Effect of astragaloside Ⅳ on lipid and glucose metabolism in acute myocardial infarction through PPARγ pathway

OBJECTIVE To investigate the effects of astragaloside IV(which can be extracted from the traditional Chinese medicine Astragalus membranaceus)on lipid and glucose metabolism in acute myocardial infarction(AMI).METHODS Model of heart failure(HF)after AMI was established with ligation of left anterior descending artery on Sprague-Dawley(SD)rats.The rats were divided into three groups:sham,model and astragaloside IV treatment group.Twenty-eight days after treatment(astragaloside IV,20 mg·kg-1 daily),hematoxylin-eosin(HE)staining was applied to visualize cardiomyocyte morphological changes.High performance liquid chromatography(HPLC)was performed to assess the contents of adenosine phosphates in heart.Positron emission tomography and computed tomography(PET-CT)was conducted to evaluate the cardiac glucose metabolism.Expressions of key molecules such as peroxisome proliferatoractivated receptor γ(PPARγ),sterol carrier protein 2(SCP2)and long chain acyl CoA dehydrogenase(ACADL)were measured by Western blotting and immunohistochemistry.Oxygen-glucose deprivation-reperfusion(OGD/R)-induced H9C2 injury cardiomyocyte model was adopted for potential mechanism research in vitro.RESULTS Treatment with astragaloside Ⅳ rescued hearts from structural and functional damages as well as inflammatory infiltration.Levels of adenosine triphosphate(ATP)and energy charge(EC)in astragaloside IV group were also up-regulated compared to model group.Further results demonstrated that critical enzymes both in lipid metabolism and glucose metabolism compro mised in model group compared to sham group.Intriguingly,astragalosideⅣcould up-regulate critical enzymes including ACADL and SCP2 in lipid metabolism accompanying with promoting effect on molecules in glycolysis simultaneously.Results on upstreaming signaling pathway demonstrated that astragaloside Ⅳ could dramatically increase the expres sions of PPARγ.In vitro study suggested the efficacy of astragalosideⅣcould be blocked by T0070907,a selective PPARγ inhibitor.CONCLUSION Astragaloside IV has cardioprotective effect in improving cardiac function and energy metabolism through regulating lipid and glucose metabolism.The effects may be mediated by PPARγ pathway.

Correlation of Fibroscan parameter with serum inflammation indexes, collagen metabolism indexes and fibrosis indexes in patients with hepatitis b cirrhosis

Objective:To study the correlation of Fibroscan parameter with serum inflammation indexes, collagen metabolism indexes and fibrosis indexes in patients with hepatitis b cirrhosis. Methods:Patients who were diagnosed with hepatitis b cirrhosis in Dongfeng Hospital Affiliated to Hubei University of Medicine between June 2014 and August 2017 were selected as cirrhosis group for the research, patients who were diagnosed with chronic viral hepatitis b and without liver cirrhosis were selected as the HBV group, and healthy volunteers who received physical examination in Dongfeng Hospital Affiliated to Hubei University of Medicine during the same period were selected as the control group for the research;Fibroscan was performed to determine the liver stiffness measurement (LSM), and serum was collected to determine the inflammation indexes, collagen metabolism indexes and fibrosis indexes.Results:LSM levels as well as serum sICAM1, sP-selectin, MIF, MMP2, MMP9, CatS, TGFβ1, HA, LN, PC-III and IV-C contents of cirrhosis group and CHB group were significantly higher than those of control group whereas serum IL-21, IFN-γ, TIMP1 and TIMP2 contents were lower than those of control group;LSM level as well as serum sICAM1, sP-selectin, MIF, MMP2, MMP9, CatS, TGFβ1, HA, LN, PC-III and IV-C contents of cirrhosis group was significantly higher than those of CHB group whereas serum IL-21, IFN-γ, TIMP1 and TIMP2 contents lower than those of CHB group;LSM level in cirrhosis group was positively correlated with serum sICAM1, sP-selectin, MIF, MMP2, MMP9, CatS, TGFβ1, HA, LN, PC-III and IV-C contents, and negatively correlated with serum IL-21, IFN-γ, TIMP1 and TIMP2 contents.Conclusion: The Fibroscan parameter LSM of hepatitis b cirrhosis has evaluation value for inflammatory response, collagen metabolism and fibrosis process in the course of cirrhosis.

Correlation of the changes of TLR4/NF-κB pathway function in intrauterine adhesion tissue with the characteristics of cytokine secretion and collagen metabolism

Objective:To study the correlation of the changes of Toll-like receptor 4 (TLR4) / nuclear factorκB (NF-κB) pathway function in intrauterine adhesion (IUAs) tissue with the characteristics of cytokine secretion and collagen metabolism.Methods:The patients with IUAs who were treated in our hospital between February 2015 and March 2018 were selected as the IUAs group, and the patients who underwent hysteroscopy due to infertility and were pathologically confirmed to have normal endometrium during the same period were selected as the control group. The expression levels of TLR4/NF-κB pathway molecules and collagen metabolism genes as well as the contents of cytokines and collagen metabolism markers in the adhesion tissues of IUAs group and the normal endometrial tissue of control group were measured.Results: TLR4, NF-κB, a disintegrin and metalloproteinase 15 (ADAM15), ADAM17, matrix metalloproteinase 9 (MMP9) and plasminogen activator inhibitor 1 (PAI-1) mRNA expression as well as transforming growth factor-β1 (TGF-β1), Smad2/3, insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R), basic fibroblast growth factor (bFGF), periostin/osteoblast-specific factor 2 (Postn), type I collagen (Col-I) and actin-α (α-SMA) contents in the adhesion tissues of IUAs group were significantly higher than those of control group while urokinase-type plasminogen activator (uPA) mRNA expression was significantly lower than that of control group;TLR4 and NF-κB mRNA expression were positively correlated with TGF-β1, Smad2/3, IGF-1, IGF-1R, bFGF, Postn, Col-I,α-SMA, ADAM15, ADAM17, MMP9 and PAI-1, and negatively correlated with uPA.Conclusion:The excessive activation of TLR4/NF-κB pathway in IUAs is associated with the cytokine secretion and collagen metabolism abnormalities.

肾组织石蜡切片Ⅳ型胶原α链免疫组织化学染色在诊断Alport综合征中的应用

目的对Alport综合征(AS)患者肾脏穿刺组织进行石蜡切片Ⅳ型胶原α链免疫组织化学检测,探讨其临床应用价值。方法选取2018年11月至2022年7月浙江大学医学院附属第一医院肾脏病中心确诊的10例AS患者(AS病例组)、10例心脏死亡器官捐献者(DCD)供肾及10例确诊为免疫球蛋白A肾病患者(对照组)的肾组织进行石蜡切片Ⅳ型胶原α1、α3、α5链全自动免疫组织化学染色,将其与冰冻切片免疫荧光染色结果进行比较。将对照组肾组织石蜡切片分别采用多种方法进行抗原修复,比较修复效果。结果对照组肾组织2种染色方法Ⅳ型胶原α1、α3、α5链染色在肾小球基底膜(GBM)中均为连续线状阳性表达。AS病例组肾组织2种染色方法Ⅳ型胶原α1链染色在GBM均呈连续线状阳性表达,α3、α5链染色在GBM为阴性或节段弱阳性表达。2种染色方法染色结果具有较高的一致性(Kappa=0.615,P=0.035)。结论石蜡切片全自动免疫组织化学检测Ⅳ型胶原α1、α3和α5能较好地用于AS的诊断,为AS的诊断和研究提供了一种可靠的技术手段。

基于上皮细胞-间充质细胞转分化(EMT)理论的艾灸配合化纤Ⅳ号方对实验大鼠Collagen Type Ⅲ和PDGF干预作用实验研究

目的:基于上皮细胞-间充质细胞转分化(EMT)学说观察化纤Ⅳ号方、艾灸以及二者相配合治疗肺纤维化大鼠Collagen TypeⅢ(Ⅲ-C)和PDGF的变化,探讨其治疗效应及生物学机制。方法:将鼠龄约为6周的SD大鼠随机分为空白组、模型组、化纤Ⅳ号方组、艾灸组、化纤Ⅳ号方与艾灸配合治疗组(简称为"灸药组"),治疗30 d后处死观察其肺组织病理改变,并检测其Collagen TypeⅢ、PDGF的基因和蛋白表达情况。结果:实时荧光定量结果显示:与空白组相比,各组Ⅲ-C和PDGF m RNA表达增高(P<0.05)。与模型组相比,各组的Ⅲ-C和PDGF m RNA表达有明显降低(P<0.01)。而各组中,灸药组疗效最明显,Ⅲ-C和PDGF的表达最低。蛋白免疫印迹法检测结果显示:与模型组相比各组的Ⅲ-C蛋白表达有差异。结论:1艾灸、化纤Ⅳ号方均可减轻博莱霉素诱导肺纤维化大鼠的肺纤维化程度。2艾灸配合化纤Ⅳ号方可减轻博莱霉素诱导肺纤维化大鼠的肺纤维化程度,且其效果优于单用艾灸或单用化纤Ⅳ号方。3艾灸、化纤Ⅳ号方及其二者配合使用不同程度阻抑博莱霉素诱导肺纤维化大鼠肺纤维化进程的效应机制,可能与通过调控其EMT过程中的Ⅲ-C和PDGF表达环节紧密相关。

1型糖尿病大鼠肾脏病变与ADPN、TGF-β1、collagen Ⅳ、ICAM-1的关系

目的探讨脂联素、TGF-β1、collagenⅣ、ICAM-1在1型糖尿病肾脏病变发生及发展中的变化及作用。方法大鼠随机分为正常对照组和糖尿病组,每组分别于4周和8周,测24h尿白蛋白排泄量(UPro/24h)、空腹血浆葡萄糖、血肌酐、尿素氮、血清脂联素(ADPN)、肾重/体重,观察肾脏TGF-β1、Ⅳ型胶原蛋白(collagenⅣ)、ICAM-1的表达。结果糖尿病大鼠UPro/24h增高、TGF-β1、collagenⅣ、ICAM-1的表达增加、脂联素水平降低。结论 TGF-β1、collagenⅣ、ICAM-1在肾脏组织中表达水平与血清ADPN呈负相关。随着ADPN水平降低,TGF-β1、collagenⅣ、ICAM-1在肾脏组织的表达增加。

Collagen type Ⅳ对周围神经中再生轴突及非神经元细胞的作用和影响

本文用抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ对抗体阻断ｃｏｌｌａｇｅｎｔｙｎｅⅣ的方法，研究了ｃｏｌｌａｇｅｎｔｙｐｅⅣ失活的移植神经段（长１０ｍｍ）植入大鼠坐骨神经后对再生轴突和非神经元细胞的作用和影响．实验结果显示：在移植神经段近端距近侧吻合口１ｍｍ处，术后１０ｄ抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ组再生轴突数为对照组的５４％，术后１５ｄ增加到６６％，术后３０ｄ高达９４％．在移植神经段远侧距近侧吻合口９ｍｍ处，术后３０ｄ抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ组再生轴突数为对照组的５８％。表明抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ组再生轴突的生长启动和生长速度明显慢于对照组．巨噬细胞在移植神经段内的滞留数量抗ｃｏｌｌａｇｅｎｔｙｐｅⅣ组明显多于对照组．这些结果揭示ｃｏｌｌａｇｅｎｔｙｐｅⅣ在神经损伤和再生中对促进轴突的生长和维持神经微环境的平衡起着积极的作用．本文对ｃｏｌｌａｇｅｎｔｙｐｅⅣ在神经再生中的作用机制作了初步的分析和探讨。

血管球瘤中FLi-1、h-caldesmon及collagen Ⅳ的表达及临床病理分析

目的探讨FLi-1、h-caldesmon及collagenⅣ在血管球瘤中的表达及不同组织学类型与临床病理指标的关系。方法采用免疫组化En Vision法检测35例血管球瘤中FLi-1、h-caldesmon及collagenⅣ的表达。分析35例血管球瘤不同组织学分型与临床病理参数间的关系。结果镜下见肿瘤细胞圆形或多边形,呈片状分布在血管之间或呈环状围绕在血管周围,瘤细胞边界清晰,外形规则,有时可见瘤细胞与梭形平滑肌细胞移行过渡。免疫表型:血管球瘤可表达FLi-1、h-caldesmon及collagenⅣ,阳性率分别为58.6%、97.1%和100%。血管球瘤的不同类型与患者性别和肿瘤体积无关,与患者年龄相关(P=0.01)。35例血管球瘤中vimentin和SMA均呈(+),2例CD34呈局灶(+),1例desmin呈(+),EMA、S-100、Cg A、CD68及CD99均呈(-)。结论 h-caldesmon和collagenⅣ可作为血管球瘤诊断的标志物,FLi-1可作为一种辅助参考标志物,有助于血管球瘤的诊断。

Sublytic C5b-9诱导肾小球系膜细胞产生的TSP-1和TGF-β1对其合成FN和collagenⅣ的影响

目的:探讨亚溶解型(sublytic)C5b-9复合物刺激大鼠肾小球系膜细胞(glomerular mesangial cell,GMC)后,诱生的血小板反应蛋白-1(thrombospondin-1,TSP-1)与转化生长因子-β1(transforming growth factor-β1,TGF-β1)对其促进细胞外基质(ex-tracellular matrix,ECM),如纤维连接蛋白(fibronectin,FN)和Ⅳ型胶原蛋白(collagenⅣ)合成的影响。方法:体外培养大鼠GMC,进行不同分组处理并给予sublytic C5b-9刺激,然后分别检查受刺激的GMC合成TSP-1和TGF-β1因子水平,同时检测GMC分泌FN和collagenⅣ的水平。此外,应用人工合成的TSP-1封闭肽段(GGWSHW)和TGF-β1中和抗体处理培养的大鼠GMC,研究TSP-1和TGF-β1在sublytic C5b-9诱导的GMC分泌上述ECM中的作用及其相互关系。结果:培养的大鼠GMC在sublytic C5b-9刺激后18 h,其FN和collagenⅣ表达水平均明显升高。同时,TSP-1蛋白表达和TGF-β1分泌(包括活化的TGF-β1含量)也显著增多。用TSP-1封闭肽段(GGWSHW)处理大鼠GMC后亦能显著抑制由sublytic C5b-9诱导的TGF-β1活化,并减少FN、collagenⅣ的产生。同样用TGF-β1中和抗体处理GMC后也能明显抑制由sublytic C5b-9导致的FN、collagenⅣ的分泌。结论:体外用sublytic C5b-9刺激大鼠GMC,能诱导其ECM分泌与TSP-1和TGF-β1的合成及活化,而sublytic C5b-9促进GMC分泌ECM的机制可能与其诱导TSP-1合成及活化的TGF-β1存在一定的关系。

芡实对糖尿病肾病大鼠肾组织MMP-9、TIMP-1及Collagen Ⅳ表达的影响

目的:探讨芡实对糖尿病肾病大鼠肾组织MMP-9、TIMP-1及CollagenⅣ表达的影响。方法:健康雄性Wistar大鼠65只,随机选取10只为正常对照组(N组);其余一次性腹腔注射链脲佐菌素(STZ)制作DN模型,造模成功后,随机分为DN组、芡实低、中、高剂量组及氯沙坦钾组,均采用灌胃给药,N组和DN组给予等体积蒸馏水。12周后检测大鼠生化指标;HE、Masson、PAS染色观察肾脏病理变化;免疫组化法测定MMP-9、TIMP-1及CollagenⅣ在肾组织表达情况;采用实时荧光定量PCR技术检测肾组织中MMP-9、TIMP-1及CollagenⅣ基因的表达。结果:实验12周末,与N组比较,DN组大鼠肾小球肥大,细胞外基质积聚,血糖、24 h尿蛋白定量、血尿素氮(BUN)、肌酐(Scr)、尿白蛋白排泄率(UAER)、尿蛋白/尿肌酐(Up/Ucr)比值显著升高(P<0.05),肾组织MMP-9蛋白和mRNA表达降低(P<0.05),TIMP-1、CollagenⅣ蛋白和mRNA表达升高(P<0.05);与DN组比较,EM组、EH组及LP组大鼠病理改变减轻,24 h尿蛋白定量、Scr、BUN、UAER、Up/Ucr显著降低(P<0.05),肾组织MMP-9蛋白和mRNA表达显著升高(P<0.05),TIMP-1、CollagenⅣ蛋白和mRNA表达显著减少(P<0.05)。结论:芡实可能通过调节DN大鼠肾脏MMP-9、TIMP-1的表达,减少细胞外基质积聚,从而减轻肾小球硬化,减少蛋白尿,保护肾功能,延缓DN病变的进展。

未知病变类型脑血管中β-amyloid、α-actin、collagen Ⅳ的含量

目的研究未知病变类型脑血管病变的结构特征。方法通过刚果红染色、免疫组织化学染色、计算机图像分析技术对未知病变类型脑血管病变的β-amyloid、α-actin、collagenⅣ的含量进行研究。结果未知病变类型脑血管壁α-actin、collagenⅣ呈少量阳性染色,与正常脑血管存在显著差异(P<0.05);β-amyloid染色呈阴性,与正常脑血管无差异(P>0.05)。病变血管壁中上述三种蛋白的表达特点与脑血管淀粉样变(cerebralamyloidangiopathy,CAA)及小动脉硬化玻璃样变不同。结论未知病变类型脑血管病变具有不同于CAA的病变特征。

Serum hyaluronic acid, procollagen type Ⅲ and Ⅳ in histological diagnosis of liver fibrosis

OBJECTIVE: To assess the significance of serum hyaluronic acid (HA), proeollagen type Ⅲ (PCⅢ), collagen type Ⅳ (CⅣ) in the histological diagnosis of liver fibrosis. METHODS: The concentrations of serum HA, PCⅢ, CⅣ in 253 patients with chronic liver diseases were measured by radioimmunoassay. Liver biopsies were performed in all patients at the same time. The liver was pathologically evaluated by a pathologist according to a scoring system. Combined with the results of liver pathological diagnosis, the accuracy of serum HA, PCⅢ, CⅣ in diagnosing patients with hepatic fibrosis (staging≥S_2) or cirrhosis (S_4) was assessed using the receiver operating curve (ROC). RESULTS: The cutoff values of serum HA, PCⅢ and CⅣ for identifying patients with hepatic fibrosis (≥S_2) or cirrhosis (S_4) were determined. The cutoff values of serum HA, PCⅢ and CⅣ for detecting patients with fibrosis (stage≥S_2) were 90μg/L, 90μg/L, 75μg/L, respectively; their sensitivity (Se) was 80.4%, 82%, 63.1%; their specificity (Spe) was 70.2%, 60.8%, 83.8%; their positive predictive values (PPV) were 86.7%, 83.5%, 90.4%; their negative predictive values (NPV) were 59.8%, 58.4%, 48.4%, respectively. The cutoff values for detecting patients with liver cirrhosis were 210μg/L for HA, 96.2% for Se, 85.3% for Spe, 65.4% for PPV, 98.8% for NPV; 150μg/L for PCⅢ, 76.4% for Se, 68.7% for Spe, 40.4% for PPV, 91.3% for NPV; 90μg/L for CⅣ, 80% for Se, 75.8% for Spe, 47.8% for PPV, 93.2% for NPV, respectively. CONCLUSIONS: Serum HA, PCⅢ and CⅣ can be determined for an accurate diagnosis of hepatic fibrosis in various stages. HA is the best for screening liver cirrhosis.

Effect of Marine Collagen Peptides on Markers of Metabolic Nuclear Receptors in Type 2 Diabetic Patients with/without Hypertension

Objective To explore Effects of marine collagen peptides （MCPs） on markers of metablic nuclear receptors, i.e peroxisome proliferator-activated receptor （PPARs）, liver X receptor （LXRs） and farnesoid X receptor （FXRs） in type 2 diabetic patients with/without hypertension. Method Study population consisted of 200 type 2 diabetic patients with/without hypertension and 50 healthy subjects, all of whom were randomly assigned to MCPs-treated diabetics （n=50）, placebo-treated diabetics （n=50）, MCPs-treated diabetics with hypertension （n=50）, placebo-treated diabetics with hypertension （n=50）, and healthy controls （n=50）. MCPs or placebo （water-soluble starch） were given daily before breakfast and bedtime over three months. Levels of free fatty acid, cytochrome P450, leptin, resistin, adiponectin, bradykinin, NO, and Prostacyclin were determined before intervention, and 1.5 months, and 3 months after intervention. Hypoglycemia and the endpoint events during the study were recorded and compared among the study groups. Result At the end of the study period, MCPs-treated patients showed marked improvement compared with patients receiving placebo. The protection exerted by MCPs seemed more profound in diabetics than in diabetics with hypertension. In particular, after MCPs intervention, levels of free fatty acid, hs-CRP, resistin, Prostacyclin decreased significantly in diabetics and tended to decrease in diabetic and hypertensive patients whereas levels of cytochrome P450, leptin, NO tended to decrease in diabetics with/without hypertension. Meanwhile, levels of adiponectin and bradykinin rose markedly in diabetics following MCPs administration. Conclusion MCPs could offer protection against diabetes and hypertension by affecting levels of molecules involved in diabetic and hypertensive pathogenesis. Regulation on metabolic nuclear receptors by MCPs may be the possible underlying mechanism for its observed effects in the study. Further study into its action may shed light on development of new drugs based on bioactive peptides from marine sources.

Adhesive properties of hepatoma cells to collagen Ⅳ coated surfaces

Objectives: To quantitatively study the adhesive pro- perties of hepatoma cells to collagen Ⅳ coated artifi- cial basement membrane and to investigate the rele- vance of cell adhesive forces to the concentration of collagen Ⅳ. Methods: Synchronous G1 and S phase cells were a- chieved using thymine-2-desoxyriboside and cochicine sequential blockage method and double thymine-2- desoxyriboside blockage method respectively. The adhesive forces of hepatoma cells were investigated by micropipette aspiration technique. Results: The adhesive forces of hepatoma cells to ar- tificial basement membrane were (107.78±65.44) ×10^(-10)N, (182.60±107.88)×10^(-10)N, (298.91± 144.13)×10^(-10)N when the concentration of the membrane coated by 1, 2, 5μg/ml collagen Ⅳ re- spectively (P<0.001). The adhesive forces of G1 and S phases hepatoma cells to artificial basement membrane were (275.86±232.80)×10^(-10)N and (161.16±120.40)×10^(-10)N respectively when the concentration of the membrane coated by 5μg/ml collagen Ⅳ (P<0.001). Conclusions: The adhesive forces of hepatoma cells to artifical basement membrane in direct proportion to the concentration of collagen Ⅳ suggests that the in- crease of basement membrane might be conducive to the chemotactic motion and adhesiveness of tumor cells. G1 phase cells are more capable of adhering to basement membrane than S phase cells. Hepatoma cells, especially G1 phase cells, may survive in blood circulation, and sequest and adhere in microcircula- tion, and get through basement membrane for re- mote metastasis.

Analysis of Metabolic Products by Response Surface Methodology for Production of Human-like Collagen II

Recombinant Escherichia coli BL21 is used to produce human-like collagen. The key constituents of media are optimized using response surface methodology (RSM). Before thermal induction, the highest biomass production and the lowest production of some hazardous by-products, especially acetic acid, were obtained in the media containing 0.085 mol·L-1 glucose and 0.019 mol·L-1 nitrogen (carbon-nitrogen ratio, 4.47:1). After thermal induction, when the concentrations of glucose and nitrogen in the media were 0.065 mol·L-1 and 0.017 mol·L-1 , respectively (carbon-nitrogen ratio, 3.82:1), the productivity of human-like collagen per cell was the highest while that of acetic acid was the lowest. The extended analysis showed that the production of lactic acid and propionic acid increased while that of some intermediate acids of the tricarboxylic acid cycle decreased if the dose of glucose increased.

Expression of Collagen Ⅳ, Fibronectin, Laminin in Non-small Cell Lung Cancer and Its Correlation with Chemosensitivities and Apoptosis~*

Objective： To study the expression of extracellular matrix （ECM） proteins including Collagen Ⅳ （Co Ⅳ）, Fibronectin, Laminin in human non-small cell lung cancer （NSCLC） specimens and the relationship between ECM and cell adhesion, proliferation, apoptosis and drug sensitivity in NSCLC cell line. And to investigate the role of phosphatidylinositol 3-kinase （PI3-K） in signal transduction of Co Ⅳ in NSCLC. Methods： The expression of ECM proteins was detected by using immunohistochemical staining （Envision＇s）. Adherent cells were stained with 1% methylene blue. Cell proliferation and cytotoxic effects were monitored by MTT assay. Cell apoptosis was analyzed by FITC-Annexin V/PI double staining variables flow cytometry （FCM）. Results： The expression rate of Co Ⅳ （93%） was the highest compared to others in NSCLC stroma. After treated with Co Ⅳ, the adhesion of H1299 cells was increased and the cytotoxicity of cis-platinum （DDP） against H1299 cells was decreased compared to the control （P〈0.05）. After treated with Co Ⅳ both survival and proliferation rates were higher and apoptosis rate was lower than without Co Ⅳ （P〈0.05）. PI3-K inhibitor LY294002 decreased both survival and proliferation rates （82.7%±2.0% and 75.2%±6.8%, respectively）, even on Co Ⅳ-coated surface （92.2%±2.8% and 84.6%±9.2%, respectively）. And it also helped DDP increase apoptosis. Conclusion： ECM remodeling existed in NSCLC. Co Ⅳ protected NSCLC cells from DDP-induced apoptosis and weakened the cytotoxicity of DDP. PI3-K pathway might be the crucial mechanism of apoptosis impairment and drug resistance.

Serum type Ⅳ collagen level is predictive for esophageal varices in patients with severe alcoholic disease

AIM： To determine factors predictive for esophagea varices in severe alcoholic disease （SAD）. METHODS： Abdominal ultrasonography （US） was performed on 444 patients suffering from alcoholism. Forty-four patients found to have splenomegaly and/ or withering of the right liver lobe were defined as those with SAD. SAD patients were examined by upper gastrointestinal （UGI） endoscopy for the presence of esophageal varices. The existence of esophageal varices was then related to clinical variables. RESULTS： Twenty-five patients （56.8%） had esophageal varices. A univariate analysis revealed a significant difference in age and type Ⅳ collagen levels between patients with and without esophageal varices. A logistic regression analysis identified type Ⅳ collagen as the only independent variable predictive for esophageal varices （P = 0.017）. The area under the curve （AUC） for type Ⅳ collagen as determined by the receiver operating characteristic （ROC） for predicting esophageal varices was 0.78. CONCLUSION： This study suggests that the level of type Ⅳ collagen has a high diagnostic accuracy for the detection of esophageal varices in SAD.

Pre-hepatectomy type Ⅳ collagen 7S predicts post-hepatectomy liver failure and recovery

BACKGROUND Liver resection is an effective treatment for benign and malignant liver tumors.However,a method for preoperative evaluation of hepatic reserve has not yet been established.Previously reported assessments of preoperative hepatic reserve focused only on liver failure in the early postoperative period and did not consider the long-term recovery of hepatic reserve.When determining eligibility for hepatectomy,the underlying pathophysiology needs to be considered to determine if the functional hepatic reserve can withstand both surgery and any postoperative therapy.AIM To identify pre-hepatectomy factors associated with both early postoperative liver failure and long-term postoperative liver function recovery.METHODS This study was a retrospective cohort study.We retrospectively investigated 215 patients who underwent hepatectomy at our hospital between May 2013 and December 2016.Early post-hepatectomy liver failure(PHLF)was defined using the International Study Group of Liver Surgery’s definition of PHLF.Long-term postoperative recovery of liver function was defined as the time taken for serum total bilirubin and albumin levels to return to levels of<2 mg/dL and>2.8 g/dL,respectively,and the time taken for Child-Pugh score to return to Child-Pugh class A.RESULTS Preoperative type IV collagen 7S was identified as a significant independent factor associated with both PHLF and postoperative long-term recovery of liver function.Further analysis revealed that the time taken for the recovery of Child-Pugh scores and serum total bilirubin and albumin levels was significantly shorter in patients with type IV collagen 7S≤6 ng/mL than in those with type IV collagen 7S>6 ng/mL.In additional analyses,similar results were observed in patients without chronic viral hepatitis associated with fibrosis.CONCLUSION Preoperative type IV collagen 7S is a preoperative predictor of PHLF and longterm postoperative liver function recovery.It can also be used in patients without chronic hepatitis virus.

Effects of Calcium Dobesilate on Glomerulus TIMP1 and Collagen Ⅳ of Diabetic Rats

Summary ： To observe the effects of calcium dobesilate on the expression of glomerular tissue inhibitor of metalloproteinase 1 （TIMP1）, collagen Ⅳ , and ultrastrueture of glomerular basement mem- brane in diabetic rats, rats model of diabetes was established by unilateral nephreetomy and intraperitoneal injection of 1% STZ （55 mg/kg）, and rats were administered calcium dobesilate 100 mg/ kg （DD group） or distilled water （DM group） respectively. 12 weeks later, the changes in the renal uhrastrueture and ereatinine clearance rate （Cer） were examined in each group. The expression of glomerular TIMP1 and collagen Ⅳ were studied by immunohistoehemieal staining. Our results showed that after 12 weeks, the Cer in DD group increased and was significantly higher than that in DM group. Electron microscopy showed that thickness of glomerular capillary basement membrane （GBM） in Group DD was less than that of DM group. No hyperplasia of collagen fibers was found, and the distance betweeh the holes of endothelial cells in DD group was not as even as that in the normal group, but more even than that of DM group, and podocyte processes was still in order. Immunohistochemical staining of glomeruli showed that expression of TIMP1 and collagen Ⅳ in DD group were significantly less than those of DM group DM. It is concluded that calcium dobesilate can improve diabetic nephropathy by inhibiting the overaccumulation of collagen Ⅳ and calcium dobesilate may also contribute to diabetes by inhibiting the expression of TIMP1.

The Effectiveness of Specific Collagen Peptides on Osteoarthritis in Dogs-Impact on Metabolic Processes in Canine Chondrocytes

In clinical trials over the past decade, the beneficial effect of orally administered collagen peptides in osteoarthritic dogs has been clearly demonstrated [1] [2] [3]. Although a statistically significant improvement in the lameness and vitality of dogs in general has been documented, the mode of action of the collagen peptide treatment is still under discussion. A previous study [3] indicated that the reduction in lameness and increased mobility in dogs after collagen peptide treatment were associated with a statistically significantly lowered plasma content of MMP-3, which is involved in collagen degradation. In addition, the content of the MMP-antagonist TIMP-1 increased slightly after collagen peptide supplementation, suggesting a direct impact on the cartilage metabolism, particularly on the decrease of extracellular matrix degradation. Based on these findings, the impact of specific collagen peptides (PETA-GILE?) on cartilage metabolism was tested in canine chondrocytes in the current investigation. In addition to the biosynthesis of various matrix molecules (type II collagen, aggrecan and elastin), the RNA profile of inflammatory cytokines and degenerative matrix molecules was investigated. The results showed clearly that the supplementation of specific collagen peptides reduced catabolic processes, as indicated by a statistically significant decrease in inflammatory cytokines and proteases in canine chondrocytes compared with untreated control experiments. In addition, a statistically significantly enhanced biosynthesis of type II collagen, elastin, and aggrecan was observed. Hence, the current data supports the suggested anti-inflammatory effect of specific collagen peptides, but also clearly demonstrates a pronounced stimulatory impact on matrix molecule synthesis. A combination of both observed effects might help to explain the previously reported clinical improvements after collagen peptide supplementation. Furthermore, the beneficial effect of the specific collagen peptides was also confirmed in case reports on osteoarthritic dogs that demonstrated decreased lameness and increased vitality in the affected animals after PETAGILE treatment.