大鼠原代肝细胞的培养及鉴定

目的:探讨体外大鼠肝细胞的分离培养及纯化的方法。方法:采用胶原酶Ⅳ消化组织块法分离新生大鼠肝细胞,对大鼠肝细胞进行纯化、培养,并采用免疫组织化学方法对其进行鉴定。结果:培养24 h,有部分细胞贴壁,48 h有大量细胞贴壁,但含有较多杂细胞。经过4~5次传代后,细胞形态一致,可见许多双核细胞和多核细胞。采用抗大鼠细胞角蛋白(ck-18)的特异抗体进行免疫细胞化学鉴定,经SP染色DAB显色后,阳性着色部位在胞浆呈棕黄色颗粒,阴性对照未见着色。结论:成功地体外培养了大鼠肝细胞并对其进行了鉴定,为进一步的实验研究奠定了基础。     

Cross-Cultural Applicability of FACES IV

We studied cross-cultural Hungarian adaptation and validation of FACES IV for screening adaptive versus nonadaptive family functioning. A protocol was followed including translation, tests of reliability, construct and convergent validity, standardization, testing of family types, and ratios. Two hundred forty-nine couples (498 adults) from various regions of the country participated in the study. Results indicated high item–remainder correlations and Cronbach alphas. Factor analytic data fitted well with the model indicated by the Olson studies. Most salient results were found for Cohesion, which partially overlapped Flexibility, and had high negative correlations with the Disengaged dimension. Rigid and Chaotic as well as Enmeshed were found to be independent factors. Family types of the Hungarian sample explored by cluster analysis were comparable to the U.S. sample.     

自体胸腔积液上清作为培养液对肿瘤浸润淋巴细胞生长的影响

目的：观察自体胸腔积液上清作为培养液对肿瘤浸润淋巴细胞(TIL)生长的影响．方法：恶性胸腔积液中的TIL经贴壁法分离后，分别采用含有细胞因子白细胞介素-2(IL-2)、CD3单克隆抗体(OKT3)及植物血球凝集素(PHA)的自体胸腔积液上清及含10％人AB型血清的RPMIl640进行培养，比较两种培养液对TIL的增殖速度、培养前后免疫表型变化和对自体肿瘤细胞的杀伤活性。结果：培养2周，经统计学比较发现，自体胸腔积液上清作为培养液对TIL的生长速度、免疫表型和对自体肿瘤细胞杀伤活性的影响与应用含10％人AB型血清的RPMIl640培养液培养无差异。结论：自体胸腔积液上清作为培养液培养TIL是可行的。为自体TIL疗法治疗恶性胸腔积液提供理论基础。     

SERUM-FREE CULTURE OF RHESUS MONKEY EMBRYONIC STEM CELLS

Previous studies have shown that the maintenance and proliferation of undifferentiated rhesus monkey embryonic stem (rES) cells requires medium supplemented with fetal bovine serum (FBS). Due to the uncharacterized composition and variation in serum nature, the present study aimed to replace the serum-containing medium with a serum-free medium in the rES cell culture. The results showed that after the initial 48-h culture in the routinely used serum-containing medium, rES cells can grow and proliferate for a prolonged period in the serum-free medium composed of DMEM supplemented with a cocktail of BSA, IGF-1, TGF- &#102, bFGF, aFGF, estradiol, and progesterone. rES cells cultured in the serum-free medium maintained high level of alkaline phosphatase activity and OCT4 level. There was no indication of differentiation as judged by the marker gene expression of all three embryonic germ layers and trophoblast. In addition, serum-free culture would not affect the passage capacity and differentiation potential of rES cells. This work will facilitate the future study of induced differentiation of rES cells and other applications.     

Increasing research capacity and changing the culture of primary care towards reflective inquiring practice: the experience of the West London Research Network (WeLReN)

A number of primary care research networks were set up throughout England in 1998 in order to (1) improve the quality of primary care research (2) increase the research capacity of primary care, and (3) change the culture of primary care towards reflective inquiring practice (NHSE, 2000b). It is not clear how best to operate a network to achieve these diverse aims. This paper describes the first 30 months of a network that adopted a whole system approach in the belief that this would offer the best chance of simultaneously achieving the three aims. A cycle of activity was designed to facilitate the formation of multidisciplinary coalitions of interest for research with complementary 'top down' and 'bottom up' programmes of work co-existing. At least 330 people participated in the generation of research questions of whom one third (33%) were general practitioners, 16% community nurses, 6% practice managers and other primary care practitioners. Over two fifths (43%) were 'key allies' - academics, health authority staff, community workers and project workers. One fifth (110) of all practices (500) in the WeLReN area have collaborated in at least one research project. The ratio of doctor:nurse participation in the 24 research project teams was markedly different in the supported coalitions (2:1) compared to projects devised and led by more experienced researchers (6:1). The evidence suggests that it is possible to operate a primary care research network in a way that develops coalitions of interest from different parts of the health care system as well as both 'top down' and 'bottom up' led projects. It is too early to tell if the approach will be able to achieve its aims in the long-term but the activity data are encouraging. There is a need for more research on the theoretical basis of network operation.     

Designing personalised cancer treatments

The concept of personalised medicine for cancer is not new. It arguably began with the attempts by Salmon and Hamburger to produce a viable cellular chemosensitivity assay in the 1970s, and continues to this day. While clonogenic assays soon fell out of favour due to their high failure rate, other cellular assays fared better and although they have not entered widespread clinical practice, they have proved to be very useful research tools. For instance, the ATP-based chemosensitivity assay was developed in the early 1990s and is highly standardised. It has proved useful for evaluating new drugs and combinations, and in recent years has been used to understand the molecular basis of drug resistance and sensitivity to anti-cancer drugs.     

组织工程血管管状支架的制备与动态培养在血管组织工程中的应用

目的探索体外构建组织工程血管及细胞种植的方法。方法(1)制备管状支架模具,采用模具成形,冷冻干燥等技术,应用胶原和透明质酸复合体外预构管状支架;(2)分动态种植和静态培养两组进行细胞分层种植,每组12个,应用水溶性磺酸四唑(WST-1)测定,扫描电镜观察,5-溴2-脱氧嘧啶核苷(BrdU)细胞标记鉴定比较两种方法的细胞生长情况。结果(1)成功制备了管状支架,在培养液中约3周可保持管腔形态不变形,不塌陷;2.WST-1测定值在培养6、14、20 d分别为:动态组0.842±0.110、1.124±0.102、1.132±0.112,静态组0.386±0.121、0.489±0.245、0.499±0.265,两组比较差异有统计学意义,电镜和BrdU标记表明动态组细胞排列整齐有序。结论应用模具成形,冷冻干燥技术预构管状支架的方法切实可行,动态培养有助于细胞种植,本研究为组织工程血管的构建方法提供了思路。     

两种市售试剂盒检测无症状男性不育症门诊患者支原体感染及药敏状况

目的了解无症状男性不育症门诊患者支原体感染情况,比较两种试剂盒检测结果的判定符合率和药敏结果。方法使用两种市售试剂盒对303例男性不育症门诊患者的尿道棉拭子进行支原体培养、测定和药敏试验。结果两种试剂盒解脲脲原体（UU）培养阳性率分别为14．85％、15．84％,人型支原体（MH）培养阳性率均为2．97％;UU培养阳性和阴性结果判定总符合率达97．03％,MH总符合率高达100％;两种试剂盒敏感率最高的3种药物分别为交沙霉素（JOS）、强力霉素（DOx）、美满霉素（MIN）和阿奇霉素（AZI）、克拉霉素（CLA）、JOS,耐药率最高的药物均为环丙沙星（CIP）。结论无症状男性不育人群中确实有一定水平的支原体感染率或者携带率,市售两种试剂盒的支原体培养和药敏试验性能比较稳定、结果相对可靠。     

Transient urethral obstruction predisposes to ascending pyelonephritis and tubulo-interstitial disease: studies in rats

Chronic tubulo-interstitial disease, an important cause of end-stage renal disease, often results from the combined effects of a disturbed urinary outflow tract and urinary tract infection. Acute unilateral ureteral obstruction in rats rapidly induces foci of medullary necrosis, confined to the region of the papilla and fornices. This injury may provide a nidus for bacterial invasion and may invoke reactive and regenerative changes, ultimately leading to chronic pyelonephritis and tubulo-interstitial nephropathy. To explore this possibility, adult rats underwent renal morphological evaluation 2–7 days following transient 24-h unilateral ureteral obstruction. In some experiments the bladder was inoculated with bacteria (10⁸–10⁹ cfu/ml Escherichia coli in 0.5 ml) after release of ureteral obstruction, with subsequent cultures obtained from the pelvis of both kidneys and from the urinary bladder. Morphologic evaluation of perfusion-fixed kidneys, 2–7 days after the release of 24-h ureteral obstruction disclosed papillary necrosis, urothelial proliferation, marked inner-stripe interstitial expansion, and fibrosis and proximal tubular (S₃) dilatation. The lateral (perihilar region) was predominantly affected, with lesions spreading from the fornices. There was some progression of interstitial fibrosis during the postobstructive time course or following more prolonged ureteral obstruction. By contrast, infection hardly contributed to the tubulointerstitial changes. In rats subjected to infection, cultures were positive in all 15 postobstructive kidneys, as opposed to five contralateral kidneys (P < 0.0001). Viable counts from the postobstructive kidney were also higher than those from the contralateral side (79,000 ± 12,000 vs 2900 ± 1600 cfu/ml, mean ± SEM, P < 0.0001), and were comparable to those obtained from the bladder (77,000 ± 13,000 cfu/ml). We conclude that transient ureteral obstruction predisposes to ascending pyelonephritis and to tubulointerstitial disease. This vulnerability may relate to altered urodynamics and medullary tissue destruction. Received: 28 December 1999 / Accepted: 28 September 2000