Establishment and application of a flow cytometry-based method for detecting histone acetylation levels.

Histone deacetylase inhibitors, which have also received attention in AIDS and other diseases, are a new class of anticancer drugs developed in recent years. However, there is still a lack of a unified and reliable method for detecting histone acetylation levels in basic and clinical research. In this study, we developed a flow cytometry-based method to detect histone acetylation levels by comparing different sample processing temperature (on ice vs. room temperature), permeabilization method (intracellular vs. nuclear), antibody dose (antibody titration) and antibody incubation time (time gradient) using whole blood and peripheral blood mononuclear cells. In addition, we applied this optimized method in in vitro experiment and clinical trial of Chidamide (the only China FDA approved HDACi), the result of which confirmed that the flow cytometry-based method for detecting histone acetylation levels is a reliable, fast and convenient method which can be used in basic and clinical research.

A richly interactive exploratory data analysis and visualization tool using electronic medical records.

BACKGROUND: Electronic medical records (EMRs) contain vast amounts of data that is of great interest to physicians, clinical researchers, and medial policy makers. As the size, complexity, and accessibility of EMRs grow, the ability to extract meaningful information from them has become an increasingly important problem to solve. METHODS: We develop a standardized data analysis process to support cohort study with a focus on a particular disease. We use an interactive divide-and-conquer approach to classify patients into relatively uniform within each group. It is a repetitive process enabling the user to divide the data into homogeneous subsets that can be visually examined, compared, and refined. The final visualization was driven by the transformed data, and user feedback direct to the corresponding operators which completed the repetitive process. The output results are shown in a Sankey diagram-style timeline, which is a particular kind of flow diagram for showing factors' states and transitions over time. RESULTS: This paper presented a visually rich, interactive web-based application, which could enable researchers to study any cohorts over time by using EMR data. The resulting visualizations help uncover hidden information in the data, compare differences between patient groups, determine critical factors that influence a particular disease, and help direct further analyses. We introduced and demonstrated this tool by using EMRs of 14,567 Chronic Kidney Disease (CKD) patients. CONCLUSIONS: We developed a visual mining system to support exploratory data analysis of multi-dimensional categorical EMR data. By using CKD as a model of disease, it was assembled by automated correlational analysis and human-curated visual evaluation. The visualization methods such as Sankey diagram can reveal useful knowledge about the particular disease cohort and the trajectories of the disease over time.

CD147-spike protein is a novel route for SARS-CoV-2 infection to host cells.

In face of the everlasting battle toward COVID-19 and the rapid evolution of SARS-CoV-2, no specific and effective drugs for treating this disease have been reported until today. Angiotensin-converting enzyme 2 (ACE2), a receptor of SARS-CoV-2, mediates the virus infection by binding to spike protein. Although ACE2 is expressed in the lung, kidney, and intestine, its expressing levels are rather low, especially in the lung. Considering the great infectivity of COVID-19, we speculate that SARS-CoV-2 may depend on other routes to facilitate its infection. Here, we first discover an interaction between host cell receptor CD147 and SARS-CoV-2 spike protein. The loss of CD147 or blocking CD147 in Vero E6 and BEAS-2B cell lines by anti-CD147 antibody, Meplazumab, inhibits SARS-CoV-2 amplification. Expression of human CD147 allows virus entry into non-susceptible BHK-21 cells, which can be neutralized by CD147 extracellular fragment. Viral loads are detectable in the lungs of human CD147 (hCD147) mice infected with SARS-CoV-2, but not in those of virus-infected wild type mice. Interestingly, virions are observed in lymphocytes of lung tissue from a COVID-19 patient. Human T cells with a property of ACE2 natural deficiency can be infected with SARS-CoV-2 pseudovirus in a dose-dependent manner, which is specifically inhibited by Meplazumab. Furthermore, CD147 mediates virus entering host cells by endocytosis. Together, our study reveals a novel virus entry route, CD147-spike protein, which provides an important target for developing specific and effective drug against COVID-19.

A case of chylothorax treated curatively with Sapylin, a streptococcus preparation.

Chylothorax is an uncommon disease where fatty fluid accumulates within the chest cavity. Conservative management, including repeated thoracentesis or pleurodesis, seems to be suitable to most cases. Herein, we present a case of efficacious pleurodesis by intrapleural injection of Sapylin, a streptococcus preparation, for the treatment of chylothorax. A 52-year-old non-smoking female farmer was diagnosed as idiopathic chylothorax after we ruled out possible causes including chest trauma, lymphoma, lung cancer, filariasis, tuberculosis, and etc. Two-time intra-thoracic injection of 3 Klinische Einheit (KE) Sapylin achieved rapid and effective control of chylothorax with no severe side effects. Sapylin may facilitate pleurodesis by producing a strong inflammatory response.

A novel tool for visualizing chronic kidney disease associated polymorbidity: a 13-year cohort study in Taiwan.

OBJECTIVE: The aim of this study is to analyze and visualize the polymorbidity associated with chronic kidney disease (CKD). The study shows diseases associated with CKD before and after CKD diagnosis in a time-evolutionary type visualization. MATERIALS AND METHODS: Our sample data came from a population of one million individuals randomly selected from the Taiwan National Health Insurance Database, 1998 to 2011. From this group, those patients diagnosed with CKD were included in the analysis. We selected 11 of the most common diseases associated with CKD before its diagnosis and followed them until their death or up to 2011. We used a Sankey-style diagram, which quantifies and visualizes the transition between pre- and post-CKD states with various lines and widths. The line represents groups and the width of a line represents the number of patients transferred from one state to another. RESULTS: The patients were grouped according to their states: that is, diagnoses, hemodialysis/transplantation procedures, and events such as death. A Sankey diagram with basic zooming and planning functions was developed that temporally and qualitatively depicts they had amid change of comorbidities occurred in pre- and post-CKD states. DISCUSSION: This represents a novel visualization approach for temporal patterns of polymorbidities associated with any complex disease and its outcomes. The Sankey diagram is a promising method for visualizing complex diseases and exploring the effect of comorbidities on outcomes in a time-evolution style. CONCLUSIONS: This type of visualization may help clinicians foresee possible outcomes of complex diseases by considering comorbidities that the patients have developed.

Single photon emission computed tomography (SPECT) for low back pain induced by extension with no root sign.

BACKGROUND: Low back pain (LBP) is very common in the general population. Most patients with LBP will receive an X-ray examination on lumbar spine; however, the results are likely to show a negative finding or degenerative joint disease, which are not truly pathological factors. Among various imaging diagnostic tools for active bony lesions of lumbar spine, planar bone scintigraphy has a higher sensitivity, but its ability to locate anatomic lesions is less satisfactory. The purpose of this study was to investigate the role of SPECT for evaluation of LBP. METHODS: Fifty-two consecutive patients who had low back pain induced by extension were studied. They had no evidence of malignant tumor, immune disease, spinal infection and neurological disorder by history-taking and physical examination. All patients received planar bone scintigraphy and SPECT exams following an X-ray examination. The results of X-ray finding were grouped into 3 categories: (A) normal; (B) degenerative joint arthritis; (C) spondylolysis. The data of test results and clinical evaluation were then used for analysis. RESULTS: Twenty (38.5%) out of 52 patients examined by planar scan had abnormality, with 29 increased uptake lesions, compared with 28 (53.8%) out of 52 patients with 60 increased uptake lesions by SPECT with planar scan; SPECT disclosed 1-2 more lesions with improved location in 15 patients (p < 0.05). Of the 52 patients, 21 (40.4%) presented in group A, 21 (40.4%) in group B, and 10 (19.2%) in group C according to the X-ray examination. In group A, 9 out of 21 (42.9%) patients had an abnormal SPECT result, compared to 5 of 10 (50%) in group B, and 14 out of 21 (66.7%) in group C, respectively (p > 0.05). The location of abnormal uptake on L-spine included vertebral body and arch (57.1%), vertebral arch (28.6%), and vertebral body (14.3%). Most of lesions (91.5%) were distributed at the 4th and 5th lumbar vertebral segments. CONCLUSIONS: SPECT was more sensitive and located more lesions than planar bone scintigraphy, especially when the lesions were located at posterior element of vertebrae. Most of the lesions were distributed at the 4 th and 5 th lumbar vertebral segments. There was no significant statistical difference of abnormal SPECT related to X-ray finding. The use of SPECT was the first choice among all image modalities when cause of low back pain was assumed to arise from bone and joint disorder at clinical evaluation.

Cyclic alphavbeta6-targeting peptide selected from biopanning with clinical potential for head and neck squamous cell carcinoma.

BACKGROUND: A cyclic peptide-displaying phage library was used for biopanning on oral squamous cell carcinoma (OSCC) cells to identify cancer-targeting peptides. This study was designed to characterize the receptor specificity of a candidate phage clone/peptide (phage/peptide-29) and to explore the clinical potential of this peptide. METHODS: Immunofluorescent confocal microscopy, phage binding assay, and immunohistochemical studies were used to demonstrate the receptor specificity of phage/peptide-29. The effect of peptide-29 on the proliferation of OSCC cells was studied using 3-dimensional (3D) cell cultures. RESULTS: Phage/peptide-29 preferentially binds integrin alphavbeta6 rather than other alphav-associated integrins. Peptide-29 significantly inhibits the proliferation of OSCC cells in 3D cell cultures. On human pathological sections, phage-29 targets oral cancer cells in a alphavbeta6-dependent manner. Besides, we showed that integrin alphavbeta6 is universally (94.7%, 36/38) expressed in all major kinds of head and neck squamous cell carcinomas (HNSCC). CONCLUSIONS: Peptide-29 selected from biopanning may have clinical potential for HNSCC.