N-Linked Oligosaccharides of Aspergillus awamori Feruloyl Esterase Are Important for Thermostability and Catalysis

A unique N-linked glycosylation motif (Asn⁷⁹-Tyr-Thr) was found in the sequence of type-A feruloyl esterases from Aspergillus spp. To clarify the function of the flap, the role of N-linked oligosaccharides located in the flap region on the biochemical properties of feruloyl esterase (AwFAEA) from Aspergillus awamori expressed in Pichia pastoris was analyzed by removing the N-linked glycosylation recognition site by site-directed mutagenesis. N79 was replaced with A or Q. N-glycosylation-free N79A and N79Q mutant enzymes had lower activity than that of the glycosylated recombinant AwFAEA wild-type enzyme toward α-naphthylbutyrate (C4), α-naphthylcaprylate (C8), and phenolic acid methyl esters. Kinetic analysis of the mutant enzymes indicated that the lower catalytic efficiency was due to a combination of increased K _m and decreased k _cat for N79A, and to a considerably decreased k _cat for N79Q. N79A and N79Q mutant enzymes also exhibited considerably reduced thermostability relative to the wild-type.     

Association between the flap endonuclease 1 gene polymorphisms and cancer susceptibility: An updated meta-analysis

Flap endonuclease 1 (FEN1) has emerged as an important enzyme in the maintenance of genomic instability and preventing carcinogenesis. The relationship between FEN1 −69G>A (rs174538)+4150G>T (rs4246215) polymorphisms and cancer susceptibility has been reported; however, results were inconclusive. In the present study, a meta-analysis of data from eligible reports was carried out to summarize the possible relationship between FEN1 polymorphisms and cancer risk. A total of 11 articles, including 20 studies with 7366 cases and 9028 controls and 18 studies with 6649 cases and 8325 controls for FEN1 rs174538 and FEN1 rs4246215 polymorphisms, respectively, were recruited for meta-analysis. Overall, meta-analyses showed that FEN1 rs174538 and rs4246215 polymorphisms are significantly associated with the decreased risk of cancer. The stratified analysis proposed that both variants were associated with protection against gastrointestinal cancer, breast cancer, hepatocellular cancer, esophageal cancer, gastric cancer, colorectal cancer, and lung cancer. In conclusion, this meta-analysis revealed an association between FEN1 polymorphisms and cancer risk. Additional studies in a larger study population that include subjects from a variety of ethnicities are warranted to further verify our findings.     

股前外侧穿支皮瓣与胸腹带蒂皮瓣对手外伤组织缺损修复的应用效果及对创面愈合程度的影响

摘要 目的：探讨股前外侧穿支皮瓣与胸腹带蒂皮瓣对手外伤组织缺损修复的应用效果及对创面愈合程度的影响。方法：选取我院2018年12月到2020年12月共收治的119例手外伤组织缺损患者作为研究对象,随机分为2组,分别为对照组（n=59,应用胸腹带蒂皮瓣修复术）和观察组（n=60,应用股前外侧穿支皮瓣修复术）。对比两组患者治疗优良率,对比两组患者治疗前后手部创面面积、创面愈合程度以及组织愈合时间,对比两组患者治疗后的Jamar握力、TAM和DASH评分情况,对比两组患者的皮瓣成活率、皮瓣危象率和血管吻合时间。结果：通过对比两组患者治疗优良率发现,观察组患者优的人数为21例、良为35例,优良率为93.33%,对照组患者优的人数为16例,良为30例,优良率为77.97%,观察组高于对照组（P＜0.05）;治疗后,与对照组相比,观察组患者的手部创面面积、组织愈合时间和DASH评分显著减少,创面愈合程度以及TAM与Jamar握力显著增加（P＜0.05）;通过对比两组患者的皮瓣成活率、术后皮瓣危象率以及血管吻合时间发现,两组患者的术后皮瓣危象率、血管吻合时间对比无明显差异（P＞0.05）,两组患者的术后皮瓣成活率对比差异显著,观察组明显高于对照组（P＜0.05）。结论：对手外伤组织缺损患者应用股前外侧穿支皮瓣与胸腹带蒂皮瓣修复术均具有明显的修复效果,但是应用股前外侧穿支皮瓣能够提升治疗效果,提升患者创面愈合程度减少愈合时间,提升患者手部运动情况,提升术后皮瓣成活率,值得临床应用推广。     

Pedicle screw-based posterior dynamic stabilisation of the lumbar spine: in vitro cadaver investigation and a finite element study

Pedicle screw-based dynamic constructs either benefit from a dynamic (flexible) interconnecting rod or a dynamic (hinged) screw. Both types of systems have been reported in the literature. However, reports where the dynamic system is composed of two dynamic components, i.e. a dynamic (hinged) screw and a dynamic rod, are sparse. In this study, the biomechanical characteristics of a novel pedicle screw-based dynamic stabilisation system were investigated and compared with equivalent rigid and semi-rigid systems using in vitro testing and finite element modelling analysis. All stabilisation systems restored stability after decompression. A significant decrease in the range of motion was observed for the rigid system in all loadings. In the semi-rigid construct the range of motion was significantly less than the intact in extension, lateral bending and axial rotation loadings. There were no significant differences in motion between the intact spine and the spine treated with the dynamic system (P>0.05). The peak stress in screws was decreased when the stabilisation construct was equipped with dynamic rod and/or dynamic screws.     

Finite element analysis of Stryker Xia pedicle screw in artificial bone samples with and without supplemental cement augmentation

A validated, using in vitro biomechanical testing, finite element model was used to evaluate the affects of (1) cement augmentation and (2) an intact posterior cortex in osteoporotic bone. The presence of augmentation and/or a posterior cortical cortex increased the stabilization of the pedicle screw 2–5 fold. Placement of cement influenced failure load and toggle; with distal placement having the largest increase in failure load and decrease in cephalad–caudad toggle. The presence of posterior cortex caused a decrease in the amount of toggle, a proximal shift of the center of rotation and an increase in the maximum failure force.     

DNA repair mechanisms in dividing and non-dividing cells

DNA damage created by endogenous or exogenous genotoxic agents can exist in multiple forms, and if allowed to persist, can promote genome instability and directly lead to various human diseases, particularly cancer, neurological abnormalities, immunodeficiency and premature aging. To avoid such deleterious outcomes, cells have evolved an array of DNA repair pathways, which carry out what is typically a multiple-step process to resolve specific DNA lesions and maintain genome integrity. To fully appreciate the biological contributions of the different DNA repair systems, one must keep in mind the cellular context within which they operate. For example, the human body is composed of non-dividing and dividing cell types, including, in the brain, neurons and glial cells. We describe herein the molecular mechanisms of the different DNA repair pathways, and review their roles in non-dividing and dividing cells, with an eye toward how these pathways may regulate the development of neurological disease.     

后腹腔镜肾蒂淋巴管结扎术治疗乳糜尿(附46例报道)

目的:探讨后腹腔镜肾蒂淋巴管结扎术治疗乳糜尿的疗效,探讨微创治疗对乳糜尿的价值.方法:对2000年1月-2012年3月收治的92例乳糜尿患者随机分为实验组以及对照组,实验组患者采用后腹腔镜肾蒂淋巴管结扎术治进行治疗,对照组患者采用开放手术治疗,对两组患者手术时间、手术出血量、术后肠道恢复情况、拔管时间、术后下床活动时间、手术费用、住院肺炎以及住院天数、术后并发症的发生情况等进行比较.结果:实验组组手术时间为76.5±33.2 min、出血量为25.4±23.1mL、术后下床活动时间为39.8±5.6h、术后肠道恢复时间为33.5±5.4 h、手术住院天数为6.3± 2.4d、总住院天数为13.9±4.3 d、术后肺部感染发生例数为1例、术后切口感染发生例数为0例、术后体温大于38.5℃的例数为1例明显优于对照组的115.4± 23.2 min、187.3±56.7 mL、75.2±14.3 h、67.4± 15.4 h、12.3±4.5d、23.4± 7.3 d、5例、6例以及7例,实验组手术费用为7643±1453元高于对照组的3432±1043元,但实验组住院总费用为14854±2323元与对照组的14348±2134元无差异.结论:后腹腔镜肾蒂淋巴管结扎术对乳糜尿的治疗通过腹腔镜的放大使淋巴管结扎更完整同时其具有创伤小、术后住院时间短、术后无复发等特点.     

Cryopreservation of composite tissues and transplantation: preliminary studies

Despite advances in cryobiology, the reliable cryopreservation of complex tissues has not yet been achieved. This study evaluates the viability of cryopreserved composite flaps and demonstrates the feasibility of their transplantation. Epigastric flaps were harvested from male Lewis rats. 1.5 M dimethyl sulfoxide (Me₂SO) was used as the initial cryoprotectant agent (CPA). Samples were frozen at controlled rate to −140 °C and transferred to liquid nitrogen for at least two weeks. Hematoxylin and eosin (H/E) staining, MTT tetrazolium salt assay, and factor VIII immunostaining were used to evaluate the overall histology, epithelial viability, and vascular endothelial integrity, respectively, of cryopreserved flaps. For the in vivo phase, flaps were isotransplanted to 35 recipient animals, divided into three groups: fresh (n = 10), perfused (n = 8), and cryopreserved (n = 17). Blood vessel patency was assessed via Doppler at 1, 7, and 60 days post-transplantation. For in vitro studies, cryopreserved samples (10/10) retained normal cell architecture and vascular endothelial integrity upon H/E and factor VIII staining. The viability index of cryopreserved composite flap skin (n = 10) was 11.17 ± 2.01, which was not significantly different from fresh controls (n = 10, 12.15 ± 1.32). All transplanted flaps in the fresh and perfusion groups survived with healthy color and hair growth at 60 days after operation. Survival in the cryopreserved group ranged from 2 to 60 days, with a mean of 12 days. These results demonstrate that the long term survival of cryopreserved composite tissue transplants is possible. Further studies are needed to refine protocols for the reliable cryopreservation of composite parts.     

腓肠神经营养血管皮瓣修复跟骨骨折术后皮肤软组织缺损的临床研究

摘要 目的：探讨负压引流技术结合腓肠神经营养皮瓣在跟骨骨折钢板内固定术后皮肤软组织缺损的临床效果。方法：回顾性分析我院骨科2012年5月-2020年5月共31例跟骨骨折术后钢板外露,皮肤软组织缺损住院病人。纳入患者均使用负压引流技术结合腓肠神经营养皮瓣修复技术。创面给予彻底清创后行封闭负压吸引引流术,待创面新鲜后以腓肠神经营养皮瓣修复创面。对术后皮瓣成活情况;Maryland功能评分以及BMRC感觉功能评分进行综合评估。结果：术后2周时,28例皮瓣顺利成活,供区与受区伤口愈合良好,干燥、无渗出。3例术后出现皮瓣肿胀,皮瓣颜色发暗,伤口渗出较多,皮瓣边缘坏死,窦道形成等,给予切开引流、加强换药、敏感抗生素控制感染等治疗后,皮瓣成活。术后随访6-24个月皮瓣外观及功能恢复良好,无创面再坏死,裂开,感染等情况出现。其中2例再次入院行皮瓣整形术。术后6个月时,Maryland功能评分：优：17例;良：11例;优良率为：90.3%。BMRC感觉功能评分：S3-S4：20例;S2：8例;S1：3例。结论：腓肠神经营养皮瓣联合封闭负压吸引技术在跟骨骨折钢板内固定术后皮肤软组织缺损的治疗中能够缩短治疗时间,操作简单,疗效确切,可获得良好的修复效果。