[The emulation of clinical trials with real-world data: development and application of target trial].

Clinical trial is the gold standard for evaluating the efficacy and safety of interventions; however, it is limited by high costs and long time. Real-world data (RWD) can provide a robust data basis for comparative research, but the quality is uneven. This review introduces the target trial emulation, in which researchers, using RWD and following the design of clinical trials, define exposure and outcome in advance, set eligibility criteria, determine the time zero, estimate sample size, and plan statistical analysis, to enhance the quality of evidence for observational studies. This review preliminarily discusses the standard of evidence quality evaluation in target trial emulation. Then, the target trial emulation is shown through case interpretation.

High-energy molecular-frame photoelectron angular distributions: a molecular bond-length ruler.

We present a study on molecular-frame photoelectron angular distributions (MFPADs) of small molecules using circularly polarized synchrotron light. We find that the main forward-scattering peaks of the MFPADs are slightly tilted with respect to the molecular axis. This tilt angle is directly connected to the molecular bond length by a simple, universal formula. We apply the derived formula to several examples of MFPADs of C 1s and O 1s photoelectrons of CO, which have been measured experimentally or obtained by means of ab initio modeling. In addition, we discuss the influence of the back-scattering contribution that is superimposed over the analyzed forward-scattering peak in the case of homo-nuclear diatomic molecules such as N2.

A 2-Stage Root Analog Implant with Compact Structure, Uniform Roughness, and High Accuracy.

Immediate implant placement has the advantages of shortening the operation time, reducing the treatment cycle and cost. At present, this technology has been used widely, but the indications of immediate implantation are still limited. Here, a novel type of root analog implant (RAI) was manufactured by selective laser melting technology to address the limitation. Under optimized condition, RAIs were printed with the internal density of 99.73% and the uniform surface roughness of 11 µm (Sa). Besides, the deviation between RAI specimen and design models is controlled within 0.15 mm after optimizing scanning parameters. The substrate printed could promote human bone marrow stromal cell proliferation, spreading, and osteogenic differentiation. The bone-implant contact (BIC, 75% ± 7%) and bone volume/total volume (BV/TV, 74% ± 7%) of RAIs were significantly higher than that of conventional implants (BIC, 66% ± 5%; BV/TV, 62% ± 5%) in in vivo experiments. Further, customized abutments were designed for the RAIs, improving the masticatory ability of the beagle dogs after crown restoration. This study aims to design a personalized 2-stage RAI with compact structure and uniform roughness, in order to achieve better fracture resistance, initial osseointegration efficiency, and dispersed stress in immediate implantation. It provides a certain guiding value for standardizing the manufacture and clinical application of RAI in immediate implantation.

Diffusion MRI correlation with p16 status and prediction for tumor progression in locally advanced head and neck cancer.

Purpose: To investigate p16 effects on diffusion image metrics and associations with tumor progression in patients with locally advanced head and neck cancers. Methods: Diffusion images pretreatment and after 20 Gy (2wk) of RT were analyzed in patients with cT4/N3 p16+ oropharynx cancer (OPSCC) (N=51) and locoregionally advanced head and neck squamous cell carcinoma (LAHNSCC) (N=28), enrolled onto a prospective adaptive RT trial. Mean ADC values, subvolumes with ADC <1.2 um2/ms (TVLADC), and peak values of low (µL) and high (µH) components of ADC histograms in primary and total nodal gross tumor volumes were analyzed for prediction of freedom from local, distant, or any progression (FFLP, FFDP or FFLRDP) using multivariate Cox proportional-hazards model with clinical factors. P value with false discovery control <0.05 was considered as significant. Results: With a mean follow up of 36 months, 18 of LAHNSCC patients and 16 of p16+ OPSCC patients had progression. After adjusting for p16, small µL and ADC values, and large TVLADC of primary tumors pre-RT were significantly associated with superior FFLRDP, FFLP and FFDP in the LAHNSCC (p<0.05), but no diffusion metrics were significant in p16+ oropharynx cancers. Post ad hoc analysis of the p16+ OPSCC only showed that large TVLADC of the total nodal burden pre-RT was significantly associated with inferior FFDP (p=0.05). Conclusion: ADC metrics were associated with different progression patterns in the LAHNSCC and p16+ OPSCC, possibly explained by differences in cancer biology and morphology. A deep understanding of ADC metrics is warranted to establish imaging biomarkers for adaptive RT in HNSCC.

[Numb activates the mTORC1 signaling pathway in proximal tubular epithelial cells by upregulating V1G1 expression].

OBJECTIVE: To investigate the role of Numb in regulating mammalian target of rapamycin (mTOR) complex 1 (mTORC1) signaling pathway. METHODS: Male BALB/C mouse models of acute kidney injury (AKI) were subjected to intravenous injections of Numb-siRNA or NC-siRNA with or without intraperitoneal cisplatin injections. After the treatments, the expressions and distribution of Numb and megalin in the renal tissues of the mice were detected with immunohistochemistry, and the renal expressions of Numb, S6, p-S6, S6K1, p-S6K1, 4EBP1 and p-4EBP1 were examined with Western blotting. The proximal renal tubular epithelial cells were isolated from the mice transfected with Numb-siRNA for in vitro culture. In NRK-52E cells, the effects of amino acid stimulation, Numb knockdown, and V1G1 overexpression, alone or in combination, on expressions of Numb, S6 and p-S6 were detected with Western blotting; the expressions of AMPK and p-AMPK were also detected in transfected NRK-52E cells, mouse kidneys and cultured mouse renal tubular epithelial cells. RESULTS: In BALB/C mice, injection of Numb-siRNA caused significant reductions of Numb and p-S6 expressions without affecting megalin expression in the renal proximal tubules (P < 0.05). Cisplatin treatment obviously upregulated p-S6K1 and p-4EBP1 expressions in the kidneys of the mice (P < 0.05), and this effect was significantly inhibited by treatment with Numb-siRNA (P < 0.05). In NRK-52E cells, amino acid stimulation significantly upregulated the expression of p-S6 (P < 0.05), which was strongly suppressed by transfection with Numb-siRNA (P < 0.05). Numb knockdown inhibited AMPK activation in NRK-52E cells, mouse kidneys and primary proximal tubular epithelial cells (P < 0.05). Numb knockdown significantly downregulated V1G1 expression in NRK-52E cells (P < 0.05), and V1G1 overexpression obviously reversed the inhibitory effect of Numb-siRNA on S6 phosphorylation (P < 0.05). CONCLUSION: Numb promotes the activation of mTORC1 signaling in proximal tubular epithelial cells by upregulating V1G1 expression.

[Tumor-induced osteomalacia caused by an orbital tumor: a case report].

A 52-year-old patient was initially diagnosed as hypophosphatemic osteomalacia in the Department of Endocrinology due to knee, foot and lumbosacral pain. The symptoms were not significantly relieved after phosphorus and vitamin D supplementation. Later, the imaging examination showed an orbital tumor in the right eye. The tumor was surgically removed, and the symptoms of systemic bone pain were relieved.

[A comparative study of multiple parallel mediation analysis methods].

Objective: To introduce and compare four analysis methods of multiple parallel mediation model, including pure regression method, method based on inverse probability weighting, extended natural effect model method and weight-based imputation strategies. Methods: For the multiple parallel mediation model, the simulation experiments of three scenarios were carried out to compare the performance of different methods in estimating direct and indirect effects in different situations. Dataset from UK Biobank was then analyzed by using the four methods. Results: The estimation biases of the regression method and the inverse probability weighting method were relatively small, followed by the extended natural effect model method, and the estimation results of the weight-based imputation strategies were quite different from the other three methods. Conclusions: Different multiple parallel mediation analysis methods have different application situations and their own advantages and disadvantages. The regression method is more suitable for continuous mediator, and the inverse probability weighting method is more suitable for binary mediator. The extended natural effect model method has better performances when the residuals of two parallel mediators are positively correlated and the correlation degree is small. The weight-based imputation strategies might not be appropriate for parallel mediation analysis. Therefore, appropriate methods should be selected according to the specific situation in practice.

Saturation Transfer Difference NMR Spectroscopy for the Elucidation of Supramolecular Albumin-Polymer Interactions.

Albumin has consistently demonstrated its potential for enhancing the delivery of drugs and polymer-drug conjugates, binding via supramolecular forces within its multiple binding sites. Herein, we introduce saturation transfer difference (STD-NMR) as a method to identify the interactions between a polymer library and bovine serum albumin (BSA). With STD-NMR, the binding ability of polymers can be quickly screened by focusing on their individual structural features, making this technique more suitable for high throughput screening in comparison to traditional fluorescence studies.

Long non­coding RNA XIST promotes cell proliferation of pancreatic cancer through miR­137 and Notch1 pathway.

OBJECTIVE: Long non-coding ribonucleic acids X-inactive specific transcript (lncRNA XIST) is one lncRNAs which involved in multiple human cancers. However, the functions and potential molecular regulatory mechanisms of XIST/microRNA-137 (miR­137) in pancreatic cancer (PC) still need to explore. PATIENTS AND METHODS: PC tissues and cell lines were analyzed for XIST, miR-137 and Notch1 expressions through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Nude mouse xenograft tumor assay was used to detect XIST effects on pancreatic tumorigenesis in vivo. Cell Counting Kit (CCK-8) assay was performed to detect PC cell proliferation. Dual-Luciferase reporter assay, qRT-PCR, RNA immunoprecipitation (RIP) and Western blot assays were applied to validate the target relationship of XIST, miR­137 and Notch1. RESULTS: Results demonstrated that XIST expression was increased in PC tissues and cells. XIST knockdown inhibited PC cell proliferation in vitro and also repressed the tumor growth in vivo. XIST directly interacted with miR-137 and negatively regulated its expression. Notch1 was identified as a target gene of miR-137 and XIST acted as a competitive endogenous RNA (ceRNA) to positively regulate Notch1 expression by suppressing miR-137. In addition, we detected miR-137 was negatively correlated with XIST and Notch1 respectively, and a positive correlation between Notch1 and XIST expression in PC tissues. Furthermore, Notch1 overexpression could offset the suppressing effect of XIST knockdown or miR-137 overexpression on cell proliferation. Therefore, XIST may play an important role in promoting cell proliferation through miR­137 and Notch1 pathway in PC. CONCLUSIONS: To sum up, these results proposed that XIST functioned as an endogenous sponge in promoting PC cell proliferation through competing for miR-137 to regulate Notch1 expression, and may provide more therapeutic targets for the patients with PC.

LncRNA DSCAM-AS1 promotes non-small cell lung cancer progression via regulating miR-577/HMGB1 axis.

Non-small cell lung cancer (NSCLC) is a major source of cancer mortality. Long non-coding RNA DSCAM-AS1 has been certified to be involved in the pathogenesis of NSCLC. This study aimed to further investigate the potential mechanism of DSCAM-AS1 in NSCLC progression. The expressions of DSCAM-AS1, miR-577, and high mobility group box 1 (HMGB1) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot assay. Cell viability was assessed by Cell Counting Kit-8 (CCK-8) assay. Flow cytometry assay was conducted to monitor cell apoptosis. Cell migration and invasion were measured by transwell assay. Wnt/ß-catenin pathway-related factors were detected by western blot assay. The relationship between DSCAM-AS1, miR-577, and HMGB1 was validated by bioinformatics analysis and dual-luciferase reporter assay. The xenograft mouse model was established to analyze tumor growth in vivo. DSCAM-AS1 and HMGB1 were upregulated, while miR-577 was downregulated in NSCLC tissues and cells. DSCAM-AS1 promoted cell proliferation, migration and invasion, and restrained cell apoptosis in NSCLC cells. Overexpression of HMGB1 reversed the effects of DSCAM-AS1 depletion on the progression of NSCLC. DSCAM-AS1 modulated HMGB1 expression by sponging miR-577. DSCAM-AS1 regulated the Wnt/ß-catenin pathway by regulating miR-577 and HMGB1. DSCAM-AS1 knockdown blocked the tumor growth in vivo. In conclusion, DSCAM-AS1 facilitated NSCLC progression by regulating the HMGB1-mediated Wnt/ß-catenin pathway, providing a promising therapeutic target for NSCLC.

[Sensitivity analysis method for unmeasured confounding interference in observational study].

Objective: To introduce the methods for sensitivity analysis, discuss and compare the advantages and disadvantages of different methods. Methods: The difference between confounding function method and bounding factor method in accuracy of identifying unmeasured confounding factors in observational studies through simulation trials and actual clinical data was compared. Results: The results of simulation trials and actual clinical data showed that when there was unmeasured confounding between exposure (X) and outcome (Y), the results of confounding function and the bounding factor analysis were similar in terms of the effect of unmeasured confounding factor to lead to the complete change of the magnitude and direction of the observed effect value. However, the confounding function method needed smaller confounding effect to fully interpret the observed effect value than the bounding factor needed. In addition, the bounding factor method needed to analyze two confounding parameters, while only one parameter was needed in the confounding function method. The confounding function method was simpler and more sensitive than the bounding factor method. Conclusion: For real-world observational data, the sensitivity analysis process is essential in analyzing the causal effects between exposure (X) and outcome (Y). In terms of the calculation process and result interpretation the sensitivity analysis method of confounding function is worth to recommend.

[Estimation on the individual treatment effect among heterogeneous population, using the Causal Forests method].

Objective: This project aimed to explore the effectiveness of estimating individual treatment effect on real data, among the heterogeneous population, with Causal Forests (CF) method, to find out the characteristics of heterogeneous population. Methods: We designed and conducted four computer simulation schemes to verify the effect of estimating on individual treatment, using the CF under four different environments of the treatment effects. Real data was then analyzed for the catheterization on right heart. Results: Results from the simulation process showed that the values on individual treatment effect that were estimated by causal forests were consistent with the population effect as well as in line with the expected distribution under the setting of four different effect values. Results of real data analysis showed that values of individual treatment effect among most patients appeared positive, so the use of RHC could cause an increase of the '180-day mortality rate' in the sampled population. Patients with lower predicted probability of 2-mo survival and albumin were more likely to have a lower risk of death after using the RHC. Conclusion: CF method could be effectively used to estimate the individual treatment effect and helping the individuals to make decision on the receipt of treatment.

Knockdown of long non-coding RNA LINC00518 inhibits cervical cancer proliferation and metastasis by modulating JAK/STAT3 signaling.

OBJECTIVE: Long intergenic non-protein coding RNA 518 (LINC00518) was reported to be implicated and aberrantly expressed in multiple cancers. However, the pathogenic implications of LINC00518 in cervical cancer (CC) are still unclear. In this study, we focused on LINC00518 and investigated its expression pattern, clinical significance, and biological function in CC. PATIENTS AND METHODS: The expression levels of LINC00518 in CC tissues and cell lines were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR), and its clinical significance was assessed by statistical analysis. Cell apoptosis was determined by flow cytometry. The cell proliferation was evaluated by MTT assay and colony forming assay, and the migration and invasion were evaluated by wound healing assays and transwell assay. Western blot was used to detect the expression of relative proteins, including EMT markers and the JAK/STAT3 signaling markers. RESULTS: We found that LINC00518 was upregulated in CC tissues and associated with International Federation of Gynaecology and Obstetrics (FIGO) stage, lymph node metastasis, depth of cervical invasion and poor survival of CC patients. Univariate and multivariate Cox regression analysis showed that LINC00518 played a significant role of independent prognostic markers in overall survival rates. Furthermore, knocking down LINC00518 expression significantly suppressed CC cell proliferation, migration and invasion, and induced apoptosis in vitro. Mechanistically, the downregulation of LINC00518 suppressed JAK/STAT3 activation and subsequently decreased N-Cadherin and Vimentin. CONCLUSIONS: The present work first suggests that LINC00518 acts as an oncogene in CC via regulation of the JAK/STAT3 signaling pathway. In the future, LINC00518 may serve as a predictive biomarker and potential therapeutic target for CC patients.

Ultraviolet lasing in Zn-rich ZnO microspheres fabricated by laser ablation.

Zinc (Zn) surface plasmons (SPs) have been widely reported for their impressive performance in improving the optical properties of semiconductors. Zn is an effective metal with SPs response in the ultraviolet region, but the disadvantage of strong metal activity limits the application mentioned above. Here, in order to ensure the stability of metal Zn, ZnO/Zn microspheres were synthesized by an one-step laser ablation method to distribute Zn nanoparticles simultaneously on both inner and outer surfaces of ZnO microspheres. Lasing performance enhancement and a lower threshold were obtained in the composite which originates from the coupling between Zn SPs and the excitation light source. Accompanied by the lasing emission measurements, the coupling mechanism was explained through time-resolved photoluminescence spectroscopy (TRPL) for the samples by rapid annealing in situ. This work displays the results of lasing enhancement and the physical process of Zn SPs resonance in the ZnO/Zn microsphere.

Extra green light induced ZnO ultraviolet lasing enhancement assisted by Au surface plasmons.

We observed the dramatic enhancement of the intrinsic spontaneous and stimulated emission as well as the ensuing suppression of defect-related green emission in Au-decorated ZnO microrods. A series of spectral experiments and theoretical analysis demonstrated an electron transfer assisted process by surface plasmon (SP) resonant coupling between the Au nanoparticles and ZnO. The mechanism indicates an approach to enhance the UV emission of ZnO through an extra excitation of visible light similar to that for the defect emission of ZnO. Based on the coupling mechanism, the externally enhanced ultraviolet lasing was further improved from 1.5 to 2.8-fold by adjusting the pumping power of the green light intensity in the Au/ZnO hybrid cavity. This research not only further confirms the SPR-assisted electron transfer process but also offers an approach to improve the intrinsic UV emission even for heavily-defected ZnO through visible light excitation via a nonlinear process.

A direct role for hepatitis B virus X protein in inducing mitochondrial membrane permeabilization.

Hepatitis B virus X protein (HBx) acts as a multifunctional protein that regulates intracellular signalling pathways during HBV infection. It has mainly been studied in terms of its interaction with cellular proteins. Here, we show that HBx induces membrane permeabilization independently of the mitochondrial permeability transition pore complex. We generated mitochondrial outer membrane-mimic liposomes to observe the direct effects of HBx on membranes. We found that HBx induced membrane permeabilization, and the region comprising the transmembrane domain and the mitochondrial-targeting sequence was sufficient for this process. Membrane permeabilization was inhibited by nonselective channel blockers or by N-(n-nonyl)deoxynojirimycin (NN-DNJ), a viroporin inhibitor. Moreover, NN-DNJ inhibited HBx-induced mitochondrial depolarization in Huh-7 cells. Based on the results of this study, we can postulate that the HBx protein itself is sufficient to induce mitochondrial membrane permeabilization. Our finding provides important information for a strategy of HBx targeting during HBV treatment.

Long-term Outcomes for Liver Transplant Recipients in Terms of Hepatic Encephalopathy.

BACKGROUND: Liver transplantation (LT) is thought to resolve cognitive deficit due to hepatic encephalopathy (HE). The aim of this study was to determine the factors associated with the outcomes of patients with HE after LT. METHODS: The authors reviewed the medical records of 388 patients with HE who underwent LT from 1996 to 2014. RESULTS: There were 282 patients with grade 1-2 HE and 106 patients classified as grade 3-4. Patients in the latter group had a tendency for a more decompensated hepatic condition than patients with grade 1-2 HE. HE sequelae were only associated with grade 3-4 HE with borderline significance (P = .05). The cumulative 1-, 3-, and 5-year overall survival (OS) of patients with grade 1-2 HE were 81.9%, 77.3%, and 74.6%, whereas those of in patients with grade 3-4 HE were 77.4%, 73.3%, and 72.2%, respectively (P = .75). CONCLUSION: The sequelae of HE were only associated with the grade 3-4 HE. Aggressive treatment of HE prior to LT may prevent patients from deteriorating into high-grade HE, which could further contribute to improving the outcomes after LT.