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1.
Chinese Journal of Digestion ; (12): 855-861, 2019.
Article in Chinese | WPRIM | ID: wpr-800320

ABSTRACT

Objective@#To investigate the development of hepatocyte senescence during liver fibrogenesis and to explore the effect and possible mechanism of insulin-like growth factor 1 (IGF-1) on hepatocyte senescence and liver fibrosis.@*Methods@#A total of 42 male Sprague Dawley (SD) rats were selected. Eighteen rats were induced by carbon tetrachloride (CCl4) to establish the rat model of liver fibrosis. On the day 0, six and 28 after the establishment of the model, six rats were executed respectively to analyze the liver fibrosis and hepatocyte senescence in CCl4-induced liver fibrosis rat models. Twenty-four rats were divided into control group, CCl4 group, CCl4+ lentivirus vector (LV-CTR) group and CCl4+ LV-IGF-1 group, with six rats in each group.The rats were sacrificed on the 28th day after the establishment of the model. The liver tissues were obtained and the inferior vena cava blood was collected to analyze the effect of IGF-1 overexpression on liver fibrosis and hepatocyte senescence. Analysis variance (ANOVA), least significant difference (LSD) and Dunnett T3 test were performed for statistical analysis.@*Results@#Steatohepatitis on the 6th day and early stage of hepatic fibrosis on the 28th day, which indicated the model was successfully established. The results of the effects of IGF-1 overexpression on hepatic fibrosis and hepatocyte senescence showed that on the 28th day, compared with those of control group, both the score of Ishak liver inflammation and necrosis and the score of Ishak liver fibrosis were increased in the CCl4 group, CCl4+ LV-CTR group and CCl4+ LV-IGF-1 group (0, 14.55±1.94, 15.43±2.19 and 10.29±1.47, respectively; 0, 3.51±0.51, 3.21±0.79 and 1.32±0.40, respectively). The area of liver tissues by Masson staining (0.45±0.40, 5.62±1.08, 6.03±0.65 and 2.88±1.54), SA-β-Gal staining (1.75±0.80, 4.28±1.19, 4.92±1.14, 3.11±0.79), p53 (2.02±0.81, 4.36±1.02, 4.72±0.72 and 3.58±0.70) and progerin (0.72±0.40, 4.52±1.01, 4.01±1.25 and 2.66±0.80) all were increased. The levels of serum IGF-1 all were decreased ((632.00±6.04), (503.00±40.42), (508.00±21.94) and (572.40±5.94) ng/L). However the levels of ALT all were increased ((11.20±5.97), (214.00±73.90), (245.00±76.06) and (30.00±5.00) U/L). The relative expression levels of p53 (0.58±0.06, 1.78±0.18, 1.72±0.10 and 1.23±0.22) and progerin (0.12±0.02, 0.78±0.15, 1.32±0.20 and 0.81±0.16) in the primary hepatocytes were increased. The differences were all statistically significant (F=91.674, 90.778, 32.982, 9.726, 10.640, 17.029, 103.910, 30.059, 64.707 and 97.457, all P<0.05). Compared with those of CCl4+ LV-CTR group, the score of Ishak liver inflammation and necrosis and the score of Ishak liver fibrosis were decreased in the rats′ liver tissues of CCl4+ LV-IGF-1 group, the areas of Masson staining, SA-β-Gal staining, p53 and progerin in the liver tissues were decreased, the level of serum IGF-1 was increased, the level of ALT was decreased, and the relative expression levels of p53 and progerin in primary hepatocytes both were decreased. The differences were all statistically significant (all P<0.05, respectively).@*Conclusions@#Hepatocyte senescence increases in the process of liver fibrosis induced by CCl4. Overexpression of IGF-1 may alleviate liver injury, improve hepatocyte senescence and liver fibrogenesis by regulating the nuclear p53/progerin pathway.

2.
Article in Chinese | WPRIM | ID: wpr-805805

ABSTRACT

Objective@#To evaluate the role of intestinal flora disturbance in perioperative neurocognitive disorders in aged mice.@*Methods@#Sixty SPF healthy male C57BL/6J mice, aged 18 months, were divided into 4 groups (n=15 each) by a random number table method: control group (group C), operation group (group O), operation plus lactobacillus rhamnosus group (group OL) and operation plus fecal microbiota transplantation group (group OF). Exploratory laparotomy was performed in O, OL and OF groups.In group PL, lactobacillus rhamnosus 200 μl (1×109 CFU/ml, 200 μl/day) was given by gavage once a day for 10 days starting from the end of surgery, and mice received about 0.2×109 CFU probiotics per day.In group OF, broad-spectrum antibiotic mixture (ampicillin and sulbactam 1.5 g/L, vancomycin 500 mg/L, ciprofloxacin 200 mg/L, imipenem cilastatin 250 mg/L and metronidazole 1 g/L) was added to the drinking water at 7 weeks prior to operation and replaced with sterile tap water at 72 h before operation, and fecal filtrates 200 μl was given by gavage once a day for 10 days starting the end of operation.Five mice were sacrificed at day 10 after operation in each group, and Evans blue extravasation test was used to measure the vascular permeability of jejunum and ileum.Five mice were sacrificed at day 10 after operation in each group, and the small intestinal and hippocampal tissues and orbital venous blood samples were obtained for determination of interleukin-6 (IL-6), IL-17, tumor necrosis factor-alpha (TNF-α), interferon gamma (IFN-γ), IL-4 and IL-10 levels by enzyme-linked immunosorbent assay.Five mice were selected at day 10 after operation in each group, and the Morris water maze test was used to assess the cognitive function.@*Results@#Compared with group C, the vascular permeability of jejunum and ileum was significantly increased, the levels of IL-6, IL-17, TNF-α and IFN-γ in the small intestine, peripheral blood and hippocampus were increased, the levels of IL-4 and IL-10 were decreased, the swimming distance and escape latency were prolonged, and the time spent in the target quadrant was shortened in group O (P<0.05 or 0.01), and no significant change was found in the parameters mentioned above in OL and OF groups (P>0.05). Compared with group O, the vascular permeability of jejunum and ileum was significantly decreased, the levels of IL-6, IL-17, TNF-α and IFN-γ in the small intestine, peripheral blood and hippocampus were decreased, the levels of IL-4 and IL-10 were increased, the swimming distance and escape latency were shortened, and the time spent in the target quadrant was prolonged in OL and OF groups (P<0.05).@*Conclusion@#Intestinal flora disturbance can mediate inflammatory responses in the hippocampus and cause perioperative neurocognitive disorders in aged mice.

3.
Chinese Journal of Digestion ; (12): 855-861, 2019.
Article in Chinese | WPRIM | ID: wpr-824852

ABSTRACT

Objective To investigate the development of hepatocyte senescence during liver fibrogenesis and to explore the effect and possible mechanism of insulin-like growth factor 1 (IGF-1) on hepatocyte senescence and liver fibrosis.Methods A total of 42 male Sprague Dawley (SD) rats were selected.Eighteen rats were induced by carbon tetrachloride (CCl4) to establish the rat model of liver fibrosis.On the day 0,six and 28 after the establishment of the model,six rats were executed respectively to analyze the liver fibrosis and hepatocyte senescence in CCl4-induced liver fibrosis rat models.Twenty-four rats were divided into control group,CCl4 group,CCl4 + lentivirus vector (LV-CTR) group and CCl4 + LV-IGF-1 group,with six rats in each group.The rats were sacrificed on the 28th day after the establishment of the model.The liver tissues were obtained and the inferior vena cava blood was collected to analyze the effect of IGF-1 overexpression on liver fibrosis and hepatocyte senescence.Analysis variance (ANOVA),least significant difference (LSD) and Dunnett T3 test were performed for statistical analysis.Results Steatohepatitis on the 6th day and early stage of hepatic fibrosis on the 28th day,which indicated the model was successfully established.The results of the effects of IGF-1 overexpression on hepatic fibrosis and hepatocyte senescence showed that on the 28th day,compared with those of control group,both the score of Ishak liver inflammation and necrosis and the score of Ishak liver fibrosis were increased in the CCl4 group,CCl4 + LV-CTR group and CCl4 + LV-IGF-1 group (0,14.55 ±1.94,15.43 ±2.19 and 10.29 ±1.47,respectively;0,3.51 ±0.51,3.21 ±0.79 and 1.32 ±0.40,respectively).The area of liver tissues by Masson staining (0.45 ±0.40,5.62 ± 1.08,6.03 ± 0.65 and 2.88 ± 1.54),SA-β-Gal staining (1.75 ± 0.80,4.28 ± 1.19,4.92 ± 1.14,3.11 ± 0.79),p53 (2.02 ±0.81,4.36 ±1.02,4.72 ±0.72 and 3.58 ±0.70) and progerin (0.72 ±0.40,4.52±1.01,4.01 ± 1.25 and 2.66 ± 0.80) all were increased.The levels of serum IGF-1 all were decreased ((632.00 ± 6.04),(503.00 ± 40.42),(508.00 ± 21.94) and (572.40 ± 5.94) ng/L).However the levels of ALT all were increased ((11.20 ± 5.97),(214.00 ± 73.90),(245.00 ± 76.06) and (30.00 ± 5.00) U/L).The relative expression levels of p53 (0.58 ± 0.06,1.78 ± 0.18,1.72 ± 0.10 and 1.23 ± 0.22) and progerin (0.12 ± 0.02,0.78 ± 0.15,1.32 ± 0.20 and 0.81 ± 0.16) in the primary hepatocytes were increased.The differences were all statistically significant (F =91.674,90.778,32.982,9.726,10.640,17.029,103.910,30.059,64.707 and 97.457,all P < 0.05).Compared with those of CCl4 + LV-CTR group,the score of Ishak liver inflammation and necrosis and the score of Ishak liver fibrosis were decreased in the rats' liver tissues of CCl4 + LV-IGF-1 group,the areas of Masson staining,SA-β-Gal staining,p53 and progerin in the liver tissues were decreased,the level of serum IGF-1 was increased,the level of ALT was decreased,and the relative expression levels of p53 and progerin in primary hepatocytes both were decreased.The differences were all statistically significant (all P < 0.05,respectively).Conclusions Hepatocyte senescence increases in the process of liver fibrosis induced by CCl4.Overexpression of IGF-1 may alleviate liver injury,improve hepatocyte senescence and liver fibrogenesis by regulating the nuclear p53/progerin pathway.

4.
Article in Chinese | WPRIM | ID: wpr-824615

ABSTRACT

Objective To evaluate the role of intestinal flora disturbance in perioperative neurocognitive disorders in aged mice.Methods Sixty SPF healthy male C57BL/6J mice,aged 18 months,were divided into 4 groups (n =15 each) by a random number table method:control group (group C),operation group (group O),operation plus lactobacillus rhamnosus group (group OL) and operation plus fecal microbiota transplantation group (group OF).Exploratory laparotomy was performed in O,OL and OF groups.In group PL,lactobacillus rhamnosus 200 μl (1×109 CFU/ml,200 μl/day) was given by gavage once a day for 10 days starting from the end of surgery,and mice received about 0.2× 109 CFU probiotics per day.In group OF,broad-spectrum antibiotic mixture (ampicillin and sulbactam 1.5 g/L,vancomycin 500 mg/L,ciprofloxacin 200 mg/L,imipenem cilastatin 250 mg/L and metronidazole 1 g/L) was added to the drinking water at 7 weeks prior to operation and replaced with sterile tap water at 72 h before operation,and fecal filtrates 200 μl was given by gavage once a day for 10 days starting the end of operation.Five mice were sacrificed at day 10 after operation in each group,and Evans blue extravasation test was used to measure the vascular permeability of jejunum and ileum.Five mice were sacrificed at day 10 after operation in each group,and the small intestinal and hippocampal tissues and orbital venous blood samples were obtained for determination of interleukin-6 (IL-6),IL-17,tumor necrosis factor-alpha (TNF-α),interferon gamma (IFN-γ),IL-4 and IL-10 levels by enzyme-linked immunosorbent assay.Five mice were selected at day 10 after operation in each group,and the Morris water maze test was used to assess the cognitive function.Results Compared with group C,the vascular permeability of jejunum and ileum was significantly increased,the levels of IL-6,IL-17,TNF-α and IFN-γ in the small intestine,peripheral blood and hippocampus were increased,the levels of IL-4 and IL-10 were decreased,the swimming distance and escape latency were prolonged,and the time spent in the target quadrant was shortened in group O (P<0.05 or 0.01),and no significant change was found in the parameters mentioned above in OL and OF groups (P>0.05).Compared with group O,the vascular permeability of jejunum and ileum was significantly decreased,the levels of IL-6,IL-17,TNF-α and IFN-γin the small intestine,peripheral blood and hippocampus were decreased,the levels of IL-4 and IL-10 were increased,the swimming distance and escape la tency were shortened,and the time spent in the target quadrant was prolonged in OL and OF groups (P<0.05).Conclusion Intestinal flora disturbance can mediate inflammatory responses in the hippocampus and cause perioperative neurocognitive disorders in aged mice.

5.
Chinese Journal of Anesthesiology ; (12): 1318-1321, 2018.
Article in Chinese | WPRIM | ID: wpr-745598

ABSTRACT

Objective To evaluate the role of intestinal flora disturbance in development of postoperative cognitive dysfunction (POCD) in aged mice and the relationship with regulatory T cells (Treg) and T helper cells 1/T helper cells 2 (Th1/Th2) in the small intestine.Methods Thirty-six SPF healthy male C57BL/6J mice,weighing 45-50 g,aged 18 months,were divided into 3 groups (n=12 each) using a random number table method:control group (group C),POCD group and POCD plus VSL#3 group (group PV).POCD was induced by abdominal exploration.VSL#3 probiotics was given by intragastric gavage (300 μl per time,once a day) every 24 h for 7 consecutive days starting from the end of surgery in group PV.Morris water maze test was used to assess the cognitive function at day 7 after operation.Orbital venous blood samples were collected after the end of Morris water maze test,and animals were then sacrificed and small intestine and hippocampi were removed for measurement of the percentage of CD4+ CD25+ Foxp3+Treg,TCD4+IFN-γ+Th1 and CD4+IL-4+Th2 in the lamina propria of small intestine and plasma and expression of IL-4 and IFN-γmRNA in the lamina propria of small intestine,plasma and hippocampal tissues,and IL-4 mRNA/IFN-γmRNA ratio was calculated.Results Compared with group C,the percentage of CD4+CD25+Foxp3+Treg and CD4+IL-4+ Th2 in the lamina propria of small intestine and plasma was significantly decreased,the percentage of CD4+ IFN-γ+Th1 in the lamina propria of small intestine and plasma was increased,the expression of IL-4 mRNA in the lamina propria of small intestine,plasma and hippocampal tissues was down-regulated,the expression of IFN-γ mRNA in the lamina propria of small intestine,plasma and hippocampal tissues was up-regulated,IL-4 mRNA/IFN-γ mRNA ratio was decreased,the escape latency and swimming distance were prolonged,and the time spent in the target quadrant was shortened in group POCD (P<0.05).Compared with group POCD,the percentage of CD4+ CD25+ Foxp3+ Treg and CD4+IL-4+ Th2 in the lamina propria of small intestine and plasma was significantly increased,the percentage of CD4+IFN-γ+Th1 in the lamina propria of small intestine and plasma was decreased,the expression of IL-4 mRNA in the lamina propria of small intestine,plasma and hippocampal tissues was upregulated,the expression of IFN-γmRNA in the lamina propria of small intestine,plasma and hippocampal tissues was down-regulated,IL-4 mRNA/IFN-γmRNA ratio was increased,the escape latency and swimming distance were shortened,and the time spent in the target quadrant was prolonged in group PV (P< 0.05).Conclusion Intestinal flora disturbance can promote the development of POCD in aged mice,which is related to the decreased percentage of Treg and Th1/Th2 imbalance in the small intestine.

6.
China Journal of Endoscopy ; (12): 12-15, 2017.
Article in Chinese | WPRIM | ID: wpr-618525

ABSTRACT

Objective To compare the clinical effect and safety of peroral endoscopic myotomy (POEM) with surgical therapy in treatment of esophageal achalasia. Methods 78 patients diagnosed as esophageal achalasia from January 2012 to October 2014 were enrolled in the study and divided into POME group and Heller group. There were 42 patients in POEM group and 36 patients in Heller group. The clinical symptom remission rate, LES resting pressure, Eckardt scores, complication rate, length of hospital stay and the hospitalization expenses were analyzed between the two groups. Results The patients in POEM group and Heller group both got clinical remissions after the treatment. There was no statistical difference in the rate of complication occurrence, Eckardt scores and LES resting pressure between the two groups. Patients in POEM group had shorter operation time, hospital stay and less expenses compared with the Heller group. Conclusions Compared with Heller group, the POEM group has the similar curative effect in treatment of esophageal achalasia. The POEM as a minimally invasive surgery has the advantages of less pain and trauma, shorter hospital stay, well tolerated and low cost. Therefore, the POEM is worth to be popularized and applied in treating esophageal achalasia.

7.
Article in Chinese | WPRIM | ID: wpr-484764

ABSTRACT

Objective To analyze the characteristics and clinical value of time intensity curve (TIC) of contrast-enhanced ultrasound (CEUS) in recurrent small hepatocellular carcinoma (RSHCC) and primary small hepatocellular carrcinoma (PSHCC). Methods Sixty-five cases of RSHCC (all lesions ≤3 cm) were devided into group B1 with 42 cases of RSHCC (≤2 years ) , and group B2 with 23 cases of RSHCC ( > 2 yeras ) and group A invloved 49 cases of PSHCC (all lesions ≤3 cm). Enhancement patterns in arterial, portal and delayed phase were evaluated respectively in three groups through CEUS and analytic software Sonoliver was applied to obtain quantitative features of CEUS in the region of interest. Receiver operating characteristic (ROC) was drawn and the area under curve (AUC) was calculated. Results CEUS showed hyper-enhancement difference in arte-rial phase in group B2 (72.4%) and group A (94.8%)(P′ = 0.008) showed statistical significance, but no sig-nificance was found in enhanced iso in portal phase (P = 0.078). Hypo-enhancement in the delayed phase in group B2 (75.9%), group A (96.6%) and group B1 (95.3%) (P′ = 0.003, P′ = 0.005). TIC showed HT difference (half time of descending) in B2 group, A group and B1 group (P′ = 0.007, P′ = 0.013) indicated statistical significance but RT, TTP, MTT(P = 0.319,P = 0.104, P = 0.461) showed no difference. AUC was 0.841 (half time of descending). Conclusions Enhancement patterns of CEUS (RSHCC) are related to recur-rent time . En hancement patterns of RSHCC (> 2 years ) is not typical so CEUS should be combined with quanti-tative analysis of TIC to provide reference for its treatment and prognosis.

8.
Chinese Journal of Radiology ; (12): 410-413, 2015.
Article in Chinese | WPRIM | ID: wpr-467507

ABSTRACT

Objective To investigate the value of dynamic contrast?enhanced MRI (DCE?MRI) in the differential diagnosis of glioblastoma and brain metastases. Methods Twenty patients with high grade gliomas and 20 cases patients with brain metastases proved by surgery and pathology were collected, and patients were examined with conventional MRI and DCE?MRI preoperatively. The ROIs were manually placed in solid parts of the tumors and their surrounding tissues to calculate Ktrans, Kep and Ve values. The Ktrans, Kep and Ve values differences for the solid part and surrounding tissues of the two brain tumors were compared by two independent sample t test. The correlation between Ktrans of the solid parts of the two brain tumors and Ktrans, Kep and Ve values of their surrounding tissues were studied by Pearson correlation analysis. Results The Ktrans, Kep and Ve values of glioblastoma were(0.258 ± 0.063)min-1,(0.398 ± 0.082)min-1, 0.632±0.084, the Ktrans, Kep and Ve values of brain metastases were(0.233±0.053)min-1,(0.357±0.042)min-1, 0.672±0.113. There were no significant differences between the glioblastoma and brain metastases for Ktrans, Kep and Ve values(t=-1.354,-1.982, 1.276, all P>0.05). The Ktrans, Kep and Ve values of surrounding tissues of glioblastoma were(0.093±0.032)min-1,(0.411±0.089)min-1, 0.107±0.021, the Ktrans, Kep and Ve values of surrounding tissues of brain metastases were(0.033±0.010)min-1,(0.204±0.045)min-1, 0.069±0.017. The Ktrans, Kep and Ve values of surrounding tissues between glioblastoma and brain metastases had significant difference (t=-7.978,-9.303,-6.203, all P0.05). Conclusion The DCE?MRI can quantitatively display the microvascular permeability and accurately evaluate the damage of blood?brain barrier of glioblastoma and brain metastases, which has an important value in studying biological characteristics and differential diagnosis of the two brain tumors.

9.
Cancer Research and Clinic ; (6): 230-233,246, 2011.
Article in Chinese | WPRIM | ID: wpr-578534

ABSTRACT

Objective To identify,construct and express scFv CD133,verify its biological function.Methods VL and VH were isolated from hybridoma of mAb CD133 by using antibody engineering technology.Its DNA sequencing and CDR were determined.scFv CD133 was then cloned into pET32a,transformed into Origami,induced by IPTG,purified by Ni2+-NTA His resin.Its affinity and specificity were tested by NH4SCN elution and ELISA.Results The size of VL and VH of scFv CD133 was 339 bp and 342 bp,which coded 113 and 114 amino acid separately.Its VL belonged to mouse Igκ chain and VH belonged to mouse IgG heavy chain subtype I.The molecular weight of scFv CD133 was about 27 × 103 which was testified by SDSPAGE and Western blot.Its affinity and specificity were also verified.Conclusion scFv CD133 has been successfully constructed and expressed in Origami,which could supply basis for target therapy of CD+133 cancer stem cell.

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