Polyoxometalate-based nanozyme with laccase-mimicking activity for kanamycin detection based on colorimetric assay.

As a kind of aminoglycoside antibiotics, kanamycin (KAN) is widely applied to animal husbandry and aquaculture. However, the abuse of KAN causes the large-scale discharge of it into rivers, lakes and groundwater, which threatens environmental safety and human health. Therefore, it is imperative to develop a method that is applicable to detect KAN in an efficient and accurate way. The colorimetric method based on enzymes provides a feasible solution for the detection of organic pollutants. However, the extensive application of natural enzymes is constrained by high cost and low stability. Herein, a polyoxometalate-based nanozyme, namely [H7SiW9V3O40(DPA)3]·4H2O (SiW9V3/DPA) (DPA = dipyridylamine), is synthesized. As a low-cost nanozyme with high stability compared to natural enzymes, SiW9V3/DPA performs well in laccase-mimicking activity. It can be used to induce chromogenic reaction between 2,4-dichlorophenol (2,4-DP) and 4-aminoantipyrine (4-AP), which generates red products. With the addition of KAN, the color fades. That is to say, KAN can be detected with colorimetric assay in the concentration range 0.1 to 100 µM with high selectivity and low limit of detection (LOD) of 6.28 µM. Moreover, SiW9V3/DPA is applied to KAN detection in lake and river water and milk with satisfactory results. To sum up, polyoxometalate-based nanozyme is expected to provide a promising solution to the detection of organic pollutants in the aquatic environment.

Targeting a cytokine checkpoint enhances the fitness of armored cord blood CAR-NK cells.

Immune checkpoint therapy has resulted in remarkable improvements in the outcome for certain cancers. To broaden the clinical impact of checkpoint targeting, we devised a strategy that couples targeting of the cytokine-inducible Src homology 2-containing (CIS) protein, a key negative regulator of interleukin 15 (IL-15) signaling, with fourth-generation "armored" chimeric antigen receptor (CAR) engineering of cord blood-derived natural killer (NK) cells. This combined strategy boosted NK cell effector function through enhancing the Akt/mTORC1 axis and c-MYC signaling, resulting in increased aerobic glycolysis. When tested in a lymphoma mouse model, this combined approach improved NK cell antitumor activity more than either alteration alone, eradicating lymphoma xenografts without signs of any measurable toxicity. We conclude that targeting a cytokine checkpoint further enhances the antitumor activity of IL-15-secreting armored CAR-NK cells by promoting their metabolic fitness and antitumor activity. This combined approach represents a promising milestone in the development of the next generation of NK cells for cancer immunotherapy.

Knowledge, Attitude, and Practice of Breast Cancer Patients Toward Lymphedema Complications: Cross-Sectional Study.

Breast cancer is commonly treated through surgical resection, but a common complication of the procedure is lymphedema of the upper limbs, which can significantly impact patients' daily life. This study aims to investigate the knowledge, attitude, and practice (KAP) of breast cancer patients with regard to lymphedema complications. This cross-sectional study was conducted by a self-administered questionnaire between August and October 2022 toward breast cancer patients in our Hospital of Traditional Chinese Medicine. A total of 529 breast cancer patients were enrolled, including 186 (35.16%) aged < 50 years old. Participants had moderate knowledge, attitudes, and practices with scores of 18.24 ± 3.145 (possible range: 0-30), 62.24 ± 10.260 (possible range: 17-85), and 63.27 ± 20.967 (possible range: 21-105), respectively. Multivariate logistic regression showed that high school/technical secondary school (OR = 1.880, 95% CI = 1.107-3.194, P = 0.019) and being retired (OR = 0.482, 95% CI = 0.245-0.947, P = 0.034) were independently associated with good knowledge. Knowledge (OR = 1.321, 95% CI = 1.222-1.428, P < 0.001) was independently associated with a good attitude. Furthermore, knowledge (OR = 1.262, 95% CI = 1.151-1.384, P < 0.001) and attitude (OR = 1.122, 95% CI = 1.085-1.160, P < 0.001) were independently associated with good practice. Breast cancer patients have moderate knowledge, attitudes, and practices regarding lymphedema complications. Effective education and self-management programs are needed to improve patients' KAP toward lymphedema.

Non-fucosylated CB CD34+ cells represent a good target for enforced fucosylation to improve engraftment following cord blood transplantation.

BACKGROUND AIMS: Despite ethnic diversity and ready availability of cryopreserved, human leukocyte antigen-typed cord blood (CB), delayed engraftment remains a significant hurdle to successful CB transplantation. Suboptimal homing of CB hematopoietic stem and progenitor cells (HSPCs) to the hematopoietic microenvironment (HM) is thought to be responsible and due to low levels of HSPC fucosylation. Fucosylation (decoration with sialyl-LewisX) may improve HSPC homing to HM by increasing the strength of HSPC/E-selectin interactions, where E-selectin is constitutively expressed by HM microvasculature. Enforced fucosylation of CB HSPCs using fucosyltransferases, increases the rate and magnitude of engraftment in xenogeneic transplant models. However, it is unclear whether endogenously fucosylated and non-fucosylated CB HSPC are qualitatively identical or whether endogenous fucosylation marks a qualitative difference between CB HSPC. If qualitatively identical, non-fucosylated CB HSPCs represent a good target for enforced fucosylation with improved engraftment conferred on an increased number of otherwise qualitatively identical HSPC. If qualitatively different, then conferring engraftment upon a majority, possibly lower "quality," non-fucosylated HSPCs by enforced fucosylation might inadvertently compromise engraftment. METHODS: Functional (xenogeneic engraftment, colony-forming unit and selectin-binding assays) and phenotypic analyses of fluorescence-activated cell sorting-isolated, endogenously fucosylated and non-fucosylated CB CD34+ cells were performed. RESULTS: Endogenous fucosylation of CB HSPCs exists as a continuum. Endogenously fucosylated HSPCs engrafted more efficiently in a xenogeneic transplantation model than non-fucosylated HSPCs. Outside of the differences in endogenous fucosylation, no other qualitative (functional and/or phenotypic) differences were identified. DISCUSSION: The majority of endogenously non-fucosylated CB HSPCs represent a good target for enforced fucosylation with the goal of improving engraftment following CB transplantation.

Fucosylation with fucosyltransferase VI or fucosyltransferase VII improves cord blood engraftment.

BACKGROUND AIMS: Advantages associated with the use of cord blood (CB) transplantation include the availability of cryopreserved units, ethnic diversity and lower incidence of graft-versus-host disease compared with bone marrow or mobilized peripheral blood. However, poor engraftment remains a major obstacle. We and others have found that ex vivo fucosylation can enhance engraftment in murine models, and now ex vivo treatment of CB with fucosyltransferase (FT) VI before transplantation is under clinical evaluation (NCT01471067). However, FTVII appears to be more relevant to hematopoietic cells and may alter acceptor substrate diversity. The present study compared the ability of FTVI and FTVII to improve the rapidity, magnitude, multi-lineage and multi-tissue engraftment of human CB hematopoietic stem and progenitor cells (HSPCs) in vivo. METHODS: CD34-selected CB HSPCs were treated with recombinant FTVI, FTVII or mock control and then injected into immunodeficient mice and monitored for multi-lineage and multi-tissue engraftment. RESULTS: Both FTVI and FTVII fucosylated CB CD34⁺ cells in vitro, and both led to enhanced rates and magnitudes of engraftment compared with untreated CB CD34⁺ cells in vivo. Engraftment after treatment with either FT was robust at multiple time points and in multiple tissues with similar multi-lineage potential. In contrast, only FTVII was able to fucosylate T and B lymphocytes. CONCLUSIONS: Although FTVI and FTVII were found to be similarly able to fucosylate and enhance the engraftment of CB CD34⁺ cells, differences in their ability to fucosylate lymphocytes may modulate graft-versus-tumor or graft-versus-host effects and may allow further optimization of CB transplantation.

A Bivalent Aptamer-Based DNA Agonist for EGFR Signaling Effectively Alleviates Ulcerative Colitis In Vivo.

While epidermal growth factor (EGF) shows promise in addressing the clinical manifestations of intestinal ulcerative diseases by activating the EGF receptor (EGFR)-mediated cell signaling, its clinical application is hampered by poor protein hydrolytic stability, low thermostability, and difficulty in modification. The development of a novel EGFR agonist for ulcerative colitis remains an urgent need, necessitating innovative solutions to overcome the limitations of current therapies via recombinant EGF protein. Herein, we introduce a novel DNA agonist for EGFR, Dimer-YL, which employs a bivalent aptamer to induce stable receptor dimerization, thereby activating the EGFR signaling and related cell behaviors. Dimer-YL has been demonstrated to recapitulate the EGF-promoted cellular behaviors, including proliferation and migration, as well as repair the damage of intercellular tight junctions. Furthermore, our findings demonstrate the potent therapeutic function of Dimer-YL in alleviating DSS-induced ulcerative colitis in vivo. Together, the present work has revealed Dimer-YL as an innovative DNA molecule for effective EGFR activation, offering promise for the development of EGFR-agonistic agents for therapeutic purposes.

Association between Timing of labor Induction and Neonatal and Maternal outcomes: An Observational Study from China.

BACKGROUND: Growing evidence suggests that elective induction of labor at 39 weeks may lead to more favorable perinatal outcomes compared with the expectant management, however, how to weigh the pros and cons of elective labor induction at 39 weeks, the expectation of spontaneous delivery at 40 or 41 weeks, or delayed labor induction at 40 or 41 weeks on neonatal and maternal outcomes remains a practical challenge in clinical decision-making. OBJECTIVE: We compared neonatal and maternal outcomes between elective induction of labor at 39 weeks and expectant management in a real word setting. We also divided the expectantly managed group and compared outcomes between the spontaneous delivery group at 40 or 41 weeks, and the induced group at 40 or 41 weeks versus the elective induced group at 39 weeks. STUDY DESIGN: This retrospective cohort study included 21282 participants between January 1, 2019, and June 30, 2022. Participants were initially categorized into three groups at 39 weeks: elective induction of labor, spontaneous delivery, and expectant management, for the primary analysis comparing elective induction with expectant management. Subsequently, the expectant management group at 39 weeks was similarly divided into three groups at 40 weeks, and participants who underwent expectant management at 40 weeks were then divided into two groups at 41 weeks: elective induction and spontaneous delivery. In total, six groups were compared in the secondary analysis, with elective induction at 39 weeks serving as the reference group. RESULTS: At 39 weeks' gestational age, participants who received elective induction of labor had a significantly lower risk of primary composite outcomes compared to participants who received expectant management (adjusted odds ratio [aOR]: 0.72, 95% confidence interval [CI]: 0.55-0.95), and there was no significant difference in risk of cesarean delivery between the two groups. After further dividing the expectantly managed group, compared to participants with elective induction of labor at 39 weeks, those with spontaneous delivery at 40 weeks had significant lower risks of cesarean delivery (0.61, 0.52-0.71) and chorioamnionitis (0.78, 0.61-1.00), but a higher risk of fetal distress (1.39, 1.22-1.57); those with spontaneous delivery at 41 weeks had a significant higher risk of fetal distress (1.44, 1.16-1.79), postpartum hemorrhage (1.83, 1.26-2.66), and prolonged/arrested labor (1.61, 1.02-2.54). Moreover, compared to participants with elective induction of labor at 39 weeks, participants induced at later weeks had significantly higher risks of neonatal and maternal outcomes, especially at 40 weeks. CONCLUSIONS: Our findings indicate that elective induction of labor at 39 weeks was significantly associated with lower risks of short-term neonatal and maternal outcomes compared to expectant management. Moreover, our study highlights the nuanced trade-offs in risks and benefits between elective induction at 39 weeks versus waiting for spontaneous labor or delayed induction at 40/41 weeks, thus providing valuable insights for clinical decision-making in practice.

Effects of different anesthesia methods on postoperative immune function in patients undergoing gastrointestinal tumor resection.

To investigate the effects of different anesthetic methods on postoperative immune function in patients undergoing gastrointestinal tumor resection. Ninety patients undergoing laparoscopic gastrointestinal tumor resection were divided into 3 groups. Patients in the GA group were anesthetized by total intravenous anesthesia. The GE group was anesthetized by general anesthesia combined with epidural anesthesia. The GN group was anesthetized by general anesthesia combined with bilateral Transversus Abdominis Plane block (TAP) and rectus sheath nerve blocks. General anesthesia is total intravenous anesthesia in all three groups. Blood samples were taken to test the changes of peripheral lymphocyte subtype analysis, and levels of plasma cortisol, epinephrine, norepinephrine. Also, the dosage of anesthetic drugs, recovery time, and visual analog scale (VAS) scores were recorded. Postoperative immune indexes, including CD4 count, CD8 count, B, and NK cells, in the GE group were significantly higher than those in NA and GA groups (P < 0.01). Perioperative stress indices, including epinephrine levels, norepinephrine level and aldosterone level, in the GE group were significantly lower than in the GA group and GN group (P < 0.01). The intraoperative/total sufentanil dosage and remifentanil dosage in the GE group were significantly lower than those in the GA and GN groups (P < 0.01). The VAS scores in the GE group were significantly better than those in GA and GN groups (P < 0.01). General anesthesia combined with epidural anesthesia attenuates the increase in inflammatory mediators. Its possible mechanisms include reducing perioperative stress response and reducing perioperative opioid use.

A new three-dimensional zinc(II) metal-organic framework as a fluorescence sensor for sensing the biomarker 3-nitrotyrosine.

3-Nitrotyrosine (3-NT), an oxidative stress biomarker, is closely associated with various diseases. Thus, rapid and sensitive detection of 3-NT is of great significance for preventing and treating diseases. Herein, we reported a new 3D zinc-based metal-organic framework (Zn-MOF) [Zn(L)(HBTC)] (L = (E)-4,4'-(ethene-1,2-diyl)bis[(N-pyridin-3-yl)benzamide], H3BTC = 1,3,5-benzenetricarboxylic acid), which was structurally characterized by single crystal X-ray diffraction, IR, PXRD and TG. The Zn-MOF can be used as a highly efficient fluorescence sensing material to provide a direct and low-cost method for the rapid detection of 3-NT and shows high sensitivity with a KSV value of 6.596 × 104 M-1, a rapid luminescence response within 24 s, excellent selectivity, high anti-interference ability and good recyclability. It is the first example of a MOF being used to directly detect 3-NT as a luminescence sensor to our knowledge. The sensing mechanism of the Zn-MOF towards 3-NT is discussed in detail, which provides a basis for the rational design of MOF sensing materials and their application in biomarker detection.

Activated B cells suppress T-cell function through metabolic competition.

BACKGROUND: B cells play a pivotal role in regulating the immune response. The induction of B cell-mediated immunosuppressive function requires B cell activating signals. However, the mechanisms by which activated B cells mediate T-cell suppression are not fully understood. METHODS: We investigated the potential contribution of metabolic activity of activated B cells to T-cell suppression by performing in vitro experiments and by analyzing clinical samples using mass cytometry and single-cell RNA sequencing. RESULTS: Here we show that following activation, B cells acquire an immunoregulatory phenotype and promote T-cell suppression by metabolic competition. Activated B cells induced hypoxia in T cells in a cell-cell contact dependent manner by consuming more oxygen via an increase in their oxidative phosphorylation (OXPHOS). Moreover, activated B cells deprived T cells of glucose and produced lactic acid through their high glycolytic activity. Activated B cells thus inhibited the mammalian target of rapamycin pathway in T cells, resulting in suppression of T-cell cytokine production and proliferation. Finally, we confirmed the presence of tumor-associated B cells with high glycolytic and OXPHOS activities in patients with melanoma, associated with poor response to immune checkpoint blockade therapy. CONCLUSIONS: We have revealed for the first time the immunomodulatory effects of the metabolic activity of activated B cells and their possible role in suppressing antitumor T-cell responses. These findings add novel insights into immunometabolism and have important implications for cancer immunotherapy.

A metabonomic approach to analyze the dexamethasone-induced cleft palate in mice.

Mice models are an important way to understand the relation between the fetus with cleft palate and changes of maternal biofluid. This paper aims to develop a metabonomics approach to analyze dexamethasone-induced cleft palate in pregnant C57BL/6J mice and to study the relationship between the change of endogenous small molecular metabolites in maternal plasma and the incidence of cleft palate. To do so, pregnant mice were randomly divided into two groups. The one group was injected with dexamethasone. On E17.5th day, the incident rates of cleft palate from embryos in two groups were calculated. The (1)H-NMR spectra from the metabolites in plasma in two groups was collected at same time. Then the data were analyzed using metabonomics methods (PCA and SIMCA). The results showed that the data from the two groups displayed distinctive characters, and the incidence of cleft palate were significantly different (P < .005). To conclude, this study demonstrates that the metabonomics approach is a powerful and effective method in detecting the abnormal metabolites from mother in the earlier period of embryos, and supports the idea that a change from dexamethasone induced in maternal metabolites plays an important role in the incidence of cleft palate.

Polymerase chain reaction detection of Lactobacillus acidophilus in human oral cavity and fecal samples after 2-week consumption of yoghurt.

OBJECTIVE: To investigate whether short-term daily consumption of yoghurt leads to colonization by Lactobacillus acidophilus in a group of human subjects who were initially totally devoid of L. acidophilus in their oral cavities. MATERIAL AND METHODS: Twenty-three volunteers consumed yogurt containing L. acidophilus during a 14-day trial stage. Oral and fecal samples were collected at the clearance stage and at the post-yoghurt intake stage until L. acidophilus was found. Standard polymerase chain reaction methods using specific primers were adopted for the detection and identification of L. acidophilus. RESULTS: The isolation frequency decreased rapidly 72 h after stopping intake of yoghurt. After 1 week, L. acidophilus was absent in all oral samples. Non-significant differences were found between the survival rates of L. acidophilus in samples of saliva, plaque, tongue surface, and buccal mucosa. L. acidophilus was also found to remain in the gastrointestinal tract for longer than in the oral cavity. CONCLUSION: Allochthonous L. acidophilus is not likely to permanently colonize the oral cavity and intestine.

Two new polyoxometalate-based metal-organic complexes for the detection of trace Cr(VI) and their capacitor performance.

Two new Keggin-type polyoxometalate (POM)-based metal-organic complexes (MOCs) H3[Cu2(4-dpye)2(PMo12O40)] (1) and H[Cu2(4-Hdpye)2(PMo12O40)(H2O)4]·2H2O (2) were constructed with a new N,N'-bis (4-pyrimidinecarboxamido)-1,2-ethane (4-H2dpye) ligand by the hydrothermal/solvothermal method. Complex 1 was a 2D layered structure constructed from 1D metal-organic chains [Cu(4-dpye)]n and Keggin-type [PMo12O40]3- polyoxoanions. Complex 2 displays a 3D supramolecular framework formed by discrete [PMo12O40]3- polyoxoanions and binuclear metal-organic loops [Cu2(4-Hdpye)2]. The electrocatalytic behaviors of carbon paste electrodes modified by complexes 1 and 2 (1-CPE and 2-CPE) were investigated. The 1-CPE and 2-CPE were used as electrochemical sensors to detect trace Cr(vi), and the low limits of detection (LOD) are 1.27 × 10-7 M for 1 and 1.71 × 10-7 M for 2, which are lower than the maximum allowable concentration of Cr(vi) in drinking water specified by the World Health Organization (WHO). In addition, the performances of complexes 1 and 2 modified carbon cloth electrodes (1-CC and 2-CC) as supercapacitor materials have also been studied. The influence of the structure on electrocatalytic and capacitor performances is discussed.

Metabolic Reprogramming of GMP Grade Cord Tissue Derived Mesenchymal Stem Cells Enhances Their Suppressive Potential in GVHD.

Acute graft-vs.-host (GVHD) disease remains a common complication of allogeneic stem cell transplantation with very poor outcomes once the disease becomes steroid refractory. Mesenchymal stem cells (MSCs) represent a promising therapeutic approach for the treatment of GVHD, but so far this strategy has had equivocal clinical efficacy. Therapies using MSCs require optimization taking advantage of the plasticity of these cells in response to different microenvironments. In this study, we aimed to optimize cord blood tissue derived MSCs (CBti MSCs) by priming them using a regimen of inflammatory cytokines. This approach led to their metabolic reprogramming with enhancement of their glycolytic capacity. Metabolically reprogrammed CBti MSCs displayed a boosted immunosuppressive potential, with superior immunomodulatory and homing properties, even after cryopreservation and thawing. Mechanistically, primed CBti MSCs significantly interfered with glycolytic switching and mTOR signaling in T cells, suppressing T cell proliferation and ensuing polarizing toward T regulatory cells. Based on these data, we generated a Good Manufacturing Process (GMP) Laboratory protocol for the production and cryopreservation of primed CBti MSCs for clinical use. Following thawing, these cryopreserved GMP-compliant primed CBti MSCs significantly improved outcomes in a xenogenic mouse model of GVHD. Our data support the concept that metabolic profiling of MSCs can be used as a surrogate for their suppressive potential in conjunction with conventional functional methods to support their therapeutic use in GVHD or other autoimmune disorders.

Targeting the αv integrin/TGF-ß axis improves natural killer cell function against glioblastoma stem cells.

Glioblastoma multiforme (GBM), the most aggressive brain cancer, recurs because glioblastoma stem cells (GSCs) are resistant to all standard therapies. We showed that GSCs, but not normal astrocytes, are sensitive to lysis by healthy allogeneic natural killer (NK) cells in vitro. Mass cytometry and single-cell RNA sequencing of primary tumor samples revealed that GBM tumor-infiltrating NK cells acquired an altered phenotype associated with impaired lytic function relative to matched peripheral blood NK cells from patients with GBM or healthy donors. We attributed this immune evasion tactic to direct cell-to-cell contact between GSCs and NK cells via αv integrin-mediated TGF-ß activation. Treatment of GSC-engrafted mice with allogeneic NK cells in combination with inhibitors of integrin or TGF-ß signaling or with TGFBR2 gene-edited allogeneic NK cells prevented GSC-induced NK cell dysfunction and tumor growth. These findings reveal an important mechanism of NK cell immune evasion by GSCs and suggest the αv integrin/TGF-ß axis as a potentially useful therapeutic target in GBM.

An electron microscopy study of the diversity of Streptococcus sanguinis cells induced by lysozyme in vitro.

Bacterial virulence could be altered by the antimicrobial agents of the host. Our aim was to identify the damage and survival of Streptococcus sanguinis induced by lysozymes in vitro and to analyse the potential of oral microorganisms to shirk host defences, which cause infective endocarditis. S. sanguinis ATCC 10556 received lysozyme at concentrations of 12.5, 25, 50 and 100 microg/ml. Cells were examined by electron microscopy. The survival was assessed by colony counting and construction of a growth curve. Challenged by lysozymes, cells mainly exhibited cell wall damage, which seemed to increase with increasing lysozyme concentration and longer incubation period in the presence of ions. Cells with little as well as apparent lesion were observed under the same treatment set, and anomalous stick and huge rotund bodies were occasionally observed. After the removal of the lysozyme, some damaged cells could be reverted to its original form with brain heart infusion (BHI), and their growth curve was similar to the control cells. After further incubation in BHI containing lysozyme, S. sanguinis cell damage stopped progressing, and their growth curve was also similar to the control cells. The results suggested that the S. sanguinis lesions caused by the lysozyme in the oral cavity may be nonhomogeneous and that some damaged cells could self-repair and survive. It also indicated that S. sanguinis with damaged cell walls may survive and be transmitted in the bloodstream.

The absorption characteristics of bifendate solid dispersion in rat intestinal tissue.

AIM: Research on bifendate intestinal absorption. METHOD: The single passed intestinal backflow method was used. Bifendate with different concentrations, bifendate with and without 2,4-dinitrophenol, verapamil, and probenecid, and solid state of bifendate in different systems were studied. RESULT: Change of concentration and the presence of energy inhibitor, P-glycoprotein inhibitor and multidrug-resistant protein inhibitor did not affect the K(a) of bifendate intestinal absorption; there were obvious differences among intestinal absorption of native drug, solid dispersion, and physical mixtures. CONCLUSION: The result indicated that the intestinal absorption mechanism of bifendate is passive transport. Solid state of bifendate in different systems could affect the intestinal absorption.

Effect of toothpaste containing d-limonene on natural extrinsic smoking stain: a 4-week clinical trial.

PURPOSE: To determine whether natural smoking stain could be removed/inhibited effectively by a toothpaste containing 5% d-limonene. For comparison and contrast, the effects of d-limonene on tea stain were also assessed. METHODS: The design was a randomized controlled double-blind trial with parallel groups. Toothpastes were: A: positive control with perlite whitening formulation; B: A+5% d-limonene; C: D + 5% d-limonene; D: negative control. The extrinsic stains were measured using Lobene Stain Index. Following baseline examination, all subjects were randomly assigned to one of the four toothpaste groups and instructed to brush with the assigned products twice daily. Subjects returned to the clinic after 4-week brushing for stain removal assessment, then all extrinsic stains, plaque and supragingival calculus were removed and use of assigned products was continued for another 4 weeks, and the stain scores were repeated for inhibition assessment. RESULTS: A total of 408 subjects, 201 with smoking stains and 207 with tea stains, participated in the trial. 5% d-limonene combined with Perlite whitening formulation significantly reduced stain scores both for smoking stain removal and inhibition (P < 0.05). Furthermore, 5% d-limonene alone (in negative formulation) exhibited an additional advantage for smoking stain inhibition (P < 0.05), but the advantage was not found for long-standing smoking stain removal (P > 0.05). The additional advantage of 5% d-limonene was shown neither for removal nor for inhibition in the tea stain study (P > 0.05). All test products were well tolerated over the study period.

Large-scale GMP-compliant CRISPR-Cas9-mediated deletion of the glucocorticoid receptor in multivirus-specific T cells.

Virus-specific T cells have proven highly effective for the treatment of severe and drug-refractory infections after hematopoietic stem cell transplant (HSCT). However, the efficacy of these cells is hindered by the use of glucocorticoids, often given to patients for the management of complications such as graft-versus-host disease. To address this limitation, we have developed a novel strategy for the rapid generation of good manufacturing practice (GMP)-grade glucocorticoid-resistant multivirus-specific T cells (VSTs) using clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) gene-editing technology. We have shown that deleting the nuclear receptor subfamily 3 group C member 1 (NR3C1; the gene encoding for the glucocorticoid receptor) renders VSTs resistant to the lymphocytotoxic effect of glucocorticoids. NR3C1-knockout (KO) VSTs kill their targets and proliferate successfully in the presence of high doses of dexamethasone both in vitro and in vivo. Moreover, we developed a protocol for the rapid generation of GMP-grade NR3C1 KO VSTs with high on-target activity and minimal off-target editing. These genetically engineered VSTs promise to be a novel approach for the treatment of patients with life-threatening viral infections post-HSCT on glucocorticoid therapy.

[The effect of polyamidoamine (PAMAM) dendrimers on the solubility and pharmacokinetics of breviscapine].

To study the solubilization of breviscapine with polyamidoamine (PAMAM) dendrimers and probe the solubilizing mechanism and investigate the influence of PAMAM dendrimers on the pharmacokinetics of breviscapine, the solubilization of breviscapine by PAMAM dendrimers of generations G1, G1.5, G2 and G2.5 with different concentrations were determined and compared in different pH conditions. Twelve rats randomized into 2 groups were separately orally administered breviscapine and breviscapine combining with PAMAM. Drug in plasma was extracted and determined with HPLC. In pH condition lower than 7.0, the solubilization of breviscapine by PAMAM dendrimers enhanced as the generation and concentration of PAMAM dendrimers as well as the pH increased. Its solubilizing mechanism involves an electrostatic interaction between the carboxyl group of breviscapine and the primary amines and tertiary amines of PAMAM dendrimers. The pharmacokinetics parameters Cmax and AUC0-8 h of breviscapine were (119.65 +/- 9.36) ng x mL(-1) and (370.09 +/- 63.08) ng x h x mL(-1). For breviscapine combined with PAMAM dendrimers, the Cmax and AUC0-8 h were (518.17 +/- 17.07) ng x mL(-1) and (1,219.47 +/- 201.87) ng x h x mL(-1), respectively. There were significant differences of AUC0-8 h between breviscapine and breviscapine combined with PAMAM dendrimers (P < 0.01). PAMAM dendrimers can greatly increase the solubility of breviscapine in water and can improve the oral bioavailability of breviscapine significantly.