The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines. Therefore, there remains a need for alternative antivirals that target processes less susceptible to mutations, such as the formation of six-helix bundle (6-HB) during the viral fusion step of host cell entry. In this study, a novel high-throughput screening (HTS) assay employing a yeast-two-hybrid (Y2H) system was established to identify inhibitors of HR1/HR2 interaction. The compound IMB-9C, which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity, was selected. IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion. It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex. In addition, virtual docking and site-mutagenesis results suggest that amino acid residues A930, I931, K933, T941, and L945 are critical for IMB-9C binding to HR1. Collectively, in this study, we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.

Research indicates that microbe activity within the human body significantly influences health by being closely linked to various diseases. Accurately predicting microbe-disease interactions (MDIs) offers critical insights for disease intervention and pharmaceutical research. Current advanced AI-based technologies automatically generate robust representations of microbes and diseases, enabling effective MDI predictions. However, these models continue to face significant challenges. A major issue is their reliance on complex feature extractors and classifiers, which substantially diminishes the models' generalizability. To address this, we introduce a novel graph autoencoder framework that utilizes decoupled representation learning and multi-scale information fusion strategies to efficiently infer potential MDIs. Initially, we randomly mask portions of the input microbe-disease graph based on Bernoulli distribution to boost self-supervised training and minimize noise-related performance degradation. Secondly, we employ decoupled representation learning technology, compelling the graph neural network (GNN) to independently learn the weights for each feature subspace, thus enhancing its expressive power. Finally, we implement multi-scale information fusion technology to amalgamate the multi-layer outputs of GNN, reducing information loss due to occlusion. Extensive experiments on public datasets demonstrate that our model significantly surpasses existing top MDI prediction models. This indicates that our model can accurately predict unknown MDIs and is likely to aid in disease discovery and precision pharmaceutical research. Code and data are accessible at: https://github.com/shmildsj/MDI-IFDRL.

Objective:To study the application of 3D printing technology in multi-center fenestrated/branched endovascular repair (F/B-EVAR) for endovascular repair of abdominal aortic diseases.Methods:From Feb 2018 to Mar 2023, The clinical and followup data of 316 cases of abdominal aortic lesions undergoing repair with F/B-EVAR at 69 medical centers nationwide using 3D printing technology to guide physician-modified stent graft were retrospectively analyzed.Results:The mean follow-up time of the patients was 23 months (2-60 months), and 24 cases were lost to follow up, the follow-up rate was 92.4% (292/316), the mean postoperative hospitalization time was (8.2±4.9) days. A total of 944 main abdominal branch arteries were reconstructed. Intraoperative reconstruction of 11 branches failed, with a success rate of 98.8% (933/944). Within 30 days after surgery, 8 patients died (2.5%), and 6 patients died during follow-up, a total of 14 patients died (4.4%). There were 11 cases (3.5%) of spinal cord ischemia and no patient suffered from permanent paraplegia. There were 19 patients (6.0%) with postoperative renal function injury. Internal leakage was found in 26 patients, and the rate of internal leakage was 8.2%.Conclusion:3D printing technology can accurately locate the location of branch arteries, simplifing the surgical process, shortening the learning curve , and improving clinical efficacy.

Objective:To evaluate the clinical application potential of a novel prostate specific membrane antigen (PSMA) targeted PET tracer 68Ga-PSMA-NYM032 in patients diagnosed initially with prostate cancer. Methods:A total of 63 patients (age (68.7±8.7) years) with suspected prostate cancers who received 68Ga-PSMA-NYM032 PET/CT imaging in Affiliated Hospital of Jiangnan University between March 2022 and January 2023 were enrolled prospectively. The diagnostic efficiency of 68Ga-PSMA-NYM032 PET/CT imaging was evaluated in a patient-centered manner. The ROI was drawn to obtain SUV max by semi-quantitative analysis with visual analysis, and the diagnostic threshold of SUV max was obtained by ROC curve analysis. The correlations of SUV max in primary foci with total prostate specific antigen (tPSA) and Gleason score (GS) were analyzed by Spearman rank correlation analysis. Based on the D′Amico risk stratification (prostate specific antigen (PSA)>20 μg/L and ≤20 μg/L, GS>7 and ≤7), the detection rates of metastases by 68Ga-PSMA-NYM032 PET/CT imaging in different stratifications were analyzed by Fisher exact test, and the differences between SUV max of metastases in different stratifications were determined by Mann-Whitney U test. Results:The accuracy of 68Ga-PSMA-NYM032 PET/CT imaging was 92.06%(58/63), the sensitivity was 96.55%(28/29), the specificity was 88.24%(30/34), the positive predictive value was 87.50%(28/32), the negative predictive value was 96.77%(30/31), and the optimal SUV max threshold was 6.9. 68Ga-PSMA-NYM032 showed varying degrees of high uptake in the primary foci of prostate cancer, and SUV max were positively correlated with tPSA and GS ( rs values: 0.657, 0.592, P values: <0.001, 0.001). Stratified analysis showed a statistically significant difference in the detection rate of bone metastases by 68Ga-PSMA-NYM032 PET/CT between the GS>7 and GS≤7 subgroups (9/17 vs 1/12; P=0.019), while no statistical significances were observed in the detection rates of bone metastases or lymph node metastases of another subgroups (all P>0.05). In addition, none of the differences in SUV max of metastases in patients with different stratifications were statistically significant ( z value: from -1.57 to -0.50, all P>0.05). Conclusions:68Ga-PSMA-NYM032 PET/CT imaging has good diagnostic efficacy for prostate cancer, and it may provide a new strategy for the precise diagnosis and treatment of prostate cancer. Besides, GS stratification may affect the detection rate of bone metastases by 68Ga-PSMA-NYM032 PET/CT imaging.

Klebsiella pneumoniae(K.pneumoniae)is an important pathogen that can cause severe hospital-and community-acquired infections.To systematically investigate its methylation features,we determined the whole-genome sequences of 14 K.pneumoniae strains covering varying serotypes,multilocus sequence types,clonal groups,viscosity/virulence,and drug resistance.Their methy-lomes were further characterized using Pacific Biosciences single-molecule real-time and bisulfite technologies.We identified 15 methylation motifs[13 N6-methyladenine(6mA)and two 5-methylcytosine(5mC)motifs],among which eight were novel.Their corresponding DNA methyl-transferases were also validated.Additionally,we analyzed the genomic distribution of GATC and CCWGG methylation motifs shared by all strains,and identified differential distribution pat-terns of some hemi-/un-methylated GATC motifs,which tend to be located within intergenic regions(IGRs).Specifically,we characterized the in vivo methylation kinetics at single-base resolu-tion on a genome-wide scale by simulating the dynamic processes of replication-mediated passive demethylation and MTase-catalyzed re-methylation.The slow methylation of the GATC motifs in the replication origin(oriC)regions and IGRs implicates the epigenetic regulation of replication initiation and transcription.Our findings illustrate the first comprehensive dynamic methylome map of K.pneumoniae at single-base resolution,and provide a useful reference to better understand epigenetic regulation in this and other bacterial species.

Objective:To investigate the clinical and genetic characteristics of cerebellar ataxia, neuropathy, vestibular areflexia syndrome (CANVAS) with replication factor C subunit 1 (RFC1) gene mutation to improve the understanding of this disease.Methods:A case of CANVAS diagnosed in the Peking University Third Hospital in January 2021 was reported. Detailed genetic analyses of ataxia were performed with DNA extracted from the peripheral blood of the patient. Studies including pathogenic variants of RFC1 gene causing CANVAS were reviewed and the clinical and genetic characteristics of the disease were summarized.Results:The patient was a 51-year-old female with the prominent manifestation of progressive walking instability. And the clinical data met the diagnostic criteria of CANVAS. The genetic tests excluded other hereditary ataxia mutations and identified the biallelic expansion of the pathogenic variant structure (AAGGG)exp repeat amplification in RFC1 gene. A total of 14 studies on CANVAS with RFC1 gene mutation were reviewed. The overall mutation rate of RFC1 gene in CANVAS was 68%-100%, and it varied in sporadic and familial CANVAS. And the mutation had ethnic differences.Conclusions:Among adult patients with late-onset ataxia, the combination of brain magnetic resonance imaging, electrophysiology tests and vestibular function examination is beneficial to the identification of CANVAS. And the genetic test of RFC1 gene has significant value in the diagnosis of this disease. This patient with CANVAS expands the disease spectrum of ataxia in China, and confirms that RFC1 gene mutation is of great significance in the screening of ataxia disorders in the Chinese population.

Objective:To explore the clinical effect of open reduction in the treatment of Tile C pelvic fracture combined with acetabular fracture in a specific sequence.Methods:Retrospectively analyzed the clinical data of 53 patients with Tile C type pelvic fracture combined with acetabular fracture from January 2014 to January 2019, and were divided into specific sequence group and non-specific sequence group according to the sequence of intraoperative reduction. A total of 29 cases were observed in the specific sequence group, including 20 males and 9 females; aged 43.8±14.8 years old (18-71 years), and the fractures were reduced in the sequence of "inside and out, then up and down" during the operation. There were 24 cases in the non-specific sequence group, including 14 males and 10 females; aged 44.4±14.7 years old (18-69 years), and fracture reduction was not performed in this sequence during the operation. According to the type of pelvic and acetabular fracture injury, we choose the appropriate position and surgical approach. After open reduction, the fracture was fixed with internal plants. The intraoperative blood loss, operation time, visual analogue scale (VAS) score were compared between the two groups. The quality of fracture reduction was evaluated by Matta score, pelvic fracture function recovery was evaluated by Majeed score, and acetabular fracture was evaluated by hip joint modified Merle d'Aubigné-Postel score.Results:There was no statistically significant difference in general data between the two groups before operation ( P>0.05), which was comparable. The intraoperative blood loss of the specific sequence group and the non-specific sequence group were 1 031.1±513.7 and 1 406.3±738.1 ml, and the operation time was 3.5±1.0 and 4.8±1.4 h; The differences between the two groups were statistically significant ( P<0.05). 53 patients were followed up for 14.8±1.6 months (12-18 months) after operation. The average postoperative VAS scores of specific sequence group and non-specific sequence group were 1.3±1.1 and 1.5±1.3 respectively, and there was no statistically significant difference. The effectiveness of the pelvic fracture Matta score standard was evaluated in the specific sequence group: excellent in 22 cases, good in 5 cases, fair in 2 cases, excellent and good rate was 93.1%; non-specific sequence group excellent in 10 cases, good in 6 cases, fair in 5 cases, poor in 3 cases, excellent and good rate was 66.7%, the difference was statistically significant ( P<0.05). Matta score of acetabular fracture: 21 cases were excellent in specific sequence group, 5 cases were good, 3 cases were poor, excellent and good rate was 89.7%; 9 cases were excellent in non-specific sequence group, 8 cases were good, 7 cases were poor, excellent and good rate was 70.8 %, the difference is statistically significant ( P<0.05). The results of the last follow-up pelvic fractures were evaluated by Majeed score: 20 cases were excellent in the specific sequence group, 7 were good, 2 were fair, excellent and good rate was 93.1%; 10 were excellent in the non-specific sequence group, 5 were good, 5 were fair, and 4 were poor, excellent and good rate was 62.5%, the difference was statistically significant ( P<0.05). At the last follow-up, the modified Merle d'Aubigné-Postel score was used to evaluate the efficacy: 20 cases were excellent in the specific sequence group, 5 were good, 4 were fair, the excellent and good rate was 86.2%; In the non-specific sequence group, 9 cases were excellent, 7 cases were good, 4 cases were fair, and 4 cases were poor, excellent and good rate was 66.7%, the difference was statistically significant ( P<0.05). During the follow-up period, none of the patients in the two groups developed fracture nonunion, heterotopic ossification, iatrogenic neurovascular injury, and femoral head necrosis. Trauma arthritis occurred in 4 patients in the non-specific sequence group. Conclusion:"Inside and out, then up and down" sequential reduction of Tile C pelvis combined with acetabular fracture can significantly shorten the operation time and reduce the amount of intraoperative blood loss. The surgical procedure is reasonable, which helps to improve the quality of fracture reduction and promote the functional recovery of patients.

Tocilizumab has been reported to attenuate the "cytokine storm" in COVID-19 patients. We attempted to verify the effectiveness and safety of tocilizumab therapy in COVID-19 and identify patients most likely to benefit from this treatment. We conducted a randomized, controlled, open-label multicenter trial among COVID-19 patients. The patients were randomly assigned in a 1:1 ratio to receive either tocilizumab in addition to standard care or standard care alone. The cure rate, changes of oxygen saturation and interference, and inflammation biomarkers were observed. Thirty-three patients were randomized to the tocilizumab group, and 32 patients to the control group. The cure rate in the tocilizumab group was higher than that in the control group, but the difference was not statistically significant (94.12% vs. 87.10%, rate difference 95% CI-7.19%-21.23%, P = 0.4133). The improvement in hypoxia for the tocilizumab group was higher from day 4 onward and statistically significant from day 12 (P = 0.0359). In moderate disease patients with bilateral pulmonary lesions, the hypoxia ameliorated earlier after tocilizumab treatment, and less patients (1/12, 8.33%) needed an increase of inhaled oxygen concentration compared with the controls (4/6, 66.67%; rate difference 95% CI-99.17% to-17.50%, P = 0.0217). No severe adverse events occurred. More mild temporary adverse events were recorded in tocilizumab recipients (20/34, 58.82%) than the controls (4/31, 12.90%). Tocilizumab can improve hypoxia without unacceptable side effect profile and significant influences on the time virus load becomes negative. For patients with bilateral pulmonary lesions and elevated IL-6 levels, tocilizumab could be recommended to improve outcome.
Antibodies, Monoclonal, Humanized ; COVID-19/drug therapy* ; Humans ; SARS-CoV-2 ; Treatment Outcome

Objective: To investigate the short-term clinical efficacy of laparoscopic common bile duct exploration(LCBDE) with primary suture or T tube drainage in the management of choledocholithiasis. Methods: The retrospective cohort study was conducted from January 2014 to December 2018 with the clinical data of patients with choledocholithiasis being analyzed. A total of 863 patients were enrolled in this study. There were 431 males and 432 females. The median age was 60 (range 11 to 94). These patients had received LCBDE with primary suture (n=287) and T tube drainage (n=576) in the Department of General Surgery, the First Affiliated Hospital of Nanjing Medical University. Observation indicators: (1)Preoperative blood biochemistry, including blood serum levels of total bilirubin, direct bilirubin, ALT, AST, GGT. (2) Intraoperative conditions, including operation time, blood loss, diameter of common bile duct, number of common bile duct stone.(3)Short-term postoperative conditions, including postoperative hospital stay, postoperative complications. Measurement data with non-normal distribution were described as M (P₂₅, P₇₅), and comparison between groups was done using Mann-Whitney U test. Comparison of count data between groups were analyzed using the chi-square test. Univariate analysis and subsequent multivariable logistic regression were used to investigated the factors affecting the selection of surgical methods. Results: (1) Blood serum levels of total bilirubin and direct bilirubin in the T tube drainage group were 17.0(12.2, 36.0) μmol/L, 7.6(4.9, 19.0) μmol/L, which were significantly higher than those of the primary suture group[15.7(11.8, 29.7) μmol/L, 6.7(4.4, 16.5) μmol/L)](Z=-2.023, -2.468, P<0.05). Preoperative blood serum levels of ALT, AST and GGT in the T tube drainage group were 56.7 (26.6, 128.8) U/L, 38.0(24.3, 75.8) U/L and 179.7(50.8, 394.4) U/L, the primary suture group were[68.2(24.8, 165.3) U/L, 35.5(22.8, 96.9) U/L and 235.2(74.9, 459.1) U/L], with no difference between the two groups (Z=-0.985, -0.437, -1.740, P>0.05). (2)The operation time of the primary suture group was 85(70, 100) min, which was significantly shorter than that of the T tube drainage group[97(75, 120) min](Z=-5.532, P<0.05). The diameter of common bile duct in the primary suture and T tube drainage group were 1.0(0.8, 1.2) cm and 1.0(0.8, 1.2) cm, respectively. Significant difference was observed between the two groups(Z=-2.071, P<0.05). The intraoperative blood loss in the primary suture and T tube drainage group were 20(10, 50) ml and 20(20, 50) ml, with no difference between the two groups (Z=-0.477, P>0.05). 61.32%(176/287) and 67.36%(388/576) of patients in the primary suture group and T tube drainage group were found with multiple stones in the common bile duct, with no difference between the two groups (χ²=3.083, P>0.05). (3)The primary suture group showed shorter postoperative hospital stay compared with the T tube drainage group[4(3, 5) d vs 6(5, 6) d, Z=-12.057, P<0.05]. The primary suture group showed more patients with bile leakage (2.09%) compared with that of the T tube drainage group (0.35%). Multivariable logistic regression showed that the number of common bile duct stone, diameter of common bile duct, time period of surgery, surgery group were significant factors affecting the selection of surgical methods(OR=1.687, 2.423, 0.587, 4.632, 95%CI: 1.152-2.470, 1.519-3.865, 0.511-0.675, 3.698-5.802, P<0.05). Conclusions Although different surgeons showed different opinions with the method of primary suture, laparoscopic common duct exploration with primary suture is safe and reliable in the management of choledocholithiasis with shorter operation time and faster postoperative recovery. T tube drainage is not absolutely necessary in the management of choledocholithiasis. Patients with multiple common bile duct stone or large diameter of common bile duct are likely to receive T tube drainage.

Objective Toexploretherelationshipbetweenthemicrostructuralintegrityofwhitematter(WM)andperipheralinterleukin10 (IL-10)inschizophrenia.Methods Diffusion MRIdataandvenousbloodsampleswereacquiredfrom47schizophreniapatients(SZ) and49healthycontrols (HC).Tract-basedspatialstatisticswasconductedtoexaminethedifferencesinFAandradialdiffusivity (RD)betweentwogroups.QuantitativechemiluminescenceassaywasperformedtomeasureperipheralIL-10levels.Regressionanalysiswas conductedtoinvestigatetherelationshipbetweenperipheralIL-10levelsanddiffusion measures (FAandRD).Results Compared withHC,therewerewidespreadreductionsinFAandincreaseinRDinSZ.Additionally,comparedwith HC,peripheralIL-10levels werehigherinSZ.PeripheralIL-10wasnegativelycorrelatedwithFAintherightposteriorthalamicradiationandleftinferiorfronto-occipitalfasciculusamongthepatients(β=-0.51,P=0.01andβ=-0.47,P=0.02,respectively)butnotcontrols(β=-0.01,P=0.95 andβ=-00.03,P=09.8,respectively).Andthecorrelationcoefficientsofthetwogroupsweredifferent(z=25.0,P=00.1andz=23.7,P=00.2, respectively).Conclusion TheperipheralIL-10maycontributetothedisruptionsofmicrostructuralWMintegrityinschizophrenia, supportingthenotionforanimportantroleofinflammationinthepathogenesisofschizophrenia.