الملخص
Objective To determine the time point when porcine pancreatic elastase(PPE)induced abdominal aortic aneurysm(AAA)reaches the regression phase in mice and observe the histological characteristics of AAA in regression phase.Methods AAAs were induced by transient intraluminal infusion of PPE in C57BL/6J mice.The diameters of the mouse abdominal aortas were measured before PPE infusion and sacrifice time,day 14 for AAA progression phase or day 56 for regression phase after PPE infusion,respectively.The histological characteristics of the aneurysm lesion site on day 14 and day 56 after surgery were compared and analyzed.Results The diameters of the abdominal aortas were significantly increased in both day 14 and day 56 after PPE infusion groups(diameter growth rate 147％and 155％,respectively)as compared to the baseline diameters.In the day 14 group,the infused aortas showed typical AAA characteristics,such as elastin break/degradation,medial smooth muscle cells depletion,and inflammatory cell diffused infiltration.In the day 56 group after PPE infusion,although the artery diameter did not change significantly as compared to the day 14 group,histology showed that elastin was partially repaired,new smooth muscle cells were added to the damaged aorta media,the infiltrated inflammatory cells were significantly subsided,and the adventitia neovascularization was reduced,showing a significant feature of the disease regression phase.Conclusion In the PPE-induced mouse AAA model,day 56 after surgery is an appropriate time point for observing aneurysm regression,and the histological characteristics of the regression are obvious.
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【Objective】 To study the effect of macrophage mediator 1 (MED1) deficiency on atherosclerosis in female mice. 【Methods】 ApoE knockout (ApoE-/-), LDLR knockout (LDLR-/-), MED1fl/fl, and macrophage MED1 knockout (MED1△Mac) mice were recruited in the study. Two types of mouse model were constructed:ApoE and macrophage MED1 double knockout (MED1△Mac/ApoE-/-) mice and their littermate controls (MED1fl/fl/ApoE-/-). ② LDLR knockout (LDLR-/-) mice receiving bone marrow from MED1△Mac (MED1△Mac→LDLR-/-) or MED1fl/fl (MED1fl/fl→LDLR-/-) mice. Female mice from these two models were fed a Western diet (21% fat and 0.15% cholesterol) for 12 weeks to promote the development of atherosclerosis. Body weight, total cholesterol (TC), and total triglyceride (TG) content in plasma were measured dynamically. After Western diet feeding for 12 weeks, aortic tree and aortic root were collected and hematoxylin-eosin (H&E) and oil red O staining were performed. 【Results】 Plasma TC and TG did not significantly differ between MED1fl/fl/ApoE-/- control group and MED1△Mac/ApoE-/-experimental group. However, the plaque area in aortic tree and aortic root was significantly increased in MED1△Mac/ApoE-/-mice. Moreover, compared with that in MED1fl/fl→LDLR-/- control group, the plaque area of aortic tree and aortic root had an increasing trend in MED1△Mac→LDLR-/- mice group. 【Conclusion】 MED1 deficiency in macrophages promotes the development of atherosclerosis in female ApoE or LDLR knockout mice.
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【Objective】 To compare the histological characteristics of porcine pancreatic elastase （PPE） induced abdominal aortic aneurysms （AAA） and angiotensin Ⅱ （AngⅡ） induced AAA in mice. 【Methods】 In the PPE group， the mouse abdominal aorta segment from the infrarenal abdominal aorta to the iliac artery was isolated and its branch arteries were ligated to avoid leakage during PPE perfusion. We perfused the isolated aorta segment with a PPE solution at a concentration of 1.5 U/mL for 5 min and then closed the abdominal cavity. The diameter of the abdominal aorta was measured before and 14 days after the surgery， and the perfusion segment of the arteries was collected at day 14 after the surgery. The histological characteristics of the aneurysm were analyzed and graded by histological and immunohistochemical methods. In the AngⅡ group， ten apolipoprotein E knockout mice were prepared， and AngⅡ [1 000 ng/(kg·min)] was infused with osmotic pumps for 28 days. The aorta was separated and the aneurysm aorta segment was analyzed. The wild type mice were used as normal health controls. 【Results】 In the PPE group， the diameter of the PPE perfused aorta segments increased and was significantly larger than the basal diameter ［（0.52±0.02） mm vs. （1.23±0.11） mm］ at day 14 after surgery. All the ten mice developed AAA after PPE application. The histological results showed typical pathological features of AAA in PPE perfused mice， such as elastic fiber breakage， smooth muscle exhaustion， and increased inflammation. Six of the ten mice developed aneurysms after AngⅡ infusion （6/10）. The aneurysms/dilatations were mostly in the suprarenal abdominal aorta， but also in the thoracic aorta and aortic arch. The histology analysis showed that the formation of arterial dissection was common after AngⅡ infusion， and the typical vascular “false lumen” was found. The breakage of elastic fibers， the exhaustion of smooth muscle damage， and the inflammatory response were not as typical as the PPE model in AngⅡ perfused animals. 【Conclusion】 The histological characteristics of PPE induced AAA are very typical and well present the inflammatory process in the development of aneurysm. The AngⅡ model is suitable for the study of aneurysms combined with aortic dissection. Both models have their own advantages and can complement each other.
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Objective To investigate the expression and significance of serum thymidine kinase-1(TK1), soluble intercellular adhesion molecule-1(sICAM-1),miR-210,and carcinoembryonic antigen(CEA)in pa-tients with colorectal cancer.Methods A total of 200 patients with colorectal cancer treated in the hospital from 2015 to 2017 were enrolled as the observation group and 50 healthy subjects were enrolled as the control group.Serum levels of TK1,sICAM-1,miR-210,and CEA were measured before and after treatment,and the trend of each indicator was analyzed.To analyze the relationship between tumor site,degree of tumor differen-tiation,clinical stage,w hether it is the first patient,lymph node metastasis and miR-210 levels in patients with colorectal cancer.Results The concentrations of sICAM-1,CEA,sTK1 and miR-210 in the observation group were significantly higher than those in the control group(P<0.05).The expression of miR-210 was related to the clinical stage and lymph node metastasis(P<0.05).T he sensitivity,specificity,positive predictive value, negative predictive value and accuracy of the combined tests including serum TK 1,sICAM-1,miR-210 and CEA test were higher than those of the single test,and was also higher than the combined tests of TK1,miR-210 and CEA.The sensitivity of the four combined tests was 75.70%,the specificity was 86.00%,the positive predictive value was 82.00%,the negative predictive value was 88.00%,the accuracy was 92.40%.Conclusion The combined detection of serum TK1,sICAM-1,miR-210 and CEA may have some value in the early diag-nosis of colorectal cancer,and can improve the sensitivity and specificity of colorectal cancer diagnosis.
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<p><b>OBJECTIVE</b>To investigate the effects of PPARγ overexpression on steatosis in mouse primary hepatocytes.</p><p><b>METHODS</b>Primary hepatocytes isolated from C57BL/6J mice were infected with either Ad/LacZ or Ad/PPARγ for 48 h. Steatosis of the primary hepatocytes was checked by Oil Red O staining. The mRNA and protein expression of adipocyte-specific genes PPARγ, aP2 and CideA were analyzed by using RT Real-time PCR and Western Blot.</p><p><b>RESULTS</b>Primary hepatocytes were small and even. Hepatocyte nuclei were round with dispersed chromatin and prominent nucleoli. Accumulated lipid droplets were observed in Ad/PPARγ-infected hepatocytes, but in Ad/LacZ-infected hepatocytes. Moreover, compared with Ad/LacZ-infected hepatocytes, the mRNA expression of PPARγ, aP2, FGF21 and CideA in Ad/PPARγ-infected hepatocytes were significantly induced, the protein expression of PPARγ and its target aP2 strongly increased.</p><p><b>CONCLUSION</b>over expression of PPARγ induces adipogenic steatosis in mouse primary hepatocytes.</p>
الموضوعات
Animals , Mice , Adipocytes , Metabolism , Adipogenesis , Cells, Cultured , Fatty Liver , Metabolism , Pathology , Genetic Vectors , Hepatocytes , Metabolism , Pathology , Mice, Inbred C57BL , PPAR gamma , Metabolism , Transfectionالملخص
Objective The aim of this study was to generate human cholesteryl ester transfer protein ( CETP) transgenic rabbits and analyze their biological properties.Methods We generated human CETP transgenic rabbits by DNA microinjection, and detected the expression of human CETP by real-time PCR and Western blot assay.The activity of CETP was measured using an activity assay kit.Results Human CETP transgenic rabbits were successfully generated by DNA microinjection.Compared with wide type rabbits, the expression of human CETP was dramatically increased in the liver of the human CETP transgenic rabbits.The plasma CETP activity was also much higher in the liver of human CETP transgenic rabbits than that of control rabbits.Conclusions The model of human CETP transgenic rabbits is successfully established by DNA microinjection.It will provide a useful tool for the studies of CETP biological function and its involvement in the mechanisms of cardiovascular diseases.
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Object To investigate the effect of PPARαactivation on PPARγ-induced fatty liver in the mouse. Methods Wild type mice ( C57BL/6) aged 4 to 5 weeks were used as animal models.All mice were divided into four groups.The mice in the first group were fed with chow diet.The mice in the second group were fed with a diet containing 0.125%Wy-14,643, an agonist of PPARa, for 8 days.The mice in the third group were injected with Ad/PPARγvia tail vein for 5 day.The mice in the fourth group were firstly fed with Wy-14,643 diet for 3 days and then injected with Ad/PPARγvia tail vein for another 5 day.Mouse livers were collected and photographed.The effect of PPARαactivation on PPARγ-induced fatty liver was observed by H&E and Oil red O staining.Results Compared with the controls, wild-type mice treated with Wy-14,643 for 8 days exhibited marked hypertrophy of hepatocytes with increased cytoplasmic eosinophil-ia and proliferation of peroxisomes.The liver size was significantly increased in the wild-type mice treated with Ad/PPARγfor 5 days, and over-expression of PPARγstrongly induced hepatic steatosis.Importantly, the wild-type mice pretreated with Wy-14,643 for 3 days and then given Ad/PPARγinjection exhibited dramatically the increase of liver size, which might be due to the dual function of PPARa and PPARγ.Compared with the Ad/PPARγgroup, the Wy-14,643 pretreat-ment group showed a reduced hepatic steatosis.Conclusions Activation of PPARαby Wy-14,643 effectively improves PPARγ-stimulated hepatic steatosis, which provides a novel target for prevention and therapy of fatty liver.
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To observe the effects of resveratrol on proliferation of human hepatocellular carcinoma cell line SMMC-7721 cells and expression of matrix metalloproteinase-9 (MMP-9) in vitro.
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Objective Watanabe Heritable Hyperlipidaemic (WHHL) rabbits with low density lipoprotein receptor (LDLr) gene mutation have provided unprecedented opportunities for the study of human atherosclerosis, in order to confirm LDL receptor gene status in rabbits, we developed a simple PCR technique to detect LDL mutations in rabbits. Methods Rabbits genomic DNA were extracted from ear biopsy, and amplified by PCR to detect 12 bp deletion mutation in WHHL rabbits. PCR products were directly digested with BglⅠ, and then applied to polyacrylamide gel electrophoresis. Results PCR products from homozygous LDLr +/+ rabbits generated 2 bands of 212 and 94 bp after BglⅠ digestion, LDLr +/- rabbits generated 3 bands (294, 212, and 94 bp), LDLr -/- animals, however, generated only 1 product (294 bp). Conclusion This modified PCR method is simple and reliable.
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Objective To investigate the interventional therapy and its value in the metal foreign matter in the stomach. Methods Eight patients with metal foreign matter in the stomach was studied. All patients were male, and their age ranged from 28 to 46 years with the mean age of 32.2 years. All patients had the medical history of swallowing metal foreign matter in compulsory detoxification or imprisonment. The catheter was inserted into the stomach lead by guide wire lubricated by paraffine. Then the guide wire was withdrawn and a 2.6 m long guide wire was folded in the middle and was inserted into the sromach through the catheter. A loop was made on the guide wire, and the loop was controlled to to hitch the forigen mater, then the guide wire was drawn out slowly . Results A total of 12 metal foreign matters in the stomach in all 8 patients were taken out safely, and no comqlications occurred. Conclusion The interventional therapy for the metal foreign matter in the stomach is simply, minimal invasive, cheap, effective, and with little complication. This therapy is a clinic treatment, the patient is glad to accept, and is the ideal therapy for foreign matter in the stomach.