Algorithm of automatic identification of diabetic retinopathy foci based on ultra-widefield scanning laser ophthalmoscopy.

AIM: To propose an algorithm for automatic detection of diabetic retinopathy (DR) lesions based on ultra-widefield scanning laser ophthalmoscopy (SLO). METHODS: The algorithm utilized the FasterRCNN (Faster Regions with CNN features)+ResNet50 (Residua Network 50)+FPN (Feature Pyramid Networks) method for detecting hemorrhagic spots, cotton wool spots, exudates, and microaneurysms in DR ultra-widefield SLO. Subimage segmentation combined with a deeper residual network FasterRCNN+ResNet50 was employed for feature extraction to enhance intelligent learning rate. Feature fusion was carried out by the feature pyramid network FPN, which significantly improved lesion detection rates in SLO fundus images. RESULTS: By analyzing 1076 ultra-widefield SLO images provided by our hospital, with a resolution of 2600×2048 dpi, the accuracy rates for hemorrhagic spots, cotton wool spots, exudates, and microaneurysms were found to be 87.23%, 83.57%, 86.75%, and 54.94%, respectively. CONCLUSION: The proposed algorithm demonstrates intelligent detection of DR lesions in ultra-widefield SLO, providing significant advantages over traditional fundus color imaging intelligent diagnosis algorithms.

Bezafibrate alleviates diabetes-induced spermatogenesis dysfunction by inhibiting inflammation and oxidative stress.

The metabolic disorders caused by diabetes can lead to various complications, including male spermatogenesis dysfunction. Exploring effective therapeutics that attenuate diabetes mellitus (DM)-induced male subfertility is of great importance. Pharmaceuticals targeting PPARα activation such as bezafibrate have been regarded as an important strategy for patients with diabetes. In this study, we use streptozocin (STZ) injection to establish a type 1 DM mice model and use bezafibrate to treat DM mice and evaluate the effects of bezafibrate on the spermatogenic function of the DM male mice. Bezafibrate treatment exhibited protective effects on DM-induced spermatogenesis deficiency, as reflected by increased testis weight, improved histological morphology of testis, elevated sperm parameters, increased serum testosterone concentration as well as increased mRNA levels of steroidogenesis enzymes. Meanwhile, testicular cell apoptosis, inflammation accumulation and oxidative stress status were also shown to be alleviated by bezafibrate compared with the DM group. In vivo and in vitro studies, PPARα specific inhibitor and PPARα knockout mice were further used to investigate the role of PPARα in the protective effects of bezafibrate on DM-induced spermatogenesis dysfunction. Our results indicated that the protection of bezafibrate on DM-induced spermatogenesis deficiency was abrogated by PPARα inhibition or deletion. Our study suggested that bezafibrate administration could ameliorate DM-induced spermatogenesis dysfunction and may represent a novel practical strategy for male infertility.

Learning to predict in-hospital mortality risk in the intensive care unit with attention-based temporal convolution network.

BACKGROUND: Dynamic prediction of patient mortality risk in the ICU with time series data is limited due to high dimensionality, uncertainty in sampling intervals, and other issues. A new deep learning method, temporal convolution network (TCN), makes it possible to deal with complex clinical time series data in ICU. We aimed to develop and validate it to predict mortality risk using time series data from MIMIC III dataset. METHODS: A total of 21,139 records of ICU stays were analysed and 17 physiological variables from the MIMIC III dataset were used to predict mortality risk. Then we compared the model performance of the attention-based TCN with that of traditional artificial intelligence (AI) methods. RESULTS: The area under receiver operating characteristic (AUCROC) and area under precision-recall curve (AUC-PR) of attention-based TCN for predicting the mortality risk 48 h after ICU admission were 0.837 (0.824 -0.850) and 0.454, respectively. The sensitivity and specificity of attention-based TCN were 67.1% and 82.6%, respectively, compared to the traditional AI method, which had a low sensitivity (< 50%). CONCLUSIONS: The attention-based TCN model achieved better performance in the prediction of mortality risk with time series data than traditional AI methods and conventional score-based models. The attention-based TCN mortality risk model has the potential for helping decision-making for critical patients. TRIAL REGISTRATION: Data used for the prediction of mortality risk were extracted from the freely accessible MIMIC III dataset. The project was approved by the Institutional Review Boards of Beth Israel Deaconess Medical Center (Boston, MA) and the Massachusetts Institute of Technology (Cambridge, MA). Requirement for individual patient consent was waived because the project did not impact clinical care and all protected health information was deidentified. The data were accessed via a data use agreement between PhysioNet, a National Institutes of Health-supported data repository (https://www.physionet.org/), and one of us (Yu-wen Chen, Certification Number: 28341490). All methods were carried out in accordance with the institutional guidelines and regulations.

A Simple and Quick Screening Method for Intrapulmonary Vascular Dilation in Cirrhotic Patients Based on Machine Learning.

BACKGROUND AND AIMS: Screening for hepatopulmonary syndrome in cirrhotic patients is limited due to the need to perform contrast enhanced echocardiography (CEE) and arterial blood gas (ABG) analysis. We aimed to develop a simple and quick method to screen for the presence of intrapulmonary vascular dilation (IPVD) using noninvasive and easily available variables with machine learning (ML) algorithms. METHODS: Cirrhotic patients were enrolled from our hospital. All eligible patients underwent CEE, ABG analysis and physical examination. We developed a two-step model based on three ML algorithms, namely, adaptive boosting (termed AdaBoost), gradient boosting decision tree (termed GBDT) and eXtreme gradient boosting (termed Xgboost). Noninvasive variables were input in the first step (the NI model), and for the second step (the NIBG model), a combination of noninvasive variables and ABG results were used. Model performance was determined by the area under the curve of receiver operating characteristics (AUCROCs), precision, recall, F1-score and accuracy. RESULTS: A total of 193 cirrhotic patients were ultimately analyzed. The AUCROCs of the NI and NIBG models were 0.850 (0.738-0.962) and 0.867 (0.760-0.973), respectively, and both had an accuracy of 87.2%. For both negative and positive cases, the recall values of the NI and NIBG models were both 0.867 (0.760-0.973) and 0.875 (0.771-0.979), respectively, and the precisions were 0.813 (0.690-0.935) and 0.913 (0.825-1.000), respectively. CONCLUSIONS: We developed a two-step model based on ML using noninvasive variables and ABG results to screen for the presence of IPVD in cirrhotic patients. This model may partly solve the problem of limited access to CEE and ABG by a large numbers of cirrhotic patients.

Clinical Performance of the Xpert® CT/NG Test for Detection of Chlamydia trachomatis and Neisseria gonorrhoeae: A Multicenter Evaluation in Chinese Urban Hospitals.

Background: We aimed to evaluate the clinical performance of the GeneXpert® (Xpert) CT/NG assay for the detection of Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) using urine and cervical swabs collected from patients in China. Methods: This study was conducted from September 2016 to September 2018 in three Chinese urban hospitals. The results from the Xpert CT/NG test were compared to those from the Roche cobas® 4800 CT/NG test. Discordant results were confirmed by DNA sequence analysis. Results: In this study, 619 first void urine (FVU) specimens and 1,042 cervical swab specimens were included in the final dataset. There were no statistical differences between the results of the two tests for the detection of CT/NG in urine samples (p > 0.05), while a statistical difference was found in cervical swabs (p < 0.05). For CT detection, the sensitivity and specificity of the Xpert test were 100.0% (95%CI = 96.8-99.9) and 98.3% (95%CI = 96.6-99.2) for urine samples and 99.4% (95%CI = 96.5-100.0) and 98.6% (95%CI 97.5-99.2) for cervical swabs, respectively. For NG detection, the sensitivity and specificity of the Xpert test were 99.2% (95%CI = 94.9-100.0) and 100.0% (95%CI = 99.0-100.0) for urine and 100% (95%CI = 92.8-100.0) and 99.7% (95%CI = 99.0-99.9) for cervical swabs, respectively. Conclusion: The Xpert CT/NG test exhibited high sensitivity and specificity in the detection of CT and NG in both urine and cervical samples when compared to the reference results. The 90-min turnaround time for CT and NG detection at the point of care using Xpert may enable patients to receive treatment promptly.

A better method for the dynamic, precise estimating of blood/haemoglobin loss based on deep learning of artificial intelligence.

BACKGROUND: Dynamic and precise estimation of blood loss (EBL) is quite important for perioperative management. To date, the Triton System, based on feature extraction technology (FET), has been applied to estimate intra-operative haemoglobin (Hb) loss but is unable to directly assess the amount of blood loss. We aimed to develop a method for the dynamic and precise EBL and estimate Hb loss (EHL) based on artificial intelligence (AI). METHODS: We collected surgical patients' non-recycled blood to generate blood-soaked sponges at a set gradient of volume. After image acquisition and preprocessing, FET and densely connected convolutional networks (DenseNet) were applied for EBL and EHL. The accuracy was evaluated using R2, the mean absolute error (MAE), the mean square error (MSE), and the Bland-Altman analysis. RESULTS: For EBL, the R2, MAE and MSE for the method based on DenseNet were 0.966 (95% CI: 0.962-0.971), 0.186 (95% CI: 0.167-0.207) and 0.096 (95% CI: 0.084-0.109), respectively. For EHL, the R2, MAE and MSE for the method based on DenseNet were 0.941 (95% CI: 0.934-0.948), 0.325 (95% CI: 0.293-0.355) and 0.284 (95% CI: 0.251-0.317), respectively. The accuracies of EBL and EHL based on DenseNet were more satisfactory than that of FET. Bland-Altman analysis revealed a bias of 0.02 ml with narrow limits of agreement (LOA) (-0.47 to 0.52 mL) and of 0.05 g with narrow LOA (-0.87 to 0.97 g) between the methods based on DenseNet and actual blood loss and Hb loss. CONCLUSIONS: We developed a simpler and more accurate AI-based method for EBL and EHL, which may be more fit for surgeries primarily using sponges and with a small to medium amount of blood loss.

Comparison of Microdilution Method with Agar Dilution Method for Antibiotic Susceptibility Test of Neisseria gonorrhoeae.

INTRODUCTION: Antimicrobial resistance (AMR) of Neisseria gonorrhoeae (N. gonorrhoeae) becomes a grave public health problem in the world. A strengthened Antimicrobial Resistance Surveillance Program is needed to track the trend of AMR development. However, the lack of a proper antimicrobial susceptibility test (AST) method is a barrier to expand the AMR surveillance in China. Traditional agar dilution (AD) method is laborious and E-test strips have no approval license for clinical use. Herein, a Chinese group modified the microdilution (MD) method for clinical ASTs. The objective of this study is to compare the MD method with the AD method for N. gonorrhoeae AST. MATERIALS AND METHODS: A total of 166 clinical isolates were tested for antimicrobial susceptibility of ceftriaxone, spectinomycin, azithromycin, ciprofloxacin, tetracycline, and penicillin using MD and AD method simultaneously. Results of MD method were read manually or automatically. Rates of essential agreement (EA), category agreement (CA), minor error, and very major error were compared. RESULTS: The total EAs (compared with results read manually) of penicillin, tetracycline, ciprofloxacin, spectinomycin, ceftriaxone, and azithromycin were 90.4%, 97.0%, 85.5%, 100.0%, 94%, and 72.3%; and CAs were 82.5%, 94.0%, 100%, 100%, 95.2%, and 94%, respectively. CONCLUSION: We conclude that the MD method might be an alternative for clinical AST of N. gonorrhoeae in China. In particular, MD method has the potency of accurate differentiation of isolates resistant to ceftriaxone or azithromycin, which were empirically recommended for gonococcal treatment, but its quality remained suboptimal, and further improvement is needed for clinical use.

Identification and expression analysis of ceftriaxone resistance-related genes in Neisseria gonorrhoeae integrating RNA-Seq data and qRT-PCR validation.

OBJECTIVES: The aim of the study is to identify ceftriaxone resistance-related genes in Neisseria gonorrhoeae. METHODS: Differences in gene expression were compared between ceftriaxone-susceptible N. gonorrhoeae isolates [minimum inhibitory concentration (MIC)=0.002-0.004mg/L] and isolates with decreased ceftriaxone susceptibility (MIC=0.125-0.5mg/L) using RNA-Seq (RNA sequencing). RESULTS: Total RNA of 10 clinical isolates was used to make libraries and generated an average of 24.07Mb reads per sample; these were assembled into 1871 mRNA genes. Moreover, 21 differentially expressed genes (DEGs) were found between the N. gonorrhoeae isolates with susceptibility and decreased susceptibility to ceftriaxone with a fold change of ≥2 (P<0.05), among which 11 were upregulated and 10 were downregulated. Furthermore, all DEGs were verified by quantitative reverse transcription PCR (qRT-PCR), which detected 25 clinical isolates with decreased ceftriaxone susceptibility and 21 ceftriaxone-susceptible isolates. In addition, seven DEGs revealed relative expression levels by 2-ΔΔCt and showed a statistical significance (P≤0.05). Analysis of Gene Ontology (GO) terms and KEGG pathway for functional enrichment showed that six DEGs were related to the cellular component and one DEG was related to the biosynthesis of antibiotics, and these results might be related to ceftriaxone resistance. CONCLUSIONS: Examining ceftriaxone resistance-related genes in N. gonorrhoeae is necessary owing to the high morbidity and antimicrobial resistance of N. gonorrhoeae, especially its eventual resistance to third-generation extended-spectrum cephalosporins (cefixime and ceftriaxone). Moreover, this report provides a new direction for the study and control of ceftriaxone-resistant N. gonorrhoeae.

catena-Poly[[(1,3-dimethyl-imidazolin-2-one-κN)(1,10-phenanthroline-κ(2)N,N')copper(II)]-µ-furan-2,5-dicarboxyl-ato-κ(2)O(2):O(5)].

The polymeric title compound, [Cu(C(6)H(2)O(5))(C(12)H(8)N(2))(C(5)H(10)N(2)O)](n), is composed of an infinite chain formed along [100] by linking the (1,3-dimethyl-imidazolin-2-one)(1,10-phenanthroline)copper(II) units with two O atoms of two carboxyl-ate groups of the furan-2,5-dicarboxyl-ate ligand. The Cu(II) atom, which lies on a twofold rotation axis, displays a square-pyramidal coordination. The dihedral angles of the 1,10-phenanthroline ligand with respect to the furan rings of the carboxyl-ate anions that are connected to the metal atom are 62.18 (11) and 88.27 (12)°.

catena-Poly[[(1,10-phenanthroline-κ(2)N,N')zinc]-µ-furan-2,5-dicarboxyl-ato-κ(4)O(2),O(2'):O(5),O(5')].

In the title coordination polymer, [Zn(C(6)H(2)O(5))(C(12)H(8)N(2))](n), an infinite chain is formed along [010] by linking the chelated {Zn(phen)} entities (phen is 1,10-phenanthroline) with two carboxyl-ate groups of the furan-2,5-dicarboxyl-ate ligand. The Zn(II) atom shows trigonal-prismatic coordination.

catena-Poly[[triaquacopper(II)]-µ(2)-furan-2,5-dicarboxyl-ato-κ(4)O(1),O(2):O(2),O(2')].

In the title compound, [Cu(C(6)H(2)O(5))(H(2)O)(3)](n), an infinite chain is formed along [001] by linking of the Cu(OH(2))(3)O(4) cluster with one carboxyl-ate group of the furan-2,5-dicarboxyl-ate ligand. Adjacent chains are linked by O(water)-H⋯O hydrogen-bonding inter-actions. The Cu(OH(2))(3)O(4) cluster displays a penta-gonal bipyrimadal geometry with two weak coordinations [Cu-O(furan) = 2.790 (2) Å) and Cu-O(carboxyl-ate) = 2.684 (2) Å] and two water mol-ecules located in axial positions.

catena-Poly[(triaqua-zinc)-µ-furan-2,5-dicarboxyl-ato-κ(3)O(2):O(2),O(2')].

In the crystal structure of the title compound, [Zn(C(6)H(2)O(5))(H(2)O)(3)](n), an infinite chain is formed along [001] by linking of the Zn(H(2)O)(3) entities with one carboxyl-ate group of the furan-2,5-dicarboxyl-ate ligand. Adjacent chains are linked by O(water)-H⋯O hydrogen-bonding inter-actions. The Zn(H(2)O)(3)O(3) polyhedron displays a distorted octa-hedral geometry with one weak Zn-O(carboxyl-ate) coordination [2.433 (8) A°] and two water mol-ecules located in axial positions. Except for one of the axial water molecules and two adjacent H atoms, the other atoms (including H atoms) possess site symmetry m.

catena-Poly[(aqua-dimethano-lzinc)-µ-furan-2,5-dicarboxyl-ato-κ(3)O(2):O(2),O(2')].

In the crystal structure of the title compound, [Zn(C(6)H(2)O(5))(CH(3)OH)(2)(H(2)O)](n), an infinite chain is formed along the b axis by linking of the Zn(OH(2))(CH(3)OH)(2) unit with one carboxyl-ate group of the furan-2,5-dicarboxyl-ate ligand. The Zn(II) ion is in a distorted octa-hedral environment with one weak coordination [Zn-O(carboxyl-ate) = 2.565 (3) Å] and two meth-anol mol-ecules located in axial positions. In the chain, O(water)-H⋯O hydrogen bonds are present, while adjacent chains are linked by O(methanol)-H⋯O hydrogen bonds into a layer parallel to (10-2).

catena-Poly[[[aqua-bis-(1H-imidazole-κN(3))copper(II)]-µ-furan-2,5-di-car-boxylato-κ(2)O(2):O(5)] trihydrate].

In the title cooridnation polymer, {[Cu(C(6)H(2)O(5))(C(3)H(4)N(2))(2)(H(2)O)]·3H(2)O}(n), an infinite chain is formed along [001] by linking of the Cu(C(3)N(2)H(4))(2)(H(2)O) entities with two bridging monodentate carboxyl-ate groups of two different furan-2,5-dicarboxyl-ate dianions. The geometry of the Cu(2+) ion is a square-based pyramid with the water atom in the apical position and the ligand O and N atoms in a trans orientation. The dihedral angle between the imidazole planes is 83.96 (14)°. O(w)-H⋯O and N(i)-H⋯O (w = water and i = imidazole) hydrogen bonds help to establish the packing.

[Expression, purification and activity identification of ZtaN-p23 fusion protein in Escherichia coli of Epstein-Barr virus].

AIM: To construct the prokaryotic expression plasmid pGEX-4T-1-BZLF1N-BLRF2, and express it in Escherichia coli. METHODS: The EB virus BZLF1N gene and BLRF2 gene were amplified by RT-PCR respectively. Then, the two genes were linked by splicing overlap extension PCR method and inserted into the vector pGEX-4T-1, and the recombinant plasmid pGEX-4T-1-BZLF1N-BLRF2 was transformed into E.coli BL21 (DE3) strain. The expression protein ZtaN-p23 was analysed by SDS-PAGE and immunoreactivity was proved by Western blotting. RESULTS: Restriction enzyme digestion and DNA sequencing showed recombinant plasmid constructed successfully. The expression product ZtaN-p23 with the molecular weight 46000 was located in the cytoplasm and insoluble. The ZtaN-p23 up to 95% purity was obtained after purified using affinity chromatography. Western blotting showed fusion protein possessed a well bioactivity and specificity. CONCLUSION: The fusion gene BZLF1N-BLRF2 is successfully constructed and effectively expressed in E.coli, which lay the foundation for further research on its biological properties and functions.

catena-Poly[[[aqua-(2,2'-bipyridine-κ(2) N,N')zinc]-µ-furan-2,5-dicarboxyl-ato-κ(2) O (2):O (5)] dihydrate].

In the title hydrated coordination polymer, {[Zn(C6H2O5)(C10H8N2)(H2O)]·2H2O} n , an infinite [1-10] chain is formed by the linking of [Zn(C10H8N2)(H2O)](2+) entities by bridging, monodentate furan-2,5-dicarboxyl-ate dianionic linkers. The Zn(2+) coordination geometry is a trigonal bipyramid, with one N atom (from 2,2'-bipyridine) and one O atom (from the bridging dianion) in the axial positions. For each Zn(II) atom, the dihedral angle between the furan ring of its coordinated bridging ligand and its coordinated bipyridine ring system is 87.19 (8)°. O-H⋯O hydrogen bonds involving both the coordinated and uncoordinated water mol-ecules generate a layer motif parallel to (001).

Poly[[µ-aqua-diaqua-bis-(µ-furan-2,5-dicarboxyl-ato-κ(2) O (2):O (5))bis-(1,10-phenanthroline-κ(2) N,N')dicopper(II)] N,N-dimethyl-formamide monosolvate].

The asymmetric unit of the title compound, {[Cu2(C6H2O5)2(C12H8N2)2(H2O)3]·C3H7NO} n , contains two Cu(II) atoms, two furan-2,5-dicarboxyl-ate (L) ligands, two 1,10-phenanthroline (phen) ligands, three coordinating water mol-ecules and one N,N-dimethyl-formamide solvent mol-ecule. Each Cu(II) atom is coordinated by two N atoms from one phen ligand, two O atoms from two L ligands and two water mol-ecules in a distorted octa-hedral geometry. The main difference between the environments of the two independent Cu atoms is in the Cu-Owater distances, which are 2.415 (2) and 2.639 (2) Šfor one Cu(II) atom and 2.3560 (19) and 2.911 (4) Šfor the other. Ligands L and one independent water mol-ecule bridge the Cu(II) atoms, forming corrugated polymeric layers parallel to the ab plane. Inter-molecular O-H⋯O and C-H⋯O hydrogen bonds consolidate the crystal packing.

catena-Poly[[[aqua-(2,2'-bipyridine-κ(2)N,N')copper(II)]-µ-furan-2,5-di-carboxyl-ato-κ(2)O(2):O(5)] dihydrate].

In the crystal structure of the title compound, {[Cu(C(6)H(2)O(5))(C(10)H(8)N(2))(H(2)O)]·2H(2)O}(n), an infinite chain parallel to [110] is formed by the linking of Cu(H(2)O)(2,2'-bipyridine) units through a furan-2,5-dicarboxyl-ate bridge. The Cu(II) atom shows a square-pyramidal geometry, with one furan-2,5-dicarboxyl-ate O atom in the apical position. The dihedral angle between the planes of the furan ring and the bipyridine mol-ecule is 83.88 (7)°. O(water)-H⋯O hydrogen bonds connect adjacent chains, generating a layer motif parallel to (001).

[Cloning and expression of human sPD-1-Fc in Cos-7 cells].

AIM: To investigate human soluble PD-1 (sPD-1) biological activity sPD-1 gene be cloned and expressed in eucaryote cells. METHODS: sPD-1 gene was amplified from human PBMC mRNA by RT-PCR and cloned into eucaryotic expression vector pSG5-Fc. And the positive recombinant plasmid pSG5-Fc -sPD-1 was screened by enzyme digestion and sequencing.The correct sequencing of the recombinant plasmid pSG5-Fc-sPD-1 was transfected into COS-7 cells. The expressed recombinant protein in the supernatant was concentrated with protein A-agarose and analyzed by Western blot. The binding activity to PD-L1 which was expressed with prokaryotic cells was detected with Co-IP. RESULTS: The human sPD-1 fragment was obtained through RT-PCR. The plasmid pSG5-Fc-sPD-1 was constructed by double enzyme digestion and ligation of vector pSG5-Fc and fragment sPD-1. Sequenced sPD-1 gene was coincident with the theoretical sequence. sPD-1-Fc fusion protein in the supernatant was expressed in COS-7 cells and identified by Western blot. The activity of recombinant fusion protein sPD-1-Fc bound to PD-L1 had been detected with Co-IP. CONCLUSION: The human sPD-1 has been cloned and expressed in eucaryote cells successfully. The sPD-1-Fc fusion protein can be effective in binding to PD-L1, which can be used for further research in the function and clinical application of sPD-1.

[Effects of grain size change on landscape pattern indices of Hefei City].

Based on the land use map of Hefei in 2002, and by using ArcView software and Fragstats 3.3 spatial analysis program, this paper studied the change characteristics of selected 22 landscape pattern indices of Hefei under effects of different grain size, and analyzed the fitting functions and variation coefficients of partial landscape indices. The results showed that with the grain size increased from 10 to 100 m, the selected indices displayed different change trend, illustrating that the change of grain size had obvious effects on landscape indices. The effects of the grain size could be classified as 4 kinds of situation, i.e., overall rise, overall drop, less change, and no regulation change. The response curves of landscape pattern indices to grain size were fitted by quadratic function, cubic function, power function, and linear function, respectively. For the 1:50000 land use map of Hefei, the suitable range of grain size for computing and analyzing landscape indices was 15-35 m.