蛋白激酶Cα和半胱氨酸天冬氨酸蛋白酶3在大肠癌中的表达及其与细胞凋亡的关系

目的探讨蛋白激酶Ca（PKC—α）与半胱氨酸天冬氨酸蛋白酶3（Caspase-3）在大肠癌中的表达及其与细胞凋亡的关系。方法采用免疫组化SP法检测83例大肠组织（正常组织15例,腺瘤组织21例,癌组织47例）中PKC-α和Caspase-3的表达水平,同时采用DNA原位末端标记法标记大肠癌组织中的凋亡细胞。结果PKC-α在大肠癌组织中阳性表达[31／47例（66．0％）]与正常组织[3／15例（20．0％）]及腺瘤[10／21例（47．6％）]相比,差异有统计学意义（P〈0．05）。Caspase-3在正常组织中的阳性表达[12／15例（80．0％）],与腺瘤[13／21例（61．9％）]及大肠癌组织[15／47例（31．9％）]相比,差异有统计学意义（P〈0．05）,Caspase-3的阳性表达仅与大肠癌组织分化程度关系密切（P〈0．05）。PKC-仅阳性表达的大肠癌组织中细胞凋亡指数（1．62±0．57）明显低于阴性组（3．28±0．75）（P〈0．05）,Caspase-3阳性组细胞凋亡指数（2．40±0．55）明显高于阴性组（1．25±0．59）（P〈0．05）。PKC—α与Caspase-3的表达呈负相关（r=-0．498,P〈0．05）。结论PKC—α和Caspase-3与大肠癌的细胞凋亡密切相关,参与细胞凋亡的调节机制,在大肠癌恶变的过程中发挥重要作用。     

基质金属蛋白酶-9任甲状腺良恶性病变组织中表达及临床意义

目的：探讨甲状腺良恶性病变组织中基质金属蛋白酶-9（MMP-9）的表达及其临床意义。方法：应用微波-LSAB免疫组化法检测50例甲状腺癌、45例甲状腺腺瘤和20例癌旁甲状腺组织中MMP-9的表达。结果：甲状腺癌中MMP-9表达阳性率（64．0％）显著高于甲状腺腺瘤（37．8％）和癌旁甲状腺组织（25．0％，P〈0．05）。MMP-9表达阳性率与甲状腺癌组织类型无关；有淋巴结转移者显著高于无淋巴结转移者（P〈0．05）；病理分期Ⅲ～Ⅳ期病例显著高于Ⅰ～Ⅱ期病例（P〈0．05）；MMP-9阳性的甲状腺癌复发及病死率显著高于阴性者（P〈0．05）。结论：MMP-9表达对甲状腺癌恶性程度判断、生物学行为预测和预后评估有一定价值。     

MTA1、MMP-9和TIMP-1蛋白与神经母细胞瘤侵袭转移相关性研究

目的探讨MTA1、MMP-9和TIMP-1蛋白与小儿神经母细胞瘤生物学行为之间的关系。方法运用免疫组化S—P法检测56例神经母细胞瘤组织中MTA1、MMP-9、TIMP—1蛋白的表达。结果56例神经母细胞瘤组织中37例MTA1阳性表达,阳性率为66．1％,MTA1高表达与神经母细胞瘤临床分期和转移关系密切（P〈0．05）;39例MMP-9蛋白阳性表达,阳性率为69．6％,MMP-9高表达与神经母细胞瘤临床分期和转移关系密切（P〈0．05）;27例TIMP—1蛋白阳性表达,阳性率为48．2％,TIMP-1低表达与神经母细胞瘤临床分期和转移关系密切（P〈0．05）;神经母细胞瘤组织中MMP-9,TIMP-1蛋白表达呈负相关（P〈0．05）。结论MTA1、MMP-9和TIMP-1蛋白表达与神经母细胞瘤恶性程度、转移和预后关系密切,可作为判断预后的指标,并有望为肿瘤靶向治疗提供理论依据。     

神经母细胞瘤血管内皮细胞生长因子、Ki-67抗原表达及微血管密度研究

目的 探讨血管内皮细胞生长因子（VEGF）、瘤内微血管密度（MVD）与神经母细胞瘤（NB）细胞增殖及预后的关系。方法 采用S-P免疫组化法，检测VEGF、CD34和Ki-67抗原在21例NB中的表达及MVD计数。结果 VEGF及Ki-67表达与NB病理分型及核分裂指数有关（P〈0．05或P〈0．01），而与NB分化与否、肿瘤大小及性别无关。MVD值在临床各病理参数间差异无显著意义。VEGF表达与MVD值、VEGF与Ki-67表达呈明显正相关（P〈0．01）。结论 VEGF、Ki-67表达与NB细胞增殖及预后密切相关。     

胰腺癌中MMP- 2、VEGF-D 的表达及意义

目的探讨胰腺癌组织中MMP－2、VEGF－D的表达和相关性及其与，陆床病理学特征、预后的关系。方法免疫组化（SABC法）分别检测28例手术切除的胰腺癌组织和8例正常胰腺组织中MMP-2、VEGF-D的表达，分析二者的表达与临床病理特征、预后的关系。结果28例胰腺癌组织中MMP－2高表达者有18例（64．3％），其中强阳性5例（17．9％），阳性13例（46．4％），弱阳性6例（21．4％），阴性4例（14．3％）；vEGF-D高表达者16例（57．1％），其中强阳性4例（14．3％），阳性12例（42．9％），弱阳性7例（25．O％），阴性5例（17．8％），8例对照胰腺组织中两者表达均阴性。淋巴结转移组MMP－2表达明显强于无淋巴结转移组沪〈0．05），T，组MMP－2表达明显强于T1-2组（P〈0．05），MMP-2表达与病人生存率无明显相关性（P〉0．05），淋巴结转移组VEGF-D表达明显强于无淋巴结转移组（P〈0．05），VEGF-D高表达组与低表达组间生存率无显著性差异（P〉0．05）。MMP－2高表达与VEGF-D高表达正相关（r=0．553，P=0．003），MMP－2、VEGF－D均高表达组淋巴结转移率明显高于MMP-2、VEGF-D均低表达组（P〈0．05）。结论（1）MMP－2、VEGF－D在胰腺癌中均表达增强；（2）MMP-2在胰腺癌中的高表达与胰腺癌的局部浸润、淋巴结转移相关，与预后无关；（3）VEGF-D在胰腺癌中的高表达与胰腺癌的淋巴结转移相关，与预后无关（4）胰腺癌组织中VEGF-D与MMP-2表达呈正相关，两者在胰腺癌淋巴转移中可能起着重要协同作用。     

乳腺癌组织中脆性组氨酸三联体基因的表达及意义

目的探讨乳腺癌组织中脆性组氨酸三联体（FHIT）基因的表达及其临床意义。方法应用免疫组织化学酶联免疫sP法,检测FHIT在41例乳腺癌组织及11例癌旁组织中的表达。结果乳腺癌组FHIT蛋白阳性表达率46．3％（19／41）显著低于正常乳腺组90．9％（10／11）（P〈0．05）。中／高分化组FHIT蛋白阳性表达率显著高于低分化组（53．8％vs33．3％）（P〈0．05）;淋巴结转移组FHIT蛋白阳性率低于,无淋巴结转移组（42．9％vs53．8％）（P〈0．05）;Ⅰ＋Ⅱ期组与Ⅲ＋Ⅳ期组之间FHIT蛋白阳性表达率（56．3％ vs 40．0％）差异有统计学意义（P〈0．05）。结论FHIT基因在乳腺癌的发生、发展中起重要作用,对其蛋白的检测可作为判定乳腺癌发生及转移能力的一项客观指标。     

表皮生长因子受体和基质金属蛋白酶-9在胃癌组织中的表达及意义

目的探讨表皮生长因子受体（EGFR）和基质金属蛋白酶-9（MMP-9）在胃癌组织中的表达及临床意义。方法采用免疫组化SP法检测80例胃癌组织和30例正常胃组织中EGFR和MMP-9蛋白的表达情况。结果80例胃癌组织中44例EGFR表达阳性（55．O％）,明显高于正常胃组织中的6．7％（2／30）（X^2=20．95,P〈0．05）;在淋巴结转移阳性患者胃癌组织中的EGFR表达阳性率为63．6％（35／55）,高于淋巴结转移阴性患者胃癌组织的36．0％（9／25）（x。=5．30,P〈0．05）。MMP-9在80例胃癌组织中阳性表达率70．0％（56／80）,明显高于正常胃组织中的20．o％（6／30）（X^2=22．18,P〈0．05）;在胃癌不同TNM分期中,（Ⅲ期＋Ⅳ期）组中EGFR和MMP-9的表达均高于其在（Ⅰ期＋Ⅱ期）组中的表达（X^=4．26、5．78,均P〈0．05）。结论EGFR和MMP-9在胃癌组织中高表达,与胃癌TNM分期及淋巴结转移相关,是评估胃癌生物学行为及预后的重要指标。     

甲泼尼龙琥珀酸钠对自身免疫性肺气肿大鼠 TNF-a、MMP-9、IL-8及VEGF水平的影响

目的探讨糖皮质激素对自身免疫性肺气肿大鼠的肺泡灌洗液（BALF）中的肿瘤坏死因子-a（TNF-a）、基质金属蛋白酶-9（MMP-9）、白介素-8（IL-8）、血管内皮细胞生长因子（VEGF）水平的影响。方法将24只健康雄性SD大鼠随机分为正常对照组、模型组和干预组三组。建立自身免疫性肺气肿模型，干预组同时腹腔内注入糖皮质激素进行干预。21d后处死所有大鼠，三组大鼠均以ELSA法检测BALF中的TNF-a、MMP-9、Ib8和VEGF含量；三组均取肺组织切片进行苏木精-伊红（HE）染色观察病理形态学改变，并定量测定平均内皮衬间隔（MLI）、平均肺泡数（MAN）。结果（1）与正常对照组比较，模型组MLI增高（P〈0．05），MAN降低（P〈0．05）；与模型组比较，干预组MLI降低（P〈0．05），MAN增高（P〈0．05）。（2）模型组BALF中的TNF-a、MMP-9浓度高于正常对照组（P〈0．05），VEGF浓度低于正常对照组（P〈0．05）；干预组TNF-a、MMP-9浓度低于模型N（P〈0．05），VEGF浓度高于模型组（P〈0．05）；模型组和对照组间IL-8浓度差异无统计学意义（P〉O．05）。结论（1）TNF_d、MMP-9值升高及VEGF浓度降低可能促进了大鼠自身免疫性肺气肿的形成，Ib8可能不参与自身免疫性肺气肿的形成。（2）糖皮质激素可能抑制了自身免疫性肺气肿大鼠TNF-a、MMP-9的表达及VEGF的减少。     

膀胱癌中陷阱受体3、Bcl-2与PCNA的联合诊断价值

目的探讨膀胱癌中陷阱受体3（DcR3）,Bcl-2和PCNA及其联合检测的诊断价值。方法应用免疫组织化学技术检测224例膀胱癌组织标本和56例正常膀胱组织标本中DcR3,Bcl-2和PCNA的表达,并应用受试者工作特征曲线（ROC）分析三项指标及联合检测对膀胱癌诊断的临床价值。结果膀胱癌组中DcR3,Bcl-2和PCNA的表达均明显高于正常膀胱组织（均P〈0．05）。DcR3的表达与膀胱癌的病理分级、是否浸润、淋巴结转移和复发相关（P〈0．05）;Bcl-2的表达与膀胱癌的淋巴结转移相关（P〈0．05）;PCNA的表达与膀胱癌的病理分级和淋巴结转移相关（P〈0．05）。DcR3表达水平和Bcl-2及PCNA呈正相关关系,Bcl-2和PCNA也同样呈正相关关系（P〈0．05）。DcR3,Bcl-2及PCNA联合检测的曲线下面积（AUC）高于单独指标检测值。联合检测DcR3＋PCNA,Bcl-2＋PCNA及DcR3＋Bcl-2＋PCNA的灵敏度均优于单独检测,其中最高者为三者联合组,灵敏度为0．830。联合检测DcR3＋PCNA,Bcl-2＋PCNA及DcR3＋Bcl-2＋PCNA的阳性预测值分别为100％,96．2％,100％,也均高于单独检测。结论DcR3,Bcl-2,PCNA三者联合检测可显著提高膀胱癌诊断的灵敏度和阳性预测值,优于单项检测效果。     

胃癌及癌前病变中PCNA和Caspase-3的表达及其与Hp感染的关系

目的探讨PCNA和Caspase-3在胃癌及癌前病变中的表达及其与Hp感染的关系。方法应用快速尿素酶试验与吉姆斯染色法将各种病变分为Hp感染阳性组和阴性组,应用免疫组化法对各种病变胃组织PCNA和Caspase-3的表达进行检测。结果①胃癌及癌前病变组PCNA表达及Hp感染率明显高于慢性浅表性胃炎组（P〈0．05）,而Caspase-3的表达显著低于慢性浅表性胃炎组（P〈0．05）;②Hp感染阳性组PCNA及Caspase-3表达明显高于Hp感染阴性组（P〈0．05）。结论Hp感染可能通过促进PCNA及Caspase-3基因蛋白的过度表达,从而使胃黏膜细胞增生与凋亡平衡失调诱发癌变。     

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.