胃泌素释放肽及其受体在结直肠癌中的表达

目的研究胃泌素释放肽（GRP）及胃泌素释放肽受体（GRPR）在结直肠癌中的表达。方法采用免疫组化方法测定结直肠癌中GRP和GRPR的表达。结果 GRP和GRPR在结直肠癌中阳性表达,阳性表达率分别为65.3%和79.6%,随着病情严重程度增加表达增加。结论 GRP和GRPR的异常表达与结直肠癌有关,可作为结直肠癌治疗的目标。     

胃泌素释放肽及其受体在结直肠癌中的表达

目的探讨结直肠癌中胃泌素释放肽与预后、远处转移的关系,同时分析其在远处转移中的预测价值。方法收集我院62例诊断为结直肠癌患者的临床及病理资料,采用前瞻性研究方法,利用免疫组化技术了解患者中胃泌素释放肽(GRP)、胃泌素释放肽受体(GRPR)蛋白的表达,结合肿瘤体积进行分析,了解二者与结直肠癌患者预后、远处转移的关系。结果 GRP、GRPR蛋白在结直肠癌组织中阳性表达率分别为79.0%和83.2%,二者表达存在相关性。GRP、GRPR蛋白水平与结直肠癌患者的预后及远处转移差异具有一定的相关性。GRP、GRPR能够在一定程度上预测结直肠癌远处转移,敏感度及特异度为69.4%、71.3%及70.1%、65.5%。结论 GRP、GRPR能够较好的预测结直肠癌患者的预后及远处转移,具有较好的敏感性及特异性。     

儿童空腹血浆胃泌素和胃泌素释放肽的水平

应用放射免疫法测定了125例儿童(年龄自2月至14岁)空腹血浆胃泌素(gastrin)和胃泌素释放肽(gastrin-releasing peptide,GRP)。胃泌素和GRP的均值和实测范围分别为14.0±14.9,0～98.0 pmol/L和8.8士8.2,0～48.0 pmol/L。两种激素的浓度无显著的性别差异。胃泌素以2月至3岁以下组的含量为最高,其均值为12～14岁组的4倍以上。结果表明,胃泌素的浓度与年龄呈负相关(r=-0.50,P<0.001),而GRP的浓度则与年龄无相关性(P>0.05)。本文讨论了在不同年龄组的儿童的空腹血浆浓度的变化。     

胃泌素释放肽前体对小细胞肺癌诊治的临床意义

目的探讨血清胃泌素释放肽前体(ProGRP)的生物学特性及其对小细胞肺癌(SCLC)的诊断、病情监测、疗效评估、预后判断的临床意义以及ProGRP与神经元特异性烯醇化酶(NSE)水平之间的相关性,为SCLC的预防和治疗提供科学依据。方法收集肺癌患者1221例,健康对照组500名,采用酶联免疫吸附法检测治疗前后肺癌患者及健康对照者血清ProGRP和NSE水平,同时肺癌患者还进行相关的影像学检查。结果270例治疗前SCLC患者血清ProGRP中位值为544.48pg/ml,敏感性为84.07%,均明显高于非小细胞肺癌(NSCLC)组及健康对照组,差异均有统计学意义(P<0.01)。270例SCLC患者随着临床分期期别的增加,Pro-GRP水平大于界值例数也明显增加,检出敏感性增加(P<0.01)。270例SCLC患者疗效评价获得完全缓解者ProGRP水平明显下降,无变化者及病情进展者ProGRP水平均明显高于界值(P<0.01)。治疗后在24个月之内出现复发、转移患者,血ProGRP水平明显高于治疗后健康生存者,且高于治疗前血ProGRP水平(P<0.01)。ProGRP水平的变化与影像学所示的肿瘤大小具有明显的相关性,ProGRP和NSE水平对SCLC的联合检测呈正相关(r=0.243,P<0.01)。结论血清ProGRP检测可作为SCLC诊断、病情监测、疗效评估、预后判断的敏感和特异性指标,与NSE联合检测可进一步提高敏感性。     

抗胃泌素释放肽单克隆抗体的制备及其在体外对乳腺癌细胞生长的抑制

研制抗胃泌素释放肽(Gastrin-releasing peptide,GRP)单克隆抗体,并且初步评估其在体外对乳腺癌细胞生长的抑制作用。纯化重组的GRP融合蛋白,免疫BALB/c小鼠,取经免疫的小鼠脾细胞与小鼠骨髓细胞SP2/0进行细胞融合,对杂交瘤细胞进行筛选,阳性孔经3次有限稀释法亚克隆,获得稳定分泌GRP抗体的杂交瘤细胞株,进一步从腹水中用硫酸铵沉淀法纯化单抗,并以间接ELISA、Western免疫印迹以及亚类分型等方法进行抗体鉴定,之后在体外用MTT比色法测定该单抗抑制乳腺癌细胞EMT-6生长的作用。结果表明通过筛选获得了一株阳性杂交瘤细胞,命名为W9,这株杂交瘤细胞分泌的单抗具有效价高,特异性好的特点,其亚型为IgG1,体外的细胞实验也表明这种单克隆抗体能够显著地抑制EMT-6细胞的增殖。     

胃泌素释放肽前体在小细胞肺癌诊断及治疗中应用探讨

目的:探究胃泌素释放肽前体在小细胞肺癌诊断及治疗中应用.方法:选取2018年12月～2019年11月某院收治的小细胞肺癌和非小细胞肺癌患者各65例,分为小细胞肺癌患者组和非小细胞肺癌患者组.结果:小细胞肺癌组的ProGRP(胃泌素释放肽前体)、CEA(癌胚抗原)以及NSE(神经元特异度烯醇化酶)等血清指标高于非小细胞肺...     

胃泌素释放肽前体检测对小细胞肺癌诊治的意义

目的探讨血浆胃泌素释放肽前体(ProGRP)对小细胞肺癌(SCLC)的临床诊断价值。方法用化学发光法检测50例健康体检者、49例非小细胞肺癌(NSCLC)及50例初诊SCLC(局限期29例、广泛期21例)患者血浆ProGRP水平,评估血浆ProGRP诊断SCLC的临床价值。结果健康对照组、NSCLC组和SCLC组血浆ProGRP水平分别为34.3、39.1和403.1 ng/L;各组间血浆ProGRP差异均有统计学意义。局限期SCLC(LD-SCLC)组血浆ProGRP高于健康对照组,差别有统计学意义。广泛期SCLC(ED-SCLC)组血浆ProGRP高于LD-SCLC组,差别有统计学意义。以健康组为对照,ROC曲线上取约登指数最大点确定ProGRP临界值为36.9 ng/L,ProGRP的敏感度为88%,特异度为98%;以NSCLC组为对照,ROC曲线上取约登指数最大点确定ProGRP的临界值为62.9 ng/L,ProGRP的敏感度为82%,特异度为96%。结论血浆ProGRP检测可作为SCLC诊断、病情监测的敏感和特异性指标,可广泛应用于临床。     

胃泌素释放肽DNA疫苗抑制EMT6乳腺癌生长的实验研究

目的：观察胃泌素释放肽（GRP）DNA疫苗对EMT6小鼠乳腺癌生长的抑制作用。方法：将构建的GRP DNA疫苗pCR3.1-VS-HSP65-TP-GRP6-M2肌肉注射免疫BALB/c雌性小鼠,2周1次,共5次。采用ELISA法对小鼠的血清中抗GRP-IgG类抗体进行检测。最后一次免疫后第2周,接种EMT6小鼠乳腺癌细胞。于肿瘤细胞接种后d14,处死全部动物,称量肿瘤的质量和测量肿瘤的体积。并对瘤组织进行常规HE染色。结果：pCR3.1-VS-HSP65-TP-GRP6-M2免疫BALB/c小鼠,可诱导抗GRP-IgG类抗体的产生。并对随后的EMT6肿瘤细胞攻击有显著的抑制作用（P〈0.01）,抑瘤率为46.53%。病理学检查结果显示,GRP DNA疫苗成功地激发了宿主的抗肿瘤免疫反应;与pCR3.1-VS-HSP65-TP对照组相比,GRP DNA疫苗免疫组小鼠EMT6肿瘤组织浸润性下降。结论：GRP DNA疫苗显著抑制EMT6乳腺癌生长,为此类疫苗应用研究奠定了基础。     

Nonpeptide gastrin releasing peptide receptor antagonists inhibit the proliferation of lung cancer cells

The ability of nonpeptide antagonists to interact with gastrin releasing peptide receptors on lung cancer cells was investigated. PD176252 (3-(1H-Indol-3-yl)-N-[1-(5-methoxy-pyridin-2-yl)-cyclohexylmethyl]-2-methyl-2-[3-(4-nitro-phenyl)-ureido]-propionamide) and PD168368 (3-(1H-Indol-3-yl)-2-methyl-2-[3(4-nitro-phenyl)-ureido]-N-(1-pyridin-2-yl-cyclohexylmethyl)-propionamide) inhibited specific 125I-gastrin releasing peptide binding to NCI-H1299 cells with IC50 values of 20 and 1500 nM, respectively. Similar binding results were obtained using NCI-H157, H345 and N592 human lung cancer cells. PD176252 inhibited the ability of 1 nM bombesin to cause elevation of cytosolic calcium in Fura-2 loaded NCI-H345 or H1299 cells, whereas it had no effect on basal cytosolic calcium. PD176252 antagonized the ability of 10 nM bombesin to cause elevation of c-fos mRNA in NCI-H1299 cells. Also, PD176252 inhibited the ability of 100 nM bombesin to cause tyrosine phosphorylation of focal adhesion kinase in NCI-H1299 cells. Using a [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] assay, PD176252 was more potent than PD168368 at inhibiting NCI-H1299 proliferation. Also, 1 microM PD176252 significantly inhibited lung cancer colony number in vitro. PD176252 in a dose-dependent manner inhibited NCI-H1299 xenograft growth in nude mice in vivo. These results indicate that PD176252 is a gastrin releasing peptide receptor antagonist, which inhibits the proliferation of lung cancer cells.     

Synthesis of a tritiated human growth hormone releasing peptide

Tritium-labeled growth hormone releasing peptide His-D-Trp-Ah-Trp-D-Phe-Lys-NH₂ was synthesized by tritium-halogen exchange on the precursor His-5,7-Br₂-D-Trp-Ala-Trp-D-Phe-Lys-NH₂, The radiolabeled peptide had a specific activity of 29 Ci/mmol and a radiochemical purity of 95 %. The tritium label was shown by ³H NMR to be located mostly at the expected 5, 7-positions of the indole nucleus in the D-Trp residue. The dibromopeptide was prepared by solid-phase peptide synthesis, employing racemic 5,7-Br₂-Trp as a building block and separation of the resulting epimeric mixture by HPLC. 5,7-Br₂-Trp was prepared by a five-step sequence beginning with 2,4-dibromoaniline, The use of anisole as an additive in the HF resin/peptide cleavage was rejected because anisolc was found to undergo electrophilic substitution of the dibromoindole nucleus; a modified HF deprotection/cleavage procedure was developed and used instead.     

Impact of dianionic and dicationic linkers on tumor uptake and biodistribution of [64Cu]Cu/NOTA peptide‐based gastrin‐releasing peptide receptors antagonists

In this study, we investigated for the first time the influence of 2‐aminoethyl‐piperazine‐1‐carboxylic acid (APCA) and amino‐hexanedioic‐1‐acid (AHDA) on tumor uptake and elimination kinetics of [⁶⁴Cu]‐radiolabeled gastrin releasing peptide receptors (GRPR) antagonists. Three GRPR antagonists containing the RM26 sequence were synthesized and conjugated with NOTA via different linkers (LK): polyethylene glycol (PEG–neutral), APCA (dicationic) or AHDA (dianionic). The NOTA‐LK‐RM26 peptides were radiolabeled with ⁶⁴Cu to assess their pharmacokinetic and positron emission tomography (PET) imaging properties using PC3 tumor‐bearing athymic nude mice. The inhibition constants (K_i) of the 3 ^natCu/NOTA‐LK‐RM26 peptides bearing PEG, dicationic and dianionic linkers were 0.98 ± 0.48 nM, 0.95 ± 0.21 nM, and 17.97 ± 2.79 nM, respectively. The [⁶⁴Cu] NOTA‐LK‐RM26 conjugates were prepared with labeling yields superior to 95% and specific activities of 67 to 77 TBq/mmol. The 3 radiopeptides were stable in vivo and showed GRPR‐specific uptake in pancreas with a very fast washout of this tissue observed for [⁶⁴Cu]‐NOTA‐AHDA‐RM26 peptide. Results from imaging studies displayed specific PC3 tumor uptake for both [⁶⁴Cu]‐NOTA‐APCA‐ and AHDA‐RM26, similar kidney elimination and fast liver washout. Considering their adequate imaging characteristics, [⁶⁴Cu]‐NOTA‐LK‐RM26 bearing APCA‐ and AHDA‐linkers are promising candidates for GRPR‐targeted PET imaging prostate cancer.     

Histamine releasing peptide (HRP) has proinflammatory effects and is present at sites of inflammation

Albumin, the most abundant plasma protein, readily enters sites of inflammation during the period of increased vascular permeability. There it encounters proteases released from mast cells and invading leukocytes which earlier work has shown can act on albumin to liberate the peptide, histamine releasing peptide (HRP), first identified and named by its ability to stimulate histamine release from isolated mast cells. In this report we show that HRP releases histamine from cutaneous mast cells in vivo resulting in increased vascular permeability and persistent edema while in vitro, HRP promotes chemotaxis of leukocytes and enhances macrophage phagocytosis. Moreover, we show that the level of HRP is increased with the induction of an acute cutaneous inflammatory response in rats, that HRP is present at sites of acute and chronic inflammation in humans and that HRP is rapidly degraded by proteases thereby limiting its action to the area of its generation. We suggest that HRP is a pro-inflammatory peptide that helps amplify and perpetuate the inflammatory response. Inhibitors of inflammatory proteases or antagonists that block the action of peptides like HRP may, therefore, be useful in breaking the cycle of inflammation.     

Glucagon releasing activity of a cyclic peptide related to somatostatin

A possible benefit of creating smaller and more rigid active analogs of somatostatin is the discovery of compounds which selectively inhibit the secretion of insulin, glucagon or growth hormone. A series of cyclic tetrapeptide analogs related to somatostatin was synthesized, and one member of this series was found to cause an unexpected stimulation of glucagon secretion while having little if any effect on either insulin or growth hormone secretion. A sustained increase in plasma glucose levels was also observed. Two possible modes of action are proposed.     

IL-10基因克隆化与原核重组IL-10蛋白制备的实验研究

目的克隆化人IL-10基因及重组蛋白的制备为IL-10基因表达在小儿炎症性、过敏性疾病中的作用及治疗等提供有力的证据。方法采用过敏症患者发作时期的外周血,提取总RNA。以总RNA为模板,特异性引物进行反转录,RT-PCR进行IL-10基因筛选。PCR产物经酶切分析和DNA序列测定后,构建重组质粒pTrchHis2B-hIL10原核高效表达载体,通过大肠杆菌(工程菌)JM109经IPTG诱导表达重组hIL-10蛋白,并经与Ni-NTA树脂中的6×His配体结合进行亲和层析纯化,表达产物进行SDS-PAGE电泳分析。结果DNA测序显示克隆化人IL-10全长开放读框准确,表达读码框正确。SDS-PAGE分析显示,原核表达系统表达的重组蛋白量和纯度满意。结论采用以总RNA为模板的RT-PCR一步法,能简便、可靠地获得hIL-10基因产物,所构建的原核表达载体pTrcHis2B-hIL10,能够快速、高效制备rhIL-10蛋白,为细胞因子hIL-10功能的进一步研究及临床应用奠定了基础。     

冻干重组人脑利钠肽在老年顽固性心力衰竭患者中的临床应用及护理干预

目的观察冻干重组人脑利钠肽治疗老年顽固性心力衰竭的临床疗效及护理干预的作用。方法选取21例确诊为顽固性心力衰竭的老年患者,在标准化治疗基础上加用冻干重组人脑利钠肽治疗,负荷剂量1．5μg/kg静脉推注,随后按维持剂量0．0075μg/（kg·min）进行静脉滴注,持续24～72h。采用治疗前后自身对照,观察用药前后血压、心率、尿量、呼吸困难程度、下肢水肿情况、纽约心脏病协会（NYHA）心功能是否改善以及血B型脑利钠肽（BNP）浓度,中心静脉压（CVP）监测的变化。并对患者进行护理干预。结果与治疗前比较,治疗后患者血压、心率、血BNP及CVP测定值均明显下降[（128±15）mmHg（1mmHg=0．133kPa）比（144±17）mmHg,（84±9）次／min比（102±13）次／min,（560±180）μg／L比（14501±430）μg／L,（9．2±2．3）mmHg比（14．9±3．6）mmHg],差异均有统计学意义（均P〈0．05）。治疗后患者心功能Ⅱ级12例,Ⅲ级6例,Ⅳ级3例。治疗后显效7例,有效11例,无效2例,死亡1例。治疗前后NYHA分级比较,差异有统计学意义（P〈0．05）。结论顽固性心力衰竭患者病情进行性加重,冻干重组人脑利钠肽治疗有效,精心的护理对患者康复至关重要。     

重组人脑利钠肽安全药理学研究

目的:观察重组人脑利钠肽(rhBNP)对大鼠心血管、呼吸系统和对小鼠神经系统的影响。方法:大鼠实验分低、中、高3个剂量及空白对照共4个组,给药组分别单次静脉注射(iv)rhBNP 22．5,45．0和90．0μg·kg~(-1),对血压、心电图(ECG)、呼吸频率及节律进行观察。小鼠实验分低、中、高3个剂量及空白对照共4个组,给药组分别单次iv rhBNP 45．0,90．0和180．0μg·kg~(-1),对动物自发活动和爬杆能力进行观察。结果:rhBNP使大鼠收缩压、舒张压、平均动脉压降低,且存在量效关系,2h内上述指标基本恢复正常,对心率、心律、ECG之QRS时间、T波、ST段、呼吸频率、节律和幅度无明显影响。对小鼠自发活动和爬杆能力无明显影响。结论:rhBNP对心血管系统有一定影响,可使收缩压、舒张压、平均动脉压降低。     

重组人脑钠素对恒河猴长期毒性研究

目的:研究重组人脑钠素(rhBNP)对恒河猴的长期毒性.方法:健康恒河猴分别按体重随机分为低、中、高剂量组和空白对照组.低、中、高剂量组给药量分别为0.03,0.09和0.3mg·kg-1.连续30d静滴给药.末次给药后处死一半动物做病理解剖,另一半停药后继续观察15d.观察症状和检测指标包括:①血压、尿量等一般症状.②心电图.③红细胞、白细胞等血液学指标.④血钾、钠、氯、醛固酮等血液生化指标.⑤尿液检查.⑥抗体测定.⑦骨髓检查.⑧注射部位、心脏等病理检查.结果:动物连续给药后,高剂量组猴静滴后血压明显下降,中剂量组给药后血压有下降趋势但无明显差异.血压变化在3h内恢复正常.高、中剂量组给药后2h内尿量明显增加,但24h总尿量无明显改变.低剂量组血压及尿量无明显变化.d30病理组织学检查发现高剂量组少数动物出现肝细胞坏死,d45未发现有上述病理变化.肾组织也发现有病理变化,但存在于各个组,可能非药物所致.结论:rhBNP对猴心血管、泌尿系统有一定的药理毒理作用,主要表现为降血压、利尿,对肝脏可能有轻度毒性作用,其作用均是可逆的.rhBNP对恒河猴的安全剂量为0.03mg·kg-1.     

Applications of cyclic peptide nanotubes (cPNTs)

Self-assembled cyclic peptide nanotubes (cPNTs) have recently drawn particular attention as one of the most intriguing nanostructures in the field of nanotechnology. Given their unique features including high surface area, increased drug loading, environmental stability, enhanced permeation, and modifiable drug release, these hollow tubular structures can be constructed with cyclic di-, tri-, tetra-, hexa-, octa-, and decapeptides with various amino acid sequences, enantiomers, and functionalized side chains and can be applied for antiviral and antibacterial drugs, drug delivery and gene delivery vectors, organic electronic devices, and ionic or molecular channels. Recent publications have presented promising results regarding the use of cPNTs as drugs or biomedical devices. However, there is an urgent need for the further in vivo nanotoxicity and safety testing of these nanotubes to evaluate their suitability in different fields.