中华硬蜱再次叮咬宿主后中肠消化细胞的形态测量学研究

本文用图像分析仪对初次和再次叮咬宿主兔后的中华硬蜱中肠消化细胞进行了ＤＮＡ和形态测量学分析。结果表明；蜱血餐后其中肠消化细胞ＤＮＡ增加，约第５天达高峰。初次叮咬组吸血后ＤＮＡ倍体数增加显著，第五天ＤＮＡ平均倍体数达７０６，峰值达１２Ｃ，≥５Ｃ细胞达４４２％。且随叮咬吸血时间延长，中肠增粗，直径增大；肠壁增厚，肠壁横截面面积增大；消化细胞增多，消化细胞截面数增加；消化细胞增大，消化细胞平均截面面积增大；而消化细胞数面密度下降，消化细胞核的体密度和表面积密度下降。这些结果均是对吸入血液进行消化和储存营养物质的正常反应。当中华硬蜱叮咬经初次感染后的宿主兔时，ＤＮＡ倍体数、横切面肠壁面积、消化细胞截面数、消化细胞平均截面面积等增加的幅度不如初次叮咬组，而消化细胞核的体密度和表面积密度下降更显著，提示由于初次叮咬使宿主产生了较强的免疫力而使中肠消化细胞破坏、脱落或抑制增殖而出现吸血量下降，吸血后的上述各种正常反应降低     

中华硬蜱叮咬免疫兔后中肠消化细胞DNA图像分析

本文用图像分析仪对叮咬经不同抗原免疫接种兔后的中华硬蜱中肠消化细胞ＤＮＡ进行测量分析。结果表明：蜱血餐后其中肠消化细胞ＤＮＡ增加，约第５天达高峰。佐剂对照组吸血后蜱消化细胞ＤＮＡ倍体数显著增大，第５天ＤＮＡ平均倍体数可达６８Ｃ，峰值达１１８Ｃ，≥５Ｃ细胞达３４％。５天后所有消化细胞均≥５Ｃ。中华硬蜱叮咬不同抗原免疫接种兔后中肠消化细胞ＤＮＡ量血餐后不能有效地增加。影响程度依次为柱层析１０５ＫＤ纯化抗原组、中肠抗原组、唾液腺抗原组、全虫抗原组和卵抗原接种组。除卵抗原组外，其它各免疫组平均ＤＮＡ均＜５Ｃ，≥５Ｃ的细胞数＜１７％，峰值＜７Ｃ，且各时期均可见数量不等的２Ｃ和３－４Ｃ细胞，与对照组比较均有显著性差异（Ｐ＜００５）。上述结果提示相关的抗原免疫接种宿主后，可使宿主产生获得性免疫力，导致中华硬蜱吸血后中肠消化细胞ＤＮＡ在血液消化过程中的正常增加受到严重影响，其机理可能与ＤＮＡ合成受阻及消化细胞破坏有关。本实验的五个免疫接种组中以纯化抗原接种组作用最强，在选择制作抗蜱疫苗时可供参考。     

人胎胰腺促性腺激素释放激素免疫反应细胞的形态测量分析

目的: 探讨不同胎龄人胎胰腺GnRH-IR细胞的变化。方法:用免疫组化方法对33例第13周至32周人胎胰腺GnRH-IR细胞显示定位后,用体视学方法对其进行形态学测量分析。结果:(1)GnRH-IR细胞分布于胰岛及外分泌部的腺泡、导管上皮细胞间。(2)人胎胰腺GnRH-IR细胞的数量和大小均随胎龄而增大。胰头和胰尾内、外分泌部GnRH-IR细胞的数密度(Nv)、体密度(Vv)、平均截面面积(A )和平均周长(B)均随胚胎发育而逐渐增大。(3)胰头GnRH-IR细胞的分布密度比胰尾的小。胰头内分泌部和外分泌部GnRH-IR细胞的Vv、Nv比胰尾的小(分别P<0.05和P<0.01=。(4)内分泌部GnRH-IR细胞比外分泌部的大而且密集。相同胎龄组同一部位的内分泌部GnRH-IR细胞的Vv、Nv、A、B参数比外分泌部的要大(分别P<0.05和P<0.01)。结论: 胰腺GnRH-IR细胞存在于人胎胰腺导管、腺泡和胰岛,随胎龄而增多增大,胰头部GnRH-IR细胞比胰尾部少,内分泌部的GnRH-IR细胞比外分泌部更多更大。     

Multicenter retrospective analysis of systemic chemotherapy for advanced neuroendocrine carcinoma of the digestive system

This study analyzed outcomes of systemic chemotherapy for advanced neuroendocrine carcinoma (NEC) of the digestive system. Clinical data from 258 patients with unresectable or recurrent NEC of the gastrointestinal tract (GI) or hepato‐biliary‐pancreatic system (HBP), who received chemotherapy, were collected from 23 Japanese institutions and analyzed retrospectively. Patients had primary sites in the esophagus (n = 85), stomach (n = 70), small bowel (n = 6), colorectum (n = 31), hepato‐biliary system (n = 31) and pancreas (n = 31). Median overall survival (OS) was 13.4 months the esophagus, 13.3 months for the stomach, 29.7 months for the small bowel, 7.6 months for the colorectum, 7.9 months for the hepato‐biliary system and 8.5 months for the pancreas. Irinotecan plus cisplatin (IP) and etoposide plus cisplatin (EP) were most commonly selected for GI‐NEC and HBP‐NEC. For patients treated with IP/EP (n = 160/46), the response rate was 50/28% and median OS was 13.0/7.3 months. Multivariate analysis among patients treated with IP or EP showed that the primary site (GI vs HBP; hazard ratio [HR] 0.58, 95% confidence interval [CI] 0.35–0.97) and baseline serum lactate dehydrogenase levels (not elevated vs elevated; HR 0.65, 95% CI 0.46–0.94) were independent prognostic factors for OS, while the efficacy of IP was slightly better than for EP (HR 0.80, 95% CI 0.48–1.33; P = 0.389). IP and EP are the most common treatment regimens for NEC of the digestive system. HBP primary sites and elevated lactate dehydrogenase levels are unfavorable prognostic factors for survival. A randomized controlled trial is required to establish the appropriate chemotherapy regimen for advanced NEC of the digestive system. This study was registered at UMIN as trial number 000005176.     

全胃切除术后两种消化道重建术式的比较

目的探讨全胃切除术后两种消化道重建术式手术操作时间、病人术后生活质量的影响。方法对2003年1月-2007年6月我院施行全胃切除术的218例病人的临床资料进行回顾性分析。全胃切除后消化道重建分别采用P型空肠袢食管空肠Roux—en—Y吻合术（PR法）（A组,96例）和改良空肠间置代胃吻合术（FJI法）（B组,44例）。记录术中消化道重建时间、术后12个月营养状况和胃肠道症状（GSRS）评分等。结果消化道重建术所需时间两组比较,差异无统计学意义（P〉0．05）。术后12个月B组与A组比较,进食量明显增加（P〈0．05）,而GSRS评分降低（P〈0．05）。术后12个月B组体重恢复较A组更佳,差异有统计学意义（P〈0．05）。结论改良空肠间置代胃吻合术有利于维持病人术后生活质量,在每天进食量及体重恢复方面改良空肠间置代胃吻合术优于P型空肠袢食管空肠Rottx—en—Y吻合术。     

A549细胞系中自噬泡与线粒体形态的初步定量研究

目的探讨发生自噬的细胞中自噬泡的特点及自噬泡与细胞中线粒体形态特征参数间的关系,分析自噬泡膜来源于线粒体膜的可能性。方法将A549细胞培养后收集1～5×106个,常规固定、包埋及超薄切片,在透射电镜下观察。由随机取样的原则选择细胞进行照相。应用CMIAS多功能真彩色图像分析系统测定细胞、线粒体及自噬泡等结构的截面积和截面数目等参数,根据自动测量结果将细胞分为3组。结果 3组细胞内自噬泡总截面面积的平均值明显不同;3组间自噬泡体积密度的平均值之间存在统计学差异(P=0.03)。3组细胞内线粒体数目及线粒体总截面面积的平均值呈相应下降。结论由于3组细胞内线粒体和自噬泡总截面面积的平均值变化趋势相反,推测部分自噬泡的膜可能来源于线粒体的膜。     

上消化道癌前病变患者舌苔与土壤菌群相关性初步分析

目的:探讨扬中地区上消化道癌前病变患者舌苔与土壤菌群的相关性。方法:收集22例上消化道癌前病变患者的舌苔和自留地土壤,采用16S rDNA高通量测序检测菌群,生物信息学分析菌群多样性、菌群结构、共生关系等。结果:舌苔菌群多样性和丰富度均显著低于土壤,菌群组成显著不同。舌苔与土壤之间有17个分类单元（1门、2纲、3目、3科、4属和4种）显著相关。然而,舌苔与土壤菌群预测功能中脂质运输与代谢、翻译后修饰-蛋白翻转-蛋白伴侣、信号转导机制、防御机制等均为显著负相关,胞内运输-分泌-囊泡运输为显著正相关。结论:舌苔与土壤菌群存在相关性,土壤菌群可能是扬中地区上消化道癌地域性高发的关键因素。     

磷酸钙纳米颗粒修饰的甲胎蛋白和树突状细胞疫苗对小鼠皮下移植瘤的抑制作用

目的 探讨转染小鼠带有信号肽的AFP_1 cDNA和去掉信号肽的AFP_2 cDNA树突状细胞(DC)在体外的免疫活性,以及其对Balb/c小鼠皮下移植瘤的抑制作用.方法 应用磷酸钙纳米颗粒将带有信号肽的AFP_1 cDNA和去掉信号肽的AFP_2 cDNA 真核表达载体pcDNA3.1转染DC,将DC疫苗与同源小鼠脾淋巴细胞混合培养,采用酶联免疫吸附(ELISA)法,检测上清液中干扰素γ(IFN-γ)的表达情况.采用四甲基偶氮唑蓝(MTT)法,检测AFP_1/DC和AFP_2/DC刺激同基因小鼠脾淋巴细胞增殖能力及诱导特异性细胞毒性T淋巴细胞(CTL)的杀伤能力.观察AFP_1/DC和AFP_2/DC对Balb/c小鼠皮下移植瘤生长的抑制作用.结果 AFP_2/DC能明显促进脾淋巴细胞增殖并提高CTL的特异性杀伤作用.AFP_1/DC和AFP_2/DC瘤内注射均可显著抑制肝癌移植瘤的生长,但AFP_2/DC的抑制作用更明显,治疗2周后,AFP_2/DC组小鼠肿瘤体积为(726.7±298.2)mm3,明显小于AFP_1/DC组[(1486.2±457.2)mm~3]和空质粒对照组[(2137.2±547.2)mm~3,P<0.05].AFP_2/DC组和AFP_1/DC组的抑瘤率分别达79.2%和39.7%,而空质粒对照组和空白对照组的抑瘤率为0.AFP_2/DC组和AFP_1/DC 组小鼠的生存时间分别为(58.5±4.2)d和(45.2±4.8)d,较空质粒对照组[(30.6±6.2)d]显著延长(P<0.05).结论 AFP_2/DC 疫苗在体外能够诱导出高效而特异的抗肿瘤免疫效应,在体内具有抑制Balb/c小鼠皮下移植瘤生长的作用.     

Thymic nurse cells account for the thymus dependency of preleukemic cells in mice after inoculation of radiation leukemia virus

Inoculation of Radiation Leukemia Virus (RadLV) into C57BL/Ka mice induces thymic lymphomas after a 3-6 month latent period. The leukemogenic process requires a sequence of events from the productive infection of susceptible target cells and induction of preleukemic cells to irreversible neoplastic transformation. Preleukemic cells were detected in the thymus during the first week following virus injection. The thymus dependency of these cells was shown to depend transiently upon peculiar lymphoepithelial complexes called "Thymic Nurse Cells" (TNCs). Indeed, the first preleukemic cells appearing in the RadLV-inoculated thymuses were observed selectively within TNCs. They remained closely associated with these complexes during the first 2 or 4 weeks. Later on, TNCs disappeared almost completely whereas non-TNCs associated preleukemic cells were found. Lymphoepithelial interactions within TNCs were thus required for the initial events of RadLV-induced lymphomagenesis. The subsequent TNCs depletion expressed a disturbance of thymic lymphopoiesis in relation with the neoplastic process.     

Nanomedicine strategies for sustained,controlled, and targeted treatment of cancer stem cells of the digestive system

Cancer stem cells (CSCs) constitute a small proportion of the cancer cells that have self-renewal capacity and tumor-initiating ability. They have been identified in a variety of tumors, including tumors of the digestive system. CSCs exhibit some unique characteristics, which are responsible for cancer metastasis and recurrence. Consequently, the development of effective therapeutic strategies against CSCs plays a key role in increasing the efficacy of cancer therapy. Several potential approaches to target CSCs of the digestive system have been explored, including targeting CSC surface markers and signaling pathways, inducing the differentiation of CSCs, altering the tumor microenvironment or niche, and inhibiting ATP-driven efflux transporters. However, conventional therapies may not successfully eradicate CSCs owing to various problems, including poor solubility, stability, rapid clearance, poor cellular uptake, and unacceptable cytotoxicity. Nanomedicine strategies, which include drug, gene, targeted, and combinational delivery, could solve these problems and significantly improve the therapeutic index. This review briefly summarizes the ongoing development of strategies and nanomedicine-based therapies against CSCs of the digestive system.     

Berberine modulates expression of mdr1 gene product and the responses of digestive track cancer cells to Paclitaxel.

Berberine is the major constituent of Coptis chinese and is commonly used in Chinese herbal medicine to treat patients with gastrointestinal disorders. In this study, using flow cytometry, we have found that a 24-h berberine treatment up-regulated the multidrug-resistant transporter (pgp-170) expression in two oral (KB, OC2), two gastric (SC-M1, NUGC-3) and two colon (COLO 205, CT 26) cancer cell lines. Decreased retention of rhodamine 123 was observed in berberine-treated cells as compared to vehicle control. To examine whether the berberine modulated pgp-170 expression in cancer cells is associated with changes in drug resistance, we determined the cytotoxicity, cell cycle progression and cell morphology of Paclitaxel-treated cells. Paclitaxel (1 nM-10 microM) treatment for 24 h induced cytotoxicity in OC2, SC-M1 and COLO 205 cells in a dose-dependent manner. Pretreatment of cells with 32 microM berberine for 24 h prior to Paclitaxel treatment resulted in increased viability as compared to that of Paclitaxel-treated cells. In addition, Paclitaxel-induced apoptosis and/or G2/M arrest in these three cancer cell lines. Pretreatment of cells with berberine prior to Paclitaxel blocked the Paclitaxel-induced cell cycle responses and morphological changes. These results together suggest that berberine modulated the expression and function of pgp-170 that leads to reduced response to Paclitaxel in digestive track cancer cells.