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BACKGROUND: Recent studies have shown that Taohong Siwu Decoction can alleviate the damage of vascular endothelial cells and maintain their normal secretory function, and endothelial progenitor cells can promote the repair of endothelial injury. Therefore, it is assumed that Taohong Siwu Decoction may protect endothelial function by improving the functional activity and increasing the number of endothelial progenitor cells. OBJECTIVE: To investigate whether Taohong Siwu Decoction can augment the number and functional activity of peripheral blood endothelial progenitor cells. METHODS: Endothelial progenitor cells were isolated from the peripheral blood of healthy subjects, and divided into control, low-, moderateand high-concentration Taohong Siwu Decoction groups. Cells were then cultured to observe the dose-effect relationship within 24 hours. Meanwhile, the high-concentration Taohong Siwu Decoction group was cultured for respective time points (6, 12, 24 and 48 hours) for observing the time-effect relationship. The number of endothelial progenitor cells was counted under inverted phase contrast microscope. Proliferation, adhesion and migration of endothelial progenitor cells were detected by MTT chromatometry, adhesion activity assay and modified Boyden chamber assay, respectively. RESULTS AND CONCLUSION: (1) The proliferation, adhesion and migration abilities of endothelial progenitor cells in the Taohong Siwu Decoction groups were significantly higher than those in the control group and showed a certain dose-effect relationship. (2) The proliferation, adhesion and migration abilities of endothelial progenitor cells in the Taohong Siwu Decoction groups were enhanced in a time-dependent manner, especially at 24 hours after intervention (P < 0.01). To conclude, the Taohong Siwu Decoction can increase the number of endothelial progenitor cells and promote cell functions. High-concentration Taohong Siwu Decoction exhibits the best interventional effect at 24 hours after intervention.
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BACKGROUND: Recent studies have shown that Taohong Siwu Decoction can alleviate the damage of vascular endothelial cells and maintain their normal secretory function, and endothelial progenitor cells can promote the repair of endothelial injury. Therefore, it is assumed that Taohong Siwu Decoction may protect endothelial function by improving the functional activity and increasing the number of endothelial progenitor cells. OBJECTIVE: To investigate whether Taohong Siwu Decoction can augment the number and functional activity of peripheral blood endothelial progenitor cells. METHODS: Endothelial progenitor cells were isolated from the peripheral blood of healthy subjects, and divided into control, low-, moderateand high-concentration Taohong Siwu Decoction groups. Cells were then cultured to observe the dose-effect relationship within 24 hours. Meanwhile, the high-concentration Taohong Siwu Decoction group was cultured for respective time points (6, 12, 24 and 48 hours) for observing the time-effect relationship. The number of endothelial progenitor cells was counted under inverted phase contrast microscope. Proliferation, adhesion and migration of endothelial progenitor cells were detected by MTT chromatometry, adhesion activity assay and modified Boyden chamber assay, respectively. RESULTS AND CONCLUSION: (1) The proliferation, adhesion and migration abilities of endothelial progenitor cells in the Taohong Siwu Decoction groups were significantly higher than those in the control group and showed a certain dose-effect relationship. (2) The proliferation, adhesion and migration abilities of endothelial progenitor cells in the Taohong Siwu Decoction groups were enhanced in a time-dependent manner, especially at 24 hours after intervention (P < 0.01). To conclude, the Taohong Siwu Decoction can increase the number of endothelial progenitor cells and promote cell functions. High-concentration Taohong Siwu Decoction exhibits the best interventional effect at 24 hours after intervention.
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Objective To investigate the protective effects of N-acetyl-L-tryptophan (L-NAT) on intestinal damage after rat hepatic ischemia reperfusion.Methods Twenty-four healthy adult rats were divided into the sham operation group (Sham),ischemia reperfusion group (IR),ischemia reperfusion and N-acetyl-L-tryptophan group(IR+L-NAT).The hepatic ischemia reperfusion model was established by occluding the afferent vessels of the left and middle lobes.The morphological structures of the small intestine were observed by hematoxylin-eosin (HE) staining.The expressions of active caspase-3,Bax and Bcl-2 were detected by immunohistochemistry staining.Results (1) In the IR group,the structure of intestinal villis was destroyed,the intestinal mucosa showed congestion and exfoliation,the epithelial cells had degeneration and necrosis,and infiltration of inflammatory cells appeared;which could be alleviated by L-NAT.(2)The immunohistochemistry showed that compared with the Sham group,the expression of active caspase-3,Bcl-2 and Bax in the IR group was increased,after L-NAT intervention,the Bax and caspase-3 expression was decreased,while the Bcl-2 expression was further increased.Conclusion L-NAT could inhibit the apoptosis of small intestinal epithelial cells caused by liver ischemic reperfusion and attenuates intestinal epithelial damage.
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Objective To investigate the protective effects of N-acetyl-L-tryptophan (L-NAT) on intestinal damage after rat hepatic ischemia reperfusion.Methods Twenty-four healthy adult rats were divided into the sham operation group (Sham),ischemia reperfusion group (IR),ischemia reperfusion and N-acetyl-L-tryptophan group(IR+L-NAT).The hepatic ischemia reperfusion model was established by occluding the afferent vessels of the left and middle lobes.The morphological structures of the small intestine were observed by hematoxylin-eosin (HE) staining.The expressions of active caspase-3,Bax and Bcl-2 were detected by immunohistochemistry staining.Results (1) In the IR group,the structure of intestinal villis was destroyed,the intestinal mucosa showed congestion and exfoliation,the epithelial cells had degeneration and necrosis,and infiltration of inflammatory cells appeared;which could be alleviated by L-NAT.(2)The immunohistochemistry showed that compared with the Sham group,the expression of active caspase-3,Bcl-2 and Bax in the IR group was increased,after L-NAT intervention,the Bax and caspase-3 expression was decreased,while the Bcl-2 expression was further increased.Conclusion L-NAT could inhibit the apoptosis of small intestinal epithelial cells caused by liver ischemic reperfusion and attenuates intestinal epithelial damage.
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BACKGROUND: There is an intimate temporal and spatial relationship between growth of primitive cardiac cells, septum transversum mesenchyme and liver development. The signal from primitive cardiac cells and septum transversum mesenchyme induces the ventral foregut endoderm cells specialize toward hepatocytes. While the septum transversum mesenchyme provides a suitable environment for forming the liver bud and promoting the growth and differentiation. However, the molecular mechanism of this induction is not yet delineated.OBJECTIVE: Using alpha-fetal protein (AFP), c-Met and cytokeratin (CK) 19 as markers of hepatic stem cells, the growth of early human embryo of 3-5 weeks and morphologic characteristic of hepatic stem cells were observed to demonstrate the characteristic and factors that affected the proliferation and differentiation of hepatic stem cell, which provided experimental evidence for basic research and clinical application of hepatic stem cells.DESIGN: An opening experiment was designed.SETTING: Department of Anatomy, Weifang Medical College.MATERIALS: The experiment was carried out at the Scientific Research Center of Chengdu Medical College between September 2004 and January 2005. Twenty cases fresh human embryos aged less than 2 months were collected with signed agreements of the pregnant women suffering from pregnancy termination with mifepristone. The samples were fixed with 40 g/L polymerisatum in 20 minutes and embedded routinely in paraffin, and then 5 μm thick series sections were continuously made. After hematoxylin-eosin staining, the embryonicage was determined under the microscope according to the length of embryos, the number of somites and the development of organs, which was referring to the Jirasek's staging standard of human embryo.METHODS: The immunohistochemical staining was conducted with SABC method on one of every ten sections, which were incubated overnight at 4 ℃ with polyclonal antibodies against hepatocyte growth factor (HGF),c-Met, insulin-like growth factor (IGF-Ⅰ), IGF-Ⅰ receptor (IGF-IR), transforming growth factor (TGFβ1), TGFβR1, TGFβR2 or monoclonal antibodies against proliferating cell nuclear antigen (PCNA), AFP and CK19.The following day, the sections were incubated for 2 hours at room temperature with biotinylated anti-mouse or anti-rabbit IgG and SABC liquid respectively, and then diaminobenzidine (DAB) was used to color them. The negative control was conducted with the phosphate buffer, then the sections were observed and photographed under light microscope.MAIN OUTCOME MEASUERS: ①the morphologic characteristic of human hepatic stem cells and immunohistochemical staining of markers②the expression of HGF, IGF-Ⅰ, TGFβ1 and their receptors on human embryonic livers of 3-5 weeks, primitive cardiac cells and septum transversum mesenchyme.RESULTS: ①The morphologic characteristic of human hepatic stem cells and immunohistochemical staining of markers: The hepatic bud formed at the end of 3rd week and migrated into the septum transversum mesenchyme to form the hepatic cords at the 4th week. The cells structuring the hepatic cords displayed the typical characteristic of immature cells. At the 5th week, the number of cells within the hepatic cords, the size of cell body,the cytoplasmic acidophilia all increased, whereas the basophilia of nuclei decreased. However the cellular forms were still homogeneous and displayed the typical characteristic of immature cells. The cells of hepatic cords were negative for PCNA response during 3rd-4th week but began to express positive at the 5th week, mainly in the nucleus and minority cellular cytoplasm showed weak positive. Most hepatic cells during 3rd-5th weeks were positive for AFP, c-Met and negative for CK19. The immunologic reaction depositors of AFP positive cells were located in the nuclei, cytoplasm and membrane of the hepatocytes, and c-Met presented mainly in the nuclei and the positive signal was weak in the cytoplasm. ②Expressions of HGF, IGF-Ⅰ, TGFβ1 and their receptors in the embryonic human liver, primitive heart and septum transversum mesenchyme: At the 4th week,the c-Met expressed only in all hepatocytes, whereas the other growth factors and their receptors were undetectable. At the 5th week, all the growth factors, except HGF, were expressed in the hepatocytes. The immunologic reaction depositors of TGFβ1, TGFβ1R1 and TGFβ1R2 were located in the cytoplasm and cell membrane. The positive response of IGF-Ⅰ and IGF-IR were present at nuclei, cytoplasm and cell membrane. At the 3rd-5th week, myocardial cells surrounding liver bud or hepatic cord and the septum transversum mesenchyme were positive for HGF, TGFβ1 and IGF-Ⅰ,with the signals were aggregated mainly in cytoplasm and minority nucei.CONCLUSION: ①It was at the end of 3rd week that part of endoderm cells in foregut ventral were specialized to hepatic stem cells. ②The undifferentiated hepatic stem cells could be drawn to develop to the liver stem cells with bi-directional differentiation potentials by using specific markers for studying human embryonic liver stem cells. According to the corresponding relation of embryonic age between human and rats, the time studying the rat hepatic stem cells could be calculated. ③HGF, IGF-Ⅰ,TGFβ1 and their receptors promoted the early development of human embryonic liver.
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BACKGROUND: Vascular endothelial growth factor family and its receptor play an important role in the process of angiogenesis and neovascularization. Recently, the effect of vascular endothelial growth factor family on blood has been paid much attention. OBJECTIVE: To observe the expression of vascular endothelial growth factor A (VEGFA), vascular endothelial growth factor C (VEGFC) and their receptors as well as angiopoietin-1 (ang-1),angiopoietin-2(ang-2) and their receptors in the blood island of yolk sac and aorta-gonadalmesonephros (AGM) region at embryonic 3 to 12 weeks, so as to investigate the effect of VEGF and angiopoietin family in the process of haematogenesis. DESIGN: Single sample observation. SETTING: Weifang Medical College, Experimental Center of Morphology and Staff Room of Anatomy. MATERIALS: This experiment was carried out at Weifang Medical College from February 2002 to August 2004. Specimens were collected from 47 pregnant women's 3-to-12-week abortive embryos. Informed consent was obtained from the pregnant women. METHODS: Specimens were performed successive sections. Two sections were drawn out every other 10 sections. Hematoxylin-eosin staining and immunohistochemical staining were performed. Polyclonal anti-vascular endothelial growth factor C, fms like protein tyrosine kinase(PTK), flt-4,ang-1, ang-2,Tie-2 antibody, monoclonal anti-vascular endothelial growth factor A and fetal liver kinase 1 (flk1) antibody were used for incubation overnight at 4 ℃ .Goat anti-mouse IgG and SABC solution were used separately for 2 hours. 3,3'2 diaminoazobenzidine (DAB) was used to develop. Serum of normal rabbit or mouse were used separately to replace primary antibody as negative control. They were observed and taken photographs under optical microscope. MAIN OUTCOME MEASURES: ①Blank expression of VEGFA and VEGFC and their receptors in the blood island of yolk sac and AGM region at embryonic 3 to 12 weeks. ②Blank expression of ang-1 and ang-2 and their receptors in the blood island of yolk sac and AGM region at embryonic 3 to 12 weeks. RESULTS: ①On the 21st to 25th day , vascular endothelial cells and hematopoietic cells in blood island of yolk sac strongly expressed VEGFA and its receptor fms like PTK and flk1, VEGFC and its receptor flk4, ang2 and its receptor Tie-2 protein, weakly expressed ang-1 protein. ②From the fourth week of development, dorsal aorta and mesonephros expressed above-mentioned various factors and their receptors . Immune positive reactants gathered in the large and round hematopoietic cells with nucleus. The cell quantity reached the peak at week 7. After week 8, the number of positive cells was significantly decreased, and almost all the blood cells were immune negative reaction at week 12. ③Gonad mainly expressed VEGFA , fms like PTK, flt-4,ang-1 ,ang-2 and Tie-2 protein at weeks 6 to 8 , but did not express VEGFC and flkl. ④ The expression of above-mentioned factors were not detected in the vascular endothelial cells in AGM region. CONCLUSION: Hematopoietic cells and endothelial cells of blood island of yolk sac of human embryo as well as dorsal aorta and mesonephros coexpresses various factors related to angiogenesis and haematogenesis. Haematogenesis of human embryos occurs at the fourth week.
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BACKGROUND: Femoral head necrosis can be induced in adult rabbits when a large dose of steroid has been used for a long time. However, the pathogenesis of steroid-induced femoral head necrosis needs further study.OBJECTIVE: To probe into the mechanism of the disease by light microscope and transmission microscope from morphological perspective based on the model of femoral head necrosis in rabbits.DESIGN: A randomized controlled observation.SETTING: Laboratory of Morphology; Teaching and Research Division of Pathology; Laboratory of Surgery, Weifang Medical College.MATERIALS: The experiment was carried out at the Experimental Center of Morphology, Weifang Medical College, between March 2002 and March 2003. Totally 40 adult New Zealand white rabbits were randomly divided into control group (n=10), dexamethasone group (n=10) and horse serum group (n=20).METHODS: Control group was given intravenous injection of normal saline of 10 mL/(kg·d) for 7 consecutive days. Dexamethasone group was given intramuscular injection of dexamethasone of 10 mL/(kg ·d)for 7consecutive days. Horse serum group was given intravenous administration of horse serum of 10 mL/kg; 3 weeks later the same volume of horse serum was injected once again, followed intramuscular injection of dexamethasone of 10 mL/(kg·d)for 7 consecutive days. Inferior sections of cartilage of the femoral head necrosis in the experimental animals were obtained 5 and 10weeks later, and then histological and ultrastructural changes were observed under the light microscope and transmission microscope.MAIN OUTCOME MEASURES: ① Histo-morphological observation of the animals in each group. ② Ultrastructural changes.RESULTS: All the experimental animals survived and entered the result analysis. ① Histo-morphological observation: The cells of inferior sections of cartilage of the femoral head necrosis of the experimental animals in control group were arranged regularly and had a small volume of elliptical bone cells. The cell body was located at bone lacuna, blood vessel arranged well in the medullary cavity of bone. Lesion haracteristics of femoral head in dexamethasone group and horse serum group were similar:Hematopoietic adipose in the medullary cavity of bone was significantly decreased while fat adipose obviously increased; bone trabecula of metaphysis and the inferior sections of cartilage of femoral head were found with ered, and so was the bone nucleus. The number of lacuna of bone was increased. ② Ultrastructural changes: Normal bone cells in control group were elliptical, located at bone lacuna. Nucleus was at one end of the cell with complete karyotheca and many mitochondria in the cytoplasm. In dexamethasone group and horse serum group there were lipid droplets in the osteocytes, narrowed blood capillary in the medullary cavity of bone and injured vascular endothelial cells.CONCLUSION: Corticotropin can induce necrosis of femoral head; the hormone causes accumulated fat adipose in the medullary cavity of bone.The increased internal pressure in the medullary cavity leads to ischemia of femoral head, thus inducing the necrosis of osteocytes.
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Objective:To study the effects of 17-? estradiol on capillary ultrastructure of medulla oblongata in ovariectomized rats. Methods: The 30 adult femal rats were randomly divided into ovariectomy group(A group), estradiol group(B group) and sham- ovariectomy group(C group).The rats of A group and B group were bilaterally ovariectomized ,Which were injected with normalsaline 0.1 ml/d), 17-? estradiol(20 ?g/kg?d -1) ,The rats of D group were sham- ovariectomy ,animals were injected with normalsaline(0.1 ml/d), for 6 weeks. Results:(1)The level of serum estradiol of A group was significantly decreased compared with C group(P