Expression of insulin-like growth factor Ⅱ and its receptor in liver cells of chronic liver diseases

AIM To clarify the relationship between the Insulin like growth factor Ⅱ (IGF Ⅱ), IGF Ⅱ receptor and chronic liver diseases and to provide evidences for basic and clinical researches for exploring the potential mechanisms of human hepatocellular carcinoma (HCC). METHODS The poly (A)+ mRNA translation of IGF Ⅱ and IGF Ⅱ receptor in dysplasia liver cell (DLC n =10), liver cirrhosis (LC n =9) and chronic active hepatitis (CAH n =9) were analyzed with RNA gel electrophoresis, Northern blot and hybridization using human IGF Ⅱ and IGF Ⅱ receptor DNA probes labelled with 32 P through Nick translation and autoradiography. RESULTS The overexpression of IGF Ⅱ in DLC (10/10, 100%) was apparently higher than that in CAH (3/9, 33%) and LC (3/9, 33%), ( P <0 01). The overexpression of IGF Ⅱ receptor in DLC (7/10, 70%) was significantly higher than that in CAH (2/9, 22%) and LC (3/9, 33%), respectively. The data of HBV infection from different chronic liver diseases were analyzed. CONCLUSION The overexpression of IGF Ⅱ and IGF Ⅱ receptor in DLC was related to the preceeding of malignant phenotype of hepatocyte, which provided a diagnostic value for early detection of hepatocellular carcinoma (HCC). Persistent HBV infection is strongly associated with abnormal activation of IGF Ⅱ and IGF Ⅱ receptor, which might indicate a stimulating mechanism of autocrine or paracrine growth involved in live cell carcinogenesis.

Interplay between micro RNA-17-5p, insulin-like growth factor-Ⅱ through binding protein-3 in hepatocellular carcinoma

AIM: To investigate the effect of microR NA on insulinlike growth factor binding protein-3(IGFBP-3) and hence on insulin-like growth factor-Ⅱ(IGF-Ⅱ) bioavailability in hepatocellular carcinoma(HCC).METHODS: Bioinformatic analysis was performed using microrna.org, DIANA lab and Segal lab softwares. Total RNA was extracted from 23 HCC and 10 healthy liver tissues using mir Vana mi RNA Isolation Kit. microR NA-17-5p(miR-17-5p) expression was mimicked and antagonized in Hu H-7 cell lines using Hi Per Fect Transfection Reagent, then total RNA was extracted using Biozol reagent then reverse transcribed into cD NA followed by quantification of mi R-17-5p and IGFBP-3 expression using Taq Man real-time quantitative PCR. Luciferase reporter assay was performed to validate the binding of miR-17-5p to the 3'UTR of IGFBP-3. Free IGF-Ⅱ protein was measured in transfected Hu H-7 cells using IGF-Ⅱ ELISA kit. RESULTS: Bioinformatic analysis revealed IGFBP-3 as a potential target for miR-17-5p. Screening of miR-17-5p and IGFBP-3 revealed a moderate negative correlation in HCC patients, where mi R-17-5p was extensively underexpressed in HCC tissues(P = 0.0012), while IGFBP-3 showed significant upregulation in the same set of patients(P = 0.0041) compared to healthy donors. Forcing mi R-17-5p expression in Hu H-7 cell lines showed a significant downregulation of IGFBP-3 mR NA expression(P = 0.0267) and a significant increase in free IGF-Ⅱ protein(P = 0.0339) compared to mock untransfected cells using unpaired t-test. Luciferase assay validated IGFBP-3 as a direct target of mi R-17-5p; luciferase activity was inhibited by 27.5% in cells co-transfected with miR-17-5p mimics and the construct harboring the wild-type binding region 2 of IGFBP-3 compared to cells transfected with this construct alone(P = 0.0474).CONCLUSION: These data suggest that regulating IGF-Ⅱ bioavailability and hence HCC progression can be achieved through targeting IGFBP-3 via manipulating the expression of miR NAs.

Expression of insulin-like growth factor Ⅱ and its receptor in hepatocellular carcinogenesis

INTRODUCTIONInsulin-like growth factor Ⅱ(IGF-Ⅱ) is a mitogenic peptide of 74 kD and is mostly synthesized in fetal liver tissue .IGF-Ⅱ is believed to play an important role in fetal growth and development and is involved in cellular proliferation and differentiation[1-5]. Recently ,several researchers have reported increased expression of the IGF-Ⅱgene in human hepatocellular carcinoma (HCC) and adjacent non-cancerous liver tissues [6-10].

The immunocytochemical localization of insulin-like growth factor Ⅱ in human cirrhosis

Sections of 30 cases of human cirrhosis were stained with rabbit anti-insulin-likegrowth factor Ⅱ(IGF Ⅱ)by double PAP method.By the serological examination 15 patientsshowed HBV infection and sections of 14 eases were HBsAg postively with a total rate of 67％(20 cases).The IGF Ⅱ was positive in the cytoplasm of all the liver and ductular cells.Binucle-ated,polypoid liver cells and the peripheral cells of the lobules or nodules were distinctly posi-tive,The liver cells which were strongly positive were a kind of thin polygonal cells with asmall oval or a round deeply stained nucleus in each.They might exist sporadically in the lob-ules or in the marginal portion of a nodule.These liver cells are quite different from the so-called oval cells which are derived from the proliferating ductules and are generally believed tobe responsible for the pathogensis of hepatoma.

Changes of insulin-like growth factor-Ⅱ and insulin-like growth factor binding protein-3 in cerebrospinal fluid of children with tuberculous meningitis

BACKGROUND： Recent studies have found that insulin-like growth factors （IGFs） and insulin-like growth factor binding protein-3 （IGFBP-3） have stronger neurotrophic and neuroprotective effects. But whether their levels in cerebrospinal fluid could be used as an auxiliary indicator in differentially diagnosing tuberculous meningitis and viral encephalitis is not yet clear. OBJECTIVE： To explore the changes of insulin-like growth factor-Ⅱ （IGF-Ⅱ ） and IGFBP-3 in cerebrospinal fluid （CSF） of children with tuberculous meningitis and the significance of the changes. DESIGN： A non-randomized concurrent controlled study. SETTING： Department of Pediatric Internal Medicine, the First Affiliated Hospital of Xinxiang Medical College. PARTICIPANTS： Thirty children with tuberculous meningitis （14 males and 16 females） were selected from the Department of Pediatric Internal Medicine, the First Affiliated Hospital of Xinxiang Medical College from January 2005 to December 2006. Tuberculous meningitis was diagnosed according to their clinical manifestations, the history of close contact with tuberculosis, typical cerebrospinal fluid changes of tuberculous meningitis, positive tuberculosis antibody and effective antituberculosis treatment. There were 30 children （13 males and 17 females） with viral encephalitis, and viral encephalitis was diagnosed according to epidemiological history, clinical manifestations, conventional and biochemical changes of cerebrospinal fluid, and negative bacteriology judgment. Meanwhile, 30 children （13 males and 17 females） without infectious and central nervous system disease were selected as the control group. Informed consent was obtained from the parents of all the enrolled children. METHODS： ①The lumbar puncture operation was implemented immediately to obtain cerebrospinal fluid （3 mL）. The contents of IGF-Ⅱ and IGFBP-3 were detected with immunoradiometric assay. The concentrations of glucose and protein in cerebrospinal fluid were determined with a dry-chemical method. The number of white blood cells was counted by Fushi Method. ②The Pearson correlation analysis was used to analyze the correlation of the contents of IGF-Ⅱ and IGFBP-3 in cerebrospinal fluid with the leucocyte counting and the concentrations of glucose and protein in cerebrospinal fluid. MAIN OUTCOME MEASURES： The contents of IGF- Ⅱ and IGFBP-3 in cerebrospinal fluid, and their correlation with the leucocyte counting and the concentrations of glucose and protein in cerebrospinal fluid. RESULTS： ①Contents of IGF-Ⅱ and IGFBP-3 in cerebrospinal fluid： The contents of IGF-Ⅱ and IGFBP-3 in cerebrospinal fluid in the tuberculous meningitis group were significantly higher than those in the encephalitis virus group and control group （P 〈 0.05）. There was no significant difference in the contents of IGF- Ⅱ and IGFBP-3 in cerebrospinal fluid between the viral encephalitis group and control group （P 〉 0.05）. ②Correlation： The IGF- Ⅱ and IGFBP-3 contents in cerebrospinal fluid were positively correlated with the protein concentration in cerebrospinal fluid （r =0.821, 0.855, P 〈 0.01）, but negatively with the glucose （r =0.742, - 0.605, P 〈 0.01）. CONCLUSION- ①IGFs and IGVBPs are involved in the pathophysiological process of tuberculous meningitis, as well as the glucose and protein metabolism in cerebrospinal fluid. ②The IGF-Ⅱ and IGFBP-3 contents in cerebrospinal fluid can be used as the auxiliary indicators to differentially diagnose tuberculous meningitis and viral enceohalitis.

Antisense oligonucleotide to insulin-like growth factorⅡ induces apoptosis in human ovarian cancer AO cellline

The effects of antisense oligonucleotide to insulin-like growth factor 11 (IGFII) to induce apoptosis in human ovarian cancer cells were evaluated. Antiproliferation effects of antisense to IGFII in ovarian cancer AO cells were determined by 3H-thymidine incorporation. Apoptosis of the IGFll antisense-treated cells was quantitated by both nuclear condensation and flow cytometry after cells were stained with propidium iodide. IGFII antisense (4.5μM)treatment of 48 h maximally inhibited proliferation of AO cells. More than 25% of IGFII antisense-treated cells (4.5PM for 24 h) had undergone apoptosis, whereas less than 3% of the cells were apoptotic in either IGFII sense-treatedcells or untreated cells. Antisense oligonucleotide to IGFII significantly inhibited cell proliferation and induced apoptosis in human ovarian cancer AO cell. These data suggest that IGFII may be a potential target in treatment of ovarian cancer and antisense oligonucleotide to IGFⅡmay serve as a therapeutic approach.

母乳IGF-Ⅱ与婴儿体格生长指标关联性分析及影响因素研究

目的描述哺乳期各阶段母乳IGF-Ⅱ浓度的变化,探讨哺乳期不同阶段母乳IGF-Ⅱ与婴儿体格生长指标的关联,以及可能影响母乳IGF-Ⅱ浓度的因素。方法2020年10月至2021年9月于北京大学人民医院招募32名满足纳入排除标准的分娩妇女,分别于分娩后48 h、产后15 h、产后42 h、6个月、9个月及12个月采集妇女母乳标本,同时收集婴儿身长、体重等生长发育指标。采用酶联免疫吸附方法检测母乳IGF-Ⅱ水平。使用广义估计方程(GEE)分析母乳IGF-Ⅱ水平与婴儿体格生长指标之间的关联以及可能影响母乳IGF-Ⅱ水平的因素。结果母乳IGF-Ⅱ中位浓度在12月成熟乳中最高(82.5 ng/mL),6月成熟乳中最低(55.3 ng/mL),呈先下降再升高趋势,但各阶段中位浓度间未见明显差异;在调整母亲年龄、孕前BMI、孕周、母亲民族、孕产史、孕期并发症情况(妊娠期糖尿病、妊娠期高血压、妊娠期甲状腺疾病)、婴儿喂养方式及婴儿性别后,研究结果显示初乳中IGF-Ⅱ含量与婴儿身长呈正相关关系(β=0.9,P<0.01),初乳、42d成熟乳、12月成熟乳中的IGF-Ⅱ含量与婴儿体重呈正相关关系(P=0.02,P<0.01和P=0.01)。将膳食炎症指数(DII)按三分位法分为三组(Q1:最抗炎倾向组,Q2:中间组,Q3:最促炎倾向组)纳入GEE模型,结果显示,第三分位组(即最促炎倾向组)与母乳IGF-Ⅱ含量正相关(β=28.6,P=0.01)。结论母乳IGF-Ⅱ对婴儿身长、体重存在一定的促进作用,孕期促炎饮食与母乳IGF-Ⅱ含量相关。

Effects of Tanshinone ⅡA on Transforming Growth Factor β1-Smads Signal Pathway in Renal Interstitial Fibroblasts of Rats

The effects of tanshinone ⅡA （TSN） on transforming growth factor β1 （TGFβ1） signal transduction in renal interstitial fibroblasts of rats were studied in order to investigate its mechanism in prevention of renal interstitial fibrosis. Rat renal fibroblasts of the line NRK/49F were cultured in vitro, stimulated with 5 ng/mL TGFβ1 and pretreated with 10-6, 10-5, 10-4 mol/L TSN respectively. The mRNA levels of fibronectin （FN） were examined by RT-PCR. The protein expression of FN and Smads was detected by Western blot. TGFβ1 induced the expression of FN mRNA and Smads in a time-dependent manner in a certain range. Compared with pre-stimulation, the FN mRNA and protein levels were increased by 1.1 times and 1.5 times respectively （P〈0.01, P〈0.01）, and the protein expression of phosphorylated Smad2/3 （p-Smad2/3） increased by 7 times at the end of TGFβ1 stimulation （P〈0.01）. TSN pretreatment may down-regulate the FN and p-Smad2/3 expression in a dose-dependent manner. 10-6 mol/L TSN pretreatment had no effect on the FN and p-Smad2/3 expression （both P〉0.05）. After pretreatment with 10-5 and 10-4 mol/L TSN, the FN mRNA levels were decreased by 28.1% and 43.8% respectively （P〈0.05, P〈0.01）, the FN protein levels were decreased by 40% and 44% respectively （P〈0.05, P〈0.05）, and the p-Smad2/3 protein expression were decreased by 40% and 65% respectively （P〈0.05, P〈0.01）. The inhibitory effect of TSN on renal interstitial fibrosis may be related to its blocking effect on TGFβ1-Smads signal pathway in renal intersti- tial fibroblasts.

Detection of Frameshift Mutations of the Transforming Growth Factor p ReceptorⅡin Gastric Cancers with Microsatellite Instability

OBJECTIVE To study the relationship among microsatellite instability （MSI）, frameshift mutations （FM） of the transforming growth factor β receptor Ⅱ （TGF β R Ⅱ）, methylation state of the hMLH1 promoter and hMLH1 protein expression level in gastric cancers, and to explore their relationship to gastric carcinogenesis. METHODS DNA was isolated from 101 gastric specimens and 5 microsatellite loci were detected. PCR, electrophoresis on denatured polyacrylamide gels and silver staining were performed to detect the MSI. The FMs of TGFβR Ⅱ were also screened with the same method. HMLH1 methylation was analyzed by methylation specific PCR （MSP） and sequencing. HMLH1 protein expression was detected using immunohistochemistry. RESULTS The incidence of MSIs was 53.7% and 0% in the cancers and normal tissues, respectively, with the frequency of MSIs being significantly higher in the gastric cancers compared to the normal gastric tissues （P〈0.05）. The frequency of hMLH1 methylation was 41.5%（17/41） in the gastric cancers and 0%（0/60） in the normal group. Decreased hMLH1 expression was observed in 94.1%（16/17） of cases exhibiting methylation. FMs of TGFβR Ⅱ were identified in 5 （62.5%） of the 8 samples with MSIH. In contrast, FMs were not found in MSI-L or microsatellite stable （MSS） cases. CONCLUSION MSIs and FMs of TGFβR Ⅱ may play an important role in gastric carcinogenesis. HMLH1 methylation is an important modification of the DNA which results in inactivation of hMLH1 and mismatch repair defects which lead to MSls and FMs of TGFβR Ⅱ.

GDF15、miR-122、AFP和PIVKA-Ⅱ联合评估在HBV感染肝硬化患者肝细胞癌发生风险中的预测价值

目的评估血清生长分化因子15(GDF15)、microRNA-122(miR-122)、甲胎蛋白(AFP)和异常凝血酶原(PIVKA-Ⅱ)联合检测在监测乙肝病毒(HBV)感染肝硬化患者中肝细胞癌(HCC)的风险预测价值。方法将2017年12月至2022年1月在首都医科大学附属北京友谊医院就诊的45例HBV相关肝硬化患者、50例HCC患者纳入横断面研究,设为肝硬化组和HCC组;另选22例监测期间新确诊HCC的HBV肝硬化患者纳入纵向研究队列,分别对肝硬化组和HCC组患者以及监测期间新确诊HCC的HBV肝硬化患者的系列血清标本[新确诊HCC前12~18个月(T_(1))、新确诊HCC前6~12个月(T_(2))以及HCC诊断时(T_(3))]进行检测,测定GDF15、miR-122、AFP和PIVKA-Ⅱ水平。比较各组患者每项标志物的检测水平,采用受试者工作特征(ROC)曲线分析GDF15、miR-122、AFP和PIVKA-Ⅱ单独或联合检测对HCC患者的诊断价值,评估标志物的组合预测HBV感染肝硬化患者的HCC风险。结果横断面研究中,HCC组患者血清GDF15、AFP和PIVKA-Ⅱ水平分别为1760.64 pg/mL、40.24 ng/mL、106.37 mAU/mL,均显著高于肝硬化组患者(1357.63 pg/mL、9.07 ng/mL、22.59 mAU/mL),miR-122水平为38.72,则显著低于肝硬化组患者(75.70),差异均有统计学意义(P<0.05)。GDF15、miR-122、AFP和PIVKA-Ⅱ单独诊断HCC时,曲线下面积(AUC)分别为0.734、0.644、0.776、0.823;AFP和GDF15两项联合,AUC为0.835;GDF15、AFP和PIVKA-Ⅱ三项联合,AUC为0.860;GDF15、miR-122、AFP和PIVKA-Ⅱ四项联合,AUC最佳为0.876。区分肝硬化和HCC时,PIVKA-Ⅱ具有较高的敏感度(72.5%);GDF15具有较高的特异度(83.2%);AFP和PIVKA-Ⅱ联合,特异度最高(92.0%);四项联合,敏感度增加(82.2%),约登指数最高(0.622)。纵向研究结果未观察到每种单一生物标志物随时间的变化,而GDF15、AFP和PIVKA-Ⅱ三项联合以及GDF15、miR-122、AFP和PIVKA-Ⅱ四项联合,在三个时间点的检测值差异有统计学意义(P<0.01)。肝硬化患者横截面和纵向(T_(1))之间观察到四项组合的差异有统计学意义(P<0.05),提示标志物的联合应用可有效区分即将发生HCC和不会发生HCC的肝硬化患者。结论GDF15、miR-122、AFP和PIVKA-Ⅱ联合检测可以提高对HBV相关肝硬化和HCC患者的分辨力,且在HBV相关的肝硬化中,GDF15、miR-122、AFP和PIVKA-Ⅱ的组合能够识别HCC发展风险较高的患者,具有临床价值。

Transforming growth factor β receptor Ⅱ expression inexperimental cryptorchidism and apoptosisin spermatogenic cells in rats

Objective: To study the transforming growth factor β receptor Ⅱ (TGFβ-R Ⅱ) expression in experimental cryptorchidism and apoptosis in spermatogenic cells in rats. Methods: The apoptosis of spermatogenic cells was detected by means of the terminal deoxynucleotldyl transferase mediated dUTP nick end lableling method (TUNEL) and the TGFβ-R Ⅱ expression was observed with the immunohistochemistry SABC methods. Results: There was a significant increase in the TGFβ-R Ⅱ expression in unilateral undescended testes (UUTs) compared with that in contralateral descended testes (CDTs, P<0.01). However, there was a significant and time-dependent increase in the mean apoptotic index in UUTs than in CDTs. Conclusion: TGFβ-R Ⅱ may play an important role in spermatogenic cell apoptosis.

Ⅱ型子宫内膜癌患者癌组织中IMP3、PTEN表达及其临床意义

目的分析Ⅱ型子宫内膜癌(EC)患者癌组织中胰岛素样生长因子Ⅱ信使RNA结合蛋白3(IMP3)和磷酸酶与张力蛋白同源物(PTEN)的表达情况,并探讨其临床意义。方法收集60例Ⅱ型EC患者手术切除的癌组织及20例对应癌旁组织的存档石蜡标本,采用免疫组织化学法(IHC)检测标本中IMP3、PTEN蛋白表达情况;分析IMP3、PTEN蛋白表达与EC患者临床病理特征及预后的关系。结果EC患者癌组织中IMP3阳性表达率高于癌旁组织,PTEN阳性表达率低于癌旁组织,差异均有统计学意义(P<0.05)。EC患者癌组织IMP3阳性表达率与肌层浸润深度、FIGO分期及淋巴结转移有关(P<0.05),即肌层浸润深度越深、FIGO分期越晚、合并淋巴结转移者IMP3阳性表达率越高。PTEN阳性表达率与FIGO分期、淋巴结转移有关(P<0.05),即FIGO分期越晚、合并淋巴结转移者PTEN阳性表达率越低。Spearman相关性分析结果显示,EC患者癌组织中IMP3和PTEN蛋白表达无显著相关性(P>0.05)。IMP3阳性组整体生存情况差于IMP3阴性组,PTEN阳性组整体生存情况优于PTEN阴性组(P<0.05)。结论Ⅱ型EC患者癌组织中IMP3表达上调,而PTEN表达下调,二者表达异常可能参与了Ⅱ型EC的发生与发展,且IMP3阳性表达、PTEN阴性表达与Ⅱ型EC患者不良预后相关。

右美托咪定通过调控白细胞介素-17及转化生长因子-β1/Smad蛋白3信号通路对脂多糖诱导Ⅱ型肺泡上皮细胞损伤的影响

目的探讨右美托咪定介导白细胞介素(IL)-17对脂多糖(LPS)诱导的人Ⅱ型肺泡上皮A549细胞(以下简称“A549细胞”)炎症、增殖、迁移、上皮间质转化及转化生长因子-β1(TGF-β1)/Smad蛋白3(Smad3)信号通路的调控作用。方法体外培养A549细胞,将其分为对照组(不做干预)、LPS组(10.00μg/ml LPS)和实验组(10.00μg/ml LPS+1.25、2.50、5.00、10.00μg/ml右美托咪定),分别采用CCK-8试剂盒和反转录-聚合酶链反应(RT-qPCR)法测定细胞活力和IL-17 mRNA表达水平以筛选右美托咪定最适实验浓度;随后将细胞分为对照组、LPS组、IL-17组、右美托咪定组和IL-17+右美托咪定组,干预24 h。酶联免疫吸附试验(ELISA)检测炎症因子IL-17、IL-8和IL-6的表达水平;5-乙炔基-2'脱氧尿嘧啶核苷(EdU)试剂盒用来检测细胞增殖率;划痕实验用来检测细胞迁移率;蛋白免疫印迹(WB)法测定肺泡上皮间质转化(EMT)相关蛋白、TGF-β1/Smad3通路相关蛋白及IL-17蛋白表达水平。结果右美托咪定逆转了LPS对A549细胞活力的抑制作用和IL-17 mRNA表达水平的促进作用,且10.00μg/ml浓度的右美托咪定组的效果最好,因此选择10.00μg/ml右美托咪定用于后续实验。LPS组细胞中IL-17、IL-8、IL-6表达水平、细胞迁移率、N-钙黏蛋白、波形蛋白、纤维粘连蛋白(FN)、Smad3的磷酸化(p-Smad3)/Smad3、TGF-β1和IL-17蛋白表达水平高于对照组,差异有统计学意义(P<0.05);LPS组细胞增殖率、E-钙黏蛋白和Smad7蛋白表达水平低于对照组,差异有统计学意义(P<0.05)。IL-17+右美托咪定组IL-17、IL-8、IL-6表达水平、细胞迁移率、N-钙黏蛋白、波形蛋白、FN、p-Smad3/Smad3、TGF-β1和IL-17蛋白表达水平低于LPS组,差异有统计学意义(P<0.05);IL-17+右美托咪定组细胞增殖率、E-钙黏蛋白和Smad7蛋白表达水平高于LPS组,差异有统计学意义(P<0.05)。结论右美托咪定通过抑制IL-17的分泌恢复LPS诱导的人Ⅱ型肺泡上皮细胞损伤,促进LPS诱导的人Ⅱ型肺泡上皮细胞增殖并抑制其迁移和EMT进程,其作用机制与抑制TGF-β1/Smad3通路的信号转导有关。

羟基红花黄色素A抑制血管紧张素Ⅱ介导的心脏成纤维细胞肌化

目的研究羟基红花黄色素A对血管紧张素Ⅱ介导的心脏成纤维细胞向肌成纤维细胞转化的影响。方法实验细胞随机分为正常对照组、Ang-Ⅱ组、Ang-Ⅱ+HSYA(5μM)组、Ang-Ⅱ+HSYA(25μM)组、Ang-Ⅱ+HSYA(50μM)组和Ang-Ⅱ+HSYA(100μM)组。使用划痕实验和Transwell小室侵袭实验检测细胞迁移侵袭情况,DCFH-DA荧光探针检测细胞内活性氧(reactive oxygenspecies,ROS)水平,Western blot检测α-SMA、CollagenⅠ、CollagenⅢ及转化生长因子β1(transforming growthfactor-β1,TGF-β1)的蛋白表达水平。结果Ang-Ⅱ促进心脏成纤维细胞迁移、增加ROS产生;而HSYA抑制了Ang-Ⅱ介导的心脏成纤维细胞迁移以及ROS产生;Western blot发现HSYA抑制了Ang-Ⅱ介导的心脏成纤维细胞α-SMA、CollagenⅠ、CollagenⅢ及TGF-β1的蛋白表达,差异具有统计学意义(P<0.05)。结论HSYA通过减少ROS的产生和下调TGF-β1信号通路抑制Ang-Ⅱ诱导心肌成纤维细胞向肌成纤维细胞分化。

Intestinal hormones and growth factors:Effects on the small intestine

There are various hormones and growth factors which may modify the intestinal absorption of nutrients, and which might thereby be useful in a therapeutic setting, such as in persons with short bowel syndrome. In part I, we focus first on insulin-like growth factors, epidermal and transferring growth factors, thyroid hormones and glucocorticosteroids. Part Ⅱ will detail the effects of glucagon-like peptide （GLP）-2 on intestinal absorption and adaptation, and the potential for an additive effect of GLP2 plus steroids.

GubenyiliuⅡ inhibits breast tumor growth and metastasis associated with decreased heparanase expression and Akt phosphorylation

OBJECTIVE GubenyiliuⅡ(GYⅡ),a traditional Chinese medicine(TCM)formula used in our hospital,has shown beneficial effects in cancer patients.In this study,we investigated the molecular mechanisms underlying the beneficial effects of GYⅡon murine breast cancer models.METHODS Inhibition of tumor growth and metastasis was evaluated by assessment of tumor weight and analysis of bioluminescent signal after a homograft inoculation.Viability of cultured breast cancer cells was determined using MTT assay andreal-time cell analysis(RTCA).Cell migratory ability was evaluated by Transwell?assay and wound healing assay.Subsequently,the potential anti-tumor and anti-metastatic mechanism was investigated by Western blotting and Immunohistochemistry.RESULTS GYⅡshowed significant inhibitory effects on tumor growth and metastasis in the murine breast cancer model.And GYⅡsuppressed theproliferation of 4T1 and MCF-7 cells in a dose-dependent manner.A better inhibitory effect on 4T1 cells proliferation and migration was found in sub-fractions(SF)of GYⅡ.Moreover,heparanase expression and degree of angiogenesis were reduced in tumor tissues.Western blotting analysis showed decreased expression of heparanase and growth factors in the cells treated with GYⅡand its sub-fractions(SF2 and SF3),there by a reduction in phosphorylation of ERK and AKT.CONCLUSION GYⅡexerts anti-tumor growth and anti-metastatic effects on murine breast cancer model.Sub-fractions 2 and 3 exhibits higher potency of the anti-tumor activity that is,at least partly,associated with decreased heparanase and growth factor sexpression,which subsequently sup-pressed activation of ERK and AKT pathways.

纳米银敷料联合人表皮生长外溶因子凝胶制剂在深Ⅱ度烧伤创面治疗中的应用效果

目的:探讨纳米银敷料联合人表皮生长外溶因子凝胶制剂在深Ⅱ度烧伤创面治疗中的应用效果。方法:选取汕头市第二人民医院收治的74例深Ⅱ度烧伤患者作为研究对象,以随机数表法分为对照组与观察组,每组各37例。全部患者均实施彻底清创,对照组在清创后采用人表皮生长外溶因子凝胶制剂治疗,观察组在对照组基础上加用纳米银敷料治疗,比较两组患者治疗效果。结果:相较于对照组,观察组治疗后视觉模拟评分法(VAS)评分与创面细菌阳性率更低,差异有统计学意义(P<0.05);观察组治疗1周与2周后的创面溶痂率更高,差异有统计学意义(t=11.501、10.102,P<0.05);观察组完全溶痂时间与创面愈合时间更短,差异有统计学意义(t=7.481、5.990,P<0.05);观察组治疗2周后的降钙素原(PCT)与高敏C反应蛋白(hs-CRP)均低于对照组,差异有统计学意义(t=6.259、12.689,P<0.05)。结论:纳米银敷料联合人表皮生长外溶因子凝胶制剂在深Ⅱ度烧伤创面治疗中的应用效果确切,可缩短创面愈合时间。

高亲和PDGFβR多肽修饰的截短型TβRⅡ的可溶性表达条件优化

目的 探讨高亲和血小板衍生生长因子受体(Platelet derived growth factor-β receptor, PDGFβR)多肽ZPDGFβR修饰的截短型转化生长因子β Ⅱ型受体(Truncated transforming growth factor-β receptor type Ⅱ,tTβRⅡ)的重组蛋白(Z-tTβRⅡ)最佳可溶性表达的条件。方法 将含有原核表达质粒pET28/Z-tTβRⅡ的阳性大肠杆菌(Rossta/pET28-Z-tTβRⅡ)置于LB培养基中培养,接着加入不同浓度的IPTG,使其终浓度为0.2、0.4、0.6、0.8 mmo/L,应用SDS-PAGE技术分析在不同温度时间下(16℃、18 h, 20℃、20 h和37℃、6 h)重组蛋白Z-tTβRⅡ的可溶性表达。结果 Z-tTβRⅡ在16℃、18 h下,0.8 mmol/L IPTG时,诱导后全菌表达约22%和诱导后上清表达约13%;Z-tTβRⅡ在20℃、20 h下,诱导后全菌0.8 mmol/L IPTG时表达约23%,诱导后上清0.2 mmol/L IPTG时表达约14%;Z-tTβRⅡ在37℃、6 h下,0.2 mmol/L IPTG时,诱导后全菌约30%和诱导后上清约20%。结论 Z-tTβRⅡ蛋白最佳表达条件为37℃、6 h、IPTG浓度为0.2 mmo/L。

IGF-Ⅱ和IGF-Ⅰ R在大肠癌中的表达

目的研究胰岛素样生长因子-(IGF-)和胰岛素样生长因子受体(IGF-R)在国人大肠癌中的表达情况及其与临床病理的关系。方法应用免疫组织化学过氧化物酶标记链霉卵白素法(S-P)对40例大肠癌患者的大肠癌组织、距大肠癌原发灶1cm癌旁组织、距大肠癌原发灶10cm以上大肠组织和10例非肿瘤患者的正常大肠组织标本,进行IGF-和IGF-R的抗原染色。结果1大肠癌组织和癌旁组织中IGF-和IGF-R的表达率显著高于切缘组织和正常大肠组织的表达率(P<0.05)。癌组织和癌旁组织中IGF-和IGF-R的表达率无显著性差异(P>0.05)。切缘组织和正常大肠组织中IGF-和IGF-R的表达率无显著性差异(P>0.05)。2在有淋巴结转移和Dukes分期C期及浸透全层组中IGF-和IGF-R的表达率分别显著高于无淋巴结转移组和Dukes分期A、B期及未浸透全层组中GF-和GF-R的表达率(P<0.05)。结论IGF-、IGF-R的高表达与大肠癌的发生发展和侵袭转移密切相关。

骨性关节炎软骨细胞中胰岛素样生长因子Ⅱ表达的研究

目的研究骨性关节炎(OA)软骨组织中细胞因子-胰岛素样生长因子Ⅱ(IGF-Ⅱ)的基因表达的变化规律。方法选取OA患者(6例)和创伤性截肢的患者(5例)的膝关节软骨组织,HE染色光镜观察OA软骨组织病理学特征。采用原位杂交方法定位软骨组织中IGF-ⅡmRNA的表达。提取关节软骨组织的总RNA和蛋白质,RT-PCR和Western印迹杂交方法测定软骨组织中IGF-ⅡmRNA和蛋白质的表达。结果OA软骨组织表面粗糙不平,软骨细胞排列紊乱,数目减少,有细胞成簇现象,可见软骨细胞变性、坏死,软骨厚度变薄,基质染色不均匀,有异染性。正常软骨细胞的细胞核与细胞浆中有IGF-ⅡmRNA的表达。与正常成人相比,OA患者软骨组织内IGF-Ⅱ在基因的转录与翻译水平明显降低了75%和63%(P<0.05)。结论细胞因子IGF-Ⅱ在OA软骨细胞中呈低表达状态。IGF-Ⅱ基因的差异表达可能是骨关节软骨退变的发生机制之一。