1.Logistics regression analysis of plastic bronchitis in children with mycoplasma pneumoniae infection
Fen OU ; Taoyi YANG ; Guanglei CHEN ; Hongxia LI ; Pingping LI
Journal of Public Health and Preventive Medicine 2025;36(1):160-163
Objective To analyze the influencing factors of plastic bronchitis in children with Mycoplasma pneumoniae infection and put forward targeted prevention suggestions. Methods The clinical data of children with Mycoplasma pneumoniae infection who were admitted to Chengdu Third People's Hospital from September 2022 to February 2024 were retrospectively analyzed . According to whether plastic bronchitis occurred, they were divided into plastic group (n=118) and non-plastic group (n=184), and the differences between the two groups were compared and analyzed. Univariate and multivariate logistics regression analysis equations were used to analyze the independent influencing factors of plastic bronchitis in children with mycoplasma pneumoniae infection. Results Among the 302 children with Mycoplasma pneumoniae infection , 118 cases were diagnosed with plastic bronchitis. Analysis showed that the children’s age, duration of fever, hospital stay, pleural effusion rate, number of bronchoscopic lavage, allergy history, endoscopic mucosal erosion rate, WBC, NE%, LY%, CRP, LDH, PCT and D-D were the single factors influencing the occurrence of plastic bronchitis in children with mycoplasma pneumoniae infection. Binary logistics regression analysis revealed that age (OR=2.137, P=0.033, 95% CI: 1.132-16.603), allergy history (OR=3.028, P=0.014, 95% CI: 1.261-864), NE% (OR=2.395, P=0.031, 95% CI: 1.087-5.274), CRP (OR=3.864, P=0.004, 95% CI: 1.563-3.864), PCT (OR=4.125, P=0.001, 95% CI: 1.793-3.864), and D-D (OR=3.920, P=0.002, 95% CI: 1.632-3.864) were independent risk factors for plastic bronchitis in children with mycoplasma pneumoniae infection (P<0.05). Conclusion Age, allergy history, NE%, CRP, PCT and D-D are independent risk factors for plastic bronchitis in children with mycoplasma pneumoniae infection . It is necessary to take clinical intervention measures to reduce the occurrence risk.
2.Inner Ear Delivery of Polyamino Acid Nanohydrogels Loaded with Dexamethasone
Pingping AI ; Lidong ZHAO ; Zhaohui TANG ; Chaoliang HE ; Xuesi CHEN ; Shiming YANG ; Nan WU
Medical Journal of Peking Union Medical College Hospital 2025;16(2):370-378
To develop a novel polyamino acid-based nanohydrogel drug delivery system for dexamethasone to enhance its delivery efficiency to the inner ear. A fluorescein-labeled polyglutamic acid-based polyamino acid dexamethasone nanohydrogel was synthesized, and its gelation time was measured. The hydrogel was surgically injected into the round window niche of guinea pigs to determine its degradation time in the middle ear cavity in vivo. The safety, pharmacokinetics, and distribution patterns of dexamethasone in the inner ear were evaluated. The hydrogel exhibited a gelation time of 80 seconds in a 37℃ water bath. In vivo, the hydrogel was almost completely degraded within 7 days in the middle ear cavity of guinea pigs. Transient hearing loss was observed one day after administration, but hearing gradually returned to normal over time. No significant cytotoxicity, vestibular stimulation signs, or pathological abnormalities in spiral ganglion cells were observed. Histopathological examination revealed no significant inflammatory reactions. Pharmacokinetic analysis demonstrated sustained drug release and prolonged dexamethasone activity. Immunofluorescence staining confirmed the distribution of dexamethasone in both the cochlea and vestibular organs. The polyamino acid nanohydrogel exhibits excellent injectability and biodegradability, representing a safe and effective drug delivery system for the inner ear.
3.Treatment of Recurrent Aphthous Ulcers from the Perspective of "Heart"
Mengfan REN ; Nailin ZHANG ; Ruohan WANG ; Mengqian SUN ; Pingping CHEN ; Hua CAO ; Qiquan LIU
Journal of Traditional Chinese Medicine 2025;66(11):1174-1177
Based on the traditional Chinese medicine theory that "all pain, itching, and sores are related to the heart", this paper proposes treating recurrent aphthous ulcers from the perspective of the heart. It suggests that excessive heart fire and tissue erosion due to flaming fire in the heart meridian constitute the core pathogenesis of this condition. Hyperactive heart fire is identified as the key pathogenic factor, while heart yin deficiency, obstruction of the heart collaterals, and malnourishment of the heart spirit are considered significant contributing factors. Clinically, the treatment follows the principle of clearing heart fire as the main strategy, supplemented by nourishing yin, activating collaterals, and calming the spirit. The self-formulated Qingxin Yuchuang Formulation (清心愈疮方) serves as the base prescription, with flexible modifications incorporating the Yuyin Formulation (育阴方), Huoxue Formulation (活血方), and Yu'an Formulation (郁安方) to address specific syndromes involving heart yin deficiency, collateral blockage, and emotional disturbance.
4.Effect of Dictamni Cortex on Intestinal Barrier Damage by Untargeted Metabolomics and Targeted Metabolomics for Short-chain Fatty Acids
Xiaomin XU ; Donghua YU ; Yu WANG ; Pingping CHEN ; Jiameixue WO ; Suxia JIA ; Wenkai HU ; Fang LU ; Shumin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(20):40-47
ObjectiveThis study aims to investigate the effect of Dictamni Cortex on intestinal barrier damage in rats and its mechanism by untargeted metabolomics and targeted metabolomics for short-chain fatty acids (SCFAs). MethodsRats were randomly divided into a control group, a high-dose group of Dictamni Cortex (8.1 g·kg-1), a medium-dose group (2.7 g·kg-1), and a low-dose group (0.9 g·kg-1). Except for the control group, the other groups were administered different doses of Dictamni Cortex by gavage for eight consecutive weeks. Hematoxylin-eosin (HE) staining was used to observe the pathological changes in the ileal tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to detect the level of cytokines, including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β), in the ileal tissue of rats. Quantitative real-time fluorescence polymerase chain reaction (Real-time PCR) technology was used to detect the expression level of tight junction proteins, including zonula occludens-1 (ZO-1), Occludin, and Claudin-1 mRNAs, in the ileal tissue of rats to preliminarily explore the effects of Dictamni Cortex on intestinal damage. The dose with the most significant toxic phenotype was selected to further reveal the effects of Dictamni Cortex on the metabolic profile of ileal tissue in rats by non-targeted metabolomics combined with targeted metabolomics for SCFAs. ResultsCompared with the control group, all doses of Dictamni Cortex induced varying degrees of pathological damage in the ileum, increased TNF-α (P<0.01), IL-6 (P<0.01), and IL-1β (P<0.01) levels in the ileal tissue, and decreased the expression level of ZO-1 (P<0.05, P<0.01), Occludin (P<0.01), and Claudin-1 (P<0.05) in the ileal tissue, with the high-dose group showing the most significant toxic phenotypes. The damage mechanisms of the high-dose group of Dictamni Cortex on the ileal tissue were further explored by integrating non-targeted metabolomics and targeted metabolomics for SCFAs. The non-targeted metabolomics results showed that 21 differential metabolites were identified in the control group and the high-dose group. Compared with that in the control group, after Dictamni Cortex intervention, the level of 14 metabolites was significantly increased (P<0.05, P<0.01), and the level of seven metabolites was significantly decreased (P<0.05, P<0.01) in the ileal contents. These metabolites collectively acted on 10 related metabolic pathways, including glycerophospholipids and primary bile acid biosynthesis. The quantitative data of targeted metabolomics for SCFAs showed that Dictamni Cortex intervention disrupted the level of propionic acid, butyric acid, acetic acid, caproic acid, isobutyric acid, isovaleric acid, valeric acid, and isocaproic acid in the ileal contents of rats. Compared with those in the control group, the level of isobutyric acid, isovaleric acid, and valeric acid were significantly increased, while the level of propionic acid, butyric acid, and acetic acid were significantly decreased in the ileal contents of rats after Dictamni Cortex intervention (P<0.05, P<0.01). ConclusionDictamni Cortex can induce intestinal damage by regulating glycerophospholipid metabolism, primary bile acid biosynthesis, and metabolic pathways for SCFAs.
5.Underlying Mechanism of Wuwei Shenqintang in Amelioration of Pulmonary Fibrosis by Regulating "Lung-intestine Axis" Based on UPLC-Q-TOF-MS Metabolomics Technology
Mengdi SUN ; Fang LU ; Donghua YU ; Yu WANG ; Pingping CHEN ; Shumin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):11-20
ObjectiveTo explore the mechanism of action of Wuwei Shenqintang in improving pulmonary fibrosis by using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) for metabolomic analysis of lung tissue and feces. MethodsA rat model with pulmonary fibrosis was established by intratracheal injection of 5 mg·kg-1 bleomycin. The successfully modeled rats were randomly divided into a blank group, a model group, a prednisone (3.15 mg·kg-1) group, and low-dose, medium-dose, and high-dose groups of Wuwei Shenqintang (4.586, 9.172, 18.344 g·kg-1). The rats were given intragastric administration once a day for 28 consecutive days. Hematoxylin-eosin (HE) staining was used to measure the pathological changes in lung and colon tissue, and Masson staining was used to detect the degree of pulmonary fibrosis. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of interleukin-1β (IL-1β), IL-6, IL-8, tumor necrosis factor-α (TNF-α), and secretory immunoglobulin A (SIgA) in bronchoalveolar lavage fluid and intestinal mucus. Immunohistochemistry and reverse transcription quantitative polymerase chain reaction (Real-time PCR) were used to detect the expression of type Ⅰ collagen (Col-Ⅰ), fibronectin (FN), and alpha smooth muscle actin (α-SMA) in lung tissue. UPLC-Q-TOF-MS was used to study the changes in the metabolic network of lung tissue and feces in rats with pulmonary fibrosis treated with Wuwei Shenqintang, screen potential biomarkers for the treatment of pulmonary fibrosis by Wuwei Shenqintang, and perform pathway enrichment analysis. ResultsCompared with the blank group, the model group showed extensive inflammatory cell infiltration and continuous fibrotic lesions in lung tissue, colonic mucosal damage, and connective tissue hyperplasia. The expression of IL-6, IL-8, IL-1β, TNF-α, and SIgA in bronchoalveolar lavage fluid and intestinal mucus was significantly increased (P<0.01). The expression of Col-Ⅰ, FN, and α-SMA proteins and mRNAs in lung tissue was significantly upregulated (P<0.01). Compared with the model group, the groups of Wuwei Shenqintang exhibited significantly reduced inflammatory infiltration and blue collagen deposition in lung tissue, alleviated colonic damage, decreased expression of IL-6, IL-8, IL-1β, TNF-α, and SIgA in bronchoalveolar lavage fluid and intestinal mucus (P<0.01), and reduced average absorbance values and mRNA expression of Col-Ⅰ, FN, and α-SMA in lung tissue (P<0.05, P<0.01), with the prednisone group and the medium-dose and high-dose groups of Wuwei Shenqintang showing the most significant effects. The metabolomics results for lung tissue showed that compared with the blank group, the model group had 19 significantly different compounds (P<0.05, P<0.01). Wuwei Shenqintang could normalize 17 of these compounds compared with the model group (P<0.05, P<0.01). Fecal metabolomics results showed that compared with those in the blank group, there were 42 compounds with significant differences in the model group (P<0.05, P<0.01). Compared with the model control group, Wuwei Shenqintang could normalize 41 of these compounds (P<0.05, P<0.01). The combined analysis results indicated that Wuwei Shenqintang might inhibit pulmonary fibrosis by regulating the biosynthesis of phenylalanine, tyrosine, and tryptophan as well as the retinol metabolism pathway. ConclusionWuwei Shenqintang can ameliorate pulmonary fibrosis, which may be related to the regulation of the "lung-intestine axis".
6.Underlying Mechanism of Wuwei Shenqintang in Amelioration of Pulmonary Fibrosis by Regulating "Lung-intestine Axis" Based on UPLC-Q-TOF-MS Metabolomics Technology
Mengdi SUN ; Fang LU ; Donghua YU ; Yu WANG ; Pingping CHEN ; Shumin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):11-20
ObjectiveTo explore the mechanism of action of Wuwei Shenqintang in improving pulmonary fibrosis by using ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) for metabolomic analysis of lung tissue and feces. MethodsA rat model with pulmonary fibrosis was established by intratracheal injection of 5 mg·kg-1 bleomycin. The successfully modeled rats were randomly divided into a blank group, a model group, a prednisone (3.15 mg·kg-1) group, and low-dose, medium-dose, and high-dose groups of Wuwei Shenqintang (4.586, 9.172, 18.344 g·kg-1). The rats were given intragastric administration once a day for 28 consecutive days. Hematoxylin-eosin (HE) staining was used to measure the pathological changes in lung and colon tissue, and Masson staining was used to detect the degree of pulmonary fibrosis. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of interleukin-1β (IL-1β), IL-6, IL-8, tumor necrosis factor-α (TNF-α), and secretory immunoglobulin A (SIgA) in bronchoalveolar lavage fluid and intestinal mucus. Immunohistochemistry and reverse transcription quantitative polymerase chain reaction (Real-time PCR) were used to detect the expression of type Ⅰ collagen (Col-Ⅰ), fibronectin (FN), and alpha smooth muscle actin (α-SMA) in lung tissue. UPLC-Q-TOF-MS was used to study the changes in the metabolic network of lung tissue and feces in rats with pulmonary fibrosis treated with Wuwei Shenqintang, screen potential biomarkers for the treatment of pulmonary fibrosis by Wuwei Shenqintang, and perform pathway enrichment analysis. ResultsCompared with the blank group, the model group showed extensive inflammatory cell infiltration and continuous fibrotic lesions in lung tissue, colonic mucosal damage, and connective tissue hyperplasia. The expression of IL-6, IL-8, IL-1β, TNF-α, and SIgA in bronchoalveolar lavage fluid and intestinal mucus was significantly increased (P<0.01). The expression of Col-Ⅰ, FN, and α-SMA proteins and mRNAs in lung tissue was significantly upregulated (P<0.01). Compared with the model group, the groups of Wuwei Shenqintang exhibited significantly reduced inflammatory infiltration and blue collagen deposition in lung tissue, alleviated colonic damage, decreased expression of IL-6, IL-8, IL-1β, TNF-α, and SIgA in bronchoalveolar lavage fluid and intestinal mucus (P<0.01), and reduced average absorbance values and mRNA expression of Col-Ⅰ, FN, and α-SMA in lung tissue (P<0.05, P<0.01), with the prednisone group and the medium-dose and high-dose groups of Wuwei Shenqintang showing the most significant effects. The metabolomics results for lung tissue showed that compared with the blank group, the model group had 19 significantly different compounds (P<0.05, P<0.01). Wuwei Shenqintang could normalize 17 of these compounds compared with the model group (P<0.05, P<0.01). Fecal metabolomics results showed that compared with those in the blank group, there were 42 compounds with significant differences in the model group (P<0.05, P<0.01). Compared with the model control group, Wuwei Shenqintang could normalize 41 of these compounds (P<0.05, P<0.01). The combined analysis results indicated that Wuwei Shenqintang might inhibit pulmonary fibrosis by regulating the biosynthesis of phenylalanine, tyrosine, and tryptophan as well as the retinol metabolism pathway. ConclusionWuwei Shenqintang can ameliorate pulmonary fibrosis, which may be related to the regulation of the "lung-intestine axis".
7.Single-center retrospective analysis of efficacy and safety of daratumumab plus dexamethasone for light chain amyloidosis nephropathy
Han XUE ; Yaomin WANG ; Liangliang CHEN ; Quan HAN ; Pingping REN ; Lan LAN ; Guangjun LIU ; Jianghua CHEN ; Fei HAN
Chinese Journal of Nephrology 2024;40(1):4-10
Objective:To analyze the efficacy and safety of daratumumab plus dexamethasone in the treatment of renal injury patients with light chain amyloidosis, and to provide clinical reference.Methods:It was a single center retrospective observational study. The clinical data before and after daratumumab treatment of renal injury patients with light chain amyloidosis treated with daratumumab plus dexamethasone from December 2021 to August 2022 were retrospectively collected. The hematologic response, kidney response, prognosis, and adverse events were analyzed. The treatment regimen was 16 mg/kg intravenous infusion of daratumumab on day 1 + 20 mg intravenous push of dexamethasone on day 1-2, once every 2 weeks. The follow-up was up to February 28, 2023.Results:The study included 18 patients, with age of (58.4±7.7) years old, and a male to female ratio of 11∶7. Eleven patients were newly diagnosed and 7 patients were retreated. There were 7, 5, 5 and 1 patients, respectively at the stage Ⅰ, Ⅱ, Ⅲ and Ⅳ of light chain amyloidosis according to 2012 Mayo stage criteria. The median course of disease before onset was 2.5 (1.0, 8.0) months and the follow-up time was (8.7±2.8) months. The patients received (10±3) times of treatment. The overall hematologic response rates were 9/13, 11/13 and 13/13 at 1 month, 3 months, and 6 months respectively after treatment, meanwhile 8/13, 10/13 and 12/13 achieved at least very good partial response at 1 month, 3 months, and 6 months respectively (the other 5 patients did not undergo detailed evaluation due to baseline difference of serum free κ and λ light chain <20 mg/L). The median duration of hematologic response was 16 (13, 40) days. At 3 months, 6 months and the end of follow-up, 10, 13 and 13 of 18 patients respectively achieved renal response, and the median duration of response was 66 (26, 182) days. During follow-up, the median difference of serum free κ and λ light chain decreased by 93% (72%, 97%). Until the last follow-up, one patient died of organ hemorrhage. Other infusion reactions, leukopenia, neutropenia and infection all improved after symptomatic treatments.Conclusion:Daratumumab plus dexamethasone treatment is effective for light chain amyloidosis nephropathy in inducing hematologic remission and kidney remission, with good safety.
8.Distribution and seasonal fluctuation of visceral leishmaniasis vectors sandflie in Lüliang City of Shanxi Province in 2023
Yanqin WANG ; Yujie JIA ; Yifang LIANG ; Pingping HAO ; Xiaodong TIAN ; Wei FENG ; Xiaoqing DONG ; Caiping WANG ; Yanbin ZHANG ; Huize CHEN
Chinese Journal of Schistosomiasis Control 2024;36(4):403-406
Objective To investigate the distribution and seasonal fluctuations of visceral leishmaniasis vectors sandflies in Lüliang City, Shanxi Province, so as to provide insights into assessment of the visceral leishmaniasis transmission risk and formulation of visceral leishmaniasis control measures. Methods A total of 12 natural villages were sampled from Shilou County, Lishi District, Lanxian County, Linxian County and Wenshui County in Lüliang City, Shanxi Province from June to September, 2023, and sandflies were captured using light traps from 7 breeding habitats, including farmers’ houses, sheep pens, cattle pens, chicken coops, pig pens, mule and horse pens, and loess-cave dwellings. Following morphological identification of the sandfly species, the distribution of sandflies and the seasonal fluctuations of the sandfly density were analyzed. In addition, the Leishmania was detected in sandflies using a real-time fluorescence quantitative PCR assay. Results A total of 2 831 sandflies were captured with 156 light traps in Lüliang City from June to September, 2023, including 2 638 female sandflies (93.18%) and 193 male sandflies (6.82%), and the average density was 16.91 sandflies/(light-night). The seasonal fluctuations of the sandfly density all appeared a unimodal distribution in all survey sites, and the sandfly density peaked in July and then declined rapidly. Among all types of breeding habitats, the greatest sandfly density was found in sheep pens [39.04 sandflies/(light-night)]. In addition, 4.08% (2/49) of the sandfly samples were tested positive for Leishmania nucleic acid as revealed by the real-time fluorescence quantitative PCR assay. Conclusions Sandflies were widely distributed in Lüliang City, Shanxi Province in 2023, and the peak of the sandfly density was observed in July, which had a visceral leishmaniasis transmission risk. Intensified surveillance of visceral leishmaniasis and sandfly vectors is required and targeted vector control is recommended.
9.Effect of Cannabidiol on Pulmonary Fibrosis Analyzed by UPLC-Q-TOF-MS
Mengdi SUN ; Feiyu ZHANG ; Huicong YANG ; Yu WANG ; Pingping CHEN ; Fang LU ; Shumin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(12):185-193
ObjectiveTo investigate the mechanism of anti-pulmonary fibrosis of cannabidiol by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). MethodSD rats were randomly divided into blank group, model group, prednisone group(3.15 mg·kg-1) and cannabidiol low, medium and high dose groups(12, 36, 108 mg·kg-1), with 8 rats in each group. The rat model of pulmonary fibrosis was established by intratracheal injection of bleomycin(5 mg·kg-1), which was administered continuously for 28 days after successful modeling. The pathological changes of rat lung tissue were observed, and enzyme-linked immunosorbent assay(ELISA) was used to detect the expression levels of matrix metalloproteinase-7(MMP-7), type Ⅱ alveolar cell surface antigen(KL-6), pulmonary surfactant-associated protein A(SP-A) and SP-D in serum. The expression levels of type Ⅰ collagen(Col-Ⅰ) and fibronectin(FN) in lung tissues were detected by immunohistochemistry, and the expression of mucin 5 subtype AC(MUC5AC) was detected by immunofluorescence. UPLC-Q-TOF-MS was used to search for potential biomarkers and related metabolic pathways of cannabidiol in treating pulmonary fibrosis. ResultCompared with the blank group, there were a large number of inflammatory cell infiltration and continuous fibrosis lesions in the lung tissue of rats in the model group. Compared with the model group, the inflammatory infiltration and blue collagen deposition in the lung tissue of rats in the prednisone and cannabidiol groups were reduced. Compared with the blank group, the expressions of MMP-7, KL-6, SP-A and SP-D in serum of the model group were significantly increased(P<0.01), while the expressions of MMP-7, KL-6, SP-A and SP-D in the prednisone and cannabidiol high dose groups were significantly decreased by comparing with the model group(P<0.05, P<0.01). Compared with the blank group, the expression levels of Col-Ⅰ and FN in the lung tissues of the model group were significantly increased, and the fluorescence intensity of MUC5AC was significantly increased(P<0.01). Compared with the model group, the expression levels of Col-Ⅰ and FN in the lung tissues of the prednisone and cannabidiol high dose groups were significantly decreased(P<0.05, P<0.01), and the expression of MUC5AC was significantly decreased(P<0.01). Compared with the blank group, a total of 18 differential compounds were screened out in the model group, which could be used as potential biomarkers, and cannabidiol could call back 16 of them, mainly involving 4 metabolic pathways(linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, and niacin and niacinamide metabolism). Compared with the blank group, the relative contents of potential biomarkers arachidonic acid and linoleic acid were significantly increased in the model group(P<0.05, P<0.01), while the relative contents of 5,6-EET, L-tyrosine and niacinamide were significantly decreased(P<0.01). Compared with the model group, cannabidiol could significantly reduce the relative contents of arachidonic acid and linoleic acid, and significantly increase the relative contents of 5,6-EET, L-tyrosine and niacinamide(P<0.01). ConclusionCannabidiol has an intervention and remission effect on pulmonary fibrosis, and its mechanism may be related to linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, niacin and niacinamide metabolism.
10.Effects of spheroid culture in a rotating bioreactor on secretion of inflammatory factors by human placenta-derived mesenchymal stem cells
Pingping ZHANG ; Tingting LIANG ; Mingsong FAN ; Li CHEN ; Shichang ZHANG
Chinese Journal of Tissue Engineering Research 2024;28(19):3012-3017
BACKGROUND:Spheroid culture of mesenchymal stem cells in bioreactors is an in vitro culture method to maintain their stemness properties and allow for large-scale expansion.Clarifying its effects on the immunoregulation effect of stem cells is beneficial for their clinical application. OBJECTIVE:To investigate the effects of spheroid culture in a rotating bioreactor on the secretion of inflammatory factors by human placenta-derived mesenchymal stem cells. METHODS:Placenta-derived mesenchymal stem cells isolated from human placenta tissue were cultured in two-dimensional culture or a rotating bioreactor culture.Cell morphology and proliferation ability were observed using inverted phase contrast microscopy,immunohistochemical staining,and CCK-8 assay.The gene expression and protein secretion of several inflammatory factors were detected by RT-qPCR and flow immunofluorescence assay. RESULTS AND CONCLUSION:(1)Mesenchymal stem cells cultured in a rotating bioreactor aggregated into multicellular spheroids,which gradually increased in number and volume.(2)The hematoxylin-eosin staining results showed that the mesenchymal stem cells cultured in the rotating bioreactor for 4 days were evenly distributed and had normal morphology in the spheroids.(3)Immunohistochemical staining results revealed many mesenchymal stem cells with Ki-67 positive in the spheroids.(4)The CCK-8 assay results exhibited that the viability of mesenchymal stem cells derived from spheroid culture was significantly higher than that of cells cultured in two-dimensional culture.(5)The results of RT-qPCR and flow immunofluorescence assay demonstrated that the gene expression and protein secretion(interleukin 1β,interleukin-4,interleukin-6,interleukin-8,interleukin-10,interleukin-17,tumor necrosis factor α and interferon α)of inflammatory factors derived from mesenchymal stem cells cultured in the rotating bioreactor were significantly higher than those in two-dimensional culture.(6)Our results indicate that spheroid culture in a rotating bioreactor can significantly elevate the secretion ability of various inflammatory factors by human placenta-derived mesenchymal stem cells,and enhance the immunoregulatory effect of human placenta-derived mesenchymal stem cells.


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