KGF-transfected cells can stimulate growth and proliferation of human cultured keratinocytes in vitro

Objective: To establish two stably KGF-transfected, immortalized cell lines. Methods: HaCaT-keratinocytes and KMST-6-fibroblasts were transfected by liposome mediated gene transfer. Transfection effectivity, gene integration and configuration of the transgenic protein were investigated by ELISA, DANN-PCR and β-Gal-staining. Results: Most effective GF producing clones were tested by a colorimetric XTT-test. Conclusion: This is a significant acceleration of cell proliferation and mitosis of human keratinocytes in an Air Liquid Interface (ALI) test system.     

大籽獐牙菜中大籽獐牙菜甙的结构

本文报道从大籽獐牙菜Swertia macrosperma C.B.Clark的全草中分离得到的另外四个化合物Ⅰ～Ⅳ。经光谱及化学方法鉴定,化合物Ⅰ是一种咖啡酸双糖酯类化合物,其结构为反式咖啡酸-1-O-芦丁糖酯(trans caffeic acid-1-O-rutinose ester),系首次从天然界得到的一种新化合物,命名为大籽獐牙菜甙(swertiamacroside);化合物Ⅱ～Ⅳ为已知的(口山)酮衍生物,分别鉴定为芒果甙(mangiferin,Ⅱ),bellidifodin(Ⅲ)及bellidifodin-8-O-β-D-glucopyranoside(Ⅳ)。     

大鼠牙胚发育中釉原蛋白的表达

0 引言 牙釉质是由一种特殊的、来源于原始口腔上皮的上皮细胞-成釉细胞分泌的. 牙齿的上皮细胞是以一种特殊的时空方式,分化成有一定功能的成釉细胞,它与来源于外胚间充质的成牙本质细胞的分化有密切的关系[1]. 釉原蛋白是一族基质相关蛋白,由成釉细胞在形成牙釉质时合成和分泌. 釉原蛋白具有非常复杂的生物学功能,迄今为止其确切的功能还不清楚[2],我们对大鼠牙胚发育的不同时期釉原蛋白的表达进行定位,为今后进一步研究釉原蛋白的生物学功能、釉质的生物矿化、细胞的分化奠定基础.     

构建肠系膜微淋巴管内皮细胞培养技术并观察休克淋巴液环境中内皮细胞的形态变化

目的:建立肠系膜微淋巴管内皮细胞的培养技术,观察不同体积分数的休克淋巴液对大鼠肠系膜微淋巴管内皮细胞形态的影响。方法:实验于2004-02/07在河北北方学院病理生理学教研室及实验中心细胞培养室完成。选择健康雄性Wistar大鼠,体质量分别为220～300g和50～80g。实验方法:①无菌条件下制备大鼠重症失血性休克模型,引流肠系膜淋巴液或收集门静脉血;同时另取大鼠,引流正常淋巴液或正常门静脉血。②从动物种类、培养基、植块方法、胎牛血清浓度等方面对植块培养法进行改良,进行肠系膜微淋巴管内皮细胞原代培养并传代。实验分组:分为6组:胎牛血清组:培养液为DMEM 体积分数为0.1的胎牛血清;正常淋巴液组:培养液为DMEM 正常淋巴液;休克淋巴液组:培养液为DMEM 体积分数为0.04,0.06,0.08,0.10的休克淋巴液;正常血浆组:培养液为DMEM 正常血浆;休克血浆组:培养液为DMEM 休克血浆;无血清对照组:培养液为DMEM。实验评估:①光镜下观察大鼠肠系膜微淋巴管内皮细胞原代培养及传代情况。②光镜、扫描电镜及透射电镜下观察不同体积分数的休克淋巴液及不同作用时间(4,8,12h)对肠系膜微淋巴管内皮细胞形态及超微结构的影响。结果:①光镜下肠系膜微淋巴管内皮细胞原代培养及传代情况:内皮细胞呈扁平的梭形或多边形,大小均匀,胞核清晰,呈卵圆形。细胞生长融合形成单层后,呈典型的鹅卵石或铺路石样镶嵌状排列生长。②光镜下在休克淋巴液中肠系膜微淋巴管内皮细胞形态:体积分数为0.04的休克淋巴液作用4h时细胞逐渐收缩、变圆直至脱落漂浮,随着休克淋巴液体积分数增加及作用时间延长,细胞损伤逐渐加重,体积分数为0.08的休克淋巴液作用4h时核旁出现空泡,体积分数为0.10的休克淋巴液作用12h时细胞裂解成碎片。其他各组作用12h肠系膜微淋巴管内皮细胞形态无显著改变。③扫描电镜下在休克淋巴液中肠系膜微淋巴管内皮细胞形态:体积分数为0.04的休克淋巴液作用4h部分细胞逐渐收缩离壁,细胞间隙增大、细胞边缘的突起拉长、缩短、断裂,作用8,12h可见凋亡小体。其他各组作用12h肠系膜微淋巴管内皮细胞形态无显著改变。④透射电镜下在休克淋巴液中肠系膜微淋巴管内皮细胞形态:体积分数为0.04的休克淋巴液作用4h细胞膜形态不规整,胞浆内质网等膜性细胞器扩张,核形态不规则,作用8h线粒体呈球形扩张,细胞核逐渐出现固缩、似细胞凋亡改变,核周腔变宽;体积分数为0.08的休克淋巴液作用时间8h细胞内出现囊泡,内质网不同程度扩张,线粒体主要表现为浓缩、深染等改变,细胞膜边界不清,出现起泡、破碎等现象。其他各组作用12h肠系膜微淋巴管内皮细胞形态无显著改变。结论:成功建立了肠系膜微淋巴管内皮细胞的培养技术;休克淋巴液可导致肠系膜微淋巴管内皮细胞形态及超微结构损伤。     

Efficacy of short‐course lansoprazole with clarithromycin and amoxicillin in the eradication of Helicobacter pylori in South‐East Asian patients: 5‐day t.d.s. versus 7‐day b.d. treatment regimens

OBJECTIVE : To determine and compare the efficacy of 5‐day t.d.s and 7‐day b.d. treatment regimens comprising lansoprazole, clarithromycin and amoxicillin in the eradication of Helicobacter pylori. METHODS : Patients with unequivocal evidence of H. pylori infection based on histology and rapid urease tests of both antrum and corpus biopsies were recruited for the study. The study was a randomized, investigator‐blind, comparative study. Patients received either 500 mg clarithromycin t.d.s. and 500 mg amoxicillin t.d.s. for 5 days (LAC5) or 500 mg clarithromycin b.d. and 500 mg amoxicillin b.d. for 7 days (LAC7) together with 30 mg lansoprazole (both groups) daily for either 5 or 7 days, depending on the treatment group. Patients were assessed for the successful eradication of H. pylori, defined as the absence of bacteria based on histology and urease tests on both antral and corporeal biopsies, carried out at least 4 weeks after completion of the therapy. RESULTS : One hundred and eight patients were recruited for the study. In the LAC5 treatment group, four patients failed to return for follow up and in the LAC7 group, two failed to return for follow up and two were not compliant with medications. Eradication rates based on an intention‐to‐treat analysis were: 46/54 for LAC5 (85.2%; 95% CI = 72.9–93.4) and 47/54 for LAC7 (87.0%; 95% CI = 75.1–94.6). Based on a per protocol analysis, the rates were: 46/50 for LAC5 (92.0%; 95% CI = 80.8–97.8) and 47/50 for LAC7 (94.0%; 95% CI = 83.5–98.7). Both treatment regimens were convenient for patients and except for two patients in the LAC7 group, all patients reported taking 100% of all prescribed medications. The side‐effects encountered were uniformly mild and no patient discontinued treatment because of intolerance to medications. The most common side‐effects were altered taste (LAC5 64.7%; LAC7 78.8%). Diarrhea, nausea and anorexia were reported in a minority of patients. CONCLUSIONS : Both the LAC5 t.d.s. and the LAC7 b.d. treatment regimens were well tolerated by patients and were highly effective in the eradication of H. pylori.     

CD44v6 expression in patients with stage II or stage III sporadic colorectal cancer is superior to CD44 expression for predicting progression

Background: Currently, it is difficult to predict the prognosis of patients exhibiting stage II or stage III colorectal cancer (CRC) and to identify those patients most likely to benefit from aggressive treatment. The current study was performed to examine the clinicopathological significance of CD44 and CD44v6 protein expression in these patients. Study design: We retrospectively investigated 187 consecutive patients who underwent surgery with curative intent for stage II to III CRC from 2007 to 2013 in the Beijing Civil Aviation Hospital. CD44 and CD44v6 protein expression levels were determined using immunohistochemistry and compared to the clinicopathological data. Results: Using immunohistochemical detection, CD44 expression was observed in 108 (57.75%) of the CRC patients; and its detection was significantly associated with greater invasion depth, lymph node metastasis, angiolymphatic invasion, and a more advanced pathological tumor-lymph node-metastasis (TNM) stage. CD44v6 expression was observed in 135 (72.19%) of the CRC patients; and its expression was significantly associated with a poorly differentiated histology, greater invasion depth, lymph node metastasis, angiolymphatic invasion, and a more advanced pathological TNM stage. Expression of CD44v6 was higher than that of CD44 in stage II and stage III sporadic CRC. Conclusion: CD44v6 is a more useful marker for predicting a poor prognosis in stage II and stage III sporadic CRC as compared to CD44.     

体外循环心脏不停跳与非体外循环冠状动脉搭桥术治疗冠状动脉三支病变的疗效

目的 探讨心肺转流(CPB)下行心脏不停跳冠状动脉搭桥术(OPBH)与非体外循环下冠状动脉搭桥术(OPCAB)治疗冠状动脉三支病变的临床效果.方法 210例单纯冠状动脉三支病变患者分为0PCAB组(105例)和OPBH组(105例),分别进行临床观察.结果 OPCAB组无一例需转成OPBH.所有患者均在心脏侧壁和后壁区域搭桥.两组人均冠状动脉远端吻合数和再血管化指数相似;手术后呼吸支持时间、胸腔引流量和输血量OPCAB组明显少于OPBH组;围手术期死亡率和并发症发生率两组比较差异无统计学意义.结论 对于低左室射血分数的患者,虽然出血和输血量较大,术后呼吸支持时间、ICU停留时间较长,但OPBH能取得与OPCAB相同的疗效.