L-arginine对局灶脑缺血脑细胞凋亡和微血管密度的影响

目的探讨NO合成底物左旋-精氨酸(L-Arg)对兔局灶脑缺血后血管再生和脑细胞凋亡的影响。方法兔局灶脑缺血后应用L-Arg,流式细胞仪定量分析细胞凋亡率的变化,CD34免疫组织化学测脑组织微血管密度(MVD),脑组织含水率评价脑水肿。结果与对照组比较,L-Arg组脑细胞凋亡率明显减少(8.72±2.62 vs 16.62±2.82,P<0.01),同时脑组织MVD却明显增加(1.21±0.43 vs 0.69±0.22,P<0.01)。结论外源性L-Arg可减少缺血后脑细胞凋亡并促进缺血后血管再生,对局灶脑缺血具有重要的神经保护作用。     

Segmental colonic transit studies: comparison of a radiological and a scintigraphic method

OBJECTIVE: Colonic transit studies are used to diagnose slow transit constipation (STC) and to evaluate segmental colonic transit before segmental or subtotal colectomy. The aim of the study was to compare a single X-ray radio-opaque marker method with a scintigraphic technique to assess total and segmental colonic transit in patients with STC. METHOD: Thirty-one female patients (median age 46 years) with severe constipation and a prolonged or borderline prolonged colonic transit time on radio-opaque marker study were included in the study. They were subsequently investigated with (111)Indium-DTPA colonic transit scintigraphy, with a median time between the investigations of 4(range 1-27) months. Normal values of healthy female controls were used for comparison. RESULTS: There was no difference between the two methods in terms of prolonged or normal total colonic transit time. Twenty-nine of 31 female patients had a prolonged transit time only in one or two segments on the marker study. On scintigraphy, the transit time was prolonged for patients in the left (P < 0.05 to P < 0.001), but not in the right colon. With respect to prolonged or normal segmental transit time, there was a significant difference between the two methods only in the descending colon (P = 0.02). However, the results varied considerably for individual patients. CONCLUSION: Segmental colonic delay was a common finding. The two methods gave similar results for groups of patients, except in the descending colon. The variation of the results for individuals suggests that a repeated transit test may improve the assessment of total and segmental transit.     

前列腺癌中PIM-1的表达及其临床意义

目的 探讨PIM-1在前列腺癌中的表达及临床意义。方法 逆转录-聚合酶链反应（RT—PCR）半定量分析2例良性前列腺增生（BPH）和5例前列腺癌（PCa）组织标本中PIM-1mRNA表达，免疫组织化学法检测20例BPH、20例高分级前列腺上皮内瘤（HGPIN）和42例PCa组织标本中PIM-1蛋白表达水平，染色结果分为阴性、弱阳性、阳性和强阳性。结果 5例PCa组织PIM-1mRNA表达相对值分别为0．63、0．55、0．42、0．91、0．76，2例BPH中其相对值为0．26、0．27。BPH、HGPIN和PCa组织中PIM-1蛋白阴性表达率分别为60％（12／20）、20％（4／20）和2％（1／42），弱阳性表达率分别为40％（8／12）、20％（4／20）和12％（5／42），阳性列强阳性表达率分别为0（0／20）、60％（12／20）和86％（36／42），PCa中PIM-1蛋白表达水平高于HGPIN和BPH（P值均〈0．05）。PIM-1蛋白表达水平随PCa的临床分期和病理分级增高而增强，在有和没有淋巴结转移PCa组织中PIM-1强阳性表达率分别为70％（7／10）、25％（8／32），差异有统计学意义（P〈0．05）。结论PIM-1高表达可能与PCa发生和发展相关，PIM-1表达水平与PCa分期、Gleason评分呈正相关，可能成为PCa预后判断的肿瘤标志物。     

痔组织弹性纤维退变和血管生成的机制及其意义

目的研究痔组织弹性纤维的病理变化及微血管密度(MVD)和血管生成相关蛋白的表达与痔形成的关系。方法利用免疫组织化学染色技术(SP法),对比研究24例Ⅲ度痔患者的正常肛垫和痔两种组织内弹性纤维和MVD的变化,并检测一氧化氮合酶(NOS)的3种亚型、血管内皮生长因子(VEGF)和金属基质蛋白酶(MMP)2、MMP9在两种组织中表达的差异。结果痔组织存在显著的新生血管形成,痔组织与相对正常肛垫组织间的MVD计数、VEGF、MMP9表达差异存在统计学意义(P<0.05)。痔组织中iNOS明显增高,但与正常肛垫组织比较,不具备统计学意义;痔组织弹性纤维出现断裂、扭曲、变形、玻璃样变等异常,而正常肛垫组织中弹性纤维形态较规则、密集,断裂和变形少见。结论新生血管形成可能是痔的发病机制之一;MMP9对肛垫纤维支持结构的直接降解作用可能是痔形成和加重的一个重要机制;痔组织中iNOS表达增高还提示炎症因素和一氧化氮可能参与了痔的病理过程。     

The value of MCA, CA 15-3, CEA and CA-125 for discrimination between metastatic breast cancer and adenocarcinoma of other primary sites

MCA, CA 15-3, CEA and CA 125 were determined in the serum of 49 patients with metastatic breast cancer and 38 patients with metastatic adenocarcinoma of other primary sites. By using the 99th percentile of the normal value distribution as the cut-off point, the positive predictive value (PV+) was found to be 85% (95% CI 76-94) for MCA, and 71% (95% CI 61-81) for CA 15-3. When receiver-operating-characteristic (ROC) curves were constructed, the PV+ for CA 15-3 was increased to 82% (95% CI 72-92), using 60 U ml-1 as the cut-off point. With the exception of two patients who had a slightly elevated MCA, MCA and CA 15-3 identified the same patients with breast cancer. By combining a positive MCA or CA 15-3 with a negative CEA and CA 125, further improvement of the PV+ could be achieved; 100% (95% CI 91-100). We conclude that MCA and CA 15-3 may play a useful role in discrimination between patients with metastatic breast cancer and those with adenocarcinoma of other primary sites.     

BILIARY HYPERPRESSURE IN RAT EXTRAHEPATIC CHOLESTASIS ALTERS HORSERADISH PEROXIDASE BILIARY EXCRETION

1. The authors investigated the effect of two extrahepatic cholestasis models (one by bile duct ligation and the other by choledocho-jugular fistula) on the hepatic clearance of horseradish peroxidase in male Sprague-Dawley rats divided into four groups. 2. In groups A (n = 5 rats) and B (n = 5), bile duct ligation was performed, while a choledocho-jugular fistula was created in groups C (n = 5) and D (n= 7). A 10 mg intravenous bolus of horseradish peroxidase was injected after 24 h (groups A and C), 48 h (groups B and D) or 1 h (Group E; five sham-operated rats). Serum and bile samples were then serially collected for 2 h. 3. In all groups, serum horseradish peroxidase levels increased soon after injection and then rapidly decreased, the curves being similar. Biliary excretion increased for 30 min and then slowly decreased. The highest horseradish peroxidase biliary concentrations and outputs were found in Group B followed by Group A; both groups had significantly higher levels than Group E. No difference was found between horseradish peroxidase biliary excretion of groups C and D and that of sham-operated rats. 4. When each group was considered separately, sampling times correlated with the corresponding ratios of bile/ plasma HRP. Significant differences were found between the relative slopes of groups A, B and E, but not between those of groups C, D and E. 5. In conclusion, bile duct obstruction greatly affects the plasma-bile transfer of fluid phase markers, such as horseradish peroxidase, while single retention, caused by choledocho-jugular fistula, has no influence. The increased biliary hyperpressure related to the duration of cholestasis may account for the degree of horseradish peroxidase transfer which, in turn, probably depends on an enhanced paracellular passage.     

微血管密度与胃癌侵袭和转移的关系

目的 :探讨胃癌组织中微血管密度 (MVD)与胃癌生物学行为的关系。方法 :应用免疫组织化学S -P法和抗人因子Ⅷ相关抗原 (FⅧ -RAg)的抗体 ,标记 6 8例胃癌组织中的微血管 ,检测MVD。分析其与胃癌组织学类型、分化水平、浸润深度及淋巴结转移的关系。结果 :6 8例胃癌组织中FⅧ -RAg阳性表达 5 2例 ,占 76 .4 7%。FⅧ -RAg和MVD与分化水平、浸润深度 (P <0 .0 1)及淋巴结转移 (P <0 .0 0 1)密切相关。不同组织学类型中FⅧ-RAg的表达具有显著相关性 (P <0 .0 1) ,而MVD则与胃癌组织学类型无显著差异 (P >0 .0 5 )。 结论 :血管生成对胃癌的发展具有一定作用 ,FⅧ -RAg的检测和MVD可作为判断胃癌恶性程度和预后的生物学指标。     

不同浓度5-溴脱氧尿嘧啶核苷标记山羊骨髓基质干细胞的体外研究

Objective To explore the feasibility of labeling and tracing in vitro goat bone marrow mesenchymal stem cells (BMSCs) by bromodeoxyuridine (BrdU) on the basis of investigation of its optimal concentration, incubating time and cytotoxicity. Methods A healthy goat, aged 10 months old, male, weighing 32 kg, was used in this study. Bone marrow was aspirated. BMSCs were isolated and cultured using the adherence method in vitro. The fourth passage of BMSCs (P4) were incubated with BrdU at 5, 10, 15, 20 μmol/L as 5, 10, 15, 20 μmol/L BMSC groups. Cells were not labeled by BrdU as negative control. The following parameters were measured: induction, differentiation and determination of goat BMSCs; the optimal mass concentration and incubation time of 5-BrdU labeling; cell positive rate at 12, 24, 48 and 72 hours in each group using immunofluoreseenee; the cell survival rate after various concentrations of BrdU ladling by trypan-blue exclusion. Results The morphology of the primary and passage goat BMSCs was fusiform in shape. Goat BMSCs could differentiate into osteoblasts and chondrocytes following induction. BMSC nucleus showed green fluorescence under fluorescence microscope after being labeled by BrdU. The mean labeling rate increased with the increase in the concentration and incubation time of BrdU, and reached to (93.32± 3.25)% after incubation in 15 μmol/L, BrdU for 48 hours. There were no significant differences between 15 μmol/L BrdU for 72 hours, 20 μmol/L BrdU for 48 hours and 72 hours (P > 0.05), or between the other groups or time points (P < 0.05). The labeling rate of the blank control group was 0. The cell survival rate was all above 90% (P > 0.05). Conclusions BrdU can be used as a labeling marker for goat BMSCs. When the concentration is 15 μmol/L and the incubation time is 48 hours, the optimal labeling effect can be achieved. Goat BMSCs labeled with BrdU is of high efficiency and safety.     

RGD-based strategies for selective delivery of therapeutics and imaging agents to the tumour vasculature

During the past decade, RGD-peptides have become a popular tool for the targeting of drugs and imaging agents to alphavbeta3-integrin expressing tumour vasculature. RGD-peptides have been introduced by recombinant means into therapeutic proteins and viruses. Chemical means have been applied to couple RGD-peptides and RGD-mimetics to liposomes, polymers, peptides, small molecule drugs and radiotracers. Some of these products show impressive results in preclinical animal models and a RGD targeted radiotracer has already successfully been tested in humans for the visualization of alphavbeta3-integrin, which demonstrates the feasibility of this approach. This review will summarize the structural requirements for RGD-peptides and RGD-mimetics as ligands for alphavbeta3. We will show how they have been introduced in the various types of constructs by chemical and recombinant techniques. The importance of multivalent RGD-constructs for high affinity binding and internalization will be highlighted. Furthermore the in vitro and in vivo efficacy of RGD-targeted therapeutics and diagnostics reported in recent years will be reviewed.