Nitric oxide in the pathogenesis of diffuse pulmonary fibrosis

By studying the responses of nitric oxide in pulmonary fibrosis, the role of inducible nitric oxide synthase in diffuse pulmonary fibrosis as caused by lipopolysaccharide (LPS) treatment was investigated. When compared to rats treated with LPS only, the rats pretreated with 1400W (an iNOS-specific inhibitor) were found to exhibit a reduced level in: (i) NOx (nitrate/nitrite) production, (ii) collagen type I protein expression, (iv) soluble collagen production, and (iv) the loss of body weight and carotid artery PO2. In the pulmonary fibroblast culture, exogenous NO from LPS-stimulated secretion by macrophages or from a NO donor, such as DETA NONOate, was observed to induce the expression of TIMP-1, HSP47, TGF-beta1, and collagen type I as well as the phosphorylation of SMAD-2. After inhalation of NO for 24 h, an up-regulation of collagen type I protein was also noted to occur in rat pulmonary tissue. The results suggest that the NO signal pathway enhanced the expression of TGF-beta1, TIMP-1, and HSP47 in pulmonary fibroblasts, which collectively demonstrate that the NO signal pathway could activate the SMAD-signal cascade, by initiating a rapid increase in TGF-beta1, thereby increasing the expression of TIMP-1 and HSP47 in pulmonary fibroblasts, and play an important role in pulmonary fibrosis.     

TMEM41B and VMP1 modulate cellular lipid and energy metabolism for facilitating dengue virus infection

Transmembrane Protein 41B (TMEM41B) and Vacuole Membrane Protein 1 (VMP1) are two ER-associated lipid scramblases that play a role in autophagosome formation and cellular lipid metabolism. TMEM41B is also a recently validated host factor required by flaviviruses and coronaviruses. However, the exact underlying mechanism of TMEM41B in promoting viral infections remains an open question. Here, we validated that both TMEM41B and VMP1 are essential host dependency factors for all four serotypes of dengue virus (DENV) and human coronavirus OC43 (HCoV-OC43), but not chikungunya virus (CHIKV). While HCoV-OC43 failed to replicate entirely in both TMEM41B- and VMP1-deficient cells, we detected diminished levels of DENV infections in these cell lines, which were accompanied by upregulation of the innate immune dsRNA sensors, RIG-I and MDA5. Nonetheless, this upregulation did not correspondingly induce the downstream effector TBK1 activation and Interferon-beta expression. Despite low levels of DENV replication, classical DENV replication organelles were undetectable in the infected TMEM41B-deficient cells, suggesting that the upregulation of the dsRNA sensors is likely a consequence of aberrant viral replication rather than a causal factor for reduced DENV infection. Intriguingly, we uncovered that the inhibitory effect of TMEM41B deficiency on DENV replication, but not HCoV-OC43, can be partially reversed using exogenous fatty acid supplements. In contrast, VMP1 deficiency cannot be rescued using the metabolite treatment. In line with the observed phenotypes, we found that both TMEM41B- and VMP1-deficient cells harbor higher levels of compromised mitochondria, especially in VMP1 deficiency which results in severe dysregulations of mitochondrial beta-oxidation. Using a metabolomic profiling approach, we revealed distinctive global dysregulations of the cellular metabolome, particularly lipidome, in TMEM41B- and VMP1-deficient cells. Our findings highlight a central role for TMEM41B and VMP1 in modulating multiple cellular pathways, including lipid mobilization, mitochondrial beta-oxidation, and global metabolic regulations, to facilitate the replication of flaviviruses and coronaviruses.     

烟草初生胚乳细胞在微室培养中的发育

烟草初生胚乳细胞在微室培养中的发育 李师弢 房克凤 杨弘远*     

Construction and Analysis of cDNA Libraries from Proembryos and just Differentiating Young Embryos in Rice

Two cDNA libraries were constructed from microdissected 214 rice proembryos (2-3 d after pollination) and 121 just differentiating young embryos (3-5 d after pollination) respectively through RT-PCR technique. The primary libraries had a total of 3.7×10 phages for the proembryos and a total of 2.5×10 phages for the just differentiating young embryos, in which 96% of the phages were recombinants. Insert sizes ranging from 400 bp to 3?500 bp were obtained. All of theabove mentioned accorded with the general requirements of cDNA library construction.     

深黄被孢霉中微生物油脂的提取工艺

采用微波细胞破胞法处理深黄被孢霉,并对破胞后的菌体进行甲醇萃取油脂过程的研究。通过测量细胞内蛋白质释放量,确定微波处理的较佳条件为微波强度420 W,反应2 min。对微波后菌体进行扫描电镜观察,菌丝出现孔洞和裂纹,结构完全被破坏,达到微波破胞的效果;再直接进行甲酯化反应,通过正交试验考察固液比、反应温度、KOH浓度以及反应时间对甲酯化得率的影响。结果表明微生物油脂甲酯化的最佳条件:固液比(g/mL)1∶5,温度50℃,KOH-甲醇质量分数2.5%,时间2 h,此条件下干菌含油率为51.3%。     

Tankyrase 2 poly(ADP-ribose) polymerase domain-deleted mice exhibit growth defects but have normal telomere length and capping

Regulation of telomere length maintenance and capping are a critical cell functions in both normal and tumor cells. Tankyrase 2 (Tnks2) is a poly(ADP-ribose) polymerase (PARP) that has been shown to modify itself and TRF1, a telomere-binding protein. We show here by overexpression studies that tankyrase 2, like its closely related homolog tankyrase 1, can function as a positive regulator of telomere length in human cells, dependent on its catalytic PARP activity. To study the role of Tnks2 in vivo, we generated mice with the Tnks2 PARP domain deleted. These mice are viable and fertile but display a growth retardation phenotype. Telomere analysis by quantitative fluorescence in situ hybridization (FISH), flow-FISH, and restriction fragment analysis showed no change in telomere length or telomere capping in these mice. To determine the requirement for Tnks2 in long-term maintenance of telomeres, we generated embryonic stem cells with the Tnks2 PARP domain deleted and observed no change, even upon prolonged growth, in telomere length or telomere capping. Together, these results suggest that Tnks2 has a role in normal growth and development but is not essential for telomere length maintenance or telomere capping in mice.     

论草划原区和荒漠草原区在宁夏东部的界限

以盐池县为主的宁夏东部草原地区,地处典型草原向荒漠草原的过渡地带,饲养着全自治区近五分之二的二毛裘皮滩羊,是区内重要的畜牧业生产基地之一。这一地区在1955—1961年间曾由不同的专业队进行过多次自然资源综合考察[6,8,10,17],主要单位有中国科学院黄河中游水土保持综合考察队,中国科学院治沙工作队,中国科学院内蒙古宁夏综合考察队以及自治区综合勘察队和宁夏农学院等。虽然积累了一些有关这一地区草原植被方面的资料,但正式发表的不多,并且在草原界线的划分上由于认识的不同,分歧较大。如划分界线时有的主要考虑地形或土壤因子[8],而对植被本身的差异注意得不够,甚至建群针茅的种类也还没有搞清楚[17];还有的把隐域植被(沙生植被)与地带性植被混同起来,并过分强调少量入侵的荒漠植物(刺叶柄棘豆)的作用。1977年以来我们连续在这一地区进行草场植被资源调查和草原生态定位研究,现就我们的认识对宁夏东部草原的属性、界线及其资源特征,提出一些看法。     

大豆—根瘤菌共生固氮氢代谢的研究

检测了四株大豆根瘤菌在不同的大豆品种上形成根瘤的放氢、吸氢、固氮活性及豆血红蛋白的含量;同时测定了植株干物质的积累。结果表明,所有固氮根瘤都放氢,自生条件下Hup~-根瘤菌形成的根瘤仍不具吸氢活性,相对固氮率在0.75左右。而Hup~(?)菌株根瘤的相对固氮率在0.91～1之间。寄主植物对Hup~(?)菌株的吸氢活性有影响。相关分析表明,根瘤的豆血红蛋白与吸氢活性呈负相关。干物质积累与固氮酶活性关系最密切,氢酶活性的影响是次要的。     

Nitric oxide produced by iNOS is associated with collagen synthesis in keloid scar formation

Nitric oxide (NO) has emerged as an important mediator of many physiological functions. Recent reports have shown that NO participates in the wound healing process, however, its role in keloid formation remains unclear. This study aimed to investigate the effect of NO on keloid fibroblasts (KF) and to determine the levels of inducible nitric oxide synthase (iNOS) expression in clinical specimens of keloid. Scar tissue from seven keloid patients with matched perilesion skin tissue controls was studied for inducible nitric oxide synthase expression and location. In addition, primary keloid and normal scar skin fibroblast cultures were set up to investigate the effects of NO in inducing collagen type I expression. Inducible nitric oxide synthase expression, and NO production were elevated in keloid scar tissues but not in matched perilesion skin tissues. Furthermore, exposure of KF to exogenous NO resulted in increased expression of collagen type I in a dose-dependent manner. NO exposure also induced time-course dependent collagen I expression that peaked at 24h in KF. Taken together, these results indicate that excess collagen formations in keloid lesion may be attributed to iNOS overexpression.