Systematic identification of chemical components in Fufang Shuanghua oral liquid and screening of potential active components against SARS-CoV-2 protease.

Coronavirus disease (COVID-19) caused by SARS-COV-2 infection has been widely prevalent in many countries and has become a common challenge facing mankind. Traditional Chinese medicine (TCM) has played a prominent role in this pandemic, and especially TCM with the function of "heat-clearing and detoxifying" has shown an excellent role in anti-virus. Fufang Shuanghua oral liquid (FFSH) has been used to treat the corresponding symptoms of influenza such as fever, nasal congestion, runny nose, sore throat, and upper respiratory tract infections in clinic, which are typical symptoms of COVID-19. The content of chlorogenic acid, andrographolide and dehydrated andrographolide as the quality control components of FFSH is not less than 1.0 mg/mL, 60 µg/mL and 60 µg/mL respectively. In this study, UPLC-Q-TOF-MS/MS was employed to describe the chemical profile of FFSH. Virtual screening and fluorescence resonance energy transfer (FRET) were used to screen the effective components of FFSH acting on SARS-CoV-2 main protease (Mpro). As a result, 214 compounds in FFSH were identified or preliminarily characterized by UPLC-Q-TOF-MS/MS, and 61 active ingredients with potential inhibitory effects on Mpro were selected through receptor-based and ligand-based virtual screening. In particular, quercetin, forsythoside A, and linoleic acid showed a good inhibitory effect on Mpro in FRET evaluation with IC50 values of 26.15 µM, 22.26 µM and 47.09 µM respectively, and had a strong binding affinity with the receptor Mpro (6LU7) in molecular docking. CYS145 and HIS41 were the main amino acid residues affected by small molecules in the protein binding domain. In brief, we characterized, for the first time, 214 chemical components in FFSH, and three of them, including quercetin, forsythoside A and linoleic acid, were screened out to exert beneficial anti-COVID-19 effects through CYS145 and HIS41 sites, which may provide a new research strategy for TCM to develop new therapeutic drugs against COVID-19.

Target-Based Virtual Screening and LC/MS-Guided Isolation Procedure for Identifying Phloroglucinol-Terpenoid Inhibitors of SARS-CoV-2.

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has led to more than 5 million deaths worldwide to date. Due to the limited therapeutic options so far available, target-based virtual screening with LC/MS support was applied to identify the novel and high-content compounds 1-4 with inhibitory effects on SARS-CoV-2 in Vero E6 cells from the plant Dryopteris wallichiana. These compounds were also evaluated against SARS-CoV-2 in Calu-3 cells and showed unambiguous inhibitory activity. The inhibition assay of targets showed that compounds 3 and 4 mainly inhibited SARS-CoV-2 3CLpro, with effective Kd values. Through docking and molecular dynamics modeling, the binding site is described, providing a comprehensive understanding of 3CLpro and interactions for 3, including hydrogen bonds, hydrophobic bonds, and the spatial occupation of the B ring. Compounds 3 and 4 represent new, potential lead compounds for the development of anti-SARS-CoV-2 drugs. This study has led to the development of a target-based virtual screening method for exploring the potency of natural products and for identifying natural bioactive compounds for possible COVID-19 treatment.

[Case-control study on the effect of meniscus shaping on knee function after anterior cruciate ligament reconstruction].

OBJECTIVE: To observe the effect of the meniscus shaping on the knee function and stability after anterior cruciate ligament reconstruction(ACLR). METHODS: A total of 64 ACLR patients were included from January 2013 to January 2015. The control group was the ACLR patients with intact meniscus, in which 24 males and 6 females. The mean age was(32.8±5.5) years old(ranged, 22 to 43 years old). The injury side was left on 17 cases and right on 13 cases. The mean follow-up time was(15.2±2.8) months(ranged, 12 to 19 months). The shaping group was the ACLR patients with meniscus shaping, in which 27 males and 7 females. The mean age was (33.1±4.2) years old (ranged, 23 to 42 years old). The injury side was on the left in 22 case and right in 12 cases. The mean follow-up time was (16.0±3.1) months (ranged, 12 to 20 months). The preoperative anterior tibia shift and knee joint function, as well as anterior tibia shift, knee joint function and active proprioception at last follow-up time were observed. The anterior tibia shift was measured by KT-1000. The knee joint function was assessed by Lysholm score and KOOS score. The errors of active proprioception were measured at 30°, 45° and 60° knee flexion. RESULTS: Postoperative anterior tibia shift of the affected side was (1.4±0.2) mm, which was lower than (2.2±0.4) mm in shaping group(P<0.05). The postoperative total Lysholm scores of the control group and the shaping group were 93.7±2.7 and 92.3±3.0 respectively, which were higher than 52.8±3.9 and 51.6±5.1 preoperatively(P<0.05), but there were no significant differences between two groups(P>0.05). In the KOOS score, the postoperative symptoms, pain, daily life, exercise capacity and life quality in control group were 90.7±5.5, 93.2±4.3, 96.8±2.2, 90.9±5.3, 91.8±4.5 respectively, which were higher than 72.7±6.0, 70.6±7.3, 72.5±7.4, 52.8±5.4, 36.2±6.5 preoperatively(P<0.05); the postoperative symptoms, pain, daily life, exercise capacity and life quality in the shaping group were 88.9±5.8, 92.6±3.5, 96.5±2.1, 89.3±7.2, 90.6±4.1 respectively, which were higher than 71.9±5.1, 71.2±7.1, 71.3±6.2, 53.1±6.1, 35.6±4.7 preoperatively(P<0.05). No significant differences were observed in each postoperative item of KOOS between the two groups(P>0.05). No significant differences were observed in the postoperative active proprioception error of contralateral side between the control group(12.2±3.4)°and shaping group(12.8±3.2)°(P>0.05). The error of active proprioception in the affacted side of the control group was(13.5±3.7)°, which was lower than that in the shaping group(17.1±4.2)°(P<0.05). In control group, there was no significant difference in the active proprioception error between two sides(P>0.05). While in shaping group, the error of active proprioception in the affacted side was significantly greater than that in the contralateral side(P<0.05). CONCLUSIONS: In short and medium term, meniscus shaping has no effect on knee joint function in patients with ACLR, but it impairs the knee proprioception and stability.

Improvement of FK506 production by synthetic biology approaches.

Synthetic biology has been applied to direct improvement of valuable metabolite productions. Tacrolimus (FK506), a clinically used immunosuppressive agent isolated from many Streptomyces, is produced by fermentation in industry. Here we chose FK506 as an example to review recent progress in improving FK506 production, including enhancing transcription levels of biosynthetic genes, accelerating post-translational modification levels of biosynthetic enzymes, increasing activities of rate limiting enzymes, and rational supplement of limited precursors. FK506 production was increased from 25 % to sevenfold by these synthetic biology approaches.

[Case control study on the treatment of acromioclavicular dislocation with Endobutton plates combined with an anchor].

OBJECTIVE: To compare the clinical effect of Endobutton plates combined with an anchor and clavicle hook plate in the treatment of acromioclavicular dislocation. METHODS: From January 2012 to August 2014, 83 patients with Rockwood type III acromioclavicular dislocation underwent surgical treatments. Among them, 34 patients were treated with Endobutton plate and anchor repair(Endobutton group), including 23 males and 11 females, and the mean age was(39.0±6.3) years old (26 to 51 years old); the average time from injury to operation was(4.1±1.3) days(3 to 7 days);the injured side:14 left, 20 right; the dislocation in 28 patients dues to fall, 6 patients dues traffic accident. There were 49 patients treated with clavicular hook plate(hook plate group), including 33 males and 16 females;the mean age was(37.9±6.3) years old (27 to 53 years old); the average time from injury to operation was(4.1±1.1) days (2 to 7 days);the injured side: 18 left, 31 right;the dislication in 36 patients dues to fall, 13 patients dues traffic accidents. The indexes such as intraoperative bleeding volume, operation time, incision size, postoperative complication and postoperative coracoclavicular space, shoulder joint function, and life quality were compared between two groups. RESULTS: In the hook plate group with 49 patients, the plates in 43 patients were removed at the secondary operation, and 32 patients had shoulder pain or limited active range. Thirty four patients in the Endobutton group had no pain symptoms and limited active range. All the patients did not suffer acromioclavicular dislocation again. There was no significant difference between the two groups in operation time, and intraoperative bleeding volume(P>0.05). The incision length in the hook plate group was longer than that in Endobutton group(P<0.05). The coracoclavicular space of the uninjured and injured side in two groups respectively had no significant differences, and the coracoclavicular space in the injured side between two group had no significant difference(P>0.05). There were no significant differences of Constant score and SF-36 between two groups 2 months after operation(P>0.05). Sixteen months after operation, the Constant score in the injured side of both groups was higher than that in 2 months postoperative. But the Constant score in the injured side of hook plate group was higher than that in Endobutton group(P<0.05). The Constant score in the uninjured side had no significant differences between two group(P>0.05). In hook plate group, the Constant score in the uninjured side was higher than that in the injured side. In Endobutton group, there were no significant differences of Constant score between two sides. The 16 month postoperative SF-36 in the injured side of both groups was higher than the 2 month postoperative one, but 16 month postoperative SF-36 in hook plate group was lower than that in Endobutton group (P<0.05). CONCLUSIONS: Endobutton plate combined with an anchor can effectively fix Rockwood type III or more acute acromioclavicular dislocation. The method has less complications, avoiding secondary removal of internal fixation.