A rice chloroplast transit peptide sequence does not alter the cytoplasmic localization of sheep serotonin N‐acetyltransferase expressed in transgenic rice plants

Ectopic overexpression of melatonin biosynthetic genes of animal origin has been used to generate melatonin‐rich transgenic plants to examine the functional roles of melatonin in plants. However, the subcellular localization of these proteins expressed in the transgenic plants remains unknown. We studied the localization of sheep (Ovis aries) serotonin N‐acetyltransferase (OaSNAT) and a translational fusion of a rice SNAT transit peptide to OaSNAT (TS:OaSNAT) in plants. Laser confocal microscopy analysis revealed that both OaSNAT and TS:OaSNAT proteins were localized to the cytoplasm even with the addition of the transit sequence to OaSNAT. Transgenic rice plants overexpressing the TS:OaSNAT fusion transgene exhibited high SNAT enzyme activity relative to untransformed wild‐type plants, but lower activity than transgenic rice plants expressing the wild‐type OaSNAT gene. Melatonin levels in both types of transgenic rice plant corresponded well with SNAT enzyme activity levels. The TS:OaSNAT transgenic lines exhibited increased seminal root growth relative to wild‐type plants, but less than in the OaSNAT transgenic lines, confirming that melatonin promotes root growth. Seed‐specific OaSNAT expression under the control of a rice prolamin promoter did not confer high levels of melatonin production in transgenic rice seeds compared with seeds from transgenic plants expressing OaSNAT under the control of the constitutive maize ubiquitin promoter.     

Short-course preoperative radiotherapy with immediate surgery versus long-course chemoradiation with delayed surgery in the treatment of rectal cancer: A systematic review and meta-analysis

BackgroundLong-course chemoradiotherapy (LCRT) with delayed surgery or short-course radiotherapy (SCRT) with immediate surgery is probably the most frequent regimen in the treatment of rectal cancer. Debate is still going on whether SCRT or LCRT is more effective. So we performed this meta-analysis to evaluate the safety and efficacy of SCRT with immediate surgery versus LCRT with delayed surgery for the management of rectal cancer.MethodsLiterature were searched from PubMed, Embase, Web of science, Cochrane Library up to May, 2014. Quality of the randomized controlled trials (RCTs) was evaluated according to the Cochrane's risk of bias tool of RCT. RevMan 5.3 was used for statistical analysis. Pooled risk ratio (RR) and 95% confidence interval (CI) were calculated. Subgroup analysis and sensitivity analysis were employed to explore heterogeneity.Results16 trials were included in the qualitative systematic review. 12 trials were included in meta-analyses. 4 of them were RCTs; other 8 were non-RCTs. Meta-analysis demonstrated that there were no significant differences in overall survival (OS), disease free survival (DFS), local recurrence rate (LRR), distant metastasis rate (DMR), sphincter preservation rate, R0 resection rate and late toxicity. Compared with SCRT, LCRT obviously increased pCR rate [RR = 0.15, 95%CI (0.08, 0.28), P = 0.003], while LCRT obviously increased the grade 3–4 acute toxicity [RR = 0.13, 95%CI (0.06, 0.28), P < 0.00001].ConclusionsSCRT with immediate surgery is as effective as LCRT with delayed surgery for treatment of rectal cancer in terms of OS, DFS, LRR, DMR, Sphincter preservation rate, R0 resection rate and late toxicity. Though LCRT increased pCR rate, LCRT also increased acute toxicity compared with SCRT. SCRT is a better choice in centers with a long waiting list or lack of medical resources.     

Economic evaluation comparing From Home To Operation same day admission and preoperative admission one day prior to the surgery process : a randomized,controlled trial of laparoscopic cholecystectomy

ABSTRACT

Objective: A novel preoperative procedure From Home To Operation (FHTO) seeks to combat increasing operation and infection rates. This is the first prospective randomized controlled trial (RCT) comparing the cost-effectiveness and cost-utility of FHTO and conventional ward procedures for standardized Laparoscopic Cholecystectomy (LC).

Research design and methods: During 12/2004–7/2005, 47 patients with symptomatic gallstones were randomized to receive LC in the FHTO (28 patients) or in a conventional manner (19 patients) in a Finnish hospital setting. The 15D quality of life tool was administered at the baseline and 1 month after.

Main outcome measures: A stochastic approach over a month interval for hospital costs, length of postoperative stay, infection rate and Quality-Adjusted Life Years (QALY) was employed.

Results: Baseline group characteristics were similar. The mean health care costs with FHTO (1695 EUR) were significantly lower (?p < 0.001) than in the conventional arm (2234 EUR). The number of patients discharged on the first postoperative day was 27 (96.4%) and 15 (78.9%) with two (7.1%) infections in the FHTO and four (21.1%) in the conventional arm. A difference in QALYs gained (0.0174; p = 0.030) favouring FHTO was observed. According to a cost-effectiveness acceptability curve, the probability of FHTO being cost-effective was 99%. The results were robust to probabilistic sensitivity analyses.

Conclusions: FHTO can introduce substantial cost savings and have a positive impact on both clinical measures and quality of life. Studies with larger numbers of patients are needed to assess whether conventional ward procedure can be a source of infections, which can be avoided with FHTO.

Clinical Trial Registry: ICJME-qualified registry of the Hospital District of Helsinki and Uusimaa (number 217849).     

Hypocrellin B in hepatocellular carcinoma cells: Subcellular localization and sonodynamic damage

Abstract

Purpose: To study subcellular localization of hypocrellin B in hepatocellular carcinoma cells, and hypocrellin B-mediated sonodynamic action-induced cell damage.

Materials and methods: After incubation with 2.5 μM of hypocrellin B, human hepatocellular carcinoma HepG2 cells were exposed to ultrasound waves for 8 sec at an intensity of 0.46 W/cm². Clonogenic survival of HepG2 cells was measured using a colony forming assay and light microscope. Ultrastructural morphology was observed using transmission electron microscope (TEM) and mitochondrial membrane potential (MMP) was assessed using confocal laser scanning microcope (CLSM) after rhodamine 123 staining. Additionally, subcellular localization of hypocrellin B in HepG2 cells with organelle probe staining was also observed using CLSM.

Results: The colony forming units of HepG2 cells decreased substantially after sonodynamic treatment. The results of TEM showed microvilli disappearance, apoptotic body formation, swollen mitochondria with loss of cristae and mitochondrial myelin-like features (or membrane whorls). Collapse of MMP was found in the treated cells. Hypocrellin B was distributed in mitochondria and lysosomes as well as in endoplasmic reticulum and Golgi apparatus.

Conclusions: The findings demonstrated that sonodynamic action of hypocrellin B induced mitochondrial damage, survival inhibition, and apoptosis of HepG2 cells. Additionally, other subcellular organelles such as endoplasmic reticulum, Golgi apparatus and lysosomes were also the targets of hypocrellin B-mediated sonodynamic action as well as mitochondria.     

虚拟现实术前计划在特发性面肌痉挛微血管减压术中的应用

目的评价虚拟现实术前计划用于特发性面肌痉挛导微血管减压术的效果。方法收集33例做虚拟现实术前计划（计划组）和27例未做虚拟现实术前计划（对照组）的特发性面肌痉挛患者的病例资料进行回顾性分析。计划组患者分别行脑血管CT和头颅MRI检查，影像数据输入Vitrea虚拟现实系统进行三维重建，模拟枕下乙状窦后入路显露面听神经复合体，判断责任血管和面神经的空间关系，在术前计划基础上进行面神经微血管减压术。比较2组手术疗效和并发症发生率。结果2组患者责任血管均为小脑前下动脉，虚拟现实术前计划中面神经与责任血管空间关系清晰显示，责任血管判定结果与术中观察结果一致。计划组和对照组Cohen分级0级比例差异有统计学意义[100．0％（33例）比85．2％（23例）]（P=0．036），手术疗效总体良好率差异无统计学意义[100．0％（33例）比85．2％（23例）]（P=0．450）。并发症总体发生率差异无统计学意义[0．0％比11．1％（3例）]（P=0．085）。结论虚拟现实术前计划有助于清晰直观地显示面神经与责任血管的空间关系，可提高手术疗效和安全性。     

肿瘤抑制因子PTEN亚细胞定位与缺血-缺氧性脑损伤

第10号染色体缺失的磷酸酶和张力蛋白同源等位基因(PTEN)具有脂质和蛋白双重磷酸酶活性。脑组织缺血-缺氧后PTEN发生活化并向线粒体或胞核内转移,而这种在神经元内的不同亚细胞定位可能最终决定细胞命运。因此,阐明PTEN在脑缺血-缺氧后的亚细胞定位机制,将为各种急慢性神经退行性变疾病的靶向治疗和药物研发提供新思路。