2型糖尿病肾病患者血清中TNF-α、NO与ET的水平变化及其临床意义

目的：探讨2型糖尿病肾病患者血清肿瘤坏死因子仅（TNF-α）、一氧化氮（NO）和内皮素（ET）的水平变化及其临床意义。方法：用ELISA法检测91例2型糖尿病患者血清TNF-α水平，NO与ET的水平分别用硝酸还原酶法和放射免疫法测定。结果：2型糖尿病肾病各组患者TNF-α和ET水平较对照组明显升高，其中ODN组最高（P〈0．01）。而2型糖尿病肾病各组患者血清NO水平较对照组明显减少（P〈0．01）。显性糖尿病肾病患者血清TNF-α和ET呈正相关；血清TNF-α和NO、ET和NO均成负相关。结论：TNF-α、NO与ET可能参与2型糖尿病肾病的发病及病程变化过程，检测患者TNF-α、NO与ET水平可作为判断预后、指导治疗的指标。     

Haploinsufficiency of Pkd2 is associated with increased tubular cell proliferation and interstitial fibrosis in two murine Pkd2 models.

BACKGROUND: Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited human kidney disease and is caused by germline mutations in PKD1 (85%) or PKD2 (15%). It has been estimated that around 1% of tubular cells give rise to cysts, and cell hyperproliferation has been noted to be a cardinal feature of cystic epithelium. Nevertheless, it is uncertain whether the increase in proliferative index observed is an early or late feature of the cystic ADPKD kidney. METHODS: Two Pkd2 mouse mutants (WS25 and WS183) have been recently generated as orthologous models of PKD2. To determine the effect of Pkd2 dosage on cell proliferation, cyst formation and renal fibrosis, we studied renal tissue from Pkd2(WS25/WS25) and Pkd2(+/-) mice by histological analysis. We also examined the proliferative index in archival nephrectomy tissue obtained from patients with ADPKD and normal controls. RESULTS: The proliferative index of non-cystic tubules in Pkd2 mutant mice as assessed by proliferating cell nuclear antigen and Ki67-positive nuclei was between 1-2%, values 5-10 times higher than control tissue. Similarly, the proliferative index of non-cystic tubules in human ADPKD kidneys was 40 times higher than corresponding controls. In Pkd2 mutant mice, significant correlations were found between the fibrosis score and the mean cyst area as well as with the proliferative index. Of significance, proliferating tubular cells were uniformly positive for polycystin-2 expression in Pkd2(+/-) kidney. CONCLUSION: These results suggest that an increase in cell proliferation is an early event preceding cyst formation and can result from haploinsufficiency at Pkd2. The possible pathogenic link between tubular cell proliferation, interstitial fibrosis and cyst formation is discussed.     

环磷酰胺对小鼠肝细胞色素P450含量的影响

目的　了解环磷酰胺 (CP)对小鼠肝细胞色素P4 5 0含量的影响。方法　小鼠腹腔注射CP 2 0mg·kg-1·d-1和CP 4 0mg·kg-1·d-1,连续 4天。以聚乙二醇法制备微粒体 ,测定肝微粒体中P4 5 0的含量。结果　CP 2 0mg·kg-1和CP 4 0mg·kg-1两组动物肝微粒体P4 5 0含量与对照组相比明显下降 (P <0 .0 5 )。结论　CP具有抑制P4 5 0的作用     

抑癌基因P16在膀胱移行细胞癌中的表达及临床意义

目的　探讨抑癌基因P16蛋白在膀胱移行细胞癌中的表达。方法　采用S -P放射免疫法检测 3 2例膀胱移行细胞癌、10例正常膀胱粘膜中P16蛋白的表达。结果　 3 2例膀胱移行细胞癌中 ,P16蛋白表达总阳性率为 40 .63 % ;在膀胱移行细胞癌Ⅰ、Ⅱ、Ⅲ级中分别为 63 .64 %、3 3 .3 3 %、2 5 .0 0 %。P16蛋白表达的阳性率随病理分级增高而降低 ,Ⅰ级与Ⅱ级、Ⅲ级比较差异无显著性 (P >0 .0 5 )。结论　P16蛋白检测有助于对膀胱移行细胞癌的诊断 ;P16蛋白的缺失与膀胱移行细胞癌的发生、发展和预后有关。     

妊娠期糖尿病胎盘细胞凋亡及其与围产结局的关系

目的 :研究胎盘细胞凋亡在妊娠期糖尿病发生及其与临床结局的关系 .方法 :对 4 0例妊娠期糖尿病 (GDM )患者和 4 0例正常对照孕妇的胎盘组织进行检测 ,用DNA缺口原位末端标记 (TUNE)技术检测细胞凋亡 ,电镜观察凋亡细胞的超微结构 ,免疫组织化学方法 (S -P)检测促 (抑 )凋亡基因Bax/Bcl- 2表达水平 .结果 :妊娠期糖尿病组胎盘细胞凋亡指数及Bax、Bcl - 2阳性率、Bax/Bcl- 2的比率均高于对照组 (P <0 0 5 ) ,妊娠期糖尿病组胎盘细胞凋亡指数与孕妇血糖值和新生儿体重呈正相关 ,对照组胎盘细胞凋亡指数与孕妇血糖值和新生儿体重无相关 .结论 :妊娠期糖尿病组和对照组胎盘中均有一定量的细胞凋亡和Bax ,Bcl- 2的表达 ,妊娠期糖尿病组胎盘细胞凋亡明显增加 ,Bax/Bcl- 2的比值增加 .胎盘细胞凋亡及其调节基因的表达间具有一致性 ,妊娠期糖尿病胎盘细胞凋亡增多与不利妊娠结局有关     

具有靶向免疫抑制作用新型重组融合蛋白B7-2-PE40KDEL真核表达载体的构建及其生物效应

目的:构建B7-2-PE40KDEL的真核表达载体,探讨通过体内表达重组融合蛋白,特异性杀伤高表达CD28 T细胞,阻断B7∶CD28/CTLA4共刺激信号途径诱导免疫耐受的可能性.方法:以原核表达载体pRSETA-B7-2-PE40KDEL质粒为模板,采用PCR及酶切连接的方法构建真核表达载体pcDNA3.1/Zeo( )-B7-2-PE40KDEL.利用RT-PCR、Western 印迹及ELISA法从mRNA、蛋白质水平检测了B7-2-PE40KDEL在RPE-CHO真核细胞中的表达情况.采用MTT法对其特异性杀伤高表达CD28 T细胞的生物活性进行测定.结果:成功构建了pcDNA3.1/Zeo( )-B7-2-PE40KDEL真核表达载体,转染入RPE-CHO细胞后可转录翻译为重组融合毒素蛋白,相对分子质量约为71×103,平均106个转染细胞24 h表达0.23 μg/L.活性测定显示真核载体所表达的B7-2-PE40KDEL可特异性地抑制高表达CD28的人T淋巴细胞系,而对CD28阴性的人白血病细胞系的抑制率较低.结论:真核载体表达的B7-2-PE40KDEL具有良好的靶向性免疫抑制活性,为进一步开展其体内特异性防治移植排斥反应及自身免疫性疾病奠定了基础.     

重组骨形态发生蛋白-2结合软骨下钻孔治疗犬关节软骨全层缺损的实验研究

目的 研究重组骨形态发生蛋白-2(rhBMP-2)结合软骨下骨钻孔治疗犬关节软骨全层缺损的可行性，为临床应用提供实验依据。方法 依照软骨缺损处理方法的不同将64侧股骨髁随机均分为4组：①结合组：软骨下骨钻孔 胶原海绵吸附rhBMP-2充填软骨缺损；②BMP组：胶原海绵吸附rhBMP-2充填软骨缺损；③钻孔组：单纯软骨下骨钻孔；④对照组：不作处理或单纯用胶原海绵填塞。术后2、4、8、12周取材观察其大体、光镜、透射电镜、免疫组织化学情况。结果 除对照组仅有纤维组织修复外，其余3组均有不同程度的软骨修复，但结合组的修复在组织细胞形态、超微结构、Ⅱ型胶原含量等方面均明显优于其他两组。结论rhBMP-2结合软骨下骨钻孔能有效修复犬膝关节软骨的全层缺损，该技术可行，有望在临床应用。     

Ki-67、增殖细胞核抗原、P53在脑膜瘤中的表达及意义

目的 :通过对 Ki- 6 7、增殖细胞核抗原、P5 3在脑膜瘤中的表达 ,进一步研究其与肿瘤的良、恶性及肿瘤复发的关系 ,从而更好判断其生物学行为。方法 :行脑膜瘤全切除手术治疗 ( Simpson's Grade or Grade )并经 9～12 4个月 ( 4 8± 9)随访的 6 0例 ,采用 SP免疫组化法 ,检测手术切除石蜡包埋的肿瘤标本的 Ki- 6 7、增殖细胞核抗原、P5 3抗原 ,以显微镜下的细胞核内出现棕色颗粒为阳性 ,计算 3者的标记指数 ( L abeling Index) .对患者的临床资料与肿瘤细胞的标记指数的差异性进行统计学分析。结果 :Ki- 6 7、增殖细胞核抗原、P5 3在不同病理形态脑膜瘤中的表达具有显著性差异 ( P<0 .0 5 ) ,恶性脑膜瘤、非典型性脑膜瘤中的表达明显高于良性脑膜瘤 ;同样在复发性脑膜瘤中其表达也明显高于非复发性脑膜瘤 ( P<0 .0 5 ) ;良性脑膜瘤中 Ki- 6 7、增殖细胞核抗原的表达在复发组明显高于非复发组 ( P<0 .0 5 ) ,而 P5 3的表达在复发组与非复发组无统计学意义 ( P>0 .0 5 )。结论 :( 1) Ki- 6 7、增殖细胞核抗原、P5 3的表达与病理分型和脑膜瘤术后复发有关。 ( 2 )良性脑膜瘤结合 Ki- 6 7、增殖细胞核抗原的表达情况可用来判断预后即有无复发的可能 ,从而进一步来指导临床治疗     

血管平滑肌细胞凋亡及Bcl-2和Bax蛋白表达在自发性高血压大鼠颈动脉血管间质胶原重构及逆转中的意义

目的　探讨血管平滑肌细胞及Bcl 2和Bax蛋白表达在自发性高血压大鼠 (SHR)颈动脉血管间质胶原重构及逆转中的意义。方法　选择 16周龄雄性SHR各 12只分别为厄贝沙坦组 (30mg·kg- 1·d- 1 )和非厄贝沙坦组 ,另选同龄雄性Wistar大鼠 12只为正常对照组 ;颈总动脉切片采用HE染色、苦味酸一天狼星红 (PSR)染色、免疫组化法染色和末端脱氧核苷酸转移酶介导dUIP缺口末端标记法分别检测颈动脉壁厚 腔径比值、胶原面积百分比、Bcl 2和Bax蛋白表达和血管平滑肌细胞 (VSMC)凋亡率(APOI)。结果　非厄贝沙坦组颈动脉壁厚 腔径比值较正常对照组高 2 5 0 .11%± 15 .6 3% (P <0 .0 1) ,厄贝沙坦组治疗 2 0周后 ,较非厄贝沙坦组下降了 4 3.2 4 %± 8.6 2 % (P <0 .0 1) ;非厄贝沙坦组胶原面积百分比较正常对照组SBP高 2 0 1.6 7%± 16 .73% (P <0 .0 1) ,厄贝沙坦组治疗 2 0周后 ,较非厄贝沙坦组仅仅下降了 17.70 %± 6 .4 5 % (P <0 .0 1) ;非厄贝沙坦组的VSMC率APOI较正常对照组显著增高 ,经药物治疗后下降 ;非厄贝沙坦组的VSMCBcl 2蛋白表达阳性率明显高于正常对照组 ,用厄贝沙坦治疗后 ,VSMCBcl 2阳性率较非厄贝沙坦组明显减少 ;非厄贝沙坦组和正常对照组的VSMCBax蛋白表达阳性率无明显差别 ,用厄贝沙坦治疗后 ,VS     

An animal model of chronic inflammatory pain: pharmacological and temporal differentiation from acute models.

Clinically, inflammatory pain is far more persistent than that typically modelled pre-clinically, with the majority of animal models focussing on short-term effects of the inflammatory pain response. The large attrition rate of compounds in the clinic which show pre-clinical efficacy suggests the need for novel models of, or approaches to, chronic inflammatory pain if novel mechanisms are to make it to the market. A model in which a more chronic inflammatory hypersensitivity phenotype is profiled may allow for a more clinically predictive tool. The aims of these studies were to characterise and validate a chronic model of inflammatory pain. We have shown that injection of a large volume of adjuvant to the intra-articular space of the rat knee results in a prolonged inflammatory pain response, compared to the response in an acute adjuvant model. Additionally, this model also results in a hypersensitive state in the presence and absence of inflammation. A range of clinically effective analgesics demonstrate activity in this chronic model, including morphine (3mg/kg, t.i.d.), dexamethasone (1mg/kg, b.i.d.), ibuprofen (30mg/kg, t.i.d.), etoricoxib (5mg/kg, b.i.d.) and rofecoxib (0.3-10mg/kg, b.i.d.). A further aim was to exemplify the utility of this chronic model over the more acute intra-plantar adjuvant model using two novel therapeutic approaches; NR2B selective NMDA receptor antagonism and iNOS inhibition. Our data shows that different effects were observed with these therapies when comparing the acute model with the model of chronic inflammatory joint pain. These data suggest that the chronic model may be more relevant to identifying mechanisms for the treatment of chronic inflammatory pain states in the clinic.