Cationic micelle: A promising nanocarrier for gene delivery with high transfection efficiency

Micelles have demonstrated an excellent ability to deliver several different types of therapeutic agents, including chemotherapy drugs, proteins, small‐interfering RNA and DNA, into tumor cells. Cationic micelles, comprising self‐assemblies of amphiphilic cationic polymers, have exhibited tremendous promise with respect to the delivery of therapy genes and gene transfection. To date, research in the field has focused on achieving an enhanced stability of the micellar assembly, prolonged circulation times and controlled release of the gene. This review focuses on the micelles as a nanosized carrier system for gene delivery, the system‐related modifications for cytoplasm release, stability and biocompatibility, and clinic trials. In accordance with the development of synthetic chemistry and self‐assembly technology, the structures and functionalities of micelles can be precisely controlled, and hence the synthetic micelles not only efficiently condense DNA, but also facilitate DNA endocytosis, endosomal escape, DNA uptake and nuclear transport, resulting in a comparable gene transfection of virus.     

马来酸噻吗洛尔眼液联合拉坦前列素眼液对原发性开角型青光眼患者眼压的影响

目的:探讨马来酸噻吗洛尔眼液联合拉坦前列素眼液对原发性开角型青光眼患者眼压的影响。方法:选取2015年1月-2016年5月在我院接受治疗的原发性开角型青光眼患者84例,其中给予马来酸噻吗洛尔眼液治疗的42例记为对照组,给予拉坦前列素眼液联合马来酸噻吗洛尔眼液治疗的42例记为观察组,两组均治疗6个月。对比两组患者治疗过程中的眼压变化情况,并对比两组患者的临床总有效率、药物依从性和并发症情况。结果:观察组治疗2、4、6个月后的眼压均显著低于对照组(P0.05),观察组患者治疗6个月后与治疗前的眼压差值大于对照组(P0.05)。两组患者治疗2、4、6个月后的眼压呈下降趋势,两两比较差异均有统计学意义(P0.05)。观察组的临床总有效率95.24%显著高于对照组的80.95%(P0.05)。两组患者在治疗过程中结膜充血、眼内异物感、眼睛疼痛、视力模糊、味觉异常以及总并发症发生率对比差异不显著(P0.05)。观察组患者的药物依从性比例显著低于对照组(P0.05)。结论:马来酸噻吗洛尔眼液联合拉坦前列素眼液治疗原发性开角型青光眼患者具有较好的临床疗效,可显著降低患者眼压,同时具有较好的安全性,但药物依从性较差。     

Evaluation of tissue specificity and expression strength of rice seed component gene promoters in transgenic rice

Using stable transgenic rice plants, the promoters of 15 genes expressed in rice seed were analysed for their spatial and temporal expression pattern and their potential to promote the expression of recombinant proteins in seeds. The 15 genes included 10 seed storage protein genes and five genes for enzymes involved in carbohydrate and nitrogen metabolism. The promoters for the glutelins and the 13 kDa and 16 kDa prolamins directed endosperm-specific expression, especially in the outer portion (peripheral region) of the endosperm, whilst the embryo globulin and 18 kDa oleosin promoters directed expression in the embryo and aleurone layer. Fusion of the GUS gene to the 26 kDa globulin promoter resulted in expression in the inner starchy endosperm tissue. It should be noted that the 10 kDa prolamin gene was the only one tested that required both the 5' and 3' flanking regions for intrinsic endosperm-specific expression. The promoters from the pyruvate orthophosphate dikinase (PPDK) and ADP-glucose pyrophosphorylase (AGPase) small subunit genes were active not only in the seed, but also in the phloem of vegetative tissues. Within the seed, the expression from these two promoters differed in that the PPDK gene was only expressed in the endosperm, whereas the AGPase small subunit gene was expressed throughout the seed. The GUS reporter gene fused to the alanine aminotransferase (AlaAT) promoter was expressed in the inner portion of the starchy endosperm, whilst the starch branching enzyme (SBE1) and the glutamate synthase (GOGAT) genes were mainly expressed in the scutellum (between the endosperm and embryo). When promoter activities were examined during seed maturation, the glutelin GluB-4, 26 kDa globulin and 10 kDa and 16 kDa prolamin promoters exhibited much higher activities than the others. The seed promoters analysed here exhibited a wide variety of activities and expression patterns, thus providing many choices suitable for various applications in plant biotechnology.     

明目羊肝丸联合普拉洛芬滴眼液治疗围绝经期干眼症的临床研究

摘要 目的：探讨明目羊肝丸联合普拉洛芬滴眼液治疗围绝经期干眼症的临床疗效。方法：选取2019年7月至2020年8月在我院就诊的围绝经期干眼症患者103例206眼,根据随机数字表法将患者分为对照组（普拉洛芬滴眼液治疗）和实验组（对照组的基础上联合明目羊肝丸治疗）,分别为51例102眼和52例104眼,两组均治疗2个月。对比两组疗效、症状评分、泪液炎症因子、生存质量、Schirmer I试验（SIT）、泪膜破裂时间（BUT）、角膜荧光素染色（FL）评分及不良反应。结果：实验组治疗2个月后的临床总有效率优于对照组（P<0.05）。治疗2个月后,实验组眼干涩、视疲劳、异物感、灼热感症状评分、FL评分低于对照组（P<0.05）。治疗2个月后,实验组BUT、SIT高于对照组（P<0.05）。治疗2个月后,实验组泪液白介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）水平低于对照组（P<0.05）。治疗2个月后,实验组心理健康、眼痛、社会活动、一般健康评分高于对照组（P<0.05）。两组治疗过程中均无明显不良反应。结论：明目羊肝丸联合普拉洛芬滴眼液治疗围绝经期干眼症,可改善患者症状和泪膜稳定性,降低泪液炎症因子水平,提高患者生存质量,安全有效。     

视觉功能训练联合托吡卡胺滴眼液治疗假性近视的临床疗效观察

摘要 目的：探讨视觉功能训练联合托吡卡胺滴眼液在治疗假性近视中的临床疗效。方法：选取中国人民解放军陆军特色医学中心眼科和重庆佑佑宝贝妇儿医院眼科2019年2月至2019年8月收治的400例假性近视患儿作为研究对象,将其随机分为对照组和观察组(n=200)。对照组给予滴眼液缓解眼疲劳治疗,观察组则采用视觉功能训练联合托吡卡胺滴眼液治疗。比较两组患儿治疗前及治疗2个月、6个月后裸眼视力、眼轴长度、屈光度、正相对调节量和视疲劳评分的变化。结果：治疗2个月和6个月后,观察组患儿裸眼视力分别为(0.75±0.05)和(0.81±0.04),较对照组显著上升(P<0.05);屈光度分别为(+0.05±0.50)和(+0.06±0.48),明显高于对照组(P<0.05);正相对调节量检测结果依次是(-1.98±0.07)和(-2.51±0.24),较对照组明显增加(P<0.05)。两组患儿视觉感知双眼视功能均有明显改善,其中观察组视觉功能恢复显著高于对照组(P<0.05)。视疲劳调查显示观察组患儿视疲劳评分分别为(6.90±0.89)分和(3.91±0.89)分,显著低于对照组(P<0.05)。结论：视觉功能训练联合托吡卡胺滴眼液治疗能有效提高患儿裸眼视力、改善屈光度、正相对调节量和视觉感知功能,从而减少视疲劳,预防近视。     

芪明颗粒联合玻璃酸钠滴眼液治疗儿童中度干眼症的疗效及对泪液氧化应激和炎症因子的影响

摘要 目的：探讨芪明颗粒联合玻璃酸钠滴眼液治疗儿童中度干眼症的疗效及对泪液氧化应激和炎症因子的影响。方法：选取2019年8月~2021年12月期间来贵阳市妇幼保健院就诊的干眼症患儿,共计90例。入选的患儿根据随机数字表法分为对照组（玻璃酸钠滴眼液治疗）和观察组（芪明颗粒联合玻璃酸钠滴眼液治疗）,各为45例,两组均治疗3个月。对比两组疗效、临床指标、泪液氧化应激指标、炎症因子指标和不良反应。结果：观察组93.33%的临床总有效率较对照组68.89%高,差异有统计学意义（P<0.05）。两组不良反应发生率对比无差异（P>0.05）。观察组治疗3个月后泪膜破裂时间（BUT）、泪液分泌试验（SIT）较对照组高,角膜荧光染色（FL）评分较对照组低（P<0.05）。观察组治疗3个月后超氧化物歧化酶（SOD）高于对照组,丙二醛（MDA）、过氧化脂质（LPO）低于对照组（P<0.05）。观察组治疗3个月后白细胞介素1β（IL-1β）、白介素-6（IL-6）、超敏C反应蛋白（hs-CRP）、白介素-8（IL-8）低于对照组（P<0.05）。结论：儿童中度干眼症采用玻璃酸钠滴眼液联合芪明颗粒治疗,可有效降低患儿的氧化应激和炎症反应,且安全性较好。     

A model of drop size distribution for a system with evaporation

Abstract. The size and shape of drops on leaf surfaces strongly affect their persistence. The relationship between volume and exposed surface area of drops on wheat leaves and the log-normal drop size distribution in a wheat canopy after rain are used to derive equations to describe how the total volume and drop number change with evaporation. Firstly, the behaviour of a single drop as it evaporates is considered and then equations describing the change in a population of drops with an initial log-normal distribution are derived. The time taken for all the drops to reach complete dryness is about thirty times that for the same volume of water spread uniformly over the surface with the same potential evaporation rate.     

The effects of a stimulating intron on the expression of heterologous genes in Arabidopsis thaliana

Introns are often added to transgenes to increase expression, although the mechanism through which introns stimulate gene expression in plants and other eukaryotes remains mysterious. While introns vary in their effect on expression, it is unknown whether different genes respond similarly to the same stimulatory intron. Furthermore, the degree to which gene regulation is preserved when expression is increased by an intron has not been thoroughly investigated. To test the effects of the same intron on the expression of a range of genes, GUS translational fusions were constructed using the promoters of eight Arabidopsis genes whose expression was reported to be constitutive (GAE1, CNGC2 and ROP10), tissue specific (ADL1A, YAB3 and AtAMT2) or regulated by light (ULI3 and MSBP1). For each gene, a fusion containing the first intron from the UBQ10 gene was compared to fusions containing the gene's endogenous first intron (if the gene has one) or no intron. In every case, the UBQ10 intron increased expression relative to the intronless control, although the magnitude of the change and the level of expression varied. The UBQ10 intron also changed the expression patterns of the CNGC2 and YAB3 fusions to include strong activity in roots, indicating that tissue specificity was disrupted by this intron. In contrast, the regulation of the ULI3 and MSBP1 genes by light was preserved when their expression was stimulated by the intron. These findings have important implications for biotechnology applications in which a high level of transgene expression in only certain tissues is desired.     

Expression of inducible angiosperm promoters in a gymnosperm,Picea glauca (white spruce)

Electrical discharge particle acceleration was used to test the transient expression of numerous inducible angiosperm promoters in a gymnospermPicea glauca (white spruce). Promoter expression was assayed in three different tissues capable ofin vitro regeneration, zygotic embryos, seedlings and embryogenic callus. The promoters tested include the light-inducibleArabidopsis and soybean ribulose-1,5-bisphosphate small subunit promoters and a maize phosphoenolpyruvate carboxylase promoter; a soybean heat-shock-inducible promoter, a soybean auxin inducible promoter and a maize alcohol dehydrogenase promoter. Promoters were cloned into a promoter-less expression vector to form a promoter--glucuronidase-nopaline synthase 3 fusion. A similar construct was made using the cauliflower mosaic virus 35S (CaMV 35S) promoter as a control. All promoters were expressed in white spruce embryos, yet at levels lower than CaMV 35S. In addition, in the embryos the heat-shock and the alcohol dehydrogenase promoters showed inducible expression when given the proper induction stimulus. In seedlings, expression of all promoters was lower than in the embryos and expression was only inducible with the heat-shock promoter in the cotyledons. Of the tissues tested, the expression level of all promoters was lowest in embryogenic callus. Interestingly, the expression of the -glucuronidase gene in embryogenic callus was restricted to the proembryonal head cells regardless of the promoter used. These results clearly demonstrate the use of particle bombardment to test the transient expression of heterologous promoters in organized tissue and the expression of angiosperm promoters in a gymnosperm.     

复明片联合曲伏前列素滴眼液对原发性开角型青光眼患者眼部血流动力学和房水EPO、sCD44的影响

摘要 目的：探讨复明片联合曲伏前列素滴眼液对原发性开角型青光眼（POAG）患者眼部血流动力学和房水促红细胞生成素（EPO）、可溶性CD44（sCD44）的影响。方法：选择2017年4月~2020年11月在本院接受治疗的112例193眼的POAG患者,根据随机数字表法分为对照组（n=56,96眼）和研究组（n=56,97眼）,对照组患者接受曲伏前列素滴眼液治疗,研究组接受复明片联合曲伏前列素滴眼液治疗,对比两组疗效、眼部血流动力学和房水EPO、sCD44,视力、视野、眼压,观察治疗期间不良反应发生状况。结果：研究组的临床总有效率高于对照组（P<0.05）。治疗12周后,研究组平均光敏感度（MS）、视力高于对照组,24 h眼压平均值小于对照组（P<0.05）。治疗12周后,研究组舒张末期流速（EDV）、收缩期峰值流速（PSV）高于对照组,血流阻力系数（RI）小于对照组（P<0.05）。治疗12周后,研究组房水EPO、sCD44水平低于对照组（P<0.05）。两组不良反应发生率组间对比无统计学差异（P>0.05）。结论：复明片联合曲伏前列素滴眼液治疗POAG患者,可促进患者视力、视野、眼压改善,可能与调节眼部血流动力学和房水EPO、sCD44水平有关。     

Differential allelic expression of IL13 and CSF2 genes associated with asthma

An important area of genetic research is the identification of functional mechanisms in polymorphisms associated with diseases. A highly relevant functional mechanism is the influence of polymorphisms on gene expression levels (differential allelic expression, DAE). The coding single nucleotide polymorphisms (SNPs) CSF2^rs25882 and IL13^rs20541 have been associated with asthma. In this work, we investigated whether the mRNA expression levels of CSF2 or IL13 were correlated with these SNPs. Samples were analyzed by mass spectrometry-based quantification of gene expression. Both SNPs influenced gene expression levels (CSF2^rs25882: p_overall = 0.008 and p_{DAE samples} = 0.00006; IL13^rs20541: p_overall = 0.059 and p_{DAE samples} = 0.036). For CSF2, the expression level was increased by 27.4% (95% CI: 18.5%–35.4%) in samples with significant DAE in the presence of one copy of risk variant CSF2^rs25882-T. The average expression level of IL13 was increased by 29.8% (95% CI: 3.1%–63.4%) in samples with significant DAE in the presence of one copy of risk variant IL13^rs20541-A. Enhanced expression of CSF2 could stimulate macrophages and neutrophils during inflammation and may be related to the etiology of asthma. For IL-13, higher expression could enhance the functional activity of the asthma-associated isoform. Overall, the analysis of DAE provides an efficient approach for identifying possible functional mechanisms that link disease-associated variants with altered gene expression levels.     

Expression and regulation of genes associated with cell death during murine preimplantation embryo development

The newly fertilized preimplantation embryo depends entirely on maternal mRNAs and proteins deposited and stored in the oocyte prior to its ovulation. If the oocyte is not sufficiently equipped with maternally stored products, or if zygotic gene expression does not commence at the correct time, the embryo will die. One of the major abnormalities observed during early development is cellular fragmentation. We showed previously that cellular fragmentation in human embryos can be attributed to programmed cell death (PCD). Here, we demonstrate that the PCD that occurs during the 1-cell stage of mouse embryogenesis is likely to be regulated by many cell death genes either maternally inherited or transcribed from the embryonic genome. We have demonstrated for the first time the temporal expression patterns of nine cell death regulatory genes, and our preliminary experiments show that the expression of these genes is altered in embryos undergoing fragmentation. The expression of genes involved in cell death (MA-3, p53, Bad, and Bcl-xS) seems to be elevated, whereas the expression of genes involved in cell survival (Bcl-2) is reduced. We propose that PCD may occur by default in embryos that fail to execute essential developmental events during the first cell cycle. Mol. Reprod. Dev. 51:243–253, 1998. © 1998 Wiley-Liss, Inc.     

Expression profile of genes associated with mastitis in dairy cattle

In order to characterize the expression of genes associated with immune response mechanisms to mastitis, we quantified the relative expression of the IL-2, IL-4, IL-6, IL-8, IL-10, IFN-γ and TNF- α genes in milk cells of healthy cows and cows with clinical mastitis. Total RNA was extracted from milk cells of six Black and White Holstein (BW) cows and six Gyr cows, including three animals with and three without mastitis per breed. Gene expression was analyzed by real-time PCR. IL-10 gene expression was higher in the group of BW and Gyr cows with mastitis compared to animals free of infection from both breeds (p < 0.05). It was also higher in BW Holstein animals with clinical mastitis (p < 0.001), but it was not significant when Gyr cows with and without mastitis were compared (0.05 < p < 0.10). Among healthy cows, BW Holstein animals tended to present a higher expression of all genes studied, with a significant difference for the IL-2 and IFN- γ genes (p < 0.001). For animals with mastitis no significant difference in gene expression was observed between the two breeds. These findings suggest that animals with mastitis develop a preferentially cell-mediated immune response. Further studies including larger samples are necessary to better characterize the gene expression profile in cows with mastitis.