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Objective To investigate the expression of nuclear factor kappaB(NF-κB) and regulated upon activation normal T cell expressed and secreted chemokine(RANTES) during the formatiom of ascending aortic aneurysm. Methods Forty Wistar rats were randomly divided into the control group(n=20) and the experimental group(n=20).The rat models were made by ligating the ascending aorta. The ascending aortas were taken after ligation for 3months. Immunohistochemistry staining was performed to detect the protein expression of NF-κB and RANTES. The expression of NF-κB and RANTES mRNA were detected by RT-PCR. Results Immunohistochemisry staining results showed NF-κB and RANTES expression significantly increased in aneurysm, while there was a little positive staining in the control group. RT-PCR results indicated that the expression levels of NF-κB and RANTES in the aneurysm were stonger than that of the control group. The expression of NF-κB and RANTES mRNA were remarkably correlated. Conclusion The expression of NF-κB and RANTES in ascendin aortic aneurysm are stronger than that in the control. NF-κB and RANTES may contribute to the pathogenesis of the ascending aortic aneurysm.
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BACKGROUND: The glycoprotein which ends of a-galactosyi residues (a-Gal) is the major heterogenic antigen for hyperacute rejection. OBJECTIVE: To observe the distribution characteristic of the small tissue engineered vessel scaffold, and a-Gal in endothelial cells and smooth muscle cells of Wistar rats and Japanese white rabbits, in addition, to discuss the feasibility of applying acellular tissue vessel scaffold to heterogeneous blood vessel transplantation. DESIGN, TIME AND SETTING: The contrast observation was conducted at the Department of Human Anatomy of China Medical University between March 2003 and December 2004. MATERIALS: Totally 30 caudal arteries of Wistar rats were collected. Fifteen of which were prepared for small vessel scaffold (small vessel scaffold group), the remained 15 served as caudal artery group. Additionally, 15 central arteries were obtained from each ear of Japanese white rabbits (central artery group). METHODS: Totally 16 mg/L Bandeiraea Simplicifolia I Isolectin B4 (BSI-B4) was added for DAB staining with Affinity histochemistry method. Then MetaMorth/C5050/BX41 microscopic image analysis system was used to detect the positive reaction product of a-Gal. MAIN OUTCOME MEASURES: Color changes of vascular wall were observed under light microscope; the optical density of the positive reaction product of a-Gal was measured. RESULTS: The expression of a-Gal was mainly located in the cell membrane, as well as cell nucleus of endothelial cells in the central artery group. The expression of a-Gal of endothelial cell was strong positive in the caudal artery group, which was weak or negative expressed in the small vessel scaffold group. The optical density of a-Gal expression was lowest in the tunica intima of small vessel scaffold group, which was less in the central artery group than the caudal artery group (P < 0.001). The a-Gal expression in the tunica media of small vessel scaffold group was less than the central artery and the caudal artery groups (P < 0.001). CONCLUSION: In the caudal artery of Wistar rat, the expression of a-Gal is higher than that in the central artery of Japanese white rabbit. Therefore, the heterogeneous tissue engineered vessel material from acellular caudal artery of Wistar rats can be used in blood vessel transplantation.
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Objective To evaluate the growing condition of human umbilical vein endothelial cells (HUVECs), which were cultured on the membrane of different component, such as Chinese medicine, before or after the alkali treatment of 3-hydroxybutyrate-co-3-hydroxyhexanoate copolyesters (PHBHHx) and the biocompatibility between PHBHHx flim and endothelial cell. Methods The HUVECs were harvested by Baudine method,and identified by immunohistochemical method.Then the HUVECs of third passage were inoculated on the material surface and cultured for 8 hours,12 hours and 24 hours. After that, the morphology of HUVECs on different surfaces were observed by scanning electron microscopy, the distribution condition on different membranes were compared by cell-labeling immunofluorescence, and the cell viabilities of all groups were detected by MTT method. Results The HUVECs were successfully separated,and immunohistochemistry staining of FLK-1 and factor Ⅷ was positive.The result of HUVECs culture showed that cells on the material surface growed well, and proliferated significantly. The MTT analysis showed that the PHBHHx film of surface modification and adding some certain proportion of Chinese medicine could promote the growth and proliferation of HUVECs in vitro,and the cells were thriving, full shape, distribution on the surface by scanning electron microscopy and fluorescence microscopy.Conclusion The PHBHHx film of surface modification and containing certain proportion of Chinese medicine coating had good compatibility of HUVECs, which was favourable to cell growth, adherence and proliferation in vitro.
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Objective The experiment aims at probing the best condition of the isolation and purification of rat islets.Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation.Then the purified islets were subjected to histological staining,electron microscopy and radioimmunoassay for identification of specificity and viability.Results The histological staining revealed that the viability and the purity of the purified islets were above 95％and 85％respectively.Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules.Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly,which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion.There aye many factors that influence the quantity and quality of the acquired islets,such as the completed expansion of pancreas,the concentration and viability of collagenase and the digested time,etc.
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<p><b>OBJECTIVE</b>To determine the feasibility and accuracy of high-frequency ultrasound in evaluating the left ventricular (LV) structure and function in normal and pressure overload rats and to examine the changes of the left ventricle during its transition from hypertrophy to heart failure.</p><p><b>METHODS</b>Thirty-eight female rats were randomly assigned to normal (n = 10), operated (n = 16) and sham-operated (n = 12) groups. Parasternal long axis and short axis images were acquired by a 7.5 mHz linear ultrasound probe at 12 weeks and 20 weeks after the operation respectively.</p><p><b>RESULTS</b>Left ventricular structure and function could be satisfactorily imaged for dimensions and mass. Compared to the sham-operated groups, at 12 weeks after the operation, the operated rats had increased LV wall thickness and mass (P < 0.01) with normal cavity and FS% (P > 0.05). At 20 weeks after the operation, the LV wall thickness showed no further progressive change and the LV mass increased greatly with slightly dilated LV cavity and decreased FS% (P < 0.05).</p><p><b>CONCLUSIONS</b>High-frequency echocardiography provides a useful means to noninvasively evaluate LV dimensions, mass and function in rats. It will have great value for evaluating LV remodeling during the transition from LV hypertrophy to heart failure, as well as the effects of intervening drugs.</p>
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Animals , Female , Rats , Disease Progression , Echocardiography , Methods , Heart Failure , Diagnostic Imaging , Pathology , Heart Ventricles , Diagnostic Imaging , Pathology , Hypertrophy, Left Ventricular , Diagnostic Imaging , Pathology , Rats, Wistar , Stress, Mechanical , Ventricular Pressure , PhysiologyABSTRACT
Objective To investigate the expressions and significance of transforming growth factor-beta 1(TGF?1) and its typeⅡ receptor(TGF?RⅡ) in experimental rat ascending aortic aneurysm of rat model.Methods The rat ascending aortic aneurysm models were made by banding ascending aorta of young Wistar rats.The ascending aortas were taken 4 months after operation.Immunohistochemistry staining and Western blotting were used to investigate the expressions of TGF?1 and TGF?RⅡ.Result Immunohistochemistry staining results showed that TGF?1 expressed in all layers of the aortic aneurysm and the control.TGF?RⅡ was extensively located in the hyperplastic intima and tunica media smooth muscle cells in the aortic aneurysm,while there was only a little positive staining in the control group.Western blotting results indicated that the expression levels of TGF?1 and TGF?RⅡ in the aortic aneurysm were stronger than the control,P
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Objective To investigate the activity changes of gelatinase in the formation of ascending aortic aneurysm.Methods Thirty five young Wistar rats were divided into two groups:the control group and the experiment group.The rat models induced by ascending aorta banding were made.The ascending aortas were taken after 3-5 months operation,changes of gelatinase activity was observed by gelatin zymography and film in situ zymography.Results Gelatinase activity of ascending aortic aneurysm was significantly increased compared with that of normal ascending aortic aorta.Conclusion Elevation of gelatinase activity may play a significant role in the formation of ascending aortic aneurysm.
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Objective To localize endogenous alkaline phesphatase in human neutrophils with ELF(enzyme-labeled fluorescence)-97 phosphate,which yields an intensely fluorescent yellow-green precipitate at the site of enzymatic activity. Methods Neutrophils were isolated from human blood and fixed,then histochemistry with the ELF-97 phosphate was performed with the ELF-97 endogenous phosphatase detection kit.All photography was performed with a Nikon labophot fluorescence/DIC microscope. Results Fluorescent yellow-green granules,well-distributed but size-varied,were observed in neutrophils under fluorescence microscope.There were 94.102?3.133 percent rate of positive neutrophil alkaline phosphatase among 51 cases of healthy volunteers.Conclusion ELF-97 endogenous phosphatase detection kit can provide sensitive,high-resolution localization of endogenous phosphatase activity in neutrophils.
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Objective To examine the chracteristics of distribution of the FGF R before and after the flap transplantation and the influence of exogenic bFGF on the distribution of FGF R. Method Caudally based random skin flaps were raised on the backs of Wistar rats,bFGF was instilled under flaps after closure.An immunohistochemical techniques stanining method was used. Results FGF receptors distributes in the stratum basal of epidermis,the vascular endothelial cells in the dermis and hypodermis,the fibroblasts and the epithelial cells of the hair follicle and the sweat gland.In compared with preoperation,the quantity of increased gradually from the 1st day to the fifth day after operation,and the peak value is on the fifth day,it then recovered to the pre operation level on the serventh day.After applying the exogenic bFGF,the positive immunoreactive cells in the experimental group,increased remarkably from the Ist day to the 3rd day after operation in contrast with the control group.The peak value is on the 3rd day which was two days earlier than that of the control group and the peak value of the experimental group is remarkably higher than that of the control group.Conclusion\ FGF R distributes in the stratum basal of epidermis,the vascular endothelial cells in the dermis and hypodermis,the fibroblasts,and the epithelial cells of the hair follicle and the sweat gland.In compaired with pre operation,the quantity of the positive immunoreactive cells of FGF R were increased gradually from the Ist day to fifth day after operation,and the peak value is on the fifth day.After applying the exogenic bFGF,the positive immunoreactive cells of FGF receptors were increased.\;[
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Objective: Our purpose was to investigate the changes of adrenocorticotropin (ACTH), Cortisol (Cs) and prolactin (PRL) levels in peripheral blood of patients with active systemie lupus erythematosus(SLE). Methods： Radioimmunoassy. Results： Active SLE patients had a low level of ACTH,Cs in peak phase and a high level of PRL. Consequentaly, the ratio of PRL/Cs increased, which was positively correlative to active index of SLE. Conclusion： Endogenetic secretion in sufficiency of glucocorticalsteroid and increase of PRL have important function in the initiation and development of SLE.
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Objective:Our purpose was to establish an ideal chronic pressure-afterload heart failure rat model which has the transition from cardiac hypertrophy to heart failure. Methods: Chronic pressure-afterload heart failure rat model was induced by gradually constricting the ascending aorta of young rats. Young rats were randomly divided into 2 groups: the constricted and sham-operated groups. Clinical manifestation, tail-cuff blood pressure, organ weight, and hemodynamic data were observed at various time after operation. Results: The overall survival rate was 87%. Tail-cuff pressure began to increase in 4 weeks after operation. Left ventricular hypertrophy appeared in 12 weeks and heart failure in 5 months. Conclusion:It's a practical and reproducible model of cardiac hypertrophy that progresses to chronic heart failure.
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Objective To verify the safety and efficacy of percutaneous coronary intervention of Type Ⅳ bifurcation lesion of left main by using a left main bifurcation strategy and crush stenting technique with domestic drug-eluting stents(DES).Methods The study population consisted of patients with isolated unprotected ostial stenosis of the left anterior descending(LAD)or circumflex(LCX)artery.Sequential steps of crush stent deployment and post-dilation were undertaken followed by a modified crush stenting technique with domestic DES.Clinical and angiographic follow up was obtained to assess the primary endpoint of death,non-fatal myocardial infarction(MI)or target lesion revascularization(TLR).Results Twenty-nine patients(21 males,8 females)with a mean age of 62.57?14.21 years were evaluated.All patients were successfully treated using crush stenting technique with which final kissing balloon inflations were performed.The radial approach was utilized in 44.8% of the procedures.The mean procedural time was 36.2?9.4 minutes while the mean fluoroscopic time was 18.3?3.5 minutes.LAD ostial lesion was found in 58.6% of the patients.Predilatation with balloon angioplasty was performed in 44.8% of the patients.Partner stents and Excel stents were used in 79.3% and 20.7% of the patients respectively.The average stent diameter was 3.76 mm and the average stent length was 18.19 mm of main branch.GP IIb/IIIa inhibitors were used in 6(20.7%)patients.Angiographic results from Quantitative coronary angiographic(QCA)data showed mean target lesion length was of 13.20?4.71 mm and the baseline ostial stenosis was 78.4%.Follow-up angiography at a mean interval of 11.5?2.7 months revealed late lumen loss of 0.06?0.10 mm and 0.21?0.12 mm in the main branch and in the side branch,respectively.Binary restenosis did not occur within the main branch and side branch stents.Clinical follow up was available in all patients with mean duration of 14.2?5.2 months.No cardiac death,non-fatal MI occurred and no TLR needed during the followup of all patients.Conclusion The application of modified crush stenting technique and final kissing balloon inflations with domestic DES may be a reasonable option for the treatment of Type Ⅳ bifurcation lesion of left main.
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Aim To observe the effects of Ginsenoside on left ventricular remodeling after pressure-overload hypertrophy in rats.Methods After stenosis of the ascending aortic artery,20 survived female Wistar rats were randomly assigned to two groups:hypertrophy control(n=10)and Ginsenoside(100 mg?kg?d-1,n=10);sham operated rats(n=10)were selected randomly as nonstenosis control.Four weeks after the operation,the LVPW,IVS and LVDD of each rat were detected by echocardiogram.Myocardial cell and interstitial tissue were observed by immunofluorescence double staining.Results Compared with those in hypertrophy group,the LVPW and IVS in Ginsenoside group were all significantly decreased(P
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Objective To understand the mechanism of how cardiomyocytes exit from the cell cycle,we examined the expression of polo-like kinase 1(plk1) in the postnatal developmental process of cardiac myocytes. Methods Mitotic Index(MI) of cardiomyocytes was examined in the neonatal,2-week-old,4-week-old,and adult rat hearts(five cases per groups) by double immunofluorescence stained with H3P and ?-sarcomeric actin antibodies.plk1 mRNA and protein expression during the postnatal developmental process of cardiac myocytes were detected by RT-PCR and Western blot analysis in rat hearts. Results The MI of cardiomyocytes in 0-day-old hearts(0.905?0.087%) was approximately 2.4 times over that in 2-week-old hearts(0.372?0.094%)(P
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Objective To investigate the morphology and the cause of the ascending aortic aneurysm induced by ascending aorta banding. Methods Forty young Wistar rats were divided into two groups:the control group (10 rats) and the experiment group (30 rats).The rat models induced by ascending aorta banding were made.The ascending aortas were taken after operation in 3-5 months,and special staining and immuohistochemical staining technique were performed and observed under light microscope. Results The ascending aortic aneurysms were induced by ascending aorta banding of the young Wistar rats 3-5 months after operation.The occurrence of the aneurysm is 63.3%,and the occurrence of dissecting aneurysms is 36.7%.The expression of MMP-2 and MMP-3 is strong in the ascending aortic aneurysm.Conclusion The occurrence of ascending aortic induced by banding ascending aorta of the young Wistar rat is high,and the expression of MMP-2 and MMP-3 is strong.
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The cell volume, length, cross-section area, surface area, width and thickness of isolated cardiac myocytes were measured by using of a new computer system of coulter channelyzer and avias eye imaging analysis system. The rats, 3 weeks to 24 weeks in age after birth, were performed thoracic operation the morphological growth characteristics of isolated cardiac myocytes were observed.1. Physiological growth of normal cardiac myocytes was due to increase in cell volume, i. e. the cell length and cross-section area were enlarged. In the mean time, the changes in cross-section shape of myocytes happened when the myocytes growing.2. The major axis of cross-section area of myocytes was enlarged but the minor cross-section diameter did not show any more changes. So that, the growth lead to the cardiac myocytes be flattened in shape. The flattened growing pattern was one of the characteristics of normal growing myocytes. It might be very significant for the clinicians to eliminate the factors which would stimulate myocytes to be enlarged in cell width, and to treat and to prevent the heart disease.
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I. Under scanning electron microscope, the vascular network of the cutaneous, subcutaneous, and deep fascia in planta can be divided into five layers according totheir vascular morphological characteristics. They are as following: the papillary layer, the subpapillary layer, the deep dermal layer, the subcutaneous layer, and the deep fascia layer. 2. The papillary layer is the most superficial one, it is easy to recognize that there are five types of capillary network: the single loop, the multiple loop, the tower-like type, the finger-like type, and irregular type. 3. The network in the subpapillary layer is polygon-like, parallel to the surface of skin. 4. In the deep dermal layer, there are many vascular balls in the vascular trunks. 5. The subcutaneous layer: The vascular networks are limited by fat lobules. 6. The vascular-trunks run across each other and anastomose each other in deep fascia layer.
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Objective To observe the possibility of inducing mesenchymal stem cells(MSCs)to differetiate into hair follicle stem cells by the supernatant of cultured hair follicle in vitro and to investigate the transdifferentiation potentiality of MSCs. Methods MSCs were isolated and cultured from rat bone marrow by complete adherence.MSCs of passage 3 were characterized with markers CD44 and CD29 by immuohistochemical staining technique.The stem cells were induced by the supernatant of cultured hair follicle.The morphological character was observed by inverse phase-contrast microscopy.The expression of keratin 15 was detected by immunohistochemical staining technique and immunofluorescence staining technique.RT-PCR was further used to detect the expression of keratin 15. Results The isolated and separated MSCs were immunostaining positive in CD44 and CD29.After induced by the hair follicle conditioned medium in vitro,MSCS could be partially identified by the positive staining for keratin 15,a specific antibody of hair follicle stem cells.After 3 weeks′ induction,keratin 15 was detected by RT-PCR in MSCs induced by supernatant of cultured hair follicle.Conclusion MSCs have the potential to differentiate into hair follicle stem cells induced by the supernatant of cultured hair follicle in vitro.
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Objective The experiment aims at probing the best condition of the isolation and purification of rat islets. Methods The islets were isolated from rat pancreas by hepatopancreatic duct perfusing with collagenase and purified with Ficoll 400 discontinuous density gradient centrifugation.Then the purified islets were subjected to histological staining,electron microscopy and radioimmunoassay for identification of specificity and viability. Results The histological staining revealed that the viability and the purity of the purified islets were above 95% and 85% respectively.Electron microscopy showed that the purified islets were morphologically intact with clear membrane and plenty of secreting granules.Radioimmunoassay demonstrated the secreted insulin concerntration between low-glucose groups and high-glucose groups varied significantly,which verified the good function of the islets.Conclusion Hepatopancreatic duct perfusing with collagenase is a good method for digestion.There are many factors that influence the quantity and quality of the acquired islets,such as the completed expansion of pancreas,the concentration and viability of collagenase and the digested time,etc.
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Objective To culture the osteoblast of Wistar rat in vitro and study its biological characteristics. Methods With heat-cool alternative method and by way of the trysin digestion, the osteoblast cells, obtained from femoral bone of Wistar rats, were colleced and cultured with special cultural fluid in vitro. After about two weeks, cells lined up in one layer. Then cells were generationed and examined by phase contrast microscope, H E strain, ALP strain, type Ⅰ and type Ⅲ collagen strains. Results The cells had the same morphological feature with ALP activity, type Ⅰ collagen strain positive. Conclusion The cultured osteoblasts derived from bone mass were seed cells for the bony tissue engineering, which is the base for the tissue-engineering born construction in vitro.