肺癌组织中凋亡相关基因bcl-2、fasmRNA表达研究

为探讨 bcl-2、fas凋亡相关基因在肺癌组织中表达及与肿瘤临床病理学的关系 ,采用逆转录聚合酶链式反应检测 43例肺癌中 bcl-2及 fas m RNA的表达。结果显示 ,肺癌组织中 bcl-2 m RNA的阳性率为 48.8% ,癌旁组织中 bcl-2 m RNA的阳性表达率为85 .7% ,显著性高于正常组织和癌组织 (P<0 .0 5 ) ;在鳞癌和腺癌中 ,bcl-2 m RNA的阳性表达与肺癌的分化程度呈正相关 ,与肺癌临床分期呈负相关。fas m RNA在鳞癌、腺癌、小细胞癌及大细胞癌中的阳性率分别为 93.3%、88.9%、6 2 .5 %和 0 % ,肺癌组织中 fas m RNA的阳性率极为显著高于正常肺组织及癌旁组织 (P<0 .0 1) ;鳞癌和腺癌中 ,fas m RNA的阳性率显著性高于小细胞癌 (P<0 .0 5 ) ,且与分化程度呈负相关。fas m RNA阳性率有随临床分期增加而上升的趋势 ,但无显著性差异 (P>0 .0 5 )。     

增殖细胞核抗原表达与肺癌的关系

为探讨增殖细胞核抗原(PCNA)表达与肺癌分化、TNM分期和预后的关系,用鼠抗PCNA单克隆抗体,以SP免疫组织化学方法检测106例肺癌及癌旁肺组织中PCNA表达指数。在肺癌组织中PCNA指数,鳞癌为456%、腺癌为394%、小细胞癌为474%,均显著高于癌旁肺组织的29%(P<0001)。在肺鳞癌和腺癌中,癌组织分化越差,PCNA指数越高(P<005)。PCNA指数与肺鳞癌淋巴结转移和TNM分期相关不密切,而与肺腺癌淋巴结转移和TNM分期均密切相关(P<005)。无论是鳞癌还是腺癌,术后存活3年以上者PCNA指数明显低于3年以下者(P<005)。结果提示PCNA的表达指数可反映肺癌组织的增殖能力,对评估肺癌的分化和预后有重要参考作用。     

肺癌血管内皮生长因子表达的研究

为探讨血管内皮生长因子(VDGF)与肺癌血管形成、转移和增殖活性等因素的关系，采用SP免疫组织化学方法观察VEGF在94例肺癌组织的表达．结果表明．VEGF在肺癌组织表达的阳性率为70.2%，明显高于癌旁正常组织(P＜0．01)，VEGF表达阳性组其激血管计数值和只PCNA标记指数均高于阴性组(P&;lt;0.05)。腺癌VEGF的表达与转移、分期和分级等因素均无关(P＞0.05)；在鳞癌有淋巴结转移组VEGF的阳性率高于无转移组，Ⅲ～Ⅳ期癌高于Ⅰ～Ⅱ期癌(P＜0.05)，上述结果提示，VEGF表达与肺癌的发生有关，并能促进肺癌组织中的血管形成．使癌细胞的增殖活性升高；VEGF表达与肺鳞癌淋巴结转移和进展有关．可能是预后不良的标志。     

非小细胞肺癌环氧化酶Ⅱ表达及意义

目的 探讨环氧化酶Ⅱ(COX-2)在非小细胞肺癌(NSCLC)组织中的表达及其与NSCLC发生、发展关系.方法 采用免疫组织化学法检测45例NSCLC组织和癌旁正常组织标本COX-2的表达水平.结果 COX-2在NSCLC组织的表达率为64.4%(26/45),明显高于癌旁正常组织的31.1%(13/45),两者比较差异有统计学意义(P＜0.05).COX-2表达在不同性别、年龄及组织分化间差异无统计学意义(P＞0.05);COX-2表达在腺癌明显高于鳞癌(P＜0.05),直径＞3 cm肿瘤组织明显高于直径≤3 cm肿瘤组织(P＜0.05),有淋巴结转移者明显高于无淋巴结转移者(P＜0.05),TNM分期Ⅲ+Ⅳ期者明显高于Ⅰ+Ⅱ期者(P＜0.05).结论 COX-2参与了肺癌发生、发展过程.     

非小细胞肺癌组织中Smad7表达谱的改变

目的研究在非小细胞肺癌组织中转化生长因子(TGF)-β1信号抑制子Smad7表达水平的变化,探讨Smad7在肺癌发生中的作用。方法收集46例临床肺鳞癌、腺癌标本,用免疫组织化学方法检测肺癌及癌旁组织中Smad7的表达丰度,比较表达差异,并取肿瘤组织检测为阳性的12例鳞癌病例标本,分别提取其癌组织及癌旁组织总蛋白,用Westernblot进行验证。结果肺癌组织中Smad7表达总阳性率为44%(20/46),显著高于癌旁组织17%(8/46()χ2=7.91,P<0.01);其中,鳞癌和腺癌癌组织中Smad7的阳性率分别为60%(12/20)和31%(8/26),癌旁组织中Smad7的阳性率分别为25%(5/20)和12%(3/26)。肺鳞癌组织中Smad7的表达高于癌旁组织(χ2=5.01,P<0.05)。Westernblot验证结果表明,免疫组织化学检测阳性的12例标本癌组织Smad7Westermblot检测均为阳性,其中有7例癌组织Smad7表达高于癌旁组织,进一步验证了Smad7蛋白在肺鳞癌中表达增高。结论Smad7的过表达同肺鳞癌的发生相关。     

clusterin、bcl-2在非小细胞肺癌中的表达及其相关性

目的:研究clusterin、bcl-2在非小细胞肺癌(NSCLC)组织中的表达及其相关性,探讨其与非小细胞肺癌发生、发展的关系.方法:免疫组化pv9000通用型二步法检测53例NSCLC组织、20例癌旁组织中簇集蛋白(Clusterin)、bcl-2的阳性表达情况.结果:非小细胞肺癌组clusterin、bcl-2阳性表达率高于癌旁组织(P＜0.05);clusterin、bcl-2表达与肺癌TNM分期、分化程度及淋巴结转移均有关(P＜0.05),但与组织学类型、性别、年龄无关(P＞0.05);在非小细胞肺癌组织中,clusterin、bcl-2表达成正相关(r=0.396,P＜0.05).结论:clusterin和bcl-2的高表达可能与非小细胞肺癌的发生、发展和浸润有关,二者在肺癌的发生发展中可能存在协同作用.     

COX-2和Bax在非小细胞肺癌中的表达及意义

目的探讨非小细胞肺癌（NSCLC）中环氧化酶-2（COX-2）和Bax的表达及其临床意义。方法采用免疫组织化学方法检测57例NSCLC癌组织（腺癌/鳞癌）、相应配对的15例正常组织及57例淋巴结COX-2蛋白和Bax蛋白的表达水平,计算其阳性百分比进行相关性分析。结果NSCLC组织中COX-2蛋白表达高于癌旁正常组织（P〈0.05）,腺癌高于鳞癌（P〈0.05）,与肿瘤组织的病理分级相关,且与淋巴结转移呈正相关（P〈0.05）,但与患者的年龄、性别无明显相关（P〉0.05）;Bax蛋白表达低于癌旁正常组织（P〈0.05）,腺癌高于鳞癌（P〈0.05）,且与淋巴结转移呈负相关（P〈0.05）,与年龄、性别、病理分级间差异无统计学意义（P〉0.05）。结论COX-2在NSCLC尤其是肺腺癌中表达上调,Bax蛋白在NSCLC组织中表达过低,COX-2可能通过降低Bax表达,抑制细胞凋亡参与NSCLC的发生、发展。     

RASSF10 mRNA在肺癌组织中表达的研究

目的探讨RASSF10基因在肺癌组织中的表达情况。方法采用反转录聚合酶链反应(RTPCR)技术检测RASSF10mRNA在37例肺癌组织及15例癌旁正常组织中的表达情况,37例肺癌组织中,鳞状细胞癌18例、腺癌12例、小细胞肺癌7例。结果 RASSF10mRNA在癌旁正常组织中的表达阳性率为93%(14/15),明显高于其在肺癌组织中的19%(7/37)(P<0.05);且在肺癌组织中,RASSF10 mRNA在鳞状细胞癌、腺癌、小细胞癌中的阳性表达率分别为22%(4/18)、25%(3/12)、0(0/7),差异无统计学意义(P均>0.05)。结论 RASSF10基因的异常改变可能参与了肺癌的发生发展过程;RASSF10可作为肺癌诊断的新的肿瘤标志,并可能成为新的治疗靶点。     

肺癌MMP-9和VEGF的表达及意义

目的 探讨肺癌组织中基质金属蛋白酶-9(MMP-9)和血管内皮生长因子(VEGF)的表达及其意义.方法 采用免疫组化S-P法检测20例癌旁正常肺组织、84例肺癌组织中MMP-9和VEGF的表达.结果 肺癌组织MMP-9和VEGF表达显著高于癌旁正常肺组织(P<0.01).小细胞肺癌和腺癌MMP-9表达高于鳞癌(P<0.01).肺癌MMP-9表达有淋巴结转移者高于无淋巴结转移者(19<0.01),Ⅲ期和Ⅳ期高于Ⅰ期和Ⅱ期(P<0.05).肺癌VEGF表达有淋巴结转移者高于无淋巴结转移者(P<0.05),Ⅲ期和Ⅳ期高于Ⅰ期和Ⅱ期(P<0.05).肺癌MMP-9和VEGF的表达呈正相关(P<0.05).结论 MMP-9和VEGF表达与肺癌侵袭转移密切相关.     

EGFR在子宫颈癌及淋巴结转移灶中的表达

目的 探讨 EGFR 在宫颈癌组织及淋巴结转移灶中的表达与肿瘤病理及淋巴转移发生的关系。方法 用免疫组化法检测 88例宫颈浸润癌,35例宫颈上皮内瘤样病变( ) 例正常宫颈组织及 14例淋巴结转移灶中的 EG FR 。 CIN ,20结果 正常宫颈鳞状上皮 EGFR 阳性率为 15%,CIN Ⅰ、CIN Ⅱ和 CIN Ⅲ EGFR 阳性率分别为 55.6%、50.0%和 73.3%,鳞癌为 42.6%,腺癌为 20%。其中宫颈鳞癌及 CIN 各级较正常宫颈 EGFR 阳性率显著增高(P <0.05)鳞癌较腺癌显著升高 ,(P <0.05) ;鳞癌与 CIN 各级间 EGFR 阳性率均无显著差异 (P>0.05) ;宫颈鳞癌 EGFR 表达与癌组织分级有关,其高分化( ) GI 、中分化( Ⅱ)和低分化( Ⅲ)宫颈癌 EGFR 阳性率分别为 64.3%、59.1%和 18.8%,其中 GI和 G Ⅱ较 G Ⅲ有显著 G G差异性(P <0.05) 。有淋巴转移宫颈癌原发灶中 EGFR 的阳性率为 71.4%,较无淋巴结转移肿瘤原发灶的 35.2%有显著差异性 (P <0.05),比较同一病例原发灶及转移灶间 EGFR 的阳性率无显著性差异(P>0.05) ,经等级相关检验显示两者EGFR 表达有显著相关性(P <0.05。结论 EGFR 可能与宫颈鳞癌发生有关,尤其在癌变前期和癌变初期起重要作用     

Safety assessment of poloxamers 101, 105, 108, 122, 123, 124, 181, 182, 183, 184, 185, 188, 212, 215, 217, 231, 234, 235, 237, 238, 282, 284, 288, 331, 333, 334, 335, 338, 401, 402, 403, and 407, poloxamer 105 benzoate, and poloxamer 182 dibenzoate as used in cosmetics

Poloxamers are polyoxyethlyene, polyoxypropylene block polymers. The impurities of commercial grade Poloxamer 188, as an example, include low-molecular-weight substances (aldehydes and both formic and acetic acids), as well as 1,4-dioxane and residual ethylene oxide and propylene oxide. Most Poloxamers function in cosmetics as surfactants, emulsifying agents, cleansing agents, and/or solubilizing agents, and are used in 141 cosmetic products at concentrations from 0.005% to 20%. Poloxamers injected intravenously in animals are rapidly excreted in the urine, with some accumulation in lung, liver, brain, and kidney tissue. In humans, the plasma concentration of Poloxamer 188 (given intravenously) reached a maximum at 1 h, then reached a steady state. Poloxamers generally were ineffective in wound healing, but were effective in reducing postsurgical adhesions in several test systems. Poloxamers can cause hypercholesterolemia and hypertriglyceridemia in animals, but overall, they are relatively nontoxic to animals, with LD(50) values reported from 5 to 34.6 g/kg. Short-term intravenous doses up to 4 g/kg of Poloxamer 108 produced no change in body weights, but did result in diffuse hepatocellular vacuolization, renal tubular dilation in kidneys, and dose-dependent vacuolization of epithelial cells in the proximal convoluted tubules. A short-term inhalation toxicity study of Poloxamer 101 at 97 mg/m(3) identified slight alveolitis after 2 weeks of exposure, which subsided in the 2-week postexposure observation period. A short-term dermal toxicity study of Poloxamer 184 in rabbits at doses up to 1000 mg/kg produced slight erythema and slight intradermal inflammatory response on histological examination, but no dose-dependent body weight, hematology, blood chemistry, or organ weight changes. A 6-month feeding study in rats and dogs of Poloxamer 188 at exposures up to 5% in the diet produced no adverse effects. Likewise, Poloxamer 331 (tested up to 0.5 g/kg day(-1)), Poloxamer 235 (tested up to 1.0 g/kg day(-1)), and Poloxamer 338 (at 0.2 or 1.0 g/kg day(-1)) produced no adverse effects in dogs. Poloxamer 338 (at 5.0 g/kg day(-1)) produced slight transient diarrhea in dogs. Poloxamer 188 at levels up to 7.5% in diet given to rats in a 2-year feeding study produced diarrhea at 5% and 7.5% levels, a small decrease in growth at the 7.5% level, but no change in survival. Doses up to 0.5 mg/kg day(-1) for 2 years using rats produced yellow discoloration of the serum, high serum alkaline phosphatase activity, and elevated serum glutamicpyruvic transaminase and glutamic-oxalacetic transaminase activities. Poloxamers are minimal ocular irritants, but are not dermal irritants or sensitizers in animals. Data on reproductive and developmental toxicity of Poloxamers were not found. An Ames test did not identify any mutagenic activity of Poloxamer 407, with or without metabolic activation. Several studies have suggested anticarcinogenic effects of Poloxamers. Poloxamers appear to increase the sensitivity to anticancer drugs of multidrug-resistant cancer cells. In clinical testing, Poloxamer 188 increased the hydration of feces when used in combination with a bulk laxative treatment. Compared to controls, one study of angioplasty patients receiving Poloxamer 188 found a reduced myocardial infarct size and a reduced incidence of reinfarction, with no evidence of toxicity, but two other studies found no effect. Poloxamer 188 given to patients suffering from sickle cell disease had decreased pain and decreased hospitilization, compared to controls. Clinical tests of dermal irritation and sensitization were uniformly negative. The Cosmetic Ingredient Review (CIR) Expert Panel stressed that the cosmetic industry should continue to use the necessary purification procedures to keep the levels below established limits for ethylene oxide, propylene oxide, and 1,4-dioxane. The Panel did note the absence of reproductive and developmental toxicity data, but, based on molecular weight and solubility, there should be little skin penetration and any penetration of the skin should be slow. Also, the available data demonstrate that Poloxamers that are introduced into the body via routes other than dermal exposure have a rapid clearance from the body, suggesting that there would be no risk of reproductive and/or developmental toxicity. Overall, the available data do not suggest any concern about carcinogenesis. Although there are gaps in knowledge about product use, the overall information available on the types of products in which these ingredients are used, and at what concentration, indicates a pattern of use. Based on these safety test data and the information that the manufacturing process can be controlled to limit unwanted impurities, the Panel concluded that these Poloxamers are safe as used.     

HPLC法测定抗妇炎胶囊中木兰花碱、黄柏碱、药根碱、巴马汀、小檗碱、槐果碱、苦参碱、氧化槐果碱、槐定碱和氧化苦参碱

目的 采用高效液相色谱（HPLC）法同时测定抗妇炎胶囊中木兰花碱、黄柏碱、药根碱、巴马汀、小檗碱、槐果碱、苦参碱、氧化槐果碱、槐定碱和氧化苦参碱10种活性成分。方法 采用InerSustain AQ-C₁₈色谱柱（250 mm×4.6 mm,5 μm）,流动相A：乙腈–无水乙醇（80∶20）,流动相B：0.1%磷酸溶液,梯度洗脱,检测波长220 nm,体积流量1.0 mL/min,柱温30℃,进样量10 μL。结果 木兰花碱、黄柏碱、药根碱、巴马汀、小檗碱、槐果碱、苦参碱、氧化槐果碱、槐定碱和氧化苦参碱分别在2.69～134.50、1.95～97.50、0.63～31.50、0.86～43.00、11.95～597.50、0.59～29.50、6.08～304.00、4.85～242.50、1.66～83.00、19.79～989.50 μg/mL线性关系良好（r≥0.999 3）;平均回收率分别为99.11%、98.23%、96.95%、97.78%、100.02%、97.21%、99.66%、99.52%、98.81%、100.08%,RSD值分别为1.04%、1.23%、1.37%、1.65%、0.70%、1.28%、0.65%、0.81%、1.11%、0.63%。结论 建立的HPLC法可用于抗妇炎胶囊中10种活性成分的测定,作为抗妇炎胶囊质量控制方法。     

Arformoterol: (R,R)-eformoterol, (R,R)-formoterol, arformoterol tartrate, eformoterol-sepracor, formoterol-sepracor, R,R-eformoterol, R,R-formoterol

Sepracor in the US is developing arformoterol [R,R-formoterol], a single isomer form of the beta(2)-adrenoceptor agonist formoterol [eformoterol]. This isomer contains two chiral centres and is being developed as an inhaled preparation for the treatment of respiratory disorders. Sepracor believes that arformoterol has the potential to be a once-daily therapy with a rapid onset of action and a duration of effect exceeding 12 hours. In 1995, Sepracor acquired New England Pharmaceuticals, a manufacturer of metered-dose and dry powder inhalers, for the purpose of preparing formulations of levosalbutamol and arformoterol. Phase II dose-ranging clinical studies of arformoterol as a longer-acting, complementary bronchodilator were completed successfully in the fourth quarter of 2000. Phase III trials of arformoterol began in September 2001. The indications for the drug appeared to be asthma and chronic obstructive pulmonary disease (COPD). However, an update of the pharmaceutical product information on the Sepracor website in September 2003 listed COPD maintenance therapy as the only indication for arformoterol. In October 2002, Sepracor stated that two pivotal phase III studies were ongoing in 1600 patients. Sepracor estimates that its NDA submission for arformoterol, which is projected for the first half of 2004, will include approximately 3000 adult subjects. Sepracor stated in July 2003 that it had completed more than 100 preclinical studies and initiated or completed 15 clinical studies for arformoterol inhalation solution for the treatment of bronchospasm in patients with COPD. In addition, Sepracor stated that the two pivotal phase III studies in 1600 patients were still progressing. In 1995, European patents were granted to Sepracor for the use of arformoterol in the treatment of asthma, and the US patent application was pending.     

欧洲刺柏（Juniper）

活性成分与药理作用欧洲刺柏药用部位是其浆果，具有促水排泄、防腐、抗胃肠胀气和抗风湿作用，还可改善胃功能。用作促水排泄药可增加尿量（水丢失），但不增加钠排泄。成分萜品烯-4-醇可增加肾小球滤过率，但刺激肾。欧洲刺柏浆果对单纯疱疹病毒体外显示抗病毒活性，并具抗真菌活性。动物实验显示，欧洲刺柏浆果提取物具有堕胎、抗生育、抗炎、抗胚胎植入、降血压、升血压和降血糖作用。欧洲刺柏浆果油具有兴奋子宫的活性，以及利尿、胃肠道抗菌和刺激作用，该油对平滑肌有阻止解痉作用。     

Oral Presentations (LDD, LPES, LNM, LPAT, LNT, LPPT, LPM)

Probiotic microorganisms have been shown to provide specific health benefits when consumed as food supplements or as food components. The main problem of such products is the poor survival of the probiotic bacteria in the low pH of gastric fluid. However the use of synthetic excipients for enteric coating to prevent the exposure of microorganisms to gastric fluid is limited in food supplementary industry. Therefore the aim of this study was to develop an enteric coating formulation containing shellac as a natural polymer. Shellac possesses good resistance to gastric juice; the major disadvantage of this polymer is its low solubility in the intestinal fluid [1, 2]. Thus films containing different ratios of shellac and water-soluble polymers (sodium alginate, hydroxypropyl methylcellulose (HPMC) and polyvinylpyrrolidon (PVP)) or plasticizers (glycerol and glyceryl triacetate (GTA)) were prepared in order to analyse the films’ melting temperatures (T_m), the changes in enthalpy (ΔH), their capability of taking up water, and their solubility in different media. The release characteristics of the films were studied by loading pellets with Enterococcus faecium M74 and coating them with formulations containing different amounts of shellac and polymer or plasticized shellac. Using dissolution tests, performed according to USP XXXI paddle method, the resistance of the coatings to simulated gastric fluid (SGF, pH 1.2) and the release of cells in simulated intestinal fluid (SIF, pH 6.8) was investigated.The trials showed that an increasing amount of plasticizer results in a decrease of T_m and ΔH of the films whereat glycerol had a superior plasticization effect to GTA. The compatibility of films made of water-soluble polymers and shellac was also concentration dependent. HPMC and PVP showed superior compatibility with shellac compared to sodium alginate, since films containing shellac and more than 10% [w/w] sodium alginate tended to separate into two phases. In the end five formulations containing shellac and either 5% [w/w] glycerol, 10% [w/w] PVP, 20% [w/w] PVP, 10% [w/w] HPMC, or 5% [w/w] sodium alginate emerged as feasible for enteric coating purposes.