Triterpenes, which have large application potential in the treatment of cancer, are the main active components of genuine medicinal material Alisma orientale (Sam.) Juzep. Farnesyl pyrophosphate synthase (FPPS) is one of the important rate-limiting enzymes in the synthetic pathway of triterpenes. In this study the FPPS full length cDNA of the A. orientale, was cloned via homology-based cloning approach and rapid amplification of cDNA ends (RACE). The full length of the FPPS cDNA was 1 531 bp (accession no. HQ724508), which contained a full 1 032 bp ORF that encoded 343 amino acids. The deduced protein sequence exhibited five conserved motifs, two of which is riched of Asp (DDXXD). The result of real-time quantitative PCR (QRT-PCR) showed that FPPS gene was expressed in different organs of A. orientale. The expression increased from October to the first ten-day period of December, and then decreased. The FPPS gene expression was higher in leaves but lower in leafstalk, tuber and root. HPLC analysis of active components 23-acetyl-alismol B of A. orientale. during different periods indicated that its change trend should be consistent with FPPS gene expression. It can be primarily deduced that FPPS gene should be an important control point in the synthetic pathway of Alisma terpenes. This study may facilitate the quality of medicinal plants through gene engineering in the future.

Objective To evaluate the clinical application of hs-cTnT in diagnosis of AMI. Methods The detectable rates of hs-cTnT and con-cTnT from 147 AMI ( including 122 NSTEMI )patients on immediate admission were compared. The related biological markers including hs-cTnT, con-cTnT, CKMB mass and MYO were determined for all samples from 481 patients with chest pain on immediate admission and 4 h, 12 h ,20 h and 28 h after admission. The receiver operating characteristic curve was used to evaluate the sensitivity and specificity of all markers. The change rates of hs-cTnT within 4 hours from AMI group, non-AMI heart disease group, AMI related high risk disease group and control group were compared with serial detection. Results The detection rates of hs-cTnT for AMI and NSTEMI patients were 90. 3% and 91.0%, and both were significantly higher than the rates of con-cTnT, which were 61.9% and 60. 6% (x2 =23.08,18. 64,all P＜0. 01 ). Among different makers obtained from different collecting times,hs-cTnT had the highest detection rate. For admission cases, the area under curve of hs-cTnT, con-cTnT,CKMB mass and MYO were 0.935, 0.851, 0.827 and 0.769 respectively, and the differences have statistical significance(Z1 = 3. 13, Z2 = 4. 46, Z3 = 5.62, all P ＜ 0. 05 ). Besides, there was a significant difference between the change rate of hs-cTnT of AMI and other groups (x2=166.09,P＜0. 01).Conclusions In comparison with con-cTnT, hs-cTnT could provide reliable results for earlier diagnosis of AMI, and could also reduce misdiagnosis and missed diagnosis of NSTEMI. Combining single test of hs-cTnT with serial tests was superior to using cut-off value alone in diagnosis. Moreover, it could be helpful to distinguish non-AMI patients from true AMI patients due to the improved detection sensitivity. Because of its good diagnostic performance, hs-cTnT test may limit the application value of some other "early markers".

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins