OBJECTIVE: To establish venetoclax-resistant acute myeloid leukemia (AML) cell lines, assess the sensitivity of venetoclax-resistant cell lines to the BCL-2 protein family, and investigate their resistance mechanisms. METHODS: CCK-8 method was used to screen AML cell lines (MV4-11, MOLM13, OCI-AML2) that were relatively sensitive to venetoclax. Low concentrations of venetoclax continuously induced drug-resistance development in the cell lines. Changes in cell viability and apoptosis rate before and after resistance development were measured using the CCK-8 method and flow cytometry. BH3 profiling assay was performed to anayze the transform of mitochondrion-dependent apoptosis pathway as well as the sensitivity of resistant cell lines to BCL-2 family proteins and small molecule inhibitors. Real-time fluorescence quantitative PCR (RT-qPCR) was utilized to examine changes in the expression levels of BCL-2 protein family members in both venetoclax-resistant cell lines and multidrug-resistant patients. RESULTS: Venetoclax-resistant cell lines of MV4-11, MOLM13, and OCI-AML2 were successfully established, with IC₅₀ values exceeding 10-fold. Under the same concentration of venetoclax, the apoptosis rate of resistant cells decreased significantly (P < 0.05). BH3 profiling assay revealed that the drug-resistant cell lines showed increased sensitivity to many pro-apoptotic proteins (such as BIM,BID and NOXA). RT-qPCR showed significantly upregulated MCL1 and downregulated NOXA1 were detected in drug-resistant cell lines. Expression changes in MCL1 and NOXA1 in venetoclax-resistant patients were consistent with our established drug-resistant cell line results. CONCLUSION The venetoclax-resistant AML cell lines were successfully established through continuous induction with low concentrations of venetoclax. The venetoclax resistance resulted in alterations in the mitochondrial apoptosis pathway of the cells and an increased sensitivity of cells to pro-apoptotic proteins BIM, BID, and NOXA, which may be associated with the upregulation of MCL1 expression and downregulation of NOXA1 expression in the drug-resistant cells.
Humans ; Sulfonamides/pharmacology* ; Drug Resistance, Neoplasm ; Bridged Bicyclo Compounds, Heterocyclic/pharmacology* ; Leukemia, Myeloid, Acute/pathology* ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Cell Line, Tumor ; Apoptosis ; Antineoplastic Agents/pharmacology*

Objective To explore the therapeutic mechanism of Modified Lugen Formula(Phragmitis Rhizoma,Cicadae Periostracum,Batryticatus Bombyx,Lonicerae Japonicae Flos,Glycyrrhiza,Menthae Haplocalycis Herba,Notopterygii Rhizoma et Radix,Puerariae Lobatae Radix,Bupleuri Radix)in treating influenza from the virus-host interaction interface.Methods The phytocompounds were first collected from the HERB database,and then potential active compounds were screened out by Lipinski's rules of five.The targets of active compounds were further predicted through the SwissTargetPrediction platform.Differentially expressed genes(DEGs)were determined from the human H1N1 influenza dataset GSE90732 available in the Gene Expression Omnibus database(GEO).H1N1-Homo sapiens-related protein-protein interactions(PPIs)were gathered from the Pathogen-Host Interaction Search Tool(PHISTO).The above mentioned bioinformatic datasets were integrated.Then a PPI network and a Formula-virus-host interaction network were constructed using Cytoscape.Functional enrichment analyses were performed by using R software.Finally,molecular docking was carried out to evaluate the binding activities between the key compounds and targets.Results A total of 1 252 active compounds,1 415 targets,951 influenza-related DEGs,and 10 142 H1N1-Homo sapiens-related PPIs were obtained.There were 72 intersection targets between the Modified Lugen Formula and influenza.Functional enrichment analyses showed that these targets are closely related to host defense and programmed cell death.The network topological analysis showed that active compounds in the Modified Lugen Formula,such as oleanolic acid,γ-undecalactone,and longispinogenin,regulate viral proteins M2,NA,NS1,and HA and/or the host factors HSP90AA1,NRAS,and ITGB1,thus exert therapeutic effect.Molecular docking results confirmed that these compounds had a good binding ability with the targets.Conclusion Multiple active ingredients in Modified Lugen Formula directly target influenza virus proteins and/or host factors,thereby play an anti-influenza role in multiple dimensions,including inhibiting virus replication,regulating host defense and cell death.This study provides a theoretical basis for further experimental analysis of the action mechanism of the Modified Lugen Formula in treating influenza.

Objective:To evaluate the accuracy of the optical surface imaging system (OSI) using stereotactic radiosurgery (SRS) algorithm in single-center non-coplanar treatment of multiple brain metastases.Methods:Data of phantom and 15 patients with multiple brain metastases who underwent single-center non-coplanar radiotherapy in West China Hospital of Sichuan University from February to April 2022 were retrospectively analyzed. kV/MV and OSI imaging were used for imaging of the patients and phantoms under the same non-coplanar couch angle, respectively. The accuracy of OSI imaging of the phantoms and patients was evaluated using kV/MV imaging as reference image. The difference between the OSI and kV/MV systems is defined as accuracy, and the percentage of the absolute difference ≤1.00 mm in the translational direction or ≤0.50° in the rotational direction is defined as the threshold pass rate. Origin software was used to draw radar maps and Bland-Altman plots for statistical analysis.Results:When OSI images were used for the phantom imaging, the average differences in six-dimensional directions of lateral, long, vertical, rotational, roll and pitch were 0.03 mm, -0.09 mm, -0.27 mm, 0.04°, 0.17° and -0.19°, respectively. The maximum values were -2.20 mm, -2.30 mm, -1.20 mm, 0.60°, -1.00°, and -1.00°, respectively. When OSI system was utilized for the imaging of 15 patients, the average differences in six-dimensional directions were 0.44 mm, 0.16 mm, -0.20 mm, -0.11°, 0.10°, and -0.12°, respectively. The maximum values were -1.80 mm, 2.00 mm, 0.90 mm, -0.90°, -0.70°, and 0.80°, respectively. The translational errors mainly occurred in the lateral and long directions. The qualified rates of the threshold values of the phantoms and patients were 77% and 75% in the lateral direction, 82% and 89% in the long direction, respectively. In addition, 57% and 56% of patients met the threshold conditions of ±1.00 mm and ±0.50° in the six-dimensional directions, respectively.Conclusions:The OSI system using new SRS algorithm cannot meet the high accuracy requirements of single-center non-coplanar radiotherapy for multiple brain metastasis, especially in the lateral and long directions. It is not recommended for non-coplanar image guidance.

Objective:To compare the efficacy between Kirschner wire and clavicular hook plate in the fixation of distal clavicle fractures in older children.Methods:A retrospective analysis was conducted of the 28 pediatric patients with distal clavicle fracture who had undergone surgical treatment at Department of Pediatric Orthopedics, The Second General Hospital of Fuzhou from December 2014 to December 2023. There were 20 boys and 8 girls. Age: 10.0 (9.0, 12.8) years old; 22 left sides and 6 right sides; by the Craig modified Neer classification: 22 cases of type Ⅱ, 5 cases of type Ⅳ, and 1 case of type Ⅴ. The patients were divided into 2 groups based on their internal fixation methods. The Kirschner wire group of 12 cases were treated with open reduction and Kirschner wire fixation; the hook plate group of 16 cases were treated with open reduction and clavicular hook plate fixation. Operation time, hospital stay, fracture healing time, incidence of complications, and scores at the final follow-up (the Constant-Murley shoulder function score, quick disabilities of the arm, shoulder, and hand (quickDASH) score, and visual analog scale (VAS) for aesthetic satisfaction) were recorded and compared between the 2 groups.Results:There were no statistically significant differences in the baseline data between the 2 groups, indicating comparability ( P>0.05). The postoperative follow-up period for the 28 patients was (14.8±6.5) months. The operation time in the Kirschner wire group [(34.3±12.1) min] was significantly shorter than that in the hook plate group [(49.4±16.8) min] ( P<0.05), and the VAS score for aesthetic satisfaction at the final follow-up in the Kirschner wire group [(8.2±1.1) points] was significantly higher than that in the hook plate group [(6.2±2.6) points] ( P<0.05). There were no statistically significant differences between the 2 groups in terms of fracture healing time, hospital stay, incidence of complications, or Constant-Murley score and quickDASH score at the final follow-up ( P>0.05). Conclusions:For pediatric patients with distal clavicle fracture, both Kirschner wire fixation and clavicular hook plate fixation can achieve good functional outcomes. However, Kirschner wire fixation has advantages of shorter operation time, higher postoperative aesthetic satisfaction, and no need of a secondary surgery for implant removal.

Objective To explore clinical efficacy of F-type forceps for assisted reduction in femoral shaft fracture reduc-tion.Methods Forty-five patients with femoral shaft fracture treated with intramedullary nail and internal fixation from January 2019 to December 2021 were retrospectively analyzed and divided into two groups according to different reduction methods.In observation group,there were 21 patients,included 15 males and 6 females,aged from 27 to 92 years old with an average of(53.38±18.81)years old;9 patients on the left side,12 patients on the right side;7 patients were type A,8 patients were type B and 6 patients were type C according to AO fracture classification;the time from injury to operation ranged from 7 to 13 days with an average of(4.62±3.34)days;reduction was assisted by F-shaped forceps.In control group,there were 24 patients,in-cluding 17 males and 7 females,aged from 20 to 92 years old with an average of(51.96±20.43)years old;12 patients on the left side,12 patients on the right side;11 patients were type A,8 patients were type B and 5 patients were type C according to AO fracture classification;the time from injury to operation ranged from 2 to 13 days with an average of(6.29±3.04)days;tra-ditional reset mode was adopted.Operative time,intraoperative blood loss,intraoperative fluoroscopy times,intraoperative open reduction ratio,clinical healing time of fracture,postoperative complications,hospital stay,hospital cost and Lysholm score of knee joint at 6 and 12 months after surgery were compared between two groups to evaluate clinical effect.Results All patients were followed up for 12 to 24 months with an average of(16.60±3.45)months.In observation group,operative time,intraoper-ative blood loss,intraoperative fluoroscopy times,open reduction cases,and clinical healing time of fractures were(58.19±7.93)min,(88.10±44.45)ml,(25.29±5.54)times,0 case,(4.76±0.77)months,respectively;while in control group was(79.33±22.94)min,(222.92±144.45)ml,(47.46±26.25)times,5 cases,(7.13±1.80)months,and the difference between two groups were statistically significant(P＜0.05).There were no significant difference in postoperative complications,length of stay and hospitalization cost between two groups(P＞0.05).At 6 months after surgery,Lysholm score of knee joint in observa-tion group(88.62±4.48)was better than that in control group(79.21±8.91)(F=21.948,P=0.000).There were no significant difference in support use,pain and squat score between two groups(P＞0.05).At 12 months after surgery,Lysholm scores of stair climbing and pain in observation group were(9.62±1.20)and(19.76±1.92),which were better than those in control group(7.83±2.04)and(21.88±2.88)(P＜0.05).There were no significant difference in scores and total scores of other items between two groups(P＞0.05).Conclusion Compared with traditional reduction method,F-type forceps instrument could short-en operation time,reduce intraoperative blood loss,reduce intraoperative fluoroscopy times,accelerate clinical healing of frac-ture,and promote earlier functional recovery of knee joint.

Objective:To evaluate the accuracy and stability of stereotactic radiosurgery (SRS) algorithm in optical surface imaging (OSI) system in non-coplanar radiotherapy.Methods:Three OSI imaging systems were used to measure the phantom repeatedly at different couch rotation angles to analyze the accuracy and stability of OSI system. Seven patients with multiple brain metastases who underwent single-center non-coplanar radiotherapy were randomly selected, and the accuracy and stability of OSI for patient imaging were analyzed. Stability is defined as the difference between the two OSI measurements when the couch is turned from 0° to a non 0° angle, and then back to 0°, using the 0° cone beam CT (CBCT) as the "gold standard". Accuracy is defined as the difference between OSI and CBCT (at 0° couch angle) measurement data. The measurement data with normal distribution were described as Mean ± SD. The data with non-normal distribution were expressed as M (Q). The difference of the former data was compared by one-way ANOVA, and the difference of the latter data was assessed by Kruskal-Wallis H nonparametric test. Results:For non-coplanarity, the translation accuracy of the phantom and the patient was ≤ 1.30 mm and ≤ 1.00 mm, and the rotation accuracy was ≤ 0.50° and ≤ 0.60°, respectively. The translation errors mainly occurred in the left-right and head-foot directions. In terms of stability, the maximum standard deviation of phantom coplanar translation and rotation was 0.06 mm and 0.06°. The maximum standard deviation of patient translation and rotation was 0.17 mm and 0.19°.Conclusions:Although the new SRS algorithm improves the non-coplanar accuracy, it still cannot meet the precise requirements of non-coplanar single isocenter radiotherapy for multiple brain metastases, especially in the left-right and head-foot directions. When the couch rotation angle is large, OSI is not recommended for image-guided radiotherapy. However, its high stability can be used to monitor the intrafractional motion of patients.

Dihydrofolate reductase (DHFR) is a well-known key target in the treatment of tumors, bacterial infections, and parasitic infections; and it plays a critical role in the biosynthesis of cellular DNA. DHFR inhibitors interfere with one-carbon metabolism by inhibiting substrate binding to DHFR, thereby inhibiting cell proliferation. Research on DHFR inhibitors has continued since the 1940s. To date, a variety of DHFR inhibitors have come into the market, primarily used for anti-tumor, antibacterial, antiparasitic, and anti-inflammatory therapy. This review summarizes the research progress of DHFR inhibitors with antitumor or antibacterial effects in recent years based on the classification of single-target and dual-target and looks forward to the opportunities and challenges faced by the work in this field.

[This corrects the article DOI: 10.1016/j.apsb.2022.02.031.].

Objective: To investigate the morphology and immunohistochemical (IHC) expression of pseudostratified ependymal tubules in ovarian mature teratoma (MT). Methods: Five cases of ovarian MT with pseudostratified ependymal tubules were collected from Shenzhen Hospital(Futian) of Guangzhou University of Chinese Medicine and the Eighth Affiliated Hospital of Sun Yat-sen University from March 2019 to March 2022. In addition, 15 cases of ovarian MT with monolayer ependymal epithelium from Shenzhen Hospital (Futian) of Guangzhou University of Chinese medicine and seven cases of immature teratoma (IMT) from Hainan Provincial People's Hospital from March 2019 to March 2022 were collected as control. The morphologic characteristics and immunophenotypes of pseudostratified ependymal tubules, monolayer ependymal epithelium, and primitive neural epithelial tubules were observed and compared by H&E stain and IHC expression pattern of genes related to the differentiation status of neuroepithelium, namely SALL4, Glypican3, nestin, SOX2, Foxj1, and Ki-67. Results: Mean age of the five patients of ovarian MT with pseudostratified ependymal tubules was 26 years (range from 19 to 31 years). Two tumors were located in the left ovary and three in the right. All five cases were excised, and clinical follow-up was available (mean follow-up 1.5 years; range 0.5 to 3 years). No recurrence was noted in any cases. The pseudostratified ependymal tubules of ovarian MT, which were lined with columnar or oval epithelia up to 4-6 layers, were morphologically similar to the primitive neuroepithelial tubules of IMT and different from monolayer ependymal epithelium of ovarian MT. By immunohistochemistry, SALL4 and Glypican3 were negative, Foxj1 was positive and Ki-67 index was lower in the pseudostratified ependymal tubules and the monolayer ependymal epithelium of ovarian MT. However, the primitive neuroepithelial tubules of IMT showed variably expression of SALL4 and Glypican3, were negative for Foxj1 and high Ki-67 index. All the above three groups expressed nestin and SOX2. Conclusions: The pseudostratified ependymal tubules of ovarian MT, which have morphological similarities to the primitive neuroepithelial tubules of IMT, are similar to the monolayer ependymal epithelia of the MT in immunophenotype. IHC assessment of Foxj1 and Ki-67 is helpful to differentiate the pseudostratified ependymal tubules of ovarian MT from the primitive neuroepithelial tubules of IMT.
Female ; Humans ; Young Adult ; Adult ; Nestin ; Ki-67 Antigen ; Immunohistochemistry ; Ovarian Neoplasms/pathology* ; Teratoma/pathology*

PD-1 and PD-L1 antibodies have brought about extraordinary clinical benefits for cancer patients, and their indications are expanding incessantly. Currently, most PD-1/PD-L1 agents are administered intravenously, which may be uncomfortable for some cancer patients. Herein, we develop a novel oral-delivered small molecular, YPD-29B, which specifically targets human PD-L1. Our data suggested that YPD-29B could potently and selectively block the interaction between PD-L1 and PD-1, but did not inhibit any other immune checkpoints. Mechanistically, YPD-29B induced human PD-L1 dimerization and internalization, which subsequently activated T lymphocytes and therefore overcomes immunity tolerance in vitro. YDP-29B was modified as the YPD-30 prodrug to improve druggability. Using humanized mice with human PD-1 xenografts of human PD-L1 knock-in mouse MC38 cancer cells, we demonstrated that YPD-30 exhibited significant antitumor activity and was well tolerated in vivo. Taken together, our results indicate that YPD-30 serves as a promising therapeutic candidate for anti-human PD-L1 cancer immunotherapy.