Objective To evaluate the effect of selenomethionine (Se-Met) against ultraviolet B (UVB)-induced oxidative damage to human HaCaT keratinocytes,and to explore its possible mechanisms.Methods Cultured HaCaT cells were divided into several groups:normal control group receiving no treatment,Se-Met groups treated with Se-Met at concentrations of 1,10,50,100,200 nmol/L and 1 μmol/L for 24 hours respectively,UVB groups irradiated with UVB of 30,60 and 90 mJ/cm2 respectively,Se-Met + UVB groups treated with Se-Met at concentrations of 1,10,50,100,200 nmol/L and 1 μmol/L for 24 hours firstly,then irradiated with UVB of 30,60 and 90 mJ/cm2 respectively.Subsequently,methyl thiazolyl tetrazolium (MTT) assay was performed to estimate cellular proliferative activity,flow cytometry to detect cell apoptosis,colorimetry to evaluate superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and to determine malondialdehyde (MDA) levels.Statistical analysis was carried out by using factorial design analysis of variance (ANOVA),one-way ANOVA and least significant difference (LSD) test.Results Factorial design ANOVA showed that UVB radiation had an inhibitory effect on the proliferative activity of HaCaT cells (F =128.04,P ＜ 0.05),which significantly decreased along with the increase of UVB doses,with significant differences between the three UVB groups (P ＜ 0.05).Se-Met pretreatment also affected cellular proliferative activity (F =5.95,P ＜ 0.05),which was significantly increased in Se-Met (10 nmol/L-1 μmol/L) + UVB groups compared with the UVB groups at corresponding doses (all P ＜ 0.05).There was no significant interaction effect on cellular proliferative activity between UVB radiation and Se-Met pretreatment (F =1.65,P ＞ 0.05).The apoptosis rate of HaCaT cells in the 30-mJ/cm2 UVB group was 31.9％ ± 2.67％,significantly higher than that in the normal control group (4.1％ ± 0.67％,P＜ 0.05) and in the 10-,50-,100-,200-nmol/L and 1-μmol/L Se-Met + 30-mJ/cm2 UVB groups (21.9％ ± 3.72％,17.2％ ± 1.67％,4.6％ ±-0.85％,7.5％ ± 1.86％ and 13.5％ ± 1.95％ respectively,all P ＜ 0.05).Similarly,SOD and GSH-Px activities were significantly weaker (both P ＜ 0.05),while MDA levels were higher (all P ＜ 0.05) in the 30-mJ/cm2 UVB group than in the normal control group;however,there was a significant increase in SOD and GSH-Px activities but a decrease in MDA levels in the Se-Met (10 nmol/L-1 μmol/L) + 30-mJ/cm2 UVB groups compared with the 30-mJ/cm2 UVB group (all P ＜ 0.05).Conclusions Se-Met can reduce UVB-induced oxidative damage to HaCaT cells,likely by enhancing antioxidase activity and decreasing oxygen radicals.

Objective To investigate the effect of euphorbiasteroid on inducing the apoptosis of HL-60 cells and demonstrate whether the Fas/FasL signaling pathway is involved in the induction of apoptosis. Methods HL-60 cells were treated with dose of 2.5,10,40 μg/ml of euphorbiasteroid in vitro for 24 h respectively.After that,cell counting Kit-8 was used to detect cell proliferation.The morphology of HL-60 cells were observed under light and fluorescent microscopy.The early cell apoptosis was detected by using flow cytometry with Annexin Ⅴ-FITC /PI double staining.The expressions of Fas,FasL,caspase-8 and caspase-3 mRNA were analyzed by the method of RT-PCR.The activities of caspase-8 and caspase-3 were examined by chromatometry.Results Compared with 1640 control group,HL-60 cell proliferation was inhibited significant-ly by euphorbiasteroid.The inhibition rates were (34.9 ±3.7)%,(54.6 ±5.2)% and (61.3 ±4.3)%respectively.Moreover,HL-60 cells exhibited typical morphological features.Early cell apoptosis rates of HL-60 cells were (23.4 ±3.1)%,(35.7 ±4.3)% and (53.2 ±3.9)% respectively.Furthermore,the expressions of Fas,FasL,caspase-3 and caspase-8 mRNA were up-regulated significantly after euphorbiasteroid administration in a dose-dependent manner (P<0.01 ).After treated with euphorbiasteroid,the activities of caspase-8 and caspase-3 were significantly enhanced (P<0.01 ).Conclusion The up-regulation effect of euphorbiasteroid on Fas/FasL signaling pathway might contribute to the apoptosis of HL-60 cells.

The combination of Angelicae sinensis Radix (Danggui, DG) and Astragali Radix (Huangqi, HQ) is a popular herb pair commonly used in clinic for the treatment of blood deficiency syndrome in China. The aim of this paper is to study the interaction of DG and HQ nourishing and tonifying blood effects by response surface method. The blood deficiency mice were induced by injecting N-acetylphenylhydrazine (sc) and cyclophosphamide (ip). The blood deficiency mice were administrated intragastrically with DG-HQ extracts (0:1, 1: 5, 2:5, 2:3, 1:1, 3:2, 5:2, 5:1, 1:0). The changes of the peripheral blood indexes and organ indexes were observed. The indexes were integrated by comprehensive index method; the interactions of DG and HQ were analyzed by the response surface diagram established with Matlab software. The results showed that DG and HQ at most of their combination ratios had synergic effect. Within the range of 1:5 - 5:1, all of the extracts of DG-HQ showed synergic effect, and among which, high-doses had better effects than low-doses. The highest value (-1) of the synergic effect was showed when DG was 10 - 40 g at the same time of HQ as 90 -180 g, and DG was 50 - 100 g at the same time of HQ as 20 - 100 g. DG-HQ at all combination dosages within Chinese Pharmacopeia (DG: 6 - 12 g, HQ: 9 - 30 g) had certain synergic effect, and Danggui Buxue Decoction (DG: 6 g, HQ: 30 g) also was at this range. The results provided scientific basis to the clinical application of DG and HQ. And the response surface method was firstly applied to quantitatively evaluate the bio-activity change of herb combination, which provided a novel way for modern basic research on the interaction of herbs.

Objective:To investigate the mechanism of microRNA-4298 (miR-4298) targeting inhibition of peptidylarginine deiminase 4 (PADI4) expression in U251 cells apoptosis induced by histone deacetylase inhibitor trichostatin A (TSA).Methods:The experiment was divided into TSA group (0.2 μmol/L TSA) and contral group. CCK8 method was used to detect the effect of TSA on the proliferation of U251 cells. Flow cytometry was used to detect the apoptosis level of U251 cells after drug action. Reverse transcription PCR and Western blotting experiments were used to determine the changes of PADI4 gene and protein expression after miR-4298 interference. Luciferase assay was used to determine the effects of miR-4298 on targeted binding and luciferase activity in the 3′UTR region of PADI4. U251 cells were transfected with PADI4 to observe the rescue effect of miR-4298 on apoptosis. Each experiment was divided into 3 groups, NC group, miR-4298 mimic group, TSA+ miR-4298 mimic group and NC group, miR-4298 inhibitor group, TSA+ miR-4298 inhibitor group.Results:U251 cells were treated with 0.2 μmol/L TSA for 4 days, the Absorbancy ( A) values of the control group was 1.168±0.148, which was higher than those of the TSA group (0.737±0.007), with statistically significant difference ( t=4.948, P=0.008). Compared with the control group, after treatment with 0.2 μmol/L TSA for 48 h, U251 cells showed obvious apoptosis (27.62%±3.49% vs. 4.99%±0.13%, t=11.190, P<0.001). Compared with those before the drug treatment, the PADI4 gene expression (0.386±0.020 vs. 0.903±0.021) and protein expression (0.276±0.041 vs. 0.777±0.031) after TSA treatment for 48 h both were decreased, with statistically significant differences ( t=30.400, P<0.001; t=16.770, P<0.001). The expression of miR-4298 during the process of TSA-induced U251 cell apoptosis increased (2.573±0.289 vs. 1.003±0.136; t=8.487, P=0.001). There was a negative correlation between the miR-4298 expression and PADI4 expression ( r=-0.877, P=0.002). Luciferase experiment confirmed that miR-4298 has targeted binding and luciferase inhibitory effects on the 3′UTR region of wild-type PADI4. Compared with NC group (0.920±0.026), the relative expression levels of PADI4 gene of miR-4298 mimic group (0.413±0.049) and TSA+ miR-4298 mimic group (0.213±0.035) were decreased, with statistically significant differences (all P<0.001). The PADI4 protein expression level change was consistent with the gene change trend. Compared with NC group (3.78%±0.68%), the apoptosis levels of miR-4298 mimic group (7.96%±1.10%) and TSA+ miR-4298 mimic group (13.74%±1.26%) were increased, with statistically significant differences ( P=0.005; P<0.001). Compared with NC group (0.183±0.025), the PADI4 gene expression of miR-4298 inhibitor group (0.483±0.032) and TSA+ miR-4298 inhibitor group (0.386±0.025) were increased, with statistically significant differences ( P<0.001; P=0.015). The PADI4 protein expression level change was consistent with the gene change trend. Compared with NC group (4.96%±0.59%), the apoptosis levels of miR-4298 inhibitor group (23.83%±2.20%) and TSA+ miR-4298 inhibitor group (9.55%±1.49%) were increased, with statistically significant differences (all P<0.001). In the rescue experiment, the expression level of PADI4 in miR-4298 mimic group was significantly increased ( P<0.001), while the expression level of PADI4 in miR-4298 mimic+ PADI4 group was relatively reversed ( P=0.002). Conclusion:miR-4298 can participate in the process of U251 cell apoptosis by targeting the expression of PADI4 gene, and miR-4298 may be the target of targeted intervention therapy for glioma.

Objective To investigate the food allergy situation of children in Changping District of Beijing,and to explore the influence of allergic family history,gender and mode of delivery on food allergy in children,the distribution of food allergy in different age groups,the types of food allergy that are easy to cause in this area,and the comorbidities of food allergy.Methods A total of 515 children aged 0 to 14 years who were admitted to the general pediatric outpatient and emergency department and inpatient of Beijing Changping Hospital from April to November 2023.Using immunoblotting to detect specific immunoglobulin E in the serum of pediatric patients,and using SPSS 26.0 statistical software to perform binomial tests on gender and delivery mode non parameters;Using custom Excel functions to statistically analyze the family history of allergies,the number of people in different age groups,and the frequency of allergic foods in each group;Use a self-made mini program to statistically analyze the combination of comorbid allergic diseases.Results ① The number of male children with food allergies(306 cases)was higher than that of female children(209 cases),and the difference was statistically significant(P＜0.05).(2)There were 109 cases of pediatric patients with parents who had no history of allergic diseases,accounting for 21.17％of the total cases;There were a total of 406 cases where at least one parent had a family history of allergies,accounting for 78.83％of the total cases.Among them,228 cases(44.27％)had one parent with a history of allergies,and 178 cases(34.56％)had both parents with a history of allergies.③ Among 515 children with positive food allergens,there were 10 cases(1.94％)in infancy,37 cases(7.19％)in early childhood,235 cases(45.63％)in preschool,192 cases(37.28％)in school age,and 41 cases(7.96％)in adolescence.The highest positivity rate for food allergens is in milk(444 times),followed by egg white(70 times),cashew nuts(57 times),crab(37 times),beef(26 times),mango(24 times),shrimp(21 times),pineapple(6 times),and shellfish(1 time).Milk protein is the most common allergen in all age groups.(4)Among 515 children with food allergies,399 cases were single food allergies,accounting for 77.47％;116 cases of multiple food allergies(2 or more types of food allergies),accounting for 22.53％.⑤ The most common comorbidity of food allergies is food allergy related gastrointestinal diseases combined with allergic rhinitis,with a total of 267 cases;Secondly,there were 192 cases of allergic rhinitis combined with chronic cough,and 124 cases of food allergy related gastrointestinal diseases combined with chronic cough.Conclusion Milk is the main allergen of food allergy in people of age 14 and under,and gastrointestinal symptoms are the most common in children with food allergy.