Venetoclax (Ven) is now widely used for both acute myeloid leukaemia (AML) and high-risk myelodysplastic syndrome (MDS), yet there is no consensus on salvage regimens after Ven failure. This study retrospectively evaluated the efficacy and safety of selinexor combined with subcutaneous decitabine (DAC) in 10 patients with AML or MDS with excess blasts (MDS-EB1/2) who had experienced prior Ven treatment failure. A literature review was also performed. Among the 7 patients with AML, 1 achieved complete remission (CR), 2 achieved CR with incomplete hematologic recovery (CRi), 1 achieved partial remission (PR), 2 had no remission, and 1 experienced disease progression (PD). Among the 3 patients with MDS, 2 achieved marrow CR and 1 had stable disease (SD). The median duration of response among the 6 responding patients was 2 months (range, 0.5-6 months). All 10 patients experienced varying degrees of myelosuppression. Five patients had mild gastrointestinal reactions, all of which were manageable. The overall tolerability was good, and no treatment-related deaths occurred. These findings suggest that selinexor combined with subcutaneous decitabine offers a novel and well-tolerated therapeutic option for patients with myeloid malignancies who have previously failed venetoclax-based therapy.

Here we report the case of a CBF-AML patient with KIT p.D816V mutation who failed avapritinib induction therapy and subsequently underwent bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT), along with a literature review. The patient was a 64-year-old male who did not achieve remission after induction therapy with the DA regimen (Daunorubicin + Cytarabine). After reinduction with avapritinib combined with the DCHG regimen (Decitabine + Homoharringtonine + Cytarabine + Granulocyte colony-stimulating factor), he attained complete remission (CR) and flow cytometry minimal residual disease (MRD) negativity, but the RUNX1::RUNX1T1 fusion gene remained positive. During consolidation therapy, flow MRD reappeared, and the fusion gene level progressively increased. The patient then underwent a 9/10 HLA-matched unrelated donor allo-HSCT. Post-transplant, the fusion gene persisted, prompting treatment with the "ITI" regimen (with dose adjustments, including sequential addition of interferon and interleukin-2, pomalidomide incorporation, and standard vs. escalated dosing of "ITI" regimen). Currently, MRD negativity has been maintained for over 5 months, with good treatment tolerance. This finding suggest that the "ITI" regimen may serve as a novel and well-tolerated therapeutic option for CBF-AML patients with persistent RUNX1::RUNX1T1 fusion gene positivity after allo-HSCT and KIT p.D816V mutation, particularly in cases of avapritinib treatment failure.

Objective:To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of N-desalkylquetiapine, hydroxybupropion, N-desmethyl clozapine, N-desmethyl clomipramine, O-desmethyvenlafaxine and dehydro aripiprazole in human plasma.Methods:In June 2023, plasma samples were treated with methanol-acetonitrile (volume ratio of 1∶1) for protein precipitation and then detected by LC-MS/MS. It was separated by C18 column and eluted with 5 mmol/L ammonium acetate water and 5 mmol/L ammonium acetate methanol solution as mobile phase gradient. The metabolites of six psychotropic drugs were qualitatively and quantitatively by using electrospray positive ion multi-reactive ion monitoring scanning mode (MRM) .Results:The linear relationships of six psychotropic drug metabolites were good in the concentration range of 2-100 μg/L. The linear correlation coefficients were 0.9971-0.9999, the limits of quantification of the method were 0.10-6.00 μg/L, and the inter-and intra-day precision were 4.6%-9.8% and 1.5%-8.6%, with the recoveries of the spiked standards ranged from 90.0% to 106.1%.Conclusion:The LC-MS/MS method for the determination of metabolites of six psychotropic drugs in human plasma is simple, rapid and sensitive, and can be used for the qualitative and quantitative determination of metabolites in plasma samples of patients suspected of psychotropic drug poisoning.

Objective:To explore the efficacy and safety of B-Raf proto-oncogene, serine/threonine kinase (BRAF) inhibitor and epidermal growth factor receptor (EGFR) inhibitor combined with immune checkpoint inhibitor in microsatellite stable (MSS) BRAF V600E metastatic colorectal cancer (mCRC) patients.Methods:The data and outcomes of mCRC patients with MSS BRAF V600E who received BRAF inhibitor, EGFR inhibitor combined with immune checkpoint inhibitor in Tianjin Medical University Cancer Hospital from May 2022 to April 2024 were retrospectively collected.Results:A total of 12 mCRC patients were included in this study, the objective response rate was 50.0%, the disease control rate was 66.7%, and the median disease control time of patients who achieved objective response was 8.0 months. The median progression-free survival was 6.8 months and the median overall survival was 8.4 months. Overall adverse reactions were controllable, the most common treatment-related adverse events were fatigue (8 cases), fever (5 cases), and rash (4 cases). There were no grade 4 adverse event, serious adverse event, and treatment-related death.Conclusion:BRAF inhibitor and EGFR inhibitor combined with immune checkpoint inhibitor show good efficacy and controllable safety in BRAF V600E mCRC patients.

Objective:To evaluate the stability of percutaneous screw fixation for minimally invasive treatment of intra-articular calcaneal fractures using three-dimensional finite element analysis.Methods:CT scan was performed on the calcaneus of a normal adult for three-dimensional reconstruction. The DICOM data were imported into Mimics software to establish a model of a Sanders type IIB intra-articular calcaneal fracture. Based on the Essex-Lopresti classification of posterior facet morphology, the model was subdivided into two subtypes: tongue-type and depression-type. The calcaneus was divided into four fragments: sustentaculum tali, posterior tuberosity, anterior process (three points), and posterior articular surface (one surface). Two types of fixation methods, classical lateral anatomical plates and combinations of percutaneous screws, were simulated and performed. A three-dimensional finite element analysis was conducted by applying a stress combination of 420 N on the posterior subtalar articular surface, 200 N on the middle subtalar articular surface, and 300 N at the Achilles tendon insertion point. The maximum displacement and von Mises stress values of each bone fragment and implant were recorded to evaluate the biomechanical stability. For clinical validation, 34 patients with Sanders type IIB calcaneal fractures from Orthopedics Department of the Sixth Affiliated People's Hospital of Shanghai Jiao Tong University were treated with percutaneous reduction and screw fixation using the following configurations.Results:Under simulated stress, the A4 group with medial support screws in the tongue-type fracture subgroup demonstrated minimal overall calcaneal displacement (0.22 mm) and internal fixation displacement (0.14 mm). For the depression-type, the B2 group with medial support screws showed lower maximum stress in the calcaneus and internal fixation, at 22.04 MPa and 41.14 MPa, respectively, along with the lowest overall displacement (0.14 mm). The peak stress of all groups of implants remained below the material yield strength. The A4 and B2 protocols were applied to 15 cases of tongue-type calcaneal fractures and 19 cases of collapse-type calcaneal fractures. At the final follow-up The American Orthopaedic Foot & Ankle Society ankle-hindfoot score scale was 86.1±5.82 and 87.2±5.18, respectively, while the visual analog scale for pain was 1.60±1.24 and 1.58±1.02, respectively.Conclusions:Percutaneous screw fixation provided reliable stability for Sanders type IIB calcaneal fractures. The fixation configuration incorporating a medial support screw offers superior biomechanical performance in both tongue-type and depression-type fractures, representing an optimized minimally invasive technique with strong clinical applicability.

Objective:To evaluate the clinical value of multi-disciplinary treatment (MDT) combined with enhanced recovery after surgery (ERAS) for the elderly patients with osteoporotic ankle fracture.Methods:A retrospective analysis was conducted to analyze the 88 elderly patients with osteoporotic ankle fracture who had been treated with MDT combined with ERAS or non-MDT at Department of Foot and Ankle Surgery, Shanghai Sixth People's Hospital from January 2021 to January 2024. According to whether MDT was adopted or not, this cohort was assigned into 2 groups using the propensity score matching method: a MDT group and a non-MDT group with a matching ratio of 1∶1 (44 cases per group). The 2 groups were compared in terms of choice of intraoperative fixation, hospital stay, time for return to work/daily life, patient satisfaction questionnaire (PSQ-18) during hospitalization, ankle range of motion at 1 and 3 months after surgery, ankle-hindfoot score of American Orthopaedic Foot and Ankle Society (AOFAS), visual analogue scale (VAS) for pain, gait, and incidence of complications.Results:There were no significant differences in the preoperative general data between the 2 groups, indicating comparability ( P<0.05). The choice of intraoperative fixation, PSQ-18 [(78.4±8.5) points], AOFAS ankle-hindfoot score at 3 months after operation [(75.4±8.2) points], and gait in the MDT group were significantly better than those in the non-MDT group [(74.2±9.6) points and (70.9±9.4) points] ( P<0.05). There was no significant difference in the hospital stay or time for return to work/daily life between the 2 groups ( P>0.05). There was no statistically significant difference either in ankle dorsiflexion or plantarflexion, VAS for pain, or incidence of complications between the 2 groups at 1 or 3 months after surgery, as well as in AOFAS ankle-hindfoot score or gait at 1 month after surgery ( P>0.05). Conclusion:MDT combined with ERAS can effectively increase the therapeutic efficacy for the elderly patients with osteoporotic ankle fracture, improve their function of affected limbs, and enhance their patient satisfaction.

Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

Objective:To discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.

Objective:To investigate the effects of Yangrong Changchun Ointment on serum estradiol (E 2), follicle stimulating hormone (FSH), anti-Müllerian hormone (AMH), and on the Kiss peptin/protein one anti-G protein-coupled receptor 54 (GPR54) signaling pathway in rats with menopause syndrome (MPS). Methods:Totally 48 SD female rats were randomly divided into a sham-operation group ( n=8) and a modeling group ( n=40), and the rat model of menopausal syndrome was established by ovariectomy. After successful modeling, the rats were divided into the model group, Kunbao Pills group group, Yangrong Changchun Ointment high-, medium-, low-dosage groups according to random number table method, with eight rats in each group. Yangrong Changchun Ointment high-, medium-, low-dosage groups were orally administered with Yangrong Changchun Ointment suspension at doses of 10.0, 5.0, and 2.5 g/kg, while the Kunbao Pills group was orally administered with Kunbao Pills suspension at a dosage of 1 g/kg. The sham-operation group and model group were orally administered with distilled water of equal volume at a dosage of 1 ml/100 g body weight, once a day, for 28 consecutive days. The organ index of rats was calculated; the behavior of rats was determined by open filed test; HE staining was used to observe the histopathology of the uterus; ELISA was used to detect the levels of E 2, FSH, and AMH in the serum of rats; and qRT-PCR and Western blot were used to detect the mRNA and protein expressions of GPR54, Kiss-1, Phospholipase C (PLC), gonadotropin-releasing hormone (GnRH), and protein kinase C (PKC) in the hypothalamus. Results:Compared with the model group, the uterus, adrenal gland, spleen and thymus indices increased in the Yangrong Changchun Ointment high-dosage group ( P<0.05); serum levels of E 2 and AMH increased ( P<0.05), and the serum level of FSH decreased ( P<0.05); the mRNA and protein expressions of GPR54, Kiss-1, PLC, GnRH and PKC in the hypothalamus decreased ( P<0.05), and the uterine histopathological damage was improved. Conclusion:Yangrong Changchun Ointment can improve the pathological injury and estrogen level disorder in rats with MPS, and its mechanism may be related to inhibiting the activation of kisspeptin/GPR54 pathway.