Fabrication of Stable Apigenin Nanosuspension with PEG 400 as Antisolvent for Enhancing the Solubility and Bioavailability.

The purpose of this paper is to prepare a stable apigenin nanosuspension with a drug concentration of 1.11 mg/mL through green and efficient antisolvent method. Compared with the traditional preparation process that may use toxic reagents, in this study, a green and effective strategy was applied for the preparation of stable apigenin nanosuspension by using an antisolvent method with PEG 400 as antisolvent to improve the solubility and bioavailability. It was found that the particle size of apigenin nanosuspension was about 280 nm, and the solubility and dissolution of the nanosuspension were 33 and 3 times higher than that of the apigenin, respectively. Pharmacokinetic study showed that the Cmax and AUC 0-8 h values of the nanosuspension in fasting rats achieved about 6- and 2.5-fold enhancement than that of the apigenin, respectively. Stability test showed that the apigenin nanosuspension could be stored stably for 12 months at 25℃. Taken together, the antisolvent method with PEG 400 was proven to be a green and effective method to prepare the stable nanosuspension of poorly soluble drugs.

Soluble E-cadherin is associated with oxidative stress in patients with chronic HBV infection.

Mounting evidence indicates that serum soluble E-cadherin (sE-cadherin) serves as an important player in various physiological and pathological processes. However, the crosstalk between serum sE-cadherin and oxidative stress in chronic hepatitis B (CHB) remains to be illustrated. The main purpose of this study is to explore the molecular mechanisms underlying the function of sE-cadherin in CHB virus infection. Levels of serum sE-cadherin, total antioxidant capacity (TAC), glutathione (GSH), superoxide dismutase (SOD), total oxidant activity (TOA), NADPH oxidase 2 (NOX2), and malondialdehyde (MDA), from 51 patients with hepatitis B envelope antigen (HBeAg)-negative CHB, 54 patients with HBeAg-positive CHB, and 109 healthy individuals were detected by enzyme-linked immunosorbent assay. In our study, patients with CHB showed significantly higher serum sE-cadherin levels than healthy individuals (P < .01). Furthermore, we also found that the serum sE-cadherin levels were significantly negatively correlated with TAC, antioxidant enzymes (GSH and SOD) in patients with CHB, and that serum sE-cadherin concentrations were significantly positively correlated with liver enzyme markers (alanine transaminase and aspartate aminotransferase) and oxidative markers (TOA, NOX2, and MDA) in patients with CHB. Therefore, serum sE-cadherin may act as a new candidate biomarker for reflecting inflammation and oxidative stress status in the development and progression of hepatitis B virus infection.

CircIL4R facilitates the tumorigenesis and inhibits ferroptosis in hepatocellular carcinoma by regulating the miR-541-3p/GPX4 axis.

Ferroptosis is a specific iron-dependent cell death form that can induce the production of lipid peroxide, but the roles of circular RNAs (circRNAs) in ferroptosis are completely unaware. Circ-interleukin-4 receptor (circIL4R) was reported to express highly in hepatocellular carcinoma (HCC). This study focused on the function of circIL4R dysregulation in tumor progression and ferroptosis of HCC, as well as its molecular mechanism. The quantitative real-time polymerase chain reaction was implemented for measuring RNA expression. Cell proliferation and survival were evaluated using 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide. Apoptotic cells were detected via flow cytometry. The quantification of protein expression was executed through western blotting analysis. The target binding was assessed via the dual-luciferase reporter, RNA immunoprecipitation, and RNA pull-down assays. The experiment in vivo was performed using a xenograft model. CircIL4R was abnormally overexpressed in HCC tissues and cells. CircIL4R knockdown impeded oncogenesis and expedited ferroptosis of HCC cells. CircIL4R could directly sponge microRNA-541-3p (miR-541-3p) and miR-541-3p inhibition mitigated the effects of circIL4R knockdown on HCC cells. CircIL4R acted as a miR-541-3p sponge to regulate its target glutathione peroxidase 4 (GPX4). GPX4 upregulation relieved the miR-541-3p-induced tumor inhibition and ferroptosis aggravation. CircIL4R played an oncogenic role in HCC via the miR-541-3p/GPX4 axis in vivo. Our data suggested that circIL4R served for a tumor promoter and ferroptosis inhibitor in HCC by the miR-541-3p/GPX4 network.

The ratio of superoxide dismutase to standard deviation of erythrocyte distribution width as a predictor of systemic lupus erythematosus.

BACKGROUND: To explore the clinical value of the serum superoxide dismutase-to-standard deviation of erythrocyte distribution width ratio (SRSR) in systemic lupus erythematosus (SLE). METHODS: A total of 222 SLE patients from the Rheumatology and Immunology Department in the Second Affiliated Hospital of Chongqing Medical University from January 2017 to April 2019 were collected as the experimental group, and a total of 202 healthy physical examiners were extracted as the control group. Neutrophil-to-lymphocyte ratio (NLR), superoxide dismutase-to-standard deviation of erythrocyte distribution width ratio (SRSR), and platelet-to-lymphocyte ratio (PLR) were calculated from the collected data and then compared the level of the above three indexes between the two groups. In addition, we analyzed the association between SRSR and clinically relevant indicators. RESULTS: We found that the SRSR of SLE patients was significantly lower than healthy control group, by analyzing the receiver operating characteristic (ROC) curve; it revealed that the SRSR had higher specificity and sensitivity than either superoxide dismutase (SOD) or standard deviation of erythrocyte distribution width (RDW-SD) alone. The area under the curve (AUC) for SRSR was significantly larger than either SOD or RDW-SD alone, and the AUC for SRSR was also larger than NLR and PLR. And it was found that SRSR was independently correlated with SLE disease activity through multiple linear regression analysis. CONCLUSION: SRSR is a useful biomarker for the diagnosis of SLE, and it is of great significance in the clinical application.

Overexpression of circ_0001445 decelerates hepatocellular carcinoma progression by regulating miR-942-5p/ALX4 axis.

Circular RNAs (circRNAs) have been verified to have essential regulatory roles in diverse human cancers, including hepatocellular carcinoma (HCC). In this study, we aimed to explore the roles of circ_0001445 in HCC. Herein, circ_0001445 was decreased and miR-942-5p was increased in HCC tissues and cells. Circ_0001445 overexpression or miR-942-5p inhibition repressed cell cycle process, migration, invasion, epithelial-mesenchymal transition and glycolysis in HCC cells. Mechanistically, circ_0001445 could promote ALX4 expression through targeting miR-942-5p. Moreover, miR-942-5p overexpression reversed the inhibitory effect of circ_0001445 on HCC cell progression. The effect of miR-942-5p on HCC cell development was rescued following the elevation of ALX4. In addition, circ_0001445 overexpression restrained tumorigenesis in vivo. In conclusion, circ_0001445 played a negative role in HCC progression by modulating miR-942-5p/ALX4 axis, which might provide a novel target for HCC therapy.

The association between hypoxia inducible factor 1 subunit alpha gene rs2057482 polymorphism and cancer risk: a meta-analysis.

BACKGROUND: The rs2057482 polymorphism in the hypoxia inducible factor 1 subunit alpha (HIF1A) gene has been reported to be associated with a risk of several types of cancer, but this association has not yet been definitively confirmed. We performed this meta-analysis to determine whether rs2057482 is associated with overall cancer risk. METHODS: The PubMed, Embase, and Web of Science databases were searched for the potential studies about the association between the rs2057482 and cancer risk. The data of genotype frequencies in cases with cancer and controls were extracted from the selected studies. Odds ratios (ORs) and the corresponding 95% confidence intervals (CIs) were calculated to determine the strength of the associations. RESULTS: The meta-analysis showed an association between the rs2057482 polymorphism and overall cancer risk. However, a stratified analysis of ethnicity did not show any significant association between rs2057482 and cancer risk in the Asian population. CONCLUSIONS: The rs2057482 polymorphism was associated with decreased overall cancer risk, based on the currently available studies. However, this conclusion needs verification by further well-designed epidemiology studies that examine different cancer types and more subjects.

BeeKeeper 2.0: Confidential Blockchain-Enabled IoT System with Fully Homomorphic Computation.

Blockchain-enabled Internet of Things (IoT) systems have received extensive attention from academia and industry. Most previous constructions face the risk of leaking sensitive information since the servers can obtain plaintext data from the devices. To address this issue, in this paper, we propose a decentralized outsourcing computation (DOC) scheme, where the servers can perform fully homomorphic computations on encrypted data from the data owner according to the request of the data owner. In this process, the servers cannot obtain any plaintext data, and dishonest servers can be detected by the data owner. Then, we apply the DOC scheme in the IoT scenario to achieve a confidential blockchain-enabled IoT system, called BeeKeeper 2.0. To the best of our knowledge, this is the first work in which servers of a blockchain-enabled IoT system can perform any-degree homomorphic multiplications and any number of additions on encrypted data from devices according to the requests of the devices without obtaining any plaintext data of the devices. Finally, we provide a detailed performance evaluation for the BeeKeeper 2.0 system by deploying it on Hyperledger Fabric and using Hyperledger Caliper for performance testing. According to our tests, the time consumed between the request stage and recover stage is no more than 3.3 s, which theoretically satisfies the production needs.

Effects of lily/maize intercropping on rhizosphere microbial community and yield of Lilium davidii var. unicolor.

Continuous cropping of lily (Lilium davidii var. unicolor) or any other crop seriously affects yield and quality. In this study, we compared continuous cropping with lily/maize intercropping. We also examined the lily rhizosphere microbes community in both sole lily cropping and lily/maize intercropping systems, by the llumina Miseq platform. Here we refer to data of recent years field experimentation of a lily/maize intercrop system in different planting configurations in the Gaolan Ecological and Agricultural Research Station. Treatments included sole crops of lily and maize, an intercrop consisting of strips of four lily rows alternating with one maize rows. The land equivalent ratio (LER) of intercrops was 1.294. The results showed that compared to sole cropping, the yield of lily in the first year of planting increased when lily was intercropped with maize. The species annotation of the high-throughput sequencing experiment showed that there was no difference in the diversity of the lily rhizosphere soil microbes on phylum taxonomic level, but the relative abundance of some genus changed obviously. The relative abundance of harmful fungus Fusarium spp. and, Funneliformis spp., decreased, and the relative abundance of beneficial bacteria Sphingomonas spp. and, Nitrospira spp., increased. In addition, we found that Lecanicillium spp., appeared only in the intercropping lily rhizosphere soil and sole cropping maize rhizosphere soil. In conclusion, the findings indicated that lily/maize intercropping could change soil microenvironment, and affect the diversity and structure of microorganism community in lily rhizosphere, with further beneficial effect on the yield of lily.

MIStore: a Blockchain-Based Medical Insurance Storage System.

Currently, blockchain technology, which is decentralized and may provide tamper-resistance to recorded data, is experiencing exponential growth in industry and research. In this paper, we propose the MIStore, a blockchain-based medical insurance storage system. Due to blockchain's the property of tamper-resistance, MIStore may provide a high-credibility to users. In a basic instance of the system, there are a hospital, patient, insurance company and n servers. Specifically, the hospital performs a (t, n)-threshold MIStore protocol among the n servers. For the protocol, any node of the blockchain may join the protocol to be a server if the node and the hospital wish. Patient's spending data is stored by the hospital in the blockchain and is protected by the n servers. Any t servers may help the insurance company to obtain a sum of a part of the patient's spending data, which servers can perform homomorphic computations on. However, the n servers cannot learn anything from the patient's spending data, which recorded in the blockchain, forever as long as more than n - t servers are honest. Besides, because most of verifications are performed by record-nodes and all related data is stored at the blockchain, thus the insurance company, servers and the hospital only need small memory and CPU. Finally, we deploy the MIStore on the Ethererum blockchain and give the corresponding performance evaluation.

Impact of COVID-19 pandemic on a world-wide private ophthalmic practice.

Purpose: To assess the impact of the COVID-19 pandemic on a worldwide private ophthalmic practice. Design: In this retrospective study, we reviewed the 2020 monthly outpatient and surgical volume of refractive, cataract, and retinal disease in Aier Eye clinics/hospitals of different regions, including the United States, Germany, Spain, Italy, and six major cities in China (Wuhan, Beijing, Shanghai, Shenyang, Urumqi and Yili). All of these data were compared to those of the same period of 2019. Results: Overall, during the early stage (2020 January to 2020 April) of COVID-19 outbreak, the outpatient and surgical volume of three main type ocular diseases (refractive, cataract and retinal surgery) showed an obvious reduction and reached the bottom in February in China. The data from the United States, Germany, Spain and Italy revealed the same trend, but the visit count nadir occurred until April, which is consistent with the spread trend of COVID-19 disease around the world. The average change rates of surgery volume (refractive, cataract and retinal surgery) in Chinese centers are 5.59%, -26.38%, 11.76%. The change rates of refractive (REF) and cataract volumes (CAT) in the United States are -8.62% and -10.58%, in Germany are -13.71% and -20.49%, in Spain are 15.35% and 27.97%, in Italy are 30.43% and -22.64%. In addition, the optometry outpatient volumes keep going up since May, with an average increasing rate of 21.18%, ranging from 7.43% to 49.51%. Conclusion: In conclusion, in this global chain of eye care units, the visit volumes of cataract, retinal and refractive changed significantly with the spread of COVID-19 pandemic. Among them, cataract surgery was the most affected sub-specialty, and refractive surgery and optometry volumes showed a potential growth in the near future. Therefore, medical institutions should make corresponding adjustments to the disease diagnosis and treatment strategies.

The thalamic covariance network is associated with cognitive deficits in patients with cerebral small vascular disease.

OBJECTIVE: Abnormalities in the gray matter structure of cerebral small vessel disease (CSVD) have been observed throughout the brain. However, whether cortico-cortical connections exist between regions of gray matter atrophy in patients with CSVD has not been fully elucidated. This question was tested by comparing the gray matter covariance networks in CSVD patients with and without cognitive impairment (CI). METHODS: We performed multivariate modeling of the gray matter volume measurements of 61 patients with CI (CSVD-CI), 85 patients without CI (CSVD-NC), and 108 healthy controls using source-based morphological analysis (SBM) to obtain gray matter structural covariance networks at the population level. Then, correlations between structural covariance networks and cognitive functions were analyzed in CSVD patients. Finally, a support vector machine (SVM) classifier was used with the gray matter covariance network as a classification feature to identify CI among the CSVD population. RESULTS: The results of the analysis of all the subjects showed that compared with healthy controls, the expression of the thalamic covariance network, cerebellum covariance network, and calcarine cortex covariance network was reduced in patients with CSVD. Moreover, CSVD-CI patients showed a significant reduction in the expression of the thalamic covariance network, encompassing the thalamus and the parahippocampal gyrus, relative to CSVD-NC patients, which persisted after excluding CSVD patients with thalamic lacunes. In patients with CSVD, cognitive functions were positively correlated with measures of the thalamic covariance network. More than 80% of CSVD patients with CI were correctly identified by the SVM classifier. INTERPRETATION: Our findings provide new evidence to explain the distribution state of gray matter reduction in CSVD patients, and the thalamic covariance network is the core region for early gray matter reduction during the development of CSVD disease, which is related to cognitive deficits. Reduced expression of thalamic covariance networks may provide a neuroimaging biomarker for the early identification of cognitive impairment in CSVD patients.

Intraocular lens constant optimization in toric intraocular lens calculation using keratometry and total corneal power.

PURPOSE: To evaluate intraocular lens (IOL) constant optimization in toric IOL calculation with keratometry (K) and total corneal refractive power (TCRP). METHODS: Predicted spherical equivalent (SE) and residual astigmatism (RA) with K and TCRP were retrospectively calculated using the Haigis, Holladay 1, and SRK/T formulae and optimized IOL constants. The results of the Barrett calculator and the Abulafia-Koch formula with K were also calculated. The median absolute error in SE (MedAE-SE), mean absolute error in RA (MAE-RA), and centroid error (CE) were analyzed. RESULTS: Seventy-nine eyes of 71 patients implanted with toric IOLs were included. With K, there were no significant differences between the results before and after constant optimization using all the formulae. With TCRP, constant optimization significantly reduced MedAE-SE; however, significantly increased MAE-RA and CE using the Holladay 1 and SRK/T formulae. MedAE-SE, MAE-RA, and CE using the Haigis formula did not show significant differences. The difference in the predicted RA before and after constant optimization increased with IOL toricity. The MedAE-SE predicted by TCRP was significantly higher than that predicted by K despite constant optimization. The MAE-RA and CE predicted by TCRP were significantly lower than those predicted by K without posterior corneal astigmatism optimization; however, were not significantly different from those predicted by the Barrett and Abulafia-Koch formulae. CONCLUSIONS: Constant optimization is recommended when using the TCRP in toric IOL calculations, particularly for patients with large astigmatism. However, TCRP did not yield more accurate results than optimized K in toric IOL calculations despite constant optimization.

Dynamic alterations in the amplitude of low-frequency fluctuation in patients with cerebral small vessel disease.

Background and purpose: Previous studies have focused on the changes of dynamic and static functional connections in cerebral small vessel disease (CSVD). However, the dynamic characteristics of local brain activity are poorly understood. The purpose of this study was to investigate the dynamic cerebral activity changes in patients with CSVD using the dynamic amplitude of low-frequency fluctuation (d-ALFF). Methods: A total of 104 CSVD patients with cognitive impairment (CSVD-CI, n = 52) or normal cognition (CSVD-NC, n = 52) and 63 matched healthy controls (HCs) were included in this study. Every participant underwent magnetic resonance imaging scans and a battery of neuropsychological examinations. The dynamics of spontaneous brain activity were assessed using dynamic changes in the amplitude of low-frequency fluctuation (ALFF) with the sliding-window method. We used voxel-wise one-way analysis of variance (ANOVA) to compare dynamic ALFF variability among the three groups. Post-hoc t-tests were used to evaluate differences between each group pair. Finally, the brain regions with d-ALFF values with differences between CSVD subgroups were taken as regions of interest (ROI), and the d-ALFF values corresponding to the ROI were extracted for partial correlation analysis with memory. Results: (1) There was no significant difference in age (p = 0.120), sex (p = 0.673) and education (p = 0.067) among CSVD-CI, CSVD-NC and HC groups, but there were significant differences Prevalence of hypertension and diabetes mellitus among the three groups (p < 10-3). There were significant differences in scores of several neuropsychological scales among the three groups (p < 10-3). (2) ANOVA and post-hoc t-test showed that there were dynamic abnormalities of spontaneous activity in several brain regions in three groups, mainly located in bilateral parahippocampal gyrus and bilateral hippocampus, bilateral insular and frontal lobes, and the static activity abnormalities in bilateral parahippocampal gyrus and bilateral hippocampal regions were observed at the same time, suggesting that bilateral parahippocampal gyrus and bilateral hippocampus may be the key brain regions for cognitive impairment caused by CSVD. (3) The correlation showed that d-ALFF in the bilateral insular was slightly correlated with the Mini-Mental State Examination (MMSE) score and disease progression rate. The d-ALFF value of the left postcentral gyrus was negatively correlated with the Clock Drawing Test (CDT) score (r = -0.416, p = 0.004), and the d-ALFF value of the right postcentral gyrus was negatively correlated with the Rey's Auditory Verbal Learning Test (RAVLT) word recognition (r = -0.320, p = 0.028). Conclusion: There is a wide range of dynamic abnormalities of spontaneous brain activity in patients with CSVD, in which the abnormalities of this activity in specific brain regions are related to memory and execution or emotion.

Vestibular Autorotation Test: The Differences in Peripheral and Central Acute Vestibular Syndrome.

Objective: To evaluate the difference between the vestibular autorotation test (VAT) in the peripheral and central acute vestibular syndrome (AVS). Methodology. Patients with AVS diagnosed by clinical manifestation admitted to the third affiliated hospital of Qiqihar Medical College from January 2019 to January 2021 were enrolled and divided into peripheral AVS (peripheral group) and central AVS (central group) according to the results of the MRI examination. Results: A total of 332 patients with AVS were recruited, including 282 patients in the peripheral group and 50 patients in the central group. The horizontal gain of both groups showed a downward trend at 2-6 Hz. There was no significant change in the horizontal phase between the two groups at 2-6 Hz. The horizontal gain of the two groups was stable at 2-6 Hz with no significant changes in the horizontal phase between 2-6 Hz in both groups. The central group showed a significantly lower proportion of gain increase coupled with loss and a strikingly higher proportion of gain increase without a loss than in the peripheral group (all P < 0.001). Conclusion: The increased horizontal and vertical gain of VAT in patients with AVS is of high value in the diagnosis of ACS. Significant differences in the results of VAT in patients with central and peripheral AVS could provide a reference for diagnosis.

Relationships Between Memory Impairments and Hippocampal Structure in Patients With Subcortical Ischemic Vascular Disease.

Background and Purpose: Patients with subcortical ischemic vascular disease (SIVD) suffer from memory disorders that are thought to be associated with the hippocampus. We aimed to explore changes in hippocampal subfields and the relationship between different hippocampal subfield volumes and different types of memory dysfunction in SIVD patients. Methods: A total of 77 SIVD patients with cognitive impairment (SIVD-CI, n = 39) or normal cognition (HC-SIVD, n = 38) and 41 matched healthy controls (HCs) were included in this study. Memory function was measured in all subjects, and structural magnetic resonance imaging (MRI) was performed. Then, the hippocampus was segmented and measured by FreeSurfer 6.0 software. One-way ANOVA was used to compare the volume of hippocampal subfields among the three groups while controlling for age, sex, education and intracranial volume (ICV). Then, post hoc tests were used to evaluate differences between each pair of groups. Finally, correlations between significantly different hippocampal subfield volumes and memory scores were tested in SIVD patients. Results: Almost all hippocampal subfields were significantly different among the three groups except for the bilateral hippocampal fissure (p = 0.366, p = 0.086, respectively.) and left parasubiculum (p = 0.166). Furthermore, the SIVD-CI patients showed smaller volumes in the right subiculum (p < 0.001), CA1 (p = 0.002), presubiculum (p = 0.002) and molecular layer of the hippocampus (p = 0.017) than the HC-SIVD patients. In addition, right subiculum volumes were positively related to Rey's Auditory Verbal Learning Test (RAVLT) word recognition (r = 0.230, p = 0.050), reverse digit span test (R-DST) (r = 0.326, p = 0.005) and Rey-Osterrieth Complex Figure Test (ROCF) immediate recall (r = 0.247, p = 0.035) scores, right CA1 volumes were positively correlated with RAVLT word recognition (r = 0.261, p = 0.026), and right presubiculum volumes showed positive relationships with R-DST (r = 0.254, p = 0.030) and ROCF immediate recall (r = 0.242, p = 0.039) scores. Conclusion: SIVD might lead to general reductions in volume in multiple hippocampal subfields. However, SIVD-CI patients showed atrophy in specific subfields, which might be associated with memory deficits.

Light-driven transformation of ZnS-cyclohexylamine nanocomposite into zinc hydroxysulfate: a photochemical route to inorganic nanosheets.

ZnS-CHA (CHA = cyclohexylamine) nanocomposite, a unique inorganic-organic hybrid semiconductor, has been prepared from a mild solvothermal reaction system. This material contains 2 nm-sized ZnS nanoparticles, and is photoactive toward UV light (≤300 nm). Under UV-irradiation, the ZnS-CHA nanocomposite is transformed to crystalline zinc hydroxysulfate nanosheets. The driving force of the light-driven transformation reaction is the photogeneration of charges (electrons and holes) in the ZnS nanoparticles, and these photogenerated charges interact with the CHA molecules and the inorganic S(2-) species, leading to decomposition of the organic component and self-oxidation of the inorganic ZnS nanoparticles to form zinc hydroxysulfate. Through simple thermal treatment, the as-formed zinc hydroxysulfate nanosheets are converted to ultrathin ZnO nanosheets with a porous feature, which exhibit high sensitivity and fast response and recovery for ethanol detection when used as an electrical sensing material.

Combining Calcitonin and Procalcitonin and Rheumatoid Arthritis-Related Biomarkers Improve Diagnostic Outcomes in Early Rheumatoid Arthritis.

OBJECTIVE: To demonstrate whether procalcitonin (PCT) combined with calcitonin (CT) could provide additional diagnostic value to other clinically available rheumatoid arthritis- (RA-) related biomarkers in the early diagnosis of RA. METHOD: The blood samples aseptically collected by venipuncture were centrifuged within 1 hour and frozen at -80°C. PCT and CT levels were measured using electrochemiluminescence immunoassay (ECLIA) in 260 subjects (48 patients with early RA, 34 patients with established RA, 37 patients with systemic lupus erythematosus, 30 with osteoarthritis, 31 with gouty arthritis, and 80 healthy participants). Anti-cyclic citrullinated peptide (Anti-CCP) and anti-RA33 antibodies (Anti-RA33) were analyzed by ELISA. RF was detected by transmission immunoturbidimetry. Mann-Whitney U tests and Kruskal-Wallis tests compared differences among groups. Spearman's rank correlation analysis determined the relationship between biomarkers. Receiver-operator characteristic (ROC) curves were generated, and diagnostic performance was assessed by area under the curve (AUC), as well as specificity, sensitivity, likelihood ratios (LR). RESULTS: Median serum PCT concentrations were significantly higher (p < 0.0001) in patients with early RA (0.065 ng/ml) when compared with healthy controls (0.024 ng/ml), and patients with osteoarthritis (0.025 ng/ml). When compared with gouty arthritis (GA) controls (0.072 ng/ml) and systemic lupus erythematosus (SLE) controls (0.093 ng/ml), median serum PCT concentrations were not significant in patients with early RA (0.065 ng/ml). Median serum CT concentrations were significantly lower (p < 0.0001) in patients with early RA (0.880 pg/ml) compared with healthy controls (3.159 pg/ml), patients with SLE (2.480 pg/ml), and patients with GA (2.550 pg/ml). When compared with osteoarthritis controls (0.586 pg/ml), median serum CT concentrations were not significant in patients with early RA (0.880 pg/ml). ROC curve analysis comparing early RA with healthy controls demonstrated that the AUC of RF, anti-CCP, and anti-RA33 were 0.66, 0.73, and 0.64, respectively; the additions of PCT and CT further improved the diagnostic ability of early RA with the AUC of 0.97, 0.98, and 0.97, respectively (p < 0.01). The sensitivities of RF, anti-CCP, and anti-RA33 for early RA were 33.33%, 44.74%, and 58.33%, respectively, and the additions of PCT and CT showed very high sensitivities of 83.33%, 92.11%, and 87.50%. The high-value groups of PCT moderately correlated with the anti-RA33 levels (r = 0.417, p < 0.05). CT had no significant correlation with disease duration, radiographic progression, or clinical/serological variables, such as ESR levels, CRP levels, RF, anti-CCP, and anti-RA33 levels in early RA. CONCLUSIONS: Serum PCT and CT combined with clinically available RA-related biomarkers could further improve the diagnostic efficiency of early RA.

Pleural Fluid GSDMD Is a Novel Biomarker for the Early Differential Diagnosis of Pleural Effusion.

OBJECTIVE: Gasdermin D (GSDMD), controlling pyroptosis in cells, has multiple physiological functions. The diagnostic role of GSDMD in pleural effusion (PE) remains unknown. METHODS: Sandwich ELISA kits that we developed were applied to measure the level of GSDMD for 335 patients with a definite cause of PE, including transudative PE, tuberculous pleural effusion (TPE), parapneumonic pleural effusion (PPE), and malignant pleural effusion (MPE). The diagnostic accuracy of Light's criteria vs. the new marker GSDMD was performed. Clinical follow-up of 40 cases of PPE was conducted and divided into efficacy and non-efficacy groups according to the therapeutic outcome. Nucleated cells (NCs) in PE were isolated and further infected with bacteria to verify the cell source of GSDMD. RESULTS: The diagnostic accuracy of GSDMD for the diagnosis of PE were 96% (sensitivity) and 94% (specificity). The receiver operating characteristic (ROC) curve indicated that GSDMD can be an efficient biomarker for the differential diagnosis of transudative PE and other groups (all AUC > 0.973). Noteworthily, the highest AUC belonged to tuberculosis diagnosis of 0.990, and the cut-off value was 18.40 ng/mL. Moreover, the same cut-off value of PPE and MPE was 9.35 ng/mL. The combination of GSDMD, adenosine deaminase (ADA), and lactate dehydrogenase (LDH) will further improve the diagnostic efficiency especially between TPE and PPE (AUC = 0.968). The AUC of GSDMD change at day 4, which could predict the therapeutic effect at an early stage, was 0.945 (P < 0.0001). Interestingly, bacterial infection experiments further confirm that the pleural fluid GSDMD was expressed and secreted mainly by the NCs. CONCLUSION: GSDMD and its combination are candidates as a potentially novel biomarker not only to separate PEs early and effectively, but also monitor disease progression.

Regional scalable priorities for national biodiversity and carbon conservation planning in Asia.

To achieve the goals of the post-2020 global biodiversity framework, we must identify representative targets that effectively protect biodiversity and can be implemented at a national level. We developed a framework to identify synergies between biodiversity and carbon across the Asian region and proposed a stepwise approach based on scalable priorities at regional, biome, and national levels that can complement potential Convention on Biological Diversity targets of protecting 30% land in the post-2020 global biodiversity framework. Our targets show that 30% of Asian land could effectively protect over 70% of all assessed species relative to only 11% now (based on analysis of 8932 terrestrial vertebrates), in addition to 2.3 to 3.6 hundred billion metric tons of carbon. Funding mechanisms are needed to ensure such targets to support biodiversity-carbon mutually beneficial solutions at the national level while reflecting broader priorities, especially in hyperdiverse countries where priorities exceed 30% of land.