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Photohardening therapy, also known as photodesensitization therapy, refers to the phototherapy and photochemotherapy of idiopathic actinic dermatoses, and its goal is to improve the patients′ tolerance to sunlight and prevent disease flares. Its mechanisms of action involve a variety of cellular and inflammatory factors. This therapy is suitable for all idiopathic actinic dermatoses, with definite efficacy and good safety. However, the treatment specificity usually leads to poor compliance. The development of UVA1 rush hardening and home phototherapy is expected to solve this problem.
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Objective:To investigate serum lipidomic profiles in patients with chronic actinic dermatitis (CAD), and to search for biomarkers of CAD.Methods:A retrospective analysis was conducted. Serum samples were collected from 46 patients with CAD and 16 age- and gender-matched healthy controls in the Guangzhou Institute of Dermatology from April 2011 to December 2021. Changes in serum lipid composition and expression were assessed by liquid chromatography-mass spectrometry. Principal component analysis, partial least squares discriminant analysis, and orthogonal partial least squares discriminant analysis were performed to screen differential biomarkers, and receiver operating characteristic (ROC) curve analysis was conducted to screen diagnostic markers. Comparisons of the age and gender distribution between groups were performed using t test and chi-square test, respectively. Results:The 46 CAD patients were aged from 30 to 84 (60.39 ± 10.52) years, including 41 males and 5 females; the 16 healthy controls were aged from 50 to 89 (59.81 ± 10.72) years, including 14 males and 2 females; there were no significant differences in the age or gender distribution between the two groups (age: t = 0.19, P = 0.853; gender: χ2 = 0.03, P = 0.859). Totally, 4 136 lipid molecules belonging to 40 subclasses were identified in the serum samples from CAD patients as well as healthy controls. Twenty-two differential lipid molecules were identified between the CAD patients and healthy controls, belonging to 9 subclasses (triglycerides, sphingomyelin, phosphatidylserine, phosphatidylethanolamine, monofatty acid glycerides, lysophosphatidylcholine, hexose ceramide, diglycerides, and cardiolipin). When the combinations of triglycerides (37.7e) and Na, those of monoglycerides (22.3) and NH 4, or those of phosphatidylserine (18.0_18.1) and H served as diagnostic markers separately, the areas under the ROC curve (AUCs) were all > 0.8, and the AUCs of 16 differential lipid molecules were all > 0.7. Conclusion:The serum lipid composition differed between healthy controls and CAD patients, and the combinations of triglycerides (37.7e) and Na, those of monoglycerides (22.3) and NH 4, and those of phosphatidylserine (18.0_18.1) and H may be promising biomarkers for the diagnosis of CAD.
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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.
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Objective:To ascertain the distribution of interventional radiodiagnosis and treatment resources and the current situation of radiological protection in Beijing, standardize the interventional radiodiagnosis and treatment, and promote the implementation of radiation protection regulatory measures.Methods:Various medical institutions at differetn levels that perform interventional radiodiagnosis and treatment in Beijing were identified as the survey objects. With special questionnaires designed, the investigation groups at two levels of municipality and district was established to investigate the basic situation of interventional radiology and occupational health monitoring by the end of 2020. The indexes and parameters were analyzed and evaluated under the relevant national regulations and standards.Results:By the end of 2020, there were 93 medical institutions performing interventional radiology in Beijing, together with 236 digital subtraction angiography machines (DSA) with higher than 800 mA. A total of 135 593 cases of interventional radiological surgical operation were performed. There were 40 hospitals annually performing more than 1 000 cases and 41 hospitals perfoming 10-1 000 cases. There were 3 539 interventional radiological workers, with 99.0% holding radiological worker certificates. The passing rates of occupational health examination, personal dose monitoring and radiation protection training were 96.9%, 99.5% and 95.8%, respectively. A total of 3 532 sets of protective articles were provided for the workers, of which 98.9% were equipped with split or integrated lead clothing, but 6.5% were not equipped with lead protective glasses and 54.9% were not equipped with lead protective gloves.Conclusions:The radiation protection management for the interventional radiodiagnosis and treatment is generally good. However, the regulatory mechanism should be further improved based on the current distribution of interventional radiodiagnosis and treatment resources, with focus on strengthening the occupational health examination, the radiation protection training, and the configuration and use of protective equipment.
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Objective:To evaluate the efficacy for nanoscale microneedle injection of compound betamethasone combined with 308 nm excimer laser in the treatment of stable vitiligo patients.Methods:A total of 80 patients with stable vitiligo were enrolled in Guangzhou Dermatology Hospital from May 2018 to May 2020. There were 40 patients (21 males and 19 females) in control group, aged 17-65 (32.4±1.7) years, and 40 patients (20 males and 20 females) in observation group, aged 18-67 (28.7±1.8) years. The control group was treated with compound betamethasone injection packet combined with 308 nm excimer laser. The observation group was treated with nanoneedle injection of compound betamethasone combined with 308 nm excimer laser. We compared the clinical efficacy and incidence of adverse reactions between the two groups.Results:Comparison of clinical efficacy showed that after 3 months of treatment, the total effective rates of the observation group and the control group were 80.00% and 67.50%, respectively, with significant difference (χ 2=4.560, P<0.05). After 3 months of treatment, the white spot area of the control group was (9.89±1.65) cm 2, which was significantly higher than that of the observation group (7.83±1.78) cm 2 ( t=5.370, P<0.05). Conclusions:The nanoneedle injection of compound betamethasone combined with 308 nm excimer laser in the treatment of stable vitiligo is effective and safe.
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Objective:To analyze the risk factors of postinflammatory hyperpigmentation (PIH) after laser in the treatment of acquired bilateral nevus of Ota-like macules (ABNOM).Methods:A retrospective study was conducted to follow up 120 patients with acquired bilateral nevus of Ota-like macules in the Department of Laser and Physiotherapy, Guangzhou Institute of Dermatology between January 2011 and December 2018, which accepted 1064-nm Q-switched neodymium: yttrium-aluminum-garnet laser treatment. The difference was analyzed between different age, sex, clinical classification, Fitzpatrick skin classification, ABNOM with melasma and postinflammatory pigmentation after laser treatment. Logistic regression was used to analyze the risk factors of postinflammatory hyperpigmentation after 1064-nm Q-switched neodymium: yttrium-aluminum-garnet laser treatment of acquired bilateral nevus of Ota-like macules.Results:Fifty-three ABNOM patients (44.17%) developed PIH after laser treatment. Univariate analysis showed that age, clinical classification, Fitzpatrick skin classification and the patients with both ABNOM and melasma all affected the occurrence of PIH after laser in the treatment of ABNOM, and the difference was statistically significant ( P<0.01). Logistic regression showed that older age, more severe clinical classification and the presence of ABNOM with melasma were the risk factors of PIH after treatment of ABNOM. Conclusions:ABNOM patients should be treated as early as possible. The risk of inducing PIH is great after laser treatment in patients with more severe clinical classification and patients with both ABNOM and melasma.
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Objective:To compare the detection rate of genital Chlamydia trachomatis (CT) DNA between urine and urethral/cervical swab samples. Methods:From December 2018 to December 2019, a total of 1 475 outpatients were collected from sexually transmitted disease clinics in 7 medical institutions, such as Department of Venereology, Guangzhou Institute of Dermatology, including 1 118 males and 357 females. One urethral/cervical swab sample and one urine sample were collected successively from each patient. Real-time fluorescence-based PCR was performed to detect CT DNA in urine and urethral/cervical swab samples, and paired chi-square test was used to compare the positive rate of CT DNA between the 2 kinds of samples. Random- or fixed-effect meta-analysis was conducted for the test of heterogeneity and merging of positive rates of CT DNA in the urine and urethral/cervical swabs among 7 medical institutions.Results:The positive rate of CT DNA in the urine samples was significantly higher than that in the swab samples from 4 medical institutions (all P < 0.05) , while there was no significant difference in the positive rate of CT DNA between the 2 kinds of samples from 3 medical institutions (all P > 0.05) . The heterogeneity ( I2) estimates of the CT-DNA positive rate in urine and swab samples among different medical institutions were 78.6% (95% CI: 55.9% - 89.6%) and 73.7% (95% CI: 43.7% - 87.7%) , respectively; meta-analysis showed that the total merged positive rate of CT DNA in the urine samples was 10.8% (95% CI: 7.2% - 15.9%) , which was significantly higher than that in the swab samples (7.8%, 95% CI: 4.9% - 12.1%; χ2 = 39.2, P < 0.05) . Compared with the swab sample-based CT-DNA detection method, the sensitivity, specificity, positive predictive value, negative predictive value and consistency rate of the urine sample-based CT-DNA detection method were 97.0% (128/132) , 96.3% (1 293/1 343) , 71.9% (128/178) , 99.7% (1 293/1 297) , and 96.3% (1 421/1 475) , respectively. The positive rate of CT DNA in the urine samples from 1 118 male patients was 11.0% (95% CI: 7.2% - 16.5%) , which was significantly higher than that in the swab samples (7.6%, 95% CI: 4.9% - 11.8%; χ2 = 34.3, P < 0.05) . There was no significant difference in the positive rate of CT DNA between the urine (11.9%, 95% CI: 7.7% - 17.9%) and cervical swab samples from 357 female patients (10.4%, 95% CI: 7.6% - 14.0%; χ2 = 3.2, P > 0.05) . Conclusions:The positive rate of CT DNA in urine samples is higher than or similar to that in urethral/cervical swab samples. The urine sample-based CT-DNA detection method has characteristics of convenience, non-invasiveness, painlessness and low cost, and is worthy of clinical promotion.
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Objective:To investigate the effect of pterostilbene on the growth, apoptosis and autophagy of a human papillomavirus type 16 (HPV-16) -immortalized cervical epithelial cell line H8.Methods:H8 cells were treated with pterostilbene at different concentrations of 0 (control group) , 25, 50, 75, 100 μmol/L for 24 and 48 hours. Cell counting kit-8 (CCK8) assay was performed to evaluate the cellular proliferative activity, flow cytometry was conducted to detect apoptosis and cell cycle, monodansylcadaverine (MDC) staining and fluorescence microscopy were performed to detect autophagy, and Western blot analysis was conducted to determine the expression of the cell cycle-related protein cyclinD1, apoptosis-related proteins caspase-3 and caspase-9, autophagy-related proteins Beclin1, microtubule-associated protein 1 light chain 3 (LC3) -Ⅱ/Ⅰ, ATG5 and P62, as well as HPV oncoproteins E6 and E7. Statistical analysis was carried out by using one-way analysis of variance, repeated measures analysis of variance and least significant difference- t test. Results:After 48-hour treatment with pterostilbene at different concentrations of 0, 25, 50, 75, 100 μmol/L, the relative cellular proliferation rate significantly differed among the groups (100.00% ± 1.56%, 99.02% ± 4.97%, 93.59% ± 2.01%, 81.28% ± 4.90%, 69.17% ± 7.56%, respectively; F = 77.22, P < 0.05) , and gradually decreased along with the increase in the concentration of pterostilbene; compared with the control group, the pterostilbene groups all showed significantly decreased cellular proliferation rate (all P < 0.05) . After 24-hour treatment with pterostilbene, the proportions of H8 cells at G1, G2 and S phases significantly differed among the above groups ( F = 7 845.00, 51.14, 266.50, respectively, all P < 0.05) ; compared with the control group, the pterostilbene groups showed significantly increased proportions of H8 cells at G1 and G2 phases (all P < 0.05) , but significantly decreased proportions of H8 cells at S phase ( P < 0.05) . After 48-hour treatment with pterostilbene, the apoptosis rate was significantly higher in the 25-, 50-, 75- and 100-μmol/L pterostilbene groups (14.66% ± 0.22%, 13.50% ± 0.49%, 14.56% ± 0.19%, 15.30% ± 0.76%, respectively) than in the control group (11.58% ± 0.50%, all P < 0.05) . After 24-hour treatment with pterostilbene, MDC staining showed only a small number of H8 cells with bright dot-like fluorescence in the control group, but increased number of autophagosome-positive H8 cells with bright dot-like fluorescence in the pterostilbene groups. Western blot analysis revealed that there were significant differences in the protein expression of cyclin D1, caspase-3, caspase-9, Beclin1, LC3-Ⅱ/Ⅰ, ATG5, P62, E6 and E7 among the control and pterostilbene groups after 24- and 48-hour treatment with pterostilbene (all P < 0.05) . The treatment with pterostilbene could down-regulate the expression of cyclin D1, E6 and E7, and up-regulate the expression of caspase-3, caspase-9, Beclin1, LC3-Ⅱ/Ⅰ, ATG5 and P62, with significant differences between the control group and most pterostilbene groups in expression of the above proteins (all P < 0.05) . Conclusion:Pterostilbene can inhibit the proliferation of H8 cells, promote their apoptosis and autophagy, and down-regulate the expression of oncogenes E6 and E7.
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Objective:To explore the differential expression of Toll-like receptor 7/9 and myeloid differentiation factor 88 (MyD88) in skin lesions between patients with vitiligo and healthy individuals as well as their clinical significance.Methods:We collected vitiligo patients in the Guangzhou Institute of Dermatology and Venerology from June 2016 to June 2018. There were 24 vitiligo patients, 12 males and 12 females, aged 5 to 65 (28.75±13.12) years, with medical history of 14 days to 20 years. The expression levels of TLR7/9 and MyD88 in skin lesions were determined from 24 patients with vitiligo and 20 healthy controls by immunohistochemistry.Results:The expression levels of TLR7 and MyD88 in skin lesions were significantly higher in patients with vitiligo than that in the controls, and the difference was statistically significant. The expression level of TLR9 in skin lesions was significantly higher in patients with vitiligo than that in the controls, while no significant difference was observed between the two groups.Conclusions:The expression level of TLR7 and TLR9 is elevated in skin lesions from patients with vitiligo. The abnormal expression level of TLR7 and TLR9 possibly participates in the pathogenesis of vitiligo.
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Objective To determine the expression of miRNA-148a-3p in CD4+ T lymphocytes in peripheral blood of patients with psoriasis vulgaris,and to explore its role in occurrence of psoriasis vulgaris.Methods Totally,20 patients with psoriasis vulgaris and 20 healthy controls were enrolled from Guangzhou Institute of Dermatology between July 2017 and April 2018.Peripheral venous blood samples were obtained from these subjects,and CD4+ T lymphocytes were isolated from these peripheral blood samples by magnetic cell sorting system.Real-time quantitative PCR (RT-PCR) was performed to determine the expression of miRNA-148a-3p in CD4+ T lymphocytes in the peripheral blood.Potential target genes of miRNA-148a were predicted by using bioinformatics software,and verified by using a dual-luciferase reporter system.Western blot analysis was conducted to determine the protein expression of Bcl-2 interacting mediator of cell death (Bim,the potential target gene of miRNA-148a-3p) in the CD4+ T lymphocytes of the subjects.Statistical analysis was carried out with SPSS 20 software by two sample-t test for comparing the means of normally distributed data,and by Pearson correlation analysis for analyzing the correlation of two variables.If the data were not normally distributed,Mann Whitney U test was used for comparing means between two groups,and Spearman correlation analysis for analyzing the correlation of two variables.Results The miRNA-148a-3p expression in the CD4+ T lymphocytesin the psoriasis vulgaris group (18 cases,5.61 ± 1.66) was significantly higher than that in the healthy control group (12 cases,1.00 ± 0.26;U =12,P ＜ 0.05),and was positively correlated with the psoriasis area severity index (PASI) score (r =0.93,P ＜ 0.001).Bim was predicted to be one of the potential target genes of miRNA-148a-3p by bioinformatics software,which was also verified by using a dual-luciferase reporter system.The protein expression of Bim in the CD4 + T lymphocytes was significantly lower in the psoriasis vulgaris group (11 cases,0.69 ± 0.07) than in the healthy control group (8 cases,0.93 ± 0.06;t =4.38,P ＜ 0.01),and the protein expression of Bim in the patients with psoriasis vulgaris was negatively correlated with PASI score (r =-0.774,P ＜ 0.01).Conclusion miRNA-148a-3p is overexpressed in CD4+ T cells in the peripheral blood of patients with psoriasis vulgaris,which may regulate the protein expression of Bim,leading to abnormal activation of CD4+ T cells,and then participate in the occurrence and development of psoriasis.
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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.
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Objective To explore the effect of tea polyphenols on the growth of human papillomavirus 16 (HPV16) subgenes-immortalized human cervical epithelial cells (H8 cells).Methods Cultured H8 cells were divided into 5 groups to be treated with 0 (control group),6.25,12.5,25 and 50 mg/L tea polyphenols respectively for 24,36,and 48 hours,and then cell counting kit-8 (CCK8)assay was performed to detect cell proliferation.After 24 hours of incubation,flow cytometry was conducted to detect cell apoptosis and cell cycle,and fluorescence microscopy to observe the morphology of apoptotic cells.Results After incubation with tea polyphenols at different concentrations for 24,36 and 48 hours,the proliferation of H8 cells was inhibited,and 12.5 mg/L tea polyphenols could inhibit the relative growth rate of H8 cells in a time-dependent manner.Flow cytometry showed that there was a significant difference in cell apoptosis rate among the 6.25-,12.5-,25-,50-mg/L tea polyphenols groups and the control group (52.62％ ± 0.62％,52.22％ ± 0.72％,42.52％ ± 0.90％,45.96％ ± 2.11％,29.96％ ± 0.70％ respectively,F =272.0,P ＜ 0.05).Moreover,all the tea polyphenol groups showed significantly increased cell apoptosis rate compared with the control group (all P ＜ 0.05).Fluorescence microscopy showed karyopyknosis,nuclear fragmentation and other typical apoptotic morphological changes in H8 cells in tea polyphenols groups.There were significant differences in the percentage of cells in G1,G2 phase and cell proliferation index among the 5 groups (all P ＜ 0.05).Compared with the control group,the 6.25-,12.5-,25-mg/L tea polyphenols groups showed significantly increased percentage of cells in G1 phase (55.96％ ± 0.72％,54.12％ ± 3.20％,65.30％ ± 1.51％ respectively,all P ＜ 0.05),but significantly decreased percentage of cells in G2 phase (3.17 ± 1.82％,4.94 ± 1.46％,4.65 ± 4.26％ respectively,all P ＜ 0.05) and lower cell proliferation index(0.44 ± 0.01,0.46 ± 0.02,0.36 ± 0.01 respectively,all P ＜ 0.05).Conclusion Tea polyphenols can inhibit the proliferation of H8 cells,induce cell apoptosis,and block cell cycle progression.
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OBJECTIVE:To observe clinical efficacy and safety of salmeterol fluticasone combined with tiotropium bromide in the treatment of COPD via different inhalation devices.METHODS:Eighty COPD patients were selected from our hospital during Jan.2014 to Jan.2015,and then divided into trial group and control group according to random number table,with 40 cases in each group.Both groups were given Salmeterol fluticasone inhalant 500 μg,bid+Tiotropium bromide inhalant 18 μg,qd.Control group was given medicine via inhalation device coming with medicine,while trial group was given medicine via gas compression type ultrasonic spray inhalator.Both groups were treated for 1 year.Blood concentration of medicine 0.5 h after medication,mMRC score and COPD acsessment test (CAT) score 3,6,9 months after treatment,the times of acute exacerbation during treatment,FEV1％ before and af ter treatment were all observed in 2 groups.The occurrence of ADR was recorded.RESULTS:Four cases withdrew from trial group and 1 case from control group.After medication,there was no statistical significance in blood concentration of fluticasone,salmeterol and tiotropium bromide between 2 groups (P＞0.05).0.5 h after medication,mMRC score of trial group was slightly lower than that of control group,without statistical significance (P＞0.05);CAT score of it was significantly lower than that of control group,with statistical significance (P＜0.05).The times of acute exacerbation in trial group during treatment was significantly less than control group,with statistical significance (P＜0.05).The decrease of FEV1％ in trial group was slightly lower than control group,without statistical significance (P＞0.05).The incidence of ADR in trial group was significantly lower than control group,with statistical significance (P＜0.05).CONCLUSIONS:For COPD patients,salmeterol fluticasone combined with tiotropium bromide via gas compression type ultrasonic spray inhalator is better than inhalation device coming with medicine in clinical efficacy and safety.
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Objective To explore the methods for quality management and continuous improvement of nursing care quality in the orthopedic demonstration ward by taking the hospital accreditation as an opportunity. Methods From July 2012 to June 2013, the continuous care quality improvement in the ward was carried out to find out the problems with PDCA (plan, do, check, action) cycle method, including enhancing the function of orthopedic nursing quality management groups, conducting all-staff training and improving the knowing rate by referring to the standards of hospital assessment standards. Results After the performance of whole-process quality management, the percentage of indexes assessed at level A, B and C was increased from 42.2%to 50.0%, 17.2%to 14.7%and 40.2%to 35.3%, respectively. The score of nurses' responsibility accreditation was increased from 92 to 95. The rates of patient and nursing staff satisfaction were increased from 91.8%to 98.9%and 92.57%to 97.7%, respectively. Conclusion In accordance with the standards for hospital accreditation, the continuous improvement of nursing quality in the orthopedic demonstration wards can improve the specialist care of orthopedic care, improve patients' and nurses' satisfaction, thus making the daily work more scientific and standardized.
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Objective To evaluate effects of tea polyphenols on the mRNA and nucleoprotein expression of Nrf2/Bach1 in human skin fibroblasts (HSFs).Methods Some HSFs were incubated with tea polyphenols at different concentrations of 0,2.5,5,10,20 and 40 mg/L for 24 hours.Methyl thiazolyl tetrazolium (MTT) assay was conducted to evaluate the proliferative activity of HSFs to screen the optimal concentration of tea polyphenols.Then,some other HSFs were treated with tea polyphenols at this optimal concentration for 24 hours.Real-time quantitative PCR (RT-qPCR) was performed to determine mRNA expression of Nrf2 and Bach1,Western blot analysis to measure nuclear expression of Nrf2 and Bach1 proteins,and immunofluorescence assay to determine the distribution of Nrf2 and Bach1 protein in the cell nucleus.Results MTT assay showed that 5 mg/L tea polyphenols had no obvious effects on the proliferation of HSFs,so 5 mg/L was chosen as the optimal concentration of tea polyphenols for subsequent experiments.HSFs cultured without tea polyphenols served as control group.After the treatment,the 5-mg/L tea polyphenol group showed significantly decreased mRNA and nuclear protein expression of Bach 1 (mRNA:0.629 ± 0.077 vs.0.940 ± 0.033,t =6.397,P ＜ 0.05;protein:1.424 ± 0.171 vs.16.966 ± 1.702,t =15.730,P ＜ 0.05),but significantly increased mRNA and nuclear protein expression of Nrf2 (mRNA:1.467 ± 0.076 vs.0.977 ± 0.091,t =7.133,P ＜ 0.05;protein:6.929 ± 0.121 vs.3.537 ± 0.126,t =33.636,P ＜ 0.05) compared with the control group.Immunofluorescence assay showed increased accumulation of Nrf2 protein,but decreased accumulation of Bach1 protein in the nucleus.Conclusion Tea polyphenols can promote the mRNA and nuclear protein expression as well as nuclear distribution of Nrf2,but suppress the mRNA and nuclear protein expression as well as nuclear distribution of Bach 1,finally exerting antioxidative effects.
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Objective To investigate the protective effect of lycium barbarum polysaccharide (LBP) on DNA damage of HSF cells induced by UV.Methods We established the model of UV induced photo damage in HSF cells.We detected the viability of HSF cells by using MTT colorimetry.The UV absorption spectrum of LBP was also measured by UV spectrophotometer.The level of ROS was detected by DCFH-DA fluorescent probe method.Comet assay was employed to evaluate the DNA strand breakage damage.Results When the concentration of LBP was less than or equal to 300μg/ml,there was no significant effect on the proliferation of HSF cells (P＞0.05).When the concentration was more than 300 μg/ml,it could inhibit the cell proliferative activities (P＜0.05).Compared to the UV groups,UV+LBP groups can respectively improve the cell proliferation activity (P＜0.05).The absorbance was slight range 280 from 400 nm.Compared with the UV group,the relative fluorescence intensity and the migration distance of UV+ LBP groups were significantly decreased (P＜0.05).Conclusions Lycium barbarum polysaccharide can effectively inhibit the proliferation activity and protect the breakage of DNA strand induced by UV,which is probably due to its action of removing free radicals.
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Objective To evaluate the scavenging effect of crude polysaccharides extracted from Lycium barbarum (LBP) on reactive oxygen species in ultraviolet radiation-induced HaCaT cells,and to explore its possible mechanism.Methods Cultured immortalized human keratiuocyte HaCaT cells were divided into 6 groups:blank control group receiving no treatment,LBP group treated with crude LBP alone,ultraviolet A (UVA) group treated with UVA radiation alone,ultraviolet B (UVB) group treated with UVB radiation alone,UVA + LBP group treated with crude LBP for 24 hours followed by UVA radiation,and UVB + LBP group treated with crude LBP for 24 hours followed by UVB radiation.MTT colorimetry was performed to evaluate the cellular proliferative activity,UV spectrophotometric method to measure the UVA and UVB absorption of crude LBP,dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe assay to detect the level of ROS,enzymatic-biochemical method to estimate the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px),as well as to detect the leakage of lactate dehydrogenase (LDH).Results Crude LBP at different concentrations of 0,100,200,300,400,500,600,1 500,2 000 mg/L had no obvious effects on the proliferative activity of HaCaT cells.Crude LBP had a high transmittance of ultraviolet rays at 280-400 nm.Compared with the blank control group,the UVA group and UVB group both showed significantly higher LDH leakage and ROS level,lower activities of SOD and GSH-Px (P ＜ 0.001 or 0.05).Pretreatment with crude LBP before the ultraviolet radiation could significantly increase the activities of SOD and GSH-Px,decrease the LDH leakage and ROS level in the UVA + LBP group and UVB + LBP group compared with the UVA group or UVB group (P ＜ 0.05).Conclusion Crude LBP have no effect of sunscreening agents,but can effectively scavenge ROS,decrease LDH leakage,inhibit ultraviolet radiation-induced photodamage in HaCaT cells,which may be associated with the enhancement of antioxidant enzyme activity.
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Objective To explore the efficacy of cold packs combined with ice compress in treatment of pain after erbium fractional photothermal therapy on acne scars. Methods Eighty cases which were confirmed to the criteria were randomized into two groups:treatment group and control group. The treatment group (n=40) treated with cold packs combined with cold icy compress immediately after the surgery for 30~40 mins. The control group (n=40) was given icy compress therapy immediately after the surgery for 30~40 mins. The therapy continued for three days after the surgery on the two groups. The self-feeling symptom and pain relieving time were compared between the two groups. Result The time for pain relief in the wounds and the time for scabbing were both significantly shorter than those in the control group (P<0.01). Conclusion The cold packs combined with ice compress therapy can relieve the pains effectively and it can shorten the recovery time after fractional photothermal therapy.
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The immunological characterization in the pathogenesis of urticaria, mainly regarding cytokine profile, needs more investigation. In this study, subgroups of the T, B and natural killer (NK) lymphocyte from peripheral blood and serum levels of interleukin (IL)-15, IL-21 and immunoglobulin (Ig)E were examined in patients with acute urticaria (AU) and chronic urticaria (CU). Moreover, symptom scores and course of the patients were assessed. The percentage of NK cells and the ratio of CD4(+)/CD8(+) increased, however, CD8(+) decreased in CU compared to controls (P < 0.01). But no significant changes of T, B and NK lymphocyte were found in AU. IL-15 and IL-21 significantly decreased in AU and CU, but IgE increased. CU with a positive autologous serum skin test were more likely to be associated with longer course and higher CD3(+), B cells and IL-21, and lower IgE (P < 0.01). Weak negative correlations were demonstrated between CD3(+), CD8(+) and scores in CU (r = -0.23, -0.25, P < 0.05). Significant correlations were found between B cells and scores and course in CU (r = 0.49, 0.65, P < 0.01). Moreover, a significant correlation was found between IL-21 and IgE (r = 0.42, P < 0.01) in CU. But no significant correlations were found in AU. Our findings supported the concept that both humoral immunity and cellular immunity dysregulation in the pathogenesis of urticaria - mainly related to the decrease of the serum levels of IL-15 and IL-21 - may induce the increasing expression of IgE produced by B cells.