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1.
Braz. j. med. biol. res ; 54(2): e9869, 2021. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS | ID: biblio-1142585

ABSTRACT

Severe blockage in myeloid differentiation is the hallmark of acute myeloid leukemia (AML). Trdmt1 plays an important role in hematopoiesis. However, little is known about the function of Trdmt1 in AML cell differentiation. In the present study, Trdmt1 was up-regulated and miR-181a was down-regulated significantly during human leukemia HL-60 cell differentiation after TAT-CT3 fusion protein treatment. Accordingly, miR-181a overexpression in HL-60 cells inhibited granulocytic maturation. In addition, our "rescue" assay demonstrated that Trdmt1 3′-untranslated region promoted myeloid differentiation of HL-60 cells by sequestering miR-181a and up-regulating C/EBPα (a critical factor for normal myelopoiesis) via its competing endogenous RNA (ceRNA) activity on miR-181a. These findings revealed an unrecognized role of Trdmt1 as a potential ceRNA for therapeutic targets in AML.


Subject(s)
Humans , Leukemia, Myeloid, Acute/genetics , MicroRNAs/genetics , DNA (Cytosine-5-)-Methyltransferases/genetics , Cell Differentiation , HL-60 Cells
2.
Article in Chinese | WPRIM | ID: wpr-872883

ABSTRACT

Objective:To study the protective effect of modified Sinisan on liver injury caused by chronic unpredictable mild stress (CUMS) combined with solitary nutrition method based on a depression model. Method:Forty-eight SD male rats were randomly divided into normal group, depression model group, fluoxetine group (1.58 mg·kg-1·d-1), and low, medium and high-dose modified Sinisan (3.67, 7.34, 14.68 g·kg-1·d-1) groups, with 8 rats in each group. After the administration of the model, the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST) in serum and the levels of inflammatory factors tumor necrosis factor-α(TNF-α), interleukin(IL)-1β, and IL-6 in liver tissues were measured by a biochemical kit method. Western blot was used to detect the expressions of cysteine aspastic acid-specific protease-3 (Caspase-3) and cleaved Caspase-3 in liver cells after intervention with modified Sinisan. Result:Compared with the normal group, the biochemical test showed that the levels of ALT and AST in the serum of the model group were significantly increased (P<0.05), and the levels of inflammatory factors TNF-α, IL-1β and IL-6 in liver tissue significantly increased (P<0.05). Compared with the model group, low, medium-dose modified Sinisan groups significantly reduced serum ALT, AST and inflammatory factors TNF-α, IL-1β and IL-6 levels (P<0.05). Western blot showed that compared with the normal group, the protein expression of Caspase-3 in model rat liver cells decreased, while the protein expression of cleaved Caspase-3 increased(P<0.05,P<0.01). Compared with the model group, low, medium-dose modified Sinisan groups could up-regulate Caspase-3 protein expression, and down-regulated the expression of cleaved Caspase-3 protein(P<0.05,P<0.01). Conclusion:Chronic stress-induced depression model can cause liver damage. Modified Sinisan can have the hepatoprotective effect by regulating the levels of various physiological, biochemical and inflammatory factor indicators, and inhibiting liver cell apoptosis.

3.
Article in Chinese | WPRIM | ID: wpr-773712

ABSTRACT

To investigate the effects of Shugan Hewei Decoction and its active substance fractions on behavior and neurotransmitter levels in hypothalamus of depression model rats,and preliminarily explore its possible mechanism. Male SD rats were randomly divided into blank control group,model group,fluoxetine( positive control) group,Shugan Hewei Decoction high and low dose groups,high and low dose groups of three different substance fractions. After 3 weeks' CUMS and social isolation to induce models,intragastrical administration lasted for 7 d. Behavioral experiments( sucrose consumption test,open-field test,and forced swimming test) were then performed to evaluate the depression status of rats. Several neurotransmitters in hypothalamus of rats were determined by LC-MS/MS method,including dapamine( DA),norepinephrine( NE),serotonin( 5-HT),5-indoleacetic acid( 5-HIAA),γ-aminobutyric acid( GABA),and glutamic acid( Glu). As compared with the blank control group,the sucrose consumption was reduced( P<0. 01); the total distance and the number of crossing the central area were also significantly reduced( P< 0. 01,P< 0. 01),while the resting time increased significantly( P<0. 01); the forced swimming time was significantly prolonged( P<0. 01); DA,5-HT,NE,5-HIAA and GABA levels in hypothalamus were significantly reduced( P < 0. 01),while Glue level was significantly increased( P < 0. 01) in model group. As compared with the model group,all the above indexes had changes in fluoxetine group,Shugan Hewei Decoction whole recipe groups,volatile oils group,polysaccharides group,and terpenoids group( P<0. 01 or P<0. 05). Shugan Hewei Decoction whole recipe and its active substance fractions can improve the behavior of depression model rats and may exert anti-depression effects by regulating the content of neurotransmitters in the hypothalamus.


Subject(s)
Animals , Chromatography, Liquid , Depression , Drug Therapy , Drugs, Chinese Herbal , Pharmacology , Hypothalamus , Chemistry , Male , Neurotransmitter Agents , Chemistry , Random Allocation , Rats , Rats, Sprague-Dawley , Tandem Mass Spectrometry
4.
Article in Chinese | WPRIM | ID: wpr-777469

ABSTRACT

To study the antidepressant effect of Shugan Hewei Tang on chronic stress depression model rats, and select the effective substance fractions. Several male SD rats were randomly divided into 17 groups: blank control group, model group, positive control group(fluoxetine), Shugan Hewei Tang high and low dose groups, 6 high and low dose groups of different substance fractions. After modeling for 3 weeks and administration for 1 week, the effective substance fractions were selected according to the body mass and behavioral performance of SD rats in each group; several neurotransmitters in hippocampus of rats were determined by LC-MS/MS method, including norepinephrine(NE), serotonin(5-HT), 5-indoleacetic acid(5-HIAA), r-aminobutyric acid(GABA), and glutamic acid(Glu). Behavioral results after modeling showed that as compared with the blank group, the body mass growth of the model group was significantly reduced(P<0.01); the sugar water consumption was reduced(P<0.01); the distance between the open field and the number of crossing the central area were also significantly reduced(P<0.01, P<0.01); the resting time was increased significantly(P<0.01); and the forced swimming time was significantly prolonged(P<0.01), indicating that the depression model was effective. After intragastric administration, as compared with the model group, the above detection indicators of volatile oils, total polysaccharides and terpenoid in Fluoxetine, Shugan Hewei Tang groups were all changed(P<0.05 or P<0.01). The levels of NE, 5-HT and GABA in the hippocampus of the model group were significantly lower than those in the blank group(P<0.01), and the levels of 5-HIAA and Glu were significantly increased(P<0.01). As compared with the model group, neurotransmitters of the treatment groups were changed obviously or significantly(P<0.05 or P<0.01). Shugan Hewei Tang showed obvious anti-depressant effects, and the volatile oils, total polysaccharides and terpenoids acted as the main active substances. The mechanism of anti-depression may be related to its regulation of various neurotransmitter levels in the hippocampus.


Subject(s)
Animals , Chromatography, Liquid , Depression , Drug Therapy , Disease Models, Animal , Drugs, Chinese Herbal , Pharmacology , Hippocampus , Metabolism , Male , Neurotransmitter Agents , Metabolism , Oils, Volatile , Pharmacology , Random Allocation , Rats , Rats, Sprague-Dawley , Stress, Psychological , Tandem Mass Spectrometry
5.
Article in Chinese | WPRIM | ID: wpr-661138

ABSTRACT

Objective To investigate the effect of single dose intraosseous injection of simvastatin on tumor vascular structure and function in murine breast cancer. Methods BALB/c mice and 4T1 murine breast cancer cells were used to establish a subcutaneous xenograft model. The mouse model of orthotopic breast cancer received intraosseous injection of a single dose of simvastatin (50 μg) or vehicle only. Frozen tumor tissue sections were prepared for co-immunostained with CD31 andα-SMA. Evans blue dye was injected into the tail vein to observe the vascular permeability. The expression level of HIF-1αwas detected by immunohistochemistry. Results Immunofluorescence dual staining showed that intraosseous injection of simvastatin increased the number of perivascular pericytes in the tumor vessel(P < 0. 05), Evans blue dye content showed that in vivo vessel permeability in the tumor tissue was significantly decreased(P < 0. 05), and the immunohistochemistry results showed that local hypoxic area was significantly improved. Conclusions Single dose intraosseous injection of simvastatin can promote the normalization of tumor vasculature by improving the coverage of pericytes.

6.
Article in Chinese | WPRIM | ID: wpr-660798

ABSTRACT

Objective To explore the effect of single local intraosseous injection of small dose simvastatin on the angiogenesis and cardiac function in rats after myocardial infarction. Methods Adult male Wistar rats were divided into sham operation group, myocardial infarction model group and intraosseous injection of simvastatin 0. 5 mg group ( all n=12 per group) . The left anterior descending branch of coronary artery was ligated to establish a rat model of myocardial infarc-tion. The left ventricular function was evaluated by small animal echocardiography at 4 weeks postoperatively. The rest of the rats were sacrificed, the myocardial infarct size was evaluated by TTC staining, and the myocardial neovascularization was detected by immunofluorescence staining. Results We successfully established the rat model of myocardial infarction. The echocardiography showed that the left ventricular systolic function was decreased significantly at 4 weeks after myocardi-al infarction. Intraosseous injection of simvastatin (0. 5 mg) did not improve the left ventricular function after myocardial infarction in the rats. TTC staining showed that intraosseous injection of simvastatin did not reduce myocardial infarct size. Immunofluorescence staining showed that the myocardial capillary density of simvastatin group was slightly higher than that of myocardial infarction model group, but showing no significant difference between them. Conclusions Intraosseous in-jection of simvastatin 0. 5 mg 24 hours after myocardial infarction cannot significantly promote myocardial angiogenesis, which is believed to be beneficial to the revascularization after ischemia, and thus failed to improve the cardiac function.

7.
Article in Chinese | WPRIM | ID: wpr-333411

ABSTRACT

Combined hepatocellular-cholangiocarcinoma (CHC) is a mixed tumor containing elements of both hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC).Its remarkable histological heterogeneity has been linked to putative hepatic progenitor cell (HPC) origin.However,detailed histological or phenotypic description is rarely documented.In the present study,we reassessed 68 cases previously diagnosed as hepatitis B-related CHCs by immunohistochemistry and double-fluorescence immunostaining,focusing on HPC associated phenotypic observation of intermediate area of the tumor.It was found that tumor cells showed remarkable heterogeneity in intermediate area.Tumor cells with intermediate morphology between hepatocytes and cholangiocytes were oval-shaped and small with scant cytoplasm and hyperchromatic nuclei,arranging in solid nests mostly.By Keratin 7 (K7) staining,it appeared that the nests of tumor cells represented a maturation process from the undifferentiated small cells to mature hepatocytes through the "transitional" cells.Then,these small cells were further confirmed with intermediate phenotype as HPC by exploring immature hepatocellular marker and HPC/biliary markers co-localization.In conclusion,the HPC associated trait in CHC can be interpreted by HPC origin or gain of"stemness" by dedifferentiation.It is still too soon to give a final word that it is innate or acquired signature of HPC associated trait in CHC.

8.
Basic & Clinical Medicine ; (12): 1507-1512, 2017.
Article in Chinese | WPRIM | ID: wpr-666998

ABSTRACT

Objective To prepare a protein chip for the detection of serum antibodies against several peach allergen components, and to provide a rapid and reliable method for clinical diagnosis of peach allergic disease .Methods The coding sequences of six key peach components Pru p 1, Pru p 2, Pru p 3, Pru p 4, Pru p 7 and Pru p G were inserted into pGAPZαA yeast expression vector.Then the vectors were digested by endonuclease Avr Ⅱ and electroporated into yeast SMD1168 for protein expression.After peach proteins were purified, the protein chip was prepared and used to detect the antibodies against peach allergen components in serum samples from 41 suspected patients.The sensitivity and specificity of the protein chip were verified by comparison with ImmunoCAP method .Res ults The protein chip was prepared with these proteins for detecting IgE antibodies in serum samples against these four peach allergen components.The results showed that the sensitivity of the protein chip to Pru p 1, Pru p 3 and Pru p 4 was 40%, 100%and 100%, and the specificity was 100%, 50%and 100%respectively.The total sensitivity and specificity was 86%and 96%respectively.Six serum samples from peach allergy patients were identified Pru p7 antibody positive by the protein chip.Co nclusions The sensitivity and specificity of the protein chip is comparable to ImmunoCAP.It needs less serum and so to be a potential method for the clinical diagnosis of peach allergenic disease after optimization.

9.
Article in Chinese | WPRIM | ID: wpr-658258

ABSTRACT

Objective To investigate the effect of single dose intraosseous injection of simvastatin on tumor vascular structure and function in murine breast cancer. Methods BALB/c mice and 4T1 murine breast cancer cells were used to establish a subcutaneous xenograft model. The mouse model of orthotopic breast cancer received intraosseous injection of a single dose of simvastatin (50 μg) or vehicle only. Frozen tumor tissue sections were prepared for co-immunostained with CD31 andα-SMA. Evans blue dye was injected into the tail vein to observe the vascular permeability. The expression level of HIF-1αwas detected by immunohistochemistry. Results Immunofluorescence dual staining showed that intraosseous injection of simvastatin increased the number of perivascular pericytes in the tumor vessel(P < 0. 05), Evans blue dye content showed that in vivo vessel permeability in the tumor tissue was significantly decreased(P < 0. 05), and the immunohistochemistry results showed that local hypoxic area was significantly improved. Conclusions Single dose intraosseous injection of simvastatin can promote the normalization of tumor vasculature by improving the coverage of pericytes.

10.
Article in Chinese | WPRIM | ID: wpr-658079

ABSTRACT

Objective To explore the effect of single local intraosseous injection of small dose simvastatin on the angiogenesis and cardiac function in rats after myocardial infarction. Methods Adult male Wistar rats were divided into sham operation group, myocardial infarction model group and intraosseous injection of simvastatin 0. 5 mg group ( all n=12 per group) . The left anterior descending branch of coronary artery was ligated to establish a rat model of myocardial infarc-tion. The left ventricular function was evaluated by small animal echocardiography at 4 weeks postoperatively. The rest of the rats were sacrificed, the myocardial infarct size was evaluated by TTC staining, and the myocardial neovascularization was detected by immunofluorescence staining. Results We successfully established the rat model of myocardial infarction. The echocardiography showed that the left ventricular systolic function was decreased significantly at 4 weeks after myocardi-al infarction. Intraosseous injection of simvastatin (0. 5 mg) did not improve the left ventricular function after myocardial infarction in the rats. TTC staining showed that intraosseous injection of simvastatin did not reduce myocardial infarct size. Immunofluorescence staining showed that the myocardial capillary density of simvastatin group was slightly higher than that of myocardial infarction model group, but showing no significant difference between them. Conclusions Intraosseous in-jection of simvastatin 0. 5 mg 24 hours after myocardial infarction cannot significantly promote myocardial angiogenesis, which is believed to be beneficial to the revascularization after ischemia, and thus failed to improve the cardiac function.

11.
Chinese Journal of Surgery ; (12): 293-295, 2010.
Article in Chinese | WPRIM | ID: wpr-254794

ABSTRACT

<p><b>OBJECTIVES</b>To analyze the pathogenic bacterial's distribution and the drug resistance in the upper urinary tract stones, and to provide the information for choosing suitable antibiotics.</p><p><b>METHODS</b>Stone samples were taken for culture and for drug sensitivity test in 146 patients undergoing percutaneous nephrolithotomy between April 2007 and October 2008, and the results were analyzed.</p><p><b>RESULTS</b>Pathogens presented in 72 (49.3%) patients. There were 70 (86.4%) Gram-negative bacteria strains. Escherichia coli, Pseudomonas aeruginosa and Enterobacter cloacae were the predominant bacteria, accounted for 30.9%(25 strains), 23.5% (19 strains) and 12.3% (10 strains), respectively. There were 10 (12.3%) Gram-positive bacteria strains, the predominant bacteria was Staphylococcus epidermidis (6 strains), accounting for 7.4%. And there was 1 fungi strain (1.2%). Resistance to ampicillin/sulbactam (88.7%), ceftriaxone (81.3%) and ciprofloxacin (67.5%) was most commonly found in pathogen, and the rate of resistance to amikacin, imipenem and piperacillin/tazobactam were 8.6%, 10.0%, 10.0%, respectively. Erythromycylamine, teicoplanin, SMZ-TMP, nitrofurantoin were sensitive to Gram-positive bacteria.</p><p><b>CONCLUSIONS</b>Bacterial's distribution of upper urinary tract stones are multiple, and the majority pathogen is Gram-negative bacteria. A big variant resistance is found among different bacterium. The suitable antibiotics should be chosen according to the different bacterium in the patients who underwent percutaneous nephrolithotomy.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Bacteria , Child , Child, Preschool , Drug Resistance, Bacterial , Female , Humans , Kidney Calculi , Microbiology , Male , Microbial Sensitivity Tests , Middle Aged , Retrospective Studies , Ureteral Calculi , Microbiology , Young Adult
12.
Chinese Journal of Surgery ; (12): 90-92, 2003.
Article in Chinese | WPRIM | ID: wpr-257723

ABSTRACT

<p><b>OBJECTIVE</b>To construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC).</p><p><b>METHODS</b>Poly A(+) RNA was isolated from RCC lines 786-O (tester) and renal cell (RC) lines HK-2 (driver), respectively. SSH procedure was performed according to the protocol of the PCR-Select cDNA Subtraction Kit (Clontech), and PCR products were cloned into pT-Adv vector and transformed E. coli TOP10F'. All positive clones picked out were digested and some of which were sequenced.</p><p><b>RESULTS</b>The SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0.3 to 0.9 kb. Among 50 clones sequenced randomly, 2 represented unknown genes and the other 48 derived from 36 known genes.</p><p><b>CONCLUSION</b>The quality of the SSH library of human RCC is reliable and its construction is the basis for further screening differentially expressed genes of RCC.</p>


Subject(s)
Adenocarcinoma, Clear Cell , Genetics , Cell Line, Tumor , Gene Library , Humans , Kidney Neoplasms , Genetics , Nucleic Acid Hybridization , Methods
13.
Article in Chinese | WPRIM | ID: wpr-679678

ABSTRACT

Objective To explore the approach and early effects of endovascular stent-graft deployment in the treatment of portal stenosis of cancerous thrombus.Methods Six cases with portal vein stenosis of cancerous thrombus,which caused by primary hepatic carcinoma(5 cases)and eholangiocarcinoma(1 case)and the severity of stenosis showed on contrast enhanced CT were more than 75% or occluded,were performed percutaneous transhepatie or transsplenic portography.FLUENCY~(TM) endovascular stent-graft(10 mm diameter)was placed at the position of stenosis after gastroesophageal varices embolization.Portal pressure was measured pre-and post-deployment.Results Stents were successfully placed in all patients.The average portal pressure decreased from 50.7 cm H_2O(1 cm H_2O = 0.098 kPa)to 41.3 cm H_2O after endovascular stent-graft deployment.The restenosis were found in 2 cases after one month.Haematemesis and refractory aseites appeared in one case respectively,the other 4 cases showed no significant symptoms above caused by portal hypertension.Conclusion It is safe and feasible for endovaseular stent-graft deployment in the treatment of portal stenosis of cancerous thrombus.Selecting the suitable indications,the symptoms of portal hypertension can be controlled effectively.

14.
Article in Chinese | WPRIM | ID: wpr-676204

ABSTRACT

Objective To investigate the effects of dominant-negative truncation mutant?NTCF4, lacking the N-terminal form of TCF4 gene,on biological characteristics of renal cancer cell line GRC-I and explore the molecular mechanisms.Methods GRC-I cell was transfected with pCDNA3-?NTCF4 eukary- otie expression plasmid,pCDNA3 empty vector to construct the stable cell line GRC-I/?NTCF4 and GRC-I/ Mock respectively.The morphological changes of stable cells were observed and the cells growth curve was detected through light microscope.The cellular proliferation activities were determined using the MTT assay. The protein expression of Wnt pathway downstream target gene C-Myc and Cox-2 was evaluated by immuno- cytoehemieal method and Western Blot analysis.Results After the dominant-negative?NTCF4 gene was permanently expressed,the GRC-I/?NTCF4 stable cells morphologically showed that appearance changed from circular to long-spindle shape,growth rate decreased with less karyosehisis found,malignant pheno- types reversed to normal renal tubular cells.MTT assay revealed that the proliferation activities of GRC-1/?NTCF4 cells were inhibited by 11.2%-35.5% compared with GRC-I cells (P<0.05),while the GRC- I/Mock cells have no difference with the control cells.Immunocytochemical analysis and Western Blot showed that the C-Myc and Cox-2 protein expression level of GRC-I/?ANTCF4 cells were significantly sup- pressed in comparison with that of GRC-I/Mock and GRC-I cells.Conclusions The dominant-negative truncation mutant?NTCF4 could partially inhibit the growth of renal cancer cells and down-regulate the pro- tein expression of Wnt pathway target gene C-Myc and Cox-2.These findings provide a experimental founda- tion for applying cell signal therapy to renal cell cancer by blocking the Wnt signaling pathway.

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