Development of a colloidal gold based immunochromatographic strip for 8-OHdG detection / 生物工程学报

8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive and stable biomarker for evaluating DNA oxidative damage. A rapid and sensitive colloidal gold immunochromatographic strip was developed for 8-OHdG detection by a competitive method. The sample pad (glass cellulose film), bonding pad (glass cellulose film), nitrocellulose film and absorbent pad were pasted on the polyvinyl chloride (PVC) base plate to construct the test strip. Colloidal gold (AuNPs) was prepared by the reduction of chloroauric acid with sodium citrate. 8-OHdG antibody (Ab) was coated on the outer layer of AuNPs to form Ab@AuNPs as a probe. Bovine serum albumin (BSA) and 8-OHdG were conjugated with carbodiimide hydrochloride to prepare an artificial antigen, which was used as the coating antigen of detection line. Goat anti mouse polyclonal antibody IgG was used as the coating antibody of control line. The experimental parameters were optimized including the type of nitrocellulose membrane, the formula of loading solution, and the spraying amount of gold labeled antibody. The results showed that the appropriate nitrocellulose membrane was CN 95. The optimal loading solution included BSA (1%), Tween-20 (3%), sucrose (3%) and NaCl (0.9%). The optimal spraying amount of gold labeled antibody was 4 μL. 8-OHdG can be detected by the strip under visible light, and the level of 8-OHdG in urine can be preliminarily determined by comparing the color intensity of T line and C line. The 8-OHdG concentration in urine was further calculated by the gray value of T line and the threshold of detection was 2.55 μg/L. This colloidal gold immunochromatographic strip is simple, rapid and specific for detecting 8-OHdG in human urine to preliminarily evaluate the human status.

Correlation research between angiopoietin-like protein 4 and vascular endothelial growth factor in serum and lower limb arterial lesions in Type 2 diabetes mellitus / 中国综合临床

Objective:To investigate the relationship between angiopoietin-like protein 4(ANGPTL4) and vascular endothelial growth factor(VEGF) in serum and lower limb arterial lesions in Type 2 diabetes mellitus.Methods:Using the method of retrospective cohort study, 164 inpatients with type 2 diabetes admitted to the Department of Endocrinology, Dahua hospital, Xuhui District, Shanghai from October 2017 to December 2018 were selected as the type 2 diabetes group, and 69 healthy people in the same period were collected as the normal control group.According to the ankle brachial index<0.9, diabetic patients were divided into two groups: the group without lower extremity artery disease and the group with lower extremity artery disease.The levels of ANGPTL4 and VEGF in serum were measured by enzyme-linked immunosorbent assay.Results:(1)Compared with the normal control group: the concentration of ANGPTL4 in serum of lower extremity arterial disease group and no lower extremity arterial disease group((44.78±15.15), (47.46±10.43) μg/L) were lower than that of normal control group((52.87±12.74) μg/L), the difference was statistically significant (all P<0.05). The serum VEGF concentration in lower extremity arterial disease group and no lower extremity arterial disease group((340.98±76.18), (314.83±75.30) ng/L) were higher than that in normal control group((282.58±81.4) ng/L), the difference was statistically significant(all P<0.05). (2)Compared with no lower extremity arterial disease group: the concentration of ANGPTL4 in serum of lower extremity arterial disease group was lower than that of no lower extremity arterial disease group, but the difference was not statistically significant( P>0.05). The serum VEGF concentration in lower extremity arterial disease group was higher than that in no lower extremity arterial disease group, the difference was statistically significant( P<0.05). (3)Correlation analysis: Diabetes mellitus and diabetic course>10 years( OR 4.594, 95% CI2.540-8.311, P<0.001), fasting blood glucose>8 mmol/L( OR 2.353, 95% CI1.023-5.416, P=0.044), glycosylated hemoglobin>8%( OR 6.442, 95% CI 2.874-14.441, P<0.001) and the increase of VEGF( OR 3.745, 95% CI1.566-8.953, P=0.003) are risk factors for lower limb arterial disease in type 2 diabetes, while the increase of ANGPTL( OR 0.154, 95% CI0.064-0.372, P<0.001) can reduce the occurrence of lower limb arterial disease in type 2 diabetes. Conclusion:ANGPTL4 is a protective factor and VEGF is a risk factor of type 2 diabetes mellitus.Both of them influence each other and participate in its occurrence and development.

The effects of high-fat diet on liver damage and pyroptosis in autoimmune hepatitis / 中华传染病杂志

Objective To investigate the effect of high-fat diet(HFD)on liver damage caused by autoimmune hepatitis(AIH)in mice.Methods Fifty C57BL/6 male mice were divided randomly into four groups:standard chow(SC)group,HFD group,AIH + SC group and AIH+ HFD group.AIH model was built after feeding for one week and all mice were sacrificed after four weeks.Liver and spleen tissues and serum were collected. Liver histopathology was detected by HE staining. Serum alanine aminotransperase(ALT)and aspartate aminotransferase(AST)levels were measured.Western blot analysis and real-time polymerase chain reaction(PCR)analysis were used to test the expressions of NLR pyrin domain containing 3(NLRP3)and cysteinyl aspartate specific proteinase 1(Caspase-1).The concentrations of interleukin(IL)-6,IL-1β and tumor necrosis factor(TNF)-α were analyzed using enzyme linked immunosorbent assay technology.The amount of Th17 cells in spleen was analyzed by FACS.Means among groups were analyzed with one-way ANOVA.SNK-q analysis was used for groups with homogeneity of variance, while nonparametric test was used for groups with variance nonhomogeneity.Results Histologically,the H&E staining of liver tissue from HFD group showed adipose degeneration,and there was inflammation around vessel in AIH+SC group.Moreover,in AIH+HFD group,the inflammation was more serious with mildly adipose degeneration.Compared with SC group,serum levels of ALT and AST increased in HFD group and AIH +SC group,and greatest increase was observed in AIH+ HFD group.The differences were statistically significant(F=57.12 and 37.58, both P<0.05).The proportions of Th17 cells in SC group,HFD group,AIH+ SC group and AIH+HFD group were(2.98 ± 0.90)%,(6.89 ± 0.99)%,(6.47 ± 1.08)% and(9.96 ± 0.83)%, respectively.The differences among all groups were statistically significant(F=54.05,P<0.05).The concentrations of IL-1β,IL-6 and TNF-α in each group were as follows:SC:IL-1β[(7.62 ± 2.81)ng/L],IL-6 [(106.54 ± 53.08)ng/L],T NF-α[(107.26 ± 36.20)ng/L];HFD:IL-1β[(25.06 ± 7.09)ng/L],IL-6 [(220.11 ± 47.41)ng/L],TNF-α[(273.77 ± 33.62)ng/L];AIH+SC:IL-1β[(17.49 ± 5.68)ng/L],IL-6 [(260.73 ± 50.29)ng/L],TNF-α[(250.49 ± 81.63)ng/L];AIH+ HFD:IL-1β[(52.04 ± 10.22)ng/L], IL-6[(785.93 ± 70.91)ng/L],TNF-α[(913.97 ± 64.57)ng/L].The differences were statistically significant(F=44.66,242.15 and 233.49,respectively,all P<0.05).The expressions of NLRP3 and Caspase-1 were significantly increased in AIH+ HFD group than the other three groups(all P<0.05). Conclusions High-fat diet potentiates liver damage induced by autoimmune hepatitis,which might relate to the secretion of pro-inflammatory cytokines,the activation of Th17 cells and the NLRP3 inflammasome as well as pyroptosis.

"High detective rate of""metabolic inflammatory syndrome""in patients with type 2 diabetes" / 中华内分泌代谢杂志

Objective Metabolites produced by metabolic imbalance such as free fatty acids and lipopolysaccharides can result in a state of chronic low-grade inflammation, or metabolic inflammation, which plays an important role in the pathogenesis of atherosclerosis, type 2 diabetes, non-alcoholic fatty liver disease, and obesity. The above metabolic disorders are closely related with the metabolic inflammation, which always coexist. Therefore, we proposed the concept ofmetabolic inflammatory syndrome ( MIS). According to our study, patients with two or more metabolic disorders above could be diagnosed as MIS. The current research is aimed to investigate the prevalence of MIS and its components, and to compare the clinical values of MIS and metabolic syndrome ( MS) . Methods 2 001 in patients with type 2 diabetes from 6 hospitals in Shanghai were recruited in the current multi-center cross-sectional study. The diagnostic rates of MIS and MS and their components of both syndromes were compared. Results In the patients with type 2 diabetes, the detective rate of MIS was 96. 2%, which was higher than that of MS (71. 3%). Among 4 components of MIS, atherosclerosis showed the highest detective rate (75.6%). MIS[OR=2.252(95%CI1.026-4.942),P=0.043],atherosclerosis[OR=2.726(95% CI1.953-3. 804),P<0. 001], and MS[OR=1. 915 (95%CI 1. 444-2. 540),P<0. 01] were the risk factors of coronary heart disease. Conclusion With atherosclerosis, type 2 diabetes mellitus, non-alcoholic fatty liver disease, and obesity as its 4 components, MIS has a high detective rate in patients with metabolic disorders, and seems to be more sensitive than MS to distinguish inflammation-related metabolic diseases. The concept of MIS will promote the screening and prevention of atherosclerosis in its early stage.

Effect of social support and coping styles on the stress and mental health in relatives of patients with traumatic brain injury / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of social support and coping style on the stress and mental health in relatives of patients with traumatic brain injury.@*METHODS@#The stress, mental health, social support and coping style were investigated in 300 relatives of patients with traumatic brain injury by Relative Stress Scale, Symptom Checklist-90, Social Support Rating Scale and Simplified Coping Style Questionnaire in Changsha City.@*RESULTS@#The mental health problems in relatives of patients with traumatic brain injury were closely related to the levels of stress, the ways of coping and the social support. In addition to the direct eff ect of stress on mental health in relatives of patients, the ways of coping and social support functioned as a mediator in this regard. The value of mesomeric eff ect for coping styles and social support ranged from 23.6% to 43.0%, and social support had an advantage over the coping styles.@*CONCLUSION@#Social support and coping styles should be considered in psychological nursing program to prevent and adjust the mental distress in relatives of patients with traumatic brain injury, which is beneficial to the treatment and recovery for patients.

Design and application of real-time settlement system for medical insurance based on Outpatient One Card Solution / 医疗卫生装备

Objective To realize real-time medical insurance settlement system of Xi'an based on Outpatient One Card Solution to reduce the errors due to artificial accounting.Methods The system was developed with C# language and Oracle 10g, based on interface function dynamic library calling rule and HIS data structure requirements provided by Xi'an municipal medical insurance center.Results The system realized real-time settlement of the patient with medical insurance card.Conclusion The system simplifies medical insurance outpatient treatment flow and implements data communication.