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ObjectiveTo reveal the clinical efficacy of Yunpi Huatan Tongqiao prescription in relieving inflammation， hypoxia， and adenoidal hypertrophy （AH）， and improving the quality of sleep-disordered breathing in children with AH by promoting M2-type polarization of macrophages through a randomized double-blind clinical trial. MethodSeventy-one AH children who met the research criteria and were treated in the Department of Pediatrics of Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from March 2022 to February 2023 were divided into an observation group （36 cases） and a control group （35 cases）. A randomized double-blind method was adopted. The patients in the observation group were given Yunpi Huatan Tongqiao prescription combined with placebo of montelukast sodium chewable tablets， while those in the control group were given montelukast sodium chewable tablets combined with placebo of Yunpi Huatan Tongqiao prescription. The treatment course was 84 days. After treatment， the therapeutic effect and sleep monitoring indicators were compared. Before and after treatment， venous blood was collected to compare the levels of macrophage polarization-related inflammatory factors between the two groups. ResultThe adenoidal/nasopharyngeal space （A/N） integral in the nasal and pharyngeal lateral radiographs， After treatment， the AH therapeutic effect score， and the traditional Chinese medicine （TCM） syndrome therapeutic effect score in both groups were lower than those before treatment （P<0.01）. Compared with the control group after treatment， the observation group showed a more significant reduction in various integral levels （P<0.05， P<0.01）. The improvement degree of A/N in the nasal and pharyngeal lateral radiographs in the observation group was better than that in the control group （Z=-2.970， P<0.01）， and the total effective rate of the therapeutic effect of AH （χ2=7.715， P<0.01） and the TCM syndrome therapeutic effect （χ2=13.239， P<0.01） were superior to those in the control group. The levels of interleukin-6 （IL-6） and tumor necrosis factor-alpha （TNF-α） in both groups after treatment were lower， and the level of interleukin-10 （IL-10） was higher than those before treatment （P<0.01）. The level of transforming growth factor-beta （TGF-β） showed an increasing trend， but the difference was not statistically significant. Compared with the control group after treatment， the observation group showed more significant improvement in IL-10 and TNF-α levels （P<0.01）， a decreasing trend in IL-6， and an increasing trend in TGF-β， but the difference was not statistically significant. Compared with the results before treatment， the apnea-hypopnea index （AHI） and oxygen desaturation index （ODI） in both groups decreased significantly （P<0.01）. The observation group showed a significant reduction in the duration of the longest apnea and the longest hypopnea， as well as a significant increase in the mean and lowest oxygen saturation （P<0.01）. The control group also showed improvements in the above indicators， but the difference was not statistically significant. Compared with the control group after treatment， the observation group showed a more significant improvement in AHI， ODI， the duration of the longest hypopnea， and mean and lowest oxygen saturation （P<0.05， P<0.01）. There was a decreasing trend in the longest duration of apnea， but the difference was not statistically significant. ConclusionYunpi Huatan Tongqiao prescription can reduce the size of adenoids， alleviate clinical symptoms and signs in AH children， improve the constitution characterized by "spleen deficiency and phlegm obstruction"， reduce the occurrence of sleep-disordered breathing events， alleviate the degree of hypoxia in the child's body during sleep at night， and has satisfactory clinical efficacy. The improvement of clinical symptoms and sleep quality in AH children by Yunpi Huatan Tongqiao prescription may be achieved by promoting macrophage polarization from M1 to M2.
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{L-End}Objective To estimate the number of workers with occupational noise-induced hearing loss (ONHL) in the manufacturing industry of China in 2020. {L-End}Methods Multiple data sources were integrated to collect information on workers in the manufacturing industry from the Occupational Diseases and Hazards Monitoring Information System under China Disease Prevention and Control Information System. The total number of workers was split based on age from the 2018 Fourth National Economic Census and the 2020 National Survey of Occupational Disease Hazards. Additionally, data on the prevalence of hearing loss in the general population was adjusted based on the age composition of Jilin Province, and noise prevalence was standardized based on the age composition of the employed population in the Seventh National Population Censu. Attribution fractions (AF) was estimated. The prevalence ratio (PR) was calculated by the prevalence of hearing loss in the occupational noise-exposed workers and general population. The proportion and attributable cases (AC) of ONHL cases in all hearing loss cases were estimated for occupational noise-exposed workers. The number of ONHL was estimated based on AF and the total number of workers with hearing loss. {L-End}Results In 2020, a total of 30 059 525 workers were exposed to occupational noise in the manufacturing industry in China, with noise-exposed prevalence and standardized noise-exposed prevalence of 28.94% and 28.52%, respectively. The prevalence of hearing loss among occupational noise-exposed workers increased with age, and a total of 8 054 789 workers suffered from hearing loss. Most of the cases were at the age between 45-<55 years old. The total PR and 95% uncertainty interval (UI) was 2.83 (2.58-3.06) and the total AF and 95%UI was 64.63% (61.22%-67.30%), and both peaks were in the age of 30-<45 years. The AC and 95%UI was 5 205 980 (4 930 620-5 420 345) persons, and most of the cases were at the age between 40-<55 years. {L-End}Conclusion Occupational noise poses a serious threat to the hearing health of workers in the manufacturing industry of China, especially among middle-aged and young adults.
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Objective To evaluate the effectiveness of reactive oxygen species (ROS)-responsive nanomedicine in suppressing corneal neovascularization (CNV) in vivo.Methods ROS-responsive nanomedicine (ROS-TK-5/siVEGF),which consists of vascular endothelial growth factor (VEGF) small interfering RNA (siRNA) and thioketal linkage was synthesized by the Michael addition.The cumulative release of siVEGF from nanomedicine under oxidant conditions was assessed by agarose gel electrophoresis.Thirty-nine VEGFR2-1uc-KI transgenic mice were used in this study,of which 30 mice were randomly divided into a normal control group,a PBS control group,an ROS-TK-5/NC group,an ROS-TK-5/siVEGF group,and a ranibizumab group,with 6 mice in each group.The ROS levels in the corneal tissue after alkali burning were tested by dihydroethidium (DHE) staining in the other 9 mice.In each group,alkali-burned mice were subconjunctivally injected with 10 μl of a different formula every two days.The effectiveness of nanomedicine in attenuating CNV was evaluated by slit-lamp microscopy and an in vivo imaging system (IVIS) at 7,14,and 21 days after alkali burning.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology (ARVO) and the Guidelines of the Animal Experimental Committee of Liberation Army General Hospital.The study protocol was approved by the Ethics Committee of Liberation Army General Hospital (No.2018-X14-82).Results After treathrent with an aqueous solution without ROS,only 5％-10％ of the siVEGF was released from the nanoparticles within 10 hours.In contrast,about 70％ of the siVEGF was released from the nanoparticles after treatment with 10 mmol/L H2O2 within 10 hours.The relative fluorescent intensities in the corneal stromal layer at 7 days and 14 days after alkali burning were 5.403±0.306 and 2.930±0.255,respectively,which was significantly greater than those in the normal control group (1.003±0.015) (both at P＜0.05).The CNV areas were statistically different among the four groups at various time points (Fgroup =49.855,P＜0.01;Ftime =65.556,P＜0.01).The CNV area was significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared with the PBS control and ROS-TK-5/NC groups at 7 days and 14 days after modeling,and the CNV area was more effectively reduced in the ROS-TK-5/siVEGF group than the ranibizumab group at 7 days and 14 days after modeling (all at P＜0.05).At day 21 after modeling,the CNV area was significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups (all at P＜ 0.05).IVIS showed that the corneal fluorescent intensity was statistically different among the four groups at various times (Fgroup =27.193,P =0.003;Ftime =51.062,P ＜ 0.01).The corneal fluorescent intensities were significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups at 7 days and 14 days after modeling;in addition,the corneal fluorescent intensity was more effectively reduced in the ROS-TK-5/siVEGF group in comparison with the ranibizumab group at 7 days and 14 days after modeling (all at P＜ 0.05).At 21 days after modeling,the corneal fluorescent intensity was significantly reduced in the ROS-TK-5/ siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups (all at P ＜ 0.05).Conclusions ROS-TK-5/siVEGF nanomedicine effectively attenuates alkali burn-induced CNV formation and appears to have a better effect in comparison with ranibizumab at an early stage.
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Macrophages play an important role in material-related immune responses and bone formation, but the functionality of macrophage-derived extracellular vesicles (EVs) in material-mediated bone regeneration is still unclear. Here, we evaluated intracellular communication through small extracellular vesicles (sEVs) and its effects on endogenous bone regeneration mediated by biomimetic intrafibrillarly mineralized collagen (IMC). After implantation in the bone defect area, IMC generated more neobone and recruited more mesenchymal stem cells (MSCs) than did extrafibrillarly mineralized collagen (EMC). More CD63
Subject(s)
Biomimetics , Bone Regeneration , Cell Differentiation , Collagen , Extracellular Vesicles , Macrophages , OsteogenesisABSTRACT
Objective@#To evaluate the effectiveness of reactive oxygen species (ROS)-responsive nanomedicine in suppressing corneal neovascularization (CNV) in vivo.@*Methods@#ROS-responsive nanomedicine (ROS-TK-5/siVEGF), which consists of vascular endothelial growth factor (VEGF) small interfering RNA (siRNA) and thioketal linkage was synthesized by the Michael addition.The cumulative release of siVEGF from nanomedicine under oxidant conditions was assessed by agarose gel electrophoresis.Thirty-nine VEGFR2-luc-KI transgenic mice were used in this study, of which 30 mice were randomly divided into a normal control group, a PBS control group, an ROS-TK-5/NC group, an ROS-TK-5/siVEGF group, and a ranibizumab group, with 6 mice in each group.The ROS levels in the corneal tissue after alkali burning were tested by dihydroethidium (DHE) staining in the other 9 mice.In each group, alkali-burned mice were subconjunctivally injected with 10 μl of a different formula every two days.The effectiveness of nanomedicine in attenuating CNV was evaluated by slit-lamp microscopy and an in vivo imaging system (IVIS) at 7, 14, and 21 days after alkali burning. The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology (ARVO) and the Guidelines of the Animal Experimental Committee of Liberation Army General Hospital.The study protocol was approved by the Ethics Committee of Liberation Army General Hospital (No.2018-X14-82).@*Results@#After treathrent with an aqueous solution without ROS, only 5%-10% of the siVEGF was released from the nanoparticles within 10 hours.In contrast, about 70% of the siVEGF was released from the nanoparticles after treatment with 10 mmol/L H2O2 within 10 hours.The relative fluorescent intensities in the corneal stromal layer at 7 days and 14 days after alkali burning were 5.403±0.306 and 2.930±0.255, respectively, which was significantly greater than those in the normal control group (1.003±0.015) (both at P<0.05). The CNV areas were statistically different among the four groups at various time points (Fgroup=49.855, P<0.01; Ftime=65.556, P<0.01). The CNV area was significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared with the PBS control and ROS-TK-5/NC groups at 7 days and 14 days after modeling, and the CNV area was more effectively reduced in the ROS-TK-5/siVEGF group than the ranibizumab group at 7 days and 14 days after modeling (all at P<0.05). At day 21 after modeling, the CNV area was significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups (all at P<0.05). IVIS showed that the corneal fluorescent intensity was statistically different among the four groups at various times (Fgroup=27.193, P=0.003; Ftime=51.062, P<0.01). The corneal fluorescent intensities were significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups at 7 days and 14 days after modeling; in addition, the corneal fluorescent intensity was more effectively reduced in the ROS-TK-5/siVEGF group in comparison with the ranibizumab group at 7 days and 14 days after modeling (all at P<0.05). At 21 days after modeling, the corneal fluorescent intensity was significantly reduced in the ROS-TK-5/siVEGF and ranibizumab groups compared to the PBS control and ROS-TK-5/NC groups (all at P<0.05).@*Conclusions@#ROS-TK-5/siVEGF nanomedicine effectively attenuates alkali burn-induced CNV formation and appears to have a better effect in comparison with ranibizumab at an early stage.
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Esophagus, as the pipe connecting oral cavity and stomach, has unique anatomical structure and physiological functions. The related diseases including esophageal cancer and precancerous lesions are the ones of major public health problems in China. The pathogenesis of esophageal diseases is still not clear. The exploration of correlation between the changes of esophageal microbiota and esophageal diseases becomes a new breakthrough in the study of etiology. Previous studies have found enrichment of gram-positive bacteria in the normal esophagus, while a decrease in diversity of bacteria and a dominant gram-negative anaerobe in diseased esophagus. Although much progress has been made in the study of esophageal microbiota, the standard method of how to accurately and noninvasively collect esophageal microbiota is still lack, which is an important part of the esophageal microbial research.
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OBJECTIVE@#To investigate the effects of millimeter wave (MMW) exposure on apoptosis of human melanoma A375 cells and explore the mechanisms.@*METHODS@#Through electromagnetic field calculation we simulated MMW exposure in cells and calculated the specific absorption rate (SAR). The optimal irradiation parameters were determined according to the uniformity and intensity of the SAR. A375 cells were then exposed to MMV for 15, 30, 60, or 90 min, with or without pretreatment with the caspase-3 inhibitor AC-DEVD-fmk (10 μmol/L) for 1 h at 90 min before the exposure. CCK-8 assay was used to assess the changes in the viability and Annexin-V/ PI staining was used to detect the apoptosis of the cells following the exposures; Western blotting was used to detect the expression of caspase-3 in the cells.@*RESULTS@#The results of electromagnetic field calculation showed that for optimal MMV exposure, the incident field needed to be perpendicular to the bottom of the plastic Petri dish with the antenna placed below the dish. CCk-8 assay showed that MMW exposure significantly inhibited the cell viability in a time-dependent manner ( < 0.05); exposures for 15, 30, 60, and 90 min all resulted in significantly increased apoptosis of the cells ( < 0.05). The cells with MMW exposure showed significantly increased expression of caspase-3. The inhibitory effect of MMW on the cell viability was antagonized significantly by pretreatment of the cells with AC-DEVD-fmk ( < 0.05), which increased the cell viability rate from (36.7±0.09)% to (59.8±0.06)% ( < 0.05).@*CONCLUSIONS@#35.2 GHz millimeter wave irradiation induces apoptosis in A375 cells by activating the caspase-3 protein.
Subject(s)
Humans , Apoptosis , Caspase 3 , Metabolism , Caspase Inhibitors , Pharmacology , Cell Line, Tumor , Cell Survival , Electromagnetic Fields , Enzyme Activation , Magnetic Field Therapy , Melanoma , Pathology , Therapeutics , Time FactorsABSTRACT
Objective@#To investigate the effect of cell-to-cell communication amongst single-cell clones from healthy periodontium with different osteogenic differentiation potentials on change of osteogenic differentiation capabilities and the role histone acetyltransferase partaken in this process.@*Methods@#In order to research the change of osteogenic differentiation ability via cell-to-cell communication, indirect co-culture method was used by placing two single-cell clones with different osteogenesis potentials in each of the 6-well plates. Blank control, weak and strong osteogenic groups were set up, corresponding to Transwell chambers with blank, cells of weak osteogenesis ability and cells of strong osteogenesis ability, respectively. Each group was made in triplicate. After co-culture for four days, Transwell chamber was removed. Quantitative real-time PCR (qPCR) and alizarin red staining were employed to detect the change of osteogenic differentiation ability. The acetylation level of H3 was measured by using Western blotting. Histone acetyltransferases were detected by qPCR.@*Results@#Single-cell clones were ensured from mesenchymal stem cells by flow cytometer, the positive expression of CD29, CD90, CD105, CD146 was (99.80±0.02)%, (99.36±0.18)%, (99.41±0.05)% and (95.10±2.11)%, respectively. And CD31 and CD34 expression were (0.29±0.11)% and (0.22±0.13)%, respectively. Alizarin red and oil red O staining confirmed that single-cell clones had the abilities of adipogenesis and osteogenesis. Alkaline phosphatase (ALP) and alizarin red staining indicated that different single-cell clones were heterogeneity in osteogenesis differentiation. Indirect co-culture indicated that the mRNA expression of osteocalcin (OCN) were 14.24±5.60 and 4.78±2.90, respectively and Runt-related transcription factor 2 (RUNX2) were 2.75±1.44 and 1.61±0.44, respectively, in strong and weak osteogenic groups. They were significantly higher compared to the blank group (the mRNA expression of OCN and RUNX2 were 1.00±0.47 and 1.00±0.39, respectively). The expression of OCN and RUNX2 were also higher in strong osteogenic group than that in weak osteogenic group (P<0.05). The mean gray level of the acetylation of H3 in strong osteogenic group (0.76±0.09) and weak osteogenic group (0.54±0.12) were also higher than that in the blank group (0.30±0.04)(P<0.05). qPCR results showed that KAT6A in strong osteogenic group exhibiting higher expression (P<0.05) compared to weak osteogenic group and the blank group, which were corresponding to the changes of acetylation levels.@*Conclusions@#Single-cell clones from healthy periodontium showed heterogeneity in osteogenic differentiation abilities. Single-cell clones with strong osteogenesis abilities had an advantage over others by promoting others' osteogenesis differentiation and this change mediated by cell-to-cell communication might be caused by modulating KAT6A to affect the acetylation level of histone.