柴油降解基因工程菌的构建及降解特性

【目的】构建柴油降解基因工程菌,提高柴油降解速率,研究p450基因在柴油降解过程中的作用。【方法】将Alcanivorax borkumensis SK2的p450基因合成后,连接至烷烃响应表达载体p Com8中,构建该基因的表达载体p450-SK2/p Com8,并将其转入大肠杆菌DH5α中,通过SDS-PAGE检测该基因在大肠杆菌DH5α中的表达,并将重组质粒p450-SK2/p Com8转入柴油降解菌Acinetobacter sp.Y9中,构建基因工程菌p450-SK2/Y9,研究工程菌p450-SK2/Y9对柴油的降解特性及p450基因在构建的工程菌p450-SK2/Y9中的表达。【结果】PCR、酶切及测序结果表明重组质粒p450-SK2/p Com8构建正确。当柴油诱导浓度大于1%时,目的基因在大肠杆菌DH5α中的蛋白表达量较大,且随着诱导时间的延长而呈增加趋势。通过PCR检测构建的基因工程菌p450-SK2/Y9中的p450基因表明,工程菌构建正确,利用单菌株降解柴油时,宿主菌Y9与工程菌p450-SK2/Y9的柴油降解效率未见明显差异,但工程菌p450-SK2/Y9在构建的菌群中对柴油降解的促进效果明显。SDS-PAGE结果表明,p450基因在构建的工程菌p450-SK2/Y9中能得到准确表达,在混合菌中的表达量高于单菌株。【结论】柴油降解基因工程菌在混合菌群中对柴油降解具有促进作用,而在单菌株情况下未见促进作用,且p450基因的蛋白表达在混合菌中也高于单菌株,这对于提高柴油的降解速率及研究p450基因在柴油降解过程中的作用机理具有一定意义。     

北京地区典型落叶阔叶乔木叶片含氮量和δ15N值对大气氮沉降的响应

为探究植物对大气氮沉降的响应和对这部分氮素的来源指示作用,本研究通过对北京地区198个采样点,典型落叶阔叶乔木杨属(Populus)和柳属(Salix)植物叶片进行采样,测定其叶片样品含氮量和δ~(15)N值。结果表明:北京地区杨属植物叶片含氮量为16.5—38.6g/kg,平均(24.0±4.0)g/kg;柳属植物叶片含氮量为17.2—36.2g/kg,平均(25.9±4.1)g/kg。研究区域范围内杨属、柳属植物叶片的含氮量均呈现出西北低、东南高的对角线型分布,与该区域大气氮沉降的空间变异相吻合。由于研究区域范围内气候因子无明显的变异,植物叶片的含氮量变化反应了大气氮沉降对植物元素化学计量特征的影响和植物对大气氮沉降的响应。北京地区杨属植物叶片δ~(15)N值为-3.95‰—8.10‰,平均(1.15±2.48)‰;柳属植物叶片δ~(15)N值为-3.04‰—9.73‰,平均(2.31±2.60)‰。杨属和柳属植物叶片的δ~(15)N值均呈现出西北高、中部高、东南低的空间分布,与叶片含氮量空间分布趋势相反。中部城区较高的δ~(15)N值反应了交通污染对大气含氮化合物增加的影响;西北部较高的δ~(15)N值反应了该区域受人为活动排放源的影响较少,自然的氮循环是其较高δ~(15)N值的主要原因;东南部较低的δ~(15)N值则有可能是由农业活动和交通共同作用的结果。     

泸沽湖流域土地利用方式对土壤肥力的影响

为探讨泸沽湖流域不同土地利用方式对土壤肥力的影响,采用野外调查、取样和室内分析相结合的方法,对泸沽湖流域内3种主要土地利用方式(农田、草地、林地)的土壤理化性状进行对比分析。结果表明:(1)土地利用类型对土壤肥力影响明显,农田含水率、全氮、速效氮、速效磷和速效钾含量相对较高;(2)林地有机质含量相对较高,草地全磷全钾含量相对较高;(3)土壤养分含量与土壤颗粒组成之间有一定的相关性,其中土壤全磷与砂粒呈显著正相关,土壤速效氮含量与粉粒呈显著负相关。     

Integration of a multicomponent intervention for hypertension into primary healthcare services in Singapore—A cluster randomized controlled trial

BackgroundDespite availability of clinical practice guidelines for hypertension management, blood pressure (BP) control remains sub-optimal (<30%) even in high-income countries. This study aims to assess the effectiveness of a potentially scalable multicomponent intervention integrated into primary care system compared to usual care on BP control.Methods and findingsA cluster-randomized controlled trial was conducted in 8 government clinics in Singapore. The trial enrolled 916 patients aged ≥40 years with uncontrolled hypertension (systolic BP (SBP) ≥140 mmHg or diastolic BP (DBP) ≥90 mmHg).Multicomponent intervention consisted of physician training in risk-based treatment of hypertension, subsidized losartan-HCTZ single-pill combination (SPC) medications, nurse training in motivational conversations (MCs), and telephone follow-ups. Usual care (controls) comprised of routine care in the clinics, no MC or telephone follow-ups, and no subsidy on SPCs. The primary outcome was mean SBP at 24 months’ post-baseline. Four clinics (447 patients) were randomized to intervention and 4 (469) to usual care. Patient enrolment commenced in January 2017, and follow-up was during December 2018 to September 2020. Analysis used intention-to-treat principles. The primary outcome was SBP at 24 months. BP at baseline, 12 and 24 months was modeled at the patient level in a likelihood-based, linear mixed model repeated measures analysis with treatment group, follow-up, treatment group × follow-up interaction as fixed effects, and random cluster (clinic) effects.A total of 766 (83.6%) patients completed 2-year follow-up. A total of 63 (14.1%) and 87 (18.6%) patients in intervention and in usual care, respectively, were lost to follow-up. At 24 months, the adjusted mean SBP was significantly lower in the intervention group compared to usual care (−3.3 mmHg; 95% CI: −6.34, −0.32; p = 0.03). The intervention led to higher BP control (odds ratio 1.51; 95% CI: 1.10, 2.09; p = 0.01), lower odds of high (>20%) 10-year cardiovascular risk score (OR 0.67; 95% CI: 0.47, 0.97; p = 0.03), and lower mean log albuminuria (−0.22; 95% CI: −0.41, −0.02; p = 0.03). Mean DBP, mortality rates, and serious adverse events including hospitalizations were not different between groups. The main limitation was no masking in the trial.ConclusionsA multicomponent intervention consisting of physicians trained in risk-based treatment, subsidized SPC medications, nurse-delivered motivational conversation, and telephone follow-ups improved BP control and lowered cardiovascular risk. Wide-scale implementation of a multicomponent intervention such as the one in our trial is likely to reduce hypertension-related morbidity and mortality globally.Trial registrationTrial Registration: Clinicaltrials.gov {"type":"clinical-trial","attrs":{"text":"NCT02972619","term_id":"NCT02972619"}}NCT02972619.

Tazeen H Jafar and colleagues present findings from a cluster-randomized controlled trial conducted to evaluate the effectiveness of an intervention designed to manage hypertension.     

Genome-wide CRISPR screen identifies synthetic lethality between DOCK1 inhibition and metformin in liver cancer

Metformin is currently a strong candidate anti-tumor agent in multiple cancers. However, its anti-tumor effectiveness varies among different cancers or subpopulations, potentially due to tumor heterogeneity. It thus remains unclear which hepatocellular carcinoma (HCC) patient subpopulation(s) can benefit from metformin treatment. Here, through a genome-wide CRISPR-Cas9-based knockout screen, we find that DOCK1 levels determine the anti-tumor effects of metformin and that DOCK1 is a synthetic lethal target of metformin in HCC. Mechanistically, metformin promotes DOCK1 phosphorylation, which activates RAC1 to facilitate cell survival, leading to metformin resistance. The DOCK1-selective inhibitor, TBOPP, potentiates anti-tumor activity by metformin in vitro in liver cancer cell lines and patient-derived HCC organoids, and in vivo in xenografted liver cancer cells and immunocompetent mouse liver cancer models. Notably, metformin improves overall survival of HCC patients with low DOCK1 levels but not among patients with high DOCK1 expression. This study shows that metformin effectiveness depends on DOCK1 levels and that combining metformin with DOCK1 inhibition may provide a promising personalized therapeutic strategy for metformin-resistant HCC patients.Supplementary InformationThe online version contains supplementary material available at 10.1007/s13238-022-00906-6.     

蜻蜓凤梨开花相关基因AfFPF1初步研究

FPF1 (flowering-promoting factor 1)是参与植物开花期遗传控制的重要家族之一。迄今为止,人们对蜻蜓凤梨中FPF1家族了解有限。本研究以热带花卉蜻蜓凤梨为材料,基于转录组测序数据,克隆获得AfFPF1基因。该基因编码的蛋白含103个氨基酸,分子质量为12.06 kD。AfFPF1转录本在蜻蜓凤梨的根中显著高表达,此外其表达量受外源乙烯强诱导,且响应赤霉素。AfFPF1基因在拟南芥中异位表达使其开花时间提前,莲座叶数目减少,促进其根系生长。对转基因拟南芥内源开花基因进行表达量检测,发现转基因植株中,一些开花促进基因如AtFT、AtAP1、AtLFY、AtFUL、AtSOC1表达量显著升高,而抑制开花基因AtFLC表达量下调,进一步证实AfFPF1能正调控拟南芥的开花时间,且可能与这些基因相互作用。研究结果初步证实AfFPF1可能参与调控蜻蜓凤梨开花过程,为进一步研究AfFPF1功能、通过基因工程调控蜻蜓凤梨开花以及乙烯诱导蜻蜓凤梨开花分子机制提供了理论依据。     

一株纤维蛋白溶解酶产生菌的鉴定及其产酶条件初步研究

从亚洲传统发酵食品--虾酱中筛选到一株产纤维蛋白溶解酶能力较强的菌株,通过形态和常规生理生化性质鉴定,发现该菌株与芽孢杆菌属细菌的特征很相近,结合16S rDNA序列分析,构建系统发育树,确定其分类地位,由中国典型培养物保藏中心定名为Bacillus sp.nov.SK006(CCTCC No.M 205071),并优化了发酵培养基组成及培养条件,本研究为该菌株的深入研究和广泛应用提供了理论依据.     

Alternatively activated macrophages at the recipient site improve fat graft retention by promoting angiogenesis and adipogenesis

The inflammatory response mediated by macrophages plays a role in tissue repair. Macrophages preferentially infiltrate the donor site and subsequently, infiltrate the recipient site after fat grafting. This study aimed to trace host‐derived macrophages and to evaluate the effects of macrophage infiltration at the recipient site during the early stage on long‐term fat graft retention. In our novel mouse model, all mice underwent simulated liposuction and were divided into 2 groups. The fat procurement plus grafting (Pro‐Grafting) group was engrafted with prepared fat (0.3 ml). The pro‐Grafting+M2 group was engrafted with prepared fat (0.3 ml) mixed with 1.0 × 10⁶ GFP+M0 macrophages, and then, 2 ng IL‐4 was injected into the grafts on Day 3. In addition, 1.0 × 10⁶ GFP+M0 macrophages were injected into the tail vein for tracing in the Pro‐Grafting group. As a result, GFP+macrophages first infiltrated the donor site and subsequently infiltrated the recipient site in the Pro‐Grafting group. The long‐term retention rate was higher in the Pro‐Grafting+M2 group (52% ± 6.5%) than in the Pro‐Grafting group (40% ± 3.5%). CD34+ and CD31+ areas were observed earlier, and expression of the adipogenic proteins PPAR‐γ, C/EBP and AP2 was higher in the Pro‐Grafting+M2 group than in the Pro‐Grafting group. The host macrophages preferentially infiltrate the donor site, and then, infiltrate the recipient site after fat grafting. At the early stage, an increase in macrophages at the recipient site may promote vascularization and regeneration, and thereby improve the fat graft retention rate.