Fluorescent and bimolecular-fluorescent protein tagging of genes at their native loci in Neurospora crassa using specialized double-joint PCR plasmids

The double-joint polymerase chain reaction (DJ-PCR) is a technique that can be used to construct vectors for targeted genome integration without laborious subcloning steps. Here we report the availability of plasmids that facilitate DJ-PCR-based construction of Neurospora crassa tagging vectors. These plasmids allow the creation of green or red fluorescent protein (GFP or RFP) tagging vectors for protein localization studies, as well as split-yellow fluorescent protein (YFP) tagging vectors for bimolecular fluorescence complementation (BiFC) analyses. We have demonstrated the utility of each plasmid with the tagging of known meiotic silencing proteins. Microscopic analysis of the tagged strains indicates that SMS-2 and QIP form macromolecular complexes in the perinuclear region during meiosis.     

Long COVID and its Management

The pandemic of COVID-19 is the biggest public health crisis in 21^st Century. Besides the acute symptoms after infection, patients and society are also being challenged by the long-term health complications associated with COVID-19, commonly known as long COVID. While health professionals work hard to find proper treatments, large amount of knowledge has been accumulated in recent years. In order to deal with long COVID efficiently, it is important for people to keep up with current progresses and take proactive actions on long COVID. For this purpose, this review will first introduce the general background of long COVID, and then discuss its risk factors, diagnostic indicators and management strategies. This review will serve as a useful resource for people to understand and prepare for long COVID that will be with us in the foreseeable future.     

Inhibition of miR146b-5p suppresses CT-guided renal cell carcinoma by targeting TRAF6

Renal cell carcinoma (RCC) is one of the most common malignancies in the urinary system. Due to the lack of early symptoms, diagnosis of RCC usually occurs at late stages or after cancer metastasis leading to poor prognosis. Therefore, it is crucial to study early molecular mechanisms and biomarkers. Previous studies have suggested that microRNAs are involved in RCC initiation and development, making them a good candidate for early diagnosis and therapy. MiR146b-5P plays important roles in the progression of multiple cancers including thyroid cancer, pancreatic cancer, cervical cancer. However, it is not clear whether and how miR146b-5P is involved in RCC. In this study, we aimed to investigate the function of miR146b-5P in RCC. We examined the expression levels of miR146b-5p in renal cancer tissue and cell lines. We also explored the effects of blocking miR146b-5p in renal tumor growth and inflammatory signaling. Finally, we determined if miR146b-5p regulates tumorigenesis through TRAF6. We found that miR146b-5p levels were significantly increased in renal cancer tissue and renal cancer cells. Blocking miR146b-5p suppressed renal tumor growth and enhanced inflammatory response through increased TRAF6 expression. These effects were eliminated in TRAF6 knockout mice. Our results suggest that enhanced miR146b-5p expression may be a biomarker for RCC and modulating miR146b-5p and TRAF6 levels represent a potential therapeutic strategy for RCC.     

NF-κB p65的敲减加重结肠上皮HCT116细胞氧化损伤

炎症细胞诱导的活性氧类生成和肠道氧化应激与慢性炎症性肠疾病以及结直肠肿瘤的发病密切相关。NF-κB信号通路参与氧化应激反应以及在结直肠炎症和肿瘤发生中的作用还并不完全清楚。本研究将化学合成一对编码小干扰RNA 序列、靶向人NF-κB 基因的长60 bp寡核苷酸链定向克隆至pSUPER小干扰RNA表达载体中,通过单酶切、双酶切及测序证实重组RNA干扰载体构建成功. 将构建成功的质粒转染至结肠上皮细胞HCT116中敲减p65,分别采用Western blot方法检测NF-κB p65蛋白表达水平,（3-(4,5-二甲基噻唑-2)-3,5-二苯基四氮唑溴盐（MTT）方法检测细胞存活情况. 结果显示,pSUPER-NF-κB p65载体可特异性下调NF-κB p65蛋白表达;下调p65表达可导致过氧化氢诱导的HCT116内活性氧类物质生成增高,存活细胞数目显著减少,氧化损伤加重。研究表明,在人结肠上皮细胞内NF-κB p65通路的抑制显著加重了结肠上皮细胞氧化损伤情况.     

Sodium (±)‐5‐bromo‐2‐(α‐hydroxypentyl) benzoate ameliorates pressure overload‐induced cardiac hypertrophy and dysfunction through inhibiting autophagy

Sodium (±)‐5‐bromo‐2‐(a‐hydroxypentyl) benzoate (generic name: brozopine, BZP) has been reported to protect against stroke‐induced brain injury and was approved for Phase II clinical trials for treatment of stroke‐related brain damage by the China Food and Drug Administration (CFDA). However, the role of BZP in cardiac diseases, especially in pressure overload‐induced cardiac hypertrophy and heart failure, remains to be investigated. In the present study, angiotensin II stimulation and transverse aortic constriction were employed to induce cardiomyocyte hypertrophy in vitro and in vivo, respectively, prior to the assessment of myocardial cell autophagy. We observed that BZP administration ameliorated cardiomyocyte hypertrophy and excessive autophagic activity. Further results indicated that AMP‐activated protein kinase (AMPK)‐mediated activation of the mammalian target of rapamycin (mTOR) pathway likely played a role in regulation of autophagy by BZP after Ang II stimulation. The activation of AMPK with metformin reversed the BZP‐induced suppression of autophagy. Finally, for the first time, we demonstrated that BZP could protect the heart from pressure overload‐induced hypertrophy and dysfunction, and this effect is associated with its inhibition of maladaptive cardiomyocyte autophagy through the AMPK‐mTOR signalling pathway. These findings indicated that BZP may serve as a promising compound for treatment of pressure overload‐induced cardiac remodelling and heart failure.     

洋葱伯克霍尔德菌致恶性肿瘤患者医院获得性感染的耐药性分析

目的分析我院恶性肿瘤患者医院获得性感染洋葱伯克霍尔德菌的临床特点及对常用抗菌药物的耐药性,为临床合理治疗提供依据。方法回顾性分析2011年1月至2013年12月,从我院恶性肿瘤感染患者送检的细菌培养标本;细菌鉴定用美国BD公司phoenix-100全自动细菌鉴定药敏系统,药敏试验采用纸片法,同时使用WHONET 5.6软件对相关资料进行统计。结果从检测部位分析主要分布在下呼吸道（74.1%）,其次为血液（9.4%）;药敏试验表明139株洋葱伯克霍尔德菌对米诺环素、氯霉素、美罗培南、头孢他啶和头孢哌酮/舒巴坦仍较敏感,可作为临床治疗洋葱伯克霍尔德菌感染的首选药物,其余15种抗菌药物的耐药率高达30.0%~100%。结论洋葱伯克霍尔德菌在恶性肿瘤患者中的耐药现象非常严重,临床应引起高度关注,及早进行微生物学检测,并根据药敏试验结果合理选用抗菌药物。     

陕南茶树栽培区域生态与地方品种分布

陕南秦巴山区是亚热带北缘地区。由于生态环境复杂,地区差异性大,茶树栽培与引种受到一定限制。大致以汉江谷地粮作区为界,南部大巴山北麓低山丘陵区为适栽区;北部秦岭南麓低山丘陵区则为次适栽区。地方茶树群体品种有7个,约由28个地方品种组成,呈斑、块状分布。