Objective To investigate the structure-activity relationship for 21 anthocyanins in inhibiting oxidized injury of endothelial cells,and explore the structural characteristics of anthocyanins closely related to their effects. Methods Endothelial cells were treated by ox-LDL at different concentrations of 50,100,150 or 200 ?g/ml,and MTT assay was used to determine IC50. After pre-incubated for 2 h with different concentrations ( 50,100 or 200 ?mol/L) of anthocyanins and then treated with 100 ?g/ml ox-LDL for another 24 h in endothelial cells,MTT assay was used to detect the cellular viability. After pre-treated for 2 h with different anthocyanins with 100 ?mol/L and treated with ox-LDL for another 24 h,MDA and NO level in the culture media were both measured according to the methods of assay kits. Structure-activity relationship was analyzed according to the respective cellular viability,MDA and NO level. Results Cellular viability was significantly inhibited by ox-LDL in a dose-dependent manner,and the IC50 was 100 ?g/ml. A significant correlation was observed among the effect of anthocyanins on cell viability,MDA production and NO release. The inhibitory effects of anthocyanins in ox-LDL-injured endothelial cells were positively related to the total number of hydroxyl groups and hydroxyl substitutions in B ring. 3′,4′-ortho-dihydroxyl substitution on B-ring and a 3-hydroxyl group on C-ring significantly enhanced the inhibitory effect of anthocyanins,yet methoxylation or glycosylation significantly decreased the effect. 6-hydroxylation substitution might attenuate the inhibitory effect of anthocyanins,while substitution at C5 or C5′ showed no significant influence on the effect of anthocyanins. Anthocyanin with monosaccharose substitution was much stronger than that with disaccharose substitution,while there was no significant difference between anthocyanins with glucoside and that with galacotoside substitution. Delphinidin and delphinidin-3-glucoside were respectively the most effectual anthocyanidin or anthocyanin. Conclusion 3′,4′-ortho-dihydroxyl substitution on B-ring and a 3-hydroxyl group on C-ring are the main structural requirements for anthocyanins in suppressing ox-LDL-induced injury in endothelial cells.

Developmental Dysplasia of the Hip ; Hip Dislocation, Congenital/therapy* ; Humans ; Infant ; Magnetic Resonance Imaging

Objective To investigate the relationship between single nucleotide polymorphisms(SNPs)in the eNOS gene and genetic susceptibility to systemic lupus erythematosus(SLE)in Hainan.Methods Blood samples were collected from SLE patients(SLE group,n=214)and healthy controls(control group,n=214)from January 2020 to December 2022 at the First Affiliated Hospital of Hainan Medical College and Hainan Provincial People's Hospital.The bases of eNOS gene rs3918188,rs1799983 and rs1007311 loci in each group were detected by SNaPshot sequencing technology.Logistic regression was used to analyze the correlation between genotypes,alleles and gene models(dominant model,recessive model,and overdominant model)of the above 3 target loci of the eNOS gene and genetic susceptibility to SLE.Haplotype analysis was conducted using HaploView 4.2 software to investigate the relationship between haploid and genetic susceptibility to SLE at each site.Results The results of logistic regression analysis revealed that the CC genotype and the C allele at rs3918188 locus were risk factors for genetic susceptibility to SLE(CC vs.AA:OR=2.449,P<0.05;C vs.A:OR=2.133,P<0.001).In recessive model at rs3918188 locus,CC genotype carriers had an increased risk of SLE development compared with AA+AC genotype carriers(OR=2.774,P<0.001).In contrast,in overdominant model at this locus,AC genotype carriers had a decreased risk of SLE occurrence compared with AA+CC genotype carriers(OR=0.385,P<0.001).In addition,polymorphisms of rs1799983 and rs1007311 were not associated with susceptibility to SLE in genotype,allele type and the 3 genetic models(P>0.05).Haplotype analysis revealed a strong linkage disequilibrium between the rs1007311 and rs1799983 loci of the eNOS gene,but no significant correlation was found between haplotype and genetic susceptibility to SLE(P>0.05).Conclusion The CC genotype and C allele at rs3918188 locus of eNOS gene may be risk factors for SLE in Hainan,while the risk of SLE occurrence is reduced in carriers of AC genotype under the overdominant model.

OBJECTIVE: To observe the effects of acupuncture combined with phacoemulsification for cataract with glaucoma. METHODS: Sixty-eight patients with cataract and angle-closure glaucoma were randomly divided into an observation group and a control group, 34 cases in each one. The patients in the control group were treated with phacoemulsification. Based on the treatment in the control group, the patients in the observation group were treated with and acupuncture at acupoints around eyes; the treatment was given once every other day, and 15-day treatment constituted a course, and 2 courses (60 days) were given. The intraocular pressure (IOP), visual acuity, pain severity, aqueous outflow (F value) and clinical symptoms were observed in the two groups before treatment and after 1 d, 7 d, 30 d and 60 d of treatment. RESULTS: The IOP after 1 d, 7 d, 30 d and 60 d of treatment in the observation group was lower than that in the control group (all <0.05), while the visual acuity after 7 d, 30 d and 60 d of treatment in the observation group was higher than that in the control group (all <0.05). Compared before treatment, the IOP was significantly decreased after 1 d, 7 d, 30 d and 60 d of treatment in the two groups, and the visual acuity was significantly increased after 7 d, 30 d and 60 d of treatment in the two groups (all <0.05). The pain scores and F value after 7 d, 30 d and 60 d of treatment in the observation group were lower than those in the control group (all <0.05). Compared before treatment, the pain scores and F value after 7 d, 30 d and 60 d of treatment were significantly reduced in the two groups (all <0.05). After 60 d of treatment, the number of patients who presented general clinical symptoms in the observation group was significantly lower than that in the control group (<0.05). CONCLUSION Based on phacoemulsification, acupuncture and could significantly reduce the IOP and F value, relieve pain and improve visual acuity in patients with cataract and glaucoma.
Acupuncture Therapy ; Cataract ; complications ; therapy ; Glaucoma ; complications ; therapy ; Humans ; Phacoemulsification ; Treatment Outcome

Objective To explore the effect of chenodeoxycholic acid(CDCA)on the expression of glucagon-like peptide-1(GLP-1)in the intestine of mice induced by high-fat diet(HFD)through farnesoid X receptor(FXR),and investigate the related mechanism.Methods Forty C57BL/6 mice were divided into control group,HFD group,HFD+CDCA group,HFD+Z-Gug(FXR antagonist)group,and HFD+CDCA+Z-Gug group,with 8 animals in each group.During intervention for 8 weeks,body weight and 24-hour food intake were measured every week.At the 8th week,oral glucose tolerance test(OGTT)and intraperitoneal glucose tolerance test(IPGTT)were conducted.After the mice were sacrificed,the serum levels of GLu,TG,CHO,LDL-C and HDL-C were detected;the expression levels of GLP-1 and FXR in intestinal tissues were detected by immunofluorescence assay;and the mRNA levels of TNF-α,IL-6,IL-1β,Gcg and FXR were detected by RT-qPCR;the serum level of GLP-1 was detected by ELISA,and the proportion of intraepithelial lymphocytes(IELs)subsets and the expression of CD26/DPP4 were detected by flow cytometry.Results Compared with the control group,the HFD group had increased body weight,abnormal serum glucose and lipid metabolism,impaired oral glucose tolerance,and weakened secretion of gastrointestinal hormones(P＜0.05),enhanced FXR expression at mRNA and protein levels,declined Gcg mRNA level and GLP-1 secretion level(P＜0.05),increased mRNA levels of intestinal inflammatory factors TNF-α,IL-6 and IL-1β(P＜0.05),raised proportions of TCRαβ+IELs,TCRαβ+CD8αα+IELs,and TCRαβ+CD8αβ+IELs but reduced proportion of TCRγδ+IELs,and increased total CD26/DPP4 expression in IELs(P＜0.05).Compared with the HFD group,HFD+CDCA treatment resulted in significantly increased body weight,impaired oral glucose tolerance,decreased secretion of gastrointestinal hormones,increased FXR mRNA and protein expression,and decreased Gcg mRNA expression and GLP-1 secretion(P＜0.05);decreased proportions of TCRαβ+IELs,TCRαβ+CD8αα+IELs and TCRααβ+CD8αβ+IELs but increased proportion of TCRγδ+ cells in IELs,and increased expression of total CD26/DPP4 in IELs(P＜0.05),which were significantly improved after Z-Gug intervention(P＜0.05).Conclusion CDCA may inhibit the expression and secretion of GLP-1 in intestinal tissue by activating FXR,and reduce the secretion of GLP-1.At the same time,CDCA may inhibit the expression of related inflammatory factors,regulate the proportions of IELs subsets,up-regulate the expression level of CD26/DPP4,promote the degradation of GLP-1 and aggravate insulin resistance.

Aim To investigate the protective effect of Huoxue Jiedu recipe on autophagy injury of H9C2 cardiomyocytes induced by hypoxia/reoxygenation and its mechanism. Methods H9C2 cardiomyocytes were used to establish a hypoxia/reoxygenation injury model. The effective concentration was screened and the cell activity was detected by CCK8 assay. The apoptotic rate of myocardial cells was detected by flow cytometry. The expression of autophagy marker LC3 was observed by laser confocal microscopy. The mRNA levels of Beclin-1, LC3 and Bcl-2 were detected by real-time quantitative PCR. The expressions of Beclin-1, LC311/I, Cleaved caspase-3, β-catenin, p-p65, Bcl-2, p62, p-Akt, p-mTOR were detected by Western blot. Results Huoxue Jiedu recipe can enhance the growth activity of myocardial cells and reduce the apoptotic rate and autophagy level, and it can enhance the activation of PI3K/Akt/mTORCl pathway, decrease Beclin-1 and LC3 mRNA levels, while increase Bcl-2 mRNA levels. It also decreased the expression of Beclin-1, LC311/I, Cleaved caspase-3, β-catenin, p-p65, and increased the expression of p62, p-Akt, p-mTOR, and Bcl-2. Conclusions Huoxue Jiedu recipe can reduce the level of autophagy and apoptosis of myocardial cells by regulating the autophagy pathway of PI3K/Akt/mTORCl, thereby playing a protective role in hypoxia/reoxygenation H9C2 myocardial cells.

The purpose of this study was to investigate the effect of inhibition of calpain on retinal ganglion cell-5 (RGC-5) necroptosis following oxygen glucose deprivation (OGD). RGC-5 cells were cultured in Dulbecco's-modified essential medium and necroptosis was induced by 8-h OGD. PI staining and flow cytometry were performed to detect RGC-5 necrosis. The calpain expression was detected by Western blotting and immunofluorescence staining. The calpain activity was tested by activity detection kit. Flow cytometry was used to detect the effect of calpain on RGC-5 necroptosis following OGD with or without N-acetyl-leucyl-leucyl-norleucinal (ALLN) pre-treatment. Western blot was used to detect the protein level of truncated apoptosis inducing factor (tAIF) in RGC-5 cells following OGD. The results showed that there was an up-regulation of the calpain expression and activity following OGD. Upon adding ALLN, the calpain activity was inhibited and tAIF was reduced following OGD along with the decreased number of RGC-5 necroptosis. In conclusion, calpain was involved in OGD-induced RGC-5 necroptosis with the increased expression of its downstream molecule tAIF.
Animals ; Apoptosis Inducing Factor ; biosynthesis ; genetics ; Calpain ; biosynthesis ; genetics ; Gene Expression Regulation ; drug effects ; Glucose ; metabolism ; Humans ; Leupeptins ; administration & dosage ; Mice ; Oxygen ; metabolism ; Retinal Ganglion Cells ; metabolism ; pathology ; Retinal Necrosis Syndrome, Acute ; genetics ; pathology

Objective: To investigate the relationship between genetic polymorphisms of cytochrome P450 2C19 (CYP2C19) and the efficacy of Helicobacter pylori (Hp) eradication therapy in children. Methods: The retrospective cohort study was conducted on 125 children with gastroscopy and positive rapid urease test (RUT) from September 2016 to December 2018 who presented to the Children's Hospital of Zhejiang University School of Medicine due to gastrointestinal symptoms including nausea, vomiting, abdominal pain, bloating, acid reflux, heartburn, chest pain, vomiting blood and melena. Hp culture and drug susceptibility test were carried out with gastric antrum mucosa before treatment. All the patients completed 2 weeks of standardized Hp eradication therapy and had ¹³C urea breath test 1 month after that, which was used to evaluate the curative effect. The DNA of gastric mucosa after RUT was analyzed and CYP2C19 gene polymorphism was detected. Children were grouped according to metabolic type. Combined with the results of Hp culture and drug susceptibility, the relationship between CYP2C19 gene polymorphism and the efficacy of Hp eradicative treatment was analyzed in children. Chi square test was used for row and column variables, and Fisher exact test was used for comparison between groups. Results: One hundred and twenty five children were enrolled in the study, of whom 76 were males and 49 females. The genetic polymorphism of CYP2C19 in these children found poor metabolizer (PM) of 30.4% (38/125), intermediate metabolizer (IM) of 20.8% (26/125), normal metabolizer (NM) of 47.2% (59/125), rapid metabolizer (RM) of 1.6% (2/125), and ultrarapid metabolizer (UM) of 0. There were statistically significant in positive rate of Hp culture among these groups (χ²=124.00, P<0.001). In addition, the successful rates of Hp eradication in PM, IM, NM and RM genotypes were 84.2% (32/38), 53.8% (14/26), 67.8% (40/59), and 0, respectively, with significant differences (χ²=11.35, P=0.010); those in IM genotype was significantly lower than that in PM genotype (P=0.011). With the same standard triple Hp eradicative regimen, the successful rate of Hp eradication for IM type was 8/19, which was lower than that of PM (80.0%, 24/30) and NM type (77.3%, 34/44) (P=0.007 and 0.007, respectively). There was a significant difference in the efficacy of Hp eradication treatment among different genotypes (χ²=9.72, P=0.008). According to the clarithromycin susceptibility result, the successful rate of Hp eradication treatment for IM genotype was 4/15 in the sensitive group and 4/4 in the drug-resistant group (χ²=6.97, P=0.018). Conclusions: The genetic polymorphism of CYP2C19 in children is closely related to the efficacy of Hp eradication treatment. PM has a higher successful rate of eradication treatment than the other genotypes.
Female ; Male ; Humans ; Child ; Cytochrome P-450 CYP2C19/genetics* ; Helicobacter pylori ; Retrospective Studies ; Genotype ; Abdominal Pain

Objective: Epidemiological studies reveal that exposure to fine particulate matter (aerodynamic diameter ≤ 2.5 μm, PM Methods: EVs were isolated from the serum of healthy subjects, quantified Results: PM Conclusions EVs treatment promotes cell survival and attenuates PM
A549 Cells ; Air Pollutants/toxicity* ; Apoptosis/drug effects* ; Cell Survival/drug effects* ; Extracellular Vesicles ; Humans ; Male ; Middle Aged ; Particulate Matter/toxicity* ; Protective Agents/pharmacology* ; Proto-Oncogene Proteins c-akt/metabolism* ; Serum