荧光原位杂交检测乳腺癌人表皮生长因子受体2基因扩增及与临床病理特征的关系

目的:探讨荧光原位杂交法(fluorescence in situ hybridization,HSH)及免疫组织化学法(immunohistochemistry,IHC)检测乳腺癌人表皮生长因子受体2(human epidermal growth factor receptor2,HER-2)的一致性,及HER-2基因扩增与乳腺癌临床病理特征的关系.方法:对北京大学第三医院病理科2008年1月至10月的乳腺浸润性导管癌病例中选取的41例进行HER-2基因扩增的FISH检测和蛋白表达的IHC检测,并对HER-2基因扩增与雌激素受体(estrogen receptor,ER)、组织学分级、癌周脉管内癌栓、腋窝淋巴结转移的关系进行统计学分析.结果:41例乳腺浸润性导管癌中,HER-2基因扩增患者15例,为IHC检测+++者8例,++者6例,+者1例,阴性者0例,分别占IHC+++、++、+及阴性者病例总数的88.89%(8/9)、42.68%(6/14)、8.33%(1/12)及0%(0/6).HER-2基因扩增率在不同组织学分级的乳腺浸润性导管癌中差异无统计学意义(P=0.095).HER-2基因扩增率在ER阴性及弱阳性(+)组高于ER强阳性(++或+++)组(P=0.018),可见脉管内癌栓组高于未见脉管内癌栓组(P=0.000),同侧腋窝淋巴结有转移组高于腋窝淋巴结无转移组(P=0.025).结论:FISH技术可较稳定地应用于乳腺癌HER-2基因扩增的检测,IHC++的患者需进一步行FISH检测;HER-2基因扩增与ER表达呈负相关,与癌周脉管内癌栓、腋窝淋巴结转移呈正相关.     

bcl-2与乳腺癌耐药蛋白基因在弥漫大B细胞淋巴瘤中表达的意义

目的 研究bcl-2和乳腺癌耐药蛋白(BCRP)基因在非霍奇金淋巴瘤(NHL)中的表达及其相关性.方法 对初治弥漫大B细胞淋巴瘤(DLBCL)患者(40例)淋巴结组织液,采用流式细胞术(FCM)及反转录聚合酶链反应(RT-PCR)技术定量检测其BCRP mRNA的表达,同时将患者标本常规石蜡包埋、HE染色和链霉菌素牛物素技术(LSAB)免疫组织化学法标记bcl-2蛋白表达.结果 40例DLBLC患者的bcl-2与BCRP的阳性表达率分别为60.0%(24/40),37.5%(15/40),不同临床分期的DLBCL患者,BCRP阳性表达率差异有统计学意义(x2=6.0606,P<0.05).bcl-2、BCRP表达阳性组有效率低于表达阴性组,差异有统计学意义(x2=5.7618,P<0.05;x2=6.5541,P<0.05);bcl-2和BCRP表达均阳性与均阴性患者的疗效比较,差异无统计学意义(x2=2.0263,P>0.05).结论 BCRP可能在DLBCL的原发多药耐药中发挥重要作用,并有助于化疗疗效的评估及提示疾病转归;bcl-2在DLBCL中的表达对肿瘤恶性程度及预后的判断均有一定意义;联合检测bcl-2和BCRP基因对评价DLBCL预后有较大意义.     

PTEN、MLL基因在T淋巴母细胞淋巴瘤/白血病的表达及其意义

目的 分析肿瘤抑制基因PTEN、混合系白血病(MLL)基因等在T淋巴母细胞淋巴瘤/白血病(T-LBL/ALL)的表达及意义.方法 选用76例T-LBL/ALl患者淋巴结存档蜡块,应用免疫组织化学EnVision法进行 PTEN标记,用20例反应性增生淋巴结标本作正常对照.并用荧光原位杂交(FISH)技术检测MLL基因所在1 1q23染色体的断裂和扩增情况.结果 76例T-LBL/ALL中,PTEN的表达率为64.47%(49/76),低于淋巴结反应性增生的100%(20/20)(λ2=19.220,P＜0.05).PTEN表达与临床分期、Ki-67、乳酸脱氢酶(LDH)呈负相关(P＜0.05).76例T-LBL/ALL中,MLL基因发生1 1q23染色体断裂13例(17.11%),扩增18例(23.68%).MLL基因断裂组总体生存率(25.0%)低于非断裂组(43.6%)(λ2=11.357,P＜0.05).MLL基因扩增组总体生存率(17.1%)低于非扩增组(42.7%)(λ2=4.533,P＜0.05).结论 抑癌基因PTEN表达降低在T-LBL/ALL的发生发展中可能具有重要作用.MLL基因发生染色体1 1q23断裂和扩增有助于对T-LBL/ALL预后的判断,发生MLL基因断裂或扩增的T-LBL/ALL预后较差,提示MLL基因断裂或扩增可能为T-LBL/ALL的一种分子亚型.     

非特指型外周T细胞淋巴瘤FGR和TP73基因改变的研究

目的 研究非特指型外周T细胞淋巴瘤(PTCL-NOS)中FGR和TP73的基因改变,验证前期基于芯片的比较基因组杂交(a-CGH)研究结果,为PTCL-NOS的发生、发展提供科学依据.方法 间期双色荧光原位杂交(FISH)技术检测34例PTCL-NOS(其中19例经过a-CGH检测)中FGR和TP73的基因改变,所用探针为缺口平移法自制的FGR和TP73特异位点探针以及商品化1号染色体着丝粒探针(CEP1).结果 34例中出现基因改变者7例(20.6%),其中FGR扩增1例,TP73扩增2例;FGR和TP73同时出现基因改变4例(11.8%),其中同时杂合性缺失(LOH)1例,同时扩增3例.CEP1扩增4例(11.8%),与TP73/FGR基因扩增同时存在.Kaplan-Meier生存分析显示基因改变组较基因无改变组以及TP73改变组较TP73无改变组均有预后不良的趋势,但差异无统计学意义(均P>0.05).结论 FISH结果部分验证了前期a-CGH的研究发现;淋巴瘤相关基因FGR和TP73改变以及1号染色体多倍体可能在PTCL-NOS的发生、发展中起着重要的作用.     

信号转导及转录活化因子3蛋白在乳腺癌组织中的表达及其临床意义

目的:检测人信号转导及转录活化因子3(STAT3)蛋白在乳腺癌组织中的表达,探讨其与乳腺癌患者临床病理参数及预后的关系.方法:应用免疫组织化学SP法检测67例乳腺癌及其配对正常乳腺组织中STAT3蛋白的表达水平,分析具有不同临床病理参数患者间STAT3的表达情况.结果:STAT3蛋白在乳腺癌组织的表达水平明显高于正常乳腺组织(P<0.001),STAT3表达水平与乳腺癌患者年龄、肿瘤大小和组织学类型无关联性(P>0.05),与乳腺癌临床分期、淋巴结转移情况及患者生存时间有关联(P<0.05).临床分期越高的乳腺癌组织中STAT3的表达阳性率越高(P<0.05);有淋巴结转移者STAT3阳性表达率明显高于无淋巴结转移者(P<0.05);STAT3阳性表达的患者生存时间[(51.1±4.4)月]明显短于阴性表达的患者[(86.0±4.9)月](P<0.05).结论:STAT3蛋白的过表达可能在乳腺癌的发生发展过程中发挥了促进作用,并且是乳腺癌患者预后不良的一个重要指标.     

儿童侵袭性成熟B细胞淋巴瘤的临床病理学及分子遗传学特点

目的 分析总结中国儿童各类型侵袭性成熟B细胞淋巴瘤的临床病理学及分子遗传学特点,为其诊断的标准化提供依据.方法 收集97例儿童侵袭性成熟B细胞淋巴瘤石蜡包埋组织标本,包括伯基特淋巴瘤(BL)81例、弥漫大B细胞淋巴瘤(DLBCL)8例、介于BL和DLBCL间的不能分类的B细胞淋巴瘤(BL/DLBCL)8例,利用免疫组织化学技术和间期荧光原位杂交(FISH)技术检测其免疫表型和分子遗传学特征.结果 BL的bcl-2和MUM1的阳性率分别为3%(2/66)和17%(12/71),DLBCL分别为50%(4/8)和63%(5/8),BL/DLBCL分别为50%(4/8)和63%(5/8).BL、DLBCL和BL/DLBCL的Ki-67平均值分别为(93±4.4)%、(83±14.3)%和(80±11.5)%.BL、DLBCL和BL/DLBCL的c-myc基因易位的比例分别为98%(79/81)、38%(3/8)和50%(4/8).38%(3/8)的DLBCL和25%(2/8)的BL/DLBCL存在bcl-6基因的多拷贝,BL与DLBCL之间、BL与BL/DLBCL之间bcl-2、MUM1和Ki-67平均值的差异及c-myc基因易位和bcl-6基因多拷贝的差异均有统计学意义(均P＜0.05).结论 儿童侵袭性成熟B细胞淋巴瘤的诊断和分型需要综合分析形态学、免疫表型和分子遗传学特征.儿童BL/DLBCL可能是DLBCL的一个亚型.CD10+、bcl-6+、bcl-2-、Ki-67＞90%、伴有IGH/c-myc重排、不伴有bcl-2和bcl-6重排时,支持BL的诊断;bcl-2+、Ki-67为50%～90%,同时伴有bcl-6基因的多拷贝时,支持DLBCL或BL/DLBCL的诊断.     

阴道镜诊断宫颈病变52例分析

目的:探讨阴道镜对宫颈病变的诊断价值.方法:将我院2009年6月至2010年6月因宫颈病变就诊的52例,该组患者均采用阴道镜检查及宫颈活检,分析阴道镜检查及活检结果,评定该检测方法的临床价值.结果:该组患者中以慢性宫颈炎最多,占76.9%(40/52).阴道镜检查阴性病例44例,组织病理学检查阴性病例43例,诊断符合率为97.7%(43/44).且两者的阳性诊断符合率为88.9% (8/9).结论:阴道镜检查技术方法简单方便,且准确率较高,有较好的临床应用价值.     

中国人经典霍奇金淋巴瘤中人类白细胞共同抗原表达的研究

目的 既往的研究发现在高加索人中,经典霍奇金淋巴瘤肿瘤细胞人类白细胞共同抗原(HLA)的表达与EB病毒感染密切相关,研究在亚洲人中两者的相关性.方法 随机选取北京大学医学部病理学系常规外检及会诊病例中确诊为经典霍奇金淋巴瘤的145例,所有病例均有石蜡包埋组织蜡块.常规HE染色、形态学观察,根据WHO分类标准对所有病例进行重新分类.原位杂交方法检测EB病毒编码的小RNA(EBER)以提示肿瘤与EB病毒的相关性.HLA-Ⅰ类抗原的表达使用HC-10和β 2-微球蛋白抗体检测,而HLA-Ⅱ类抗原的表达使用CR3/43抗体检测.结果 145例中,40%(58例)的病例为EB病毒相关性.EB病毒阳性病例中,混合细胞型较结节硬化型更为常见(71%比16%,P＜0.001).HLA-Ⅰ类抗原在EB病毒阳性病例中的表达率明显高于EB病毒阴性的病例(79%比30%,P＜0.001).而HLA-Ⅱ类抗原的阳性率在EB病毒阳性和阴性病例中差异无统计学意义(52%比43%,P=0.277).结论 中国人经典霍奇金淋巴瘤HLA-Ⅰ类抗原的表达与EB病毒感染密切相关,与高加索人一样,但HLA-Ⅱ类抗原的表达与EB病毒感染无显著相关性.     

rAAV/CEA转染树突状细胞诱导特异性CTL杀伤MCF-7细胞系CD44+ CD24-/low乳腺癌干细胞

目的:携带癌胚抗原(carcinocmbryonic antigen,CEA)基因的重组人腺相关病毒(reconstructive human adeno-association virus,rh-AAV)感染树突状细胞(dendritic cell,DC)诱导获得抗原特异性细胞毒性T淋巴细胞(cytotoxic lymphocyte,CTL),体外检测其对CD44+CD24-/low乳腺癌干细胞的杀伤效果.方法:分离供者外周血单个核细胞,以细胞因子白介素-4(interleukin-4,IL-4)、粒细胞-巨噬细胞克隆刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)和肿瘤坏死因子α(tumor necrosis factor-alpha,TNF-α)诱导培养获得DC,细胞因子白介素-2(interleukin-2,IL-2),刺激培养获得T淋巴细胞.含CEA基因的rh-AAV感染培养DC,诱导成熟后将DC和T细胞混合培养获得CTL细胞.流式分选MCF-7和MA-MDB-231细胞系中CD44+CD24-/low乳腺癌干细胞,MTT法检测CTL细胞对CD44+CD24-/low乳腺癌干细胞的杀伤效果.结果:MCF-7和MDA-MB-231中CD44+/CD24-/low群比例分别为5.1%和76.3%.CEA基因转染DC诱导的CTL细胞对表达CEA抗原的MCF-7杀伤率为46.5%±15.0%,与未转染组相比,差异有统计学意义(P=0.009);对MCF-7细胞中分选的CD44+CD24-/low乳腺癌干细胞的杀伤率为44.7%±28.2%,明显高于未转染组.对非乳腺癌干细胞杀伤率为50.6%±22.2%,与未转染组相比,差异有统计学意义(P=0.05).在不表达CEA基因的MDA-MB-231乳腺癌细胞,CEA转染诱导的CTL细胞对未分选细胞、分选的CD44+/CD24-/low亚群和非干细胞亚群的杀伤率与未转染的对照组相比,差异均无统计学意义(P＞0.05).结论:携带CEA基因rh-AAV转染DC诱导产生的抗原特异性CTL细胞可杀伤表达CEA的乳腺癌细胞,对CD44+CD24-/low乳腺癌干细胞也具有一定的杀伤活性,提示免疫治疗可能是治疗乳腺癌干细胞潜在有效的手段.     

环氧合酶-2抑制剂对白血病细胞HL-60的化疗增敏作用及机制

目的 观察环氧合酶-2(COX-2)抑制剂塞来昔布对白血病细胞株HL-60的化疗增敏作用,并对其机制进行初步探讨.方法 MTT法评估塞来昔布、多柔比星及二者联合对HL-60细胞的生长抑制效应;流式细胞术(FCM)检测细胞的凋亡;反转录聚合酶链反应(RT-PCR)检测Survivin基因的表达;Western blotting检测Survivin蛋白的表达.结果 多柔比星联合塞来昔布5和10μmol/L对HL-60细胞的半数抑制浓度(IC50)分别为0.25及0.16μg/ml,明显低于多柔比星单用的IC50(0.48μg/ml);多柔比星0.10μg/ml联合10 μmol/L塞来昔布下调Survivin基因mRNA及蛋白的表达;联合塞来昔布5和10 μmol/L的凋亡率[分别为(13.07±1.66)%及(22.36±1.84)%]较多柔比星0.10μg/ml单用[(5.72±1.25)%]明显增加(P<0.01).结论 COX-2抑制剂塞来昔布对白血病细胞株HL-60具有明显的化疗增敏作用,其初步机制涉及下调Survivin的表达,增加细胞凋亡.     

Causes of death and incidence of cancer in physicians and lawyers in Iceland

A retrospective study is accomplished in Iceland to study whether mortality and cancer incidence among male physicians (1,210) were lower than those among men of the general population and lawyers (1,032). Overall mortality among lawyers was similar to that of the general male population, however, mortality among the physicians was lower than that of the general population and the lawyers, due to lower mortality for all cancers (SMR 0.73), cerebrovascular diseases (SMR 0.53) and respiratory diseases (SMR 0.54). The physicians had higher mortality for suicide committed by drugs, solid or liquid substances. Cancer was not as frequent among the physicians as among the lawyers, particularly for lung cancer, the SIR was 0.45, but the rates were higher for cancer of the colon and brain among the physicians than among others.     

Pharmacokinetics and metabolism of vinyl fluoride in vivo and in vitro

Vinyl fluoride (VF) is an inhalation carcinogen at concentrations of 25 ppm or greater in rats and mice. The main neoplastic lesion induced in rodents was hepatic hemangiosarcomas, and mice were more sensitive than rats. In a first set of experiments, groups of three rats or five mice were exposed to VF in a closed-chamber gas uptake system at starting concentrations ranging from 50 to 250 ppm. Chamber concentrations of VF were measured every 10-12 min by gas chromatography. Partition coefficients were determined by the vial equilibration technique and used as parameters for a physiologically based pharmacokinetic (PBPK) model. Mice showed a higher whole-body metabolic capacity compared to rats (Vmax = 0.3 vs 0.1 mg/hr-kg). Both species had an estimated Km of < or = 0.02 mg/liter. The specificity for the oxidation of VF in vivo was determined by selective inhibition or induction of CYP 2E1. Inhibition with 4-methylpyrazole completely impaired VF uptake in rats and mice, whereas induction with ethanol (rats only) increased the metabolic capacity by two- to threefold. The pharmacokinetics of VF were also investigated in vitro. Microsomes from rat and mouse liver were incubated in a sealed vial with VF and an NADPH-regenerating system. Headspace concentrations (10-300 ppm) were monitored over time by gas chromatography. Consistent with the in vivo data, VF was metabolized faster by mouse microsomes than by rat microsomes (Vmax = 3.5 and 1.1 nmol/hr-mg protein, respectively). The rates of metabolism by human liver microsomes were generally in the same range as those found with rat liver microsomes (Vmax = 0.5-1.3 nmol/hr-mg protein), but one sample was similar to mice (Vmax = 3.3 nmol/ hr-mg protein). Metabolic rates in human microsomes were found to correlate with the amount of CYP 2E1 as determined by Western blotting and by chlorzoxazone 6-hydroxylation. It is concluded that the greater metabolic capacity of mice for VF both in vivo and in vitro may contribute to their greater susceptibility to tumor formation. CYP 2E1 is clearly the main isozyme involved in the oxidation of VF in all species tested. VF pharmacokinetics and metabolism in humans may depend upon the interindividual variability in the expression level of CYP 2E1. The excellent correspondence between in vivo and in vitro kinetics in rodents improves. substantially the degree of confidence for human in vivo predictions from in vitro data.     

Development and maintenance of bile canaliculi in vitro and in vivo

Four types of high-flux hemodialyzers, Primus 2000 (high-flux polysulfone 2.0 m2), Altra-Flux 170 G (cellulose diacetate 1.7 m2), FLX-15 GW (polyester-polymer alloy 1.5 m2) and PAN-85 DX (polyacrylonitrile 1.7 m2) were evaluated in vivo. A total of 12 stable chronic hemodialysis patients participated in the study and each type of dialyzer was tested once in 9 of them. Blood samples for the measurement of BUN, creatinine, phosphate, uric acid, albumin and beta2-microglobulin (beta2M) were drawn before and 5 min after the end of the study dialysis. During dialysis, which was performed in all patients with a blood flow rate of 250 ml/min for 240 min, the dialysate (550-600 ml/min) was collected every hour and samples were drawn for the measurements of all the above substances. The mean total amount of low-molecular substances removed per session by each dialyzer was very close to 19.5 g for urea, 2.0 g for creatinine, 0.9 g for phosphate and 1 g for uric acid. The one-third (30-33%) of the above amounts were removed during the first hour of dialysis. Dialyzers' clearances for creatinine and uric acid were significantly higher in Primus dialyzer comparing to FLX-15 GW (p < 0.05) while the clearance for urea showed a borderline significance (p = 0.055). No difference was found either among Altra-Flux 170 G, FLX-15 GW and PAN-85 DX or between Primus and PAN-85 DX dialyzers. Phosphate clearance did not show any difference among the four dialyzers. The lowest amount of albumin removed per session was 0.75 g by PAN-85 DX and the highest 1.8 g by FLX-15 GW, while the equivalents for beta2M were 80 mg by Altra-Flux 170 G and 142 mg by PAN-85 DX. A significant adsorption of beta2M on these dialysis membranes was indicated by the combination of a satisfactory serum beta2M reduction ratio (post-/predialysis values = 0.52, 0.77, 0.60, 0.55) with a reduced beta2M clearance (23.9, 13.6, 20.2, 25.1 ml/min). During the first hour of dialysis, in comparison to the following time, the highest amounts of albumin and beta2M (expressed as percentage of total) were removed by the Primus 2000 dialyzer. Our results indicate that under conventional conditions small differences in the surface area of the high-flux dialyzers are unimportant regarding the removal of low molecules. However, the composition of the membrane seems to play an important role in the removal of high-molecular substances.     

Smoking and pursuit of thinness in schoolgirls in London and Ottawa

It has been proposed that teenage girls often smoke cigarettes to protect themselves from the impulse to binge eat, with its feared weight-gain consequences, particularly when other measures such as greater dietary restraint have failed. The present study looked at the relationship between body mass index and standardised questionnaire responses concerning smoking, alcohol consumption, moods, weight changes, attitudes to body weight and shape, dietary patterns and menstruation in 1936 British (London) and 832 Canadian (Ottawa) schoolgirls. Data analysis revealed links between cigarette smoking and body weight/shape concerns, and awareness by subjects of these links; there was also a tendency for smokers in these two populations to be overweight but not grossly obese. Smoking was also related at all ages to being postmenarchal. The London population in particular revealed an association between smoking and a weight loss of 7 kg or more at some stage since puberty. Smoking was also linked, in a minority, with regular vomiting undertaken as a further defence against weight gain when overeating had occurred. These associations existed alongside the major and predictable association found between alcohol consumption and smoking. Similarities between the British and Canadian schoolgirls were striking in respect of rank order of reasons given for smoking and consequences of giving it up. Since smoking amongst older women is reportedly associated with below-average body weight it may indeed be effective in helping to curb weight gain. Our study provided little evidence of association between smoking and generalised anxiety or social anxiety (in either population), or depression (in the British cohort). We suggest that any preventive psychological approach to teenage female smoking should include attention to weight gain anxiety and consequent pursuit of thinness.     

Alterations in metabolism of H-epinephrine in the pregnant and non-pregnant state and in males

The influence of sex, pregnancy and parturition of biological accumulation and metabolic fate was studied in Sherman rats. 3H-epinephrine was used as a tracer to determine differences in metabolite formation in normal males and females as well as pregnant rats from 18 days post coitum to the end of parturition. The disappearance of 3H-epinephrine and its metabolites was measured in blood, heart, kidney and brain at different intervals but 20 min after the injection of tracer was found the most appropriate time to find appreciable radioactivity in most of the organs studied. All the comparisons for 3H-epinephrine accumulation and its transformation to metabolites have been reported 20 min post-perfusion period. Marked changes of high statistical significance in 3H-epinephrine accumulation and its transformation to 3H-metanephrine and 3H-acid metabolities were observed between males and females. Heart, adrenals, and spleen showed lower rate of metabolism but higher rate of accumulation during pregnancy. Kidney, ovary, and uterus demonstrated higher rate of metabolism but lower rate of accumulation during gestation. In brain regions, hypophysis discriminated greatly between males and females for the three parameters studied. There were important alterations in metabolite formation during 18 and 21 days of pregnancy. The observed variations have been considered due to modified endocrine activity during pregnancy and parturition.     

Comparative binding and toxicity of saxitoxin and saxitoxinol in mice and in cultured cells

The binding characteristics of saxitoxin (STX), a known voltage-gated sodium channel blocker, and its analog saxitoxinol (STXOL), were studied in neuroblastoma, peritoneal macrophage, hepatocytes and PC-12 cell lines. 3H-STXOL bound to the cell-surface sites which appear to be the same as those occupied by 3H-STX and which can, therefore, be identified as STX receptors. The relative agreement of respective Kd obtained by saturation, competition, association and dissociation kinetics for STX and STXOL suggest the absence of any artifact in binding measurements. Unlike STX, STXOL was non-toxic to mice by intratracheal instillation. The major advantage of using 3H-STXOL is that the tritium label is not exchangeable. Data from this study suggest that 3H-STXOL can be used to identify STX receptors at 37 degrees C.     

Response of Plasmodium falciparum to chloroquine and Fansidar in vivo and chloroquine and amodiaquine in vitro in Uganda

The response of P. falciparum to chloroquine and pyrimethamine-sulfadoxine in vivo and chloroquine and amodiaquine in vitro was investigated in parasitaemic school children from six locations. Mean parasite sensitivity to chloroquine at day 7 was 74% (range 61-97) with parasite clearance rates between 2-3 days and complete defervescence in 85% of febrile children. Sensitivity declined in the four sites followed up to day 14 to 45% (range 37-53). Parasites were significantly more sensitive to pyrimethamine/sulfadoxine at 5/6 sites (100% day 7) but 5% of subjects became parasitaemic by day 14. In vitro isolates were significantly less sensitive to chloroquine than to amodiaquine with a mean 99% effective concentration of 348 mumol/L compared to 6.44 mumol/L. Clearly the role of chloroquine as the primary therapy for uncomplicated P. falciparum malaria should be reconsidered especially in the light of increasing disease severity and resurgence. Amodiaquine may be suitable alternative with pyrimethamine/sulfadoxine as second line and for more severe malaria prior to referral. The cost of alternative antimalarials and the dynamic and deteriorating pattern of resistance are powerful arguments for more objective slide diagnosis to minimise drug pressure and a regular drug sensitivity surveillance system. We believe that the latter should concentrate on measuring clinical drug efficacy in symptomatic outpatients rather than in asymptomatic children while the former needs more pragmatic and economical strategies possibly centred on seasonality and risk.     

The prognostic and diagnostic value of total estriol in urine and in serum and of human placental lactogen hormone in serum in the last part of pregnancy

The benefit to the antenatal care from using estriol and human placental lactogen (hPL) determinations is emphasized and likewise that special attention must be paid to values below the reference intervals. Referring to biosynthesis and place of production, a survey is given of the various causes of low estriol and/or low hPL values, and a number of clinical cases illustrate the matter. A follow-up study of children from pregnancies with low estriol values has disclosed an alarming number of cases of severe handicaps.     

Residues of perchlorethylene and petroleum in rendered fats and in feeding meals

Methods were developed for the demonstration and determination of perchlorethylene and petrol in rendered fats and meals for feeding purposes. The methods are based on the collection and the vapours of extracting media over samples of rendering-plant products and on their identification and determination by gas chromatography. Under the conditions of the method, the limit of perchlorethylene extractibility is 0.05% and the limit of petrol extractibility is 0.01% (weight). Under adapted conditions the two limits can be reduced. The reproducibility of the results at practically occuring concentrations of residues in products is characterized by the value of the variation coefficient which reaches 2.79 in perchlorethylene and 6.05 in petrol. The maximum duration of the analysis of one sample is five minutes. In all media have been demonstrated to be present. Their content was high and considerably variable in the fats from rendering plants; in meat-bone meals from petrol extraction the content of residues reached only trace values. It is recommended to use the presented analytic methods for thorough and regular objective control of the residues of extracting media in the products of rendering plants for feeding purposes.