Association between Mitochondrial DNA Methylation and Hypertension Risk:A Cross-sectional Study in Chinese Northern Population

Hypertension is one of the most common chronic diseases worldwide and can considerably increase the incidence of complications such as stroke,coronary artery disease(CAD),heart failure,chronic kidney disease,aortic aneurysm,atrial fibrillation,and peripheral arterial disease.

RIG-I,a novel DAMPs sensor for myoglobin,activates NF-κB/caspase-3 signaling in CS-AKI model

Background:Acute kidney injury(AKI)is the main life-threatening complication of crush syndrome(CS),and myoglobin is accepted as the main pathogenic factor.The pattern recognition receptor retinoicacid-inducible gene I(RIG-I)has been reported to exert anti-viral effects function in the innate immune response.However,it is not clear whether RIG-I plays a role in CS-AKI.The present research was carried out to explore the role of RIG-I in CS-AKI.Methods:Sprague-Dawley rats were randomly divided into two groups:the sham and CS groups(n=12).After administration of anesthesia,the double hind limbs of rats in the CS group were put under a pressure of 3 kg for 16 h to mimic crush conditions.The rats in both groups were denied access to food and water.Rats were sacrificed at 12 h or 36 h after pressure was relieved.The successful establishment of the CS-AKI model was confirmed by serum biochemical analysis and renal histological examination.In addition,RNA sequencing was performed on rat kidney tissue to identify molecular pathways involved in CS-AKI.Furthermore,NRK-52 E cells were treated with 200μmol/L ferrous myoglobin to mimic CS-AKI at the cellular level.The cells and cell supernatant samples were collected at 6 h or 24 h.Small interfering RNAs(siRNA)was used to knock down RIG-I expression.The relative expression levels of molecules involved in the RIG-I pathway in rat kidney or cells samples were measured by quantitative real-time PCR(qPCR),Western blotting analysis,and immunohistochemistry(IHC)staining.Tumor necrosis factor-α(TNF-α)was d etected by ELISA.Co-immunoprecipitation(Co-IP)assays were used to detect the interaction between RIG-I and myoglobin.Results:RNA sequencing of CS-AKI rat kidney tissue revealed that the different expression of RIG-I signaling pathway.qPCR,Western blotting,and IHC assays showed that RIG-I,nuclear factor kappa-B(NF-κB)P65,p-P65,and the a poptotic marker caspase-3 and cleaved caspase-3 were up-regulated in the CS group(P<0.05).However,the levels of interferon regulatory factor 3(IRF3),p-IRF3 and the antiviral factor interferon-beta(IFN-β)showed no significant c hanges between the sham and CS groups.Co-IP assays showed the interaction between RIG-I and myoglobin in the kidneys of the CS group.Depletion of RIG-I could alleviate the myoglobin induced expression of apoptosis-associated molecules via the NF-κB/caspase-3 axis.C onclusions:RIG-I is a novel damage-associated molecular patterns(DAMPs)sensor for myoglobin and participates in the NF-κB/caspase-3 signaling pathway in CS-AKI.In the development of CS-AKI,specific intervention in the RIG-I p athway might be a potential therapeutic strategy for CS-AKI.

Clinical practice of emergency department-initiated extracorporeal cardiopulmonary resuscitation for cardiac arrest in adults

To the Editor:Cardiac arrest(CA)is one of the leading causes of adult death globally.According to the latest American Heart Association reports,[1]the incidences of Emergency Medical Service-assessed adult out-of-hospital cardiac arrest(OHCA)and in-hospital cardiac arrest(IHCA)were 140.7/100,000 and 10.2/1000,respectively.

Engineering of triterpene metabolism and overexpression of the lignin biosynthesis gene PAL promotes ginsenoside Rg3 accumulation in ginseng plant chassis

The ginsenoside Rgfound in Panax species has extensive pharmacological properties,in particular anti-cancer effects.However,its natural yield in Panax plants is limited.Here,we report a multimodular strategy to improve yields of Rgin a Panax ginseng chassis,combining engineering of triterpene metabolism and overexpression of a lignin biosynthesis gene,phenylalanine ammonia lyase(PAL).We first performed semi-rational design and site mutagenesis to improve the enzymatic efficiency of Pq3-O-UGT2,a glycosyltransferase that directly catalyzes the biosynthesis of Rgfrom Rh.Next,we used clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing to knock down the branch pathway of protopanaxatriol-type ginsenoside biosynthesis to enhance the metabolic flux of the protopanaxadiol-type ginsenoside Rg.Overexpression of PAL accelerated the formation of the xylem structure,significantly improving ginsenoside Rgaccumulation(to 6.19-fold higher than in thecontrol).Wecombinedoverexpression of the ginsenoside aglycon synthetic genes squalene epoxidase,Pq3-O-UGT2,and PAL with CRISPR/Cas9-based knockdown of CYP716A53v2 to improve ginsenoside Rgaccumulation.Finally,we produced ginsenoside Rgat a yield of 83.6 mg/L in a shake flask(7.0 mg/g dry weight,21.12-fold higher than with wild-type cultures).The highproduction system established in this study could be a potential platform to produce the ginsenoside Rgcommercially for pharmaceutical use.