34βE12 CD10及SMMHC在乳腺良恶性病变鉴别诊断中的意义

目的探讨乳腺肌上皮细胞标记物平滑肌肌球蛋白重链(SMMHC)、34βE12、CD10在乳腺良、恶性上皮性病变鉴别诊断中的意义。方法用免疫组织化学方法检测肌上皮细胞标记物SMMHC、34βE12及CD10在60例乳腺良、恶性病变中的表达,分析其与乳腺良、恶性上皮病变的关系及意义。结果34βE12、CD10和SMMHC在乳腺良性病变中的表达率均为100%;在乳腺癌中的表达率分别为26%、30%、35%;三者阳性表达率及其强度在良性病变与癌组之间差异有统计学意义(P<0.05);并且在乳腺癌中,SMMHC与34βE12和CD10相比较差异有统计学意义(P<0.05)。结论SMMHC、34βE12及CD10可作为乳腺病变鉴别诊断中肌上皮细胞的标记物,但SMMHC在标记乳腺肌上皮细胞时特异性及敏感性均优于34βE12和CD10。     

CD133和Nestin在乳腺癌中的表达

目的探讨CD133和Nestin在不同类型乳腺癌中的表达及其相互关系。方法应用免疫组织化学SP法检测38例乳腺浸润性导管癌、20例乳腺浸润性小叶癌、10例乳腺良性病变组织中CD133和Nestin的表达情况。结果CD133和Nestin在乳腺癌和乳腺良性肿瘤中均表达于胞浆;CD133和Nestin在乳腺癌中的表达阳性率分别为89．7％和72．4％,均高于良性肿瘤的50．0％和0;乳腺浸润性导管癌中CD1333,和Nestin的阳性表达率显著高于小叶癌,且CD133与Nestin表达之间呈正相关;CD133、Nestin阳性细胞率均与患者年龄、部位无关。结论CD133和Nestin在乳腺癌病理组织中均呈高表达,而在乳腺良性病变中呈低表达。CD133和Nestin的表达对乳腺癌的诊断具有较显著的临床意义。     

乳腺癌和导管内增生性病变CD105的表达及其临床意义

目的 探讨CD105在乳腺癌和乳腺导管内增生性病变中的表达及其作为肿瘤新生血管标志物的临床价值.方法 检测70例乳腺癌、64例乳腺导管内增生性病变和18例正常乳腺组织中CD105及三种促血管生成因子VEGF、bFGF和PDGF的表达,并对其与临床病理因素进行分析.结果 CD105标记的微血管密度(MVD)各组间差异均有统计学意义,病变区周围MVD明显高于病变区内的MVD,差别具有统计学意义;组织学分化程度低、临床TNM分期高、淋巴结转移阳性以及VEGF、bFGF和PDGF表达阳性的患者中,CD105表达明显升高,差别均有统计学意义.结论 CD105主要在处于增殖状态的血管内皮细胞上表达,在标记肿瘤组织新生血管方面的特异性优于泛血管内皮细胞标志物,可作为乳腺癌预后的重要指标之一.     

乳腺癌易感基因1在散发性乳腺癌组织中的表达

目的 探讨乳腺癌易感基因（BRCA1）蛋白在散发性乳腺癌组织中的表达.方法 收集86例乳腺癌及癌旁组织、48例乳腺增生症手术标本,采用免疫组化S-P法检测BRCA1蛋白的表达情况.结果 BRCA1表达在乳腺癌、乳腺癌旁组织、乳腺增生症中的阳性率分别为11.6%（10/86）、16.3%（14/86）和100%（48/48）,三组间比较差异有统计学意义（P＜0.01）.结论 BRCA1蛋白异常表达在散发性乳腺癌发生发展过程中起着一定作用.     

乳腺浸润性导管癌微血管生成与转录因子Etsl表达之间的关系

目的:探讨乳腺导管上皮癌变过程中微血管的分布特点及转录因子Etsl和其下游蛋白MMPI对肿瘤血管生成的作用.方法:应用免疫组织化学SP法检测正常乳腺18例(NB组)、导管上皮普通性增生20例(UDH组)、非典型增生29例(ADH组)、导管原位癌15例(DCIS组)和浸润性导管癌70例(IDC组)中CD34标记的微血管密度(MVD)、转录因子Ets1和其下游蛋白MMP1的表达,并应用CMIAS计算机辅助病理图像分析系统对结果进行分析.结果:CD34标记的MVD随着导管上皮增生病变程度的加重依次增加,差别具有统计学意义(均P<0.01);转录因子Ets1和其下游蛋白MMP1在IDC组中的表达明显高于导管内增生性病变(均P<0.01),且两者的表达呈正相关(r=0.55,P<0.01);IDC组中CD<,34>标记的MVD与Ets1和MMP1均有相关性(r分别为0.57和0.60,均P<0.01).结论:Ets1可能是调控乳腺癌血管生成的关键点.     

乳腺浸润性导管癌微血管生成与转录因子Ets1表达之间的关系

目的：探讨乳腺导管上皮癌变过程中微血管的分布特点及转录因子Ets1和其下游蛋白MMP1对肿瘤血管生成的作用。方法：应用免疫组织化学SP法检测正常乳腺18例（NB组）、导管上皮普通性增生20例（UDH组）、非典型增生29例（ADH组）、导管原位癌15例（DCIS组）和浸润性导管癌70例（IDC组）中CD34标记的微血管密度（MVD）、转录因子Ets1和其下游蛋白MMP1的表达,并应用CMIAS计算机辅助病理图像分析系统对结果进行分析。结果：CD34标记的MVD随着导管上皮增生病变程度的加重依次增加,差别具有统计学意义（均P〈0．01）;转录因子Ets1和其下游蛋白MMP1在IDC组中的表达明显高于导管内增生性病变（均P〈0,01）,且两者的表达呈正相关（r=0．55,P〈0．01）;IDC组中CD。标记的MVD与Ets1和MMP1均有相关性（r分别为0．57和0．60,均P〈0．01）。结论：Ets1可能是调控乳腺癌血管生成的关键点。     

Bmi-1在乳腺浸润性导管癌中的表达及临床意义

目的探讨Bmi-1在乳腺浸润性导管癌中的表达及其与临床病理特征之间的关系。方法采用免疫组织化学法检测60例乳腺浸润性导管癌组织中Bmi-1及Her-2、ER、PR的表达情况,并检测了30例乳腺增生症中Bmi-1表达情况。结果 60例乳腺浸润性导管癌中,Bmi-1蛋白高表达29例(48.3%),30例乳腺增生症中Bmi-1蛋白高表达6例(20.0%),两组之间差异有显著性(P<0.05)。Bmi-1蛋白阳性表达与乳腺浸润性导管癌的淋巴结转移及TNM分期密切相关(P<0.05),而与患者的年龄、肿瘤大小、组织学分级、Her-2、ER、PR等无显著性相关(P>0.05)。结论 Bmi-1蛋白在乳腺浸润性导管癌中的高表达与肿瘤进展相关,提示Bmi-1可能成为预测乳腺癌转移的新分子标志物。     

VCAM-1表达与乳腺癌血管生成及临床病理的关系

目的　研究乳腺癌瘤体中血管细胞粘附分子 (VCAM 1)及外周血清中可溶性血管细胞粘附分子 (sVCAM 1)的表达对乳腺癌血管生长及转移的影响。方法　对 5 2例乳癌瘤体、瘤旁 (2cm)及正常乳腺组织标本 ,2 0例良性乳房纤维瘤标本进行了VCAM 1、CD34免疫组化染色 ,图像分析 ,了解VCAM 1表达、CD34的表达情况 ,并分析VCAM 1的表达与微血管密度 (MVD)的相关性 ,同时用酶联免疫吸附试验对 2 5例正常人 ,30例乳腺癌术前、术后一周外周血进行检测 ,分析瘤体及外周血清中VCAM 1的表达与乳腺癌病理参数的关系。结果　VCAM 1不仅在乳腺癌血管内皮细胞上高表达 ,在乳腺癌细胞膜上也有表达 ,而且VCAM 1的表达强度、表达率与乳腺癌的血管的生成和转移相关。乳腺癌组病人外周血清中sVCAM 1的浓度比正常对照组高 (P <0 0 0 1) ,并且术后乳腺癌病人血清中sVCAM 1的浓度下降 (P <0 0 1)。结论　VCAM 1在乳腺癌的血管生成、癌细胞转移中可能起到重要作用 ,sVCAM 1有望成为乳腺癌的早期诊断及判断复发的指标。     

乳腺癌中VEGF-D的表达与微血管密度、微淋巴管密度的关系

目的探讨乳腺癌中血管内皮生长因子D(VEGF-D)的表达及其与肿瘤血管生成、淋巴管生成之间的关系.方法应用免疫组化法检测50例乳腺癌、10例乳腺增生症患者组织中的V EG F-D的表达 ,并以CD 105、D2-40作标记测定乳腺癌组织中的微血管密度(MVD)、微淋巴管密度(MLD).结果 乳腺癌组VEGF-D的阳性率及MVD、MLD明显高于乳腺增生症组(P<0.05) ,VEGF-D及 MVD、MLD 与乳腺癌组织学分级有相关性(P<0.05) ,MVD和MLD还与淋巴结转移密切相关(P<0.05) ,且VEGF-D与MVD、MLD之间呈正相关(P<0.05).结论 VEGF-D在乳腺癌的发生发展中具有重要作用 ,可促进肿瘤的微血管生成和微淋巴管生成 ,MVD和MLD与肿瘤的分化程度、侵袭能力和转移密切相关 ,可作为预测预后和转移的指标.     

细胞周期调节因子在乳腺导管增生性疾病及乳腺癌中的表达及意义

目的研究共济失调毛细血管扩张症突变蛋白(Ataxia-Telangiectasa mutated protein,ATM protein)、检测点激酶2(Checkpointskinase1,Chk2)及P53在乳腺浸润性导管癌(invasive ductal carcinoma,IDC)、导管内癌(ductal carcinoma in situ,DCIS)、非典型导管增生(atypical ductal hyperplasia,ADH)、癌旁正常乳腺组织(normal brease tissues adjacent to cancer,NBTAC)中的表达情况,探讨其在乳腺癌发生发展中的作用。方法采用免疫组化S-P法检测63例乳腺浸润性导管癌、39例导管内癌、42例非典型导管增生、56例癌旁正常乳腺组织中ATM、Chk2及P53的表达情况。结果 ATM、Chk2在乳腺浸润性导管癌中的表达明显低于癌旁正常乳腺组织;P53在乳腺浸润性导管癌中的表达明显高于癌旁正常乳腺组织;ATM、Chk2与P53表达呈显著负相关。结论乳腺浸润性癌ATM、Chk2表达缺失及P53的表达增高可能是乳腺癌发生、发展的重要因素之一,联合检测对早期乳腺癌的诊断及判断乳腺癌预后有指导意义。     

Interaction of estrone and estradiol with DNA and protein of liver and kidney in rat and hamster in vivo and in vitro

[6,7-³H] Estrone (E) and [6,7-³H]estradiol-17 (E₂) have been synthesized by reduction of 6-dehydroestrone and 6-dehydroestradiol with tritium gas. Tritiated E and E₂ were administered by oral gavage to female rats and to male and female hamsters on a dose level of about 300 g/kg (54 mCi/kg). After 8 h, the liver was excised from the rats; liver and kidneys were taken from the hamsters. DNA was purified either directly from an organ homogenate or via chromatin. The radioactivity in the DNA was expressed in the units of the Covalent Binding Index, CBI = (mol chemical bound per mol DNA-P)/(mmol chemical administered per kg b.w.). Rat liver DNA isolated via chromatin exhibited the very low values of 0.08 and 0.09 for E and E₂, respectively. The respective figures in hamster liver were 0.08 and 0.11 in females and 0.21 and 0.18 in the males. DNA isolated from the kidney revealed a detectable radioactivity only in the female, with values of 0.03 and 0.05 for E and E₂, respectively. The values for male hamster kidney were < 0.01 for both hormones. The minute radioactivity detectable in the DNA samples does not represent covalent binding to DNA, however, as indicated by two sets of control experiments. (A) Analysis by HPLC of the nucleosides prepared by enzyme digest of liver DNA isolated directly or via chromatin did not reveal any consistent peak which could have been attributed to a nucleoside-steroid adduct. (B) All DNA radioactivity could be due to protein contaminations, because the specific activity of chromatin protein was determined to be more than 3,000 times higher than of DNA. The high affinity of the hormone to protein was also demonstrated by in vitro incubations, where it could be shown that the specific activity of DNA and protein was essentially proportional to the concentration of radiolabelled hormone in the organ homogenate, regardless of whether the animal was treated or whether the hormone was added in vitro to the homogenate.Carcinogens acting by covalent DNA binding can be classified according to potency on the basis of the Covalent Binding Index. Values of 10³–10⁴ have been found for potent, 10² for moderate, and 1–10 for weak carcinogens. Since estrone is moderately carcinogenic for the kidney of the male hamster, a CBI of about 100 would be expected. The actually measured limit of detection of 0.01 places covalent DNA binding among the highly unlikely mechanisms of action. Similar considerations can be made for the liver where any true covalent DNA binding must be below a level of 0.01. It is concluded that an observable tumor induction by estrone or estradiol is unlikely to be due to DNA binding.Paper presented at the Satellite Symposium of the European Society of Toxicology, Rome, March 29, 1983     

Mechanisms of antiplatelet and antithrombotic activity of midazolam in in vitro and in vivo studies

Dopamine regulates various physiological functions in the central nervous system and the periphery. Dysfunction of the dopamine system is implicated in a wide variety of disorders and behaviors including schizophrenia, addiction, and attention-deficit hyperactivity disorder. Medications that modulate dopamine signaling have therapeutic efficacy on the treatment of these disorders. However, the causes of these disorders and the role of dopamine are still unclear. Studying the dopamine system in a model organism, such as Caenorhabditis elegans, allows the genetic analysis in a simple and well-described nervous system, which may provide new insight into the molecular mechanisms of dopamine signaling. In this review, we summarize recent findings on pharmacological and biochemical properties of the C. elegans dopamine receptors and their physiological role in the control of behavior.     

Extraction and determination of prednisolone in drugs and excipients in ointments and creams and the uniformity of distribution in prednisolone ointment

For the purpose of measuring the contents of prednisolone in low concentrated ointments and creams an instruction was elaborated that includes several steps of extraction, in the resulting solution of which the assay of the steroid by Blue Tetrazolium reaction will be done. The procedure permits the determination of prednisolone in presence of most of usual ingredients of ointment bases except wool alcohols. Also no influence is given by some remedies combined with prednisolone for topical application except coal tar solution. The results confirm a correct reflection of the steroid contents declared respectively the recovery of the steroid added to various ointment bases. Introducing discussion to content uniformity concerning low concentrated ointments is made, and some deviations are shown.     

Penetration and action of edoxudine in vitro and in vivo

The penetration of 5-ethyl-2'-deoxyuridine (edoxudine, Aedurid) from gel base with and without the addition of urea and other adjuvant has been studied in an in vitro model using guinea pig skin. The formulation of 3% edoxudine gel with 5% urea showed the best results. In vivo experiments on hairless mice infected intracutaneously with herpes simplex virus type 1 also showed this formulation's good efficacy as compared to other formulations.     

Pharmacokinetics and tolerability of artesunate and amodiaquine alone and in combination in healthy volunteers

Objectives  The WHO recommends artemisinin-based combination therapies for treatment of uncomplicated falciparum malaria. At least 15 African countries have adopted artesunate plus amodiaquine as treatment policy. As no pharmacokinetic data on this combination have been published to date, we investigated its pharmacokinetic interactions and tolerability in healthy volunteers in Africa. Methods  In a randomized, three-phase, cross-over study, amodiaquine (10 mg/kg) and artesunate (4 mg/kg) were given as single oral doses to 15 healthy volunteers. Artesunate was given to all volunteers on day 0. On day 7 they received either amodiaquine or amodiaquine plus artesunate and the alternative regimen on day 28. The pharmacokinetics of artesunate and amodiaquine and their main active metabolites dihydroartemisinin and desethylamodiaquine were compared following monotherapy and combination therapy using analysis of variance. Results  Thirteen volunteers completed the study, and pharmacokinetic parameters could be determined for twelve volunteers. When given in combination, the mean AUC was lower for dihydroartemisinin [ratio 67% (95% CI 51–88%); P = 0.008] and desethylamodiaquine [ratio 65% (95% CI 46–90%); P = 0.015] when compared with monotherapy. Adverse events of concern occurred in four volunteers (27%): grade 3 transaminitis (n = 1), neutropaenia (n = 2), and hypersensitivity (n = 1). Conclusion  The total drug exposure to both drugs was reduced significantly when they were given in combination. The clinical significance of these interactions is unclear and must be studied in malaria patients. The frequency and nature of adverse events among the healthy volunteers were of concern, and suggest laboratory monitoring would be needed in malaria patients treated with artesunate plus amodiaquine.     

Strain-dependency in motor activity and in concentration and turnover of catecholamines in synchronized rats

Circadian rhythm in motor activity was studied with an Animex motimeter in six strains of rats (ACI, BH, BS, DA, LEW, TNO) synchronized by a 12 hr light: 12 hr dark cycle. ANOVA revealed significant interstrain differences in motor activity as well as in the concentration and turnover of central noradrenaline and dopamine. Strain-dependent differences were also found with regard to tyrosine hydroxylase inhibition on motor activity. However, no significant interstrain correlations were found between endogenous concentration and/or turnover rates of the catecholamines and motor activity in normal and drug-treated rats.     

Comparative in vivo and in vitro studies of phenytoin protein binding and in vitro lipolysis in plasma of pregnant and nonpregnant rats

This investigation was designed to determine the cause of the changes in drug protein binding that occur in rat plasma, particularly in plasma from pregnant animals, during in vitro drug-protein binding measurements. In vivo estimates of phenytoin binding in plasma were obtained from steady-state CSF-plasma concentration ratios in pregnant and nonpregnant rats. Immediate ultrafiltration of heparin- or EDTA-anticoagulated plasma yielded phenytoin free fraction values that were in good agreement with in vivo estimates for nonpregnant rats but that were about one-third higher than in vivo estimates for pregnant animals. In vitro free fraction values tended to increase during incubation of plasma and/or during equilibrium dialysis. The concentrations of the four major endogenous free fatty acids were similar in plasma of pregnant and nonpregnant rats if determined immediately after blood collection. Six hours of incubation at 37 degrees C caused fatty acid concentrations to increase about fivefold and twofold in heparin-anticoagulated plasma from pregnant and nonpregnant animals, respectively. The corresponding increases in EDTA-anticoagulated plasma were only about twofold and 1.14-fold, respectively. These changes were associated with decreased plasma protein binding of phenytoin. The in vivo differences between pregnant and nonpregnant rats with respect to phenytoin binding in plasma are not due to differences in fatty acid concentrations, but the in vitro differences are due primarily to corresponding differences in free fatty acid concentrations if extensive in vitro lipolysis occurs.     

Identification and characterization of in vitro and in vivo metabolites of steroidal alkaloid veratramine

Veratramine, a steroidal alkaloid originating from Veratrum nigrum L., has demonstrated distinct anti‐tumor and anti‐hypertension effects, however, its metabolism has rarely been explored. The objective of the current study was to provide a comprehensive investigation of its metabolic pathways. The in vitro metabolic profiles of veratramine were evaluated by incubating it with liver microsomes and cytosols. The in vivo metabolic profiles in plasma, bile, urine and feces were monitored by UPLC‐MS/MS after oral (20 mg/kg) and i.v. (50 µg/kg) administration in rats. Meanwhile, related P450s inhibitors and recombinant P450s and SULTs were used to identify the isozymes responsible for its metabolism. Eleven metabolites of veratramine, including seven hydroxylated, two sulfated and two glucuronidated metabolites, were characterized. Unlike most alkaloids, the major reactive sites of veratramine were on ring A and B instead of on the amine moiety. CYP2D6 was the major isozyme mediating hydroxylation, and substrate inhibition was observed with a V_max, K_i and Cl_int of 2.05 ± 0.53 nmol/min/mg, 33.08 ± 10.13 µ m and 13.58 ± 1.27 µL/min/mg. SULT2A1, with K_m, V_max and Cl_int values of 19.37 ± 0.87 µ m , 1.51 ± 0.02 nmol/min/mg and 78.19 ± 8.57 µL/min/mg, was identified as the major isozyme contributing to its sulfation. In conclusion, CYP2D6 and SULT2A1 mediating hydroxylation and sulfation were identified as the major biotransformation for veratramine. Copyright © 2015 John Wiley & Sons, Ltd.     

Simultaneous Determination of Sulfation and Glucuronidation of Flavones in FVB Mouse Intestine in Vitro and in Vivo

Glucuronidation and sulfation are the two major phase II metabolic pathways for flavones, natural compounds that hold great potential for improving human health. We investigated the positional preference for sulfation and glucuronidation of seven structurally similar flavones in vitro and in situ. An FVB mouse intestinal perfusion model was used in addition to three small intestine S9 fractions catalyzing sulfation only (Sult enzymes), glucuronidation only (Ugt enzymes) or both (Sult and Ugt enzymes). In both the single and co‐reaction S9 systems, flavones containing 7‐OH groups were conjugated only at 7‐OH despite the presence of other hydroxyl groups, and 7‐OH glucuronidation was faster than sulfation (P < 0.05). The sulfation rate was enhanced in the Sult‐Ugt co‐reaction system, while glucuronidation was usually unchanged by the presence of Sult. In the intestinal perfusate, sulfation patterns were the same in the small intestine and colon, and the excretion rate of 7‐O‐sulfate was the fastest or second fastest. The excretion of 7‐O‐glucuronidates was faster in small intestine (P < 0.05) than in colon. The S9‐mediated sulfation rates of the different flavones were significantly correlated with the excretion rates of the same flavones from perfused intestine. In conclusion, flavone glucuronidation and sulfation rates were sensitive to minor changes in molecular structure. In intestinal S9 fractions, both Ugts and Sults preferentially catalyzed reactions at 7‐OH. The sulfation rate was significantly enhanced by simultaneous glucuronidation, but glucuronidation was unaltered by sulfation. Sulfation rates in mouse S9 fractions correlated with sulfation rates in perfused intestine. Copyright © 2011 John Wiley & Sons, Ltd.     

多廿烷醇与普伐他汀治疗高脂血症的疗效和安全性

目的评价多廿烷醇治疗高脂血症,特别是高胆固醇血症的疗效和安全性。方法多甘烷醇组(试验组,多廿烷醇10mg·d~(-1))和普伐他汀组(对照组,普伐他汀10 mg·d~(-1))各119例。进行随机、双盲、双模拟、阳性药物平行对照试验。观察2组降脂疗效和不良反应发生情况。结果经过12 wk治疗,多廿烷醇组总胆固醇(TC)、低密度脂蛋白胆固醇(LDL)、TC-高密度脂蛋白胆固醇(HDL-C)/ HDL-C、载脂蛋白B_(100)(Apo B_(100))、脂蛋白(Lpa)治疗前分别为(6.6±s0.7)、(4.0±0.6)、(0.10±0.03)、(3.3±0.5)mmol·L~(-1)、(260±184)mg·L~(-1),治疗后分别为(6.0±1.3)、(3.5±0.8)、(O.09±0.04)、(2.7±0.8)mmol·L~(-1)、(130±130)mg·L~(-1),治疗后各指标较治疗前均有非常显著差异(P<0.01)。普伐他汀组TC、LDL-C、TC-HDL-C/HDL-C、Apo B_(100)、Lpa治疗前分别为(6.7±0.8)、(4.1±0.7)、(0.10±0.03)、(3.4±0.5)mmol·L~(-1)、(279±240)mg·L~(-1),治疗后分别为(6.0±1.3)、(3.5±0.9)、(0.09±0.03)、(2.8±0.8)mmol·L~(-1)、(182±213)mg·L~(-1),治疗后各指标较治疗前均有非常显著差异(P<0.01)。但2组相比较,调脂作用相似,无显著差异(P>0.05)。不良反应方面,多廿烷醇组(9.2%)明显少于普伐他汀组(18.5%),2组有显著差异(P<0.05)。不良反应大多较轻微,不需停药能自行缓解。结论多廿烷醇10 mg·d~(-1)降脂效果与普伐他汀10 mg·d~(-1)的疗效相当,均能明显降低TC、LDL-C、TC-HDL-C/ HDL-C、Apo B_(100)、Lpa。多廿烷醇的安全性优于普伐他汀,不良反应轻微,耐受性好。