Kinetics of sterilization of atomized slightly acidic electrolyzed water on tableware.

The aim of this study was to elucidate the kinetics of atomization of slightly acidic electrolyzed water (SAEW) for use in sterilization of secondary contaminated tableware surfaces. The sterilization efficacy of SAEW was assessed on the basis of the change in the total number of colonies with different contamination levels (101 CFU/mL and 102 CFU/mL), atomization time (10, 20, 30, 40, and 50 s), atomizing distance (5, 10, 15, 20, 25, and 30 cm), and available chlorine concentration (ACC; 25.2, 30.2, 34.9, 40.5, 44.8, and 53.3 mg/L) as the main influencing factors. According to the relationship among flux, atomization area, and time, a kinetic model of SAEW atomization for the sterilization of tableware surfaces was established. The results indicated that the sterilization efficacy of SAEW gradually improved with decreased contamination levels (12.69 %-15.74 %), extended atomization time (13.68 %-46.58 %), and increased ACC (36.89 %-95.14 %). Based on the kinetics analysis, the change law of the kinetic model of SAEW atomization and sterilization of tableware surfaces with secondary pollution was found to be consistent with the change law of sterilization (r2 > 0.8). The results of this study provide a theoretical basis for SAEW atomization for sterilization of secondary contaminated tableware surfaces and also contributes to the improvement of technological theory of SAEW sterilization.

PA suppresses antitumor immunity of T cells by disturbing mitochondrial activity through Akt/mTOR-mediated Ca2+ flux.

Deciphering the mechanisms behind how T cells become exhausted and regulatory T cells (Tregs) differentiate in a tumor microenvironment (TME) will significantly benefit cancer immunotherapy. A common metabolic alteration feature in TME is lipid accumulation, associated with T cell exhaustion and Treg differentiation. However, the regulatory role of free fatty acids (FFA) on T cell antitumor immunity has yet to be clearly illustrated. Our study observed that palmitic acid (PA), the most abundant saturated FFA in mouse plasma, enhanced T cell exhaustion and Tregs population in TME and increased tumor growth. In contrast, oleic acid (OA), a monounsaturated FFA, rescued PA-induced T cell exhaustion, decreased Treg population, and ameliorated T cell antitumor immunity in an obese mouse model. Mechanistically, mitochondrial metabolic activity is critical in maintaining T cell function, which PA attenuated. PA-induced T cell exhaustion and Treg formation depended on CD36 and Akt/mTOR-mediated calcium signaling. The study described a new mechanism of PA-induced downregulation of antitumor immunity of T cells and the therapeutic potential behind its restoration by targeting PA.

Herpes Virus Entry Mediator Costimulation Signaling Enhances CAR T-cell Efficacy Against Solid Tumors Through Metabolic Reprogramming.

Costimulatory domains (CSD) of 4-1BB and CD28 are most widely used in chimeric antigen receptor (CAR)-engineered T cells. These CAR T cells have shown encouraging efficacy in the treatment of hematologic malignancies but have limited efficacy in solid tumors. The herpes virus entry mediator (HVEM) is a costimulatory molecule with a novel downstream signaling pathway. In response to target cells, CAR T cells with a HVEM CSD (HVEM-CAR T) displayed more robust cytokine release and cytotoxicity than 4-1BB-CAR T or CD28-CAR T in vitro. Furthermore, HVEM-CAR T showed superior therapeutic efficacy in several mouse tumor models. Mechanistically, the HVEM CSD endowed CAR T cells with attenuated exhaustion, improved function and persistence, and enhanced metabolic activities in tumor tissue compared with 4-1BB-based or CD28-based CAR T cells. These studies establish that the HVEM CSD has the potential to improve the therapeutic efficacy of CAR T cells against solid tumors.

Docetaxel enhances the therapeutic efficacy of PSMA-specific CAR-T cells against prostate cancer models by suppressing MDSCs.

PURPOSE: Prostate cancer can undergo curative effects by radical prostatectomy or radical radiotherapy. However, the best treatment for more aggressive high-risk prostate cancer remains controversial. Insufficient infiltration capacity and dysfunction are commonly occurrences in engineered T lymphocytes expressing chimeric antigen receptor (CAR-T), characterizing cancer immunotherapy failure. We conducted this study to investigate whether the combinative application of docetaxel and PSMA-CAR-T cells could be a more effective treatment to prostate cancer. METHODS: Expressions of prostate specific membrane antigen (PSMA) on prostate cancer cells were examined by Flow cytometry. The efficaciousness of PSMA-CAR-T was evaluated in vitro using ELISA and RTCA. The effect of intermixed therapy was assessed in vivo utilizing a human prostate cancer liver metastasis mouse model and a human prostate cancer cell xenograft mouse model. RESULTS: The outcome of cytokine discharge and cell killing assays demonstrated that PSMA-CAR-T cells have characteristic effector capacity against PSMA+ prostate cancer cells in vitro. Additionally, collaborative treatment of PSMA-CAR-T cells and docetaxel have cooperative efficacy in a mouse model of human prostate cancer. The merged strategy could be seen as an undeveloped avenue to augmenting adoptive CAR-T cell immunotherapy and mitigating the adverse side effects of chemotherapy. CONCLUSIONS: Cooperation of PSMA-specific CAR-T cells and the chemotherapy drug docetaxel can impressively ameliorate antitumor effectiveness against an installed metastatic human prostate cancer model in NPG mice.

Essential role of Msx1 in regulating anterior-posterior patterning of the secondary palate in mice.

Development of the secondary palate displays molecular heterogeneity along the anterior-posterior axis; however, the underlying molecular mechanism remains largely unknown. MSX1 is an anteriorly expressed transcription repressor required for palate development. Here, we investigate the role of Msx1 in regional patterning of the secondary palate. The Wnt1-Cre-mediated expression of Msx1 (RosaMsx1Wnt1-Cre) throughout the palatal mesenchyme leads to cleft palate in mice, associated with aberrant cell proliferation and cell death. Osteogenic patterning of the hard palate in RosaMsx1Wnt1-Cre mice is severely impaired, as revealed by a marked reduction in palatine bone formation and decreased expression of the osteogenic regulator Sp7. Overexpression and knockout of Msx1 in mice show that the transcription repressor promotes the expression of the anterior palate-specific Alx1 but represses the expression of the medial-posterior palate genes Barx1, Meox2, and Tbx22. Furthermore, Tbx22 constitutes a direct Msx1 target gene in the secondary palate, suggesting that Msx1 can directly repress the expression of medial-posterior specific genes. Finally, we determine that Sp7 is downstream of Tbx22 in palatal mesenchymal cells, suggesting that a Msx1/Tbx22/Sp7 axis participates in the regulation of palate development. Our findings unveil a novel role for Msx1 in regulating the anterior-posterior growth and patterning of the secondary palate.

RBP and Alb in neonatal hydronephrosis and its association with IL-12 levels in pregnant women before delivery.

Expression levels of retinol-binding protein (RBP) and albumin (Alb) in the kidney of patients with neonatal hydronephrosis were detected to explore whether RBP and Alb can be used as diagnostic indicators for neonatal hydronephrosis. Blood and urine samples from neonates with hydronephrosis in the obstetrics and gynaecology department of Hongqi Hospital Affiliated to Mudanjiang Medical University from January 2016 to January 2018, and healthy newborns were collected. RBP and Alb levels in neonates, and the expression of IL-12 in serum of the mothers were detected by ELISA. To establish the rat model, 30 male Sprague Dawley (SD) rats were randomly divided into two groups: sham operation group (n=15) and unilateral ureteral obstruction (UUO) (n=15). Expression of RBP and Alb protein was detected by western blot analysis. Another 30 female SD rats were randomly divided into the Sham and IL-12 knockdown groups. The Sham group was given NC treatment, while IL-12 knockdown group was given treatment with IL-12 knockdown adenovirus through tail vein injection. Results of ELISA showed that serum RBP and renal urinary Alb levels were higher in neonates with hydronephrosis compared with the control group (P=0.009 or P=0.013). Compared with the control group, levels of IL-12 were significantly lower in mother of neonatal hydronephrosis group (P=0.001). After the IL-12 knockdown treatment in pregnant rats, levels of RBP and Alb hydronephrosis-related indicators were significantly increased. The incidence of hydronephrosis in rats delivered via IL-12 knockdown pregnant rats was significantly higher than that of normal pregnant rats (P=0.001). The combination of RBP and Alb in the diagnosis of neonatal hydronephrosis has important clinical significance, and neonatal hydronephrosis is closely related to the level of IL-12 in mothers before birth. Our findings provide new ideas and theoretical basis for the diagnosis and prediction of neonatal hydronephrosis.

Ectodermal Wnt controls nasal pit morphogenesis through modulation of the BMP/FGF/JNK signaling axis.

BACKGROUND: Mutations of WNT3, WNT5A, WNT9B, and WNT11 genes are associated with orofacial birth defects, including nonsyndromic cleft lip with cleft palate in humans. However, the source of Wnt ligands and their signaling effects on the orofacial morphogenetic process remain elusive. RESULTS: Using Foxg1-Cre to impair Wnt secretion through the inactivation of Gpr177/mWls, we investigate the relevant regulation of Wnt production and signaling in nasal-facial development. Ectodermal ablation of Gpr177 leads to severe facial deformities resulting from dramatically reduced cell proliferation and increased cell death due to a combined loss of WNT, FGF and BMP signaling in the developing facial prominence. In the invaginating nasal pit, the Gpr177 disruption also causes a detrimental effect on migration of the olfactory epithelial cells into the mesenchymal region. The blockage of Wnt secretion apparently impairs the olfactory epithelial cells through modulation of JNK signaling. CONCLUSIONS: Our study thus suggests the head ectoderm, including the facial ectoderm and the neuroectoderm, as the source of canonical as well as noncanonical Wnt ligands during early development of the nasal-facial prominence. Both ß-catenin-dependent and -independent signaling pathways are required for proper development of these morphogenetic processes.

[Factors related to Glycophorin A mutation frequency of workers exposed to benzene.].

OBJECTIVE: To analyze related factors which affect GPA mutation frequency of workers exposed to benzene, with the Glycophorin A (GPA) mutation assay and explore the possibility of GPA mutation frequency as an index of predicting the risk of benzene poisoning. METHODS: The erythrocytes were bound with fluorescent-labeled monoclonal antibody after isolated and fixed from the peripheral blood, and then the GPA mutation assay was performed using the flow cytometry (FCM). The related factors of GPA mutation frequency were analyzed by statistical methods. RESULTS: The GPA mutation frequency of chronic benzene poisonings was significantly higher than that of their controls (P < 0.05). Significant direct correlation was found between age, length of service, accumulative exposure score and the GPA mutation frequency of workers exposed to benzene (P < 0.01). However, there was significantly inverse correlation between the 3AB index and the GPA mutation frequency (GPAN0: r(s) = -0.589, P < 0.01, GPANN: r(s) = -0.615, P < 0.01). In the multiple factor regression analysis on GPA mutation frequency, benzene exposure and individual susceptibility both entered model of multiple factors analysis, the coefficient of determination of benzene-exposed workers was 0.819. CONCLUSION: Exposure to benzene and individual susceptibility are the most important factors that affect GPA mutation frequency. GPA mutation frequency increases with the benzene exposure and individual susceptibility.