1.The comparison on pluripotent differentiation between human gingival fibroblasts and human periodontal ligament cells in vitro
Rui SU ; Liting SONG ; Yunyun DONG ; Jiayin DENG ; Shaoyun JIANG
Tianjin Medical Journal 2016;44(2):137-141
Objective To investigate the the multi-directional differentiation potential between pluripotent of human gingival fibroblasts (HGFs) and human periodontal ligament cells (HPDLCs). Methods HPDLCs and HGFs were obtained from the primary culture. HPDLCs and HGFs at 3rd-4th passage were cultured in osteogenic, adipogenic or chondrogenic me-dium. Cells without differentiation were taken as control. Alizarin red, Alcian blue and oil red O staining were performed to detect osteogenic differentiation, chondrogenic and adipogenic differentiation in vitro, respectively. Reverse transcription polymerase chain reaction (RT-PCR) was applied to examine the expression of osteocalcin (OCN), runt-related transcription factor 2 (RUNX2) and collagen 1 (Col 1), peroxisome proliferator-activated receptor gamma 2 (PPARγ2) and collagen 10 (Col 10). Results HPDLCs and HGFs cultured in osteogenic medium showed massive calicium nodulus at day 28, but HP-DLCs formed more calicium nodulus than those of HGFs. The expressions of OCN, RUNX2 and Col 1 were significantly high-er in HPDLCs than those in HGFs (P<0.05). In chondrogenic medium both cells were found blue deposit at day 14, and the expression of Col 10 was significantly higher in HGFs than that of HPDLCs (P<0.01). Furthermore, in adipogenic medium HGFs showed more lipid-filled droplets stained with oil red O than HPDLCs at day 21. The expression of PPARγ2 was sig-nificantly higher in HGFs than that of HPDLCs (P<0.01). Conclusion HPDLCs has the better potency of osteogenic differ-etiation than HGFs, however, HGFs has the better potency of adipogenic and chondrogenic differentiation.
2.Determination of Perfluorinated Compounds in Water by Electrospray Ionization Mass Spectrometry Combined with Hollow Fiber Membrane Extraction
Yao YAO ; Jiewei DENG ; Yunyun YANG ; Ling FANG ; Haiyun ZHOU ; Hongtao LIU ; Xiaowei WANG ; Tiangang LUAN
Chinese Journal of Analytical Chemistry 2015;(7):1053-1057
In this study, hollow fiber membrane extraction combined with ambient ionization mass spectrometry ( AMS) was developed for the simultaneous determination of 7 perfluorinated compounds ( PFCs) in aqueous solution, including perfluoroheptanoic acid ( PFHpA ) , perfluorooctanoic acid ( PFOA ) , perfluorooctane sulfonate acid ( PFOS ) , perfluorononanoic acid ( PFNA ) , perfluorodecanoic acid ( PFDA ) , perfluoroundecanoic acid ( PFUdA) , and perfluorododecanoic acid ( PFDoA) . PFCs were detected in negative ion mode using selective reaction monitoring ( SRM) mode. The extraction time and the pH value of extraction solution were optimized. 13 C4-PFOS and 13 C4-PFOA were used as internal standards for quantitative analysis. The method showed good linearity with correlation coefficient values ( r2 ) greater than 0. 991 for the seven target PFCs. With the exception of PFHpA, the limit of detection ( LOD) for other six PFCs was within ranges from 0. 8 to 2. 7 ng/L while the limit of quantitative (LOQ) was from 2. 7 ng/L to 8. 9 ng/L. The enrichment factor of five PFCs was more than two hundred. The developed method was applied to detect the seven PFCs in tap water and Pearl River water, and they were all not detected. The recoveries were within the ranges of 88. 5%-108. 3% and 94. 2%-116. 7% when 40 ng/L and 400 ng/L PFCs were spiked into tap water, respectively. In terms of the Pearl River water, the recoveries were within the ranges of 75. 0%-102. 6% and 81. 2%-97. 6% when 40 ng/L and 400 ng/L PFCs were spiked, respectively.
3.Effect of high glucose on the secretion of cytokines induced by Porphyromonas gingivalis lipopolysaccharide.
Shaoyun JIANG ; Congcong WEI ; Dong XUE ; Jiayin DENG ; Qi LIAN ; Yunyun DONG
Chinese Journal of Stomatology 2014;49(12):737-741
OBJECTIVETo investigate the influence of high glucose on Porphyromonasgingivalis (Pg) lipopolysaccharide (LPS) stimulating human gingival fibroblasts (HGF) to secret the cytokines.
METHODSHGF were obtained from the primary culture of the tissue explants. Cells were divided into four groups, low glucose (5.5 mmol/L) + 1 mg/L Pg LPS (group A);low glucose + 10 mg/L Pg LPS (group B); high glucose (25 mmol/L) +1 mg/L Pg LPS(group C); high glucose+10 mg/L Pg LPS (group D). The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in cell supernatants were detected by enzyme- linked immunosorbent assay at 6 h and 12 h. The expressions of toll-like receptor 2, 4 (TLR-2, 4) were examined by real-time polymerase chain reaction. After pretreatment with anti-TLR2 and anti-TLR4 monoclonal antibody in HGF, TNF-α and L-1β levels were detected.
RESULTSTNF-α concentration increased obviously in high glucose 6 h and 12 h after Pg LPS stimulation (P < 0.01). IL-1β secretion increased (P < 0.01). Meanwhile, TLR2, 4 mRNA expression increased, especially in high glucose+10 mg/L Pg LPS (P < 0.01). After inhibition of the TLR2, 4 in high glucose + 10 mg/L Pg LPS respectively, TNF-α level [(297.16±11.49), (390.01±12.81) ng/L] decreased (F = 166.02, P < 0.01), and IL-1β level [(49.90±4.08), (99.35±5.01) ng/L] also decreased (F = 153.51, P < 0.01).
CONCLUSIONSHigh glucose may promote Pg LPS to stimulate the secretion of TNF-α and IL-1β through regulating TLR2, 4 expression, which suggests that the elevating blood glucose precipitate in aggravating the process of periodontal disease.
Antibodies, Monoclonal ; Drug Synergism ; Fibroblasts ; drug effects ; metabolism ; Glucose ; administration & dosage ; pharmacology ; Humans ; Interleukin-1beta ; metabolism ; Periodontal Diseases ; etiology ; Polysaccharides, Bacterial ; toxicity ; Porphyromonas gingivalis ; chemistry ; Time Factors ; Toll-Like Receptor 2 ; immunology ; metabolism ; Toll-Like Receptor 4 ; immunology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism
4.Spatiotemporal distribution characteristics of polychlorinated dibenzo-p-dioxins and dibenzofurans in PM2.5 in Pudong of Shanghai and population health assessment
Wanlian MAO ; Lijuan JIA ; Xiaoyu SHEN ; Xiaoqian CHEN ; Yuxin ZHU ; Pinchuan YANG ; Yunyun DENG
Journal of Environmental and Occupational Medicine 2024;41(5):482-488
Background In the Global Burden of Disease research, it has been found that atmospheric fine particulate matter (PM2.5) pollution significantly harms human health. Currently, there is limited research on polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) that exhibit high toxicity effects in PM2.5 . Objective By studying the spatiotemporal distribution and variation characteristics of PCDD/Fs in PM2.5 in Pudong area of Shanghai, to assess the associated population health risk. Methods This study set up 28 sampling points in Pudong area. One sample of PM2.5 was collected during winter (February 2022) and summer (July 2022) at each site, with a sampling period lasting 24 h. The concentration of PM2.5 was measured by membrane filter method, and the content of 17 kinds of 2,3,7,8-substituted chlorinated PCDD/Fs in the samples was analyzed using isotope dilution. Seasonal variations (winter and summer) in the concentrations of PM2.5 and PCDD/Fs were evaluated, sources of PCDD/Fs pollution were tracing by principal component analysis, and health risks to the population from respiratory exposure to PCDD/Fs were estimated by VLIER-HUMAAN model. Results The PM2.5 concentrations in the 28 samples ranged from 10 to 126 μg·m−3, while the concentrations of PCDD/Fs in PM2.5 ranged from 58 to 2625 fg·m−3. The concentration of PM2.5 during winter (11-126 μg·m−3) was higher than that during summer (10-60 μg·m−3). The concentration range of PCDD/Fs in winter was from 58 to 2625 fg·m−3, which corresponded to a range of toxic equivalent quantity (WHO-TEQ) concentration from 2.99 to 40.97 fg·m−3 when taking World Health Organization's toxic equivalency factor (WHO-TEQ); the concentration range of PCDD/Fs in summer was from 72 to 446 fg·m−3, which corresponded to a range of WHO-TEQ concentration from 2.66 to 16.61 fg·m−3. This range in summer was significantly lower than that observed in winter. The results of principal component analysis revealed that waste incineration was the primary source of PCDD/Fs in winter PM2.5 in the area, whereas traffic emissions emerged as the main source in summer. The assessment of Pudong residents' respiratory exposure to PCDD/Fs in PM2.5 showed significantly higher exposure of children in summer and winter than that of adults, indicating higher susceptibility of children to air pollutants. Both the hazard ratios (HR) for children and adults were below 1, while the cancer risks (CR) ranged from 8.41×10−8 to 2.35×10−6. Notably, during winter, the CR at 4 locations slightly exceeds 1×10−6, indicating a potential carcinogenic risk. Conclusion The overall pollution level of PCDD/Fs in PM2.5 in Pudong area is relatively low, but it shows clear seasonal patterns. Waste incineration and traffic are the main sources of PCDD/Fs in PM2.5 in the area. Although the cancer risk of exposure to PCDD/Fs in PM2.5 for children or adults is relatively low, there is a certain risk at some locations in winter, necessitating additional monitoring and control.
5. Genomic sequence analysis of a hantavirus found in Yunnan bat
Hongyan DENG ; Jinglin WANG ; Lili LI ; Yunyun XIN ; Mengmeng LIU ; Yun WANG ; Zhaojun DUAN
Chinese Journal of Experimental and Clinical Virology 2018;32(2):140-144
Objective:
To understand the novel bat-borne hantavirus on Yunnan province.
Methods:
Eighty-four bats were collected from Puer, Yunnan from July to August in 2016. A hantavirus, named as DodeHV, was identified in Pomona Roundleaf Bat by high-throughput sequencing technology. Nested or Semi-nested reverse transcription-polymerase chain reaction was used to confirm and screen DodeHV, as well as to amplify its genome sequence by specific primers. Then homology and phylogenetic analysis of all three segments were conducted by using software MegAlign and MEGA 6.0.
Results:
All the positive individuals are Pomona Roundleaf bats by PCR in this study and 4 out of 67 (5.97%) Pomona Roundleaf bats were positive. The complete ORF of DodeHV S Segment, the most sequence of M and the complete sequence of L segment were obtained. It shared the highest homology with the strain XSV-VN1982B4 found on Phu Tho Province, Viet- Nam in 2013, the nucleotide sequence identities of S, M, L segment compared with XSV was 79.0%, 79.2%, 79.9% respectively, and its amino acid sequence identities was 93.4%, 94.8%, 96.6% respectively. Meanwhile, phylogenetic analysis showed that DodeHV was also closely related to strain XSV-VN1982B4.
Conclusions
The discovery of DodeHV enriched the virus reservoir of our country which is meaningful to public health. It is indicated that there is potential risk of bat-borne DodeHV at cross-border infection at frontier.
6.Methcathinone Increases Visually-evoked Neuronal Activity and Enhances Sensory Processing Efficiency in Mice.
Jun ZHOU ; Wen DENG ; Chen CHEN ; Junya KANG ; Xiaodan YANG ; Zhaojuan DOU ; Jiancheng WU ; Quancong LI ; Man JIANG ; Man LIANG ; Yunyun HAN
Neuroscience Bulletin 2023;39(4):602-616
Methcathinone (MCAT) belongs to the designer drugs called synthetic cathinones, which are abused worldwide for recreational purposes. It has strong stimulant effects, including enhanced euphoria, sensation, alertness, and empathy. However, little is known about how MCAT modulates neuronal activity in vivo. Here, we evaluated the effect of MCAT on neuronal activity with a series of functional approaches. C-Fos immunostaining showed that MCAT increased the number of activated neurons by 6-fold, especially in sensory and motor cortices, striatum, and midbrain motor nuclei. In vivo single-unit recording and two-photon Ca2+ imaging revealed that a large proportion of neurons increased spiking activity upon MCAT administration. Notably, MCAT induced a strong de-correlation of population activity and increased trial-to-trial reliability, specifically during a natural movie stimulus. It improved the information-processing efficiency by enhancing the single-neuron coding capacity, suggesting a cortical network mechanism of the enhanced perception produced by psychoactive stimulants.
Mice
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Animals
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Reproducibility of Results
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Neurons
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Sensation
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Perception