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Objective:To investigate the expression of hippocampal synapse-related proteins including synaptophysin (SYN), postsynaptic density protein 95 (PSD95) and growth-associated protein 43 (GAP43) in rats with epilepsy accompanied by depression.Methods:The 3-month-old female clean grade SD rats were selected for the experiment.Lithium chloride pilocarpine was used to establish an epileptic rat model. Rats with successful epilepsy models were divided into epileptic depressive group (EWD group)and epileptic group with 10 in each group based on whether they were accompanied by depression. Furthermore, ten rats with matched body mass were taken as the depressive group and 10 were taken as control group. As for the depressive group rats, chronic unpredictable mild stress stimulation combined with orphanage was adopted to establish a model of depression.The depressive behaviors of rats were evaluated by body mass, sucrose preference test and open field test. Immunohistochemical staining and Western blot were used to detect the expression of SYN, PSD95 and GAP43 proteins in rat hippocampal tissue. SPSS 17.0 software was used for data statistical analysis, repeated measurement ANOVA was used for behavioral results, one-way ANOVA was used for inter group comparison of protein expression data, and LSD test was used for further pairwise comparison.Results:As for the body mass, there was significant interaction effect between the time and group among the 4 groups ( F=7.33, P<0.01). On the 8th day and the 29th day, the body weight of rats in the EWD group and the depressive group were lower than those in the epilepsy group (all P<0.05). The body weight of rats in the EWD group on the 29th day was lower than that on the first day ( P<0.05). As for the sucrose preference rates, there was significant interaction effect between the time and group among the 4 groups( F=2.67, P<0.05). The sucrose preference rate of EWD group on the15th and 29th day were lower than that on the first day (both P<0.05). The results of the open field test showed that the interaction effects of the number of vertical standing times( F=2.74) and the number of horizontal movement lattices ( F=1.76) both were not significant (both P>0.05), but both the time effect and group effect were significant (vertical standing times: Ftime=4.35, P<0.05, Fgroup=25.64, P<0.01; horizontal movement lattices: Ftime=12.75, P<0.01, Fgroup=21.37, P<0.01). The immunohistochemical results showed that there was a statistically significant difference in the number of positive cells expressing synaptic proteins SYN, PSD95 and GAP43 among the four groups of rats ( F=93.85, 58.66, 98.84, all P<0.05). The numbers of positive cells of SYN (11.73±4.30), PSD95 (24.47±7.58) and GAP43 (9.40±3.50) in the epilepsy group were lower than those in the control group ((51.00±15.39), (55.60±13.17) and (29.53±4.05)) (all P<0.05). The numbers of positive cells of SYN (5.80±3.53), PSD95 (12.87±4.03) and GAP43 (5.33±3.50) in the EWD group were lower than those in the depressive group ((11.33±3.22), (48.13±12.69) and (15.47±5.21) )(all P<0.05). Western blot results showed that there were statistically significant differences in the expression of synaptic proteins SYN, PSD95 and GAP43 among the four groups of rats ( F=13.19, 9.38, 16.80, all P<0.05). The expression levels of SYN, PSD95 and GAP43 in the epilepsy group were lower than those in the control group (all P<0.05). The expression levels of SYN, PSD95 and GAP43 in the EWD group were lower than those in the epilepsy group and the depressive group (all P<0.05). Conclusion:The low expression of SYN, PSD95 and GAP43 proteins in the hippocampus of rats with epilepsy accompanied by depression may be related to their pathogenesis.
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OBJECTIVE: To investigate the level of formaldehyde in the air of anatomical laboratories and to explore the effect of different prevention measures on reducing the level of formaldehyde. METHODS: The level of formaldehyde in two morphological laboratories of a medical college of a university were measured with a comprehensive atmospheric sampler at different physical ventilation time,different time after spraying biological enzymes and different masks. RESULTS: The level of formaldehyde decreased with the increase of ventilation time in front,middle and back of the laboratory( P < 0. 05).The level of formaldehyde at 30 and 60 minutes after spraying bio-enzymes in three parts of the laboratory was lower than that at 0 minute in the same part of the laboratory( P < 0. 05),and lower than that in the bio-enzymes non-spraying laboratory at the same part and same time point( P < 0. 05). At 0,30 and 60 minutes ventilation time points,the level of formaldehyde in the activated carbon mask group was lower than that in the normal sampling group,the non-woven mask group and the defatted gauze mask group at the same time point( P < 0. 05). CONCLUSION: Both physical ventilation and bio-enzyme degradation can reduce indoor formaldehyde pollution. Active carbon mask can achieve good protection effect.
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OBJECTIVE:To establish a method for simultaneous determination of chlorogenic acid, baicalin and imperatorin in Biyanling tablets. METHODS:HPLC method was adopted. The separation was performed on a Waters SunFire C18 column with mobile phase consisted of methanol-0.20% phosphoric acid(gradient elution)at the flow rate of 1.0 mL/min. The detection wavelength was set at 280 nm and column temperature was 30 ℃. The sample size was 20 μ L. RESULTS:Linear ranges of chlorogenic acid,baicalin and imperatorin were 1.618-16.18 μg/mL,1.624-16.24 μg/mL and 1.762-17.62 μg/mL(r were 0.999 9),respectively. The limits of quantitation were 2.67,2.03,1.87 μg/mL,and the limits of detection were 1.34,1.06, 0.98 μ g/mL,respectively. RSDs of precision,stability and reproducibility tests were all lower than 2.0%. The recoveries were 98.01%-101.80%(RSD=1.38%,n=9),98.38%-101.40%(RSD=1.26%,n=9)and 98.06%-101.40%(RSD=1.28%,n=9), respectively. RSDs of tolerance tests were all lower than 2.0%. CONCLUSIONS:The method is simple,accurate,precise,stable, reproducible and durable. It can be used for simultaneous determination of hlorogenic acid,baicalin and imperatorin in Biyanling tablets.
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Objective To investigate the relationship between the rat depression behaviors induced by chronically unpredicted stress (CUS) with interleukin-1β (IL-1β),interleukin-6 (IL-6) protein expression and indole-2,3-dioxygenase (IDO) mRNA and protein expression.Methods Thirty male Sprague Dawley (SD)rats were divided into the control group (CG) and model group (MG).The solitary raise combined with CUS 28 d continued stimulus was used to establish the rat depression model.The open field test (OFT) and the force swimming test (FST) were used to evaluate the rat depression behaviors;qRT-PCR was used to determine the mRNA expression of IDO in hippocampus and cortex;Western blot was used to determine the protein expressions of IL-1β,IL-6 and IDO in hippocampus and cortex.Results Compared with the CG group,the OFT score in the rats of the MG group was significantly decreased(P<0.01),the immobility time in FST was significantly extended (P<0.00),the expression of IDO mRNA in hippocampus and cortex was significantly increased (P<0.01) and the protein expression of IL-1β,IL-6 and IDO in hippocampus and cortex was significantly increased (P<0.05).Conclusion The rat depression behaviors induced by CUS may be associated with the increase of intracerebral inflammatory cytokine expression and inducing IDO over-expression.
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OBJECTIVE:To establish a method for simultaneous determination of chlorogenic acid,baicalin and quercetin in Compound yuxingcao tablets.METHODS:HPLC method was adopted.The separation was performed on GL InterSustain-C18 column with mobile phase consisted of water-methanol (55∶45,V/V) at the flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm,and the column temperature was 30 ℃.The sample size was 20 μL.RESULTS:Linear ranges of chlorogenic acid,baicalin and quercetin were 1.264-12.64 μg/mL (r=0.999 9),8.704-87.04 μg/mL(r=0.999 9) and 1.212-12.12 μg/mL(r=0.999 9),respectively.The limit of quantification were 1.88,2.98,1.94 ng,respectively.The limits of detection were 0.56,0.89,0.58 ng,respectively.RSDs of precision,stability and reproducibility tests were all lower than 2.0%.The recoveries were 98.02%-101.10% (RSD=1.01%,n=9),97.48 %-101.70% (RSD=1.38%,n=9) and 98.31%-101.10% (RSD=1.02 %,n=9),respectively.CONCLUSIONS:The established method is simple,accurate,stable and reproducible,and can be used for simultaneous determination of chlorogenic acid,baicalin and quercetin in Compound yuxingcao tablets.
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OBJECTIVE:To establish a method for simultaneous determination of chlorogenic acid,baicalin and quercetin in Compound yuxingcao tablets.METHODS:HPLC method was adopted.The separation was performed on GL InterSustain-C18 column with mobile phase consisted of water-methanol (55∶45,V/V) at the flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm,and the column temperature was 30 ℃.The sample size was 20 μL.RESULTS:Linear ranges of chlorogenic acid,baicalin and quercetin were 1.264-12.64 μg/mL (r=0.999 9),8.704-87.04 μg/mL(r=0.999 9) and 1.212-12.12 μg/mL(r=0.999 9),respectively.The limit of quantification were 1.88,2.98,1.94 ng,respectively.The limits of detection were 0.56,0.89,0.58 ng,respectively.RSDs of precision,stability and reproducibility tests were all lower than 2.0%.The recoveries were 98.02%-101.10% (RSD=1.01%,n=9),97.48 %-101.70% (RSD=1.38%,n=9) and 98.31%-101.10% (RSD=1.02 %,n=9),respectively.CONCLUSIONS:The established method is simple,accurate,stable and reproducible,and can be used for simultaneous determination of chlorogenic acid,baicalin and quercetin in Compound yuxingcao tablets.
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Aim To study the role of histone acetyla-tion and its involvement in the depression-like behav-iors of rats induced by chronic unpredictable stress (CUS ). Methods Thirty male Sprague Dawley (SD ) rats were randomly divided into control group and model group.The method of solitary condition with CUS for consecutive 28 days was used to establish the rat depression model.The open-field test (OFT)and the forced swimming test (FST)were used to evaluate the depressive behaviors of rats;the real time PCR was used to detect the change of HDAC2 mRNA, and Western blot was used to determine the protein expres-sions of H3,H4,acH3 and acH4 in the prefrontal cor-tex and hippocampus of rats.Results Model group showed obvious depression-like behaviors with decrea-sing locomotive activity in OFT (P <0.01 )and in-creasing immobility time in FST (P<0.01),up-regu-lating the mRNA and protein expression of HDAC2 (P<0.0 1 ),and down-regulating the protein expression of acH3 and acH4 (P<0.01)in the prefrontal cortex and hippocampus significantly,compared with control group.Conclusion The mechanism of depressive be-haviors of rats induced by CUS may be associated with down-regulating the level of histone acetylation modifi-cation.
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@#This study aimed at investigating the correlations between antidepressive effect of valproate and improvements of NET and 5-HTT expression in depressive rats leaded by chronic mild unpredicted stress(CUMS)with solitary condition. Sixty male SD rats were divided into normal group(NG), model group(MG), valproate treated-normal group(VNG), and valproate treated-model group(VMG), randomly. The changes of depressive behaviors were evaluated by the open-field test and force swimming test. The contents of MDA, activities of SOD and CAT in serum, the mRNA and protein expression of NET, 5-HTT in hippocampus were determined by biochemical methods, Real-time PCR and Western Blot, respectively. Results showed that CUMS can significantly decrease the activities in open-field test, SOD and CAT activities in serum, expression of 5-HTT in hippocampus, and obviously increase the immobility time in force swimming test, the level of MDA and expression of NET. The treatment of valproate obviously improved the changes induced by CUMS. However, the treatment of valproate had no significant influences on behaviors of NG rats. So, it revealed that improvements of the mRNA and protein expression of NET, 5-HTT may be involved in the antidepressive effect of valproate.
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Aim To investigate the relationship be-tween the depression-like behaviors of rats induced by chronically unpredictable stress( CUS) and the protein expression of tumor necrosis factor alpha( TNF-α) , in-dole-2 , 3-dioxygenase ( IDO ) and 3-hydroxyl amino acid oxygenase( HAAO) . Methods Thirty male Spra-gue Dawley( SD) rats were randomly divided into con-trol group and model group. CUS plus solitary condi-tion were used to establish the depression model. The open field test ( OFT ) and the force swimming test ( FST ) were used to evaluate the depression-like behaviors of rats ;Western blot was used to determine the protein expressions of TNF-α, IDO and HAAO in the prefrontal cortex and hippocampus of rats. Results Model group rats showed obvious depression-like be-haviors with increasing immobile time in FST ( P <0. 01) and decreasing locomotive activity in OFT( P<0. 01 ) , and up-regulating the protein expression of TNF-α, IDO and HAAO in the prefrontal cortex and hippocampus significantly ( P <0. 05 ) , compared with control group rats. Conclusion The depression-like behaviors of rats induced by CUS may be associated with the activation of IDO-HAAO pathway mediated by TNF-α.
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Aim To investigate the role of tryptophan hydroxylase-2 (TPH2),dopa-decarboxylase (DDC)and monoamine oxidase-A(MAO-A)in depression-like be-haviors induced by chronic unpredictable stress (CUS).Methods 30 male SD rats were randomly di-vided into model group(MG)and control group(CG). Rat depression model was developed by CUS for 28 consecutive days in a solitary condition.The depres-sion-like behaviors of rats were evaluated by open-field test(OFT)and forced-swimming test(FST).The real time PCR and Western blot test were used to determine the mRNA and protein expression of TPH2,DDC and MAO-A in rat telencephalon and hippocampus.Re-sults The movement scores of rats were obviously de-creased in OFT(P<0.01 ).The immobility time was obviously increased in FST (P <0.01 ).The mRNA and protein expressions of TPH2 and DDC were de-creased significantly (P <0.01,P <0.05 )and the MAO-A mRNA and protein expressions were increased significantly(P <0.01,P <0.05 )in telencephalon and hippocampus of MG rats, when compared with those in CG rats.Conclusion The TPH2,DDC and MAO-A in rat telencephalon and hippocampus were closely related with the depression-like behaviors of rats induced by CUS.
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Aim To investigate the effect of polydatin on cardiomyocyte hypertrophy induced by high glucose (25.5 mmol·L -1 )and insulin (0.1 μmol ·L -1 ) (HGI)and its possible influence on peroxisome prolif-erator-activated receptor-β (PPARβ)/nuclear tran-scription factor-κB (NF-κB)/nitric oxide (NO)signa-ling pathway.Methods The cardiomyocyte hypertro-phy was characterized in rat primary cardiomyocytes by measuring the cell surface area,protein content,and atrial natriuretic factor (ANF)mRNA expression.The mRNA and protein expressions were measured by qRT-PCR and Western blotting,respectively.The activity of NO synthase (NOS)and NO content were measured by reagent kit through ultraviolet spectroscopy.Results HGI significantly induced cardiomyocyte hypertrophy which increased the cell surface area,protein content and ANF mRNA expression (P <0.01 ).Meanwhile, the expressions of PPARβmRNA and protein reduced while the NF-κB p65 and iNOS expressions increased significantly which occurred in parallel with rising NOS activity and NO concentration (P <0.01 ).Polydatin (0.1,1,10 μmol·L -1 )inhibited the cardiomyocyte hypertrophy induced by HGI (P <0.01 ),and re-versed the mRNA and protein expressions of PPARβ, NF-κB p65 and iNOS,and NOS activity,as well as NO content.These effects of polydatin were abolished by GSK0660 (1 μmol·L -1 ),a selective PPARβan-tagonist (P <0.05 ).Conclusion Polydatin resists HGI-induced cardiomyocyte hypertrophy,which may be mediated by PPARβup-regulation,and then NF-κB-iNOS-NO pathway inactivation.
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Aim To investigate whether chronically un-predictable stress (CUS)-induced depression-like be-haviors of rats is associated with the variant expression of tryptophan hydroxylase (TPH)and tyrosine hydrox-ylase (TH).Methods 30 male SD rats were ran-domly divided into depression model group(MG)and control group (CG),the former was established using CUS plus solitary condition for 28 d,whereas the latter was fed normally as five rats per cage without CUS. The open field test(OFT)and the sucrose preference test were used to evaluate depressive behaviors.Both mRNA and protein expressions of TPH and TH in hip-pocampus and forebrain cortex were determined by re-al-time fluorescent quantitative PCR and western blot (WB),respectively.Results MG rats showed obvi-ous depressive behaviors with much lower locomotive activity and sucrose preference than CG.Meanwhile, the mRNA and protein expressions of TPH and TH also significantly decreased in MG rats,compared with CG rats.Conclusion The depression behaviors of rats in-duced by CUS may be associated with down-regulation of TPH and TH expression.
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AIM:To investigate the role of peroxisome proliferator-activated receptor β( PPARβ)-nitric oxide (NO) signal pathway in cardiomyocyte hypertrophy induced by high glucose (25.5 mmol/L) and insulin (0.1 μmol/L) ( HGI) .METHODS: The cardiomyocyte hypertrophy was characterized in rat primary cardiomyocytes by measuring the cell surface area, protein content, and the mRNA expression of atrial natriuretic factor (ANF).The mRNA and protein ex-pression were measured by real-time PCR and Western blotting , respectively .The activity of NO synthase ( NOS) and NO content were measured by a reagent kit through ultraviolet spectroscopy .RESULTS:HGI induced profound change of hy-pertrophic morphology , and significantly increased the cell surface area , protein content and mRNA expression of ANF (P<0.01), but decreased the expression of PPARβat mRNA and protein levels (P<0.05).At the same time, the ex-pression of inducible NOS (iNOS) was obviously elevated (P<0.01), which occurred in parallel with the rising NOS ac-tivity and NO concentration (P<0.01).GW0742 (1 μmol/L), a selective PPARβagonist, inhibited the cardiomyocyte hypertrophy induced by HGI ( P<0.01 ) , and up-regulated the expression of PPARβat both mRNA and protein levels . Meanwhile, GW0742 also inhibited the increases in iNOS expression , NOS activity, and NO content induced by HGI , which were abolished by GSK0660 (1 μmol/L), a selective PPARβantagonist (P<0.01).CONCLUSION: PPARβdown-regulation and the following iNOS-NO activation are involved in the cardiomyocyte hypertrophy induced by HGI .
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Aim To investigate if anti-depressive effect of venlafaxine was associated with improving oxi-dative stress and expression of hippocampus NET and 5-HTT in rat model induced by CUS. MethodsEighty SD male rats were randomly divided into four groups:model group(MG),normal group(NG), ven-lafaxine-treated normal group ( VNG ) , and venlafax-ine-treated model group ( VMG) . VNG and VMG were given venlafaxine (23. 4 mg·kg-1 ·d-1 ) once daily;NG and MG were given the same volume solvent. Soli-tary condition with chronic unpredicted stress ( CUS ) was taken to establish rat depression model. The force swimming test was used to evaluate the behavior chan-ges of experimental rats. The malondialdehyde ( MDA) level and activity of superoxide dismutase ( SOD ) in serum were determined by biochemical methods. The mRNA and protein expressions of NET and 5-HTT in hippocampus were determined by Real-Time Reverse transcription polymerase chain reaction ( real-time RT-PCR) and Western blot ( WB) , respectively. Results Compared with NG rats, obviously increasing immo-bile time of rats in force swimming test and serum MDA level, as well as significantly decreasing SOD activity in serum was observed with clearly decreasing 5-HTT expression and elevating NET expression in hippocam-pus of MG rats. The treatment of venlafaxine distinctly suppressed changes above from CUS-induced rats. However, significant changes failed to be found in VNG rats. Conclusion The anti-depressive effect of venlafaxine may at least partly involve in improving ox-idative stress/anti-oxidative stress balance and revers-ing abnormal expression of NET and 5-HTT.
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Objective To investigate the content of heavy metals in the seafood from sea areas of Guangdong province and assess the seafood safety. Methods The seafood samples were collected from seventeen representative sea areas and two seafood markets in Guangdong province from April to August 2007. The contents of Cu,Zn,Cr,Cd and Pb were measured by ICP-MS. Single factor contamination estimation in seafood was conducted and the sea food safety was assessed. Results The contents of five heavy metals in fish were below the standard limits for harmless fishery food,and the contents of heavy metals in shellfish were higher than those of fish. The contents of Cu and Zn in some oysters were higher than the evaluation standard of heavy metal pollution for the marine animals. The contents of Cd in chlamys farreri and oysters were higher than the standard limits of harmless fishery food. Except the higher content of Cr in babylonia areolata,the contents of Cr and Pb in shellfish were below the standard limits. Conclusion As for the levels of heavy metals,Cu,Zn,Cr,Cd and Pb,in the fish from the sea areas of Guangdong province is safe for health and but the levels of the five heavy metals exceeded the safety limits in varying degrees in some shellfish.
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11.1 mmol/L were treated by 2 weeks CSII.An intravenous glucose tolerance test(IVGTT) was performed before and after CSII.The levels of fasting plasma glucose(FPG),2 hours postprandial glucose(2hPG),glycosylated hemoglobin A1c(HbA1c),first-phase secretion of insulin,the mean area under the curve(AUC) of insulin,insulin sensitivity index(ISI),insulin resistance index(Homa IR) and insulin secretion index(Homa ?) were compared.Results The excellent control of FPG and 2hPG in 20 out of 21 patients were achieved stably in 2.8?1.6 days and 7.8?1.9 days by CSII.After the treatment,FPG,2h PG and HbA1c were significantly decreased(P
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To improving the teaching quality of the pharmacology theory teaching,we adopt questionnaire to understand the students’ request of teaching method and teaching content,thus to elevate the teaching overall level by raising their ability to teach and study.
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Objctive To study the effects of traditional Chinese medicine S Kangdu mixture on functions of immune system and bone marrow in mice. Methods Animal model, which marrow and immune system were inhibited, was established by 60 Co or cyclophosphamide injection. The mixture was administrated orally. The white blood cell counts, spleen colony formed unites (CFU S), and the index of carbon clean up were measured. Results The mixture obviously improved leukopenia in peripheral induced by cyclophosphamide administration, and facilitated marrow karyocyte proliferation and formation of CFU S in 60 Co treated mice and reinforced phagocytosis of mononuclear phagocytic system in cyclophosphamide treated mice. Conclusion Traditional Chinese medicine S Kangdu mixture can significantly ameliorate immunological function and stimulate hemopoietic function in mice.
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Objective: To study the roles of the oxidative stress system and the anti-oxidative stress system in neurodegenerative processes induced by aluminum overload in mice.Methods: The neurodegenerative model of mice was established via intragastric administration of AlCl3 solution(element aluminum 400 mg/kg)once a day,with 6 days of week for 3 months.The light-dark test and morris-water-maze were used to evaluate the learning and memory functions.The activities of SOD and SOD2 as well as the contents of MDA in both cortices and hippocampi were measured.Pathomorphological changes of hippocampi were observed.Results: Aluminum overload significantly decreased capacity of passive avoidance learning and memory and spatial recognition,caused the pyknosis and neuron loss of hippocampi,and parallelly increased SOD and SOD2 activities as well as MDA content in cortex and hippocampi,with a tendency of raising first and then gradually decreasing.Conclusions: Aluminum overload markedly causes neurodegeneration and damages learning and memory function in mice,the mechanism of which may be related to the imbalance between oxidative stress and anti-oxidative stress systems in brain induced by aluminum overload.
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Objective:To investigate the role of the expression of E-cadherin(E-Cad) and tissue inhibitor of metalloproteinases-2(TIMP-2) in carcinogenesis and progression of colorectal cancer(CRC).Methods:The E-Cad and TIMP-2 expressions were studied immun ohistochemistrically in 30 specimens of colorectal adenoma and 60 specimens of CRC.Results:The expression rates of E-Cad were 87.10% in colorectal adenoma and 55.00% in CRC, the former was apparently higher than that of the later (P0.05);The significant relationship was observed between low TIMP-2 expression and Dukes stage, metastasis of lymph node and distant organs and poor prognosis.Conclusion:The detection of expression about E-Cad and TIMP-2 may be helpful to judge the malignant behavior and metastasis and prognosis with colorectal carcinoma,furthermore the expression of TIMP-2 may be used to know prognosis of patients.