Expression pattern and functions of autophagy-related gene atg5 in zebrafish organogenesis

The implications of autophagy-related genes in serious neural degenerative diseases have been well documented. However, the functions and regulation of the family genes in embryonic development remain to be rigorously studied. Here, we report on for the first time the important role of atg5 gene in zebrafish neurogenesis and organogenesis as evidenced by the spatiotemporal expression pattern and functional analysis. Using morpholino oligo knockdown and mRNA overexpression, we demonstrated that zebrafish atg5 is required for normal morphogenesis of brain regionalization and body plan as well as for expression regulation of neural gene markers: gli1, huC, nkx2.2, pink1, β-synuclein, xb51 and zic1. We further demonstrated that ATG5 protein is involved in autophagy by LC3-II/LC3I ratio and rapamycin-induction experiments, and that ATG5 is capable of regulating expression of itself gene in the manner of a feedback inhibition loop. In addition, we found that expression of another autophagy-related gene, atg12, is maintained at a higher constant level like a housekeeping gene. This indicates that the formation of the ATG12–ATG5 conjugate may be dependent on ATG5 protein generation and its splicing, rather than on ATG12 protein in zebrafish. Importantly, in the present study, we provide a mechanistic insight into the regulation and functional roles of atg5 in development of zebrafish nervous system.     

Gene Ontology-based function prediction of long non-coding RNAs using bi-random walk

Background

With the development of sequencing technology, more and more long non-coding RNAs (lncRNAs) have been identified. Some lncRNAs have been confirmed that they play an important role in the process of development through the dosage compensation effect, epigenetic regulation, cell differentiation regulation and other aspects. However, the majority of the lncRNAs have not been functionally characterized. Explore the function of lncRNAs and the regulatory network has become a hot research topic currently.

Methods

In the work, a network-based model named BiRWLGO is developed. The ultimate goal is to predict the probable functions for lncRNAs at large scale. The new model starts with building a global network composed of three networks: lncRNA similarity network, lncRNA-protein association network and protein-protein interaction (PPI) network. After that, it utilizes bi-random walk algorithm to explore the similarities between lncRNAs and proteins. Finally, we can annotate an lncRNA with the Gene Ontology (GO) terms according to its neighboring proteins.

Results

We compare the performance of BiRWLGO with the state-of-the-art models on a manually annotated lncRNA benchmark with known GO terms. The experimental results assert that BiRWLGO outperforms other methods in terms of both maximum F-measure (F_max) and coverage.

Conclusions

BiRWLGO is a relatively efficient method to predict the functions of lncRNA. When protein interaction data is integrated, the predictive performance of BiRWLGO gains a great improvement.

     

病毒治癌

病毒治疗癌症近来有新进展．分子生物学技术的发展,大大促进了病毒在寄主专一性、基因传递的定位性及溶解癌细胞方面的研究：选择适当的病毒种类,根据癌细胞特征性表面蛋白的特性修饰病毒的表面蛋白,使之对癌细胞具有更强的亲和性;去除病毒中原有的致病基因如癌基因,并根据靶细胞的特性针对性地改造有关病毒基因如插入适当的抗癌基因,可能构建成安全有效的工程病毒用于癌症治疗．病毒治癌除具有较强的特异性外,和其他基因治疗方法相比,还具有能利用寄主细胞进行自我复制、级联放大以克服初始剂量不足等特点．     

牛α—S1—酪蛋白—乙肝病毒表面抗原融合基因在转基因羊中的表达

利用ＤＮＡ重组技术,将牛α－Ｓ１酪蛋白调控区基因（１６ｋｂ的片段）与乙肝表面抗原基因拼接,构建了融合基因表达构件：λ１０６。构件经转基因技术转入山羊受精卵。待受精卵发育至成熟个体并分娩、泌乳,ＥＬＩＳＡ检测其乳汁中的ＨＢｓＡｇ,证明所构建的牛α－Ｓ１酪蛋白基因表达构件是可以在羊乳腺中表达乙肝表面抗原基因,并将其表达产物分泌到乳汁中。     

Some aspects of the growth of Gracilaria tenuistipitata in pond culture

The effect of temperature, salinity, nitrogen, culture density and depth on the growth of Gracilaria tenuistipitata were investigated between April 1985 and March 1986 in outdoor ponds in Guangxi Province, South China. The mean annual growth rate was 2.4% per day. Under favourable temperatures of 20–30 °C, daily growth rate may reach as high as 3.3%. Salinity had an obvious effect on growth and photosynthesis and growth peaked at 21, with a broad plateau between 7–27. Growth experiments showed that a total nitrogen (NH₄-N plus NO₃-N) concentration of 4 M was sufficient to enable the plants to maintain a daily growth rate of 2.7%. The best growth of the plant was obtained at a culture density of 0.5–1 kg m^–2 and a culture depth of 30 cm in the pond.     

Association of Serum Calcium and Hypertension Among Adolescents Aged 12–17 Years in the Rural Area of Northeast China

Calcium plays an important role in regulating body homeostasis. Several studies have reported the association between serum calcium and cardiovascular disease in adults. However, studies assessing the relationship between serum calcium and hypertension were limited, especially in subject populations of adolescents. The aim of the present study was to examine the association of serum calcium levels and blood pressure levels among adolescents in the rural area of Northeast China. A total of 2,023 students participated in this study, including 894 boys and 1,129 girls, aged from 12 to 17 years old. We measured the body weight, height, systolic blood pressure (SBP), diastolic blood pressure (DBP), and serum calcium concentrations of all eligible subjects, and the body mass index (BMI) was calculated from body weight and height. Childhood hypertension was defined as SBP and/or DBP ≥95th percentile for age and gender. According to the results of multivariable linear and logistic regression analysis, we found that higher serum calcium levels were positively associated with childhood hypertension. In comparison with serum calcium levels ≤2.37 mmol/L, the multivariable odds ratio (95 % confidence interval) of hypertension among adolescents with serum calcium levels ≥2.53 mmol/L was 1.89 (1.41–2.53; P trend?<?0.001). In addition, higher serum calcium levels were also positively associated with average difference in SBP and DBP; the average differences (95 % confidence interval) were 4.22 (2.74–5.83; P trend?<?0.001) and 2.23(1.00–3.46; P trend?<?0.001), respectively. In conclusion, higher serum calcium concentrations were found to have an association with higher blood pressure levels and higher prevalence of hypertension in the young population.     

Phosphorylation of Bcl-associated death protein (Bad) by erythropoietin-activated c-Jun N-terminal protein kinase 1 contributes to survival of erythropoietin-dependent cells

The glycoprotein erythropoietin (Epo) is a hematopoietic cytokine necessary for the survival of erythrocytes from immature erythroid cells. The mitogen-activated c-Jun N-terminal kinase 1 (JNK1) plays an important role in the proliferation and survival of erythroid cells in response to Epo. However, the precise mechanism of JNK1 activation promoting erythroid cell survival is incompletely understood. Here, we reported that JNK1 is required for Epo-mediated cell survival through phosphorylation and inactivation of the pro-apoptotic, Bcl-2 homology domain 3 (BH3)-only Bcl-associated death protein (Bad). Upon Epo withdrawal, HCD57 cells, a murine Epo-dependent cell line, displayed increased apoptotic cell death that was associated with decreased JNK1 activity. Epo withdrawal-induced apoptosis was promoted by inhibition of JNK1 activity but suppressed by expression of a constitutively active JNK1. Furthermore, Epo-activated JNK1 phosphorylated Bad at threonine 201, thereby inhibiting the association of Bad with the anti-apoptotic molecule B-cell lymphoma-extra large (Bcl-X(L)). Replacement of threonine 201 by alanine in Bad promoted Epo withdrawal-induced apoptosis. Thus, our results provide a molecular mechanism by which JNK1 contributes to the survival of erythroid cells.     

Co-expression of multiple gene constructs in transgenic mice

Multiple-transgene co-integration offers a powerful means by which several transgenes can be co-expressed in mammary glands. Independent gene constructs, including bovine -casein-hG-CSF, mWAP-hEPO, and CMV-EGFP, were co-injected into fertilized mouse eggs whereupon 32% (17/54) of the transgenic mice showed integration of all the three constructs. The co-expression ratio of hG-CSF and hEPO proteins in the mouse milk was up to 54% (6/11), attributable to co-integration. Maximal expression of human EPO and G-CSF was about 1 mg l^–1 and 540 mg l^–1 milk, respectively. There was an inverse relationship between transgene fragment length and integration ratio, and evidence that co-integration events are favoured above single integration events, suggesting that integration of multiple genes may be more facilitated than a single gene. The results have important practical implications for the generation of mammary gland bioreactors, multiple transgene co-integration appearing to be a useful strategy for generating animals expressing several transgenes simultaneously.     

miR-29b and miR-29c Are Involved in Toll-Like Receptor Control of Glucocorticoid-Induced Apoptosis in Human Plasmacytoid Dendritic Cells

Glucocorticoids (GCs) are frequently used to treat many of the acute disease manifestations associated with inflammatory and autoimmune disorders. However, Toll-like receptor (TLR) pathway-activated plasmacytoid dendritic cells (pDCs) are resistant to GC-induced apoptosis, which leads to the inefficiency of GCs in the treatment of type I interferon-related autoimmune diseases, such as systemic lupus erythematosus (SLE). Therefore, compounds promoting pDC apoptosis may be helpful for improving the efficacy of GCs. In this study, we performed screening to identify microRNAs (miRNAs) involved in TLR-inhibited GC-induced pDC apoptosis and found an array of miRNAs that may regulate pDC apoptosis. Among those demonstrating altered expression, 6 miRNAs were inhibited in TLR-activated pDCs. Bioinformatics analysis and functional studies indicated that miR-29b and miR-29c were 2 key miRNAs involved in TLR-inhibited GC-induced pDC apoptosis. Furthermore, both of these miRNAs promoted pDC apoptosis by directly targeting Mcl-1 and Bcl-2 in human primary pDCs. Our findings provide new targets that could improve the efficacy of GCs for the treatment of SLE.