In this study, we designed and synthesized 12 novel aloperine derivatives with different core structures. Among them, compound 3 with a ten-membered ring core was obtained through a special ring expansion reaction after γ-H Huffman elimination of quaternary ammonium salt, and the structure was verified by X-single crystal diffraction. Furthermore, their antiviral activity against human β-coronavirus HCoV-OC43 was evaluated by CCK-8 assay. Quaternary ammonium salt 2a and 3 had a good inhibitory effect against HCoV-OC43, and 2a had the highest anti-HCoV-OC43 activity with an EC₅₀ values of 3.77 μmol·L^-1 and a SI value of over 53.1. Schrӧdinger molecular docking results showed that both 2a and 3 might display their anti-HCoV-OC43 activity by directly acting on host TMPRSS2 and SR-B1. The results expanded the structural types of endocyclic aloperine and the function against coronavirus, and provided useful scientific data for the development of pharmaceutical applications of these compounds.

Objective: To understand the performance of 2019-nCoV nucleic acid detection in screening of contacts of COVID-19 cases in same flights and provide evidence for the effective screening of persons at high risk for the infection in domestic flights. Methods: The information of passengers who took same domestic flights with COVID-19 cases in China from April 1, 2020 to April 30, 2022 were retrospectively collected,and χ² test was used to analyze positive nucleic acid detection rates in the passengers in different times before the onsets of the index cases, in different seat rows and in epidemic periods of different 2019-nCoV variants. Results: During the study period, a total of 433 index cases were identified among 23 548 passengers in 370 flights. Subsequently, 72 positive cases of 2019-nCoV nucleic acid were detected in the passengers, in whom 57 were accompanying persons of the index cases. Further analysis of the another 15 passengers who tested positive for the nucleic acid showed that 86.67% of them had onsets or positive detections within 3 days after the diagnosis of the index cases, and the boarding times were all within 4 days before the onsets of the index cases. The positive detection rate in the passengers who seated in first three rows before and after the index cases was 0.15% (95%CI: 0.08%-0.27%), significantly higher than in the passengers in other rows (0.04%, 95%CI: 0.02%-0.10%, P=0.007),and there was no significant difference in the positive detection rate among the passengers in each of the 3 rows before and after the index cases (P=0.577). No significant differences were found in the positive detection rate in the passengers, except the accompanying persons, among the epidemics caused by different 2019-nCoV variants (P=0.565). During the Omicron epidemic period, all the positive detections in the passengers, except the accompanying persons, were within 3 days before the onset of the index cases. Conclusions: The screening test of 2019-nCoV nucleic acid can be conducted in the passengers took the same flights within 4 days before the onsets of the index cases on board. Passengers who seated within 3 rows from the index cases can considered as the close contacts at high risk for 2019-nCoV, for whom screening should be conducted first and special managements are needed. The passengers in other rows can be classified as general risk persons for screening and management.
Humans ; COVID-19 ; Retrospective Studies ; SARS-CoV-2 ; China ; Nucleic Acids

Oxidative Phosphorylation ; Acetylglucosamine ; Uridine Diphosphate N-Acetylglucosamine/metabolism*

Whitmania pigra is the most widely distributed species of leeches in the market. In this study, the effect of heavy metal lead pollution on the anticoagulant activity of Wh. pigra was studied and the potential mechanism was explored. Pb(NO_3)_2 was used to contaminate the breeding soil which was then used to rear Wh. pigra for 50 days(lead-contaminated group, LC group), and meanwhile the blank control group(CG group) was set. Proteins were extracted from the obtained leech samples, and the differentially expressed proteins between LC and CG groups were analyzed by label-free proteomics technology. In this study, a total of 152 differentially expressed proteins were screened out, of which 93 proteins were up-regulated and 59 proteins were down-regulated in LC group. Bioinformatics analysis showed that the biological processes enriched with the differentially expressed proteins were mainly vesicle-mediated transport and transport positive regulation; the enriched cell components were mainly endocytosis vesicles and apical plasma membrane; the enriched molecular functions mainly included carbohydrate binding. The differentially expressed proteins were enriched in 76 KEGG pathways, which mainly involved metabolic pathways, biosynthesis of secondary metabolites, and bacterial invasion of epithelial cells. In this study, two differentially expressed proteins with Antistasin domain were presumed, which provides reference for further exploring the regulatory mechanism and signal transduction underlying the effect of lead pollution on the anticoagulant activity of leech.
Animals ; Anticoagulants/pharmacology* ; Environmental Pollution ; Leeches ; Metals, Heavy ; Proteomics

Objective:To identify new host substrate of SseK3 and study its biological function.Methods:A yeast two hybrid system (Y2H) was used to identify the potential binding proteins of SseK3 from the Hela cDNA library; the arginine N-acetylglucosamine (Arg-GlcNAc) modification of the substrate protein by SseK3 was detected by co-expression in 293T cells and in vitro activity test; the modification sites of the substrate protein by SseK3 were detected by point mutation; the effect of Arg-GlcNAc modification of the substrate protein on its interaction protein binding ability was detected by immunoprecipitation test. Results:Results of Y2H and gene sequencing showed that Snapin was a new substrate of SseK3. Snapin could be Arg-GlcNAc-modified by SseK3 in vivo and in vitro; the modification sites of Snapin were arginine 119 and arginine 120; Arg-GlcNAc-modified Snapin inhibited its binding with SNAP25. Conclusions:Snapin, a new host substrate protein of SseK3, was successfully screened in this study. The Arg-GlcNAc modification of Snapin by SseK3 was studied, and the effect of this modification on Snapin function was preliminarily studied, which provided theoretical basis for further understanding the function of Arg-GlcNAc modification of bacteria and the mechanism of action in the process of pathogen infection.

Objective To investigate the spontaneous activity of brain neurons in patients with obsessive－compulsive disorder (OCD) under resting state. Methods Forty－eight OCD patients and 50 age－, gender－ and year of education－matched normal controls were enrolled. All the subjects underwent 3.0 T fMRI to acquire resting state brain image. The brain regions with significant differences in amplitude of low－frequency fluctuation (ALFF) between patients and controls were analyzed. Whole－brain functional connectivity (FC) were analyzed using the brain regions with significant differences as seed points, and the correlation between brain regions with significant differences in ALFF and FC analysis and obsessive－compulsive symptoms was analyzed. Results Compared to the control group, the ALFF of left dorsolateral prefrontal cortex increased in patients with OCD (t=4.305, P＜0.001). Compared to the controls, the analysis of whole－brain FC (based on MNI template) with the left dorsolateral prefrontal cortex as the regions of interest showed that the FC strength between the left dorsolateral prefrontal cortex and right orbital inferior frontal cortex (t=3.897, P＜0.001), left anterior cingulate cortex (t=3.370, P＜0.001), right anterior cingulate cortex (t=4.299, P＜0.001), left middle cingulate cortex (t=3.220, P＜0.001), right middle cingulate cortex (t=4.607, P＜0.001) enhanced; the FC strength between the left dorsolateral prefrontal cortex and the left opercular part of inferior frontal gyrus (t=－4.630, P＜0.001) weakened in patients with OCD. The FC between the left dorsolateral prefrontal cortex and the left opercular part of inferior frontal gyrus was negatively correlated with obsessions score(r=－0.369, P=0.014), compulsions score (r=－0.392, P=0.009) and total score (r=－0.393, P=0.008) of the Yale－Brown obsessive compulsive scale (Y－BOCS). Conclusion In patients with OCD, spontaneous neural activity of the left dorsolateral prefrontal cortex is enhanced in resting state, and the FC with multiple brain regions is abnormal. The FC strength between the left dorsolateral prefrontal cortex and the left opercular part of inferior frontal gyrus is associated with obsessive－compulsive symptoms.

Objective To investigate the clinical efficacy and safety of wedge osteotomy combined with internal fixation for ulnar olecranon fracture of Mayo type ⅡB.Methods From July 2014 to December 2017,8 patients with ulnar olecranon fracture of Mayo type ⅡB were treated with wedge osteotomy of the ulnar olecranon combined with internal fixation.They were 6 men and 2 women,from 28 to 45 years of age (mean,31.4 years).The wedge osteotomy was planned preoperatively according to the literature data and the osteotomy lengths were measured postoperatively to confirm the safe range for the surgery.Fracture healing was evaluated by regular X-ray follow-up and therapeutic effects were assessed by the range of elbow motion and the Broberg-Morry scoring system.Results On average in this group,the olecranon osteotomy was 6.2 mm in length on the articular sudace and 14.5 mm at the basal part.The olecranon was shortened by 4.6 cm on average after operation.All the 8 patients were followed up for 14 to 20 months (average,16.4 months).The clinical fracture healing time ranged from 12 to 16 weeks,with an average of 14.3 weeks.Follow-ups revealed no fracture displacement,pain or internal fixation failure.No significant differences were found between the affected and the contralateral sides in the flexion,extension or range of motion of the elbow joint at 3 months or at one year after surgery (P ＞ 0.05).According to the Broberg-Morry scoring system,5 cases were rated as excellent,2 as good and one as fair at 3 months after surgery.The final follow-ups found no traumatic arthritis or joint instability.Conclusions As a new treatment alternative,wedge osteotomy combined with internal fixation can obtain satisfactory curative outcomes for ulnar olecranon fractures of Mayo type ⅡB.The wedge osteotomy was performed in a safe range for this group.

In this study, the synthetic pathway of β-amyrin was constructed in the pre-constructed Saccharomyces cerevisiae chassis strain Y0 by introducing β-amyrin synthase from Glycyrrhiza uralensis, resulting strain Y1-C20-6, which successfully produced β-amyrin up to 5.97 mg·L~(-1). Then, the mevalonate pyrophosphate decarboxylase gene(ERG19), mevalonate kinase gene(ERG12), 3-hydroxy-3-methylglutaryl-CoA synthase gene(ERG13), phosphomevalonate kinase gene(ERG8) and IPP isomerase gene(IDI1)were overexpressed to promoted the metabolic fluxto the direction of β-amyrin synthesis for further improving β-amyrin production, resulting the strain Y2-C2-4 which produced β-amyrin of 10.3 mg·L~(-1)under the shake flask fermentation condition. This is 100% higher than that of strain Y1-C20-6, illustrating the positive effect of the metabolic engineering strategy applied in this study. The titer of β-amyrin was further improved up to 157.4 mg·L~(-1) in the fed-batch fermentation, which was almost 26 fold of that produced by strain Y1-C20-6. This study not only laid the foundation for the biosynthesis of β-amyrin but also provided a favorable chassis strain for elucidation of cytochrome oxidases and glycosyltransferases of β-amyrin-based triterpenoids.
Fermentation ; Glycyrrhiza uralensis ; enzymology ; genetics ; Industrial Microbiology ; Intramolecular Transferases ; genetics ; Metabolic Engineering ; Oleanolic Acid ; analogs & derivatives ; biosynthesis ; Saccharomyces cerevisiae ; metabolism

AIM To observe the effects of Qibao Meiran Oral Liquid (Polygoni multiflori Radix Praeparata,Angelicae sinensis Radix,Psoraleae Fructus,etc.) on learning and memory function,hippocampus tissue pathological morphology,SOD activity and carbonyl protein content in SAMP8 mice.METHODS Twenty-seven SAMP8 mice were randomly and equally divided into model control group,donepezil hydrochloride group and Qibao Meiran Oral Liquid group.Another nine SAMR1 mice were selected as normal control group.Mice were given successive intragastric administration for 60 days.On the 56th day,the passive avoidance test was adopted,and the learning and memory capacities were determined after 5 d;The pathological morphology was observed by HE staining;ELISA assay was used to detect the activity of SOD and the content of carbonyl protein in brain tissue.RESULTS Compared with the model control group,the escape latency of mice in the Qibao Meiran Oral Liquid group was significantly prolonged,and the number of errors decreased significantly (P ＜0.01);the pathological morphology of hippocampus tissue was significantly improved;SOD activity increased significantly,and carbonyl protein content decreased significantly (P ＜ 0.01).CONCLUSION Qibao Meiran Oral Liquid can not only improve the learning and memory function of SAMP8 mice,but also reduce the degree of hippocampus tissue degenerative disease.

Objective:To investigate the clinical features,diagnosis and surgical treatment of primary malignant tumors of the trachea and main bronchus.Methods:Clinical data of 35 cases with primary malignant tumors of the trachea and main bronchus in Zhongshan Hospital,Fudan University between January 2005 and December 2015 were analyzed retrospectively.Objective:The clinical symptoms were cough,sputum,bloody sputum,hemoptysis,shortness of breath,chest tightness,thoracalgia,and fever.There were 13 cases(37.1％)proved to be misdiagnosed in the early stage.All patients underwent surgical treatment without complications,and 3 patients were received postoperative radiotherapy.Among the 31 cases received postoperative follow-up,26 cases survived,5 cases died.The average disease-free interval was(34±16.2)months(16 to 49 months)and the median survival time was 50 months(16-138 months).Conclusions:Primary malignant tumors of the trachea and main bronchus without special clinical manifestations are easy to be misdiagnosed so that early bronchoscopy and chest CT examination are necessary.Active surgery treatment for primary malignant tumors of the trachea and main bronchus can achieve generate good prognosis.