DEAD盒多肽43小干扰RNA对司美替尼抑制人肺腺癌A549细胞增殖和诱导其凋亡作用的影响

目的探讨DEAD盒多肽43(DDX43)小干扰RNA(siRNA)对司美替尼抑制人肺腺癌A549细胞增殖和诱导其凋亡作用的影响。方法培养A549细胞,并分为对照组、司美替尼组和司美替尼+DDX43 siRNA组,运用细胞免疫组化和原位杂交的方法分别检测各组A549细胞中DDX43蛋白和mRNA的表达情况,采用MTT法检测各组细胞的增殖情况,采用TUNEL法检测各组细胞的凋亡情况。结果对照组、司美替尼组、司美替尼+DDX43 siRNA组DDX43蛋白相对表达量分别为138.20±17.72、79.95±8.96、36.16±6.31,总体比较差异有统计学意义(F=66.72,P=0.02);表达水平依次降低,且两两比较差异均有统计学意义(P均<0.05)。对照组、司美替尼组、司美替尼+DDX43 siRNA组DDX43 mRNA相对表达量分别为266.20±15.07、164.95±8.96、71.16±6.31,总体比较差异有统计学意义(F=125.35,P=0.01);表达水平依次降低,且两两比较差异均有统计学意义(P均<0.05)。对照组、司美替尼组、司美替尼+DDX43 siRNA组A549细胞凋亡指数分别为(3.75±0.55)%、(11.72±1.06)%、(19.98±1.21)%,总体比较差异有统计学意义(F=12.31,P=0.04);凋亡指数依次升高,且两两比较差异均有统计学意义(P均<0.05)。对照组、司美替尼组、司美替尼+DDX43 siRNA组A549细胞生长抑制率分别为(2.11±0.13)%、(14.91±0.29)%、(27.14±0.58)%,总体比较差异有统计学意义(F=15.23,P=0.03);生长抑制率依次升高,且两两比较差异均有统计学意义(P均<0.05)。结论DDX43 siRNA联合司美替尼可抑制A549细胞增殖,促进其凋亡。     

乙型肝炎病毒preS1 多肽在大肠杆菌中的表达与纯化研究

目的　获得大量完整的重组乙肝病毒（ Ｈ Ｂ Ｖ）ｐｒｅ Ｓ１ 多肽纯品,以利于ｐｒｅ Ｓ１ 功能与免疫学性质的研究。方法　比较不同表达载体及不同的培养、诱导和纯化条件,最大限度地使表达产物免受大肠杆菌蛋白酶的降解。结果　分析不同长度ｐｒｅ Ｓ１ 基因在两种不同载体,两种表型宿主菌中表达产物的状况,可看出在ｐｒｅ Ｓ１ 第５６ 和６４ 位氨基酸之间存在两个 Ｅ．ｃｏｌｉ 蛋白酶切割点。用ｐ Ｑｅ９载体和 Ｍ１５（ｐ Ｒ Ｅ Ｐ４） 宿主菌, Ｉ Ｐ Ｔ Ｇ 诱导１２ 小时以 Ｎｉ Ｎ Ｔ Ａ 琼脂糖柱在变性条件下纯化,能获得１０ ｍｇ／ Ｌ 以上具有良好免疫学活性的电泳纯ｐｒｅ Ｓ１ 蛋白。结论　缩短诱导时间、用一步法在变性条件下纯化表达产物可获得无明显降解的ｐｒｅ Ｓ１ 完整蛋白。     

Electrocatalytic activity of a cobalt hexacyanoferrate modified glassy carbon electrode toward ascorbic acid oxidation

A cobalt hexacyanoferrate (CoHCF) modified glassy carbon (CoHCF/GC) electrode was prepared electrochemically. The voltammetric responses of CoHCF are stable and the electrochemical behaviour is related to the concentrations of supporting electrolyte and counterions. The CoHCF/GC electrode shows electrocatalytic activity toward the oxidation of ascorbic acid in phosphate buffer solution. The electrocatalytic rate constant of the CoHCF/GC electrode for the oxidation of ascorbic acid is determined using rotating disk electrode measurements.     

Colonic enteric nervous system in patients with familial amyloidotic neuropathy

The colonic enteric nervous system was investigated in autopsy specimens from 12 patients with familial amyloidotic neuropathy (FAP) and 9 controls. The infiltration of amyloid deposits in the enteric nervous system was studied by double staining for amyloid and nerve elements. The myenteric plexus was immunostained for protein gene product (PGP) 9.5, vasoactive intestinal peptide (VIP), substance P and nitric oxide synthase (NOS). The immunostained nerve elements were quantified by computerised image analysis. Double staining revealed that there was no amyloid infiltration in the ganglia, or in the nerve fibres in the colonic enteric nervous system of FAP patients. The relative volume density of PGP 9.5-immunoreactive nerve fibres in both the circular and the longitudinal muscle layers in FAP patients did not differ significantly from that of controls. The relative volume density of VIP-immunoreactive nerve fibres in the circular muscle layer was significantly decreased in FAP patients compared with controls, but not in the longitudinal layer. The number of VIP-immunoreactive neurons/mm² myenteric ganglia was significantly decreased in FAP patients. There were no statistical differences in the relative volume density for substance P- and NOS-immunoreactive nerve fibres between FAP patients and controls, nor was there any difference between FAP patients and controls regarding the number of NOS- and substance P-immunoreactive neurons/mm² myenteric ganglia. It is concluded that the colonic enteric nervous system as a whole is intact and is not damaged by amyloid infiltration. The present observation of a reduction of VIP-immunoreactive nerve fibres and neurons in myenteric plexus of FAP patients might be one of the factors that contribute to the motility disorders seen in FAP patients. Received: 1 September 1998 / Revised, accepted: 26 November 1998     

真实世界中鹿瓜多肽注射液对骨折人群肾功能的影响实效研究

目的:探究真实世界骨折人群患者使用鹿瓜多肽注射液是否会对肾功能造成影响.方法:在全国26家三甲医院的医院信息系统(HIS)中选取使用鹿瓜多肽注射液的骨折患者(鹿瓜组)12847例和未使用鹿瓜多肽注射液的骨折患者(对照组)12619例的信息进行研究.运用GBM倾向评分加权法控制混杂因素,通过Logistic对比分析真实世...     

Peptide containing nerves in the ureter of the guinea-pig and cat

We report the presence of two regulatory peptides, substance P and vasoactive intestinal polypeptide (VIP), in the ureter and their localisation by both light- and electron-microscopy to autonomic nerves. VIP- and substance P-like immunoreactive nerves showed, in general, a similar anatomical distribution in the various layers of the ureter. Immunoreactive nerves were observed running along the smooth muscle coat, parallel to muscle bundles, around blood vessels and in the submucosa, particularly beneath the epithelium. In addition, scattered VIP-like immunoreactive ganglion cells and nerve fibres were seen in adventitial ganglia around the most distal part of the ureter and ureter-bladder junction in the cat. The guinea-pig ureter contained principally substance P-like immunoreactivity, whereas the cat ureter possessed mainly VIP-like material.The distribution of adrenergic and cholinergic nerves was compared with those containing peptides. Peptide-containing nerves had a more extensive distribution than adrenergic ones, which were mainly associated with blood vessels; however, cholinergic nerves were often localised in the same areas as those possessing peptides. Conventional electron microscopy revealed that separate p-type (peptidergic) and cholinergic nerve terminals were frequently present in the same nerve bundles, although in the cat ureter some 50% of the p-type profiles contained a mixed population of vesicles, characteristic of both cholinergic and p-type nerves. VIP- and substance P-like immunoreactivity were also localised at the ultrastructural level by means of a gold-labelled goat-antirabbit serum.     

pCH510转染膀胱癌细胞系BIU-87对卡介苗抑瘤作用的影响

目的研究膀胱癌细胞系BIU-87体外转染pCH510后,CH50多肽的表达及其对卡介苗(bacilluscalmette-guerin,BCG)抑瘤作用的影响。方法在LipofectimineTM2000的介导下,将质粒pCH510体外转染给BIU-87细胞,采用免疫组织化学S-P法和WesternBlot法鉴定CH50多肽的表达;通过四唑盐比色法(MTT法)观察pCH510转染BIU-87对BCG细胞毒作用的影响。结果BIU-87细胞转染pCH510后阳性表达CH50多肽,对照组则不表达;CH50多肽阳性表达增强了BCG对BIU-87的细胞毒性作用(P<0.05)。结论转染pCH510后,BIU-87细胞表达CH50多肽,并因此促进了BCG对膀胱癌细胞的抑制作用。     

Intestinal trefoil factor (TFF 3) and pS2 (TFF 1), but not spasmolytic polypeptide (TFF 2) mRNAs are co-expressed in normal,hyperplastic, and neoplastic human breast epithelium

pS2–TFF 1 is expressed in breast cancers and has been investigated as a potential prognostic factor reflecting oestrogen dependence. The relationship to the expression of other trefoil peptides, human spasmolytic polypeptide (hSP–TFF 2) and intestinal trefoil factor (hITF/hPI.B–TFF 3) is documented here. Fifty-seven breast specimens were selected from surgical pathology archives and included five normal breasts (two lactating), seven benign proliferative lesions, 11 ductal carcinomas in situ (DCIS), three lobular carcinomas in situ (LCIS), 24 invasive ductal carcinomas (IDC), and seven invasive lobular carcinomas (ILC). The comparative distribution of trefoil mRNAs was assessed by in situ hybridization using ³⁵S-labelled riboprobes and immunohistochemical staining for pS2–TFF 1 and hSP–TFF 2. pS2–TFF 1 and hITF/hPI.B–TFF 3 mRNA were focally present at low signal intensity in normal and benign breast. Both pS2–TFF 1 and hITF/hPI.B–TFF 3 were expressed in all DCIS, LCIS and ILC, and 21/24 IDC. Overall, expression patterns of pS2–TFF 1 and hITF/hPI.B–TFF 3 coincided, but hITF/hPI.B–TFF 3 mRNA was usually found in a greater proportion of cells. Expression of hSP–TFF 2 peptide or mRNA was not detected in any of these cases. MCF 7 breast carcinoma cells also expressed hITF/hPI.B–TFF 3 and pS2–TFF 1 mRNAs but not hSP–TFF 2. hITF/hPI.B–TFF 3 co-expression with pS2–TFF 1 may act as a prognostic factor, but also raises questions about the regulatory pathway for pS2–TFF 1 hITF/hPI.B–TFF 3. Trefoil factors have effects on cell motility and spreading in vitro, and co-expression of hITF/hPI.B–TFF 3 with pS2–TFF 1 could be functionally significant if they form a heterodimer or compete for receptor binding. Absence of hSP–TFF 2 expression may be of equal relevance to tumour cell biology. © 1997 by John Wiley & Sons, Ltd.     

Heterogeneity in hereditary pancreatitis

Hereditary pancreatitis (HP) is the most common form of chronic relapsing pancreatitis in childhood, and may account for ∼25% of adult cases with chronic idiopathic pancreatitis. Recently, an arginine-histidine (R117H) mutation within the cationic trypsinogen gene was found in 5/5 families studied with HP. In this study we report on the results of linkage and direct mutational analysis for the common R117H mutation examined in 8 nonrelated families with hereditary pancreatitis. Two-point linkage analysis with the 7q35 marker D7S676, done initially in 4 families, yielded lod scores that were positive in 2, negative in one, and weakly positive in one. Direct mutational analysis of exon 3 of the cationic trypsinogen gene in 6 families showed that all symptomatic individuals tested were heterozygous for the R117H mutation. Also, several asymptomatic but at-risk relatives were found to be heterozygous for this mutation. Affected individuals in the remaining 2 families did not have the mutation. Radiation hybrid mapping using the Genebridge 4 panel assigned the trypsinogen gene to chromosome region 7q35, 2.9 cR distal to ETS WI-9353 and 3.8 cR proximal the dinucleotide repeat marker D7S676. The negative linkage and absence of the trypsinogen mutation in 2/8 families suggest locus heterogeneity in HP. Analysis of the R117H mutation is useful in identifying presymptomatic “at-risk” relatives and in genetic counseling. Also, it can be useful in identifying children and adults with isolated chronic idiopathic pancreatitis. Am. J. Med. Genet. 77:47–53, 1998. © 1998 Wiley-Liss, Inc.