紫菀药材的高效液相色谱指纹图谱与定量分析

目的:建立紫菀药材的反相液相色谱定性和定量鉴别分析方法.方法:Ultrasphere ODS C18色谱柱(250mm ×4.6mm,5μm),流动相为甲醇-乙腈(1:2)-0.4%磷酸梯度洗脱,流速为1.0 mL/min,检测波长为360 nm,柱温为25℃.紫菀药材经80%甲醇超声处理,进行指纹图谱分析,并对紫菀药材中的主要成分槲皮素进行含量测定.结果:在该色谱条件下,6份不同紫菀药材的RP-HPLC指纹图谱中可检出10个共有峰作为定性鉴别;紫菀药材中槲皮素的含量在0.135%～0.246%之间.结论:本方法样品处理简便,准确性好,可用于紫菀药材的质量控制.     

N2肺癌预后因素分析及手术适应证的探讨

目的 :识别纵隔淋巴结转移 (N2 )的非小细胞肺癌 (non smallcelllungcancer ,NSCLC)患者根治术后的预后不利因素 ,探讨N2 肺癌合理的手术适应证。方法 :回顾1993年 1月 1日～ 2 0 0 0年 1月 3 1日 ,2 12例根治性切除及系统性淋巴结廓清术后病理诊断为N2 肺癌患者的临床资料。应用Ka plan Meier法计算生存率 ,Log rank检验比较不同临床病理因素组间患者的生存差别 ;Cox多因素回归分析 ,判定N2 肺癌预后的不利因素。结果 :本研究 5年生存率为17% ,其中有N2 临床症状、大细胞肺癌和T4患者的预后最差 ,中位生存期不足 12个月。有纵隔淋巴结转移症状 (RR =9 2 ,P <0 0 0 1)、T4肿瘤 (RR =7 15 ,P <0 0 0 1)、淋巴结被膜受侵 (RR =1 72 8,P =0 0 0 8)和多站(≥ 3站 )淋巴结转移 (RR =2 2 94,P =0 0 0 2 )是预后的不利因素。结论 :N2 非小细胞肺癌患者的预后有很大的差别 ,根治切除的适应证应该加以选择。有明显纵隔淋巴结转移症状、大细胞肺癌和T4肿瘤的N2 患者预后极差 ,不宜将外科手术作为首选的治疗方法。     

63例宫颈腺癌临床治疗和预后分析

目的:了解影响宫颈腺癌预后的相关因素,探讨宫颈腺癌比较理想的治疗方法.方法:对天津医科大学附属肿瘤医院1980年1月至2000年1月间收治的63例宫颈腺癌进行分析,占同期宫颈癌的4.65%,并随机选取同期治疗的80例宫颈鳞癌作为对照进行比较.患者平均发病年龄53.7岁,绝经后患者占55.6%.主要症状为阴道不规则出血和/或白带增多.其中Ⅰ期17例,Ⅱ期33例,Ⅲ期13例.20例采用单纯放射治疗,43例采用放射治疗与手术相结合的综合治疗.结果:收治的宫颈腺癌占宫颈癌的比例从80年代的3.5%(38/1087)上升至90年代的7.36%(34/462),其5年总的生存率为56.9%(33/63-5),低于同期宫颈鳞癌5年生存率.单纯放疗组5年生存率50%;手术放疗组5年生存率为60.5%,两组间无统计学差异.行根治性手术的Ⅰ、Ⅱ期患者5年生存率为80%,而行全宫切除的同期患者5年生存率仅为39.3%,但二者差异尚无统计学意义.宫颈肿瘤≥4cm和＜4cm者5年生存率分别为34.3%(12/35)和60.7%(17/28)具有明显差异(P＜0.05).Ⅰ、Ⅱ期5年生存率62%(31/50),而Ⅲ期5年生存率为7.7%(1/13),二者差异显著(P＜0.01).结论:宫颈腺癌的发病比例呈上升趋势,宫颈腺癌的预后与临床分期、肿瘤大小、治疗方式关系密切.以根治性手术为主的综合治疗是宫颈腺癌的主要治疗方法.     

贲门癌切除机械吻合术后的胃食管反流

目的:探讨贲门癌切除食管胃机械吻合术后重建食管与胃食管反流之间的关系.方法:对30例术后患者进行食管胃压力测定,其中16例行24h食管pH监测,12例行内镜检查和病理学检查.结果:测压结果显示:吻合口与吻合口下方的静息压相似,比较差异无显著性(P＞0.05),而吻合口上方静息压增高,平均为3.42mmHg,经比较吻合口上方和吻合口下方差异有显著性(P＜0.05).值得注意的是其压力值(3.42mmHg)远低于正常括约肌的静息压(10～45mmHg),因此其抗反流作用的程度是有限的.本组16例术后食管24h pH监测表明食管的酸暴露时间延长,食管酸暴露时间百分比平均为13.78%,为正常人的11倍,反流次数增加,最长反流时间达43min,说明手术后的胃食管反流是客观存在的.内镜检查和病理检查,83.3%有异常现象及食管炎征象,进一步证实术后患者有不同程度的反流性食管炎.结论:1)贲门癌切除食管胃吻合术后存在胃食管反流.2)反流的发生不因机械吻合或手工吻合而异.3)反流的发生与术后时间长短无关.4)24h食管pH监测是最敏感的观察方法.5)半卧位睡眠是预防反流的有效方法.6)反流的治疗主要应用促动力药和粘膜保护剂.     

Interleukin-12 enhances the sensitivity of human osteosarcoma cells to 4-hydroperoxycyclophosphamide by a mechanism involving the Fas/Fas-ligand pathway.

Cyclophosphamide (CY) and its derivative ifosfamide are alkylating agents used to treat osteosarcoma (OS). The purpose of these studies was to determine whether alkylating agents affect the expression of Fas ligand (FasL) and whether interleukin 12 enhances the sensitivity of human OS cells to alkylating agents. 4-Hydroperoxycyclophosphamide (4-HC), the preactivated CY compound, and 4-hydroperoxydidechlorocloclophosphamide (4-HDC), its nonalkylating analogue, human OS LM6 cells, and a clone of cells derived by transfection with the interleukin 12 gene (LM6-#6) were used for these studies. Incubation of LM6 and LM6-#6 with 10 micro M 4-HC increased the expression of FasL mRNA (2.5- and 3.0-fold, respectively). By contrast, 4-HDC, Adriamycin (ADR), cisplatin (CDP), and methotrexate (MTX) had no effect on FasL mRNA expression. Increased FasL expression after treatment with 4-HC was also demonstrated by immunohistochemistry and flow cytometry. Drug-induced FasL was functional and mediated cell death. We examined the effect of FasL up-regulation by 4-HC on LM6 and LM6-#6 cells. Flow cytometry showed that LM6-#6 cells expressed 2.2-fold more Fas than LM6 cells. Cytotoxicity of 4-HC, 4-HDC, ADR, CDP, and MTX on LM6, LM6-neo, and LM6-#6 were quantified. Colony-forming assay revealed an IC(50) of 2.10 micro M for 4-HC in LM6-neo cells compared with 0.41 micro M in LM6-#6 cells. The IC(50) for 4-HDC, ADR, CDP, and MTX were not significantly different between the two cell lines. We concluded that the increased expression of Fas enhanced LM6-#6 sensitivity to 4-HC. These data indicate that Fas/FasL may be involved in the cytotoxic pathway of CY. Combining biological agents with chemotherapeutic agents that have complementary Fas/FasL pathway actions may offer new therapeutic alternatives.     

Overexpressed eIF4E is functionally active in surgical margins of head and neck cancer patients via activation of the Akt/mammalian target of rapamycin pathway.

PURPOSE: Overexpression of eIF4E in surgical margins of head and neck cancer patients is an independent risk factor for recurrence. We hypothesize that overexpressed eIF4E is functionally active in tumor margins through activation of the Akt/mammalian target of rapamycin (mTOR) pathway EXPERIMENTAL DESIGN: Western blots and/or immunohistochemistry were performed to determine whether phosphorylation of mTOR and activation of its downstream molecules eIF4E-binding protein-1 (4E-BP1) and p70 S6 kinase and the upstream modulator of mTOR, Akt, were expressed in margins overexpressing eIF4E. RESULTS: There was a significant association between phospho-4E-BP1 and eIF4E expression of a margin or a significant difference in phospho-4E-BP1 expression between the eIF4E-positive and -negative margins (P < 0.01). A significant association between eIF4E and phospho-p70 S6 kinase as well as eIF4E and phospho-mTOR was also noted (P < 0.05). Western blot analysis indicated a highly significant difference in the phosphorylation status of 4E-BP1 between tumors and resection margins. A total of 89% of the 4E-BP1-expressing margins expressed more of the phosphorylated (beta, gamma, and delta) isoforms, whereas 81% of the 4E-BP1-expressing tumors expressed more of the unphosphorylated alpha isoform. A similar difference in Akt activation was noted between eIF4E-positive margins and tumors (P < 0.05). CONCLUSIONS: Overexpression of eIF4E is functionally active in tumor margins through activation of the Akt/mTOR signaling pathway. The greater degree of expression of downstream targets and upstream regulators of mTOR in margins compared with the tumors indicates preferential activation of the Akt/mTOR signaling pathway in margins overexpressing eIF4E. Rapamycin analogs can potentially be used as adjuvant therapy for patients with eIF4E-positive margins.     

Porcine stem cell engraftment and seeding of murine thymus with class II+ cells in mice expressing porcine cytokines: toward tolerance induction across discordant xenogeneic barriers

BACKGROUND: Mixed hematopoietic chimerism is a reliable means of tolerance induction, but its utility has not been demonstrated in discordant xenogeneic combinations because of the difficulty in achieving lasting hematopoietic engraftment. Miniature swine are likely to be suitable organ donors for humans. To evaluate the ability of mixed chimerism to induce swine-specific tolerance in widely disparate xenogeneic recipients, this study aimed to achieve long-lasting chimerism in a pig to mouse combination. METHODS: Immunodeficient transgenic mice were developed by crossing transgenic founders carrying porcine interleukin-3, granulocyte macrophage-colony stimulating factor, and stem cell factor genes with severe combined immunodeficient mice or non-obese diabetic/severe combined immunodeficient mice. Swine bone marrow transplantation was performed in these mice, and porcine chimerism was followed for 20 weeks. RESULTS: Whereas swine cells became undetectable in all non-Tg littermates by 7 weeks, high levels of porcine hematopoietic chimerism, including the presence of porcine class II+ cells in the host thymus were maintained in Tg mice for >20 weeks. Colony-forming assays revealed the presence of large numbers of swine hematopoietic progenitor cells in the marrow of these mice at 20 weeks after bone marrow transplantation. CONCLUSIONS: These transgenic mice demonstrate for the first time that spontaneous migration of marrow donor antigen-presenting cells to an intact recipient thymus can occur and that porcine stem cells can persist in this highly disparate species combination. These data therefore support the feasibility of the eventual goal of tolerance induction by mixed chimerism in discordant xenogeneic combinations.     

Morphologic changes in efferent ductules and epididymis in estrogen receptor-alpha knockout mice

Estrogen has been shown to have an important role in fluid reabsorption in efferent ductules of the testis. Our previous study of the estrogen receptor-alpha knockout mouse (ERKO) showed that the efferent ductules and rete testis were primary targets of estrogen receptor function. In the present study, a more comprehensive evaluation of the ERKO male reproductive tract was performed to determine the severity of effects in efferent ductules as well as the epididymis. The following observations were found in ERKO males: 1) blind-ending efferent ductules were more prevalent in ERKO than in wild type (WT) tissues; 2) glycogen-containing cells were observed at the rete testis-efferent ductule junction; 3) the tubular diameters of the efferent ductules and initial segment epididymides were dilated; 4) efferent ductules were dilated between 130 to 300% over wild type ductules; 5) efferent ductule epithelial height was reduced nearly 50%; 6) microvilli of nonciliated cells of efferent ductules were 64% shorter in length; 7) cilia were reduced in number; 8) initial segment epithelium was displaced into regions adjacent to the rete testis and in short segments of the common region of efferent ductule; 9) apical, narrow, and clear cells of the epididymis also were abnormal in some regions; 10) in the corpus and cauda regions, sperm granulomas were noted in one third of the ERKO males. In conclusion, the entire reproductive tract is affected in ERKO males. The cells showing the greatest effects were estrogen receptor-positive cells. It appears that in the ERKO mouse there are developmental anomalies that must be considered separately from adult dysfunctional changes in the male reproductive tract.     

The treatment of premature arrest of growth plate with a novel engineered growth plate: experimental studies

OBJECTIVES: To observe the role of the engineered growth plate (EGP) in the treatment of premature arrest of growth plate and to establish a novel treatment method for the premature arrest of the growth plate. METHODS: The engineered growth plates were cultured for the first time by using polylactic acid (PLA) as cell scaffold seeded with growth plate chondrocytes and they were implanted into the medial proximal defects of growth plates of New Zealand rabbit tibia. The degree of deformity of the tibia was evaluated by X-ray and the expression of collagen II mRNA of regenerating growth plate was detected by in situ hybridization technique. RESULTS: Little deformity appeared in the EGP group 8 weeks after operation. Some deformity was seen in the EGP group 16 weeks after operation, whereas it's degree was much less than that of the control group (P = 0.0001). The degree of the angular deformity of the EGP precultured with bFGF and TGF-beta group was less than that of the EGP group (P < 0.05). The cells in the regenerating growth plate arranged in column and were stained blue by in situ hybridization technique. CONCLUSION: The EGP can prevent the formation of bony bridge and restore the growth of the damaged growth plate.     

糖皮质激素诱导性股骨头坏死模型的血管改变

目的　探讨在糖皮质激素诱导股骨头坏死过程中血管形态的改变与髓腔内脂肪填塞的关系。　方法　 12～ 18个月龄的健康日本大耳白兔 5 2只 ,按 2 5mg·kg-1·d-1注射地塞米松建立股骨头坏死模型 ,通过血管铸型扫描电镜观察、墨汁灌注切片光镜观察及图像分析等方法观察用药中和停药后股骨头病理和血管形态的变化。　结果　用药 2周出现股骨头髓腔内脂肪填塞 ,窦状隙血管因脂肪细胞压迫变细 ,血管铸型、墨汁灌注切片显示了血管表面压迹。病变随用药时间的延长而加重 ,窦状隙血管逐渐失去其结构特征。图像分析示股骨头内血管面积进行性下降 ,8周约为对照组的1/ 4。停药 6周负重区髓腔内仍存在明显脂肪化 ,血管纤细、稀少。实验 4周 ,股骨头内出现典型骨坏死灶。随时间的延长 ,坏死灶增大、股骨头坏死程度加重。　结论　髓腔内脂肪填塞是导致糖皮质激素诱导性股骨头坏死中缺血损害的重要病理因素之一 ,尤其在股骨头坏死的早期起着非常重要的作用