信号通路在肝癌发生过程中的作用及靶向治疗策略

肝癌是一种多病因参与、多阶段病变的复杂疾病。研究表明,参与调节机体重要生理过程的一些信号通路异常与癌变密切相关。该文综述了在肝癌发生过程中一些信号通路的异常现象,总结并分析了以信号通路为靶点的临床药物研究策略及未来发展方向。对这些信号通路作用机制进行深入细致了解将有助于寻找更为合适而有效的治疗靶点。     

PI3K/Akt信号转导通路在ALV-J感染中作用的初步研究

 【目的】探讨ALV-J在宿主细胞中复制与PI3K/Akt信号转导通路的关系。【方法】将血管瘤病变型ALV-J毒株HN06和骨髓瘤病变型ALV-J毒株NX0101分别感染DF-1细胞,通过Western blot、Real-time PCR、IFA和ELISA等方法,观察细胞Akt蛋白磷酸化水平、病毒RNA表达水平和病毒蛋白表达水平等指标。【结果】HN06株和NX0101株在体外细胞中复制水平有差异。HN06株的早期感染可引起Akt转导通路的活化,病毒引起的Akt磷酸化具有病毒滴度依赖性,而且能被PI3K特异性抑制剂LY294002所抑制,表明HN06株诱导的Akt活化是PI3K途径依赖的。LY294002可在病毒感染早期呈剂量依赖性地显著降低受染细胞中HN06 RNA水平、囊膜蛋白水平和细胞培养物上清中的病毒粒子含量。【结论】PI3K/Akt信号转导通路活化对HN06株在细胞感染早期具有重要的作用,该结果与已报道的有关细胞PI3K/Akt信号转导通路参与NX0101株的早期感染的结论一致。本研究为进一步阐明ALV-J入侵宿主细胞和复制的精确机制等研究奠定了基础。     

Nitric oxide impacts bovine sperm capacitation in a cGMP‐dependent and cGMP‐independent manner

We aimed to elucidate whether NO acts in in vitro sperm capacitation in bovine via cGMP/PKG1 pathway. For this, cryopreserved bovine sperm were capacitated in vitro with 20 µg/ml heparin (Control) plus treatments: 1 mM L‐arginine (L‐arg, NO precursor), 50 µM Rp‐8‐Bromo‐β‐phenyl‐1,N²‐ethenoguanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐Br‐cGMPS, selective inhibitor of the binding site for cGMP in PKG1), 1 mM 2‐Phenyl‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl 3‐oxide (PTIO, NO scavenger), and the combinations of L‐arg + RP‐8‐Br‐cGMPS and L‐arg + PTIO. Sperm motility and vigour were determined by phase‐contrast microscopy, capacitation status by chlortetracycline staining, and the intracellular concentration of cGMP was measured by ELISA. Data were subjected to analysis of variance and means compared with SNK test at 5% probability. Motility and vigour were lower in sperm treated with PTIO when compared to Control and other treatments (p < .05). The L‐arg treatment showed the highest percentage of capacitated sperm when compared to the Control and other treatments (Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO) (69.8 ± 3.4%, 51.2 ± 3.0, 51.1 ± 2.1, 51.2 ± 3.0 and 45.5 ± 2.7, respectively) (p < .05). The capacitation ratio (%) was lower in treatments with Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO, respectively (p < .05). Lastly, cGMP concentration (pmol/ml) was lower in PTIO and L‐arg + PTIO (1.3 ± 0.3 and 1.6 ± 0.4) and was higher in Rp‐8‐Br‐cGMPS and L‐arg + Rp‐8‐Br‐cGMPS (3.7 ± 0.4 and 4.0 ± 0.5) treatments. We showed that during in vitro capacitation of cattle: (a) NO influences sperm motility and vigour; (b) NO is associated with cGMP synthesis through two independent pathways and (c) the cGMP/PKG1 pathway has a partial role in sperm capacitation and does not involve the L‐arg/NO.     

葡萄果实中莽草酸途径与多酚积累的关系

概述了莽草酸途径及其在植物次生代谢中的重要作用、葡萄果实类黄酮代谢与调控的研究现状,并对莽草酸代谢与葡萄多酚积累的关系进行探讨和展望。     

CD36/Src/ERK pathway is involved in process of monocyte differentiation into macrophages

AIM To investigate the role of fatty acid translocase (FAT/CD36) on differentiation of monocytes to macrophages. METHODS Human monocyte THP-1 cells were treated with phorbol 12-myristate 13-acetate (PMA) at 0, 100 and 200 μg /L. Small interfering RNA (siRNA) targeting CD36 (siCD36) was employed to knock down the expression of CD36 in THP-1 cells. The CD36 over-expression (CD36OE) cell line was constructed by transfection with a recombinant lentivirus containing CD36 cDNA. Optical microscopy and crystal violet staining were used to detect the monocyte morphological changes and adhesion ability. The protein expression of CD36 was measured by flow cytometry and Western blot. The mRNA levels of CD36, CD11b and CD80 were detected by real-time PCR. The protein levels of extracellular signal-regulated kinase (ERK） and Src tyrosine kinase were determined by Western blot. RESULTS The cellular adhesiveness of THP-1 cells was elevated in the process of monocytes differentiation, and the expression of CD36 was increased in this process as well (P<0.01). siCD36 was transfected into the THP-1 cells (CD36i group) and the silencing efficiency was approximately 80%. The cell surface area and cellular adhesiveness were significantly decreased in CD36i group compared with scrambled siRNA (NCi) group (P<0.01). The mRNA levels of CD11b and CD80 were decreased in CD36i group compared with NCi group (P<0.01). The cell surface area and cellular adhesiveness were increased in CD36OE group compared with empty vector (vector) group (P<0.05). The mRNA levels of CD11b and CD80 were increased in CD36OE group compared with vector group (P<0.01). The phosphorylation levels of ERK and Src were decreased in CD36i group compared with NCi group (P<0.05). CONCLUSION CD36 promotes the differentiation of human monocyte THP-1 cells to macrophages by increasing the phosphorylation of Src and further activating ERK.     

花粉管通道法转基因抗虫棉外源基因的整合方式

对花粉管通道法培育的转Bt基因抗虫棉GK19、GK22、1016、1018、1022、1025、GK5、GKsu12和SGK1进行Southern杂交分析。结果表明,目的基因能够完整整合到基因组里,HindⅢ酶切位点有可能发生甲基化或丢失。除1018以外的上述8个抗虫棉的分析结果还表明,载体骨架没有整合到棉花基因组中。     

Mechanism of JAK2-STAT3 signaling pathway in ischemic postconditio-ning neuroprotection after cerebral ischemia in tree shrews

AIM: To investigate the regulatory effect of JAK2-STAT3 signaling pathway on the neuroprotection of ischemic postconditioning (IPoC) in tree shrews, and to explore the mechanisms of cerebral injury deterioration after inhibiting the JAK2-STAT3 pathway. METHODS: The model of thrombotic cerebral ischemia was induced by photochemical reaction in tree shrews and the IPoC was established at 4 h after ischemia followed by clipping ipsilateral common carotid artery on the ischemia side for 5 min (3 times). After IPoC and intracerebroventricular injection of AG490 (JAK2 inhibitor), the changes of cerebral infarction area were detected by TTC staining, and the histological and ultrastructural changes of cortical neurons were observed under light and electron microscopes, respectively. The protein levels of t-STAT3 and p-STAT3 in the cortical tissue were determined by Western blot. RESULTS: The neuronal pycnosis, mitochondrial swelling and vanish of the mitochondrial cristae were found in cortical cortex, and the infarction area was (24.78±3.30)% at 24 h after cerebral ischemia. Meanwhile, the phosphorylation level of STAT3 protein in the cortical tissue was significantly increased (P<0.01). The cortical neuronal damage and mitochondrial swelling were decreased after IPoC, the area of cerebral infarction was significantly reduced to (17.67±1.83)% (P<0.01), and the phosphorylation level of STAT3 protein was further increased (P<0.01). However, the neuronal damage was aggravated, the infarction area was expanded to (23.85±2.77)%(P<0.05) after treatment with AG490, and the phosphorylation level of STAT3 protein was also significantly reduced (P<0.05). CONCLUSION: IPoC may reduce cerebral injury by regulating the phosphorylation of STAT3 protein, and inhibition of JAK2-STAT3 signaling pathway may counteract the cerebral protective effect of IPoC and aggravate brain injury.     

一株乙草胺降解菌株M-3的分离鉴定及其代谢途径的初步研究

乙草胺是一种广谱、高效的酰胺类除草剂,由于乙草胺具有较长的降解周期,还有不易挥发、不易光解、易残留的特点,如果过量、频繁使用,对人、动植物均有一定的毒害作用。为了探讨乙草胺的微生物降解机理,本研究从长期受乙草胺污染的土壤中分离到一株能降解乙草胺的菌株M-3,该菌株能以乙草胺为唯一碳源生长。通过菌落表型、生理生化特征和菌株16SrRNA基因序列的相似性分析,将其鉴定为寡养单胞菌属(Stenotrophomonas sp.)。在室内条件下,运用HPLC和HPLC-MS分析方法,研究了菌株M-3对乙草胺的降解特性,并对其代谢途径做了初步的研究。结果表明,菌株M-3在5d内对浓度为50mg/L的乙草胺的降解率可达76.6%。M-3降解乙草胺的最适温度和最适pH值分别为30℃和7.0。在对代谢产物结果进行分析的基础上推测了M-3降解乙草胺的途径,将产物鉴定为2-乙基-6-甲基-2-氯乙酰苯胺。该研究为乙草胺污染的生物修复提供了理论依据。     

基于REE示踪的土壤团聚体破碎转化路径定量表征

土壤团聚体破碎转化路径是坡面侵蚀过程研究的难点问题之一。目前团聚体破碎转化路径的定量表征仍不明晰,一定程度上限制了土壤侵蚀过程中泥沙分选搬运机制的深入研究。基于大样带调查选取6种不同质地的典型农耕地土壤为研究对象,结合稀土元素(Rare Earth Elements,REE)示踪方法,综合分析不同粒径土壤团聚体(5~2,2~1,1~0.5,0.5~0.25,<0.25 mm)和不同径流扰动周期(24 h,7天)对REE吸附和解吸能力的影响,探究REE示踪不同粒径土壤团聚体破碎转化的可行性,定量表征了土壤团聚体破碎转化路径。结果表明:REE与土壤团聚体的实际吸附浓度低于施放浓度,2~1,1~0.5,0.5~0.25,<0.25 mm土壤团聚体的REE吸附浓度与黏粒含量呈显著正相关(P<0.05);径流扰动影响对吸附于土壤团聚体的REE解吸作用十分微弱,解吸浓度仅占REE实际吸附浓度的0.001%~0.139%。5~2,2~1,1~0.5,05~0.25,<0.25 mm土壤团聚体经过湿筛后向各粒径转化的路径基本相同,向<0.25 mm微团聚体转化为土壤团聚体破碎的主要路径。相较于粉粒、黏粒含量较高的土壤团聚体,砂粒含量较高的土壤团聚体向1~0.5,0.5~0.25 mm粒径的转化贡献率整体偏低。基于REE示踪得到的>0.25 mm各粒径团聚体质量整体被低估,低估范围为-27.96%^-11.08%;而<0.25 mm团聚体质量则被高估,高估范围为3.65%~22.73%。基于各粒径土壤团聚体的REE量化值建立了校正关系,可将计算相对误差降低至0.04%~16.24%。