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This paper aimed to study the effect of Dalbergia cochinchinensis heartwood on plasma endogenous metabolites in rats with ligation of the left anterior descending coronary artery, and to analyze the mechanism of D. cochinchinensis heartwood in improving acute myocardial ischemic injury. The stability and consistency of the components in the D. cochinchinensis heartwood were verified by the establishment of fingerprint, and 30 male SD rats were randomly divided into a sham group, a model group, and a D. cochinchinensis heartwood(6 g·kg~(-1)) group, with 10 rats in each group. The sham group only opened the chest without ligation, while the other groups established the model of ligation. Ten days after administration, the hearts were taken for hematoxylin-eosin(HE) staining, and the content of heart injury indexes in the plasma creatine kinase isoenzyme(CK-MB) and lactate dehydrogenase(LDH), energy metabolism-related index glucose(Glu) content, and vascular endothelial function index nitric oxide(NO) was determined. The endogenous metabolites were detected by ultra-high-performance liquid chromatography-time-of-flight-mass spectrometry(UPLC-Q-TOF-MS). The results showed that the D. cochinchinensis heartwood reduced the content of CK-MB and LDH in the plasma of rats to relieve myocardial injury, reduced the content of Glu in the plasma, improved myocardial energy metabolism, increased the content of NO, cured the vascular endothelial injury, and promoted vasodilation. D. cochinchinensis heartwood improved the increase of intercellular space, myocardial inflammatory cell infiltration, and myofilament rupture caused by ligation of the left anterior descending coronary artery. The metabolomic study showed that the content of 26 metabolites in the plasma of rats in the model group increased significantly, while the content of 27 metabolites decreased significantly. Twenty metabolites were significantly adjusted after the administration of D. cochinchinensis heartwood. D. cochinchinensis heartwood can significantly adjust the metabolic abnormality in rats with ligation of the left anterior descending coronary artery, and its mechanism may be related to the regulation of cardiac energy metabolism, NO production, and inflammation. The results provide a corresponding basis for further explaining the effect of D. cochinchinensis on the acute myocardial injury.
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Male , Animals , Rats , Rats, Sprague-Dawley , Dalbergia , Myocardial Ischemia , Metabolomics , Heart , Heart Injuries , Creatine Kinase, MB FormABSTRACT
This study aimed to analyze the effect of Bletilla striata polysaccharide(BSP) on endogenous metabolites in serum of tumor-bearing mice treated with 5-fluorouracil(5-FU) by untargeted metabolomics techniques and explore the mechanism of BSP in alleviating the toxic and side effects induced by 5-FU. Male BALB/C mice were randomly divided into a normal group, a model group, a 5-FU group, and a 5-FU + BSP group, with eight mice in each group. Mouse colon cancer cells(CT26) were transplanted into the mice except for those in the normal group to construct the tumor-bearing mouse model by subcutaneous injection, and 5-FU chemotherapy and BSP treatment were carried out from the second day of modeling. The changes in body weight, diarrhea, and white blood cell count in the peripheral blood were recorded. The mice were sacrificed and sampled when the tumor weight of mice in the model group reached approximately 1 g. TUNEL staining was used to detect the cell apoptosis in the small intestine of each group. The proportions of hematopoietic stem cells and myeloid progenitor cells in bone marrow were measured by flow cytometry. Five serum samples were selected randomly from each group for untargeted metabolomics analysis. The results showed that BSP was not effective in inhibiting colon cancer in mice, but diarrhea, leukopenia, and weight loss caused by 5-FU chemotherapy were significantly improved after BSP intervention. In addition, apoptotic cells decreased in the small intestinal tissues and the percentages of hematopoietic stem cells and myeloid progenitor cells in bone marrow were significantly higher after BSP treatment. Metabolomics results showed that the toxic and side effects of 5-FU resulted in significant decrease in 29 metabolites and significant increase in 22 metabolites in mouse serum. Among them, 19 disordered metabolites showed a return to normal levels in the 5-FU+BSP group. The results of pathway enrichment indicated that metabolic pathways mainly involved pyrimidine metabolism, arachidonic acid metabolism, and steroid hormone biosynthesis. Therefore, BSP may ameliorate the toxic and side effects of 5-FU in the intestinal tract and bone marrow presumably by regulating nucleotide synthesis, inflammatory damage, and hormone production.
Subject(s)
Animals , Male , Mice , Colonic Neoplasms/drug therapy , Diarrhea , Fluorouracil/adverse effects , Hormones , Metabolomics , Mice, Inbred BALB C , Polysaccharides/pharmacologyABSTRACT
Objective::To observe the effect of compound Kushen injection on the expressions of transforming growth factor-β1 (TGF-β1), drosophila mothers against decapentaplegic protein 3 (Smad3), glycogen synthase kinase-3β (GSK-3β) and β-catenin mice models with radiation-induced pulmonary injury (RIPI), in order to explore its possible mechanism of action. Method::On XStrahl precision radiation research platform for small animals (SARRP), a single 20 Gy bilateral lung field irradiation was performed to establish a mice model of RIPI. Thirty-two mice were randomly divided into normal control group, model group, compound Kushen injection group and dexamethasone injection group. The normal control group and the model group were given an equal volume of 0.9%sodium chloride solution and injected intraperitoneally for 4 weeks. The pathology of lung tissue tissues was observed by using hematoxylin-eosin (HE) staining. Immunohistochemical(IHC) was used to detect the expressions of E-cadheren and Vimentin proteins in mice lung tissues.Real-time polymerase chain reaction (Real-time PCR) was used to detect the mRNA expressions of TGF-β1, Smad3, GSK-3β and β-cateninin.Western blot was used to detect the protein expressions of TGF-β1, Smad3, GSK-3β and β-cateninin. Result::Compared with the normal group, the pulmonary coefficient of the model group was significantly decreased (P<0.01). Inflammatory cell infiltration, pulmonary interstitial edema, congestion, destruction of alveolar structure and partial alveolar atrophy were observed in the lung tissues of the model group. Compared with the model group, in the compound Kushen injection group, the levels of infiltration of lung inflammatory cells and pulmonary interstitial lesions in mice, the expression of Vimentin in lung tissues (P<0.01), and the expressions of TGF-β1, Smad3, GSK-3β and β-cateninin were significantly decreased (P<0.01), whereas the expression of E-cadheren was significantly increased (P<0.01). However, compared with the dexamethasone injection group, in the compound Kushen injection group, the pathological changes of lung tissues were similar, and the expression levels of E-cadheren, Vimentin, TGF-β1, Smad3, GSK-3β and β-cateninin were not significantly different. Conclusion::Compound Kushen injection can alleviate pulmonary fibrosis of lung in the treatment of RIPI, and the mechanism may be associated with inhibiting the mRNA and protein expressions of TGF-β1, Smad3, GSK-3β and β-catenin related to epithelial-mesenchymal transition(EMT), promoting the expression of E-cadheren, and inhibiting the expression of Vimentin, so as to inhibit the occurrence of EMT.
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Objective To investigate the mechanism and function of chemokine receptor CXCR4 and its ligand CXCL12 (CXCL12 / CXCR4) in primary hepatocellular carcinoma (PHC). Methods Western blot assay, immunohistochemistry and Real-time PCR were used to detect the protein and mRNA expressions of CXCL12/CXCR4 in 60 PHC and corresponding paracancerous tissue samples. Four kinds of hepatoma cells (Huh7, MHCC97h, HepG2 and Hep3B) and normal hepatocytes (7702) were routinely cultured, and then real-time PCR was performed to detect the mRNA expressions of CXCL12/CXCR4 in these cells to screen suitable experimental cells. CXCR4 interference plasmid (sh-CXCR4) and corresponding empty vector (sh-control) were transfected into MHCC97h to construct stable transfected cell lines. The ability of invasion, migration, and proliferation of the 2 groups of cells were detected by Tanswell invasion experiment, cell scratch test and MTT test. The stably expressed sh-control and sh-CXCR4 MHCC97h cells were taken into the subcutaneous of six nude mice, and the growth of the tumor was observed. Western blot assay was used to detect the expressions of vascular endothelial growth factor-C (VEGF-C) in sh-control and sh-CXCR4 MHCC97h cell lines and corresponding xenografts in nude mice, as the same in MHCC97h, which was transfected with CXCR overexpressed plasmid. Results (1) The results of Western blot assay, immunohistochemistry and Real-time PCR showed that the expressions of CXCL12/CXCR4 protein and mRNA were significantly higher in liver cancer tissues than those of paracancerous tissues. (2) The expression levels of CXCL12/CXCR4 mRNA were higher in Huh7, MHCC97h, HepG2 and Hep3B cells than those of 7702 cells. MHCC97h was selected as the experimental cells. The ability of invasion, migration and proliferation of MHCC97h cells transfected with sh-CXCR4 were significantly lower than those of sh-control group. Meanwhile, the growth rate of nude mice transplanted with sh-CXCR4 MHCC97h cells was also significantly lower than that of sh-control group. (3) Both in vitro and in vivo experiments showed that the expression of VEGF-C was lower in sh-CXCR4 group than that in sh-control group, and the expression of VEGF-C was obviously up-regulated after overexpression of CXCR4. Conclusion High expressions of CXCL12/CXCR4 are found in primary cancer tissues and hepatoma cells. CXCL12/CXCR4 may inhibit the proliferation and invasion of hepatoma cells by regulating the expression of VEGF-C protein.
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Abnormal enhanced transmural dispersion of repolarization (TDR) plays an important role in the maintaining of the severe ventricular arrhythmias such as torsades de pointes (TDP) which can be induced in long-QT (LQT) syndrome. Taking advantage of an in vitro rabbit model of LQT2, we detected the effects of KN-93, a CaM-dependent kinase (CaMK) II inhibitor on repolarization heterogeneity of ventricular myocardium. Using the monophasic action potential recording technique, the action potentials of epicardium and endocardium were recorded in rabbit cardiac wedge infused with hypokalemic, hypomagnesaemic Tyrode's solution. At a basic length (BCL) of 2000 ms, LQT2 model was successfully mimicked with the perfusion of 0.5 μmol/L E-4031, QT intervals and the interval from the peak of T wave to the end of T wave (Tp-e) were prolonged, and Tp-e/QT increased. Besides, TDR was increased and the occurrence rate of arrhythmias like EAD, R-on-T extrasystole, and TDP increased under the above condition. Pretreatment with KN-93 (0.5 μmol/L) could inhibit EAD, R-on-T extrasystole, and TDP induced by E-4031 without affecting QT interval, Tp-e, and Tp-e/QT. This study demonstrated KN-93, a CaMKII inhibitor, can inhibit EADs which are the triggers of TDP, resulting in the suppression of TDP induced by LQT2 without affecting TDR.
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Abnormal enhanced transmural dispersion of repolarization (TDR) plays an important role in the maintaining of the severe ventricular arrhythmias such as torsades de pointes (TDP) which can be induced in long-QT (LQT) syndrome. Taking advantage of an in vitro rabbit model of LQT2, we detected the effects of KN-93, a CaM-dependent kinase (CaMK) II inhibitor on repolarization heterogeneity of ventricular myocardium. Using the monophasic action potential recording technique, the action potentials of epicardium and endocardium were recorded in rabbit cardiac wedge infused with hypokalemic, hypomagnesaemic Tyrode's solution. At a basic length (BCL) of 2000 ms, LQT2 model was successfully mimicked with the perfusion of 0.5 μmol/L E-4031, QT intervals and the interval from the peak of T wave to the end of T wave (Tp-e) were prolonged, and Tp-e/QT increased. Besides, TDR was increased and the occurrence rate of arrhythmias like EAD, R-on-T extrasystole, and TDP increased under the above condition. Pretreatment with KN-93 (0.5 μmol/L) could inhibit EAD, R-on-T extrasystole, and TDP induced by E-4031 without affecting QT interval, Tp-e, and Tp-e/QT. This study demonstrated KN-93, a CaMKII inhibitor, can inhibit EADs which are the triggers of TDP, resulting in the suppression of TDP induced by LQT2 without affecting TDR.
Subject(s)
Animals , Rabbits , Action Potentials , Anti-Arrhythmia Agents , Pharmacology , Arrhythmias, Cardiac , Benzylamines , Pharmacology , Calcium-Calmodulin-Dependent Protein Kinase Type 2 , Metabolism , Electrocardiography , Electrophysiologic Techniques, Cardiac , Endocardium , Heart , In Vitro Techniques , Long QT Syndrome , Pericardium , Piperidines , Pharmacology , Protein Kinase Inhibitors , Pharmacology , Pyridines , Pharmacology , Sulfonamides , Pharmacology , Torsades de PointesABSTRACT
Objective To evaluate the reliability and validity of the application of condensed Chinese version of the MOS 36-item short form health survey (SF-36) in assessment of quality of life among adult patients with Kashin-Beck disease,and to provide a scientific basis in rehabilitation of the patients.Methods Four hundred and twenty seven eases of adult patients with Kashin-Back disease and 419 healthy individuals randomly selected in Kashin-Beck disease endemic areas in 8 counties of Gansu province were surveyed with the SF-36.The reliability of the SF-36 was assessed by split-half reliability and Cronbach's α coefficient and the validity through principal component factor analysis and correlation analysis,etc.The dimension scores of different people were obtained by analysis of variance and univariate t-test.Results The split-half reliability of all the 8 dimensions was greater than 0.6 and the Cronbach's α coefficient was greater than 0.8; the pearson correlate coefficients of all the items to their dimensions were greater than 0.391.SF-36 contained 8 domains and 2 summary scales in the factor analysis.The score differences of quality of life in different ages of the patients,different stages of the disease were statistically significant (all P ＜ 0.05).Conclusion The SF-36 is practical in studying the quality of life among adult patients with Kashin-Beck Disease.
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Objective To understand distribution of the endemic fluorosis areas and running status of water-improving defuoridation projects in Gausu province. Methods In 2006, Gansu province endemic fluorosis areas, the content of fluoride in drinking water was measured in villages where water was not improved, running status of delluoridation projects was investigated and the content of fluoride in drinking water were determined in villages where water was improved. Dental fluorosis and skeletal fluorosis prevalence were examined in children in identified high-fluorlde villages. The fluorine content in drinking water was determined by F-ion selective electrode, dental fluorosis of children was diagnosed using Dean method, and adults skeletal fluorosis was diagnosed according to "National Standard for Clinical Diagnosis of Endemic Skeletal Fhiorosis" (GB 16396-1996). Results Water samples were examined in 1997 villages of 26 countries, among which water fluoride content was higher than 1.0 mg/L in 598 villages, accounting for 29.94%(598/1997). All 1215 water-improving and defluoridation projects had been investigated, among which 94.90%(1153/1215) of the projects were functioning well, and intermittent and abandoned projects accounted for 2.96%(36/1215) and 2.14%(26/1215). Mean fluoride of treated water of 1084 water-improving and defluoridation projects had water fluoride content ≤ 1.0 mg/L, accounted for 90.79%(1084/1194) ; mean fluoride of water from 1068 water-improving and defluoridation projects had water fuoride content ≤ 1.0 mg/L, accounting for 91.75%(1068/1164). Total 86 390 children of 8 to 12 year-old were examined, the detectable rate of dental fluorosis was 22.47%(19 414/86 390) and 142 211 adults above 16 year-old were examined, the detectable rate of skeletal fluorosis was 4.20%(5967/142 211). Conclusions Some villages yet have water fluoride content exceeding the standard. Some projects are abandoned and running badly, leading to fluoride content exceeding the standard. In a few areas, the prevalence of children dental fluorosis and adult skeletal fluorosis still exists in Gansu province, the task of prevention and control for endemic fluorosis is still arduous. We must raise the effect of prevention and treatment of this disease.
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Objective To investigate the prevalence of endemic fluorosis and the progress acchieved through control measures in Taian County of Gansu Province so as to provide basis and technique support for endemic fluorosis control. Methods The survey was carried out according to the National Surveillance Scheme of Endemic Fluorosis. Fulfillment of control measures and the quality of water improving projects were surveyed. Samples of household water and urine in children aged 8-12 years were collected and fluoride content was detected by iron selective electrode method. Children's dental fluorosis were examined with Dean method. Results In Qinan County, water was improved in a rate of 100%. In water-improving and defluoridation projects investigated, 81.15% (99/122) projects worked well, 18.85% (23/122) projects were closed or abandoned. The number of water improvement projects monitored were 24,81,9,8,respectively in the year through 2004 to 2007, revealing 13 projects having water fluorine content(>1.0-2.0 mg/L) in 2004, 15 and 5 projects having water fluorine content higher that 1.0-2.0 and 2.0-4.0 mg/L respectively in 2005, 2 projects having water fluorine content(>1.0-2.0 mg/L) in 2006, and no projects above 4.0 mg/L. In Anfu Village, the fluorine content of source and drinking water were all below 1.0 mg/L in 2006 and 2007. The urinary fluoride content of 8-12 years old children was equal and higher than 1.5 mg/L, dental fluorosis rate was 34.47% (354/1027), and dental fluorosis index was 0.65 in 2004-2007. Conclusions Projects running out of status and excessive levels of water fluorine are frequently seen, dental fluorosis is not controlled in Taian County of Gansu Province
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Objective To evaluate the running status of the water-improving projects in Gansu Province in order to further improve the supervising mechanism and raise the efficiency of defluoridation in drinking water.Methods The water-improving projects,either being normal,or intermittent,or discarded and the cause of abnormal running in 33 counties in Gansu Province were surveyed with unified standard method.Fluoride content in water was determined by F-ion selective electrode.Results 993 water improving projects covering 3389 fluorosis villages were surveyed,682 projects worked well[68.68%(682/993)]and had supplied water with normal fluoride contents to 2174 villages[64.15%(2174/3389)]and benefited 116.56 hundred thousand populations.And the rest 311[31.32%(311/993)]projects worked abnormally or supplied unqualified fluoride water.The main causes were the exceeded lasting life,insufficiency of water resources,high fluoride content in water and unqualified administration.Conclusions Most of the water-improving projects in Gansu Province are basically running normally,but about 1/3 of the projects do not function well,so the management of the projects must be improved and consummated.
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Objective To investigate the distribution of water fluoride and the present status of water- improving delluoridation projects in the endemic fluorosis areas in Gansu Province. Methods According to "The National Technical Scheme for Endemic Disease Control in 2004" for the water improving projects, water fluoride content was determined from fluorosis villages in 34 counties of 11 cities in Gansu Province. The fluoride content in drinking water was assessed by F-ion selective electrode. Results Water fluoride content was determined in 1576 fluorosis villages of 34 counties. Water fluoride content of 7829 water samples was determined, and the fluoride content of 1891 samples was over standard. Water fluoride content was ≤ 1.00 mg/L(accounting for 75.19%) in 1185 villages and 1.00 mg/L(accounting for 24.81%) in 391 villages; the highest water fluoride content was 6.78 mg/L Nine hundred and ninety three water-improving and defluoridation projects were determined. Water fluoride content of 867 water-improving and defluoridation projects was determined; 768 projects had water fluoride content ≤1.00 mg/L(accounting for 87.67%) and water fluoride content of 108 projects was 1.00 mg/L(accounting for 12.33%),with the highest water fluoride content being 5.27 mg/L. Water-improving and delluoridation projects mostly relied on drilling a well to obtain under-grand water. Abandoned projects accounted for 30%. Conclusions In 34 counties of 11 cites(prefecture), nearly 30% of the villages had water fluoride content exceeding the standard. The situation of endemic fluorosis control is still serious in Gausu Province, countermeasures for endemic fluorosis must be carried out as soon as possible and surveillance of water-improving and defluoridation projects must be strengthened.
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<p><b>BACKGROUND</b>Hepatitis B virus (HBV) X protein (HBx) and p53 could mutually down-regulate at transcriptional level and HBx could bind with p53 protein within its transactivation domain and inhibit the function of p53 protein. In recent years, effects of arsenic trioxide (As2O3) on the expression of p53 protein have been widely studied, while little is known about the activity of p53 protein. This study was undertaken to delineate the effect of HBV X gene and As2O3 on p53 protein expression (level and activity) in HepG2 cells by small hairpin RNA (shRNA)-mediated RNA interference (RNAi) technique.</p><p><b>METHODS</b>Cell line HepG2 and cells with stable expression of HBV X gene (HepG2-X) were treated with 2 micromol/L As2O3, with corresponding untreated cells serving as controls. Cell lysates and nuclear extracts were extracted. Total level and the relative activity of p53 protein were detected by modified enzyme-linked immunosorbent assay (ELISA). HBV X gene sequence-specific shRNA expression vector (pXi-1 and pXi-2) and sequence-unrelated control (pXi-3) were transfected into HepG2-X. Single cell clone with stable expression of shRNA was selected and exposed to propagating culture. The effect of As2O3 on p53 protein expression and activity was re-observed.</p><p><b>RESULTS</b>Total p53 protein level was up-regulated and its relative activity ratio was enhanced by As2O3 in HepG2 and HepG2-X cells. The total p53 protein level induced by As2O3 was up-regulated by HBV X gene expression, while its relative activity was significantly suppressed. The suppression was removed after HBV X gene expression was repressed by shRNA.</p><p><b>CONCLUSIONS</b>As2O3 up-regulates p53 protein expression and enhance its activity. HBV X up-regulates As2O3 induced-p53 protein expression while suppresses its activity.</p>
Subject(s)
Humans , Arsenicals , Pharmacology , Cell Line, Tumor , Enzyme-Linked Immunosorbent Assay , Oxides , Pharmacology , RNA Interference , Trans-Activators , Genetics , Tumor Suppressor Protein p53ABSTRACT
OBJECTIVE@#To analyze the relationship between serum HBsAg concentration and HBV replication level in hepatitis B patients with positive serum HBsAg and HBeAg, and to explore the possibility of using serum HBsAg concentration as a marker of HBV replication level in hepatitis B patients with positive serum HBeAg.@*METHODS@#HBV DNA level and serum HBeAg, HBsAg concentration of 296 patients with positive serum HBsAg and HBeAg were quantitatively detected by real-time fluorescence quantitative PCR (FQ-PCR) and time-resolved fluoroimmunoassay (TRIFA) respectively. HBsAg concentrations were compared among patients with different HBV DNA levels, and HBV DNA levels were compared among patients with different HBsAg concentrations. The correlation between serum HBsAg concentration and DNA replication level were analyzed. The positive, negative predictive values and coincidence rates were speculated by various HBsAg concentrations.@*RESULTS@#If HBV DNA positive was defined as HBV DNA levels no less than 10(5) copy/mL, then 228(77.03%) patients were classified as HBV DNA positive. HBsAg concentration was positively correlated with HBV DNA replication level, but among groups with various DNA replication levels, HBsAg concentration showed no significant statistical difference (P>0.05). If the patients were divided into 2 groups, HBsAg concentration (180 microg/L) was served as the cutoff level, the DNA positive rate of the group with HBsAg concentration no less than 180 microg/L was significantly higher than that with HBsAg concentration less than 180 microg/L (chi(2)=3.998, P<0.05). DNA positive rates and average DNA levels showed no significant statistical differences between the 2 groups, if HBsAg concentrations other than 180 microg/L were used as the cutoff level. Positive predictive values, negative predictive values and the coincidence rates speculated by various HBsAg concentrations as cutoff values did not show any significant statistical difference in estimating HBV replication levels.@*CONCLUSION@#To some extent, serum HBsAg concentration is related to HBV DNA replication level in hepatitis B patients with positive serum HBsAg and HBeAg, but it is not feasible to use HBsAg concentration to monitor their HBV replication levels.
Subject(s)
Humans , DNA, Viral , Blood , Hepatitis B Surface Antigens , Blood , Hepatitis B e Antigens , Blood , Hepatitis B, Chronic , Virology , Virus ReplicationABSTRACT
<p><b>OBJECTIVE</b>To delineate the effects of HBV X gene and of As(2)O(3) on p53 expression and activity in HepG2 cells by shRNA-mediated RNA interference (RNAi).</p><p><b>METHODS</b>HepG2 cells and cells with stable expression of HBV X gene, HepG2-X, were treated with 2 micromol/L As(2)O(3), and the corresponding untreated cells were used as controls. Cell and nuclear lysates were extracted. Total level and the relative activity absorbance of p53 were detected by modified ELISA. HBV X gene sequence-specific shRNA expression vectors, Xi-S1 and Xi-S2, and sequence-unrelated control Xi-S3 were transfected into HepG2-X. The effect of As(2)O(3) on p53 expression and activity were retested.</p><p><b>RESULTS</b>Total p53 level was up-regulated and its relative activity ratio was enhanced by As(2)O(3) in HepG2 and HepG2-X cells. The total p53 level induced by As(2)O(3) was further up-regulated by HBX expression, while its relative activity was significantly suppressed. The suppression was removed after HBX expression was suppressed by shRNA.</p><p><b>CONCLUSION</b>As(2)O(3) could up-regulate p53 expression and enhance its activity. shRNA-mediated RNA interference is conveniently being used in studies on the effect of HBV X gene expression on p53 expression and activity. HBV X expression could up-regulate p53 gene expression level induced by As(2)O(3), while it suppressed the activity of p53.</p>
Subject(s)
Humans , Apoptosis , Arsenicals , Pharmacology , Gene Expression , Hep G2 Cells , Hepatitis B virus , Genetics , Oxides , Pharmacology , RNA, Small Interfering , Trans-Activators , Genetics , Tumor Suppressor Protein p53 , Genetics , MetabolismABSTRACT
OBJECTIVE@#To construct 2 hepatocellular carcinoma (HCC) cell models for the expression of HBV X gene with different selection characteristics.@*METHODS@#HepG2 HCC cells were infected with eukaryotic expression vectors with HBV X gene, pCEP4-X, and pcDNA3. 1 (+)-X. Single cell clone was selected by hygromycin and neomycin. After propagating culture for certain periods, the HBV X gene expression was identified by PCR, RT-PCR, and Western blot.@*RESULTS@#Single HCC cell clone with HBV X gene transferred resistant to hygromycin and neomycin was selectively cultured, and the cells could be propagated for certain periods. PCR, RT-PCR, and Western blot identified the expression of HBV X gene.@*CONCLUSION@#Two HCC cell models for the expression of HBV X gene with different selection characteristics have been successfully constructed.
Subject(s)
Humans , Carcinoma, Hepatocellular , Pathology , Cell Line, Tumor , Eukaryotic Cells , Metabolism , Genetic Vectors , Liver Neoplasms , Pathology , Models, Biological , Trans-Activators , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate and compare the therapeutic effect of umbilical cord blood stem cell transplantation (UCBSCT) or adult fresh plasma in severe viral hepatitis liver failure with/without heart damage, and to study the effect of UCBSCT on liver lesions in rats.</p><p><b>METHODS</b>83 severe hepatitis patients with/without heart damage were included in the study between January 1994 and June 2003. The patients were treated with UCBSCT or given adult plasma transfusions. The therapeutic effect was evaluated by serial determination of liver function and myocardium enzymes in all patients before and after the treatment. The model of experimental hepatic failure was constructed in SD rats by injecting carbon tetrachloride. Then, the rats were given normal saline, neonate cord blood serum or neonate cord blood stem cells respectively. The expression of human AFP and Alb in SD rat livers was detected by immunohistochemistry; and human special DNA was detected by PCR.</p><p><b>RESULTS</b>The UCBSCT group had much better effects in the improvement of liver function than the adult plasma group had, no matter whether the patients had heart damage or not. Moreover, UCBSCT can decrease heart impairment of the patients. The animal experiment demonstrated that AFP and Alb positive cells were present in the neonate cord blood stem cell group after 21 days and 1 month; human special DNA was detected by PCR in these SD rat livers.</p><p><b>CONCLUSION</b>UCBSCT displayed good therapeutic effects on severe viral hepatitis and improvement of heart injury of the patients. The rat liver immunohistochemistry indicated that neonate cord blood stem cell application can decrease the liver damage and increase hepatocellular regeneration. Human umbilical cord blood stem cells can differentiate into liver cells in acute damaged SD rat livers.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Cardiomyopathies , General Surgery , Cord Blood Stem Cell Transplantation , Hepatitis, Viral, Human , General Surgery , Liver Failure , General SurgeryABSTRACT
Objective According to Kaschin-Beck Disea se monitory standardization that had been adjusted by our country,we monitored the state of Kaschin-Beck Disease in Gansu province.Methods So as to understand change of illness,we took methods of epidemiological investigation,clinical examination and X-ray diagnosis.Results It is not detected in the clinical that patient suffered from more than I of KBD among 7～12 years old in Qingyang monitory netw ork.X-ray detectable rate is 3%,but 12 cases patients were showed in Zhangjiach uan.X-ray detectable rate is 22.22%.Conclusions Illness was showed steady state and was con trolled in Qingyang region,but illness recurred clearly in Zhangjiachuan region.