Causal mediation analysis investigates whether the effect of an exposure on an outcome operates through intermediate variables known as mediators. Although progress has been made in high-dimensional mediation analysis, current methods do not reliably control the false discovery rate (FDR) in finite samples, especially when mediators are moderately to highly correlated or follow non-Gaussian distributions. These challenges frequently arise in DNA methylation studies. We introduce CoxMDS, a multiple data splitting method that uses Cox proportional hazards models to identify putative causal mediators for survival outcomes. CoxMDS ensures finite-sample FDR control even in the presence of correlated or non-Gaussian mediators. Through simulations, CoxMDS is shown to maintain FDR control and achieve higher statistical power compared with existing approaches. In applications to DNA methylation data with survival outcomes, CoxMDS identified eight CpG sites in The Cancer Genome Atlas that are consistent with the hypothesis that DNA methylation may mediate the effect of smoking on lung cancer survival, and two CpG sites in the Alzheimer's Disease Neuroimaging Initiative that are consistent with the hypothesis that DNA methylation may mediate the effect of smoking on time to Alzheimer's disease conversion.

Hepatocellular carcinoma (HCC) is one of the most common cancers in the world. Alpha-fetoprotein (AFP) is one of the most important diagnostic markers for HCC. However, over 30% of HCC patients are negative for AFP (< 20 ng/mL), emphasizing the great need for new biomarkers for early diagnosis. A cohort of 100 AFP-negative early-stage HCC patients and 100 controls consisting of a discovery group and two validation groups was constructed. UPLC-MS/MS was performed on serum to identify diagnostic biomarkers. 17 differential metabolites were identified as diagnostic candidates for AFP-negative early-stage HCC. KEGG pathway enrichment analysis showed that glycine, serine, threonine and linoeic acid metabolism pathways were significantly enriched. After ranking feature weight of differential metabolites, Aspartic acid and 11Z-Eicosenoic Acid were selected to construct the prediction model. Logistic regression, random forest and support vector machine validation assessed high diagnostic performance of the prediction model (AUC=0.981). This study provides novel insights into early diagnosis of AFP-negative HCC, and is expected to improve the outcomes of ESCC.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-associated mortality worldwide. Alpha-fetoprotein (AFP) is the most commonly used diagnostic marker for HCC. However, over 30% of HCC patients are negative for AFP ( < 20 ng/mL), which makes early diagnosis difficult and leads to complex heterogeneity and poor prognosis. Therefore, AFP-negative-specific heterogeneity is of great value in HCC study. Hepatocellular carcinoma (HCC) is the third leading cause of cancer-associated mortality worldwide. Alpha-fetoprotein (AFP) is the most commonly used diagnostic marker for HCC. However, over 30% of HCC patients are negative for AFP ( < 20 ng/mL), which makes early diagnosis difficult and leads to complex heterogeneity and poor prognosis. Therefore, AFP-negative-specific heterogeneity is of great value in HCC study. Quantitative proteomic profiling was performed to characterize paired tumor and non-tumor tissues of 124 AFP-negative HCC patients and 76 AFP-positive HCC patients. The proteomic data showed the specific proteins expression profile of AFP-negative HCC. Furthermore, we identified heterogeneity in AFP-negative HCC, and classified the cohort into the subtype I, II and III, which showed significant difference in clinical outcome. Subtype II, which is characterized by immunoreactivity and proliferation, is associated with the shortest overall survival and disease-free survival. Meanwhile, based on the patient-derived tumor organoids and xenograft mouse model of HCC, we found that the above stratification is valuable on distinguishing individual differences in patients response to chemotherapeutic and molecular targeted agents. Through this study, the proteomic stratification of AFP-negative HCC yielded new insight into the tumor biology, and provided new possibility for personalized therapies.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death. Early HCC is mainly resected by surgery, but some patients have recurrence and metastasis within 5 years after surgery. Therefore, it is essential to explore the mechanism of liver cancer metastasis and find potential intervention methods for liver cancer. DNA methylation is a form of chemical modification of DNA that can alter genetic performance without changing the DNA sequence. A large number of studies have shown that DNA methylation is an important cause of the occurrence and development of liver cancer. SERPINE2, known as PN-1, is a secreted protein with anti-serine protease activity against serine proteases such as thrombin, urokinase, and plasminogen. Previous studies have shown that alterations in SERPINE2 expression contribute to tumorigenesis and participate in tumor metastasis. The differential expression of SERPINE2 between cancer tissues and normal tissues was analyzed using TCGA database. The correlation between SERPINE2 and clinical data was also analyzed by TCGA database. SERPINE2-knockdown and SERPINE2-overexpression HCC cell lines were constructed using lentiviruses and using transwell assay and wound healing assay to investigate the effect of SERPINE2 on HCC metastasis. Mouse model of lung colonization was established by tail vein injection of lentivirus-stabilized SERPINE2-overexpressing HCC cells. Firstly, we found that DNA methylation of SERPINE2 is regulated by DNMT1, and the methylation level of SERPINE2 was significantly inhibited and the expression of SERPINE2 was up-regulated after AZA treatment in HCC cell lines. SERPINE2 is highly expressed in HCC and closely associated with prognosis and other clinical features of HCC, patients with high SERPINE2 expression tend to have poor prognosis, and we found that the expression level of SERPINE2 was closely related to tumor stage, the worse the tumor stage and the higher the degree of malignancy, the higher the expression level of SERPINE2. Notably, in patients with microvascular metastasis, those with high SERPINE2 expression tended to have a worse prognosis. Overexpression of SERPINE2 promotes HCC cell metastasis in vitro and in vivo. Furthermore, SERPINE2 also affects tumor cell adhesion and extracellular matrix remodeling. In conclusion, our study has demonstrated that SERPINE2 expression is regulated by DNA methylation, and its expression is significantly higher in HCC tissues than in normal tissues, and alterations of SERPINE2 expression can significantly affect tumor cell migration, invasion, and lung colonization. In addition, we also demonstrated that SERPINE2 expression can affect cell adhesion ability and extracellular matrix remodeling. SERPINE2 is expected to be a new target for the treatment of HCC metastasis.

This study highlights the use of a newly developed ESAPI script specific to brachytherapy plans which allows for iteration over multiple patients at one time. We were able to quickly compare multiple dose metrics for two patient groups treated at our institution.

Zellweger syndrome (ZS) is the most severe phenotype in peroxisomal biogenesis disorders (PBDs). 14 Peroxin (PEX) genes have been identified to attribute PBDs, and about 70% of the ZS patients harbor gene mutations in PEX1. Recently gene mutation screening combined with clinical manifestations such as distinct facial features and congenital malformations is considered a suitable test for ZS patients. Here, a Chinese newborn patient with clinical features of ZS confirmed by molecular findings was reported. A novel pathogenic variation of the PEX1 gene was identified by exome sequencing. The patient is a homozygote of c.1671_1672delAG variation in the PEX1 gene and was inherited from her heterogenous parents, respectively. This variation leads to early termination of translation and produces a non-functional truncated protein. We report a novel pathogenic variation in the PEX1 gene, providing valuable information for genetic counseling and reproductive options.

The origin of cancer is related to the dysregulation of multiple signal pathways and of physiological processes. Bromodomain-containing protein 4 (BRD4) has become an attractive target for the development of anticancer and anti-inflammatory agents since it can epigenetically regulate the transcription of growth-promoting genes. The synthesized BRD4 inhibitors with new chemical structures can reduce the drug resistance, but their binding modes and the inhibitory mechanism remain unclear. Here, we initially constructed robust QSAR models based on 68 reported tetrahydropteridin analogues using topomer CoMFA and HQSAR. On the basis of QSAR results, we designed 16 novel tetrahydropteridin analogues with modified structures and carried out docking studies. Instead of significant hydrogen bondings with amino acid residue Asn140 as reported in previous research, the molecular docking modelling suggested a novel docking pose that involves the amino acid residues (Trp81, Pro82, Val87, Leu92, Leu94, Cys136, Asp144, and Ile146) at the active site of BRD4. The MD simulations, free energy calculations, and residual energy contributions all indicate that hydrophobic interactions are decisive factors affecting bindings between inhibitors and BRD4. The current study provides new insights that can aid the discovery of BRD4 inhibitors with enhanced anti-cancer ability.

This cohort study assesses the utility of restricted mean survival time as a method for quantifying time to nursing home placement among patients with dementia.

Based on the software Visual-Environment, Finite element method (FEM) was performed on the dissimilar butt-joint between Q345 and 2Cr13 steel aiming at the welding residual stress. The main contents in the paper were different current intensity was applied to modeling the welding process of Q345/2Cr13 dissimilar steel and the law of residual stress field were discussed. Based on the result, different current intensities have little effect on the lateral residual stress, while the longitudinal residual stress and the initial and end of the weld have a great influence. The physical properties of the dissimilar plates lead to uneven distribution of residual stress, and the current intensity should be smaller.

Investigation and analysis of CAR-T’s market access and reimbursement strategy in China. Literature review of concerned publications from July 2018 to December 2019 was conducted and 38 articles were included. Further in-depth interview for different stakeholders was performed, including health insurance experts, academic researchers, physicians and pharmacists, with a total of 6 visitors. And two advisory board meetings were organized among relevant experts from the medical insurance department, the health department and the hospital management department, with a total of 26 experts participating the seminar. All the materials were compiled for analysis. 1. Commercial health insurance could be applied firstly to build up the CAR-T’s market access and reimbursement strategy, and gradually to touch the public primary medical insurance system in China. A multi-level market access strategy could be set here. 2. Evidence-based health technology assessment (HTA)plays an important role in market access for expensive healthcare program like CAR-T. Enterprises should pay attention to this and provide high-quality evidence and dossier in market access. 3. Due to complicated administration needed for performance-based payment, it could be very hard to implement in China. Instead, budget risk sharing program would be much more feasible in China by negotiating with National Healthcare Security Administration on appropriate price, strict indication, certified hospital and annual budget ceiling, etc. Innovative and feasible strategy is essential for CAR-T to get market access in China.

Based on the numerical simulation software Visual-Environment, the numerical calculation and analysis of residual stress field under different preheating temperatures for Q345/2Cr13 dissimilar plate welding were carried out in this paper. The effects of different preheating temperatures on post-weld residual stress were mainly studied. The results showed that different preheating temperatures have little effect on the lateral residual stress, while the longitudinal residual stress and the initial and end of the weld have greater impacts. For residual stress difference between the two base metals should not be too high, the dissimilar plate welding should adopt a moderate preheating temperature.

Adalimumab (ADM) serum levels (SL) during maintenance are associated with treatment outcome and need for dose-escalation in Crohn’s disease (CD) patients. Little is known about the clinical relevance of proactive testing of ADM SL during induction. We evaluated correlation between ADM SL at Week 4, ADM anti-drug antibody (ADA) presence and outcome. Serum samples from biologically naïve CD patients were prospectively collected at trough at Weeks 4 and 12 after ADM initiation. Clinical remission was defined as an average daily stool frequency ≤2.8 and an average abdominal pain score ≤1. In patients with an elevated baseline C-reactive protein (CRP), biochemical remission was defined as a CRP ≤5.0 mg/l and response as a decrease of at least 50% or CRP normalisation. ADM SL were measured with a novel ADM RIDA®QUICK lateral flow assay (LFA, R-biopharm) and benchmarked with the RIDASCREEN® ELISA. ADA presence was determined using a drug-resistant assay, allowing detection in presence of high concentrations of ADM.1 Ninety-two patients with active CD (median disease duration 3.3 years) were included. Median SL at Week 4, measured by LFA (11.2 µg/ml, IQR 8.2–14.5), correlated well with median ELISA SL at Week 4 (10.0 µg/ml, IQR 7.7–12.9) (r = 0.96, p < 0.001). Lower median SL at Week 4 were significantly associated with the presence of ADA at Week 12 (8.4 in ADA positive vs. 12.8 µg/ml in ADA negative patients, p = 0.006). Only 3 out of 15 ADA+ patients at Week 12, had detectable ADA at Week 4 already. Similarly, a trend towards lower median SL at Week 2 could be observed in these 3 ADA+ patients at Week 4, compared with the remaining ADA patients (4.4 vs. 9.9 µg/ml, p = 0.2). Although median weighted patient reported outcome, PRO2, significantly decreased from baseline to Week 12 (15.0 vs. 8.0, p < 0.001), SL at Week 4 were not significantly associated with clinical remission at Week 12. However, SL at Week 4 were associated with biological response and remission at Week 12 (p = 0.002, p = 0.005), and with the need for dose-escalation in symptomatic patients within the first year (p = 0.01). In patients given dose-escalation, discontinuation of ADM thereafter, due to loss-of-response (LOR), was associated with lower SL at Week 12 (p = 0.02) and ADA positivity at Week 12 (p = 0.001). Lower SL at Week 4 are associated with ADA development later on, and seem to be the cause rather than the consequence of lower SL afterwards. ADM SL during induction may predict the need for and the success of dose-escalation. Although these findings need prospective validation, availability of an ADM rapid assay creates the opportunity for optimising therapy early during induction. 1. Bian S, Ferrante M, Gils A. Validation of a drug-resistant anti-adalimumab antibody assay to monitor immunogenicity in the presence of high concentrations of adalimumab. AAPS, 2017;468–474.

An association between vedolizumab (VDZ) trough concentrations and outcome has been observed in patients with inflammatory bowel diseases. This association was more pronounced in patients with ulcerative colitis (UC) compared with Crohn’s disease (CD). We aimed to develop and validate a dried blood spot (DBS) sampling method to facilitate intensive sampling for exploring the pharmacokinetics of VDZ in more detail. First, DBS were prepared through spotting of 40 µL of whole citrated blood spiked with VDZ (2–50 µg/ml) onto a Protein Saver Card. Blood was extracted from DBS cards and the extracts were analysed on ELISA. In addition to routine method validation (precision, accuracy, sensitivity, selectivity), DBS-related parameters including blood volumes, storage stability and impact of haematocrit were also assessed. Second, DBS derived from finger prick and serum samples obtained via venipuncture were taken concurrently at trough from 15 patients (7 UC and 8 CD) on at least one occasion and VDZ concentrations were compared. Statistical analyses were performed using R. Spiking VDZ to citrated whole blood followed by DBS sampling and extraction revealed an average extraction efficiency of 70 ± 2% (n = 23) with an accuracy of 98–104% and an imprecision of 7–11% for each concentration analysed. Residual anti-TNF and antibodies towards anti-TNF did not impact VDZ concentration whereas the addition of anti-VDZ antibodies to spiked VDZ samples caused a similar decrease in VDZ concentration in the DBS-based as in the serum-based measurements. Blood spot volumes between 15 µL and 50 µL produced comparable results. Storing the DBS papers at room temperature for one month or the extracts at -20°C for 3 months did not impair DBS recovery (within 80–120% compared with the first measurements). Median VDZ serum-to-DBS ratio of 2.03 (IQR 1.89–2.01; Spearman’s rank rho=0.93, p < 0.0001) was obtained across the therapeutic relevant range (7.5–39 µg/ml serum concentration, 17-paired patient samples). DBS-converted serum concentrations showed no significant differences with analysed serum concentrations (p = 1.00; Figure1). No analytically relevant impact of haematocrit was observed in the range of 33.6% to 48.9%. The correlation (left panel) and differences (right panel) between DBS-converted vedolizumab serum concentrations and analysed vedolizumab serum concentrations. VDZ blood concentrations highly correlate with VDZ serum concentrations over a broad concentration range. The developed tool and the derived conversion ratio can be used to perform VDZ monitoring with improved flexibility by sampling at home, patient convenience and robustness.

Objective We aimed to investigate the clinical features of acute acalculous cholecystitis (AAC) in patients with systemic lupus erythematosus (SLE). Methods SLE patients with AAC hospitalized in the Peking Union Medical College Hospital (PUMCH) from January 2001 to September 2015 were retrospectively analyzed. Their medical records were systematically reviewed. The diagnosis of AAC was based on clinical manifestations and confirmed by radiologic findings including a distended gallbladder with thickened wall, pericholecystic fluid and absence of gallstones. Results Among the 8411 hospitalized SLE patients in PUMCH, 13 (0.15%) were identified to have SLE-AAC. Eleven (84.6%) of them were female, with a mean age of 30.1 ± 8.6 years. AAC was the initial manifestation of SLE in four (30.8%) cases. Eleven (84.6%) patients complained of fever and abdominal pain, four (30.8%) had positive Murphy’s sign and six (46.2%) had elevated liver enzymes. The median SLE Disease Activity Index was 8.0 (range 0–20.0) at the time of AAC. Other affected organs in SLE-AAC included kidney (11, 84.6%) and hematologic system (11, 84.6%), followed by mucocutaneous (seven, 53.8%), musculoskeletal (seven, 53.8%) and neuropsychiatric (two, 15.4%) systems. All patients received treatment of glucocorticoids and immunosuppressants but none underwent surgical intervention. During a median follow-up of 28 months (range, 2–320 months), 12 cases (92.4%) responded to treatment with no relapse and one patient (7.6%) died of septic shock. Conclusion Our study suggests that AAC is a relatively uncommon and underestimated gastrointestinal involvement of SLE that is often associated with active disease. For patients with AAC in SLE, treatment with aggressive glucocorticoids could result in a good prognosis.

The existence of clearance in joints of mechanism is inevitable. In this paper, the friction effects in clearance joints on dynamic responses of driving mechanism of satellite antenna are studied. Considering clearances in joints, the contact force model in clearance joints is established using a nonlinear continuous contact force model and the friction effect is considered by using a modified Coulomb friction model. Then the dual-axis driving mechanism of satellite antenna with clearance joints is used as the application example. The numerical simulation of dual-axis driving mechanism with clearance joints is presented. The friction effects of clearance joint on dynamic responses of the dual-axis driving mechanism are discussed and analyzed quantitatively for four cases with different friction coefficients. The investigation results show that the increase of friction coefficient will decrease the vibration amplitude of the driving mechanism system.