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Abstract Background Global myopia prevalence poses a substantial public health burden with vision-threatening complications, necessitating effective prevention and control strategies. Precise prediction of spherical equivalent (SE), myopia, and high myopia onset is vital for proactive clinical interventions. Methods We reviewed electronic medical records of pediatric and adolescent patients who underwent cycloplegic refraction measurements at the Eye & Ear, Nose, and Throat Hospital of Fudan University between January 2005 and December 2019. Patients aged 3–18 years who met the inclusion criteria were enrolled in this study. To predict the SE and onset of myopia and high myopia in a specific year, two distinct models, random forest (RF) and the gradient boosted tree algorithm (XGBoost), were trained and validated based on variables such as age at baseline, and SE at various intervals. Outputs included SE, the onset of myopia, and high myopia up to 15 years post-initial examination. Age-stratified analyses and feature importance assessments were conducted to augment the clinical significance of the models. Results The study enrolled 88,250 individuals with 408,255 refraction records. The XGBoost-based SE prediction model consistently demonstrated robust and better performance than RF over 15 years, maintaining an R 2 exceeding 0.729, and a Mean Absolute Error ranging from 0.078 to 1.802 in the test set. Myopia onset prediction exhibited strong area under the curve (AUC) values between 0.845 and 0.953 over 15 years, and high myopia onset prediction showed robust AUC values (0.807–0.997 over 13 years, with the 14th year at 0.765), emphasizing the models' effectiveness across age groups and temporal dimensions on the test set. Additionally, our classification models exhibited excellent calibration, as evidenced by consistently low brier score values, all falling below 0.25. Moreover, our findings underscore the importance of commencing regular examinations at an early age to predict high myopia. Conclusions The XGBoost predictive models exhibited high accuracy in predicting SE, onset of myopia, and high myopia among children and adolescents aged 3–18 years. Our findings emphasize the importance of early and regular examinations at a young age for predicting high myopia, thereby providing valuable insights for clinical practice.

Abstract Background This study aimed to evaluate the clinical outcomes and assess preoperative characteristics that may predict outcomes in keratoconus 1 year after accelerated transepithelial corneal cross-linking (ATE-CXL). Methods This prospective study included 93 eyes of 84 consecutive keratoconus patients with 1-year follow-up after ATE-CXL. Preoperative characteristics included corneal astigmatism, anterior chamber depth, anterior chamber volume, radius of curvature, posterior elevation, central corneal thickness (CCT), thinnest corneal thickness, steepest meridian keratometry, flattest meridian keratometry, and the maximum keratometry (Kmax). Data were obtained preoperatively and at 1, 3, 6, and 12 months postoperatively. The patient eyes were grouped into 3 subgroups according to CCT and Kmax values to observe the changes of keratoconus progression. Results All patients were successfully operated without complications at any follow-up time point. Mean changes of Kmax from baseline at 6 and 12 months were − 0.60 ± 2.21 D (P = 0.011) and − 0.36 ± 1.58 D (P = 0.030), respectively. Eyes with a thinner CCT and higher Kmax values exhibited a tendency for topographic flattening of ≥1.0 D (P = 0.003; P = 0.003). In the subgroup comparison, the Kmax values decreased significantly at 6 and 12 months after ATE-CXL in the group with CCT ≤ 450 μm (P = 0.018 and P = 0.045); the Kmax values of the group with Kmax > 65.0 D decreased significantly at 6 months postoperatively (P = 0.025). Conclusion ATE-CXL is a safe and effective treatment for keratoconus patients. Patients with thinner CCT and higher Kmax values are more likely to benefit from ATE-CXL.

African swine fever (ASF) is a highly fatal porcine disease caused by the African swine fever virus (ASFV), and resulting in huge economic losses across the globe. ASF has been raging in China for 3 years, and recently EP402R-deleted ASFV strains emerged, showing sub-acute or chronic symptoms in pigs and providing novel difficulties to monitor and control the disease as EP402R-deleted strains possess no hemadsorption (HAD) ability. In addition, the gene deletion virus with low viral load is prone to results retest or false negative due to the high cycle threshold (Ct) value under the current real-time polymerase chain reaction (PCR) detection method. Thus, a new method is needed to detect and distinguish wild strains and gene-deleted viruses. In this study, a duplex droplet digital polymerase chain reaction (ddPCR) assay based on the ASFV B646L and EP402R genes was established and showed good linearity (R2 > 0.99). The limit of detection for duplex ddPCR was 52 copies per reaction and 8.6 copies per reaction for B646L and EP402R, respectively. No cross-reaction with other porcine viruses [classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV), porcine parvovirus (PPV), Japanese encephalitis virus (JEV), and porcine circovirus type 2 (PCV2)] was identified by this assay. In addition, 44 ASFV-suspicious clinical samples as well as EP402R-deleted ASFV were tested in parallel by duplex real-time PCR and ddPCR, indicative of a higher sensitivity which belonged to the duplex ddPCR assay. In summary, this is the first time that duplex ddPCR assay has been successfully developed to provide an efficient method to detect and differentiate ASFV wild-type and gene-deleted strains.

Background: Keratectasia after corneal refractive surgery is a rare but serious postoperative complication, and reports on accelerated transepithelial corneal crosslinking (ATE-CXL)-based treatment of patients with post-laser-assisted in situ keratomileusis (LASIK) ectasia are limited. Therefore, this study evaluated the long-term efficacy and safety of ATE-CXL for progressive post-LASIK ectasia.Methods: This prospective observational study was conducted at the Eye and ENT Hospital, Fudan University, Shanghai, China, and 25 eyes from 25 patients with post-LASIK ectasia undergoing ATE-CXL were examined. Clinical examinations were conducted preoperatively and postoperatively to assess parameters such as manifest refraction, corrected distance visual acuity (CDVA), endothelial cell density; keratometry, corneal thickness, posterior elevation and topometric indices were measured using Pentacam; sectoral pachymetry and epithelial thickness were evaluated using optical coherence tomography. A paired t-test, Wilcoxon rank-sum test, Kruskal-Wallis test, and repeated measures analysis of variance were used for statistical analysis.Results: Participants were examined for an average of 46 months. No severe complications occurred during or after ATE-CXL. CDVA improved from 0.25 ± 0.31 preoperatively to 0.15 ± 0.17 postoperatively (p = 0.011). Maximum keratometry decreased from 55.20 ± 8.33 D to 54.40 ± 7.98 D, with no statistical significance (p = 0.074), and the central corneal thickness increased from 414.92 ± 40.96 μm to 420.28 ± 44.78 μm (p = 0.047) at the final follow-up. Posterior elevation, pachymetry, and epithelial thickness remained stable (p > 0.05) throughout the follow-up. No significant differences were noted in topometric indices, except the central keratoconus index, which decreased significantly (p < 0.001) at the final follow-up.Conclusion: Improvements in CDVA and stabilization in corneal keratometry and posterior elevation after ATE-CXL were noted at the 46-months follow-up, demonstrating that ATE-CXL is a safe and effective treatment for progressive post-LASIK ectasia.

African swine fever virus (ASFV) causes a viral disease in swine with a mortality rate of approximately 100%, threatening the global pig industry’s economic development. However, vaccines are not yet commercially available, and other antiviral therapeutics, such as antiviral drugs, are urgently needed. In this study, berbamine hydrochloride, a natural bis-benzylisoquinoline alkaloid isolated from the traditional Chinese herb Berberis amurensis, showed significant antiviral activity against ASFV. The 50% cytotoxic concentration (CC50) of berbamine hydrochloride in porcine alveolar macrophages (PAMs) was 27.89 μM. The antiviral activity assay demonstrated that berbamine hydrochloride inhibits ASFV in a dose-dependent manner. In addition, a 4.14 log TCID50 decrease in the viral titre resulting from non-cytotoxic berbamine hydrochloride was found. Moreover, the antiviral activity of berbamine hydrochloride was maintained for 48h and took effect at multiplicities of infection (MOI) of 0.01, 0.1, and 1. The time-of-addition analysis revealed an inhibitory effect throughout the entire virus life-cycle. A subsequent viral entry assay verified that berbamine hydrochloride blocks the early stage of ASFV infection. Moreover, similar anti-ASFV activity of berbamine hydrochloride was also found in PK-15 and 3D4/21 cells. In summary, these results indicate that berbamine hydrochloride is an effective anti-ASFV natural product and may be considered a novel antiviral drug.

Aims: This study aimed to investigate the corneal biomechanical changes and topographic outcomes of accelerated transepithelial corneal cross-linking (ATE-CXL) in pediatric progressive keratoconus.Methods: In this prospective longitudinal study, 31 eyes of 28 pediatric patients with keratoconus (21 boys and 7 girls; mean age, 14.35 ± 2.68 years) undergoing ATE-CXL (epithelium-on procedure with 45 mW/cm2 for 320 s) were included. Corvis ST was used to measure dynamic corneal response parameters at baseline and at 12 month after ATE-CXL. Corneal keratometry and corneal thickness were measured using Pentacam pre-operatively and 1, 6, and 12 month post-operatively.Results: No serious complications occurred during or after ATE-CXL. The maximum keratometry values were 60.10 ± 7.51 D pre-operatively and 61.42 ± 8.92, 61.17 ± 7.96, and 60.02 ± 7.58 D at 1, 6, and 12 month after ATE-CXL (P > 0.05), respectively. Corneal thickness remained stable during the 12-month follow-up (P > 0.05). At post-operative 12 month, first applanation time (P < 0.001), first applanation length (P = 0.004), second applanation velocity (P = 0.014), highest concavity time (P = 0.022), and radius of curvature at highest concavity (P = 0.031) increased significantly. The value of stiffness parameter at first applanation was significantly increased from 57.70 ± 27.57 pre-operatively to 63.36 ± 27.09 at 12 months after ATE-CXL (P = 0.018).Conclusions: ATE-CXL is safe and effective in stabilizing the progression of pediatric keratoconus. Changes in corneal biomechanical response consistent with stiffening following ATE-CXL were observed in pediatric patients with keratoconus.

Abstract Background Conventional corneal cross-linking is effective for retarding the progression of keratoconus. However the long-term efficacy and safety of accelerated (45 mW/cm2) transepithelial corneal cross-linking (ATE-CXL) on progressive keratoconus (KC) treatment is not fully understood. The purpose of this study is to evaluate the 2-year changes in corneal topographic parameters and densitometry values after ATE-CXL for KC. Methods Twenty-five progressive eyes of 25 KC patients (KC group) and 25 eyes of 25 myopes without KC (control group) were enrolled. Corneal topography and densitometry values were evaluated pre-operatively and at 6, 12 and 24 months post-operatively in the KC group. Results The mean values of flat keratometry (K1), steep keratometry (K2), mean keratometry (Km), corneal astigmatism (CA), maximum keratometry (Kmax), central corneal thickness (CCT), thinnest corneal thickness (TCT), anterior corneal elevation (ACE) and posterior corneal elevation (PCE) all remained unchanged over time (all P values > 0.05). The densitometry values of the anterior, central, posterior and total layers over the annular diameters 0 mm to 2 mm (Φ0-2 mm) and Φ2–6 mm all decreased significantly (all P values 0.05). Subgroups with Km ≥ 50.30D or ACE ≥35.3 μm progressed significantly when compared with those with Km

Purpose. To compare the corneal biomechanical properties between post-LASIK ectasia and primary keratoconus. Methods. A total of 42 eyes of 42 patients with matching age and central corneal thickness (CCT) were divided into two groups according to diagnosis of post-LASIK ectasia (PLE group; n = 21; age range: 22–47 years) and primary keratoconus (KC group; n = 21; age range: 21–47 years). The corneal biomechanical properties were assessed using Scheimpflug-based technology (Corvis ST; Oculus Optikgeräte, Wetzlar, Germany). The paired t-test and linear regression analysis were performed. Results. The PLE group had significantly higher mean stiffness parameter at the first applanation (SP-A1; 76.65 ± 21.66 vs 52.72 ± 13.65, p≤0.001) and mean stress-strain index (SSI) (SSI: 0.78 ± 0.16 versus 0.64 ± 0.12, p=0.001) than the KC group. SP-A1 was positively correlated with CCT in the PLE group (Pearson’s r = 0.816, p≤0.001), but not in the KC group (Pearson’s r = −0.014, p=0.952). No statistical correlation was observed between SSI and CCT in either group (Pearson’s r = 0.292, p=0.199, and Pearson’s r = 0.004, p=0.985, respectively). Conclusions. In our case series, KC manifested more severe than PLE in biomechanical properties. Since SSI measurements were independent of corneal thickness, it can be used for corneal biomechanical assessment.

Abstract Background Keratoconus typically presents in the teenage years and is more advanced in younger patients when compared with adults. In the present study, we aimed to assess the safety and efficacy of accelerated transepithelial corneal collagen cross-linking (ATE-CXL) in children with progressive keratoconus. Methods In this retrospective consecutive study, 18 eyes were enrolled from 17 pediatric patients (15 boys and 2 girls) with a mean age of 14.44 ± 1.98 years. Manifest refraction, best-corrected visual acuity (BCVA), steepest meridian keratometry (K1), flattest meridian keratometry (K2), maximum keratometry (Kmax), thinnest corneal thickness (TCT), posterior central elevation (PCE), and posterior mean elevation (PME) were measured before and after ATE-CXL. The patients were followed-up at 1, 6, and 12 months. Repeated measures analysis of variance was used for statistical analysis. P 0.05). Conclusions ATE-CXL is a safe and effective treatment in pediatric progressive keratoconus patients. The long-term effects need further observation. Trial registration Retrospectively registered. Registration number: ChiCTR-OIC-16008181. Registered 29 March 2016.

Abstract Background Adolescent KC is a special segment of the general KC population because an adolescents’s eyes are still susceptible to blurred vision and optical defocus during the sensitive period of ocular and visual development. In the present study, we evaluated the ocular dimensions of 53 KC adolescents. Methods One hundred and six KC eyes of 53 (42 boys and 11 girls) KC adolescents (age 15.5 ± 1.7 years, range 11 to 18) were involved in this retrospective study. The eye with more severe KC (Severe Group) of each patient was compared with their less affected eye (Mild Group). Optical axial length (OAL) was measured by optical coherence biometry (IOL-master). Central corneal thickness, anterior chamber depth (ACD), flat keratometry value, steep keratometry value, and maximum keratometry value were assessed with an anterior segment analyzer (Pentacam HR). Anterior segment length (ASL) was manually measured from the 25 scheimpflug images captured by the Pentacam HR with the mean value recorded. The posterior segment length (PSL) was calculated with the formula “PSL = OAL-ASL”. Results The mean ACD, OAL, ASL, and PSL values of the Severe Group were 3.51 ± 0.32 mm, 24.76 ± 1.24 mm, 4.01 ± 0.30 mm and 20.76 ± 1.15 mm.While those of the Mild Group were 3.36 ± 0.29 mm, 24.97 ± 1.40 mm, 3.94 ± 0.35 mm and 21.03 ± 1.31 mm. The Severe Group has significantly higher ACD (t = 4.539, P

Abstract Background To evaluate the feasibility and efficacy of small incision lenticule extraction (SMILE) in the treatment of myopia with corneal opacity. Methods To evaluate the treatment of myopia with corneal opacity, 9 patients (4 males, 5 females) who underwent SMILE were enrolled in this prospective clinical study. One eye of each patient was treated. The results of laser scanning and lenticule extraction were observed during the surgery, and the surgical videos were again reviewed after surgery. Uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), and spherical equivalence (SE) were noted at 1 month after surgery. The depth and density of the corneal opacities were measured by anterior segment optical coherence tomography (AS-OCT) and the Pentacam anterior eye segment analyzer. Results All procedures were uneventful and no intraoperative complications were observed. At 1 month after surgery, the UDVA of all patients was 20/25 or better and no patients lost Snellen lines. The mean safety and efficacy indexes were 1.10 ± 0.24 and 1.08 ± 0.16, respectively, at 1 month postoperatively. The mean postoperative spherical equivalent was 0.27 ± 0.23 diopter (D). All eyes were within ±0.75 D and 8 eyes (88.9%) were within ±0.50 D. There was no eccentric corneal topography or abnormal morphology in the corneal caps. The corneal opacities of all patients were within the optical zone. The mean preoperative depths in the deepest areas of corneal opacity were 152 ± 38 μm (range: 86–217 μm); at 1 month after surgery (P .05), it was 49.8 ± 26.7 (range: 19.8 to 82.5) at 1 month after surgery. Conclusion Patients with corneal opacity can be successfully treated with the SMILE operation. The short-term outcome was good, however the long-term results need further study. Trial registration The trial registration number: ChiCTR-ONRC-13003114 , Date of Last Refreshed on: 2016-01-27, Date of registration on 2013-03-17(retrospectively registered).

Aims To investigate the association between the myopic severity and retinal microvascular density, choroidal vascularity and retrobulbar blood flow in adult anisomyopes. Methods This study comprised 90 eyes of 45 myopic anisomyopes who were recruited for Colour Doppler imaging (CDI) and optical coherence tomography angiography (OCTA). The superficial vessel density (SVD), deep vessel density (DVD), choroidal thickness (ChT) and choroidal vascularity, including total choroidal area (TCA), luminal area (LA), stromal area (SA) and Choroidal Vascularity Index (CVI), were measured using OCTA. Moreover, the Pulsatile Index, peak systolic velocity (PSV) and end diastolic velocity (EDV) of posterior ciliary artery (PCA), central retinal artery (CRA) and ophthalmic artery (OA) were quantified by CDI, and all parameters were compared between two eyes and the correlations among parameters were analysed. Results The mean difference of spherical equivalent (SE) and axial lengths (AL) between eyes were -6.00±2.94 D and 2.48±1.31 mm, respectively. The SVD, DVD, ChT, TCA, LA, SA and CVI were significantly lower in more myopic eyes compared with the contralateral eyes. In more myopic eyes, CDI parameters of CRA and PSV and EDV of PCA were also significantly lower. After adjusting for age and sex, the binocular asymmetry in LA and ChT was independent risk factor affecting interocular difference in both AL and SE. Conclusion Retinal microvascular density, choroidal vascularity and retrobulbar blood flow were simultaneously lower in adult myopic anisomyopes with more myopic eyes and disturbed choroid circulation was related to the severity of myopia. Further longitudinal study was helped to identify the effect of choroidal parameters for myopic progression. [ABSTRACT FROM AUTHOR]

Aims To predict the vault and the EVO-implantable collamer lens (ICL) size by artificial intelligence (AI) and big data analytics. Methods Six thousand two hundred and ninety-seven eyes implanted with an ICL from 3536 patients were included. The vault values were measured by the anterior segment analyzer (Pentacam HR). Permutation importance and Impurity-based feature importance are used to investigate the importance between the vault and input parameters. Regression models and classification models are applied to predict the vault. The ICL size is set as the target of the prediction, and the vault and the other input features are set as the new inputs for the ICL size prediction. Data were collected from 2015 to 2020. Random Forest, Gradient Boosting and XGBoost were demonstrated satisfying accuracy and mean area under the curve (AUC) scores in vault predicting and ICL sizing. Results In the prediction of the vault, the Random Forest has the best results in the regression model (R2=0.315), then follows the Gradient Boosting (R2=0.291) and XGBoost (R2=0.285). The maximum classification accuracy is 0.828 in Random Forest, and the mean AUC is 0.765. The Random Forest predicts the ICL size with an accuracy of 82.2% and the Gradient Boosting and XGBoost, which are also compatible with 81.5% and 81.8% accuracy, respectively. Conclusions Random Forest, Gradient Boosting and XGBoost models are applicable for vault predicting and ICL sizing. AI may assist ophthalmologists in improving ICL surgery safety, designing surgical strategies, and predicting clinical outcomes. [ABSTRACT FROM AUTHOR]

The characterization of variations in riverine microbiota that stem from contaminant sources and transport modes is important for understanding biogeochemical processes. However, the association between complex anthropogenic nitrogen pollution and bacteria has not been extensively investigated owing to the difficulties faced while determining the distribution of nitrogen contaminants in watersheds. Here, we employed the Nitrogen and Oxygen Stable Isotope Tools alongside microbiological analysis to explore microbial characteristics and their responses to complex nitrogen pollution patterns. Significant changes in microbial communities were observed in watersheds with different complex nitrogen pollution sources migrating. Complex nitrogen sources were closely associated with microbiota. The main sources of nitrogen pollution in the dry season watershed were Manurc and Sewage (M&S, 54.2%). M&S exhibited reduced microbial diversity, high number of denitrification groups, and increased nitrogen cycling compared with others. However, Soil Nitrogen sources (SN, 59.3%) in the watershed during the wet season were the main contributors of nitrogen in the watershed. Eluviation allows SN to increased microbial community similarity, in addition to facilitating microbial immobilization processes. Overall, this study expands our knowledge regarding microbial biogeochemistry in catchments and beyond by linking specific nitrogen pollution scenarios to microorganisms.

Abstract How does SARS-CoV-2 cause lung microenvironment disturbance and inflammatory storm is still obscure. We here performed the single-cell transcriptome sequencing from lung, blood, and bone marrow of two dead COVID-19 patients and detected the cellular communication among them. Our results demonstrated that SARS-CoV-2 infection increase the frequency of cellular communication between alveolar type I cells (AT1) or alveolar type II cells (AT2) and myeloid cells triggering immune activation and inflammation microenvironment and then induce the disorder of fibroblasts, club, and ciliated cells, which may cause increased pulmonary fibrosis and mucus accumulation. Further study showed that the increase of T cells in the lungs may be mainly recruited by myeloid cells through ligands/receptors (e.g., ANXA1/FPR1, C5AR1/RPS19, and CCL5/CCR1). Interestingly, we also found that certain ligands/receptors (e.g., ANXA1/FPR1, CD74/COPA, CXCLs/CXCRs, ALOX5/ALOX5AP, CCL5/CCR1) are significantly activated and shared among lungs, blood and bone marrow of COVID-19 patients, implying that the dysregulation of ligands/receptors may lead to immune cell’s activation, migration, and the inflammatory storm in different tissues of COVID-19 patients. Collectively, our study revealed a possible mechanism by which the disorder of cell communication caused by SARS-CoV-2 infection results in the lung inflammatory microenvironment and systemic immune responses across tissues in COVID-19 patients.

Abstract Background Coronavirus disease 2019 (COVID-19) is a respiratory disorder caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which had rapidly spread all over the world and caused public health emergencies in the past two years. Although the diagnosis and treatment for COVID-19 have been well defined, the immune cell characteristics and the key lymphocytes subset alterations in COVID-19 patients have not been thoroughly investigated. Methods The levels of immune cells including T cells, B cells, and natural killer (NK) cells in 548 hospitalized COVID-19 patients, and 30 types of lymphocyte subsets in 125 hospitalized COVID-19 patients admitted to Wuhan Huoshenshan Hospital of China were measured using flow cytometry. The relationship between lymphocytes subsets with the cytokine interleukin-6 (IL-6) and the characteristics of lymphocyte subsets in single-cell RNA sequencing (scRNA-seq) data obtained from peripheral blood mononuclear cells (PBMCs) were also analysed in COVID-19 patients. Results In this study, we found that patients with critical COVID-19 infection exhibited an overall decline in lymphocytes including CD4+ T cells, CD8+ T cells, total T cells, B cells, and NK cells compared to mild and severe patients. However, the number of lymphocyte subsets, such as CD21low CD38low B cells, effector T4 cells, and PD1+ depleted T8 cells, was moderately increased in critical COVID-19 patients compared to mild cases. Notably, except for effector memory T4 cells, plasma blasts and Tregs, the number of all lymphocyte subsets was markedly decreased in COVID-19 patients with IL-6 levels over 30-fold higher than those in healthy cases. Moreover, scRNA-seq data showed obvious differences in the distribution and numbers of lymphocyte subsets between COVID-19 patients and healthy persons, and subsets-specific marker genes of lymphocyte subsets including CD4, CD19, CCR7, and IL7R, were markedly decreased in COVID-19 patients compared with those in healthy cases. Conclusion A comprehensive decrease in immune cell and lymphocyte subsets in critical COVID-19 patients, and peripheral lymphocyte subset alterations showed a clear association with clinical characteristics.

Human babesiosis is an emerging tick-borne disease, caused by haemoprotozoa genus of Babesia. Cases of transfusion-transmitted and naturally acquired Babesia infection have been reported worldwide in recent years and causing a serious public health problem. Babesia duncani is one of the important pathogens of human babesiosis, which seriously endangers human health. The in vitro culture systems of B. duncani have been previously established, and it requires fetal bovine serum (FBS) to support long-term proliferation. However, there are no studies on serum-free in vitro culture of B. duncani. In this study, we reported that B. duncani achieved long-term serum-free culture in VP-SFM AGTTM (VP-SFM) supplemented with AlbuMaxTM I. The effect of adding different dilutions of AlbuMaxTM I to VP-SFM showed that 2 mg/mL AlbuMaxTM I had the best B. duncani growth curve with a maximum percentage of parasitized erythrocytes (PPE) of over 40%, and it can be used for long-term in vitro culture of B. duncani. However, the commonly used 20% serum-supplemented medium only achieves 20% PPE. Clearly, VP-SFM with 2 mg/mL AlbuMaxTM I (VP-SFMA) is more suitable for the in vitro proliferation of B. duncani. VP-SFM supplemented with CD lipid mixture was also tested, and the results showed it could support the parasite growth at 1:100 dilution with the highest PPE of 40%, which is similar to that of 2 mg/mL AlbuMaxTM I. However, the CD lipid mixture was only able to support the in vitro culture of B. duncani for 8 generations, while VP-SFMA could be used for long-term culture. To test the pathogenicity, the VP-SFMA cultured B. duncani was also subjected to hamster infection. Results showed that the hamster developed dyspnea and chills on day 7 with 30% PPE before treatment, which is similar to the symptoms with un-cultured B. duncani. This study develops a unique and reliable basis for further understanding of the physiological mechanisms, growth characteristics, and pathogenesis of babesiosis, and provides good laboratory material for the development of drugs or vaccines for human babesiosis and possibly other parasitic diseases.

Genetic modification provides an invaluable molecular tool to dissect the biology and pathogenesis of pathogens. However, no report is available about the genetic modification of Babesia duncani, a pathogen responsible for human babesiosis that is widespread in North America, suggesting the necessity to develop a genetic manipulation method to improve the strategies for studying and understanding the biology of protozoan pathogens. The establishment of a genetic modification method requires promoters, selectable markers, and reporter genes. Here, the double-copy gene elongation factor-1α (ef-1α) and its promoters were amplified by conventional PCR and confirmed by sequencing. We established a transient transfection system by using the ef-1αB promoter and the reporter gene mCherry and achieved stable transfection through homologous recombination to integrate the selection marker hDHFR-eGFP into the parasite genome. The potential of this genetic modification method was tested by knocking out the thioredoxin peroxidase-1 (TPX-1) gene, and under the drug pressure of 5 nM WR99210, 96.3% of the parasites were observed to express green fluorescence protein (eGFP) by flow cytometry at day 7 post-transfection. Additionally, the clone line of the TPX-1 knockout parasite was successfully obtained by the limiting dilution method. This study provided a transfection method for B. duncani, which may facilitate gene function research and vaccine development of B. duncani.

PURPOSE: To investigate long-term changes in posterior corneal elevation after small incision lenticule extraction (SMILE). METHODS: Thirty-six eyes of 20 patients (7 male, 13 female) who underwent SMILE were recruited for this prospective consecutive study. The mean preoperative manifest spherical equivalent was −6.24 ± 1.47 diopters (D) (range: −3.88 to −8.75 D). Posterior corneal surfaces were measured by a Scheimpflug camera (Pentacam; Oculus Optikgeräte GmbH, Wetzlar, Germany) preoperatively and at 1 month, 6 months, 12 months, 2 years, and 3 years after surgery. Posterior central elevation (PCE), posterior maximal elevation (PME), and posterior elevation at the thinnest corneal point (PTE) in the central 4-mm area above the best-fit sphere (BFS) were analyzed. The BFS was the same across all follow-up periods and determined by the preoperative data. A P value of less than .05 was considered statistically significant. RESULTS: All surgeries were executed without complications and no case of keratectasia was observed during the follow-up. An uncorrected distance visual acuity of 20/20 or better was achieved in 100% of eyes at the last visit. The mean change of PCE, PME, and PTE at 3 years after the operation was −2.39 ± 2.85, 0.50 ± 3.33, and −2.33 ± 2.90 µm, respectively. There were significant differences in the measurements of PCE and PTE before surgery and 3 years after surgery (P ≤ .009); however, there was no significant difference in PME before surgery and 3 years after surgery (P = 1.000). No correlation was found between changes in posterior corneal elevation and residual bed thickness, ablation depth, and preoperative thinnest corneal thickness. CONCLUSIONS: The posterior corneal surface, measured using the Pentacam, was stable after SMILE in the long-term follow-up. The cause of the slight backward change of PCE and PTE needs further study. [ABSTRACT FROM AUTHOR]

A comprehensive understanding of the dynamic changes in interleukin-6 (IL-6) levels is essential for monitoring and treating patients infected with severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2). By analyzing the correlations between IL-6 levels and health conditions, underlying diseases, several key laboratory detection indices, and the prognosis of 1,473 patients with the coronavirus disease 2019 (COVID-19), the role of IL-6 during SARS-CoV-2 infection was demonstrated. Our results indicated that IL-6 levels were closely related to age, sex, body temperature, oxygen saturation (SpO2) of blood, and underlying diseases. As a stable indicator, the changes in IL-6 levels could indicate the inflammatory conditions during a viral infection. Two specific treatments, namely, tocilizumab and convalescent plasma therapy (CPT), decreased the level of IL-6 and relieved inflammation. CPT has an important role in the therapy for patients with critical COVID-19. We also found that patients with IL-6 levels, which were 30-fold higher than the normal level, had a poor prognosis compared to patients with lower levels of IL-6.

Abstract Background Male infertility is a complex disorder caused by genetic, developmental, endocrine, or environmental factors as well as unknown etiology. Polymorphisms in the follicle stimulating hormone beta subunit (FSHB) (rs10835638, c.-211G > T) and follicle stimulating hormone receptor (FSHR) (rs1394205, c.-29G > A; rs6165, c.919A > G; rs6166, c.2039 A > G) genes might disturb normal spermatogenesis and affect male reproductive ability. Methods To further ascertain the aforementioned effects, we conducted a case-control study of 255 infertile men and 340 fertile controls from South China using the Mass ARRAY method, which was analyzed by the t-tests and logistic regression analysis using SPSS for Windows 14.0. In addition, a meta-analysis was performed by combining our results with previous reports using STATA 12.0. Results In the FSHB or FSHR gene single nucleotide polymorphism (SNP) evaluation, no statistically-significant difference was found in the frequency of allelic variants or in genotype distribution between cases and controls. However, a significant association for the comparison of GAA (P: 0.022, OR: 0.63, 95%CI: 0.43–0.94) was seen between the oligozoospermia and controls in haplotype analysis of rs1394205/rs6165/rs6166. In the meta-analysis, rs6165G allele and rs6166 GG genotype were associated with increased risk of the male infertility. Conclusions This study suggested that FSHR GAA haplotype would exert protective effects against male sterility, which indicated that the combination of three SNP genotypes of FSHR was predicted to have a much stronger impact than either one alone. Then in the meta-analysis, a significant association was seen between FSHR rs6165, rs6166 polymorphisms and male infertility. In terms of male infertility with multifactorial etiology, further studies with larger sample sizes and different ethnic backgrounds or other risk factors are warranted to clarify the potential role of FSHB and FSHR polymorphisms in the pathogenesis of male infertility.

African swine fever (ASF), caused by the African swine fever virus (ASFV), is a highly infectious disease afflicting domestic pigs and wild boars. It exhibits an alarming acute infection fatality rate of up to 100%. Regrettably, no commercial vaccines or specific drugs for combating this disease are currently available. This study evaluated the anti-ASFV activities in porcine alveolar macrophages, 3D4/21 cells, and PK-15 cells of four bis-benzylisoquinoline alkaloids (BBAs): cepharanthine (CEP), tetrandrine, fangchinoline, and iso-tetrandrine. Furthermore, we demonstrated that CEP, which exhibited the highest selectivity index (SI = 81.31), alkalized late endosomes/lysosomes, hindered ASFV endosomal transport, disrupted virus uncoating signals, and thereby inhibited ASFV internalization. Additionally, CEP disrupted ASFV DNA synthesis, leading to the inhibition of viral replication. Moreover, berbamine was labeled with NBD to synthesize a fluorescent probe to study the cellular location of these BBAs. By co-staining with Lyso-Tracker and lysosome-associated membrane protein 1, we demonstrated that BBAs target the endolysosomal compartments for the first time. Our data together indicated that BBAs are a class of natural products with significant inhibitory effects against ASFV infection. These findings suggest their potential efficacy as agents for the prevention and control of ASF, offering valuable references for the identification of potential drug targets. IMPORTANCE The urgency and severity of African swine fever (ASF) underscore the critical need for effective interventions against this highly infectious disease, which poses a grave threat to domestic pigs and wild boars. Our study reveals the potent anti-African swine fever virus (ASFV) efficacy of bis-benzylisoquinoline alkaloids (BBAs), particularly evident in the absence of progeny virus production under a 5 µM concentration treatment. The structural similarity among cepharanthine, tetrandrine, fangchinoline, and iso-tetrandrine, coupled with their analogous inhibitory stages and comparable selectivity indexes, strongly suggests a shared antiviral mechanism within this drug category. Further investigation revealed that BBAs localize to lysosomes and inhibit the internalization and replication of ASFV by disrupting the endosomal/lysosomal function. These collective results have profound implications for ASF prevention and control, suggesting the potential of the investigated agents as prophylactic and therapeutic measures. Furthermore, our study offers crucial insights into identifying drug targets and laying the groundwork for innovative interventions. [ABSTRACT FROM AUTHOR]