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Objective To investigate the clinicopathological characteristics and prognosis of well-differentiated rectal neuroendocrine tumor(RNET).Methods A retrospective analysis was conducted using the clinical data from 83 patients with well-differentiated RNET from August 2017 to December 2021,including clinical manifestations,endoscopy,endoscopic treatment,postoperative complications,postoperative pathology,follow-up and prognosis.Pathological results according to the 2019 World Health Organization(WHO)Classification of digestive system tumors,83 patients were divided into G1 stage group(72 cases)and G2 stage group(11 cases);Based on the number of tumors in the patient,83 patients were divided into two groups:single RNET group(77 cases)and multiple RNET group(6 cases),the expressions of chromogranin A(CgA),synapsin(Syn)and CD56 were compared among different groups.Results Based on pathological findings in the group,G1 stage group CgA positive rate was significantly higher than that of G2 stage group,the difference was statistically significant(χ2 = 4.23,P = 0.040);Based on the number of tumors,multiple RNET group CgA positive rate was significantly higher than that of single RNET group,the difference was statistically significant(χ2 = 5.74,P = 0.017).It was no significant difference in Syn and CD56 between the two groups(P＞0.050).Conclusion Well-differentiated RNET has no specific clinical manifestations.It is mostly isolated in G1 stage and single RNET.ESD is safe and has a good prognosis,the positive rate of CgA is higher in G1 stage patients,and the positive rate of CgA is higher in patients with multiple RNET.
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Objective:To investigate Helicobactor pylori (H. pylori) infection status and interfamilial transmission pattern in Zhengzhou area. Methods:A cross-sectional study was conducted from September 2020 to march 2021, among 731 individual from 266 families randomly selected from 9 communities of Zhengzhou area. H. pylori infection status was determined by serum antibody tests, and 13C-urea breath test was performed in the previously eradicated population to clarify the current infection status. The individual and familial infection rate, infection status for couples and children and adolescent were analyzed. Results:Among 731 individuals from 266 families, 397 of them were H. pylori positive. The individual infection rate was 54.31% (397/731); among infected individuals 77.83% (307/397) were infected with type Ⅰ strain, 22.67% (90/397) were infected by type Ⅱ strain. Annual household income ( χ2=0.419, 0.410, 0.213, all P>0.05), smoking history (χ 2=0.071, P>0.05), drinking history ( χ2=0.071, P>0.05), dining place ( χ2=0.009, P>0.05), gastrointestinal symptoms ( χ2=0.047, P>0.05), family history of gastric disease ( χ2=0.069, P>0.05), and history of gastric cancer ( χ2=0.004, P>0.05) had no significant differences between H. pylori-positive and -negative groups, but the infection rate in individuals with higher education level was lower ( χ2=4.449, P<0.05). The infection rate was significantly higher in≥18 age groups compared with<18 age groups ( χ2=6.531, 23.362, 20.671, 24.244, 37.948, 14.597 and 5.170, all P<0.05). The familial H. pylori infection rate was 87.59% (233/266), and in 61 families all member were infected (26.18%, 61/233). The positive rate was 23.08% (6/26) in 50 families with children under 18 years when both parents were infected. Among 231 coupled families, both couples were infected in 78 families (33.76%), one couple was infected in 113 families (48.92%), and both couples were not infected in 40 (17.32%). With the increase of marriage time, the infection rate of both spouses increased significantly ( χ2=7.775, 12.662, 15.487, all P<0.05). Conclusions:The distribution of H. pylori infection presents a family cluster pattern, and intrafamilial infection is an important transmission rout of H. pylori. The type I strain of H. pylori is the dominate strain in this area.
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Objective:To investigate the mechanism of cathepsin G(CatG) in improving the treatment efficacy of T cells in gliomas.Methods:(1) Clinical data of 397 glioma patients in the glioma database were collected, Kaplan-Meier method was used to perform survival analysis, and the correlation between CTSG and β2-microglobulin ( β2M) mRNA expressions in glioma tissues was analyzed. (2) Glioma stem cell (GSC) 387 and GSC3565 were isolated from glioblastoma and differentiated into differentiated glioma cell (DGC) 387 and DGC3565, respectively; GSC387 was divided into CatG group and CatG inhibitor group, and cells in the CatG group and CatG inhibitor group were cultured with 0.1 μg/μL recombinant human leukocyte antigen (HLA)-A*02:01 and HLA-B*15:01 combined with 4 ng/μL CatG or 10 mol/L CatG inhibitor for 10 min, respectively; the expressions of HLA-A*02:01 and HLA-B*15:01 were detected by Thomas bright blue staining, and the protein expressions of major histocompatibility complex (MHC)-I and MHC-DR were detected by Western blotting. (3) GSC387, GSC3565, DGC387, and DGC3565 were divided into 4 groups, including CatG group, CatG inhibitory group, blank antibody group 1 and blank antibody group 2, respectively; 4 ng/μL CatG, 10 μmol/L CatG inhibitor, blank antibody 1 and blank antibody 2 were added into the cells from the 4 groups for 24 h, and the expression of HLA-ABC was detected by flow cytometry. (4) GSC387, GSC3565, DGC387, and DGC3565 were divided into CatG group and CatG inhibitory group, respectively; luciferase assay was used to detect the influence of CatG in the killing effects of T cells and natural killer cells. Results:(1) The survival rate in patients from CTSG mRNA high expression group was significantly higher than that in patients from CTSG mRNA low expression group, and the survival rate in patients from β2M mRNA low expression group was statistically higher than that in patients from β2M mRNA high expression group ( P<0.05); a negative correlation between CTSG mRNA and β2M mRNA expressions was noted in glioma tissues ( r=-9.160, P=0.000). (2) Thomas bright blue staining showed that the expressions of HLA-A*02:01 and HLA-B*15:01 obviously increased in the CatG group as compared with those in the CatG inhibitor group; Western blotting showed that as compared with the CatG inhibitor group, the CatG group had increased MHC-I expression, and decreased expressions of α and β chains of MHC-DR. (3) Flow cytometry showed that the HLA-ABC expressions in GSC387 and GSC3565 of the CatG group were statistically higher than those in the CatG inhibitor group ( P<0.05). (4) Luciferase assay showed that, as compared with the CatG inhibitor group, the CatG group had statistically higher proportion of T cells killing GSCs ( P<0.05). Conclusion:CatG can improve the immunotherapy efficacy in GSCs, mainly by increasing the MHC-I expression on the cell surface.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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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.
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Objective To detect the effect of plumbagin on proliferation,apoptosis and invasion ofglioma cell lines,and investigate the underlying mechanism.Methods Glioma cell lines SWO-38 and U251 were routinely cultured in vitro,and treated with various concentrations of plumbagin (0-50 μmol/L) for 48 h; cell viability changes were detected by MTT assay,and median inhibitory concentration (IC50) was calculated; after 0,2.5,5 and 10 μmol/L plumbagin treatment for 48 h,cell apoptosis was detected by annexin V/propidium iodide double-labeled flow cytometry; scratch test was used to observe the cell invasion and migration; Western blotting was used to assess the SOX2 protein expression; MiR200b,miR200c,miR-203 and miR-21 expression changes were examined by real-time quantitative PCR.Results Plumbagin dose-dependently inhibited the proliferation of the glioma cells;the IC50 values ofplumbagin in SWO-38 and U251 cells were 6.8 and 7.4 μmol/L,respectively.After 0,2.5,5 and 10 μmol/L plumbagin treatment for 48 h,cell apoptosis ratio was gradually increased,and MiR200b,miR200c,miR-203 expressions gradually increased,with significant differences (P＜0.05).Cell invasion and migration in these two cell lines were decreased and the SOX2 protein expression was decreased.Conclusion Plumbagin inhibits cell growth,induces apoptosis,and decrease cell invasion and migration,which might be related to the increase ofmiR200b,miR200c and miR-203 expressions and the decrease of SOX2 protein expression.
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Objective To analyze the relation between SOX2 and miR-200b and their influences in clinical pathology and prognosis of gliomas patients.Methods One hundred and twenty-three human glioma specimens,collected in our hospital from January 2001 to December 2005 and conformed by pathology,were chosen in our study; other 23 healthy brain tissues collected during intracranial decompression were used as controls.Real-time quantitative PCR (qRT-PCR) was employed to detect the miR200b expression; immunohistochemistry was used to detect the SOX2 expression.Relations between SOX2 and miR-200b in different grades of gliomas were analyzed; the correlations of clinical parameters with SOX2 and miR-200b expressions were analyzed; Cox proportional hazards regression model was used to analyze the influences of SOX2 and miR-200b expressions in survival times of the patients,and the survival curves of patients with different SOX2 and miR-200b expressions were compared.Results The higher the pathology grade,the lower the miR-200b expression; gliomas of grade Ⅲ and Ⅳ had SOX2 positive expression enjoyed significantly lower miR-200b expression than those had SOX2 negative expression (P＜0.05).The SOX2 and miR-200b expressions in gliomas of different pathology grades were significantly different (P＜0.05).Cox proportional hazards regression model indicated that miR-200b and SOX2 were the independent risk factors for prognosis of gliomas.In patients of grade Ⅲ and grade Ⅳ gliomas,significantly higher 5 years survival rate in patients with high miR200b expression was noted as compared with that with low miR200b expression (P＜0.05),and significantly higher 5 years survival rate in patients with negative SOX2 expression was noted as compared with those with positive SOX2 expression (P＜0.05).Conclusion The miR-200b and SOX2 expressions show difference only in poor differentiated tissues of grade Ⅲ and Ⅳ gliomas; the two have influence in the survival time.
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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.