Expression of reg protein in rat regenerating islets and its co-localization with insulin in the Beta cell secretory granules

Summary Regenerating islets can be induced by the administration of poly(ADP-ribose) synthetase inhibitors to 90% depancreatized rats. In screening a regenerating isletderived cDNA library, we previously isolated a novel gene, reg (regenerating gene), which encodes a 165-amino acid protein with a 21-amino acid signal sequence. In the present study, we have examined the expression and localization of reg protein in the regenerating islets by immunocytochemical techniques using a monoclonal antibody against a recombinant rat reg protein of 144 amino acids without the signal sequence. Light microscopy examination showed strong immunoreactivity for reg protein in the regenerating islets of the rats at two weeks and two months after the 90% pancreatectomy, whereas reg protein was almost undetectable in normal rat islets or in the islets of the rats one year after the pancreatectomy. Almost all the reg protein-positive cells were stained for insulin. By applying the immunogold technique at the ultrastructural level, it was demonstrated that both reg protein and insulin occur in the central granular core of the regenerating Beta cell secretory granules. These results suggest that reg protein is synthesized in and secreted from the regenerating Beta cells and that its expression is closely associated with Beta-cell regeneration.     

葛花醒酒养胃颗粒对酒精性肝病内毒素损伤的防治作用

目的探讨葛花醒酒养胃颗粒对酒精性肝病(ALD)内毒素损伤的防治作用.方法Wistar大鼠48只随机分为正常组、模型组、葛花醒酒养胃颗粒治疗组(简称治疗组)和肝泰乐对照组(简称对照组).实验采用梯度酒精定量灌胃法制备酒精性肝病大鼠模型,造模同时治疗组予葛花醒酒养胃颗粒汤150mg·kg-1·d-1,分2次灌胃,对照组予肝泰乐水38mg·kg-1·d-1,分2次灌胃,正常组予同容积生理盐水,每日灌胃2次,连续灌胃8周后采血,检测大鼠血浆内毒素(LPS)、血清酶(ALT、AST、GGT)、血脂(Tch、TG)及血清肿瘤坏死因子(TNF-α)的含量,同时用RT-PCR法测定肝组织中脂多糖结合蛋白(LBP)和脂多糖受体CD14mRNA的表达.结果模型组大鼠血浆内毒素和血清TNF-α水平明显高于对照组(P＜0.05),肝组织中LBP和CD14mRNA的表达明显高于对照组(P＜0.05);同时血清ALT、AST、GGT、TG明显升高(P＜0.05);治疗组血浆内毒素和血清TNF-α水平明显低于模型组(P＜0.05),LBP和CD14mRNA的表达明显低于对照组(P＜0.05),血清ALT、AST、GGT、TG水平明显降低(P＜0.05).结论葛花醒酒养胃颗粒对大鼠酒精性肝损伤有较好的防治作用.     

灵葜四君颗粒联合化疗治疗胃癌的临床研究

目的:观察灵葜四君颗粒联合化疗治疗胃癌的临床疗效.方法:80例胃癌术后患者随机分为治疗组(灵葜四君颗粒加化疗)和对照组(单纯化疗)各40例.观察治疗前后卡氏评分、外周血常规和血液流变学的变化,并观察自然杀伤(NK)细胞、T淋巴细胞亚群、可溶性白细胞介素-2受体(SIL-2R)等指标的变化.结果:治疗组化疗后生存质量改善,外周血白细胞和红细胞数稳定;对照组外周血白细胞、红细胞数、血红蛋白浓度均较化疗前明显下降.治疗组化疗后NK细胞活性、CD3、CD4阳性T细胞数和CD4/CD8比值升高,血清SIL-2R的浓度和全血粘度(低切、中切、高切)均明显下降;而对照组化疗后NK细胞活性、CD4阳性T细胞数和CD4/CD8比值下降,血清SIL-2R的浓度升高而全血粘度无明显改变.结论:灵葜四君颗粒对化疗有增效减毒作用,对胃癌术后患者机体免疫功能有保护和促进作用,并能防治化疗引起的骨髓抑制,降低血液粘度.     

Anti-eosinophil peroxidase antibodies detected in patients with primary biliary cirrhosis.

OBJECTIVE:: The possible involvement of eosinophils in primary biliary cirrhosis (PBC) has been suggested for many years. Activated eosinophils release basic granule proteins and are thought to be involved in the tissue damage of PBC. Eosinophil peroxidase (EPO) is one of the eosinophilic granule proteins and is toxic to epithelial cells. In this study, we examined whether autoantibodies to EPO are present in sera of patients with PBC and we evaluated the relationship between the presence of anti-EPO antibodies and the clinical features of PBC. METHODS:: Sera obtained from 61 patients with PBC, 31 patients with autoimmune hepatitis (AIH), 87 patients with chronic viral hepatitis (CVH), 27 patients with bronchial asthma, and 20 healthy controls were examined. Enzyme-linked immunosorbent assay (ELISA) and Western blot methods were used for detection of anti-EPO antibodies. RESULTS:: The mean OD value of anti-EPO antibodies, as determined by ELISA, was significantly (p<0.01) higher in patients with PBC (0.197+/-0.121) than in patients with AIH (0.124+/-0.077), CVH (0.090+/-0.038) or bronchial asthma (0.073+/-0.025) and in healthy controls (0.072+/-0.029). The results of ELISA showed that 32 (52.5%) of the 61 patients with PBC, 9 (29.0%) of the 31 patients with AIH, 7 (8.0%) of the 87 patients with CVH, and 1 (3.7%) of the 27 patients with bronchial asthma were positive for anti-EPO antibodies. In addition, PBC patients who were positive for anti-EPO antibodies had a significantly smaller number of peripheral eosinophils than did patients who were negative for anti-EPO antibodies (99.2+/-54.4cells/mul versus 176.9+/-117.5cells/mul, p<0.01). However, there was no correlation between the titers of anti-EPO antibodies and those of AMA or AMA-M2, ANA or serum levels of IgM. CONCLUSIONS:: This is the first report of the detection of anti-EPO antibodies in patients with PBC. Further study is needed to clarify the role of anti-EPO antibodies in the pathogenesis of PBC.     

血浆D二聚体和GMP-140测定在DIC诊断中的意义

目的：探讨分子标志物在弥散性血管内凝血（DIC）诊断中的价值。方法：分别检测DIC患者20例（DIC组）、DIC疑诊组11例（DIC疑诊者）和正常人10例（正常对照组）血浆D二聚体和血小板颗粒膜蛋白（GMP－140）水平。结果：以上三组血浆D二聚体水平分别为4．43±3．43，2．03±0．17，0．36±0．17（mg／L）。血浆GMP－140水平分别为53．78±27．55，33．32±16．50，17．29±4．84（ng／ml）。DIC组血浆D二聚体和GMP－140显著高于DIC疑诊组和正常对照组，而DIC疑诊组与正常对照组差异尤显著性。动态观察发现，血浆D二聚体升高的6例DIC疑诊者有3例随后转变成DIC。结论：血浆D二聚体和GMP－140是DIC时敏感指标，监测二指标有助于DIC的早期诊断。     

养胃颗粒中NF-κB靶标介导的治疗胃溃疡成分的筛选与评价研究

目的筛选养胃颗粒中影响NF-κB靶标治疗胃溃疡的化学成分,并建立基于上述成分的质量评价方法。方法采用NF-κB质粒转染的RAW264.7细胞株,给药、孵育后使用PCR进行扩增和表达,测定荧光值以评价各化学成分对NF-κB靶标的影响;再以筛选所得成分为指标,采用LC-MS法进行多指标含量测定研究,色谱柱为Agilent SB C18(100 mm×2.1 mm,2.7μm),使用0.1%甲酸-乙腈作为流动相,梯度洗脱,ESI模式同时进行正负离子扫描,离子对为毛蕊异黄酮葡萄糖苷786/425,三七皂苷R1为945/925,共测定10批养胃颗粒。结果养胃颗粒的7个成分中,毛蕊异黄酮葡萄糖苷和三七皂苷R1具有较好的NF-κB靶标激动活性,在10批颗粒中,毛蕊异黄酮葡萄糖苷的平均含量为0.834 mg/g,三七皂苷R1的平均含量为0.512 mg/g。结论养胃颗粒中,毛蕊异黄酮葡萄糖苷、三七皂苷R1是通过NF-κB靶标治疗胃溃疡的活性成分,本研究以上述成分为指标建立LC-MS质量评价方法,具有准确、高效、贴近功效主治的特点。     

小儿伤科泡腾颗粒对闭合性骨折模型大鼠治疗效果研究

目的对比分析小儿伤科汤剂与小儿伤科泡腾颗粒对闭合性骨折模型大鼠的治疗效果。方法选取45只幼年雄性SD大鼠作为研究对象,利用骨折器建立大鼠胫骨闭合性骨折模型。根据随机数字表原则,将模型大鼠随机分为对照组、汤剂组和颗粒组,每组15只。所有大鼠手法复位固定后,对照组大鼠采用温开水灌胃作为对照,汤剂组大鼠采用小儿伤科汤剂灌胃治疗,颗粒组大鼠采用小儿伤科泡腾颗粒灌胃治疗。分别在治疗14 d、28 d、35 d时,每组处死5只大鼠,采集腹主动脉血检测血清钙、血清磷、血清碱性磷酸酶(ALP)指标,同时对患肢进行X摄片检查骨折愈合情况,以Lane-Sandhu X射线评分标准评价骨缺损修复情况。结果治疗14 d时,三组大鼠血清钙、血清磷、血清ALP指标比较差异均无统计学意义(P>0.05)。治疗35 d时,汤剂组和颗粒组大鼠血清钙指标低于对照组,差异有统计学意义(P<0.05)。但汤剂组和颗粒组大鼠比较差异无统计学意义(P>0.05)。治疗28 d、35 d时,汤剂组和颗粒组大鼠血清ALP高于对照组,差异有统计学意义(P<0.05)。但汤剂组和颗粒组大鼠比较差异无统计学意义(P>0.05)。治疗14 d时,三组大鼠Lane-Sandhu评分比较差异无统计学意义(P>0.05)。治疗28 d、35 d时,汤剂组和颗粒组大鼠Lane-Sandhu评分均高于对照组,差异有统计学意义(P<0.05)。但汤剂组和颗粒组大鼠Lane-Sandhu评分比较差异无统计学意义(P>0.05)。结论小儿伤科汤剂与小儿伤科泡腾颗粒治疗闭合性骨折模型大鼠效果相当。     

麦芽总生物碱颗粒剂的制备

目的:在前期药效实验的基础上,提取麦芽总生物碱,优选麦芽总生物碱颗粒剂成型制备工艺,得到稳定均一,质量合格的颗粒制剂。方法:利用超声提取法,提取麦芽总生物碱;采用酸性染料比色法测定麦芽中总生物碱的含量;拟选6个处方完成颗粒剂的制备,并以休止角、成型率、堆密度和吸湿性等为指标,应用综合评分法对各处方进行打分,筛选出最优的辅料处方,完成辅料用量比例的优化,并筛选出最佳润湿剂;对制备所得到的麦芽总生物碱颗粒的溶化性,粒度及干燥失重等进行测定。结果:以糊精和乳糖比例为1:1的混合物作为辅料,并按1:1的比例混合麦芽总生物碱浸膏粉与辅料,选取80%的乙醇作为润湿剂,制备所得颗粒剂大小均一,成型率最高。结论:本文成功优选出了麦芽总生物碱颗粒剂的制备工艺,制成的颗粒溶化性好,成型率佳,制备方法稳定可靠。     

胃肠舒泰颗粒对鼠血胃动素及胃肠运动的影响

[目的]观察胃肠舒泰(WCST)颗粒对小鼠胃排空及小肠推进作用和大鼠血清胃动素(MOT)的影响.[方法]ICR小鼠100只,胃排空实验、小肠推进实验各选50只,分别随机分为5组,空白对照组、WCST颗粒小、中、大剂量组及西沙比利组,观察各组胃排空及小肠推进率.SD大鼠50只,亦分5组,观察WCST颗粒对大鼠血清MOT的影响.[结果]WCST颗粒大、中剂量组小鼠胃排空率及小肠推进率较空白对照组均明显增快(P＜0.05～＜0.01),大、中剂量组大鼠血清中MOT亦明显增高(P＜0.05～0.01).[结论]WCST颗粒具有促进小鼠胃排空及小肠推进率和升高大鼠血清MOT的作用.这一机制可能与促进内源性MOT释放有关.