猪精浆蛋白基因PSP-Ⅰ和PSP-Ⅱ启动子序列克隆及生物信息学分析

分别用PCR方法扩增了1.7kbp和1.6kbp的猪PSP-Ⅰ和PSP-Ⅱ基因的启动子,并进行TA克隆,测序鉴定,测序结果用DNAstar程序与Genebank中的相应序列进行对比分析,结果显示与已发表序列的同源性分别为99.8%和96.3%。利用生物信息学的方法对克隆的1.7kbp和1.6kbp的猪PSP-Ⅰ和PSP-Ⅱ基因的启动子区进行了预测分析。PSP-Ⅰ和PSP-Ⅱ基因序列对比(mVista)分析发现,启动子0→-1000bp的保守性较高,其中0→-200bp核心启动子部分的序列同源性达到了100%,而-1000bp上游序列的保守性则较低。启动子的位置预测(Promoter Scan)结果显示,转录起始点上游200bp的区域为两基因的核心启动子位置。启动子区转录因子结合位点预测(TFSEARCH)发现,转录起始位点上游的1000bp区域内含有大量的顺式调控元件,并且得到了一系列潜在的转录因子结合位点。     

肝组织中NF-κB、TGF-β1及其Ⅰ型受体mRNA和HSC在肝纤维化中的改变及护肝片对其的影响

目的探讨中、晚期纤维化大鼠肝组织中肝星状细胞(HSC)的活化与增殖、核转录因子-κB(NF-κB)及转化生长因子-β1(TGF-β1)及其Ⅰ型受体(TβRⅠ)表达的改变及护肝片对其的影响。方法采用12.5%CCl4诱导的大鼠肝纤维化模型,自造模之日起,大鼠分组灌胃给药(护肝片921mg/kg)或溶媒,每日一次,直至8或13周末,分别处死动物,取左叶肝组织石蜡包埋,制作组织芯片。免疫组化S-P法检测大鼠肝组织α-平滑肌肌动蛋白(-αSMA)和NF-κB p65蛋白的表达,原位杂交检测TGF-β1及TβRⅠmRNA的表达;并用MetaMorph图像分析系统计数-αSMA阳性细胞数,对NF-κB p65蛋白、TGF-β1及TβRⅠmRNA的表达量进行定量分析。结果 1.模型复制8周和13周,模型组的肝损伤及其纤维化分级均明显高于正常组(P<0.01),护肝片组的肝损伤及其纤维化分级均轻于模型组。2.模型复制8周和13周,模型组活化的HSC(即-αSMA阳性细胞)数量较正常组明显增多,NF-κB p65蛋白、TGF-β1及TβRⅠmRNA的表达均较正常组明显增强(P<0.01);3.护肝片显著抑制8、13周纤维化肝组织HSC的活化与增殖和NF-κB p65蛋白、TGF-β1及TβRⅠmRNA的表达(P<0.01)。结论抑制HSC的活化与增殖和NF-κB p65蛋白与TGF-β1及TβRⅠmRNA的表达可能是护肝片抗肝纤维化作用的靶点之一。     

6种水蛭的COⅠ、12S rRNA和16S rRNA基因及分子进化分析

水蛭是一种常见的传统中药,为了解常见蛭类细胞色素氧化酶亚基Ⅰ（COⅠ）、12S rRNA和16S rRNA基因特征和水蛭分子系统进化关系。对常用的入药品种日本医蛭（Hirudo nipponia）、宽体金线蛭（Whitmania pigra）、尖细金线蛭（Whitmania acranulate）和相近物种菲牛蛭（Poecilobdella manillensis）、光润金线蛭（Whitmania laevis）及八目石蛭（Erpobdella octoculata）的COⅠ、12S rRNA和16S rRNA基因进行扩增、测序,利用Mega 5.0分析基因特征、颠换率、分化年代,利用PAUP＊4.10b和MrBayes 3.1.2构建分子系统树。结果表明6种水蛭的COⅠ、12S rRNA和16S rRNA基因全长分别为1534~1536 bp、709~744 bp、1129~1173 bp,GC含量分别为32.35%~34.79%、24.42%~28.49%、24.82%~27.02%,总体颠换率为0.002%~0.760%,分化年代为3.55×106a~9.85×106a;每种水蛭为单系群的支持值均≥82。说明COⅠ、12S rRNA和16S rRNA基因具有种间特异性,可用于6种水蛭的分类鉴别。     

Identification,regulation and physiological functions of multiple NADPH dehydrogenase complexes in cyanobacteria

Cyanobacteria possess multiple,functionally distinct NADPH dehydrogenase (NDH-1) complexes.In this mini-review,we describe the cyanobacterial NDH-1 complexes by focusing on their identification,regulatory properties,and multiple functions.The multiple functions can be divided into basic and extending functions,and the basic functions are compared with those in chloroplasts.Many questions related to cyanobacterial NDH-1 complexes remain unanswered and are briefly summarized here.     

Correlation between serum renin-angiotensin system (RAS) level and depression and anxiety symptoms in patients with Parkinson's disease

To investigate the correlation between serum renin-angiotensin system (RAS) level and Symptoms of anxiety and depression in Parkinson disease patients (PD). A number of 90 PD patients (47 males and 43 females) were collected on an empty stomach 12 h after stopping taking anti-PD medicines. ELISA has been found in Serum RAS ((Ang) I, Ang II, Ang (1–7), Angiotensin converting enzyme (ACE), ACE2). Depression scale (HAMD) and Anxiety scale (HAMA) in Hamilton are used for the assessment of signs of depression and anxiety. The 90 patients were diagnosed with moderate depression (HAMD score 8 ~ 19); in 32 of those (35.56 percent), and 12 (13.33%) were diagnosed as moderate and severe depression (HAMD score ≥ 20). 20 cases (22.22%) were diagnosed as possible anxiety disorder (HAMA score 7 ~ 13) and 16 cases (17.78%) as definite anxiety disorder (HAMA score ≥ 14). The association of serum Ang I, Ang II and Ang (1–7) with HAMD (r= − 0.820, P < 0.001; r = −0.846, P < 0.001) showed negative linkage with HAMD (r = −0.887, P < 0.003; P < 0.001; Negative correlation of the settings with HAMA (r = −0.850, P < 0.001; r = −0.887, P < 0.001; r = 0.003; r = 0.001, P < 0.001, Fig. 2, Fig. 3); The HAMD score and the HAMA score (all P > 0.05) were not associated to the serum ACE and ACE2. The serum Ang I, Ang II, and Ang (1–7) were found to be adversely associated with HAMD score (r = 0.826, P < 0,001; r = −0.818, p> >0,001; r = −0.876, P < 0,001; P = 0,001) P < 0,001; And have been negatively correlated (r = 0.870, Fig. 1, Fig. 2, Fig. 3) with AMA-scores (r = −0.876, P < 0.001, Table 1, Fig. 3), R = −0.862, P > 0.001; The HAMD score and the HAMA score (all P > 0.05) were not correlated to the serum ACE and ACE2. Finally, in PD patients, non-engine signs, including depression and anxiety, are normal. Thus, Serum levels Ang I, Ang II and Ang (1–7) were substantially decreased in female and male patients and associated with symptoms of depression and anxiety, ACE and ACE2 levels have not been attributed to signs of depression and anxiety. Serum Ang I, Ang II, and Ang (1–7) are important markers of depression and anxiety prevention and diagnosis in patients with DP.Table 1Comparison of serum ACE, ACE2, Ang I, Ang II, Ang (1–7) levels and HAMD and HAMA scores between male and female patients with PD.

Mechanism of the salt activation of laccase Lac15

Laccases (benzenediol: oxygen oxidoreductases, EC1.10.3.2) can oxidize various substrates, and those which are tolerant to and even activated by salts have attracted a lot of attention due to their application potential in certain industries. The mechanism of the salt activation of laccases is awaiting to be elucidated yet. Our previous study (Li, Xie et al. 2018) supposed that the salt activation of marine laccase Lac15 might be attributed to Cl^- ion specifically binding to some local sites to interfere substrate binding and/or electron transfer. In this study, we found two sites whose mutations resulted in elimination of the salt activation of Lac15’s activity towards catechol and dopamine respectively, and revealed that the mutations affected the activity by altering both E_m and k_cat, demonstrating the supposed mechanism. A model for the salt activation of laccases was accordingly proposed, albeit some details are to be elucidated.