Prenatal diagnosis of interrupted aortic arch using high-definition flow render mode and spatiotemporal image correlation.

OBJECTIVES: To evaluate the clinical utility of two dimensional (2D) ultrasound combined with spatiotemporal image correlation (STIC) in diagnosing interrupted aortic arch (IAA) in fetal life. METHODS: A total of 53 cases of fetal IAA were diagnosed using 2D ultrasound combined with STIC, and 53 normal fetuses of the same gestational week were selected. These cases were retrospectively analyzed to assess the utility of employing 2D ultrasound combined with STIC in the diagnosis of IAA. RESULTS: 2D ultrasound combined with STIC detected 22 cases of type A IAA, 24 cases of type B IAA, and seven cases of type C IAA. Furthermore, combining 2D ultrasound with STIC enabled dynamic visualization of the IAA, aiding in prenatal diagnosis. The diagnostic coincidence rate of IAA was found to be higher in the HD-flow combined with STIC than that in the 2D combined with HD-flow. CONCLUSION: HD-flow combined with STIC can assist in diagnosing fetal IAA, and this technique has important clinical value.

Protective effects of ferulic acid on embryonic development by improving antioxidant function in broilers embryo of thermal manipulation.

This study aimed to elucidate the effects of broiler embryos soaked in ferulic acid (FA) solution on alleviating the negative impact of thermal manipulation (TM) on chicken embryo development and to provide a theoretical and experimental basis for applying TM and FA in the poultry feeding industry. A total of 120 broiler fertilized eggs were randomly divided into three groups: control group, TM group, and comprehensive group (TM + FA), with 40 eggs in each group. The TM group and the comprehensive group from the 7th embryonic age to the 16th embryonic age received TM for ten days, treated with a temperature of 39.5 °C and relative humidity of 65% for 18 h a day. In the comprehensive group, broiler embryos were immersed in FA solution at a concentration of 80 mg/L for 6 min at 16:00 every day from the 6th to the 8th embryo age. They were incubated continuously after being soaked until the chicks hatched. The results showed that the rates of dead embryos and weak chicks in the TM group were significantly higher than those in the control group and comprehensive group. Chick body temperatures of the TM group and comprehensive group were significantly lower than those of the control group. The heart weights of the TM group and comprehensive group were significantly lower than those of the control group, and the leg weights of the TM group were significantly decreased compared with those of the control group and comprehensive group. The SOD activity of serum in the comprehensive group was significantly higher than that in the control group and TM group, while the CAT activity of serum in the comprehensive group and control group was significantly higher than that in the TM group; however, there was no difference between the comprehensive group and control group. The activities of SOD and CAT in the liver were significantly higher than those of the TM group; however, the MDA content of the liver in the comprehensive group and control group was significantly lower than that of the TM group. The gene expression of Nrf2 and SOD in the comprehensive group and TM group was significantly higher than that in the control group; however, there was no significant difference between the comprehensive group and TM group. Soaking broiler embryonic eggs in an FA solution can improve the antioxidant capacity of the liver by upregulating Nrf2-Keap1 signal pathway-related gene expression. FA can effectively alleviate the side effects of TM on chicken embryos and does not impact the effects of TM.

Redox Electrolytes-Assisting Aqueous Zn-Based Batteries by Pseudocapacitive Multiple Perovskite Fluorides Cathode and Charge Storage Mechanisms.

Aqueous Zn-based batteries (AZBs) have attracted intensive attention. However, to explore advanced cathode materials with in-depth elucidation of their charge storage mechanisms, improve energy storage capacity, and construct novel cell systems remain a great challenge. Herein, a new pseudocapacitive multiple perovskite fluorides (ABF3 ) cathode is designed, represented by KMF-(IV, V, and VI; M = NiCoMnZn/-Mg/-MgFe), and constructed Zn//KMF-(IV, V, and VI) AZBs and their flexible devices. Ex situ tests have revealed a typical bulk phase conversion mechanism of KMF-VI electrode for charge storage in alkaline media dominated by redox-active Ni/Co/Mn species, with transformation of ABF3 nanocrystals into amorphous metal oxide/(oxy)hydroxide nanosheets. By employing single or bipolar redox electrolyte strategies of 20 mm [Fe(CN)6 ]3- or/and 10 mm SO3 2- /Cu[(NH3 )4 ]2+ acting on KMF-(IV, V, and VI) cathode and Zn anode, the AZBs show an improved energy storage owing to additional capacity contribution of redox electrolytes. The as-designed Zn//polyvinyl alcohol (PVA)-KOH-K3 [Fe(CN)6 ]//KMF-(IV, V, and VI) redox gel electrolytes-assisting flexible AZBs (RGE-FAZBs) exhibit remarkable performance under different bending angles because of slight dissolution corrosion of zinc anode compared with liquid electrolytes. Overall, the work demonstrates the novel idea of conversion-type multiple ABF3 cathode for redox electrolytes-assisting AZBs (RE-AZBs) and their flexible systems, showing great significance on electrochemical energy storage.

Direct capture and amplification of nucleic acids using a universal, elution-free magnetic bead-based method for rapid pathogen detection in multiple types of biological samples.

Nucleic acid amplification tests (NAATs) have become an attractive approach for pathogen detection, and obtaining high-quality nucleic acid extracts from biological samples plays a critical role in ensuring accurate NAATs. In this work, we established an elution-free magnetic bead (MB)-based method by introducing polyethylene-polypropylene glycol (PEPPG) F68 in lysis buffer and using NaOH solution instead of alcohols as the washing buffer for rapid nucleic acid extraction from multiple types of biological samples, including nasopharyngeal swabs, serum, milk, and pork, which bypassed the nucleic acid elution step and allowed the nucleic acid/MB composite to be directly used as the template for amplification reactions. The entire extraction process was able to be completed in approximately 7 min. Even though the nucleic acid/MB composite could not be used for quantitative real-time PCR (qPCR) assays, this elution-free MB-based method significantly improved the sensitivity of the loop-mediated isothermal amplification (LAMP) assay. The sensitivity of the quantitative real-time LAMP (qLAMP) assays combined with this elution-free MB-based method showed an improvement of one to three orders of magnitude compared with qLAMP or qPCR assays combined with the traditional MB-based method. In addition to manual operation, like the traditional MB-based method, this universal, rapid, and facile nucleic acid extraction method also has potential for integration into automated robotic processing, making it particularly suitable for the establishment of an analysis platform for ultrafast and sensitive pathogen detection in various biological samples both in centralized laboratories and at remote sites.

Comparative study on the gene expression of corticosterone metabolic enzymes in embryonic tissues between Tibetan and broiler chickens.

Glucocorticoids (GCs) are an essential mediator hormone that can regulate animal growth, behavior, the phenotype of offspring, and so on, while GCs in poultry are predominantly corticosterones. The biological activity of GCs is mainly regulated by the intracellular metabolic enzymes, including 11ß-hydroxysteroid dehydrogenases 1 (11ß-HSD1), 11ß-hydroxysteroid dehydrogenases 2 (11ß-HSD2), and 20-hydroxysteroid dehydrogenase (20-HSD). To investigate the embryonic mechanisms of phenotypic differences between breeds, we compared the expression of corticosterone metabolic enzyme genes in the yolk-sac membrane and chorioallantoic membrane (CAM). We described the tissue distribution and ontogenic patterns of corticosterone metabolic enzymes during embryonic incubation between Tibetan and broiler chickens. Forty fertilized eggs from Tibetan and broiler chickens were incubated under hypoxic and normoxic conditions, respectively. Real-time fluorescence quantitative PCR was used to examine the expression of 11ß-HSD1/2, and 20-HSD mRNA in embryonic tissues. The results showed that the expression levels of yolk-sac membrane mRNA of 11ß-HSD2 and 20-HSD in Tibetan chickens on E14 (embryonic day of 14) were significantly lower than those of broiler chickens (P < 0.05), and these genes expression of CAM in Tibetan chickens were higher than those of broiler chickens (P < 0.05). In addition, the three genes in the yolk-sac membrane and CAM were followed by a down-regulation on E18 (embryonic day of 18). The 11ß-HSD1 and 11ß-HSD2 genes followed a similar tissue-specific pattern: the expression level was more abundantly in the liver, kidney, and intestine, with relatively lower abundance in the hypothalamus and muscle, and the expression level of 20-HSD genes in all tissues tested was higher. In the liver, 20-HSD of both Tibetan and broiler chickens showed different ontogeny development patterns, and hepatic mRNA expression of 20-HSD in broiler chickens was significantly higher than that of Tibetan chickens of the same age from E14 to E18 (P < 0.05). This study preliminarily revealed the expression levels of cortisol metabolic genes in different tissues during the development process of Tibetan and broiler chicken embryos. It provided essential information for in-depth research of the internal mechanism of maternal GCs programming on offspring.

Clinical value of prenatal ultrasound in diagnosis and classification of common arterial trunk.

OBJECTIVES: To investigate the clinical value of prenatal ultrasound in the diagnosis of the common arterial trunk (CAT) classification and associated malformations. MATERIALS AND METHODS: The 2D ultrasound images, spatiotemporal image correlations (STICs) and clinical data of 88 fetuses diagnosed with CAT malformations by prenatal ultrasound were retrospectively analyzed and classified. The correlation between different types, fetal malformation and pregnancy outcomes were analyzed. RESULTS: Among the 88 fetuses, there were 39 cases (44.32%) of type A1, 40 cases (45.45%) of type A2, 8 cases (9.09%) of type A3, and 1 case of type A4 (1.14%). There were 16 cases (18.18%) with isolated CAT, 48 cases (54.55%) with complex intra-cardiac structural abnormalities, and 24 cases (27.27%) with intra-cardiac and extra-cardiac structural abnormalities. In extra-cardiac structural malformations, 14 cases were associated with 1 other system abnormality, 4 cases with 2 other system abnormalities, 3 cases with 3 other system abnormalities, while 3 cases were combined with 4 other system abnormalities, among which the facial and physical abnormalities had the highest incidence (39.13%). The STIC images were completely displayed in all 88 cases. There was a statistical difference between isolated CAT and CAT combined with other abnormalities in fetal pregnancy outcomes. CONCLUSIONS: Prenatal ultrasound had a high clinical application value in CAT classification. Pregnancy outcomes were highly correlated with the classification and associated intra-cardiac and extra-cardiac structural malformations. The early evaluation of fetal prognosis before birth has important value for clinical intervention.

[Esophageal Angiolipoma:Report of One Case and Literature Review].

Esophageal angiolipoma is a rare disease with unspecific clinical manifestations.This paper reported a case of esophageal angiolipoma confirmed by upper gastrointestinal endoscopy and summarized the clinical manifestations,endoscopic and pathological features,treatment and prognosis of the patients by reviewing the relevant literature,aiming to provide references for clinical diagnosis and treatment of this disease in the future.

A single N-terminal phosphomimic disrupts TDP-43 polymerization, phase separation, and RNA splicing.

TDP-43 is an RNA-binding protein active in splicing that concentrates into membraneless ribonucleoprotein granules and forms aggregates in amyotrophic lateral sclerosis (ALS) and Alzheimer's disease. Although best known for its predominantly disordered C-terminal domain which mediates ALS inclusions, TDP-43 has a globular N-terminal domain (NTD). Here, we show that TDP-43 NTD assembles into head-to-tail linear chains and that phosphomimetic substitution at S48 disrupts TDP-43 polymeric assembly, discourages liquid-liquid phase separation (LLPS) in vitro, fluidizes liquid-liquid phase separated nuclear TDP-43 reporter constructs in cells, and disrupts RNA splicing activity. Finally, we present the solution NMR structure of a head-to-tail NTD dimer comprised of two engineered variants that allow saturation of the native polymerization interface while disrupting higher-order polymerization. These data provide structural detail for the established mechanistic role of the well-folded TDP-43 NTD in splicing and link this function to LLPS. In addition, the fusion-tag solubilized, recombinant form of TDP-43 full-length protein developed here will enable future phase separation and in vitro biochemical assays on TDP-43 function and interactions that have been hampered in the past by TDP-43 aggregation.

Influence of fish skin gelatin-sodium alginate complex stabilized emulsion on benzyl isothiocyanate stability and digestibility in vitro and in vivo.

BACKGROUND: An emulsion delivery system for benzyl isothiocyanate (BITC) was prepared using fish skin gelatin (FSG) and sodium alginate (Alg). The effects of the FSG-Alg complex on the emulsion stability and BITC release pattern from the emulsion were investigated in vitro and in vivo. RESULTS: The storage stability and embedding rate of the 10 g kg-1 FSG and 2.5 g kg-1 Alg (FSG-Alg) emulsion were the highest among all samples. The FSG-Alg complex provided BITC a better protection during in vitro digestion. The microstructure of the FSG-Alg emulsions was more stable during in vitro digestion, and the bioaccessibility and retention rate of BITC were much higher compared to those of the FSG emulsion. The results of the ex vivo everted gut sac of rat intestine study showed that the FSG-Alg emulsion significantly increased the BITC absorption rate in the duodenum. CONCLUSION: The FSG-Alg emulsion delivery system is a highly stable system for the delivery of BITC that improves the bioaccessibility of BITC and promotes its absorption in the duodenum. © 2022 Society of Chemical Industry.

A nanocomposite prepared from silver nanoparticles and carbon dots with peroxidase mimicking activity for colorimetric and SERS-based determination of uric acid.

Silver-carbon dots (Ag-CDs) nanocomposites with excellent peroxidase-like and surface-enhanced Raman scattering (SERS) activities were fabricated by reducing silver ion with carbon dots. The formation of the core-shell structure was demonstrated by transmission electron microscopy. The Ag-CD nanocomposite catalyzes the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2 to form oxidized TMB (oxTMB) that has a blue color with an absorption maximum at 652 nm. The catalytic activity originates from the fact that the electrons of CDs are transferred to H2O2 and decompose H2O2 into hydroxy radicals. The nanocomposites can be used for uric acid (UA) detection because UA can reduce oxTMB to form colorless TMB. The absorbance drops as the concentration of UA increases from 1 to 500 µM. The SERS signal of oxTMB can be detected (at 1605 cm-1) using the Ag-CD nanocomposites as SERS substrate. The intensity of the SERS signal decreases when the concentration of UA ranges from 0.01 to 500 µM. Graphical abstract Schematic representation of the fabrication of silver-carbon dots (Ag-CDs). The Ag-CDs catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) by H2O2 to form blue-colored oxidized TMB (oxTMB). UA reduces oxTMB to form colorless TMB. This process is monitored by surface-enhanced Raman scattering (SERS) spectra for UA detection.

Association of white matter integrity and cognitive functions in patients with subcortical silent lacunar infarcts.

BACKGROUND AND PURPOSE: Silent lacunar infarct (SLI) is a risk factor for dementia. This study investigated the white matter integrity abnormality and its relationship to the cognition impairments in SLI patients. METHODS: Between 27 patients with SLI in basal ganglia and 30 healthy controls, we assessed the difference in a batter of neuropsychological tests and in white matter integrity measurements, fractional anisotropy, and mean diffusivity, using tract-based spatial statistics. RESULTS: Compared with healthy controls, SLI patients performed worse in general mental status, memory, executive function, and language ability. They also had reduced fractional anisotropy and increase mean diffusivity in brain regions such as the body and genu of corpus callosum, the forceps minor, the bilateral superior and bilateral anterior corona radiate, and the left external capsule. Furthermore, we found that in SLI patients, fractional anisotropy measure in left external capsule was positively correlated to the performance in memory and language ability. CONCLUSIONS: SLI in basal ganglia leads to local and remote white matter integrity damages and to the cognition impairments.

An analysis of the influencing factors of false negative autoantibodies in patients with non-small cell lung cancer.

Objectives: To analyze the clinical significance of seven autoantibodies (P53, PGP9.5, SOX2, GAGE7, GBU4-5, MAGE, and CAGE) in patients with non-small cell lung cancer (NSCLC) and the factors that influence false-negative results. Methods: Seven autoantibodies were measured in the serum of 502 patients with non-small cell lung cancer (NSCLC) using ELISA, and their correlations with age, sex, smoking history, pathological type, clinical stage, and PD-L1 gene expression were analyzed. The clinicopathological data of the false-negative and positive groups for the seven autoantibodies were compared to determine the influencing factors. Results: P53 antibody expression level was correlated with lobulation sign, PGP9.5 antibody expression level with sex and vascular convergence; SOX2 antibody expression level with pathological type, clinical stage, and enlarged lymph nodes; and MAGE antibody expression level with the pathological type (P<0.05). False-negative autoantibodies are prone to occur in lung cancer patients with ground-glass nodules, no enlarged lymph nodes, no vascular convergence, and PD-L1 gene expression <1% (P <0.05). Conclusion: Detection of seven autoantibodies was clinically significant in patients with NSCLC. However, poor sensitivity should be considered in clinical diagnoses to prevent missed diagnoses.

Direct synthesis of a lithium carboxymethyl cellulose binder using wood dissolving pulp for high-performance LiFePO4 cathodes in lithium-ion batteries.

Lithium carboxymethyl cellulose (CMC-Li) is a promising novel water-based binder for lithium-ion batteries. The direct synthesis of CMC-Li was innovatively developed using abundant wood dissolving pulp materials from hardwood (HW) and softwood (SW). The resulting CMC-Li-HW and CMC-Li-SW binders possessed a suitable degree of substitutions and excellent molecular weight distributions with an appropriate quantity of long- and short-chain celluloses, which facilitated the construction of a reinforced concrete-like bonding system. When used as cathode binders in LiFePO4 batteries, they uniformly coated and dispersed the electrode materials, formed a compact and stable conductive network with high mechanical strength and showed sufficient lithium replenishment. The prepared LiFePO4 batteries exhibited good mechanical stability, low charge transfer impedance, high initial discharge capacity (∼180 mAh/g), high initial Coulombic efficiency (99 %), excellent cycling performance (<3% loss over 200 cycles) and good rate capability, thereby outperforming CMC-Na and the widely used cathode binder polyvinylidene fluoride.

NADPH oxidase 2 mediates cardiac sympathetic denervation and myocyte autophagy, resulting in cardiac atrophy and dysfunction in doxorubicin-induced cardiomyopathy.

Doxorubicin has been used extensively as a potent anticancer agent, but its clinical use is limited by its cardiotoxicity. However, the underlying mechanisms remain to be fully elucidated. In this study, we tested whether NADPH oxidase 2 (Nox2) mediates cardiac sympathetic nerve terminal abnormalities and myocyte autophagy, resulting in cardiac atrophy and dysfunction in doxorubicin-induced heart failure. Nox2 knockout (KO) and wild-type (WT) mice were randomly assigned to receive a single injection of doxorubicin (15 mg/kg, i.p.) or saline. WT doxorubicin mice exhibited the decreases in survival rate, left ventricular (LV) wall thickness and LV fractional shortening and the increase in the lung wet-to-dry weight ratio 1 week after the injections. These alterations were attenuated in Nox2 KO doxorubicin mice. In WT doxorubicin mice, myocardial oxidative stress was increased, myocardial noradrenergic nerve fibers were reduced, myocardial expression of PGP9.5, GAP43, tyrosine hydroxylase and norepinephrine transporter was decreased, and these changes were prevented in Nox2 KO doxorubicin mice. Myocyte autophagy was increased and myocyte size was decreased in WT doxorubicin mice, but not in Nox2 KO doxorubicin mice. Nox2 mediates cardiac sympathetic nerve terminal abnormalities and myocyte autophagy-both of which contribute to cardiac atrophy and failure after doxorubicin treatment.

Probing the effects of cysteine residues on protein adsorption onto gold nanoparticles using wild-type and mutated GB3 proteins.

The role of cysteine residues in the protein binding kinetics and stability on gold nanoparticles (AuNP) was studied using AuNP localized surface plasmon resonance (LSPR) in combination with an organothiol (OT) displacement method. GB3, the third IgG-binding domain of protein G, was used to model protein-AuNP adsorption. While wild-type GB3 (GB30) contains no cysteine residues, bioengineered GB3 variants containing one (GB31) and two (GB32) cysteine residues were also tested. The cysteine content has no significant effect on GB3 binding kinetics with AuNPs, and most protein adsorption occurs within the first few seconds upon protein/AuNP mixing. However, the stability of GB3 on the AuNP surface against OT displacement depends strongly on the cysteine content and the age of the AuNP/GB3 mixture. The GB30 covered AuNPs can be completely destabilized and aggregated by OTs, regardless of the age of the GB30/AuNP mixtures. Long-time incubation of GB31 or GB32 with AuNPs can stabilize AuNPs against the OT adsorption inducted aggregation. This study indicates that multiple forces involved in the GB3/AuNP interaction, and covalent binding between cysteine and AuNP is essential for a stable protein/AuNP complex.

The relationship with and effect of oral microbiota on NLRP3 inflammatory pathway in type 2 diabetes mellitus.

OBJECTIVE: The aim of this study was to explore the correlation between oral microbiota and NLRP3 inflammasome in type 2 diabetes, and to preliminarily explore their possible impact on type 2 diabetes. DESIGN: The 16S rDNA sequencing technique was used to analyze the microbial composition in the saliva of patients with T2DM and healthy people. Real-time quantitative PCR (RT-qPCR) was used to detect the expression levels of T2DM-related inflammatory cytokines in the blood of two groups. RESULTS: The relative abundances of Fusobacteriota and Campilobacterota in the saliva of patients with T2DM were lower than those of healthy people (P < 0.05), whereas the relative abundance of Proteobacteria in patients with T2DM was higher than that of healthy people (P < 0.05). In addition, real-time quantitative PCR results showed changes in inflammasome-associated factors in the blood of patients with T2DM and healthy people. Compared with healthy individuals, the relative expression levels of lipopolysaccharide (LPS), apoptosis-associated point-like protein (ASC), Caspase-1, Caspase-11, nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3 (NLRP3), and interleukin (IL)-1ß were significantly higher in the blood of patients with T2DM, whereas the expression level of insulin receptor substrate-1 (IRS-1) was reduced (P < 0.05). CONCLUSIONS: Our research suggested that changes in the ratio of oral microbial taxa might increase the expression levels of inflammatory and T2DM-related factors by activating the NLRP3 inflammasome pathway. This discovery indicated the imbalance in oral microbiota might have a certain influence on diabetes by triggering an inflammatory response, and provided a new idea for the relationship between T2DM and oral microbiota.

Clinical-grade human umbilical cord-derived mesenchymal stem cells improved skeletal muscle dysfunction in age-associated sarcopenia mice.

With the expansion of the aging population, age-associated sarcopenia (AAS) has become a severe clinical disease of the elderly and a key challenge for healthy aging. Regrettably, no approved therapies currently exist for treating AAS. In this study, clinical-grade human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) were administrated to two classic mouse models (SAMP8 mice and D-galactose-induced aging mice), and their effects on skeletal muscle mass and function were investigated by behavioral tests, immunostaining, and western blotting. Core data results showed that hUC-MSCs significantly restored skeletal muscle strength and performance in both mouse models via mechanisms including raising the expression of crucial extracellular matrix proteins, activating satellite cells, enhancing autophagy, and impeding cellular aging. For the first time, the study comprehensively evaluates and demonstrates the preclinical efficacy of clinical-grade hUC-MSCs for AAS in two mouse models, which not only provides a novel model for AAS, but also highlights a promising strategy to improve and treat AAS and other age-associated muscle diseases. This study comprehensively evaluates the preclinical efficacy of clinical-grade hUC-MSCs in treating age-associated sarcopenia (AAS), and demonstrates that hUC-MSCs restore skeletal muscle strength and performance in two AAS mouse models via raising the expression of extracellular matrix proteins, activating satellite cells, enhancing autophagy, and impeding cellular aging, which highlights a promising strategy for AAS and other age-associated muscle diseases.

An optimized method for obtaining clinical-grade specific cell subpopulations from human umbilical cord-derived mesenchymal stem cells.

Mesenchymal stem cells (MSCs) are heterogeneous populations with broad application prospects in cell therapy, and using specific subpopulations of MSCs can enhance their particular capability under certain conditions and achieve better therapeutic effects. However, no studies have reported how to obtain high-quality specific MSC subpopulations in vitro culture. Here, for the first time, we established a general operation process for obtaining high-quality clinical-grade cell subpopulations from human umbilical cord MSCs (hUC-MSCs) based on particular markers. We used the MSC-CD106+ subpopulations, whose biological function has been well documented, as an example to explore and optimize the crucial links of primary preparation, pre-treatment, antibody incubation, flow sorting, quality and function test. After comprehensively evaluating the quality and function of the acquired MSC-CD106+ subpopulations, including in vitro cell viability, apoptosis, proliferation, marker stability, adhesion ability, migration ability, tubule formation ability, immunomodulatory function and in vivo wound healing ability and proangiogenic activity, we defined an important pre-treatment scheme which might effectively improve the therapeutic efficiency of MSC-CD106+ subpopulations in two critical clinical application scenarios-direct injection after cell sorting and post-culture injection into bodies. Based on the above, we tried to establish a general five-step operation procedure for acquiring high-quality clinical-grade MSC subpopulations based on specific markers, which cannot only improve their enrichment efficiency and the reliability of preclinical studies, but also provide valuable methodological guidance for the rapid clinical transformation of specific MSC subpopulations.

Gegen Qinlian Decoction ameliorates type 2 diabetes osteoporosis via IGFBP3/MAPK/NFATc1 signaling pathway based on cytokine antibody array.

BACKGROUND: Osteoporosis affects more than half the patients with type 2 diabetes mellitus (T2DM). Up to data, there is no effective clinical practice in managing type 2 diabetes osteoporosis (T2DOP) because of its complex pathogenesis. Gegen Qinlian Decoction (GQD) has been used for the long-term management of T2DM. However, the underlying mechanism of GQD in the treatment of T2DOP remains unknown. PURPOSE: To reveal the role of GQD in T2DOP and its potential therapeutic targets in the management of T2DOP. STUDY DESIGN: The effect of GQD on T2DOP was observed in db/db mice in four groups: model group, GQD low-dose group (GQD-L), GQD high-dose group (GQD-H), and metformin (positive control) group. C57BL/6J mice were used as the negative control group. METHODS: Quantitative phytochemical analysis of GQD was performed using high-performance liquid chromatography (HPLC). Micro-CT and hematoxylin-eosin (H&E) staining were used to evaluate bone histomorphometry. To screen for candidate targets of GQD, a cytokine antibody array was used, followed by bioinformatics analysis. Quantitative real-time PCR (qRT-PCR) and western blotting (WB) were used to determine expression levels. RESULTS: The major active components of GQD were confirmed by HPLC. Micro-CT and H&E staining showed that bone mass was significantly increased in the GQD-H group compared with the model group. Antibody arrays revealed that the expression of insulin-like growth factor binding protein 3 (IGFBP3) was elevated in the GQD-H group. The MAPK pathway was identified using bioinformatics analysis. Additionally, the levels of osteoclastogenesis-related genes, including cathepsin K (Ctsk), acid phosphatase 5 (Acp5), matrix metallopeptidase 9 (Mmp9), and ATPase H+ transporting V0 subunit D2 (Atp6v0d2) were significantly decreased in the GQD-H group. Compared with the model group, high-dosage GQD inhibited phosphorylation of extracellular signal-regulated kinases (ERKs) and P38 mitogen-activated protein kinase (MAPK) and the expression of c-Fos and nuclear factor of activated T cells 1 (NFATc1). CONCLUSION: GQD plays a protective role in T2DOP by upregulating IGFBP3 expression and downregulating the IGFBP3/MAPK/NFATc1 signaling pathway. IGFBP3 in serum may also be a novel biomarker in the treatment of T2DOP. Our current findings not only expand the application of GQD, but also provide a theoretical basis and guidance for T2DOP.

Broad ultra-potent neutralization of SARS-CoV-2 variants by monoclonal antibodies specific to the tip of RBD.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) continue to wreak havoc across the globe. Higher transmissibility and immunologic resistance of VOCs bring unprecedented challenges to epidemic extinguishment. Here we describe a monoclonal antibody, 2G1, that neutralizes all current VOCs and has surprising tolerance to mutations adjacent to or within its interaction epitope. Cryo-electron microscopy structure showed that 2G1 bound to the tip of receptor binding domain (RBD) of spike protein with small contact interface but strong hydrophobic effect, which resulted in nanomolar to sub-nanomolar affinities to spike proteins. The epitope of 2G1 on RBD partially overlaps with angiotensin converting enzyme 2 (ACE2) interface, which enables 2G1 to block interaction between RBD and ACE2. The narrow binding epitope but high affinity bestow outstanding therapeutic efficacy upon 2G1 that neutralized VOCs with sub-nanomolar half maximal inhibitory concentration in vitro. In SARS-CoV-2, Beta or Delta variant-challenged transgenic mice and rhesus macaque models, 2G1 protected animals from clinical illness and eliminated viral burden, without serious impact to animal safety. Mutagenesis experiments suggest that 2G1 is potentially capable of dealing with emerging SARS-CoV-2 variants in the future. This report characterized the therapeutic antibodies specific to the tip of spike against SARS-CoV-2 variants and highlights the potential clinical applications as well as for developing vaccine and cocktail therapy.