肝细胞癌中SEMA3B基因的表达及其意义

目的 通过对SEMA3B基因在原发性肝细胞癌（HCC）组织中表达的检测，探讨该基因与HCC发生机制的关系。方法 采用逆转录-聚合酶链反应（RT—PCR）方法检测35例HCC组织及癌旁肝组织SEMA3BmRNA表达水平。结果 （1）在所有受检的癌旁肝组织中，SEMA3BmRNA表达率为85．7％（30／35），明显高于癌组织的表达率20．0％（7／35），差异有统计学意义（P〈0．01）；（2）合并有肝硬化（20／28）或者乙肝（19／28）的患者，其癌组织中SEMA3B基因的表达缺失率显著高于无肝硬化（8／28）或乙肝阴性（9／28）患者（P〈0．05）；（3）SEMA3B基因的异常表达情况与年龄、性别的差异无统计学意义（P〉0．05），与HCC患者术前肝功能Child—Pugh分级评价和血清AFP的检测值高低无相关（P〉0．05）；（4）SEMA3B的表达缺失率与TNM分期关系无统计学差异，低分化型癌组织中SEMA3B基因缺失率高于中、高分化型，但差异无统计学意义（P〉0．05）。此外，该基因的缺失与肿瘤大小无关（P〉0．05）。结论 SEMA3B基因在HCC组织中的高频表达缺失说明该基因的失活与HCC的发生密切相关。SEMA3B基因是定位于染色体3p21．3区域的与HCC相关的抑癌基因，并且可能在HCC的发生发展方面起重要作用。     

抑癌基因PTEN和p16在肝细胞性肝癌中的表达及其意义

目的探讨PTKN和p16基因在肝细胞性肝癌（HCC）中的表达及其意义。方法采用免疫组化技术S—P法检测PTKN蛋白及p16蛋白在43例HCC及其相应癌旁组织中的表达。结果PTKN蛋白在HCC组织中的阴性表达率明显高于在癌旁组织中的阴性表达率（P〈0．01）;PTKN蛋白表达阴性率与HCC的分化程度、临床分期呈负相关,与AFP甲值及有无转移、HBV感染相关,与p16蛋白缺失率呈正相关,而与患者年龄、性别、肿瘤直径无关。比较PTKN阴性表达和阳性表达的HCC患者的术后1,3,5年生存率,差异在统计学上有显著性意义。结论PIEN失活部分参与了HCC的发生、发展进程。PIEN基因、p16基因的失活及HBV感染可能在HCC的发生、发展中存在协同关系。PTEN蛋白表达的检测可为HCC患者术后的预后判断提供参考。     

Shh和Ptch在肝细胞癌中的表达研究

目的探讨肝细胞癌（HCC）中Shh和Ptch的表达及其意义。方法应用组织芯片技术、免疫组化和原位杂交方法检测100例HCC、25例癌旁组织和5例正常肝组织中Shh和Ptch的表达情况。结果免疫组化检测Hcc中Shh和Ptch的阳性表达率分别为19．28％（16／83）、24．68％（19／77）;原位杂交检测结果分别为44．29％（31／70）、15．85％（13／82）;两种基因在HCC中的表达相关,均与非肿瘤组织的表达有显著性差异,并分别与肿瘤的大小及分化程度相关。结论Shh和Ptch的表达参与了HCC的发生,它们可能是HCC治疗的理想靶标。     

p14ARF蛋白在人肝细胞癌中的表达缺失及其临床意义

目的研究人肝细胞癌中p14ARF蛋白的表达缺失情况及其与临床病理的关系。方法采用链霉菌素-生物素（SP）免疫组织化学法及免疫印记反应（Western blot）对45例肝细胞癌组织、39例癌旁组织及37例非肿瘤肝组织标本进行检测分析。结果p14ARF蛋白在肝癌肿瘤组织中表达缺失率为84．496（38／45），p14ARF蛋白Ⅰ级-Ⅱ级（高分化）病例p14ARF蛋白缺失率为44．496，Ⅲ级-Ⅳ级（低分化）病例p14ARF蛋白缺失率为94．496（P〈0．05）。结论p14ARF蛋白表达缺失与肝细胞癌的发生、发展有一定相关性。提示p14ARF基因的失活可能是肝细胞癌恶性演变的重要因素。     

NF-кB与P-gp在肝细胞肝癌组织中的表达

目的探讨核转录因子（NF-κB）与P糖蛋白（P-gp）在肝细胞肝癌（HCC）及癌旁肝组织的表达及其相互关系。方法采用免疫组织化学S-P法检测30例HCC中NF-κB、P-gp的表达情况，以30例相应癌旁肝组织作为对照组，所得结果用统计学卡方检验及列联表等相关分析进行比较。结果NF-κB蛋白在HCC、癌旁肝组织中阳性表达率分别为63．3％（19／30）、16．7％（5／30），HCC与癌旁肝组织中NF-κB蛋白表达差异有显著性（P〈0.05）；P-gp蛋白在HCC、癌旁肝组织中阳性表达率分别为83．3oA（25／30）、30％（9／30），HCC与癌旁肝组织表达差异有显著性（P〈0.05）。NF-κB蛋白的表达与P-gp蛋白的表达呈正相关（r=0.89，P〈0.05）。结论NF-κB和P-gp在HCC中的表达提示其可能参加了HCC和多药耐药的发生。NF-κB与P-gp在HCC表达中呈正相关，提示NF-κB与P-gp可能介导的多药耐药，共同参与了HCC多药耐药机制。     

乳腺癌p16，p53基因蛋白和mRNA的表达及其意义

目的:探讨乳腺癌组织p16和p53基因蛋白及其mRNA的表达及其意义。方法:采用免疫组化法和RT-PCR技术，检测乳腺癌组织和癌旁组织中p16和p53基因蛋白及其mRNA的表达。分析其表达与乳腺癌临床病理学特征之间的关系。结果:癌旁乳腺组织中p16基因蛋白的表达率为90.0%（27/30），p53基因蛋白的表达率为6.67%（2/30）;乳腺癌组织中p16基因蛋白表达率为38.3%（23/60），p53基因蛋白表达率为48.3%（29/60）。癌旁乳腺组织的p16 mRNA显著高于乳腺癌组织(P=0.023)，而在乳腺癌组织的p53mRNA水平显著高于癌旁乳腺组织（P=0.001）。乳腺癌细胞分化程度越高p16蛋白表达率越高，而p53表达蛋白正相反;有淋巴结和/或器官转移者p16蛋白表达率明显低于无淋巴结和/或器官转移者，而p53蛋白的表达率则相反。p16mRNA在组织学Ⅰ级的水平显著高于Ⅱ，Ⅲ级，而Ⅱ，Ⅲ级之间差异无显著性;p53mRNA在组织学Ⅰ级的水平显著低于Ⅲ级。有淋巴结和/或器官转移的p16mRNA和p53mRNA表达明显低于和高于无淋巴结和/或器官转移者。结论:p16和p53基因的改变与乳腺癌的发生发展关系密切，可用于评估乳腺癌的预后。     

CT9和CT10基因在肝细胞癌中的表达及临床意义

目的检测肿瘤睾丸抗原CT9和CT10基因mRNA在肝细胞癌（HCC）中的表达情况。方法用逆转录-聚合酶链反应（RT-PCR）方法对HCC病人的癌组织和相应癌旁组织及对照（肝硬化和正常肝组织）中CT9和CT10mRNA的表达进行了检测,随机选取每一种抗原RT-PCR呈阳性的产物4例进行DNA序列测定,结合AFP、抗HCV、HgsAg、肿瘤直径、分化程度等临床指标进行分析。结果56例HCC病人中,CY9、CT10基因mRNA阳性率分别是51．8％（29／56）和44．6％（25／56）,至少表达一种CTA者达66．1％（37／56）,表达两种者为30．4％（17／56）;电泳显示PCR产物与阳性对照大小一致;而癌旁组织中仅有CT9基因nRNA表达为阳性,阳性率为7．1％（4／56）,但表达强度明显弱于在癌组织中的表达,进一步的病理检查未发现CY9表达呈阳性的癌旁组织中有癌细胞。所测10例肝硬化和10例正常肝组织均未检测到CT9和CT10的表达。DNA测序结果表明RT-PCR产物确为CT9和CT10cDNA。CT9和CT10的表达与年龄、性别、肿瘤大小、分化程度、AFP水平、HBV和HCV感染无显著相关性（P〉O．05）。而在部分AFP正常（〈25ng／L）HCC病人中存在CT9和CT10基因的表达。结论CT9和CT10基因在HCC中呈高比例、高特异表达,为HCC免疫治疗提供了新的理想靶位;CT9和CT10同时在HCC中表达,为应用以这些肿瘤抗原为基础的多价瘤苗治疗HCC提供了实验依据。     

3p21.3区域相关抑癌基因——BLU在肝癌组织中的表达及临床意义

目的探讨3p21．3区域相关抑癌基因——BLU在肝癌组织中的表达及临床意义。方法采用RT—PCR的方法检测BLU基因在39例肝癌组织以及相应的癌旁组织中的表达。结果35．9％（14／39）癌组织表达,87．2％（34／39）癌旁组织表达,两者差异有统计学意义（P〈0．01）;合并有肝硬化或者乙肝表面抗原阳性的患者,其肝癌组织中BLU基因的表达失活的比例显著高于无肝硬化或乙肝表面抗原阴性患者（P〈0．01）。低分化肝癌较中、高分化肝癌BLU基因未表达的比例显著增高（P〈0．01）。肝癌组织中BLU的表达情况与年龄、性别、术前肝功能Child—Pugh分级及肿瘤大小的差异无统计学意义（P〉0．05）。结论BLU基因可能参与调控肝癌的发生、发展过程,并进而影响其生物学特点及预后。     

抑癌基因PTEN在肝细胞癌中的表达及其意义

目的：探讨肝细胞癌（HCC）组织中抑癌基因PTEN蛋白的表达及其意义。方法：应用免疫组织化学SP法，检测53例HCC组织和癌周组织中PTEN蛋白的表达水平。结果：53例HCC中PTEN蛋白阳性率（52．8％）和阳性强度明显低于癌旁组织（92．5％）（P〈0．01，P〈0．05）；HCC组织中PTEN蛋白的表达与HCC的分化程度密切相关（P〈0．05），HCC分化愈差，PTEN蛋白表达愈弱；PTEN蛋白表达还与癌拴形成有关（P〈0．005）。结论：PTEN的异常表达在HCC发生、发展过程中可能起重要作用，其表达水平有可能作为反映HCC进展和预后的生物学指标。     

肝细胞癌组织中p16,p27蛋白的表达及其意义

为探讨p16,p2 7蛋白在肝细胞癌 (HCC )中的表达情况、相互关系及其临床病理学意义。笔者应用免疫组织化学方法 (SABC法 ) ,检测 49例HCC组织及癌旁组织中 p16,p2 7蛋白表达水平。结果示 49例HCC中 ,p16,p2 7蛋白阳性表达率分别为 3 6.73 %和 40 .82 % ;而癌旁组织阳性表达率为89 .80 %和 91.84%。p16蛋白表达与HCC的分化程度密切相关 (P <0 .0 5 ) ,高分化I～II级中p16阳性率为 5 5 .5 6% ,显著高于III～IV级的 2 5 .81%。p2 7蛋白表达与HCC有无淋巴结、远处转移有显著相关性 (P <0 .0 1)。提示 p16与p2 7蛋白的表达缺失在HCC发生中可能起重要作用。p2 7蛋白可作为衡量HCC预后的有用指标。     

HYPNOTIC AND ANAESTHETIC ACTION OF THIOPENTONE AND MIDAZOLAM ALONE AND IN COMBINATION

This study examined the interaction between i.v. administeredmidazolam and thiopentone on the loss of response to verbalcommand ("hypnosis") and the loss of response to transcutaneouselectrical stimulation of the ulnar nerve ("anaesthesia") inpatients presenting for minor elective surgery. Dose-responsecurves for thiopentone and midazolam individually and in combinationwere determined using the two end-points in 300 unpremedicatedpatients. For hypnosis a highly significant (P < 0.001) supraaddictive(synergistic) interaction was found, the combination having1.8 times the expected potency of the individual agents. Althoughmidazolam failed to produce anaesthesia in the dose range used,the dose of thiopentone required to produce anaesthesia wasreduced by 50% in the presence of midazolam. The mechanism ofinteraction and the potential role of benzodiazepine-barbituratecombinations are discussed and the observed synergistic anaesthesiainteraction is used to explain the potentially dangerous combinationof benzodiazepines with other potent CNS depressants such asbarbiturates and alcohol. ^*Present address: Prince of Wales Hospital, Shatin, Hong Kong.     

THIOPENTONE AND ETOMIDATE CONCENTRATIONS IN MATERNAL AND UMBILICAL PLASMA, AND IN COLOSTRUM

We have measured concentrations of etomidate and thiopentonein maternal plasma, umbilical venous plasma and colostrum afterinduction of anaesthesia in 40 patients undergoing Caesareansection. Mean plasma etomidate concentration declined rapidly(1242.0 ng ml^–1 at 5 min, 434.0 ng mt1 at 15 min, 64.2ng ml^–1 at 30 min, 7.0 ng ml^–1 at 60 min and undetectable2 h after the injection). Mean plasma concentrations of thiopentonedeclined more slowly (6.09 µg ml^–1 at 5 min, 2.64µg ml^–1 at 2 h, 1.35 ng ml^–1 at 4 h, 0.86ng ml^–1 at 9 h and 0.59 ng ml^–1 at 12.h). Mean umbilicalvenous thiopentone concentration was 4.72 µg ml^–1.whereas the thiopentone concentration in the maternal sampleat 5 min was 6.09 g ml^–1, giving an umbilical.maternalvein ratio of 1:1.3. Mean umbilical etomidate concentrationwas 51.7 ng ml^–1 and the corresponding maternal vein sample(5 min) was 1242.0 ng ml^–1 (P < 0.001), giving an umbilical:maternal vein ratio of 1:24. Mean concetrations of thioptonein colostrum were 1.98 µg mt^–1 at 30 min, 0.91 gmt1 at 4 h and 0.59 µg ml^–1 at 9 h, colostrum .plasmaratios at 4 h and 9 h being 0.67 and 0.68, respectively. Meanconcentrations of etomidate in colostrum were 79.2 ng ml^–1at 30 min and 16.3 ng ml^–1 at 2 h. being undetectableat 4 h. The colostrum:plasma etomidate concentration ratio was1.2 at 30 min. We conclude that, although plasma and colostrumconcentrations of thiopentone and etomidate declined rapidly,the decrease was faster with etomidate.     

RESPIRATORY AND HYPNOTIC EFFECTS OF NITRAZEPAM, DIAZEPAM AND PENTOBARBITONE AND THEIR SOLVENTS IN THE RABBIT AND THE MOUSE

Nitrazepam and diazepam only cause significant depression ofrespiratory rate and loss or righting reflex, in mice, at lethaldoses. In sub-lethal doses they induce a stage of deep but rousablesleep which is quite distinct from the intermittent dozing ofcontrol mice and the comatose state of mice injected with pentobarbitone.However, the respiratory effects of nitrazepam and diazepamin high doses in mice and low doses in rabbits are modifiedby the presence of solvent. Furthermore the solvent itself causesa loss of righting reflex in mice and the lethal dose of nitrazepamand diazepam is greatly reduced when the drugs are administeredin solvent     

OBSERVATIONS OF BLOOD LACTATE AND PYRUVATE LEVELS AND EXCESS LACTATE PRODUCTION DURING AND AFTER ANAESTHESIA WITH AND WITHOUT HYPERVENTILATION

Arterial blood levels of lactate and pyruvate and the productionof excess lactate were studied in two groups of patients. Onegroup of ten patients was anaesthetized with <1 per centhalothane, nitrous oxide and oxygen and allowed to breathe spontaneously.The second group of twelve patients was anaesthetized with nitrousoxide and oxygen, and additional trichloroethylene or papaveretum,and deliberately hyperventilated. No significant changes werefound in lactate or pyruvate during anaesthesia in the firstgroup, but there was a significant increase in the lactate:pyruvate ratio half an hour after recovery from the anaesthetic.Significant excess lactate production occurred during anaesthesiain the second group after hyperventilation had been in progressfor half an hour. The excess lactate increased after 1 hourof hyperventilation and further increases were found half anhour after recovery from the anaesthetic. These findings arediscussed in relation to the observed changes in blood-gas levelsand acid-base balance and to the current knowledge of carbohydratemetabolism.     

STRESS AND AROUSAL CIRCADIAN RHYTHMS IN SMOKERS AND NON-SMOKERS WORKING DAY AND NIGHT SHIFTS

Twenty smokers and twenty non-smokers completed a brief Mood State Questionnaire every 2 hours, over one day shift and one night shift. The subjects comprised male police officers and factory workers, aged 23–57 years. Cigarette smokers reported significantly higher stress levels than non-smokers on both day and night shifts (p<0.05). This confirms previous findings that smokers are often more stressed than non-smokers. Stress levels varied over time within each shift (p<0.001), but the circadian patterns did not differ between smokers and non-smokers. Thus smoking did not alter circadian mood rhythms, nor did it facilitate stress control. Self-rated levels of arousal showed the archetypal inverted-U pattern over time in both smokers and non-smokers. There was no difference in mean arousal levels between subgroups, indicating that cigarettes did not lead to greater alertness. The shift×time interaction was significant for both stress (p<0.001) and arousal (p<0.003), indicating different circadian rhythms during the day shift and the night shift. Finally, while smokers consumed slightly more cigarettes during the night shift than day shift (22.3, 19.4 respectively, p<0.05), mean stress and arousal levels did not differ between shifts. The implications of these findings for smoking behaviour are discussed. © 1997 by John Wiley & Sons, Ltd.     

多发、复杂肝内胆管结石的诊治

目的探讨多发性、复杂性肝内胆管结石的诊治方法。方法对我院近8年来的23例多发复杂性肝内胆管结石的病人诊治经验结合文献进行分析。结果这些病例手术后残留结石率高达65．2％,比一般的肝内胆管结石术后残留率30．36％高得多,且严重并发症发生并死亡率高达17．4％。结论多发、复杂性的肝内胆管结石病人的诊治是长期来难以突破的重大临床问题,术前的“个体化”诊疗方案和术式选择是提高疗效的关键。     

VENTILATION AND GAS EXCHANGE DURING ANAESTHESIA AND SURGERY IN SPONTANEOUSLY BREATHING INFANTS AND CHILDREN

Minute ventilation (VE) (mlmin^–1), respiratory frequency(f), mixed expired carbon dioxide fraction (FCO₂ and end-tidalcarbon dioxide concentration E'CO₂) (%) were measured, and alveolarventilation (VA), deadspace (VD), deadspace/tidal volume ratio(VD/VT) and carbon dioxide output (VCO₂) calculated in 58 anaesthetized,spontaneously breathing infants and children weighing 2.8–20.5kg.Although minute volumes varied, tidal volume correlated wellwith weight (r = 0.83), with a mean tidal volume (± ISD)of 5.2±1.2mlkg^–1. It was concluded that, by theuse of mean VT + ISD (approximated to 6 ml kg^–1) the freshgas flow in mlmin^–1 should be set at 2.5x6xkgxf(15xkgxf)to avoid rebreathing in various T-piece systems in anaesthetized,intubated and spontaneously breathing infants up to a body weightof 20 kg. End-tidal carbon dioxide concentration was lower inyounger patients who were premedicated with atropine alone thanin the older ones who received opioid premedication also. Respiratoryfrequency, VD/VT and total VD per minute were higher in theyounger age group, which explained the finding of a high VEin relation to VCO₂ for these patients. This inefficiency ofventilation emphasizes the need to minimize apparatus deadspacein breathing systems used for small infants. ^*Department of Anaesthesia, University Hospital, S-22185 Lund,Sweden. Department of Anaesthesia, St George's Hospital, Blackshaw Road,London SW17.