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Biomedical subjects

Lijuan Li

Publications and source records attributed to Lijuan Li.

9 recordsLinked to original sources

Engineered genomic attachment sites for site-specific recombinases enable high-efficiency integration in plants and human cells.

Here we developed a DNA-centric strategy for optimizing site-specific recombination by rationally engineering chimeric attachment sites. The high-activity att variants enhance Bxb1-mediated integration efficiency in human cells and plants. Among these att variants, the engineered attB(V111) site achieved 51.9% integration efficiency in HEK293T cells (1.7-fold versus wild-type attB) and 35.6% in rice protoplasts (4.4-fold versus wild-type attB). When paired with an engineered single protein mutant in the Bxb1 catalytic domain, the optimized system achieved targeted integration efficiencies of 31% for a CD19 chimeric antigen receptor cassette and 25% for an ornithine transcarbamylase expression cassette in human cells. In rice, these engineered variants enabled integration of a 5.8 kb herbicide-resistance cassette at a targeted genomic locus, with stable integration detected in 24% of regenerated plants. Oxford Nanopore-based long-read sequencing of edited plants reveals complete and precise insertion with high specificity. Propagation of edited seedlings to T1 plants confirms heritable editing to future generations. This approach provides a safe, broadly applicable approach for recombinase-based genome editing.

Journal Article↗

CaCl2 Priming Boosts Salinity-Alkalinity Tolerance in Germinating Soybean by Reducing DNA Oxidative Damage and Enhancing Ca2+ -ROS Signaling Crosstalk.

Soybean (Glycine max) seed germination is highly sensitive to saline-alkaline stress. Seed priming represents an effective strategy to mitigate its detrimental effects. However, the optimal priming conditions (agent, concentration, duration) and the underlying molecular mechanisms remain poorly understood. This study investigated the effects of priming with distilled water (Control), calcium chloride (CaCl2), melatonin (MT), and proline (Pro) under saline-alkaline stress on soybean seed germination and the molecular basis of enhanced tolerance. Evaluation of ten germination-related parameters revealed that priming with 100 mM CaCl2 for 12 h significantly enhanced the germination rate. Physiological analyses demonstrated that CaCl2 priming effectively reduced reactive oxygen species (ROS) accumulation by increasing the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), while decreasing malondialdehyde (MDA) content. Furthermore, CaCl2 priming activated the Ca2+ signaling pathway by increasing radicle Ca2+ content and upregulating the expression levels of Ca2+ signaling-related genes (e.g., GmCAM7, GmCNGC2, GmCNGC19, GmMPK2, and GmMKK2). Additionally, CaCl2 priming significantly enhanced DNA damage repair capacity of soybean cultivars with differing saline-alkaline tolerance. This was manifested by reduced DNA oxidative damage and decreased random amplified polymorphic DNA (RAPD) polymorphism, thereby enhancing genomic stability and alleviating cell cycle arrest. These findings deepen our understanding of the complex regulatory role of calcium signaling in plant abiotic stress responses and provide important novel theoretical insights for improving crop resilience.

Glycine max↗

Quantitative relationship of two viruses (MrNV and XSV) in white-tail disease of Macrobrachium rosenbergii.

Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) were purified from diseased freshwater prawns M. rosenbergii and used to infect healthy post-larvae (PL) by an immersion method. Three groups of prawns were challenged with various combined doses of MrNV and XSV. Signs of white-tail disease (WTD) were observed in Groups 1 and 2, which had been challenged with combinations containing relatively high proportions of MrNV and low proportions of XSV. By contrast there was little sign of WTD in Group 3, which had been challenged with a higher proportion of XSV than MrNV. A 2-step Taqman real-time RT-PCR was developed and applied to quantify viral copy numbers in each challenged PL. Results showed that genomic copies of both viruses were much higher in Groups 1 and 2 than they were in Group 3, indicating that MrNV plays a key role in WTD of M. rosenbergii. The linear correlation between MrNV and XSV genome copies in infected prawns demonstrated that XSV is a satellite virus, dependent on MrNV, but its role in pathogenicity of WTD remains unclear.

Animals↗

Design, synthesis, and photochemical behavior of poly(benzyl ester) dendrimers with azobenzene groups throughout their architecture.

A new class of dendrons and dendrimers containing azobenzene units (bearing up to 29 azobenzene groups for four generations) were designed and synthesized with the convergent method, which uses azobenzene derivatives as monomers and benzyl ester groups as linkages leading to photoresponsive dendrons and dendrimers with azobenzene units throughout their architecture. Photochemical isomerization experiments revealed that all of the dendrons and dendrimers undergo trans-cis isomerization by irradiation and cis-trans isomerization by either irradiation or heating.

Azo Compounds↗

Partial and delayed release of troponin-I compared with the release of lactate dehydrogenase from necrotic cardiomyocytes.

Although the troponins are the serum proteins most frequently used nowadays to diagnose myocardial infarction, controversy continues about whether troponins are released later from infarcted myocardium than the cytoplasmic enzymes used previously, like lactate dehydrogenase (LDH). The present study compared the release kinetics of troponin-I (TnI) and LDH from necrotic cardiomyocytes in vitro. Cardiomyocytes prepared from neonatal rat ventricles were grown for 3 days. A total of 126 cultures were subjected to metabolic inhibition to induce cell necrosis. At various time intervals cells and media were collected for quantitative analysis of LDH activity and TnI concentration. Mean (+/-SD) LDH activity and TnI content of nine cultures at time t=0 were 2.07+/-0.30 U and 1.52+/-0.30 micro g per culture, respectively. Release of LDH from necrotic cardiomyocytes preceded release of TnI by about 60 min. The quantity of LDH released from the cultures after 210 min was 83.2+/-10.0%, whereas that of TnI released after 210 min was always less (33.8+/-22.2%). Cytochemical assessment of necrotic cardiomyocytes showed TnI-positive cells that were poor in LDH. The delay of TnI release relative to LDH release may be explained by slow dissociation of TnI molecules from myofilaments and/or formation of TnI degradation products that are undetected by the currently used ELISA assay.

Animals↗

Synthesis, structure, and catalytic activity of mononuclear iron and (mu-Oxo)diiron complexes with the ligand 2,6-bis(N-methylbenzimidazol-2-yl)pyridine.

Iron complexes including polyimidazole and exchangeable ligands are studied with the aim of modeling the structural and functional features of the non-heme iron centers of dinuclear proteins, such as methane monooxygenase. In [Fe(2)OL(2)(MeOH)(2)(NO(3))(2)](NO(3))(2) (1) (L = 2,6-bis(N-methylbenzimidazol-2-yl)pyridine), each Fe(III) is in a distorted octahedral environment and has a donor set of N(3)O(3) which includes three N atoms from L and three O atoms from a nitrate, micro-oxo, and methanol. In complex [FeLCl(3)] (2) (L = 2,6-bis(N-methylbenzimidazol-2-yl)pyridine), Fe(III) is coordinated to three nitrogen atoms from L and three chloride ions. Complex 1 efficiently catalyzed the oxidation of cyclohexane with 51% conversion to cyclohexanol. It also catalyzed the epoxidation of styrene, cyclohexane, 2-methyl-2-butene, and cis- and trans-2-heptene with 51-84% conversions and high selectivity (71-99%) for epoxide products. Complex 2, however, has no specific reactivity toward these substrates. From the alcohol/ketone (A/K) ratio close to 1 in the oxidation of cyclohexane, the low KIE (kinetic isotope effect K(H)/K(D) ratio = 1.8) for cyclohexanol formation, and the nonstereospecificity of the oxidation of cis-dimethylcyclohexane, it can be concluded that long-lived alkyl radicals are involved in the oxidation catalyzed by complex 1. On the other hand, the stereospecific epoxidation of alkenes, the stereoselective oxidation of cumene, and the high degree of retention of configuration in the oxidation of cis- and trans-2-heptene suggest that a nonradical species, probably a metal-based intermediate, is involved in the oxidation of alkenes and cumene.

Benzimidazoles↗

Microfluidic device for capillary electrochromatography-mass spectrometry.

A novel microfabricated device that integrates a monolithic polymeric separation channel, an injector, and an interface for electrospray ionization-mass spectrometry detection (ESI-MS) was devised. Microfluidic propulsion was accomplished using electrically driven fluid flows. The methacrylate-based monolithic separation medium was prepared by photopolymerization and had a positively derivatized surface to ensure electroosmotic flow (EOF) generation for separation of analytes in a capillary electrochromatography (CEC) format. The injector operation was optimized to perform under conditions of nonuniform EOF within the microfluidic channels. The ESI interface allowed hours of stable operation at the flow rates generated by the monolithic column. The dimensions of one processing line were sufficiently small to enable the integration of 4-8 channel multiplexed structures on a single substrate. Standard protein digests were utilized to evaluate the performance of this microfluidic chip. Low- or sub-fmol amounts were injected and detected with this arrangement.

Electrophoresis, Capillary↗

LPS-induced NF-kappa B activation requires Ca2+ as a mediator in isolated pancreatic acinar cells of rat.

OBJECTIVE: To investigate the effect of Ca(2+) on lipopolysaccharide (LPS)-induced NF-kappa B activation in pancreatic acinar cells and the role of NF-kappa B in LPS-induced acinar cell injury. METHODS: Male rat pancreatic acinar cells were isolated by collagenase digestion, then exposed to varying concentrations of LPS (from 1 to 20 mg/L) in the presence or absence of EGTA. At various time points (30 minutes, 1 hour, 2 hours, 4 hours and 10 hours) after treatment with the agents, cell viability was determined by MTT. Nuclear translocation of NF-kappa B's subunit p65 was visualized by immunofluorescence staining and nuclei protein was extracted to perform EMSA which was used to assay the activity of NF-kappa B binding to the DNA sequence containing the recognition site of NF-kappa B. RESULTS: LPS induced cell damage in a time- and concentration-dependent manner while EGTA attenuated LPS-induced cell damage (P < 0.05). NF-kappa B p65 immunofluorescence staining had increased intensity in the cytoplasm and indicated that nuclear translocation occurred within 30 minutes and its zenith was reached at 1 hour after LPS (10 mg/L) treatment. Testing of NF-kappa B DNA binding activity showed the same alteration phase as p65 immunofluorescence staining. NF-kappa B activation preceded the pathological alteration of pancreatic acinar cells. The Ca(2+) chelator EGTA inhibited LPS-induced NF-kappa B activation. CONCLUSIONS: NF-kappa B activation is an important early event in LPS-induced injury to pancreatic acinar cells. Ca(2+) is an important mediator in the process of LPS-induced NF-kappa B activation.

Animals↗