Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation.In this study, complementary DNA（cDNA） microarrays were used to identify and study...Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation.In this study, complementary DNA（cDNA） microarrays were used to identify and study the expression of novel genes that may be involved in MC3T3-E1 cells’ response to 17-b estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha（a-MEM）cell culture supplemented with 17-b estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with1028mol？L2117-b estradiol for 5 days exhibited the highest proliferation and alkaline phosphatase（ALP） activity; thus, this group was chosen for microarray analysis. The harvested RNA was used for microarray hybridisation and subsequent real-time reverse transcription polymerase chain reaction（RT-PCR） to validate the expression levels for selected genes. The microarray results were analysed using both functional and pathway analysis. In this study, microarray analysis detected 5 403 differentially expressed genes,of which 1 996 genes were upregulated and 3 407 genes were downregulated, 1 553 different functional classifications were identified by gene ontology（GO） analysis and 53 different pathways were involved based on pathway analysis. Among the differentially expressed genes, a portion not previously reported to be associated with the osteoblast response to oestrogen was identified. These findings clearly demonstrate that the expression of genes related to osteoblast proliferation, cell differentiation, collagens and transforming growth factor beta（TGF-b）-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to a-MEM supplemented with 17-b estradiol. Microarray analysis with functional gene classification is critical for a complete understanding of complementary intracellular processes. This microarray analysis provides large-scale gene expression data that require further confirmatory studies.展开更多
To prove that magnetic characteristics （MC） of participants in breathing are crucial in respiration; to explain the impact of Earth＇s magnetic field （EMF）, cosmic radiation （CR） and MC of cells on evolution and...To prove that magnetic characteristics （MC） of participants in breathing are crucial in respiration; to explain the impact of Earth＇s magnetic field （EMF）, cosmic radiation （CR） and MC of cells on evolution and respiration; to show MC of cells, i.e. development of cytoplasm and membrane of nucleus and cell; to explain crossing over; to show in detail the impact of EMF on cell division; to explain the exchange of oxygen and carbon dioxide in the lungs and cells MC; to explain how the nervous system works as a regulator of respiration. Studying the literature about ceil, respiration and the role of nervous system in breathing; linking natural EMF, CR and MC of cells with evolution and respiration. Every cell has MC. They are very important in every cell metabolism. Crossing over provides polymorphism, enabled by EMF. Crucial factor in cell division is EMF. Electrical pulses （action potentials） are generated by CR. Breathing is enabled by MC of oxygen, carbon dioxide, hemoglobin and cells. MC enables respiration. EMF, CR and MC of cells are crucial factors in evolution processes generally. EMF is the initiator of cell division. Micro electrical pulses from central nervous system are regulators of respiration. Conductivity of micro electrical pulses in the body is fast-electronic and slow-ion （chemical）.展开更多
The reinforcement of calcium phosphate materials with silk fibroin （SF） has been one of the strategies to overcome the brittleness. However, the lack of osteoinductivity may still restrict their further use. This st...The reinforcement of calcium phosphate materials with silk fibroin （SF） has been one of the strategies to overcome the brittleness. However, the lack of osteoinductivity may still restrict their further use. This study aimed to investigate the biocompatibility and osteogenesis capacity of a novel Semaphorin 3A-loaded chitosan microspheres/SF/a-tricalcium phosphate composite （Sema3A CMs/SF/a-TCP） in vitro. Sema3A was first incorporated into CMs, and the Sema3A CMs/SF/a-TCP composite was then prepared. The morphology of the CMs was observed using SEM. The in vitro release kinetics, cytotoxicity, and cell compatibility were evaluated, and the real-time quantitative polymerase chain reaction （RT-qPCR） and activity of alkaline phosphatase （ALP） were used to evaluate the osteogenesis capacity of the composite. The in vitro release of Sema3A from the Sema3A CMs/SF/a-TCP composite showed a temporally controlled manner. The extract of the Sema3A CMs/SF/a-TCP composite presented no obvious side effect on the MC3T3-E1 cell proliferation, nor promote cell proliferation. The MC3T3-E1 cells were well-spread and presented an elongated shape on the Sema3A CMs/SF/a-TCP composite surface; the ALP activity and the osteogenic-related gene expression were higher than those seeded on the surface of the CMs/SF/a-TCP and SF/a-TCP composites. In conclusion, Sema3A CMslSF/a-TCP has excellent biocompatibility and contributes to the osteoblastic differentiation of MC3T3-E1 cells.展开更多
[Objectives]To explore the moxibustion effect and mechanism of"Complementary Acupoints"on rheumatoid arthritis(RA).[Methods]A total of 32 Wistar rats were randomly divided into 4 group:normal control(K),mode...[Objectives]To explore the moxibustion effect and mechanism of"Complementary Acupoints"on rheumatoid arthritis(RA).[Methods]A total of 32 Wistar rats were randomly divided into 4 group:normal control(K),model group(M group),"the Complementary Acupoints"Yanglingquan and Yinlingquan group(AB group),Shenshu and Zusanli group(CD group),with 8 rats in each group.Moxibustion was performed using refined moxa floss(2 mg/pc)on acupoints in both sides every other day,5 pieces for each acupoint,and implemented for 10 times.Before and after the modeling and after the intervention,the left foot sole thermal pain threshold of each group was measured.After the intervention,the rats'left paws were examined by X-ray to detect the number of mast cells(MCs)and degranulation in the affected area of the foot.[Results]The thermal pain threshold of the model group was lower than that of the K group(P<0.05).The number of MC in the affected area and the degranulation were increased.After the moxibustion intervention,the thermal pain threshold of the AB and CD groups was higher than that of the M group(P<0.05);the number of MCs and degranulation were decreased compared with M group,and the degree of reduction was better than that of CD group.The data of foot and claw showed that the foot and paw soft tissue swelling,bone and joint deformity and bone destruction were better in AB and CD groups were more relieved than that in M group.[Conclusions]The moxibustion of Complementary Acupoints can effectively intervene with RA of rats,and its mechanism may be closely related to the MC degranulation.展开更多
Carbon/carbon （C/C） composites were deposited with graphite-like carbon （GLC） coating, and then, Arg-Gly- Asp acid （RGD） peptides were successfully immobilized onto the functionalized GLC coating. GLC coating wa...Carbon/carbon （C/C） composites were deposited with graphite-like carbon （GLC） coating, and then, Arg-Gly- Asp acid （RGD） peptides were successfully immobilized onto the functionalized GLC coating. GLC coating was utilized to prevent carbon particles releasing and create a uniform surface condition for C/C composites. RGD peptides were utilized to improve biocompatibility of GLC coating. Surface chemical characterizations of functionalized GLC coating were detected by contact angle measurement, X-ray photoelectron spectroscopy and Raman spectra. Optical morphology of GLC coatings was observed by confocal laser scanning microscopy. In vitro biological performance was determined using samples seeded with MC3T3-E1 osteoblast-like cells and cultured for 1 week. Surface characterizations and morphological analysis indicated that C/C composites were covered by a dense and uniform GLC coating. Contact angle of GLC coating was reduced to 27.2° when it was functionalized by H202 oxidation at 40 ℃ for 1 h. In vitro cytological test showed that the RGD peptides immobilized GLC coating had a significant improvement in biocompatibility. It was suggested that RGD peptides provided GLC coating with a bioactive surface to improve cell adhesion and proliferation on C/C composites.展开更多
SARS-CoV-2 infection-induced hyper-inflammation is a key pathogenic factor of COVID-19.Our research,along with others',has demonstrated that mast cells(MCs)play a vital role in the initiation of hyper-inflammation...SARS-CoV-2 infection-induced hyper-inflammation is a key pathogenic factor of COVID-19.Our research,along with others',has demonstrated that mast cells(MCs)play a vital role in the initiation of hyper-inflammation caused by SARS-CoV-2.In previous study,we observed that SARS-CoV-2 infection induced the accumulation of MCs in the peri-bronchus and bronchioalveolar-duct junction in humanized mice.Additionally,we found that MC degranulation triggered by the spike protein resulted in inflammation in alveolar epithelial cells and capillary endothelial cells,leading to subsequent lung injury.The trachea and bronchus are the routes for SARS-CoV-2 transmission after virus inhalation,and inflammation in these regions could promote viral spread.MCs are widely distributed throughout the respiratory tract.Thus,in this study,we investigated the role of MCs and their degranulation in the development of inflammation in tracheal-bronchial epithelium.Histological analyses showed the accumulation and degranulation of MCs in the peri-trachea of humanized mice infected with SARS-CoV-2.MC degranulation caused lesions in trachea,and the formation of papillary hyperplasia was observed.Through transcriptome analysis in bronchial epithelial cells,we found that MC degranulation significantly altered multiple cellular signaling,particularly,leading to upregulated immune responses and inflammation.The administration of ebastine or loratadine effectively suppressed the induction of inflammatory factors in bronchial epithelial cells and alleviated tracheal injury in mice.Taken together,our findings confirm the essential role of MC degranulation in SARS-CoV-2-induced hyper-inflammation and the subsequent tissue lesions.Furthermore,our results support the use of ebastine or loratadine to inhibit SARS-CoV-2-triggered degranulation,thereby preventing tissue damage caused by hyper-inflammation.展开更多
Objective: To observe the influence of acupuncture stimulus and thermal stimulus on the expression of mast cells(MCs) and 5-hydroxytryptamine(5-HT) in local acupoint area of Guanyuan(关元 CV 4) and serum 5-HT in rats....Objective: To observe the influence of acupuncture stimulus and thermal stimulus on the expression of mast cells(MCs) and 5-hydroxytryptamine(5-HT) in local acupoint area of Guanyuan(关元 CV 4) and serum 5-HT in rats.Methods: Thirty-nine male C57 BL/6 rats were randomly divided into blank control group, acupuncture stimulus group and thermal stimulus group, with 13 rats in each group. The rats were stimulated by manual acupuncture at CV 4 for 5 min in acupuncture stimulus group, while those in thermal stimulus group were stimulated by adopting a thermal moxibustion apparatus on CV 4 for 30 min. The expression of MC and 5-HT in the skin in the acupoint area of CV 4 before and after acupuncture stimulus and thermal stimulus was observed and analyzed via adopting toluidine blue staining method and immunofluorescence histochemical method(5 rats were selected from each group), and the 5-HT content in serum before and after stimulus was determined through enzyme-linked immunosorbent assay(ELISA)(8 rats were selected from each group). The influence of acupuncture stimulus and thermal stimulus on the contents of MC and 5-HT in the skin and serum 5-HT in rats was analyzed and compared.Results: ① After acupuncture stimulus or thermal stimulus on CV 4 of C57 BL/6 rats, the number of MC in the acupoint area significantly increased when compared with that in blank group(the acupuncture stimulus group 12.40 ± 2.07 vs. the blank group 3.00 ±5.96;thermal stimulus group 26.20 ± 10.85 vs. the blank group 12.40 ± 2.07, both P< 0.05), and MC aggregation and degranulation were observed(the acupuncture stimulus group 17.80 ±4.55 vs. the blank group 8.00 ±3.16;the thermal stimulus group24.00±9.05 vs. the blank group 8.00±3.16. P<0.05, P< 0.01).② After acupuncture stimulus or thermal stimulus, 5-HT was released by MCs in the acupoint area, which aggregated around the blood vessels,and the number of 5-HT in the acupoint area significantly increased when compared with that in blank group(the blank group 3.00 ±1.28 vs. the acupuncture stimulus group 10.02 ±3.21;the blank group3.00 ±1.28 vs. the thermal stimulus group 14.00 ±3.94, both P< 0.01).③ Compared with blank group,both acupuncture stimulus and thermal stimulus could reduce the 5-HT content in serum(the blank group 0.72 ±0.2372 vs. acupuncture stimulus group 0.43 ±0.21: the blank group 0.72 ±0.24 vs. thermal stimulus group 0.32 ±0.18, both P<0.01), and the effect in thermal stimulus group was slightly superior to that in acupuncture stimulus group(P<0.05).Conclusion: Both acupuncture stimulus and thermal stimulus can cause the aggregation and degranulation of MCs and high expression of 5-HT in the acupoint area. The effect of thermal stimulus was superior to that of acupuncture stimulus in degranulation.展开更多
Eucommia ulmoides Oliv.(EuO),also known as Duzhong,native to China,has been reported to have antioxidative function,but its cellular mechanism is not fully examined yet.We investigated inhibitory effects of EuO leaf e...Eucommia ulmoides Oliv.(EuO),also known as Duzhong,native to China,has been reported to have antioxidative function,but its cellular mechanism is not fully examined yet.We investigated inhibitory effects of EuO leaf ethanol extracts on H2O2-induced apoptosis in rat osteoblastic MC3T3-E1 cells and underlying mechanisms.Locally-grown Duzhong leaves were extracted with ethanol.MC3T3-E1 cells were treated with EuO (6.25,12.5,25,50,and 100 μg/ml) for 24 h,and then H2O2 (800 μmol/L) for an additional 24 h.Cell survival rate,percentage of apoptosis,and expressions of caspases 3,6,7,and 9 were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,microscopic analysis,Western blotting,and reverse transcription polymerase chain reaction (RT-PCR).The final EuO leaf ethanol extract powder was detected to contain caffeotannic acid at 58 mg/g and geniposide at 3.45 mg/g by high performance liquid chromatography (HPLC).EuO remarkably restrained cell oxidative damage and increased cell survival rate in a dose-dependent manner: 0 μg/ml,0.21;6.25 μg/ml,0.28;12.5 μg/ml,0.31;25 μg/ml,0.48;50 μg/ml,0.54;and 100 μg/ml,0.66 (P<0.05),with the half-effective concentration being around 25 μg/ml.MTT results were confirmed by microscopic analysis.Western blotting and RT-PCR analyses showed that the expressions of caspases 3,6,7,and 9 were significantly decreased in the EuO-treated cells compared with the control (EuOand H2O2-free) (P<0.05),with the half-effective concentration of EuO ranging from 12.5 to 25 μg/ml.We conclude that the ethanol-extracted EuO leaf extracts promoted the growth of MC3T3-E1 cells,and suppressed the H2O2-induced apoptosis in a rat MC3T3-E1 osteogenic cell model,likely due to the inhibition of caspases’ activities.The results indicate that EuO is a potent antioxidant,which may contribute to its many cellular protective functions,including the promotion of bone growth.展开更多
Alcoholic liver disease(ALD)remains one of the leading causes of liver injury and death when left un-treated.The gut microbiota has been recognized as a key regulator of a number of pathologies,including ALD.The role ...Alcoholic liver disease(ALD)remains one of the leading causes of liver injury and death when left un-treated.The gut microbiota has been recognized as a key regulator of a number of pathologies,including ALD.The role of mast cells(MCs)during liver disease progression has been demonstrated in a number of animal models and in human liver diseases.The interaction between the gut microbiota and MCs has been investigated,and links between the gut and these immune cells are being uncovered.The interplay between the gut microbiota and MCs during ALD has been evaluated and studies suggest that there could be an important link between MCs,their mediators and gut inflammation during the progression of ALD.展开更多
The purpose of this study was to test the hypothesis that the combination of micro-arc oxidation and alkali heatment （MAH） would improve the cytocompatibility of a newly designed Ti-24Nb-4Zr-8Sn alloy. In this study...The purpose of this study was to test the hypothesis that the combination of micro-arc oxidation and alkali heatment （MAH） would improve the cytocompatibility of a newly designed Ti-24Nb-4Zr-8Sn alloy. In this study, commercially pure titanium （cp Ti） and Ti-24Nb-4Zr-8Sn were used. Surface modification of Ti-24Nb- 4Zr-8Sn by a two-step treatment of micro-arc oxidation （MAO） and alkali heatment was reported. Surface characterizations were performed by scanning electron microscopy （SEM）, thin film X-ray diffraction （TF-XRD） and X-ray photoelectron spectroscopy （XPS）. The MAH layer consisted of finer crystals and possessed a higher degree of crystallity and stability than the MAO layer. A biocompatibility study on treated and untreated Ti- 24Nb-4Zr-8Sn in comparison with cp Ti was carried out to investigate the effect of the different surfaces on the bone integration property in vitro. The cellular assays revealed that the MAO and MAH layer favored the initial adhesion of MC3T3-E1 cells and that the growth rate of MC3T3-E1 cells on MAH layer was significantly higher than that on the conventional MAO-treated layer after 3-day and 5-day incubation, demonstrating the greater potential of the hybrid treatment of micro-arc oxidation followed with alkali heatment as a novel surface modification method for implanting materials.展开更多
基金supported by grants from the Natural Science Fund (ZR2010HM035) of Shandong Provincethe Shandong Provincial Health Development Project Fund (2011WSB19002) in China
文摘Oestrogen is essential for maintaining bone mass, and it has been demonstrated to induce osteoblast proliferation and bone formation.In this study, complementary DNA（cDNA） microarrays were used to identify and study the expression of novel genes that may be involved in MC3T3-E1 cells’ response to 17-b estradiol. MC3T3-E1 cells were inoculated in minimum essential media alpha（a-MEM）cell culture supplemented with 17-b estradiol at different concentrations and for different time periods. MC3T3-E1 cells treated with1028mol？L2117-b estradiol for 5 days exhibited the highest proliferation and alkaline phosphatase（ALP） activity; thus, this group was chosen for microarray analysis. The harvested RNA was used for microarray hybridisation and subsequent real-time reverse transcription polymerase chain reaction（RT-PCR） to validate the expression levels for selected genes. The microarray results were analysed using both functional and pathway analysis. In this study, microarray analysis detected 5 403 differentially expressed genes,of which 1 996 genes were upregulated and 3 407 genes were downregulated, 1 553 different functional classifications were identified by gene ontology（GO） analysis and 53 different pathways were involved based on pathway analysis. Among the differentially expressed genes, a portion not previously reported to be associated with the osteoblast response to oestrogen was identified. These findings clearly demonstrate that the expression of genes related to osteoblast proliferation, cell differentiation, collagens and transforming growth factor beta（TGF-b）-related cytokines increases, while the expression of genes related to apoptosis and osteoclast differentiation decreases, following the exposure of MC3T3-E1 cells to a-MEM supplemented with 17-b estradiol. Microarray analysis with functional gene classification is critical for a complete understanding of complementary intracellular processes. This microarray analysis provides large-scale gene expression data that require further confirmatory studies.
文摘To prove that magnetic characteristics （MC） of participants in breathing are crucial in respiration; to explain the impact of Earth＇s magnetic field （EMF）, cosmic radiation （CR） and MC of cells on evolution and respiration; to show MC of cells, i.e. development of cytoplasm and membrane of nucleus and cell; to explain crossing over; to show in detail the impact of EMF on cell division; to explain the exchange of oxygen and carbon dioxide in the lungs and cells MC; to explain how the nervous system works as a regulator of respiration. Studying the literature about ceil, respiration and the role of nervous system in breathing; linking natural EMF, CR and MC of cells with evolution and respiration. Every cell has MC. They are very important in every cell metabolism. Crossing over provides polymorphism, enabled by EMF. Crucial factor in cell division is EMF. Electrical pulses （action potentials） are generated by CR. Breathing is enabled by MC of oxygen, carbon dioxide, hemoglobin and cells. MC enables respiration. EMF, CR and MC of cells are crucial factors in evolution processes generally. EMF is the initiator of cell division. Micro electrical pulses from central nervous system are regulators of respiration. Conductivity of micro electrical pulses in the body is fast-electronic and slow-ion （chemical）.
文摘The reinforcement of calcium phosphate materials with silk fibroin （SF） has been one of the strategies to overcome the brittleness. However, the lack of osteoinductivity may still restrict their further use. This study aimed to investigate the biocompatibility and osteogenesis capacity of a novel Semaphorin 3A-loaded chitosan microspheres/SF/a-tricalcium phosphate composite （Sema3A CMs/SF/a-TCP） in vitro. Sema3A was first incorporated into CMs, and the Sema3A CMs/SF/a-TCP composite was then prepared. The morphology of the CMs was observed using SEM. The in vitro release kinetics, cytotoxicity, and cell compatibility were evaluated, and the real-time quantitative polymerase chain reaction （RT-qPCR） and activity of alkaline phosphatase （ALP） were used to evaluate the osteogenesis capacity of the composite. The in vitro release of Sema3A from the Sema3A CMs/SF/a-TCP composite showed a temporally controlled manner. The extract of the Sema3A CMs/SF/a-TCP composite presented no obvious side effect on the MC3T3-E1 cell proliferation, nor promote cell proliferation. The MC3T3-E1 cells were well-spread and presented an elongated shape on the Sema3A CMs/SF/a-TCP composite surface; the ALP activity and the osteogenic-related gene expression were higher than those seeded on the surface of the CMs/SF/a-TCP and SF/a-TCP composites. In conclusion, Sema3A CMslSF/a-TCP has excellent biocompatibility and contributes to the osteoblastic differentiation of MC3T3-E1 cells.
基金Natural Science Foundation of Hebei Province of China(H2014406047)Scientific and Technological Research Project for Institutions of Higher Education in Hebei Province of China(ZD2019085)Key Scientific Research Project of Chengde Medical University(201812).
文摘[Objectives]To explore the moxibustion effect and mechanism of"Complementary Acupoints"on rheumatoid arthritis(RA).[Methods]A total of 32 Wistar rats were randomly divided into 4 group:normal control(K),model group(M group),"the Complementary Acupoints"Yanglingquan and Yinlingquan group(AB group),Shenshu and Zusanli group(CD group),with 8 rats in each group.Moxibustion was performed using refined moxa floss(2 mg/pc)on acupoints in both sides every other day,5 pieces for each acupoint,and implemented for 10 times.Before and after the modeling and after the intervention,the left foot sole thermal pain threshold of each group was measured.After the intervention,the rats'left paws were examined by X-ray to detect the number of mast cells(MCs)and degranulation in the affected area of the foot.[Results]The thermal pain threshold of the model group was lower than that of the K group(P<0.05).The number of MC in the affected area and the degranulation were increased.After the moxibustion intervention,the thermal pain threshold of the AB and CD groups was higher than that of the M group(P<0.05);the number of MCs and degranulation were decreased compared with M group,and the degree of reduction was better than that of CD group.The data of foot and claw showed that the foot and paw soft tissue swelling,bone and joint deformity and bone destruction were better in AB and CD groups were more relieved than that in M group.[Conclusions]The moxibustion of Complementary Acupoints can effectively intervene with RA of rats,and its mechanism may be closely related to the MC degranulation.
基金supported by the National Natural Science Foundation of China under Grant Nos.51202194 and 51221001the Programme of Introducing Talents of Discipline to Universities(‘‘111’’project of China)under Grant No.B08040
文摘Carbon/carbon （C/C） composites were deposited with graphite-like carbon （GLC） coating, and then, Arg-Gly- Asp acid （RGD） peptides were successfully immobilized onto the functionalized GLC coating. GLC coating was utilized to prevent carbon particles releasing and create a uniform surface condition for C/C composites. RGD peptides were utilized to improve biocompatibility of GLC coating. Surface chemical characterizations of functionalized GLC coating were detected by contact angle measurement, X-ray photoelectron spectroscopy and Raman spectra. Optical morphology of GLC coatings was observed by confocal laser scanning microscopy. In vitro biological performance was determined using samples seeded with MC3T3-E1 osteoblast-like cells and cultured for 1 week. Surface characterizations and morphological analysis indicated that C/C composites were covered by a dense and uniform GLC coating. Contact angle of GLC coating was reduced to 27.2° when it was functionalized by H202 oxidation at 40 ℃ for 1 h. In vitro cytological test showed that the RGD peptides immobilized GLC coating had a significant improvement in biocompatibility. It was suggested that RGD peptides provided GLC coating with a bioactive surface to improve cell adhesion and proliferation on C/C composites.
基金supported by the National Natural Science Foundation of China(82172242)the Natural Science Foundation of Guangdong(2022A1515012053)+2 种基金National Key Research and Development Program of China(2022YFC2303700,2021YFE0113000)Yunnan Key Research and Development Program(202103AC100005)the State key Laboratory of Respiratory Disease,Guangzhou,China(SKLRD-OP202207).
文摘SARS-CoV-2 infection-induced hyper-inflammation is a key pathogenic factor of COVID-19.Our research,along with others',has demonstrated that mast cells(MCs)play a vital role in the initiation of hyper-inflammation caused by SARS-CoV-2.In previous study,we observed that SARS-CoV-2 infection induced the accumulation of MCs in the peri-bronchus and bronchioalveolar-duct junction in humanized mice.Additionally,we found that MC degranulation triggered by the spike protein resulted in inflammation in alveolar epithelial cells and capillary endothelial cells,leading to subsequent lung injury.The trachea and bronchus are the routes for SARS-CoV-2 transmission after virus inhalation,and inflammation in these regions could promote viral spread.MCs are widely distributed throughout the respiratory tract.Thus,in this study,we investigated the role of MCs and their degranulation in the development of inflammation in tracheal-bronchial epithelium.Histological analyses showed the accumulation and degranulation of MCs in the peri-trachea of humanized mice infected with SARS-CoV-2.MC degranulation caused lesions in trachea,and the formation of papillary hyperplasia was observed.Through transcriptome analysis in bronchial epithelial cells,we found that MC degranulation significantly altered multiple cellular signaling,particularly,leading to upregulated immune responses and inflammation.The administration of ebastine or loratadine effectively suppressed the induction of inflammatory factors in bronchial epithelial cells and alleviated tracheal injury in mice.Taken together,our findings confirm the essential role of MC degranulation in SARS-CoV-2-induced hyper-inflammation and the subsequent tissue lesions.Furthermore,our results support the use of ebastine or loratadine to inhibit SARS-CoV-2-triggered degranulation,thereby preventing tissue damage caused by hyper-inflammation.
文摘Objective: To observe the influence of acupuncture stimulus and thermal stimulus on the expression of mast cells(MCs) and 5-hydroxytryptamine(5-HT) in local acupoint area of Guanyuan(关元 CV 4) and serum 5-HT in rats.Methods: Thirty-nine male C57 BL/6 rats were randomly divided into blank control group, acupuncture stimulus group and thermal stimulus group, with 13 rats in each group. The rats were stimulated by manual acupuncture at CV 4 for 5 min in acupuncture stimulus group, while those in thermal stimulus group were stimulated by adopting a thermal moxibustion apparatus on CV 4 for 30 min. The expression of MC and 5-HT in the skin in the acupoint area of CV 4 before and after acupuncture stimulus and thermal stimulus was observed and analyzed via adopting toluidine blue staining method and immunofluorescence histochemical method(5 rats were selected from each group), and the 5-HT content in serum before and after stimulus was determined through enzyme-linked immunosorbent assay(ELISA)(8 rats were selected from each group). The influence of acupuncture stimulus and thermal stimulus on the contents of MC and 5-HT in the skin and serum 5-HT in rats was analyzed and compared.Results: ① After acupuncture stimulus or thermal stimulus on CV 4 of C57 BL/6 rats, the number of MC in the acupoint area significantly increased when compared with that in blank group(the acupuncture stimulus group 12.40 ± 2.07 vs. the blank group 3.00 ±5.96;thermal stimulus group 26.20 ± 10.85 vs. the blank group 12.40 ± 2.07, both P< 0.05), and MC aggregation and degranulation were observed(the acupuncture stimulus group 17.80 ±4.55 vs. the blank group 8.00 ±3.16;the thermal stimulus group24.00±9.05 vs. the blank group 8.00±3.16. P<0.05, P< 0.01).② After acupuncture stimulus or thermal stimulus, 5-HT was released by MCs in the acupoint area, which aggregated around the blood vessels,and the number of 5-HT in the acupoint area significantly increased when compared with that in blank group(the blank group 3.00 ±1.28 vs. the acupuncture stimulus group 10.02 ±3.21;the blank group3.00 ±1.28 vs. the thermal stimulus group 14.00 ±3.94, both P< 0.01).③ Compared with blank group,both acupuncture stimulus and thermal stimulus could reduce the 5-HT content in serum(the blank group 0.72 ±0.2372 vs. acupuncture stimulus group 0.43 ±0.21: the blank group 0.72 ±0.24 vs. thermal stimulus group 0.32 ±0.18, both P<0.01), and the effect in thermal stimulus group was slightly superior to that in acupuncture stimulus group(P<0.05).Conclusion: Both acupuncture stimulus and thermal stimulus can cause the aggregation and degranulation of MCs and high expression of 5-HT in the acupoint area. The effect of thermal stimulus was superior to that of acupuncture stimulus in degranulation.
基金Project (No.2007C33030) supported by the Science and Technology Program of Zhejiang Province,China
文摘Eucommia ulmoides Oliv.(EuO),also known as Duzhong,native to China,has been reported to have antioxidative function,but its cellular mechanism is not fully examined yet.We investigated inhibitory effects of EuO leaf ethanol extracts on H2O2-induced apoptosis in rat osteoblastic MC3T3-E1 cells and underlying mechanisms.Locally-grown Duzhong leaves were extracted with ethanol.MC3T3-E1 cells were treated with EuO (6.25,12.5,25,50,and 100 μg/ml) for 24 h,and then H2O2 (800 μmol/L) for an additional 24 h.Cell survival rate,percentage of apoptosis,and expressions of caspases 3,6,7,and 9 were examined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,microscopic analysis,Western blotting,and reverse transcription polymerase chain reaction (RT-PCR).The final EuO leaf ethanol extract powder was detected to contain caffeotannic acid at 58 mg/g and geniposide at 3.45 mg/g by high performance liquid chromatography (HPLC).EuO remarkably restrained cell oxidative damage and increased cell survival rate in a dose-dependent manner: 0 μg/ml,0.21;6.25 μg/ml,0.28;12.5 μg/ml,0.31;25 μg/ml,0.48;50 μg/ml,0.54;and 100 μg/ml,0.66 (P<0.05),with the half-effective concentration being around 25 μg/ml.MTT results were confirmed by microscopic analysis.Western blotting and RT-PCR analyses showed that the expressions of caspases 3,6,7,and 9 were significantly decreased in the EuO-treated cells compared with the control (EuOand H2O2-free) (P<0.05),with the half-effective concentration of EuO ranging from 12.5 to 25 μg/ml.We conclude that the ethanol-extracted EuO leaf extracts promoted the growth of MC3T3-E1 cells,and suppressed the H2O2-induced apoptosis in a rat MC3T3-E1 osteogenic cell model,likely due to the inhibition of caspases’ activities.The results indicate that EuO is a potent antioxidant,which may contribute to its many cellular protective functions,including the promotion of bone growth.
基金Portions of this work were supported by(i)a VA Merit Award(1I01BX003031,HF)from the United States Department of Veter-an's AffairsBiomedical Laboratory Research and Development Service and(ii)the R01 grant from NIH NIDDK(DK108959,HF).
文摘Alcoholic liver disease(ALD)remains one of the leading causes of liver injury and death when left un-treated.The gut microbiota has been recognized as a key regulator of a number of pathologies,including ALD.The role of mast cells(MCs)during liver disease progression has been demonstrated in a number of animal models and in human liver diseases.The interaction between the gut microbiota and MCs has been investigated,and links between the gut and these immune cells are being uncovered.The interplay between the gut microbiota and MCs during ALD has been evaluated and studies suggest that there could be an important link between MCs,their mediators and gut inflammation during the progression of ALD.
基金supported by the National Natural Science Foundation of China under grant No. 30973354
文摘The purpose of this study was to test the hypothesis that the combination of micro-arc oxidation and alkali heatment （MAH） would improve the cytocompatibility of a newly designed Ti-24Nb-4Zr-8Sn alloy. In this study, commercially pure titanium （cp Ti） and Ti-24Nb-4Zr-8Sn were used. Surface modification of Ti-24Nb- 4Zr-8Sn by a two-step treatment of micro-arc oxidation （MAO） and alkali heatment was reported. Surface characterizations were performed by scanning electron microscopy （SEM）, thin film X-ray diffraction （TF-XRD） and X-ray photoelectron spectroscopy （XPS）. The MAH layer consisted of finer crystals and possessed a higher degree of crystallity and stability than the MAO layer. A biocompatibility study on treated and untreated Ti- 24Nb-4Zr-8Sn in comparison with cp Ti was carried out to investigate the effect of the different surfaces on the bone integration property in vitro. The cellular assays revealed that the MAO and MAH layer favored the initial adhesion of MC3T3-E1 cells and that the growth rate of MC3T3-E1 cells on MAH layer was significantly higher than that on the conventional MAO-treated layer after 3-day and 5-day incubation, demonstrating the greater potential of the hybrid treatment of micro-arc oxidation followed with alkali heatment as a novel surface modification method for implanting materials.
