Homo- or heterodimeric compounds that affect dimeric protein function through interaction between monomeric moieties and protein subunits can serve as valuable sources of potent and selective drug candidates. Here, we screened an in-house dimeric natural product collection, and panepocyclinol A (PecA) emerged as a selective and potent STAT3 inhibitor with profound anti-tumor efficacy. Through cross-linking C712/C718 residues in separate STAT3 monomers with two distinct Michael receptors, PecA inhibits STAT3 DNA binding affinity and transcription activity. Molecular dynamics simulation reveals the key conformation changes of STAT3 dimers upon the di-covalent binding with PecA that abolishes its DNA interactions. Furthermore, PecA exhibits high efficacy against anaplastic large T cell lymphoma in vitro and in vivo, especially those with constitutively activated STAT3 or STAT3^Y640F. In summary, our study describes a distinct and effective di-covalent modification for the dimeric compound PecA to disrupt STAT3 function.

Benzylisoquinoline alkaloids (BIAs) are a structurally diverse group of plant metabolites renowned for their pharmacological properties. However, sustainable sources for these compounds remain limited. Consequently, researchers are focusing on elucidating BIA biosynthetic pathways and genes to explore alternative sources using synthetic biology approaches. CYP80B, a family of cytochrome P450 (CYP450) enzymes, plays a crucial role in BIA biosynthesis. Previously reported CYP80Bs are known to catalyze the 3'-hydroxylation of (S)-N-methylcoclaurine, with the N-methyl group essential for catalytic activity. In this study, we successfully cloned a full-length CYP80B gene (StCYP80B) from Stephania tetrandra (S. tetrandra) and identified its function using a yeast heterologous expression system. Both in vivo yeast feeding and in vitro enzyme analysis demonstrated that StCYP80B could catalyze N-methylcoclaurine and coclaurine into their respective 3'-hydroxylated products. Notably, StCYP80B exhibited an expanded substrate selectivity compared to previously reported wild-type CYP80Bs, as it did not require an N-methyl group for hydroxylase activity. Furthermore, StCYP80B displayed a clear preference for the (S)-configuration. Co-expression of StCYP80B with the CYP450 reductases (CPRs, StCPR1, and StCPR2), also cloned from S. tetrandra, significantly enhanced the catalytic activity towards (S)-coclaurine. Site-directed mutagenesis of StCYP80B revealed that the residue H205 is crucial for coclaurine catalysis. Additionally, StCYP80B exhibited tissue-specific expression in plants. This study provides new genetic resources for the biosynthesis of BIAs and further elucidates their synthetic pathway in natural plant systems.
Cytochrome P-450 Enzyme System/chemistry* ; Benzylisoquinolines/chemistry* ; Hydroxylation ; Plant Proteins/chemistry* ; Alkaloids/metabolism* ; Stephania tetrandra/genetics*

COVID-19 is caused by coronavirus SARS-CoV-2. Current systemic vaccines generally provide limited protection against viral replication and shedding within the airway. Recombinant VSV (rVSV) is an effective vector which inducing potent and comprehensive immunities. Currently, there are two clinical trials investigating COVID-19 vaccines based on VSV vectors. These vaccines were developed with spike protein of WA1 which administrated intramuscularly. Although intranasal route is ideal for activating mucosal immunity with VSV vector, safety is of concern. Thus, a highly attenuated rVSV with three amino acids mutations in matrix protein (VSV_MT) was developed to construct safe mucosal vaccines against multiple SARS-CoV-2 variants of concern. It demonstrated that spike protein mutant lacking 21 amino acids in its cytoplasmic domain could rescue rVSV efficiently. VSV_MT indicated improved safeness compared with wild-type VSV as the vector encoding SARS-CoV-2 spike protein. With a single-dosed intranasal inoculation of rVSV_ΔGMT-S_Δ21, potent SARS-CoV-2 specific neutralization antibodies could be stimulated in animals, particularly in term of mucosal and cellular immunity. Strikingly, the chimeric VSV encoding S_Δ21 of Delta-variant can induce more potent immune responses compared with those encoding S_Δ21 of Omicron- or WA1-strain. VSV_MT is a promising platform to develop a mucosal vaccine for countering COVID-19.

Objective    To explore the relationship between myocardial viability in patients with coronary artery disease who underwent elective coronary artery bypass grafting (CABG) and early application of intra-aortic balloon pump (IABP) after coronary revascularization, and to provide relevant clinical reference for the pre-implantation of 16G single-lumen catheter in the femoral artery of high-risk patients to facilitate the addition of IABP after operation. Methods    This retrospective study included 521 patients (414 males and 107 females, aged 62.50±8.82 years) who underwent positron emission tomography (PET)-computed tomography (CT) perfusion-metabolism imaging prior to CABG surgery in our institution from December 2015 to August 2020. The myocardial viability information and left ventricular functional parameters were measured, including the proportion of non-viable myocardium (perfusion-metabolic imaging match), hibernating myocardium (perfusion-metabolic imaging mismatch) and dysfunctional myocardium (non-viable+viable myocardium), left ventricular ejection fraction, left ventricular end-diastolic volume and left ventricular end-systolic volume (LVESV). The patients were divided into an IABP group and a non-IABP group  according to whether they received IABP treatment after revascularization. The clinical data were reviewed and compared to explore significant impact factors between the two groups. And the multivariate logistic regression analysis was performed to investigate the correlation between preoperative myocardial viability and early use of IABP after CABG. Results    In multivariate logistic regression analysis, the amount of non-viable, dysfunctional myocardium and LVESV value were identified as the independent predictors for the probability of IABP use in the initial postoperative period. Receiver operating characteristic analysis showed that 9.5% non-viable myocardium, 19.5% dysfunctional myocardium, and LVESV of 114.5 mL were the optimal cutoff for predicting early IABP implantation during CABG. Conclusion    The myocardial survival status displayed by preoperative PET-CT myocardial perfusion-metabolism imaging can predict the possibility of applying IABP in CABG perioperative period. In addition to routine pre-anesthesia assessment, anesthesiologists can conduct risk stratification assessment for patients with CABG according to the results of preoperative myocardial viability imaging, which is of great significance to ensure the perioperative safety of high-risk patients with CABG.

Objective:To investigate the efficacy of treatment of Sanders Ⅱ & Ⅲ calcaneal fractures with an absorbable stick plus Kirschner wire through the tarsal sinus incision.Methods:From July 2017 to May 2020, 37 patients with 42 Sanders Ⅱ & Ⅲ calcaneal fractures were treated with an absorbable stick plus Kirschner wire through the tarsal sinus incision at The Third Ward of Department of Traumatic Orthopeadics, The Sixth Hospital of Ningbo. There were 25 males and 12 females, with an age of (48.2±5.6) years (from 20 to 69 years). The fractures were at the left side in 12 cases, at the right side in 20 and at bilateral sides in 5. By Sanders classification, 20 fractures were type Ⅱ and 22 ones type Ⅲ. Fracture union time and complications were recorded. Their B?hler and Gissane angles were compared between preoperation, postoperation and the last follow-up. The range of motion of the subtalar joint was evaluated by the Morrey method at 6 months postoperation. The functional recovery was evaluated by the American Society of Foot and Ankle Surgery (AOFAS) ankle-hindfoot score at 12 months postoperation.Results:The 37 patients were followed up for (15.2±2.7) months (from 13 to 18 months). There were no such complications as incision skin necrosis, Kirschner wire deformation, loss of fracture reduction or Kirschner wire infection. The anatomical morphology of the calcaneus was restored satisfactorily in the 37 patients. At preoperation, postoperation and the last follow-up, the B?hler angles were 13.3°±1.6°, 32.5°±5.5° and 32.7°±5.4° and the Gissane angles 78.3°±6.7°, 127.2°±6.7° and 128.0°±6.4°, respectively, showing significant differences between the preoperative and postoperative values ( P<0.05) but no significant differences between postoperation and the last follow-up ( P>0.05). The range of motion of the subtalar joint at 6 months postoperation was slightly limited in 25 cases and moderately limited in 12 cases, giving a rate of moderate and above limitation of 32.4% (12/37). By the AOFAS ankle-hindfoot score at 12 months postoperation, 12 cases were excellent, 21 ones good and 4 ones fair, giving a good to excellent rate of 89.2% (33/37). Conclusion:Treatment with an absorbable stick plus Kirschner wire through the tarsal sinus incision may lead to fine clinical efficacy for Sanders Ⅱ & Ⅲ calcaneal fractures.

Objective:To investigate the effect of motion correction (MC) on the calculated values of myocardial blood flow (MBF) and myocardial flow reserve (MFR) based on cadmium-zinc-telluride SPECT (CZT SPECT) images.Methods:Twenty-eight consecutive patients (10 males, 18 females, age: (60.75±11.62) years) with suspected or known coronary artery disease who underwent myocardial perfusion imaging (MPI) with dynamic CZT SPECT between June 2019 and August 2019 in TEDA International Cardiovascular Hospital were retrospectively analyzed. The MBF and MFR during rest imaging and stress imaging were quantitatively analyzed. Corridor 4DM software was used to calculate the stress MBF (sMBF) and MFR of the coronary artery branches and left ventricular (LV) before and after MC. The paired t test and Pearson correlation were used for data analysis. Results:The sMBF and MFR of LV before MC were (0.82±0.49) ml·min -1·g -1 and 1.69±0.68 respectively. After MC the two parameters increased to (1.05±0.64) ml·min -1·g -1 and 2.12±0.77 respectively ( t values: -4.87, -6.01, both P<0.001). The sMBF and MFR in left anterior descending (LAD), left circumflex (LCX), right coronary artery (RCA) and LV before MC were correlated with those after MC ( r values: 0.69-0.96, all P<0.001). If MFR <2.0 was used as the reference of impaired MFR, data before MC showed 19 patients (67.9%, 19/28) had impaired MFR, while 13 patients (46.4%, 13/28) had impaired MFR based on MFR values after MC. Conclusion:For MPI quantitative imaging with CZT SPECT, the calculated values of sMBF and MFR after MC is higher than those before MC, suggesting that MC is helpful to reduce the false positive results which may be caused by the " creep" effect of the heart.

ABSTRACT: Meningiomas are the most common primary intracranial neoplasm with diverse pathological types and complicated clinical manifestations. The fifth edition of the WHO Classification of Tumors of the Central Nervous System (WHO CNS5), published in 2021, introduces major changes that advance the role of molecular diagnostics in meningiomas. To follow the revision of WHO CNS5, this expert consensus statement was formed jointly by the Group of Neuro-Oncology, Society of Neurosurgery, Chinese Medical Association together with neuropathologists and evidence-based experts. The consensus provides reference points to integrate key biomarkers into stratification and clinical decision making for meningioma patients. REGISTRATION Practice guideline REgistration for transPAREncy (PREPARE), IPGRP-2022CN234.
Humans ; Meningioma/pathology* ; Consensus ; Neurosurgical Procedures ; Meningeal Neoplasms/pathology*

More than 85% of patients with uveal melanoma (UM) carry a GNAQ or GNA11 mutation at a hotspot codon (Q209) that encodes G protein α subunit q/11 polypeptides (Gα_q/11). GNAQ/11 relies on palmitoylation for membrane association and signal transduction. Despite the palmitoylation of GNAQ/11 was discovered long before, its implication in UM remains unclear. Here, results of palmitoylation-targeted mutagenesis and chemical interference approaches revealed that the loss of GNAQ/11 palmitoylation substantially affected tumor cell proliferation and survival in UM cells. Palmitoylation inhibition through the mutation of palmitoylation sites suppressed GNAQ/11^Q209L-induced malignant transformation in NIH3T3 cells. Importantly, the palmitoylation-deficient oncogenic GNAQ/11 failed to rescue the cell death initiated by the knock down of endogenous GNAQ/11 oncogenes in UM cells, which are much more dependent on Gα_q/11 signaling for cell survival and proliferation than other melanoma cells without GNAQ/11 mutations. Furthermore, the palmitoylation inhibitor, 2-bromopalmitate, also specifically disrupted Gα_q/11 downstream signaling by interfering with the MAPK pathway and BCL2 survival pathway in GNAQ/11-mutant UM cells and showed a notable synergistic effect when applied in combination with the BCL2 inhibitor, ABT-199, in vitro. The findings validate that GNAQ/11 palmitoylation plays a critical role in UM and may serve as a promising therapeutic target for GNAQ/11-driven UM.
Humans ; Mice ; Animals ; Lipoylation ; NIH 3T3 Cells ; Uveal Neoplasms/genetics* ; Melanoma/genetics* ; Cell Proliferation ; Proto-Oncogene Proteins c-bcl-2 ; GTP-Binding Protein alpha Subunits, Gq-G11/genetics*

Objective:To investigate the efficacy of the posterior axillary approach combined with the deltoid pectoralis major approach in the treatment of proximal humeral fracture combined with lower glenoid fracture.Methods:From July 2019 to September 2021, 7 patients were treated at Department of Traumatic Othopeadics, The Sixth Hospital of Ningbo for proximal humeral fracture combined with lower glenoid fracture by internal fixation via the posterior axillary approach combined with the deltoid pectoralis major approach. They were 2 males and 5 females, aged from 51 to 78 years (average, 62.9 years). All fractures were closed ones. According to the Neer classification for the proximal humeral fractures, there were one case of type Ⅱ, one case of type Ⅲ, 3 cases of type Ⅳ and 2 cases of type Ⅵ. According to the Ideberg classification for the glenoid fractures, 5 cases were type Ⅰ and 2 cases type Ⅱ. The anteroposterior, lateral and axillary X-ray films of the affected shoulder were taken at 6 and 12 weeks, and 6 and 12 months after operation to follow up fracture healing and occurrence of complications. The Constant-Murley shoulder joint scores and the Disability of Arm Shoulder and Hand (DASH) scores for the upper limb dysfunction were recorded at the last follow-up for all patients.Results:All the 7 patients were followed up for 8 to 15 months (mean, 11.9 months). Bone union was achieved after an average of 4.3 months (from 3 to 6 months) in all patients. None of the functional activities was affected in all by postoperative shoulder joint instability, incision infection or axillary scar hyperplasia. At the last follow-up, their Constant-Murley scores averaged 83.4 points (from 55 to 92 points), and their DASH scores 13.5 points (from 4.2 to 33.3 points).Conclusion:In the treatment of proximal humeral fracture combined with lower glenoid fracture, the posterior axillary approach combined with the deltoid pectoralis major approach can lead to fine early curative efficacy due to their advantageous possibilities to allow for easy fracture reduction, reliable fixation and early rehabilitation.

Objective:To investigate the diagnostic value of myocardial blood flow quantitative imaging with cadmium zinc telluride (CZT) SPECT in patients with high-risk coronary artery disease (CAD).Methods:A total of 148 patients (82 males, 66 females, age: (63.8±8.2) years) who successfully completed CZT SPECT dynamic acquisition and routine SPECT myocardial perfusion imaging (MPI) in TEDA International Cardiovascular Hospital from November 2018 to October 2020 were analyzed retrospectively. According to the results of coronary angiography (CAG), patients were divided into two groups: high-risk CAD group and low-to-medium-risk CAD group. At the case level, quantitative parameters (stress myocardial blood flow (sMBF), rest myocardial blood flow (rMBF) and myocardial flow reserve (MFR)), semi-quantitative parameters (summed stress score (SSS), summed rest score (SRS), summed difference score (SDS) and transient ischemic dilation (TID)) and left ventricular function parameters of two groups were compared. Diagnostic accuracy was evaluated by ROC curve analysis. At the vascular level, the correlation between the degree of coronary artery stenosis and some parameters was analyzed. Mann-Whitney U test, logistic regression, Spearman rank correlation analysis and DeLong test were used for statistical analysis. Results:Case level analysis showed that MFR and sMBF in high-risk CAD group were significantly lower than those in low-to-medium-risk CAD group (1.36(0.87, 1.64) vs 2.74(2.30, 3.33), 1.06(0.69, 1.48) vs 2.50(1.73, 2.95) ml·g -1·min -1; U values: 628.0 and 853.5, both P<0.001). MFR and SDS were independent predictors of high-risk CAD patients (odds ratio ( OR)=0.251(95% CI: 0.136-0.464), P<0.001; OR=1.188(95% CI: 1.026-1.375), P=0.021), and MFR was more capable of predicting high-risk CAD. MFR and sMBF had the highest accuracy in diagnosing high-risk CAD (AUCs: 0.885 and 0.844). Differences of AUCs between MFR and other parameters were statistically significant ( z values: 1.99-6.77, all P<0.05), and the best diagnostic cut-off value was ≤1.83 (sensitivity: 85.90%; specificity: 85.71%). Vascular level analysis showed that MFR and sMBF( R2 values: 0.39 and 0.35, both P<0.001) were negatively correlated with the degree of coronary stenosis, while SSS, SRS and SDS ( R2 values: 0.22, 0.12 and 0.14, all P<0.001) were positively correlated with the degree of coronary stenosis. Conclusion:Compared with conventional SPECT MPI, CZT SPECT myocardial blood flow quantitative imaging has better diagnostic efficacy and clinical value in patients with high-risk CAD.