玻璃化冻存延期回植技术在大面积皮肤撕脱伤中的应用研究

目的:探讨玻璃化冻存延期回植技术用于大面积撕脱皮肤伤的临床效果。方法:选择2015年6月至2018年6月收治的大面积皮肤撕脱伤的患者16例,根据处理方式分为撕脱皮肤玻璃化冻存二期回植组和撕脱皮肤一期回植组,每组8例患者。皮肤冻存后回植前,采用MTT法检测皮肤活力,比较两组皮片回植后的成活率、感染率、平均住院时间。结果:玻璃化冻存组皮肤冻存后平均活力为92.56%±7.20%,皮片成活率为81.74%±5.78%,感染率12.5%,平均住院时间33.13±3.91 d;一期皮肤回植组皮片成活率21.91%±5.28%,感染率100%,平均住院时间47.50±1.93 d。玻璃化皮肤保存组皮片成活率、感染率、平均住院时间均优于一期皮肤回植组(P0.05)。结论:皮肤玻璃化冻存延期回植技术用于无法一期回植皮肤撕脱伤的临床效果较好。     

Artificial selection,pre‐release diet,and gut symbiont inoculation effects on sterile male longevity for area‐wide fruit‐fly management

Longevity is an important life‐history trait for successful and cost‐effective application of the sterile insect technique. Furthermore, it has been shown that females of some species – e.g., Anastrepha ludens (Loew) (Diptera: Tephritidae) – preferentially copulate with ‘old’, sexually experienced males, rather than younger and inexperienced males. Long‐lived sterile males may therefore have greater opportunity to find and mate with wild females than short‐lived males, and be more effective in inducing sterility into wild populations. We explored the feasibility of increasing sterile male lifespan through selection of long‐lived strains and provision of pre‐release diets with added protein, and inoculated with bacterial symbionts recovered from cultures of the gut of wild Anastrepha obliqua (Macquart). Artificial selection for long‐lived A. ludens resulted in a sharp drop of fecundity levels for F1 females. Nevertheless, the cross of long‐lived males with laboratory females produced a female F1 progeny with fecundity levels comparable to those of females in the established colony. However, the male progeny of long‐lived males*laboratory females did not survive in higher proportions than laboratory males. Provision of sugar to A. obliqua adults resulted in increased survival in comparison to adults provided only with water, whereas the addition of protein to sugar‐only diets had no additional effect on longevity. Non‐irradiated males lived longer than irradiated males, and supplying a generic probiotic diet produced no noticeable effect in restoring irradiated male longevity of A. obliqua. We discuss the need to evaluate the time to reach sexual maturity and survival under stress for long‐lived strains, and the inclusion of low amounts of protein and specific beneficial bacteria in pre‐release diets to increase sterile male performance and longevity in the field.     

Effects of endogenous cannabinoid anandamide on cardiac Na+/Ca2+ exchanger

Endocannabinoid anandamide (N-arachidonoyl ethanolamide; AEA) has been shown to cause negative inotropic and antiarrhythmic effects in ventricular myocytes. In this study, using whole-cell patch clamp technique, we have investigated the effects of AEA on cardiac Na⁺/Ca²⁺ exchanger (NCX1)-mediated currents. AEA suppressed NCX1 with an IC₅₀ value of 4.7 μM. Both inward and outward components of exchanger currents were suppressed by AEA equally. AEA inhibition was mimicked by the metabolically stable analogue, methanandamide (metAEA, 10 μM) while it was not influenced by inhibition of fatty acid amide hydrolase with 1 μM URB597 incubation. The effect of AEA, was not altered in the presence of cannabinoid receptor 1 and 2 antagonists AM251 (1 μM) and AM630 (1 μM), respectively. In addition, inhibition by AEA remained unchanged after pertussis toxin (PTX, 2 μg/ml) treatment or following the inclusion of GDP-β-S (1 mM) in pipette solution. Currents mediated by NCX1 expressed in HEK-293 cells were also inhibited by 10 μM AEA a partially reversible manner. Confocal microscopy images indicated that the intensity of YFP-NCX1 expression on cell surface was not altered by AEA. Collectively, the results indicate that AEA directly inhibits the function of NCX1 in rat ventricular myocytes and in HEK-293 cells expressing NCX1.     

Technical guide for genetic advancement of underdeveloped and intractable Clostridium

In recent years, the genus Clostridium has risen to the forefront of both medical biotechnology and industrial biotechnology owing to its potential in applications as diverse as anticancer therapy and production of commodity chemicals and biofuels. The prevalence of hyper-virulent strains of C. difficile within medical institutions has also led to a global epidemic that demands a more thorough understanding of clostridial genetics, physiology, and pathogenicity. Unfortunately, Clostridium suffers from a lack of sophisticated genetic tools and techniques which has hindered the biotechnological exploitation of this important bacterial genus. This review provides a comprehensive summary of biotechnological progress made in clostridial genetic tool development, while also aiming to serve as a technical guide for the advancement of underdeveloped clostridial strains, including recalcitrant species, novel environmental samples, and non-type strains. Relevant strain engineering techniques, from genome sequencing and establishment of a gene transfer methodology through to deployment of advanced genome editing procedures, are discussed in detail to provide a blueprint for future clostridial strain construction endeavors. It is expected that a more thorough and rounded-out genetic toolkit available for use in the clostridia will bring about the construction of superior bioprocessing strains and a more complete understanding of clostridial genetics, physiology, and pathogenicity.     

‘灿烂’蓝莓组培与快繁技术

以蓝莓良种‘灿烂’半木质化茎段为材料,通过对增殖培养基以及生根培养基的筛选,建立蓝莓高效组培快繁技术体系。结果表明,最佳的增殖培养基为1/2MS＋2.0 mg·L^-1 ZT＋0.01 mg·L^-1 NAA＋30 g·L^-1白糖＋6.5 g·L^-1琼脂,芽诱导率为87%,增殖系数达3.6;ZT 1.0～2.0 mg·L^-1对不定芽诱导效果明显,随着ZT浓度的提高,愈伤组织产生越明显,影响增殖苗的质量。最佳生根培养基为1/2MS＋1.5 mg·L^-1 IBA＋0.01 mg·L^-1 NAA ＋25 g·L^-1白糖＋7.0 g·L^-1琼脂,pH值5.8,生根率70%,IBA浓度从0.5 mg·L^-1提高到1.5 mg·L^-1,生根越多,根系越壮,但也产生更多愈伤组织影响生根质量。炼苗10 d后移栽至草炭土,成活率78%。     

Carbon dioxide emissions through oxidative peat decomposition on a burnt tropical peatland

In Southeast Asia, a huge amount of peat has accumulated under swamp forests over millennia. Fires have been widely used for land clearing after timber extraction, thus land conversion and land management with logging and drainage are strongly associated with fire activity. During recent El Niño years, tropical peatlands have been severely fire‐affected and peatland fires enlarged. To investigate the impact of peat fires on the regional and global carbon balances, it is crucial to assess not only direct carbon emissions through peat combustion but also oxidative peat decomposition after fires. However, there is little information on the carbon dynamics of tropical peat damaged by fires. Therefore, we continuously measured soil CO₂ efflux [peat respiration (RP)] through oxidative peat decomposition using six automated chambers on a burnt peat area, from which about 0.7 m of the upper peat had been lost during two fires, in Central Kalimantan, Indonesia. The RP showed a clear seasonal variation with higher values in the dry season. The RP increased logarithmically as groundwater level (GWL) lowered. Temperature sensitivity or Q₁₀ of RP decreased as GWL lowered, mainly because the vertical distribution of RP would shift downward with the expansion of an unsaturated soil zone. Although soil temperature at the burnt open area was higher than that in a near peat swamp forest, model simulation suggests that the effect of temperature rise on RP is small. Annual gap‐filled RP was 382 ± 82 (the mean ± 1 SD of six chambers) and 362 ± 74 gC m^?2 yr^?1 during 2004–2005 and during 2005–2006 years, respectively. Simulated RP showed a significant negative relationship with GWL on an annual basis, which suggests that every GWL lowering by 0.1 m causes additional RP of 89 gC m^?2 yr^?1. The RP accounted for 21–24% of ecosystem respiration on an annual basis.     

A Xanthine-Derivative K+-Channel Opener Protects against Serotonin-Induced Cardiomyocyte Hypertrophy via the Modulation of Protein Kinases

This study investigated whether KMUP-1, a xanthine-derivative K⁺ channel opener, could prevent serotonin-induced hypertrophy in H9c2 cardiomyocytes via L-type Ca²⁺ channels (LTCCs). Rat heart-derived H9c2 cells were incubated with serotonin (10 μM) for 4 days. The cell size increased by 155.5%, and this was reversed by KMUP-1 (≥1 μM), and attenuated by the LTCC blocker verapamil (1 μM) and the 5-HT_2A antagonist ketanserin (0.1 μM), but unaffected by the 5-HT_2B antagonist SB206553. A perforated whole-cell patch-clamp technique was used to investigate Ca²⁺ currents through LTCCs in serotonin-induced H9c2 hypertrophy, in which cell capacitance and current density were increased. The LTCC current (I_Ca,L) increased ~2.9-fold in serotonin-elicited H9c2 hypertrophy, which was attenuated by verapamil and ketanserin, but not affected by SB206553 (0.1 μM). Serotonin-increased I_Ca,L was reduced by KMUP-1, PKA and PKC inhibitors (H-89, 1 μM and chelerythrine, 1 μM) while the current was enhanced by the PKC activator PMA, (1 μM) but not the PKA activator 8-Br-cAMP (100 μM), and was abolished by KMUP-1. In contrast, serotonin-increased I_Ca,L was blunted by the PKG activator 8-Br-cGMP (100 μM), but unaffected by the PKG inhibitor KT5823 (1 μM). Notably, KMUP-1 blocked serotonin-increased I_Ca,L but this was partially reversed by KT5823. In conclusion, serotonin-increased I_Ca,L could be due to activated 5-HT_2A receptor-mediated PKA and PKC cascades, and/or indirect interaction with PKG. KMUP-1 prevents serotonin-induced H9c2 cardiomyocyte hypertrophy, which can be attributed to its PKA and PKC inhibition, and/or PKG stimulation.     

General flexible nature of the cytosolic regions of fungal transient receptor potential (TRP) channels,revealed by expression screening using GFP‐fusion techniques

Transient receptor potential (TRP) channels are members of the voltage gated ion channel superfamily and display the unique characteristic of activation by diverse stimuli. We performed an expression analysis of fungal TRP channels, which possess relatively simple structures yet share the common functional characteristics with the other members, using a green fluorescent protein‐based screening methodology. The analysis revealed that all the tested fungal TRP channels were severely digested in their cytosolic regions during expression, implying the common flexibility of this region, as observed in the recent structural analyses of the fungal member, TRPGz. These characteristics are likely to be important for their diverse functions.