Objective:To investigate the application value of a joint prediction model based on lymph node imaging features in evaluating lymph node metastasis of locally advanced rectal cancer (LARC) patients after neoadjuvant chemoradiotherapy (nCRT).Methods:The retrospective cohort study was conducted. The clinicopathological data of 215 LARC patients who were admitted to Peking University Cancer Hospital & Institute from July 2010 to June 2015 were collected. There were 131 males and 84 females, aged (56.7±10.1)years. All 215 patients were randomly divided into a training set of 143 cases and a testing set of 72 cases using a 2∶1 ratio of random seed numbers. The training set was used to construct the prediction model, and the testing set was used to validate the performance of prediction model. Observation indicators: (1) lymph node metastasis in LARC patients after nCRT; (2) imaging feature selection and model construction and evaluation. Com-parison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test. Univariate and multivariate analyses were conducted using the Logistic regression model. Performance evaluation of prediction model was conducted using the receiver operating characteristic (ROC) curve. The area under the curve (AUC), accuracy, sensitivity, and specificity were calculated. Calibration curves and decision curves were used to evaluate the consistency and clinical application value of the prediction model. Results:(1) Lymph node metastasis in LARC patients after nCRT. Of the 215 LARC patients after nCRT, results of postoperative pathological examination showed that there were 162 cases with negative lymph node metastasis and 53 cases with positive lymph node metastasis, showing significant differences in age and maximum short-axis diameter of lymph node between them ( t=2.178, Z=-5.305, P<0.05). (2) Imaging feature selection and model construction and evaluation. Forty-one imaging features were extracted from the 215 LARC patients after nCRT, including 9 gray-level first-order features, 24 gray-level co-occurrence matrix features and 8 shape features. The score of lymph node (LNscore) in 162 cases with negative lymph node metastasis and 53 cases with positive lymph node metastasis were 0.18(0.10,0.33) and 0.39(0.23,0.54), respectively, showing a significant difference between them ( Z=-5.487, P<0.05). Results of multivariate analysis showed that maximum short-axis diameter of lymph node and LNscore were independent factors influencing lymph node metastasis of LARC patients after nCRT ( odds ratio=1.277, 25.514, 95% confidence interval as 1.010-1.614, 2.003-324.964, P<0.05). A Logistic regression joint prediction model was constructed by incorporating the maximum short-axis diameter of lymph node and LNscore. The ROC curves results showed that the AUC, accuracy, sensitivity, and specificity of the joint prediction model in the training set were 0.779 (95% confidence interval as 0.702-0.844), 72.7%, 71.4%, and 73.2%, respectively. The above indicators in the testing set were 0.805 (95% confidence interval as 0.694-0.889), 80.6%, 66.7%, and 85.2%, respectively. Calibration curves in both training set and test set showed good agreement with the ideal curve, indicating high calibration. Decision curves demonstrated the model′s clinical utility with a high net benefit. Conclusion:The maximum short-axis diameter of lymph node and LNscore are independent factors influencing lymph node metastasis of LARC patients after nCRT. The joint prediction model constructed based on the above indicators can be used to predict lymph node metastasis in LARC patients after nCRT.

AIM:To explore the role of stromal interaction molecule 1(STIM1)in the migration,invasion and angiogenesis of salivary adenoid cystic carcinoma(SACC),as well as its molecular mechanism.METHODS:Immu-nohistochemistry and Western blot were used to detect the expression level of STIM1 in human SACC tumor tissues and ad-jacent normal tissues.The TCGA database was analyzed to investigate the relationship between STIM1 expression and sur-vival in patients.A SACC-83 cell line with stable STIM1 overexpression(STIM1-OE)was established and divided into two groups:blank control(STIM1-Vec)group and STIM1-OE group.A nude mouse subcutaneous xenograft tumor model(n=6)was used to detect SACC growth.Transwell chamber assay,scratch test and dorsal root ganglion model were ap-plied to assess SACC migration and invasion.Immunohistochemistry was performed to detect the expression of CD34 and STIM1 in tumor tissues of nude mice.A nude mouse Matrigel plug model and human umbilical vein endothelial cell(HUVEC)tube formation assay were used to evaluate angiogenesis.Enzyme linked immunosorbent assay was employed to detect the levels of vascular endothelial growth factor(VEGF)and epidermal growth factor(EGF)in cell culture supernatants.RT-qPCR and Western blot were performed to detect the mRNA and protein expression levels of c-Myc,ribosomal protein L35(RPL35),ribosomal protein SA(RPSA),mitochondrial ribosomal protein L11(RPL11)and FAU ubiquitin like and ribo-somal protein S30 fusion(FAU).The STIM1-siRNA and RPL35-siRNA were transfected into SACC-83 cells,and the mi-gration,invasion and angiogenesis abilities of the cells were detected using the same methods as above.RESULTS:STIM1 was highly expressed in SACC,and the patients with high STIM1 expression had shorter survival time.In vivo,compared with STIM1-Vec group,STIM1-OE promoted tumor growth.In vitro,the number of migrating and invading SACC cells in STIM1-OE group was significantly increased,and the nerve invasion ability was also significantly en-hanced.Conversely,STIM1-siRNA significantly reduced the migration and invasion abilities of SACC cells.Additional-ly,STIM1-OE significantly promoted the expression of the vascular marker CD34,the secretion of VEGF and EGF,and the results of Matrigel plug and HUVEC tube formation assays indicated that STIM1-OE significantly promoted angiogene-sis.Silencing of RPL35 significantly inhibited SACC migration,invasion,and angiogenesis.RT-qPCR and Western blot results showed that the mRNA levels of RPL35,RPSA,MRPL11 and FAU were significantly increased(P<0.05),and the protein expression levels of STIM1,c-Myc and RPL35 were significantly increased(P<0.01).CONCLUSION:STIM1 drive the migration,invasion and angiogenesis of SACC by activating c-Myc/RPL35-mediated ribosome pathway.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective:To investigate the application value of a joint prediction model based on lymph node imaging features in evaluating lymph node metastasis of locally advanced rectal cancer (LARC) patients after neoadjuvant chemoradiotherapy (nCRT).Methods:The retrospective cohort study was conducted. The clinicopathological data of 215 LARC patients who were admitted to Peking University Cancer Hospital & Institute from July 2010 to June 2015 were collected. There were 131 males and 84 females, aged (56.7±10.1)years. All 215 patients were randomly divided into a training set of 143 cases and a testing set of 72 cases using a 2∶1 ratio of random seed numbers. The training set was used to construct the prediction model, and the testing set was used to validate the performance of prediction model. Observation indicators: (1) lymph node metastasis in LARC patients after nCRT; (2) imaging feature selection and model construction and evaluation. Com-parison of measurement data with normal distribution between groups was conducted using the independent sample t test. Comparison of measurement data with skewed distribution between groups was conducted using the Mann-Whitney U test. Comparison of count data between groups was conducted using the chi-square test. Univariate and multivariate analyses were conducted using the Logistic regression model. Performance evaluation of prediction model was conducted using the receiver operating characteristic (ROC) curve. The area under the curve (AUC), accuracy, sensitivity, and specificity were calculated. Calibration curves and decision curves were used to evaluate the consistency and clinical application value of the prediction model. Results:(1) Lymph node metastasis in LARC patients after nCRT. Of the 215 LARC patients after nCRT, results of postoperative pathological examination showed that there were 162 cases with negative lymph node metastasis and 53 cases with positive lymph node metastasis, showing significant differences in age and maximum short-axis diameter of lymph node between them ( t=2.178, Z=-5.305, P<0.05). (2) Imaging feature selection and model construction and evaluation. Forty-one imaging features were extracted from the 215 LARC patients after nCRT, including 9 gray-level first-order features, 24 gray-level co-occurrence matrix features and 8 shape features. The score of lymph node (LNscore) in 162 cases with negative lymph node metastasis and 53 cases with positive lymph node metastasis were 0.18(0.10,0.33) and 0.39(0.23,0.54), respectively, showing a significant difference between them ( Z=-5.487, P<0.05). Results of multivariate analysis showed that maximum short-axis diameter of lymph node and LNscore were independent factors influencing lymph node metastasis of LARC patients after nCRT ( odds ratio=1.277, 25.514, 95% confidence interval as 1.010-1.614, 2.003-324.964, P<0.05). A Logistic regression joint prediction model was constructed by incorporating the maximum short-axis diameter of lymph node and LNscore. The ROC curves results showed that the AUC, accuracy, sensitivity, and specificity of the joint prediction model in the training set were 0.779 (95% confidence interval as 0.702-0.844), 72.7%, 71.4%, and 73.2%, respectively. The above indicators in the testing set were 0.805 (95% confidence interval as 0.694-0.889), 80.6%, 66.7%, and 85.2%, respectively. Calibration curves in both training set and test set showed good agreement with the ideal curve, indicating high calibration. Decision curves demonstrated the model′s clinical utility with a high net benefit. Conclusion:The maximum short-axis diameter of lymph node and LNscore are independent factors influencing lymph node metastasis of LARC patients after nCRT. The joint prediction model constructed based on the above indicators can be used to predict lymph node metastasis in LARC patients after nCRT.

AIM:To explore the role of stromal interaction molecule 1(STIM1)in the migration,invasion and angiogenesis of salivary adenoid cystic carcinoma(SACC),as well as its molecular mechanism.METHODS:Immu-nohistochemistry and Western blot were used to detect the expression level of STIM1 in human SACC tumor tissues and ad-jacent normal tissues.The TCGA database was analyzed to investigate the relationship between STIM1 expression and sur-vival in patients.A SACC-83 cell line with stable STIM1 overexpression(STIM1-OE)was established and divided into two groups:blank control(STIM1-Vec)group and STIM1-OE group.A nude mouse subcutaneous xenograft tumor model(n=6)was used to detect SACC growth.Transwell chamber assay,scratch test and dorsal root ganglion model were ap-plied to assess SACC migration and invasion.Immunohistochemistry was performed to detect the expression of CD34 and STIM1 in tumor tissues of nude mice.A nude mouse Matrigel plug model and human umbilical vein endothelial cell(HUVEC)tube formation assay were used to evaluate angiogenesis.Enzyme linked immunosorbent assay was employed to detect the levels of vascular endothelial growth factor(VEGF)and epidermal growth factor(EGF)in cell culture supernatants.RT-qPCR and Western blot were performed to detect the mRNA and protein expression levels of c-Myc,ribosomal protein L35(RPL35),ribosomal protein SA(RPSA),mitochondrial ribosomal protein L11(RPL11)and FAU ubiquitin like and ribo-somal protein S30 fusion(FAU).The STIM1-siRNA and RPL35-siRNA were transfected into SACC-83 cells,and the mi-gration,invasion and angiogenesis abilities of the cells were detected using the same methods as above.RESULTS:STIM1 was highly expressed in SACC,and the patients with high STIM1 expression had shorter survival time.In vivo,compared with STIM1-Vec group,STIM1-OE promoted tumor growth.In vitro,the number of migrating and invading SACC cells in STIM1-OE group was significantly increased,and the nerve invasion ability was also significantly en-hanced.Conversely,STIM1-siRNA significantly reduced the migration and invasion abilities of SACC cells.Additional-ly,STIM1-OE significantly promoted the expression of the vascular marker CD34,the secretion of VEGF and EGF,and the results of Matrigel plug and HUVEC tube formation assays indicated that STIM1-OE significantly promoted angiogene-sis.Silencing of RPL35 significantly inhibited SACC migration,invasion,and angiogenesis.RT-qPCR and Western blot results showed that the mRNA levels of RPL35,RPSA,MRPL11 and FAU were significantly increased(P<0.05),and the protein expression levels of STIM1,c-Myc and RPL35 were significantly increased(P<0.01).CONCLUSION:STIM1 drive the migration,invasion and angiogenesis of SACC by activating c-Myc/RPL35-mediated ribosome pathway.

AIM:To investigate the protective effects of Klotho protein against hypoxia/reoxygenation(H/R)-in-duced damage in rat cardiomyocytes,and to elucidate its underlying mechanisms.METHODS:Rat cardiomyocyte H9c2 cells were divided into 4 groups:control,H/R,low-concentration(1 μmol/L)Klotho+H/R,and high-concentration(10 μmol/L)Klotho+H/R groups.Cells were pretreated with Klotho at specified concentrations before induction of H/R injury.Flow cytometry was used to determine cardiomyocyte apoptosis rates,while reactive oxygen species(ROS)levels were measured using the DCFH-DA probe.Additionally,superoxide dismutase(SOD)activity and malondialdehyde(MDA)content were assessed using biochemical assay kits.Mitochondrial membrane potential was evaluated using the JC-1 as-say,and activity of caspase-3 and caspase-9 was quantified.Western blot analysis was conducted to measure the protein expression of cytochrome C(Cyt-C),B-cell lymphoma-2(Bcl-2),and Bcl-2-associated X protein(Bax)in cardio-myocytes from each group.RESULTS:Compared with the control group,the H/R group exhibited significantly increased apoptosis rates(P<0.05),elevated ROS levels and MDA content,decreased SOD activity(P<0.05),reduced mitochon-drial membrane potential(P<0.05),increased caspase-3 and caspase-9 activity(P<0.05),decreased mitochondrial Cyt-C and Bcl-2 protein expression(P<0.05),and increased cytoplasmic Cyt-C and Bax protein expression(P<0.05).In comparison with the H/R group,both low-and high-concentration Klotho treatments significantly reduced cardiomyocyte apoptosis(P<0.05),lowered ROS levels and MDA content(P<0.05),increased SOD activity(P<0.05),restored mito-chondrial membrane potential(P<0.05),decreased caspase-3 and caspase-9 activity(P<0.05),increased mitochondrial Cyt-C and Bcl-2 expression(P<0.05),and decreased cytoplasmic Cyt-C and Bax expression(P<0.05).Notably,the high-concentration Klotho group demonstrated more pronounced protective effects(P<0.05).CONCLUSION:Klotho protein exerts protective effects against H/R-induced cardiomyocyte injury,possibly by inhibiting mitochondria-mediated apoptosis.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective:To investigate the prevalence of sarcopenia and potential influencing factors in middle-aged and elderly populations in Zhejiang Province.Methods:Data were obtained from Zhejiang Provincial Household Economic Status Survey, a cross-sectional survey was condcuted in middle-aged and olde adults selected through multi-stage sampling in three cities in Zhejiang (Huzhou, Jiaxing and Shaoxing) in July 2023. A total of 3 019 study participants, average age 62.3 years old, 53.5% men, were included according to the inclusion and exclusion criteria. Sarcopenia screening was conducted by using the questionnaire with five sarcopenia related-items. Univariable and multivariable logistic regression analysis was used to identify the factors associated with sarcopenia.Results:The prevalence of sarcopenia in the middle-aged and old study participants was 4.47%. Significant differences were observed between the participants with or without sarcopenia in terms of age, educational level, BMI, alcohol consumption status, diet habit, physical activity level, sleep quality, number of chronic diseases, childhood socioeconomic status, adulthood community socioeconomic status, muscle strength, walking assistance, ability to stand from seat, ability to climb stairs, and fall frequency ( P<0.05). Multivariable logistic regression analysis revealed that old age (≥75 years: OR=2.82, 95% CI: 1.60-4.97), low body weight ( OR=1.96, 95% CI: 1.06-3.62), unhealthy diet habit ( OR=1.57, 95% CI: 1.01-2.46), physical inactivity ( OR=5.80, 95% CI: 3.09-10.88), poor or very poor sleep quality ( OR=1.65, 95% CI:1.23-2.41), number of chronic diseases (1 chronic disease: OR=1.84, 95% CI: 1.08-3.14; 2 chronic diseases: OR=3.22, 95% CI: 1.81-5.71; 3 or more chronic diseases: OR=3.74, 95% CI: 2.11-6.65), poor childhood socioeconomic status ( OR=2.98, 95% CI: 1.23-7.20), and poor adulthood community socioeconomic status ( OR=3.87, 95% CI: 1.63-9.17) were significant risk factors for sarcopenia. Conclusions:The prevalence of sarcopenia was relatively low in middle-aged and old population in Zhejiang. Age, BMI, unhealthy diet, physical activity level, sleep quality, number of chronic diseases, childhood socioeconomic status, and adulthood community socioeconomic status were identified as significant influencing factors.

Sampling is the first stage of the DNA Typing in forensic science,and some studies found that the sampling technology affected the outcomes of"Contact DNA"testing.Since swabs are the most commonly used collection instrument for"Contact DNA",the researchers in forensic science field should pay more attention to study the collection efficiency of swabs.However,currently the majority of relevant research is focused on downstream testing processes,such as typing and amplification;the corresponding work on sampling performance by swabs is quite limited.Furthermore,the absence of an internationally recognized standard procedure makes it impossible to directly compare the collection results among laboratories.In this paper,a review on the use of swabs in the forensic science field is presented.It analyzes the collection efficiency,advantages and disadvantages of different swabs,and investigates the need and feasibility of developing a standardized method to evaluate the swab collection efficiency,for improving the sampling performance of swabs and ultimately increasing the detection rate of"Contact DNA"in forensic science.

MRI has been widely used in early detection,preoperative assessment,therapeutic efficacy evaluation and prognosis prediction of breast cancer due to advantages such as high soft tissue resolution,non-radiation and repeatability.The era of precision medicine has posed new challenges of MRI for breast cancer.Future research should delve into multi-sequence microscopic imaging,deeply explore the intrinsic correlation between radiomics and genomic information,furtherly improve computational power to fully exert the role of MRI for breast cancer.The advances of MRI applicated in breast cancer under the background of precision imaging were reviewed in this article.