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Objective:To investigate the expression of CXCL1 2 and CXCR4 in adenoid cystic carcinoma(ACC)and to explore its re-lationship with clinicopathologic characteristics and prognosis of the patients.Methods:The expression of CXCL1 2 and CXCR4 in af-fected tissue was detected immunohistochemically in 62 cases of ACC.Both of the two factors and clinicalpathology factors were pro-cessed in accordance with the Kaplan-Meier method and the COX regression model.Results:The positive rates of CXCL1 2 and CX-CR4 expression were 54.8%(34/62)and 77.42%(48/62)respectively.Patients with the 2 factor expression had a shorter survival time than those without them(P<0.05).Multivariate analysis revealed that CXCR4 expression,clinical stage,histological differentia-tion and metastasis/recurrence were independent risk factors for the prognosis of ACC patients.Conclusion:The expression of CXCR4 may be correlated with the malignancy of ACC.CXCR4 expression,clinical stage,metastasis/recurrence and histological differentia-tion can indicate the prognosis of ACC patients.
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Objective:This study aimed to analyze the correlation of the expression of CXCR4, CD44, and CD133 proteins with the clinicopathological characteristics of patients to identify the factors affecting the post-operation survival rate of tongue squamous cell carcinomas (TSCCs). Methods:Clinical data of 44 patients with TSCCs were collected and retrospectively analyzed. The diagno-ses of all cases were pathologically confirmed. CXCR4, CD44, and CD133 expression in 44 TSCCs patients with different pathological grades was examined immunohistochemically. Survival curves were processed in accordance with the Kaplan-Meier method. The Cox regression model was used for the multivariate analysis of relevant clinical and survival data. Results:Among the 44 examined TSCCs patients, 29 cases were well differentiated and 15 were moderately or poor differentiated;11 cases were stageⅠ, 12 were stageⅡ, 8 were stageⅢ, and 13 were stageⅣ. Positive staining of CXCR4, CD44, and CD133 was found in all cases with different degrees. Ac-cording to the pathological tumor grade, the positive rates of CXCR4, CD44, and CD133 expression were 79.54% (35/44 cases),77.27%(34/44 cases), and 75.00%(33/44 cases), respectively. Expression of CXCR4, CD44, and CD133 significantly differed between different histological grades (P<0.05). Correlation analysis indicated that the expression of CXCR4, CD44, and CD133 was positively correlated with the metastasis, recurrence of TSCCs. COX multivariate analysis indicated that CXCR4 expression, clinical stage, and neck metastasis were independent prognostic predictors of TSCCs patients and risk factors of death. Conclusion:CXCR4, CD44, and CD133 may be correlated with the malignancy of TSCCs. CXCR4 expression, clinical stage, cervical lymph node metastasis were the correlated prognosis factors of TSCC patients after operation.
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Objective To study the expression and significance of Th17 and Tc17 cells in the peripheral blood,skin and lung in a murine model of bleomycin (BLM)-induced systemic sclerosis (SSc).Methods Thirty female BALB/c mouse were randomly divided into 3 groups,including a control group ( A group),a injected with BLM 4 week without pulmonary fibrosis(PF) group( B group) and with obviously PF group(C group).Pathological changes of skin and lung were detected.The proportion of CD4+,CD8+,CD4+IL-17+(Th17),CD8+IL-17+(Tc17) cells in the peripheral blood,skin and lung of mouse was determined by flow cytometry.The mRNA expressions of RORγt,IL-17A in skin and lung of mouse were evaluated by real-time PCR.Enzyme linked immunosorbent assay(ELISA) was used to measure the levels of IL-17 in serum.Results Dermal hydroxyproline(HYP) contents and the score of PF were significantly increased in C group [ (3.07±1.26) μg/mg,4.0±1.41 ]and B group [ (2.43±0.61) μμg/mg,1.50±0.76]as compared with A group [ (1.45±0.40) μg/mg,0.60±0.70 ],and pulmonary HYP contents was obviously increased in C group than in A and B groups,all P＜0.05.Compared with the A group,the percentage of CD4+ and Th17 cells in the peripheral blood,skin and lung of B and C groups,Tc17 cells of C group was significantly increased,and CD8+ cells was significantly decreased(all P＜0.05).The ratio of Th17/CD4+CD8+ in the peripheral blood,skin and lung of B and C groups [ ( 1.41 ±0.36)％,( 1.79±0.77)％ ],[ (2.58±1.07)％,(5.23±2.34)％ ]and [ (3.50±1.20)％,(4.02±1.32) ％ ]was significantly increased compared with A group (0.71±0.25)％,(1.15±0.59)％,(0.99±0.46)％.The ratio of Tc17/CD4+CD8+ in the lung of C groups( 1.62±0.53) ％ and in the skin of B and C groups [ (1.70±0.70) ％,( 1.63±0.63 ) ％ ]was significantly increased compared with A group [ ( 1.00±0.47 ) ％,( 1.1 1 ±0.34 ) ％ ],all P＜0.05.Compared with the A group,the mRNA levels of IL-17A,RORγt in skin of B and C groups,and in lung of C group were higher and the levels of IL-17 in serum was significantly increased,all P＜0.05.Th17 cells and the levels of IL-17 in blood were positive correlation with dermal and pulmonary inflammation,fibrosis and H YP contents,all P＜0.01.The frequency of Th17 and Tc17 cells in skin and lung respectively had a positive correlation with dermal and pulmonary inflammation,the score of fibrosis,and HYP contents of skin and lung,all P＜0.01.Conclusion Th17 and Tc17 cells were significantly increased in the peripheral blood,skin and lung of a murine model of SSc,and Th17 cells is dominated.They correlated with the inflammation and fibrosis of skin and lung,and may participate in the pathogenesis of SSc through secrete IL-17.
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ObjectiveThe expression and significance of interleukin(IL)-17 in a murine model of experimental systemic sclerosis(SSc) was studied and its correlation with transforming growth factor-beta 1 (TGF-β1) was explored.Methods Thirty female BALB/c mice were randomly divided into 3 groups,including a control group, bleomycin(BLM) injection for 4 weeks group(model 1 group) and a termination injection of BLM 4 weeks group(model 2 group).The pathological changes of skin and lung were detected.The mRNA expressions of IL-17A,RORγt,TGF-β1 mRNA were evaluated by real-time PCR.Enzyme linked immunosorbent assay was used to measure the levels of IL-17 and TGF-β1 in the serum and bronchoalveolar lavage fluid(BALF).Comparisons among groups were performed by variance analysis.ResultsSkin and lung of the model groups showed evident inflammatory cell infiltration and increased deposition of collagen fibers.The score of dermal inflammation and lung fibrosis was significantly higher in the model 1 and model 2 groups (2.5±0.8,3.0±1.8), (2.4±0.8,3.1±1.2) as compared to that of the control group (0.9±0.7,0.9±1.0),(F=12.19,8.367,25.11,4.641; all P＜0.05).The amount of hydroxyproline was markedly increased in the model groups than in the control group.Compared with those of the control group,the mRNA levels of IL-17A,RORγt,TGF- 31 in the skin and lung of the model 1 group were higher.The levels of IL-17 in serum and BALF of the model 1 group was significantly increased and the levels of TGF- β1 were increased in BALF and decreased in the serum (all P＜0.05).The mRNA levels of IL-17A in skin and lung had a positive correlation with the mRNA levels of TGF- β1,score of dermal inflammation and lung fibrosis.The levels of IL-17 in serum had a positive correlation with hydroxyproline of the skin and lung.ConclusionIL-17 may participate in systemic immune-mediated inflammation and changes of skin and lung in SSc and when combined with TGF-β1 togetter will cause damage to skin and lung in SSc.
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Objective To detect the existence of vasculogenic mimicry in hepatocellular carcinoma (HCC). Methods In this study 42 patients with a total of 47 HCC nodules underwent radical resection.Histological and immunohistochemical double staining of CD31 and PAS were applied to observe the existence of vasculogenic mimicry ( VM ). Reverse tanscription PCR (RT-PCR) were applied to study the expression of VE-cadherin, EPHA2 and MMP-2 genes. Results VM was found in 16 of the 42 (38. 1% )HCC cases. The typical forms of VM in the microscope are vessel-like structure formed by tumor cells,without endothelial cells and the PAS-positive looping pattern. The existence of VM in HCC correlates to a higher Edmondson grade, higher capacity of intrahepatic disseminating and poorer tumor-free survival time (P＜ 0. 05). Comparing the difference of VE-cadherin gene, EPHA2 gene and MMP-2 gene expression between VM positive nodes and in VM negative nodes by RT-PCR method demonstrated that VE-cadherin gene, EPHA2 gene and MMP-2 gene have a more intense expression in VM positive nodes than in VM negative nodes ( P ＜ 0. 05 ). Conclusion VM exists in human hepatocellular carcinoma. VM occurred more frequently in higher malignant HCC and predicts a higher rate of tumor recurrence and poorer prognosis.
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BACKGROUND:Study shows that margarita liquid has effect on promoting histiocyte regeneration and removing scars.OBJECTIVE:To observe the effects of margarita liquid on the proliferation of fibroblasts in human skin scar tissues.DESIGN,TIME AND SETTING:A grouping contrast observational experiment was performed in the Experiment Centre of Guangxi Medical University from September to December in 2008.MATERIALS:Scar tissue samples were obtained from patients in the Department of Plastic Surgery,Guangxi Medical University.Margarita liquid was the digest of margarita purchased from Beihai Gofar Marine Biological Industry Co.,Ltd and rich in multi-amino acids,polypeptides,vitamin,mineral matters and natural enzymes.METHODS:Human skin scar cell line was established by removing non-fibroblasts through repeated primary culture and serial subcultivation of scar fibroblasts with reference to Veelken method. The 3-5 generation human skin scar fibroblasts on exponential phase of growth were made into single cell suspension by trypsin digestion which was then inoculated on plastic 96-well cell culture plate,with the density of 0.5×10~4/well as well as 100 μL cell suspension and 100 μL DMEM medium in each well. After culture for 24 hours,primary medium was discarded. The grouping of the experiment:200 μL mediums with 125.00,62.50,31.30,15.60,7.80,3.90,1.95,0.98 mg/L margarita liquid were used respectively in margarita liquid group;200 μL pure medium was added into each well in control group.MAIN OUTCOME MEASURES:MTT and TUNEL assay were used to examine the proliferation and the apoptosis of fibroblastsrespectively.RESULTS:The 50% inhibiting concentration (IC_(50)) of margarita liquid on fibroblasts was 15 mg/L. Margarita liquid at any other concentration but 0.98 mg/L was effective in inhibiting fibroblast growth in a dose-dependent way,i.e. the higher margarita liquidconcentration,the higher inhibition ratio on fibroblast growth. Fibroblasts cultured with 15 mg/L (IC_(50)50) margarita liquid had got reduced volume,lessened cytoplasm,decreased density,increased apoptosis rate and buffy colour. Fibroblasts in control group were large,rich in cytoplasm and compact. Apoptotic index was higher in the margarita liquid group than in the blank control group CONCLUSION:Margarita liquid could inhibit the proliferation of skin scar fibroblasts cultured in vitro and induce the apoptosis of them.
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Objective To evaluate the preventive and therapeutic effects of artemisinin and artesunate on hypertrophic scar in rabbit ears.Methods Full-thickness wounds to cartilage were created in New Zealand white rabbit cars to establish animal models of hypertrophic scar.Cream was prepared with artemisinin or artesunate.A total of 96 hypertrophic scars were divided into 4 groups to receive the treatment with artemisinin cream.artesunate cream.cream vehicle(vehicle control)or no treatment(blank control)28 days after wounds were created.After 28-day treatment,animals were scarified,scars were incised and examined with HE-staining and VG-staining.Hypertrophy index.numerical density of fibroblasts and area density of collagen fibers were calculated.Results Compared with vehicle and blank controls,the scars were softer and flatter,the volume of fibroblasts decreased,and collagen fibers appeared to be more regulated and sparse in artemisiIlin or artesunate cream-treated groups.The Hypertrophy index.numerical density of fibroblasts.area density of collagen fibers were(1.452±0.27),(3638.245±463.0)cells/mm2,(32.29±6.9)%in artemisinin cream-treated group,respectively,(1.445±0.24),(3585.016±638.9)cells/mm2,(34.74±8.27)%in artesunate cream-treated group,respectively.All the three parameters were significantly reduced in artemisinin and artesunate groups than in blank and vehicle control groups(all P＜0.0 1).but no significant difference was found between artemisinin and artesunate groups (P＞0.05).Conclusion Artemisinin and artestmate cream has a reliable efficacy in the prevention and treatment of hypertrophic scar in animal models.
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Objective To investigate the levels of oxidative stress in liver tissues of hepatocelluar carcinoma(HCC)patients after transcatheter arterial chemotherapy(TAC).Methods Immunohistochemistry streptavidin biotinylated peroxidase(S-P)method was used to detect the cellular levels of 8-hydroxy-2'-deoxyguanosine(8-OHdG),p53 and p21~(waf1/cip1).Eighty-nine HCC patients were divided into TAC group(39 cases)and Non-TAC group(50 cases).15 Non-HCC liver tissues served as controls.Result 8-OHdG level was higher in Non-TAC group than that in TAC group in tumor tissues (F=9.516,P＜0.05),with that being the lowest in control group(F=9.516,P＜0.01);8-OHdG levels in cancer tissues were significantly higher than that in tumor surrounding tissues in both TAC group (t=7.101,P＜0.001)and Non-TAC group(t=8.020,P＜0.001),there was no significant difference of 8-OHdG levels between para-tumor tissues and controls.The levels of 8-OHdG between tumor and its surrounding tissues in TAC group(r=0.651,P＜0.001)and non-TAC group(r=0.493,P＜0.01)was in positive correlation.The difference of p53 levels in cancer tissues in TAC group and Non-TAC group were not statistically significant and p53 was not detected in para-tumor tissues.The difference of p21~(waf1/cip1) levels among TAC group,Non-TAC group and controls was statistically significant,the levels of p21~(waf1/cip1) in normal group was the highest(F=13.459,P＜0.001),followed by that in TAC and Non-TAC group in cancer tissues(TAC vs.Non-TAC group,P＜0.01);p21~(waf1/cip1) expression in normal controls was significantly higher than that in both TAC and Non-TAC group in para-tumor tissues(F=16.613,P＜0.001).The correlation of p21 ~(waf1/cip1) levels between tumor and its surrounding tissues was significant in non-TAC group(r=0.872,P＜0.001).Conclusions Oxidative stress levels in HCC tumor tissues were higher than in para-tumor tissues and non-HCC liver tissues.Cancer cells probably survive chemotherapy by fortifying oxidative stress repair mechanism.
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ObjectiveTo explore the effect of preoperative chemotherapy on DNA repair in hepatocellular carcinoma(HCC) patients. MethodsThe expression of hOGG1 portein in HCC and the surrounding liver tissue was detected by immunohistochemistry assay. ResultsThe expression of hOGG1 protein in HCC tissue was significantly higher in patients undergoing preoperative chemotherapy than that in control cirrhotic tissues,that of paracancerous tissues,and in patients without preoperative chemotherapy( ?~2=4.8297,?~2=4.0292,all P
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Objective: To study the effect of a potent angiogenic in hi bitor TNP-470 on the growth of sarcoma. Methods:1?10 6 sarcom a S-180 cells 0.1 ml were inoculated into submandibular region in each of 40 K M mice. The mice were divided into control and treatment groups with 10 in each group. Treatment was started 8 hours after inoculation. TNP-40 at 10 mg/kg, 30 mg/kg and 100 mg/kg was given subcutaneously every other day in the 3 treatment groups,total 6 times. On the 12th day, the mice were sacrificed, tumor and mice were weighted. Apoptosis of tumor cells was observed by TUNEL method and transmi ssion microscope. Express of VEGF, bFGF were detected by immunohistochemical sta ining. Results:Sarcoma was developed in all of the mice. The sa rcoma cells invaded deep into adjacent organs and tissues such as muscle, subman dibular gland, parotid and facial nerve. 10 mg/kg, 30 mg/kg and 100 mg/kg TNP- 470 inhibited the growth of the tumor by 27.62%,63.81% and 85.71% respective ly, increased the apoptosis cell number by 54.46%,156.69% and 432.48% respect ively (P
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Objective To determine the effect of intravenous administration of vascular endothelial growth factor (VEGF) on myocyte apoptosis and the expression of P 53 , Fas, Bax and Bcl 2 in an experimental model rat with acute myocardial infarction Methods Thirty three male Sprague Dawley rats were subjected to left coronary artery ligation The rats were randomized to receive VEGF 165 heparin(treated group) or heparin saline(control group), respectively, 24 hours after surgery VEGF 165 (2 ?g/1 ml)with heparin(50 U)or heparin saline(50 U/1 ml)was administered daily via tail vein for 7 days(treated group, n=8;control group, n =10) and 14 days(treated group, n=7;control group,n =8)separately Apoptotic index and the expression of the P 53 , Fas, Bax and Bcl 2 in myocardium were measured at 9 or 16 days after coronary artery ligation Results The number of apoptotic myocytes in VEGF treated group was less than that in control group\[(10?2)% vs (28?2)%, P