Constructing 2D Phthalocyanine Covalent Organic Framework with Enhanced Stability and Conductivity via Interlayer Hydrogen Bonding as Electrocatalyst for CO2 Reduction.

Fabricating COFs-based electrocatalysts with high stability and conductivity still remains a great challenge. Herein, 2D polyimide-linked phthalocyanine COF (denoted as NiPc-OH-COF) is constructed via solvothermal reaction between tetraanhydrides of 2,3,9,10,16,17,23,24-octacarboxyphthalocyaninato nickel(II) and 2,5-diamino-1,4-benzenediol (DB) with other two analogous 2D COFs (denoted as NiPc-OMe-COF and NiPc-H-COF) synthesized for reference. In comparison with NiPc-OMe-COF and NiPc-H-COF, NiPc-OH-COF exhibits enhanced stability, particularly in strong NaOH solvent and high conductivity of 1.5 × 10-3 S m-1 due to the incorporation of additional strong interlayer hydrogen bonding interaction between the O-H of DB and the hydroxy "O" atom of DB in adjacent layers. This in turn endows the NiPc-OH-COF electrode with ultrahigh CO2-to-CO faradaic efficiency (almost 100%) in a wide potential range from -0.7 to -1.1 V versus reversible hydrogen electrode (RHE), a large partial CO current density of -39.2 mA cm-2 at -1.1 V versus RHE, and high turnover number as well as turnover frequency, amounting to 45 000 and 0.76 S-1 at -0.80 V versus RHE during 12 h lasting measurement.

Antidepressant polyprenylated acylphloroglucinols from Hypericum ascyron.

Phytochemical investigation on the 90% EtOH extract of the air-dried aerial parts of Hypericum ascyron resulted in the isolation of three new polycyclic polyprenylated derivatives ascyronines A-C (1-3). Structural elucidation of all the compounds was performed by spectral methods such as 1D and 2D (1H-1H COSY, HMQC, and HMBC) NMR spectroscopy. All the polycyclic polyprenylated acylphloroglucinols were evaluated for their antidepressant activity by inhibiting the reuptake of tritiated serotonin ([3H]-5-HT) and noradrenalinet ([3H]-NE) in rat brain synaptosomes. Compounds 2 and 3 exhibited weak antidepressant activities in the [3H]-5-HT mode.

Semisolid Extrusion 3D Printing of Propranolol Hydrochloride Gummy Chewable Tablets: an Innovative Approach to Prepare Personalized Medicine for Pediatrics.

The demand for personalized medicine has received extensive attention, especially in pediatric preparations. An emerging technology, extrusion-based 3D printing, is highly attractive in the field of personalized medicine. In this study, we prepared propranolol hydrochloride (PR) gummy chewable tablets tailored for children by semisolid extrusion (SSE) 3D printing technology to meet personalized medicine needs in pediatrics. In this study, the effects of critical formulation variables on the rheological properties and printability of gum materials were investigated by constructing a full-factorial design. In addition, the masticatory properties, thermal stability, and disintegration time of the preparations were evaluated. Bitterness inhibitors were used to mask the bitterness of the preparations. The results of the full-factorial design showed that the amount of gelatin and carrageenan were the key factors in the formulation. Gelatin can improve printability and masticatory properties, carrageenan can improve thermal stability, and accelerate the disintegration of preparations; therefore, a reasonable combination of both could satisfactorily meet the demand for high-quality 3D printing. γ-Aminobutyric acid can reduce the bitterness of gummy chewable tablets to improve medication compliance and the determined formulation (F7) met the quality requirements. In conclusion, the gum material has excellent potential as an extrusion material for 3D printing. The dosage can be adjusted flexibly by the model shape and size. 3D printing has broad prospects in pediatric preparations.

Full-length transcriptome analysis reveals the differences between floral buds of recessive genic male-sterile line (RMS3185A) and fertile line (RMS3185B) of cabbage.

MAIN CONCLUSION: Starch and sucrose metabolism and plant-pathogen interaction pathways play a dominate role in recessive genic male sterility (RGMS) of cabbage (Brassica oleracea L. var. capitata). RGMS is common in plants and has been widely applied as an effective and economic system for hybrid seed production in many crops. However, little is known regarding the molecular mechanisms of RGMS in cabbage. Hence, full-length transcriptomic and physiological analysis were performed in the spontaneous RGMS mutant RMS3185A and its near-isogenic fertile line (NIL) RMS3185B of small (< 1.6 mm in diameter), medium (~ 2.5 mm in diameter), and large floral buds (~ 3.4 mm in diameter) to identify the differentially expressed genes (DEGs) associated with RMGS. The pollen abnormalities between RMS3185B and RMS3185A appeared at the large floral bud stage. In contrast with RMS3185B, the mature anthers and stamens of RMS3185A were shorter than those of RMS3185B, and the anthers did not dehiscent. The concentrations of glucose, fructose, trehalose, starch, and cellulose in RMS3185A were all significantly lower than those in large floral buds of RMS3185B. PacBio sequencing results showed that DEGs were mainly concentrated in large floral bud stage. In combination with the KEGG enrichment analysis of DEGs in GO terms related to cell wall, pollen and anther, pentose and glucuronate interconversions (ko00040), starch and sucrose metabolism (ko00500), and plant-pathogen interaction (ko04626) were significantly enriched. Among which, cell-wall/pectin-related genes of eighteen PEI, twenty-two PEL, three PG, and fifteen PGL involved in ko00040, and one UGDH, one SPS, four CWINV, four TPP/TPS, and four EGL involved in ko00500, as well as plant-pathogen interaction genes, including sixteen calcium-dependent protein kinase (CDPK), one cyclic nucleotide-gated ion channel (CNGC), and twenty-three calcium-binding protein CML (CML), were significantly down-regulated in RMS3185A relative to that in RMS3185B. Besides, genes involved in ko04626, including two CML and one transcription factor WRKY33, were up-regulated in RMS3185A relative to that in RMS3185B. In conclusion, we hypothesized that the expression alterations of these genes were responsible for calcium signaling and sugar metabolism, thus affecting the occurrence of RGMS in cabbage.

A nanoenzyme linked immunochromatographic sensor for rapid and quantitative detection of SARS-CoV-2 nucleocapsid protein in human blood.

The establishment of a simple, low-cost, high-sensitive and rapid immunoassay for detecting SARS-CoV-2 antigen in human blood is an effective mean of discovering early SARS-CoV-2 infection and controlling the pandemic of COVID-19. Herein, a smartphone based nanozyme linked immunochromatographic sensor (NLICS) for the detection of SARS-CoV-2 nucleocapsid protein (NP) has been developed on demand. The system is integrated by disposable immunochromatography assay (ICA) and optical sensor devices. Immunoreaction and enzyme-catalyzed substrate color reaction were carried out on the chromatographic strip in a device, of which the light signal was read by a photometer through a biosensor channel, and the data was synchronously transmitted via the Bluetooth to the app in-stored smartphone for reporting the result. With a limit of detection (LOD) of 0.026 ng/mL NP, NLICS had the linear detection range (LDR) between 0.05 and 1.6 ng/mL NP, which was more sensitive than conventional ICA. NLICS took 25 min for reporting results. For detection of NP antigen in clinical serum samples from 21 COVID-19 patients and 80 healthy blood donor controls, NLICS and commercial enzyme linked immunosorbent assay (ELISA) had 76.2% or 47.6% positivity, and 100% specificity, respectively (P = 0.057), while a good correlation coefficient (r = 0.99) for quantification of NP between two assays was obtained. In conclusion, the NLICS was a rapid, simple, cheap, sensitive and specific immunochromatographic sensing assay for early diagnosis of SARS-CoV-2 infection.

Antimicrobial ent-abietane-type diterpenoids from the roots of Euphorbia wallichii.

Three previously undescribed ent-abietane-type diterpenoids, designated as 11ß-hydroxy-14-oxo-17-al-ent-abieta-8(9),13(15)-dien-16,12ß-olide (1), 11ß,17-dihydroxy-12-methoxy-ent-abieta-8(14),13(15)-dien-16,12ɑ-olide (2), and 14ɑ-hydroxy-17-al-ent-abieta-7(8),11(12),13(15)-trien-16,12-olide (3), were isolated from 95% ethanol extract of the roots of Euphorbia wallichii. The structures of the new compounds were elucidated by spectroscopic analysis (NMR, IR, UV, and MS). The isolated diterpenoids were tested in vitro for antimicrobial potentials against 6 pathogenic microorganisms. As a result, compounds 1-3 exhibited some antimicrobial activity against the tested Gram positive bacteria with minimum inhibitory concentration values less than 60 µg/ml.[Formula: see text].

[Cloning and tissue expression of mevalonate disphosphate decarboxylase genes from Andrographis paniculata].

Andrographolide is a main bioactive substance in Andrographis paniculata, and extensively used in anti-inflammatory drugs. In order to increase andrographolide production in plant, three 1260 bp ORFs encoding mevalonate disphosphate decarboxylases with 419 amino acids were cloned from A. paniculata by RACE method and analyzed by bioinformatic software. Their tissue expression patterns were predicted by real time PCR. Eleven conserved amino acid residues determining specificity and activity of these MVDs were predicted in these amino acid sequences, but no plastid targeted signal peptides were detected. These MVDs have high similarities with the MVD protein (GenBank number: AEZ55675.1) from Salvia miltiorrhiza. In stems and leaves, expression levels of these MVD genes were constant, and reached the highest level at bud stage and the beginning of flowering. The MVD genes we have cloned from A. paniculata could be used in genetic engineering of andrographolide biosynthsis pathway in future.

Replacing nitrogen in mineral fertilizers with nitrogen in maize straw increases soil water-holding capacity.

Soil water-holding capacity decreases due to long-term mineral fertilizer application. The objective of this study was to determine how replacing mineral fertilizer with maize straw affected the soil water retention curve, soil water content, soil water availability, and soil equivalent pore size. Replacement treatments in which 25% (S25), 50% (S50), 75% (S75), and 100% (S100) of 225 kg ha-1 nitrogen from mineral fertilizer (CK) was replaced with equivalent nitrogen from maize straw were conducted for five years in the Loess Plateau of China. The Gardner model was used to fit the soil water retention curve and calculate the soil water constant and equivalent pore size distribution. The results indicated that the Gardner model fitted well. Replacing nitrogen from mineral fertilizer with nitrogen from straw increased soil specific water capacity, soil readily available water, soil delayed available water, soil available water, soil capillary porosity, and soil available water porosity over time. S25 increased field capacity and wilting point from the fourth fertilization year. S50 enhanced soil readily available water, soil delayed available water, soil available water, and soil available water porosity from the fifth fertilization year, whereas S25 and S75 increased these from the third fertilization year or earlier. Soil specific water capacity, soil readily available water, soil delayed available water, soil available water, soil capillary porosity, and soil available water porosity could better reflect soil water-holding capacity and soil water supply capacity compared with field capacity and wilting point.

Efficacy and safety of add-on antiseizure medications for focal epilepsy: A network meta-analysis.

OBJECTIVE: Several antiseizure medications (ASMs) have been approved for the treatment of focal epilepsy. However, there is a paucity of evidence on direct comparison of ASMs. We evaluated the comparative efficacy and safety of all approved add-on ASMs for the treatment of focal epilepsy using network meta-analysis. METHODS: Data through extensive literature search was retrieved from PubMed, Embase, Cochrane, and ClinicalTrial.gov databases using predefined search terms from inception through March 2023. PRISMA reporting guidelines (CRD42023403450) were followed in this study. Efficacy outcomes assessed were ≥50%, ≥75%, and 100% responder rates. Patient retention rate and safety outcomes such as overall treatment-emergent adverse events (TEAEs) and individual TEAEs were assessed. "Gemtc" 4.0.4 package was used to perform Bayesian analysis. Outcomes are reported as relative risks (RRs) and 95% confidence interval (CI). RESULTS: Literature search retrieved 5807 studies of which, 75 studies were included in the analysis. All ASMs showed significantly higher ≥50% responder rate compared with placebo. Except the ≥75% seizure frequency reduction for zonisamide (2.23; 95% CI: 1.00-5.70) and 100% for rufinamide (2.03; 95% CI: 0.54-11.00), all other interventions showed significantly higher ≥75% and 100% responder rates compared with placebo. Among treatments, significantly higher 100% responder rate was observed with cenobamate compared to eslicarbazepine (10.71; 95% CI: 1.56-323.9) and zonisamide (10.63; 95% CI: 1.37-261.2). All ASMs showed a lower patient retention rate compared to placebo, with the least significant value observed for oxcarbazepine (0.77; 95% CI: 0.7-0.84). Levetiracetam showed a lower risk of incidence (1.0; 95%CI: 0.94-1.1; SUCRA: 0.885067) for overall TEAE compared with other medications. SIGNIFICANCE: All approved ASMs were effective as add-on treatment for focal epilepsy. Of the ASMs included, cenobamate had the greatest likelihood of allowing patients to attain seizure freedom. PLAIN LANGUAGE SUMMARY: This article compares the efficacy and safety of antiseizure medications (ASMs) currently available to neurologists in the treatment of epileptic patients. Several newer generation ASMs that have been developed may be as effective or better than the older medications. We included 75 studies in the analysis. In comparison, all drugs improved ≥50%, ≥75% and 100% responder rates compared to control, except for Zonisamide and Rufinamide in the ≥75% and 100% responder rate categories. Retention of patients undergoing treatment was lower in drugs than placebo. All drugs were tolerated, the levetiracetam showed the best tolerability. Cenobamate more likely help completely to reduce seizures.

A smartphone-based immunochromatographic strip platform for on-site quantitative detection of antigenic targets.

To report the testing signal of an immunochromatographic assay for on-site quantitative detection, a portable and user-friendly smartphone-based biosensing platform is developed in this study. This innovative system is composed of an ambient light sensor inherent smartphone reader and a 3D-printed handhold device, a quantitative tool capable of directly interpreting carbon nanoparticle (CNP)-conjugated immunochromatographic strips. To showcase the platform capability, the smartphone-based immunochromatography system (SPICS) reader and device were successfully used in CNP strips for rapid detection of the early pregnancy marker human chorionic gonadotropin in female urine (HCG; limit of detection [LOD]: 0.30 mIU mL-1), prostate-specific antigen in patient blood (PSA; LOD: 0.28 ng mL-1) and ampicillin residue in animal milk (AMP; LOD: 0.23 ng mL-1). The results were fully correlated with conventional commercial instruments (R2 = 0.99). The SPICS platform exhibits significant advantages, including portability, cost-effectiveness, easy operation, and rapid and quantitative detection, making it a valuable on-site diagnosis tool for use in home and community healthcare facilities.

Carbon nanospheres dual spectral-overlapped fluorescence quenching lateral flow immunoassay for rapid diagnosis of toxoplasmosis in humans.

Toxoplasmosis is a common zoonotic disease caused by a protozoan parasite Toxoplasma gondii (Tox), approximately infecting one-third of human populations worldwide. This study developed the carbon nanospheres (CNPs) based dual spectral-overlapped fluorescence quenching lateral flow immunoassay (CNPs-FQLFIA) for detection of Tox antibodies (ToxAbs). The CNPs have been effectively coupled with Tox antigen (ToxAg), which can completely overlap the excitation and emission spectra of europium nanospheres (EuNPs) and CdSe/ZnS quantum dots (QDs) in testing strips of CNPs-QDs-FQLFIA or CNPs-EuNPs-FQLFIA. The sensitivity of CNPs-EuNPs-FQLFIA or CNPs-QDs-FQLFIA was 4 or 8 IU/mL under natural light readout, or both 4 IU/mL ToxAbs under ultraviolet (UV) light readout by the naked eyes, respectively. The limit of detection (LOD) of two types of CNPs-FQLFIA was both 1 IU/mL ToxAbs under UV light by a dry fluorescence analyzer, but no cross-reaction was found with other antibodies. The intra-assay coefficient variation (CV) of both CNPs-EuNPs-FQLFIA and CNPs-QDs-FQLFIA was less than 8%, while the inter-assay CV was less than 14%, respectively. The correlation coefficient (R2) of CNPs-EuNPs-FQLFIA or CNPs-QDs-FQLFIA to measure the different concentrations of ToxAbs spiked serum samples was 0.99712 and 0.99896, respectively. The CNPs-FQLFIA presented a characteristics of 94.3% sensitivity, 100% specificity and 98% accuracy for detection of ToxAbs in clinical serum samples. In conclusion, CNPs-FQLFIA with EuNPs or QDs fluorescence reporter was an easy, rapid, sensitive, precise and quantitative assay for detecting Tox antibodies in human blood samples.

Ketanserin improves cardiac performance after myocardial infarction in spontaneously hypertensive rats partially through restoration of baroreflex function.

AIM: Baroreflex dysfunction is associated with a higher rate of sudden death after myocardial infarction (MI). Ketanserin enhances baroreflex function in rats. The present work was designed to examine whether ketanserin improves the post-MI cardiac function and to explore the possible mechanism involved. METHODS: Spontaneously hypertensive rats (SHR) were treated with ketanserin (0.3 mg·kg(-1)·d(-1)). Two weeks later, blood pressure and baroreflex function were measured, followed by a ligation of the left coronary artery. The expressions of vesicular acetylcholine transporter (VAChT) and α7 nicotinic acetylcholine receptor (α7-nAChR) in ischemic myocardium, angiogenesis, cardiac function, and left ventricular (LV) remodeling were evaluated subsequently. RESULTS: Ketanserin significantly improved baroreflex sensitivity (0.62±0.21 vs 0.34±0.12 ms/mmHg, P<0.01) and vagal tonic activity (heart rate changes in response to atropine, 54.8±16.2 vs 37.6±13.4 bpm, P<0.01) without affecting the blood pressure or basic heart rate in SHR. Treatment of SHR with ketanserin prominently improved cardiac function and alleviated LV remodeling, as reflected by increases in the ejection fraction, fractional shortening, and LV systolic pressure as well as decreases in LV internal diameter and LV relative weight. The capillary density, vascular endothelial growth factor expression, and blood flow in the ischemic myocardium were significantly higher in the ketanserin-treated group. In addition, ketanserin markedly increased the expression of VAChT and α7-nAChR in ischemic myocardium. CONCLUSION: Ketanserin improved post-MI cardiac function and angiogenesis in ischemic myocardium. The findings provide a mechanistic basis for restoring baroreflex function using ketanserin in the treatment of MI.

Refractory Hypotension in a Late-Onset Mitochondrial Encephalomyopathy, Lactic Acidosis, and Stroke-like Episodes (MELAS) Male with m.3243 A>G Mutation: A Case Report.

(1) Introduction: Symptom spectrum can be of great diversity and heterogeneity in mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) patients in clinical practice. Here, we report a case of MELAS presenting asymptomatic refractory hypotension with m.3243 A>G mutation. (2) Case representation: A 51-year-old male patient presented with a headache, vertigo, and difficulty in expression and understanding. The magnetic resonance imaging of the brain revealed an acute stroke-like lesion involving the left temporoparietal lobe. A definitive diagnosis of MELAS was given after the genetic test identified the chrM-3243 A>G mutation. The patient suffered recurrent stroke-like episodes in the 1-year follow-up. Notably, refractory hypotension was observed during hospitalizations, and no significant improvement in blood pressure was found after continuous use of vasopressor drugs and fluid infusion therapy. (3) Conclusions: We report a case of refractory hypotension which was unresponsive to fluid infusion therapy found in a patient with MELAS. Our case suggests that comprehensive management should be paid attention to during treatment. A further study on the pathological mechanism of the multisystem symptoms in MELAS would be beneficial to the treatment of patients.

First add-on lacosamide treatment in children with focal epilepsy in China: A multi-centre real-life cohort study.

AIM: To assess the effectiveness and safety of lacosamide (LCM) as the first add-on therapy for children with focal epilepsy at multiple centres in China. METHOD: Children aged 4-16 years with focal epilepsy from 13 epilepsy centres were included in this study. All patients were treated with LCM as the first add-on treatment and followed up for 26 weeks. The seizure frequency, seizure-free rate, ≥50% response rate, retention rate, and incidence of adverse drug reactions after the addition of LCM were analysed. RESULTS: Ninety-nine children (58 males; aged 4-16 years; mean age 8.51 ± 2.95 years) were enroled. The mean age at first seizure was 5.74 ± 3.12 years. All patients were administered LCM as the first add-on treatment for focal epilepsy. Twenty-eight patients (28/99, 28.28%) did not experience seizures during the follow-up period. The ≥50% response rates were 80.81% (80/99), 93.88% (92/98), 98.98% (97/98), and 100.0% (98/98) at 6 weeks (visit two), 10 weeks (visit three), 18 weeks (visit four), and 26 weeks (visit five), respectively, compared to that at baseline (visit one). The intelligence scores decreased in 12 participants, remained unchanged in 64, and increased in 16. Adverse events occurred in three participants during the trial, all of which were mild. INTERPRETATION: LCM was effective as the first add-on therapy in this real-life multi-centre study of a paediatric population with focal epilepsy. Further prospective studies with long-term follow-up periods are needed to confirm the effectiveness and tolerability of LCM.

Lacosamide as the first add-on therapy in adult patients with focal epilepsy: A multicenter real-world study.

Background: Prospective observations on the effectiveness, safety, tolerance, and influence of comorbidity of add-on lacosamide (LCM) therapy are still lacking, especially for domestic generic LCM in China. Objective: In this multicenter real-world study, we aimed to evaluate lacosamide (LCM) as the first add-on therapy in adult Chinese patients with focal epilepsy that had initially been treated with monotherapy. Methods: A cohort of consecutive focal epilepsy patients aged over 16 years were enrolled and followed at the multi-epilepsy centers in China. LCM was prescribed as the first add-on therapy. The main outcome measures included seizure frequency and response rate. Data on seizure-free rate, retention rate, scales of depression and anxiety, and adverse events were also collected as additional outcome measures. Results: A total number of 107 adult subjects (60 men, 56.07%) were enrolled. The mean age was 37.16 ± 15.01 years, and the mean age at seizure onset was 312.35 ± 199.97 months. After the LCM add-on therapy, the ≥50% response rates were 76.19, 81.73, 94.12, and 95.79% at the visit at 4 weeks (visit 2), 8 weeks (visit 3), 16 weeks (visit 4), and 24 weeks (visit 5), respectively, compared to the baseline (visit 1). A total of 34 patients (31.78%) had no seizures during the whole follow-up period. The posttreatment emotional performance of the 97 subjects, defined as generalized anxiety disorder (GAD) and Neurological Disorders Depression Inventory (NDDI) scores, was significantly better than the baseline one. Only one patient suffered from mild dizziness. Conclusion: LCM as the first add-on therapy in adult focal epilepsy in China was effective and safe. Further prospective studies with long-term follow-up periods are needed to confirm our present findings. Clinical trial registration: http://www.chictr.org.cn, ChiCTR2100042485.

A Helper Antibody-Based Competitive Fluorescence Immunochromatographic Assay for Quantitative Detection of Florfenicol in Poultry Eggs.

BACKGROUND: Florfenicol (FF) is a chloramphenicol analogue used in animals, and florfenicol amine (FFA) is the main metabolite of FF. However, their residues in agricultural products are harmful to human health. A highly specific and sensitive assay for FF/FFA detection needs to be developed since the traditional detection methods are low in sensitivity. OBJECTIVE: In this study, a new method for rapid quantification of FF/FFA in poultry eggs by helper antibody-based fluorescent immunochromatographic assay (HAFIA) was established. METHODS: Triple antibodies including a primary monoclonal antibody (mAb) specific to the targets FF and FFA, a secondary polyclonal antibody (pAb) labeled with europium nanoparticles (EuNPs), and a helper monoclonal antibody (hAb), reacting with pAb but not with the mAb or the target antigen, are designed, which can form structural aggregation complexes in microwells with a single step of reactions. By loading the reaction sample solution, the triple-antibodies (mAb-pAb-hAb)-EuNPs complexes migrate to the test (T) line on the nitrocellulose membrane of testing strip and are competitively captured by the immobilized FF-bovine serum album (BSA) conjugates on the membrane and the FF/FFA targets in the sample solution. RESULTS: Fluorescence on the T line is read by a portable fluorescent strip reader in 10 min, and the result is given as the ratio of fluorescent intensities on the T and control (C) lines. This new fluorescent testing strip, with amplified signal from the triple-antibody complex, has 50-fold higher sensitivity than conventional colloidal gold-lateral flow immunoassays (CG-LFIAs), and can detect as low as 0.01 ng/mL FF and 0.1 ng/mL FFA targets from egg samples. CONCLUSION: The developed competitive fluorescent immunochromatography method based on auxiliary antibodies has the advantages of high sensitivity and specificity for the rapid and quantitative detection of FF/FFA in poultry eggs. HIGHLIGHTS: Newly designed helper antibody and portable device were applied to quantitative detection. HAFIA tests egg samples and results can be obtained in 10 minutes. HAFIA has the advantages of being more convenient, faster and does not require professional laboratory personnel.

Novel Vertical Flow Immunoassay with Au@PtNPs for Rapid, Ultrasensitive, and On-Site Diagnosis of Human Brucellosis.

Brucellosis is an infectious zoonosis caused by Brucella with clinical symptoms of wavy fever, fatigue, and even invasion of tissues and organs in the whole body, posing a serious threat to public health around the world. Herein, a novel vertical flow immunoassay based on Au@Pt nanoparticles (Au@PtNPs-VFIA) was established for detection of Brucella IgG antibody in clinical serum samples. The testing card of Au@PtNPs-VFIA was manufactured by printing the purified Brucella LPS and goat antimouse IgG on the nitrocellulose membrane as the test-spot or control-spot, respectively. Au@PtNPs labeled with protein G (Au@PtNPs-prG) were concurrently employed as detection probes presenting visible spots and catalysts mimicking catalytic enzymes to catalyze the DAB substrate (H2O2 plus O-phenylenediamine) for deepening color development. The testing procedure of Au@PtNPs-VFIA takes 2-3 min, and the limit of detection (LOD) for Brucella antibody is 0.1 IU/mL, which is faster and more sensitive than that of Au@PtNP-based lateral flow immunoassay (Au@PtNPs-LFIA: 15 min and 1.56 IU/mL, respectively). By comparing with vertical flow immunoassay based on classic Au nanoparticles (AuNPs-VFIA), the Au@PtNPs-VFIA is 32 times or 16 times more sensitive with or without further development of DAB substrate catalysis. Au@PtNPs-VFIA did not react with the serum samples of Gram-negative bacterium infections but only weakly cross-reacted with diagnostic serum of Y. enterocolitica O9 infection. In detection of clinical samples, Au@PtNPs-VFIA was validated for possessing 98.33% sensitivity, 100% specificity, and 99.17% accuracy, which were comparable with or even better than those obtained by the Rose-Bengal plate agglutination test, serological agglutination test, AuNPs-VFIA, and Au@PtNPs-LFIA. Therefore, this newly developed Au@PtNPs-VFIA has potential for rapid, ultrasensitive, and on-site diagnosis of human Brucellosis in clinics.

Characterization of Pre-Existing Neutralizing Antibody to Human Adenovirus Types 5 and 49 and Simian Type 23 in Chinese Population.

Recombinant adenovirus vector has been widely used in vaccine development. Due to the pre-existing immunity of human adenovirus type 5 (HAd5) in humans, a range of rare human and chimpanzee adenovirus vectors have been developed. In the previous study, we constructed novel adenovirus vector Sad23L and Ad49L based on simian adenovirus type 23 (SAd23) and human adenovirus type 49 (HAd49), which were used in the development of ZIKV and COVID-19 vaccines. However, the levels of pre-existing neutralizing antibody (NAb) of HAd49 and SAd23 remain unclear in China. In this study, we measured NAbs titers of HAd5, HAd49, and SAd23 in 600 healthy blood donors from 6 regions across China. NAb titer of HAd49 or SAd23 was significantly lower than that of HAd5 (p < 0.001). There was no significant difference in seroprevalence and NAb titers of three adenoviruses between male and female donors. The seropositive rates of HAd5 and SAd23 increased with age growth in a positive correlation (p < 0.01), while in contrast to HAd5, HAd49, and SAd23 had a low level of pre-existing immunity in Chinese population, which suggested that Ad49L and Sad23L vectors could be used in vaccine development for humans.

Early Phase of Specific Cellular Immune Status Associates with HCV Infection Outcomes in Marmosets.

The major mechanism for determination of HCV infection outcomes has not been fully described, particularly in the early phase of the "window-period" of infection. Based on two groups of marmosets infected with HCV-CE1E2p7/GBV-B chimeric virus (HCV chimera) or GBV-B, the immune mechanism correlating with the different outcomes of virus infections was explored in this study. HCV chimera containing the entire HCV core and envelope proteins (CE1E2p7) and GBV-B RNA were intrahepatically injected into four marmosets in each group, respectively. Blood samples were taken from individual animals in an interval of 2 weeks. Viral load and specific T cell responses were detected in two groups of HCV chimera- and GBV-B-infected marmosets. HCV chimera-infected marmosets appeared to have a virally persistent infection over 6 months post inoculation of the virus. Of these, the specific IFN-γ-secretion T cell response slowly developed over 13 to 19 weeks and was maintained at a relatively low level with 40-70 SFC/106 PBMCs, while the specific Treg cell response was rapidly activated over 3 weeks and was maintained at a high level around 5% among lymphocytes. In contrast, GBV-B-infected marmosets presented spontaneous viral clearance within 6 months; the specific IFN-γ-secretion T cell response was quickly established over 5 to 7 weeks and was maintained at a high level with 50-130 SFC/106 PBMCs, while the specific Treg cell response was inactivated and maintained at a baseline below 3% among lymphocytes. In conclusion, the HCV structural proteins inducing immune suppression in the early phase of HCV infection contributed to the viral persistence, of which the activation of Treg cells might play an important role in the inhibition of an effective T cell antiviral response.

Integrated metabolome and transcriptome analysis reveals salicylic acid and flavonoid pathways' key roles in cabbage's defense responses to Xanthomonas campestris pv. campestris.

Xanthomonas campestris pv. campestris (Xcc) is a vascular bacteria pathogen causing black rot in cabbage. Here, the resistance mechanisms of cabbage against Xcc infection were explored by integrated metabolome and transcriptome analysis. Pathogen perception, hormone metabolisms, sugar metabolisms, and phenylpropanoid metabolisms in cabbage were systemically re-programmed at both transcriptional and metabolic levels after Xcc infection. Notably, the salicylic acid (SA) metabolism pathway was highly enriched in resistant lines following Xcc infection, indicating that the SA metabolism pathway may positively regulate the resistance of Xcc. Moreover, we also validated our hypothesis by showing that the flavonoid pathway metabolites chlorogenic acid and caffeic acid could effectively inhibit the growth of Xcc. These findings provide valuable insights and resource datasets for further exploring Xcc-cabbage interactions and help uncover molecular breeding targets for black rot-resistant varieties in cabbage.