玻璃化法冷冻和慢速冷冻对人类卵巢组织的影响

目的比较玻璃化冷冻和慢速冷冻对人类卵巢组织的影响。方法采集19例人类卵巢组织,随机分成3组：玻璃化冷冻组、慢速程序冷冻组、新鲜对照组。对新鲜的和冻融后的卵巢皮质中的间质细胞和卵泡进行光学显微镜观察。同时,通过原位末端转移酶标记技术（TUNEL）实验观察卵泡凋亡的情况。结果慢速程序冷冻组异常卵巢间质细胞比率高于玻璃化冷冻组,差异有统计学意义（P〈0.05）。各冷冻组的异常始基卵泡及初级卵泡比率均高于新鲜对照组,差异有统计学意义（P〈0.05）。冷冻组与对照组的卵泡凋亡率差别无统计学意义（P〉0.05）。结论玻璃化冷冻是人类卵巢组织适宜的冷冻保存方案。     

不同玻璃化冷冻方法对家兔卵巢组织形态学c—kit和Ki67表达的影响

目的探讨最佳的玻璃化冷冻方案。方法12只家兔随机分配到新鲜组（A）、麦管组（B）、微滴组（C）及SSV组（D）后行组织形态学分析及免疫组化检测。结果A组始基卵泡的形态正常率为91．97％,与之相比．冷冻组B—D组冻融后始基细胞形态正常率分别是69．88％、71．22％、81．06％,较对照组明显下降,有显著性差异（P〈0．05）;D组中始基卵泡形态正常率最高,与B、C组相比差异有显著性（P〈0．05）;B、C组比较,差异无显著性（P〉0．05）;各冷冻组合并后始基卵泡的形态正常率为74．03％,初级卵泡形态正常率为64．66％,比较有统计学差异（P〈0．05）;免疫组化显示：冷冻组复苏后卵巢组织中始基和次级卵泡中c—kitR和Ki67的阳性表达率分别是63．07％、67．11％、66．67％及72．77％、75．46％、75．14％,与对照组（68．23％及77．83％）相比,均无统计学意义（P〉0．05）。结论3种玻璃化冷冻方法对卵巢皮质中各级卵泡及其结构均造成一定程度的损害,使各级卵泡的形态正常率明显下降,卵巢间质细胞连接变得疏松;SSV法明显优于其他两种玻璃化冷冻方法,较适合卵巢组织中始基卵泡的保存;冻存卵巢组织对初级卵泡的影响大于始基卵泡。三种玻璃化冷冻方法对家兔卵巢组织c—kit和Ki67的表达均无明显影响;三种冷冻复苏方法均能较好的保存家兔卵巢组织中卵泡生长发育的启动、增殖活性。     

米非司酮对子宫肌瘤组织中bcl-2、P53和雌、孕激素受体的影响

目的研究米非司酮对子宫肿瘤细胞内凋亡调控基因bcl-2、P53和雌激素受体(ER)、孕激素受体(PR)表达的影响.方法应用免疫组化技术测定12例经米非司酮治疗的子宫肌瘤组织中bcl-2、P53和ER、PR的阳性表达率,并以12例未用药的子宫肌瘤患者作对照.结果服药组子宫肌瘤组织中bcl-2、PR、ER阳性表达率较对照组显著下降(P＜0.01),P53阳性表达率两组差异无显著性(P＞0.05).结论米非司酮可明显降低肌瘤组织中ER、PR,而且可以抑制肌瘤细胞内bcl-2表达,从而诱导子宫肌瘤平滑肌细胞凋亡.     

57例乳癌中c—erbB—2、p53、ki—67表达与临床病理关系分析

目的 研究乳腺癌组织中的c—erbB—2及突变型p53癌基因和ki—67抗原表达与乳癌临床特点及激素受体关系，揭示c—erbB—2、p53、ki—67表达对乳腺癌预后的评估意义。方法 应用S—P免疫组化方法检测57例乳腺癌组织中c—erbB—2、p53、ki—67表达情况与临床病理参数的关系，结果作统计学分析。结果 (1)在57例乳腺癌组织中，c—erbB—2、p53、ki—67的阳性表达率分别为63．2％(36／57)、42．1％(24／57)和57．9％(33／57)。(2)ki—67表达随淋巴结转移率及肿瘤分期增加而增加。(3)ER、PR阳性患者的c—erbB—2的阳性表达明显低于ER、PR阴性患者。结论 乳腺癌c—erbB—2、p53、ki—67的高表达与肿瘤浸润或转移密切相关，ER、PR与c—erbB—2、p53、ki—67这两种不同特性产物在乳腺癌组织中有一定的内在联系。因此，检测这些指标有助于判断乳腺癌的恶性度和生物学行为，确定术后治疗方案。     

子宫平滑肌瘤中ER、PR及Survivin、Ki-67表达的研究

目的：通过对子宫平滑肌瘤中ER、PR及Survivin、Ki-67表达情况的检测分析,探讨子宫平滑肌瘤的发生机制及临床意义。方法：采用免疫组化法（S-P法）检测60例（30例普通型、30例富于细胞型）平滑肌瘤及40例正常子宫平滑肌组织中ER、PR、Survivin、Ki-67的表达情况。结果：子宫平滑肌瘤ER阳性率为71.7%,PR阳性率为86.7%,正常平滑肌组织中ER、PR阳性率分别为20.0%、67.5%,两组比较差异有统计学意义（P〈0.05）;子宫平滑肌瘤Survivin阳性率86.7%、Ki-67阳性率为73.3%,正常平滑肌组织中Survivin、Ki-67阳性率均为5%,两组相比差异有统计学意义（P〈0.05）。结论：ER、PR、Survivin、Ki-67均是反映子宫肌瘤生物学行为的指标,与子宫肌瘤的发生发展密切相关,且雌、孕激素与ER、PR结合后刺激肌瘤生长可能与改变细胞增殖与凋亡的平衡有关。     

不同玻璃化冷冻方法对家兔卵巢组织形态学c-kit和Ki67表达的影响

目的探讨最佳的玻璃化冷冻方案。方法12只家兔随机分配到新鲜组(A)、麦管组(B)、微滴组(C)及SSV组(D)后行组织形态学分析及免疫组化检测。结果A组始基卵泡的形态正常率为91.97%,与之相比,冷冻组B～D组冻融后始基细胞形态正常率分别是69.88%、71.22%、81.06%,较对照组明显下降,有显著性差异(P<0.05);D组中始基卵泡形态正常率最高,与B、C组相比差异有显著性(P<0.05);B、C组比较,差异无显著性(P>0.05);各冷冻组合并后始基卵泡的形态正常率为74.03%,初级卵泡形态正常率为64.66%,比较有统计学差异(P<0.05);免疫组化显示:冷冻组复苏后卵巢组织中始基和次级卵泡中c-kitR和Ki67的阳性表达率分别是63.07%、67.11%、66.67%及72.77%、75.46%、75.14%,与对照组(68.23%及77.83%)相比,均无统计学意义(P>0.05)。结论3种玻璃化冷冻方法对卵巢皮质中各级卵泡及其结构均造成一定程度的损害,使各级卵泡的形态正常率明显下降,卵巢间质细胞连接变得疏松;SSV法明显优于其他两种玻璃化冷冻方法,较适合卵巢组织中始基卵泡的保存;冻...     

乳腺癌患者超声声像图特征与PR、ER、ki67表达的相关性研究

目的探讨乳腺癌患者超声声像图特征与孕激素受体(PR)、雌激素受体(ER)和Ki-67表达的相关性。方法选择78例乳腺癌患者,采用彩色多普勒超声检查乳腺癌患者肿物的直径、毛刺征、钙化、淋巴结转移情况。选取乳腺癌患者组织病理材料,采用免疫组化法测定标本中PR、ER、ki67表达水平。并对对患者不同声像图特征下的PR、ER、ki67表达水平及相关性进行研究。结果不同直径肿瘤ER阳性表达水平间差异有统计学意义(χ2=15.9250,P<0.05),Ki-67表达水平间差异有统计学意义(χ2=10.5978,P<0.05)。乳腺癌肿瘤直径大小与Ki-67表达呈正相关(r=0.3686,P<0.05)。有无毛刺征PR阳性表达水平间差异有统计学意义(χ2=8.5791,P<0.05),ER阳性表达水平间差异有统计学意义(χ2=10.2428,P<0.05),Ki-67表达水平间差异无统计学意义(χ2=0.044,P>0.05)。淋巴结是否转移PR阳性表达水平间差异有统计学意义(χ2=4.7815,P<0.05),ER阳性表达水平间差异有统计学意义(χ2=12.0394,P<0.05),Ki-67表达水平间差异无统计学意义(χ2=4.8851,P<0.05)。乳腺癌肿瘤淋巴结是否转移与PR、ER表达呈负相关(r值分别为-0.2476和-0.3929,P<0.05),但与Ki-67表达呈正相关(r=0.2503,P<0.05)。结论乳腺癌患者超声声像图特征与PR、ER、ki67有一定的相关性,临床工作中可以根据患者超声声像图的表现对患者的病情、转归、预后进行评估,在一定程度上指导临床治疗,也可以对乳腺癌患者进行粗略筛查。     

雌激素受体、孕激素受体与凋亡基因蛋白在子宫腺肌症中的表达

目的探讨雌激素受体（ER）、孕激素受体（PR）及凋亡基因蛋白（bcl-2）在子宫腺肌症中正位及异位内膜的表达及意义。方法采用免疫组化PV-9000二步法检测20例正常子宫内膜,58例子宫腺肌症正位及异位内膜ER、PR和bcl-2的表达情况。结果子宫腺肌症病例中,正位和异位内膜组织中ER、PR、Bcl-2都有阳性表达,异位腺体上皮阳性率高于间质细胞（P〈0．05）,异位内膜Bcl-2阳性表达率高于正位内膜（P〈0．05）;且增生期阳性表达率高于分泌期,差异有显著性（P〈0．05）。子宫腺肌症病例正位和异位内膜中ER、PR阳性表达率差异无显著性（P〉0．05）,在异位内膜中ER、PR、bcl-2的表达具有相关性（P〈0．01）。结论子宫腺肌症正位和异位子宫内膜中均有ER、PR和bcl-2的阳性表达,提示可能与子宫腺肌症的发生发展有关。     

乳腺导管内癌及其微浸润癌病理和ER、PR、p53和Ki-67表达差异分析

目的：探讨乳腺导管内癌及其微浸润癌病理表达情况。方法：通过对我院乳腺导管内癌及其微浸润癌患者25例临床资料进行分析,对乳腺导管内癌和微浸润癌ER、PR、p53和Ki-67表达情况进行比较观察。结果：微浸润癌ER、PR表达均低于乳腺导管内癌,乳腺导管内癌的Ki67明显低于微浸润癌表达,P〈0.05,差异有统计学意义,两种癌变的p53蛋白差异无统计学意义,P〉0.05。结论：乳腺导管内癌进展和ER、PR、p53和Ki-67异常表达有着密切的关系,尤其是Ki-67敏感性较高,可以作为预测肿瘤细胞增殖活性增强的预警指标。     

三种冷冻方法对胎儿卵巢组织凋亡相关蛋白表达的Bcl-2、Bax、Caspase影响

目的观察三种冷冻方法对胎儿卵巢组织卵泡凋亡蛋白的影响。方法分别采用程序冻存法(PFG)、快速冻存法(RFG)、玻璃化冻存法(VG)冻存胎儿卵巢组织,解冻后采用免疫组织化学法检测凋亡相关蛋白Bcl-2、Bax、Caspase-3的表达。结果各冷冻组卵泡Bcl-2蛋白阳性表达率下降,与新鲜组相比差异显著(P<0.05);Bax蛋白阳性表达率上升,PFG、VG组Bax蛋白阳性表达与新鲜组(FCG)差别不显著(P>0.05);RFG显著高于FCG、PFG、VG(P<0.001)。冷冻后,Caspase-3蛋白阳性表达率上升,PFG、RFG显著高于FCG(P<0.05),VG与FCG差别不显著(P>0.05)。结论玻璃化冻存法可有效冻存胎儿卵巢组织。     

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.