miR-548d抑制骨肉瘤细胞的增殖和迁移

目的:探讨miR-548d对骨肉瘤细胞增殖和迁移的影响。方法:通过Real time PCR及Western blot检测miR-548d和KRAS在30例骨肉瘤组织以及293、MG63和U2OS细胞中的表达情况。对30例骨肉瘤组织中miR-548d和KRAS含量的相关性进行分析，随后通过报告基因实验印证miR-548d对KRAS的靶向作用。MG63细胞中分别过表达或沉默miR-548d后，通过Western blot实验分析KRAS的变化情况，MTT实验观察miR-548d对细胞增殖的影响，Transwell实验观察miR-548d对其迁移能力的影响。结果:骨肉瘤组织及细胞中miR-548d表达较低，KRAS表达较高。在骨肉瘤组织中miR-548d与KRAS的表达呈负相关。报告基因实验证明miR-548d可以直接打靶KRAS。Western blot指出miR-548d可以抑制KRAS的表达。最后MTT和Transwell实验指出miR-548d可以抑制MG63细胞的增殖与迁移。结论:miR-548d可以通过打靶KRAS抑制骨肉瘤细胞的增殖和迁移。     

P63阳性基底细胞在前列腺切除术后尿道再上皮化中的作用

目的:探讨前列腺切除术后P63阳性基底细胞在前列腺部尿道上皮修复过程中的作用。方法:15只老年家犬行前列腺增生切除术,术后第3、7、14天取材行HE染色和免疫组织化学染色分别观察前列腺部尿道上皮修复过程和P63蛋白表达水平。结果:术后3 d手术创面偶见P63阳性细胞;术后7 d可见片状P63阳性细胞,且这些细胞与残余前列腺组织相连续;术后14 d可见新生尿道上皮基底层细胞阳性表达P63。结论:残余前列腺组织是BPH术后尿道再上皮化的重要组织,而位于基底层的P63阳性细胞可能是这一修复过程的源细胞之一。     

枸杞多糖对大鼠角膜缘干细胞体外增殖的影响研究

目的通过体外分离和培养大鼠角膜缘干细胞(Limbal Stem Cells, LSCs)，观察枸杞多糖(Lyciumbarbarum Polysaccharide, LBP)对LSCs体外增殖的影响，为中药体外培养LSCs的研究提供实验资料。 方法体外分离和培养LSCs，显微镜观察LSCs的细胞形态特征，通过免疫荧光检测转录因子p63初步鉴定大鼠LSCs；分对照组和枸杞多糖处理组，每组20例，免疫印迹检测增殖细胞核抗原(Proliferating Cell Nuclear Antigen, PCNA)，初步探讨枸杞多糖对LSCs的增殖影响。 结果镜下观察LSCs体积小，呈多边形或椭圆形，胞核较大，核质比例大，免疫荧光结果显示体外分离的LSCs可表达p63(阳性率为76.41±1.66%)；枸杞多糖处理后LSCs的PCNA蛋白的表达水平高于对照组，并且随着枸杞多糖浓度的增加，PCNA蛋白的表达升高(P<0.05)。 结论枸杞多糖对LSCs的增殖和生长具有促进和维持的作用。     

泛素蛋白酶体抑制剂MG-132与心力衰竭

心力衰竭是各种心脏疾病发展的严重阶段,发病率的上升是心血管医学面临的最大挑战之一,已成为心脏病治疗的最后战场。MG-132是一种泛素蛋白酶体抑制剂,其在心力衰竭发展过程中扮演重要的角色,但其具体作用尚未完全阐明。现综述MG-132与心力衰竭关系及相关分子机制。     

MicroRNA-301b-3p accelerates the growth of gastric cancer cells by targeting zinc finger and BTB domain containing 4

MicroRNAs (miRNAs) have been found to be aberrantly expressed and exert essential roles in the tumorigenesis and progression of gastric cancer (GC). miR-301b-3p has been recognized as a cancer-related miRNA in lung cancer, bladder cancer and hepatocellular carcinoma. However, the function of miR-301b-3p in GC progression and its underlying mechanism have not been studied yet. In this study, we found that miR-301b-3p expression was up-regulated in GC tissues compared to adjacent noncancerous tissues. Furthermore, the elevated levels of miR-301b-3p were detected in GC cell lines (SGC-7901, AGS, MKN-45 and MGC-803) as compared with GES-1 cells. Interestingly, GC tissues from patients with tumor size ≥ 5 cm and advanced tumor stages showed obvious higher levels of miR-301b-3p compared to matched controls. Functionally, miR-301b-3p knockdown prominently inhibited cell proliferation, and induced cell cycle arrest at G1 phase and apoptosis in MGC-803 cells. Meanwhile, ectopic expression of miR-301b-3p conversely regulated these biological behaviors of MKN-45 cells. Next, we found that miR-301b-3p knockdown increased, whereas miR-301b-3p overexpression reduced the expression of zinc finger and BTB domain containing 4 (ZBTB4) in GC cells. Accordingly, luciferase reporter assay identified ZBTB4 as a direct target of miR-301b-3p. ZBTB4 overexpression markedly restrained the growth of MGC-803 cells. More importantly, ZBTB4 silencing partially reversed miR-301b-3p knockdown-induced tumor suppressive effects on MGC-803 cells. In conclusion, we firstly revealed that miR-301-3p was highly expressed in GC and contributed to tumor progression via attenuating ZBTB4, which might provide a novel molecular-targeted strategy for GC treatment.     

Immunohistochemical analysis of c-erbB-2, Bcl-2, p53, p21WAF1/Cip1, p63 and Ki-67 expression in hydatidiform moles

Hydatidiform moles (HM) are characterized by an abnormal proliferating trophoblast with a potential for a malignant transformation. Similar to other human tumors, trophoblastic pathogenesis is likely a multistep process involving several molecular and genetic alterations. The study was performed to investigate the expression patterns of c-erbB-2 and Bcl-2 oncoproteins, p53, p21^WAF1/CIP1 and p63 tumor suppressor proteins and Ki-67 cell proliferation marker in HM.We conducted a retrospective study of 220 gestational products, including 39 hydropic abortions (HA), 41 partial HM (PHM) and 140 complete HM (CHM). The expression of c-erbB-2, Bcl-2, p53, p21^WAF1/CIP1, p63 and Ki-67 was investigated by immunohistochemistry on archival tissues. c-erbB-2 expression was observed in three PHM and 10 CHM. Bcl-2 immunostaining was significantly higher in PHM (61%) and CHM (70.7%) compared with HA (7.7%, p?=? 0.001 and p?<? 0.0001, respectively). p53 expression was stronger in CHM (73.6%) compared with PHM (24.4%, p < 0.0001) and HA (12.8%, p < 0.0001). p21^WAF1/CIP1 staining was observed as well in molar and non-molar gestations (p?>? 0.05). p63 immunoexpression was significantly described in CHM (85.7%) and PHM (78%) compared with HA (10.2%, p < 0.0001 and p = 0.0001, respectively). Ki-67 was significantly expressed in CHM (72.1%) compared with HA (46.2%, p = 0.005).Altered expression of Bcl-2, p53, p63 and Ki-67 reflects the HM pathological development. Immunohistochemical analysis is beneficial to recognize the HM molecular and pathogenic mechanisms. Furthermore, it could serve as a useful adjunct to conventional methods for refining HM diagnosis.