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Xian-Ping Yang,1,* Ying-Yao Liu,2,* Cai-Yun Zhang,3 Kai-Kai Huang,1 Shan-Shan Han,1 Bao-Ying Liang,1 Ying Lin1 1Department of Dermatology, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine (Guangdong Provincial Hospital of Chinese Medicine), Guangzhou, Guangdong, People’s Republic of China; 2The Second Clinical Medical College, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, People’s Republic of China; 3Department of Dermatology, Dongguan Traditional Chinese Medicine Hospital, Dongguan, Guangdong, People’s Republic of China*These authors contributed equally to this workCorrespondence: Ying Lin, Department of Dermatology, the Second Affiliated Hospital of Guangzhou University of Chinese Medicine (Guangdong Provincial Hospital of Chinese Medicine), No. 111 Dade Road, Guangzhou, Guangdong, 510120, People’s Republic of China, Email lin19791226@gzucm.edu.cnAim: Atopic dermatitis (AD) often accompanies skin infections, and bacterial skin infections often cause persistent and worsening symptoms. In this study, we explored the key changes in the microbiota of AD patients, as well as the effects of different ages and the severity of rash on changes in the microbiota.Patients and Methods: A total of 95 AD patients and 77 healthy volunteers were recruited. The AD patients were divided into three groups based age and three groups according to the EASI score. Microorganisms collected from the skin were analyzed through 16S rRNA gene sequencing, revealing species diversity via α and β diversity analyses. Species compositions were compared at the phylum and genus levels. The significance of skin microbiota at the genus level was assessed using the random forest algorithm. Finally, the impact of relationships between different microbial communities on the microbial community composition and the pathogenesis of AD was explored using Pearson correlation coefficients.Results: The species diversity of the skin microbiota in the AD group significantly decreased. Compared with that in the healthy volunteers (HV) group, the bacterial diversity in the two groups of samples significantly differed. Staphylococcus dominated the bacterial communities, and as AD symptoms gradually worsened, the abundance of Staphylococcus gradually increased. Among all bacterial genera with a relative abundance greater than 1%, Staphylococcus showed a negative correlation with other genera, and showed significant consistency in specimens from different age groups.Conclusion: Changes in the abundance of Staphylococcus in the skin bacterial colonies are the main cause of AD. Brevundimonas, Paracoccus, Corynebacterium, and Veillonella may serve as characteristic biomarkers for AD. These results indicate that altering the microbiota composition of the skin may aid in the treatment of AD.Keywords: atopic dermatitis, skin bacterial colony, 16S rRNA gene sequencing

Epiphyseal region is the most sensitive region to developmental changes of the skeletal system. Extraction of this area is the very first step in any computerized image analysis. In this report a fully automated analysis of a hand radiograph resulting in extraction of distal and middle regions of the II, III, and IV phalanx is presented. The processing is performed in 3 stages. First, the trend of background is removed from radiograph to obtain a binary hand mask. At this stage a labeling procedure is necessary to eliminate artifacts (markers). Then, II, III, and IV phalanges are identified in the binary image, and the phalangeal axes are drawn. Finally, the intensity profile along each phalangeal axis is analyzed, and, on its basis, distal and middle regions are located. The presented procedure is designed as a part of currently developed system for automatic bone age assessment; however, it also can be as a preprocessing step in other diseases the diagnoses of which may require a computer assistance. Copyright © 2001 by W.B. Saunders Company [ABSTRACT FROM AUTHOR]

Introduction: Pressurized intraperitoneal aerosol chemotherapy-oxaliplatin (PIPAC-OX) induces direct DNA damage and immunogenic cell death in patients with gastric cancer peritoneal metastases (GCPM). Combining PIPAC-OX with immune checkpoint inhibition remains untested. We conducted a phase I first-in-human trial evaluating the safety and efficacy of PIPAC-OX combined with systemic nivolumab (NCT03172416)., Methods: Patients with GCPM who experienced disease progression on at least first-line systemic therapy were recruited across three centers in Singapore and Belgium. Patients received PIPAC-OX at 90 mg/m 2 every 6 weeks and i.v. nivolumab 240 mg every 2 weeks. Translational studies were carried out on GCPM samples acquired during PIPAC-OX procedures., Results: In total, 18 patients with GCPM were prospectively recruited. The PIPAC-OX and nivolumab combination was well tolerated with manageable treatment-related adverse events, although one patient suffered from grade 4 vomiting. At second and third PIPAC-OX, respectively, the median decrease in peritoneal cancer index (PCI) was -5 (interquartile range: -12 to +1) and -7 (interquartile range: -6 to -20) and peritoneal regression grade 1 or 2 was observed in 66.7% (6/9) and 100% (3/3). Translational analyses of 43 GCPM samples revealed enrichment of immune/stromal infiltration and inflammatory signatures in peritoneal tumors after PIPAC-OX and nivolumab. M2 macrophages were reduced in treated peritoneal tumor samples while memory CD4+, CD8+ central memory and naive CD8+ T-cells were increased., Conclusions: The first-in-human trial combining PIPAC-OX and nivolumab demonstrated safety and tolerability, coupled with enhanced T-cell infiltration within peritoneal tumors. This trial sets the stage for future combinations of systemic immunotherapy with locoregional intraperitoneal treatments., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The present study aims to screen and evaluate the early clinical predictors for type 2 diabetes mellitus (T2DM) patients with mild cognitive impairment (MCI) and dementia in Hunan province. A cross-sectional study was conducted from May 2023 to October 2023 to collect data on long-term T2DM patients who settled in Hunan province and were treated in the Department of Geriatrology at Xiangya Hospital of Central South University. The patients were grouped according to the Montreal Cognitive Assessment (MoCA) scale. Basic patient information and multiple serum markers were collected, and differences between groups were compared using one-way ANOVA or Kruskal-Wallis (KW) tests. The multivariate logistic regression analysis was utilized to assess risk factors and Nomogram models were constructed. The logistic regression analysis showed that years of education and serum levels of 1, 5-AG were related factors for the progression of T2DM to T2DM with MCI, and body weight, years of education and FPN levels affected the progression of T2DM with MCI to T2DM with dementia. Based on this, two Nomogram risk prediction models were established. The area under the curve (AUC) of the Nomogram model predicting T2DM progression to T2DM combined with MCI was 0.741, and the AUC of the Nomogram model predicting T2DM combined with MCI progression to T2DM combined with dementia was 0.734. The calibration curves (DCA) of the two models in the training and validation sets were symmetrically distributed near the diagonal line, indicating that the models in the training and validation sets could match each other. In summary, body weight, years of education, and serum HDL-3, FPN, and 1, 5-AG levels are associated with the development of MCI and dementia in T2DM patients. The Nomogram models constructed based on these factors can predict the risk of MCI and dementia in T2DM patients, providing a basis for clinical decision-making.

Enhancer cis -regulatory elements play critical roles in gene regulation at many stages of cell growth. Enhancers in cancer cells also regulate the transcription of oncogenes. In this study, we performed a comprehensive analysis of long-range chromatin interactions, histone modifications, chromatin accessibility and expression in two gastric cancer (GC) cell lines compared to normal gastric epithelial cells. We found that GC-specific enhancers marked by histone modifications can activate a population of genes, including some oncogenes, by interacting with their proximal promoters. In addition, motif analysis of enhancer-promoter interacting enhancers showed that GC-specific transcription factors are enriched. Among them, we found that MYB is crucial for GC cell growth and activated by the enhancer with an enhancer-promoter loop and TCF7  upregulation. Clinical GC samples showed epigenetic activation of enhancers at the MYB locus and significant upregulation of TCF7 and MYB , regardless of molecular GC subtype and clinicopathological factors. Single-cell RNA sequencing of gastric mucosa with intestinal metaplasia showed high expression of TCF7 and MYB in intestinal stem cells. When we inactivated the loop-forming enhancer at the MYB locus using CRISPR interference (dCas9-KRAB), GC cell growth was significantly inhibited. In conclusion, we identified MYB as an oncogene activated by a loop-forming enhancer and contributing to GC cell growth., (© The Author(s) 2024. Published by Oxford University Press on behalf of NAR Cancer.)

Objective: This study aimed to compare the stability and mechanical properties of the double chevron-cut (DCC) and biplanar (BP) distal femoral osteotomy (DFO) techniques, along with analyzing their respective contact surface areas., Methods: Biomechanical testing was performed using sawbone and 3D modeling techniques to assess axial and torsional stability, torsional stiffness, and maximum torque of both osteotomy configurations. Additionally, 3D models of the sawbone femur were created to calculate and compare the contact surface area of the DCC, BP, and conventional single-plane DFO techniques., Results: Axial stiffness and maximum strength did not significantly differ between the two osteotomy techniques. However, in terms of torsional properties, the DCC technique exhibited superior torsional stiffness compared to the BP group (27 ± 7.7 Nm/° vs. 4.5 ± 1.5 Nm/°, p = 0.008). Although the difference in maximum torque did not reach statistical significance (63 ± 10.6 vs. 56 ± 12.1, p = 0.87), it is noteworthy that the DCC group sawbone model exhibited fracture in the shaft region instead of at the osteotomy site. Therefore, the actual maximum torque of the DCC construct may not be accurately reflected by the numerical values obtained in this study. The contact surface area analysis revealed that the BP configuration had the largest contact surface area, 111% larger than that of the single-plane configuration. but 60% of it relied on the less reliable axial cut. Conversely, the DCC osteotomy offered a 31% larger contact surface area than the single-plane configuration, with both surfaces being weight-bearing., Conclusion: The DCC osteotomy exhibited superior mechanical stability, showing improved rotational stiffness and maximum torque when compared to the BP osteotomy. Although the BP osteotomy resulted in a larger contact surface area than the DCC osteotomy, both were larger than the conventional single-plane configuration. In clinical practice, both the DCC and BP techniques should be evaluated based on patient-specific characteristics and surgical goals., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Chen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: Fewer than 40% of U.S. children complete the human papillomavirus (HPV) vaccine series before their 13th birthday. In our large pediatric primary care network, HPV vaccine completion rate by age 13 was 30%. We hypothesized that a phased quality improvement (QI) initiative would increase rates of HPV vaccine completion by age 13 across our network., Methods: This QI initiative was conducted in a network of 30 practices located across two states, in urban and suburban settings, consisting of teaching and non-teaching clinics, and ranging in size from three to 50 providers per office. We used a phased approach incorporating multicomponent network-wide and iterative practice-specific interventions. Key interventions included: updating clinical decision support to default order HPV vaccine due at preventive visits starting at age nine instead of 11, data audit and feedback to providers and practices, encouraging use of a strong provider recommendation, and standing orders., Results: From April 2019 to October 2022, HPV vaccine completion by age 13 across our network increased from 30% to 55% and met criteria for special cause variation on statistical process control charts. A gap in median HPV vaccine completion by age 13 between patients with public insurance and patients with private or commercial insurance decreased from 9% to 1%., Conclusion: A QI initiative was associated with a sustained increase in HPV vaccine series completion by age 13 and reduced variation in care across a large network of 30 primary care practices., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Dr. Huang received an honorarium to present this work at the 26th Annual Pennsylvania Immunization Conference (June 2022)., (Copyright © 2024 Academic Pediatric Association. Published by Elsevier Inc. All rights reserved.)

Leveraging the specificity of antibody to deliver cytotoxic agent into tumor, antibody-drug conjugates (ADCs) have become one of the hotspots in the development of anticancer therapies. Although significant progress has been achieved, there remain challenges to overcome, including limited penetration into solid tumors and potential immunogenicity. Fully human single-domain antibodies (UdAbs), with their small size and human nature, represent a promising approach for addressing these challenges. Carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5) is a glycosylated cell surface protein that rarely expressed in normal adult tissues but overexpressed in diverse cancers, taking part in tumorigenesis, progression, and metastasis. In this study, we investigated the therapeutic potential of UdADC targeting CEACAM5. We performed biopanning in our library and obtained an antibody candidate B9, which bound potently and specifically to CEACAM5 protein (K D  = 4.84 nM) and possessed excellent biophysical properties (low aggregation tendency, high homogeneity, and thermal stability). The conjugation of B9 with a potent cytotoxic agent, monomethyl auristatin E (MMAE), exhibited superior antitumor efficacy against CEACAM5-expressing human gastric cancer cell line MKN-45, human pancreatic carcinoma cell line BxPC-3 and human colorectal cancer cell line LS174T with IC 50 values of 38.14, 25.60, and 101.4 nM, respectively. In BxPC-3 and MKN-45 xenograft mice, administration of UdADC B9-MMAE (5 mg/kg, i.v.) every 2 days for 4 times markedly inhibited the tumor growth without significant change in body weight. This study may have significant implications for the design of next-generation ADCs., (© 2023. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.)

Intestinal metaplasia (IM) is a pre-malignant condition of the gastric mucosa associated with increased gastric cancer (GC) risk. Analyzing 1,256 gastric samples (1,152 IMs) across 692 subjects from a prospective 10-year study, we identify 26 IM driver genes in diverse pathways including chromatin regulation (ARID1A) and intestinal homeostasis (SOX9). Single-cell and spatial profiles highlight changes in tissue ecology and IM lineage heterogeneity, including an intestinal stem-cell dominant cellular compartment linked to early malignancy. Expanded transcriptome profiling reveals expression-based molecular subtypes of IM associated with incomplete histology, antral/intestinal cell types, ARID1A mutations, inflammation, and microbial communities normally associated with the healthy oral tract. We demonstrate that combined clinical-genomic models outperform clinical-only models in predicting IMs likely to transform to GC. By highlighting strategies for accurately identifying IM patients at high GC risk and a role for microbial dysbiosis in IM progression, our results raise opportunities for GC precision prevention and interception., Competing Interests: Declaration of interests P.T. has stock in Tempus Healthcare, previous funding from Kyowa Hakko Kirin and Thermo Fisher Scientific, and patents/other intellectual property through the Agency for Science and Technology Research, Singapore (all outside the submitted work). K.G.Y. is a co-inventor on patents “Serum MicroRNA Biomarker for the Diagnosis of Gastric Cancer” and “Methods Related to Real-Time Cancer Diagnostics at Endoscopy Utilizing Fiber-Optic Raman Spectroscopy”; a member of Scientific Advisory Board of MiRXES Pte Ltd. He has no stock or shares in the related companies. He has no conflicts of interest to disclose regarding this submitted work. R.S. has received honoraria from MSD, Eli Lilly, BMS, Roche, Taiho, Astra Zeneca, DKSH, and Ipsen; has advisory activity with Bristol Myers Squibb, Merck, Eisai, Bayer, Taiho, Novartis, MSD, GSK, DKSH, and Astellas; received research funding from Paxman Coolers, MSD, and Natera; and has received travel grants from Roche, Astra Zeneca, Taiho, Eisai, and DKSH., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: DNA methylation accumulates in non-malignant gastric mucosa after exposure to pathogens. To elucidate how environmental, methylation, and lifestyle factors interplay to influence primary gastric neoplasia (GN) risk, we analyzed longitudinally monitored cohorts in Japan and Singapore., Methods: Asymptomatic subjects who underwent a gastric mucosal biopsy on the health check-up were enrolled. We analyzed the association between clinical factors and GN development using Cox hazard models. We further conducted comprehensive methylation analysis on selected tissues, including (i) mucosae from subjects developing GN later, (ii) mucosae from subjects not developing GN later, and (iii) GN tissues and surrounding mucosae. We also use the methylation data of mucosa collected in Singapore. The association between methylation and GN risk, as well as lifestyle and methylation, were analyzed., Findings: Among 4234 subjects, GN was developed in 77 subjects. GN incidence was correlated with age, drinking, smoking, and Helicobacter pylori (HP) status. Accumulation of methylation in biopsied gastric mucosae was predictive of higher future GN risk and shorter duration to GN incidence. Whereas methylation levels were associated with HP positivity, lifestyle, and morphological alterations, DNA methylation remained an independent GN risk factor through multivariable analyses. Pro-carcinogenic epigenetic alterations initiated by HP exposure were amplified by unfavorable but modifiable lifestyle choices. Adding DNA methylation to the model with clinical factors improved the predictive ability for the GN risk., Interpretation: The integration of environmental, lifestyle, and epigenetic information can provide increased resolution in the stratification of primary GN risk., Funding: The funds are listed in Acknowledgements section., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

Background and Aims: Peptic ulcer recurrent bleeding occurs in 20% to 30% of patients after standard endoscopic hemostasis, particularly within 4 days after the procedure. The application of additional tranexamic acid (TXA) to the ulcer may enhance hemostasis. This study investigated the effectiveness of TXA powder application on bleeding ulcers during endoscopic hemostasis., Methods: This study enrolled patients who had peptic ulcer bleeding between March 2022 and February 2023. After undergoing standard endoscopic therapy, the patients were randomly assigned to either the TXA group or the standard group. In the TXA group, an additional 1.25 g of TXA powder was sprayed endoscopically on the ulcer. Both groups then received 3 days of high-dose (8 mg/h) continuous infusion proton pump inhibitor therapy. Second-look endoscopy was conducted on days 3 to 4. The primary end point of early treatment failure was defined as ulcer recurrent bleeding within 4 days or major stigmata of recent hemorrhage on the second-look endoscopy., Results: Sixty patients (30 in each group) with peptic ulcer bleeding and balanced baseline characteristics were randomly assigned to a treatment group. The early treatment failure rate was lower in the TXA group (6.7%) than in the standard group (30%) (P = .042). The freedom from treatment failure periods for 4 and 28 days was significantly longer in the TXA group than in the standard group (P = .023). No adverse events from TXA were recorded., Conclusions: The precise delivery of topical TXA alongside standard endoscopic hemostasis reduced the early treatment failure rate in patients with bleeding peptic ulcers. (Clinical trial registration number: NCT05248321.)., Competing Interests: Disclosure All authors disclosed no financial relationships. This study was supported by the National Science and Technology Council, Taiwan (NSTC 111-2622-B-006-002) and National Cheng Kung University Hospital, Taiwan (NCKUH-11103012)., (Copyright © 2023 American Society for Gastrointestinal Endoscopy. Published by Elsevier Inc. All rights reserved.)

Objective: The long-term results of cervical disc arthroplasty (CDA) for noncontiguous cervical degenerative disc disease (CDDD) are still uncertain. Moreover, it is unclear whether CDA delays or avoids the degeneration of the intermediate segment (IS), leading to controversy in the field. Therefore, this study aimed to investigate the mid- to long-term clinical and radiographic outcomes of CDA in treating noncontiguous CDDD and to explore whether the IS degenerated faster after CDA than other non-surgically treated adjacent segments., Methods: We retrospectively analyzed patients with noncontiguous CDDD who underwent CDA in our department between January 2008 and July 2018. The patients were divided into the CDA and hybrid surgery (HS) groups, and clinical and radiographic outcomes were evaluated at routine postoperative intervals. Clinical outcomes were assessed using the Japanese Orthopaedic Association (JOA), neck disability index (NDI), and visual analogue scale (VAS), while radiographic outcomes included cervical lordosis (CL), C2-C7 range of motion (ROM), segmental ROM, and disc angle (DA) at the arthroplasty level. Complications were also evaluated.Pre- and postoperative values were compared using paired t-tests or Wilcoxon rank-sum tests. Independent Student t-tests or Mann-Whitney U tests analyzed continuous data between CDA and HS groups, while chi-square or Fisher exact tests assessed categorical data., Results: Sixty-four patients with noncontiguous CDDD, with 31 in the CDA group and 33 in the HS group, were evaluated. The mean follow-up time was over 70 months. The most frequently involved levels were C4/5 and C5/6. Both groups showed significant improvements in JOA, NDI, and VAS values after surgery. Although CL was maintained, the CL in the CDA group was consistently lower than that in the HS group (p < 0.05). There was a significant decrease in C2-C7 ROM (p < 0.05), but at the last follow-up, the C2-C7 ROM in the CDA group was greater than that in the HS group (p < 0.05). At the last follow-up, 44.3% of arthroplasty levels had developed heterotopic ossification (HO), and 48.45% had developed anterior bone loss (ABL). In addition, adjacent segment degeneration (ASDeg) was observed in the IS (22.7%), superior adjacent segment (20.6%)and inferior adjacent segment (21.9%)., Conclusion: CDA or CDA combined with fusion are viable treatments for noncontiguous CDDD, with satisfactory outcomes after mid-to-long-term follow-up. ASDeg is similar in non-surgical segments after 70 months of follow-up. ROM of the IS issimilar to preoperative levels, indicating CDA does not increase the risk of IS degeneration., (© 2023 The Authors. Orthopaedic Surgery published by Tianjin Hospital and John Wiley & Sons Australia, Ltd.)

Objective: Gastric cancer (GC) is a leading cause of cancer mortality, with ARID1A being the second most frequently mutated driver gene in GC. We sought to decipher ARID1A -specific GC regulatory networks and examine therapeutic vulnerabilities arising from ARID1A loss., Design: Genomic profiling of GC patients including a Singapore cohort (>200 patients) was performed to derive mutational signatures of ARID1A inactivation across molecular subtypes. Single-cell transcriptomic profiles of ARID1A -mutated GCs were analysed to examine tumour microenvironmental changes arising from ARID1A loss. Genome-wide ARID1A binding and chromatin profiles (H3K27ac, H3K4me3, H3K4me1, ATAC-seq) were generated to identify gastric-specific epigenetic landscapes regulated by ARID1A. Distinct cancer hallmarks of ARID1A -mutated GCs were converged at the genomic, single-cell and epigenomic level, and targeted by pharmacological inhibition., Results: We observed prevalent ARID1A inactivation across GC molecular subtypes, with distinct mutational signatures and linked to a NFKB-driven proinflammatory tumour microenvironment. ARID1A -depletion caused loss of H3K27ac activation signals at ARID1A -occupied distal enhancers, but unexpectedly gain of H3K27ac at ARID1A-occupied promoters in genes such as NFKB1 and NFKB2 . Promoter activation in ARID1A -mutated GCs was associated with enhanced gene expression, increased BRD4 binding, and reduced HDAC1 and CTCF occupancy. Combined targeting of promoter activation and tumour inflammation via bromodomain and NFKB inhibitors confirmed therapeutic synergy specific to ARID1A -genomic status., Conclusion: Our results suggest a therapeutic strategy for ARID1A -mutated GCs targeting both tumour-intrinsic (BRD4-assocatiated promoter activation) and extrinsic (NFKB immunomodulation) cancer phenotypes., Competing Interests: Competing interests: PT has stock and other ownership interests in Tempus Healthcare, previous research funding from Kyowa Hakko Kirin and Thermo Fisher Scientific, and patents/other intellectual property through the Agency for Science and Technology Research, Singapore (all outside the submitted work). RS has received honoraria from Bristol-Myers Squibb, Lilly, Roche, Taiho, Astra Zeneca, DKSH and MSD; has advisory activity with Bristol-Myers Squibb, Merck, Eisai, Bayer, Taiho, Novartis, MSD and AstraZeneca; received research funding from MSD and Paxman Coolers; and has received travel grants from AstraZeneca, Eisai, Roche and Taiho Pharmaceutical., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)

Chronic liver injury leads to progressive liver fibrosis and ultimately cirrhosis, a major cause of morbidity and mortality worldwide. However, there are currently no effective anti-fibrotic therapies available, especially for late-stage patients, which is partly attributed to the major knowledge gap regarding liver cell heterogeneity and cell-specific responses in different fibrosis stages. To reveal the multicellular networks regulating mammalian liver fibrosis from mild to severe phenotypes, we generated a single-nucleus transcriptomic atlas encompassing 49 919 nuclei corresponding to all main liver cell types at different stages of murine carbon tetrachloride (CCl 4 )-induced progressive liver fibrosis. Integrative analysis distinguished the sequential responses to injury of hepatocytes, hepatic stellate cells and endothelial cells. Moreover, we reconstructed cell-cell interactions and gene regulatory networks implicated in these processes. These integrative analyses uncovered previously overlooked aspects of hepatocyte proliferation exhaustion and disrupted pericentral metabolic functions, dysfunction for clearance by apoptosis of activated hepatic stellate cells, accumulation of pro-fibrotic signals, and the switch from an anti-angiogenic to a pro-angiogenic program during CCl 4 -induced progressive liver fibrosis. Our dataset thus constitutes a useful resource for understanding the molecular basis of progressive liver fibrosis using a relevant animal model.

Objective: Gastric cancer (GC) comprises multiple molecular subtypes. Recent studies have highlighted mesenchymal-subtype GC (Mes-GC) as a clinically aggressive subtype with few treatment options. Combining multiple studies, we derived and applied a consensus Mes-GC classifier to define the Mes-GC enhancer landscape revealing disease vulnerabilities., Design: Transcriptomic profiles of ~1000 primary GCs and cell lines were analysed to derive a consensus Mes-GC classifier. Clinical and genomic associations were performed across >1200 patients with GC. Genome-wide epigenomic profiles (H3K27ac, H3K4me1 and assay for transposase-accessible chromatin with sequencing (ATAC-seq)) of 49 primary GCs and GC cell lines were generated to identify Mes-GC-specific enhancer landscapes. Upstream regulators and downstream targets of Mes-GC enhancers were interrogated using chromatin immunoprecipitation followed by sequencing (ChIP-seq), RNA sequencing, CRISPR/Cas9 editing, functional assays and pharmacological inhibition., Results: We identified and validated a 993-gene cancer-cell intrinsic Mes-GC classifier applicable to retrospective cohorts or prospective single samples. Multicohort analysis of Mes-GCs confirmed associations with poor patient survival, therapy resistance and few targetable genomic alterations. Analysis of enhancer profiles revealed a distinctive Mes-GC epigenomic landscape, with TEAD1 as a master regulator of Mes-GC enhancers and Mes-GCs exhibiting preferential sensitivity to TEAD1 pharmacological inhibition. Analysis of Mes-GC super-enhancers also highlighted NUAK1 kinase as a downstream target, with synergistic effects observed between NUAK1 inhibition and cisplatin treatment., Conclusion: Our results establish a consensus Mes-GC classifier applicable to multiple transcriptomic scenarios. Mes-GCs exhibit a distinct epigenomic landscape, and TEAD1 inhibition and combinatorial NUAK1 inhibition/cisplatin may represent potential targetable options., Competing Interests: Competing interests: PT has stock and other ownership interests in Tempus Healthcare, previous research funding from Kyowa Hakko Kirin and Thermo Fisher Scientific, and patents/other intellectual property through the Agency for Science and Technology Research, Singapore (all outside the submitted work). RS has received honoraria from MSD, Eli Lilly, Bristol Myers Squibb, Roche, Taiho, Astra Zeneca and DKSH; has advisory activity with Bristol Myers Squibb, Merck, Eisai, Bayer, Taiho, Novartis, MSD and GSK; received research funding from MSD and Paxman Coolers; and has received travel grants from Astra Zeneca, Eisai, Roche, Taiho and DKSH. All remaining authors have declared no conflicts of interest., (© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.)

Nowadays, many old analog gauges still require the use of manual gauge reading. It is a time-consuming, expensive, and error-prone process. A cost-effective solution for automatic gauge reading has become a very important research topic. Traditionally, different types of gauges have their own specific methods for gauge reading. This paper presents a systematized solution called SGR (Scale-mark-based Gauge Reading) to automatically read gauge values from different types of gauges. Since most gauges have scale marks (circular or in an arc), our SGR algorithm utilizes PCA (principal components analysis) to find the primary eigenvector of each scale mark. The intersection of these eigenvectors is extracted as the gauge center to ascertain the scale marks. Then, the endpoint of the gauge pointer is found to calculate the corresponding angles to the gauge's center. Using OCR (optical character recognition), the corresponding dial values can be extracted to match with their scale marks. Finally, the gauge reading value is obtained by using the linear interpolation of these angles. Our experiments use four videos in real environments with light and perspective distortions. The gauges in the video are first detected by YOLOv4 and the detected regions are clipped as the input images. The obtained results show that SGR can automatically and successfully read gauge values. The average error of SGR is nearly 0.1% for the normal environment. When the environment becomes abnormal with respect to light and perspective distortions, the average error of SGR is still less than 0.5%.

For a broad range of applications, hyperspectral image (HSI) classification is a hot topic in remote sensing, and convolutional neural network (CNN)-based methods are drawing increasing attention. However, to train millions of parameters in CNN requires a large number of labeled training samples, which are difficult to collect. A conventional Gabor filter can effectively extract spatial information with different scales and orientations without training, but it may be missing some important discriminative information. In this article, we propose the Gabor ensemble filter (GEF), a new convolutional filter to extract deep features for HSI with fewer trainable parameters. GEF filters each input channel by some fixed Gabor filters and learnable filters simultaneously, then reduces the dimensions by some learnable 1×1 filters to generate the output channels. The fixed Gabor filters can extract common features with different scales and orientations, while the learnable filters can learn some complementary features that Gabor filters cannot extract. Based on GEF, we design a network architecture for HSI classification, which extracts deep features and can learn from limited training samples. In order to simultaneously learn more discriminative features and an end-to-end system, we propose to introduce the local discriminant structure for cross-entropy loss by combining the triplet hard loss. Results of experiments on three HSI datasets show that the proposed method has significantly higher classification accuracy than other state-of-the-art methods. Moreover, the proposed method is speedy for both training and testing.

Mutations in the DNA mismatch repair gene MSH2 are causative of microsatellite instability (MSI) in multiple cancers. Here, we discovered that besides its well-established role in DNA repair, MSH2 exerts a novel epigenomic function in gastric cancer. Unbiased CRISPR-based mass spectrometry combined with genome-wide CRISPR functional screening revealed that in early-stage gastric cancer MSH2 genomic binding is not randomly distributed but rather is associated specifically with tumor-associated super-enhancers controlling the expression of cell adhesion genes. At these loci, MSH2 genomic binding was required for chromatin rewiring, de novo enhancer-promoter interactions, maintenance of histone acetylation levels, and regulation of cell adhesion pathway expression. The chromatin function of MSH2 was independent of its DNA repair catalytic activity but required MSH6, another DNA repair gene, and recruitment to gene loci by the SWI/SNF chromatin remodeler SMARCA4/BRG1. Loss of MSH2 in advanced gastric cancers was accompanied by deficient cell adhesion pathway expression, epithelial-mesenchymal transition, and enhanced tumorigenesis in vitro and in vivo. However, MSH2-deficient gastric cancers also displayed addiction to BAZ1B, a bromodomain-containing family member, and consequent synthetic lethality to bromodomain and extraterminal motif (BET) inhibition. Our results reveal a role for MSH2 in gastric cancer epigenomic regulation and identify BET inhibition as a potential therapy in MSH2-deficient gastric malignancies., Significance: DNA repair protein MSH2 binds and regulates cell adhesion genes by enabling enhancer-promoter interactions, and loss of MSH2 causes deficient cell adhesion and bromodomain and extraterminal motif inhibitor synthetic lethality in gastric cancer., (©2022 American Association for Cancer Research.)

Purpose: To investigate the relationship between the preoperative paraspinal Goutalier grade of fatty infiltration and postoperative cervical sagittal alignment in patients undergoing anterior cervical discectomy and fusion (ACDF)., Methods: A total of 101 patients who underwent single-level ACDF with the Zero-profile implant system between March 2011 and April 2020 were included in this study. Cervical sagittal alignment parameters, including the C2-C7 Cobb angle, functional spinal unit (FSU) angle, cervical sagittal vertical axis (SVA), and T1 slope (T1S), were assessed. Preoperative magnetic resonance images were used to classify patients according to Goutalier grade. Clinical outcomes including Neck Disability Index (NDI) scores, Japanese Orthepaedic Association (JOA) scores and Visual Analogue Scale (VAS) scores were collected and analyzed., Results: According to the Goutalier grade, 33 patients were classified as Goutalier 0-1 (Group A), 44 were classified as Goutalier 1.5-2 (Group B), and 24 were classified as Goutalier 2.5-4.0 (Group C). The mean age among the three groups showed significant differences (P = 0.007). At the last follow-up, the C2-C7 Cobb angle, FSU angle, and T1S improved after the surgery among the groups. Although there were varying degrees of loss of curvature among the different groups during the follow-up period, the postoperative cervical sagittal alignment parameters demonstrated no statistical differences among the three groups (P > 0.05). In addition, patients in all groups experienced significant relief of their symptoms, and the clinical scores were comparable among the groups (P > 0.05)., Conclusion: The complex nature of anterior cervical surgery requires surgical attention both in decompression and sagittal alignment. Our study demonstrates satisfactory postoperative cervical sagittal alignment of patients despite different grades of fatty infiltration of the multifidus muscle following single-level ACDF. Based on our results, the improvement and maintenance of cervical sagittal alignment after ACDF remains a complex problem that spine surgeons should consider before surgery., (© 2022. The Author(s).)

Objectives: Epigenomic alterations in cancer interact with the immune microenvironment to dictate tumour evolution and therapeutic response. We aimed to study the regulation of the tumour immune microenvironment through epigenetic alternate promoter use in gastric cancer and to expand our findings to other gastrointestinal tumours., Design: Alternate promoter burden (APB) was quantified using a novel bioinformatic algorithm ( proActiv ) to infer promoter activity from short-read RNA sequencing and samples categorised into APB high , APB int and APB low. Single-cell RNA sequencing was performed to analyse the intratumour immune microenvironment. A humanised mouse cancer in vivo model was used to explore dynamic temporal interactions between tumour kinetics, alternate promoter usage and the human immune system. Multiple cohorts of gastrointestinal tumours treated with immunotherapy were assessed for correlation between APB and treatment outcomes., Results: APB high gastric cancer tumours expressed decreased levels of T-cell cytolytic activity and exhibited signatures of immune depletion. Single-cell RNAsequencing analysis confirmed distinct immunological populations and lower T-cell proportions in APB high tumours. Functional in vivo studies using 'humanised mice' harbouring an active human immune system revealed distinct temporal relationships between APB and tumour growth, with APB high tumours having almost no human T-cell infiltration. Analysis of immunotherapy-treated patients with GI cancer confirmed resistance of APB high tumours to immune checkpoint inhibition. APB high gastric cancer exhibited significantly poorer progression-free survival compared with APB low (median 55 days vs 121 days, HR 0.40, 95% CI 0.18 to 0.93, p=0.032)., Conclusion: These findings demonstrate an association between alternate promoter use and the tumour microenvironment, leading to immune evasion and immunotherapy resistance., Competing Interests: Competing interests: The subject matter in this manuscript was submitted as a technology disclosure to the institutional Technology Transfer Office for potential intellectual property protection. PT had stock and other ownership interests in HealthSeq, research funding from Kyowa Hakko Kirin and Thermo Fisher Scientific, and patents/other intellectual property through the Agency for Science and Technology Research, Singapore (all outside the submitted work). RS received honoraria from Bristol-Myers Squibb, Lilly, Roche, Taiho, Astra Zeneca, DKSH and MSD; had advisory activity with Bristol-Myers Squibb, Eisai, Merck, Bayer, Taiho, Novartis, MSD and AstraZeneca; received research funding from Paxman Coolers and MSD; received travel grants from AstraZeneca, Roche, Eisai and Taiho Pharmaceutical (all outside the submitted work). FP received honoraria for speakers’ bureau/advisory activity from Amgen, Merck-Serono, Roche, Lilly, Sanofi, Bayer and Servier, and received research funding from BMS (all outside the submitted work)., (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Coronary artery disease (CAD) is a multifactorial disease and its pathogenesis remains unclear. We aimed to explore the optimal feature genes (OFGs) for CAD and to investigate the function of immune cell infiltration of CAD. It will be helpful for better understanding of the pathogenesis and the development of genetic prediction of CAD., Methods: Datasets related to CAD were obtained from the Gene Expression Omnibus (GEO) database. Cases from the datasets met diagnostic criteria including clinical symptoms, electrocardiographic (ECG) and angiographic evidence. We identified differentially expressed genes (DEGs) and conducted functional enrichment analysis. OFGs were obtained from the least absolute shrinkage and selection operator (LASSO) algorithm, support vector machine recursive feature elimination (SVM-RFE) algorithm, and random forest (RF) algorithm. CIBERSORT was used to compare immune infiltration between CAD patients and normal controls, and the correlation between OFGs and immune cells was analyzed., Results: DEGs were involved in the interleukin (IL)-17 signaling pathway, nuclear factor (NF)-kappa B signaling pathway, and tumor necrosis factor (TNF) signaling pathway. Gene Ontology (GO) analysis revealed DEGs were enriched in lipopolysaccharide (LPS), tertiary granule, and pattern recognition receptor activity. Disease Ontology (DO) analysis suggested DEGs were enriched in lung disease, arteriosclerotic cardiovascular disease (CVD). Matrix metalloproteinase 9 (MMP9), Pellino E3 ubiquitin protein ligase 1 (PELI1), thrombomodulin (THBD), and zinc finger protein 36 (ZFP36) were screened by the intersection of OFGs obtained from LASSO, SVM-REF, and RF algorithms. CAD patients had a lower proportion of memory B cells (P=0.019), CD8 T cells (P<0.001), resting memory CD4 T cells (P<0.001), regulatory T cells (P=0.028), and gamma delta T cells (P<0.001) than normal controls, while the proportion of activated memory CD4 T cells (P=0.014), resting natural killer (NK) cells (P<0.001), monocytes (P<0.001), M0 macrophages (P=0.023), activated mast cells (P<0.001), and neutrophils (P<0.001) in CAD patients were higher than normal controls. MMP9, PELI1, THBD, and ZFP36 were correlated with immune cells., Conclusions: MMP9, PELI1, THBD, and ZFP36 may be predicted biomarkers for CAD. The OFGs and association between OFGs and immune infiltration may provide potential biomarkers for CAD prediction along with the better assessment of the disease., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://jtd.amegroups.com/article/view/10.21037/jtd-22-632/coif). The authors have no conflicts of interest to declare., (2022 Journal of Thoracic Disease. All rights reserved.)

This study aimed to investigate clinical effectiveness of stone disintegration by using isolation coupling pad ("icPad") as coupling medium to reduce trapped air pockets during extracorporeal shock wave lithotripsy (ESWL). Patients underwent ESWL between Oct. 2017 and May 2018 were enrolled in this clinical observational study. An electromagnetic lithotripter (Dornier MedTech Europe GmbH Co., Germany) was used in this study. Patients were divided into icPad group P1, P2 and semi-gel group C by different coupling medium. The energy level and total number of shock wave (SW) for group P1 and C was set at level 2 and 3000 and group P2 at level 3 and 2500. The successful stone disintegration rate (SSDR) was determined to evaluate the treatment outcome. All patients were evaluated by KUB film and ultrasonography after 90 days. Complications during ESWL were recorded. A total of 300 patients satisfied the inclusion criteria. There were no significant differences in characteristics of patients and stone among three groups. The corresponding SSDRs for patients in group P1, P2 and C was 73.0%, 73.2% and 55.3%, respectively. The SSDR in group P1 was statistically higher than Group C. Comparing to semi-liquid gel, coupling medium using by icPad could achieve better treatment outcome of stone disintegration in ESWL., (© 2022. The Author(s).)

Gastric cancer heterogeneity represents a barrier to disease management. We generated a comprehensive single-cell atlas of gastric cancer (>200,000 cells) comprising 48 samples from 31 patients across clinical stages and histologic subtypes. We identified 34 distinct cell-lineage states including novel rare cell populations. Many lineage states exhibited distinct cancer-associated expression profiles, individually contributing to a combined tumor-wide molecular collage. We observed increased plasma cell proportions in diffuse-type tumors associated with epithelial-resident KLF2 and stage-wise accrual of cancer-associated fibroblast subpopulations marked by high INHBA and FAP coexpression. Single-cell comparisons between patient-derived organoids (PDO) and primary tumors highlighted inter- and intralineage similarities and differences, demarcating molecular boundaries of PDOs as experimental models. We complemented these findings by spatial transcriptomics, orthogonal validation in independent bulk RNA-sequencing cohorts, and functional demonstration using in vitro and in vivo models. Our results provide a high-resolution molecular resource of intra- and interpatient lineage states across distinct gastric cancer subtypes., Significance: We profiled gastric malignancies at single-cell resolution and identified increased plasma cell proportions as a novel feature of diffuse-type tumors. We also uncovered distinct cancer-associated fibroblast subtypes with INHBA-FAP-high cell populations as predictors of poor clinical prognosis. Our findings highlight potential origins of deregulated cell states in the gastric tumor ecosystem. This article is highlighted in the In This Issue feature, p. 587., (©2021 The Authors; Published by the American Association for Cancer Research.)

Psoriasis is an autoimmune disease characterized by chronic skin inflammation and excessive keratinocyte proliferation. The itchy, scaly, and erythematous lesions present on psoriatic skin negatively affect patients' quality of life. Pinocembrin is a flavonoid present in propolis, fruits, and vegetables. It exerts neuroprotective effects and was used for treating ischemic stroke in a human clinical trial. However, the effects of pinocembrin on psoriasis have never been examined. In this study, we evaluated the effects of pinocembrin on human HaCaT keratinocytes and BALB/c mice with imiquimod- (IMQ-) induced psoriatic dermatitis. In interferon- γ - (IFN- γ -) activated HaCaT cells, pinocembrin reduced the expression of inflammatory cytokines, namely, tumor necrosis factor- α (TNF- α ), interleukin-6 (IL-6), and keratinocyte proliferation markers, namely, keratin (K)16, K17, and Ki-67. The mechanism underlying these inhibitory effects involved the regulation of the heme oxygenase- (HO-) 1/signal transducer and activator of transcription (STAT) 3 pathway. In the IMQ-induced psoriatic dermatitis mouse model, the topical application of pinocembrin significantly ameliorated the Skin Psoriasis Area and Severity Index score, epidermal thickness, inflammation, hyperplasia, hyperkeratosis, and cluster of differentiation (CD) 4 + T-cell infiltration. Expression of the inflammatory cytokines and keratinocyte proliferation markers in dorsal skin was significantly decreased in the pinocembrin-treated group. Meanwhile, in lesional skin, the expression of HO-1 was upregulated, but that of phospho-STAT3 (pSTAT3) was downregulated. Collectively, our results indicated the therapeutic potential of pinocembrin. Additional studies are warranted to evaluate its clinical benefits in patients with psoriasis., Competing Interests: The authors declare no conflicts of interest regarding the publication of this paper., (Copyright © 2022 Kuo-Kuei Huang et al.)

Multi-source domain adaptation (MDA) aims to transfer knowledge from multiple source domains to an unlabeled target domain. MDA is a challenging task due to the severe domain shift, which not only exists between target and source but also exists among diverse sources. Prior studies on MDA either estimate a mixed distribution of source domains or combine multiple single-source models, but few of them delve into the relevant information among diverse source domains. For this reason, we propose a novel MDA approach, termed Pseudo Target for MDA (PTMDA). Specifically, PTMDA maps each group of source and target domains into a group-specific subspace using adversarial learning with a metric constraint, and constructs a series of pseudo target domains correspondingly. Then we align the remainder source domains with the pseudo target domain in the subspace efficiently, which allows to exploit additional structured source information through the training on pseudo target domain and improves the performance on the real target domain. Besides, to improve the transferability of deep neural networks (DNNs), we replace the traditional batch normalization layer with an effective matching normalization layer, which enforces alignments in latent layers of DNNs and thus gains further promotion. We give theoretical analysis showing that PTMDA as a whole can reduce the target error bound and leads to a better approximation of the target risk in MDA settings. Extensive experiments demonstrate PTMDA's effectiveness on MDA tasks, as it outperforms state-of-the-art methods in most experimental settings.

Background: Enhancers are distal cis-regulatory elements required for cell-specific gene expression and cell fate determination. In cancer, enhancer variation has been proposed as a major cause of inter-patient heterogeneity-however, most predicted enhancer regions remain to be functionally tested., Methods: We analyzed 132 epigenomic histone modification profiles of 18 primary gastric cancer (GC) samples, 18 normal gastric tissues, and 28 GC cell lines using Nano-ChIP-seq technology. We applied Capture-based Self-Transcribing Active Regulatory Region sequencing (CapSTARR-seq) to assess functional enhancer activity. An Activity-by-contact (ABC) model was employed to explore the effects of histone acetylation and CapSTARR-seq levels on enhancer-promoter interactions., Results: We report a comprehensive catalog of 75,730 recurrent predicted enhancers, the majority of which are GC-associated in vivo (> 50,000) and associated with lower somatic mutation rates inferred by whole-genome sequencing. Applying CapSTARR-seq to the enhancer catalog, we observed significant correlations between CapSTARR-seq functional activity and H3K27ac/H3K4me1 levels. Super-enhancer regions exhibited increased CapSTARR-seq signals compared to regular enhancers, even when decoupled from native chromatin contexture. We show that combining histone modification and CapSTARR-seq functional enhancer data improves the prediction of enhancer-promoter interactions and pinpointing of germline single nucleotide polymorphisms (SNPs), somatic copy number alterations (SCNAs), and trans-acting TFs involved in GC expression. We identified cancer-relevant genes (ING1, ARL4C) whose expression between patients is influenced by enhancer differences in genomic copy number and germline SNPs, and HNF4α as a master trans-acting factor associated with GC enhancer heterogeneity., Conclusions: Our results indicate that combining histone modification and functional assay data may provide a more accurate metric to assess enhancer activity than either platform individually, providing insights into the relative contribution of genetic (cis) and regulatory (trans) mechanisms to GC enhancer functional heterogeneity., (© 2021. The Author(s).)

Introduction: Genes contain multiple promoters that can drive the expression of various transcript isoforms. Although transcript isoforms from the same gene could have diverse and non-overlapping functions, current loss-of-function methodologies are not able to differentiate between isoform-specific phenotypes., Results: Here, we show that CRISPR interference (CRISPRi) can be adopted for targeting specific promoters within a gene, enabling isoform-specific loss-of-function genetic screens. We use this strategy to test functional dependencies of 820 transcript isoforms that are gained in gastric cancer (GC). We identify a subset of GC-gained transcript isoform dependencies, and of these, we validate CIT kinase as a novel GC dependency. We further show that some genes express isoforms with opposite functions. Specifically, we find that the tumour suppressor ZFHX3 expresses an isoform that has a paradoxical oncogenic role that correlates with poor patient outcome., Conclusions: Our work finds isoform-specific phenotypes that would not be identified using current loss-of-function approaches that are not designed to target specific transcript isoforms.

Background: Deregulated gene expression is a hallmark of cancer; however, most studies to date have analyzed short-read RNA sequencing data with inherent limitations. Here, we combine PacBio long-read isoform sequencing (Iso-Seq) and Illumina paired-end short-read RNA sequencing to comprehensively survey the transcriptome of gastric cancer (GC), a leading cause of global cancer mortality., Results: We performed full-length transcriptome analysis across 10 GC cell lines covering four major GC molecular subtypes (chromosomal unstable, Epstein-Barr positive, genome stable and microsatellite unstable). We identify 60,239 non-redundant full-length transcripts, of which > 66% are novel compared to current transcriptome databases. Novel isoforms are more likely to be cell line and subtype specific, expressed at lower levels with larger number of exons, with longer isoform/coding sequence lengths. Most novel isoforms utilize an alternate first exon, and compared to other alternative splicing categories, are expressed at higher levels and exhibit higher variability. Collectively, we observe alternate promoter usage in 25% of detected genes, with the majority (84.2%) of known/novel promoter pairs exhibiting potential changes in their coding sequences. Mapping these alternate promoters to TCGA GC samples, we identify several cancer-associated isoforms, including novel variants of oncogenes. Tumor-specific transcript isoforms tend to alter protein coding sequences to a larger extent than other isoforms. Analysis of outcome data suggests that novel isoforms may impart additional prognostic information., Conclusions: Our results provide a rich resource of full-length transcriptome data for deeper studies of GC and other gastrointestinal malignancies.

Objective: To explore the clinical application value of routine indicators such as blood routine and liver and kidney function in auxiliary diagnosis and prognosis of COVID-19 patients. Methods: SNK-q and other methods were used to retrospectively analyzed the differences of blood routine test, liver and kidney function and other inflammatory indexes of 30 patients with covid-19, 29 patients with other viral pneumonia, 35 patients with influenza A/B and 25 healthy persons from January 28 to February 14, 2020 in Xiangya Hospital of Central South University. Results: The neutrophils count increased gradually in COVID-19 group, influenza A/B group and other types of viral pneumonia group, and the difference between COVID-19 group and other viral pneumonia groups was statistically significant( H =-19.064, P< 0.05); The lymphocyte count decreased gradually in the control group, influenza A/B group, other viral pneumonia group and COVID-19 group. In addition, DB, UA and GLU were also different among groups. Subgroup analysis showed that there were statistically significant differences in N( F= 9.581, t= -0.152, P< 0.05), N%( F= 5.723, t= -0.600, P< 0.05), NLR( F= 4.773, t= -1.161, P< 0.05), PCT( F= 17.464, t= -1.477, P< 0.05)and CRP( F= 7.656, t= -1.973, P< 0.05) between patients with lung involvement +-++ and patients with lung involvement +++-++++. There were statistically significant differences in NLR( F= 63.931, t= -2.815, P< 0.01), AST( F= 15.704, t= -1.930, P< 0.01), ALT( F= 35.551, t= -2.199, P< 0.01), LDH( F= 7.715, t= -2.703, P< 0.05) and GLU( F= 6.306, t= -5.116, P< 0.05) between the light+common subgroup and the heavy+critical subgroup of COVID-19 clinical classification. Correlation analysis showed that clinical stage and imaging credit period were significantly correlated with NLR ( r =0.406 , P =0.026; r =0.397 , P =0.030), ALT ( r =0.403 , P =0.049; r =0.418 , P =0.047), LDH ( r =0.543 , P <0.01; r =0.643 , P <0.01) and GLU( r= 0.750 , P <0.01; r =0.471 , P =0.042). A total of 5 principal components were extracted from all the included indicators, and the comprehensive information extraction rate was 82.86%. Indicators of a large load included Ur, PCT and CRP in PC1; ALT, AST and GLU in PC2; N%, L%, L and NLR in PC3. It indicated that the indicators of acute infection, liver function and blood routine had certein warning effect on disease surveillance. The results of ROC curve analysis showed that the combined detection of N+TB+Urea was the best practice to distinguish COVID-19 and other viral pneumonia, while the combined detection of N+L+UA was the most effective solution to make a distinction between COVID-19 and influenza A/B patients. In the aspect of disease evaluation, NL+LDH+GLU+ALT combined detection represent the best diagnostic performance to distinguish the clinical stage of light+common type and heavy+critical type, achieving the AUC (ROC) to 0.904, with the sensitivity 75% and the specificity 100% at the cut-off value of 0.477. Conclusion: In addition to etiology and imaging examination, doctors can also improve the routine laboratory tests such as blood routine test, liver and kidney function to assist diagnosis and disease prediction of patients with respiratory tract infection.

Objective: To explore the relationship between the total cerebral small vessel disease (CSVD) score and retinal vessel diameters in patients with mild stroke. Methods: The patients with mild stroke who were hospitalized in the Second People's Hospital of Changzhou, Nanjing Medical University from March to December 2019 were continuously collected (National Institutes of Health Stroke Scale score≤3 points). All patients completed the head magnetic resonance imaging and retinal fundus photography examination, and then the retinal arteriovenous diameter was measured semi-automatically based on the pictures. According to the total CSVD score (0-4 points), the patients were divided into 5 groups. The baseline characteristics of the patients were compared. Moreover, the correlation of total CSVD with retinal blood vessel diameters were analyzed by spearman and linear regression. Results: A total of 206 patients were enrolled. There were 69, 51, 41, 30, and 15 patients with 0, 1, 2, 3, and 4 points, respectively. In CSVD subgroups, there were significant differences in age, duration of hypertension and diabetes (all P< 0.05). The central retinal artery equivalent (CRAE), (CSVD scores 0-4 were (126±12) μm, (118±11) μm, (108±11) μm, (99±8) μm, (90±7) μm, P <0.001) and arteriole-to-venule ratio (AVR) (CSVD scores 0-4 were 0.65±0.05, 0.60±0.04, 0.56±0.04, 0.49±0.03, 0.44±0.02, P <0.001) were different in CSVD subgroups. With the increase of CSVD score, the diameter of artery and AVR became smaller. The total CSVD was significantly correlated with AVR by Spearman correlation analysis ( r = 0.818, P <0.001). By constructing a linear regression equation model, the coefficient of determination of the total CSVD score ( R 2 =0.694) was higher than that of lacunes, white matter hyperintensities, cerebral microbleeds and enlarged perivascular space. After adjusting for age, course of hypertension and diabetes, and different types of CSVD, further multiple linear regression analysis revealed that the total CSVD score was still an independent related factor of AVR (β=-0.039, P <0.001, 95% CI =-0.051--0.028). Conclusions: Total CSVD score is negatively correlated with retinal artery diameters and AVR. Additionally, the total CSVD score can better reflect the degree of cerebral microvascular lesions than single type CSVD.

Objective: To investigate the effects of preoperative segmental range of motion (ROM) on clinical and radiographical outcomes after artificial cervical disc replacement (ACDR) and explore whether ACDR could be indicated for patients with preoperative limited or excessive segmental ROM. Methods: From January 2008 to December 2017, patients who underwent Prestige-LP ACDR in West China Hospital were retrospectively reviewed. The preoperative and postoperative X-rays of the cervical spine were collected to measure the radiographic parameters, including cervical lordosis (CL), C(2-7) ROM, disc height (DH), disc angle (DA) and ROM at the arthroplasty level. Clinical outcomes were evaluated using the Japanese Orthopedic Association (JOA) and the Neck Disability Index (NDI) scores. The correlation between preoperative segmental ROM and postoperative clinical and radiographical outcomes were also analyzed. Results: A total of 161 patients were analyzed, with 73 males and 88 females. The mean age was (44±8) years, and the follow-up period was 34 months (12-120 months). JOA and NDI scores improved after ACDR ( P< 0.05). However, postoperative C(2-7) ROM and ROM at the arthroplasty level were comparable with preoperative counterparts (both P> 0.05). Preoperative segmental ROM positively correlated with C(2-7) ROM and ROM at the arthroplasty segment ( r= 0.213、0.271, both P< 0.05), but was negatively correlated with the change of ROM ( r= -0.534, P< 0.05). The segmental ROM was 4.0°±1.0° in the limited-ROM group (A) and 14.6°±1.3° in the excessive-ROM group (B), respectively. There were significantly more patients diagnosed with cervical spondylosis in group A than in group B (35.5% vs 10.7%, P< 0.05). The level-distribution was statistically different between the two groups. C(5/6) and C(6/7) were prone to limited motion in group A, while C(4/5) and C(5/6) were predisposed to excessive motion in group B (all P< 0.05). After surgery, C(2-7) ROM increased for 14.2°±16.8° in group A, while paradoxically decreased for 2.2°±14.4° in group B. However, C(2-7) ROM in group B was still larger than that in group A ( P< 0.05). Similarly, the ROM at the arthroplasty level increased by 3.1°±3.7° in group A, whereas the values decreased by 4.4°±4.2° in group B postoperatively. In addition, group A still had less segmental ROM than group B ( P< 0.05). The preoperative DH in group A was less than that in group B ( P< 0.05). The rates of ASD, HO, and high-grade HO in group A were all higher than those in group B but without significant differences (all P> 0.05). Conclusion: Preoperative segmental ROM has no significant effects on clinical outcomes after ACDR; it has a positive correlation with postoperative global and segmental ROM while is negatively correlated with ROM change.

Objective: Intestinal metaplasia (IM) is a premalignant stage that poses a greater risk for subsequent gastric cancer (GC). However, factors regulating IM to GC progression remain unclear. Previously, activated DNA damage response (DDR) signalling factors were shown to engage tumour-suppressive networks in premalignant lesions. Here, we interrogate the relationship of DDR signalling to mutational accumulation in IM lesions., Design: IM biopsies were procured from the gastric cancer epidemiology programme, an endoscopic surveillance programme where biopsies have been subjected to (epi)genomic characterisation. IM samples were classified as genome-stable or genome-unstable based on their mutational burden/somatic copy-number alteration (CNA) profiles. Samples were probed for DDR signalling and cell proliferation, using the markers γH2AX and MCM2, respectively. The expression of the gastric stem cell marker, CD44v9, was also assessed. Tissue microarrays representing the GC progression spectrum were included., Results: MCM2-positivity increased during GC progression, while γH2AX-positivity showed modest increase from normal to gastritis and IM stages, with further increase in GC. γH2AX levels correlated with the extent of chronic inflammation. Interestingly, genome-stable IM lesions had higher γH2AX levels underscoring a protective anti-cancer role for DDR signalling. In contrast, genome-unstable IM lesions with higher mutational burden/CNAs had lower γH2AX levels, elevated CD44v9 expression and modest promoter hypermethylation of DNA repair genes WRN, MLH1 and RAD52 ., Conclusions: Our data suggest that IM lesions with active DDR will likely experience a longer latency at the premalignant state until additional hits that override DDR signalling clonally expand and promote progression. These observations provide insights on the factors governing IM progression., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Spasmolytic polypeptide-expressing metaplasia (SPEM) and pyloric gland adenoma (PGA) in the stomach are metaplastic and neoplastic lesions, respectively, in which gastric body glands are replaced by pyloric glands. The aim of this study was to evaluate the genomic profile of SPEM and compare it with intestinal-type gastric cancer (GC) and PGA. Thirteen gastrectomies showing PGA with or without dysplasia, GC and SPEM were retrospectively selected. MUC5AC, MUC6, gastrin, and TFF2 IHC were performed. Lesions were subjected to laser capture microdissection followed by DNA extraction. Forty-three DNA samples were extracted from PGA without cytological dysplasia, PGA with low-grade and high-grade dysplasia and pyloric gland adenocarcinoma, GC, SPEM, and adjacent normal tissue from the body of the stomach and were subjected to exome sequencing for 49 genes that are commonly dysregulated in GC. Sanger sequencing was performed for confirmation. Twenty nonsynonymous mutations were identified in SPEM, and none of these were frameshifts or indels. PGA with or without cytological dysplasia showed a significantly higher number of mutations compared with SPEM. As cytological dysplasia increased from no dysplasia to dysplasia in PGA, the percentage of frameshift mutations, indels, and missense variations increased. Further missense or frameshift mutations were observed in the KRAS, APC, TP53, and CTNNB1 genes in the PGA group. In GC, mutations were observed in the TP53 gene (p.Arg248Gln). Missense mutations in the MUC5AC, KRAS, BRAF, and EZH2 genes were common between SPEM and GC. SPEM showed fewer genomic variations than GC and PGA, and was genomically distinct from the pyloric epithelium in PGA. Stepwise progression of PGA from PGA without dysplasia to PGA with dysplasia/adenocarcinoma was associated an increase in mutations. SPEM appears to be more genomically similar to GC than PGA.

Epstein-Barr virus (EBV) is associated with several human malignancies including 8-10% of gastric cancers (GCs). Genome-wide analysis of 3D chromatin topologies across GC lines, primary tissue and normal gastric samples revealed chromatin domains specific to EBV-positive GC, exhibiting heterochromatin-to-euchromatin transitions and long-range human-viral interactions with non-integrated EBV episomes. EBV infection in vitro suffices to remodel chromatin topology and function at EBV-interacting host genomic loci, converting H3K9me3 + heterochromatin to H3K4me1 + /H3K27ac + bivalency and unleashing latent enhancers to engage and activate nearby GC-related genes (for example TGFBR2 and MZT1). Higher-order epigenotypes of EBV-positive GC thus signify a novel oncogenic paradigm whereby non-integrative viral genomes can directly alter host epigenetic landscapes ('enhancer infestation'), facilitating proto-oncogene activation and tumorigenesis.

Background Context: The development of bone loss (BL) at the operated level after cervical disc arthroplasty (CDA) has not been well recognized. The incidence of BL may be correlated with the prosthesis type. Currently, no study has reported the incidence of BL after CDA with the Prestige-LP disc, and this remains an active area of research., Purpose: To determine the incidence of BL after Prestige-LP CDA and evaluate the impact of BL on clinical and radiological outcomes., Study Design: This is an observational study., Patient Sample: A total of 396 patients were reviewed., Outcome Measures: The Japanese Orthopedics Association (JOA), Visual Analogue Scale (VAS), and Neck Disability Index (NDI) scores were evaluated. Cervical lordosis, disc angle, global and segmental range of motion (ROM), heterotopic ossification (HO), and BL were measured., Methods: We retrospectively reviewed patients who underwent Prestige-LP disc from January 2008 to October 2018 at our institution. Clinical outcomes were evaluated using JOA, VAS, and NDI scores. Radiological variables, including cervical lordosis, disc angle, global and segmental ROM, HO, and BL, were retrieved., Results: A total of 396 patients and 483 CDAs were evaluated. BL occurred in 56.6% of patients and 52.8% of CDA segments. Mild BL occurred in 30.2%, moderate BL in 37.3%, and severe BL in 32.5% of CDA segments. Notably, 88.2% of CDA segments developed BL within the first 3 months, and 19.1% of them progressed at 6 months. However, no progressive BL after 12 months was seen. About 50.2% of CDAs showed superior and inferior endplates involvement. The incidence of BL was associated with age, surgery type, level distribution, and incidence and grade of HO. Patients with BL had a better segmental ROM, but no relationships between patients with or without BL were found in clinical outcomes., Conclusions: BL was a common but self-limited phenomenon after CDA at the early postoperative stage. It occurred more often in relatively young age patients, two-level CDA, and C5/6 segment. However, patients suffering from BL showed no deterioration of the clinical outcomes, more exceptional motion preservation at the arthroplasty level, and lower incidence with a lower grade of HO., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Cervical disc arthroplasty (CDA) has been demonstrated, in clinical trials, as an effective and safe treatment for patients diagnosed with radiculopathy and/or myelopathy. However, the current CDA indication criteria, based on the preoperative segmental range of motion (ROM), comprises a wide range of variability. Although the arthroplasty level preserved ROM averages 7°-9° after CDA, there are no clear guidelines on preoperatively limited or excessive ROM at the index level, which could be considered as suitable for CDA., Methods: This was a retrospective study of patients who underwent CDA between January 2008 and October 2018 using Prestige-LP discs in our hospital. They were divided into the small-ROM (≤5.5°) and the large-ROM (> 12.5°) groups according to preoperatively index-level ROM. Clinical outcomes, including the Japanese Orthopedics Association (JOA), Neck Disability Index (NDI), and Visual Analogue Scale (VAS) scores, were evaluated. Radiological parameters, including cervical lordosis, disc angle (DA), global and segmental ROM, disc height (DH), and complications were measured., Results: One hundred and twenty six patients, with a total of 132 arthroplasty segments were analyzed. There were 64 patients in the small-ROM and 62 in the large-ROM group. There were more patients diagnosed with cervical spondylosis in the small-ROM than in the large-ROM group (P = 0.046). Patients in both groups had significantly improved JOA, NDI, and VAS scores after surgery, but the intergroup difference was not significant. Patients in the small-ROM group had dramatic postoperative increase in cervical lordosis, global and segmental ROM (P < 0.001). However, there was a paradoxical postoperative decrease in global and segmental ROM in the large-ROM group postoperatively (P < 0.001). Patients in the small-ROM group had lower preoperative DH (P = 0.012), and a higher rate of postoperative heterotopic ossification (HO) (P = 0.037)., Conclusion: Patients with preoperatively limited segmental ROM had severe HO, and achieved similar postoperative clinical outcomes as patients with preoperatively excessive segmental ROM. Patients with preoperatively limited segmental ROM showed a postoperative increase in segmental mobility, which decreased in patients with preoperatively excessive segmental ROM.

Transcriptional reactivation of telomerase catalytic subunit (TERT) is a frequent hallmark of cancer, occurring in 90% of human malignancies. However, specific mechanisms driving TERT reactivation remain obscure for many tumor types and in particular gastric cancer (GC), a leading cause of global cancer mortality. Here, through comprehensive genomic and epigenomic analysis of primary GCs and GC cell lines, we identified the transcription factor early B cell factor 1 (EBF1) as a TERT transcriptional repressor and inactivation of EBF1 function as a major cause of TERT upregulation. Abolishment of EBF1 function occurs through 3 distinct (epi)genomic mechanisms. First, EBF1 is epigenetically silenced via DNA methyltransferase, polycomb-repressive complex 2 (PRC2), and histone deacetylase activity in GCs. Second, recurrent, somatic, and heterozygous EBF1 DNA-binding domain mutations result in the production of dominant-negative EBF1 isoforms. Third, more rarely, genomic deletions and rearrangements proximal to the TERT promoter remobilize or abolish EBF1-binding sites, derepressing TERT and leading to high TERT expression. EBF1 is also functionally required for various malignant phenotypes in vitro and in vivo, highlighting its importance for GC development. These results indicate that multimodal genomic and epigenomic alterations underpin TERT reactivation in GC, converging on transcriptional repressors such as EBF1.

Objective: Gastric cancer (GC) is a leading cause of cancer mortality. Previous studies have shown that hepatocyte nuclear factor-4α (HNF4α) is specifically overexpressed in GC and functionally required for GC development. In this study, we investigated, on a genome-wide scale, target genes of HNF4α and oncogenic pathways driven by HNF4α and HNF4α target genes., Design: We performed HNF4α chromatin immunoprecipitation followed by sequencing across multiple GC cell lines, integrating HNF4α occupancy data with (epi)genomic and transcriptome data of primary GCs to define HNF4α target genes of in vitro and in vivo relevance. To investigate mechanistic roles of HNF4α and HNF4α targets, we performed cancer metabolic measurements, drug treatments and functional assays including murine xenograft experiments., Results: Gene expression analysis across 19 tumour types revealed HNF4α to be specifically upregulated in GCs. Unbiased pathway analysis revealed organic acid metabolism as the top HNF4α -regulated pathway, orthogonally supported by metabolomic analysis. Isocitrate dehydrogenase 1 ( IDH1 ) emerged as a convergent HNF4α direct target gene regulating GC metabolism. We show that wild-type IDH1 is essential for GC cell survival, and that certain GC cells can be targeted by IDH1 inhibitors., Conclusions: Our results highlight a role for HNF4α in sustaining GC oncogenic metabolism, through the regulation of IDH1 . Drugs targeting wild-type IDH1 may thus have clinical utility in GCs exhibiting HNF4α overexpression, expanding the role of IDH1 in cancer beyond IDH1/2 mutated malignancies., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. No commercial re-use. See rights and permissions. Published by BMJ.)

Background: Genetic susceptibility is associated with nasopharyngeal carcinoma (NPC). We previously identified rare variants potentially involved in familial NPC and common variants significantly associated with sporadic NPC., Methods: We conducted targeted gene sequencing of 20 genes [16 identified from the study of multiplex families, three identified from a pooled analysis of NPC genome-wide association study (GWAS), and one identified from both studies] among 819 NPC cases and 938 controls from two case-control studies in Taiwan (independent from previous studies). A targeted, multiplex PCR primer panel was designed using the custom Ion AmpliSeq Designer v4.2 targeting the regions of the selected genes. Gene-based and single-variant tests were conducted., Results: We found that NPC was associated with combined common and rare variants in CDKN2A/2B ( P = 1.3 × 10 -4 ), BRD2 ( P = 1.6 × 10 -3 ), TNFRSF19 ( P = 4.0 × 10 -3 ), and CLPTM1L/TERT ( P = 5.4 × 10 -3 ). Such associations were likely driven by common variants within these genes, based on gene-based analyses evaluating common variants and rare variants separately (e.g., for common variants of CDKN2A/2B, P = 4.6 × 10 -4 ; for rare variants, P = 0.04). We also observed a suggestive association with rare variants in HNRNPU ( P = 3.8 × 10 -3 ) for NPC risk. In addition, we validated four previously reported NPC risk-associated SNPs., Conclusions: Our findings confirm previously reported associated variants and suggest that some common variants in genes previously linked to familial NPC are associated with the development of sporadic NPC., Impact: NPC-associated genes, including CLPTM1L/TERT, BRD2 , and HNRNPU , suggest a role for telomere length maintenance in NPC etiology., (©2019 American Association for Cancer Research.)

Alpinia oxyphylla is a traditional Chinese medicine widely used for treating diarrhea, ulceration, and enuresis. Moreover, A. oxyphylla is effective for cognitive function improvement and nerve regeneration. Multiple sclerosis (MS) is a chronic neuronal inflammatory autoimmune disease that commonly affects young adults in high-latitude regions. The aim of this study was to evaluate the beneficial effects of A. oxyphylla in an experimental autoimmune encephalomyelitis (EAE) mouse model, which is an extensively used model for human MS. The ethanolic extract of A. oxyphylla fruit (AO-1) was orally administered to EAE mice. Our results showed AO-1 significantly reduced EAE symptoms. Histopathological analysis showed AO-1 reduced demyelination, inflammation, gliosis, and axonal swelling in the spinal cord. Furthermore, immunohistochemistry and quantitative polymerase chain reaction (qPCR) studies revealed that the infiltration of CD4 + , CD8 + T cells, and CD11b + monocytes into the spinal cord decreased in the AO-1-treated group. Mechanistically, the Th1 transcription factor T-bet, Th17 transcription factor retinoic acid receptor-related orphan receptor γ (ROR γ t), and inflammatory cytokines interferon (IFN)- γ and interleukin (IL)-17 were reduced in the spinal cords of mice treated with AO-1. The expression levels of T-bet and ROR γ t were also lowered in the spleens of those mice. Further in vitro study showed AO-1 inhibited production of IFN- γ , IL-2, and tumor necrosis factor- α from MOG 35-55 -peptide-stimulated splenocytes. One component isolated from AO-1, yakuchinone A, inhibited IL-17 production in vitro and reduced EAE symptoms in the mice. Collectively, our results indicate that AO-1 ameliorated the severity of EAE in mice and may involve the regulation of Th1/Th17 response. A. oxyphylla warrants further investigation, particularly regarding its clinical benefits for MS.

Background: Utilization of alternative transcription start sites through alterations in epigenetic promoter regions causes reduced expression of immunogenic N-terminal peptides, which may facilitate immune evasion in early gastric cancer. We hypothesized that tumors with high alternate promoter utilization would be resistant to immune checkpoint inhibition in metastatic gastric cancer., Patients and Methods: Two cohorts of patients with metastatic gastric cancer treated with immunotherapy were analyzed. The first cohort (N = 24) included patients treated with either nivolumab or pembrolizumab. Alternate promoter utilization was measured using the NanoString® (NanoString Technologies, Seattle, WA, USA) platform on archival tissue samples. The second cohort was a phase II clinical trial of patients uniformly treated with pembrolizumab (N = 37). Fresh tumor biopsies were obtained, and transcriptomic analysis was carried out on RNAseq data. Alternate promoter utilization was correlated to T-cell cytolytic activity, objective response rate and survival., Results: In the first cohort 8 of 24 (33%) tumors were identified to have high alternate promoter utilization (APhigh), and this was used to define the APhigh tertile of the second cohort (13 APhigh of 37). APhigh tumors exhibited decreased markers of T-cell cytolytic activity and lower response rates (8% versus 42%, P = 0.03). Median progression-free survival was lower in the APhigh group (55 versus 180 days, P = 0.0076). In multivariate analysis, alternative promoter utilization was an independent predictor of immunotherapy survival [hazard ratio 0.29, 95% confidence interval 0.099-0.85, P = 0.024). Analyzing tumoral evolution through paired pre-treatment and post-treatment biopsies, we observed consistent shifts in alternative promoter utilization rate associated with clinical response., Conclusion: A substantial proportion of metastatic gastric cancers utilize alternate promoters as a mechanism of immune evasion, and these tumors may be resistant to anti-PD1 immune checkpoint inhibition. Alternate promoter utilization is thus a potential mechanism of resistance to immune checkpoint inhibition, and a novel predictive biomarker for immunotherapy., Trial Registration: ClinicalTrials.gov Identifier: NCT#02589496., (© The Author(s) 2019. Published by Oxford University Press on behalf of the European Society for Medical Oncology.)

Purpose: The purpose of this study was to investigate the IOP-lowering effects of the ITRI-E-212, a new Rho-associated protein kinase (ROCK) inhibitor. ITRI-E-212 improved fluid outflow through the trabecular meshwork and reduced IOP with transient and mild conjunctival hyperemia. ITRI-E-212 can potentially be developed into new antiglaucoma agents., Methods: ITRI-E-212 was selected from more than 200 amino-isoquinoline structures because of its adequate solubility and drug-loading percentage in eye drops. ITRI-E-212 has less than 50% inhibitory concentration (IC50) against ROCK2. The in vitro kinase inhibition was evaluated using the ADP-Glo kinase assay. A comprehensive analysis of the kinase inhibitor selectivity of ITRI-E-212 was performed using the KINOMEscan methodology. The IOP-lowering effect and tolerability of ITRI-E-212 were investigated in normotensive and ocular hypertensive rabbits. The pharmacokinetics study was performed in vivo in the aqueous humor (AH), and hyperemia was assessed., Results: ITRI-E-212 showed high in vitro inhibitory activity against ROCK2 and high specificity against AGC kinases. The mean IOP-lowering effect of ITRI-E-212 in normotensive and ocular hypertensive models was 24.9% and 28.6%, respectively; 1% ITRI-E-212 produced notable reductions in IOP that were sustained for at least 6 hours after each dose once per day. Only transient, mild hyperemia was observed. The compound extracted from the AH reached 78.4% ROCK2 kinase inhibition at 1 hour after dose administration and was sustained for 4 hours., Conclusions: ITRI-E-212 is a novel and highly specific ROCK2 inhibitor with the ability to lower IOP in animal models. It has favorable pharmacokinetic and ocular tolerability profiles with only minimal conjunctival hyperemia.

Wu Ji Bai Feng Pill (WJBFP) is a traditional Chinese medicine (TCM) complex formula, which has been widely used in the treatment of various gynecological disorders. However, the quality control of multiple components in WJBFP is challengeable by using the methods applicable to analysis of several phytochemicals in single herbs or simple herbal preparations. The purpose of this study is to establish an ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-MS/MS) method for the quantitative determination of 20 bioactive compounds in WJBFP. The modified chromatographic conditions were achieved on an Agilent Poroshell 120 EC-C 18 column with a gradient elution consisted of 0.1% formic acid in acetonitrile and 0.1% aqueous formic acid (v/v). All analytes were determined using a triple quadrupole mass spectrometry in positive or negative ionization modes with multiple reaction monitoring (MRM) mode. An UHPLC-MS/MS method was optimized and validated for linearity, limits of detection and quantification, precision, repeatability, stability and recovery. The proposed method was applied for the analysis of 20 compounds in 19 batches of commercial WJBFP products. principal component analysis and hierarchical cluster analysis were applied to evaluate intrinsic quality and to identify chemical markers most responsible for quality evaluation. In conclusion, the established method offered speedy and sensitive determination for 20 compounds and is helpful for chemical standardization of commercial WJBFP products., (Copyright © 2018. Published by Elsevier Taiwan LLC.)