Objective To observe the effects of quercetin liposome (LQ) on formation of advanced glycation end prod-ucts(AGEs)and receptor for advanced glycation end products (RAGE) in kidney of diabetic rats. Methods LQ was made by rotary evaporation, and the model of type 2 diabetic rats were established by being fed on high-sugar and high-fat diet combined with intraperitoneally injection of streptozotocin (STZ). Then type 2 diabetic rats were randomly divided into six groups:diabetic model group (group DM), low level of LQ group (group LQ-L ), medium level of LQ group (group LQ-M), high level of LQ group (group LQ-H), positive control group (group aminoguanidine, AG) and control group (group N). After 8 weeks of interventions, blood glucose, body weight, kidney hypertrophy index (KI), blood urea nitrogen (BUN) and serum creatinine (Scr) were measured in each group. ELISA was used to detect serum AGEs, and 24 h urine albumin. The pathologi-cal change of glomerular basement membranes was observed by PAS staining and the expressions of AGEs in kidney was as-sessed by immunohistochemical method. The transcription level of RAGE mRNA in kidney was determined by RT-PCR. Re-sults Compared with the group N, the level of blood glucose, KI, BUN, Scr, serum AGEs and 24 h urine albumin were in-creased significantly in group DM, while the level of body weight decreased. Also the volume of kidney glomerulus increased and glomerular basement membranes thickened, the transcription levels of AGEs and RAGE mRNA in kidney tissue in-creased in DM group (P<0.05). Compared with group DM, the level of blood glucose, KI, BUN, Scr, serum AGEs and 24 h urinary albumin decreased, while the level of body weight increased in all three LQ groups. Meantime, the change of patho-logical morphology of glomerular basement membranes reduced and the expressions of AGEs and RAGE mRNA in kidney tissue decreased in all three LQ groups. All changes in the medium LQ dose group were more obvious than those of other two LQ groups (P<0.05). Conclusion Similar to AG, LQ has effect on inhibiting the action of proteinum unenzymatic glycosyl-ation and on decreasing the production of AGEs in serum as well as the expression of RAGE mRNA in kidney. Therefore, LQ play important protective role in kidneys of diabetic rats.

Objective To investigate the effects of 17-methoxyl-7-hydroxy-benzene-furabcgakcone( MHBFC ) on isoproterenol-induced ventricular remodeling in mice. Methods Isoproterenol(ISO)was given subcutaneously(1 mg·kg-1, twice per day for 7 d)to induce ventricular remodeling in mice. Mice were divided into normal control group,model group, captopril group,MHBFC low and high-dose groups. 65 mg·kg-1 captopril was given by intragastric administration in captopril group,2. 5,5. 0 mg·kg-1 MHBFC were given by intravenous injection in MHBFC low and high-dose groups. At the end of the 7th day,the hearts of the mice were weighted,and myocardial hypertrophy index was expressed as heart weight/body weight, double kidneys weight/body weight and lung weight/body weight( HW/BW,KW/BW and LW/BW). Colorimetric method was used to determine the content of hydroxyproline( Hyp)in heart,the activity of superoxide dismutase( SOD)and the content of malondialdehyde( MDA)in serum. The histological changes were observed by HE and Masson’s staining,the changes of cross section area( CSA),collagen volume fraction,( CVF)and perivascular circumferential collagen area( PVCA)were determined. Results Compared with the model group,MHBFC potently inhibited cardiomyocyte hypertrophy,decreased the HW/BW, KW/BW and LW/BW,improved cardiac pathology changed,increased the of activity SOD,decreased the content of MDA in serum and the content of Hyp in heart tissue(P﹤0. 01 or P﹤0. 05),decreased the CVF and PVCA(P﹤0. 01). Conclusion MHBFC possesses protective effects against ISO-induced ventricular remodeling in mice,which may be related to its actions in reducing the oxidative stress and improving the antioxidant activity of the body.

Objective To study the effect of DNA damage induced by H2 O2 on the micronucleus frequency in lymphocytes. Methods Resting lymphocytes were treated with different levels of H2 O2 (10,50,100,1 000 μmol/L).1 000 μmol/L H2 O2 was added into mitogen-stimulated lymphocyte cultures at different time intervals.Then micronucleus rate was examined by the conven-tional culture method.Results There was no significant change of the micronucleus frequency in the experimental groups.Conclu-sion H2 O2 could induce lymphocyte DNA damage rapidly,but exerts no effect on the formation of micronuclei,which may be relat-ed to the type of DNA damage and rapid DNA repair.

Objective To establish a method for the isolation and identification of platelets.Methods 10 healthy volunteers were selected to collect the EDTA anticoagulant venous blood of 3 tubes,each tube was 2 ml,which was divided into the whole blood cell tube,platelet rich plasma (control group),and stepped centrifugal platelet extract (experiment group).Platelet was isolated by simple centrifugation method(PRP) and stepped centrifugal method.The two groups were full blood count and analyzed by microscopic morphology and platelet activity test.Leukocyte specific HGB gene and platelet mitochondrial ND1 gene content was analyzed by real time PCR.Results Platelets were extracted and detected in control group and experimental group.Platelets were found and white blood cells and red blood cells were not remained in experimental group.Platelets and sporadic white blood cells were found in control group.The platelet pick up rate of experiment group was significantly higher than control group,the difference was statistically significant.Experimental gene content HGB of experiment group was significantly lower than control group,the difference was statistically significant (t=-3.281,-2.865,P＜0.05).ND1 gene content of experiment group higher than the control group,the difference was not statistically significant.There was no significant difference for platelet activity test between experimental group and control group (t=-0.046,-0.799,P＞ 0.05).Conclusion A isolation and identification method of stepped centrifugal platelet was established.The method can be used for the study of platelet gene and the functional analysis of platelets.

Objective: To investigate the detection of a human leukocyte antigen-B (HLA-B) allele HLA-B*13:01 by dual allele-specific real-time polymerase chain reaction (PCR) in patients with trichlorethylene-induced dermatitis. Methods: A total of 20 patients with trichlorethylene-induced dermatitis who were admitted and treated from January 2014 to October 2016 were enrolled as case group, and 20 persons who underwent physical examination from January to October, 2016 were enrolled as control group. Peripheral cubital venous blood samples were collected from all subjects, and dual allele-specific real-time PCR was used to detect the HLA-B*13:01 gene. The two groups were compared in terms of the proportion of subjects carrying HLA-B*13:01 gene. Results: There were no significant differences between the case group and the control group in median age (25.0 years vs 27.0 years, Z=0.30, P>0.05) and the proportion of male subjects (60.0% vs 70.0%, χ²=0.44, P>0.05) . The mean time of exposure to trichloroethylene was 30.8 days in the case group, while the subjects in the control group were not exposed to trichloroethylene. The case group had a significantly higher frequency of HLA-B*13:01 gene than the control group (80.0% vs 20.0%, χ²=14.40, P<0.01) with an odds ratio of 16.00. Conclusion Dual allele-specific real-time PCR can be used for detection of the HLA-B*13:01 gene in patients with trichlorethylene-induced dermatitis.

OBJECTIVETo investigate the cytotoxicity of indium chloride (InCl₃) and its effects on micro-nucleus formation in primary human lymphocytes cultured in vitro.

METHODSThe CCK-8 assay was used to evaluate the cytotoxicity of 24 h exposure to different concentrations of InCl₃(4, 40, 80, 200, 500, and 1 000 µmol/L) in lymphocytes cultured in vitro. The cytokinesis-block method was used to determine the micronucleus level in lymphocytes exposed to different concentrations of InCl₃and the effects of anti-oxidant vitamin C on micronucleus frequency.

RESULTSLymphocytes exposed to InCl₃of no less than 500 µmol/L had significantly lower survival rates than those in the control group (P < 0.05). Lymphocytes exposed to 80 µmol/L InCl₃had a significantly higher micronucleus frequency than those in the control group (P < 0.05). However, there was no further increase in micronucleus frequency of lymphocytes exposed to 200 µmol/L InCl₃. Lymphocytes cultured in whole blood and exposed to 500 or 1000 µmol/L InCl₃had a significantly increased micronucleus frequency than those in the control group (P < 0.001). The increase in micronucleus frequency of lymphocytes induced by indium could be partially antagonized by 20 or 100 µmol/L vitamin C.

CONCLUSIONInCl₃can induce an increase in micronucleus frequency of primary human lymphocytes cultured in vitro, which might be associated with DNA damage induced by oxidative stress.

Cell Nucleus ; metabolism ; Cytokinesis ; DNA Damage ; Humans ; In Vitro Techniques ; Indium ; toxicity ; Lymphocytes ; drug effects ; Oxidative Stress

OBJECTIVETo study the effects of indium exposure on the relative content of mitochondrial ND1 gene in lymphocytes.

METHODSVenous blood was obtained from 14 healthy workers and anticoagulated with heparin. Blood lymphocytes were separated and divided into three tube cultures. For two tubes in the exposed group, indium chloride was added to final concentrations of 0.2 mmol/L and 0.8 mmol/L, respectively. For one tube in the control group, an equal volume of normal saline solution was added. After incubation for 72 h, the relative content of mitochondrial gene in each group was determined using quantitative real-time PCR.

RESULTSLymphocytes exposed to 0.8 mmol/L indium chloride had a significantly higher relative content of mitochondrial gene than those exposed to 0.2 mmol/L indium chloride and those in the control group (P < 0.05, P < 0.05).

CONCLUSIONLymphocytes exposed to a high concentration of indium and its compounds have an elevated relative content of mitochondrial ND1 gene, indicating increased oxidative DNA damage induced by exposure to a high concentration of indium and its compounds.

DNA Damage ; drug effects ; DNA, Mitochondrial ; genetics ; Humans ; Indium ; toxicity ; Lymphocytes ; drug effects ; NADH Dehydrogenase ; genetics ; Occupational Exposure

OBJECTIVE: To explore the correlation between human leukocyte antigen( HLA)-B~* 13 : 01 allele and liver dysfunction in patients with occupational medicamentosa-like dermatitis due to trichloroethylene( OMDT). METHODS: Twenty patients with OMDT were chosen as study subjects by using a convenient sampling method. The sequence-based genotyping method was used for detecting HLA-B~* 13 : 01 allele in the DNA samples from peripheral blood of all study subjects. The serum levels of total protein,albumin,total bilirubin,direct bilirubin and alanine aminotransferase,aspartate aminotransferase and alkaline phosphatase activities in patients were examined. The correlation between the number of HLA-B~* 13 : 01 alleles and the liver function indices was also analyzed. RESULTS: There were 16 patients carrying HLA-B~* 13: 01 allele. The serum total protein in the HLA-B~* 13: 01 carriers was higher than that of non-carriers( P < 0. 05). The serum total protein was positively correlated with the number of patients carrying HLA-B~* 13: 01 alleles( P < 0. 05). CONCLUSION: The degree of liver function damage in OMDT patients may be related to carrying the HLA-B~* 13: 01 allele.