C57BL/6小鼠前列腺癌原位细胞移植动物模型的建立

目的 建立一种C57BL/6小鼠前列腺癌原位细胞移植动物模型.方法 C57BL/6小鼠30只,用微量注射器分别将0.5×106个RM-1细胞注射入前列腺左、右背侧叶包膜下.每3 d随机处死5只小鼠,动态观察小鼠前列腺癌局部生长、盆腔淋巴结转移和器官转移;术后15 d处死小鼠后,剩余5只常规饲养直至死亡,观察小鼠的平均荷瘤生存期.结果 C57BL/6小鼠前列腺原位移植RM-1细胞后第3、6、9、12、15天前列腺体积组间差异有统计学意义(P＜0.01);术后第12天开始,出现明显尿潴留、双侧或单侧输尿管扩张、肾脏体积增大、肾盂扩张;组织学观察发现术后前列腺癌细胞逐渐取代正常的前列腺组织,术后第12天组可见肿瘤侵犯周围肌肉,术后第15天组可见肿瘤侵犯精囊和膀胱.术后第3、6、9天未见明显盆腔淋巴结转移,第12、15天盆腔淋巴结转移率均为80%.各组均未发现明显的远处器官转移.各组小鼠成瘤率均为100%.小鼠平均荷瘤生存期为(15.60±0.89)d.结论 虽然未见明确的器官转移情况,但该模型能够较好的模拟人类前列腺癌的发生、发展、局部侵袭及淋巴转移的过程,是一种较理想的前列腺癌动物模型.

Abstract：

Objective To investigate the regularity in establishing the prostate cancer orthotopic transplantation model in C57BL/6 mice. Methods RM-1 cells (0.5×106)were injected into the right and left dorsal lateral prostate capsules of 30 C57BL/6 male mice respectively using micro-syringe. Five mice were sacrificed every three days to observe the local growth, the occurrence and development of prostate cancer, and the remaining five mice were fed to death to calculate the average survival period of tumorbearing mice. Results The volume of prostates was significantly different among the five groups. From the 12th day, significant retention of urine, ureterectasia (bilateral or unilateral ), increased kidney volume and pelviectasis were observed. The rate of tumor formation in the five groups which included the 3rd, 6th,9th, 12th d and 15th day was all 100%. HE staining showed that prostate cancer cells gradually replaced the normal prostate tissue. At the 12th day, muscle tissue around prostate was invaded by prostate cancer.In some samples there was metastasis of seminal vesicles and bladder at the 15th day. The rate of metastasis in pelvic lymph nodes at the 12th or 15th day was 80%. The average survival period of tumor-bearing mice was (15.60±0.89) days. Conclusion The prostate cancer orthotopic transplantation model in C57BL/6 mice can better simulate the process of occurrence, development, local invasion and lymph node metastasis of human prostate cancer and be suitably used as a model of prostate cancer research.     

Effects of CRL 41034 on the adrenergic neuroeffector interaction in the canine saphenous vein

Experiments were designed to determine the effect of CRL 41034, a buflomedil analogue, on the adrenergic responsiveness of canine veins. Rings of saphenous vein (without endothelium) were suspended for isometric tension recording in modified Krebs-Ringer bicarbonate solution at 37 degrees C. CRL 41034 produced a concentration-dependent inhibition of the contractions evoked by the alpha adrenergic agonists norepinephrine, phenylephrine and UK 14304 which was insensitive to the blockade of neuronal uptake by cocaine. CRL 41034 was more potent in inhibiting the concentration-dependent contractions evoked by UK 14304 than those by phenylephrine and the antagonism it caused against the response to UK 14304 fulfilled the criteria for competitivity. CRL 41034, at 10(-5) M significantly depressed, and at 10(-4) M abolished the contractions induced by electrical stimulation of the adrenergic nerves and those evoked by the indirect sympathomimetic amine tyramine. Strips of canine saphenous vein were superfused after incubation with [3H] norepinephrine. During sympathetic nerve activation, CRL 41304 increased the stimulation-evoked overflow of [3H] norepinephrine and 3-methoxy-4-dihydroxyphenylglycol; in the presence of rauwolscine the compound only increased the stimulation-evoked overflow of 3,4-dihydroxyphenylglycol. These experiments suggest that the major vascular effects of CRL 41034 in canine veins are blockade of alpha 2-adrenoceptors on vascular smooth muscle, and inhibition of prejunctional alpha 2-adrenoceptors on adrenergic nerve endings.     

一种小鼠原代肝细胞培养方法

目的寻求一种简易、廉价的原代小鼠肝细胞培养方法。方法采用Ⅳ型胶原酶消化法制备游离肝细胞悬液，以低血清进行肝细胞单层培养。结果通过多次探索、观察，比较成功地进行了原代肝细胞培养。结论此方法适合一般实验室开展肝细胞培养。     

2008年泌尿外科热点问题

随着科学技术的飞速发展,随着世界逐渐进入老龄化社会,泌尿外科疾病发病率日益增高,泌尿外科逐渐发展成为临床医学的一门重要学科.2008年泌尿外科有许多热点问题.     

雄激素剥夺条件下阻断自噬增加LNCaP细胞凋亡

Objective To investigate the change of apoptosis in LNCaP cells after inhibition of autophagic process under androgen removal conditions. Methods The autophagic level was deter-mined by using confocal microscopy and RT-PCR. The DAPI staining was used to indicate the apopto-sis of LNCaP cells after inhibition of autophagic by 3-MA. Also, Z-VAD-FMK was used to extend the apoptosis results. Results ①Androgen deprivation led to increased autophagy in LNCaP cells. LN-CaP cells cultured in complete medium(CM) presented low autophagic process with 1.9 scores. After 24 hours, the punetate GFP-LC3 structures were accumulated in the cells cultured in serum-free medi-um (SF)(2.64 scores). In contrast, the number of punctate GFP-LC3 remained at a very low level (1.85 scores), when cells were incubated with DHT in SFA(serum-free medium+DHT). Statistical analysis showed the significant difference between SF and SFA (P<0.01). Semiquantitative RT-PCR was employed to examine the mRNA expression of LC3. Indeed, cells grown in the medium without serum had a higher LC3 mRNA expression with the highest at 12 hour time point as compared with the cells grown in CM. DHT treatment reduced the level of LC3 mRNA. ②Blockage of autophagy by 3-MA increased the apoptosis of LNCaP cells. LNCaP cells in SF and SFA just presented a basal level of apoptosis, which is (3.19±1.09)% and (3.01±0.33)% , respectively. Under androgen-free con-ditions, inhibition of autophagy by 3-MA could increase apoptosis significantly(10. 90±2.91%). While Z-VAD-FMK, a pan Caspase inhibitor, was able to suppress this apoptotic process to the level of (1.16±0.52)%, which was statistically significant(P<0.01). Conclusions Androgen removal can lead to the increase of autophagy in LNCaP cells. Moreover, inhibition of autophagy promotes the occurrence of apoptosis.     

病理复查并前列腺癌根治术后早期前列腺特异抗原监测的临床意义

目的：结合病理复查结果，监测前列腺癌根治术后早期的前列腺特异抗原(PSA)水平，指导术后的进一步治疗。方法：对25例前列腺癌根治术患者于术后1、2、4周时测定PSA水平，尔后每个月测定一次，随访5～17个月，平均11个月；并对术后标本的手术切缘及包膜浸润程度行病理复查。结果：病理报告T1～T2期17例，15例获根治，PSA水平均降至0．2μg／L以下，随访12个月无复发；2例PSA一直未降至正常，2个月后复升高而加用辅助治疗，其病理观察发现手术切缘有肿瘤残留。T3a或T3b期8例，4例获根治，PSA在3个月内降至0．2μg／L以下，随访9个月无复发，病理特征为肿瘤浸润包膜但未穿透；4例在1个月后因PSA回升分别加用辅助治疗，病理示肿瘤均穿透包膜。结论：T1～T2期患者，因手术切缘残留等原因，肿瘤可能早期复发；T3期患者，特别是那些侵犯包膜但未穿透者，手术可能达到根治，并可以延迟应用雄激素阻断治疗，延长治疗的有效时间。     

INTERACTION OF EXOGENOUS OUABAIN AND CHRONIC MINERALOCORTICOID TREATMENT IN THE KIDNEY OF THE CONSCIOUS SHEEP

1. Ouabain is known to have natriuretic effects only at high doses, and therefore if endogenously produced ouabain has a role in the regulation of sodium excretion, the renal response to ouabain must be increased substantially in certain physiological situations. The aim of this study is to determine whether treatment with the mineralocorticoid, aldosterone, potentiates that natriuretic response to ouabain. 2. Six conscious sheep received renal arterial infusion of either vehicle or aldosterone (3 μg/h). Forty hours after commencement of infusion ouabain was infused into the renal artery at 400 μg/h for 60min. A second infusion of ouabain was administered on the 6th day of aldosterone treatment. 3. In the absence of aldosterone, the effects on sodium excretion produced by ouabain infusion at 400 μg/h into the renal artery were variable and not statistically significant. Ouabain infusion after 40 h of aldosterone treatment increased sodium excretion from 40 ± 14 to 676 ± 69 μmol/min in the second hour following cessation of ouabain infusion (P< 0.001). Ouabain infusion after 6 days of aldosterone treatment increased sodium excretion similarly. Ouabain-stimulated sodium excretion was significantly greater during aldosterone treatment compared to vehicle treatment (P<0.05). In contrast, no enhancement of effect was observed after acute treatment with aldosterone. 4. These results demonstrate potentiation of the natriuretic response to ouabain infusion by chronic mineralocorticoid treatment and suggest a potential role of endogenous digitalislike factor in the physiological control of sodium homeo stasisaldo sterone, endogenous digitalis-like factor, ouabain, sodium excretion.     

高血压对良性前列腺增生细胞增殖与凋亡的影响

目的:探讨高血压对良性前列腺增生(BPH)细胞增殖与凋亡的影响。　方法:分别采用Ki 67免疫组织化 学染色及核酸末端原位标记技术(TUNEL)对40例单纯BPH和40例BPH合并高血压患者前列腺组织的细胞增殖 指数(proliferativeindex,PI)与凋亡指数(apoptoticindex,AI)进行检测。　结果:在单纯BPH以及BPH合并高血 压组,前列腺上皮和间质细胞的PI均大于AI(P<0.05),且PI与前列腺体积呈正相关(P<0.05)。与单纯BPH组 比较,BPH合并高血压组前列腺上皮和间质细胞的PI明显增高(P<0.05)。在BPH合并高血压组,高血压病程的 长短与前列腺上皮和间质细胞的PI呈正相关(P<0.05)。　结论:高血压尤其是长期高血压状态可促进前列腺 上皮和间质细胞的增殖,进而导致前列腺体积增大。     

肾上腺节细胞神经瘤的临床病理学分析

目的 探讨肾上腺节细胞神经瘤的临床及病理学特点。方法 收集经手术病理证实的13例肾上腺节细胞神经瘤患者临床资料进行分析。患者男5例，女8例。平均年龄38岁。肿瘤位于左侧7例，右侧5例，双侧1例。体检时B超偶然发现6例，上腹部不适或间歇性腹痛6例，高血压表现1例。血液生化检查未见特殊异常。影像学检查提示肾上腺实性占位。肿瘤最大直径平均6．7cm。结果 行开放手术治疗12例，腹腔镜手术治疗1例。13例手术顺利，无并发症发生。病理表现：肿瘤主要由成熟的神经轴突、神经束和纤维组织构成，其间散在成熟的节细胞、Schwann细胞。其中3例肿瘤部分区域细胞分化不成熟，有异形性。肿瘤伴囊性变2例，伴钙化1例。随访时间4～116个月，平均43个月，未见肿瘤局部复发和远处转移。结论 肾上腺节细胞神经瘤是少见的良性肿瘤，具有独特的病理特点，手术治疗效果良好。