Whole-genome sequencing of Paracoccus species isolated from the healthy human eye and description of Paracoccus shanxieyensis sp. nov.

Currently, the genus Paracoccus comprises 76 recognized species. Members of Paracoccus are mostly isolated from environmental, animal, and plant sources. This report describes and proposes a novel species of Paracoccus isolated from clinical specimens of the human ocular surface. We isolated two aerobic, Gram-stain-negative, non-spore-forming, coccoid or short rod-shaped, and non-motile strains (designated DK398T and DK608) from conjunctival sac swabs of two healthy volunteers. The results showed that the strains grew best under the conditions of 28°C, pH 7.0, and 1.0 % (w/v) NaCl. Sequence analysis based on the 16S rRNA gene showed that strains DK398T and DK608 were members of Paracoccus, most similar to Paracoccus laeviglucosivorans 43PT (98.54 and 98.62 %), Paracoccus litorisediminis GHD-05T (98.34 and 98.41 %), and Paracoccus limmosus NB88T (98.21 and 98.29 %). Phenotypic analysis showed that DK398T and DK608 were positive for catalase and oxidase, negative for producing N-acetyl-ß-glucosaminic acid, arginine dihydrolase, and ß-glucuronidase but positive for leucine arylamidase. The predominant isoprenoid quinone was Q-10, and the major polar lipids included phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, and an unidentified glycolipid. The major fatty acids (>10%) were summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and C16 : 0. The meso-diaminopimelic acid was found in the cell wall peptidoglycan of DK398T. The major cell wall sugars were ribose and galactose. Based on the results of phylogenetic analyses, low (<83.22 %) average nucleotide identity, digital DNA-DNA hybridization (<26.0%), chemotaxonomic analysis, and physiological properties, strain DK398T represents a novel species of the genus Paracoccus, for which the name Paracoccus shanxieyensis sp. nov. is proposed. The type strain is DK398T (=CGMCC 1.17227T=JCM 33719T).

Prognosis and Prediction of Asymptomatic Intracranial Hemorrhage After Endovascular Thrombectomy: A Multi-Center Study.

PURPOSE: The impact of asymptomatic intracranial hemorrhage (aICH) on functional outcomes after endovascular thrombectomy (EVT) remains unclear, and tools for forecasting this complication are lacking. We aim to evaluate the clinical relevance of aICH and establish a prediction model. METHODS: Data of patients who received EVT for acute anterior-circulation large vessel occlusion in 3 comprehensive hospitals were retrospectively analyzed. Asymptomatic intracranial hemorrhage was defined as any hemorrhage detected after EVT that did not fulfill the definition of symptomatic intracranial hemorrhage in the European Cooperative Acute Stroke Study. Logistic regression models were performed to assess the impact of aICH on 90-day functional outcomes and identify the predictors of aICH, which were then used to establish a prediction model. The discrimination, calibration, and clinical utility of the model were evaluated. RESULTS: This study included 460 patients, among whom 152 (33.0%) developed aICH after EVT. Asymptomatic intracranial hemorrhage was negatively associated with 90-day excellent outcomes (adjusted odds ratio [OR]: 0.414, 95% confidence interval [CI]: 0.230-0.745, p=0.003) and good outcome (adjusted OR: 0.603, 95% CI: 0.374-0.971, p=0.037), but not with mortality (adjusted OR: 1.110, 95% CI: 0.611-2.017, p=0.732) after adjusted for other predictors of functional outcome. Pre-stroke anticoagulant therapy (OR: 2.233, 95% CI: 1.073-4.647, p=0.032), Alberta stroke program early CT score (OR: 0.842, 95% CI: 0.754-0.939, p=0.002), site of occlusion (internal carotid artery occlusion as the reference; M1 segment of middle cerebral artery occlusion, OR: 2.827, 95% CI: 1.409-5.674, p=0.003; tandem occlusion, OR: 3.928, 95% CI: 1.752-8.806, p=0.001), intravenous thrombolysis (OR: 2.091, 95% CI: 1.362-3.209, p=0.001), and successful recanalization (OR: 0.383, 95% CI: 0.213-0.689, p=0.001) were identified as the predictors of aICH, which were incorporated into a nomogram model. The area under the receiver operating characteristic curve of the model was 0.707 (95% CI: 0.657-0.757), and the calibration plot demonstrated good consistency between actual observed and predicted probability of aICH. Decision curve analysis showed that patients might benefit from the model. CONCLUSION: Asymptomatic intracranial hemorrhage was negatively associated with favorable functional outcome after EVT. We established a nomogram model for predicting aICH, which requires external clinical validation. CLINICAL IMPACT: The impact of asymptomatic intracranial hemorrhage after endovascular thrombectomy on mid-term functional outcome has been controversial. We found that asymptomatic intracranial hemorrhage may also decreased the likelihood of 90-day favourable functional outcome after endovascular thrombectomy, supporting the notion that asymptomatic intracranial hemorrhage at the acute stage may not be benign. Moreover, we established a prediction model for this complication, which may improve clinical evaluation and management of patients who would receive endovascular thrombectomy for large vessel occlusion.

A combinatorial strategy for overcoming primary and acquired resistance of MEK inhibition in colorectal cancer.

Compared with traditional chemotherapeutic drugs, targeted therapeutic medicine has the advantages of high efficacy and less toxic side effects. However, in clinical practice for treatment of colorectal cancer, the primary and acquired resistance of these medicines limits their effectiveness in targeted therapy, therefore impedes the development of precision medicine and personalized therapy. Currently, there are limited number of drugs for targeted therapy of colorectal cancer, mainly monoclonal antibodies against EGFR or VEGFR inhibitors. Trametinib, a MEK inhibitor, has been applied in melanoma patient successfully, but not been used in clinical treatment of colorectal cancer because of its drug resistance. To identify the resistance mechanism of colorectal cancer cells to trametinib and find useful chemical combination to overcome the resistance, we screened primary and acquired cell line first and then tested multiple synergistic drug combinations by using the Chou-Talalay method. We obtained the primary resistant cell lines SW480, CW-2 and the acquired drug-resistant cell line RKO-R as well as a synergistic combination of trametinib and GSK2126458. This combination inhibits the colony formation of colorectal cancer cells and the growth of xenograft tumors in nude mice. Mechanistic analysis showed that trametinib can activate the alternative PI3K-AKT signaling pathway while inhibiting the MAPK pathway, which may be one of the molecular mechanisms of primary and acquired trametinib tolerance in colorectal cancer cells. Importantly, this bypass activation can be blocked by GSK2126458. These results suggest that a combination of trametinib and GSK2126458 is an effective approach for treating colorectal cancer resistance to trametinib.

MRPL35 Is Up-Regulated in Colorectal Cancer and Regulates Colorectal Cancer Cell Growth and Apoptosis.

Mitochondrial ribosome proteins (MRPs), which are encoded by the nuclear genomic DNA, are important for mitochondrial-encoded protein synthesis and mitochondrial function. Emerging evidence suggests that several MRPs also exhibit important extra-mitochondrial functions, such as involvement in apoptosis, protein biosynthesis, and signal transduction. In this study, we demonstrate a significant role of MRP L35 (MRPL35) in colorectal cancer (CRC). The expression of MRPL35 was higher in CRC tissues than in matched cancer-adjacent tissues and higher in CRC cells than in normal mucosal epithelial cells. Higher MRPL35 expression in CRC tissue correlated with shorter overall survival for CRC patients. In vitro, down-regulation of MRPL35 led to increased production of reactive oxygen species (ROS) together with DNA damage, loss of cell proliferation, G2/M arrest, a decrease in mitochondrial membrane potential, apoptosis, and autophagy induction. MRPL35 knockdown inhibited tumor proliferation in a CRC xenograft nude mouse model. Furthermore, overexpression of MRPL35 or treatment of cells with the ROS scavenger, N-acetyl cysteine, abrogated ROS production, cell cycle arrest, and apoptosis in vitro. These findings suggest that MRPL35 plays an essential role in the development of CRC and may be a potential therapeutic target for CRC.

Yield advantage and cadmium decreasing of rice in intercropping with water spinach under moisture management.

Rice (Oryza sativa L.) intercropping with water spinach (Ipomoea aquatica Forsk) is an effective agricultural practice for safe crop production and for phytoremediation in cadmium-contaminated soil. A field and pot experiment were conducted to investigate the growth and cadmium absorption of rice intercropped with water spinach under different moisture management schemes (continuous flooding, interval flooding, and 75% field capacity). In the field experiment, the concentration of Cd in the grain of rice was significantly lower in the intercropping system than that permitted by the National Food Safety Standard of China (GB 2762-2017). Furthermore, the land equivalent ratio (1.42) was higher in the rice-water spinach intercropping system, indicating a significant advantage of the intercropping system in yield. At the same time, the bio-concentration amount (BCA) of Cd of rice and water spinach in intercropping system significantly increased by 17.99% and 31.98%, respectively (P＜0.05). However, the metal removal equivalent ratio (MRER) of Cd was 1.34, which showed the intercropping system of rice-water spinach had advantage in Cd removal. In the pot experiment, the total iron plaque concentration on the root surface of rice and the pH of the rhizosphere soil were higher under continuous flooding (TCF) than under the control conditions (75% field capacity, TCK), which could significantly decrease the available Cd in the rhizosphere soil and the accumulation of Cd in rice organs. So, this study demonstrated that iron plaque can obstruct and decrease the Cd absorbed by rice in a rice-water spinach intercropping system combined with water management. The intercropping rice with water spinach can achieve the goal of remediation while producing for farmland contaminated by Cd.

Difference between arterial and venous peak optical density after thrombectomy is associated with functional outcomes.

Background: The density of contrast medium in digital subtraction angiography (DSA) have been used to evaluate the cerebral circulation function. Our aim was to study the effect of difference in arteriovenous peak optical density (POD) after thrombectomy on functional outcomes. Methods: Consecutive patients with acute ischemic stroke due to large vessel occlusion who underwent thrombectomy were reviewed. We processed DSA images with ImageJ software to measure the POD of internal carotid artery (ICA) and cortical veins. The average POD of cortical veins (PODVA) and the POD difference between ICA and cortical veins (PODICA-CV) were calculated. Primary outcome was good functional outcome (modified Rankin scale score of 0-2 at 90 days). Results: One hundred sixty-six patients were finally included in the study. Patients with good functional outcome had lower ipsilateral PODVA (median [interquartile range (IQR)], 257.198 [216.623-296.631] vs. 290.944 [248.647-338.819], p < 0.001) and lower ipsilateral PODICA-CV (median [IQR], 128.463 [110.233-153.624] vs. 182.01 [146.621-211.331], p < 0.001). Multivariable logistic regression analyses showed that ipsilateral PODVA (odds ratio [OR] 0.991, 95% confidence interval [CI] 0.984-0.999, p = 0.019) and ipsilateral PODICA-CV (OR 0.975, 95% CI 0.963-0.986, p < 0.001) were associated with good functional outcome. The predictive ability was significantly enhanced in the model including ipsilateral PODICA-CV (0.893 vs. 0.842, p = 0.027). No correlation was found between ipsilateral PODICA-CV and expanded Thrombolysis in Cerebral Infarction grades (r = -0.133, p = 0.099). Conclusion: Ipsilateral PODICA-CV is an additional indicator of cerebral reperfusion status and predicts functional outcomes after thrombectomy.

Case report: Retrograde endovascular recanalization of vertebral artery occlusion with non-tapered stump via the deep cervical collateral.

Introduction: Vertebral artery (VA) occlusive disease is the major cause of posterior circulation ischemic stroke. Endovascular recanalization has been reported as a feasible treatment for patients with symptomatic VA occlusion refractory to optimal medical therapy. However, VA occlusion with non-tapered stump exhibits a low technique success rate when treated by antegrade endovascular therapy because of increased difficulty in passing the guidewire into the occluded segment. Herein, we presented a novel endovascular approach to recanalize chronically occluded VA with a non-tapered stump using a retrograde method via the deep cervical collateral, which has not been reported before. Case presentation: The present case was a patient with VA ostial occlusion with non-tapered stump and distal severe stenosis of the left VA who had recurrent posterior circulation transit ischemic attacks under optimal medical therapy. CT angiography demonstrated proximal non-tapered occlusion and distal severe stenosis of the left VA, and that the right VA did not converge with the left VA into basilar artery. Endovascular treatment was recommended and performed on this patient. However, antegrade endovascular recanalization of the left VA origin occlusion failed because the micro guidewire was unable to traverse the occluded segment. Fortunately, robust collateral from the deep cervical artery to the V3 segment of the left VA developed, in which we advanced the micro guidewire to the V3 segment of the left VA and reversely passed the micro guidewire through the occluded segment. Then, the occlusion and stenosis of the left VA were successfully resolved with angioplasty and stenting. After the procedure, the patient reported no neurological symptoms under medical therapy during 3-month follow-up. Conclusion: Antegrade endovascular recanalization of VA occlusion with a non-tapered stump is a challenge. The retrograde endovascular method via the cervical collateral may be an alternative for this type of VA occlusion, which requires further exploration.

Proximal hyperdense middle cerebral artery sign is associated with increased risk of asymptomatic hemorrhagic transformation after endovascular thrombectomy: a multicenter retrospective study.

OBJECTIVE: To investigate whether hyperdense middle cerebral artery sign (HMCAS) on pretreatment no-contrast CT (NCCT) is associated with hemorrhagic transformation (HT) after endovascular thrombectomy (EVT). METHODS: Patients with acute middle cerebral artery (MCA) occlusion who received EVT in three comprehensive hospitals were retrospectively evaluated. They were divided into four groups based on the presence or absence of HMCAS and corresponding CTA findings, among whom differences were compared. Univariate and multivariate logistic regression analyses were performed to investigate the association between HMCAS and HT and its subtypes. RESULTS: 318 patients were included, among whom 149 (46.9%) had HMCAS. Patients in the proximal positive HMCAS group had higher National Institute of Health Stroke Scale scores and lower Alberta Stroke Program Early CT Scores (ASPECTS) than those in the proximal negative HMCAS group. The rate of HT was higher in the proximal positive HMCAS group than that in the proximal negative HMCAS group. In multivariate logistic regression analysis, the proximal HMCAS were independently associated with HT (adjusted OR = 2.073, 95% CI 1.211-3.551, p = 0.008) and aHT (adjusted OR = 2.271, 95% CI 1.294-3.986, p = 0.004), but not with sHT. Patients who developed HT, including aHT and sHT, had a lower rate of good outcome. CONCLUSION: Proximal HMCAS on initial NCCT was independently associated with aHT in patients who received EVT for acute MCA occlusion. Both aHT and sHT had a detrimental effect on clinical outcome.

Characteristics and associations of microbial community in the eye, anterior nares, and oropharynx of healthy adults: eye-nose microbiota transmission.

PURPOSE: We compared the microbial communities of the eye, anterior nares (ANs) and oropharynx (OP) of healthy adults to provide a basic understanding of the microbial associations among the three sites. METHODS: The name of the registry of our prospective study was "Study on the diversity of microbial flora in the eye, nose and oropharynx of healthy people". The trial number is ChiCTR2300067724 (https://www.chictr.org.cn/index.aspx). Swabs were collected from the eye, ANs and OP of 48 healthy adult participants for 16S rRNA gene amplicon sequencing. The bacterial community profiles and their functional associations were compared among the three sites. RESULTS: At a phylum level, the basic bacterial compositions in the eye and ANs were generally similar, and the predominant phyla were Actinobacteria, Proteobacteria and Firmicutes. In contrast, the OP microbiota was characterized by an increased abundance of Bacteroidetes. At a genus level, Corynebacterium, Cutibacterium and Staphylococcus were the most abundant in the eye and ANs. Prevotella-7, Alloprevatella, Haemophilus and Streptococcus were more abundant in the OP. Correlation analysis of the eye and ANs microbiota suggested that Cutibacterium and Micrococcus may migrate from the eye to nose (P < 0.05). CONCLUSIONS: The bacterial flora composition and function predictions of eye and ANs were similar, but differed from those of the OP in healthy adults. The OP bacterial flora distribution was markedly different, showing characteristics similar to that of the digestive tract flora. Thus, the eye and ANs microorganisms may be related in healthy individuals. Cutibacterium and Micrococcus may migrate from the eye to the nose.

Effects of different planting distances and fertilizer use on the remediation of farmland contaminated with Cd by intercropping Cucurbita moschata and Amaranthus hypochondriacus L.

Selecting suitable agronomic measures can strengthen the application of intercropping in the remediation of cadmium (Cd)-contaminated soil. In this study, the effects of different planting densities and fertilizer applications on the crop growth and Cd absorption of a pumpkin (Cucurbita moschata)-Amaranthus hypochondriacus L. intercropping system was determined. The goal was to provide enhanced means and a scientific basis for the promotion and application of this intercropping system. The Cd content of pumpkin in different planting systems was lower than the national food safety standard (0.05 mg kg-1). In the IN-1 (4 pumpkin plants intercropped with 200 A. hypochondriacus plants) and IN-2 (4 pumpkin plants intercropped with 400 A. hypochondriacus plants) intercropping systems, the bioconcentration amount (BCA) per plant of Cd in A. hypochondriacus increased by 32.43% and 25.25%, respectively, compared with that of the monocropping system (P < 0.05). The IN-2 system had the highest equivalent ratio of heavy metal removal (3.08), indicating that this model had a substantial advantage for removing Cd. The land equivalent ratio of IN-1 (2.89) and IN-2 (2.60) was significantly higher than that of other intercropping systems, indicating that these two models had obvious yield advantages. In our study, chicken manure was the best at promoting the growth and yield of the two plants and sludge treatment significantly enhance Cd absorption of A. hypochondriacus. In general, intercropping four pumpkin plants with 400 A. hypochondriacus plants and applying chicken manure fertilizer can strengthen the application of this intercropping system in Cd-contaminated soil.

[Impact of Meteorological Conditions on PM2.5 in Jiangsu Province from 2001 to 2019].

The weather research and forecasting model coupled with chemistry (WRF-Chem) was used to investigate the impact of meteorological conditions on PM2.5 in Jiangsu Province from 2001 to 2019. Under the condition of constant emission sources, the strongest positive and negative anomalies of annual PM2.5 concentration caused by meteorological conditions occurred in 2008 and 2001, respectively. Furthermore, their anomalies respectively accounted for 10.5% and -14.3% relative to the long-time averaged annual PM2.5 concentration, indicating that meteorological conditions are an important factor causing the interannual variation in PM2.5 concentration in Jiangsu Province. The empirical orthogonal function decomposition results show that the influence of meteorological conditions on the spatial distribution of PM2.5 concentration in Jiangsu Province is consistent under this mode. Additionally, the boundary layer height, temperature, relative humidity, wind speed, and precipitation all have significant negative correlations with the PM2.5 concentration in Jiangsu Province. Meanwhile, the linear regression equation constructed by the above meteorological factors can characterize the relationship between PM2.5 concentration and meteorological conditions well. Moreover, the correlation between the fitting value and the simulated value was 0.73, which was statistically significant at a 99% confidence level according to a student's t test.

A Nomogram for Predicting Symptomatic Intracranial Hemorrhage after Endovascular Thrombectomy.

BACKGROUND: Symptomatic intracranial hemorrhage (sICH) is a devastating complication of endovascular thrombectomy (EVT). We aim to develop and validate a nomogram for predicting sICH in patients with large vessel occlusion (LVO) in the anterior circulation. METHODS: We performed a single-center retrospective analysis on collected data from patients undergoing EVT for LVO in the anterior circulation between January 2018 and December 2021. Forward stepwise logistic regression was performed to identify independent predictors of sICH and establish a nomogram. The discrimination and calibration of the model was accessed using the area under the receiver operating characteristic curve (AUC-ROC) and calibration plot. The model was internally validated using bootstrap and 5-fold cross-validation. RESULTS: 243 patients were included, among whom 23 developed sICH (9.5%). After multivariate logistic regression, baseline glucose level (odds ratio [OR], 1.16; p = 0.022), Alberta Stroke Program Early CT Score (OR, 0.44; p < 0.001), regional Leptomeningeal Collateral score (OR, 0.74; p < 0.001) were identified as independent predictors of sICH, which were then incorporated into a predictive nomogram. The ROC curve of the model showed good discriminative ability with an AUC of 0.856 (95% CI: 0.785-0.928). The calibration plot of the model demonstrated good consistency between the actual observed and the predicted probability of sICH. The model was internally validated by using bootstrap (1000 resamples) with an AUC of 0.835 (95%CI: 0.782-0.887) and 5-fold cross-validation with an AUC of 0.831 (95%CI: 0.775-0.887). CONCLUSION: Our model is a reliable tool to predict sICH after EVT. Although the model was internally validated, further external validation is also warranted.

Posterior Circulation ASPECTS on CT Angiography Predicts Futile Recanalization of Endovascular Thrombectomy for Acute Basilar Artery Occlusion.

Background: Acute basilar artery occlusion (BAO) is the most potentially disastrous outcome and has a high risk of recurrence stroke in posterior circulation infarction (PCI). However, the rate of futile recanalization remains high despite successful recanalization. The objective of this study was to investigate 90 days functional outcomes among patients with BAO who underwent endovascular thrombectomy (EVT) and to identify the risk factors associated with futile recanalization. Methods: We retrospectively analyzed 72 patients with acute BAO who received EVT from January 2018 to June 2021. CT angiography source images posterior circulation Acute Stroke Prognosis Early CT Score (CTA-SI pc-ASPECTS) evaluated the extensive hypoattenuation in patients with BAO. Futile recanalization defined an modified Rankin Scale (mRS) of 3-6 at 90 days despite a successful recanalization. Logistic regression analysis was performed to investigate the predictors of futile recanalization. Results: Our sample included a total of 55 eligible patients. Patients with poor outcomes showed that the pc-ASPECTS score was lower in patients with poor outcomes than that in patients with good outcomes (P = 0.017). Longer time from symptoms onset-to-the puncture (P = 0.014) and elevation of leucocytes (P = 0.012) were associated with poor outcomes. The multivariable logistic analysis showed that pc-ASPECTS and onset-to-puncture time (OPT) were independent predictors of futile recanalization. Conclusions: This study suggested that pc-ASPECTS and OPT are independent predictors of futile recanalization after EVT in patients with BAO. The lower pc-ASPECTS score and longer puncture time will have a poor clinical outcome.

Clot Meniscus Sign Is Associated With Thrombus Permeability and Choice of Mechanical Thrombectomy Technique in Acute Middle Cerebral Artery Occlusion.

Background and Purpose: The method of mechanical thrombectomy (MT) is related to vascular anatomy and stroke etiology. Meniscus sign and thrombus permeability as imaging markers may be instructive for the selection of MT. This study aims to clarify the relationship among meniscus sign, thrombus permeability, and choice of MT in patients with acute middle cerebral artery occlusion. Materials and Methods: A total of 111 patients with acute middle cerebral artery occlusion (MCAO) who underwent MT were retrospectively analyzed. Clot meniscus sign was defined as the appearance of meniscoid/edge-like or single- or double-wall contrast channels besides or around insular blood clots. The radiographic, clinical, and surgical data of patients with MCAO with or without meniscus sign were compared. Results: The meniscus sign positive group (n = 26) has higher thrombus permeability (HUs) (26.92 ± 9.69 vs. 22.84 ± 7.88, p = 0.031) than those without it. Shorter puncture-to-recanalization (P2R) time (65.5 vs. 88, p = 0.012), higher complete recanalization rate (85.71 vs. 33.33%, p < 0.01), and better clinical outcome (p < 0.01) were obtained by selecting contact aspiration (CA) over stent retriever (SR) in patients with positive meniscus sign. In patients with negative meniscus sign, there was no significant difference in clinical outcome after receiving CA or SR. Conclusion: Patients with MCAO with positive meniscus sign have higher thrombus permeability and are more suitable for CA to acquire better clinical outcomes.

Interplanting of rice cultivars with high and low Cd accumulation can achieve the goal of "repairing while producing" in Cd-contaminated soil.

Interplanting has been highlighted as a promising, cost-effective, and environmentally friendly solution for the remediation of contaminated soil. In this study, field experiments were conducted to study growth and cadmium (Cd) uptake in monoculture and interplanting systems with rice varieties Changliangyou 772 (C-772) and Changxianggu (Cho-ko-koku). And a pot culture experiment was conducted to investigate the response of the rhizosphere microecology of these two rice varieties. In the interplanting system of the field study, the Cd concentration in the grains of C-772 was significantly reduced (P < 0.05) from 0.30 mg kg-1 to 0.16 mg kg-1 and reached the national food safety standard of China (GB2762-2017, 0.20 mg kg-1), whereas the bioconcentration amount (BCA) per plant of Cd in Cho-ko-koku was significantly increased by 68.18 % (P < 0.05). The land equivalent ratio (LER) and Cd metal removal equivalent ratio (MRER) of the interplanting system were 1.03 and 1.05, illustrating that the interplanting system was superior in terms of yield and Cd elimination. In the pot experiment, the interplanting system significantly (P < 0.05) reduced the iron plaque content on the root surface and organic acids content in the rhizosphere environment of C-772 while markedly increasing those levels in Cho-ko-koku. At ripening stage, the interplanting system significantly decreased the rhizosphere available Cd concentration of C-772 from 0.38 mg kg-1 to 0.22 mg kg-1 (P < 0.05), while significantly increased the rhizosphere available Cd concentration of Cho-ko-koku from 0.27 mg kg-1 to 0.32 mg kg-1 (P < 0.05). Thus, Cd uptake of C-772 and Cho-ko-koku showed apparent differences. Oxalic and tartaric acid were identified as the most crucial factors affecting Cd uptake by C-772 and Cho-ko-koku in the interplanting system, respectively. In summary, this interplanting system is a promising planting pattern that can simultaneously improve land use efficiency and alleviate Cd pollution.

Application of High-Throughput Sequencing Technology in Identifying the Pathogens in Endophthalmitis.

Infectious endophthalmitis is an important cause of vision loss worldwide. It is an inflammatory reaction caused by bacteria, fungi, and other micro-organisms and often occurs as a complication of intraocular surgery, especially following cataract surgery or intravitreal injection. The focus of the prevention and treatment of infectious endophthalmitis is the early detection of microbial flora, such as fungi or bacteria. Current identification methods for bacteria include Gram staining-based, culture-based, and polymerase chain reaction (PCR)-based methods. The matrix-assisted laser desorption/ionization time-of-flight mass spectrometry technology is now the standard identification method of bacteria and fungi after their isolation in culture. The remarkable sensitivity of PCR technology for the direct detection of micro-organisms in clinical samples makes it particularly useful in culture-positive and culture-negative endophthalmitis. Furthermore, PCR increases the rate of microorganism detection in intraocular samples by 20% and can provide a microbiology diagnosis in approximately 44.7-100% of the culture-negative cases. This review aims to introduce the development of different methods for the detection and identification of micro-organisms causing endophthalmitis through a literature review; introduce the research status of the first, second, and third-generation sequencing technologies in infectious endophthalmitis; and understand the research status of endophthalmitis microbial flora. For slow-growing and rare micro-organisms, high-throughput sequencing (HTS) offers advantages over conventional methods and provides a basis for the identification of pathogens in endophthalmitis cases with negative culture. It is a reliable platform for the identification of pathogenic bacteria of infectious endophthalmitis in the future and provides a reference for the clinical diagnosis and treatment of infectious endophthalmitis. The application of HTS technology may also be transformative for clinical microbiology and represents an exciting future direction for the epidemiology of ocular infections.

Comparison of the ocular surface microbiota between thyroid-associated ophthalmopathy patients and healthy subjects.

Purpose: Thyroid-associated ophthalmopathy (TAO) is a chronic autoimmune disease. In this study, high-throughput sequencing was used to investigate the diversity and composition of the ocular microbiota in patients with TAO. Methods: Patients with TAO did not receive treatment for the disease and did not have exposed keratitis. Patients with TAO (TAO group) and healthy individuals (control group) were compared. All samples were swabbed at the conjunctival vault of the lower eyelid. The V3 to V4 region of the 16S rDNA was amplified using polymerase chain reaction and sequenced on the Illumina HiSeq 2500 Sequencing Platform. Statistical analysis was performed to analyze the differences between the groups and the correlation between ocular surface microbiota and the disease. The ocular surface microbiota of patients and healthy individuals were cultured. Results: The ocular surface microbiota structure of TAO patients changed significantly. The average relative abundance of Bacillus and Brevundimonas increased significantly in the TAO group. Corynebacterium had a significantly decreased relative abundance (P<0.05). Paracoccus, Haemophilus, Lactobacillus, and Bifidobacterium were positively correlated with the severity of clinical manifestations or disease activity (P<0.05). Bacillus cereus and other opportunistic pathogens were obtained by culture from TAO patients. Conclusions: This study found that the composition of ocular microbiota in patients with TAO was significantly different from that in healthy individuals. The ocular surface opportunistic pathogens, such as Bacillus, Brevundimonas, Paracoccus, and Haemophilus in TAO patients, increase the potential risk of ocular surface infection. The findings of this study provide a new avenue of research into the mechanism of ocular surface in TAO patients.

The species-level microbiota of healthy eyes revealed by the integration of metataxonomics with culturomics and genome analysis.

Objectives: To characterize the healthy ocular surface microbiota at the species level, including cultured and uncultured taxa. Methods: We integrated the metataxonomic method with culturomics and genome sequencing analysis of selected isolated strains to better illustrate the taxonomic structure of the ocular surface microbiota. The metataxonomics used the full-length 16S rRNA gene sequences and the operational phylogenetic unit strategy, which can precisely identify the cultured and uncultured or potentially new taxa to species level based on the phylogenetic tree constructed. Results: We detected 1,731 operational phylogenetic units (OPUs) in 196 healthy eyes from 128 people, affiliated to 796 cultured species, 784 potentially new species, and 151 potentially new higher taxa. The microbiota for each eye had 49.17 ± 35.66 OPUs. Of the 796 cultured species, 170 (21.36%) had previously caused clinical infections. Based on where they were initially isolated, the ocular surface microbiota mainly came from human body sites (34.55%), the environment (36.93%), plants (9.05%), animals (4.90%), and others; 428 strains were isolated from 20 eyes, affiliated to 42 species, and had come from the environment (33.33%) and the skin (16.67%). Of these, 47.62% had previously caused clinical infections. Genome analysis of 73 isolators revealed that 68.5% of them carried antibiotic resistance genes. The most frequently isolated genera, namely Staphylococcus, Streptococcus, and Moraxella, had an average of 5.30, four, and three resistance genes per strain, respectively. Discussion: The study found that the ocular surface microbiota mainly came from the environment, plants, animals, food, and human body sites such as the skin, oral cavity, upper respiratory tract, etc. No core member of ocular surface microbiota was detected at the species level. The human eyes were invaded and colonized by bacteria from the exposed environment, some of which were capable of causing infections in humans and carried antibiotic resistance genes. Preventive measures should be developed to protect our eyes from danger.

A Loop-mediated Isothermal Amplification With a Nanoparticle-Based Lateral Flow Biosensor Assay to Detect Pseudomonas aeruginosa in Endophthalmitis.

Purpose: Pseudomonas aeruginosa is the most common bacteria causing endophthalmitis after cataract surgery. Vitreous fluid culture and molecular studies are commonly used in clinical diagnoses, but have disadvantages, such as a long culture cycle and low detection sensitivity. Here, we report a loop-mediated isothermal amplification (LAMP) method combined with the nanoparticles-lateral flow biosensor (LFB) method for rapid and specific detection of P. aeruginosa. Methods: A set of six primers was designed to target the OprL gene of P. aeruginosa. Genomic DNA extracted from several gram-negative and gram-positive bacteria was used to determine the sensitivity and specificity of the analysis. LAMP reactions were conducted at 65 °C for 50 minutes, and results were reported using the LFB method. Results: The DNA template of P. aeruginosa was specifically recognized by the P. aeruginosa-LAMP-LFB (PA-LAMP-LFB) method as no cross reactions were observed for non-P. aeruginosa templates. The analytical sensitivity of our assay was 100 fg per test for the pure cultured DNA template, and the result obtained using the LFB was consistent with that of colorimetric indicator detection. The whole test could be completed within 1h. This method was used to detect P. aeruginosa, Staphylococcus aureus, and Klebsiella pneumoniae; only P. aeruginosa was positive. The positive rates of P. aeruginosa detected by a traditional culture method, the LAMP-LFB method, and the fluorescence quantitative polymerase chain reaction method were 17.7%, 17.7%, and 13.3%, respectively. Conclusions: The P. aeruginosa-LAMP-LFB method established here is a rapid, specific, and sensitive method for the detection of P. aeruginosa, which can be widely used.