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Objective To investigate the effect of aluminum on hair loss induc ed by fluoride in fluorosis mice.Methods Sixty male C57BL mice were divided into four groups according to body mass:control group,fluoride (F) group (F-100 mg/L),aluminum(Al) group(Al3+ 270 mg/L) and F + Al group(F-100 mg/L + Al3+270 mg/L).Mice were killed 1 month and 3 months after the experiment,respectively.Bone F content was detected by ion-selective electrode method.The level of bone Al was measured through inductively coupled plasma emission spectrum.Dental fluorosis and hair loss of mice were evaluated by visual method.Results One month after the experiment,no dental fluorosis and hair loss was found in all four groups.The content of bone F was the highest in F group [(2401.649 + 86.835) mg/kg],and the lowest in A1 group [(427.006 + 11.878) mg/kg].The levels of bone F in F + Al group and control group were (1210.332 + 19.531)mg/kg and (538.001 + 33.337)mg/kg,respectively.The difference was statistically significant between any two groups (all P ＜ 0.05).Three month after the experiment,all mice of F treatment group had dental fluorosis and hair loss(10/10).Alopecia areas were found in the neck and back regions only.There was no hair loss in control group,Al group and F + Al group.No dental fluorosis was found in both control and Al groups.Only 2 mice were found with dental fluorosis in F + Al group.The levels of bone F in F group,F + Al group,control group and Al group were (4098.645 + 58.842),(1888.165 ± 12.187),(876.258 + 14.462) and (662.385 ± 8.966) mg/kg,respectively.The difference was statistically significant between any two groups (all P ＜ 0.05).Conclusions The hair loss is found in fluorosis mice.Hair loss of mice is closely associated with the level of F exposure.Al can prevent the occurrence of hair loss induced by F in mice through reducing the accumulation of F.
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Objective To establish a stable primary culture of rat cerebellar granule neurons in vitro for further study the toxic effects of chronic arsenic exposure on cerebellar cells.Methods Cerebellar cortices were taken from brain of Wistar rat 5-7 day old after born under stereoscopic microscope.Single cell suspension was acquired after digestion and washing with trypsin (0.25％) and DNase Ⅰ solution,respectively.Granule cells were purified from other cells by differential velocity adherence method for two times.Rat cerebellar granule neurons were seeded in culture plate pre-coated with poly-L-lysine.Neurons growth,development and synaptic connections were observed daily.The neurons were identified by neuron specific enolase (NSE) immunofluorescence technique.Results The neurons were affixed to the culture plate in 24 hours,in reticular arrangement observed under contrast microscope.Granule cells gradually turned round from oval and outlines became clearer in 2-3 days.In 4-6 days,there were a wide range of synaptic connections among the neurons and a mature nerve cell network formed.A large quantity of cerebellar granule neurons was seen by NSE identification.Few bigger cells such as purkinjes cells and glial cell outlines were also seen in the same visual field.Conclusions This is a successful primary culture method for acquirement of rat cerebellar granule neurons.The method can provide experimental basis for future studies the toxic effects of chronic arsenic exposure on cerebellar cells.
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ObjectiveTo observe the morphological feature of myocardial changes in adult rat exposed to citreoviridin(CIT) with selenium and protein deficiency.MethodsAccording to 2 × 2 factorial design,48 healthy male Wistar rats aged 4-week were randomly divided into four groups:group Ⅰ (Se-Pro-CIT+,low selenium and low protein plus CIT group),group Ⅱ (Se-Pro-CIT-,low selenium and low protein without CIT group),group Ⅲ (Se+Pro+CIT+,adequate selenium and adequate protein plus CIT group),and group Ⅳ (Se+Pro+CIT-,adequate selenium and adequate protein without CIT group),12 rats in each group.After one week of normal adaptive feeding,all the rats were fed with selenium and protein deficiency feed for 2 months,and then the animals in group Ⅰ and group Ⅲwere fed with 8 mg·kg-1 ·d-1 of CIT for 2 more months,after that the CIT dose was increased to 10 mg·kg-1 ·d-1 for the final 2 weeks.At the end of the experiment,all the rats were sacrificed by femoral artery bleeding,and body and heart weight were measured.Heart weight index was calculated and histopathological changes were observed under light microscope.Results Heart weight indexes of the 4 groups(Se-Pro-CIT+,Se-Pro-CIT-,Se+Pro+CIT+ and Se+Pro+CIT- groups) were (3.65 ± 0.45) × 10-3,(3.05 ± 0.19) × 10-3,(3.83 ± 1.06) × 10-3 and (3.31 ± 0.52) × 10-3,respectively.The results of factorial analysis showed that the effects of CIT on heart weight index were statistically significant(F =8.524,P ＜ 0.05 ),the effects of Se + Pro were not statistically significant(F =1.347,P ＞ 0.05),and there were no interactions between the two factors (F =0,048,P ＞ 0.05).Morphologically,tissue fibrosis around branch coronary artery in group Ⅰ rats,plenty of cardiocyte pycnosis in group Ⅱ,and myocardial scattered necrotic foci in group Ⅲ were observed,accompanied by inflammatory cell infiltration in group Ⅲ,and normal myocardial structure in group Ⅳ rats.Conclusions Citreoviridin plays a major role in causing myocardial injury( degeneration and necrosis) and CIT combined with selenium and protein deficiency can aggravate the
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ObjectiveTo clarify the causative effect of citreoviridin toxins(CIT) as well as nutritional deficiency of selenium and protein on rat myocardial injury at biochemical level.Methods According to 2 × 2 factorial design,48 healthy male Wistar rats aged 4-week were randomly divided into four groups:exposed to CIT along with nutritional deficiency of selenium and protein,nutritional deficiency of selenium and protein,exposed to CIT and control groups.After the rats in each group began to be fed with selenium and protein deficiency and(or) adequate fodder for 3 months,8 mg/kg body weight of CIT was fed daily to the rats in the two CIT toxin groups for two months.After that the CIT dose was raised up to 10 mg/kg body weight each day within the final 2 weeks.At the experimental endpoint,all the rats were sacrificed by femoral artery bleeding after ether anesthesia,and serum and heart specimens were collected for biochemical analysis by detecting serum Tn-Ⅰ and albumin,serum activities of CK and GSH-Px,myocardial SOD and T-AOC.ResultsThe interactions between Se & protein and CIT in rat final body weight,serum albumin,and Tn-Ⅰ was observed(F=8.186,6.160,19.183,all P＜ 0.05),whereas interactions in rat body weight of 12 weeks,serum GSH-Px,CK as well as myocardial SOD,T-AOC activity were not found (F=1.633,1.987,0.075,0.474,1.145,all P ＞ 0.05).Under the nutritional deficiency of selenium and protein,serum albumin and Tn-Ⅰ level in the groups with CIT toxin[ (42.88 ± 1.19) g/L,(668.6 ± 55.8) ng/L,respectively]were lower than that of the group without CIT toxin[ (47.59 ± 1.05)g/L,(989.3 ± 49.2)ng/L,respectively,all P ＜0.05].The main effects of selenium and protein on rat body weight of 12 weeks,serum GSH-Px,myocardial SOD and T-AOC were statistically significant between different groups (F =96.860,58.086,4.475,25.485,all P ＜ 0.05).Rat body weights of 12 weeks in the two groups of nutritional deficiency of selenium and protein[ (186.33 ± 7.89),(197.83 ± 7.89)g] were all lower than others[ (274.08 ± 7.89),(265.42 ± 7.89)g,all P ＜ 0.05]; serum GSH-Pxs in the two groups of nutritional deficiency of selenium and protein[ (317.5 ± 102.6),(296.9 ± 90.5)U/L] were all lower than others[ (926.1 ± 110.9),( 1181.7 ± 85.9)U/L,all P ＜ 0.05] ; myocardial SODs in the two groups of nutritional deficiency of selenium and protein [ (65.22 ± 5.91 ) × 106, (62.68 ± 5.61 ) × 106 U/kg] were all lower than others [(74.07 ± 7.24) × 106, (80.07 ± 5.91) × 106 U/kg,all P＜ 0.05]; myocardial T-AOCs in the two groups of nutritional deficiency of selenium and protein[ (1.138 ± 0.086) × 106,(0.806 ± 0.081 ) × 106 U/kg] were all lower than others[(1.688 ± 0.105) × 106,(1.163 ± 0.086) × 106 U/kg,all P ＜ 0.05].Conclusions Animal model with selenium and protein deficiency is successfully established.However,the results of biochemical index tested in the experimental rats show no regularity effect,which needs to be checked again in the future study.
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Objective In this study,we investigated the relationship between oxidative stress,selenoproteins level and onset of Keshan disease (KD) through detecting the expression of 8-hydroxy-2-deoxy guanosine (8-OH-dG),thioredoxin reductase 1 (TrxR1) and glutathione peroxidase 1 (GPx1) in myocardial tissue.Methods Myocardium samples of autopsy patients including 8 cases of KD (KD group included 4 acute KD and 4 chronic KD) and 9 cases of non-KD (control group) were immunohistochemically stained for 8-OH-dG,TrxR1 and GPx1.The staining intensities subsequently quantified by using Olympus Image-Pro Plus 6.0 software.Results The positive rate of 8-OH-dG expression in myocardial nuclei was higher in the case group[(68.6 ± 20.4)％] than that of the control group[(2.4 ± 1.5)％,t =8.515,P ＜ 0.05].In addition,the positive rate of 8-OH-dG expression in acute KD[(91.7 ± 3.7)％] was significantly higher than that of chronic KD[(53.2 ± 7.9)％,t =6.409,P＜0.05].The distribution of TrxR1 and GPx1 was not associated with the distribution of myocardial damage.The expression of these two selenoproteins in KD group (401340 ± 59865,497590 ± 197082) were both lower than that of control group(2790300 ± 379298,1348400 ±615840; t =-28.493,-6.016,respectively,all P＜0.01).Conclusions Oxidative damage is detected in myocardium tissue of KD,and 8-OH-dG expression is associated with the degree of myocardial damage in KD.Selenoproteins,TrxR1 and GPx1,may be closely related to the pathogenesis of KD.
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ObjectiveTo explore the effect of selenium,protein and vitamin E deficiency on mRNA expression of rat cardiac selenoprotein,and their relation with myocardial injury.MethodsForty male Wistar rats were randomly divided into 4 groups:low selenium low protein low vitamin E group(group A),low selenium low protein adequate vitamin E group(group B),adequate selenium adequate protein low vitamin E group(group C),and adequate selenium adequate protein adequate vitamin E group (group D),10 rats in each group.The activity of whole blood glùtathione peroxidase(GSH-Px ) was measured using dithiobis nitrobenzoic acid (DTNB) at the end of sixth month experiment.The levels of mRNA expression of glutathione peroxidase 1(Gpx1),phospholipid hydroperoxide glutathione peroxidase 4(Gpx4),thioredoxin reductase(TrxR),selenoprotein P(Se-P) and selenoprotein W(Se-W) were determined by real-time fluorescence quantitative PCR at the end of sixth month.Histopathological changes of myocardial injury were observed with light microscope.ResultsThe activity of GSH-Px was (44.6 ± 3.1 ),(45.5 ± 1.6),(86.6 ± 2.2),(85.6 ± 1.2)U/L,respectively,in the above four groups at the end of sixth month,and the difference was statistically significant(F =100.7,P ＜ 0.01 ) ; the activity of GSH-Px of groups C and D was higher than that of groups A,B(all P ＜ 0.05).mRNA expression of myocardial tissue of the four groups was as follows,Gpx1(0.099 ± 0.312,0.054 ± 0.007,0.386 ± 0.067,0.340 ± 0.085),Gpx4(1.005 ± 0.089,0.810 ± 0.229,0.895 ± 0.084,0.922 ± 0.399),and Se-W(0.188 ± 0.080,0.119 ± 0.069,0.574 ± 0.167,0.570 ± 0.383),and the difference was statistically significant(F =112.1,3.76,22.8,all P ＜ 0.05) ; the mRNA levels of Gpx1,Se-W of groups C,D were significantly higher than that of groups A,B(all P ＜ 0.05).The mRNA expression of Gpx4 of group A was higher than that of group C(P ＜ 0.05).The mRNA expression of TrxR(0.130 ± 0.037,0.127 ± 0.038,0.134 ± 0.021,0.120 ± 0.014) and Se-P(0.446 ± 0.155,0.413 ± 0.152,0.385 ± 0.041,0.408 ±0.208 ) was not statistically different among the four groups (F =0.91,1.75,all P ＞ 0.05).Pathological changes of myocardial tissue were mainly as foci of coagulative necrosis.The necrosis detection rate of the four groups was 8/10,4/9,2/10,and 1/10,respectively,and the difference was significant statistically(Fisher exact test,P =0.0067).ConclusionsLong-term selenium,protein and vitamin E deficiency will reduce body antioxidant capacity and lead to myocardial injury.The mRNA levels of Gpx1 and Se-W and selenium level are closely related.The mRNA levels of Gpx4,TrxR and Se-P remain relatively stable.
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Objective To study the effects of selenium(Se) and protein on cardiac morphology and expression of cellular glutathione peroxidase(GPX1 ) and mitochondrial thioredoxin reductase(TR2) in rat myocardial tissue.MethodsSixty healthy weaning male Wistar rats were randomly divided into four groups by two factors two levels factorial design(n =15).Drinking water was divided into two levels of Se-deficient(0 mg/L) and Se-adequate (0.25 mg/L); diet was divided into two levels of protein-deficeient (10％ protein and 0.008 mg/kg Se) and protein-adequate(20％ protein and 0.015 - 0.026 mg/kg Se).The rats were killed after feeding for one year.Pathological changes in myocardial tissues were observed under light microscope.The expression of GPX1 and TR2 in rat myocardial tissue was detected by immunohistochemistry and Western blotting.Results Compared between groups,the difference of the rate of myocardial necrosis in rats was statistically significant(x2 =11.04,P ＜ 0.05),in which Se-deficient protein-deficient group [66.7％ (8/12) ] was significantly higher than Se-adequate proteinadequate group [ 7.1％ ( 1 / 14),x2 - 11.06,P ＜ 0.05 ].GPX 1 positive rates in Se-deficient protein-deficient group,Se-adequate protein-deficient group,Se-deficient protein-adequate group and Se-adequate protein-adequate group were 0(0/12),81.8％(9/11 ),10.0％(1/10) and 100.0％(14/14),respectively,in rat myocardial tissue determined by immunohistochemistry.Of which,Se-adequate protein-deficient group and Se-adequate protein-adequate group were significantly higher than Se-deficient protein-deficient group and Se-deficient protein-adequate group(x2 =12.88,8.14 and 35.89,32.60,all P ＜ 0.05).The positive expression rates of TR2 in rats myocardial tissue of the four groups were 0(0/12),81.8％(9/11),0(0/10) and 100.0％(14/14),respectively.Of which,Se-adequate proteindeficient group and Se-adequate protein-adequate group were significantly higher than Se-deficient protein-deficient group and Se-deficient protein-adequate group (x2 =28.67,18.25 and 35.89,32.60,all P ＜ 0.05).The four groups'results of the overall mean of the relatively value of protein expression of GPX1 in cardiac tissue by Western blotting were 0.87 ± 0.13,1.18 ± 0.13,0.95 ± 0.13 and 1.74 ± 0.23,respectively.Through analysis of variance of factorial design,the effects of Se and protein on protein expression of GPX1 in the heart were statistically significant(F=124.93,43.16,all P＜ 0.05).And there was interaction between them(F=24.10,P＜ 0.05).The four groups'results of the overall mean of the relatively value of protein expression of TR2 in cardiac tissue by Western blotting were 0.63 ± 0.19,0.97 ± 0.24,0.55 ± 0.08 and 1.03 ± 0.31,respectively.Through analysis of variance of factorial design,the effect of Se on expression of TR2 in the heart was statistically significant(F =36.97,P ＜ 0.05).Conclusions Adequate Se and protein diet can increase the levels of GPX1 and TR2 in the heart compared to deficient Se and protein diet,can enhance anti-oxidizing ability,protect the myocardial endothelial cells,reduce degree of myocardial injury,and the combined effects of both are better.
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Objective To observe protective effects on rat serum cardiac enzymes and the antioxidant capacity of selenium and vitamin E.Methods According to body weight and 2 × 2 factorial design,eighty male Wistas rats were randomly divided into four groups:low selenium and low vitamin E group(feed containing 23.42％ of the low selenium yeast,excluding vitamin E),low selenium and adequate vitamin E group (feed containing 23.42％ of the low selenium yeast and vitamin E 160 mg/kg),adequate selenium and low vitamin E group(feed containing 46.84％ of the low selenium yeast and sodium seleni 0.25 mg/L in water,excluding vitamin E),adequate selenium and adequate vitamin E group(feed containing 46.84％ of the low selenium yeast,vitamin E 160 mg/kg and sodium selenite 0.25 mg/L in water),20 rats every group.Rats were feed with synthetic feed,and given intraperitoneal anesthesia after 26 weeks of feeding.Blood was collected to observe the impact of selenium and vitamin E on rat cardiac enzymes and myocardial antioxidant capacity and their interactions.Serum creatine kinase (CK) was measured using the continuous monitoring method,creatine kinase isozymes (CK-MB) and lactate dehydrogenase(LDH ) using the immune suppression method,the whole blood GSH-Px assay using the dithiobis nitrohenzoic acid(DTNB) method,serum superoxide dismutase(SOD) using the xanthine oxidase method,total antioxidant capacity (T-AOC) using the complex colorimetry method,the content of propylene glycol (MDA) using the thiobarbituric acid colorimetric method,and reactive oxygen species(ROS) using the colorimetric method.Results Group differences of serum CK,CK-MB,LDH,whole blood GSH-Px activity,serum T-AOC vitality,MDA and ROS content were statistically significant(F=9.797,17.041,48.399,3.744,224.900,49.384,5.045,all P＜ 0.05).Compared with the two low selenium groups and one adequate selenium group,the vitalities of CK,CK-MB,LDH and the contents of MDA[(1577.75 ± 451.87),(1239.15 ± 344.99),(884.25 ± 133.84)U/L,(5.688 ±1.169) × 103 nmol/L; (1474.21 ± 398.38),(1014.84 ± 215.40),(523.00 ± 98.05)U/L,(4.035 ± 0.487 ) × 103 nmol/L and (1180.10 ± 245.51),(948.75 ± 173.68),(676.70 ± 193.63)U/L,(3.406 ± 0.146) × 103 nmol/L]increased significantly in adequate selenium and adequate vitamin E group[( 1056.80 ± 250.98),(721.70 ±129.98),(404.65 ± 72.49)U/L,(3.010 ± 1.270) × 103 nmol/L,all P ＜ 0.05) ].The activity of GSH-Px was obviously increased in the two adequate selenium groups[ (96.611 ± 8.238) × 103,(103.024 ± 8.217) × 103 U/L,all P ＜ 0.05],compared with the two low selenium groups[ (60.356 ± 8.179) × 103,(63.117 ± 8.281) × 103 U/L].Selenium affected the activities of CK,CK-MB and LDH(F =27.09,31.58,29.66,all P＜ 0.01 ),and vitamin E affected the activities of CK-MB and LDH(F=18.9,11.2.all P＜ 0.01 ),but both selenium and vitamin E had no interactions on the activities of CK,CK-MB and LDH (F=0.02,0.001,2.22,all P＞0.05).Selenium affected the activity of GSH-Px and the content of MDA(F=6.74,95.68,all P＜ 0.05),vitamin E affected the activity of T-AOC,the contents of MDA and ROS(F=6.42,36.73,8.43,all P＜0.05),but selenium and vitamin E had interactions only on the content of MDA(F =13.82,P＜ 0.05).Conclusions Long-term selenium or vitamin E deficiency,can reduce the body's antioxidant capacity,leading to the occurrence of myocardial injury.Selenium and vitamin E can improve the body's oxidation capacity,playing a role in myocardial protection.
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Objective To observe the effects of recombinant human parathyroid hormone 1 to 34(referred to as hPTH) on the expression level of alkaline phosphatase(ALP) and bone gla protein(BCP) in human osteosarcoma cell line SaOS-2(referred to as SaOS-2 cells). Methods SaOS-2 cells were subcultured and treated with 1, 10 and 100 nmol/L hPTH for 12, 24 and 48 h. Total cellular RNA was extracted, cDNA was synthesized by reverse doses of hPTH, different duration of action, and their interaction on the expression level of ALP mRNA of SaOS-2 cells was significantly different(F = 29.32, 2.92, 7.64, all P < 0.05). The expression level of ALP mRNA(0.78 ± 0.43, 0.71 ± 0.05, 0.75 ± 0.19, 0.76 ± 0.14) of SaOS-2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h(1.01 ± 0.16, 1.37 ± 0.38, 1.49 ± 0.16, 2.52 ± 0.70, all P< 0.05) and 24 h (1.80 ± 0.47, 1.30 ± 0.36, 1.27 ± 0.17, 1.17 ± 0.11, all P< 0.05). The expression level of ALP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 hours was higher than that of the control(P < 0.05); the expression level of ALP mRNA of SaOS-2 cells after treatment with 1, 10 and 100 nmol/L hPTH for 24 h interaction on the expression level of BGP mRNA of SaOS-2 were significantly different (F = 8.26, 10.33, 5.51, all P< 0.05). The expression level of BGP mRNA(1.17 ± 0.28, 0.98 ± 0.08, 0.92 ± 0.17 and 0.84 ± 0.59) of SaOS2 cells after treatment with 0, 1, 10 and 100 nmol/L hPTH for 48 h was lower than those of treated for 12 h( 1.01 ± 0.14, 1.21 ± 0.18, 1.34 ± 0.30, 1.68 ± 0.62, all P< 0.05), and 24 h(1.71 ± 0.35, 1.41 ± 0.47, 1.28 ± 0.31 and 1.01 ± 0.18, all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 12 h was higher than that of those groups treated with 0 and 1 nmol/L hPTH(all P< 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 10 and 100 nmol/L hPTH for 24 h and 48 h was lower than those of the control(all P < 0.05). The expression level of BGP mRNA of SaOS-2 cells after treatment with 100 nmol/L hPTH for 24 hours was lower than that the group treated with 1 nmol/L hPTH(P < 0.05). Conclusions In vitro, hPTH significantly enhances osteogenic activities of human osteoblast in a short time, however, with prolonged stimulation time, osteogenic activity can show a downward trend.
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Objective To investigate the effect of recombinant human bone morphogenetic protein 2 (rhBMP-2) on the osteogenic activities of human osteosarcoma cell line SaOS-2. Methods SaOS-2 cells were exposed to rhBMP-2 for 12,24,48 h at 0(control) ,2,20,200 μg/L, respectively. The mRNA expression of alkaline phosphatase(ALP) and bone gla(BCP) were detected by real time polymerase chain reaction. Results The mRNA expression of ALP and BGP of SaOS-2 cells increased gradually with rhBMP-2. The mRNA expression of ALP of the 20 μg/L group exposed for 48 h(1.60 ± 0.64), and the 200 μg/L group exposed for 12,48 h(1.70 ± 0.41, 1.80±0.19) were significantly higher than those of control (12 h: 0.80±0.25, 48 h: 0.74±0.21, allP<0.05). The mRNA expression of BGP of the 2 μg/L group exposed for 24 h(1.67 ± 0.33), the 20 μg/L group exposed for 12,24 h(2.42 ± 0.13,1.82 ± 0.14) and the 200 μg/L group exposed for 12,24 h(1.46 ± 0.11,1.24 ± 0.07) were significantly higher than those of control( 12 h: 1.01 ± 0.14, 24 h: 0.84 ± 0.12, all P< 0.05). Conclusions rhBMP-2 can promote the mRNA expression of ALP and BGP of SaOS-2 cells. They have a dose-response relationship, but represent a different dose-response effect.
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Objective To ohserve the rat myocardial damage induced by citreoviridin(CIT)in the status of combined selenium and protein deficiency.Methods According to 2×2 factorial design,forty 4-week-old healthy Wistar rats were randomly divided into four groups.i.e.combined selenium and protein adequate with no CIT and with some CIT groups(Se+Pro+CIT-.Se+Pro+CiT+),combined selenium and protein deficiency with no CIT and with some CIT groups(Se-Pro-CIT-,Se-Pro-CIT+).The numbers of male and female were fifty-fifty.Theserats were fed with combined selenium and protein adequate and combined selenium and protein deficiency fodder until the 16th week. Cardiac toxicity of CIT was evaluated by general state of health, heart weight index, myocardial pathological change, the levels of selenium and the activities of glutathion peroxidase (GSH-Px) and creatine kinase (CK) in serum, and the activity of superoxide dismutase(SOD) of myocardium. Results The interaction effects of combined selenium and protein deficiency and adequate CIT on body weight, serum levels of selenium and albumin, heart weight index, the activities of CK and GSH-Px in serum and SOD of myocardium were statistically not significant(F= 0.000, 1.210, 0.625, 0.981, 2.785, 0.074, 0.001, all P> 0.05). The main effects of combined selenium and protein on the levels of serum selenium and albumin, heart weight index and the activity of GSH-Px in serum were statistically significant(F = 507.698, 87.734, 4.201, 109.389, all P < 0.05). The main effects of CIT on body weight, the levels of serum selenium and albumin, heart weight index and the activity of CK in serum were statistically significant(F = 10.929, 4.371, 26.108, 24.844, 4.439, all P < 0.05). The mean levels of serum selenium of Se-Pro- groups [(70.4 ± 40.0), (87.7 ± 59.6 )μg/L] were lower than those of Se+Pro+ groups [(446.1 ± 74.8),(502.1 ± 39.2)μg/L, all P < 0.05]. The mean levels of serum albumin of Se-Pro- groups [(34.36 ± 1.28 ), (33.38 ±2.48)g/L] were lower than those of Se+Pro+ groups[(40.69 ± 1.30), (38.71 ± 2.15)g/L, all P < 0.05]. The mean levels of heart weight index of CIT+ groups[(4.14 ± 0.36) × 10-3, (4.39 ± 0.53) x 10-3] were higher than those of CIT-groups[(3.56 ± 0.26) x 10-3, (3.80 ± 0.28) x 10-3, all P < 0.05] respectively at the same levels of selenium and protein. The mean levels of CK in serum of Se-Pro-CIT+ group[(2.54 ± 0.56)kU/L] was lower than that of Se-Pro-CIT- group [(3.37 ± 0.67 )kU/L, P < 0.05]. The mean levels of activity of GSH-Px in serum of Se-Progroups[(408.1 ± 412.6), (510.5 ± 392.0)U/L] were lower than those of Se+Pro+ groups[(1667.8 ± 102.2),(1731.5 ± 144.4)U/L, all P < 0.05]. In Se+Pro+CIT+ group, there was part of intercalary disc of cardiac myocytes fragmented;the conjunctions between myoeytes were broken;in some region, cardiac myocytes became edematous,even dissolved. In Se-Pro-CIT- group, the change of cardiac myocytes membrane structures was not obvious;filament structure was disappeared around nucleus;deposition of mass floccule could be seen. In Se-Pro-CIT+ group,the structure of sarcomeres was not obvious;mitochondrial cristae was loosened;cavities in myocytes could be seen occasionally;there were lots of disseminated sareoplasmic reticulum extending. Conclusions .CIT is the main risk factor in inducing myocardial damage. The deficiency of combined selenium and protein can aggravate the damage,but its independent pathogenic effect is weak.
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Objective To explore the relationship between myocardial damage and antioxidant capacity in vivo of Keshan disease patients and to analyze the possible pathogenesis. Methods In the period from 2005 to 2006, 41 chronic and latent Keshan disease cases were chosen as the case group from such serious endemic areas as Yongjin Village, Xinfa Village of Fuyu County, Fuan Village of Shangzhi County, Xinghuo Village of Wudalianchi County, 61 healthy people from the same area as internal controls, 48 healthy people from Xianglansan Village of Wangkui County, an un-endemic area, as external control. Fasting peripheral venous blood was collected from all the people. And blood selenium, glutathione peroxidase(GSH-Px) and suporoxide dismutase.(T-SOD) activities, malondialdehyde (MDA) levels were examined. Results Blood selenium level of the patient group [(34.80±13.30) μg/L], GSH-Px[(104.10±34.19)U/L]and T-SOD[(92.16±17.98)×103 U/L]actives were significantly lower than the internal control group [(41.24±13.57)μg/L, (118.57±25.49)U/L, (104.82±13.56)×103 U/L]and the external control group [(48.33±16.51)μg/L, (155.00±24.01)U/L, (108.48±12.73)×103 U/L], respectively, with a statistically significant difference(all P < 0.05). MDA level of the patient group[(7.12± 1.37)μmol/L]was higher than that in the internal control group[(5.36±1.18)μmol/L]and the external control group[(5.22±0.83)μmol/L]with a statistically significant differences(both P < 0.05). The blood selenium level, GSH-Px activity of internal control group was resoectively lower than that in the external control group, the differences being statistically significant(beth P < 0.05). Conclusions Selenium deficiency may lead to reduced antioxidant capacity and enhanced oxidative damage in Keshan disease patients in vivo. There may be a certain relationship between oxidative stress disorder and myocardial damage of Keshan disease.
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Objective To investigate the myocardial damage in rats fed with corn from Keshan disease area added with bean puree. Methods Male Wistar rats were randomly divided into 3 groups according to their body weights, and fed with corn, corn from Keshan disease area added with bean puree, corn from non-endemic area. The GSH-Px activity of vena cardalis blood was examined in 1 and 3 months, rats were sacrificed after being fed for 6 months to examine the heart changes with HE stain. Results The three groups of GSH-Px activity were different in 1 and 3 months respectively(F=23.60,72.46, all P<0.01); GSH-Px activity was (181.58±22.15), (44.76±28.59)U/L in rats fed with corn, was (195.03±17.66), (30.38±3.35)U/L in those fed with corn added with bean puree from Keshan disease area, lower than the group fed with corn of non-endemic area[(340.90±95.42), (125.17±13.64)U/L, all P < 0.01]. But the difference of GSH-Px activity between simple corn group and corn adding bean puree groups of Keshan disease area was not obvious(P>0.05). Myocardial damage incidence of the three groups was 3/9,1/9,2/7. Difference among three groups did not have statistical significance(χ2=1.33, P> 0.05). Conclusions Only corn from Keshan disease area may induce myocardial damage pathology change. Adding bean puree into corn does not increase damage.
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Objective To explore the inhabitants' knowledge about Keshan disease(KD) in endemic area in Heilongjiang Province and analyze its influencing factors. Methods Yongjin Village, a national KD monitoring site in Fanrong Township of Fuyu County in Heilongjiang Province,was selected as an investigating spot. Specially designed questionnaire was applied in the investigation, which included three aspects, demographical information (gender, age, educational level, etc.), KD-related knowledge survey (KD prevention measures including a healthy diet supplementing selenium, preventing microbial infection, inducers such as climate change, fatigue, cold, mental shock, overeating) and KD controlling measures (take prescribed medicine, dietetic regulation and regular exercise, etc.) Results Seventy-senven point six percent(272/352) knew the disease by the name in the endemic area, while only 29.7% (81/272)and 30.8% (84/272)were aware the preventive measures and inducers , respectively. The awareness rates of KD preventive measures and inducers among the KD patients were 6/17 and 9/17 respectively. The influencing factors of the awareness rate of KD name were gender, vocation and the state of education (χ2 value were 9.838,9.878,12.462, all P < 0.05). However, factors influencing the awareness rates of preventive measures were gender and the state of education(χ2 value were 7.400,20.251, both P < 0.05). Conclusions The awareness rates of KD preventive measures and inducers axe low in the endemic area. The major factors influencing the awareness rate of KD are gender and the educational degree.
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Objective To explore the potential risk factors to Ymman endemic sudden cardiac death (YESCD) and provide evidence for prevention. Methods Twenty-two cases and 24 controls were randomly selected from YESCD areas and non-YESCD areas, respectively. Both cases and controls were interviewed with unified questionnaires. Univatiate X2 test and multivariate conditional logistic stepwise regression was conducted with SPSS 13.0. The optimal regression model was established and evaluated. Results The univariate X2 teat revealed that presence or absence of 5 potential factors might possibly be associated to YESCD: appropriate places for storing dining utensils, pens for livestock, consumption of mushrooms, exposure to any pesticides, and sudden climate changes before onset(X2=12.206,4.779,5.741,6.120,10.754, P<0.05). Multivariate conditional logistic stepwise regression demonstrated that consumption of mushrooms was a protective factor(OR=0.115, P<0.05) and sudden climate change was a risk factor(OR=36.592, P<0.01). Conclusions Sudden climate change might be a risk factor contributing to YESCD, and eating mushrooms before the prevalence seasons may provide some protection.
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Objective To observe the change in cardiac shape and heart function and evaluate the effect of self-treatment on patients with Keshan disease by echocardiography. Methods To check the 31 patients with Keshan disease before the self-treatment, and follow them up in the 3rd and 6th months after self-treatment by echocardiography. The left atrium diameter(LAd), left ventricular end-diastolic diameter(LVEDd), the thickness of interventricular septum in end-diastolic(IVSTd), the thickness of LV posterior wall in end-diastolic (LVPWTd), left ventricular mass(LVM), left ventricular mass index(LVMI), left ventricular ejection fraction(LVEF) and mitral valve flow E/A ratio(E/A) were measured. Results The LAd[(35.8±5.1)ram] and LVPWTd[(9.3±1.0)mm] obviously decreased in the 3rd month after serf-treatment compared with prior self-treatment [ (37.0±5.0), (9.9± 1.2)mm](P<0.05). The LAd[(34.5±5.0)mini, IVSTd[(9.5±1.3)mm], LVEDd[(50.2±7.7)mm], LVPWTd [(8.7±1.1)mm], LVM[(196.1±87.2)g] and LVMl[(126.5±56.4)g/m2] obviously decreased in the 6th month after self-treatment compared with prior self-treatment [(37.0±5.0), (10.2±1.5), (51.3±8.1), (9.9±1.2)mm, (230.4±95.5)g, (144.0±54.6)g/m2] and in the 3rd month after self-treatment [(35.8±5.1)mm, (10.2±1.4) ram, (51.1±8.1)nun, (9.3±1.0)mm, (219.4±82.5)g, (136.8±50.0)g/m2] (P<0.05). The results of the mitral valve flow E/A ratio and LVEF in the 3nt month after self-treatment [1.0±0.5, (59.4±13.3)%] were increased compared with the prior self-treatment[0.9±0.5, (58.1±15.6)%], and the results in the 6th month after self-treat- ment[ 1.0±0.4, (60.7±13.6)%] were further inereased compared with before, but there was no signifieant differ- ence(P0.05). Conclusions Self-treatment of Keshan disease patients can improve the heart function by pre- venting left ventrieular remodeling and reversing. Echocardiography can be used as an essential technique to evalu- ate the effect of self-treatment on Keshan disease patients.
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Objective Using flowing injection hydride generation atomic fluotometric method(FI-HG- AFM)to detect the whole blood selenium content.Methods A series of standard selenium(1,2,4,6,8.10μg/L) were dectecte by FI-HG-AFM,and the precision and accuracy of the method were observed.The selenium contents of 89 blood samples were detected by FI一HG-AFM and 2,3-diaminonaphthalene fluorometric method(DFM) respectively,and the correlation of two methods Was analyzed.Results The detection limit and recovery rate of FI-HG-AFM were 0.138μg/L and 96.4%,respectively.The relative standard deviation(RSD)was maximized at 1 μg/L(4.406%).The RSD diminished along with the increase of the concentration,and it declined to 0.512%at 10 μg/L.There wag no significant difference(t=1.7878,P>0.05)between the result8 of FI-HG-AFM[(45.9± 14.5)μg/L]and DFM[(47.5 ±13.3)μg/L],and the two methods were in highly positive correlation(r=0.8143,P< 0.01).Conclusions The whole blood selenium content call be rapidly and exactly detected by FI-HG-AFM,and the method are highly consistent with DFM.
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Objective To evaluate the self-treatment effectiveness on patients with ehwnic Keshan disease.Methods Twenty patients with chronic Keshan disease were selected from individuals with Keshan disease in Fuyu County,Heilongjiang Province.They were trained three times every three months of self management including pathogenetic condition education,general guidance,drug therapy,and they also taught how to adiust the doBe of drug according to their illness.Major symptom score,heart rate(HR),ultrasoundcardiogram (UCG)index and cardiac functional grading of these patients at basehne,after 3 months and 6 months of treatment were compared.Results The 20 patients rated their main symptoms score as(15.03 ±6.77)before self- treatrnent,and significantly decreased to(7.25±4.82)and(6.70±4.90)after 3 and 6 months treatment(P<0.01); the heart rate(HR) was (76.40±12.06) beats per minute(bpm)before self-treatment,and dramatically decreased to (69.95±12.63),(67.15±9.76)bpm after 3 and 6 months treatment(P<0.01).As for UCG detecting index,left atrial diameter(Lad)aIld left ventricular end-diastolic diameter(LVEDd)Was(37.85 ±5.23)nun and(52.49± 9.38)mm separately before self-treatment,and notablely decreased to(36.77 ±5.63),(52.15 ±9.24)mm,and (35.29±5.50),(50.81±8.88)mm respectirely after 3 and 6 months of treatment(P<0.01 or<0.05);left ventricuIar ejection fractiOII(LVEF)markedly increased(P<0.05),from(55.15±15.80)%at baseline to(57.35± 12.51)%at 3 months and(60.30±13.42)%at 6 months;there were no significant differences in mitral flow E/A ratio changes before and after treatment(P>0.05);compared with prior to the treatment.cardiac function grading was significantly better aftertreatmentfor 3 months(T=36.0,P<0.05),but not after 6 months(T=17.5,P> 0.05).Conclusions The patients'serf-treatment is effective,which we recommend to uphold and widespread.
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Objective To acquaint the situation of selenium nutrition level and coxsakievirus B(CVB) infection in Keshan disease(KD)patients in Heilongjiang province,and provide the evidence for taking the direction mell811rement$of KD contr01.Methods The subjeets were divided into three groups:disease area KD group, disease area control group,and non-disease area control group.We collected soil,food and blood,which were used to detect selenium content,and assay IgN antibody of CVB in plasma.The selenium content of blood was determined with hydride generation atomic fluorometry spectrometrie.The lgM antibody of CVB was measured with ELISA test Kit.Results Compared with those in non-disease areas,selenium content of soil,maize,wheat and bean(0.092 ±0.011,0.003±0.001,0.005±0.003,0.006±0.001)μg/L in disease area Welre significantly lower(t=17.007.8.551,15.842,12.109;all P<0.01).In disease area,the levels of blood selenium in disease area KD group(34.803.±13.302)μg/L was obviously decreased compared to disease area control group(41.235±13.571) μg/L (P<0.05).The rate of CVB infection among disease area KD group,control in disease area and control in non-disease area were 90.0%,68.3%and 45.8%.The odds ratio(OR) value of CVB infected exposure between disease area and non-disease areas was 3.957,and the 95% confidence interval(CI)Was 1.898～8.246.The OR value of CVB infected exposure between KD group and control group was 4.171,with the 95%CI of 1.298～13.404.Compared with CVB non-infection group,blood selenium of CVB infection group was,lower,but not significanfly(t=1.179, P>0.05).Conclusions Selenium deficiency and CVB infection still exist in KD area,Heilongjiang Province.
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Objective To explore the effect of associated deficiency of selenium,protein and vitamin E(VE)on the thyroid iniury and thyroid hormone metabolism of the rats in a long-term.Methods The Wistar rats were randomly divided into four groups:Group A with selenium deficiency and low protein and VE;Group B with selenium deficiency,low protein but adequate VE;Group C,adequate selenium and protein but low VE;Group D,adequate selenium.protein and VE.The rats were killed at the end of 26th week.Glutathione peroxidase(GSH-Px)activity in the rat blood and type I 5'-deiodinase activity of the rat liver were determined.The content of triiodothyronine(T3),tetraiodothyronine(T4),thyrotropic-stimulating hormone(TSH),activated oxygen(ROS)and malonaldehyde(MDA)were detected in serum. The changes of thyroid histopathology were observed under light microscope.Results ①The interactive effect of selenium+protein and VE was not significant on GSH-Px and ID I activity(F=0.003,0.871,P＞0.05),but it was significant on MDA and ROS content(F=13.057,6.706,P＜0.05 or＜0.01). ②Selenium+protein and VE could influence T3 and T4 content(F=431.977,28.271,6.570,41.419,P＜0.05).The interactive effect of selenium±protein and VE was not significant on T3 and T4 content(F=0.871,0.136,P＞0.05).Whether in the condition of low selenium and protein or supplementary,T4 contents of supplementary VE group[(79.095±12.199),(64.392±6.261)μg/L]were respectively higher than the low VE group[(61.068±6.648),(44.176±7.090)μg/L],the difference being statistically significant(t = 3.670,6.045, P ＜ 0.01). In the condition of low VE, T3[(0.718 ± 0.079)μg/L] and T4[(44.176 ±7.090)μg/L] content of supplementary selenium and protein group was lower than that in the low selenium and protein group[ (0.966 ± 0.156), (61.068 ± 6.648)μg/L], the difference being statistically significant (t = 4.568,4.916, P ＜0.01 ). With supplementary VE, T4 content of supplementary selenium and protein group[ (64.392 ± 6.261 )μg/L] was lower than that in the low selenium and protein group [(79.095 ± 12.199)μg/L], the difference being statistically significant (t = 3.033, P ＜ 0.01 ). ③Degeneration and necrosis of follicular epithelial cell were induced by diet of low selenium, protein and VE, which could be relieved by supplymentary VE. The sparseness of intracavitary glue was observed occationally in the supplementary selenium and protein but low VE group. Conclusions Long-term deficiency of selenium, protein and VE results in the decrease of the selenoenzymes of rats, which causes accumulation of the oxidative products, as well as thyroid pathological injury and thyroid hormone metabolism disorder, but supplement of adequate VE can reduce the oxidative damage in rats having low selenium and protein diet.
