DNA探针的高效标记和哺乳动物细胞转化子的检测

本实验采用随机引物法标记探针DNA.探针DNA加热变性成单链,以单链DNA为模板,六聚脱氧核苷酸为引物,在DNA多聚酶I大片段的作用下进行放射性标记.标记探针的比放射性达10~8cpm/μg DNA以上.放射性底物的掺入率为60%.用该方法标记的人高度重复顺序探针,可以检出微微克水平的阳性分子.与人HeLa S_3细胞DNA转化的小鼠LTK~+细胞DNA打点杂交呈阳性,与未经转化的小鼠LTK~-细胞DNA杂交为阴性,证明小鼠转化细胞中存在HeLa细胞DNA,DNA转化是成功的.     

Cerebral hemodynamic and metabolic profiles in fulminant hepatic failure: relationship to outcome.

The purpose of this retrospective study was to examine the potential role of cerebral hemodynamic and metabolic factors in the outcome of patients with fulminant hepatic failure (FHF). Based on the literature, a hypothetical model was proposed in which physiologic changes progress sequentially in five phases, as defined by intracranial pressure (ICP) and cerebral blood flow (CBF) measurements. Seventy-six cerebral physiologic profiles were obtained in 26 patients (2 to 5 studies each) within 6 days of FHF diagnosis. ICP was continuously measured by an extradural fiber optic monitor. Global CBF estimates were obtained by xenon clearance techniques. Jugular venous and peripheral artery catheters permitted calculation of cerebral arteriovenous oxygen differences (AVDO2), from which cerebral metabolic rate for oxygen (CMRO2) was derived. A depressed CMRO2 was found in all patients. There was no evidence of cerebral ischemia as indicated by elevated AVDO2s. Instead, over 65% of the patients revealed cerebral hyperemia. Eight of the 26 patients underwent orthotopic liver transplantation-all recovered neurologically, including 6 with elevated ICPs. Of the 18 patients receiving medical treatment only, all 7 with increased ICP died in contrast to 9 survivors whose ICP remained normal (P < 0.004). Hyperemia, per se, was not related to outcome, although it occurred more frequently at the time of ICP elevations. Six patients were studied during brain death. All 6 revealed malignant intracranial hypertension, preceded by hyperemia. In conclusion, the above findings are consistent with the hypothetical model proposed. Prospective longitudinal studies are recommended to determine the precise evolution of the pathophysiologic changes.     

MTT法和水溶性四氮唑(WST-1)法检测中波紫外线照射HaCaT细胞后活力的比较

目的:观察中波紫外线照射后HaCaT细胞活力的变化并探讨水溶性四氮唑(WST-1)法检测的适用性.方法:将HaCaT细胞分为对照组(假照射组)、300J/m2、600J/m2、900J/m2紫外线照射后24h组(每组6个样本).600J/m2紫外线照射后4h组、8h组、1 2h组、24h组、48h组、72h组(每组6个样本).血球计数板计算HaCaT细胞数量.四甲基偶氮唑盐(MTT)法和水溶性四氮唑(WST-1)法活性测定HaCaT细胞活性.结果:MTT法显示300J/m2、600J/m2、900J/m2紫外线照射HaCaT细胞24h后光密度(OD)值明显较0J/m2组低;WST-1法各组OD值明显比0J/m2组低.MTT法显示600J/m2紫外线照射HaCaT细胞8h、12h、24h、48h、72h后OD值明显比0J/m2组低.WST-1法显示600J/m2紫外线照射HaCaT细胞4h后OD值较0J/m2组低;600J/m2紫外线照射HaCaT细胞8h、12h、24h、48h、72h后OD值明显比0J/m2组低.WST-1法的OD值与细胞计数、MT法OD值呈正相关.结论:随着紫外线剂量的加大和照射后时间延长,HaCaT细胞数量减少,细胞的活力下降,WST-1代谢活性亦降低,呈时间和剂量依赖性.HaCaT细胞可对WST-1进行代谢,适用于该细胞增殖研究.     

Saccular Aneurysm of the Azygos Anterior Cerebral Artery: Three Case Reports

The azygos anterior cerebral artery, a rare anomaly in the circle of Willis in which only a single vessel supplies the medial aspects of both anterior cerebral hemispheres, is closely associated with saccular aneurysms. We present three cases of azygos anterior cerebral artery aneurysms among the 781 cerebral aneurysms surgically treated at our institution in an 11-year period. These three cases all involved elderly women who presented with subarachnoid hemorrhage. Conventional cerebral angiography and CT angiography revealed small saccular aneurysms at the distal ends of the azygos anterior cerebral arteries. These aneurysms were clipped successfully using a bifrontal interhemispheric approach. Hence, the pathogenesis of these particular aneurysms relating to hemodynamic change, associated anomalies, and surgical pitfalls is discussed with review of literature.     

The tuberous breast revisited.

BACKGROUND: The tuberous breast presents a problem for which many surgical solutions have been described. Current teaching describes how the tuberous breast deformity is the result of skin shortage as well as herniation of breast tissue through the nipple-areola complex. However, through careful clinical observation we now believe that the only abnormality present is herniation of breast tissue through the nipple-areola complex. METHODS: Using this principle, we have refined a one-stage surgical procedure that can be used to correct any type of tuberous breast deformity. Since 2001 we have performed our technique on a series of 13 tuberous breasts of widely varying appearances in eight patients (age 17-24 years) with a follow up varying between 3 and 56 months. Our new understanding of the tuberous breast deformity has also made it possible to develop an objective, reproducible method for defining the tuberous breast based on the degree of areola herniation. RESULTS: All patients reported high levels of satisfaction with the procedure. Assessment of the results by an independent panel of attending surgeons showed all results to be good/excellent. Moreover, the results have improved with time and no revisions have been needed. Our method of defining the tuberous breast (based on the ratio of areola herniation:areola diameter) enabled us to identify a cut-off to decide (objectively) when a breast was tuberous. This allowed us to anticipate when an areola reduction/tightening procedure would be necessary to avoid a 'double-bubble' deformity. CONCLUSION: We propose a one-stage surgical procedure which is applicable to all degrees of tuberous breast deformity. The results appear to confirm our theory that the only abnormality present in the tuberous breast is herniation of breast tissue through the nipple-areola complex. In patients with small breasts and a tuberous deformity, correction of the herniation changes the tuberous breast into a simple hypoplastic breast. The volume deficit can then be corrected by augmentation (if desired by the patient). In patients with sufficient breast volume, correction of the herniation alone will correct the deformity.     

Treatment of congenital cystic adenomatoid malformation-does resection in the early postnatal period increase surgical risk?

OBJECTIVE: The recent development of fetal ultrasonography has allowed for an increasing number of prenatal diagnoses for congenital cystic adenomatoid malformation (CCAM). However, the appropriate surgical timing of these patients has not been studied as of yet. The aim of this study is to suggest a safe strategy for the treatment of CCAM by identifying the relationship between the timing of surgery and postoperative outcome. METHODS: Between 1987 and 2003, 40 patients (28 males, 12 females) underwent surgical resection for CCAM. The mean age was 38.6+/-9.1 (2 days-13 years) months. CCAM was diagnosed by prenatal ultrasonography in eight patients. Early operations were performed in four out of the eight. Operation was deferred until 2-12 months of age for the remaining four patients. RESULTS: Type I CCAM was found in 20 patients, type II in 20 and no patient exhibited type III. Five patients had associated pectus excavatum anomaly. There were no cases of operative mortality. Seventeen minor postoperative complications developed in 16 patients (40.0%): prolonged chest tube drain in 10, wound infection in 4, and 1 case of pneumonia, empyema and pleural space, respectively. The average hospital stay was 11.8 (6-29) days. During the mean follow-up period of 67.5 months, one patient died of accidental aspiration 7 months after operation during the postoperative recovery course of Ravich operation for pectus excavatum. The remaining patients reported doing well with normal physical activity. All five patients who underwent surgery at the age of under 1 month did not exhibit increased postoperative morbidity. CONCLUSIONS: We concluded that surgery for CCAM could be safely performed in all age groups with satisfactory long-term outcomes. It is suggested that early elective surgical correction can be recommended for a patient whose diagnosis was made in utero.     

食管癌贲门癌术后胸胃排空障碍的临床分析

目的探讨食管癌贲门癌术后胸胃排空障碍的诱因、诊断和治疗。方法回顾性分析1990年1月-2003年12月我院施行1369例食管癌贲门癌切除术的病人临床资料。结果本组病例发生胸胃排空障碍17例,发生率1.24%,其中机械性胸胃排空障碍5例,功能性胸胃排空障碍12例,均发生于术后3～12d,再次手术治疗5例;治愈16例,死亡1例。结论上消化道造影、胃镜是鉴别机械性胃排空障碍和功能性胃排空障碍的重要方法,对食管癌术后机械性胃排空障碍应采取手术治疗,而功能性胃排空障碍采取保守治疗多能好转。