A universal all-in-one RPA-Cas12a strategy with de novo autodesigner and its application in on-site ultrasensitive detection of DNA and RNA viruses.

Revolutionary all-in-one RPA-CRISPR assays are rapidly becoming the most sought-after tools for point-of-care testing (POCT) due to their high sensitivity and ease of use. Despite the availability of one-pot methods for specific targets, the development of more efficient methods for new targets remains a significant challenge. In this study, we present a rapid and universal approach to establishing an all-in-one RPA-Cas12a method CORDSv2 based on rational balancing amplification and Cas12a cleavage, which achieves ultrasensitive detection of several targets, including SARS-CoV-2, ASFV, HPV16, and HPV18. CORDSv2 demonstrates a limit of detection (LOD) of 0.6 cp/µL and 100% sensitivity for SARS-CoV-2, comparable to qPCR. Combining with our portable device(hippo-CORDS), it has a visual detection LOD of 6 cp/µL and a sensitivity up to 100% for SARS-CoV-2 and 97% for Ct<35 ASFV samples, surpassing most one-pot visual methods. To simplify and accelerate the process for new targets, we also develop a de novo autodesigner by which the optimal couples of primers and crRNA can be selected rapidly. As a universal all-in-one RPA-CRISPR method for on-site testing, CORDSv2 becomes an attractive choice for rapid and accurate diagnosis in resource-limited settings.

The Relationship of Methylenetetrahydrofolate Reductase Gene C677T Polymorphism and Ischemic Stroke in Chinese Han Population.

BACKGROUND: The purpose of this study was to investigate the relationship of methylenetetrahydrofolate reductase (MTHFR) gene C677T polymorphism and ischemic stroke Large-artery atherosclerosis (LAA) and Small-artery occlusion (SAO) subtypes in Chinese Han Population. MATERIALS AND METHODS: One hundred and thirty LAA patients and one hundred and fifty-nine SAO patients data were prospectively collected. The control group was one hundred and ninety-eight subjects from the Medical Examination Center in the same period. Plasma homocysteine (Hcy) levels were determined by a double reagent enzymatic cycling method. The genotypes of MTHFR polymorphism (C677T) were determined using polymerase chain reaction-restriction fragment length polymorphism. RESULTS: As compared with the controls, hypertension prevalence, smoking consumption ratio were significantly higher, plasma Hcy levels increased significantly, plasma high-density lipoprotein cholesterol levels were significantly reduced in LAA and SAO groups (P<0.05 for each). Elevated plasma Hcy levels were associated with MTHFR C677T polymorphism (P<0.01). The frequency of C/T+T/T genotypes and T allele in LAA and SAO groups was significantly higher than those in control groups (P<0.01). CONCLUSIONS: The present study suggests that hypertension, smoking, decreasing plasma high-density lipoprotein cholesterol levels and hyperhomocysteinemia (HHcy) are independent risk factors for LAA and SAO subtypes of ischemic stroke in Chinese Han Population. The MTHFR gene C677T C/T+T/T genotypes and T allele increases the risk of LAA and SAO subtypes of ischemic stroke.

Lipophilisation of Caffeic Acid through Esterification with Propanol Using Water-tolerable Acidic Ionic Liquid as Catalyst.

Propyl caffeate was synthesized to produce lipophilic antioxidant, which used caffeic acid and propanol as starting materials, acidic ionic liquid as catalyst. The highest yield of propyl caffeate (98.7±0.8%) have been achieved under the optimum as follows: 1-butylsulfonic-3-methylimidazolium tosylate showed the best catalytic performance, molar ratio of caffeic acid to propanol was 1:20, reaction temperature was 90°C and the amount of acidic ionic liquid was 40%. The relationship between temperature and the forward rate constant gave the activation energy of 33.6 kJ mol(-1), which indicated that acidic ionic liquid possesses high catalytic activity in the synthesis of PC. And the activity of acidic ionic liquid was not inhibited by the water produced during the esterification process. More importantly, this reaction system can even proceed smoothly when initial water content was 5%.

Chaenomeles speciosa: A review of chemistry and pharmacology.

Chaenomeles speciosa (Sweet) Nakai (C. speciosa, Rosaceae family) is an effective medicinal plant, which has long been used in China to treat various diseases, such as rheumatism, cholera, dysentery, enteritis, beriberi and vitamin C deficiency syndrome. A series of chemical constituents, including triterpenoid, phenolic and phenylpropionic acids, flavonoids, saccharides, essential oils and alkaloids, have been isolated from this plant and some have already been evaluated for their biological activities. Pharmacological investigations demonstrated that C. speciosa possesses anti-inflammatory, antinociceptive, antimicrobial, antioxidant, immunoregulatory, antiparkinsonian, hepatoprotective and antitumor properties. The objective of this review was to summarise available up-to-date and comprehensive information on C. speciosa and provide a relevant reference for further investigations.