Objective:To investigate the effect of microwave on human adenovirus type 2 (HAdV-2) capsid protein in simulated infectious wastes.Methods:Droplets of HAdV-2 virus suspension were added to medical disposable gloves to simulate infectious waste, irradiated under different microwave conditions, the temperature change was recorded, and the irradiated viral supernatant was treated with Dnase I and detected by PCR and qPCR to determine the effect of microwave on the integrity of the viral capsid. ELISA was used to detect the effect of microwave irradiation on the structure of viral hexon protein. The virus was treated alone at the highest temperature during microwave irradiation to investigate whether there were non-thermal effects during microwave disinfection.Results:The maximum temperature during microwave irradiation was 76 ℃, and the PCR and qPCR result showed that Dnase I could significantly damage the viral nucleic acid after microwave irradiation, while the virus control group and heat treatment group were not significantly affected, indicating that microwave irradiation could destroy the integrity of the viral capsid. The result of ELISA showed that microwave irradiation could significantly weaken the binding ability of Hexon protein and antibody, and the result of heat treatment group were similar.Conclusions:Microwave irradiation can destroy the integrity of the HAdV-2 capsid and the structure of Hexon protein, in which the damage to the integrity of the capsid is mainly due to non-thermal effects, and the structural changes of hexon protein are mainly due to thermal effects.

Aim To explore the mechanism of oxidized lipoprotein(a)(oxLp(a))inducing pyroptosis of vascu-lar endothelial cells.Methods After incubating human umbilical vein endothelial cells(HUVEC)with 100 mg/L ox-Lp(a)for 24 hours,Western blot and RT-qPCR was used to detect pyroptosis related proteins,pro-inflammatory cytokines,mitochondrial related proteins NRF1,NRF2,PGC-1α and mitochondrial gene cytochrome b(CYTB),ELISA was used to detect the levels of inflammatory factors,scanning electron microscopy was used to detect cell membrane rup-ture,transmission electron microscopy was used to detect mitochondrial morphology,Hoechst33342/PI staining was used to detect cell apoptosis,MitoSOX probe was used to detect mitochondrial reactive oxygen species(mtROS),Flu-4AM probe was used to detect calcium ions,JC-1 probe was used to detect mitochondrial membrane potential(MMP),and Calcein AM staining was used to detect mitochondrial permeability transition pore(mPTP).Transfecting HUVEC with CYTB overexpressing lentivirus and analyzing its effects on oxLp(a)induced pyroptosis and mitochondrial function.Results After treatment with oxLp(a),the expression of NLRP3,pro-Caspase-1,Caspase-1,GSDMD and GSDMD-N proteins re-lated to pyroptosis were significantly increased(P＜0.05);the protein and mRNA levels of CYTB and pro-inflammatory cy-tokine IL-1β,IL-18 were significantly increased(P＜0.05).Small pores appeared on the cell membrane,the percentage of PI stained positive cells significantly increased(P＜0.05).OxLp(a)significantly inhibited the expression of mito-chondrial related proteins NRF1,NRF2 and PGC-1α,and the expression of mitochondrial gene CYTB,promoted an in-crease in mtROS generation,Ca2+overload,a decrease in ATP levels,a decrease in MMP,an increase in mPTP values,and abnormal mitochondrial morphology.After transfection with pHelper 2.0 lentivirus vector overexpressing CYTB,it was found that oxLp(a)induced HUVEC pyroptosis and mitochondrial morphological and functional abnormalities were par-tially reversed by overexpression of CYTB.Conclusion oxLp(a)promotes mitochondrial morphological and functional abnormalities and induces HUVEC pyroptosis by downregulating CYTB.

Objective To investigate the repairing effect and mechanism of Chinese patent medicine Weiyangning pill on gastric mucosal injury in rats induced by anhydrous ethanol,and to establish a high-performance liquid chromatography(HPLC)method to determine the five main components of Weiyangning pill.Methods The five components of paeoniflorin,psoralen,atractylenolide Ⅲ,liquiritin and hesperidin in Weiyangning pill were detected by HPLC.SD male rats were randomly divided into normal control group,model control group,Weinaian group,and large and small dose group of Weiyangning pill.All rats were fasted for 24 hours without water fasting.The normal control group and the model control group were given purified water by gavage.While Weinaian group was given Weinaian(3 g·kg-1),the test group were given intragastric perfusion of Weiyangning(3,1.5 g·kg-1)respectively.After 2 hours,all the rats,except the normal control group,were intragastrically administered with anhydrous ethanol(5 mL·kg-1)to establish the model of gastric mucosal injury.An hour later,the experimental materials were collected,and the gross score of gastric mucosal injury was observed and calculated.The gastric mucosal slices were stained by hematoxylin-eosin(HE)to calculate the pathological scores.Immunohistochemistry was employed to detect the expression of gastric mucosa-related proteins.Results The high-performance liquid chromatogram of Weiyangning pill was obtained,and the absorption peaks with the same retention time as the five standard substances(paeoniflorin,psoralen,atractylenolide Ⅲ,liquiritin and hesperidin)were observed.The general score and pathological score of gastric mucosal injury in Weiyangning groups(3,1.5 g·kg-1)were lower than those of the model control group(P<0.05 or P<0.01).Weiyangning pill(3,1.5 g·kg-1)ameliorated the decrease expression of tight junction protein(Claudin-7),adhesion junction proteins(E-cadherin and β-catenin),mucins(MUC1 and MUC5AC),and the gastric transcription factor SOX2 in the gastric mucosa of the rats modeled in anhydrous ethanol(P<0.05 or P<0.01 compared with the model control group).Conclusion The repairing effect of Weiyangning pill on gastric mucosal injury induced by anhydrous ethanol in rats is related to the increase of the expression of tight junction protein,adhesion junction protein,mucin and gastric transcription factor.

OBJECTIVE To evaluate the impact of sublingual immunotherapy(SLIT)on the balance of Treg/Th17 cells and related cytokines in preschool children aged 3-6 years with allergic rhinitis(AR).METHODS Seventy preschool children aged 3-6 years with AR were divided into the SLIT group and the medication group,and their clinical data were retrospectively analyzed.The medication group received symptomatic treatment alone,while the SLIT group received a combined treatment of SLIT and symptomatic medication,with a 3-year follow-up period.The Treg/Th17 cell balance,serum levels of TGF-β,IL-10,IL-17,IL-21,as well as the total nasal symptom score(TNSS),total medication score(TMS),and visual analog scale(VAS)scores were compared before and after treatment between the two groups.RESULTS After 3 years of treatment,both groups showed significant improvements(P<0.05)in the percentages of CD4+CD25+Foxp3+Treg and CD4+IL-17+Th17 cells among CD4+T cells,percentages of Treg and Th17 cells,serum levels of TGF-β,IL-10,IL-17,IL-21,TNSS,TMS,and VAS scores.Moreover,the SLIT group exhibited significantly better outcomes compared to the medication group(P<0.05).CONCLUSION SLIT can modulate the balance of Treg/Th17 cells and improve serum levels of TGF-β,IL-10,IL-17,and IL-21.

Objective:To study the inactivation effects of microwave on human adenovirus 2 (HAdV-2) in simulated infectied wastes, and to explore its molecular mechanism.Methods:25 μl of HAdV-2 virus suspension was dripped with medical disposable gloves, masks and cotton swabs to simulate infectied wastes, and irradiated under different microwave conditions: gloves and masks were irradiated for 30 s, 60 s, and 90 s at 300 W, 500 W, and 700 W, respectively. Cotton swabs irradiate 60 s, 90 s, 120 s at 500 W and 700 W respectively. Temperature changes were recorded, and the inactivation logarithmic values were calculated by the 50% endpoint method to evaluate the microwave inactivation effects, and the proliferation ability of the virus was detected by qPCR. The damage of Penton, Fiber, Hexon and E2 B genes was detected by PCR. The virus was treated with the highest temperature of 76 ℃ during microwave irradiation to study whether there was non-thermal effect during microwave disinfection. Results:After microwave irradiation of infectied waste, the temperature of masks and gloves carriers rises rapidly, with the highest temperature of 76 ℃. The temperature of the cotton swab carriers rose slowly, and the highest temperature is 65 ℃. The inactivation effect of microwave on HAdV-2 was positively correlated with microwave power and irradiation time. In the mask and glove group, microwave power of 700 W irradiated for 60 seconds, and the inactivation logarithm value could reach 3.0, In the cotton swab group, microwave power of 700 W irradiated for 120 seconds, and the inactivation logarithm value was still less than 3.0. This indicated that there were differences in the conditions for microwave inactivation of the virus in different carriers. The qPCR result showed that microwave irradiation could weak the proliferation ability of the virus. Microwave irradiation had no effect on the virus's Penton and Fiber genes, but caused some damage to the Hexon and E2 B genes. The inactivation effect of individual heat treatment on HAdV-2 was weaker than that of microwave irradiation, and there was no damage to the Hexon, Penton, Fiber, and E2 B genes. This indicated the presence of non thermal effects during the microwave inactivation process. Conclusions:Microwave irradiation can inactivate HAdV-2 in simulated infectied wastes through thermal and non-thermal effects, and its damage to viral DNA is one of the mechanisms of virus inactivation.

Although the mTOR-4E-BP1 signaling pathway is implicated in aging and aging-related disorders, the role of 4E-BP1 in regulating human stem cell homeostasis remains largely unknown. Here, we report that the expression of 4E-BP1 decreases along with the senescence of human mesenchymal stem cells (hMSCs). Genetic inactivation of 4E-BP1 in hMSCs compromises mitochondrial respiration, increases mitochondrial reactive oxygen species (ROS) production, and accelerates cellular senescence. Mechanistically, the absence of 4E-BP1 destabilizes proteins in mitochondrial respiration complexes, especially several key subunits of complex III including UQCRC2. Ectopic expression of 4E-BP1 attenuates mitochondrial abnormalities and alleviates cellular senescence in 4E-BP1-deficient hMSCs as well as in physiologically aged hMSCs. These f indings together demonstrate that 4E-BP1 functions as a geroprotector to mitigate human stem cell senescence and maintain mitochondrial homeostasis, particularly for the mitochondrial respiration complex III, thus providing a new potential target to counteract human stem cell senescence.
Mesenchymal Stem Cells/physiology* ; Cellular Senescence ; Homeostasis ; Cell Cycle Proteins/metabolism* ; Adaptor Proteins, Signal Transducing/metabolism* ; Mitochondria/metabolism* ; Electron Transport Complex III/metabolism* ; Humans ; Cells, Cultured

Humans ; RNA, Long Noncoding/genetics* ; MicroRNAs/genetics* ; Cell Line, Tumor ; Inflammation/genetics* ; Apolipoproteins E ; Apoptosis

Objective    To explore the safety and effectiveness of different interventional approaches for the treatment of patent ductus arteriosus (PDA) in children. Methods    The children (≤7 years) who underwent interventional treatment for PDA from 2019 to 2020 in our hospital were retrospectively included. The patients were divided into 3 groups according to the procedures: a conventional arteriovenous approach group, a simple venous approach group, and a retrograde femoral artery approach group. The clinical efficacy of the patients was compared. Results     A total of 220 patients were included. There were 78 males and 142 females, with an average age of 3.21±1.73 years, weight of 14.99±5.35 kg, and height of 96.19±15.77 cm. The average diameter of the PDA was 3.35±1.34 mm. A total of 85 patients received a conventional arteriovenous approach, 104 patients received a simple venous approach, and 31 patients received a retrograde femoral artery approach. The diameter of PDA in the retrograde femoral artery group was smaller than that in the other two groups (3.44±1.43 mm vs. 1.99±0.55 mm; 3.69±1.17 mm vs. 1.99±0.55 mm, P<0.001); the contrast medium usage [40 (30, 50) mL vs. 20 (20, 30) mL; 35 (25, 50) mL vs. 20 (20, 30) mL, P≤0.001] and operation time [32 (26, 44) min vs. 25 (23, 30) min; 29 (25, 38) min vs. 25 (23, 30) min, P<0.05] in the simple venous approach group were significantly less or shorter than those in the other two groups; the length of hospital stay of the conventional arteriovenous group was longer than that in the other two groups [3 (3, 5) d vs. 4 (3, 6) d; 4 (3, 5) d vs. 4 (3, 6) d, P<0.05]. There was no significant difference in postoperative complications. Conclusion    It is safe and effective to close PDA through simple venous approach. The retrograde femoral artery approach has the advantage of simplifying the surgical procedure for PDA with small diameters.

Cell Differentiation ; Humans ; Hyperthermia, Induced ; Stem Cells

Humans ; Mechanistic Target of Rapamycin Complex 2/metabolism* ; Multiprotein Complexes/metabolism* ; Neural Stem Cells/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Rapamycin-Insensitive Companion of mTOR Protein/metabolism* ; TOR Serine-Threonine Kinases/metabolism*