DNA end resection is needed for the repair of complex lesions in G1-phase human cells

Repair of DNA double strand breaks (DSBs) is influenced by the chemical complexity of the lesion. Clustered lesions (complex DSBs) are generally considered more difficult to repair and responsible for early and late cellular effects after exposure to genotoxic agents. Resection is commonly used by the cells as part of the homologous recombination (HR) pathway in S- and G2-phase. In contrast, DNA resection in G1-phase may lead to an error-prone microhomology-mediated end joining. We induced DNA lesions with a wide range of complexity by irradiation of mammalian cells with X-rays or accelerated ions of different velocity and mass. We found replication protein A (RPA) foci indicating DSB resection both in S/G2- and G1-cells, and the fraction of resection-positive cells correlates with the severity of lesion complexity throughout the cell cycle. Besides RPA, Ataxia telangiectasia and Rad3-related (ATR) was recruited to complex DSBs both in S/G2- and G1-cells. Resection of complex DSBs is driven by meiotic recombination 11 homolog A (MRE11), CTBP-interacting protein (CtIP), and exonuclease 1 (EXO1) but seems not controlled by the Ku heterodimer or by phosphorylation of H2AX. Reduced resection capacity by CtIP depletion increased cell killing and the fraction of unrepaired DSBs after exposure to densely ionizing heavy ions, but not to X-rays. We conclude that in mammalian cells resection is essential for repair of complex DSBs in all phases of the cell-cycle and targeting this process sensitizes mammalian cells to cytotoxic agents inducing clustered breaks, such as in heavy-ion cancer therapy.     

Ileo-caecal resection induced pancreatic growth in rats

The effect of ileo-caecal resection on pancreatic growth was studied in rats four weeks after the operation. The results were compared with an identical control group who had undergone laparotomy alone. Pancreatic wet weight in ileo-caecal resectioned rats was 1.4 times greater than that found in control rats. Protein, DNA, RNA contents in the pancreas, pancreatic wet weight per 100 micrograms DNA and RNA/DNA ratio were also found significantly elevated in experimental group as opposed to the control group. Basal plasma levels of cholecystokinin (CCK) and gastrin were measured to delineate the influence of hormonal response on the pancreatic growth in ileo-caecal resected rats and were found not significantly increased after ileo-caecal resection. The data suggest that the enlargement of pancreas in ileo-caecal resected rats may be due to hyperplasia and hypertrophy of pancreatic cells; alternatively, the pancreatic growth may have been influenced by the bile acid deficiency and the reduction or release of an inhibitory factor present in the ileum of rats.     

内镜揭盖术与唇齿沟径路切除术治疗鼻前庭囊肿患者的疗效及对炎性因子的影响

目的:研究内镜揭盖术(ER)与唇齿沟径路切除术(LSPR)治疗鼻前庭囊肿(NVC)患者的临床疗效及对炎性因子的影响。方法:选择2015年7月至2017年6月来我院就诊的NVC患者106例,依据随机数字表法分别纳入ER组(53例)及LSPR组(53例)。ER组行ER治疗,LSPR组行唇齿沟径路切除术。观察两组术中出血量、手术时间、术后创口愈合时间、术后24 h疼痛评分及上皮化时间,两组术前、术后7 d白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)、C-反应蛋白(CRP)水平,两组术后并发症及复发情况。结果:ER组术中出血量、手术时间、术后创口愈合时间、术后24h疼痛评分及上皮化时间等手术指标均低于LSPR组,差异有统计学意义(P0.05);术后7 d,两组IL-6、IL-8、TNF-α及CRP水平均低于治疗前,且ER组低于LSPR组,差异有统计学意义(P0.05);ER组术后并发症发生率7.55%、术后复发率3.77%,均低于LSPR组的22.64%、20.75%,差异均有统计学意义(P0.05)。结论:与LSPR相比较,ER治疗NVC创伤小,恢复快,可有效降低术后炎性因子水平,术后并发症及复发率低,值得临床应用。     

超声引导下Mammotome微创旋切术治疗乳腺良性肿块

目的:研究超声引导下Mammotome微创旋切术对乳腺良性肿块的治疗价值。方法:回顾性分析2014年9月至2016年9月在本院就诊的BI-RADS分级为2~3级的387例乳腺良性肿块患者,运用Mammotome对729处乳腺病灶行微创旋切术,分析术后病理、并发症、随访半年后的治疗结果。结果:387例患者的729处乳腺病灶均获一次性成功切除。病理示均为良性病灶,其中32个合并不典型增生。术后出现局部血肿共11例(2.8%),皮下瘀斑共16例(4.1%),所有患者均未发生感染及气胸等严重并发症。术后6个月18个在病灶原部位发现肿块;手术无残留率为97.5%。结论:应用超声引导下Mammotome微创旋切术治疗乳腺良性肿块临床可行。     

七氟烷与丙泊酚对腹腔镜直肠癌根治术患者认知功能、T淋巴细胞及肝功能的影响

目的:探讨七氟烷与丙泊酚对腹腔镜直肠癌根治术患者认知功能、T淋巴细胞水平及肝功能指标的影响。方法:选择2015年6月-2017年8月期间于我院进行腹腔镜直肠癌根治术的110例患者为研究对象,按随机数字表法分为观察组和对照组,每组患者55例。观察组采用丙泊酚静脉全麻方式进行术前麻醉,对照组采用七氟烷吸入全麻方式进行术前麻醉。观察两组患者不同时间点平均动脉压(MAP)、心率(HR)情况,比较两组患者手术前后认知功能、T淋巴细胞水平、肝功能各项指标及不良反应发生情况。结果:两组患者不同时间点MAP、HR组间比较差异无统计学意义(P0.05)。术后6 h、12 h观察组简易精神状态量表(MMSE)评分低于术前,术后6 h、12 h、24 h对照组MMSE评分低于术前和观察组(P0.05)。术后3d两组患者CD3~+、CD4~+、CD4~+/CD8~+水平均降低,CD8~+水平均升高,且观察组CD3~+、CD4~+、CD4~+/CD8~+水平高于对照组,CD8~+水平低于对照组(P0.05)。术后3 d两组患者谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBILI)、直接胆红素(DBILI)水平均升高,但观察组患者各项指标水平低于对照组(P0.05)。观察组不良反应发生率为12.73%,与对照组的7.27%比较差异无统计学意义(P0.05)。结论:七氟烷与丙泊酚在腹腔镜直肠癌根治术中的麻醉效果相当,无严重不良反应发生,但应用丙泊酚进行麻醉对患者术后的认知功能、T淋巴细胞、肝功能指标的影响较小,值得临床推广应用。     

Immobilized Cyclodextrin Glycosyl Transferase for the Continuous Production of Cyclodextrins

Cyclodextrin glycosyl transferase (E.C: 2.4.1.19) from Bacillus, macerans and from a Bacillus sp. isolate was immobilized by two methods, viz. to epoxy-activated Sepharose and to alkylamine silica treated with glutaraldehyde. Because of the ready availability, low cost ($0.01/g), good surface area (30 M²/g) and ease of operation of a continuous cylindrical reactor, the high silica fabric was chosen. The immobilized enzyme had a pH optimum shifted to the alkaline side (from 6.5 to 7.5) and had a reduced temperature optimum (from 60°C to 50-55°C). Reuse efficiency showed 65% reduction in the overall activity of the immobilized enzyme after 10 cycles of 48 h each. Continuous operation at 55°C of a cylindrical reactor of 141 ml capacity, using the immobilized enzyme (80 g of high - silica fabric containing 114 mg of purified enzyme) gave a maximum productivity of 10.2 g of cyclodextrins L^-1 h,^-1, at a dilution rate of 0.32 h^-1 and a substrate concentration of 20 g L^-1. The half life of the biocatalyst was found to be 22 days, which could be further improved by using a lower operating temperature. Over the useful life time of the immobilized biocatalyst (22 days), the total Cyclodextrin produced was of the order of 88 Kg.     

内镜黏膜切除术治疗低位直肠侧向发育型肿瘤随机对照研究

目的:探讨内镜黏膜切除术治疗低位直肠侧向发育型肿瘤的临床效果。方法:收集我院收治的低位直肠侧向发育型肿瘤患者40例,随机分为对照组和实验组,每组各20例,对照组患者给予常规内镜下粘膜切除术,实验组患者给予内镜反转黏膜切除术,治疗结束后,对所有患者的病变残留例数、手术时间以及住院时间、并发症发生情况、手术切除效果进行检测并比较。结果:与对照组患者相比,实验组患者复发率较低(P0.05),手术时间以及住院时间、并发症发生率较低(P0.05);两组患者切除效果相比较,实验组分次切除、肿瘤残留例数患者较少,完全切除患者例数较多(P0.05),两组患者的整块切除例数无差异(P0.05)。结论:内镜反转黏膜切除术对于低位直肠侧向发育型肿瘤患者的临床疗效优于常规内镜下粘膜切除术,对临床有指导意义。     

The role of activated NLRP3 inflammatory body in acute kidney injury in rats caused by sepsis and NLRP3-TXNIP signaling pathway

ObjectiveThe model of acute renal injury (AKI) induced by sepsis in rats was established by abdominal resection through surgical suture. The activation mechanism of nod-like receptor with pyrin domain containing 3 (NLRP3) inflammatory corpuscle in AKI induced by sepsis was analyzed.MethodsHere, 60 male rats were selected and divided into two groups, including sham-operated group (NO-OPs group, n = 15) and sepsis group (CELP group, n = 45). In order to examine each index of CELP group, four time points (10, 20, 30, and 40 h) were set as control. In NO-OPs group, only abdominal resection through surgical suture was carried out. The expression levels of NLRP3, apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC), caspase-1, and the expression level of NLRP3-TXNIP signaling pathway were measured by immunohistochemistry, Western blotting, immunoprecipitation, and mito-TEMPO (a mitochondria-targeted antioxidant) 40 h after operation and 10, 20, 30, and 40 h post-operation in CELP group. Herein, 40 h post-operation in NO-OPs group and 10, 20, 30, and 40 h post-operation in CELP group, peripheral blood samples were collected.ResultsCompared with NO-OPs group, the levels of serum creatinine (Scr) and blood urea nitrogen (BUN) in CELP group were increased (P < 0.05). Compared with NO-OPs group, the expression levels of interleukin-1β (IL-1β), NLRP3, ASC, and caspase-1 in CELP group were increased (P < 0.05). The expression level of TXNIP in renal tubular epithelial cells in rats was up-regulated. There was a positive correlation between TXNIP and NLRP3. The binding of NLRP3-TXNIP signaling pathway could be inhibited by siRNA transfection or mito-TMPO, and the activity of NLRP3 inflammatory bodies could be inhibited as well.ConclusionActivation of NLRP3 inflammatory corpuscles could promote AKI induced by sepsis. Simultaneously, renal injury may lead to the production of mitochondrial reactive oxygen species (mROS), which may induce the binding of TXNIP to NLRP3.     

DNA End Resection: Nucleases Team Up with the Right Partners to Initiate Homologous Recombination

The repair of DNA double-strand breaks by homologous recombination commences by nucleolytic degradation of the 5′-terminated strand of the DNA break. This leads to the formation of 3′-tailed DNA, which serves as a substrate for the strand exchange protein Rad51. The nucleoprotein filament then invades homologous DNA to drive template-directed repair. In this review, I discuss mainly the mechanisms of DNA end resection in Saccharomyces cerevisiae, which includes short-range resection by Mre11-Rad50-Xrs2 and Sae2, as well as processive long-range resection by Sgs1-Dna2 or Exo1 pathways. Resection mechanisms are highly conserved between yeast and humans, and analogous machineries are found in prokaryotes as well.