Differentiation of immortalized epithelial cells derived from cystic fibrosis airway submucosal glands

Summary Cystic fibrosis (CF) involves abnormalities in mucus production and secretion of the airway. Studies of the regulation of airway mucin production and secretion has been difficult due to the lack of in vitro models of the airway epithelial cells which express functional differentiation. Because the majority of the mucin in the airway is apparently produced by the submucosal glands, we have focused our attention on the development of cell culture models of human airway submucosal glands. This report describes the propagation of CF airway submucosal gland epithelial cells which continue to express mucin production. The CF bronchus was obtained from a 31-yr-old patient who received a double lung transplant. The glands were dissected out and primary cultures prepared by the explant/outgrowth procedure. The cells were immortalized by infection with Adl2-SV40 hybrid virus. The cultures are maintained in serum-free keratinocyte basal medium supplemented with insulin (5μg/ml), hydrocortisone (0.5μg/ml), epidermal growth factor (10 ng/ml), bovine pituitary extract (25μg/ml), and antibiotics. Cultures were passaged using 0.125% trypsin in Ca⁺² and Mg⁺²-free Hanks’, balanced salt solution. Polymerase chain reaction (PCR) analysis demonstrated that the cells were homozygous for the ΔF508 mutation. Morphologic observations showed that the cells were epithelial and were interconnected by sparsely distributed desmosomes. Their cytoplasm contained secretory-type structures including abundant Golgi, rough endoplasmic reticulum, and secretory vesicles. Immunofluorescent studies determined that all cells were positive for cytokeratins, mucin glycoconjugates, and cystic fibrosis transmembrane conductance regulator. The cultures secreted substantial amounts of mucin glycoproteins and expressed the MUC-2 mucin gene. Patch clamp experiments revealed that the cells expressed defective Cl⁻ channels which were not activated by Forskolin.     

Ar^＋,远红外激光,γ射线单一及复合处理水稻的诱变效果

用Ar~+、远红外两种激光和~(60)Co—r射线单一或复合处理两个籼稻品种的干种子,分析和比较了不同处理对水稻的当代生物学效应及处理二代的变异频率。结果表明,两种激光对当代的几个性状均表现为刺激效应,并有减轻,射线辐射损伤的作用,复合处理二代的变异频率和变异类型数明显高于相应的单一处理。说明复合处理是提高激光育种效果的有效方法。     

Release of Prostaglandin E2 by Human Mononuclear Cells Exposed to Heat-Killed Salmonella typhi

Human mononuclear cells pre-labeled with [³H]arachidonic acid were shown to release metabolites following in vitro addition of heat-killed Salmonella typhi (HKST). The amount of label released was significantly higher than that seen with live S. typhi (LST). Addition of increasing amounts of HKST resulted in an increased release of metabolites. Enzyme immunoassay of the culture supernatants revealed that the bulk of the metabolite released was prostaglandin E₂ (PGE₂). Leukotriene B₄ (LTB₄) and leukotriene C₄ (LTC₄) were not detectable in the culture supernatants. The significance and implications of these results are discussed.     

Avicularin alleviates acute liver failure by regulation of the TLR4/MyD88/NF-κB and Nrf2/HO-1/GPX4 pathways to reduce inflammation and ferroptosis

Acute liver failure (ALF) is an inflammation-mediated hepatocyte death process associated with ferroptosis. Avicularin (AL), a Chinese herbal medicine, exerts anti-inflammatory and antioxidative effects. However, the protective effect of AL and the mechanism on ALF have not been reported. Our in vivo results suggest that AL significantly alleviated lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced hepatic pathological injury, liver enzymes, inflammatory cytokines, reactive oxygen species and iron levels and increased the antioxidant enzyme activities (malondialdehyde and glutathione). Our further in vitro experiments demonstrated that AL suppressed inflammatory response in LPS-stimulated RAW 264.7 cells via blocking the toll-like receptor 4 (TLR4)/myeloid differentiation protein-88 (MyD88)/nuclear factor kappa B (NF-κB) pathway. Moreover, AL attenuated ferroptosis in D-GalN-induced HepG2 cells by activating the nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1)/glutathione peroxidase 4 (GPX4) pathway. Therefore, AL can alleviate inflammatory response and ferroptosis in LPS/D-GalN-induced ALF, and its protective effects are associated with blocking TLR4/MyD88/NF-κB pathway and activating Nrf2/HO-1/GPX4 pathway. Moreover, AL is a promising therapeutic option for ALF and should be clinically explored.     

GPD1L inhibits renal cell carcinoma progression by regulating PINK1/Parkin-mediated mitophagy

Few approaches have been conducted in the treatment of renal cell carcinoma (RCC) after nephrectomy, resulting in a high mortality rate in urological tumours. Mitophagy is a mechanism of mitochondrial quality control that enables selective degradation of damaged and unnecessary mitochondria. Previous studies have found that glycerol-3-phosphate dehydrogenase 1-like (GPD1L) is associated with the progression of tumours such as lung cancer, colorectal cancer and oropharyngeal cancer, but the potential mechanism in RCC is still unclear. In this study, microarrays from tumour databases were analysed. The expression of GPD1L was confirmed by RT–qPCR and western blotting. The effect and mechanism of GPD1L were explored using cell counting kit 8, wound healing, invasion, flow cytometry and mitophagy-related experiments. The role of GPD1L was further confirmed in vivo. The results showed that GPD1L expression was downregulated and positively correlated with prognosis in RCC. Functional experiments revealed that GPD1L prevented proliferation, migration and invasion while promoting apoptosis and mitochondrial injury in vitro. The mechanistic results indicated that GPD1L interacted with PINK1, promoting PINK1/Parkin-mediated mitophagy. However, inhibition of PINK1 reversed GPD1L-mediated mitochondrial injury and mitophagy. Moreover, GPD1L prevented tumour growth and promoted mitophagy by activating the PINK1/Parkin pathway in vivo. Our study shows that GPD1L has a positive correlation with the prognosis of RCC. The potential mechanism involves interacting with PINK1 and regulating the PINK1/Parkin pathway. In conclusion, these results reveal that GPD1L can act as a biomarker and target for RCC diagnosis and therapy.     

Al-(L-丙氨酸)胰岛素晶体学研究

应用微量过饱和静置法在含柠檬酸钠和氯化钠的晶体生长体系中得到适合Ｘ射线衍射分析用的Ａｌ－（Ｌ－丙氯酸）胰岛素单晶体。晶体属于三方晶系,空间群为Ｒ３晶体衍射分辨率可达１．８Ａ以上。晶胞参数：ａＨ＝８０．８９Ａ,ｂＨ＝８０．８９Ａ,ｃＨ＝３７＼６４Ａ,α＝β＝９０°,γ＝１２０°。每个结晶学不对称单位含有两个Ａｌ－（Ｌ－丙氨酸）胰岛素分子。     

Multikink quantile regression for longitudinal data with application to progesterone data analysis

Motivated by investigating the relationship between progesterone and the days in a menstrual cycle in a longitudinal study, we propose a multikink quantile regression model for longitudinal data analysis. It relaxes the linearity condition and assumes different regression forms in different regions of the domain of the threshold covariate. In this paper, we first propose a multikink quantile regression for longitudinal data. Two estimation procedures are proposed to estimate the regression coefficients and the kink points locations: one is a computationally efficient profile estimator under the working independence framework while the other one considers the within-subject correlations by using the unbiased generalized estimation equation approach. The selection consistency of the number of kink points and the asymptotic normality of two proposed estimators are established. Second, we construct a rank score test based on partial subgradients for the existence of the kink effect in longitudinal studies. Both the null distribution and the local alternative distribution of the test statistic have been derived. Simulation studies show that the proposed methods have excellent finite sample performance. In the application to the longitudinal progesterone data, we identify two kink points in the progesterone curves over different quantiles and observe that the progesterone level remains stable before the day of ovulation, then increases quickly in 5 to 6 days after ovulation and then changes to stable again or drops slightly.     

Effects of Allelochemicals on Root Growth and Pod Yield in Response to Continuous Cropping Obstacle of Peanut

Continuous cropping (CC) obstacle is a major threat in legume crops production; however, the underlying mechanisms concerning the roles allelochemicals play in CC obstacle are poorly understood. The current 2-year study was conducted to investigate the effects of different kinds and concentrations of allelochemicals, p-hydroxybenzoic acid (H), cinnamic acid (C), phthalic acid (P), and their mixtures (M) on peanut root growth and productivity in response to CC obstacle. Treatment with H, C, P, and M significantly decreased the plant height, dry weight of the leaves and stems, number of branches, and length of the lateral stem compared with control. Exogenous application of H, C, P, and M inhibited the peanut root growth as indicated by the decreased root morphological characters. The allelochemicals also induced the cell membrane oxidation even though the antioxidant enzymes activities were significantly increased in peanut roots. Meanwhile, treatment with H, C, P, and M reduced the contents of total soluble sugar and total soluble protein. Analysis of ATPase activity, nitrate reductase activity, and root system activity revealed that the inhibition effects of allelochemicals on peanut roots might be due to the decrease in activities of ATPase and NR, and the inhibition of root system. Consequently, allelochemicals significantly decreased the pod yield of peanut compared with control. Our results demonstrate that allelochemicals play a dominant role in CC obstacle-induced peanut growth inhibition and yield reduction through damaging the root antioxidant system, unbalancing the osmolytes accumulation, and decreasing the activities of root-related enzymes.

     

烟草品种、育苗方式和烟蚜来源对烟蚜茧蜂寄生能力的影响

【目的】本研究旨在筛选出适合烟蚜茧蜂Aphidius gifuensis寄生烟蚜Myzus persicae的烟草品种、育苗方式和烟蚜来源,为烟蚜茧蜂人工繁殖和复壮提供科学参考。【方法】采用盆栽法,调查了不同烟草品种(云烟87、K326、翠碧1号和红花大金元)繁殖烟蚜若蚜、不同来源烟蚜(来自烟区、来自非烟区和室内繁殖种群)若蚜和烟草不同育苗方式(土壤育苗和湿润育苗)繁殖烟蚜若蚜时,烟蚜茧蜂成蜂对烟蚜若蚜的寄生率以及成蜂羽化率、寿命、体型大小和雌蜂比。【结果】结果表明,不同烟草品种繁殖烟蚜若蚜对烟蚜茧蜂成蜂寄生率存在显著影响。其中,在红花大金元和翠碧1号繁殖烟蚜上的烟蚜茧蜂成蜂寄生率分别比在K326繁殖烟蚜上的烟蚜茧蜂成蜂寄生率高19.00%和14.00%,但对烟蚜茧蜂的其他指标没有显著影响;不同烟蚜来源对烟蚜茧蜂寄生率也存在显著影响,其中,烟蚜茧蜂成蜂对非烟区烟蚜若蚜和烟区烟蚜若蚜的寄生率比对室内繁殖种群烟蚜若蚜的寄生率分别高20.25%和16.75%,但对烟蚜茧蜂成蜂的羽化率、寿命、体型大小和雌蜂比没有显著影响;烟草不同育苗方式对烟蚜茧蜂成蜂的寄生率无显著影响。【结论】烟蚜茧蜂对烟蚜...     

中国旧石器原料开发策略的研究进展

石料开发与获取是石器生产操作链的重要环节,也是史前人地关系研究的重要内容,能够反映史前人类的资源开发策略、环境认知与适应能力、移动模式和交流贸易网络等。相比于旧大陆西侧,东亚地区的燧石和黑曜岩等优质石料产地相对稀缺,中国许多旧石器考古遗址的石器原料主要来自遗址周边质量欠佳的脉石英、石英岩和一般燧石等,而相关的旧石器时代石料开发策略系统研究也相对较少。本文对中国目前已开展的旧石器时代石料开发策略研究进行系统梳理分析和总结发现,从旧石器时代早期到晚期末段,中国境内史前人类的石料开发策略存在明显的时空变化,主要体现在石料类型组成、产地选取和开发方式三个方面,可能主要受到史前人类流动性变化、石器技术演变和环境变化等因素的共同影响。综合而言,开展旧石器时代遗址的石料开发策略研究,对于理解中国境内史前人类的行为模式和交流迁徙等有重要意义。