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Chenxi Ouyang,1,2 Changjiang Meng,3 Fei Li,3 Shanshan Nie,3 Liying Gong,4 Yu Cao,5 Hong Yuan,3,5 Zeying Feng6 1School of Pharmacology, University of South China, Hengyang, Hunan, People’s Republic of China; 2The Affiliated Nanhua Hospital, Department of Pharmacy, Hengyang Medical School University of South China, Hengyang, Hunan, People’s Republic of China; 3Center of Clinical Pharmacology, The Third Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China; 4Department of Critical Care Medicine, The Third Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China; 5Department of Cardiology, The Third Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China; 6Clinical Trial Institution Office, Liuzhou Hospital of Guangzhou Women and Children’s Medical Center, Liuzhou, Guangxi, People’s Republic of ChinaCorrespondence: Zeying Feng, Clinical Trial Institution Office, Liuzhou Hospital of Guangzhou Women and Children’s Medical Center, No. 50 Boyuan Avenue, Liuzhou, Guangxi, 545000, People’s Republic of China, Email fengzeying@foxmail.comPurpose: Mechanistic studies showed that morphine may impair the antiplatelet effect of P2Y12 inhibitors. However, Several clinical studies with cardiovascular events as an outcome are contradictory, and the broader impact of this drug interaction on additional organ systems remains uncertain. With multisource data, this study sought to determine the effects of morphine interaction with P2Y12 inhibitors on major adverse outcomes comprehensively, and identify the warning indicators.Patients and Methods: Interaction signals were sought in 187,919 safety reports from the FDA Adverse Event Reporting System (FAERS) database, utilizing reporting odds ratios (repOR). In a cohort of 5240 acute coronary syndrome patients, the analyses were validated, and the biological effects of warning indicators were further studied with Mendelian randomization and mediation analysis.Results: Potential risk of renal system adverse events in patients cotreated with morphine is significantly higher in FAERS (repOR 4.83, 95% CI 4.42– 5.28, false discovery rate adjusted-P =3.55*10− 209). The analysis of in-house patient cohorts validated these results with an increased risk of acute kidney injury (adjusted OR: 1.65; 95% CI: 1.20 to 2.26), and we also found a risk of myocardial infarction in patients treated with morphine (adjusted OR: 1.55; 95% CI: 1.14 to 2.11). The Morphine group exhibited diminished Plateletcrit (PCT) levels post-surgery and lower PCT levels were associated with an increased risk of AKI.Conclusion: The administration of morphine in patients treated with P2Y12 receptor inhibitors should be carefully evaluated. PCT may serve as a potential warning indicator for morphine-related renal injury.Keywords: P2Y12 receptor inhibitors, morphine, kidney injury, Mendelian randomization

Huanxin Liu,1 Cuiqiao Meng,1 Zhongli Wang,1 Chenghao Liu,1 Chunhong Yu,1 Zhong Li2 1Health Examination Center, Hebei General Hospital, Shijiazhuang, Hebei, 050051, People’s Republic of China; 2Department of General Surgery, Shijiazhuang People’s Hospital, Shijiazhuang, Hebei, 050011, People’s Republic of ChinaCorrespondence: Huanxin Liu, Health Examination Center, Hebei General Hospital, Shijiazhuang, Hebei, 050051, People’s Republic of China, Tel +86 13513317389, Email m13513317389@163.comObjective: Insulin resistance (IR) has a bearing on blood pressure (BP). Nevertheless, research on the relationships between surrogates for IR and BP is limited. In this study, we investigate the associations of these novel IR indices with BP in new-onset hypertension (HTN) and elevated BP individuals.Methods: An overall sample of 55,381 adult subjects was included in Hebei General Hospital. BP and other clinical indicators were measured. Triglyceride glucose (TyG) index, TyG-waist circumference (TyG-WC), TyG-body mass index (TyG-BMI), TyG-waist to height ratio (TyG-WHtR), triglyceride-to-high-density lipoprotein-cholesterol ratio (TG/HDL-C) and metabolic score for IR (METS-IR) were collected as dependable surrogates for IR. Examinees were categorized into four groups based on BP levels. Those involved were classified as quartiles according to the levels of six surrogate IR indices. Logistic regression analysis was adopted to evaluate the impact of substitute IR indicators on BP. The receiver operating characteristic curve (ROC) analysis was performed to explore the predictive ability of the parameters on BP.Results: The incidence of elevated BP, stage 1 HTN and stage 2 HTN was 7.86%, 24.05% and 23.76%, respectively. As the levels of six substitute IR indices rose, so did the BP. In the logistic regression analysis, after full adjustment, all alternative IR indicators were independently related to both stage 1 HTN and stage 2 HTN. Except for TG/HDL-C, other substitute IR indices were strongly associated with elevated BP. ROC curves analysis suggested TyG-WC and TyG-WHtR outperformed other indicators with higher odd ratios and area under the curve (AUC) in all the participants.Conclusion: Increased substitute IR indices were significantly associated with elevated BP in new-onset HTN and elevated BP individuals. TyG-WC and TyG-WHtR could better predict elevated BP, stage 1 HTN and stage 2 HTN.Keywords: insulin resistance, hypertension, elevated blood pressure, triglyceride glucose index, metabolic score for insulin resistance

Abstract Background Age is the strongest risk factor for the development of Alzheimer’s disease (AD). Besides the pathological hallmarks of β-amyloid (Aβ) plaques and neurofibrillary tangles, emerging evidence demonstrates a critical role of microglia and neuroinflammation in AD pathogenesis. Oleoylethanolamide (OEA) is an endogenous lipid amide that has been shown to promote lifespan and healthspan in C. elegans through regulation of lysosome-to-nucleus signaling and cellular metabolism. The goal of our study was to determine the role of OEA in the mediation of microglial activity and AD pathology using its stable analog, KDS-5104. Methods We used primary microglial cultures and genetic and pharmacological approaches to examine the signaling mechanisms and functional roles of OEA in mediating Aβ phagocytosis and clearance, lipid metabolism and inflammasome formation. Further, we tested the effect of OEA in vivo in acute LPS-induced neuroinflammation and by chronic treatment of 5xFAD mice. Results We found that OEA activates PPARα signaling and its downstream cell-surface receptor CD36 activity. In addition, OEA promotes TFEB lysosomal function in a PPARα-dependent but mTORC1-independent manner, the combination of which leads to enhanced microglial Aβ uptake and clearance. These are associated with the suppression of LPS-induced lipid droplet accumulation and inflammasome activation. Chronic treatment of 5xFAD mice with KDS-5104 restored dysregulated lipid profiles, reduced reactive gliosis and Aβ pathology and rescued cognitive impairments. Conclusion Together, our study provides support that augmenting OEA-mediated lipid signaling may offer therapeutic benefit against aging and AD through modulating lipid metabolism and microglia phagocytosis and clearance.

Lysosomes are active sites to integrate cellular metabolism and signal transduction. A collection of proteins associated with the lysosome mediate these metabolic and signaling functions. Both lysosomal metabolism and lysosomal signaling have been linked to longevity regulation; however, how lysosomes adjust their protein composition to accommodate this regulation remains unclear. Using deep proteomic profiling, we systemically profiled lysosome-associated proteins linked with four different longevity mechanisms. We discovered the lysosomal recruitment of AMP-activated protein kinase and nucleoporin proteins and their requirements for longevity in response to increased lysosomal lipolysis. Through comparative proteomic analyses of lysosomes from different tissues and labeled with different markers, we further elucidated lysosomal heterogeneity across tissues as well as the increased enrichment of the Ragulator complex on Cystinosin-positive lysosomes. Together, this work uncovers lysosomal proteome heterogeneity across multiple scales and provides resources for understanding the contribution of lysosomal protein dynamics to signal transduction, organelle crosstalk, and organism longevity.

Huanxin Liu,1 Yuling Xing,2 Qian Nie,1 Zhong Li,3 Cuiqiao Meng,1 Huijuan Ma2,4 1Health Examination Center, Hebei General Hospital, Shijiazhuang, Hebei, 050051, People’s Republic of China; 2Department of Endocrinology, Hebei General Hospital, Shijiazhuang, Hebei, 050051, People’s Republic of China; 3Department of General Surgery, Shijiazhuang People’s Hospital, Shijiazhuang, 050011, People’s Republic of China; 4Key Laboratory of Metabolic Disease in Hebei Province, Hebei General Hospital, Shijiazhuang, Hebei, 050051, People’s Republic of ChinaCorrespondence: Huijuan Ma, Department of Endocrinology, Hebei General Hospital, Shijiazhuang, 050051, People’s Republic of China, Tel +86 18032838686, Email huijuanma76@163.comObjective: Thyroid hormones (THs) exert instrumental effects in regulating lipids metabolism. Whereas, research investigating the relationship between sensitivity indices to THs and metabolic dysfunction-associated fatty liver disease (MAFLD) have contradicted this. This study was designed to approach the correlation between sensitivity indices to THs and MAFLD in euthyroid subjects.Methods: An overall sample of 6356 euthyroid participants were enrolled in a Chinese hospital. Free triiodothyronine to free thyroxine ratio (FT3/FT4), thyrotropin triiodothyronine resistance index (TT3RI), thyrotropin thyroxine resistance index (TT4RI), thyroid stimulating hormone index (TSHI) and thyroid feedback quantile-based indices (TFQIFT3 and TFQIFT4) were collected as sensitivity indicators to THs. Participants were split into two groups based on whether they suffered with MAFLD or not. And participants were categorized into quartiles based on sensitivity indicators to THs. The effects of sensitivity indices to THs on MAFLD were analyzed using regression analysis. Bootstrap was performed to assess the mediation effect of triglyceride glucose (TyG) index on the relationship between sensitivity parameters to THs and MAFLD.Results: The incidence of MAFLD in euthyroid subjects was 34.47%. As FT3/FT4, TT3RI and TFQIFT3 levels rose, so did the MAFLD prevalence. After adjustment for confounders, logistic regression analyses indicated that the high-level FT3/FT4 and TFQIFT3 still remained risk factors for MAFLD. The relevance of FT3/FT4 and MAFLD was stronger among those whose age ≤ 40 years and had non-visceral obesity. And the interrelation between TFQIFT3 and MAFLD was stronger in subjects whose age ≤ 40 years. Mediation analyses suggested that TyG index had a noteworthy indirect impact on the relationship between FT3/FT4, TFQIFT3 and MAFLD.Conclusion: Increased FT3/FT4 and TFQIFT3 were significantly related to MAFLD prevalence in populations with normal thyroid function. TyG index partly mediated the relevance between FT3/FT4, TFQIFT3 and MAFLD.Keywords: thyroid hormone sensitivity, metabolic dysfunction-associated fatty liver disease, triglyceride glucose index, mediation effect

Cuiqiao Meng,1 Yuling Xing,2,3 Lijing Huo,4 Huijuan Ma2,5,6 1Health Examination Center, Hebei General Hospital, Shijiazhuang, People’s Republic of China; 2Department of Endocrinology, Hebei General Hospital, Shijiazhuang, People’s Republic of China; 3Graduate School of Hebei Medical University, Shijiazhuang, People’s Republic of China; 4Department of Clinical Laboratory, Hebei General Hospital, Shijiazhuang, People’s Republic of China; 5Hebei Key Laboratory of Metabolic Diseases, Hebei General Hospital, Shijiazhuang, Hebei, People’s Republic of China; 6Department of Internal Medicine, Hebei Medical University, Shijiazhuang, Hebei, People’s Republic of ChinaCorrespondence: Huijuan Ma, Department of Endocrinology, Hebei General Hospital, Shijiazhuang, 050051, People’s Republic of China, Tel +86 18032838686, Email huijuanma76@163.comPurpose: To investigate the association between diabetic retinopathy (DR), DR intensity, and estimated glucose disposal rate (eGDR) in individuals with type 2 diabetes mellitus (T2DM).Patients and Methods: This study comprised 1762 T2DM patients who were admitted between January and December, 2021. Overall, the DR was identified in 430 patients. Based on the eGDR, the participants were divided into four study groups. One-way analysis of variance was used to compare the groups. The correlations between eGDR and DR risk, eGDR, and DR severity were analyzed using regression analysis. Furthermore, these relationships were analyzed in different sex groups.Results: Patients with T2DM had a 19.75% (348/1762) DR detection rate, whereas those with DR had a 22.41% (78/348) proliferative DR detection rate. The DR group had substantially reduced levels of eGDR compared with the non-DR group. Multivariate logistic regression analysis demonstrated that reduced eGDR was an independent risk factor for DR, after adjusting for confounding variables. eGDR correlated significantly with proliferative DR in women but not in men.Conclusion: In Chinese individuals with T2DM, lower eGDR was independently associated with a higher risk of DR.Keywords: estimated glucose disposal rate, type 2 diabetes mellitus, diabetic retinopathy

Microglia are the major cell type expressing complement C3a receptor (C3aR) in the brain. Using a knockin mouse line in which a Td-tomato reporter is incorporated into the endogenous C3ar1 locus, we identified 2 major subpopulations of microglia with differential C3aR expression. Expressing the Td-tomato reporter on the APPNL-G-F–knockin (APP-KI) background revealed a significant shift of microglia to a high-C3aR-expressing subpopulation and they were enriched around amyloid β (Aβ) plaques. Transcriptomic analysis of C3aR-positive microglia documented dysfunctional metabolic signatures, including upregulation of hypoxia-inducible factor 1 (HIF-1) signaling and abnormal lipid metabolism in APP-KI mice compared with wild-type controls. Using primary microglial cultures, we found that C3ar1-null microglia had lower HIF-1α expression and were resistant to hypoxia mimetic–induced metabolic changes and lipid droplet accumulation. These were associated with improved receptor recycling and Aβ phagocytosis. Crossing C3ar1-knockout mice with the APP-KI mice showed that C3aR ablation rescued the dysregulated lipid profiles and improved microglial phagocytic and clustering abilities. These were associated with ameliorated Aβ pathology and restored synaptic and cognitive function. Our studies identify a heightened C3aR/HIF-1α signaling axis that influences microglial metabolic and lipid homeostasis in Alzheimer disease, suggesting that targeting this pathway may offer therapeutic benefit.

Abstract This study introduces a deep learning approach to predicting demographic features from meibography images. A total of 689 meibography images with corresponding subject demographic data were used to develop a deep learning model for predicting gland morphology and demographics from images. The model achieved on average 77%, 76%, and 86% accuracies for predicting Meibomian gland morphological features, subject age, and ethnicity, respectively. The model was further analyzed to identify the most highly weighted gland morphological features used by the algorithm to predict demographic characteristics. The two most important gland morphological features for predicting age were the percent area of gland atrophy and the percentage of ghost glands. The two most important morphological features for predicting ethnicity were gland density and the percentage of ghost glands. The approach offers an alternative to traditional associative modeling to identify relationships between Meibomian gland morphological features and subject demographic characteristics. This deep learning methodology can currently predict demographic features from de-identified meibography images with better than 75% accuracy, a number which is highly likely to improve in future models using larger training datasets, which has significant implications for patient privacy in biomedical imaging.

Aims: There is an increasing concern of osteoporotic fractures in the ageing population. Low-magnitude high-frequency vibration (LMHFV) was shown to significantly enhance osteoporotic fracture healing through alteration of osteocyte lacuno-canalicular network (LCN). Dentin matrix protein 1 (DMP1) in osteocytes is known to be responsible for maintaining the LCN and mineralization. This study aimed to investigate the role of osteocyte-specific DMP1 during osteoporotic fracture healing augmented by LMHFV. Methods: A metaphyseal fracture was created in the distal femur of ovariectomy-induced osteoporotic Sprague Dawley rats. Rats were randomized to five different groups: 1) DMP1 knockdown (KD), 2) DMP1 KD + vibration (VT), 3) Scramble + VT, 4) VT, and 5) control (CT), where KD was performed by injection of short hairpin RNA (shRNA) into marrow cavity; vibration treatment was conducted at 35 Hz, 0.3 g; 20 minutes/day, five days/week). Assessments included radiography, micro-CT, dynamic histomorphometry and immunohistochemistry on DMP1, sclerostin, E11, and fibroblast growth factor 23 (FGF23). In vitro, murine long bone osteocyte-Y4 (MLO-Y4) osteocyte-like cells were randomized as in vivo groupings. DMP1 KD was performed by transfecting cells with shRNA plasmid. Assessments included immunocytochemistry on osteocyte-specific markers as above, and mineralized nodule staining. Results: Healing capacities in DMP1 KD groups were impaired. Results showed that DMP1 KD significantly abolished vibration-enhanced fracture healing at week 6. DMP1 KD significantly altered the expression of osteocyte-specific markers. The lower mineralization rate in DMP1 KD groups indicated that DMP1 knockdown was associated with poor fracture healing process. Conclusion: The blockage of DMP1 would impair healing outcomes and negate LMHFV-induced enhancement on fracture healing. These findings reveal the importance of DMP1 in response to the mechanical signal during osteoporotic fracture healing. Cite this article: Bone Joint Res 2022;11(7):465–476.

Jingjing Liu,1– 3 Guoqing Fan,1– 3 Ningning Tao,4 Feifei Feng,5 Chao Meng,1– 3 Tieying Sun1,2 1Department of Respiratory Medicine and Critical Care, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 2Graduate School of Peking Union Medical College, Beijing, People’s Republic of China; 3The MOH Key Laboratory of Geriatrics, Beijing Hospital, National Center of Gerontology, Beijing, People’s Republic of China; 4Department of Respiratory & Critical Care Medicine, Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, Shandong, People’s Republic of China; 5Department of Respiratory & Critical Care Medicine, The Second Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong, People’s Republic of ChinaCorrespondence: Tieying Sun, Department of Respiratory Medicine and Critical Care, Beijing Hospital, National Center of Gerontology, Institute of Geriatric Medicine, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China, Tel +15153169108, Email suntieying@126.comPurpose: Idiopathic pulmonary fibrosis is a chronic and irreversible fibrotic interstitial pneumonia of unknown etiology and therapeutic strategies are limited. Emerging evidence suggests that the continuous activation of the central nucleotide-binding oligomerization-, leucine-rich repeat-, and pyrin domain-containing protein 3 (NLRP3) inflammasome is involved in the pathogenesis of pulmonary fibrosis. Ginsenoside Rb1 (G-Rb1) is the most abundant component in the traditional Chinese herb ginseng and has anti-inflammatory and anti-fibrotic activities. The purpose of this study was to explore whether G-Rb1 exerts anti-inflammatory and anti-fibrotic activities in vivo and in vitro by suppressing the activation of the NLRP3 inflammasome and NF-κB pathway.Methods: Forty-eight male C57BL/6 mice were randomly divided into four groups (n=12/group) as follows: control, bleomycin (BLM), BLM/G-Rb1, and G-Rb1. A pulmonary fibrosis model was developed via an intratracheal injection of BLM. Six mice from each group were euthanized on days 3 and 21. The degree of pulmonary fibrosis was examined by histological evaluation and assessing α-smooth muscle actin levels. THP-1 cells were differentiated into macrophages, and stimulated by lipopolysaccharide and adenosine triphosphate. Activation of the NLRP3 inflammasome and NF-κB pathway was determined by Western blotting. Interleukin-1 beta and interleukin-18 levels were measured by ELISA. MRC-5 cells were cultured in the conditioned medium of the treated macrophages, after which markers of myofibroblasts were determined by Western blotting.Results: G-Rb1 ameliorated BLM-induced pulmonary inflammation and fibrosis in mice, and suppressed NLRP3 inflammasome activation and the NF-κB pathway in lung tissues. Moreover, interleukin-1 beta secreted after NLRP3 inflammasome activation in macrophages promoted fibroblast differentiation. G-Rb1 inhibited lipopolysaccharide- and adenosine triphosphate-induced NLRP3 inflammasome activation in macrophages and disturbed the crosstalk between macrophages and fibroblasts.Conclusion: G-Rb1 ameliorates BLM-induced pulmonary inflammation and fibrosis by suppressing NLRP3 inflammasome activation and the NF-κB pathway. Hence, G-Rb1 is a potential novel therapeutic drug for idiopathic pulmonary fibrosis.Keywords: G-Rb1, pulmonary fibrosis, NLRP3 inflammasome, macrophages

Microglia are the major cell type expressing complement C3a receptor (C3aR) in the brain. Using a knockin mouse line in which a Td-tomato reporter is incorporated into the endogenous C3ar1 locus, we identified 2 major subpopulations of microglia with differential C3aR expression. Expressing the Td-tomato reporter on the [APP.sup.NL-G-F]-knockin (APP-KI) background revealed a significant shift of microglia to a high-C3aR-expressing subpopulation and they were enriched around amyloid [beta] (A[beta]) plaques. Transcriptomic analysis of C3aR-positive microglia documented dysfunctional metabolic signatures, including upregulation of hypoxia-inducible factor 1 (HIF-1) signaling and abnormal lipid metabolism in APP-KI mice compared with wild-type controls. Using primary microglial cultures, we found that C3ar1-null microglia had lower HIF-1[alpha] expression and were resistant to hypoxia mimetic-induced metabolic changes and lipid droplet accumulation. These were associated with improved receptor recycling and A[beta] phagocytosis. Crossing C3ar1-knockout mice with the APP-KI mice showed that C3aR ablation rescued the dysregulated lipid profiles and improved microglial phagocytic and clustering abilities. These were associated with ameliorated A[beta] pathology and restored synaptic and cognitive function. Our studies identify a heightened C3aR/HIF-1[alpha] signaling axis that influences microglial metabolic and lipid homeostasis in Alzheimer disease, suggesting that targeting this pathway may offer therapeutic benefit., Introduction Alzheimer disease (AD), the leading cause of dementia in the elderly, is pathologically characterized by the accumulation of amyloid [beta] (A[beta]) plaques and intracellular tau tangles and accompanying neuroinflammation [...]

Xiaoling Liu,1,2,&ast; Junhua Mai,1,&ast; Chaoyang Meng,1 Aldona J Spiegel,3 Wei Wei,2 Haifa Shen1 1Department of Nanomedicine, Houston Methodist Academic Institute, Houston, TX, 77030, USA; 2Department of Breast and Thyroid Surgery, Peking University Shenzhen Hospital, Shenzhen, Guangdong, 518036, People’s Republic of China; 3Institute for Reconstructive Surgery, Houston Methodist Hospital, Weill Cornell Medicine, Houston, TX, USA&ast;These authors contributed equally to this workCorrespondence: Haifa ShenDepartment of Nanomedicine, Houston Methodist Academic Institute, Houston, TX, 77030, USATel +1 713-441-7321Email HaifaShen@gmail.comBackground: Abdominal flaps are routinely performed in clinic after primary mastectomy of breast cancer. However, cancer patients can still develop cancer recurrence and metastasis after surgery. In this study, we evaluated the feasibility of concurrent abdominal flap reconstruction and vaccine inoculation in the tissue for prevention and treatment of HER2-positive breast cancer.Methods: A murine model of metastatic HER2-positive breast cancer was generated by inoculating HER2-expressing TUBO tumor cells into both the mammary gland fat pad and left ventricle. Mammary gland fat pad with primary tumor was resected by mastectomy, and superficial inferior epigastric (SIE) vessel-based abdominal flap was performed for abdominal reconstruction. During the surgery, mice also received a single intra-flap treatment of a microparticulate-based cancer vaccine. Popliteal (Pop) and inguinal (Ing) lymph nodes (LN) were collected at different time points after vaccination, and activation of dendritic cells and T lymphocytes was evaluated with flow cytometry. ELISpot was also performed to measure HER2-specific T cells in splenocytes. In addition, infiltration of CD3+ T cells in brain metastatic nodules was analyzed with immunohistochemistry.Results: Flow cytometry detected increased number of activated dendritic cells in lymph nodes in mice treated with cancer vaccine. ELISpot revealed abundant IFN-γ-expressing T cells in the spleen. Mice treated with abdominal flap-embedded cancer vaccine extended median survival by 9 days over the control group (p< 0.05).Conclusion: Abdominal flap-embedded cancer vaccine effectively stimulated systemic immune response and inhibited tumor progression in a murine model of HER2-positive breast cancer.Keywords: silicon microparticle, cancer vaccine, peddled flap-based reconstruction, breast cancer, HER2

Abstract The surface of the human eye is covered with a protective tear film that refreshes with each blink. Natural blinking occurs involuntarily, but one can also voluntarily blink or refrain from blinking. The maximum time one can refrain from blinking until the onset of discomfort is the maximum interblink period (MIBP). During the interblink period the tear film evaporates and thins from the ocular surface. Infrared thermography provides a non-invasive measure of the ocular surface temperature (OST). Due to evaporation, ocular surface cooling (OSC) generally occurs when the eyes are open and exposed to the environment. The purpose of our study was to investigate the effect of OSC rate on the MIBP, and to investigate the association of the MIBP with tear film characteristics in subjects who do and do not exhibit OSC. The MIBP was measured simultaneously with OST over time. Non-invasive tear breakup time, tear meniscus height, tear lipid layer thickness, and Schirmer I test strip wetted lengths were measured on a day prior to the thermography visit. Subjects were divided into cooling and non-cooling groups based on OSC rate, and demographic and tear film characteristics were tested for inter-group differences. A faster OSC rate was associated with an exponentially shorter duration of the MIBP overall and within the cooling group alone. Faster non-invasive tear breakup time was significantly associated with a shorter MIBP in both groups. These results suggest that tear film evaporation initiates a pathway that results in the onset of ocular discomfort and the stimulus to blinking. The presence of a subset of subjects with no or minimal OSC who nevertheless have a short MIBP indicates that evaporative cooling is not the only mechanism responsible for the onset of ocular discomfort.

Luoning Gou,1,* Ming Xiang,2,* Xiao Ran,3,* Fen Wang,1 Shujun Zhang,1 Shusheng Li,3 Kun Dong,1 Xi Chen,1 Yangxin Huang,4 Chengzhen Meng,2 Qian Fan,2 Yan Yang,1 Xuefeng Yu,1 Delin Ma,1 Ping Yin2 1Department of Endocrinology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 2Department of Epidemiology and Biostatistics, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 3Department of Emergency Medicine, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People’s Republic of China; 4College of Public Health, University of South Florida, Tampa, Florida, The United States*These authors contributed equally to this workCorrespondence: Ping Yin; Delin MaTongji Medical College, Huazhong University of Science and Technology, 13 Hongkong Road, Wuhan, 430030 Hubei, People’s Republic of ChinaTel + 86-27-83692832Fax +86-27-83692333Email pingyin2000@126.comDelin Ma Email maderine4@163.comPurpose: Recently, a cluster of pneumonia caused by SARS-CoV-2 were identified in Wuhan and spread throughout the world. More information about risk factors for mortality of critically ill patients infected with SARS-CoV-2 remain to be evaluated.Methods: We included adult patients confirmed with SARS-CoV-2 infection who were critically ill and admitted to the intensive care unit (ICU) of Tongji Hospital in Wuhan from Feb 4, 2020 to Feb 20, 2020. Data were collected and compared between patients who died and improved. Logistic regression was used to explore the risk factors for death of SARS-CoV-2-infected critically ill patients.Results: A total of 160 critically ill patients with SARS-CoV-2 infection were included, of which 146 patients with appeared outcomes were included into the final analysis. The random blood glucose, serum sodium and effective plasma osmolarity were higher in deceased patients, especially in patients with diabetes. There were 7 patients with diabetes with hyperosmolar status and all of them were deceased. Multivariable regression revealed that older age (odds ratio 4.28, 95% CI 1.01– 18.20; p = 0.049), higher C-reactive protein (odds ratio 1.01, 1.00– 1.03; p = 0.024), higher interleukin-6 (odds ratio 1.01, 1.00– 1.03; p = 0.0323), and d-dimer greater than 1 μg/mL (odds ratio 1.10, 1.01– 1.20; p = 0.032) at admission were associated with increased odds of death.Conclusion: In conclusion, hyperosmolarity needs more attention and may contribute to mortality in critically ill patients with COVID-19, especially in those with diabetes. Older age, inflammatory response, and thrombosis may be risk factors for death of critically ill patients with SARS-CoV-2 infection.Keywords: SARS-CoV-2, critically ill, hyperosmolarity, diabetes, risk factors, mortality

PROTACs have emerged as promising therapeutic agents but their cellular uptake is often inefficient. Here, the authors show that reversible covalent warhead chemistry improves PROTAC intracellular accumulation and target engagement, and develop a dual inhibitor/degrader of Bruton’s tyrosine kinase

Mitochondrial permeability transition pore (MPTP)-dependent necrosis contributes to numerous pathologies in the heart, brain, and skeletal muscle. The MPTP is a non-selective pore in the inner mitochondrial membrane that is triggered by high levels of matrix Ca2+, and sustained opening leads to mitochondrial dysfunction. Although the MPTP is defined by an increase in inner mitochondrial membrane permeability, the expression of pro-apoptotic Bcl-2 family members, Bax and Bak localization to the outer mitochondrial membrane is required for MPTP-dependent mitochondrial dysfunction and subsequent necrotic cell death. Contrary to the role of Bax and Bak in apoptosis, which is dependent on their oligomerization, MPTP-dependent necrosis does not require oligomerization as monomeric/inactive forms of Bax and Bak can facilitate mitochondrial dysfunction. However, the relationship between Bax and Bak activation/oligomerization and MPTP sensitization remains to be explored. Here, we use a combination of in vitro and ex vivo approaches to determine the role of the anti-apoptotic Bcl-2 family members, which regulate Bax/Bak activity, in necrotic cell death and MPTP sensitivity. To study the role of each predominantly expressed anti-apoptotic Bcl-2 family member (i.e., Mcl-1, Bcl-2, and Bcl-xL) in MPTP regulation, we utilize various BH3 mimetics that specifically bind to and inhibit each. We determined that the inhibition of each anti-apoptotic Bcl-2 family member lowers mitochondrial calcium retention capacity and sensitizes MPTP opening. Furthermore, the inhibition of each Bcl-2 family member exacerbates both apoptotic and necrotic cell death in vitro in a Bax/Bak-dependent manner. Our findings suggests that mitochondrial Ca2+ retention capacity and MPTP sensitivity is influenced by Bax/Bak activation/oligomerization on the outer mitochondrial membrane, providing further evidence of the crosstalk between the apoptotic and necrotic cell death pathways.

Hypothesis: Biophysical property and water evaporation retardation through a lipid nanofilm can be altered by model tear protein and topical ophthalmic formulation. Experiment: Evaporation rate and dynamic surface pressure were measured using a sessile drop technique. Water evaporations from 5 individual protein solutions, their mixture, and 6 ophthalmic formulations were quantified. Biophysical property and evaporation through model lipid nanofilms spread on model electrolyte solutions, tear protein solutions, and ophthalmic formulations were assessed. Findings: Model lipid nanofilms spread on electrolyte solution reduced evaporative fluxes by 43–60%. Evaporative fluxes from individual protein solutions without lipids were 3–19% lower than from electrolytes solution. Evaporative fluxes through lipid nanofilms were decreased by the presence of albumin or lactoferrin in solutions but increased by lysozyme and mucin.Evaporative fluxes from ophthalmic formulations were 10–43% lower than from electrolyte solution. Evaporations through lipid nanofilms spread on formulations were higher than through lipids on electrolyte solution. Model lipid nanofilms spread on Diquas appeared more rigid than on electrolyte solution but showed softening when spread on Refresh Mega-3.Some proteins and ophthalmic formulations altered model lipid nanofilms evaporative barriers. Ophthalmic formulation induced changes in biophysical property of model lipid nanofilms in vitro may suggest possible tear film destabilization in vivo.

Olfaction is a key sensory modality with high diversity and olfactory defects has been associated with metabolic and neurodegenerative disorders. Here, the authors discovered that specific olfactory inputs actively regulate lipid metabolism in a dynamic and reversible manner.

Cui Meng,1–3* Weipeng Wei,1,2* Yuhe Wang,1,3 Kunqin Zhang,1,2 Ting Zhang,1,2 Yunyan Tang,1,2 Fushan Tang1,2 1Department of Clinical Pharmacy, School of Pharmacy, Zunyi Medical University, Zunyi 563006, People’s Republic of China; 2The Key Laboratory of Clinical Pharmacy in Zunyi City, Zunyi 563006, People’s Republic of China; 3Department of Pharmacy, The First Hospital Affiliated to Zunyi Medical University, Zunyi 563003, People’s Republic of China*These authors contributed equally to this workCorrespondence: Fushan TangSchool of Pharmacy, Zunyi Medical University, Zunyi 563006, People’s Republic of ChinaTel +8685128642337Email fstang@vip.163.comObjective: This study aimed to investigate the interaction between the ion-complementary self-assembling peptide RADA16-I and the hydrophobic drug mangiferin (MA), and the potential of the self-assembling peptide to be exploited as a drug carrier of MA.Methods: The RADA16-I–MA suspension was prepared by magnetic stirring, followed by fluorescence spectrophotometry, particle size determination, rheological properties analysis, and in vitro release assay to characterize the interaction between RADA16-I and MA. Then, the effects of in situ MA-loaded hydrogel on the proliferation of KYSE 30 and DLD-1 tumor cells and the toxic effect of the hydrogel on 293T renal epithelial cells were studied by the Cell Counting Kit 8 method.Results: The RADA16-I–MA suspension was formed in water under magnetic stirring; the in situ hydrogel was formed when the suspension was added to PBS. The particle size in the RADA16-I–MA suspension was around 300–600 nm with an average size of 492 nm. Within 24 h, the cumulative release of MA from the RADA16-I–MA hydrogel was about 80%. The release rate of MA from the hydrogel was dependent on the concentration of RADA16-I and the release can be fitted with a first-order kinetic equation. The results suggested that the self-assembling peptide can stabilize MA in water to form a relatively stable suspension; the results also indicated that controlled release of MA from the RADA16-I–MA in situ hydrogel formed from the RADA16-I–MA suspension can be achieved by adjusting the concentration of the peptide in suspension. The cell viability studies showed that the RADA16-I–MA in situ hydrogel not only can maintain or enhance the intrinsic proliferation inhibition effects of MA on tumor cells, but also can reduce the toxicity of MA to normal cells.Conclusion: The self-assembling peptide RADA16-I can be a potential candidate for constructing a delivery system of the hydrophobic drug MA.Keywords: RADA16-I, hydrophobic drug, delivery system, antitumor

Wenlong Wang*, Chaoyang Meng*, Qianhui Ouyang, Jing Xie, Xinying Li Division of Thyroid, General Surgery Department, Xiangya Hospital, Central South University, Changsha 410008, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xinying LiDivision of Thyroid, General Surgery Department, Xiangya Hospital, Central South University, No.87 Xiangya Road, Changsha 410008, People’s Republic of ChinaTel +86 07 318 975 3710Email lixinyingcn@126.comBackground: This study was designed to investigate the impact of serum magnesium (Mg) levels on hypocalcemia after thyroidectomy.Patients and methods: In total, 242 patients with differentiated thyroid cancer were retrospectively analyzed.Results: Multivariate regression analysis showed hypomagnesemia was an independent risk factor for hypocalcemia (P0.05). In patients with normal Po-Ca levels, hypomagnesemia increased the risk of symptoms related to hypocalcemia by 4.478 times (OR =5.478, 95% CI 1.724–17.403).Conclusion: Hypomagnesemia, or even a low serum Mg level within the normal range, can increase the risk of hypocalcemia. After excluding the potential effects of PTH on serum magnesium and calcium, serum Mg reduction is one of the most important factors that influences postoperative serum Ca reduction. What’s more, hypomagnesemia is closely linked with symptoms.Keywords: hypocalcemia, hypomagnesemia, thyroidectomy, magnesium, calcium