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Objective To observe the expression of leptin(Lep) receptor (LepR) and aquaporin 2 (AQP2) in the kidneys of obesity-related hypertensive rats ( OHR ) and to explore the mechanism of Lep resistance and water metabolic disorders in them.Methods OHR( model group) were induced by high-fat diet.Normal Wistar rats were chosen as normal control and hypertensive rats(SHR) as positive control.The serum level of triglycerides(TG), total cholesterol(TC), Lep, vasopressin ( AVP ) , angiotensinⅡ( AngⅡ) and β2-microglobulin (β2-MG ) was measured by enzyme linked immunosorbent assays ( ELISA) and renal morphology was observed by HE staining.The density of LepR and AngⅡtype 1( AT1) in the kidney was observed by immunohistochemistry.mRNA And protein expression of LepR and AQP2 in the kidney was assayed by real-time polymerase chain reaction and Western blotting.Results Compared with normal rats,the TG, TC, Lep, AVP, AngⅡand β2-MG of the model group were significantly increased (P<0.05), and protein and mRNA expression of LepR and AQP2 in the kidney were up-regulated (P<0.05).Compared with SHR group, TG, TC and Lep in serum of the model group were significantly increased (P<0.05).The concentrations of AVP,β2-MG and Lep was linearly related(R2 =0.87,R2 =0.95).Conclusion Water metabolic disorder and Lep resistance may be involved in the kidney injury of OHR, which may be one of the important pathogeneses of obesity-related hypertension.
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[ ABSTRACT] AIM:To investigate the preventive effect of Guizhi decoction on myocardial injury after chemical sympathectomy induced by 6-hydroxydopamine (6-OHDA).METHODS:Wistar rats (n=54) were randomly divided in-to 6 groups.Methycobal and Guizhi decoction ( with different proportions between Ramulus Cinnamomi and Radix paeoniae Alba at 2∶1, 1∶2 or 1∶1) were pre-administered to the rats.Immunohistochemical method was used to observe the cardiac sympathetic nerve distribution.The contents of tyrosine hydroxylase (TH), choline acetylaminotransferase (ChAT) and growth-associated protein 43 (GAP-43) in the left ventricle were measured by ELISA.The serum levels of myocardial en-zymes and morphology of myocardial tissues were also observed.RESULTS:6-OHDA successfully induced cardiac sympa-thetic denervation as the contents of TH and GAP-43 in the left ventricle declined significantly.Compared with model group, the content of TH was elevated in both methycobal group and Guizhi decoction groups, while the content of GAP-43 was elevated only in Guizhi decoction groups.The serum levels of myocardial enzymes and the histopathological changes of the cardiac tissues were deteriorated after injection of 6-OHDA, indicating that the myocardial injury was established. Methycobal and Guizhi decoction normalized the abnormal change.Guizhi decoctions at 2∶1 and 1∶1 showed the best effi-cacy.CONCLUSION:6-OHDA-induced sympathetic denervation causes myocardial injury.Guizhi decoction with the proportions between Ranulus Cinnamomi and Radix paeoniae Alba at 2∶1 and 1∶1 effectively alleviate the myocardial injury after cardiac sympathetic denervation induced by 6-OHDA.
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Objective To observe the effects of Chinese medicinal herbs with the efficacy of invigorating spleen, upbearing the clear, and tonifying spleen on the obesity degree, fat hormones, and insulin resistance in diet-induced obesity (DIO) rats;To choose better anti-obesity herbs from different Chinese medicinal herbs that can tonify spleen. Methods Among the 130 Wistar rats, 10 were chosen as the blank control group (fed with basal forage), and the remaining 120 were administered with high-fat high-nutrition forage for 13 weeks. According to weight, 50 DIO rats and 10 diet-induced obesity resistance (DIO-R) rats were obtained. DIO rats were divided into model group (normal saline), sibutramine group, invigorating spleen group (Atractylodix Rhizome and Magnoliae Officinalis Cortex), upbearing the clear group (Bupleuri Radix and Aurantii Fructus Immaturus) and medicine for tonifying qi and spleen group (Astragali Radix). All groups received gavage with corresponding drugs. Rats in the blank control group and the DIO-R group received gavage with normal saline. The basal forage was administered to rats in the blank control group, while high-fat forage was continually given to rats in the other six groups. Insulin resistance index (IRI), blood glucose, triglycerides, cholesterol, tumor necrosis factor α (TNF-α) and adiponectin were detected after blood withdrawing. TNF-α and adiponectin in the fat homogenate were examined. Results Compared with the blank control group, body weight, IRI, and cholesterol of rats in the model group significantly increased (P<0.01);adiponectin in homogenate reduced (P<0.01);serum and adipose homogenate TNF-α increased (P<0.01). Compared with model group, body weight, IRI, cholesterol of rats in DIO-R group significantly decreased (P<0.01);adiponectin in fat homogenate increased (P<0.01). Body weight and cholesterol of rats in the sibutramine group significantly decreased (P<0.01), while serum and adipose homogenates TNF-αdecreased (P<0.05, P<0.01). Body weight, IRI and cholesterol of rats in upbearing the clear group decreased (P<0.05, P<0.01);serum and adipose homogenate TNF-α decreased (P<0.05, P<0.01);adiponectin increased in fat homogenate (P<0.05). IRI, cholesterol and serum TNF-αof rats in the invigorating spleen group decreased (P<0.05, P<0.01);adiponectin in serum and adipose homogenate increased (P<0.05). BMI, blood glucose, IRI and cholesterol of rats in Astragali Radix group decreased (P<0.05, P<0.01);TNF-α decreased and adiponectin increased in serum and adipose homogenate (P<0.05, P<0.01). Conclusion Astragali Radix could reduce obesity induced by high-fat forage, and its effects on improving glucose and lipid metabolism disorder and IR are better than the other TCM groups. Its mechanism is related to decreasing TNF-α and increasing adiponectin level.
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Aim To investigate the preventive effect of Guizhi decoction on cardiac sympathetic sprouting induced by 4-Methylcatechol (4-MC) . Methods The rat models of cardiac sympathetic sprouting were in-duced by 10 mg · L-1 4-MC ( 10μg · kg-1 body weight, i. p. ) . Guizhi decoctions ( with different pro-portion between Ramulus Cinnamomi and Radix Paeon-iae Alba, 1 : 1, 1 : 2 and 2 : 1) and metoprolol were administered to the rats. Heart rate and electrocardio-gram ( ECG ) were observed, the content of norepi-nephrine (NE), growth associated protein (GAP-43), tyrosine hydroxylase ( TH ) and acetylcholine transfer-ase enzyme ( CHAT) in myocardial homogenate of left ventricular and right atrial were determined by ELISA method, and immunofluorescence assay was used to observe cardiac nerve sprouting and sympathetic distri-bution. Results 4-MC caused cardiac sympathetic sprouting and parasympathetic was not influenced. Heart rate of the model group was improved significant-ly and higher than that of the other groups . Compared with the model group, the content of NE, GAP-43 and TH in left ventricle and right atrium of the metoprolol group and Guizhi decoction group were decreased( P<0.05 ) , and the immunofluorescence result showed that the distribution of TH positive nerve was reduced sig-nificantly(P<0.05). It was demonstrated that Guizhi decoction of the proportion between Ramulus Cinnamo-mi and Radix Paeoniae Alba 1 : 1 and 1 : 2 had the best efficacy, which was similar to the efficacy of meto-prolol. Conclusion Guizhi decoction ( with the pro-portion between Ramulus Cinnamomi and Radix Paeon-iae Alba 1 : 1 and 1 : 2 ) effectively inhibits the cardi-ac sympathetic sprouting induced by 4-MC.
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BACKGROUND: There are some studies proposing cell membrane may be one of target cell regions for electromagnetic biological effects. However, reports responsible for cellular membrane permeability and cellular biological effects after electromagnetic irradiation are few. OBJECTIVE: This study was designed to investigate the correlations of low-power millimeter wave irradiation to cellular membrane permeability and apoptosis of HL60.DESIGN: Controlled experiment.SETTING: Thus study was performed at the Medical Experimental Central Laboratory, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA between November 2006 and April 2007. MATERIALS: The human leukemic cell lines HL60 were kindly provided by Medical Experimental Center of Guangzhou Military Area Command of Chinese PLA and HD-413.2HPSG 100 millimeter wave irradiation generator was developed by Xi'an Hengda Microwave Technology Development Company, China. METHODS: HL60 cells were irradiated by millimeter wave at frequency of 41.32 GHz and mean power density of 2 mW/cm2, and divided into five groups according to the irradiation time (0, 15, 30, 45, 60 minutes groups).MAIN OUTCOME MEASURES: Lanthanum tracing was used to observe intracellular and extracellular lanthanum particles distribution and evaluate the change of cellular membrane permeability; ultrastructure and morphological characteristics of HL60 cells were observed through an transmission electron microscope; Quantitative analysis of apoptotic cells was performed by in situ marking method. RESULTS: In the 45 and 60 minutes groups, a small amount of lanthanum particles in the cytoplasm, swollen mitochondria, expanded rough endoplasmic reticulum, and obvious apoptosis were detected by ultrastructure observation. TUNEL staining showed, compared with the 0 minute control group, the apoptosis rate showed a trend of elevation in all the irradiation groups, particularly in the 45 and 60 minutes groups. CONCLUSION: Low-power millimeter wave irradiation on HL60 cells can lead to an increase in cellular membrane permeability of HL60, which may be one of the primary causes for promoting apoptosis and producing other biological effects.
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During the electroporation cell membrane conductivity increases dramatically, so the analytical model for calculating the transmembrane voltage (TMV) induced on an unpermeabilized cell membrane in suspensions can not be used in this case. An analytical model for the TMV induced on a permeabilized cell membrane in suspensions exposed to a high voltage DC pulse field was built. First, a permeabilized cell was replaced by a sphere cell having the equivalent conductivity. Then the average field inside the permeabilized cell suspensions was calculated according to the effective medium theory. Finally, based on the analytical solution for the TMV on a single unpermeabilized cell, an analytical model for the TMV on a permeabilized cell in suspensions was derived. The model shows that the TMV depends on parameters such as the external electric field, critical angle of electroporation, cell radium, conductivity of the cytoplasm, membrane and external medium, cell arrangement and cell volume fraction.
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Humanos , Permeabilidade da Membrana Celular , Condutividade Elétrica , Campos Eletromagnéticos , Eletrofisiologia , Eletroporação , Potenciais da Membrana , Modelos Biológicos , Pulso ArterialRESUMO
Objective To observe the preventative and curative effect of Tongxinluo on microvascular inflammatory injury and vasospasm in the cardiac muscle of streptozotocin (STZ) diabetic rats. Methods STZ rats were divided into STZ group, insulin group, Tongxinluo group, with the other 8 Wistar rats as normal control. The normal group and STZ group were intragastric administered with NS, insulin group was subcutaneous injected with insulin, and Tongxinluo group was intragastric administered with Tongxinluo suspension for 8 weeks. The sections of cardiac muscle was stained in HE staining and NF-?B immunohistochemical staining. The level of adhesion molecule-1 (sICAM-1), tumor necrosis factor (TNF-?), endothelin-1 (ET-1), angiotensin-Ⅱ (AT-Ⅱ) in myocardial homogenate from intercellular, and level of AT-Ⅱ, neuropeptide Y (NPY) in plasma were determined, with sICAM-1 by enzyme-linked immunosorbent assay and the others by radioimmunoassay. The sections stained by NF-?B immunohistochemical staining were image analyzed using computer. The extent of its expression was evaluated by the mean of the density ratio of the positive side. Results Compared with normal rats, the content of sICAM-1, TNF-?, ET-1, AT-Ⅱ and the expression of NF-?B in the myocardial homogenate of STZ rats was increased (P0.05). AT-Ⅱ in plasma of STZ rats increased (P
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Objective To determine the plasma DNA level of patients with ovarian cancer and evaluate its potential clinical value.Methods Blood samples were collected from 30 ovarian cancer patients before and after surgery,and from 20 patients with ovarian benign tumor and 20 healthy women.The plasma DNA was extracted by commercial kit and detected by fluorescntmeter.Results The mean plasma DNA level in the cancer patients(25.33 ?g?L-1?17.69 ?g?L-1) was significantly higher than those in the benign tumor patients(10.28 ?g?L-1?4.80 ?g?L-1) and normal control(7.60 ?g?L-1?3.87 ?g?L-1)(P
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Objective:This study was to study effect of keeping ying and weiqi in balance on cardiac muscular collagen Ⅰ,collagen Ⅲ,angiotensinⅡ in spontaneous diabetes rats,and provide new thoughts for prevention and treatment for diabetic cardiomyopathy.Methods:40 spontaneous diabetes rats were divided into four groups at random:spontaneous diabetes control group,diformin group,Guizhi Decoction group and Huanglian Jiedu Decoction group.Except control group,each group was given intragastric administration for 12weeks.Blood glucose,collagenⅠ,collagen Ⅲ,collagenⅠ/collagen Ⅲ,angiotensin Ⅱ were determined.Euzymelinked immunosorbent assay and radio-immuniy method were used.Results:Blood glucose in Huanglian Jiedu Decoction group and diformin group decreased;Compared respectively with diformin group and control group,collagenⅠdecreased obviously and collagen Ⅲ increased signifi cantly in Guizhi Decoction group,and the collagenⅠ/collagen Ⅲ decreased.Compared with diformin group and control group,cardiac muscular homogenate AT-Ⅱ contents decreased in both of Guizhi Decoction group and Huanglian Jiedu Decoction group.Conclusion:Guizhi Decoction can decrease the content of collagen Ⅰ,AT-Ⅱ and collagenⅠ/collagen Ⅲin cardiac muscular,and increase content of collagen Ⅲ,then prevent and treat cardiac muscular collagen reconstruction and myocardial fibrosis.Huanglian Jiedu Decoction can decrease the content of AT-Ⅱ,and decrease collagenⅠ/ collagen Ⅲ,collagenⅠ slightly,and can improve cardiac muscular collagen reconstruction.Keeping ying and weiqi in balance is a new thought for prevention and treatment for diabetic cardiomyopathy.
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<p><b>OBJECTIVE</b>Identification of Rb1 mutations permits accurate genetic counseling. Characterization of the causative mutation in a large low penetrance family is likely to provide important information for tumorigenesis of retinoblastoma(RB).</p><p><b>METHODS</b>Quantitative fluorescent multiplex PCR QFM-PCR technique was used for mutation detection. Long fragment PCR, reverse transcriptase-PCR, subcloning, direct sequencing and Western blotting techniques were used for characterizing the mutation.</p><p><b>RESULTS</b>A deletion covering exons 24 and 25 of Rb gene was found in a large family with 122 members in four generations. Of the 18 carries in the family, only 11 were delivered to either unilateral or bilateral RB. The family has much low-penetrance retinoblastoma, compared with the usual, high-penetrance RB (95%). An extent of 4 kb fragment deletion was detected in genomic deletion of the mutation. cDNA and sequence data showed a 174 bp shorter than the wild type message RNA resulting in an in-frame loss of 58 residues. Further analysis demonstrated the truncated protein expression of 6000 Da shorter than wild type RB1 protein.</p><p><b>CONCLUSION</b>QFM-PCR technique has enabled the investigators to identify a large deletion covering entire exons 24 and 25 of the Rb1 gene. It is the largest deletion ever found in low penetrance RB families. The characterizations of the mutation in genomic DNA, RNA and protein have provided new evidences which enhance credence to the idea that low penetrance retinoblastoma is caused by only partially functional disable of Rb1. The data will be useful in genetic counseling, particularly significant for the unaffected carriers in RB low penetrance families.</p>
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Feminino , Humanos , Masculino , Éxons , Deleção de Genes , Expressão Gênica , Linhagem , Penetrância , Neoplasias da Retina , Genética , Retinoblastoma , Genética , Proteína do Retinoblastoma , GenéticaRESUMO
Objective Soluble serum transferrin receptor (sTfR) is a valuable indicator to diagnose iron deficiency anemia in pregnant women whose indexes will alter because of gestation. Thus it is worth to compare and analyze the levels of sTfR among the pregnant women with thalassemia, healthy pregnant women, and the pregnant women with iron deficiency anemia. Methods ?-thalassemia and ?-thalassemia were detected employing with the percentage of hemolysis and gene assay in the pregnant women randomly selected from those aged 24~30, 20~24 weeks of gestation, and with no other complications. The ratio of OD413nm of HbA2 to OD413nm of HbA was used to distinguish from the two kinds of thalassemia. Then the genetic mutation or deletion of ?-thalassemia patients and nucleotide mutation of ?-thalassemia patients were detected with thalassemia detection kit. The value of sTfR in serum of different samples was measured with ELISA kit. Results 20 pregnant women were diagnosed as thalassemia by the percentage of hemolysis higher than 63%. Among them, 10, 9 and 1 women were judged as ?-thalassemia, ?-thalassemia and ?,?-thalassemia, respectively. The mean value of sTfR was 18.614?8.464nmol/L of thelassemia women during pregnancy, which was evidently lower than 28.1 nmol/L (P
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Objective:To detect the biological activity of the recombinant human angiogenin Methods:Gel scanning was used to analyze the purity of the prepared rhANG ELISA and chicken CAM assay were used to detect specificity and biological activity The prepared rhANG was compared with the standard rhANG Results:The purity of the prepared rhANG is 96% and it could induce the angiogenesis The immunity of the prepared rhANG is better than that of the standard Conclusion:The prepared rhANG could be used for the further basic and clinical research