Wistar大鼠胰岛细胞体外分离、纯化及鉴定

目的 探索Wistar大鼠胰岛分离纯化的最佳条件.方法 采用肝胰管内灌注胶原酶消化分离胰岛及Ficoll 400密度梯度离心纯化.纯化后的胰岛经组织学染色、电镜及放射免疫法鉴定其特异性和活力.结果 组织学染色显示纯化后胰岛的活力和纯度分别在95%和85%以上.电镜显示纯化后的胰岛形态完整,包膜清晰,分泌颗粒丰富.放射免疫结果表明,低糖组和高糖组分泌胰岛素的浓度有显著差异,证明胰岛功能良好.结论 肝胰管内灌注胶原酶消化法是一种好的消化方法.影响胰岛收获量的因素很多,例如胰腺的充分扩张,胶原酶的浓度和活性以及消化的时间等.     

Decreased mtDNA Copy Number of Gastric Cancer： a New Tumor Marker？

Objective To explore the relationship between mtDNA (mitochondnal DNA) and gastric cancer by comparing the difference of mtDNA copy number in gastric cancers and paracancerous tissues. Methods The HV1 (hypervariable region) and HV2 of the mitochondrial D-loop region from 20 cases of gastric cancer and 20 paracancerous tissues were amplified by PCR with βactin serving as a quantitative standard marker. The products were separated by polyacrylamide gel electrophoresis (PAGE) and silver stained in order to compare the difference in mtDNA copy number between gastric cancers and paracancerous tissues. The mtDNA copy number was determined for gastric cancers having various pathological characteristics and the results compared with previous immunohistochemical staining of the tumors. Results There was a significantly quantitative difference in HV1, HV2 (standardized with βactin) between gastric cancers and paracancerous tissues (P<0.01 ). The mtDNA copy number was associated with important enzymes in the nucleus such as AKP, cAMP-PDE and cGMP-PDE (P< 0.05), but not associated with tumor histological type and invasive depth (P> 0.05). Conclusion The occurrence of gastric cancer was closely associated with decreased mtDNA copy number, which may be a new tumor marker. This work was supported by the research grants from the National Natural Science Foundation of China (No. 30371607).     

Ability of a Specific ERK Signal-Pathway Inhibitor to Reverse P-Glycoprotein- Mediated Vincristine Resistance in Colon Cancer Cell Unes

OBJECTIVE To investigate the effect of a specific inhibitor PD098059 of the extracellular-signal regulated protein kinase (ERK) pathway on the P-glycoprotein (P-gp)-mediated resistance of colon cancer cell lines SW480/VCR and CoLo205NCR.METHODS SW480NCR and CoLo205NCR cells were generated byexposuring SW480 and CoLo205 cells to vincristine (VCR) (30 ng/ml) for 72h, which resulted in a comparatively higher level of P-gp expression.Western blotting was used to analyze P-gp, MRP, LRP, GST-‘rr and TOPOIIexpression after exposuring the SW480 and CoLo205 cells to VCR (30 ng/ml)for 72 hrs. P-gp and pERK1/2 expressions was analyzed in SW480NCR andCoLo205/VCR cells treated with or without the specific inhibitor of MEK,PD098059. The MTT assay was used to determine the susceptibility ofSW480NCR and CoLo205NCR cells to VCR, treated with or withoutPD098059.I~F.SULI“S The results showed that VCR induced a comparatively higher levelof P-gp expression in the cell lines, but not that of MRP, LRP, GST-n- orTOPOII. P-gp expression levels were depressed significantly in SW480/VCR and COLO205/VCR cells by the specific inhibitor of MEK, PD098059.The IC50 (248 19.6 and 215 10.7 ng/ml) to VCR of SW480/VCR andCoLo205/VCR cells exhibited a 2.16 and 2.03-fold higher resistancecompared to the negative control group (SW480 and CoLo205 cells)(115 15.6 and 106 11.9 ng/ml), but a 1.35 and 1.21 -fold higher resistance thanthe group treated with VCR (30 ng/ml) PD098059 (184 21.8 and 177 19.4 ng/ml).CONCLUSION This study shows that the expression of P-gp can beinduced by exposuring cells to VCR, and that this induction can be reversedby inhibiting the ERK signaling pathway at the point of MEK by its specificinhibitor, PD098059. The ERK signal-transduction pathway may play a rolein modulating mdrl expression in colon cancer.     

mRNA EXPRESSION OF PTEN AND VEGF GENES IN EPITHELIAL OVARIAN CANCER

VEGF gene, also known as vascular permeability factor, has been mapped to chromosome 6p21.3[11]. Biochemically, it is a heparin binding glycoprotein that has in at least four molecular isoforms, among which, VEGF165 occurs most common. VEGF can act as a specific mitogen for a variety of endothelial cells in vitro and as an angiogenic molecule in vivo [11,12]. VEGF is a potent multifunctional cytokine that exerts several potentially independent actions on the vascular endothelium, includin…     

STUDIES OF THE CHARACTERISTIC FEATURES OF KI-1 POSITIVE NON-HODGKIN'S LYMPHOMA

Ki-lantigen(CD3o)waslO5oOo/12oooOglycoproteindistinguishedbymonoclonalantibodyKi-lwhichwasmadefromHodgkin'sdisease(HD)cellsstrainL-428,'eXPressedonHDcellsandReed-Sternberg(R-S)cellsatfirstthoughtItwasfoundbySteinandothersinl985,whonamedKi-lstrongpositiveNHLasKi-llymphoma,thatKi-lantigenwasalsoexpressedonsomeNHL.2Theresearchesonthisspecialtypeoflymphomahavealreadyappearedabroad.WehavecollectedfivecasesofKi-lpositiveNHLandnowdiscusstheirclinicalhistopathologicalandimmunophenotypicfe…     

LOSS OF HETEROZYGOSITY ON CHROMOSOME 13 IN SQUAMOUS CELL CARCINOMAS OF THE LARYNX

ＬＯＳＳＯＦＨＥＴＥＲＯＺＹＧＯＳＩＴＹＯＮＣＨＲＯＭＯＳＯＭＥ１３ＩＮＳＱＵＡＭＯＵＳＣＥＬＬＣＡＲＣＩＮＯＭＡＳＯＦＴＨＥＬＡＲＹＮＸＢａｉＳｕｊｕａｎ白素娟ＺｈａｎｇＸｕｅ张学ＷａｎｇＪｕｎ王筠ＳｕｎＫａｉｌａｉ孙开来ＦｅｉＳｈｅｎｇｚｈｏｎ...     

吲哚美辛对5-氟尿嘧啶在大鼠体内药代动力学的影响

目的探讨吲哚美辛(Ind)增强5-氟尿嘧啶(5-Fu)的抗肿瘤作用是否与药代动力学的变化有关。方法采用反相高效液相色谱(HPLC)法测定大鼠血浆中5-Fu浓度,数据用“3p97”程序处理。结果单用和联用Ind后,5-Fu在大鼠体内的药代动力学均为一室模型,t1/2ka分别为(3.8±2.0)和(4.4±2.7)min,t1/2ke分别为(57±46)和(101±48)min,ke分别为(0.018±0.010)和(0.009±0.005)min-1,cmax分别为(7.0±1.8)和(19±8)mg.L-1,AUC分别为(0.7±0.5)和(3.5±2.4)g.min.L-1,Cl分别为(100±50)和(24±21)mg.kg-1.min-1。说明联用Ind后,5-Fu的作用时间延长,在大鼠体内的经时过程增长。结论Ind能延缓5-Fu在大鼠体内的消除,两者联用时应注意调整给药剂量和对5-Fu进行临床给药监测。     

切除部分腭咽肌的改良UPPP术后疗效观察

目的:为了提高改良UPPP术后疗效,术中切除部分腭咽肌,并对术后疗效、咽腔成形特点及可能出现的并发症等进行临床观察。方法:选择经Apneagraph睡眠呼吸监测阻塞定位仪(AG)、纤维鼻咽喉镜结合Müller检查及鼻咽部3D-CT确诊的OSAHS患者82例,随机分为对照组26例,于全身麻醉下行传统的改良UPPP术,术中不切除部分腭咽肌;实验组56例,于全身麻醉下行改良UPPP术,术中切除部分腭咽肌。术后半年内每个月通过复查或电话随访的方式了解患者是否出现鼻腔反流、耳闷及听力下降等并发症。术后6个月采用嗜睡程度评估量表(ESS)评价嗜睡状态,应用t检验与术前ESS评分进行统计学对比分析。术后6个月应用AG评价疗效。通过测量悬雍垂长度(L1)、软腭游离缘与咽后壁距离(L2)及鼻咽峡宽度(L3)了解咽峡成形结构特点,应用多元线性回归分析进行假设检验并评价各测量值与疗效的关系。用SPSS19.0软件进行数据分析。结果:术后6个月实验组显效50例(89.29%),有效6例(10.71%);术前ESS评分11.74±2.48,术后3.84±2.05。对照组显效19例(73.08%),有效7例(26.92%);术前ESS评分11.91±2.40,术后6.92±2.47,两组疗效差异有统计学意义(P<0.01)。术后半年内两组均未见耳闷、听力下降、自家音增大等反映咽鼓管功能障碍的并发症,半年后两组咽腔功能均恢复正常。术后6个月实验组和对照组L1分别为(5.91±3.38)mm和(6.20±3.76)mm(P>0.05);L2分别为(15.70±3.29)mm和(15.35±1.44)mm(P>0.05);L3分别为(20.54±3.33)mm和(16.43±2.21)mm(P<0.05),说明实验组的术后L3明显增宽。通过多元线性回归分析,术后疗效与L2手术前后的差值和L3手术前后的差值有线性回归关系,并与二者呈负相关。由标化回归系数看出,L3手术前后的差值对术后疗效影响最大。结论:切除部分腭咽肌的改良UPPP通过有效增加术后鼻咽峡宽度提高了手术疗效,并且术后未出现咽鼓管功能、软腭功能、吞咽功能及构音功能障碍。