1.Technical Research of Non-contact Electrocardiogram Based on Capacitive Coupling.
Shan GAO ; Yongqin LI ; Liang WEI ; Jie XIONG ; Dacheng LIAO ; Bihua CHEN
Journal of Biomedical Engineering 2015;32(3):553-557
Based on the capacitance coupling principle, we studied a capacitive way of non-contact electrocardiogram (EGG) monitoring, making it possible to obtain ECG on the condition that a patient is habilimented. Conductive fabric with a good electrical conductivity was used as electrodes. The electrodes fixed on a bed sheet is presented in this paper. A capacitance comes into being as long as the body gets close to the surface of electrode, sandwiching the cotton cushion, which acts as dielectric. The surface potential generated by heart is coupled to electrodes through the capacitance. After being processed, the signal is suitable for monitoring. The test results show that 93.5% of R wave could be detected for 9 volunteers and ECG with good signal quality could be acquired for 2 burnt patients. Non-contact ECG is harmless to skin, and it has advantages for those patients to whom stickup electrodes are not suitable. On the other hand, it is convenient to use and good for permanent monitoring.
Electric Conductivity
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Electrocardiography
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instrumentation
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Electrodes
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Humans
2.Effect of pterostilbene on the expression and activity of antioxidant enzymes in ultraviolet B-radiated HaCaT Cells
Huiyan DENG ; Huaping LI ; Quan CHEN ; Runxiang LI ; Bihua LIANG ; Aili GAO ; Xin ZHOU ; Huilan ZHU
Chinese Journal of Dermatology 2018;51(4):274-278
Objective To evaluate the protective effect of pterostilbene against ultraviolet B (UVB)-induced acute damage in HaCaT cells,and to explore related mechanisms.Methods The 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazo1ium (MTS) assay and flow cytometry were performed to estimate the proliferative activity and the apoptosis and necrosis rate of HaCaT cells treated with different concentrations of pterostilbene respectively,so as to screen the non-toxic concentration of pterostilbene.HaCaT cells were randomly divided into several groups:normal control group receiving no treatment,UVB group irradiated with 57 mJ/cm2 UVB,3 pterostilbene groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours,3 pterostilbene + UVB groups treated with 2.44,4.88 and 9.75 μmol/L pterostilbene respectively for 24 hours followed by UVB radiation.Western blot analysis was conducted to detect changes of the transcription factor NF-E2-related factor 2 (Nrf2) expression in cell nuclei and cytoplasm before and after the treatment with pterostilbene and UVB,quantitative PCR to determine the mRNA expression of catalase and superoxide dismutase in the HaCaT cells,and enzyme-linked immunosorbent assay (ELISA) to evaluate the activity of catalase and superoxide dismutase.Results MTS assay and flow cytometry showed that 2.44,4.88 and 9.75 μmol/L pterostilbene had non-toxic effect on HaCaT cells.The protein expression of Nrf2 in the nuclei and cytoplasm in the normal control group was 1.03 ± 0.08 and 1.04 ± 0.11 respectively.Compared with the normal control group,the protein expression of Nrf2 in the nuclei and cytoplasm experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups,and the UVB group showed similar protein expression of Nrf2 in the cytoplasm,but significantly increased protein expression of Nrf2 in the nuclei (1.77 ± 0.08,q =17.24,P < 0.01).Compared with the normal control group and UVB group,the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups all showed significantly lower protein expression of Nrf2 in the cytoplasm (0.86 ± 0.10,0.87 ± 0.11 and 0.46 ± 0.11 respectively,all P < 0.05),but significantly higher protein expression of Nrf2 in the nuclei (2.38 ± 0.11,2.57 ± 0.11 and 2.07 ± 0.13,all P < 0.01).As qPCR showed,UVB radiation could significantly inhibit the mRNA expression of CAT (P < 0.05),but had no obvious effect on the mRNA expression of SOD (P > 0.05).The mRNA expression of CAT and SOD experienced no significant changes in the 2.44-,4.88-and 9.75-μmol/L pterostilbene groups compared with the normal control group (P > 0.05).However,2.44,4.88 and 9.75 μmol/L pterostilbene could significantly reduce the inhibitory effect of UVB radiation on the mRNA expression of CAT (P < 0.05) and up-regulate the mRNA expression of SOD in the pterostilbene + UVB groups (P < 0.05).ELISA revealed that UVB radiation could inhibit the activity of CAT and SOD in the HaCaT cells (both P < 0.001),while 2.44,4.88 and 9.75 μmol/L pterostilbene could reduce the inhibitory effect of UVB radiation on the activity of CAT and SOD (all P < 0.05).However,the activity of CAT and SOD were still lower in the 2.44-,4.88-and 9.75-μmol/L pterostilbene + UVB groups than in the normal control group (P < 0.05).Conclusion Pterostilbene can prevent UVB-induced acute damage in HaCaT cells by activating the Nrf2 pathway and up-regulating the expression of the downstream antioxidant enzymes CAT and SOD.
3.Dynamic changes of peripheral blood T lymphocytes in COVID-19 patients
Wanling CHEN ; Weifeng WANG ; Wentao LI ; Fengmei CHEN ; Bihua GAO ; Feng LONG ; Houyang ZENG ; Jiannan LYU
Chinese Journal of Microbiology and Immunology 2020;40(7):495-498
Objective:To analyze the dynamic changes of T lymphocytes in patients with COVID-19.Methods:Blood samples were collected from 40 COVID-19 cases and 40 healthy controls in Beihai People′s Hospital from January to February, 2020. The counts of CD4 + T and CD8 + T lymphocytes were detected by flow cytometry. Moreover, the T lymphocyte counts in 24 convalescent patients with two consecutive negative nucleic acid test results were also detected. Results:The leukocytes and lymphocytes in the patients with acute COVID-19 were significantly lower than those in the healthy controls [(4.71±1.54)×10 9 cell/L vs (6.26±1.44)×10 9 cell/L, (1.13±0.41)×10 9 cell/L vs (1.51±0.39)×10 9 cell/L; both P<0.05]. The counts of CD4 + T and CD8 + T lymphocytes in the patients with acute COVID-19 were significantly lower than those in the healthy controls [(447.15±144.42) cell/μl vs (592.83±146.76) cell/μl, (309.35±173.05) cell/μl vs (397.20±136.94) cell/μl; both P<0.05], while no significant difference was observed in the CD4 + /CD8 + T cell ratio ( P>0.05). In the 24 convalescent COVID-19 patients, the counts of CD4 + T and CD8 + T lymphocytes were higher during convalescence than in the acute phase [(598.08±138.71) cell/μl vs (420.67±147.38) cell/μl, (439.08±166.94) cell/μl vs (296.67±151.06) cell/μl; both P<0.05], but there was no significant difference in the T lymphocyte counts between the convalescent patients and the healthy controls ( P>0.05). Conclusions:A transient immune deficiency occurred in patients with acute COVID-19, but the impaired immune function could restore to normal level during recovery.
4.Metformin inhibits ultraviolet A-induced photoaging of HaCaT cells by activating the adenosine monophosphate-activated protein kinase/nuclear factor-erythroid 2-related factor 2 signaling pathway
Huaping LI ; Aili GAO ; Bihua LIANG ; Huiyan DENG ; Jiaoquan CHEN ; Hui ZOU ; Tianyi LIN ; Sanquan ZHANG ; Huilan ZHU
Chinese Journal of Dermatology 2023;56(12):1123-1130
Objective:To evaluate the effect of metformin on ultraviolet A (UVA) -induced photoaging of an immortalized human keratinocytes cell line (HaCaT), and to explore its potential mechanisms.Methods:Cell counting kit 8 (CCK8) assay was performed to evaluate the effect of metformin at different concentrations (0 - 100 mmol/L) on the viability of HaCaT cells, and 10 mmol/L metformin was selected for subsequent experiments. Cultured HaCaT cells were divided into a blank control group (conventional culture), a metformin group (treated with culture medium containing 10 mmol/L metformin), a UVA irradiation group (conventional culture for 24 hours followed by 10 J/cm 2 UVA irradiation) and a metformin + UVA group (treated with culture medium containing 10 mmol/L metformin for 24 hours followed by 10 J/cm 2 UVA irradiation) ; UVA irradiation was performed at a dose of 10 J/cm 2 once a day for 3 consecutive days. After 4-day treatment, cells were collected, the β-galactosidase assay was performed to determine the proportion of senescent cells in each group, 2′, 7′-dichlorodihydrofluorescein diacetate assay to detect levels of intracellular reactive oxygen species (ROS), and the comet assay to detect DNA damage levels. Additionally, some HaCaT cells were divided into the blank control group, metformin group, 1.25 μmol/L dorsomorphin (an adenosine monophosphate-activated protein kinase [AMPK] inhibitor) + metformin group, and 2.5 μmol/L dorsomorphin + metformin group, and cells in the latter two groups were treated with 1.25 and 2.5 μmol/L dorsomorphin respectively for 2 hours, followed by the treatment with 10 mmol/L metformin for 24 hours. Western blot analysis was performed to determine the cellular localization and phosphorylation levels of nuclear factor-erythroid 2-related factor 2 (Nrf2). By using the small-interfering RNA (siRNA) -mediated silencing method, siRNA-Nrf2 was transfected into HaCaT cells to knock down Nrf2 expression (siRNA-Nrf2 group) ; 2.5 μmol/L dorsomorphin-treated HaCaT cells or Nrf2-knockdown HaCaT cells were treated with metformin and UVA irradiation (dorsomorphin + metformin + UVA group, siRNA-Nrf2 + metformin + UVA group, respectively), and the proportions of senescent cells were further calculated in each group. Statistical analysis was carried out by using one-way analysis of variance and two-way analysis of variance, and least significant difference (LSD) - t test was used for multiple comparisons. Results:Treatment with different concentrations of metformin for 24 hours could affect the viability of HaCaT cells to varying degrees ( F = 5 206.31, P < 0.001) ; there were no significant differences in the relative survival rates of HaCaT cells between the 10 - 20 mmol/L metformin groups and the control group (0 mmol/L metformin group, all P > 0.05), while the relative cell survival rates were significantly lower in the 25 - 100 mmol/L metformin groups than in the control group (all P < 0.05). After UVA irradiation, HaCaT cells shrank significantly and became narrow and elongated, and the intercellular spaces increased; the relative cell survival rate was significantly lower in the UVA irradiation group (76.13% ± 1.03%) than in the blank control group (100.00% ± 1.24%, LSD- t = 14.86, P < 0.001), but significantly higher in the metformin + UVA group (106.69% ± 2.45%) than in the UVA irradiation group (LSD- t = 11.55, P < 0.001). Moreover, the UVA irradiation group showed significantly increased proportions of senescent cells (45.14% ± 4.98%), intracellular ROS levels (144.61% ± 4.91%), and percentages of DNA in the tail (75.33% ± 1.77%) compared with the blank control group (23.84% ± 1.89%, 55.49% ± 1.57%, 1.88% ± 0.29%, respectively, all P < 0.001), while the metformin + UVA group showed significantly decreased proportions of senescent cells (24.26% ± 1.34%), intracellular ROS levels (58.62% ± 2.17%), percentages of DNA in the tail (15.83% ± 1.23%) compared with the UVA irradiation group (all P < 0.001). Western blot analysis showed that the Nrf2 expression in the cytoplasm was lower in the 10 mmol/L metformin group than in the blank control group, while the phosphorylated Nrf2 expression in the nuclei was higher in the 10 mmol/L metformin group than in the blank control group, suggesting that metformin could effectively induce the phosphorylation of Nrf2 and its nuclear translocation; both the pretreatment with 1.25 and 2.5 μmol/L dorsomorphin could significantly reduce the phosphorylation levels of AMPKα and Nrf2 induced by 10 mmol/L metformin. The proportions of senescent cells in the dorsomorphin + metformin + UVA group and the siRNA-Nrf2 + metformin + UVA group were 67.84% ± 2.74% and 65.94% ± 1.33%, respectively, which were significantly higher than those in the metformin + UVA group (37.76% ± 1.64%, t = 14.45, 13.34, respectively, both P < 0.001) . Conclusion:Metformin may inhibit UVA-induced photoaging of HaCaT cells by activating the AMPK/Nrf2 signaling pathway, scavenging ROS and reducing DNA damage.