基质细胞衍生因子-1、基质金属蛋白酶-7、alpha-平滑肌肌动蛋白在人乳腺癌中的表达

目的:研究基质细胞衍生因子-1(stromal cell-derived factor-1,SDF-1)、基质金属蛋白酶-7(matrix metalloproteinases-7,MMP-7)与alpha-平滑肌肌动蛋白(alpha-smooth muscle actin,alpha-SMA)的表达。方法:选有淋巴转移的乳腺癌患者33例,在相邻切片上分别对比SDF-1及MMP-7在正常乳腺组织、乳腺癌、癌旁组织中的免疫组化的表达,同时在有SDF-1表达的邻近蜡块切片观察alpha-SMA的表达。结果:在有腋窝淋巴转移的乳腺癌及癌旁组织中SDF-1、MMP-7均强表达,与正常乳腺组织的表达比较差异有显著性,在有SDF-1表达的相邻石蜡切片存在alpha-SMA相似部位的表达;SDF-1和MMP-7表达有相关性。结论:SDF-1、MMP-7、alpha-SMA均与乳腺癌恶性行为相关,间质中SDF-1表达与肌成纤维母细胞相关;SDF-1和MMP-7存在相同联系性。

α-平滑肌肌动融合蛋白抑制成纤维细胞收缩的实验研究

目的 观察α 平滑肌肌动融合蛋白 (α SMA FP)对成纤维细胞收缩功能的影响。方法建立Wistar大鼠皮肤成纤维细胞三维立体凝胶培养模型。成纤维细胞在胶原凝胶中培养 5d后 ,实验组分别用 5、10、5 0、10 0、2 5 0mg/L的α SMA FP进行处理 ,根据α SMA FP剂量不同分为E1、E2 、E3 、E4和E5组。对照组分为空白对照 (C1)组和 2 5 0mg/Lα 骨骼肌肌动融合蛋白 (α SKA FP)处理(C2 )组。测量处理前后凝胶的直径 ,计算收缩率。检测E5组冲洗掉α SMA FP后 (wash组 )凝胶收缩率的变化情况。对传代后的成纤维细胞用α SMA FP或α SKA FP处理后进行α SMA免疫荧光染色。 结果 C1和C2 组凝胶收缩率为 (5 8.6± 3.1) %和 (5 6 .2± 4 .9) % ,二者差异无显著性意义 (P>0.0 5)。E1～E5组的收缩率分别为 (4 5 .5 6± 4 .1) %、(4 2 .3± 4 .2 ) %、(4 1.8± 3.6 ) %、(37.6±5 8) %、(2 6 .4± 4 .7) % ,wash组收缩率为 (4 8.2± 4 .9) %。E1～E5组与C1、C2 组比较 ,差异均有显著性意义 (P <0.0 5 ),与wash组比较 ,差异有非常显著性意义 (P <0.0 1)。α SMA FP着色于成纤维细胞纤维丝上 ,转染α SMA时着色不明显 ,而用α SMA FP处理后 ,α SMA着色明显。 结论 α SMA FP可特异性抑制成纤维细胞收缩 。

Heme oxygenase-1 prevents liver fibrosis in rats by regulating the expression of PPAR_γ and NF-_κB

AIM:To investigate the effects of heme oxygenase(HO)-1 on liver fibrosis and the expression of peroxisome proliferator-activated receptor gamma(PPARγ) and nuclear factor-kappa B(NF-κB) in rats.METHODS:Sixty Wistar rats were used to construct liver fibrosis models and were randomly divided into 5 groups:group A(normal,untreated),group B(model for 4 wk,untreated),group C(model for 6 wk,untreated),group D [model for 6 wk,treated with zinc protoporphyrin Ⅸ(ZnPP-Ⅸ) from week 4 to week 6],group E(model for 6 wk,treated with hemin from week 4 to week 6).Next,liver injury was assessed by measuring serum alanine aminotransferase(ALT),aspartate aminotransferase(AST) and albumin levels.The degree of hepatic fibrosis was evaluated by measuring serum hyaluronate acid(HA),type Ⅳ collagen(Ⅳ-C) and by histological examination.Hydroxyproline(Hyp) content in the liver homogenate was determined.The expres-sion levels of alpha-smooth muscle actin(α-SMA) in liver tissue were measured by real-time quantitative polymerase chain reaction(RT-PCR).The expression levels of PPARγ and NF-κB were determined by RT-PCR and Western blotting.RESULTS:The expression of HO-1 increased with the development of fibrosis.Induction of HO-1 by hemin significantly attenuated the severity of liver injury and the levels of liver fibrosis as compared with inhibition of HO-1 by ZnPP-Ⅸ.The concentrations of serum ALT,AST,HA and Ⅳ-C in group E decreased compared with group C and group D(P < 0.01).Amount of Hyp and α-SMA in the liver tissues in group E decreased compared with group C(0.62 ± 0.14 vs 0.84 ± 0.07,1.42 ± 0.17 vs 1.84 ± 0.17,respectively,P < 0.01) and group D(0.62 ± 0.14 vs 1.11 ± 0.16,1.42 ± 0.17 vs 2.56 ± 0.37,respectively,P < 0.01).The expression of PPARγ at levels of transcription and translation decreased with the development of fibrosis especially in group D;and it increased in group E compared with groups C and D(0.88 ± 0.15 vs 0.56 ± 0.19,0.88 ± 0.15 vs 0.41 ± 0.11,respectively,P < 0.01).The expression of NF-κB increased with the development of fibrosis especially in group D;and it decreased in group E compared with groups C and D(1.43 ± 0.31 vs 1.89 ± 0.29,1.43 ± 0.31 vs 2.53 ± 0.54,respectively,P < 0.01).CONCLUSION:Our data demonstrate a potential mechanism that HO-1 can prevent liver fibrosis by enhancing the expression of PPARγ and decreasing the expression of NF-κB in liver tissues.

Effects of interferon-alpha on expression of hepatic stellate cell and transforming growth factor-pi and a-smooth muscle actin in rats with hepatic fibrosis

AIM:To investigate the effect of interferon-a (IFN-α) on preventing or reversing hepatic fibrosis in rat experimental model induced by CCI4. METHODS: One hundred and ten Sprague-Dawley rats were divided into five groups: group A (normal controls, n = 18), group B (fibrotic model controls, n = 22), group C (IFN-α prevention, n = 22) initially treated with intramuscular injection of IFN-a in saline daily at the doses of 1× 105 U for 6 wk, group D (IFN-a treatment, n = 24) treated with intra-muscular injection of IFN-a in saline daily at the doses of 1×105 U for 6 wk after the first 6 wk, group E (0.9% sodium chloride treatment control, n = 24) treated with intra-muscular injection of 0.01 mL/kg daily for 6 wk after the first 6 wk. At the end of the experiment, all rats of each group were killed. Samples of the liver obtained by biopsy were subjected to histological, immunohistochemical and electron microscopic studies for the expressions of transforming growth factor-pi (TGF- μ41) and α-smooth muscle actin (α-SMA). RESULTS: The expressions of TGF-pl, the number of activated hepatic stellate cells and a-SMA in hepatic tissue of group C were significantly less than those of group B (P<0.01). The degree of fibrosis score in group B was also significantly less than that of group C under light microscope (P<0.01). CONCLUSION: IFN-a can inhibit the production of TGF-pl, decrease HSC activation and stimulate its apoptosis.

Gastrointestinal stromal tumor of duodenum:a cause of upper gastrointestinal hemorrhage

Objective:The aim of the study was to report an anemia patient with melena for five years caused by duodenal gastrointestinal stromal tumor (GIST), who required surgical treatment. Methods: A 44-year old man present with anemia appearance was admitted to our center (Department of Hepatobiliary Surgery, Union Hospital, Huazhong University of Science and Technology, China) due to sustaining melena for five years. Endoscopy found no special mucosal abnormalities in the duodenal lumen. Computed Tomography showed a well-demarcated mass, 7.4 cm in diameter, located between the C loop of duodenum and pancreatic head. Pylorus-preserving pancreaticoduodenectomy and right hemicolectomy were performed when the patient's general conditions were improved. He recuperated successfully and was discharged on the 21st postoperative day. No complications happened during the period of hospital stay. Results: Histological and immunohistochemical study revealed a high risk invasive duodenal GIST which was positive for CD117, CD34, α-smooth muscle actin and negative for S-100. Conclusion: Duodenal GIST can be a source of upper gastrointestinal hemorrhage; surgical treatment is still a reasonable choice for the patients with invasive duodenal GIST.

酸性哺乳动物几丁质酶早期诊断职业性尘肺病价值

目的观察酸性哺乳动物几丁质酶(AMCase)在矽肺模型大鼠肺组织和尘肺病患者支气管肺泡灌洗液(BALF)、血清中的表达,评价AMCase对尘肺病早期诊断的价值。方法 (1)将无特定病原体级成年雄性Wistar大鼠随机分对照组和模型组,每组15只;模型组大鼠采用动式吸入染尘法,予质量浓度为2 000.0 mg/m^(3)的游离二氧化硅粉尘每天染尘3 h,对照组大鼠不予染尘处理;2组大鼠分别于染尘第4、12和24周各处死5只。(2)采用随机数表法,选择191例进行大容量肺灌洗的职业性尘肺病患者为尘肺病组,选择12例进行大容量肺灌洗的粉尘作业人员为粉尘对照组,选择200名有粉尘作业史的健康煤矿工人为健康对照组。(3)采用蛋白免疫印迹法检测大鼠肺组织中AMCase、Ⅰ型胶原(COLⅠ)、α-平滑肌肌动蛋白(α-SMA)相对表达水平和人BALF中AMCase相对表达水平;采用酶联免疫法检测人血清AMCase水平;采用受试者工作特征(ROC)曲线评价血清AMCase蛋白水平对于尘肺病的早期诊断价值。结果模型组大鼠肺组织AMCase、COLⅠ和α-SMA蛋白相对表达水均高于对照组(P值均<0.01)。尘肺病组和尘肺病壹期、贰期、叁期亚组人群BALF中AMCase蛋白相对表达水平均高于粉尘对照组(P值均<0.05),血清AMCase蛋白水平均高于健康对照组(P值均<0.05)。ROC曲线分析结果显示:采用血清AMCase蛋白水平预测尘肺病的ROC曲线下面积为0.78(95%可信区间为0.74~0.82);当血清AMCase蛋白水平截断值为466.0 ng/L时,灵敏性为73.8%,特异性为72.6%。结论血清AMCase蛋白对于尘肺病早期诊断具有一定价值,可能是潜在的尘肺病早期诊断生物标志物。

Inhibition mechanism of compound ethanol extracts from Wuweizi(Fructus Schisandrae Chinensis) on renal interstitial fibrosis in diabetic nephropathy model mice

OBJECTIVE：To evaluate inhibition effect and mech- anism of compound ethanol extracts from Wuweizi （Fructus Schisandrae Chinensis）, Chuanxiong （Rhi- zoma Chuanxiong） and Muli （Cocha Ostreae） （FRC） on glomerular and tubular interstitial fibrosis in streptozocin （STZ）-induced diabetic nephropathy （ND） model mice. METHODS： Twenty-seven male C57BL/6 mice were divided randomly into 3 groups： nondibetic （ND）, STZ-induced diabetic （D）, and STZ-induced diabetic that were treated with .5 g. kg1. daylof FRC by oral gavage （DFRc）, with 9 in each group. The protein ex- pressions of E-cadherin, a-smooth muscle actin （a-SMA）, Plasminogen Activator Inhibitor-1 （PAl-l） in renal tissues were investigated by Western blot- ting. The expressions of fibronectin （FN） and o-SMA were detected by immunohistochemical method. The morphological changes of renal tissues were observed under a microscope. RESULTS： Renal tissues in the DFRC group showed a lessened degree of fibrosis. Meanwhile, the expres- sions of FN, o-SMA and PAl-lwere significantly lower in the DrRc group than those in the D group （all P〈0.05）.CONCLUSION： FRC can ameliorate the DN in the C57BL/6 mice, and its mechanism may relate to in- hibition on the epithelial to mesenchymal transdif- ferentiation, endothelial-myofibroblast transition and PAl-1 expression.