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ObjectiveTo investigate the anti-liver fibrosis effect and mechanism of action of Kangxian Ruangan prescription based on hepatic sinusoidal capillarization. MethodsA total of 16 male Wistar rats were divided into Kangxian Ruangan prescription group and control group, with 8 rats in each group. The rats in the Kangxian Ruangan prescription group were given crude drug 40 g/kg body weight by gavage, and those in the control group were given normal saline by gavage. Serum containing Kangxian Ruangan prescription was prepared, and then primary liver sinusoidal endothelial cells were treated with the serum containing Kangxian Ruangan prescription or the control serum. Western blot was used to measure the protein expression of the vascular endothelial markers cluster of differentiation 31 (CD31) and von Willebrand factor (vWF), and immunocytochemistry was used to measure the expression of CD31 and vWF in cells. Scanning electron microscopy was used to observe the change in the fenestrae of liver sinusoidal endothelial cells. The t-test was used for comparison of continuous data between two groups. ResultsKangxian Ruangan prescription had no marked influence on the morphology of liver sinusoidal endothelial cells. The liver sinusoidal endothelial cells in the control group had a small number of narrow fenestrae with an occluded structure, while those in the Kangxian Ruangan prescription group had an increased number of large fenestrae with a clear structure. The results of immunocytochemistry showed that compared with the control group, the Kangxian Ruangan prescription group had significant reductions in the expression of CD31 (0.069±0015 vs 0.115±0.011, t=4.257, P＜0.05) and vWF (0.092±0.009 vs 0.132±0.014, t=4.274, P＜0.05). The results of Western blot showed that compared with the control group, the Kangxian Ruangan prescription group had significant reductions in the protein expression of CD31 (0.334±0.029 vs 0.448±0.013, t=6.245, P＜0.01) and vWF (0.560±0.047 vs 0709±0.045, t=3.966, P＜0.05). ConclusionKangxian Ruangan prescription can significantly resist hepatic sinusoid capillarization and thus prevent the progression of liver fibrosis.
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Objective@#To investigate the significance of lymphatic markers in the differential diagnosis of angiokeratoma corporis diffusum (ACD) and angioma serpiginosum (AS) .@*Methods@#Totally, 9 patients with ACD and 6 with AS were enrolled from Department of Dermatology, Xijing Hospital between 2006 and 2017, and their clinical and histopathological features were retrospectively analyzed. Skin sections from all the patients were stained for CD31, D2-40 and Prox1.@*Results@#In the 9 patients with ACD, abnormal vessels were weakly positive or negative for CD31, and positive for Prox1. Endothelial cells in abnormal vessels were locally positive for D2-40 in 4 patients with ACD, but negative for D2-40 in the other 5 patients. In the 6 patients with AS, the endothelia of hyperplastic small vessels were positive for CD31, but negative for D2-40 and Prox1.@*Conclusion@#The clinical features of a few patients with ACD are similar to those of patients with AS, and lymphatic markers have definite significance in the differential diagnosis of the two diseases.
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Objective@#To explore the effects of human erythropoietin (hEPO) on healing related transforming growth factor β1 (TGF-β1)/Smad3 signal transduction pathway in acute wounds of rats.@*Methods@#Seventy-two healthy Sprague Dawley rats were divided into normal saline control group, low dose group, middle dose group, and high dose group according to the random number table, with 18 rats in each group, after round acute wounds with diameter of 2.5 cm were inflicted on the back of rats. Rats in the 4 groups had debridement routinely. Wounds of rats in normal saline control group were covered by gauzes infiltrated with 1 mL normal saline, while wounds of rats in low dose group, middle dose group, and high dose group were respectively covered by gauze infiltrated with 1 mL hEPO in doses of 50, 100, and 150 U every day, and then the wounds were bandaged with 6 layers of dry gauze. Dressing change was performed once every day. On treatment day (TD) 3, 7, and 14, 6 rats from each group were taken for general observation and calculation of wound healing rate. Then the wound tissue samples were harvested after the rats were sacrificed for observation of expressions of CD31 and transforming growth factor β1 (TGF-β1) with immunohistochemical method. Protein expression of phosphorylated Smad3 of the wound tissue of 3 rats were detected by Western blotting. Data were processed with analysis of variance of factorial design, one-way analysis of variance, least-significant difference test, and Bonferroni correction.@*Results@#(1) On TD 3, obvious exudation and scab were observed in the wounds of rats in the 4 groups. On TD 7, the wounds of rats in low dose group, middle dose group, and high dose group were reduced compared with those in normal saline control group. On TD 14, all wounds of rats in the 4 groups were healed. On TD 7, the wound healing rates of rats in middle dose group and high dose group were significantly higher than the rate in normal saline control group (P<0.01). At the other time points, the wound healing rates of rats in the 4 groups were close (P>0.05). (2) CD31 mainly expressed in blood vessels. Except for those in low dose group on TD 3 and 7 (P>0.05), the expressions of CD31 in wound tissue of rats in low dose group on TD 14 and in middle dose group and high dose group on TD 3, 7, and 14 were significantly higher than those in normal saline control group (P<0.01). Except for those on TD 3 (P>0.05), the expressions of CD31 in wound tissue of rats in middle dose group and high dose group on TD 7 and 14 were significantly higher than those in low dose group (P<0.01). Except for that on TD 3 (P>0.05), the expressions of CD31 in wound tissue of rats in high dose group on TD 7 and 14 were significantly higher than those in middle dose group (P<0.01). (3) Except for that in low dose group on TD 3 (1.9±0.7, P>0.05), the expressions of TGF-β1 in wound tissue of rats in low dose group on TD 7 and 14 (3.3±1.0, 3.7±0.7), and in middle dose group and high dose group on TD 3, 7, and 14 (3.3±1.0, 3.6±1.0, 3.9±0.9, 3.4±0.7, 3.8±0.8, 4.2±0.4) were significantly higher than those in normal saline control group (1.7±0.5, 2.7±1.0, 3.0±0.9, P<0.01). Except for those on TD 7 (P>0.05), the expressions of TGF-β1 in wound tissue of rats in middle dose group and high dose group on TD 3 and 14 were significantly higher than those in low dose group (P<0.01). Except for that on TD 14 (P<0.01), the expressions of TGF-β1 in wound tissue of rats in high dose group on TD 3 and 7 were close to those in middle dose group (P>0.05). (4) Except for those in low dose group on TD 3 and 14 and in middle dose group and high dose group on TD 14 (P>0.05), the protein expressions of phosphorylated Smad3 in wound tissue of rats in the 3 groups at the other time points were significantly higher than those in normal saline control group (P<0.01). Except for those on TD 14 (P>0.05), the protein expressions of phosphorylated Smad3 in wound tissue of rats in middle dose group and high dose group on TD 3 and 7 were significantly higher than those in low dose group (P<0.01). Except for that on TD 14 (P>0.05), the protein expressions of phosphorylated Smad3 in wound tissue of rats in high dose group on TD 3 and 7 were significantly lower than those in middle dose group (P<0.01).@*Conclusions@#Exogenous hEPO can increase the expressions of CD31, TGF-β1, and phosphorylated Smad3 in acute wounds of rats, promote angiogenesis of wounds, and activate TGF-β1/Smad3 signal transduction pathway to promote wound healing.
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Objective To investigate the relationship between multidrug resistance of X-ray irradiated human nasopharyngeal squanous carcinoma cell line CNE1 and platelet endothelial cell adhesion molecule-1 (PECMA-1)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/Bcl-2 signaling pathway.Methods ①The CNE1 cells were cultured in incubator.Exponentially growing CNE1 cells were exposed to 50 Gy ionizing radiation administered in 25 fractions,2 Gy per fraction,once every two days,and the CNE1/R cells were collected.②CNE1/R cells were cultured for 24 h,and divided into three groups:the control group,experimental group one (CNE1/R cells were processed through 1 ∶ 600 PECAM-1 antibody for 24 h) and experimental group two (CNE1/R cells were processed through 10 μmol/L PI3K inhibitor LY294002 for 24 h).③The half maximal inhibitory concentration (IC50) values of the control group and two experimental groups of CNE1/R cells were detected by methyl thiazolyl tetrazolium (MTT).④The mRNA expressions of PI3K and Bcl-2 in the control group and experimental group one were measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).⑤The mRNA expressions of AKT and Bcl-2 in the control group and experimental group two were measured by semi-quantitative RT-PCR.Results ①The IC50 of the control group,experimental group one and two were (2.99 ± 0.02) μg/ml,(1.50 ± 0.03) μg/ml and (1.60 ±0.05)μg/ml,and the difference among the three groups was statiatically signficant (F =4 381.263,P =0.000).The IC50 of the two experimental groups were obviously lower than that of the control group (t =116.885,P =0.000;t =73.006,P =0.000).②The semi-quantitative values of PI3K in the control group and experimental group one were 1.66 ±0.01 and 0.90 ±0.05 (t =50.394,P =0.000),and the semi-quantitative values of Bcl-2 were 1.66 ±0.02 and 0.71 0.05 (t =183.165,P =0.000),and the differences were statistically significant.③The semi-quantitative values of AKT of control group and experimental group two were 1.53 ± 0.01 and 0.72 ± 0.01 (t =135.281,P =0.000),and the semi-quantitative values of Bel-2 were 1.66 ± 0.02 and 0.63 ± 0.02 (t =128.814,P =0.000),and the differences were statistically significant.Conclusion PECAM-1 induced multidmg resistance of human nasopharyngeal squamous cell carcinoma CNE1 cells may be related to the activity of Bcl-2 through the signal pathway of PI3K/AKT.
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Objective To investigate the expression and tissue distribution of CD31 in the process of the calf cortical with partial cancellous bone xenograft, to explore the healing mechanism and relationship between bone remodeling and revascularization, and to provide a basis for the further study of tissue engi-neered bone.Methods Thirty-six New Zealand rabbits were selected as our experimental animals.All ani-mals were implanted with the calf cortical with partial cancellous bone.Specimens were procured at 4, 8, 12 and 24 weeks after surgery, respectively.The expression and tissue distribution of CD31 were observed in the process healing with general condition, X-ray, histology and immunohistochemistry.The image analy-sis computer system was used for quantitative analysis.The extent of revascularization and remodeling was separately represented by revascularization indexes and new bone areas.The relationship was studied be-tween bone remodeling and revascularization.Results ⑴ The X-ray showed significant healing.⑵ The positive expression of CD31 was seen in the whole process of bone xenograft and was different at various stage and sites.By the 4 weeks and 8 weeks postoperative,the strong positive expression was seen in the pe-ripheral soft tissue of graft bone, the callus of joint and the periphery of cartilage nests.Much positive ex-pression was seen in the cancellous bone.There was seen some single, cord, and cluster vascular endotheli-al cells.The positive expression was seen in the enlarged haversian canals and periphery.By the 24 weeks postoperative,the positive expression was seen in the enlarged haversian canals and periphery with increasing newborn capillaries.⑶ The correlation analysis between new bone areas and revascularization showed a closely positive correlation ( r =0.984).Conclusions The positive expression of CD31 was seen in the whole process of bone xenograft and was different at various stage and sites.The positive expression of CD31 reflected the extent of revascularization and bone remodeling of bone grafts.The extent of revascularization of bone grafts and bone remodeling showed a closely positive correlation.
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BACKGROUND: Partial or complete necrosis of a skin flap is a common problem. Polydeoxyribonucleotide (PDRN) can be extracted from trout sperm and used as a tissue repair agent. The aim of this study was to investigate whether PDRN could improve the survival of random pattern skin flaps in rats. METHODS: Twenty-two male Sprague-Dawley rats were randomly divided into two groups: the PDRN treatment group (n=11) and the control group (n=11). Caudally pedicled random pattern skin flaps were elevated on their dorsal skin and resutured. The treatment group received daily intraperitoneal administration of PDRN (8 mg/kg/day), and the control group received fluid vehicle (NaCl 0.9%, 8 mg/kg/day) from day 0 to day 6. On day 7, the flap survival was evaluated and the harvested tissue surrounding the demarcation line of the necrotic area was stained with H&E, anti-rat vascular endothelial cell growth factor (VEGF) antibody, and PECAM-1/CD31 antibody. RESULTS: The average necrotic area of the flap in the PDRN group was significantly smaller when compared with that of the control group. Histologic and immunohistochemical evaluation showed that granulation thickness score and VEGF-positive staining cells were marked higher in the PDRN group than in the control group. PECAM-1/CD31-positive microvascular densities were significantly higher in the PDRN group when compared with the control group. CONCLUSIONS: This study confirms that PDRN improves the survival of random pattern skin flaps in rats. These results may represent a new therapeutic approach to enhancing flap viability and achieving faster wound repair.
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Animals , Humans , Male , Rats , Angiogenesis Modulating Agents , Platelet Endothelial Cell Adhesion Molecule-1 , Endothelial Cells , Necrosis , Polydeoxyribonucleotides , Rats, Sprague-Dawley , Skin , Spermatozoa , Surgical Flaps , Trout , Vascular Endothelial Growth FactorsABSTRACT
Objective To investigate the relationship between single nucleotide polymorphisms (SNPs),serum CD31 and HCC.Methods We analyzed three single nucleotide polymorphisms of CD31 gene Leu125Val,Asn563Ser and Gly670Arg in 190 HCC patients and 210 age and sex matched controls in a Chinese population,using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) strategy,and the serum level of CD31 was determined by enzyme-linked immunosorbent assay (ELISA).Results The distributions of CD31 gene Asn563Ser and Gly670Arg polymorphisms were not significantly different between HCC and control group (x2 =0.491,P ＞ 0.05),but the CD31 gene Leu125Val polymorphism was significantly different (x2 =10.988,P ＜ 0.05).The relative risk suffering from HCC of Val allele was 1.583 times of the Leu allele carriers(OR =1.583,95％ CI,1.197-2.093,P =0.001) ; Serum level of CD31 Val allele carriers was significantly higher than that of no carriers(x2 =10.408,P ＜ 0.05).Consistent with the results of the genotyping analyses,CD31 gene Leu125Val,Asn563Ser and Gly670Arg polymorphisms showed strong linkage disequilibrium,the Val-Ser-Arg haplotype was associated with a significantly increased risk of HCC as compared with the controls (OR =1.496,95％CI:1.095-2.046,P =0.011).Conclusions CD31 gene Leu125Val polymorphism and its Val-Ser-Arg haplotype were associated with HCC,Val allele is an important genetic susceptibility gene for HCC.CD31 Val allele carriers may subject to higher risk of HCC with enhanced CD31 expression.
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Sclerosing angiomatoid nodular transformation (SANT) of spleen is a rare inflammatory tumor-like vascular lesion composed of angiomatoid nodules in a fibrosclerotic background. We report herein on a case of SANT in the spleen with its pathologic features, and review the related literature. A 50-year-old woman presented with mild left upper quadrant discomfort and tenderness and she showed a 6 cm-sized solitary splenic mass on computed tomography. She underwent laparoscopic splenectomy. Grossly, the spleen showed a well circumscribed round-shaped solid mass with multinodular hemorrhagic surfaces. Microscopically, the mass consisted of multiple angiomatoid nodules surrounded by collagen bundles with fibroblasts and a lymphoplasma cell infiltration. Immunohistochemically, the cells of the angiomatoid nodules were positive for CD31, CD30, CD34, alpha-smooth muscle actin, and VWF-VIII, but they were negative for CD8, anaplastic lymphoma kinase protein, and D2-40. The patient has been under close follow-up without recurrence.
Subject(s)
Female , Humans , Middle Aged , Actins , Platelet Endothelial Cell Adhesion Molecule-1 , Collagen , Fibroblasts , Follow-Up Studies , Hamartoma , Lymphoma , Muscles , Phosphotransferases , Receptor Protein-Tyrosine Kinases , Recurrence , Spleen , SplenectomyABSTRACT
Objective To investigate the expression of VEGF-C,CD31 and MVD,in human ovarian epithelial carcinoma tissues,development and metastasis of epithelial ovarian carcinoma by observing their expression and analyzing their correlation.Methods 40 samples of ovarian epithelial carcinoma tissues and 20 samples normal ovary tissues were selected from 2004 to 2005 in the third affiliated hospital of Harbin Medical University,as well as 20 cases of normal ovarian tissues samples as control using immunohisrochemical staining assay,we analyzed the expression of VEGF-C,MVD detected by CD31.The relationship of these three factors with patients age tumor stage,differentiation,pathohistological type,were analyzed.Results 67.5%(27/40)had high expression of VEGF-C,15%(3/14)had low espression.The survival of patients with high VEGF-C expression was significantly worse than that of patients with low and negative expression.Conclusion VEGF-C and CD31 may play an important role in the occurrence and development of epithelial ovarian tumors,while MVD maybe associated with the ivasion and metastasis of carcinoma.The detection of VEGF-C combined with CD31 and MVD can objectively reflect the biological behavior of epithelial ovarian tumor.
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Objective To explore the changes of expression of platelet PECAM-1,P-selectin in patients with acute cerebral infarction.Methods The platelet expression levels of PECAM-1,P-selectin in 35 patients with acute cerebral infarction were serially measured with whole blood flow cytometry 24,48 hours and 5,7,14 days after the onset,and were compared with those of 30 normal controls.Results The platelet expression levels of PECAM-1 and P-selectin in patients with cerebral infarction 24 hours after the onset(78.35?10.48,7.75?3.04 respectively)were higher than those of controls(48.89?10.84,2.18?0.83 respectively,all P
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AIM: To investigate the expression of vascular endothelial growth factor receptor-3 (VEGFR-3) and CD31 in relation to metastasis in ovarian epithelial tumors. METHODS: VEGFR-3 and CD31 expression were examined by immunohistochemical methods, VEGFR-3 positive microlymphatic count (MLC) and microvessel density (MVD) were assessed by the image analysis. RESULTS: Both MLC and MVD in ovarian epithelial carcinomas were higher than those in benign tumors(MLC, P 0 05). CONCLUSIONS: VEGFR-3 and CD31 expression correlate significantly with metastasis, MLC and MVD might predict peritumoral lymphangiogenesis and angiogenesis, which may be as a biologic marker for metastasis in ovarian epithelial tumors. [
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ObjectiveTo investigate the anti-liver fibrosis effect and mechanism of action of Kangxian Ruangan prescription based on hepatic sinusoidal capillarization. MethodsA total of 16 male Wistar rats were divided into Kangxian Ruangan prescription group and control group, with 8 rats in each group. The rats in the Kangxian Ruangan prescription group were given crude drug 40 g/kg body weight by gavage, and those in the control group were given normal saline by gavage. Serum containing Kangxian Ruangan prescription was prepared, and then primary liver sinusoidal endothelial cells were treated with the serum containing Kangxian Ruangan prescription or the control serum. Western blot was used to measure the protein expression of the vascular endothelial markers cluster of differentiation 31 (CD31) and von Willebrand factor (vWF), and immunocytochemistry was used to measure the expression of CD31 and vWF in cells. Scanning electron microscopy was used to observe the change in the fenestrae of liver sinusoidal endothelial cells. The t-test was used for comparison of continuous data between two groups. ResultsKangxian Ruangan prescription had no marked influence on the morphology of liver sinusoidal endothelial cells. The liver sinusoidal endothelial cells in the control group had a small number of narrow fenestrae with an occluded structure, while those in the Kangxian Ruangan prescription group had an increased number of large fenestrae with a clear structure. The results of immunocytochemistry showed that compared with the control group, the Kangxian Ruangan prescription group had significant reductions in the expression of CD31 (0.069±0015 vs 0.115±0.011, t=4.257, P＜0.05) and vWF (0.092±0.009 vs 0.132±0.014, t=4.274, P＜0.05). The results of Western blot showed that compared with the control group, the Kangxian Ruangan prescription group had significant reductions in the protein expression of CD31 (0.334±0.029 vs 0.448±0.013, t=6.245, P＜0.01) and vWF (0.560±0.047 vs 0709±0.045, t=3.966, P＜0.05). ConclusionKangxian Ruangan prescription can significantly resist hepatic sinusoid capillarization and thus prevent the progression of liver fibrosis.