a-toxin-based-counter-selection-system-for-markerless-gene-deletion

A Toxin-Based Counter-Selection System for Markerless Gene Deletion and High-Density Tn5 Transposon Mutagenesis in <em>Pectobacterium brasiliense</em>

Pectobacterium brasiliense is a highly pathogenic bacterium responsible for potato tuber soft rot, blackleg, and aerial stem rot. Elucidation of its ...

We compared changes in morphology and intraocular pressure (IOP) induced by clear 2.2-mm and 3.0-mm corneal incisions in a cohort of patients with cataracts.

Early changes in morphology and intraocular pressure by size of clear corneal incision.

Wir gegen&uuml;ber &Auml;nderungen der Morphologie und Augeninnendruck (IOP), induziert durch Deaktivieren 2,2 mm und 3.0 mm Hornhaut Schnitte in einer Kohorte von Patienten mit grauen Star.

Fr&uuml;he Ver&auml;nderungen der Morphologie und Augeninnendruck durch Gr&ouml;&szlig;e der klare Hornhaut-Schnitt.

Citri Reticulatae Pericarpium (CRP), known as Chenpi (CP) in Chinese, is a medicinal food for health and fitness. In order to find out the characteristic activity chemicals distinguishing various cultivars of CRP and provide a reference for effective development of citrus resources, an "activity fingerprint" of CRP from 21 different cultivars was established based on the evaluation of antitussive and expectorant activities. There were 18 common peaks in the HPLC fingerprint, of which 3 flavonoid glycosides and 14 polymethoxyflavonoids (PMFs) were identified by LC/MS-IT-TOF. Furthermore, five characteristic chemicals were determined and similarity calculation with principal component analysis (PCA) was combined together to compare the similarity and difference among various cultivars. The results showed that some other cultivars were also similar to .  "Chachi" except for .  "Tangerina" and .  "Dahongpao" recorded in Chinese Pharmacopoeia. Most importantly, the peels of .  "Shiyueju," .  "Ponkan," .  "Tribute," and .  "Bayueju," traditionally rarely used for medicinal food, were highly similar to that of .  "Chachi" and rich in bioactive flavonoids, which can be considered the effective medicinal resources of CRP.

Construction and Chemical Profile on "Activity Fingerprint" of Citri Reticulatae Pericarpium from Different Cultivars Based on HPLC-UV, LC/MS-IT-TOF, and Principal Component Analysis.

Mammalian target of rapamycin (mTOR) inhibitor as an attractive drug target with promising antitumor effects has been widely investigated. High quality clinical trial has been conducted in liver transplant (LT) recipients in Western countries. However, the pertinent studies in Eastern world are paucity. Therefore, we designed a clinical trial to test whether sirolimus can improve recurrence-free survival (RFS) in hepatocellular carcinoma (HCC) patients beyond the Milan criteria after LT. This is an open-labeled, single-arm, prospective, multicenter, and real-world study aiming to evaluate the clinical outcomes of early switch to sirolimus-based regimens in HCC patients after LT. Patients with a histologically proven HCC and beyond the Milan criteria will be enrolled. The initial immunosuppressant regimens are center-specific for the first 4-6 weeks. The following regimens integrated sirolimus into the regimens as a combination therapy with reduced calcineurin inhibitors based on the condition of patients and centers. The study is planned for 4 years in total with a 2-year enrollment period and a 2-year follow-up. We predict that sirolimus conversion regimen will provide survival benefits for patients particular in the key indicator RFS as well as better quality of life. If the trial is conducted successfully, we will have a continued monitoring over a longer follow-up time to estimate indicator of overall survival. We hope that the outcome will provide better evidence for clinical decision-making and revising treatment guidelines based on Chinese population data. Trial register: Trial registered at http://www.chictr.org.cn: ChiCTR2100042869.

Efficacy and safety of sirolimus early conversion protocol in liver transplant patients with hepatocellular carcinoma: A single-arm, multicenter, prospective study.

The preparation of biodegradable and antibacterial hydrogels has important clinical value. In this work, a novel strategy has been developed to prepare degradable hydrogel dressings without chemical crosslinking agent using methacrylate anhydride (MA)-modified amylopectin (APMA) and polyacrylamide (PAM). After introducing CC bonds, APMA/PAM hydrogels can be formed under light irradiation. This strategy improves the gelling ability of AP and degradation properties of the hydrogel by avoiding the addition of crosslinking agent. The degradation rate of APMA/PAM hydrogel is 74.04 ± 0.69 % within 12 weeks, while that of APMA/PAM hydrogel containing crosslinking agent is only 38.5 ± 0.1 %. The APMA/PAM hydrogel loading curcumin (Cur) (APMA/PAM-Cur) exhibits high antibacterial efficiency of 98.29 ± 0.41 % and 97.18 ± 0.81 % against S. aureus and E. coli, respectively, with light irradiation. Animal experiments show that the APMA/PAM-Cur hydrogel reduces the infiltration of inflammatory factors, increases the density of collagen, and makes the newly formed granulation tissue thicker and tighter. This study not only proves the promising potential of the APMA/PAM-Cur hydrogel as degradable and antibacterial wound dressing for clinical treatment, but also provides a new strategy for developing low-cost, degradable, and antibacterial wound dressings and reducing antibiotic abuse and environmental pollution caused by medical waste.

Photo-crosslinking methacrylated-amylopectin/polyacrylamide hydrogels loading curcumin for applications as degradable, injectable, and antibacterial wound dressings.

The deficiency of recombinant protein immune response could be compensated for by using nanoparticle platforms or adding immune enhancers, however existing vaccines or adjuvants struggle to elicit durable cellular immune responses. In this work, a protein nanoscaffold, lumazine synthase isolated from Brucella (BLS) was optimally designed that could facilitate cellular uptake of displayed antigens and the maturation of bone marrow-derived dendritic cells (BMDCs), and enhancing humoral immune responses. To enhance cellular immune response, chitosan hydrochloride-stabilized Pickering emulsion (CHSPE) was evaluated as an adjuvant for the BLS nanoscaffold-based vaccine. CHSPE could enhance the recruitment and activation of DCs at the injection site and the uptake of antigen and activation of DCs in the draining lymph nodes (dLNs), compared with commercial adjuvant ISA 206. In addition, CHSPE induced antigen-specific antibody levels comparable to those of ISA 206 and increased the ratio of central memory T cells (TCM) and effector memory T cells (TEM), and promoted the secretion of Th1-type cytokines. Moreover, CHSPE-formulated vaccine provided a similar level of protection to the live attenuated vaccine and was superior to that of ISA 206. Taken together, the combination of the BLS nanoscaffold and CHSPE provides a feasible strategy for recombinant protein subunit vaccine development.

Combination application of Pickering emulsion and BLS protein nanoparticles enhances vaccine efficacy.

Brucellosis presents notable diagnostic challenges, particularly during the chronic phase of infection, due to the typically low bacterial load in tissues, which may evade detection by conventional Polymerase Chain Reaction (PCR) methods. Traditional PCR techniques are susceptible to cross-reactions and can produce false-positive or false-negative results. Furthermore, bacteriological and serological tests have recognized limitations in sensitivity and specificity, potentially complicating accurate diagnosis. We developed a novel reverse transcription quantitative PCR (RT-qPCR) assay targeting Brucella species' highly abundant and conserved 23S rRNA gene to address these challenges. The 23S rRNA gene was selected as an optimal molecular target due to the approaches' number and genetic stability across Brucella strains. Primers were designed based on a conserved region (Sequence ID: NR_103168.2) to ensure broad applicability, and their specificity was verified through BLAST analysis. A key feature of the 23S-RT-qPCR method is incorporating a reverse transcription step to convert RNA into complementary DNA (cDNA), which may enhance detection sensitivity. Using the Brucella S2 vaccine strain as a template, experimental results indicated that the 23S-RT-qPCR method reduced cycle threshold (Ct) values by approximately 2-3 units (14.67-16.74) compared to conventional qPCR. Statistical analysis using a T-test showed that this reduction was significant (P < 0.05). The performance of the 23S-RT-qPCR method was further evaluated using clinical samples and compared with the IS711 detection method. The 23S-RT-qPCR method demonstrated a lower false-positive rate (2.6%) and false-negative rate (7.6%) compared to the IS711 method, which had a false-positive rate of 5.2% and a false-negative rate of 7.6%. These results suggest that the 23S-RT-qPCR method offers improved sensitivity and specificity, reducing false-positive and false-negative rates in clinical sample analysis. Overall, this method may provide a more reliable approach for diagnosing chronic brucellosis and could have broader applicability in detecting other bacterial pathogens.

Establishment of a 23S rRNA assay for Brucella and its application in evaluating bacterial growth status.

Systemic lupus erythematosus (SLE) is characterized by the production of pathogenic autoantibodies, particularly antidouble-stranded (ds) DNA antibodies, which contribute to multiorgan damage (lupus nephritis, LN). Hence, there is an urgent need to recognize and eliminate SLE-specific anti-ds DNA antibodies to enhance the SLE treatment. Herein, mesoporous silica (MSNs) loaded with SeC and surface-modified ctDNA are constructed to effectively specific bind and eliminate pathogenic anti-dsDNA antibodies for treatment SLE in 125 plasm and enabling swift LN diagnosis in 36 kidney tissue from SLE patients. As expected, the clearance ratio of anti-dsDNA antibodies by nanotherapeutics is significantly greater compared to other products commonly used in clinical therapies and exhibits biocompatibility and safety in patients. Moreover, MSNs-DNA can also help visualize the distribution of anti-dsDNA antibodies in the lesions of the kidney. Importantly, the combination strategy (MSNs-DNA@SeC) can effectively remove antibodies and reduce UP production by the regulation of B cells and T cells in female MRL/lpr SLE model mice to alleviate related symptoms. Collectively, the resultant data not only presents a straightforward method for the systematic design of nanomedicine targeting SLE to enhance the effects on diagnosis and treatment, but also elucidates the potential mechanisms involving anti-dsDNA antibodies in the pathogenesis and progression of SLE.

Targeting Nanotherapeutics for Highly Efficient Diagnosis and Treatment of Systemic Lupus Erythematosus through Regulation of Immune Response.

Brucellosis is a zoonotic infectious disease caused by Brucella species, posing serious threats to public health and livestock industries worldwide. According to the World Health Organization (WHO), over one million new cases are reported annually. However, the actual number may be underestimated due to the bacteria's intracellular persistence and subclinical bacteremia. These features and Brucella's genetic variability and immune evasion strategies hinder accurate diagnosis and effective treatment. Thus, there is an urgent need for sensitive and specific molecular probes. This study used whole Brucella bacteria as the target antigen for aptamer selection via the Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method, combined with a microplate-based screening approach. To enhance specificity, a multi-organism counter-selection strategy was implemented using Rhizobium fischeri, Escherichia coli, and Salmonella. After 13 rounds of selection, a high-affinity aptamer library was obtained. Candidate aptamers were cloned, sequenced using T-vector cloning, and analyzed phylogenetically. Five candidates (WS-6, WS-17, WS-44, WS-26, and WS-32) were selected for further characterization. Binding affinity and specificity assays identified WS-6 as the top-performing aptamer (Kd = 16.23 ± 5.84 nM). WS-6 also demonstrated thermal and pH stability. Its target protein was identified as the outer membrane protein BamA (β-barrel assembly machinery A) using Co-immunoprecipitation (Co-IP), mass spectrometry, and molecular docking. Western blotting and pull-down assays confirmed this interaction. Functional studies showed that WS-6-loaded liposomes selectively recognized Brucella and significantly reduced bacterial invasion in host cells. qPCR analysis demonstrated a notable decrease in bacterial load in WS-6-treated cells (Ct values from 22.34 to 20.64, P < 0.05). Mutational analysis of WS-6 confirmed the importance of GC-rich binding sites. Moreover, a truncated version of BamA (BamAK) retained binding capability, validating it as the key antigenic domain. In summary, this study demonstrates the potential of SELEX-derived aptamers as diagnostic and therapeutic tools for Brucella infection. Identifying BamA as a target provides a molecular basis for the future development of precise detection methods and targeted therapies against brucellosis.

Screening and characterization of nucleic acid aptamers targeting Brucella surface antigens: identification and analysis of their corresponding binding sites.

In heart failure (HF), acute kidney injury (AKI) is an independent risk factor of mortality, and severer stage of AKI is associated with poorer prognosis. Blood pressure response index (BPRI) reflects the cardiovascular response to vasoactive drugs which are commonly used to increase perfusion to the vital organs. We sought to investigate the association between BPRI and AKI progression in HF patients.

Blood pressure response index and acute kidney injury progression in heart failure patients: a retrospective cohort study from MIMIC-IV.

Goniotomy (GT) or ab interno trabeculotomy facilitates aqueous humor drainage thereby resulting intraocular pressure (IOP) reduction by incising the trabecular meshwork and the inner wall of Schlemm's canal. It can be performed alone or combined with cataract surgery, a procedure known as phacogoniotomy. As these procedures can be performed with non-device built instruments compared to other minimally invasive glaucoma surgery (MIGS) devices or procedures, they are increasingly adopted in low- and middle-income countries (LMICs) and primary-level hospitals. To summarize the clinical use and future directions of GT and phacogoniotomy, we conducted a comprehensive literature review by searching PubMed. The available evidence suggests that GT and phacogoniotomy are effective and safe procedures, providing a widely adopted surgical option applicable to glaucoma patients of all ages.

Goniotomy and phacogoniotomy in glaucoma surgery.

Aortic stenosis (AS) is a prevalent heart valve disease; however, morbidity and mortality are significantly reduced by aortic valve replacement (AVR). The European System for Cardiac Operative Risk Evaluation II (EuroSCORE II) is used to assess perioperative mortality risk in patients with severe AS undergoing AVR. This study aimed to evaluate the prognostic value of EuroSCORE II for long-term all-cause mortality in Chinese patients with moderate-to-severe AS, determine whether AVR affects this prognostic value, and identify the best cut-off value for low-risk EuroSCORE II patients without AVR.

Evaluation of EuroSCORE II to Determine the Prognosis of Patients With Moderate-To-Severe Aortic Stenosis: A Long-Term Retrospective Study.

To evaluate the efficacy of intraocular pressure (IOP)-lowering medications in slowing down axial elongation among adults with progressive high myopia (PHM).

Intraocular Pressure-Lowering Therapy Slows Down Axial Elongation in Progressive High Myopia with Enhanced Choroidal Perfusion.

To evaluate the visual preservation, safety, and surgical outcomes of phacogoniotomy (phacoemulsification combined with or without goniosynechialysis and goniotomy) in patients with end-stage glaucoma.

Article	Year
Early changes in morphology and intraocular pressure by size of clear corneal incision. Cornea\| PubMed ID: 21282997	2011
Construction and Chemical Profile on "Activity Fingerprint" of Citri Reticulatae Pericarpium from Different Cultivars Based on HPLC-UV, LC/MS-IT-TOF, and Principal Component Analysis. Evidence-based complementary and alternative medicine : eCAM\| PubMed ID: 32190084	2020
Efficacy and safety of sirolimus early conversion protocol in liver transplant patients with hepatocellular carcinoma: A single-arm, multicenter, prospective study. Hepatobiliary & pancreatic diseases international : HBPD INT\| PubMed ID: 34583911	2022
Photo-crosslinking methacrylated-amylopectin/polyacrylamide hydrogels loading curcumin for applications as degradable, injectable, and antibacterial wound dressings. International journal of biological macromolecules\| PubMed ID: 39154693	2024
Combination application of Pickering emulsion and BLS protein nanoparticles enhances vaccine efficacy. International journal of biological macromolecules\| PubMed ID: 39547601	2024
Establishment of a 23S rRNA assay for Brucella and its application in evaluating bacterial growth status. Veterinary research communications\| PubMed ID: 39982617	2025
Targeting Nanotherapeutics for Highly Efficient Diagnosis and Treatment of Systemic Lupus Erythematosus through Regulation of Immune Response. Small science\| PubMed ID: 40395345	2025
Screening and characterization of nucleic acid aptamers targeting Brucella surface antigens: identification and analysis of their corresponding binding sites. World journal of microbiology & biotechnology\| PubMed ID: 40451960	2025
Blood pressure response index and acute kidney injury progression in heart failure patients: a retrospective cohort study from MIMIC-IV. BMC nephrology\| PubMed ID: 41390635	2025
Goniotomy and phacogoniotomy in glaucoma surgery. Asia-Pacific journal of ophthalmology (Philadelphia, Pa.)\| PubMed ID: 41786220	2026
Evaluation of EuroSCORE II to Determine the Prognosis of Patients With Moderate-To-Severe Aortic Stenosis: A Long-Term Retrospective Study. Reviews in cardiovascular medicine\| PubMed ID: 41789337	2026
Intraocular Pressure-Lowering Therapy Slows Down Axial Elongation in Progressive High Myopia with Enhanced Choroidal Perfusion. Ophthalmology science\| PubMed ID: 42027466	2026
Visual preservation and surgical outcomes of phacogoniotomy in end-stage glaucoma: A multicenter study. Asia-Pacific journal of ophthalmology (Philadelphia, Pa.)\| PubMed ID: 42167481	2026

Bin Xie

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