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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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Objective To investigate the present prevalence state of children's Kaschin-Beck disease (KBD) in Xinghai county of Qinghai province,a relative active KBD area in 2009,and to investigate their nutritional selenium level of local children and the T-2 toxin contamination level in their staple food.Methods Right hand X-ray photographs of children aged 7 - 12 in Shang,Zhong and Xia villages of Tangnaihai countryside in Xinghai county were taken.X-ray diagnosis was carried out according to the Diagnostic Criteria of Kashin Beck Disease (GB 16003-1995 ).Selected samples (children's hair,drinking water and their staple food) were collected according to X-ray film taken.Selenium contents in hair,drinking water and staple food samples were measured by 2,3-diaminonaphthalene fluorescence,and T-2 toxin in staple food sample was detected with enzyme-linked immunosorbent assay(ELISA) kits.ResultsTotal X-ray detection rate of children KBD was 12.20％(31/254) and KBD positive rate of children in Xia village was up to 14.97％(22/147),Shang village was up to 9.52％(6/63),and Zhong village was up to 6.82％ (3/44).The selenium level in children's body and outer environment was very low,namely,the selenium content in hair,drinking water,wheat and flour was (0.250 ± 0.136)mg/kg,(0.156 ± 0.046)μg/L,(0.0045 ± 0.0030)mg/kg,and (0.0067 ± 0.0116)mg/kg,respectively.The T-2 toxin level was relatively high in children's staple food,which was (78.91 ± 46.17)μg/kg in wheat and (47.47 ± 46.47)μg/kg in flour.Conclusions In relative active KBD areas of Xinghai county of Qinghai province,the children's selenium nutritional level is low,and the T-2 toxin contamination level in their staple food is relatively high,which is consistent with the distribution of local children's KBD.
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ObjectiveTo investigate the influence of recombinant thioredoxin (TRX)on apoptosis of myocardium cell in viral myocarditis of mice.MethodsTwenty-four Balb/c mice,weighting 12 - 14 g,were randomly divided into 3 groups:the control group,the virus group and the protective group,8 mice in each group.The virus group and the protective group were injected with 0.1 ml 100TCID50 Coxackie virus B3 (CVB3)intraperitoneally,and the control group was injected equal volume of saline.Therewithal the protective group was injected with TRX(2 mg/kg) by tail vein,and the virus group was injected saline the same way.After 14 days all mice were killed and hearts were taken.Changes of myocardial histopathology was observed with optical microscope,cell apoptosis was checked by TUNEL technique,and the expression of apoptosis-related proteins (Bcl-2,caspase-3)in infiltrated cell of myocardium was determined by immunohistochemistry.Results(①)Lymphocyte infiltration and necrosis were observed in survivals of the virus group,sporadic coagulation necrosis and ballooning degeneration of cells were observed in the protective group,however no myocardial lesion was found in the control group.(②)TUNEL technique showed that the positive ratio of apoptosis in the virus group and the protective group[(90.23 ± 3.63)％,(20.02 ± 2.41)％] was significantly higher than that of the control group(0.00 ± 0.00,all P ＜ 0.05),the positive ratio of apoptosis in the protective group was significantly lower than that of the virus group (P ＜ 0.05 ).(③)Immunohistochemistry showed that the expression of protein Bcl-2(+,++,+++) in the virus group and the protective group was significantly higher than that of the control group (all P ＜ 0.05).The expression of protein Bcl-2 in the protective group was significantly higher than that of the virus group(P ＜ 0.05).The expression of caspase-3 (+,++) was significantly higher in the virus group and the protective group than the control group (all P ＜ 0.05).Compared with the virus group,the expression of caspase-3 in the protective group was significantly lower(P ＜ 0.05).ConclusionTRX could inhibit cardiomyocyte apoptosis in viral myocarditis mice and the inhibition is related to regulation of apoptosis-related protein expression.
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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 study the effect of granulocyte-macrophage colony-stimulating factor(GM-CSF)on angiogenesis of rat with acute myocardial injury induced by isoproterenol(Iso). Methods A total of 60 adult male Wistar rats were randomly divided into 3 groups: normal control group, GM-CSF pretreatment group (GM-CSF group), and lso injury group, 20 rats in each group. GM-CSF group was administered recombinant human(rh)GM-CSF(5.0 μg/kg), through tail intravenous injection once a day for three days. Then the GM-CSF group and the Iso injury group were anesthetized by intraperitoneal injection of lso( 15.0 mg/kg) once a day for three days. The same dose of saline was administered in the same way to the control rats. Ten days after injection, pathological changes of myocardial damage and infarct area were examined by immunohistochemistry. The mRNA expression levels of polypeptide antigen (CD34), vascular endothelial growth factor (VEGF) and its receptor KDR/flk- 1 were measured by RT-PCR. Results The difference of myocardial necrosis area between groups was statistically significant(F=10.07, P < 0.01), in which GM-CSF group[(37.37 ± 12.98)%] was significantly less than Iso injury group[(45.51 ±14.96)%, P < 0.05]. The difference of myocardial neovascularization density index of rats between groups was statistically significant ( F = 25.54, P < 0.05 ), in which GM-CSF group [(3980.05 ± 477.22) No/mm2] was significantly higher than Iso injury group((2605.93±361.49)No/mm2,P<0.01).The differences of myocardial CD34,VEGF,KDR/flk-1 mRNA expression between groups were statistically significant(F=17.83,4.29,4.10,all P<0.01).Compared to Iso mjury group[CD34(23.85±6.06),VEGF(31.80±8.05),KDR/flk-1(30.16±8.01)]were higher in the GM-CSF group[CD34(44.04±10.13),VEGF(49A±11.59),and KDR/flk-1(46A9±7.90),all P<0.01].The expressions of myocardiM VEGF mRNA and its receptor KDR/flk-1 mRNA was positively correlated(r=0.725,R2=0.526,P<0.01).Conclusions GM-CSF prelreatmcnt increases the density ofnew blood vessels in myocardium,and reduces the Iso-induced myocardial injury in rats.
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Objective To observe the protective function of recombinant human thioredoxin(TRX) on HeLa cell injury induced by Coxsackie virus 3m(CVB3m) and to study the inhibiting effect of TRX on viral replication. Methods We infected HeLa cells with 10TCID50 CVB3m and then protected these cells with TRX (2,5,10 mg/L). The protective group of TRX, viral group, control group of TRX, and normal control group were included. Six parallel wells were set up in each group. The cell growth was observed by methyl thiazolyl tetrazolium(MTT) and contrast phase microscope. Results The results of contrast phase microscope revealed that HeLa cells were arranged tightly and polygon in normal control group; untightly, became circle and abscission in viral group; HeLa cells morphous improved by increasing TRX concentration in TRX protective group(2,5,10mg/L). MTT results of the inhibitory ratio on cell growth of TRX(2,5,10 mg/L) control group(1.2%,2.9%,6.3%) were compared with normal control group(0), there was no significant difference(all P > 0.05); and while the inhibitory ratio on cell growth of TRX(2,5,10 mg/L) protective group(32.0%,28.0%,27.0%) was compared with virus infective group(51.7%), there was a significant difference (all P < 0.05). The inhibition study of viral replication showed that compared the inhibitory ratio on cell growth of TRX(2,5,10 mg/L) protective group(26.0%,27.0%, 10.9%) with virus infective group(60.0%), there was a significant difference(all P < 0.05). In the protective groups, there was a significant difference (all P < 0.05) between low dose groups(2,5 mg/L) and high dose groups( 10 mg/L). Conclusions The recombinant TRX(2,5,10 mg/L) may alleviate HeLa cell's injury induced by virus and the construct has no significant toxicity. TRX(2,5,10 mg/L) is effective in inhibiting virus CVB3m replication.
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Objective To investigate the relationship between drinking-water type of endemic arsenicosis and adult carotid artery atherosclerosis. Methods In 2009, 285 participants aged over 40 from drinking-water type of endemic arsenism areas and 293 residents aged over 40 from control areas were investigated in Yingxian county,Shanxi province. Portable-type B mode color ultrasound was used to examine the carotid artery of all participants.The carotid atherosclerosis were diagnosed and graded through the ultrasonograms. Content of water arsenic and hair arsenic of 10 people randomly selected in every villages were detected. Results A total of 5 villages with drinkingwater type of endemic arsenicosis as observation group and 5 villages without drinking-water type of endemic arsenicosis as control group were investigated. The prevalence rates of adult carotid atherosclerosis within observation group were 35.09%(20/57), 55.74%(34/61), 38.46%(20/52), 36.51%(23/63) and 46.15%(24/52), respectively,and standardized prevalence rates were 32.5%, 33.8%, 34.9%, 46.2% and 47.3%, respectively and the prevalence rates of adult carotid atherosclerosis within control group were 18.18%(10/55), 30.77%(16/52), 20.00%(10/50),18.67% (14/75) and 21.31% ( 13/61 ), respectively; the standardize prevalence rates were 22.4%, 17.7%, 10.7%,24.6%, 18.9%, respectively. The standardize prevalence rates were higher in observation group [39.50%(113/285) ]than that in control group[39.50%(113/285), T = 26, P ＜ 0.01 ]. The severity of adult carotid atherosclerosis (composition of 4 - 7 scores ) was compared between observation group [ 17.70%(20/113 )] and control group [ 14.06% (9/64) ], and the difference was insignificant(x2 = 0.26, P ＞ 0.05). Conclusions The prevalence rate of carotid atherosclerosis in drinking-water type of endemic arsenicosis areas is higher than that of the control areas.The study provides evidence that arsenic poisoning can cause atherosclerosis.
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Objective To observe the effect of changing grain and selenium supplementation for 1-year on control of children's Kaschin-Beck disease in Qinghai province. Methods Epidemiology, clinical and right-hand X-ray examination were carried out on children aged 7 - 12 years in 2008. Patients were diagnosed and divided into 3 groups by village, control group from Xinjianping village in Guide county, changing grain group from Xiemalang village in Guide county and supplying salt with selenium and iodine group from Shanglujuan and Xialujuan villages in Xinghai county. One year before and after the treatment, right-hand X-ray photograph (including carpal bones)was taken and child hair samples were collected, selenium was detected by 2,3-diaminonaphthalene fluorescence spectrophotometry. Results After 1 year prevention and control, the detectable rate of X-ray in control group was raised from 4.88%(2/41) to 12.20%(5/41) , the detection rate in changing grain group was declined from 17.54%(10/57) to 5.26%(3/57), and from 13.51%(10/74) to 5.41%(4/74) in supplying salt with selenium and iodine group. In changing grain group, there were 10 patients, 7 cases were cured, 2 patients stable, 1 case progressed,no new case;in supplying salt with selenium and iodine group of 10 patients, 7 were cured, 3 patients stable, 1 new diagnosed case;in control group, 2 patients stable, 2 new diagnosed metaphysis cases, 1 new diagnosed metaphyseal case. Compared with control group, the difference was statistically significant between changing grain group and supplying salt with selenium and iodine group(x2 = 5.49,4.14, all P ＜ 0.05). After 1 year control and prevention,hair selenium contents in control group and changing grain group were increased from (107.15 ± 42.30), (125.30 ±40.30)μg/kg to (108.32 ± 35.67), (135.38 ± 65.24)μg/kg, the difference was statistically insignificant(t = 0.01,0.68, all P ＞ 0.05), and selenium contents in supplying salt with selenium and iodine group were obviously increased from (95.62 ± 43.42)μg/kg to (197.64 ± 97.08)μg/kg (t = 5.41, P ＜ 0.05). Conclusion Changing grain and supplying selenium can prevent and control children's Kaschin-Beck disease.
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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 investigate the trend of the national Kaschin-Beck disease condition, and provide the scientific basis for the government to make preventing strategy. Methods The data were collected from national surveillance on Kaschin-Beck disease(KBD) condition from 2000 to 2007 by restrospective method. The national X-ray detective rates and KBD condition in the eastern and western areas of China were analyzed. Results Monitoring data were collected from 189 monitoring points in 14 provinces of China, and 21 287 X-ray films were photographed of right hands of children from 2000 to 2007. X-ray detective rate was decreased significantly of national KBD from 2000 to 2007. The condition in the west of China is slightly more serious than the eastern areas, but the average of X-ray detective rate was also decreased from 21.75% to 7.30%. National condition is basically controlled. X-ray detective rate had been controlled below 5%, exept for Qinghai, Tibet and Inner Mongolia and so on. Conclusions Ninty percent of the wards have reached the controlled level according to the X diagnosis standard. In terms of the severity of the disease, most of the wards are occupied by patients with slight condition, 10% ward with moderate condition, less than 1% ward with serious condition.
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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 investigate the protective effects and molecular mechanism of peroxisome proliferate-activated receptor α(PPARα) activation on acute myocardial damage induced by isoproterenol (Iso) in rats. Methods Thirty male Wistar rats, weighting 160～180 g, were randomly divided into control group, Iso group, fenafibrate(FF) group(each n=10) according to physique quantity. Acute myocardial injury caused by Iso abdomen cavity injection induced ischemia was established and the protective effects of peroxisome proliferate-activated receptor α activation were accessed by the level of ereatine kinase(CK), lactic dehydrogenase(LDH) in serum as well as the activities of myoperoxidase(MPO) in myocardium, and the protein expressions of PPABα in myocardium by Western blot. Results The level of serum CK in control group, lso group and FF group, was (62.41±9.47),(101.71±11.05),(75.64±11.73)kU/L, respectively(F= 34.34, P<0.01). Whereas the level of serum CK in Iso group and FF group was higher than that in control group(P<0.01 or<0.05), the level of serum CK in FF group was lower than that in Iso group(P<0.01). The levels of LDH in these three groups were (5912.20±204.44), (6365.78±137.10), (6089.76±169.60) U/L, respectively(F= 17.54, P<0.01). Compared with the control group, the levels of LDH in Iso and Fir groups were significantly increased(P<0.01 or<0.05). But the level of LDH in FIr group was decreased compared with that in Iso group(P<0.01). The activities of myocardial MPO in these three groups were (1.95±0.10),(3.89±0.17),(2.49±0.19)U/g, espectively(F=391.68,P< 0.01). The activities of myocardial MPO in Iso and FF groups were higher than that in the control group (all P< 0.01), while the activities of myocardial MPO in FIr group were lower than that in lso group(P<0.01). The protein expressions of PPARα in myocardium of these three groups were 251.57±10.95,191.97±10.74,215.08±9.61, respectively(F=82.69, P<0.01). Conclusion PPARα activation by its actor FF can exert protective effects on the acute myocardial ischemia injury induced by lso in rats through inhibiting the release of inflammatory cell factors.
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Objective To observe the expressions of matrix metal proteinases-13(MMP-13) mRNA and tissue inhibitor of metal protease- 1 (TIMP- 1) mRNA and analyse the molecular mechanism of bone matrix degradation in the progress of rat subchronic fluorosis. Methods Male Wistar rats were randomly divided into two groups according to body weight, i.e. sodium floride group and control group. Rats in the sodium fluoride group were given drinking water containing 150 mg/L F-, and the animals in the control group were given tap water. The animals were bred for 24 weeks. Every 4 weeks some rats were killed. The change of obsteoclst was observed by transmission electron microscope. The expression levels of MMP-13 mRNA and TIMP-I mRNA were analyzed by RT-PCR. Results The number of lysesome and the synthesis of lysosoma enzyme in osteeclast were decreased. The expression of MMP-13 mRNA was significantly increased(t=2.29,2.41,3.07,2.52, 3.15,2.22, P<0.05) in rats treated with sodium fluoride (1.87±0.67,1.87±0.75,1.90±0.73,1.93±0.86,1.88±0.61,1.84±0.53), compared with control group(1.24±0.39, 1.19±0.27,1.07±0.22, I. 15 ~ 0.17, 1.17±0.18, 1.20±0.62). The expression of TIMP-1 mRNA was significantly increased (t=2.69,2.19,2.68,2.46,2.43,2.96, P<0.05) in rats treated with sodium fluoride(1.89±0.77,1.70±0.85,1.61±0.82,1.81±0.84,1.70±0.74, 2.06±0.96), compared with control group (1.07±0.39,0.87±0.49,0.71±0.48,0.99±0.43,0.95±0.46,0.89±0.57). Conclusion High dose fluoride might persistently induce the expressions of MMP-13 mRNA and TIMP-1 mRNA and may be involved in bone turnover.