Background::B-cell maturation antigen (BCMA)-directed chimeric antigen receptor T (CAR-T) therapy yield remarkable responses in patients with relapsed/refractory multiple myeloma (R/RMM). Circulating tumor DNA (ctDNA) reportedly exhibits distinct advantages in addressing the challenges posed by tumor heterogeneity in the distribution and genetic variations in R/RMM.Methods::Herein, the ctDNA of 108 peripheral blood plasma samples from patients with R/RMM at the First Affiliated Hospital, School of Medicine, Zhejiang University was thoroughly investigated before administration of anti-BCMA CAR-T therapy to establish its predictive potential. Flow cytometry is used primarily to detect subgroups of T cells or CAR-T cells.Results::In this study, several tumor and T cell effector-mediated factors were considered to be related to treatment failure by an integrat analysis, including higher percentages of multiple myeloma (MM) cells in the bone marrow ( P = 0.0125), lower percentages of CAR-T cells in the peripheral blood at peak ( P = 0.0375), and higher percentages of CD8 + T cells ( P = 0.0340). Furthermore, there is a substantial correlation between high ctDNA level (>143 ng/mL) and shorter progression-free survival (PFS) ( P = 0.007). Multivariate Cox regression analysis showed that high levels of ctDNA (>143 ng/mL), MM-driven high-risk mutations (including IGLL5 [ P = 0.004], IRF4 [ P = 0.024], and CREBBP [ P = 0.041]), number of multisite mutations, and resistance-related mutation ( ERBB4, P = 0.040) were independent risk factors for PFS. Conclusion::Finally, a ctDNA-based risk model was built based on the above independent risk factors, which serves as an adjunct non-invasive measure of substantial tumor burden and a prognostic genetic feature that can assist in predicting the response to anti-BCMA CAR-T therapy.

The wear debris generated during artificial joint prosthesis service can react with bone tissues to form osteolysis,seriously affecting the life-time of artificial joint prostheses.This paper reviews,summarizes,and analyzes domestic and international research literature on the extraction,characterization,and identification of wear debris from different artificial joint materials,aiming to provide references and feasible ideas for the future construction of a systematic and hierarchical research system for artificial joint wear debris.The main findings are as follows:strong alkali protein degradation test,strong acid protein degradation test,and protease protein degradation test are the commonly used method for extracting artificial joint wear debris,and researchers have clarified the protein degradation mechanisms of these three debris extraction methods.The characterization of wear debris in-vitro and in-vivo is mostly for hip and knee joints,with a small amount involving cervical spine and ankle joints.Studies have shown that the size,quantity,shape,and volume of wear particles are influenced by factors such as joint type,contact area,material selection,and implantation time.Both domestic and international studies have conducted characterization research on wear debris after in-vitro simulation testing,but there is still a lack of wear debris characterization analysis of clinical retrievals in China.Currently,most research is on the recognition of wear debris in the traditional mechanical field,but research on the intelligent recognition of artificial joint wear debris is relatively few,indicating that there is a certain lag in the application of computer technology in the field of artificial joint wear debris recognition.

The wear debris generated during artificial joint prosthesis service can react with bone tissues to form osteolysis,seriously affecting the life-time of artificial joint prostheses.This paper reviews,summarizes,and analyzes domestic and international research literature on the extraction,characterization,and identification of wear debris from different artificial joint materials,aiming to provide references and feasible ideas for the future construction of a systematic and hierarchical research system for artificial joint wear debris.The main findings are as follows:strong alkali protein degradation test,strong acid protein degradation test,and protease protein degradation test are the commonly used method for extracting artificial joint wear debris,and researchers have clarified the protein degradation mechanisms of these three debris extraction methods.The characterization of wear debris in-vitro and in-vivo is mostly for hip and knee joints,with a small amount involving cervical spine and ankle joints.Studies have shown that the size,quantity,shape,and volume of wear particles are influenced by factors such as joint type,contact area,material selection,and implantation time.Both domestic and international studies have conducted characterization research on wear debris after in-vitro simulation testing,but there is still a lack of wear debris characterization analysis of clinical retrievals in China.Currently,most research is on the recognition of wear debris in the traditional mechanical field,but research on the intelligent recognition of artificial joint wear debris is relatively few,indicating that there is a certain lag in the application of computer technology in the field of artificial joint wear debris recognition.

BACKGROUND: B-cell maturation antigen (BCMA)-directed chimeric antigen receptor T (CAR-T) therapy yield remarkable responses in patients with relapsed/refractory multiple myeloma (R/RMM). Circulating tumor DNA (ctDNA) reportedly exhibits distinct advantages in addressing the challenges posed by tumor heterogeneity in the distribution and genetic variations in R/RMM. METHODS: Herein, the ctDNA of 108 peripheral blood plasma samples from patients with R/RMM was thoroughly investigated before administration of anti-BCMA CAR-T therapy to establish its predictive potential. Flow cytometry is used primarily to detect subgroups of T cells or CAR-T cells. RESULTS: In this study, several tumor and T cell effector-mediated factors were considered to be related to treatment failure by an integrat analysis, including higher percentages of multiple myeloma (MM) cells in the bone marrow (P = 0.013), lower percentages of CAR-T cells in the peripheral blood at peak (P = 0.037), and higher percentages of CD8+ T cells (P = 0.034). Furthermore, there is a substantial correlation between high ctDNA level (>143 ng/mL) and shorter progression-free survival (PFS) (P = 0.007). Multivariate Cox regression analysis showed that high levels of ctDNA (>143 ng/mL), MM-driven high-risk mutations (including IGLL5 [P = 0.004], IRF4 [P = 0.024], and CREBBP [P = 0.041]), number of multisite mutations, and resistance-related mutation (ERBB4, P = 0.040) were independent risk factors for PFS. CONCLUSION: Finally, a ctDNA-based risk model was built based on the above independent risk factors, which serves as an adjunct non-invasive measure of substantial tumor burden and a prognostic genetic feature that can assist in predicting the response to anti-BCMA CAR-T therapy. REGISTERATION Chinese Clinical Trial Registry (ChiCTR2100046474) and National Clinical Trial (NCT04670055, NCT05430945).

Objective:To investigate the regulatory mechanism of myoepithelial cells on glandular epithelial cells during the invasive process of ductal carcinoma in situ of breast.Methods:A total of 157 patients with ductal carcinoma in situ of breast,treated at the Tianjin Medical University Cancer Hospital from May 2008 to July 2010,were randomly selected(including 63 high nuclear grade patients,51 middle nuclear grade patients,and 43 low nuclear grade patients).Immunohistochemical staining for epithelial-mesenchymal transition(EMT)-related markers(Snail and ZEB1)was performed on tumor tissue specimens from these patients to explore the correlation between tumor cell nuclear grade,EMT process,and expression status of myoepithelial cells.To further investigate the regulatory role of myoepithelial cells on glandular epithelial cells during the invasive process of ductal carcinoma in situ of breast,a co-culture model of human myoepithelial cell line Hs578Bst and adenomatous epithelial cell line MCF-7 was established using Transwell chambers.Experimental,blank control,positive control,and negative control groups were designed by combining co-cultured Hs578Bst and MCF-7 cells with exogenous TGFβ1 and TGFβ1 inhibitors.After 72 hours of culture,morphological changes,migration and proliferation capabilities of MCF-7,as well as the changes in protein and mRNA expression levels of EMT-related genes(Snail and ZEB1),were observed in each group.Results:Immunohistochemical staining results demonstrated a positive correlation between tumor nuclear atypia,EMT activation,and myoepithelial cell expression in ductal carcinoma in situ tissues.The model of ductal carcinoma in situ of breast demonstrated that myoepithelial cells Hs578Bst promoted morphological changes in glandular epithelial cells MCF-7 by stimulating TGFβ1 expression.Wound healing and cell proliferation assays revealed that myoepithelial cells Hs578Bst can enhance migration and proliferation of glandular epithelial cells MCF-7 via TGFβ1 activation.Western blot and real-time quantitative PCR confirmed that myoepithelial cells Hs578Bst can upregulate the protein and mRNA expression levels of EMT-related genes(Snail and ZEB1)in glandular epithelial cells MCF-7 by through TGFβ1 stimulation.Conclusion:Breast myoepithelial cells promote EMT in glandular epithelial cells by secreting/stimulating TGFβ1,thereby contributing to the occurrence of invasion in ductal carcinoma in situ of the breast.

In order to develop a positive quality control products for the detection of porcine repro-ductive and respiratory syndrome virus(PRRSV)nucleic acid by real-time fluorescent quantitative PCR(RT-qPCR),the positive quality control products of PRRSV-1 and PRRSV-2 M genes were prepared using armored RNA technology of MS2 phage.PRRSV-1 and PRRSV-2 M genes were amplified,purified and recovered,and ligated into pET28b vector containing MS2 mature enzyme protein gene and capsid protein.After transformed into BL21(DE3),the gene products were in-duced by IPTG and purified by PEG6000 precipitation method to prepare the armored RNA virus-like particles(AR-PRRSV)containing PRRSV M gene.Following the performance evaluation,as the positive quality control products of PRRSV-1 and PRRSV-2 M genes,AR-PRRSV1M and AR-PRRSV2M were calculated using YY/T 1652-2019 standard.Results showed that it had a good u-niformity,stable storage for the armored virus-like particles at-20,4,25 ℃ for 60 d,and 37 ℃ for 30 d.The prepared armored virus-like particles AR-PRRSV1M and AR-PRRSV2M were deter-mined by digital quantitative PCR(ddPCR)after preliminary quantification by RT-qPCR.The 104 copies/μL of AR-PRRSV1M and AR-PRRSV2M ddPCR fixation was(1.33+0.50)× 104 cop-ies/μL.The above results indicates that the AR-PRRSVM can be used as the quality control of the whole detection process(nucleic acid extraction,reverse transcription and RT-qPCR).

In response to the challenges of small scale and huge labor cost in biomedical feature modeling in current skin disease assisted diagnosis,as well as the inability to accurately describe the time series of patient disease features,a fusion text classification algorithm is used to integrate commonly used text classification models(TextLSTM,TextCNN,and RCNN)to obtain a model based on transfer learning and neural networks(TLNN model).By extracting the medical features of image sensors and quantizing them,the pretreatment reduces the number of foci and eliminates the feature information with large deviations,thus improving the accuracy of decision data.TLNN model achieves an accuracy of 72.36%on ISIC2018 and PH2 datasets,which is higher than those of the other 3 text classification models.The diagnostic accuracy of TLNN model is close to doctor's diagnosis(92%vs 94%),but the effective diagnostic efficiency is significantly higher than doctor's diagnosis(1.17 min/case vs 4.57 min/case),and the overall efficiency is improved by 290%.The results demonstrate that the fusion text classification algorithm model can obtain accurate diagnosis in less time than the traditional manual diagnosis.TLNN model can be applied to disease diagnosis,and assist doctors in medical decision-making,thereby providing patients with high-quality and convenient intelligent diagnosis and treatment services.

Objective To investigate the effect of spleen on hepatic macrophages mediated activation of hepatic stellate cells(HSCs)in mice with liver fibrosis.Methods Eighteen male C57BL/6 mice were randomly divided into three groups.Mice in Group A and Group B were injected intraperitoneally with CCl4 to establish liver fibrosis mouse model,while those in Group C were injected with corn oil as normal control.Four weeks later,mice with liver fibrosis received splenectomy(Spx)or sham operation(Sham),respectively.After continuous injection for 2 weeks,liver homogenates(L-Homo)were prepared and liver cells were isolated from the three groups.Expressions of IL-1β,IL-13,TGF-β,TNF-α,PDGF-β and VEGF in the liver homogenates of the three groups were detected by Luminex multifactor analysis.The expressions of these cytokines in liver macrophages(L-Mψ)and other non-parenchymal cells of Sham and Spx mice were analyzed by Real-time quantitative PCR(RT-qPCR)and flow cytometry.Macrophage cell line RAW264.7 or bone marrow-derived macrophages(BMDMs)were treated with liver homogenates from the Sham and Spx groups.Then the differently treated RAW264.7 cells were analyzed for mRNA expressions of cytokines and glutamine metabolism-related molecules by RT-qPCR,or transwell co-cultured with hepatic stellate cell line JS1.After co-culture,the survival and extracellular matrix expression of JS1 cells were analyzed.For comparison,Student's t test(between two groups)or one-way analysis of variance(among multiple groups)were used.Results Compared with normal control group,the concentrations of IL-1β,IL-13,TGF-β and TNF-α in the L-Homo of model group were significantly increased and showed higher levels in Sham group than in Spx group.Moreover,the hepatic macrophages were indicated as the major source of these cytokines.Consistently,macrophages treated with liver homogenate of Sham mice had increased expressions of IL-1β,TGF-β and TNF-α and glutaminase(GLS).After co-culture with macrophages treated with liver homogenate of Sham group rather than Spx group,JS1 expressed higher expressions of α-SMA and collagens.Conclusion The spleen is involved in regulating the secretion of cytokines by hepatic macrophages and enhancing their ability to activate hepatic stellate cells.

Objective : To investigate the drug resistance and pathogenicity of six clinical isolates of Klebsiella pneu- moniae (Kp) ，and to provide a basis for prevention and treatment of Kp infection． Methods : The six strains from different hospitals were isolated ，cultured ，and identified by species-specific gene khe． Their whole genome se- quences (WGS) were obtained using next-generation sequencing technology (NGS) ．Based on the WGS，the cap- sular serotypes，sequence types (ST) and drug-resistance genes of six strains were identified．The capsular sero- type genes and virulence genes were validated or identified using PCR. Broth microdilution tests were conducted to validate their drug susceptibility，and mice were challenged with Kp aerosols by MicroSprayer aerosolizer to evaluate their pathogenicity. Results : The six strains were all serotype K2 but belonged to four ST types ( ST14 ，ST65， ST700，and ST86) ，and collectively carried six virulence genes and 23 drug-resistance genes．All the six strains were resistant to ampicillin，but only one strain was multidrug-resistant．Four strains exhibited high mucoid charac- teristics．Five strains could cause mortality in mice，which were preliminary identified as high virulence strains. Conclusion For the six Kp clinical isolates from different sources，only one strain named NY 13294 is both multi- drug-resistant and highly virulent，and other four highly virulent strains are resistant to one or two types of antibiot- ics.

Objective:To investigate the regulatory mechanism of myoepithelial cells on glandular epithelial cells during the invasive process of ductal carcinoma in situ of breast.Methods:A total of 157 patients with ductal carcinoma in situ of breast,treated at the Tianjin Medical University Cancer Hospital from May 2008 to July 2010,were randomly selected(including 63 high nuclear grade patients,51 middle nuclear grade patients,and 43 low nuclear grade patients).Immunohistochemical staining for epithelial-mesenchymal transition(EMT)-related markers(Snail and ZEB1)was performed on tumor tissue specimens from these patients to explore the correlation between tumor cell nuclear grade,EMT process,and expression status of myoepithelial cells.To further investigate the regulatory role of myoepithelial cells on glandular epithelial cells during the invasive process of ductal carcinoma in situ of breast,a co-culture model of human myoepithelial cell line Hs578Bst and adenomatous epithelial cell line MCF-7 was established using Transwell chambers.Experimental,blank control,positive control,and negative control groups were designed by combining co-cultured Hs578Bst and MCF-7 cells with exogenous TGFβ1 and TGFβ1 inhibitors.After 72 hours of culture,morphological changes,migration and proliferation capabilities of MCF-7,as well as the changes in protein and mRNA expression levels of EMT-related genes(Snail and ZEB1),were observed in each group.Results:Immunohistochemical staining results demonstrated a positive correlation between tumor nuclear atypia,EMT activation,and myoepithelial cell expression in ductal carcinoma in situ tissues.The model of ductal carcinoma in situ of breast demonstrated that myoepithelial cells Hs578Bst promoted morphological changes in glandular epithelial cells MCF-7 by stimulating TGFβ1 expression.Wound healing and cell proliferation assays revealed that myoepithelial cells Hs578Bst can enhance migration and proliferation of glandular epithelial cells MCF-7 via TGFβ1 activation.Western blot and real-time quantitative PCR confirmed that myoepithelial cells Hs578Bst can upregulate the protein and mRNA expression levels of EMT-related genes(Snail and ZEB1)in glandular epithelial cells MCF-7 by through TGFβ1 stimulation.Conclusion:Breast myoepithelial cells promote EMT in glandular epithelial cells by secreting/stimulating TGFβ1,thereby contributing to the occurrence of invasion in ductal carcinoma in situ of the breast.