl alcohol. Overall, this study can provide a valuable resource for further developments and improve the aromatic quality of passion fruit wine.

Sustained angiogenesis and increased PD-L1 expression on endothelial and carcinoma cells contribute toward fostering an immunosuppressive microenvironment suitable for tumor growth. PD-L1+ CTCs were reported to associate with poor prognosis in NSCLC patients. However, whether or not aneuploid circulating tumor endothelial cells (CTECs) express PD-L1, then serve as a surrogate biomarker to evaluate immunotherapy efficacy remains unknown. In this study, a novel SE-iFISH strategy was established to comprehensively quantify and characterize a full spectrum of aneuploid CTCs and CTECs in advanced NSCLC patients subjected to second-line anti-PD-1 (nivolumab) immunotherapy. In situ co-detection of diverse subtypes of aneuploid CTCs and CTECs expressing PD-L1 and Vimentin was performed. The present clinical study demonstrated that significant amounts of PD-L1+ aneuploid CTCs and CTECs could be detected in histopathologic hPD-L1- patients. In contrast to decreased PD-L1+ CTCs, the number of multiploid PD-L1+ CTECs (≥tetrasomy 8) undergoing post-therapeutic karyotype shifting increased in patients along with tumor progression following anti-PD-1 treatment. Progressive disease (PD) lung cancer patients possessing multiploid PD-L1+ CTECs had a significantly shorter PFS compared to those without PD-L1+ CTECs. In carcinoma patients, aneuploid CTCs and CTECs may exhibit a functional interplay with respect to tumor angiogenesis, progression, metastasis, and response to immunotherapy.

The rapid spread of pneumonia caused by SARS-CoV-2 has seriously threatened people. In this study, we detected the expression of anti-SARS-CoV-2 IgG/IgM and respiratory tract SARS-CoV-2 RNA in patients with COVID-19 and explored the correlation and clinical significance between SARS-CoV-2 antibody and respiratory SARS-CoV-2 RNA.From March 5, 2020 to April 28, 2020, 48 cases with COVID-19 diagnosed in Beijing Xiaotangshan Hospital were enrolled. SARS-CoV-2 RNAs were detected by real-time fluorescence RT-PCR method. Serum SARS-CoV-2 IgG/IgM antibodies were determined by colloidal gold immunochromatography. The statistical analysis was performed using chi-squared test.In all the patients, SARS-CoV-2 RNA among 270 upper respiratory tract (nasal or throat swabs) samples, 71 lower respiratory tract (sputum) samples, and anti-SARS-CoV-2 IgM/IgG antibodies in 123 serum samples were detected during the hospitalization period. The positive rate of anti-SARS-CoV-2 IgG was significantly higher than that of anti-SARS-CoV-2 IgM within the first week after symptom onset (p0.05). The positive rate of anti-SARS-CoV-2 IgG was also significantly higher than that of anti-SARS-CoV-2 IgM during day 8 - 30 after symptom onset (p0.01). The positive rate of SARS-CoV-2 RNA in the lower respiratory tract specimens (64.8%, 46/71) was significantly higher than that in the upper respiratory tract (46.7%, 126/270) (p0.05). The positive rate (100%, 4/4) of SARS-CoV-2 RNA detection in the lower respiratory tract specimens before IgG seroconversion was significantly higher than that of the positive rate (59.3%, 32/54) after IgG seroconversion (p0.01). The positive rate (72.2%, 57/79) of SARS-CoV-2 RNA detection in the upper respiratory tract specimens before IgG seroconversion was significantly higher than that of the positive rate (30.7%, 39/127) after IgG seroconversion (p0.01).Anti-SARS-CoV-2 IgG might be detected within the first week after symptom onset. The application of SARS-CoV-2 antibody (IgG/IgM) detection is important for the suspected cases of SARS-CoV-2 infection with negative SARS-CoV-2 RNA results. The positive rate of SARS-CoV-2 RNA detection in the lower respiratory tract specimens was significantly higher than that in the upper respiratory tract. Sputum detection is recommended for the detection of SARS-CoV-2 RNA. Using lower respiratory tract specimens may reduce the false negative PCR tests. The detection of SARS-CoV-2 RNA can be improved by investigating follow-up specimens over time.

With the portable and affordable gaze input devices being marketed for end users, gaze-based interactions were getting increasingly popular. Unfortunately, the understanding about the dominant task of gaze input, i.e. eye pointing task, was still not sufficient although a performance model had been specifically proposed in previous study because of that 1) the original model was based on a specific circular target condition, without the ability to predict the performance of acquiring conventional rectangular targets and that 2) there was a lack of explanation from the perspective of the anatomical structure of the eyes. In this paper, we proposed a 2D extension to take account of more general target conditions. Carrying out two experiments, we evaluated the effectiveness of the new model and furthermore we found that the index of difficulty that we redefined for 2D eye pointing (IDeye) was able to properly reflect the asymmetrical impacts of target width and height, and consequently the IDeye model could more accurately and properly predict the performance when acquiring 2D targets than Fitts’ law, no matter what kind of saccades or eye orientations (i.e. saccadic eye movement directions) was employed to acquire the desired targets. According to the results, we provided more useful implications and recommendations for gaze-based applications.

Recently, several works have studied the problem of view selection in graph databases. However, existing methods cannot fully exploit the graph properties of views, e.g., supergraph views and common subgraph views, which leads to a low view utility and duplicate view content. To address the problem, we propose an end-to-end graph view selection tool, G-View, which can judiciously generate a view set from a query workload by exploring the graph properties of candidate views and considering their efficacy. Specifically, given a graph query set and a space budget, G-View translates each query to a candidate view pattern and checks the query containment via a filtering-and-verification framework. G-View then selects the views using a graph gene algorithm (GGA), which relies on a three-phase framework that explores graph view transformations to reduce the view space and optimize the view benefit. Finally, G-View generates the extended graph views that persist all the edge-induced subgraphs to answer the subgraph and supergraph queries simultaneously. Extensive experiments on real-life and synthetic datasets demonstrated G-View achieved averagely 21x and 2x query performance speedup over two view-based methods while having 2x and 5x smaller space overhead, respectively. Moreover, the proposed selection algorithm, GGA, outperformed other selection methods in both effectiveness and efficiency.

We study a Wong–Zakai approximation for the random slow manifold of a slow–fast stochastic dynamical system. We first deduce the existence of the random slow manifold about an approximation system driven by an integrated Ornstein–Uhlenbeck process. Then, we compute the slow manifold of the approximation system, in order to gain insights of the long time dynamics of the original stochastic system. By restricting this approximation system to its slow manifold, we thus get a reduced slow random system. This reduced slow random system is used to accurately estimate a system parameter of the original system. An example is presented to illustrate this approximation.

Nivolumab, a fully human IgG4 programmed cell death-1 checkpoint inhibitor, demonstrated its benefit of prolonged survival in advanced non-small cell lung cancer (NSCLC). However, nivolumab generated unique immune-related adverse events such as thyroid dysfunctions. Herein we assessed nivolumab-induced thyroid dysfunctions in patients with previously treated advanced NSCLC in our hospital. The medical records of 11 patients with advanced NSCLC who were initiated with nivolumab treatment between June 28, 2018, and January 15, 2019, in our hospital were reviewed. Serological tests of thyroid-stimulating hormone and free tetraiodothyronine were measured at baseline and every 8 weeks. Three out of 11 patients developed new-onset hypothyroidism during the treatment, and two of three patients had transient hyperthyroidism first. Two patients were diagnosed with Grade 2 hypothyroidism and received levothyroxine therapy. The other one was Grade 1 hypothyroidism and asymptomatic. All these three patients continued nivolumab therapy. Nivolumab-induced thyroid dysfunctions in advanced NSCLC were mostly mild and controlled. Thyroid function needs to be monitored for early prediction and appropriate managements of thyroid dysfunctions.

The mechanism of regulation of PD-L1 expression by ALK translocation remains unclear. We detected PD-L1 protein expression and its regulation in lung adenocarcinoma patients with EML4-ALK fusion gene. PD-L1 and ALK expression at protein level in human lung adenocarcinoma cell lines and tumor tissue specimens was evaluated by immunohistochemistry analysis and Western blotting. The expression at DNA level and RNA level was indicated by quantitative real-time PCR analysis. The signal pathway was indicated at protein level by western blotting. The PD-L1 protein expression was higher in human lung adenocarcinoma cell lines with EML4-ALK fusion gene than that without this fusion gene. Induced expression of EML4-ALK in A549 cells significantly increased PD-L1 protein expression, whereas PD-L1 protein expression was downregulated after crizotinib and pembrolizumab successively. Significant positive correlations between PD-L1 and p-ERK, p-STAT3 or p-AKT expression were observed in ALK-translocated tumors. PD-L1 overexpression was significantly associated with shorter progressive survival and overall survival after crizotinib in ALK-translocated patients. We demonstrate that ALK translocation can upregulate PD-L1 expression by activating ERK, STAT3 and AKT pathways. ALK inhibitor combined with a PD-L1-targeted therapy may be a potential strategy in ALK-translocated lung adenocarcinoma patients.

Immune checkpoint inhibitor monotherapy is reported to have little effect in advanced non-small cell lung cancer (NSCLC) patients with driver oncogenes. However, recent studies have shown that some patients with driver genes are still benefit from combination immunotherapy after tyrosine kinase inhibitors (TKIs) drug resistance. The purpose of this study was to analyze the efficacy of posterior line immunotherapy in NSCLC patients with epidermal growth factor (EGFR) sensitive mutation, and to evaluate the value of immunotherapy in posterior line therapy in patients with advanced EGFR mutation.A total of 27 patients with EGFR mutation diagnosed in Beijing Chest Hospital, Capital Medical University from June 2018 to November 2020 were collected. After the progress of targeted therapy, they had received programmed cell death protein 1 (PD-1) checkpoint inhibitor combined with chemotherapy and anti-angiogenic drug therapy.Of the 27 advanced NSCLC patients, 19 cases (70.4%) did not have T790M mutation. There were 8 cases (29.6%) with T790M point mutation. The total objective response rate (ORR) was 40.7%. Kaplan-Meier survival analysis showed that there was no statistically significant difference among different EGFR mutations (χ²=4.15, P=0.230). But progression-free survival (PFS) was significantly longer in patients without T790M mutation than in patients with T790M mutation (9.2 mon vs 3.3 mon, χ²=2.808, P=0.041), and the same trend was observed in patients with overall survival treated with the PD-1 inhibitor (12.2 mon vs 7.3 mon, χ²=3.22, P=0.062). ORR of patients without T790M was significantly better than that with T790M (52.63% vs 12.5%, P=0.045).Patients with EGFR mutation can benefit from later-line combined immunotherapy. The patients with T790M mutation in the population of EGFR mutation had the worst effect of immunotherapy in the later line. Therefore, the follow-up treatment and whole-course management of these patients need to explore better treatment strategies to improve the benefit.【中文题目：EGFR突变晚期非小细胞肺癌患者后线 接受免疫治疗的疗效分析】 【中文摘要：背景与目的 免疫检查点抑制剂单药治疗在驱动基因阳性的晚期非小细胞肺癌（non-small cell lung cancer, NSCLC）患者中疗效甚微。研究表明，部分驱动基因阳性患者靶向治疗耐药后对免疫联合治疗仍有效。国内研究甚少。本研究旨在分析人表皮生长因子（epidermal growth factor receptor, EGFR）敏感突变NSCLC患者后线接受免疫治疗的疗效，评价真实世界免疫联合化疗在EGFR突变晚期患者后线治疗中的价值。方法 收集2018年6月-2020年11月在首都医科大学附属北京胸科医院确诊的EGFR突变的初治晚期肺腺癌患者共27例，均在靶向治疗进展后接受了程序性死亡受体1（programmed cell death protein 1, PD-1）检查点抑制剂联合化疗以及抗血管生成药物治疗。结果 27例晚期NSCLC患者中，未合并T790M突变的有19例（70.4%），合并T790M点突变的有8例（29.6%）。总客观缓解率为40.7%。Kaplan-Meier生存分析显示，不同EGFR突变类型之间接受含PD-1单抗治疗的无进展生存期（progression-free survival, PFS）均无统计学差异（χ²=4.15, P=0.23）。未合并T790M突变的患者PFS较合并T790M突变的患者显著延长（9.2个月 vs 3.3个月，χ²=2.81，P=0.041），两者总生存时间未见统计学差异（12.2个月 vs 7.3个月，χ²=3.22，P=0.062）。未合并T790M的客观缓解率明显优于合并T790M的患者（52.63% vs 12.5%, P=0.045）。结论 EGFR突变患者人群能从后线免疫联合治疗中获益，但合并T790M突变的患者后线接受免疫联合治疗疗效差。因此，这部分患者的后续治疗和全程化管理需要探索更优的治疗策略来提高获益。】 【中文关键词：肺肿瘤；表皮生长因子受体；基因突变；免疫治疗】.

The multimodal interaction technique that combines eye and foot input not only provides a great opportunity for the user with busy hands or hand disabilities to interaction with computer, but can also overcome the drawbacks when using dwell time to solve the “Midas Problem” of gaze input. However, the user’s capability of eye-foot coordination was still unclear. At the same time, the human performance in the basic task of target acquisitions by eye-foot input was also uncertain, and especially a proper performance model was lacking. Motivated by this situation, an eye pointing and foot tapping task experiment had been carried out to fill these gaps. A low-cost eye tracker and a USB foot pedal switcher were used as the input devices from different modalities. The experimental results indicated that the user was soon able to coordinate her/his foot with the eyes for target acquisitions, and that the user could respond fast to tap the foot pedal to finish a task trial in the level of 600 ms. The main performance measures of eye movement time (EMT) and eye pointing time (EPT) under the eye-foot multimodal input condition were significantly increased with the increase of saccadic amplitude A and/or the decrease of target width (size) W, and vice versa. Regression analysis shown that the \(ID_{eye}\) model was more suitable than the standard Fitts’ law to model the human performance in this multimodal interaction context.

In MOOC videos, the display positions of the subtitles are usually fixed at the bottom of the screen, which leads to the separation of the learning contents and the interpretation text (i.e. the subtitles) and thus inevitably affects the learning performance. Especially when students choose a foreign course with a non-native language as the teaching language from a global MOOC platform, this influence could be more significant. This work carried out a knowledge retention test as well as a knowledge transfer test to investigate the effects of the video subtitles with the implementation of the principle of spatial contiguity on learning performance, and further used the results of eye tracking to explain the underline mechanisms that why this principle could effectively improve the MOOC student’s performance. This paper verified the necessity of applying the principle of spatial contiguity in the subtitles of MOOC instructional videos.

This study selected a video lecture from a public MOOC platform and carried out an eye tracking experiment to investigate the effects of different representation styles on user attention. The results indicated that the subtitle positions and the background sounds in MOOC videos could significantly affect users’ attention, while same repeated scenes and different animated scenes produced using different cinematography techniques could not significantly affect users’ attention. The results of this work could benefit the designs of MOOC.

The material difference of laser additive manufacturing brake discs and traditional brake discs result in inconsistent friction coefficients of contact surfaces of the brake discs and the brake shoes during the braking process. The sliding control strategy needs to be optimized to meet the requirements of new brake discs. The model of EP2002 brake system pneumatic valve unit (PVU) is built up in simulation software of AMESim. And the correctness of the model is verified through the simulation of service brake and emergency brake. The dynamics model of train is built in MATLAB/Simulink environment, and anti-slip control logic is compiled. Considering the friction characteristics of laser additive manufacturing brake discs, anti-slip control logic was developed. To verify the validity of anti-slip logic co-simulation is conducted between AMESim and MATLAB/Simulink. The simulation results of service brake and emergency brake show that the PVU model of EP2002 reacts consistently with real systems, which verifies the correctness of the model. The simulation results of anti-slip control show that a stable performance of anti-slip is achieved even continuous slip occurred, which means that the designed anti-slip control logic meets the control requirements. The research results provide an effective model for the actual design of the train braking system and solving of failures.

This study examines the psychometric properties of a new multidimensional job insecurity measure (JIM) by O'Neill and Sevastos in a Chinese context. Overall, the results corroborate the construct validity, reliability and criterion-related validity of the JIM. Based on the results of the exploratory factor analysis, the new scale has 15 items and three items were removed from the dimensions of job loss and job change because of differences in culture and understanding between Chinese and Western employees. Additionally, the relationship between job insecurity and theoretically viable antecedents (three different types of conflicts) and outcomes (i.e., job satisfaction and counterproductive work behaviour) were also examined, and the results show that the three conflicts are effective predictors of job insecurity and job insecurity is predictor of outcomes variables. All findings show that this new JIM is more parsimonious and more effective in assessing job insecurity in the Chinese context.

HER2 mutations have emerged as oncogenic driver gene mutations in non-small cell lung cancer (NSCLC), which have not been described in detail like other driver gene mutations. Here, 295 patients with advanced lung adenocarcinoma were retrospectively screened for HER2 mutations using next-generation sequencing (NGS), and the positive cases were validated by Sanger sequencing. We identified five cases with HER2 exon 20 insertions, representing 1.7% of 295 lung adenocarcinomas. Among them, four different subtypes of HER2 exon 20 insertions were identified, including a rare subtype G778_S779insCPG never reported before with a partial response (PR) to pyrotinib and progression-free survival (PFS) of 12.8 months. Our findings reveal that HER2 exon 20 insertion mutations were detected in a small subset of lung adenocarcinomas. Given the different drug sensitivities, determining the mutation subtype by next-generation sequencing at the time of diagnosis might make sense.

Seven steady-state temperature profiles are logged in the Harbin Area, Northeast China. The temperature data illustrate high conductive thermal gradients with a mean value of 4.78°C/100 m. Temperature maps at depths of 1000, 1500, and 1800 m are represented. The present temperature field is cooling southeastward, with the highest thermal gradients being observed at the junction zone between the Central Depression and the Southeast Uplift of the Songliao Basin. The recoverable heat is calculated using a volumetric method and Monte Carlo simulation. The mean value for recoverable heat from the detailed exploration area around the Harbin Area is 1.052 × 1019 J (3.59 × 1011 t standard coal). Geological controls on the present temperature fields are discussed.

Sustained angiogenesis and increased PD-L1 expression on endothelial and carcinoma cells contribute toward fostering an immunosuppressive microenvironment suitable for tumor growth. PD-L1

Additively manufactured (AM) functionally graded porous metallic biomaterials offer unique opportunities to satisfy the contradictory design requirements of an ideal bone substitute. However, no functionally graded porous structures have ever been 3D-printed from biodegradable metals, even though biodegradability is crucial both for full tissue regeneration and for the prevention of implant-associated infections in the long term. Here, we present the first ever report on AM functionally graded biodegradable porous metallic biomaterials. We made use of a diamond unit cell for the topological design of four different types of porous structures including two functionally graded structures and two reference uniform structures. Specimens were then fabricated from pure iron powder using selective laser melting (SLM), followed by experimental and computational analyses of their permeability, dynamic biodegradation behavior, mechanical properties, and cytocompatibility. It was found that the topological design with functional gradients controlled the fluid flow, mass transport properties and biodegradation behavior of the AM porous iron specimens, as up to 4-fold variations in permeability and up to 3-fold variations in biodegradation rate were observed for the different experimental groups. After 4 weeks of in vitro biodegradation, the AM porous scaffolds lost 5–16% of their weight. This falls into the desired range of biodegradation rates for bone substitution and confirms our hypothesis that topological design could indeed accelerate the biodegradation of otherwise slowly degrading metals, like iron. Even after 4 weeks of biodegradation, the mechanical properties of the specimens (i.e., E = 0.5–2.1 GPa, σy = 8–48 MPa) remained within the range of the values reported for trabecular bone. Design-dependent cell viability did not differ from gold standard controls for up to 48 h. This study clearly shows the great potential of AM functionally graded porous iron as a bone substituting material. Moreover, we demonstrate that complex topological design permits the control of mechanical properties, degradation behavior of AM porous metallic biomaterials. Statement of Significance No functionally graded porous structures have ever been 3D-printed from biodegradable metals, even though biodegradability is crucial both for full tissue regeneration and for the prevention of implant-associated infections in the long term. Here, we present the first report on 3D-printed functionally graded biodegradable porous metallic biomaterials. Our results suggest that topological design in general, and functional gradients in particular can be used as an important tool for adjusting the biodegradation behavior of AM porous metallic biomaterials. The biodegradation rate and mass transport properties of AM porous iron can be increased while maintaining the bone-mimicking mechanical properties of these biomaterials. The observations reported here underline the importance of proper topological design in the development of AM porous biodegradable metals.

Background To explore the efficacy and safety of crizotinib versus platinum-based double agent chemotherapy as the first-line treatment in patients with advanced anaplastic lymphoma kinase (ALK)-positive lung adenocarcinoma. Method We retrospectively analyzed data from 19 patients with advanced ALK-positive lung adenocarcinoma who had received no previous systemic treatment for advanced disease. Seven patients received oral crizotinib at a dose of 250 mg twice daily; 12 patients were administered standard chemotherapy (pemetrexed, paclitaxel, vinorelbine or gemcitabine plus either cisplatin or carboplatin) every three weeks for up to six cycles. The primary endpoint was overall response rate (ORR), disease control rate (DCR), and safety. Results The ORR was significantly higher with crizotinib than with chemotherapy (83.3% in the crizotinib vs. 25.0% in the chemotherapy group, P < 0.05); the DCRs were 100% and 75%, respectively (P < 0.05). The common adverse events associated with crizotinib were visual abnormality and diarrhea, whereas those associated with chemotherapy were neutropenia and nausea. In the crizotinib group, liver aminotransferase elevation (adverse events grade 3 or 4) occurred in one patient (14.3%). In the chemotherapy group, the same grade neutropenia adverse event occurred in two patients (16.6%). The incidence of treatment-related grade 3 or 4 adverse events was similar in both groups. Compared with chemotherapy, crizotinib was associated with a greater reduction in lung cancer symptoms and a greater improvement in quality of life. Conclusion As a first-line treatment, crizotinib was superior to platinum-based double chemotherapy in patients with previously untreated advanced ALK-positive lung adenocarcinoma. Therefore, crizotinib is an optimal therapy as a first-line treatment in these patients.

Non-small cell lung cancer (NSCLC) has been transformed from the treatment according to histological type to genotype treatment model. The epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK) genes are the most important drivers in lung cancer. The aim of this study is to explore the clinical characteristics and prognostic factors of patients with advanced NSCLC with different genotypes.We retrospectively reviewed the clinical data of 553 advanced NSCLC patients with EGFR mutations and ALK positive who were hospitalized in the Beijing Chest Hospital from July 2004 to December 2015, and the independent prognostic factors of patients were analyzed by Cox proportional hazards regression model.The clinical data of 553 patients (227 with EGFR mutations, 58 with ALK positive, 2 with EGFR and ALK co-mutation and 266 with wild-type) with advanced NSCLC were enrolled in this study. The median survival time of 227 patients with EGFR mutations was 28.7 mo (95%CI: 22.160-35.240), and the performance status (PS) score (0-1) (HR=4.451; 95%CI: 2.112-9.382; P0.001) and EGFR-tyrosine kinase inhibitors (TKIs) targeted therapy (HR=2.785; 95%CI: 1.871-4.145; P0.001) were the independent prognostic factors for the survival of patients harboring EGFR mutations. The median survival time of 58 patients with ALK positive was 15.5 mo (95%CI: 10.991-20.009), and treatment with crizotinib (P=0.022) was the independent influence factor for the survival of ALK positive patients. The median survival time of 266 patients with wild-type was 12.1 mo (95%CI: 10.660-13.540), and the PS score (0-1) (HR=2.313; 95%CI: 1.380-3.877; P=0.001) and treatment with chemotherapy (HR=1.911; 95%CI: 1.396-2.616; P0.001) were the independent prognostic factors for the survival of wild-type patients CONCLUSIONS: The prognosis of patients with advanced NSCLC is associated with genetic mutation, and targeted therapy has a improvement on survival for patients with EGFR mutations or ALK rearrangement.背景与目的 非小细胞肺癌（non-small cell lung cancer, NSCLC）已由原来的组织分型指导下的治疗转变为基因分型指导治疗的模式，表皮生长因子受体（epidermal growth factor receptor, EGFR）和间变性淋巴瘤激酶（anaplastic lymphoma kinase, ALK）是肺癌最重要的两个驱动基因。本研究旨在探讨不同基因分型的复发或转移晚期NSCLC患者的临床特点及预后影响因素。方法 回顾性分析北京胸科医院2004年7月-2015年12月间553例EGFR和ALK基因状态明确的晚期NSCLC患者的临床资料，采用Cox比例风险回归模型对患者预后的独立影响因素进行分析。结果 553例细胞学或组织学证实的晚期NSCLC患者，EGFR突变患者227例，ALK阳性患者58例，EGFR和ALK双突变患者2例，EGFR和ALK野生型患者266例。227例EGFR突变患者的中位生存期（overall survival, OS）为28.7个月（95%CI: 22.160-35.240），体能状态（performance status, PS）评分为0分-1分（HR=4.451; 95%CI: 2.112-9.382; P0.001）、接受EGFR-酪氨酸激酶抑制剂（tyrosine kinase inhibitors, TKIs）靶向治疗（HR=2.785; 95%CI: 1.871-4.145; P0.001）是EGFR突变患者生存的独立影响因素。58例ALK阳性患者的中位OS为15.5个月（95%CI: 10.991-20.009），接受克唑替尼靶向治疗（P=0.022）是ALK阳性患者生存的独立影响因素。266例野生型患者的中位OS为12.1个月（95%CI: 10.660-13.540），PS评分为0分-1分（HR=2.313; 95%CI: 1.380-3.877; P=0.001）、接受化疗（HR=1.911; 95%CI: 1.396-2.616; P0.001）是野生型患者生存的独立影响因素。结论 不同基因型的晚期NSCLC患者的预后差异较大，靶向治疗可改善EGFR突变、ALK阳性患者生存。.

The success of cell therapy for tissue repair and regeneration demands efficient and reliable cell delivery methods. Here we established a novel microengineered cryogel (microcryogel) array chip containing microcryogels with predefined size and shape as injectable cell delivery vehicles. The microscale macroporous cryogels enabled automatic and homogeneous loading of tailored cellular niches (e.g. cells, matrices, bioactive factors) and could be easily harvested from the ready-to-use array chip. In contrast to microscale hydrogels, microcryogels exhibited excellent elasticity and could retain their shape and integrity after injection through the microsyringe routinely used for cell therapy. Human mesenchymal stromal cells loaded within microcryogels could be shielded from the mechanical insult and necrosis caused by during direct cell injection. After subcutaneous injection to the mice, cell-loaded microcryogels exhibited concentrated localization and enhanced retention at the injection site compared to dissociated cells. To demonstrate the potential therapeutic application for ischemic diseases, site-directed induction of angiogenesis was achieved subcutaneously in mice 2weeks after injection of NIH/3T3 fibroblast-loaded microcryogels, indicating long-term engraftment, accumulative paracrine stimulation and augmented host tissue integration. Our results convincingly showed the great promise of microcryogels as 3-D cellular microniches and injectable cell delivery vehicles to tackle major challenges faced by cell therapy-based regenerative medicine including shear-induced damages, uncontrolled localization, poor retention, limited cellular survival and functionalities in vivo.

Cell-based high throughput drug screening accelerates the pace of drug discovery which is routinely operated on planar high-density multi-well plates with sophisticated robotic liquid-dispensing systems for cell seeding and drug administration. Considerable efforts have been made to upgrade in vitro cellular models from 2D to a more biomimetic 3D configuration. For instance, in anti-cancer drug screening, tumor spheroids are increasingly applied as a gold-standard 3D model exhibiting cellular behaviors and drug responses distinguishable from the 2D counterpart. However, translation of spheroids to high throughput drug screening is challenging since pre-formation of spheroids and subsequent translocation to multi-well plates for drug testing are usually uncontrollable and time/reagent consuming and cell loss is inevitable during medium exchange for drug testing. Here we present an off-the-shelf micro-scaffold array chip which enables high throughput 3D cell culture, drug administration and quantitative in situ assays entirely on the same chip. The sponge-like micro-scaffolds functioned both as absorbents to realize parallel auto-loading of cells or drugs and as barriers to prevent cell loss during medium exchange via centrifugation. Rapid manual loading of cell suspensions or drugs into the 96 isolated micro-scaffolds on the chip was achieved in the timescale of several seconds, meanwhile with total medium consumption reduced to the order of microliters. Proof of concept demonstration of drug cytotoxicity testing was performed on multiple cancer cells using common benchtop equipment, making it accessible to most biomedical labs with basic cell culture setups. Higher cellular drug resistance was constantly obtained with this platform compared to the planar cultures, which was partially attributed to the malignant phenotype of cancer cells yielded by enhanced cell-matrix interactions in the micro-scaffolds. Interestingly, the high drug resistance of 3D cultured cells in the micro-scaffold was shown to be density-independent in contrast to the density-dependent drug response for 2D cultured cells, indicating intrinsic differences between the two culture models. This platform is expected to facilitate upgrade of the current cell-based high throughput drug testing to the 3D level and be widely applicable across various disciplines.

Somatic DNA mutations affecting the epidermal growth factor receptor (EGFR) signaling pathway are known to predict responsiveness to EGFR-tyrosine kinase inhibitor drugs in patients with advanced non-small-cell lung cancers. We evaluated a sensitive liquidchip platform for detecting EGFR, KRAS (alias Ki-ras), proto-oncogene B-Raf, and phosphatidylinositol 3-kinase CA mutations in plasma samples, which were highly correlated with matched tumor tissues from 86 patients with advanced non-small-cell lung cancers. Either EGFR exon 19 or 21 mutations were detected in 36 patients: 23 of whom had identical mutations in both their blood and tissue samples; whereas mutations in the remaining 13 were found only in their tumor samples. These EGFR mutations occurred at a significantly higher frequency in females, never-smokers, and in patients with adenocarcinomas (P ≤ 0.001). The EGFR exon 20 T790M mutation was detected in only one of the paired samples [100% (95% CI, 96% to 100%) agreement]. For KRAS, proto-oncogene B-Raf, and phosphatidylinositol 3-kinase CA mutations, the overall agreements were 97% (95% CI, 90% to 99%), 98% (95% CI, 92% to 99%), and 97% (95% CI, 90% to 99%), respectively, and these were not associated with age, sex, smoking history, or histopathologic type. In conclusion, mutations detected in plasma correlated strongly with mutation profiles in each respective tumor sample, suggesting that this liquidchip platform may offer a rapid and noninvasive method for predicting tumor responsiveness to EGFR-tyrosine kinase inhibitor drugs in patients with advanced non-small-cell lung cancers.

There is no high-level evidence for the time of whole brain radiotherapy (WBRT) for patients with epidermal growth factor receptor (EGFR)-mutated non-small cell lung cancer (NSCLC) and brain metastases. The aim of this study is to assess the appropriate timing of WBRT for patients with EGFR-mutated NSCLC and brain metastases (BM).There were 78 patients diagnosed with EGFR-mutated NSCLC and BM in Beijing Chest Hospital between August 2009 and May 2015. 48 untreated patients who received both WBRT and EGFR-tyrosine kinase inhibitors (TKIs) therapy. Prognostic factors of intracranial progression-free survival (PFS) and overall survival (OS) were identified by Cox proportional hazards modeling.Intracranial objective response rate was 81.3% and disease control rate was 93.8%. Median intracranial PFS was 10 months. Median OS was 18 months. Multivariate analysis of intracranial PFS revealed that Eastern Cooperative Oncology Group (ECOG) performance status (PS) 0-1 (HR=30.436, 95%CI: 4.721-196.211, P0.001) and early WBRT (HR=3.663, 95%CI: 1.657-8.098, P=0.001) had a better intracranial PFS. Multivariate analysis of OS revealed that PS 0-1 (HR=57.607, 95%CI: 6.135-540.953, P0.001), early WBRT (HR=2.757, 95%CI: 1.140-6.669, P=0.024), and stereotactic radiosurgery (HR=5.964, 95%CI: 1.895-18.767, P=0.002) were independent prognostic factors of OS.Early WBRT combined with EGFR-TKIs can improve outcomes of patients with EGFR-mutated NSCLC and BM, but it needs to be confirmed by large-sample-size and multicenter prospective clinical trials.背景与目的 全脑放疗（whole brain radiotherapy, WBRT）在表皮生长因子受体（epidermal growth factor receptor, EGFR）突变的非小细胞肺癌（non-small cell lung cancer, NSCLC）脑转移患者治疗中何时应用尚无高级别的循证医学证据。本研究旨在探讨WBRT的参与时间对携有EGFR突变的NSCLC脑转移患者生存的影响。方法 2009年8月-2015年5月在我院确诊的EGFR突变伴脑转移的晚期NSCLC共78例患者，均接受WBRT及EGFR酪氨酸激酶抑制剂（EGFR tyrosine kinase inhibitors, EGFR-TKIs）治疗的48例初治患者进入临床分析，采用Cox比例风险模型分析患者颅内无进展生存期（progression-free survival, PFS）及总生存期（overall survival, OS）的影响因素。结果 全组患者颅内客观缓解率（objective response rate, ORR）为81.3%，颅内疾病控制率（disease control rate, DCR）为93.8%，中位颅内PFS为10个月，中位OS为18个月。颅内PFS的多因素分析显示，美国东部肿瘤协作组评分（Eastern Cooperative Oncology Group performance status, ECOG PS）0分-1分（HR=30.436, 95%CI: 4.721-196.211, P0.001）及早期WBRT患者（HR=3.663, 95%CI: 1.657-8.098, P=0.001）的颅内PFS更佳。OS的多因素分析显示，ECOG PS 0分-1分（HR=57.607, 95%CI: 6.135-540.953, P0.001）、早期WBRT（HR=2.757, 95%CI: 1.140-6.669, P=0.024）及立体定向放射外科（stereotaxic radio surgery, SRS）的应用（HR=5.964, 95%CI: 1.895-18.767, P=0.002）是患者OS的独立预后因素。结论 早期WBRT联合TKIs治疗可改善EGFR突变的NSCLC脑转移患者的预后，尚有待大样本的前瞻性临床试验验证。.

This paper is devoted to studying parameter estimation for a class of stochastic dynamical systems with oscillating coefficients. We show that the homogenized systems faithfully capture the dynamical quantities such as mean exit time and escape probability. Exacting data from observations on the mean exit time (or escape probability) of the original systems, we try to fit the mean exit time (or escape probability) of the homogenized systems by least square method. In this way, we can accurately estimate the unknown parameter in the drift under appropriate assumptions. Furthermore, we conduct some numerical experiments to illustrate our method.

Icotinib is the first self-developed small molecular drug in China for targeted therapy of lung cancer. Compared to the other two commercially available epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors, gefitinib and erlotinib, icotinib is similar to them in chemical structure, mechanism of activity and therapeutic effects. To explore the efficacy and side effects of icotinib hydrochloride in the treatment of the advanced non-small cell lung cancer (NSCLC) patients with EGFR mutation and wild-type.Patients with advanced NSCLC who were treated with icotinib hydrochloride in Beijing Chest Hospital were retrospective analyzed from March 2009 to December 2014.The clinical data of 124 patients (99 with EGFR mutation and 25 with wild type) with advanced NSCLC were enrolled in this study. The patients' overall objective response rate (ORR) was 51.6 % and the disease control rate (DCR) was 79.8%; The patients with EGFR mutation, ORR was 63.6%, DCR was 93.9%. The ORR was 4.0% and the DCR was 24.0% in the wild-type patients. Median progression-free survival (PFS) with icotinib treatment in EGFR mutation patients was 10.5 months and 1.0 month in wild-type patients. The major adverse events were mild skin rash (30.6%) and diarrhea (16.1%).Monotherapy with icotinib hydrochloride is effective and tolerable for the advanced NSCLC EGFR mutation patients. 背景与目的 盐酸埃克替尼（icotinib hydrochloride）是我国第一个具有自主知识产权的小分子靶向抗癌新药，与吉非替尼和厄洛替尼相比，在化学结构、分子作用机理、疗效等方面相似。本研究观察盐酸埃克替尼治疗表皮生长因子受体突变状态明确的晚期非小细胞肺癌（non-small cell lung cancer, NSCLC）的疗效和毒副反应。方法 回顾性分析2009年3月-2014年12月间北京胸科医院收治的晚期NSCLC患者，表皮生长因子受体（epidermal growth factor receptor, EGFR）突变状态已知，均口服盐酸埃克替尼治疗，评价其疗效和毒副反应。结果 124例组织学证实的晚期NSCLC患者，其中EGFR突变型99例，野生型25例。全组客观有效率（objective response rate, ORR）为51.6%，疾病控制率（disease control rate, DCR）为79.8%。突变型和野生型患者的ORR：63.6% vs 4.0%，DCR：93.9% vs 24.0%，两者均有统计学差异（P0.000,1）。突变型和野生型患者的无进展生存期（progression-free survival, PFS）（分别为10.5个月和1.0个月）（P0.000,1）。治疗相关的毒副反应主要为皮疹38例（30.6%），腹泻20例（16.1%）。结论 盐酸埃克替尼治疗EGFR突变的晚期NSCLC疗效肯定，耐受性好。.

Activating mutations in epidermal growth factor receptor (EGFR) and KRAS are important markers in non-small cell lung cancer. However, EGFR and KRAS gene mutations in lung squamous cell carcinoma are rarely reported. The aim of this study was to analyze EGFR and KRAS gene mutation rate and their relationship with clinical features in patients with lung squamous cell carcinomas.A total of 139 patients undergoing treatment for naïve lung squamous cell carcinomas with tumor tissue samples available for testing were recruited. EGFR and KRAS mutation statuses of the tumor samples were detected using a mutant enriched liquid chip.Of the 139 cases of lung squamous cell carcinoma, EGFR mutations were detected in 25 cases (18%), KRAS mutations were detected in 7 cases (5%), and the presence of both EGFR and KRAS mutations was detected in 1 case (0.7%). EGFR mutations occurred more often in females than in males (33.3% vs 16.5%) and in patients that never smoked than in those who smoke (29.6% vs 16.1%). However, the difference did not reach statistical significance (P0.05). No significant differences were observed in age, stage, and different biopsy type. KRAS mutations occurred more often in males than in females (5.5% vs 0%), but the difference did not reach statistical significance (P0.05). No significant differences were observed in age, stage, different biopsy type, and smoking status (P0.05).EGFR and KRAS mutations were low in lung squamous cell carcinomas, and had no significant correlation with clinical features. Before using tyrosine kinase inhibitor targeted therapy, EGFR and KRAS mutations should be detected in patients with lung squamous cell carcinomas.背景与目的 表皮生长因子受体（epidermal growth factor receptor, EGFR）突变和KRAS基因突变是非小细胞肺癌（non-small cell lung cancer, NSCLC）靶向治疗的重要分子标志，但关于肺鳞癌中EGFR和KRAS基因突变情况的报道甚少。本研究旨在分析肺鳞癌EGFR和KRAS基因突变与临床特征的关系。方法 收集初治肺鳞癌患者139例，有可供检测的肿瘤组织标本。利用突变富集液相芯片法进行EGFR和KRAS基因突变检测。结果 139例肺鳞癌中，EGFR基因突变25例（18%），KRAS基因突变7例（5%），EGFR和KRAS基因同时发生突变1例（0.7%）。女性和不吸烟患者EGFR基因突变率高于男性和吸烟患者（33.3% vs 16.5%, 29.6% vs 16.1%），但差异均无统计学意义（P0.05, P0.05）；不同年龄、分期及病理取材标本之间差异均无统计学意义（P0.05）。男性患者KRAS基因突变率高于女性患者（5.5% vs 0%），但差异无统计学意义（P0.05）；在不同年龄、分期、病理取材标本及是否吸烟各亚组分析中KRAS基因突变差异无统计学意义（P0.05）。结论 肺鳞癌患者EGFR和KRAS基因突变发生率均较低，且都与临床特征无明显相关。肺鳞癌患者使用酪氨酸激酶抑制剂（tyrosine kinase inhibitors, TKIs）靶向治疗药物之前，也应检测EGFR基因和KRAS基因的突变情况。.

Status of epidermal growth factor receptor (EGFR) gene is a predictor of response to EGFR tyrosine kinase inhibitor (TKI). However, little is know about the relationship between EGFR status and response to chemotherapy. We evaluated the prediction value of EGFR mutation status on response to first-line chemotherapy in patients with advanced non-small-cell lung cancer (NSCLC).The data of 181 patients with stage IIIb/IV NSCLC who diagnosed by histopathology from January 10, 2006 to December 20, 2013 in Beijing Chest Hospital, Capital Medical University were collected. The relationships between EGFR gene status, clinical characteristics and response and progression-free survival (PFS) were analyzed.All of the 181 patients' EGFR statuses were determined. 75 (41.4%) patients harbored EGFR-activating mutations and 106 (58.6%) patients were EGFR wild-type. All patients received first-line chemotherapy. The objective response rate (ORR) was 26.0% and disease control rate (DCR) was 70.2%. Patients with EGFR-activating mutations had a higher DCR than patients with EGFR wild-type (84.0% vs 60.4%, P=0.001) did. Subgroup analysis showed that the ORR and DCR in patients with EGFR exon 19 deletions were remarkably higher than those with EGFR wild-type (P = 0.049, 0.002, respectively). The DCR in patients with EGFR exon 21 L858R mutation was significantly higher than that in patients with EGFR wild-type (P=0.010). 168 patients were available for response evaluation in all of 181 patients and median PFS was 4.3 mo. The PFS of patients with adenocarcinoma was significantly higher than that patients with squamous cell carcinoma (4.7 mo vs 3.0 mo, P=0.036). The PFS in patients harbored EGFR-activating mutations was significantly higher than that in the patients with EGFR wild-type (6.3 mo vs 3.0 mo, P=0.002). The PFS of patients with a performance status (PS) of 0-1 was significantly higher than that in patients with a PS of 2 (4.4 months vs. 0.7 months, P= 0.016). Cox multivariate analysis indicates the EGFR-activating mutation is an independent factor affecting PFS (HR=0.654, 95%CI: 0.470-0.909, P=0.012).EGFR-activating mutation is a predictor for PFS of first-line chemotherapy in advanced NSCLC patients. .背景与目的 表皮生长因子受体（epidermal growth factor receptor, EGFR）基因状态是表皮生长因子受体酪氨酸激酶抑制剂（epidermal growth factor receptor tyrosine kinase inhibitor, EGFR-TKI）疗效的预测因素，但其对化疗疗效的预测作用尚不明确。本研究旨在探讨对晚期非小细胞肺癌（non-small cell lung cancer, NSCLC）患者一线化疗疗效的预测意义。方法 收集首都医科大学附属北京胸科医院自2006年1月10日-2013年12月20日经组织病理学证实的181例IIIb期/IV期的NSCLC患者。分析EGFR基因状态、临床特征与化疗疗效及无疾病进展生存期（progression-free survival, PFS）之间的关系。结果 181例患者均进行了EGFR基因检测，EGFR突变患者75例（41.4%），野生型为106例（58.6%）。全部患者均接受一线化疗，客观缓解率（objective response rate, ORR）为26.0%，疾病控制率（disease control rate, DCR）为70.2%。EGFR突变患者的DCR显著高于EGFR野生型患者高（84.0% vs 60.4%, P=0.001）。亚组分析显示，19外显子缺失突变患者化疗的ORR、DCR均高于EGFR野生型患者（P值分别为0.049，0.002）。21外显子L858R突变患者的DCR高于EGFR野生型患者（P=0.010）。全部患者中，168例患者可评价PFS，中位PFS为4.3个月，其中腺癌患者PFS较鳞癌患者延长（4.7个月 vs 3.0个月，P=0.036）；突变患者PFS长于野生型患者（6.3个月 vs 3.0个月，P=0.002）；体力状况评分（performance status, PS）0-1分组患者PFS较评分为2分延长（4.4个月 vs 0.7个月，P=0.016）。Cox多因素分析显示，EGFR突变是影响PFS的独立因素（HR=0.654, 95%CI: 0.470-0.909, P=0.012）。结论 EGFR突变是晚期NSCLC患者一线化疗PFS的预测因素。.

The alphavirus Semliki Forest virus (SFV) infects cells via a low-pH-dependent membrane fusion reaction mediated by the E1 envelope protein. Fusion is regulated by the interaction of E1 with the receptor-binding protein E2. E2 is synthesized as a precursor termed “p62,” which forms a stable heterodimer with E1 and is processed late in the secretory pathway by a cellular furin-like protease. Once processing to E2 occurs, the E1/E2 heterodimer is destabilized so that it is more readily dissociated by exposure to low pH, allowing fusion and infection. We have used FD11 cells, a furin-deficient CHO cell line, to characterize the processing of p62 and its role in the control of virus fusion and infection. p62 was not cleaved in FD11 cells and cleavage was restored in FD11 cell transfectants expressing human furin. Studies of unprocessed virus produced in FD11 cells (wt/p62) demonstrated that the p62 protein was efficiently cleaved by purified furin in vitro, without requiring prior exposure to low pH. wt/p62 virus particles were also processed during their endocytic uptake in furin-containing cells, resulting in more efficient virus infection. wt/p62 virus was compared with mutant L, in which p62 cleavage was blocked by mutation of the furin-recognition motif. wt/p62 and mutant L had similar fusion properties, requiring a much lower pH than control virus to trigger fusion and fusogenic E1 conformational changes. However, the in vivo infectivity of mutant L was more strongly inhibited than that of wt/p62, due to additional effects of the mutation on virus-cell binding.

This work is about low dimensional reduction for a slow-fast data assimilation system with non-Gaussian stable Levy noise via stochastic averaging. When the observations are only available for slow components, we show that the averaged, low dimensional filter approximates the original filter, by examining the corresponding Zakai stochastic partial differential equations. Furthermore, we demonstrate that the low dimensional slow system approximates the slow dynamics of the original system, by examining parameter estimation and most probable paths.

Graph cut based interactive image segmentation attracts much attention in recent years. Given an image, traditional methods generally need users to indicate parts of foreground and background by drawing strokes, etc. Next, these methods formulate energy functions, which are generally composed of color and gradient constraints. Considering that many objects to be cut out are compact, the paper presents a method that incorporates a simple but effective direct connectivity constraint. The constraint is defined geometrically based on the user input strokes. The centers of those foreground strokes are treated as foreground representing points. Pixels to be labeled, which are not directly connected to the representing points, i.e. blocked by the background strokes from the representing points, are considered to belong to the background. Results show that with the same amount of user interaction, the proposed segmentation method obtains better results than state-of-the-art ones.

Graph cut based interactive segmentation is useful to extract objects from images. Color and gradient constraints are two terms appearing in most of energy functions of related methods. In order to balance the two constraints, state-of-the-art methods adopt a pre-given fixed weight. However, different images and even different parts in a single image have different demands for proportion of the two constraints. This paper proposes a graph cut based segmentation method which is capable of intelligently balancing the two constraints on the fly. Particularly, it analyzes the demand of each pixel for color and gradient constraints and arranges a weight at the pixel to balance the two, automatically. Results show that the proposed method obtains better results than traditional ones.

Zhang et al. (2010) proposed a performance model for dwell-based eye pointing. However, their model was based on a specific circular target condition, without the ability to predict the performance of acquiring conventional rectangular targets. In this paper, we extend their one-dimensional model to two-dimensional (2D) target conditions. Carrying out an experiment, we evaluate the abilities of different model candidates to find out the most appropriate one. The new index of difficulty we redefine for 2D eye pointing (IDeye) can properly reflect the asymmetrical impact of target width and height, and consequently the IDeye model can accurately predict the performance for 2D targets (R2 > 0.9). According to the results of our study, the new model can provide more useful design implications for gaze-based interactions.

Fitts' law is the most widely applied model in the field of HCI. However, this model and its existing extensions are still limited for 2D pointing task especially when the effects of movement direction (Θ) remain in the task. In this paper, we employ the concept of projection to account for the effects of target width (W) and height (H) on movement time so that we seamlessly integrate the four factors, i.e. Θ, amplitude (A), W and H, into the new extension of Fitts' law, which can uncover not only the periodicity of the asymmetrical impacts of W and H with the variation of Θ but also their interrelation. Carrying out two experiments, we verify that the vertical projection of W and the horizontal projection of H in the line of movement direction can be viewed as the determinants of movement time. Finally, we offer recommendations for 2D pointing experiments and discuss the implications for interface designs.

In this paper, we present a target searching experiment to investigate how long is long enough to maintain the speed-accuracy trade-off in eye pointing tasks that use dwell time as the activation mechanism. The experimental task, which took account of three factors including cognitive complexity, dwell time and visual feedback mode, mixes visual search and target acquisition together. In other words, the subjects need to search for and recognize the target before the final selection in each trial. The results clarify the suitable ranges of dwell time for users to avoid wrong selections as possible as they can under different cognitive load conditions. We also discussed the implications for user interface designs.

To obtain a recombinant human plasminogen (hPLG) with potential anti-platelet aggregation activity, we cloned the cDNA coding Pro544 to Asn791 of hPLG, a kringle-deficit derivative (hPLG-deltaK). The Pro559 in activation loop was then mutated into Asp559 to provide Arg-Gly-Asp (RGD) motif. The constructed pPICZalphaA-RGD-HPLG-deltaK plasmid was expressed in yeast Pichia pastoris GS115, which produced RGD-hPLG-deltaK about 0.160 g/L broth. After affinity chromatography, the purity of the recombinant protein reached above 90%. Western blotting test confirmed that it retained the immunological reaction capability as human PLG. Its urokinase activation rate in 24 hours and its fibrinolytic activity made no deference against native hPLG-deltaK (P=0.630, n=5). Importantly, after activation by urokinase, RGD-hPLG-deltaK showed a significantly higher platelet aggregation inhibition rate (Ri) (21.8% +/- 1.57%) than hPLG-deltaK (3.8% +/- 0.33%) (P=0.000, n=5). These results proved that we constructed an hPLG mutant with anti-platelet aggregation activity, which made a foundation for developing innovative thrombolytic drugs with multifunction.

lag-2 encodes a ligand for LIN-12/Notch and is a component of the lateral signal that activates LIN-12/Notch during Caenorhabditis elegans vulval precursor cell (VPC) fate patterning. lag-2 is specifically transcribed in one VPC, named P6.p, in response to activation of EGFR/Ras/MAPK by the inductive signal that initiates vulval development. Here, we show that a critical molecular event linking inductive and lateral signaling is the relief of VPC-wide lag-2 repression in P6.p. We find that the lag-2 promoter contains an element, VPCrep, which mediates repression in all VPCs when the inductive signal is absent, and another promoter element, VPCact, which is required for activation when repression is relieved by the inductive signal. We show that repression through VPCrep is mediated by the Elk1 ortholog LIN-1, and that the level and subcellular accumulation of a functional LIN-1::GFP protein is similar in all six VPCs before and after vulval induction, suggesting that relief of LIN-1–mediated repression in P6.p is likely due to the known MAPK-dependent phosphorylation of LIN-1. We also provide evidence that the factor(s) acting through VPCact is present in all VPCs but is not modulated by the inductive signal, and that transcription of lag-2 requires the Hth/Meis ortholog UNC-62 and the Mediator complex component SUR-2. Relief of repression of lag-2 in P6.p offers a plausible mechanistic basis for spatial restriction of lag-2 in generating the precise spatial pattern of VPC fates.

To evaluate the efficacy, side-effects, median survival duration and survival rate of vinorelbine (NVB) combined with cisplatin (DDP) in the treatment of non-small cell lung cancer (NSCLC).A total of 220 patients with inoperable NSCLC received NVB and DDP combined chemotherapy: NVB 25-30 mg/(m²*d) on days 1 and 5 (or 8), DDP 60-80 mg/(m²*d) on day 2. The schedule was repeated every 28 days. The efficacy and side-effects were analysed and followed-up after at least two cycles of chemotherapy.The overall response rate (CR+PR) was 30.9% (68/220). The response rate was 31.3% (51/163) in initial treatment group, and 29.8% (17/57) in retreatment group. The median survival duration was 8.3 months. The 1-, 2- and 3-year survival rates were 39.23%, 19.31% and 6.32%, respectively. The main side-effects were myelosuppression and digestive tract reactions.Vinorelbine plus cisplatin is an effective and well-tolerated regimen for non small cell lung cancer and myelosuppression is its dose-limiting toxicity.

In this paper, we present an attention task experiment that investigated the effects of different visual feedback forms on the eye cursor's stability to find out the well-formed visual feedback. The different feedback forms were designed for dwell time, the eye cursor, and the center of the target (marked as a focus area). Our experimental findings can provide useful implications for the design of eye-controlled interfaces.

Uroacitides is a group of cell differentiation inducers, which is purified from fresh human urine. Preclinical studies of Uroacitides have showed that cancer cells could be induced to differentiate, and the growth of cancer cells could be inhibited by Uroacitides. The aim of this study is to compare the efficacy and toxicity between Uroacitides combined with NP regimen and NP alone in treatment of advanced non-small cell lung cancer (NSCLC).Forty-two cases of advanced NSCLC were randomized into Uroacitides+NP and NP groups. NP group: NVB 25mg/m² on days 1 and 8, DDP 75mg/m² on day 1. Uroacitides combined with NP group: Uroacitides of 300mL was given through subclavian catheter daily for 7 days prior to the NP chemotherapy, then concurrently with NP regimen for 2 cycles, except the days of administration of chemotherapy.In the Uroacitides+NP group, the overall response rate was 44.4%, and 20.0% in the NP group (P0.05). The median survival time was 9 months in the Uroacitides+NP group and 6 months in the NP group (P=0.0287). The main toxicities were myelosuppression, gastrointestinal reaction and alopecia, and there was no significant difference in incidences of toxicities between the two groups (P0.05).Uroacitides combined with NP regimen shows a good curative effect and low toxicity, and may significantly prolong the median survival time for advanced NSCLC.

Matrix metalloproteinase-9 (MMP-9), endostatin (ES) and vascular endothelial growth factor (VEGF) are important angiogenic regulators for many neoplasms. The aim of this study is to judge clinical and prognostic values of detection of serum MMP-9, ES and VEGF in patients with non-small cell lung cancer (NSCLC).Serum levels of MMP-9, ES and VEGF were detected in 92 patients with NSCLC, 50 patients with pulmonary benign disease and 52 healthy controls by ELISA method.The serum levels of MMP-9, ES and VEGF in NSCLC patients were significantly higher than those in patients with pulmonary benign disease and healthy controls (P=0.000, P=0.000, P=0.000). The sensitivity and specificity of serum MMP-9 was 92.51% and 79.10% with a cutoff value of 117.17 μg/L, 88.32% and 74.25% for ES with a cutoff value of 100.31 μg/L, and 83.40% and 75.63% for VEGF with a cutoff value of 380.32 ng/L. Serum MMP-9 and ES levels were significant prognostic factors for lung cancer patients (P=0.0145, P=0.008). The change of serum MMP-9 level after chemotherapy was a useful indicator of prognosis for NSCLC patients (P=0.0322).The serum levels of MMP-9, ES and VEGF are significantly increased in patients with NSCLC. They might be used as prognostic parameters in patients with NSCLC.

It has been proven that epigermal growth factor receptor (EGFR) signal pathway plaied an important role in the oncogenesis and development of non-small cell lung cancer (NSCLC). EGFR tyrosine kinase inhibitors (EGFR-TKIs) are currently investigated in the treatment of NSCLC. It was suggested in previous studies that the EGFR gene mutations were correlated with the response to EGFR-TKIs therapy and prognosis of NSCLC. We studied the role of EGFR gene mutations in response to two kinds of TKIs therapy and prognosis of NSCLC in this study.The tissue samples of 59 advanced NSCLC patients (34 patients receiving Gefitinib monotherapy and 25 patients receiving Erlotinib monotherapy) were collected, and patient charts were reviewed. The mutations in exons 19 and 21 of EGFR gene were detected by PCR-PAGE and PCR-RFLP respectively. The sequences of interested fragments were verified by direct sequencing. Relationship between EGFR mutation and response to TKIs therapy was analyzed with Chi-Square test.EGFR gene mutations were identified in 22 of 59 samples (37.3%). EGFR gene mutation rate was significantly higher in female, non-smoker and patients with adenocarcinoma than in others (50% vs 18.9%, P0.05). The patients with EGFR gene mutation had a better response to TKIs therapy than those without (86.4% vs 54.1%,P0.05). The patients with EGFR gene mutation had slower disease progression and longer overall survival than those without, but statistically non-significant (P0.05).EGFR gene mutation occurs more frequently in female, non-smoker and patients with adenocarcinoma. In patients with advanced NSCLC, EGFR mutation is associated with good response to EGFR-TKIs therapy.

Erlotinib is a small molecular inhibitor of tyrosine kinase. Multiple foreign and demestic studies have confirmed that it can prolong the median progression-free survival time (PFS) and the overall survival (OS), which could be more significant in the selected population. In this study we retrospectively oberserved the response, the survival and adverse reaction of erlotinib in non-selected non-small cell lung cancer.The retrospective study included seventy non-small-cell lung cancer patients, who were treated with erlotinib from July 2005 to July 2009. Erlotinib was prescribed at a dose of 150 mg daily.Sixty-eight patients were evaluated response. Among these patients, CR 0 case, PR 26 cases, RR (CR+PR) 38.2% and SD 24 cases as their best response, disease control rate (DCR=CR+PR+SD) 73.5%, PD 18 cases (26.5%). Sixty-three patients were evaluated PFS. The median PFS was 3.0 months. The median PFS of adenocarcinoma and non-adenocarcinoma was 3.0 months vs 2.6 months. The drug-related adverse reactions were skin rash, diarrhea and interstitial lung disease (ILD)(4.3%).Erlotinib is active in non-small cell lung cancer, and it is much more effective in adenocarcinoma and non-smoking patients. There is no difference in response or suvival concerning the sexuality. It is well tolerated in most patients.

We propose a quantitative model for dwell-based eye pointing tasks. Using the concepts of information theory to analogize eye pointing, we define an index of difficulty (IDeye) for the corresponding tasks in a similar manner to the definition that Fitts made for hand pointing. According to our validations in different situations, IDeye, which takes account of the distinct characteristics of rapid saccades and involuntary eye jitters, can accurately and meaningfully describe eye pointing tasks. To the best of our knowledge, this work is the first successful attempt to model eye gaze interactions.

In order to improve the stability of eye cursor, we introduce three methods, force field (FF), speed reduction (SR), and warping to target center (TC) to modulate eye cursor trajectories by counteracting eye jitter, which is the main cause of destabilizing the eye cursor. We evaluate these methods using two controlled experiments. One is an attention task experiment, which indicates that both FF and SR significantly alleviate the instability of eye cursor, but TC is not as we anticipated. The other is a 2D pointing task experiment, which shows that FF and SR as well as the improved implementation of SR (iSR) indeed improve human performance in dominant dwell-based eye pointing tasks of eye-based interactions. The method iSR is especially effective to accelerate eye pointing (10.5% and 8.5%) and reduce error rate (6.1% and 2.7%) when target diameter D = 45 and 60 pixels.

The idea of target prediction for pointing tasks seems to hold promise for human computer interaction. However, few studies have studied the impacts of target predictor's inner parameter settings on the human performance. In this paper, therefore, we perform an experiment to investigate the effectiveness of target prediction for pointing tasks under different parameter setting conditions. We found that parameter settings for the predictor can be optimized to improve performance. The optimal combination was sampling rate (Hz) at the level of 40 and sample size (N) at the level of 12. Our study could provide HCI researchers with some valuable insights into the subject of target prediction for pointing tasks.

In our studies, micellar electrokinetic capillary chromatography (MEKC) was employed in fingerprint analysis of Cnidium monnieri for the first time. Average chromatography of 10 batches Cnidium monnieri from Jiangsu province, China, which have long been considered as the original and genuine herbal medicine, was first established as the characteristic fingerprint. Within 25 min the major effective components were separated by 18 mM borate, 12 mM phosphate and 50 mM SDS (pH 9.2) containing 20% methanol. The relative standard deviations of migration times and peak areas were less than 5%. As a new approach of fingerprint, MKCE was compared to the conventional approach-HPLC in our experiments. The fingerprint developed by HPLC comprised 8 peaks that were collected within 40 min. Relative standard deviation (RSD) values of retention times of corresponding peaks in HPLC analysis were very small (maximum 3% and average 0.9%). In conclusion, each two methods had its advantages and disadvantages. Furthermore, besides HPLC, MEKC as a feasible method, could be used in the development of fingerprint of Cnidium monnieri.