Objective:To establish a digital quality evaluation system for Salvia miltiorrhiza-Monascus fermentation products,screen critical quality markers,and provide methodological support for their intelligent quality control.Methods:Ultra-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry(UPLC-Q-Orbitrap-MS)was employed to quantitatively analyze 34 bioactive components(e.g.,tanshinone Ⅱ A and alvianol-ic acid B)in 20 batches of fermentation products.The key markers were screened through hierarchical cluster anal-ysis and partial least squares discriminant analysis,and an intelligent discriminant model was constructed with sup-port vector machine machine learning algorithm to digitally analyze the characteristics of quality differences between batches.Results:Thirty-four common peaks were calibrated across all batches.Combined with partial least squares analysis,six key difference markers were further screened,including terpenoids such as isotanshinone Ⅱ A and tan-shinone Ⅱ A and phenolic acids such as salvianolic acid G.The support vector machine model can achieve 100％accuracy of origin discrimination by optimizing parameters jointly with genetic algorithm and grid search.Conclusion:This study developed a digital quality evaluation system for Salvia miltiorrhiza-Monascus fermentation products through a three-step analytical strategy("chemical feature exploration-marker screening-model valida-tion"),providing a transferable technical pathway for the intelligent transformation of traditional Chinese medicine quality control.

Objective:To evaluate the short-term efficacy of our modified anterior pelvic ring internal fixator(INFIX) in the treatment of anterior pelvic ring injuries.Methods:A retrospective study was conducted to analyze the clinical data from the 16 patients with pelvic anterior ring injury who had been treated with our modified INFIX at Department of Orthopaedics, The First Hospital Affiliated to Bengbu Medical University from June 2020 to June 2023. There were 9 males and 7 females with an age of (49.1±14.3) years. According to the AO/OTA classification, 10 cases were of type B2 and 6 cases of type C1. Their time from injury to surgery was (7.6±2.9) days. Fixation with our modified INFIX was as follows: Three pedicle screws were inserted into the anterior inferior iliac spine on one side and into the pubic symphysis on both sides. Next, a subcutaneous tunnel was created from the anterior inferior iliac spine incision toward the pubic symphysis, and connecting rods were inserted for fixation. The surgical incision length, intraoperative blood loss, surgical time, postoperative fracture reduction quality, fracture healing time, incidence of complications during follow-up, and pelvic functional recovery at the last follow-up were recorded in this cohort.Results:In this cohort, surgical incision length was (5.8±0.4) cm, intraoperative blood loss (75.4±11.9) mL, and surgical time (66.1±8.9) min. By the end of one week after surgery, the quality of fracture reduction was evaluated according to the Matta scoring criteria as excellent in 11 cases and as good in 5 cases. All patients were followed up for (17.4±3.1) months after surgery. The fractures got united in all the 16 patients after (11.2±1.2) weeks. At the last follow-up, the pelvic function recovery was evaluated according to the Majeed scoring system as excellent in 13 cases and as good in 3 cases, yielding a Majeed score of (90.1±4.2) points. During the follow-up period, no patient developed complications such as nerve injury, wound infection, or loosening, breakage or withdrawal of internal fixation devices.Conclusion:In the treatment of anterior pelvic ring injuries, our modified INFIX has the advantages of few complications, simple operation and minimal invasion, leading to good short-term efficacy.

The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P＜0.05)and IFN-λ(P＜0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.

Objective:To investigate the regulatory role of the tumor suppressor gene ARID1A(AT-rich interaction domain 1A)in the proliferation of lung adenocarcinoma(LUAD)cells and its molecular mechanism associated with the protein kinase B(AKT)signaling pathway.Methods:Stable ARID1A-overexpressing A549/H1299 cell models were constructed via lentiviral transfection,with transfection efficiency validated by RT-PCR and Western blot.Cell viability was assessed using the CCK-8 assay,proliferation rate was evaluated using EdU staining,and migration capacity was analyzed by scratch assay.Cell death was evaluated through Calcein/PI live/dead staining and trypan blue exclusion.The phosphorylation level of AKT(p-AKT/AKT ratio)was detected by Western blot.The role of AKT in proliferation regulation was further validated by treating overexpressing cells with the AKT inhibitor MK-2206(1 μmol/L).Results:ARID1A overexpression significantly inhibited the proliferation and migration of A549/H1299 cells while upregulating p-AKT levels.MK-2206 treatment abolished the inhibitory effects of ARID1A overexpression on proliferation.Cell death assays demonstrated no significant impact of ARID1A overexpression on LUAD cell mortality.Conclusion:ARID1A specifically suppresses LUAD cell proliferation and migration by activating the AKT signaling pathway,suggesting that targeting AKT may provide a potential therapeutic strategy for LUAD patients with ARID1A deficiency.

Purpose To explore the clinicopathological features and potential clinical value of marginal zone lym-phoma(MZL)derived from T-bet positive memory B cell.Methods Clinical data of 67 cases of MZL were collected.Hematoxylin-eosin,immunohistochemistry,and multiple immunofluorescence stains,B cell receptor high throughput sequencing technology were used to study the histology,immunophenotype,and immunoglobulin heavy chain variable gene(IGHV)repertoire.Results T-bet was expressed in some MZL patients(34/67,50.75％),which was correla-ted with clinicopathological characteristics such as gender,clinical stage,and Ki67 proliferation index(P＜0.05),and also the progression-free survival was poor(P=0.012 2).T-bet positivity was a risk factor affecting the progres-sion of MZL.Microscopically,T-bet positive MZL frequently presented T-bet positive tumor B cells surrounded follicu-lar germinal center,"MALT ball"-type lymphoepithelial lesions,and IgG positive neoplastic plasma cells(P＜0.05).T-bet had no biased influence on the VH gene usage(P＞0.05).The common VH families were IGHV4 and IGHV3,and the segments were IGHV4-34 and IGHV3-30.The positivity of T-bet was associated with somatic hypermutation(SHM)state(P=0.014 9).The SHM was mainly in the range of 2％-4.9％in T-bet positive MZL,while in T-bet negative MZL the SHM was mostly greater than 5％.The VH gene usage was not correlated with the clinicopathological features of patients(P＞0.05).IGHV4 was correlated with progression-free survival in T-bet positive MZL(P=0.038 2).Conclusion The expression of T-bet in MZL is closely related to the clinicopathological features such as histology,plasma cell immunophenotype and IGHV gene repertoire,and the prognosis of patients is poor,which may be a potential molecular marker affecting the progression of MZL.

Tumors exhibit abnormal glucose metabolism, consuming excessive glucose and excreting lactate, which constructs a tumor microenvironment that facilitates cancer progression and disrupts immunotherapeutic efficacy. Currently, tumor glucose metabolic dysregulation to reshape the immunosuppressive microenvironment and enhance immunotherapy efficacy is emerging as an innovative therapeutic strategy. However, glucose metabolism modulators lack specificity and still face significant challenges in overcoming tumor delivery barriers, microenvironmental complexity, and metabolic heterogeneity, resulting in poor clinical benefit. Nanomedicines, with their ability to selectively target tumors or immune cells, respond to the tumor microenvironment, co-deliver multiple drugs, and facilitate combinatorial therapies, hold significant promise for enhancing immunotherapy through tumor glucose metabolic reprogramming. This review explores the complex interactions between tumor glucose metabolism-specifically metabolite transport, glycolysis processes, and lactate-and the immune microenvironment. We summarize how nanomedicine-mediated reprogramming of tumor glucose metabolism can enhance immunotherapy efficacy and outline the prospects and challenges in this field.

BACKGROUND:Gradient artificial bone repair scaffolds can mimic unique anatomical features in musculoskeletal tissues,showing great potential for repairing injured musculoskeletal tissues. OBJECTIVE:To review the latest research advances in gradient artificial bone repair scaffolds for tissue engineering in the musculoskeletal system and describe their advantages and fabrication strategies. METHODS:The first author of the article searched the Web of Science and PubMed databases for articles published from 2000 to 2023 with search terms"gradient,bone regeneration,scaffold".Finally,76 papers were analyzed and summarized after the screening. RESULTS AND CONCLUSION:(1)As an important means of efficient and high-quality repair of skeletal system tissues,gradient artificial bone repair scaffolds are currently designed bionically for the natural gradient characteristics of bone tissue,bone-cartilage,and tendon-bone tissue.These scaffolds can mimic the extracellular matrix of native tissues to a certain extent in terms of structure and composition,thus promoting cell adhesion,migration,proliferation,differentiation,and regenerative recovery of damaged tissues to their native state.(2)Advanced manufacturing technology provides more possibilities for gradient artificial bone repair scaffold preparation:Gradient electrospun fiber scaffolds constructed by spatially differentiated fiber arrangement and loading of biologically active substances have been developed;gradient 3D printed scaffolds fabricated by layered stacking,graded porosity,and bio-3D printing technology;gradient hydrogel scaffolds fabricated by in-situ layered injections,simple layer-by-layer stacking,and freeze-drying method;and in addition,there are also scaffolds made by other modalities or multi-method coupling.These scaffolds have demonstrated good biocompatibility in vitro experiments,were able to accelerate tissue regeneration in small animal tests,and were observed to have significantly improved histological structure.(3)The currently developed gradient artificial bone repair scaffolds have problems such as mismatch of gradient scales,unclear material-tissue interactions,and side effects caused by degradation products,which need to be further optimized by combining the strengths of related disciplines and clinical needs in the future.

Objective:To investigate the regulatory role of the tumor suppressor gene ARID1A(AT-rich interaction domain 1A)in the proliferation of lung adenocarcinoma(LUAD)cells and its molecular mechanism associated with the protein kinase B(AKT)signaling pathway.Methods:Stable ARID1A-overexpressing A549/H1299 cell models were constructed via lentiviral transfection,with transfection efficiency validated by RT-PCR and Western blot.Cell viability was assessed using the CCK-8 assay,proliferation rate was evaluated using EdU staining,and migration capacity was analyzed by scratch assay.Cell death was evaluated through Calcein/PI live/dead staining and trypan blue exclusion.The phosphorylation level of AKT(p-AKT/AKT ratio)was detected by Western blot.The role of AKT in proliferation regulation was further validated by treating overexpressing cells with the AKT inhibitor MK-2206(1 μmol/L).Results:ARID1A overexpression significantly inhibited the proliferation and migration of A549/H1299 cells while upregulating p-AKT levels.MK-2206 treatment abolished the inhibitory effects of ARID1A overexpression on proliferation.Cell death assays demonstrated no significant impact of ARID1A overexpression on LUAD cell mortality.Conclusion:ARID1A specifically suppresses LUAD cell proliferation and migration by activating the AKT signaling pathway,suggesting that targeting AKT may provide a potential therapeutic strategy for LUAD patients with ARID1A deficiency.

Objective:To investigate the characteristics of visual and auditory sustained attention and sex differences in pupils of different grades.Method:Totally 231 students from grades 1 to 6 participated in this stud-y.The development and sex differences of sustained attention were measured by average reaction times,coefficients of variation for reaction times,omission error rates and commission error rates in visual and auditory two-choice re-action time(CRT)tasks.Result:The students in grade 5 had significantly slower visual average reaction times and coefficients of variation for reaction times(P＜0.001 or P＜0.05)than those in grade 3.The students in grade 5 had significantly lower auditory average reaction times,coefficients of variation for reaction times,omission error rates and commission error rates(P＜0.001 or P＜0.01)than those in grade 1.There was no significant sex differ-ences in visual or auditory average reaction times,coefficients of variation for reaction times or omission error rates(Ps＞0.05).Conclusion:There would be age differences in characteristics of both visual and auditory sustained at-tention in pupils,and may be no sex difference in visual or auditory sustained attention.

Objective To investigate the safety and validity of laparoscopic autologous lingual mucosal graft ureteroplasty for the treatment of complex ureteral stricture.Methods A total of 10 patients who underwent laparoscopic autologous lingual mucosal graft ureteroplasty in our hospital from May 2021 to October 2023 were retrospectively analyzed.During the operation,the narrow segment was longitudinally dissected,and according to the length of stricture,the lingual mucosal graft of 2.0-7.0 cm in length and 1.0-1.5 cm in width was harvested and precisely anastomosed with the stenosed ureter,followed by double J stent placement.Results All the operations were successfully completed with no conversion to open surgery or intraoperative complications.The operative duration was(237.0±67.1)min,the estimated blood loss was 25.0(20.0,30.0)ml,the duration of drainage tube indwelling was 4.0(4.0,4.8)d,the duration of urinary catheter indwelling was 6.5(6.0,9.5)d,and the duration of postoperative hospitalization was 6.0(6.0,6.8)d.All the patients'oral function recovered well within 1 week,and the double J stent was removed 1-2 months after the surgery.The mean follow-up time was(12.3±7.1)months.One case of aggravated hydronephrosis on the affected side underwent a second laparoscopic ureteral stricture resection and end-to-end anastomosis.The remaining 9 cases showed significant improvement in hydronephrosis on the affected side,with improved renal pelvis separation[(2.9±1.2)cm,t=8.022,P=0.000]and renal function compared to before surgery.Their blood creatinine was(74.3±25.5)μmol/L,with no significant difference compared to preoperation[(80.1±26.6)μmol/L,t=1.825,P=0.105].Conclusion Laparoscopic autologous lingual mucosal graft ureteroplasty for the treatment of complex ureteral stricture is a safe and feasible ureteral reconstruction technique with advantages of quick recovery and reliable outcomes.